Sphingosine-1-phosphate improves endothelialization with reduction of thrombosis in recellularized human umbilical vein graft by inhibiting syndecan-1 shedding in vitro.
Sphingosine-1-phosphate (S1P) has been known to promote endothelial cell (EC) proliferation and protect Syndecan-1 (SDC1) from shedding, thereby maintaining this antithrombotic signal. In the present study, we investigated the effect of S1P in the construction of a functional tissue-engineered blood vessel by using human endothelial cells and decellularized human umbilical vein (DHUV) scaffolds. Both human umbilical vein endothelial cells (HUVEC) and human cord blood derived endothelial progenitor cells (EPC) were seeded onto the scaffold with or without the S1P treatment. The efficacy of re-cellularization was determined by using the fluorescent marker CellTracker CMFDA and anti-CD31 immunostaining. The antithrombotic effect of S1P was examined by the anti-aggregation tests measuring platelet adherence and clotting time. Finally, we altered the expression of SDC1, a major glycocalyx protein on the endothelial cell surface, using MMP-7 digestion to explore its role using platelet adhesion tests in vitro. The result showed that S1P enhanced the attachment of HUVEC and EPC. Based on the anti-aggregation tests, S1P-treated HUVEC recellularized vessels when grafted showed reduced thrombus formation compared to controls. Our results also identified reduced SDC1 shedding from HUVEC responsible for inhibition of platelet adherence. However, no significant antithrombogenic effect of S1P was observed on EPC. In conclusion, S1P is an effective agent capable of decreasing thrombotic risk in engineered blood vessel grafts. Sphingosine-1phosphate (S1P) is a low molecular-weight phospholipid mediator that regulates diverse biological activities of endothelial cell, including survival, proliferation, cell barrier integrity, and also influences the development of the vascular system. Based on these characters, we the first time to use it as an additive during the process of a small caliber blood vessel construction by decellularized human umbilical vein and endothelial cell/endothelial progenitor. We further explored the function and mechanism of S1P in promoting revascularization and protection against thrombosis in this tissue engineered vascular grafts. The results showed that S1P could not only accelerate the generation but also reduce thrombus formation of small caliber blood vessel.


How to ask for and act on feedback: practical tips for medical students.
This article was migrated. The article was marked as recommended. Receiving feedback is an essential component of learning and can be invaluable to help you improve your knowledge and skills as you progress towards qualification as a doctor. Whilst formal feedback from assessments is commonplace, it can be daunting to approach others to provide you with feedback. Furthermore, it can be difficult to know how to act on the feedback once you have received it. In this article we look at how best for medical students to ask for feedback and act on the advice which is given to them with a unique point-of-view provided from a current clinical teaching fellow.


[The mechanisms of the lymph-stimulating action of liposomes].
The mechanisms of lymph-stimulating action of different liposomes (lecithin-cholesterin, lecithin) were studied in the experiments on dogs. It has been shown an absence of interaction between changes in the lymph outflow rate and system circulation parameters when injecting any liposomes. Moreover, water and protein filtration from the vascular bed into the interstitial space was lowered due to liposomal injection. But oxygen tension in the muscular tissues remained on the level closed to the normal one. In contrast to the intravenous injection, the subcutaneous administration of liposomes has induced the stimulation of lymph outflow, which was more expressed on both volume (nearly by 40%) and duration (nearly by 50%). The experiments with limited and crossed blood flow have demonstrated a regional character of liposomal lymph-stimulating. The mechanisms of such liposomal action may be related to both direct action of liposomes on the lymphatic smooth muscle contractility in the lymphatic vessels and the increased lymph production in the site of liposomal injection due to free radical oxidation processes being limited in that area. The micellar form of phospholipids (essential) has demonstrated no lymph-stimulating action.


Influence of CYP2C8 polymorphisms on imatinib steady-state trough level in chronic myeloid leukemia and gastrointestinal stromal tumor patients.
Imatinib trough levels have been associated with its clinical effects. During chronic use of imatinib, CYP2C8 becomes an important metabolizing enzyme because of cytochrome P450 3A4 (CYP3A4) autoinhibition. Single nucleotide polymorphisms (SNPs) in CYP2C8 may affect imatinib trough levels. This study investigates the effect of common CYP2C8 polymorphisms [*1B (rs7909236), *1C (rs17110453), *3 (rs11572080 and rs10509681), and *4 (rs1058930)] on steady-state trough levels imatinib during chronic imatinib use in 43 patients with chronic myeloid leukemia or gastrointestinal stromal tumors. Standardized imatinib trough levels did not show a significant difference between wild-type and variant groups for any of the tested SNPs, but an association with age was found, with older patients having higher trough levels. This suggests that common CYP2C8 SNPs have no effect on the pharmacokinetics of imatinib.


