Single versus customized treatment planning for image-guided high-dose-rate brachytherapy for cervical cancer: dosimetric comparison and predicting factor for organs at risk overdose with single plan approach.
To compare the dose distribution between customized planning (CP) and adopting a single plan (SP) in multifractionated high-dose-rate brachytherapy and to establish predictors for the necessity of CP in a given patient. A total of 50 computed tomography-based plans for 10 patients were evaluated. Each patient had received 6 Gy for five fractions. The clinical target volume and organs at risk (i.e., rectum, bladder, sigmoid, and small bowel) were delineated on each computed tomography scan. For the SP approach, the same dwell position and time was used for all fractions. For the CP approach, the dwell position and time were reoptimized for each fraction. Applicator position variation was determined by measuring the distance between the posterior bladder wall and the tandem at the level of the vaginal fornices. The organs at risk D(2cc) (dose to 2 cc volume) was increased with the SP approach. The dose variation was statistically similar between the tandem and ring and tandem and ovoid groups. The bladder D(2cc) dose was 81.95-105.42 Gy(2) for CP and 82.11-122.49 Gy(2) for SP. In 5 of the 10 patients, the bladder would have been significantly overdosed with the SP approach. The variation of the posterior bladder wall distance from that in the first fraction was correlated with the increase in the bladder D(2cc) (SP/CP), with a correlation coefficient of -0.59. Our results support the use of CP instead of the SP approach to help avoid a significant overdose to the bladder. This is especially true for a decrease in the posterior wall distance of >/=0.5 cm compared with that in the first fraction.


Effect of low-dose aspirin on fetal and maternal generation of thromboxane by platelets in women at risk for pregnancy-induced hypertension.
There is evidence that aspirin in low doses favorably influences the course of pregnancy-induced hypertension, but the mechanism, although assumed to involve suppression of the production of thromboxane by platelets, has not been established. We performed a randomized study of the effect of the long-term daily administration of 60 mg of aspirin (n = 17) or placebo (n = 16) on platelet thromboxane A2 and vascular prostacyclin in women at risk for pregnancy-induced hypertension. Low doses of aspirin were associated with a longer pregnancy and increased weight of newborns. Serum levels of thromboxane B2, a stable product of thromboxane A2, were almost completely (greater than 90 percent) inhibited by low doses of aspirin. The urinary excretion of immunoreactive thromboxane B2 was significantly reduced without changes in the level of 6-keto-prostaglandin F1 alpha, a product of prostacyclin. Mass spectrometric analysis showed that aspirin reduced the excretion of the 2,3-dinor-thromboxane B2 metabolite--mainly of platelet origin--by 81 percent and of thromboxane B2, probably chiefly of renal origin, by 59 percent. The urinary excretion of 6-keto-prostaglandin F1 alpha and of its metabolite 2,3-dinor-6-keto-prostaglandin F1 alpha was not affected. Low doses of aspirin only partially (63 percent) reduced neonatal serum thromboxane B2. No hemorrhagic complications were observed in the newborns. Thus, in women at risk for pregnancy-induced hypertension, low doses of aspirin selectively suppressed maternal platelet thromboxane B2 while sparing vascular prostacyclin, but only partially suppressed neonatal platelet thromboxane B2, allowing hemostatic competence in the fetus and newborn.


The human interleukin-2 receptor.
Complementary DNAs corresponding to the human receptor for interleukin-2 (IL-2) have been molecularly cloned, sequenced, and expressed in both COS-1 and L cells. The human genome appears to contain a single structural gene for this receptor located on the short arm of chromosome 10 (band 14-15). However, when transcribed, at least two families of mRNAs are produced, which vary in length due to the use of at least three different polyadenylation signals. Sequence analysis of the cloned cDNAs and S1 nuclease protection assays indicate an alternative pathway of mRNA processing for this receptor whereby a 216 base-pair segment contained within the protein coding region is spliced, resulting in an mRNA unable to encode a functional IL-2 receptor. In contrast, cDNAs corresponding to mRNA retaining this 216 base-pair region code membrane receptors that bind both IL-2 and anti-Tac (monoclonal anti-IL-2 receptor antibody). Analysis of the deduced amino acid sequence reveals that the receptor is composed of 272 amino acids including a signal peptide 21 amino acids in length. Hydrophobicity analysis suggests a single, 19 amino acid transmembrane domain. A short intracytoplasmic domain composed of 13 amino acids is present and contains two potential phosphate acceptor sites (serine and threonine but not tyrosine) as well as positively charged residues presumably involved in cytoplasmic anchoring. Two sites for N-linked glycosylation sites and numerous extracytoplasmic O-linked glycosylation sites are present.


Comparative analysis of hmuO function and expression in Corynebacterium species.
We have constructed defined deletions in the hmuO gene from Corynebacterium diphtheriae and Corynebacterium ulcerans and show that the C. ulcerans hmuO mutation results in a significant reduction in hemoglobin-iron utilization, whereas in C. diphtheriae strains, deletion of hmuO caused no or only partial reduction in the utilization of heme as an iron source. We also show that expression from the C. ulcerans hmuO promoter exhibits minimal regulation by iron and heme whereas transcription from the C. diphtheriae hmuO promoter shows both significant iron repression and heme-dependent activation. These findings indicate that variability in HmuO function and expression exists among Corynebacterium species.


Evidence for active chloride accumulation in normal and denervated rat lumbrical muscle.
Intracellular Cl- activity (aiCl) was measured with Cl(-)-sensitive microelectrodes in normal and denervated rat lumbrical muscle. In normal muscle bathed in normal Krebs solution, aiCl lay close to that predicted by the Nernst equation. The addition of 9-anthracene carboxylic acid, which blocks Cl- conductance, caused aiCl to increase far above that predicted by a passive distribution. Furosemide (10 microM) reversibly blocked this accumulation. After muscle denervation, aiCl progressively increased for 1-2 wk. The rise occurred in two stages. The initial stage (1-3 d after denervation) reflected passive Cl- accumulation owing to membrane depolarization. At later times, aiCl continued to increase, with no further change in membrane potential, which suggests an active uptake mechanism. This rise approximately coincided with the natural reduction in membrane conductance to Cl- that occurs several days after denervation. Na+ replacement, K+ replacement, and furosemide each reversibly blocked the active Cl- accumulation in denervated muscle. Quantitative estimates suggested that there was little difference between Cl- flux rates in normal and denervated muscles. The results can be explained by assuming that, in normal muscle, an active accumulation mechanism operates, but that Cl- lies close to equilibrium owing to the high membrane conductance to Cl-. The rise in aiCl after denervation can be accounted for by the membrane depolarization, the reduction in membrane Cl- conductance, and the nearly unaltered action of an inwardly directed Cl- "pump."


Could Google Help Curb Online Advertising of Unhealthy Foods to U.S. Children?
In 2020, Google took voluntary action to restrict food and beverage advertising through its online channels in the European Union/United Kingdom using Google's own nutrient profiling model to identify products eligible to be marketed to children through its Google Display Network. The objective of this study was to evaluate the potential impact of the Google policy, if applied to the U.S. market, on restricting online advertising of the top-selling packaged foods and beverages in the U.S. The top 25 U.S. food and beverage manufacturers were identified. Nutrient data for products from these manufacturers were sourced from Label Insight (a Nielsen IQ company) in 2021. Each product was examined against four nutrient profiling models: the Google nutrient profiling model, the WHO Europe nutrient profiling model, the Pan American Health Organization nutrient profiling model, and the Chilean government nutrient profiling model. Under Google's nutrient profiling model, 18% of 14,188 products were eligible to be advertised to children, representing $44 billion in revenue for the top 25 U.S. manufacturers of the >$240 billion generated annually. The Google nutrient profiling model permitted the most products to be advertised to children of all four nutrient profiling models examined. U.S. children engage extensively with online media. In place of government regulation, the Google advertising policy and related nutrient profiling model would limit online advertising of the most unhealthful products to children, if the policy were to be applied to the U.S. market. The effectiveness of the policy would be strengthened by refining the Google nutrient profiling model to better align with nutrient profiling models developed by authoritative health agencies, including the WHO.


Cauda equina schwannoma presenting with intratumoral hemorrhage and intracranial subarachnoid hemorrhage.
The authors report the case of a spinal intradural schwannoma presenting with intracranial subarachnoid hemorrhage (SAH). Cerebral angiography did not show any intracranial lesion; however, MRI revealed two separate tumors in the lower segment of the spinal cord. The proximal lesion arising from the conus medullaris was well circumscribed and homogeneously enhanced, whereas the tumor in the cauda equina revealed hemorrhagic signals on MRI. This case also illustrates an unusual presentation of intracranial SAH simultaneously with intratumoral hemorrhage in a spinal cord schwannoma. The absence of hemorrhagic changes in the lesion arising proximal to the cauda equina region supports the mechanical theory proposed for the pathogenesis of hemorrhagic complications in spinal cord tumors.


Abdominal wall and chest wall reconstruction.
After studying this article, the participant should be able to: (1) Define the goals of abdominal wall and chest wall reconstruction. (2) Discuss the general principles of and the different surgical techniques for abdominal wall and chest wall reconstruction. (3) List the major advantages and disadvantages of synthetic and bioprosthetic mesh in abdominal and chest wall reconstruction. (4) Define the indications for and major factors to consider in chest wall skeletal stability reconstruction. (5) List the flaps commonly used for chest wall and abdominal wall reconstruction. Plastic surgeons commonly face reconstructive challenges in repairing the abdominal and chest walls. Reconstructive options in these two areas are rapidly expanding with the availability of new techniques and new products. The purpose of this article is to provide an updated summary of reconstruction of the abdominal and chest walls, focusing on commonly encountered problems.


Transcriptome analysis of the normal human mammary cell commitment and differentiation process.
Mature mammary epithelial cells are generated from undifferentiated precursors through a hierarchical process, but the molecular mechanisms involved, particularly in the human mammary gland, are poorly understood. To address this issue, we isolated highly purified subpopulations of primitive bipotent and committed luminal progenitor cells as well as mature luminal and myoepithelial cells from normal human mammary tissue and compared their transcriptomes obtained using three different methods. Elements unique to each subset of mammary cells were identified, and changes that accompany their differentiation in vivo were shown to be recapitulated in vitro. These include a stage-specific change in NOTCH pathway gene expression during the commitment of bipotent progenitors to the luminal lineage. Functional studies further showed NOTCH3 signaling to be critical for this differentiation event to occur in vitro. Taken together, these findings provide an initial foundation for future delineation of mechanisms that perturb primitive human mammary cell growth and differentiation.


