Termination and initial branch formation of SNAP-25-deficient thalamocortical fibres in heterochronic organotypic co-cultures.
We are interested in the role of neural activity mediated through regulated vesicular release in the stopping and early branching of the thalamic projections in the cortex. Axon outgrowth, arrival at the cortical subplate, side-branch formation during the waiting period and cortical plate innervation of embryonic thalamocortical projections occurs without major abnormalities in the absence of regulated release in Snap25 (-/-) null mutant mice [Washbourne et al. (2002) Nat. Neurosci. 5:19-26; Molnár et al. (2002) J. Neurosci. 22:10313-10323]. The fact that Snap25 (-/-) null mutant mice die at birth limited our previous experiments to the prenatal period. We therefore investigated the behaviour of thalamic projections in co-culture paradigms by using heterochronic thalamic [embryonic day (E)16-E18] and cortical [postnatal day (P)0-P3] explants, in which the stopping and branching behaviour has been previously documented. Our current co-culture experiments established that thalamic projections from E16-E18 Snap25(+/+) or Snap25 (-/-) explants behaved in an identical fashion in P0-P3 Snap25 (+/+) cortical explants after 7 days in vitro. Thalamic projections from Snap25 (-/-) explants developed similar patterns of fibre ingrowth to the cortex, and stopped and formed branches at a similar depth in the Snap25(+/+) cortical slice as in control cultures. These results imply that thalamic projections can reach their ultimate target cells in layer 4, stop, and start to develop branches in the absence of regulated vesicular transmitter release from their own terminals.


Scarcity of mutations detected in families with X linked hypohidrotic ectodermal dysplasia: diagnostic implications.
Indirect molecular diagnosis of X linked hypohidrotic ectodermal dysplasia (XLHED), a congenital disorder of hair, teeth, and eccrine sweat glands, has been possible by linkage analysis. Direct mutation detection would enable carrier detection in female relatives of sporadic cases, as well as help distinguish XLHED from the rarer, clinically indistinguishable, autosomal recessive disorder ARHED. Recently, a candidate gene for XLHED has been identified. Genomic DNA from 162 affected males and 21 females, who were either obligate carriers or had manifestations of the disorder, were screened by SSCP analysis. A subset of the patients had been previously screened for large genomic deletions and had limited screening of a single exon by SSCP analysis. The two known exons were amplified using flanking primers. Approximately 7% of patients, all males, had putative mutations identified within exon 1, but no variants were found within exon 2. Ten different putative mutations and four probable polymorphisms were identified. Both of the known exons were sequenced in 10 patients who had no detectable SSCP changes, but no additional mutations were found. No correlation between phenotype and genotype was evident between either affected subjects or subjects with or without detectable mutations. The results of the study indicate that only a small minority of affected males can be diagnosed by direct mutation analysis, and that the remainder of the patients are likely to have mutations in as yet unidentified exons of the EDA gene. Linkage analysis, in informative situations, therefore remains the only practical diagnostic option available.


Use of spoligotyping to study the evolution of the direct repeat locus by IS6110 transposition in Mycobacterium tuberculosis.
Based on the variability of 43 spacers within the direct repeat (DR) locus of Mycobacterium tuberculosis complex organisms, spoligotyping is a rapid method that aids in the study of the epidemiology of tuberculosis. It was recently hypothesized that despite its presence in the DR locus, spacer 31 could not be amplified in M. tuberculosis clinical isolates belonging to spoligotype 50 due to the insertion of an extra copy of IS6110 between spacers 31 and 32 that could lead to an asymmetrical split of the primer targets (I. Filliol, C. Sola, and N. Rastogi, J. Clin. Microbiol. 38:1231--1234, 2000). In the present investigation, previous observations were extended to 25 clinical isolates of type 50 showing that the primer set IS6-DRb that selectively amplified the left and central DR regions was indeed able to demonstrate the presence of spacer 31. IS6110-restriction fragment length polymorphism (RFLP) and DR-RFLP showed that type 50 isolates were characterized by the presence of two copies of IS6110 associated with the DR locus and an additional double IS6110 band of 1.4 kb. The primer set IS3-IS6 was then used to selectively amplify a 750-bp inter-IS6110 fragment within the DR locus. The sequencing of the central DR region corroborated our previous findings and showed that the absence of spacer 31 among the type 50 isolates was due to the asymmetric insertion of an extra copy of IS6110 between spacers 31 and 32, leading to an unequal split of the DRa-DRb target into two portions, of 6 and 30 bp, respectively. These results show that the DR locus constitutes an ideal IS6110 preferential locus (ipl), permitting the insertion of two or more copies of IS6110, and provide new clues for epidemiological and phylogenetic interpretation of changes in IS6110-RFLP and spoligotyping profiles.


Alkaloid defenses of co-mimics in a putative Müllerian mimetic radiation.
Polytypism in aposematic species is unlikely according to theory, but commonly seen in nature. Ranitomeya imitator is a poison frog species exhibiting polytypic mimicry of three congeneric model species (R. fantastica, R. summersi, and two morphs of R. variabilis) across four allopatric populations (a "mimetic radiation"). In order to investigate chemical defenses in this system, a key prediction of Müllerian mimicry, we analyzed the alkaloids of both models and mimics from four allopatric populations. In this study we demonstrate distinct differences in alkaloid profiles between co-mimetic species within allopatric populations. We further demonstrate that R. imitator has a greater number of distinct alkaloid types than the model species and more total alkaloids in all but one population. Given that R. imitator is the more abundant species in these populations, R. imitator is likely driving the majority of predator-learned avoidance in these complexes. The success of Ranitomeya imitator as a putative advergent mimic may be a direct result of differences in alkaloid sequestration. Furthermore, we propose that automimicry within co-mimetic species is an important avenue of research.


Miniglucagon (glucagon 19-29), a potent and efficient inhibitor of secretagogue-induced insulin release through a Ca2+ pathway.
Using the MIN6 B-cell line, we investigated the hypothesis that miniglucagon, the C-terminal () fragment processed from glucagon and present in pancreatic A cells, modulates insulin release, and we analyzed its cellular mode of action. We show that, at concentrations ranging from 0.01 to 1000 pM, miniglucagon dose-dependently (ID50 = 1 pM) inhibited by 80-100% the insulin release triggered by glucose, glucagon, glucagon-like peptide-1-(7-36) amide (tGLP-1), or glibenclamide, but not that induced by carbachol. Miniglucagon had no significant effects on cellular cAMP levels. The increase in 45Ca2+ uptake induced by depolarizing agents (glucose or extracellular K+), by glucagon, or by the Ca2+channel agonist Bay K-8644 was blocked by miniglucagon at the doses active on insulin release. Electrophysiological experiments indicated that miniglucagon induces membrane hyperpolarization, probably by opening potassium channels, which terminated glucose-induced electrical activity. Pretreatment with pertussis toxin abolished the effects of miniglucagon on insulin release. It is concluded that miniglucagon is a highly potent and efficient inhibitor of insulin release by closing, via hyperpolarization, voltage-dependent Ca2+ channels linked to a pathway involving a pertussis toxin-sensitive G protein.


In vitro testing of cellular response to ultra high frequency electromagnetic field radiation.
The aim of this study was to evaluate whether low-level, ultra high frequency (UHF) irradiation of 935 MHz influences the cell structure and growth of V79 cells. UHF field was generated inside a Gigahertz Transversal Electromagnetic Mode cell (GTEM-cell) with a Hewlett-Packard signal generator. The electric field strength was 8.2+/-0.3 V/cm and the average specific absorption rate (SAR) was calculated to be 0.12 W/kg. Cell samples were cultivated in a humidified atmosphere at 37 degrees C with 5% CO2. Prepared cell samples were exposed to a 935 MHz continuous wave frequency field for 1, 2, and 3 h. The structure of microtubule proteins has been determined using the immunocytochemical method. Cell growth was determined by cell counts for each hour of exposure during five post-exposure days. Negative- and positive-cell controls were included into the experimental procedure. In comparison with control cells, the microtubule structure clearly altered after 3h of irradiation (p<0.05). Significantly decreased growth was noted in cells exposed for 3h three days after irradiation (p<0.05). It seems that the 935 MHz, low-level UHF radiation affects microtubule proteins, which consequently may obstruct cell growth.


Cerebral infarction in Alzheimer's disease is associated with severe amyloid angiopathy and hypertension.
To determine if severe cerebral amyloid angiopathy (AA) in patients with Alzheimer's disease (AD) is associated with an increased prevalence of cerebral infarction diagnosed at autopsy. Amyloid angiopathy is increasingly recognized as a cause of ischemic infarcts, as well as cerebral hemorrhages. However, the relationship of AA to cerebral infarction in patients with AD is uncertain. Retrospective clinicopathological study of autopsy-confirmed cases of AD. One hundred forty-five deceased patients with AD confirmed at autopsy. Semiquantitative scores of AA severity were done in four brain regions: midfrontal, inferior parietal, superior temporal, and hippocampal. The finding of cerebral infarction at autopsy was modeled as a function of AA severity, hypertension, age at death, AD severity, and sex in chi 2 and multiple logistic regression analyses. Severe AA was significantly associated with cerebral infarction at autopsy in patients with AD (odds ratio [OR], 3.5; 95% confidence interval [CI], 1.4 to 8.9). None of the other independent variables in the multiple logistic regression analysis were significant predictors. While hypertension was equally common in the severe and mild AA subgroups, the combination of both severe AA and hypertension interacted to increase the risk of infarction (OR, 14.2; 95% CI, 3.2 to 63.4) beyond that observed with hypertension (OR, 1.1; 95% CI, 0.4 to 3.2) or severe AA (OR, 1.3; 95% CI, 0.3 to 5.3) alone. Severe AA is associated with an increased frequency of cerebral infarction in patients with AD. This appears to be largely due to an interaction between severe AA and hypertension that may produce multiplicative injuries on the vasculature. Further study with regard as to how AA may cause ischemia and its role in the neuropathologic and clinical progression of AD is needed.