Characterization of recycled mushroom compost leachate by chemical analysis and thermogravimetry-mass spectrometry.
Recycled compost leachate (RCL or euphemistically named "goody water") can be a potent source of foul odor on mushroom substrate production sites and contributes to composting smells. A complex mixture of sulfur compounds, fatty acids, and nitrogen containing compounds is responsible for odor production. Fifty samples, collected from 14 compost production sites in Ireland and the U.K. over a 2 year period, were analyzed for chemical properties and by thermogravimetry-mass spectrometry (TG-MS) for compositional differences. Results indicated that aerated samples had lower values of electrical conductivity, redox potential, and dry matter content than nonaerated samples and that the higher thermal stability of aerated samples measured by TGA could be attributed to greater mineralization of the substrate due to aerobic processes. The lower temperatures noted for peak evolution of methane, water, and carbon dioxide from TG-MS analysis suggested that a more energetic process had occurred in aerated RCL storage facilities, producing greater decomposition of macromolecules that volatilized at lower temperatures. Chemical composition, thermal stability of the freeze-dried leachate, pyrolysis profiles, and relative amounts of pyrolysis products were all markers of as to how effective control measures could influence RCL quality.


Design Optimization and Structural Performance Evaluation of Plate Girder Bridge Constructed Using a Turn-Over Process.
A recent trend in bridge construction has been the optimization of the cost-to-performance ratio. The most effective way to optimize the cost-to-performance ratio is to maximize the efficiency of the superstructure. Currently, many bridge engineers and designers favor two- or three- girder plate superstructures, due to their cost advantages. However, research on the performance enhancements of the I-type girder in two- or three- girder plate bridges is lacking. One of the most important performance improvement technologies for the I-type girder is the "preflex" method. In the preflex method, the specimen is inverted during the construction process to apply prestressed cambering to the specimen by using self-weight. However, a problem with the preflex construction method is difficulty with inverting the girder/plate system during the concrete curing process. Therefore, a new inverting system called Turn-Over (TO) wheel was proposed. Using TO wheels, wider variations to the I-type girder design can be achieved. Using this TO construction method, various cross sectional designs of girder plate systems can be considered due to its easiness in inverting the girder/plate system. In this study, the location of concrete confinement sections between the steel I-beams and concrete plates was varied in an I-girder cross-sectional design. Design parameters included effective height, flange thickness, flange width, confining concrete section width, etc. From this study, the optimum cross-sectional design of the I-girder/concrete plate system was achieved. Then, a single 20 m TO girder/plate system and two 20 m TO girder bridges were constructed and tested to evaluate their performance. From the test, failure behavior, load carrying capacity, crack pattern, etc., are obtained. The results are discussed in detail in this paper.