A microfilament formation inhibitor, cytochalasin strongly enhances the low-affinity Fc epsilon receptor II (CD23) expression on the human monocyte-like cell line, U937.
Enhancement of the low-affinity Fc epsilon receptor (CD23) expression by cytochalasin was analyzed on the human monocytelike cell line, U937. The CD23 expression on the U937 cells was enhanced at 24 hr after culture with cytochalasin B, D, or E, especially cytochalasin E having the most remarkable effect on it at the low concentration. This enhanced expression was found to be associated with a concomitant increase of a CD23 (about 45-kDa) protein on the U937 cells as assessed by Western blotting analysis. On the other hand, CD11a, CD18, CD31, CD49d, or CD54 was not markedly enhanced on the U937 cells by culture with cytochalasin E, although the mean fluorescence intensities (MFIs) of CD11a, CD18, and CD54 on U937 was partially up-regulated. Cell growth of U937 cultured with cytochalasin E was completely suppressed for 72 hr, but cell viability was sufficiently maintained (more than 95%). Soluble-formed CD23 (sCD23) also was released from the U937 cells at 24 to 72 hr after culture with cytochalasin E. In addition, the protein tyrosine kinase activity was detected in the U937 cells cultured with cytochalasin E for 24 hr using the enzyme immunoassay. Enhancement of the CD23 expression on the U937 cells at 24 to 72 hr cultured with cytochalasin E was sufficiently blocked by protein tyrosine kinase inhibitors herbimycin A and genistein, and a protein synthesis inhibitor, cychloheximide. On the other hand, protein kinase C inhibitors such as H-7 and H-8 had no effect on this CD23 expression. These results suggest that a mechanism underlying enhancement of the CD23 expression on the U937 cells cultured with cytochalasin E is mediated through tyrosine phosphorylation and protein synthesis.


Hypoechoic images in hepatic steatosis: a recent ultrasound finding in differential diagnosis of space-occupying lesions of the liver.
The Authors described a particular ultrasound finding which can sometimes be observed in the echographic picture of diffused hepatic steatosis and which was seen by these Authors in 117 out of 312 patients affected by steatosis. This finding consists of the presence of one or more hypoechoic focal areas situated at the IV-V and/or II-III segment level surrounded by wide-spread increase in echogenicity of remaining parenchyma, which is typical of "bright liver". These areas, which can easily be mistaken for neoplastic formations, were instead seen to be limited areas of normal liver parenchyma free of fatty infiltration. In those patients presenting valid reasons for suspecting the presence of primary or secondary neoplastic formations, the Authors believe that echographic examination of these areas should always be accompanied by more invasive methods in order to confirm or exclude the possible presence of neoplastic formations with certainty. On the contrary, in those patients where no such diagnosis is suspected, it should prove sufficient to monitor these ares by means of ultrasound.


Inpatient characteristics of the child admitted with chronic pain.
To define the demographic, diagnostic, procedural, and episode of care characteristics for children admitted with chronic pain. We used the Pediatric Health Information System database to obtain data on demographic characteristics, length of stay, readmission rates, diagnoses, and procedures for children admitted with chronic pain. Patients with sickle cell disease, cancer, burns, cerebral palsy, transplants, and ventilator-dependent children were excluded. A total of 3752 patients with chronic pain were identified from 2004 through 2010. Admissions increased by 831% over this time period. The mean age of these patients was 13.5 years, the most common race was white (79%), and female subjects outnumbered male subjects by 2.41 to 1. The most common admission and principal discharge diagnosis was abdominal pain; comorbid diagnoses were common, with a mean of 10 diagnoses per patient. In total, 65% of patients had a comorbid gastrointestinal diagnosis and 44% had a psychiatric diagnosis. The mean length of stay was 7.32 days, with an expected length of stay of 4.24 days; 12.5% were readmitted at least once within 1 year. They underwent a mean of 3.18 procedures per patient. The average child admitted with chronic pain is a teenaged female with a wide variety of comorbid conditions, many of which are gastrointestinal and psychiatric in nature. Admissions for chronic pain are rising and account for substantial resource utilization. Future studies should further characterize this population, with the overall objective of improving outcomes and optimizing cost-effective care.


Regulation of cyclin D1 RNA stability by SNIP1.
Cyclin D1 expression represents one of the key mitogen-regulated events during the G(1) phase of the cell cycle, whereas Cyclin D1 overexpression is frequently associated with human malignancy. Here, we describe a novel mechanism regulating Cyclin D1 levels. We find that SNIP1, previously identified as a regulator of Cyclin D1 expression, does not, as previously thought, primarily function as a transcriptional coactivator for this gene. Rather, SNIP1 plays a critical role in cotranscriptional or posttranscriptional Cyclin D1 mRNA stability. Moreover, we show that the majority of nucleoplasmic SNIP1 is present within a previously undescribed complex containing SkIP, THRAP3, BCLAF1, and Pinin, all proteins with reported roles in RNA processing and transcriptional regulation. We find that this complex, which we have termed the SNIP1/SkIP-associated RNA-processing complex, is coordinately recruited to both the 3' end of the Cyclin D1 gene and Cyclin D1 RNA. Significantly, SNIP1 is required for the further recruitment of the RNA processing factor U2AF65 to both the Cyclin D1 gene and RNA. This study shows a novel mechanism regulating Cyclin D1 expression and offers new insight into the role of SNIP1 and associated proteins as regulators of proliferation and cancer.


Carotid sinus distension and superior cervical ganglion transmission.
The functional relationship between sinusal distension and the sympathetic innervation of the feline carotid sinus was investigated by neurophysiological techniques. A balloon catheter was used to distend the sinus region while recording evoked postganglionic activity from the superior cervical ganglion. When the SCG was stimulated at 1.0 or 10.0/s, balloon inflation reduced postganglionic action potential height by 25-50% and increased ganglionic negativity. These effects were subsequently abolished by section of one of the remaining intact postganglionic trunks. This evidence suggests that a pressure-modulated reflex arc, similar to the intestinointestinal reflex, might exist between the carotid sinus and superior and cervical ganglion.


FibAR: Embedding Optical Fibers in 3D Printed Objects for Active Markers in Dynamic Projection Mapping.
This paper presents a novel active marker for dynamic projection mapping (PM) that emits a temporal blinking pattern of infrared (IR) light representing its ID. We used a multi-material three dimensional (3D) printer to fabricate a projection object with optical fibers that can guide IR light from LEDs attached on the bottom of the object. The aperture of an optical fiber is typically very small; thus, it is unnoticeable to human observers under projection and can be placed on a strongly curved part of a projection surface. In addition, the working range of our system can be larger than previous marker-based methods as the blinking patterns can theoretically be recognized by a camera placed at a wide range of distances from markers. We propose an automatic marker placement algorithm to spread multiple active markers over the surface of a projection object such that its pose can be robustly estimated using captured images from arbitrary directions. We also propose an optimization framework for determining the routes of the optical fibers in such a way that collisions of the fibers can be avoided while minimizing the loss of light intensity in the fibers. Through experiments conducted using three fabricated objects containing strongly curved surfaces, we confirmed that the proposed method can achieve accurate dynamic PMs in a significantly wide working range.


Active case detection for malaria elimination: a survey among Asia Pacific countries.
Moving from malaria control to elimination requires national malaria control programmes to implement strategies to detect both symptomatic and asymptomatic cases in the community. In order to do this, malaria elimination programmes follow up malaria cases reported by health facilities to carry out case investigations that will determine the origin of the infection, whether it has been imported or is due to local malaria transmission. If necessary, the malaria programme will also carry out active surveillance to find additional malaria cases in the locality to prevent further transmission. To understand current practices and share information on malaria elimination strategies, a survey specifically addressing country policies on case investigation and reactive case detection was carried out among fourteen countries of the Asia Pacific Malaria Elimination Network (APMEN). A questionnaire was distributed to the malaria control programme managers amongst 14 countries in the Asia Pacific who have national or sub-national malaria elimination goals. Results indicate that there are a wide variety of case investigation and active case detection activities employed by the 13 countries that responded to the survey. All respondents report conducting case investigation as part of surveillance activities. More than half of these countries conduct investigations for each case. Over half aim to accomplish the investigation within one to two days of a case report. Programmes collect a broad array of demographic data during investigation procedures and definitions for imported cases are varied across respondents. Some countries report intra-national (from a different province or district) importation while others report only international importation (from a different country). Reactive case detection in respondent countries is defined as screening households within a pre-determined radius in order to identify other locally acquired infections, whether symptomatic or asymptomatic. Respondents report that reactive case detection can be triggered in different ways, in some cases with only a single case report and in others if a defined threshold of multiple cases occurs. The spatial range of screening conducted varies from a certain number of households to an entire administrative unit (e g, village). Some countries target symptomatic people whereas others target all people in order to detect asymptomatic infections. The majority of respondent programmes collect a range of information from those screened for malaria, similar to the range of information collected during case investigation. Case investigation and reactive case detection are implemented in the malaria elimination programmes in the Asia Pacific, however practices vary widely from country to country. There is little evidence available to support countries in deciding which methods to maintain, change or adopt for improved effectiveness and efficiency. The development and use of common evaluation metrics for these activities will allow malaria programmes to assess performance and results of resource-intensive surveillance measures and may benefit other countries that are considering implementing these activities.