The ultrastructure of ameloblasts during matrix formation and the maturation of enamel.
Ameloblasts from different regions of upper incisors of rats were examined with the electron microscope. During matrix formation, the cells resemble secretory cells. They are extremely long, tightly packed, and show considerable polarity. Nuclei are at the basal end of the cell. Mitochondria are proximal and the Golgi apparatus distal to the nucleus. Ergastoplasm is found in all levels but mainly in the distal end. A terminal bar apparatus separates the distal end of the cell from Tomes's process. Next to this is soft enamel. The next incisal region is a transitional zone in which the ameloblasts separate easily from the enamel. Endoplasmic reticulum is dilated and very obviously in communication with the perinuclear space. Mitochondria are present not only proximal, but also distal, to the nucleus. The next incisal zone consists of cells related to the maturation of enamel. They no longer resemble secretory cells, but now have more characteristics of transporting cells. Processes from the distal end of the cell are present with mitochondria closely applied to the base of the processes. A considerable amount of intercellular space exists with microvilli projecting into the space. Iron granules appear in these cells, and the ergastoplasmic cisternae are dilated. In the incisal end of this zone, the iron granules form aggregates. The iron finally leaves the cells to enter the enamel. Free RNP particles and fibrils become more evident after the iron leaves the cells. In the most incisal region, the ameloblasts are further reduced in height. Distal processes are no longer present and fibrils are more conspicuous.


Use of lipid-based nutrient supplements (LNS) to improve the nutrient adequacy of general food distribution rations for vulnerable sub-groups in emergency settings.
The term 'lipid-based nutrient supplements' (LNS) refers generically to a range of fortified, lipid-based products, including products like Ready-to-Use Therapeutic Foods (RUTF) (a large daily ration with relatively low micronutrient concentration) as well as highly concentrated supplements (1-4 teaspoons/day, providing <100 kcal/day) to be used for 'point-of-use' fortification. RUTF have been successfully used for the management of severe acute malnutrition (SAM) among children in emergency settings. Recent research on smaller doses of LNS for prevention of malnutrition has created interest in their potential use in emergency settings to ensure a more nutritionally adequate ration for the most vulnerable groups [e.g. infants and children between 6 and 24 months of age, and pregnant and lactating women (PLW)]. Currently, the main food and nutrition interventions in emergency settings include general food distribution (GFD) rations, which are provided to the affected population as a whole, and selective (or supplementary) feeding programs (SFP), which are to be provided to nutritionally vulnerable or malnourished individuals. In addition to logistical and operational challenges that may limit the intended effect of these programs, the nutritional quality of the food commodities provided may be insufficient to meet the needs of infants and young children and PLW. Because these subgroups have particularly high nutrient needs for growth and development, meeting these needs is challenging in settings where the ration is limited to a few food commodities, with little access to a diverse diet and bioavailable sources of micronutrients. In recent years, there has been increased attention to adding micronutrient interventions, on top of the other food-based interventions (such as GFDs and SFPs), to fill micronutrient gaps in diets in emergency settings. The focus of this document is the potential role of LNS in meeting the nutritional needs of these vulnerable subgroups, with the goal of preventing malnutrition in emergency-affected populations. The document addresses the desired nutritional formulation of LNS for these target groups, taking into account the expected bioavailability of relevant nutrients and toxicity concerns. It also discusses the recommended chemical forms of the fortificants in LNS; stability and shelf-life considerations; production, packaging and distribution of LNS in the context of emergencies; and cost implications of the addition of LNS to current GFD rations for vulnerable groups. To develop the desired nutritional formulation of LNS for these purposes, we calculated the current nutrient content of commonly provided GFD rations and determined the nutritional 'gaps' (of both micro- and macronutrients) of these rations for each of the target groups (i.e. children 6-35 months of age and PLW). For fat and protein, both quantity and quality were evaluated. Through an iterative process, we determined the formulation of a small dose of LNS that would best meet the recommended nutrient intakes for each group in combination with other foods in the GFD ration [composed of a grain, pulse, oil, sugar and salt, but excluding a fortified blended food (FBF)], as well as breast milk for children 6-24 months of age, while avoiding excess levels of any one nutrient to the extent possible. The composition of the LNS used for these calculations is based on an existing LNS product (Nutributter, Malaunay, France, Nutriset), but with less sugar and more oil. Two different approaches were used: (1) developing two different formulations of LNS, one to be used for infants and children 6-35 months of age and a separate one for PLW; and (2) developing a single formulation that could be used for all of these subgroups. We used commodity cost data to estimate the cost of adding an LNS product to the GFD ration. The results indicate that the typical GFD ration currently provided in emergency settings--based on cereals, pulse, an FBF such as corn-soy blend (CSB), oil, salt and sugar-does not meet the nutritional needs of infants and young children and PLW. The hypothetical intake from a ration composed of food aid commodities (based on the current USAID/USDA specifications for exported food aid commodities used in emergency settings), and including breast milk for children 6-24 months of age, provided less than 75% of the recommended intake for several micronutrients for certain age/physiologic groups, including calcium, iron, zinc, B vitamins such as riboflavin, B6 and B12, and fat-soluble vitamins such as D, E and K. It also generally contained lower than recommended levels of fat and essential fatty acids. The initial LNS formulation for each target group was designed to provide 100% of the recommended amount (RDA or RNI) for most micronutrients per daily dose (20 g, approximately 118 kcal) of LNS. This would ensure consumption of the recommended levels of each nutrient even if the 'base' diet changed. However, because such a formulation could provide excess amounts of certain nutrients when consumed in combination with the 'base' diet (especially when the 'base' diet contains fortified foods), we made adjustments in the LNS formulation when there was a risk of greatly exceeding the Upper Level for certain subgroups and there were relevant concerns about adverse effects from chronic consumption of such amounts. For most nutrients, consumption of toxic amounts is highly unlikely with the proposed LNS formulations. The 'one-size' LNS formulation was designed so that one 'dose' (20 g) would be provided to infants and young children and two 'doses' (i.e. 40 g/day) would be provided to PLW. This 'one-size' formulation was based on the LNS formulation developed for children 6-35 months of age. Although the resulting formulation is not a perfect match for the unique nutritional needs of each subgroup, there are several practical advantages to using such an approach. As anticipated, addition of LNS to the GFD ration, even after eliminating the FBF (e.g. CSB), increases the cost. The 'revised' ration without CSB but with LNS would cost 34-52% more (food only) than the 'typical' GFD diet for a hypothetical mother-infant pair, depending on how many LNS 'doses' were provided to the mother. However, depending on the contribution of food costs to overall program costs, the overall increase in costs may be significantly less. Although cost is an important consideration, options to improve the nutritional quality of foods provided in emergency settings should also be assessed with regard to effectiveness in maintaining and improving nutritional outcomes. Another consideration is whether a specialized product like LNS is more easily targeted to the individuals for whom it is intended, thus reducing inter- and intra-household sharing, a common concern with other fortified products such as CSB. This could have substantial cost implications because programs usually compensate for sharing by inflating the amount of FBF provided. This document is intended to be a starting point for considering the incorporation of LNS in the food packages provided in emergency settings. Our goal was to examine the potential nutritional benefits but also the challenges of adopting such a strategy. There are many different options for emergency nutrition programs, and there are also many considerations governing which option to choose. This document is intended to encourage further evaluation of all of these options.


Tissue engineering of oral mucosa: a shared concept with skin.
Tissue-engineered oral mucosa, in the form of epithelial cell sheets or full-thickness oral mucosa equivalents, is a potential solution for many patients with congenital defects or with tissue loss due to diseases or tumor excision following a craniofacial cancer diagnosis. In the laboratory, it further serves as an in vitro model, alternative to in vivo testing of oral care products, and provides insight into the behavior of the oral mucosal cells in healthy and pathological tissues. This review covers the old and new generation scaffold types and materials used in oral mucosa engineering; discusses similarities and differences between oral mucosa and skin, the methods developed to reconstruct oral mucosal defects; and ends with future perspectives on oral mucosa engineering.


Absence of carboxypeptidase E leads to adult hippocampal neuronal degeneration and memory deficits.
Molecules that govern the formation, integrity, and function of the hippocampus remain an important area of investigation. Here we show that absence of the proneuropeptide processing enzyme, carboxypeptidase E (CPE) in CPE knock-out (KO) mice had a profound effect on memory, synaptic physiology, and the cytoarchitecture of the hippocampus in these animals. Adult CPE-KO mice displayed deficits in memory consolidation as revealed by the water-maze, object preference, and social transmission of food preference tests. These mice also showed no evoked long-term potentiation. Additionally, CPE-KO mice at 4 weeks of age and older, but not at 3 weeks of age, exhibited marked degeneration specifically of the pyramidal neurons in the hippocampal CA3 region which normally expresses high levels of CPE. Immunohistochemistry revealed that the neuronal marker, NeuN, was reduced, while the glial marker, GFAP, was increased, characteristic of gliosis in the CA3 area of CPE-KO mice. Calbindin staining indicated early termination of the mossy fibers before reaching the CA1 region in these mice. Thus, absence of CPE leads to degeneration of the CA3 neurons and perturbation of the cytoarchitecture of the hippocampus. Ex vivo studies showed that overexpression of CPE in cultured hippocampal neurons protected them against H(2)O(2) oxidative-stress induced cell death. These findings taken together indicate that CPE is essential for the survival of adult hippocampal CA3 neurons to maintain normal cognitive function.


Renal medullary interstitial infusion of diltiazem alters sodium and water excretion in rats.
The role of renal papillary blood flow in regulation of fluid and electrolyte excretion was examined. The effects of an acute infusion of diltiazem (5 micrograms.kg-1 x min-1) into the renal medullary interstitium on papillary blood flow and sodium and water excretion were studied. Changes of renal blood flow were measured using an electromagnetic flow probe. Cortical and papillary blood flows were measured using laser-Doppler flowmetry. Renal and cortical blood flows were unchanged during medullary interstitial infusion of diltiazem, but papillary blood flow increased 26% (P < 0.05) and remained elevated for 1 h after diltiazem infusion was discontinued. Glomerular filtration rate (GFR) of the infused kidney increased by 21% from a control of 1.0 +/- 0.1 ml.min-1 x g-1 during infusion of diltiazem (P < 0.05), but it returned to control after diltiazem infusion was stopped. Urine flow and sodium excretion increased by 70% (P < 0.05), and fractional sodium excretion rose from 1.5 +/- 0.2 to 2.4 +/- 0.3% of the filtered load during the hour after diltiazem infusion. Renal blood flow, cortical and papillary blood flow, GFR, urine flow, and sodium excretion in the 0.9% sodium chloride vehicle-infused kidney were not significantly altered during the experiment. Intravenous infusion of the same dose of diltiazem (5 micrograms.kg-1 x min-1) increased GFR by 22%, but had no effect on urine flow and sodium excretion. These results indicate that renal medullary interstitial infusion of diltiazem selectively increased renal papillary blood flow, which was associated with an increase of sodium and water excretion.