Fibroblast Growth Factor 19 modulates intestinal microbiota and inflammation in presence of Farnesoid X Receptor.
Disruption of bile acid (BA) homeostasis plays a key role in intestinal inflammation. The gut-liver axis is the main site for the regulation of BA synthesis and BA pool size via the combined action of the nuclear Farnesoid X Receptor (FXR) and the enterokine Fibroblast Growth Factor 19 (FGF19). Increasing evidence have linked derangement of BA metabolism with dysbiosis and mucosal inflammation. Thus, here we aimed to investigate the potential action of an FGF19 analogue on intestinal microbiota and inflammation. A novel engineered non-tumorigenic variant of the FGF19 protein, M52-WO 2016/0168219 was generated. WT and FXRnull mice were injected with AAV-FGF19-M52 or the control AAV-GFP and subjected to Sodium Dextran Sulphate-induced colitis. FGF19-M52 reduced BA synthesis and pool size, modulated its composition and protected mice from intestinal inflammation. These events were coupled with preservation of the intestinal epithelial barrier integrity, inhibition of inflammatory immune response and modulation of microbiota composition. Interestingly, FGF19-M52-driven systemic and local anti-inflammatory activity was completely abolished in Farnesoid X Receptor (FXR)null mice, thus underscoring the need of FXR to guarantee enterocytes' fitness and complement FGF19 anti-inflammatory activity. To provide a translational perspective, we also show that circulating FGF19 levels are reduced in patients with Crohn's disease. Reactivation of the FXR-FGF19 axis in a murine model of intestinal inflammation could bona fide provide positive changes in BA metabolism with consequent reduction of intestinal inflammation and modulation of microbiota. These results point to the therapeutic potential of FGF19 in the treatment of intestinal inflammation with concomitant derangement of BA homeostasis. A. Moschetta is funded by MIUR-PRIN 2017 <- 2017J3E2W2; Italian Association for Cancer Research (AIRC, IG 23239); Interreg V-A Greece-Italy 2014-2020-SILVER WELLBEING, MIS5003627; HDHL-INTIMIC EuJPI-FATMAL; MIUR PON "R&I" 2014-2020-ARS01_01220. No money has been paid by NGM Biopharmaceuticals or any other agency to write this article.


Evaluation of a capacity building clinical educational model for oral health clinicians treating very young children.
There are significant levels of dental caries in Australian school-aged children, with children aged five years having a mean dmft of 1.3. It has also been identified that, in general, oral health clinicians lack confidence to treat very young children and this study aimed to increase capacity of public sector oral health clinicians to treat preschool children. An educational program was developed, implemented and evaluated for its capability to increase the confidence and knowledge of oral health clinicians and dental assistants in providing oral care for children aged 12 months to 5 years. In 2011 and 2012, the course was delivered to 36 clinicians (22 dentists, 12 dental therapists, and two oral health therapists) and showed increases in their confidence and knowledge for participants when providing dental procedures to preschool children. The educational program that was developed and implemented has met its objective of increasing the knowledge and confidence of practicing oral health clinicians and dental assistants in the management of preschool children. Strategies to further enhance the outcomes of this educational program have been proposed.


Beta thromboglobulin and platelet factor 4 in bronchoalveolar lavage fluid of patients with systemic sclerosis.
To evaluate concentrations of the platelet activation markers beta thromboglobulin (BTG) and platelet factor 4 (PF4) in bronchoalveolar lavage fluid (BALF) from patients with systemic sclerosis with and without scleroderma interstitial lung disease (SLD). BTG and PF-4 were measured by enzyme immunoassay in BALF from 37 patients with systemic sclerosis. Controls were 10 healthy subjects. BALF was collected during routine bronchoscopy from the right middle lobe. SLD was diagnosed by high resolution computed tomography of the lungs. BTG was detected in 11 of the patients with systemic sclerosis (29.7%) and PF4 was found in eight (21.6%). Mean (SD) concentrations of BTG and PF4 in BALF from patients with detectable levels of these platelet activation markers were 106.9 (69.8) and 35.2 (17.4) IU/ml, respectively. The BTG:PF4 ratio was more than 2:1, indicating in vivo release. Both markers were found exclusively in patients with SLD. SLD patients with detectable platelet activation markers had a significantly shorter disease duration than those with undetectable BTG/PF4. The study provides evidence that activation of blood platelets takes place within the lungs of patients with SLD and may contribute to the development of lung fibrosis.


Pseudomonas aeruginosa inactivation mechanism is affected by capsular extracellular polymeric substances reactivity with chlorine and monochloramine.
The reactivity of capsular extracellular polymeric substances (EPS) to chlorine and monochloramine was assessed and compared in this study. The impact of capsular EPS on Gram-negative bacteria Pseudomonas aeruginosa inactivation mechanisms was investigated both qualitatively and quantitatively using a combination of batch experiments, viability tests with LIVE/DEAD staining, and Fourier transform infrared spectroscopy (FTIR). Both wild-type and isogenic mutant strains with different alginate EPS production capabilities were used to evaluate their susceptibility to chlorine and monochloramine. The mucA22 mutant strain, which overproduces the EPS composed largely of acidic polysaccharide alginate, exhibited high resistance and prolonged inactivation time to both chlorine and monochloramine relative to PAO1 (wild-type) and algT(U) mutant strains (alginate EPS deficient). Multiple analyses were combined to better understand the mechanistic role of EPS against chlorine-based disinfectants. The extracted EPS exhibited high reactivity with chlorine and very low reactivity with monochloramine, suggesting different mechanism of protection against disinfectants. Moreover, capsular EPS on cell membrane appeared to reduce membrane permeabilization by disinfectants as suggested by deformation of key functional groups in EPS and cell membrane (the C-O-C stretching of carbohydrate and the C=O stretching of ester group). The combined results supported that capsular EPS, acting either as a disinfectant consumer (for chlorine inactivation) or limiting access to reactive sites on cell membrane (for monochloramine inactivation), provide a protective role for bacterial cells against regulatory residual disinfectants by reducing membrane permeabilization.