A multispecies assessment of wildlife impacts on local community livelihoods.
Conflicts between the interests of agriculture and wildlife conservation are a major threat to biodiversity and human well-being globally. Addressing such conflicts requires a thorough understanding of the impacts associated with living alongside protected wildlife. Despite this, most studies reporting on human-wildlife impacts and the strategies used to mitigate them focus on a single species, thus oversimplifying often complex systems of human-wildlife interactions. We sought to characterize the spatiotemporal patterns of impacts by multiple co-occurring species on agricultural livelihoods in the eastern Okavango Delta Panhandle in northern Botswana through the use of a database of 3264 wildlife-incident reports recorded from 2009 to 2015 by the Department of Wildlife and National Parks. Eight species (African elephants [Loxodonta africana], hippopotamuses [Hippopotamus amphibious], lions [Panthera leo], cheetah [Acinonyx jubatus], African wild dogs [Lycaon pictus], hyenas [Crocuta crocuta], leopards [Panthera pardus], and crocodiles [Crocodylus niloticus]) appeared on incident reports, of which 56.5% were attributed to elephants. Most species were associated with only 1 type of damage (i.e., either crop damage or livestock loss). Carnivores were primarily implicated in incident reports related to livestock loss, particularly toward the end of the dry season (May-October). In contrast, herbivores were associated with crop-loss incidents during the wet season (November-April). Our results illustrate how local communities can face distinct livelihood challenges from different species at different times of the year. Such a multispecies assessment has important implications for the design of conservation interventions aimed at addressing the costs of living with wildlife and thereby mitigation of the underlying conservation conflict. Our spatiotemporal, multispecies approach is widely applicable to other regions where sustainable and long-term solutions to conservation conflicts are needed for local communities and biodiversity.


ZNF238 is expressed in postmitotic brain cells and inhibits brain tumor growth.
Brain tumors such as medulloblastoma (MB) and glioblastoma multiforme (GBM) can derive from neural precursors. For instance, many MBs are thought to arise from the uncontrolled proliferation of cerebellar granule neuron precursors (GNP). GNPs normally proliferate in early postnatal stages in mice but then they become postmitotic and differentiate into granule neurons. The proliferation of neural precursors, GNPs, as well as at least subsets of GBM and MB depends on Hedgehog signaling. However, the gene functions that are lost or suppressed in brain tumors and that normally promote the proliferation arrest and differentiation of precursors remain unclear. Here we have identified a member of the BTB-POZ and zinc finger family, ZNF238, as a factor highly expressed in postmitotic GNPs and differentiated neurons. In contrast, proliferating GNPs as well as MB and GBM express low or no ZNF238. Functionally, inhibition of ZNF238 expression in mouse GNPs decreases the expression of the neuronal differentiation markers MAP2 and NeuN and downregulates the expression of the cell cycle arrest protein p27, a regulator of GNP differentiation. Conversely, reinstating ZNF238 expression in MB and GBM cells drastically decreases their proliferation and promotes cell death. It also downregulates cyclin D1 while increasing MAP2 and p27 protein levels. Importantly, ZNF238 antagonizes MB and GBM tumor growth in vivo in xenografts. We propose that the antiproliferative functions of ZNF238 in normal GNPs and possibly other neural precursors counteract brain tumor formation. ZNF238 is thus a novel brain tumor suppressor and its reactivation in tumors could open a novel anticancer strategy.


A mechanism-based complementary screening approach for the amelioration and reversal of neurobehavioral teratogenicity.
The identification of mechanisms and outcomes for neurobehavioral teratogenesis is critical to our ability to develop therapies to ameliorate or reverse the deleterious effects of exposure to developmental neurotoxicants. We established mechanistically-based complementary models for the study of cholinergic systems in the mouse and the chick, using both environmental neurotoxicants (chlorpyrifos, perfluoroalkyls) and drugs of abuse (heroin, nicotine, PCP). Behavioral evaluations were made using the Morris maze in the mouse, evaluating visuospatial memory related to hippocampal cholinergic systems, and imprinting in the chick, examining behavior dependent on cholinergic innervation of the IMHV. In both models we demonstrated the dependence of neurobehavioral deficits on impairment of cholinergic receptor-induced expression, and translocation of specific PKC isoforms. Understanding this mechanism, we were able to reverse both the synaptic and behavioral deficits with administration of neural progenitors. We discuss the prospects for clinical application of neural progenitor therapy, emphasizing protocols for reducing or eliminating immunologic rejection, as well as minimizing invasiveness of procedures through development of intravenous administration protocols.


The experience of caring in relatives to patients with serious mental illness: gender differences, health and functioning.
The aims of the study were twofold. First, the study examined the extent to which there are gender differences in the experience of caring in a sample of relatives of patients with serious mental illness. Secondly, the study examined the association of self-reports of health and functioning with the experience of burden among relatives of patients with serious mental illness. A total of 50 relatives of 32 patients receiving psychiatric treatment for their bipolar or psychotic condition completed the Experience of Caregiving Inventory (ECI) and COOP-WONCA (a self-report measure of general state of health and functioning). The study revealed that female relatives had a higher score on eight of 10 ECI subscores, and in both ECI negative and ECI positive. Additionally, the study revealed significant correlations between a higher score on the COOP-WONCA feeling subscore and the following ECI subscores: stigma (r = 0.304, p < 0.05), problem with services (r = 0.348, p < 0.05), need to back up (r = 0.335, p < 0.05), loss (r = 0.293, p < 0.05) and the total ECI negative subscore (r = 0.343, p < 0.05). We also found a significant correlation between a higher score on the overall health subscore (COOP-WONCA) and a higher score on problems with services (r = 0.290, p < 0.05). Gender was the strongest predictor of the ECI negative subscore. Neither the COOP-WONCA subscore physical health, daily activities or social activities correlated significantly with any of the ECI subscores. Our results showed strong associations between poor mental well-being and caregiver burden. We also found a higher association with gender and the feelings subscore (COOP-WONCA) on ECI negative than we had expected.


Postnatal expression of TrkB receptor in rat vestibular nuclear neurons responsive to horizontal and vertical linear accelerations.
We examined the maturation expression profile of tyrosine kinase B (TrkB) receptor in rat vestibular nuclear neurons that were activated by sinusoidal linear acceleration along the horizontal or vertical axis. The otolithic origin of Fos expression in these neurons was confirmed with labyrinthectomized controls and normal controls, which showed only sporadically scattered Fos-labeled neurons in the vestibular nucleus. In P4-6 test rats, no Fos-labeled neurons were found in the vestibular nucleus, but the medial and spinal vestibular neurons showed weak immunoreactivity for TrkB. The intensity of TrkB immunoreactivity in vestibular nuclear neurons progressively increased in the second postnatal week but remained low in adults. From P7 onward, TrkB-expressing neurons responded to horizontal or vertical otolithic stimulation with Fos expression. The number of Fos-labeled vestibular nuclear neurons expressing TrkB increased with age, from 13-43% in P7 rats to 85-90% in adult rats. Our results therefore suggest that TrkB/neurotrophin signaling plays a dominant role in modulating vestibular nuclear neurons for the coding of gravity-related horizontal head movements and for the regulation of vestibular-related behavior during postnatal development.


When it's hard to be sure of the patient's own goals.
As the new world of managed care transforms your professional life, it also poses new ethical problems. This month, Managed Care welcomes to its pages columnist John La Puma, M.D., who will address the ethical concerns introduced by capitation and similar features of a fast-changing environment.


Determinants of enamel decalcification during simulated orthodontic treatment.
To determine the relative effects that phosphoric acid etching, metal brackets, composite resin adhesives, and filled resin sealants have on enamel decalcification. One hundred and fifty teeth were randomly divided into six groups to test the effects of acid etching, brackets, and a composite resin adhesive. The right side of each tooth was sealed. The teeth were submitted to a simulated carious challenge by Streptococcus mutans in a nutritive media for 42 days. Scanning electron photomicrographs (SEMs) were used to qualitatively evaluate the tooth surfaces. DIAGNOdent laser fluorescence was used to quantify the decalcification changes that occurred. SEMs showed normal enamel on the control teeth, etching patterns on the acid-etched teeth, surface degradation on the teeth that had resin, and sealant tags on the sealed side. The untreated control group showed significantly less decalcification (P ≤ .05) than did all groups except the no etch+bracket group. The no etch+bracket group showed significantly less decalcification than did the etch+bracket, no etch+resin, and the etch+resin groups. Acid-etched teeth demonstrated significantly more decalcification (34%) than did those that were not etched. The teeth with resin showed significantly more decalcification (24%) than did those without resin. There was no significant difference between teeth that did and did not have brackets. The unsealed side of the teeth showed small amounts of--but significantly more--decalcification (5%) compared to the sealed side. Acid etching and composite resin adhesives increase enamel decalcification in vitro, while brackets do not. Filled resin sealants protect the teeth from decalcification.


Family Life Goes On: Disability in Contemporary Families.
Disability is part of life for most contemporary families, but to date the literature on disability in families is fragmented and narrow. This editorial commentary introduces the content and findings of peer-reviewed articles appearing in a special issue of Family Relations. The editors outline unanswered but core research questions and preview the themes present in the issue: families with disabilities are diverse; economic hardship disproportionately characterizes their lives; family life with disabilities is a journey that includes stress and resilience, with support contributing significantly to the latter; and that work benefits and taxes family life. Articles extrapolate beyond findings to explore implications for family policy and practice. The editors assert that developing understanding of how disability influences families requires a more diverse and rigorous research portfolio. They further cite the need to embed disability as a variable in a range of family studies and advocate more outlets for publication.


Twenty-year longitudinal birth cohort study of individuals diagnosed with autism spectrum disorder before seven years of age.
Previous longitudinal studies have demonstrated that psychosocial outcomes for autistic adults are very limited. However, most studies are clinic-based and liable to selection bias and major methodological problems. We conducted a long-term follow-up study with 278 autistic individuals from our previous birth cohort study comprising 31,426 individuals. All participants were born in northern Yokohama between 1988 and 1996, diagnosed with autism spectrum disorder (ASD) by age seven, and followed up over 20 years. A total of 170 consented to participate in the study. Outcome measures included overall social functioning based on work, independent living, and friendships. Moreover, the time-use data concerning social participation and activities of daily living were compared with the general population. Psychosocial outcomes in adulthood (average age 25) were very good in 13.7%, good in 25.0%, fair in 31.0%, poor in 25.6%, and very poor in 4.8% of the participants. The majority participated in major life areas of and work and education (96.4%), sports (82.1%), and recreational activities and/or hobbies (98.8%). The proportion of participants who engaged in housework and self-care was comparable to that of the general population. Participants with IQ < 50 at age five had significantly worse outcomes than those with higher IQ; however, for those with IQ ≥ 50, outcomes were not significantly associated with IQ levels. Although complete independence was difficult to accomplish, many autistic adults engaged in organized community activities and housework and self-care. Time-use survey could offer a variety of data in investigating psychosocial outcomes of ASD cross-culturally.