P2X7R-Panx1 Complex Impairs Bone Mechanosignaling under High Glucose Levels Associated with Type-1 Diabetes.
Type 1 diabetes (T1D) causes a range of skeletal problems, including reduced bone density and increased risk for bone fractures. However, mechanisms underlying skeletal complications in diabetes are still not well understood. We hypothesize that high glucose levels in T1D alters expression and function of purinergic receptors (P2Rs) and pannexin 1 (Panx1) channels, and thereby impairs ATP signaling that is essential for proper bone response to mechanical loading and maintenance of skeletal integrity. We first established a key role for P2X7 receptor-Panx1 in osteocyte mechanosignaling by showing that these proteins are co-expressed to provide a major pathway for flow-induced ATP release. To simulate in vitro the glucose levels to which bone cells are exposed in healthy vs. diabetic bones, we cultured osteoblast and osteocyte cell lines for 10 days in medium containing 5.5 or 25 mM glucose. High glucose effects on expression and function of P2Rs and Panx1 channels were determined by Western Blot analysis, quantification of Ca2+ responses to P2R agonists and oscillatory fluid shear stress (± 10 dyne/cm(2)), and measurement of flow-induced ATP release. Diabetic C57BL/6J-Ins2Akita mice were used to evaluate in vivo effects of high glucose on P2R and Panx1. Western blotting indicated altered P2X7R, P2Y2R and P2Y4R expression in high glucose exposed bone cells, and in diabetic bone tissue. Moreover, high glucose blunted normal P2R- and flow-induced Ca2+ signaling and ATP release from osteocytes. These findings indicate that T1D impairs load-induced ATP signaling in osteocytes and affects osteoblast function, which are essential for maintaining bone health.


Is the issue the price of a child's life, or the futility of heroic measures?
Britain's High Court has been asked to rule whether a $160,000 experimental treatment with a 1% chance of saving the life of a 10-year-old girl should be paid for by the National Health Service. Child B, who has had one bone-marrow transplant, has had acute myeloblastic leukemia for a year and is near death. However, after her Cambridge doctors advised that further treatment would not be beneficial, her family found a hematologist willing to try an experimental treatment and a second transplant. The media debate in Britain has focused on financial aspects of the situation, not on the usually futile use of heroic measures.


System Life Cycle Evaluation(SM) (SLiCE): harmonizing water treatment systems with implementers' needs.
One of the methods proposed to improve access to clean drinking water is the mobile packaged water treatment system (MPWTS). The lack of published system performance comparisons combined with the diversity of technology available and intended operating conditions make it difficult for stakeholders to choose the system best suited for their application. MPWTS are often deployed in emergency situations, making selection of the appropriate system crucial to avoiding wasted resources and loss of life. Measurable critical-to-quality characteristics (CTQs) and a system selection tool for MPWTS were developed by utilizing relevant literature, including field studies, and implementing and comparing seven different MPWTS. The proposed System Life Cycle Evaluation (SLiCE) method uses these CTQs to evaluate the diversity in system performance and harmonize relevant performance with stakeholder preference via a selection tool. Agencies and field workers can use SLiCE results to inform and drive decision-making. The evaluation and selection tool also serves as a catalyst for communicating system performance, common design flaws, and stakeholder needs to system manufacturers. The SLiCE framework can be adopted into other emerging system technologies to communicate system performance over the life cycle of use.


Thoracic ultrasound competence for ultrasound-guided pleural procedures.
Focused thoracic ultrasound has become essential in the guidance and direction of pleural interventions to reduce unwanted complications and as a result now forms a crucial component of physician training. Current training standards along with assessment methods vary widely, and are often not robust enough to ensure adequate competence.This review assesses the current state of training and assessment of thoracic ultrasound competence in various settings, allowing comparison with alternative competency based programmes. Future directions for training and assessment of thoracic ultrasound competence are discussed.


Zika among international travellers presenting to GeoSentinel sites, 2012-2019: implications for clinical practice.
International travellers contribute to the rapid spread of Zika virus (ZIKV) and its sentinel identification globally. We describe ZIKV infections among international travellers seen at GeoSentinel sites with a focus on ZIKV acquired in the Americas and the Caribbean, describe countries of exposure and traveller characteristics, and assess ZIKV diagnostic testing by site. Records with an international travel-related diagnosis of confirmed or probable ZIKV from January 2012 through December 2019 reported to GeoSentinel with a recorded illness onset date were included to show reported cases over time. Records from March 2016 through December 2019 with an exposure region of the Americas or the Caribbean were included in the descriptive analysis. A survey was conducted to assess the availability, accessibility and utilization of ZIKV diagnostic tests at GeoSentinel sites. GeoSentinel sites reported 525 ZIKV cases from 2012 through 2019. Between 2012 and 2014, eight cases were reported, and all were acquired in Asia or Oceania. After 2014, most cases were acquired in the Americas or the Caribbean, a large decline in ZIKV cases occurred in 2018-19.Between March 2016 and December 2019, 423 patients acquired ZIKV in the Americas or the Caribbean, peak reporting to these regions occurred in 2016 [330 cases (78%)]. The median age was 36 years (range: 3-92); 63% were female. The most frequent region of exposure was the Caribbean (60%). Thirteen travellers were pregnant during or after travel; one had a sexually acquired ZIKV infection. There was one case of fetal anomaly and two travellers with Guillain-Barré syndrome. GeoSentinel sites reported various challenges to diagnose ZIKV effectively. ZIKV should remain a consideration for travellers returning from areas with risk of ZIKV transmission. Travellers should discuss their travel plans with their healthcare providers to ensure ZIKV prevention measures are taken.


P2X(7) receptors in the enteric nervous system of guinea-pig small intestine.
The P2X(7) purinergic receptor subtype has been cloned and emphasized as a prototypic P2Z receptor involved in neurotransmission in the central nervous system and ATP-mediated lysis of macrophages in the immune system. Less is known about the neurobiology of P2X(7) receptors in the enteric nervous system (ENS). We studied the distribution of the receptor with indirect immunofluorescence and used selective agonists and antagonists to analyze pharmacologic aspects of its electrophysiologic behavior as determined with intracellular "sharp" microelectrodes and patch-clamp recording methods in neurons identified morphologically by biocytin injection in the ENS. Application of ATP or 2'- (or-3'-) O-(4-benzoylbenzoyl) adenosine 5'-triphosphate (BzBzATP) activated an inward current in myenteric neurons. Brilliant blue G, a selective P2X(7) antagonist, suppressed the responses to both agonists. Potency of the antagonist was greatest (smaller IC(50)) for the current evoked by BzBzATP. The P2X(7) antagonists 1-[N,O-bis (1,5-isoquinolinesulfonyl)-N-methyl-l-tyrosyl]-4-piperazine (KN-62) and oxidized ATP also suppressed the BzBzATP-activated current. Micropressure application of BzBzATP evoked rapidly activating depolarizing responses in intracellular studies with "sharp" microelectrodes. Oxidized-ATP suppressed these responses in both myenteric and submucosal neurons. Rapidly activating depolarizing responses evoked by application of nicotinic, serotonergic 5-HT(3), or gamma-aminobutyric acid A (GABA(A)) receptor agonists were unaffected by brilliant blue G. Immunoreactivity for the P2X(7) receptor was widely distributed surrounding ganglion cell bodies and associated with nerve fibers in both myenteric and submucous plexuses. P2X(7) immunoreactivity was colocalized with synapsin and synaptophysin and surrounded ganglion cells that contained either calbindin, calretinin, neuropeptide Y, substance P, or nitric oxide synthase. The mucosa, submucosal blood vessels, and the circular muscle coat also showed P2X(7) receptor immunoreactivity.


Successful lung donation at the age of 6 weeks: Challenges and lessons learned.
A clinical case of successful procurement and transplantation of bilateral lungs from 6-week-old infant with sepsis secondary to bacterial meningitis is reported. Forty-one-day-old male infant (height 60 cm, weight 4 kg) died of cerebral edema secondary to Escherichia coli meningitis and bacteremia. Preretrieval assessment included the following: arterial gases; pO2 50.4 kPa (378 mm Hg), pCO2 4.9 kPa (37 mm Hg), on FiO2 100%, PEEP 5 cm H2 O. Fiberoptic bronchoscopy showed no secretions nor mucosal inflammation; CXR revealed clear lung fields and pleural spaces. Inspection revealed dense adhesions in pericardial cavity and purulent left hemithorax effusion (urgent Gram-stain came back as negative) but there was no consolidation in the lung. Good compliance of the lungs on inflation/deflation test was observed. The lungs were retrieved using the technique described. The recipient was a 4-month-old infant with alveolar capillary dysplasia and malaligned pulmonary veins. Implantation of the lungs was performed via bilateral thoracosternotomy on cardiopulmonary bypass, cooling to 30°C. Elective support with nitric oxide was used postoperatively. Two years after the transplantation, the recipient doing well with normal lung function. Lung procurement from a 6-week donor with infectious complications and prolonged ventilation is a challenging undertaking but can be successful and should be attempted whenever possible given the paucity of organs available for pediatric recipients.