Salmonid alphavirus glycoprotein E2 requires low temperature and E1 for virion formation and induction of protective immunity.
Salmonid alphavirus (SAV; also known as Salmon pancreas disease virus; family Togaviridae) causes pancreas disease and sleeping disease in Atlantic salmon and rainbow trout, respectively, and poses a major burden to the aquaculture industry. SAV infection in vivo is temperature-restricted and progeny virus is only produced at low temperatures (10-15 °C). Using engineered SAV replicons we show that viral RNA replication is not temperature-restricted suggesting that the viral structural proteins determine low-temperature dependency. The processing/trafficking of SAV glycoproteins E1 and E2 as a function of temperature was investigated via baculovirus vectors in Sf9 insect cells and by transfection of CHSE-214 fish cells with DNA constructs expressing E1 and E2. We identified SAV E2 as the temperature determinant by demonstrating that membrane trafficking and surface expression of E2 occurs only at low temperature and only in the presence of E1. Finally, a vaccination-challenge model in Atlantic salmon demonstrates the biological significance of our findings and shows that SAV replicon DNA vaccines encoding E2 elicit protective immunity only when E1 is co-expressed. This is the first study that identifies E2 as the critical determinant of SAV low-temperature dependent virion formation and defines the prerequisites for induction of a potent immune response in Atlantic salmon by DNA vaccination.


Vascular cell adhesion molecule-1 (VCAM-1) plays a central role in the pathogenesis of severe forms of vasculitis due to hepatitis C-associated mixed cryoglobulinemia.
To better characterize the molecules involved in leukocyte tissue infiltration during hepatitis C-mixed cryoglobulinemia (HCV-MC)-associated vasculitis. The involvement of ELAM, ICAM-1 and VCAM-1 was evaluated in 36 patients with HCV-MC vasculitis using three different approaches: concentrations of soluble forms by specific ELISA, tissue expression by immunohistochemistry on patients nerve biopsies, endothelial expression by FACS analysis, on cells activated in vitro by cryoprecipitates purified from HCV-MC patients. Concentrations of sVCAM-1 were significantly elevated in the serum of HCV-MC patients compared to HCV patients without MC, the highest concentrations being found in severe vasculitis. VCAM-1 expression was detected on blood vessels from nerve biopsies performed in patients with severe vasculitis. When added to endothelial cells in vitro, HCV-MC patients cryoprecipitate induced VCAM-1 but also ELAM and ICAM-1 expression possibly through a mechanism due to the C1q complement fraction interaction with endothelial cells, since C1q was consistently present in the cryoprecipitates. VCAM-1 is mainly involved in the pathogenesis of HCV-MC-associated severe vasculitis and may be a potential interesting therapeutic target.