Intragenic mutation analysis of the human epidermal growth factor receptor (EGFR) gene in malignant human oral keratinocytes.
Alteration in epidermal growth factor receptor (EGFR) expression is frequently associated with malignant transformation of epithelial tissues, including oral mucosa. This study examines the mutations in the coding region of the human EGFR gene in normal and malignant human oral keratinocytes. To examine the intragenic mutations in the human EGFR gene, a panel of normal and malignant human oral keratinocytes were examined by a nonisotopic RNase cleavage assay. Two consistent alterations were detected. First, a polymorphism, which generates a unique BsrI restriction site, was detected at position 2073. This BsrI polymorphism was present only in malignant keratinocytes. Second, Southern blot hybridization of PCR products revealed that there is a truncated EGFR mRNA (approximately 1.5-kb) in oral squamous cell carcinoma cell lines. Similar analysis in normal cell lines revealed that this truncated EGFR transcript is also present. Immunoblotting revealed the presence of this truncated form of EGFR in all keratinocyte cell lines. These data permit us to conclude that there exists a novel truncated form of EGFR in human oral keratinocytes. Furthermore, there exists a tumor-associated BsrI polymorphic site at position 2073. The potential biological relevance of the truncated receptor and the utility of the BsrI polymorphic site for diagnostic applications are currently being explored.


Monitoring athletes through self-report: factors influencing implementation.
Monitoring athletic preparation facilitates the evaluation and adjustment of practices to optimize performance outcomes. Self-report measures such as questionnaires and diaries are suggested to be a simple and cost-effective approach to monitoring an athlete's response to training, however their efficacy is dependent on how they are implemented and used. This study sought to identify the perceived factors influencing the implementation of athlete self-report measures (ASRM) in elite sport settings. Semi-structured interviews were conducted with athletes, coaches and sports science and medicine staff at a national sporting institute (n = 30). Interviewees represented 20 different sports programs and had varying experience with ASRM. Purported factors influencing the implementation of ASRM related to the measure itself (e.g., accessibility, timing of completion), and the social environment (e.g., buy-in, reinforcement). Social environmental factors included individual, inter-personal and organizational levels which is consistent with a social ecological framework. An adaptation of this framework was combined with the factors associated with the measure to illustrate the inter-relations and influence upon compliance, data accuracy and athletic outcomes. To improve implementation of ASRM and ultimately athletic outcomes, a multi-factorial and multi-level approach is needed. Key pointsEffective implementation of a self-report measure for monitoring athletes requires a multi-factorial and multi-level approach which addresses the particular measure used and the surrounding social environment.A well-designed self-report measure should obtain quality data with minimal burden on athletes and staff.A supportive social environment involves buy-in and coordination of all parties, at both an individual and organization level.


Effects of hemofiltration on serum aprotinin levels in patients undergoing cardiopulmonary bypass.
To determine the effects of hemofiltration on serum aprotinin levels during cardiopulmonary bypass (CPB) surgery. Prospective, randomized study. University of Washington Medical Center, single institution. Patients undergoing cardiac surgery without contraindications to aprotinin administration. Patients were randomized to full-Hammersmith and half-Hammersmith dosing regimens of aprotinin and were further randomized to hemofiltration or no hemofiltration. Serum aprotinin levels were studied before CPB, 60 and 120 minutes into CPB, and at the end of CPB before protamine administration. Each group experienced a decrease in serum aprotinin levels with the institution of CPB, attributable to hemodilution and redistribution of aprotinin outside of the vascular compartment. During CPB, aprotinin levels declined further, but no significant difference was observed between patients who received hemofiltration and those who did not. Hematocrit values were significantly higher at the end of CPB in the hemofiltration groups. Patients receiving half-Hammersmith dosing regimens maintained aprotinin levels throughout CPB, which have been shown to inhibit plasmin but were lower than levels previously shown to inhibit kallikrein. Hemofiltration during CPB did not significantly alter serum aprotinin levels in patients receiving half-Hammersmith and full-Hammersmith dosing regimens of aprotinin.


Decentralized Adaptive Neural Output-Feedback DSC for Switched Large-Scale Nonlinear Systems.
In this paper, for a class of switched large-scale uncertain nonlinear systems with unknown control coefficients and unmeasurable states, a switched-dynamic-surface-based decentralized adaptive neural output-feedback control approach is developed. The approach proposed extends the classical dynamic surface control (DSC) technique for nonswitched version to switched version by designing switched first-order filters, which overcomes the problem of multiple "explosion of complexity." Also, a dual common coordinates transformation of all subsystems is exploited to avoid individual coordinate transformations for subsystems that are required when applying the backstepping recursive design scheme. Nussbaum-type functions are utilized to handle the unknown control coefficients, and a switched neural network observer is constructed to estimate the unmeasurable states. Combining with the average dwell time method and backstepping and the DSC technique, decentralized adaptive neural controllers of subsystems are explicitly designed. It is proved that the approach provided can guarantee the semiglobal uniformly ultimately boundedness for all the signals in the closed-loop system under a class of switching signals with average dwell time, and the tracking errors to a small neighborhood of the origin. A two inverted pendulums system is provided to demonstrate the effectiveness of the method proposed.


A bayesian mixture model for comparative spectral count data in shotgun proteomics.
Recent developments in mass-spectrometry-based shotgun proteomics, especially methods using spectral counting, have enabled large-scale identification and differential profiling of complex proteomes. Most such proteomic studies are interested in identifying proteins, the abundance of which is different under various conditions. Several quantitative methods have recently been proposed and implemented for this purpose. Building on some techniques that are now widely accepted in the microarray literature, we developed and implemented a new method using a Bayesian model to calculate posterior probabilities of differential abundance for thousands of proteins in a given experiment simultaneously. Our Bayesian model is shown to deliver uniformly superior performance when compared with several existing methods.


Optimizing Diffusion-Tensor Imaging Acquisition for Spinal Cord Assessment: Physical Basis and Technical Adjustments.
Diffusion-tensor imaging (DTI) has been used in the assessment of the central nervous system for the past 3 decades and has demonstrated great utility for the functional assessment of normal and pathologic white matter. Recent technical advances have permitted the expansion of DTI applications to the spinal cord. MRI of the spinal cord has traditionally been limited to conventional sequences, which provide information regarding changes in the anatomic shape of a structure or its signal intensity, suggesting the presence of a pathologic entity. However, conventional MRI lacks the ability to provide pathophysiologic information. DTI of the spinal cord can deliver pathophysiologic information on a molecular basis and thereby has several adjunctive uses. These advantages have yet to be fully evaluated, and therefore spinal DTI lacks widespread adoption. The barriers to implementation include a lack of understanding of the underlying physics principles needed to make necessary technical adjustments to obtain diagnostic images, as well as the need for standardization of protocols and postprocessing methods. The authors provide a comprehensive review of the physics of spinal cord DTI and the technical adjustments required to obtain diagnostic images and describe tips and tricks for accurate postprocessing. The primary clinical applications for spinal cord DTI are reviewed. Online supplemental material is available for this article. ©RSNA, 2020 See discussion on this article by Smith.


Optimal diffusion-weighted imaging protocol for lesion detection in transient global amnesia.
Diffusion-weighted imaging (DWI) can depict small punctate hyperintense lesions in the hippocampus in transient global amnesia (TGA). The purpose of this study was to find an optimal DWI protocol for lesion detection in TGA by investigating various imaging parameters and imaging timing after symptom onset. Sixteen patients with TGA diagnosed during 14 months underwent DWI within 24 hours and again at follow-up 3 days after onset. Each DWI session included 4 different sequences using different b-values (seconds per square millimeter) and section thicknesses (millimeter): 1000/5, 1000/3, 2000/3, and 3000/3. The presence or absence of hyperintense lesions on the 8 DWIs was determined visually, and the number of lesions detected was compared. Thirteen of the 16 patients (81%) had either single or multiple punctate hyperintense lesions, totaling 24 lesions, and the remaining 3 patients had no lesions. All lesions detected were in the hippocampus except 1. The number of lesions detected on initial DWIs at a b-value/section thickness of 1000/5, 1000/3, 2000/3, and 3000/3 was 3, 9, 13, and 13, respectively, whereas that of follow-up DWIs was 17, 22, 24, and 24, respectively. On the basis of these preliminary results, the highest lesion detection was achieved for DWI with b = 2000/3 mm or b = 3000/3 mm at 3 days postonset. When no lesion is detected by DWI within 24 hours after onset, follow-up DWI is recommended several days later.


Receptor-mediated choreography of life and death.
The cytokine tumor necrosis factor was originally identified as a protein that kills tumor cells. So far, 18 distinct members of this family have been identified. All of them regulate cell survival, proliferation, differentiation, and cell death, also called apoptosis. The apoptosis induced by TNF, and other members of the family, for example, FasL, VEGI, and TRAIL is mediated through death receptors. The apoptotic signals by these cytokines are transduced by eight different death domain- (DD) containing receptors (TNFR1, also called DR1; Fas, also called DR2; DR3, DR4, DR5, DR6, NGFR, and EDAR). The intracellular portion of all these receptors contains a region approximately 80 amino acids long referred to as the "death domain." Upon activation by its ligand, the DD recruits various proteins that mediate both death and proliferation of the cells. These proteins in turn recruit other proteins via their DDs or death effector domains. The actual destruction of the cell, however, is accomplished by serial activation of a family of proteases referred to as caspases. Cell death is negatively regulated by a family of proteins that includes decoy receptors, silencer of DD, sentrin, cellular FLICE inhibitory protein, cellular inhibitors of apoptosis, and survivin. This review is an attempt to describe how these negative and positive players of cell death perform a harmonious dance with each other.