Loss of integrity of thyroid morphology and function in children born to mothers with inadequately treated Graves' disease.
Central congenital hypothyroidism (CH-C) in neonates born to mothers with inadequately treated Graves' disease usually needs T(4) supplementation. The thyroid and its regulatory system have not yet been extensively studied after T(4) withdrawal, until we observed disintegrated thyroid glands in some patients. The aim was to study the occurrence and pathogenesis of disintegrated thyroid glands in CH-C patients. DESIGN, SETTING, PATIENTS, PARTICIPANTS: Thyroid function was measured and thyroid ultrasound imaging was performed in 13 children with CH-C due to inadequately treated maternal Graves' disease after T(4)-supplementation withdrawal (group Aa). In addition, thyroid ultrasound imaging was performed in six children with CH-C born to inadequately treated mothers with Graves' disease, in whom T(4) supplementation was not withdrawn yet (group Ab) or never initiated (group Ac), in six euthyroid children born to adequately treated mothers with Graves' disease (group B), and in 10 T(4)-supplemented children with CH-C as part of multiple pituitary hormone deficiency (group C). Thyroid function and aspect (volume, echogenicity, echotexture) were measured. In group A, five children had developed thyroidal hypothyroidism characterized by persistently elevated TSH concentrations and exaggerated TSH responses after TRH stimulation. In the majority of patients in groups A and C, thyroid echogenicity and volume were decreased, and echotexture was inhomogeneous. Thyroid ultrasound imaging was normal in group B children. Inadequately treated maternal Graves' disease not only may lead to CH-C but also carries an, until now, unrecognized risk of thyroid disintegration in the offspring as well. We speculate that insufficient TSH secretion due to excessive maternal-fetal thyroid hormone transfer inhibits physiological growth and development of the child's thyroid.


Non-contact video-based vital sign monitoring using ambient light and auto-regressive models.
Remote sensing of the reflectance photoplethysmogram using a video camera typically positioned 1 m away from the patient's face is a promising method for monitoring the vital signs of patients without attaching any electrodes or sensors to them. Most of the papers in the literature on non-contact vital sign monitoring report results on human volunteers in controlled environments. We have been able to obtain estimates of heart rate and respiratory rate and preliminary results on changes in oxygen saturation from double-monitored patients undergoing haemodialysis in the Oxford Kidney Unit. To achieve this, we have devised a novel method of cancelling out aliased frequency components caused by artificial light flicker, using auto-regressive (AR) modelling and pole cancellation. Secondly, we have been able to construct accurate maps of the spatial distribution of heart rate and respiratory rate information from the coefficients of the AR model. In stable sections with minimal patient motion, the mean absolute error between the camera-derived estimate of heart rate and the reference value from a pulse oximeter is similar to the mean absolute error between two pulse oximeter measurements at different sites (finger and earlobe). The activities of daily living affect the respiratory rate, but the camera-derived estimates of this parameter are at least as accurate as those derived from a thoracic expansion sensor (chest belt). During a period of obstructive sleep apnoea, we tracked changes in oxygen saturation using the ratio of normalized reflectance changes in two colour channels (red and blue), but this required calibration against the reference data from a pulse oximeter.


Seasonal losses of dissolved organic carbon and total dissolved solids from rice production systems in northern California.
Water quality concerns have arisen related to rice (Oryza sativa L.) field drain water, which has the potential to contribute large amounts of dissolved organic carbon (DOC) and total dissolved solids (TDS) to the Sacramento River. Field-scale losses of DOC or TDS have yet to be quantified. The objectives of this study were to evaluate the seasonal concentrations of DOC and TDS in rice field drain water and irrigation canals, quantify seasonal fluxes and flow-weighted (FW) concentrations of DOC and TDS, and determine the main drivers of DOC and TDS fluxes. Two rice fields with different straw management practices (incorporation vs. burning) were monitored at each of four locations in the Sacramento Valley. Fluxes of DOC ranged from 3.7 to 34.6 kg ha(-1) during the growing season (GS) and from 0 to 202 kg ha(-1) during the winter season (WS). Straw management had a significant interaction effect with season, as the greatest DOC concentrations were observed during winter flooding of straw incorporated fields. Fluxes and concentrations of TDS were not significantly affected by either straw management or season. Total seasonal water flux accounted for 90 and 88% of the variability in DOC flux during the GS and WS, respectively. Peak DOC concentrations occurred at the onset of drainflow; therefore, changes in irrigation management may reduce peak DOC concentrations and thereby DOC losses. However, the timing of peak DOC concentrations from rice fields suggest that rice field drainage water is not the cause of peak DOC concentrations in the Sacramento River.


Delivering sustainability therapy in sustainable development projects.
This paper explores the apparent contradiction between the 'linearity' of most Sustainable Development projects, with time-bound and defined outputs achieved at a fixed cost, and an implied 'circularity' of the theory whereby there is no 'end'. Projects usually have clear parameters within which they are implemented, and the inclusion of elements such as the need for accountability, measurable impact and 'value for money' have grown in importance. It could be argued that we live in a 'projectified' and therefore linear world. The paper explores the potential contradiction between 'linearity' and 'circularity', and suggests that one way around this is to frame the project within a form of the Kolb Learning Cycle heuristic. This will facilitate a rationalisation from those implementing the sustainable development project as to why decisions are being made and for whom. If these questions are opened up to the project stakeholders, including beneficiaries, then the Kolb cycle could encourage learning and understanding by all involved. It could also provide Sustainability Therapy to those trapped in processes, which they find orthogonal to their own perceptions. It is suggested that such learning, therapy and reflective practice should be a valid output of the sustainable development project, although typically the focus is only upon the final outputs and how they feed into policy. Ironically funders would be well advised to take a broader perspective in order to achieve true 'value for money' within such projects, even if learning is not an easily measurable or tangible outcome. These points are explored within the context of the wider literature and experience with a sustainable development project undertaken in Malta.


Adult Spinal Deformity: National Trends in the Presentation, Treatment, and Perioperative Outcomes From 2003 to 2010.
Retrospective review of a prospective database. To investigate adult spinal deformity (ASD) surgery outcome trends on a nationwide scale using the Nationwide Inpatient Sample (NIS) from 2003 to 2010. ASD patients ≥25 years from 2003 to 2010 in the NIS undergoing anterior, posterior, or combined surgical approaches were included. Fractures, 9+ levels fused, or any cancer were excluded. Patient demographics, hospital data, and procedure-related complications were evaluated. Yearly trends were analyzed using univariate analysis and linear regression modeling. Of 10,966 discharges, 1,952 were anterior, 6,524 were posterior, and 1,106 were combined. The total surgical ASD volume increased by 112.5% (p = .029), and both the average patient age (p < .001) and number of patients >65 years old significantly increased from 2003 to 2010 (p = .009). Anterior approach case volume decreased by 13.7% (p = .019), whereas that of combined increased by 22.7% (p = .047). Posterior case volume increased by 38.9% from 2003 to 2010, though insignificantly (p = .084). Total hospital charges for all approaches increased over the interval (p < .001). Total length of stay for all approaches decreased over the time interval (p < .005). Although the overall morbidity for all approaches increased by 22.7% (p < .001), mortality did not change (p = .817). The most common morbidities in 2003 were hemorrhagic anemia, accidental cut, puncture, perforation, or laceration during a procedure, and device-related complications, which persisted in 2010 with the exception of increased acute respiratory distress syndrome and pulmonary-related complications. For ASD surgery from 2003 to 2010, the volume of anterior approaches decreased, whereas posterior procedures did not change, and combined approaches increased. Total hospital charges increased for all considered procedures, length of hospital stay decreased, whereas operative patients were increasingly elderly, and more procedures were observed for patients >65 years old. For all approaches, morbidity increased whereas mortality did not change. Future study is required to develop methods to reduce morbidity and costs, thereby optimizing patient outcomes.


Implementation of the Healthy Workplace Participatory Program in a Retail Setting: A Feasibility Study and Framework for Evaluation.
Participatory methods used in Total Worker Health® programs have not been well studied, and little is known about what is needed to successfully implement these programs. We conducted a participatory health promotion program with grocery store workers using the Healthy Workplace Participatory Program (HWPP) from the Center for the Promotion of Health in the New England Workplace. We recruited a design team made up of six line-level workers and a steering committee with management and union representatives; a research team member facilitated the program. Using a formal evaluation framework, we measured program implementation including workplace context, fidelity to HWPP materials, design team and steering committee engagement, program outputs, and perceptions of the program. The HWPP was moderately successful in this setting, but required a substantial amount of worker and facilitator time. Design team members did not have the skills needed to move through the process and the steering committee did not offer adequate support to compensate for the team's shortfall. The evaluation framework provided a simple and practical method for identifying barriers to program delivery. Future studies should address these barriers to delivery and explore translation of this program to other settings.


The development and validation of the Evidence-Based Practice Questionnaire: Japanese version.
To develop and test the validity and reliability of a Japanese version of the Evidence-Based Practice Questionnaire. The Evidence-Based Practice Questionnaire was translated into Japanese, and a back-translation was performed. In March 2016, the survey was conducted among 843 nurses in 2 university hospitals in Japan. After 2 weeks, the same questionnaire for test-retest reliability was distributed to 197 nurses. We evaluated construct validity by factor analysis: convergent, discriminant, criterion-based and known-group validity, and reliability (internal consistency via Cronbach' alpha and test-retest reliability). A total of 533 (66%) of nurses who agreed to our study returned the completed questionnaire, and 108 nurses (55%) agreed to the test-retest. Factor analysis confirmed that the original model of the Evidence-Based Practices Questionnaire was not statistically appropriate for Japanese nurses. Therefore, the final version of the Japanese version adopted 18 items and 4 subscales including practice, attitudes, and knowledge/skills related to research and practice. In the final version, the validity and reliability were moderate (Cronbach alpha = .90). The Japanese version of the Evidence-Based Practices Questionnaire is a reliable and valid tool and can be used to assess evidence-based practice, attitudes, and research knowledge/skills in Japan.