Site-Dependent Relationships Between Fungal Community Composition, Plant Genotypic Diversity and Environmental Drivers in a Salix Biomass System.
Soil fungi are strongly affected by plant species or genotypes since plants modify their surrounding environment, but the effects of plant genotype diversity on fungal diversity and function have not been extensively studied. The interactive responses of fungal community composition to plant genotypic diversity and environmental drivers were investigated in Salix biomass systems, posing questions about: (1) How fungal diversity varies as a function of plant genotype diversity; (2) If plant genotype identity is a strong driver of fungal community composition also in plant mixtures; (3) How the fungal communities change through time (seasonally and interannually)?; and (4) Will the proportion of ECM fungi increase over the rotation? Soil samples were collected over 4 years, starting preplanting from two Salix field trials, including four genotypes with contrasting phenology and functional traits, and genotypes were grown in all possible combinations (four genotypes in Uppsala, Sweden, two in Rostock, Germany). Fungal communities were identified, using Pacific Biosciences sequencing of fungal ITS2 amplicons. We found some site-dependent relationships between fungal community composition and genotype or diversity level, and site accounted for the largest part of the variation in fungal community composition. Rostock had a more homogenous community structure, with significant effects of genotype, diversity level, and the presence of one genotype ("Loden") on fungal community composition. Soil properties and plant and litter traits contributed to explaining the variation in fungal species composition. The within-season variation in composition was of a similar magnitude to the year-to-year variation. The proportion of ECM fungi increased over time irrespective of plant genotype diversity, and, in Uppsala, the 4-mixture showed a weaker response than other combinations. Species richness was generally higher in Uppsala compared with that in Rostock and increased over time, but did not increase with plant genotype diversity. This significant site-specificity underlines the need for consideration of diverse sites to draw general conclusions of temporal variations and functioning of fungal communities. A significant increase in ECM colonization of soil under the pioneer tree Salix on agricultural soils was evident and points to changed litter decomposition and soil carbon dynamics during Salix growth.


Differences in brain processing of proprioception related to postural control in patients with recurrent non-specific low back pain and healthy controls.
Patients with non-specific low back pain (NSLBP) show an impaired postural control during standing and a slower performance of sit-to-stand-to-sit (STSTS) movements. Research suggests that these impairments could be due to an altered use of ankle compared to back proprioception. However, the neural correlates of these postural control impairments in NSLBP remain unclear. Therefore, we investigated brain activity during ankle and back proprioceptive processing by applying local muscle vibration during functional magnetic resonance imaging in 20 patients with NSLBP and 20 controls. Correlations between brain activity during proprioceptive processing and (Airaksinen et al., 2006) proprioceptive use during postural control, evaluated by using muscle vibration tasks during standing, and (Altmann et al., 2007) STSTS performance were examined across and between groups. Moreover, fear of movement was assessed. Results revealed that the NSLBP group performed worse on the STSTS task, and reported more fear compared to healthy controls. Unexpectedly, no group differences in proprioceptive use during postural control were found. However, the relationship between brain activity during proprioceptive processing and behavioral indices of proprioceptive use differed significantly between NSLBP and healthy control groups. Activity in the right amygdala during ankle proprioceptive processing correlated with an impaired proprioceptive use in the patients with NSLBP, but not in healthy controls. Moreover, while activity in the left superior parietal lobule, a sensory processing region, during back proprioceptive processing correlated with a better use of proprioception in the NSLBP group, it was associated with a less optimal use of proprioception in the control group. These findings suggest that functional brain changes during proprioceptive processing in patients with NSLBP may contribute to their postural control impairments.


Laparoscopic versus open appendectomy: a prospective randomized double-blind study.
The value of laparoscopy in appendicitis is not established. Studies suffer from multiple limitations. Our aim is to compare the safety and benefits of laparoscopic versus open appendectomy in a prospective randomized double blind study. Two hundred forty-seven patients were analyzed following either laparoscopic or open appendectomy. A standardized wound dressing was applied blinding both patients and independent data collectors. Surgical technique was standardized among 4 surgeons. The main outcome measures were postoperative complications. Secondary outcome measures included evaluation of pain and activity scores at base line preoperatively and on every postoperative day, as well as resumption of diet and length of stay. Activity scores and quality of life were assessed on short-term follow-up. There was no mortality. The overall complication rate was similar in both groups (18.5% versus 17% in the laparoscopic and open groups respectively), but some early complications in the laparoscopic group required a reoperation. Operating time was significantly longer in the laparoscopic group (80 minutes versus 60 minutes; P = 0.000) while there was no difference in the pain scores and medications, resumption of diet, length of stay, or activity scores. At 2 weeks, there was no difference in the activity or pain scores, but physical health and general scores on the short-form 36 (SF36) quality of life assessment forms were significantly better in the laparoscopic group. Appendectomy for acute or complicated (perforated and gangrenous) appendicitis had similar complication rates, regardless of the technique (P = 0.181). Unlike other minimally invasive procedures, laparoscopic appendectomy did not offer a significant advantage over open appendectomy in all studied parameters except quality of life scores at 2 weeks. It also took longer to perform. The choice of the procedure should be based on surgeon or patient preference.