Functional Limitations Mediate the Relationship Between Pain and Depressive Symptoms in Former NFL Athletes.
The objective of this study was to analyze data from the National Football League Player Care Foundation Study of Retired NFL Players to understand potential risks for depressive symptoms in former athletes by investigating the relationship between pain and depressive symptoms in a multivariate context, while simultaneously exploring the potential connection with functional limitations. Descriptive statistics were used to describe the study sample and to conduct bivariate comparisons by race and age cohort. Linear regression models were conducted in the subsample of respondents reporting on depressive symptoms using the PHQ-9. Models examine the relationship of bodily pain, injury as a reason for retirement or not re-signing with a team, length of NFL career, sociodemographic characteristics, chronic conditions, and functional limitations to depression. Interaction terms tested whether race and age moderated the effect of bodily pain and functional limitations on depressive symptoms. Bivariate associations revealed no significant differences between younger and older former players in indicators of pain and only slightly higher functional limitations among younger former players. In the multivariate models, pain was significantly associated with depressive symptoms (β = 0.36; p < .01), net of a range of relevant controls. Adding an index of functional limitations reduced this association by nearly half (β = 0.20; p < .01) and functional limitations was significantly associated with depressive symptoms (β = 0.40; p < .01). No statistically significant interactions were found. Overall, bodily pain was strongly associated with depressive symptoms. After accounting for the effects of functional limitations, this association was notably reduced. These results may be useful in identifying aging-related physical declines in relatively younger adult men who may be at the greatest risk for depression. They highlight how physical functionality and activity may mitigate the risk of depression, even in the presence of significant bodily pain.


[Structure of the complex receptive field of the visual cortex in the cat].
This most common type of a complex receptive field was analyzed whose response contains modulated and unmodulated components. Application of the mask covering a half of the field increased (according to the filter theory) the bandwidth of the field as a spatial frequency filter due to appearance or enhancement of the response at lateral low and high frequencies. In this case the modulated components of response from each half of the field are out of phase. Analysis of this fact together with responses to thin light and dark bars permitted describing the structure of the field as consisting of linear and nonlinear subsystems converging on the complex field neuron. The linear system comprises several pairs of on- and off-subfields of corpus geniculatum. On- and off-subfields in the pair are spatially overlapped and converge on a neuron of the linear subsystem with opposite signs. The nonlinear system comprises only on- or off-subfields. Other types of complex fields are formed by various combinations of subsystems. The results evidence that the complex field is a grating spatial frequency filter.


miRNA-21 sensitizes gastrointestinal stromal tumors (GISTs) cells to Imatinib via targeting B-cell lymphoma 2 (Bcl-2).
miRNA-21 (miRNA-21) has recently been recognized to tumor suppressive in various types of cancers. However, the role of miRNA-21 in gastrointestinal stromal tumors (GISTs) is still ambiguous. In this study, we investigated the regulation by miRNA-21 on the sensitivity of gastrointestinal stromal tumors (GISTs) cells to Imatinib. We examined the expression of miRNA-21 and B-cell lymphoma 2 (Bcl-2) in GIST specimens by the real-time quantitative PCR assay (RT-qPCR). Then we explored the regulation by miRNA-21 on the Bcl-2 expression by the RT-qPCR assay, Western blotting assay and the luciferase assay in GIST-T1 cells. In addition, we examined the influence of miRNA-21 on the sensitivity to Imatinib of GIST-T1 cells with colony forming assay and apoptotic assay. Results indicated that miRNA-21 expression was suppressed in GIST tissues. And we identified putative miRNA-21 binding sites within the 3'-untranslated region (3'-UTR) of the human Bcl-2 gene. Transient transfection of miRNA-21 mimics into human GIST GIST-T1 cell line significantly downregulated the Bcl-2 expression in both mRNA and protein levels. Moreover, the miRNA-21 mimics transfection markedly aggravated the Imatinib-mediated growth inhibition and apoptosis induction in GIST-T1 cells. Our results demonstrated that miRNA-21 suppressed Bcl-2 expression in GIST cells and could function as a potent tumor suppressor in GIST. And the miRNA-21 promotion could sensitize GIST cells to Imatinib. It implies a potential role in the GIST treatment.


Investigation of Non-covalent Interactions of Metal Complexes with DNA in Cell-free Systems.
Non-covalent interactions of metallo compounds with DNA range from the simple, unspecific electrostatic binding of a positively charged metal complex to the sequence-selective recognition of DNA binding sites due to shape, size, symmetry and hydrogen bonding complementarity of a rationally designed system. Metal complexes that recognize and target specific DNA sequences or particular structures are of considerable interest as therapeutics, diagnostics or structural probes. To gain molecular level insight into DNA-metal complex interactions, binding studies are carried out in cell-free systems using isolated DNA or short oligonucleotides. For this, a powerful toolbox of complementary spectroscopic and biophysical techniques is available. This review focuses on the most frequently applied spectroscopic methods; UV/Vis, CD, LD, fluorescence emission and NMR spectroscopy and is aimed at giving the reader an overview of the qualitative and/or quantitative information that can be obtained. After a short introduction into DNA structures and non-covalent metal complex-DNA interactions, each spectroscopic method will be discussed. In the last section a few selected studies will be described as illustrative examples for the potential of the various spectroscopic methods.


The effects of curcumin-piperine supplementation on inflammatory, oxidative stress and metabolic indices in patients with ischemic stroke in the rehabilitation phase: a randomized controlled trial.
Stroke is a leading cause of death worldwide, which is associated with a heavy economic and social burden. The purpose of this study was to investigate the effects of supplementation with curcumin-piperine combination in patients with ischemic stroke in the rehabilitation stage. In this randomized controlled trial, 66 patients with stroke were randomized into two groups receiving curcumin-piperine tablets (500 mg curcumin + 5 mg piperine) and matched placebo tablets for 12 weeks. High-sensitivity C-reactive protein (hs-CRP), carotid intima-media thickness (CIMT), thrombosis, total antioxidant capacity (TAC), lipid profile, anthropometric indices, blood pressure, and quality of life were assessed before and after the intervention. Statistical data analysis was done using SPSS22 software. A total of 56 patients with a mean age of 59.80 ± 4.25 years completed the trial. Based on ANCOVA test, adjusted for baseline values, curcumin-piperine supplementation for 12 weeks resulted in significant reductions in serum levels of hs-CRP (p = 0.026), total cholesterol (TC) (p = 0.009), triglycerides (TG) (p = 0.001), CIMT (p = 0.002), weight (P = 0.001), waist circumference (p = 0.024), and systolic and diastolic blood pressure (p < 0.001), and a significant increase in TAC (p < 0.001) in comparison to the placebo. Pain score significantly increased in both groups; however, its increase was significantly higher in the placebo group compared with the intervention group (p = 0.007). No significant changes were observed between the two groups in terms of serum fibrinogen, low-density lipoprotein (LDL), high-density lipoprotein (HDL), and quality of life indices. Curcumin-piperine supplementation had beneficial effects on CIMT, serum hs-CRP, TC, TG, TAC, and systolic and diastolic blood pressure in patients with ischemic stroke in the rehabilitation stage.


Public health in an era of personal health records: opportunities for innovation and new partnerships.
In the near future, citizens will be able to control and manage their own health information through electronic personal health record systems and tools. The clinical benefits of this innovation, such as cost savings, error reduction, and improved communication, have been discussed in the literature and public forums, as have issues related to privacy and confidentiality. Receiving little attention are the benefits these will have for public health. The benefits and potential for innovation are broad and speak directly to core public health functions such as health monitoring, outbreak management, empowerment, linking to services, and research. Coupled with this is a new relationship with citizens as key partners in protecting and promoting the public's health.


Changes in thickness and surface area of the human cortex and their relationship with intelligence.
Changes in cortical thickness over time have been related to intelligence, but whether changes in cortical surface area are related to general cognitive functioning is unknown. We therefore examined the relationship between intelligence quotient (IQ) and changes in cortical thickness and surface over time in 504 healthy subjects. At 10 years of age, more intelligent children have a slightly thinner cortex than children with a lower IQ. This relationship becomes more pronounced with increasing age: with higher IQ, a faster thinning of the cortex is found over time. In the more intelligent young adults, this relationship reverses so that by the age of 42 a thicker cortex is associated with higher intelligence. In contrast, cortical surface is larger in more intelligent children at the age of 10. The cortical surface is still expanding, reaching its maximum area during adolescence. With higher IQ, cortical expansion is completed at a younger age; and once completed, surface area decreases at a higher rate. These findings suggest that intelligence may be more related to the magnitude and timing of changes in brain structure during development than to brain structure per se, and that the cortex is never completed but shows continuing intelligence-dependent development.


Bee pollination outperforms pesticides for oilseed crop production and profitability.
Nature-based agriculture that reduces dependency on chemical inputs requires using ecological principles for sustainable agro-ecosystems, aiming to balance ecology, economics and social justice. There is growing evidence that pollinator-dependent crops with high insect, particularly bee, pollination service can give higher yields. However, the interacting effects between insect pollination and agricultural inputs on crop yields and farm economics remain to be established to reconcile food production with biodiversity conservation. We quantified individual and combined effects of pesticides, insect pollination and soil quality on oilseed rape (Brassica napus L.) yield and gross margin, using a total of 294 farmers' fields surveyed between 2013 and 2016. We show that yield and gross margins are greater (15-40%) in fields with higher pollinator abundance than in fields with reduced pollinator abundance. This effect is, however, strongly reduced by pesticide use. Greater yields may be achieved by either increasing agrochemicals or increasing bee abundance, but crop economic returns were only increased by the latter, because pesticides did not increase yields while their costs reduced gross margins.


Extra-articular Manifestations of Chikungunya.
Chikungunya fever (CHIK) is a neglected tropical disease associated with chronic arthritis. CHIK is usually a self-limiting condition; however, extra-articular manifestations present as atypical illness in a minority of patients. These atypical features may mimic other conditions and potentially distract physicians from the true diagnosis. This review analyzes the evidence of many unusual extra-articular manifestations reported in cases of CHIK. Depending on the affected system, these unusual manifestations include encephalitis, myocarditis, acute interstitial nephritis, cutaneous manifestations, acute anterior uveitis, abdominal pain, and depression. In addition, coinfections and comorbidities may cause atypical illness and obscure the diagnosis. Further studies are required to clarify the pathophysiology and natural history of CHIK, as it remains a burdening condition. Exploring its atypical symptoms may be the missing scientific piece of this puzzle.