Differential regulation of three genes encoding glutathione S-transferases in Schizosaccharomyces pombe.
Glutathione S-transferases (GSTs) are detoxifying enzymes that catalyze the conjugation of glutathione with a variety of reactive electrophilic compounds. Three GST genes were previously characterized in the fission yeast Schizosaccharomyces pombe. In this work, we examined the transcriptional regulation of these genes using individual GST-lacZ fusions and RT-PCR. Basal synthesis of beta-galactosidase from the GSTII-lacZ fusion was higher than from the GSTI-lacZ and GSTIII-lacZ fusion. Diethylmaleate (0.2 mM) greatly enhanced the synthesis of beta-galactosidase from the GSTII-lacZ fusion, but did not affect synthesis from the other two fusion genes. A switch to 0.3% glucose or 0.3% sucrose as sole carbon source enhanced expression from the GSTIII-lacZ fusion gene, while sodium nitroprusside (1.5 mM), tert-butylhydroquinone (0.2 mM), and L-buthionine-[S,R]-sulfoximine (0.01 mM) increased expression of the GSTII gene. The effects of these agents on GST mRNA levels were confirmed by measurements employing RT-PCR. Our results suggest that transcription of the three S. pombe GST genes is subjected to differential regulation under various stress conditions, and may be linked to their different physiological functions.


Diffusivity of gases in liquids.
Diffusion coefficients of H(2), Ne, N(2), Ar, CH(4), Cl(2), CF(4), C(2)H(6), SF(6), I(2), and isotopic CCl(4), all in CCl(4), determined at atmospheric pressure, are linear functions of temperature, converging to zero at the temperature where CCl(4) ceases to be fluid. The slopes of these lines increase with decreasing molecular cross-section of the diffusants, and with increasing entropy of expansion of the diffusants in CCl(4). Diffusivities in (C(4)F(9))(2)N, whose molecules are very large and three-armed, do not converge as temperature is decreased. Molecules of H(2), Ne, and, to a lesser extent, Ar, are able to diffuse in (C(4)F(9))(2)N even at temperatures where fluidity is low.


A longitudinal field trial of the effect of biotin on lameness in dairy cows.
A longitudinal prospective intervention study investigated the effect of biotin supplementation on the incidence (new cases per day) of visible lameness in milking cows and heifers on five commercial farms in Gloucestershire, United Kingdom. The trial lasted from June 1997 to April 1999. Each farm participated in the trial for 18 mo. Within each herd the cows were randomly allocated to either receive a supplement of 20 mg of biotin per day or not. All cows were run as one herd on each farm. When a lame cow was identified, the farmer called one of six veterinarians to examine and treat the affected animal; findings were recorded on a standard form. A veterinarian also carried out a bimonthly locomotion assessment to ensure that all lame cows were diagnosed. There were a total of 900 cows, 1120 cow years, in the trial. The overall incidence rate of lameness (per 100 cows per year) was 68.9, with a range of 31.6 to 111.5 per farm. The incidence rates of the four most frequently reported causes of lameness were sole ulcer, 13.8; white line separation, 12.7; digital dermatitis, 12.0; and interdigital necrobacillosis, 7.1 per 100 cows per year. There was a significant difference in the incidence rate of these four lesions between supplemented and unsupplemented cows on two of the five farms, with a significant decrease in lameness in the cows supplemented with biotin. When all the farms were pooled, the risk of lameness caused by white line separation in cattle supplemented with biotin was approximately halved (Cox proportional hazard survival analysis hazard ratio = 0.57).


Appendage expression driven by the Hoxd Global Control Region is an ancient gnathostome feature.
The evolutionary transition of the fins of fish into tetrapod limbs involved genetic changes to developmental systems that resulted in novel skeletal patterns and functions. Approaches to understanding this issue have entailed the search for antecedents of limb structure in fossils, genes, and embryos. Comparative genetic analyses have produced ambiguous results: although studies of posterior Hox genes from homology group 13 (Hoxa-13 and Hoxd-13) reveal similarities in gene expression between the distal segments of fins and limbs, this functional homology has not been supported by genomic comparisons of the activity of their cis-regulatory elements, namely the Hoxd Global Control Region. Here, we show that cis-regulatory elements driving Hoxd gene expression in distal limbs are present in fish. Using an interspecies transgenesis approach, we find functional conservation between gnathostome Hoxd enhancers, demonstrating that orthologous sequences from tetrapods, zebrafish and skate can drive reporter gene expression in mouse limbs and zebrafish fins. Our results support the notion that some of the novelties associated with tetrapod limbs arose by modification of deeply conserved cis- and trans-acting mechanisms of Hox regulation in gnathostomes.


Pamidronate Treatment of Osteonecrosis of the Hip in Young Male.
Aseptic osteonecrosis of the hip is a clinical entity in which the necrotic process of the bone leads to pain and progressive disability. Pamidronate seems to reduce drastically the activation of the osteoclasts so that it can be useful only in the early stage of the disease, delaying the time of bone collapsing. A 27-years-old male was treated with pamidronate for three consecutive days every four weeks. After three months the patient came back at control showing a marked improvement in clinical condition, referred full recover from pain and dysmotility with improvement of the quality of life, which was confirmed by the result of MRI he had for control. We reported a case of aseptic osteonecrosis of the hip which was successfully treated pamidronate at dosage of 45 mg.


Genomic Changes Associated with the Evolutionary Transitions of Nostoc to a Plant Symbiont.
Cyanobacteria belonging to the genus Nostoc comprise free-living strains and also facultative plant symbionts. Symbiotic strains can enter into symbiosis with taxonomically diverse range of host plants. Little is known about genomic changes associated with evolutionary transition of Nostoc from free-living to plant symbiont. Here, we compared the genomes derived from 11 symbiotic Nostoc strains isolated from different host plants and infer phylogenetic relationships between strains. Phylogenetic reconstructions of 89 Nostocales showed that symbiotic Nostoc strains with a broad host range, entering epiphytic and intracellular or extracellular endophytic interactions, form a monophyletic clade indicating a common evolutionary history. A polyphyletic origin was found for Nostoc strains which enter only extracellular symbioses, and inference of transfer events implied that this trait was likely acquired several times in the evolution of the Nostocales. Symbiotic Nostoc strains showed enriched functions in transport and metabolism of organic sulfur, chemotaxis and motility, as well as the uptake of phosphate, branched-chain amino acids, and ammonium. The genomes of the intracellular clade differ from that of other Nostoc strains, with a gain/enrichment of genes encoding proteins to generate l-methionine from sulfite and pathways for the degradation of the plant metabolites vanillin and vanillate, and of the macromolecule xylan present in plant cell walls. These compounds could function as C-sources for members of the intracellular clade. Molecular clock analysis indicated that the intracellular clade emerged ca. 600 Ma, suggesting that intracellular Nostoc symbioses predate the origin of land plants and the emergence of their extant hosts.


Guide RNAs: A Glimpse at the Sequences that Drive CRISPR-Cas Systems.
CRISPR-Cas systems provide adaptive immunity in bacteria and archaea. Although there are two main classes of CRISPR-Cas systems defined by gene content, interfering RNA biogenesis, and effector proteins, Type II systems have recently been exploited on a broad scale to develop next-generation genetic engineering and genome-editing tools. Conveniently, Type II systems are streamlined and rely on a single protein, Cas9, and a guide RNA molecule, comprised of a CRISPR RNA (crRNA) and trans-acting CRISPR RNA (tracrRNA), to achieve effective and programmable nucleic acid targeting and cleavage. Currently, most commercially available Cas9-based genome-editing tools use the CRISPR-Cas system from Streptococcus pyogenes (SpyCas9), although many orthogonal Type II systems are available for diverse and multiplexable genome engineering applications. Here, we discuss the biological significance of Type II CRISPR-Cas elements, including the tracrRNA, crRNA, Cas9, and protospacer-adjacent motif (PAM), and look at the native function of these elements to understand how they can be engineered, enhanced, and optimized for genome editing applications. Additionally, we discuss the basis for orthogonal Cas9 and guide RNA systems that would allow researchers to concurrently use multiple Cas9-based systems for different purposes. Understanding the native function of endogenous Type II CRISPR-Cas systems can lead to new Cas9 tool development to expand the genetic manipulation toolbox.


Clinical, genetic and DNA repair studies on a consecutive series of patients with xeroderma pigmentosum.
We report clinical, genetic and biochemical findings in 13 families with the photosensitive genodermatosis, xeroderma pigmentosum. All patients had a defect in repair of DNA damage provoked by ultraviolet radiation. Eleven patients and their three affected sibs were defective in the excision repair of UVR induced DNA lesions while the other two were defective in post-replication repair. One in the former group was diagnosed prior to the development of permanent skin abnormalities and preventive measures succeeded for almost five years in maintaining a normal appearing skin. In addition, two cases were diagnosed prenatally and aborted therapeutically. Some patients' parents showed slightly reduced repair of UVR induced DNA damage. In xeroderma pigmentosum (XP), the defect in the excision of DNA lesions appears to be due to homozygosity for one of at least seven different mutations and, accordingly, XP patients can be assigned to seven so-called complementation groups, A to G. Of these, groups A, C and D are the most common. Somatic cell fusion allowed three of the families reported here to be assigned to group A, four to group C and four to group D. Fibroblasts of patients from these three groups were shown to differ not only in the degree and kinetics of their residual DNA repair but also in the kinetics with which their defect is complemented by fusion with normal or XP cells of other groups. This confirms that mutations of different genes play a role in XP and provides a basis for understanding how such genes interact to secure repair of DNA lesions in normal cells. We discuss the phenotype of XP from different complementation groups in relation to the severe neurological abnormalities which may develop and must be considered in genetic counselling. We also discuss the biochemical anomalies of XP and the cellular effects of physical and chemical agents which damage DNA. In the practical management of XP, the importance of early differential diagnosis and prompt initiation of treatment is emphasized. Lastly we review the relationship between DNA repair and skin cancer in XP.


Recent advances in cochlear blood flow measurements.
Changes in blood flow to the inner ear have been thought to influence or underlie a number of cochlear diseases, including some forms of noise-induced hearing loss, sudden hearing loss, and Meniere's disease. Recently, important advances have been made in two technologies for the study of cochlear blood flow. The first is in the area of vital microscopic studies of cochlear microcirculation, and the second is based on the introduction of laser technology in the form of laser Doppler flowmetry. In this report, measurements are given of changes in cochlear circulation caused by carbon dioxide breathing, intravenous phenylephrine injection, systemic hemodilution, positive end expiratory pressure, and direct electrical stimulation of the cochlea. From these changes, we observe that cochlear blood circulation responds to systemic blood pressure alterations and is subject to local flow control mechanisms. Linearity and speed of response of the laser Doppler instrumentation also are shown. These advances show promise for contributing to our knowledge of control mechanisms of inner ear blood flow and for revealing the influence of various pharmacologic agents of potential clinical value.