Transplant-Associated Thrombotic Microangiopathy in Pediatric Hematopoietic Cell Transplant Recipients: A Practical Approach to Diagnosis and Management.
Transplant-associated thrombotic microangiopathy (TA-TMA) is an endothelial damage syndrome that is increasingly identified as a complication of both autologous and allogeneic hematopoietic cell transplantation (HCT) in children. If not promptly diagnosed and treated, TA-TMA can lead to significant morbidity (e.g., permanent renal injury) or mortality. However, as the recognition of the early stages of TA-TMA may be difficult, we propose a TA-TMA "triad" of hypertension, thrombocytopenia (or platelet transfusion refractoriness), and elevated lactate dehydrogenase (LDH). While not diagnostic, this triad should prompt further evaluation for TA-TMA. There is increased understanding of the risk factors for the development of TA-TMA, including those which are inherent (e.g., race, genetics), transplant approach-related (e.g., second HCT, use of HLA-mismatched donors), and related to post-transplant events (e.g., receipt of calcineurin inhibitors, development of graft-vs. -host-disease, or certain infections). This understanding should lead to enhanced screening for TA-TMA signs and symptoms in high-risk patients. The pathophysiology of TA-TMA is complex, resulting from a cycle of activation of endothelial cells to produce a pro-coagulant state, along with activation of antigen-presenting cells and lymphocytes, as well as activation of the complement cascade and microthrombi formation. This has led to the formulation of a "Three-Hit Hypothesis" in which patients with either an underlying predisposition to complement activation or pre-existing endothelial injury (Hit 1) undergo a toxic conditioning regimen causing endothelial injury (Hit 2), and then additional insults are triggered by medications, alloreactivity, infections, and/or antibodies (Hit 3). Understanding this cycle of injury permits the development of a specific TA-TMA treatment algorithm designed to treat both the triggers and the drivers of the endothelial injury. Finally, several intriguing approaches to TA-TMA prophylaxis have been identified. Future work on the development of a single diagnostic test with high specificity and sensitivity, and the development of a robust risk-scoring system, will further improve the management of this serious post-transplant complication.


Techniques for framing questions in conducting family meetings in palliative care.
Family therapy has developed several approaches to framing questions within family meetings, but few of these techniques have been adapted for palliative care. We focus on the application of questioning techniques from systemic family therapy to palliative care. More specifically, we describe and give examples of the model of asking questions developed by Karl Tomm (1988) through its application in Family Focused Grief Therapy (FFGT), a preventive intervention delivered to high-risk families during palliative care and bereavement. First, the type of questions used across the course of therapy is explored based on the interventive questioning model. Then, a case example is provided to demonstrate the use and adaptation of this model in a palliative care setting. At the beginning of therapy, the most frequent questions were linear and circular, moving around the family to build up a picture of events from everyone's perspective. As for the frequency of reflexive and strategic questions, these increased as the therapy progressed, bringing the family to new perspectives. The case example fleshes out the importance of each type of question, all of which have a proper place in the course of therapy. These illustrations highlight the value of having a model of questioning styles to guide the clinician when exploring palliative care issues, such as care provision, coping and grief, intimacy, and discussing death. This framework could be useful in guiding supervisors, trainees, and clinicians seeking to build skills and optimize their interventions in a palliative care setting.


Common mental disorders in hematopoietic stem cell transplant patients: a scoping review.
to map common recurrent mental disorders in patients undergoing hematopoietic stem cell transplantation. this is a scoping review carried out in January 2022 in electronic databases and repositories of dissertations and thesis. Studies that answered the research question, met the objective of the study and were available in full electronically, in any language, were included. the sample consisted of 28 studies, 14 of which were published in the United States of America. The common mental disorders found were depressive, anxiety, post-traumatic stress and mood disorders. Twenty symptoms were mentioned, among the most prevalent are fatigue and sleep disorders/insomnia. the difficulty and importance of carrying out the differential diagnosis of these disorders were highlighted, since their symptoms can be confused with other health problems and have a strong potential to interfere with patients' evolution.