Development of a Predictive Multiple Reaction Monitoring (MRM) Model for High-Throughput ADME Analyses Using Learning-to-Rank (LTR) Techniques.
Multiple Reaction Monitoring (MRM) is an important MS/MS technique commonly used in drug discovery and development, allowing for the selective and sensitive quantification of compounds in complex matrices. However, compound optimization can be resource intensive and requires experimental determination of product ions for each compound. In this study, we developed a Learning-to-Rank (LTR) model to predict the product ions directly from compound structures, eliminating the requirement for MRM optimization experiments. Experimentally determined MRM conditions for 5757 compounds were used to develop the model. Using the MassChemSite software, theoretical fragments and their mass-to-charge ratios were generated, which were then matched to the experimental product ions to create a data set. Each possible fragment was ranked based on its intensity in the experimental data. Different LTR models were built on a training split. Hyperparameter selection was performed using 5-fold cross validation. The models were evaluated using the Normalized Discounted Cumulative Gain at top k (NDCG@k) and the Coverage at top k (Coverage@k) metrics. Finally, the model was applied to predict MRM conditions for a prospective set of 235 compounds in high-throughput Caco-2 permeability and metabolic stability assays, and quantification results were compared to those obtained with experimentally acquired MRM conditions. The LTR model achieved a NDCG@5 of 0.732 and Coverage@5 of 0.841 on the validation split, and its predictions led to 97% of biologically equivalent results in the Caco-2 permeability and metabolic stability assays.


Automated nucleic acid amplification testing in blood banks: An additional layer of blood safety.
A total of 30 million blood components are transfused each year in India. Blood safety thus becomes a top priority, especially with a population of around 1.23 billion and a high prevalence rate of human immunodeficiency virus (HIV), hepatitis B virus (HBV) and hepatitis C virus (HCV) in general population. Nucleic acid amplification testing (NAT) in blood donor screening has been implemented in many developed countries to reduce the risk of transfusion-transmitted viral infections (TTIs). NAT takes care of the dynamics of window period of viruses and offers the safest blood pack for donation. The aim of this study is to show the value of NAT in blood screening. Dhanavantari Blood Bank, Rajahmundry, Andhra Pradesh, India. Over a period of 1 year from January 2012 to December 2012, a total number of 15,000 blood donor samples were subjected to tests for HIV, HBV, and HCV by enzyme-linked immunosorbent assay (ELISA) method and 8000 ELISA nonreactive samples were subjected for NAT using multiplex polymerase chain reaction technology. Of the 15,000 donors tested, 525 were seroreactive. In 8000 ELISA negative blood samples subjected to NAT, 4 donor samples were reactive for HBV. The NAT yield was 1 in 2000. NAT could detect HIV, HBV, and HCV cases in blood donor samples those were undetected by serological tests. NAT could interdict 2500 infectious donations among our approximate 5 million annual blood donations.


Purification and characterisation of 3-hydroxyphenylacetate 6-hydroxylase: a novel FAD-dependent monooxygenase from a Flavobacterium species.
3-Hydroxyphenylacetate 6-hydroxylase was purified 70-fold from a Flavobacterium sp. grown upon phenylacetic acid as its sole carbon and energy source. The presence of FAD and dithiothreitol during purification is essential for high recovery of active enzyme. SDS/PAGE of purified enzyme reveals a single band with a minimum molecular mass of 63 kDa. Analytical gel-filtration, sedimentation-equilibrium and sedimentation-velocity experiments indicate that the purified enzyme exists in solution mainly as a dimer, containing 1 molecule non-covalently bound FAD/subunit. 3-Hydroxyphenylacetate 6-hydroxylase utilizes NADH and NADPH as external electron donors with similar efficiency. The enzyme shows a narrow substrate specificity. Only the primary substrate 3-hydroxyphenylacetate is hydroxylated efficiently, yielding 2,5-dihydroxyphenylacetate as a product. During turnover, the substrate analogues 3,4-dihydroxyphenylacetate and 4-hydroxyphenylacetate are partially hydroxylated, exclusively at the 6' (2') position. The physiological product 2,5-dihydroxyphenylacetate acts as an effector, strongly stimulating NAD(P)H oxidation. The activity of 3-hydroxyphenylacetate 6-hydroxylase is severely inhibited by chloride ions, competitive to the aromatic substrate. In the native state of enzyme, two sulfhydryl groups are accessible to 5,5'-dithiobis(2-nitrobenzoate). Titration with stoichiometric amounts of either 5,5'-dithiobis(2-nitrobenzoate) or mercurial reagents completely blocks enzyme activity. Inactivation by cysteine reagents is inhibited by the substrate 3-hydroxyphenylacetate. The original activity is fully restored by treatment of the modified enzyme with dithiothreitol. The N-terminal amino acid sequence of the enzyme lacks the consensus sequence GXGXXG, found at the N-termini of all flavin-dependent external monooxygenases sequenced so far. The amino acid composition of 3-hydroxyphenylacetate 6-hydroxylase is also presented.


Bone induction in implants of decalcified bone and dentine.
The fate of decalcified bone and dentine implanted in muscle and beneath the kidney capsule has been studied in young rats. Quantitatively speaking there was a great deal of variation, but in general the implants became surrounded and invaded by young vascular connective tissue; then tunnels were eroded and cavities enlarged by multi-nucleated giant cells; then the matrix around erosion chambers became recalcified; and finally new bone was induced on the eroded recalcified surfaces. Erosion was much more extensive, and bone was much more readily induced in the intramuscular than in the subcapsular implants. It is concluded that the presence of an eroded, recalcified surface is a pre-requisite for bone induction under these conditions.


MRI-defined subcortical ischemic vascular disease: baseline clinical and neuropsychological findings. The LADIS Study.
Subcortical ischemic vascular disease (SIVD) is a common, but often overlooked cause of vascular cognitive impairment. Diagnostic research criteria for SIVD are based on magnetic resonance imaging (MRI) findings including substantial white matter lesions (WML) and multiple lacunar infarcts. Empirical studies validating these imaging criteria are still few. The purpose of the study was to describe the clinical and cognitive characteristics of the MRI-defined SIVD in a mixed sample of functionally independent elderly subjects with WML. The subjects of the Leukoaraiosis and Disability (LADIS) study, aged 65-84 years, underwent comprehensive clinical and neuropsychological examinations, and brain MRI at the baseline assessment. The subjects meeting the SIVD imaging criteria (n = 89) were compared to the other subjects of the sample (n = 524). SIVD was associated with lower education, hypertension and, independently, with obesity. The subjects with SIVD had more often motor impairment, a history of falls, and subtle impairment in activities of daily living, but they did not differ for depressive symptoms. SIVD subjects performed significantly inferiorly in tests of global cognitive function, psychomotor speed, attention and executive functions, verbal fluency, and working memory. In this population of nondisabled older adults with WML, SIVD was related to specific clinical and functional characteristics. Neuropsychological features included psychomotor slowing as well as deficits in attention and executive functions.


Major anatomical variations of the tracheobronchial tree: bronchoscopic observation.
Fiberoptic and rigid bronchoscopy are widely used diagnostic and therapeutic tools in pulmonary medicine. Investigators often neglect the bronchial variations; however, bronchial variations may have important implications for bronchoscopy, brachytherapy, pulmonary resections and intubations. It is accepted that anatomic variations of the airways are due to anomalies in the development of the lungs. As a result, lung buds grow to an inappropriate number or arise at atypical sites. In the present study, we tried to determine the incidence of bronchial variations in our region. We investigated 2550 consecutive reports of bronchoscopy retrospectively. Major variations of the tracheobronchial tree were found in 2.6% of patients examined by bronchoscopy. The most frequent finding was a bifurcate pattern in the right upper lobe (47.7%). The variations were localized to the right upper lobe in 71.6% of patients. Male predominance was observed in all anatomic variations except one.


[Quantitative characteristics of the frequency of chromosomal aberrations in a group of residents of a large western Siberian industrial region].
The results of a 15-year study of chromosome aberration frequency in cultured peripheral blood lymphocytes from subjects living in the Kuzbass industrial region are presented. The database for the analysis of the main parameters of chromosome aberrations contains data on 925 subjects, with the total number of cells examined being 92,900. It has been found that the total frequency of aberrant metaphases in the database is 3.73 +/- 0.1%, whereas this frequency for the sample of subjects from industrial areas of this region (the basic control group) is significantly lower (2.86 +/- 0.26%). It has been demonstrated that the sex and age of the subjects do not affect substantially the frequency of any type of chromosome aberrations. Tobacco smoking is associated with a slight increase in chromosome damage frequency; however, the difference between smokers and nonsmokers is insignificant even among subjects exposed to occupational hazards. The possible causes that have determined the increased basic and background chromosome aberration frequencies in the population of the Kemerovo oblast compared to these parameters for the European part of Russia and the CIS are discussed.


FGF16 promotes invasive behavior of SKOV-3 ovarian cancer cells through activation of mitogen-activated protein kinase (MAPK) signaling pathway.
Uncontrolled cell growth and tissue invasion define the characteristic features of cancer. Several growth factors regulate these processes by inducing specific signaling pathways. We show that FGF16, a novel factor, is expressed in human ovary, and its expression is markedly increased in ovarian tumors. This finding indicated possible involvement of FGF16 in ovarian cancer progression. We observed that FGF16 stimulates the proliferation of human ovarian adenocarcinoma cells, SKOV-3 and OAW-42. Furthermore, through the activation of FGF receptor-mediated intracellular MAPK pathway, FGF16 regulates the expression of MMP2, MMP9, SNAI1, and CDH1 and thus facilitates cellular invasion. Inhibition of FGFR as well as MAPK pathway reduces the proliferative and invasive behavior of ovarian cancer cells. Moreover, ovarian tumors with up-regulated PITX2 expression also showed activation of Wnt/β-catenin pathway that prompted us to investigate possible interaction among FGF16, PITX2, and Wnt pathway. We identified that PITX2 homeodomain transcription factor interacts with and regulates FGF16 expression. Furthermore, activation of the Wnt/β-catenin pathway induces FGF16 expression. Moreover, FGF16 promoter possesses the binding elements of PITX2 as well as T-cell factor (Wnt-responsive), in close proximity, where PITX2 and β-catenin binds to and synergistically activates the same. A detail study showed that both PITX2 and T-cell factor elements and the interaction with their binding partners are necessary for target gene expression. Taken together, our findings indicate that FGF16 in conjunction with Wnt pathway contributes to the cancer phenotype of ovarian cells and suggests that modulation of its expression in ovarian cells might be a promising therapeutic strategy for the treatment of invasive ovarian cancers.