Linked regularities in the development and evolution of mammalian brains.
Analysis of data collected on 131 species of primates, bats, and insectivores showed that the sizes of brain components, from medulla to forebrain, are highly predictable from absolute brain size by a nonlinear function. The order of neurogenesis was found to be highly conserved across a wide range of mammals and to correlate with the relative enlargement of structures as brain size increases, with disproportionately large growth occurring in late-generated structures. Because the order of neurogenesis is conserved, the most likely brain alteration resulting from selection for any behavioral ability may be a coordinated enlargement of the entire nonolfactory brain.


Cost-effectiveness of endovascular therapy in the surgical management of cerebral arteriovenous malformations.
To determine the economic effect of endovascular therapy in conjunction with surgery for cerebral arteriovenous malformations. Twenty-five patients with arteriovenous malformations treated with embolization and surgical excision or embolization alone were compared with reported results in 475 patients who underwent surgery only. Respective mean morbidity and mortality rates were calculated and a cost-effectiveness analysis was performed in terms of costs of hospitalization, professional fees, and other direct procedural and indirect costs. Quality-adjusted life-years saved were also calculated. The net effective treatment cost per cure was $71 366 (in 1992 dollars) for embolization and surgery compared with $78 506 for surgery alone. This resulted in a 9% average savings per treated patient. Cost per quality-adjusted life-year calculations resulted in a cost of $6734 for embolization and surgery and $9814 for surgical treatment alone, with savings as high as 34% when endovascular therapy was used. Endovascular therapy in conjunction with surgery resulted in significant economic benefits for treatment of cerebral arteriovenous malformations.


The effect of surgical synovectomy and radiotherapy on the rate of recurrence of pigmented villonodular synovitis of the knee: an individual patient meta-analysis.
Pigmented villonodular synovitis (PVNS) is a rare proliferative process of the synovium which most commonly affects the knee and occurs in either a localised (LPVNS) or a diffuse form (DPVNS). The effect of different methods of surgical synovectomy and adjuvant radiotherapy on the rate of recurrence is unclear. We conducted a systematic review and identified 35 observational studies in English which reported the use of surgical synovectomy to treat PVNS of the knee. A meta-analysis included 630 patients, 137 (21.8%) of whom had a recurrence after synovectomy. For patients with DPVNS, low-quality evidence found that the rate of recurrence was reduced by both open synovectomy (odds ration (OR) = 0.47; 95% CI 0.25 to 0.90; p = 0.024) and combined open and arthroscopic synovectomy (OR = 0.19, 95% CI = 0.06 to 0.58; p = 0.003) compared with arthroscopic surgery. Very low-quality evidence found that the rate of recurrence of DPVNS was reduced by peri-operative radiotherapy (OR = 0.31, 95% CI 0.14 to 0.70; p = 0.01). Very low-quality evidence suggested that the rate of recurrence of LPVNS was not related to the surgical approach. This meta-analysis suggests that open synovectomy or synovectomy combined with peri-operative radiotherapy for DPVNS is associated with a reduced rate of recurrence. Large long-term prospective multicentre observational studies, with a focus on both rate of recurrence and function, are required to confirm these findings.


Benchmark data sets for structure-based computational target prediction.
Structure-based computational target prediction methods identify potential targets for a bioactive compound. Methods based on protein-ligand docking so far face many challenges, where the greatest probably is the ranking of true targets in a large data set of protein structures. Currently, no standard data sets for evaluation exist, rendering comparison and demonstration of improvements of methods cumbersome. Therefore, we propose two data sets and evaluation strategies for a meaningful evaluation of new target prediction methods, i.e., a small data set consisting of three target classes for detailed proof-of-concept and selectivity studies and a large data set consisting of 7992 protein structures and 72 drug-like ligands allowing statistical evaluation with performance metrics on a drug-like chemical space. Both data sets are built from openly available resources, and any information needed to perform the described experiments is reported. We describe the composition of the data sets, the setup of screening experiments, and the evaluation strategy. Performance metrics capable to measure the early recognition of enrichments like AUC, BEDROC, and NSLR are proposed. We apply a sequence-based target prediction method to the large data set to analyze its content of nontrivial evaluation cases. The proposed data sets are used for method evaluation of our new inverse screening method iRAISE. The small data set reveals the method's capability and limitations to selectively distinguish between rather similar protein structures. The large data set simulates real target identification scenarios. iRAISE achieves in 55% excellent or good enrichment a median AUC of 0.67 and RMSDs below 2.0 Å for 74% and was able to predict the first true target in 59 out of 72 cases in the top 2% of the protein data set of about 8000 structures.


Development of continuous surfactin production from potato process effluent by Bacillus subtilis in an airlift reactor.
The biosurfactant surfactin has the potential to aid in the recovery of subsurface organic contaminants (environmental remediation) or crude oils (oil recovery). However, high medium and purification costs limit its use in these high-volume applications. In previous work, we showed that surfactin can be produced from an inexpensive low-solids (LS) potato process effluent with minimal amendments or pretreatments. Previous research has also shown that 95% or more of the surfactin in Bacillus subtilis cultures can be recovered by foam fractionation. In this work, we present the results of research to integrate surfactin production with foam fractionation. Experiments were performed in an airlift reactor, with continuous collection of the foam through a tube at the top of the column. Preliminary results using both purified potato starch and unamended low-solids potato process effluent as substrates for surfactin production indicate that the process is oxygen limited and that recalcitrant indigenous bacteria in the potato process effluent may hamper continuous surfactin production.


Aseptic meningitis in the newborn and young infant.
When a toxic newborn or young infant presents with fever and lethargy or irritability, it is important to consider the diagnosis of meningitis even if the classic localizing signs and symptoms are absent. Cerebrospinal fluid should be obtained (unless lumbar puncture is clinically contraindicated) to enable initial therapy to be planned. Initial results of cerebrospinal fluid testing may not conclusively differentiate between aseptic and bacterial meningitis, and antimicrobial therapy for all likely organisms should be instituted until definitive culture results are available. Comprehensive therapy, including antibacterial and antiviral agents, should continue until a cause is identified and more specific therapy is initiated, an etiology is excluded or the patient improves considerably and the course of antimicrobial therapy is completed. Group B streptococcus is the most common bacterial etiologic agent in cases of meningitis that occur during the first month after birth. Etiologies of aseptic meningitis include viral infection, partially treated bacterial meningitis, congenital infections, drug reactions, postvaccination complications, systemic diseases and malignancy. Long-term sequelae of meningitis include neuromuscular impairments, learning disabilities and hearing loss. Prompt diagnosis and treatment are essential to improved outcome.


Anthocyanin-flavanol condensation products from black currant (Ribes nigrum L.).
Putative flavanol-anthocyanin condensation products were detected in a polyphenol-rich concentrate from black currant (Ribes nigrum L.). These compounds had UV-vis spectra similar to those of delphinidin-3-O-rutinoside and cyanidin-3-O-rutinoside, but eluted before all previously described anthocyanins on reversed phase HPLC. Mass spectrometric data indicated that they were rutinoside derivatives of novel aglycons 304 amu greater than delphinidin and cyanidin, respectively. The compounds were partly purified by semipreparative HPLC and gave MS and MS2 spectra consistent with anthocyanin rutinosides covalently linked to epigallocatechin or gallocatechin. These compounds are similar in structure to compounds thought to influence color and quality in red wines and strawberry juice products. There was also evidence for the presence of a range of other flavanol-anthocyanin condensation products. The compounds were present at differing levels in juices of 10 black currant varieties, which were roughly correlated to the content of the parent anthocyanins. The flavanol-anthocyanin products were present in polyphenol-enriched concentrates obtained by solid phase extraction, in commercially produced concentrates, and in fresh extracts of black currants. This suggests that the compounds were not artifacts formed during concentration or purification. However, differences in their comparative contents may be related to the lability of the parent anthocyanins during processing. Although present at low levels, the flavanol-anthocyanin products may influence color or quality parameters of black currant juices, and they may confer enhanced stability to the biological activities reported for their anthocyanin parents.


Murine pancreatic adenocarcinoma reduces Ikaros expression and disrupts T cell homeostasis.
Maintenance of T cell immune homeostasis is critical for adequate anti-tumor immunity. The transcription factor Ikaros is essential for lymphocyte development including T cells. Alterations in Ikaros expression occur in blood malignancies in humans and mice. In this study, we investigated the role of Ikaros in regulating T cell immune balance in pancreatic cancer mouse models. Using our Panc02 tumor-bearing (TB) mouse model, western blot analysis revealed a reduction in Ikaros proteins while qRT-PCR showed no differences in Ikaros mRNA levels in TB splenocytes compared to control. Treatment of naïve splenocytes with the proteasomal inhibitor, MG132, stabilized Ikaros expression and prevented Ikaros downregulation by Panc02 cells, in vitro. Western blot analyses showed a reduction in protein phosphatase 1 (PP1) and protein kinase CK2 expression in TB splenocytes while CK2 activity was increased. Immunofluorescence microscopy revealed altered punctate staining of Ikaros in TB splenocytes. Flow cytometry revealed a significant decrease in effector CD4+ and CD8+ T cell percentages but increased CD4+CD25+ regulatory T cells in TB splenocytes. Similar alterations in T cell percentages, as well as reduced Ikaros and CK2 but not PP1 expression, were observed in a transgenic, triple mutant (TrM) pancreatic cancer model. Ikaros expression was also reduced in enriched TB CD3+ T cells. MG132 treatment of naïve CD3+ T cells stabilized Ikaros expression in the presence of Panc02 cells. Western blots showed reduced PP1 and CK2 expression in TB CD3+ T cells. The results of this study suggest that the pancreatic tumor microenvironment may cause proteasomal degradation of Ikaros, possibly via dysregulation of PP1 and CK2 expression and activity, respectively. This loss of Ikaros expression may contribute to an imbalance in T cell percentages. Ikaros may potentially be a therapeutic target to restore T cell homeostasis in pancreatic cancer hosts, which may be critical for effective anti-tumor immunity.