Automatic image segmentation by dynamic region merging.
This paper addresses the automatic image segmentation problem in a region merging style. With an initially oversegmented image, in which many regions (or superpixels) with homogeneous color are detected, an image segmentation is performed by iteratively merging the regions according to a statistical test. There are two essential issues in a region-merging algorithm: order of merging and the stopping criterion. In the proposed algorithm, these two issues are solved by a novel predicate, which is defined by the sequential probability ratio test and the minimal cost criterion. Starting from an oversegmented image, neighboring regions are progressively merged if there is an evidence for merging according to this predicate. We show that the merging order follows the principle of dynamic programming. This formulates the image segmentation as an inference problem, where the final segmentation is established based on the observed image. We also prove that the produced segmentation satisfies certain global properties. In addition, a faster algorithm is developed to accelerate the region-merging process, which maintains a nearest neighbor graph in each iteration. Experiments on real natural images are conducted to demonstrate the performance of the proposed dynamic region-merging algorithm.


Platelet-activating factor-induced chemokine gene expression requires NF-kappaB activation and Ca2+/calcineurin signaling pathways. Inhibition by receptor phosphorylation and beta-arrestin recruitment.
Previously, we reported that platelet-activating factor (PAF) stimulates higher G protein activation and a more robust Ca2+ mobilization in RBL-2H3 cells expressing carboxyl terminus deletion, phosphorylation-deficient mutant of PAF receptor (mPAFR) when compared with the wild-type receptor (PAFR). However, PAF did not provide sufficient signal for CC chemokine receptor ligand 2 (CCL2) production in cells expressing mPAFR. Based on these findings, we hypothesized that receptor phosphorylation provides a G protein-independent signal that synergizes with Ca2+ mobilization to induce CCL2 production. Here, we show that a mutant of PAFR (D289A), which does not couple to G proteins, was resistant to agonist-induced receptor phosphorylation. Unexpectedly, we found that when this mutant was coexpressed with mPAFR, it restored NF-kappaB activation and CCL2 production. PAF caused translocation of beta-arrestin from the cytoplasm to the membrane in cells expressing PAFR but not a phosphorylation-deficient mutant in which all Ser/Thr residues were replaced with Ala (DeltaST-PAFR). Interestingly, PAF induced significantly higher NF-kappaB and nuclear factor of activated T cells (NFAT)-luciferase activity as well as CCL2 production in cells expressing DeltaST-PAFR than those expressing PAFR. Furthermore, a Ca2+/calcineurin inhibitor completely inhibited PAF-induced NFAT activation and CCL2 production but not NF-kappaB activation. These findings suggest that the carboxyl terminus of PAFR provides a G protein-independent signal for NF-kappaB activation, which synergizes with G protein-mediated Ca2+/calcineurin activation to induce CCL2 production. However, receptor phosphorylation and beta-arrestin recruitment inhibit CCL2 production by blocking both NF-kappaB activation and Ca2+/calcineurin-dependent signaling pathways.