The value of serum ischemia-modified albumin for assessing liver function in patients with chronic liver disease.
Ischemia-modified albumin (IMA), measured by the cobalt-binding capacity of albumin, is a promising biomarker for cardiac ischemia. The IMA-to-serum albumin ratio (IMAR) has been reported to relate to the severity of decompensated liver cirrhosis. This study aimed to assess IMA and IMAR as a liver function test and to investigate whether albumin infusion changes IMAR in patients with liver cirrhosis. Blood samples were collected from healthy volunteers (n=51) and patients with chronic hepatitis (n=25), liver cirrhosis (n=24) and uremia (n=13). Parameters examined included serum levels of IMA, albumin, total bilirubin, creatinine, international normalized ratio (INR), model for end-stage liver disease (MELD) score, child-turcotte-pugh (CTP) score, indocyanine green (ICG) retention rate and total antioxidant capacity (TAC). Paired serum samples from patients pre- and post-albumin infusion (n=9) were collected and the changes were compared. IMA and IMAR increased in patients with chronic hepatitis or cirrhosis, as compared to healthy volunteers. In patients with liver disease, IMA and IMAR were significantly associated with ICG retention, bilirubin, TAC and INR. In addition, IMAR was associated with CTP and MELD score in patients with cirrhosis. Albumin therapy improved patients' serum levels of creatinine and bilirubin and MELD score, but not IMA and IMAR. IMAR, reflecting liver function and oxidative stress, is a more objective liver function test as it was not affected after a 3-day albumin infusion. More investigations, however, are needed to validate the use of IMAR in cases of chronic liver disease.


High-affinity, sodium-dependent gamma-aminobutyric acid uptake by slices of rat ovary.
The high concentration of gamma-aminobutyric acid (GABA) recently demonstrated in rat ovary prompted us to examine the capacity of ovarian slices to take up [3H]GABA. Active uptake, dependent on temperature and sodium concentration, was observed and a kinetic constant (Km) of 1.0 microM found for the uptake process. Ouabain (100 microM) reduced the rate of accumulation of [3H]GABA. Uptake was inhibited only partially by 100 microM d,l-nipecotic acid, but more strongly by 100 microM beta-alanine. These results suggest that the uptake system in ovary possesses properties similar to those of high-affinity GABA transport systems in the brain.


The 50 distal amino acids of the 2AHP homing protein of Grapevine fanleaf virus elicit a hypersensitive reaction on Nicotiana occidentalis.
Avirulence factors are critical for the arm's race between a virus and its host in determining incompatible reactions. The response of plants to viruses from the genus Nepovirus in the family Secoviridae, including Grapevine fanleaf virus (GFLV), is well characterized, although the nature and characteristics of the viral avirulence factor remain elusive. By using infectious clones of GFLV strains F13 and GHu in a reverse genetics approach with wild-type, assortant and chimeric viruses, the determinant of necrotic lesions caused by GFLV-F13 on inoculated leaves of Nicotiana occidentalis was mapped to the RNA2-encoded protein 2AHP , particularly to its 50 C-terminal amino acids. The necrotic response showed hallmark characteristics of a genuine hypersensitive reaction, such as the accumulation of phytoalexins, reactive oxygen species, pathogenesis-related protein 1c and hypersensitivity-related (hsr) 203J transcripts. Transient expression of the GFLV-F13 protein 2AHP fused to an enhanced green fluorescent protein (EGFP) tag in N. occidentalis by agroinfiltration was sufficient to elicit a hypersensitive reaction. In addition, the GFLV-F13 avirulence factor, when introduced in GFLV-GHu, which causes a compatible reaction on N. occidentalis, elicited necrosis and partially restricted the virus. This is the first identification of a nepovirus avirulence factor that is responsible for a hypersensitive reaction in both the context of virus infection and transient expression.


Case report of a familial triple: a syndrome and review of the literature.
Triple-A syndrome, or Allgrove syndrome (AS), is a rare autosomal recessive disorder characterized by the alacrimia, achalasia, and adrenal insufficiency triad. Alacrimia usually starts at early infancy, while achalasia and adrenal insufficiency appear later during childhood or adulthood. Some patients may also present with the so-called Double-A syndrome (i.e., alacrimia and achalasia, or alacrimia and adrenal insufficiency); adrenal insufficiency usually represents a life-threatening event due to severe hypoglycemia. Many patients may also present other associated manifestations, such as neurological disorders. We describe, here, 2 sisters of non-consanguineous parents. An 8-year-old girl was admitted to the Pediatric Care Unit of Parma after an episode characterized by seizure with loss of consciousness and generalized hypertonia lasting a few minutes. Her sister, a 6-year-old girl, presented with recurrent episodes of vomiting and failure to thrive. Both children were investigated by laboratory tests, esophagogastroduodenoscopy, and imaging. The first patient had the complete triad of AS (alacrimia, achalasia, adrenal insufficiency), while the second one presented only alacrimia and achalasia. Both resulted from a mutation in the achalasia, addisonianism, alacrimia syndrome gene. Both patients were treated with oral hydrocortisone for Addison disease, and with artificial tears in the first case. After many pneumatic endoscopic dilations and therapy with nifedipine, both patients underwent surgical Heller myotomy for achalasia. A rapid and favorable recovery to normal diet and with improvement of growth parameters was obtained. These cases are also compared with the literature data, reported in a brief review. AS is a rare multisystemic disorder. The longer diagnosis is delayed, the greater extent to which this syndrome may be life-threatening, mainly because of hypoglycemia due to adrenal insufficiency. In AS, the red-flag symptom of alacrimia should instigate investigation for achalasia, Addison disease, and achalasia, addisonianism, alacrimia syndrome gene mutation.


The role of magnetic resonance imaging in the prenatal diagnosis of cleft lip and palate.
Cleft lip and/or palate is the most common congenital craniofacial anomaly. Ultrasonography plays a key role in the early diagnosis of this anomaly and is completed by MRI. The purpose of this paper is to present and summarize the experience in diagnosis of cleft lip/palate by means of MRI. The material consists of 62 fetuses that required more detailed evaluation which was conducted with the use of a 1.5 T scanner in SSFSE/T2 sequence in the sagittal, transverse and coronal plane. The cleft was diagnosed in 15 fetuses: an isolated cleft lip in one case (6.7%), a cleft lip and alveolar process in 2 (13.3%), a cleft lip and palate in 12 (80%). In eight fetuses (53.3%) the defect was unilateral, in 6 (40%) on both sides, in one case (6.7%) - a bilateral cleft lip and unilateral cleft palate was diagnosed. In three cases (20%), the cleft lip and/or palate defect was isolated, in 12 (80%) - it coexisted with other fetal abnormalities. MRI was less useful than ultrasound in 1 case (6.7%), in 4 cases (26.7%) it did not add any significant new information, in the remaining 10 cases (66.6%) important additional information was obtained on MRI. MRI revealed more details of the cleft in 5 cases (33.3%). In 10 fetuses (66.7%), cleft diagnosis was based on ultrasound and MRI only confirmed it. In 47 cases MRI allowed to show normal fetal faces, while there were difficulties of visualisation on ultrasound. Prenatal MRI is a method supporting fetal ultrasound and is used to confirm/expand sonographic diagnosis, but can also change it. In the case of cleft lip and palate fetal MRI produces a better picture of the connections between the cavities, the degree of involvement of the secondary palate and cleft extent, and also helps to detect/assess other associated fetal abnormalities.


A New Species of Predatory Nudibranch (Gastropoda: Trinchesiidae) of the Coral Pavona decussata.
Some nudibranchs are predators of scleractinian corals, but little is known about their diversity. Here we describe Phestilla fuscostriata sp. nov., the first species of nudibranch that preys on Pavona decussata, a structure-forming agariciid species in the South China Sea. This new species has a white body with brown pigmentation on the dorsum and cerata, and exhibits excellent mimicry by matching the colour of its coral host. The nudibranch lays crescent-shaped egg masses on the coral surface, where the embryos develop and hatch in 2-3 weeks. This new species possesses a large number of cerata that are arranged in widely-spaced rows, with each row having one dorsal ceras and zero to several ventral cerata, which distinguishes it from all other congeneric species. A comparison of the mitochondrial COI and 16S rRNA genes and the nuclear H3 gene between P. fuscostriata sp. nov. and other Phestilla spp. found that their interspecific distances are large enough to justify the recognition of the new species.


Factor analysis as a tool for survey analysis using a professional role orientation inventory as an example.
The purpose of this article is to illustrate how confirmatory factor analysis can be used to extend and clarify a researcher's insight into a survey instrument beyond that afforded through the typical exploratory factor analytic approach. The authors use as an example a survey instrument developed to measure individual differences in professional role orientation among physical therapists, the Professional Role Orientation Inventory for Physical Therapists (PROI-PT). Five hundred three physical therapists responded to a mail survey instrument that was sent to a random sample of 2,000 American Physical Therapy Association members. An adapted version of the Professional Role Orientation Inventory, a 40-item Likert-scale instrument developed to assess professional role orientation on 4 dimensions (authority, responsibility, agency, and autonomy), was used. Exploratory and confirmatory factor analyses were used to examine the factorial validity of the PROI-PT. Exploratory factor analysis served as a starting point for examining the factor structure of the instrument. Confirmatory factor analysis then was used to test the hypothesized factor structure and to suggest refinements to the PROI-PT that would improve a psychometric property (internal consistency). Although further refinement of the PROI-PT is needed, an instrument that yields valid and reliable measurements of individual differences in professionalism among physical therapists could further our understanding of the psychosocial aspects of physical therapist practice. Exploratory and confirmatory factor analyses can be used by researchers who study various psychosocial constructs in physical therapy.


Demonstration of fibronectin as a major extracellular protein of human gingival fibroblasts.
A major extracellular protein of cultured adult human gingival fibroblasts was shown to be fibronectin by indirect immunofluorescence, lactoperoxidase catalyzed iodination and metabolic radiolabeling. Fibronectin preparations from these cells possessed the biological properties of agglutination of sheep red blood cells and promotion of cell adhesion to a collagen substrate.