A framework for list representation, enabling list stabilization through incorporation of gene exchangeabilities.
Analysis of multivariate data sets from, for example, microarray studies frequently results in lists of genes which are associated with some response of interest. The biological interpretation is often complicated by the statistical instability of the obtained gene lists, which may partly be due to the functional redundancy among genes, implying that multiple genes can play exchangeable roles in the cell. In this paper, we use the concept of exchangeability of random variables to model this functional redundancy and thereby account for the instability. We present a flexible framework to incorporate the exchangeability into the representation of lists. The proposed framework supports straightforward comparison between any 2 lists. It can also be used to generate new more stable gene rankings incorporating more information from the experimental data. Using 2 microarray data sets, we show that the proposed method provides more robust gene rankings than existing methods with respect to sampling variations, without compromising the biological significance of the rankings.


Huntingtin controls neurotrophic support and survival of neurons by enhancing BDNF vesicular transport along microtubules.
Polyglutamine expansion (polyQ) in the protein huntingtin is pathogenic and responsible for the neuronal toxicity associated with Huntington's disease (HD). Although wild-type huntingtin possesses antiapoptotic properties, the relationship between the neuroprotective functions of huntingtin and pathogenesis of HD remains unclear. Here, we show that huntingtin specifically enhances vesicular transport of brain-derived neurotrophic factor (BDNF) along microtubules. Huntingtin-mediated transport involves huntingtin-associated protein-1 (HAP1) and the p150(Glued) subunit of dynactin, an essential component of molecular motors. BDNF transport is attenuated both in the disease context and by reducing the levels of wild-type huntingtin. The alteration of the huntingtin/HAP1/p150(Glued) complex correlates with reduced association of motor proteins with microtubules. Finally, we find that the polyQ-huntingtin-induced transport deficit results in the loss of neurotrophic support and neuronal toxicity. Our findings indicate that a key role of huntingtin is to promote BDNF transport and suggest that loss of this function might contribute to pathogenesis.


Contesting heteronormativity: the fight for lesbian, gay, bisexual and transgender recognition in India and Vietnam.
Recent public debates about sexuality in India and Vietnam have brought the rights of lesbian, gay, bisexual and transgender people sharply into focus. Drawing on legal documents, secondary sources and ethnographic fieldwork conducted in the urban centres of Delhi and Hanoi, this article shows how the efforts of civil society organisations dedicated to the fight for lesbian, gay, bisexual and transgender rights have had different consequences in these two Asian contexts. The paper considers how these organisations navigated government regulations about their formation and activities, as well as the funding priorities of national and international agencies. The HIV epidemic has had devastating consequences for gay men and other men who have sex with men, and has been highly stigmatising. As a sad irony, the epidemic has provided at the same time a strategic entry point for organisations to struggle for lesbian, gay, bisexual and transgender recognition. This paper examines how the fight for lesbian, gay, bisexual and transgender recognition has been doubly framed through health-based and rights-based approaches and how the struggle for recognition has positioned lesbian, gay, bisexual and transgender people in India and Vietnam differently.


Influence of the organic compounds on the ecotoxicity in the treatment of foundry sludge and olive mill waste.
The study of the ecotoxicity in two industrial waste materials and the relationships with the organic parameters has been conducted. Foundry sludge and olive mill waste have been used as industrial waste materials with organic or mixed character. Stabilisation/solidification (S/S) and advanced oxidation (AOP) processes have been carried out in order to treat both foundry sludge and olive mill waste. Analysis of ecotoxicity, total organic carbon, COD and phenol index have been evaluated in the untreated waste and end-products. The results of the treated samples allow obtaining the best formulations in both processes. The best formulations in the immobilisation process have been obtained with Portland cement and black carbon, activated carbon or foundry sand ashes. In the AOP process, ozone concentrations above 35 mg/l and reaction times equal to 120 minutes have been the optimal variables. The relationships between the organic parameters and the ecotoxicity of the samples have been studied in this paper. Furthermore, the global organic parameters have been studied in relation to the phenolic compounds. Lineal and logarithmic expressions have been obtained between the total organic carbon and phenol index and the ecotoxicity of the samples related to the organic parameters, respectively. Ecotoxicity of the samples with Vibrio fischeri is recommended as a very promising biotest for the study of the characterisation and the evaluation of the treatment of organic and mixed character waste and total organic carbon is recommended as global organic parameter in the treatment of foundry sludge.


Rock-scissors-paper and the survival of the weakest.
In the children's game of rock-scissors-paper, players each choose one of three strategies. A rock beats a pair of scissors, scissors beat a sheet of paper and paper beats a rock, so the strategies form a competitive cycle. Although cycles in competitive ability appear to be reasonably rare among terrestrial plants, they are common among marine sessile organisms and have been reported in other contexts. Here we consider a system with three species in a competitive loop and show that this simple ecology exhibits two counter-intuitive phenomena. First, the species that is least competitive is expected to have the largest population and, where there are oscillations in a finite population, to be the least likely to die out. As a consequence an apparent weakening of a species leads to an increase in its population. Second, evolution favours the most competitive individuals within a species, which leads to a decline in its population. This is analogous to the tragedy of the commons, but here, rather than leading to a collapse, the 'tragedy' acts to maintain diversity.


Toward Psychoinformatics: Computer Science Meets Psychology.
The present paper provides insight into an emerging research discipline called Psychoinformatics. In the context of Psychoinformatics, we emphasize the cooperation between the disciplines of psychology and computer science in handling large data sets derived from heavily used devices, such as smartphones or online social network sites, in order to shed light on a large number of psychological traits, including personality and mood. New challenges await psychologists in light of the resulting "Big Data" sets, because classic psychological methods will only in part be able to analyze this data derived from ubiquitous mobile devices, as well as other everyday technologies. As a consequence, psychologists must enrich their scientific methods through the inclusion of methods from informatics. The paper provides a brief review of one area of this research field, dealing mainly with social networks and smartphones. Moreover, we highlight how data derived from Psychoinformatics can be combined in a meaningful way with data from human neuroscience. We close the paper with some observations of areas for future research and problems that require consideration within this new discipline.


Maternal bile acid transporter deficiency promotes neonatal demise.
Intrahepatic cholestasis of pregnancy (ICP) is associated with adverse neonatal survival and is estimated to impact between 0.4 and 5% of pregnancies worldwide. Here we show that maternal cholestasis (due to Abcb11 deficiency) produces neonatal death among all offspring within 24 h of birth due to atelectasis-producing pulmonary hypoxia, which recapitulates the neonatal respiratory distress of human ICP. Neonates of Abcb11-deficient mothers have elevated pulmonary bile acids and altered pulmonary surfactant structure. Maternal absence of Nr1i2 superimposed on Abcb11 deficiency strongly reduces maternal serum bile acid concentrations and increases neonatal survival. We identify pulmonary bile acids as a key factor in the disruption of the structure of pulmonary surfactant in neonates of ICP. These findings have important implications for neonatal respiratory failure, especially when maternal bile acids are elevated during pregnancy, and highlight potential pathways and targets amenable to therapeutic intervention to ameliorate this condition.


Potential nitrification and denitrification on different surfaces in a constructed treatment wetland.
Improved understanding of the importance of different surfaces in supporting attached nitrifying and denitrifying bacteria is essential if we are to optimize the N removal capacity of treatment wetlands. The aim of this study was therefore to examine the nitrifying and denitrifying capacity of different surfaces in a constructed treatment wetland and to assess the relative importance of these surfaces for overall N removal in the wetland. Intact sediment cores, old pine and spruce twigs, shoots of Eurasian watermilfoil (Myriophyllum spicatum L.), and filamentous macro-algae were collected in July and November 1999 in two basins of the wetland system. One of the basins had been constructed on land that contained lots of wood debris, particularly twigs of coniferous trees. Potential nitrification was measured using the isotope-dilution technique, and potential denitrification was determined using the acetylene-inhibition technique in laboratory microcosm incubations. Nitrification rates were highest on the twigs. These rates were three and 100 times higher than in the sediment and on Eurasian watermilfoil, respectively. Potential denitrification rates were highest in the sediment. These rates were three times higher than on the twigs and 40 times higher than on Eurasian watermilfoil. The distribution of denitrifying bacteria was most likely due to the availability of organic material, with higher denitrification rates in the sediment than on surfaces in the water column. Our results indicate that denitrification, and particularly nitrification, in treatment wetlands could be significantly increased by addition of surfaces such as twigs.


Bifenthrin-induced neurotoxicity in rats: involvement of oxidative stress.
Extensive use of synthetic pyrethroids has resulted in serious human health issues. Induction of oxidative stress is an important mechanism of action of most pesticides including pyrethroids. In the present study, we have elucidated the possible role of oxidative stress in bifenthrin-induced neurotoxicity. Adult male Wistar rats were administered bifenthrin (3.5 and 7 mg per kg body weight p.o.) for 30 days. Behavioral studies were conducted on a set of randomly selected rats from each treatment group after completion of treatment. Neurochemical parameters were assessed 24 h after the last dose was administered. The selected behavioral and neurochemical endpoints were also assessed 15 days after cessation of exposure to reveal whether the neurobehavioral changes produced by bifenthrin were temporary or permanent. Deficits in motor activity, motor incoordination, and cognitive impairment were observed after exposure to bifenthrin. Levels of biogenic amines viz. dopamine (DA) and its metabolites, i.e. 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), epinephrine (EPN), norepinephrine (NE), and serotonin (5-HT) altered in the frontal cortex, corpus striatum, and hippocampus of bifenthrin-treated rats. A decrease in the activity of acetylcholinesterase (AChE) occurred in all regions of the brain. Both doses of bifenthrin significantly induced lipid peroxidation (LPO) and increased protein carbonyl levels in the frontal cortex, corpus striatum, and hippocampus of rats. The activities of antioxidant enzymes, i.e. catalase, superoxide dismutase, and glutathione peroxidase, were also suppressed in all selected regions of the brain. A trend of recovery was, however, observed in all the behavioral and neurochemical endpoints 15 days after withdrawal of exposure. Oxidative stress seems to play an important role in bifenthrin-induced neurotoxicity. Our study suggests that long-term exposure to these compounds can produce detrimental effects.