Progesterone-induced changes in sleep in male subjects.
Progesterone administration induces a reduction of the vigilance state in humans during wakefulness. It has been been suggested that this effect is mediated via neuroactive metabolites that interact with the gamma-aminobutyric, acidA (GABAA) receptor complex. To investigate the effects of progesterone administration on the sleep electroencephalogram (EEG) in humans we made polysomnographic recordings, including sleep stage-specific spectral analysis, and concomitantly measured plasma concentrations of progesterone and its GABA-active metabolites 3 alpha-hydroxy-5 alpha-dihydroprogesterone (allopregnanolone) and 3 alpha-hydroxy-5 beta-dihydroprogesterone (pregnanolone) in nine healthy male subjects in a double-blind placebo-controlled crossover study. Progesterone administration at 9:30 PM induced a significant increase in the amount of non-rapid eye movement (REM) sleep. The EEG spectral power during non-REM sleep showed a significant decrease in the slow wave frequency range (0.4-4.3 Hz), whereas the spectral power in the higher frequency range (> 15 Hz) tended to be elevated. Some of the observed changes in sleep architecture and sleep-EEG power spectra are similar to those induced by agonistic modulators of the GABAA receptor complex and appear to be mediated in part via the conversion of progesterone into its GABA-active metabolites.


Significant clinical response of progressive recurrent ovarian clear cell carcinoma to glypican-3-derived peptide vaccine therapy: two case reports.
Carcinoembryonic antigen glypican-3 (GPC3) is expressed by>40% of ovarian clear cell carcinoma (CCC) and is a promising immunotherapeutic target. We previously reported the safety of and immunological and clinical responses to a GPC3-derived peptide vaccine in a phase I clinical trial of patients with advanced hepatocellular carcinoma (HCC). Although the efficacy of the GPC3-derived peptide vaccine against HCC patients was evaluated, other GPC3-positive cancer patients have not yet been investigated. Therefore, we conducted a phase II trial to evaluate the clinical outcome of ovarian CCC patients treated with a GPC3-derived peptide vaccine. The GPC3 peptide was administered at a dose of 3 mg per body. Patients received an intradermal injection of the GPC3 peptide emulsified with incomplete Freund's adjuvant. Vaccinations were performed biweekly from the first until the 6th injection and were then repeated at 6-week intervals after the 7th injection. Treatment continued until disease progression. We herein present two patients with chemotherapy-refractory ovarian CCC who achieved a significant clinical response in an ongoing trial of a GPC3 peptide vaccine. Case 1, a 42-year-old patient with advanced recurrent ovarian CCC with liver and retroperitoneal lymph node metastases, received the HLA-A24-restricted GPC3 peptide vaccine. Contrast-enhanced CT at week 10 revealed a partial response (PR) using RECIST criteria. Case 2 was a 67-year-old female with multiple lymph node metastases. She was injected with the HLA-A2-restricted GPC3 peptide vaccine. According to RECIST, PR was achieved at week 37. The stabilization of their diseases over one year provided us with the first clinical evidence to demonstrate that GPC3 peptide-based immunotherapy may significantly prolong the overall survival of patients with refractory ovarian CCC.


Is the Suppressor-mutator element controlled by a basic developmental regulatory mechanism?
We report the results of genetic studies on derivatives of two different alleles of the maize a locus with an insertion of the Suppressor-mutator (Spm) transposable element in which the element is inactive, but can be reactivated readily. We present evidence that the mechanism that determines whether the element is in an active or inactive phase has two genetically distinguishable components. One determines whether or not the element is genetically active (the phase setting) and the other determines the stability of the setting in development, its heritability, and its phase in the next generation (the phase program). We show that the element's phase can be reset in a reproducible pattern during plant development. We also show that the Spm element can be reprogrammed to undergo a subsequent phase change without a concomitant phase change. The capacity to reset and reprogram the Spm element is differentially expressed within the plant in a pattern that is correlated with the developmental fate of apical and lateral meristems, suggesting the involvement of a basic developmental determination mechanism.


TRPA1-expressing lamina propria mesenchymal cells regulate colonic motility.
The physiological process of defecation is directly controlled by colorectal motility. The transient receptor potential ankyrin 1 (TRPA1) channel is expressed in small intestine enterochromaffin cells and is involved in gastrointestinal motility via serotonin release. In the colorectum, however, enterochromaffin cell localization is largely distinct from that in the small intestine. Here, we investigated the role of lower gastrointestinal tract TRPA1 in modulating colorectal motility. We found that in colonic tissue, TRPA1 is predominantly expressed in mesenchymal cells of the lamina propria, which are clearly distinct from those in the small intestine. These cells coexpressed COX1 and microsomal prostaglandin E synthase-1. Intracolonic administration of TRPA1 agonists induced colonic contraction, which was suppressed by a prostaglandin E2 (PGE2) receptor 1 antagonist. TRPA1 activation induced calcium influx and PGE2 release from cultured human fibroblastic cells. In dextran sulfate sodium-treated animals, both TRPA1 and its endogenous agonist were dramatically increased in the colonic lamina propria, accompanied by abnormal colorectal contractions. Abnormal colorectal contractions were significantly prevented by pharmacological and genetic inhibition of TRPA1. In conclusion, in the lower gastrointestinal tract, mesenchymal TRPA1 activation results in PGE2 release and consequently promotes colorectal contraction, representing what we believe is a novel physiological and inflammatory bowel disease-associated mechanism of gastrointestinal motility.


Human telomerase catalyzes nucleolytic primer cleavage.
Telomerase is a reverse transcriptase that uses an integral RNA molecule to add de novo G-rich repeats onto telomeric DNA, or onto nontelomeric DNA generated during chromosome fragmentation and breakage events. A telomerase-mediated DNA substrate cleavage activity has been reported in ciliates and yeasts. Nucleolytic cleavage may serve a proofreading function, enhance processivity or ensure that nontemplate telomerase RNA sequences are not copied into DNA. We identified and characterized a human telomerase-mediated nucleolytic cleavage activity using enzyme reconstituted in a rabbit reticulocyte lysate in vitro transcription/translation system and native enzyme extracted from cells. We found that telomerase catalyzed the removal of nucleotides from DNA substrates including those that can form a mismatch with the RNA template or that contain nontelomeric sequences located 3' to a telomeric sequence. Unlike Tetrahymena telomerase, human telomerase catalyzed the removal of more than one nucleotide (up to 13) from telomeric primers. DNA substrates predicted to align at the 3'-end of the RNA template were not cleaved, consistent with cleavage being dictated by the template 5'-end. We also found some differences in the nuclease activity between RRL-reconstituted human telomerase and native enzyme.


Surgical Treatment of Congenital and Obligatory Dislocation of the Patella in Children.
Congenital (fixed) dislocations and obligatory (habitual) patellar dislocations represent a complex clinical and surgical challenge. Numerous treatment options, offering different perspectives, and surgical solutions are reported in the literature.We implemented the surgical technique principles, originally described by Stanisavljevic, for congenital (fixed) and obligatory (habitual) patellar dislocations, with slight modifications. We retrospectively evaluated the results of group of 12 patients (15 knees), operated between the years 2002 and 2013. The procedure includes extensive subperiosteal quadriceps realignment and soft tissue medial plication, followed by distal realignment through patellar tendon splitting. The outcome measures were patellar stability, knee range of motion, Pedi-IKDC knee function score, and the PODCI global function score. The study group included 12 patients, 9 females and 3 males (15 knees) whose mean age was 5 years 2 months at diagnosis and 7 years 5 months at surgery. Nine patients had an underlying diagnosed genetic background [Down syndrome (n=6), Larsen syndrome (n=2), nail-patella syndrome (n=1)]. The mean follow-up was 46.2 months. Eleven patients, gained stable patella with no recurrence of dislocation. Postoperative knee active extension was improved significantly (P<0.0001) for all patients. The average postoperative Pedi-IKDC and PODCI scores were significantly higher (P<0.001) among the idiopathic group. These results suggest that the Stanisavljevic procedure principles with modifications, provides an efficient method for treatment of fixed and obligatory patellar dislocation. In our opinion, young age at surgery played an important role in the surgical outcome. Level IV-therapeutic.


Coarctation of the Aorta Stent Trial (COAST): study design and rationale.
Coarctation of the aorta (CoA) accounts for 4% to 5% of congenital cardiac abnormalities. Stent therapy has become an accepted alternative to surgery for older children and adults, although there are no balloon-expandable stents approved by the Food and Drug Administration for use in the aorta. The Cheatham-Platinum (CP) stent was designed for CoA therapy and is widely used outside the United States. We have designed the first prospective trial of stent therapy for CoA to serve as the pivotal trial for Food and Drug Administration approval of the CP stent. The COAST study is a prospective, multicenter, single-arm clinical study. The population includes patients with native or recurrent CoA. Four primary outcome variables were defined. For each variable, the stent will be compared to performance guidelines derived from surgical experience. The first efficacy outcome is reduction in arm-leg systolic blood pressure gradient, and the second is reduction in hospital length of stay. Safety outcomes include the following: the occurrence of any serious or somewhat serious adverse event attributed to the stent or implantation procedure and the occurrence of postprocedure paradoxical hypertension. A total of 105 patients treated with the CP stent will be enrolled. To ascertain the effectiveness and safety of an interventional device, randomized controlled trials have been offered as the criterion standard. However, these trials are not well suited to study rare conditions such as CoA, especially once the therapy in question has received acceptance within the medical community. New clinical trial and statistical approaches are needed to evaluate such therapies. The COAST study is an example of this kind of innovative trial design.


Dose finding double-blind clinical trial with praziquantel in schistosomiasis japonica patients.
A double-blind clinical trial with praziquantel was carried out. A total of 400 cases was treated with four different dosages, namely, 60, 50, 40 and 30 mg/kg body weight of praziquantel. The drug was given in one day divided into two doses. Identical placebo tablets were used to make up a total of 60 mg/kg. Tolerance was good in all with the exception of one case suffering from asthmatic attack with papule rashes over large area of the body surface. 394 patients were able to be followed up parasitologically six months post-treatment. 79.8%, 71.7%, 78.8% and 70.1% of the patients were negative in the groups with the total dose of 60, 50, 40 and 30 mg/kg respectively. The cure rates as well as the side effects were similar for the four groups. The efficacy was lower than that reported by other authors and the possibility of reinfection was incriminated. In villages with few snails the negative hatching rates in aforementioned four groups were 89.1%, 91.1%, 88.9% and 81.8%, while in villages with abundant snails the rates were 68.2%, 46.5%, 66.7% and 54.8%. The difference between the two areas was statistically significant. Higher efficacy was observed in adults with an average cure rate of 80.0% than in children under 15 years of age, the average cure rate being 57.1%.(ABSTRACT TRUNCATED AT 250 WORDS)