Prevalence of atrial fibrillation in the general population and in high-risk groups: the ECHOES study.
To establish the prevalence of atrial fibrillation (AF) in the general population in the UK, and in those with risk factors. The prevalence of AF on electrocardiography was established in prospectively selected groups: 3960 randomly selected from the population, aged 45+; 782 with a previous diagnosis of heart failure; and 1062 with a record of myocardial infarction, hypertension, angina, or diabetes. Patients were also assessed clinically and with echocardiography. Mortality was tracked for 8 years. Atrial fibrillation was found in 78 of the random population sample (2.0%). Prevalence was 1.6% in women and 2.4% in men, rising with age from 0.2% in those aged 45-54 to 8.0% in those aged 75 and older. Half of all cases were in patients aged 75 and older. Only 23 of the 78 (29.5%) of those in AF took warfarin. Of the 782 patients, 175 (22.4%) with a diagnosis of heart failure were in AF, with normal left ventricular function in 95 (54.3%) of these. Atrial fibrillation was found in 14 of the 244 (5.7%) of those with a history of myocardial infarction, 15 of the 388 (3.9%) of those with hypertension, 15 of the 321 (4.7%) of those with angina, and 11 of the 208 (5.3%) of diabetics. Adjusting for age and sex, mortality was 1.57 times higher for those in AF. Atrial fibrillation is common in the elderly and those with clinical risk factors. Screening these groups would identify many with AF. Use of anticoagulation was low at the time of the initial assessments in the late 1990s; practice may have changed recently.


Reproductive performance after the application of hyaluronic acid gel after dilation and curettage in women who have experienced at least one previous curettage: long-term results of a multicenter prospective randomized trial.
To examine whether intrauterine application of auto-cross-linked hyaluronic acid (ACP) gel after dilation and curettage (D&C) improves reproductive performance. Multicenter prospective randomized trial. One university and seven university-affiliated teaching hospitals. A total of 149 women who have experienced a miscarriage at <14 weeks of gestation, with at least one previous D&C for miscarriage or termination of pregnancy, were randomly assigned to either D&C plus ACP gel (intervention group) or D&C alone (control group). A hysteroscopy was performed 8-12 weeks after the procedure. Where intrauterine adhesions (IUAs) were found, adhesiolysis was executed. Participants received questionnaires 3, 6, and 12 months after the initial D&C procedure with questions regarding menstrual pattern, conception, and reproductive outcome. Reproductive outcomes during the 12 months after the D&C procedure. Outcomes were available for 149 women: 77 in the intervention group and 72 in the control group. The conception rates were, respectively, 71% (46/64) and 59.7% (40/67). There was no significant difference in the miscarriage, ongoing pregnancy, and live birth rates. The median times to conception were 5.5 and 7.1 months, respectively. Reproductive performance was similar in both groups although there was a significant difference in the prevalence of IUAs. Given the favorable reproductive outcome in this underpowered study and the fact that adhesiolysis was executed, no solid conclusion can be drawn. Future studies are needed to evaluate the effect of ACP gel on fertility and reproductive outcomes. NTR 3120.


Inter-cohort competition drives density dependence and selective mortality in a marine fish.
For organisms with complex life cycles, the transition between life stages and between habitats can act as a significant demographic and selective bottleneck. In particular, competition with older and larger conspecifics and heterospecifics may influence the number and characteristics of individuals successfully making the transition. We investigated whether the availability of enemy-free space mediated the interaction between adult goldspot gobies (Gnatholepis thompsoni), a common tropical reef fish, and juvenile conspecifics that had recently settled from the plankton. We added rocks, which provide refuge from predators, to one-half of each of five entire coral reefs in the Bahamas and measured the survival and growth of recent settlers in relation to adult goby densities. We also evaluated whether mortality was selective with respect to three larval traits (age at settlement, size at settlement, and presettlement growth rate) and measured the influence of refuge availability and adult goby density on selection intensity. Selective mortality was measured by comparing larval traits of newly settled gobies (< or = 5 d postsettlement) with those of survivors (2-3 week postsettlement juveniles). We detected a negative relationship between juvenile survival and adult goby density in both low- and high-refuge habitats, though experimental refuge addition reduced the intensity of this density dependence. Juvenile growth also declined with increasing adult goby density, but this effect was similar in both low- and high-refuge habitats. Refuge availability had no consistent effect on selective mortality, but adult goby density was significantly related to the intensity of size-selective mortality: bigger juveniles were favored where adults were abundant, and smaller juveniles were favored where adults were rare. Given the typically large difference in sizes of juveniles and adults, similar stage-structured interactions may be common but underappreciated in many marine species.


Locally Fixed Alleles: A method to localize gene drive to island populations.
Invasive species pose a major threat to biodiversity on islands. While successes have been achieved using traditional removal methods, such as toxicants aimed at rodents, these approaches have limitations and various off-target effects on island ecosystems. Gene drive technologies designed to eliminate a population provide an alternative approach, but the potential for drive-bearing individuals to escape from the target release area and impact populations elsewhere is a major concern. Here we propose the "Locally Fixed Alleles" approach as a novel means for localizing elimination by a drive to an island population that exhibits significant genetic isolation from neighboring populations. Our approach is based on the assumption that in small island populations of rodents, genetic drift will lead to alleles at multiple genomic loci becoming fixed. In contrast, multiple alleles are likely to be maintained in larger populations on mainlands. Utilizing the high degree of genetic specificity achievable using homing drives, for example based on the CRISPR/Cas9 system, our approach aims at employing one or more locally fixed alleles as the target for a gene drive on a particular island. Using mathematical modeling, we explore the feasibility of this approach and the degree of localization that can be achieved. We show that across a wide range of parameter values, escape of the drive to a neighboring population in which the target allele is not fixed will at most lead to modest transient suppression of the non-target population. While the main focus of this paper is on elimination of a rodent pest from an island, we also discuss the utility of the locally fixed allele approach for the goals of population suppression or population replacement. Our analysis also provides a threshold condition for the ability of a gene drive to invade a partially resistant population.


Optimizing protein representations with information theory.
The problem of describing a protein representation by breaking up the amino acids atoms into functionally similar atom groups has been addressed by many researchers in the past 25 years. They have used a variety of physical, chemical and biological criteria of varying degrees of rigor to essentially impose our understanding of protein structures onto various atom-typing schemes used in studies of protein folding, protein-protein and protein-ligand interactions, and others. Here, instead, we have chosen to rely primarily on the data and use information-theoretic techniques to dissect it. We show that we can obtain an optimized protein representation for a given alphabet size from protein monomers or protein interface datasets that are in agreement with general concepts of protein energetics. Closer inspection of the atom partitions led to interesting observations pointing to the greater importance of the hydrophobic interactions in protein monomers compared to interfaces and, conversely, greater importance of polar/charged interaction in protein interfaces. Comparing the atom partitions from the two datasets we show that the two are strikingly similar at alphabet size of five, proving that despite some differences, the general energetic concepts are very similar for folding and binding. Implications for further structural studies are discussed.


Scale, causes, and implications of the primary care nursing shortage.
As the demand for health care consistently rises and many individuals, even within developed countries, lack access to primary care services, a better understanding of how primary care is defined, the main causes of the primary care shortage both within the United States and across the globe, and key solutions to these issues are paramount. Upon review of the US and international primary care literature, the authors first briefly discuss the fluidity of how primary care is defined and how it is applied in nations with differing levels of health care infrastructure. The main causes of the deficit both domestically and globally are then examined. Finally, upon careful review of the research produced within the past seven years, this article suggests strategies that maximize the primary care workforce: the effective use of technology, task shifting, interprofessional teams, and more consistent primary care data to build workforce strategies.


Gain-of-function cardiomyopathic mutations in RBM20 rewire splicing regulation and re-distribute ribonucleoprotein granules within processing bodies.
Mutations in the cardiac splicing factor RBM20 lead to malignant dilated cardiomyopathy (DCM). To understand the mechanism of RBM20-associated DCM, we engineered isogenic iPSCs with DCM-associated missense mutations in RBM20 as well as RBM20 knockout (KO) iPSCs. iPSC-derived engineered heart tissues made from these cell lines recapitulate contractile dysfunction of RBM20-associated DCM and reveal greater dysfunction with missense mutations than KO. Analysis of RBM20 RNA binding by eCLIP reveals a gain-of-function preference of mutant RBM20 for 3' UTR sequences that are shared with amyotrophic lateral sclerosis (ALS) and processing-body associated RNA binding proteins (FUS, DDX6). Deep RNA sequencing reveals that the RBM20 R636S mutant has unique gene, splicing, polyadenylation and circular RNA defects that differ from RBM20 KO. Super-resolution microscopy verifies that mutant RBM20 maintains very limited nuclear localization potential; rather, the mutant protein associates with cytoplasmic processing bodies (DDX6) under basal conditions, and with stress granules (G3BP1) following acute stress. Taken together, our results highlight a pathogenic mechanism in cardiac disease through splicing-dependent and -independent pathways.


Acute onset and worsening of diabetes concurrent with administration of statins.
We report a patient in whom the administration of HMG CoA reductase inhibitors (statins) might have triggered the onset and worsening of diabetes. The patient was a 48-year-old Japanese man who underwent annual medical examination but had never been told of hyperglycemia. Four months after the commencement of atorvastatin (10 mg/day) treatment, a diagnosis of diabetes mellitus was made from his typical symptoms of hyperglycemia, postprandial plasma glucose level of 29.8 mmol/l and HbA1c of 11.5%. After 2 months of insulin therapy and 3 months after the cessation of atorvastatin, almost complete resolution of diabetes was observed. During the subsequent 3 months, diet therapy alone was sufficient to control blood glucose level. Then, we prescribed pravastatin (20 mg/day). During the subsequent 3 months, HbA1c was gradually increased. However, after discontinuation of pravastatin, HbA1c was gradually decreased. In the general population, statin does not seem to have critical adverse effects on glucose tolerance, but it may uncommonly modify the natural course of the development of diabetes in certain patients.