Otolaryngology workforce analysis.
The number of trained otolaryngologists available is insufficient to supply current and projected US health care needs. The goal of this study was to assess available databases and present accurate data on the current otolaryngology workforce, examine methods for prediction of future health care needs, and explore potential issues with forecasting methods and policy implementation based on these predictions. Retrospective analysis of research databases, public use files, and claims data. The total number of otolaryngologists and current practices in the United States was tabulated using the databases of the American Academy of Otolaryngology-Head and Neck Surgery, American Medical Association, American Board of Otolaryngology, American College of Surgeons, Association of American Medical Colleges, National Center for Health Statistics, and Department of Health and Human Services. Otolaryngologists were identified as surgeons and classified into surgical groups using a combination of AMA primary and secondary self-reported specialties and American Board of Medical Specialties certifications. Data gathered were cross-referenced to rule out duplications to assess total practicing otolaryngologists. Data analyzed included type of practice: 1) academic versus private and 2) general versus specialty; and demographics: 1) urban versus rural, 2) patient age, 3) reason for visit (referral, new, established, surgical follow-up), 4) reason for visit (diagnosis), and 5) payer type. Analysis from the above resources estimates the total number of otolaryngologists practicing in the United States in 2011 to be 12,609, with approximately 10,522 fully trained practicing physicians (9,232-10,654) and 2,087 in training (1,318 residents and 769 fellows/others). Based on 2011 data, workforce projections would place the fully trained and practicing otolaryngology workforce at 11,088 in 2015 and 12,084 in 2025 unless changes in training occur. The AAO-HNS Physicians Resource Committee performed an extensive analysis of collated data from multiple sources in 2014 and identified 10,800 practicing otolaryngologists and 2,087 in training. It is estimated that the current attrition rate is approximately 306 otolaryngologists per year. Percentage distribution of office visits by patient age was found to be 20% <15 years old, 7% 15 to 24 years old, 21% 25 to 44 years old, 32% 45 to 64 years old, 11% 65 to 74 years old, and 10% ≥75 years old. Reason for visit was 34% new, 29% chronic, 17% chronic with exacerbation, and 15% pre- or postsurgical follow-up. The top diagnoses consisted of otitis media, chronic sinusitis, and impacted cerumen. Payer mix consisted of 59% private insurance, 19% Medicare, and 12% Medicaid/Children's Health Insurance Program. Despite past findings and predictions of 8,000 to 8,500 otolaryngologists practicing in the United States, collated data from above resources places the total at 12,887, with 10,800 fully trained and practicing in 2014. This 30% to 50% underestimation of the otolaryngology workforce has an impact on future predictions and resource utilization analysis. Even when this correction is considered, the available trained otolaryngologists required to serve the otolaryngologic health care needs of the US population are still insufficient and understaffed. The impact of an aging population and the estimated 30 to 47 million newly insured citizens under the 2010 Patient Protection and Affordable Care Act are also unprecedented variables that must be considered. Further analysis of differences in physician productivity and geographic population density, and model formation of current otolaryngology workforce utilization, are needed to predict future public health needs. NA Laryngoscope, 126:5-11, 2016.


Physiological effects of high-flow oxygen in tracheostomized patients.
High-flow oxygen therapy via nasal cannula (HFOTNASAL) increases airway pressure, ameliorates oxygenation and reduces work of breathing. High-flow oxygen can be delivered through tracheostomy (HFOTTRACHEAL), but its physiological effects have not been systematically described. We conducted a cross-over study to elucidate the effects of increasing flow rates of HFOTTRACHEAL on gas exchange, respiratory rate and endotracheal pressure and to compare lower airway pressure produced by HFOTNASAL and HFOTTRACHEAL. METHODS: Twenty-six tracheostomized patients underwent standard oxygen therapy through a conventional heat and moisture exchanger, and then HFOTTRACHEAL through a heated humidifier, with gas flow set at 10, 30 and 50 L/min. Each step lasted 30 min; gas flow sequence during HFOTTRACHEAL was randomized. In five patients, measurements were repeated during HFOTTRACHEAL before tracheostomy decannulation and immediately after during HFOTNASAL. In each step, arterial blood gases, respiratory rate, and tracheal pressure were measured. During HFOTTRACHEAL, PaO2/FiO2 ratio and tracheal expiratory pressure slightly increased proportionally to gas flow. The mean [95% confidence interval] expiratory pressure raise induced by 10-L/min increase in flow was 0.2 [0.1-0.2] cmH2O (ρ = 0.77, p < 0.001). Compared to standard oxygen, HFOTTRACHEAL limited the negative inspiratory swing in tracheal pressure; at 50 L/min, but not with other settings, HFOTTRACHEAL increased mean tracheal expiratory pressure by (mean difference [95% CI]) 0.4 [0.3-0.6] cmH2O, peak tracheal expiratory pressure by 0.4 [0.2-0.6] cmH2O, improved PaO2/FiO2 ratio by 40 [8-71] mmHg, and reduced respiratory rate by 1.9 [0.3-3.6] breaths/min without PaCO2 changes. As compared to HFOTTRACHEAL, HFOTNASAL produced higher tracheal mean and peak expiratory pressure (at 50 L/min, mean difference [95% CI]: 3 [1-5] cmH2O and 4 [1-7] cmH2O, respectively). As compared to standard oxygen, 50 L/min of HFOTTRACHEAL are needed to improve oxygenation, reduce respiratory rate and provide small degree of positive airway expiratory pressure, which, however, is significantly lower than the one produced by HFOTNASAL.


A cross-sectional study of awareness and practices regarding animal bites in rural community, North India.
Repeated epidemiological studies to monitor trends of knowledge and practices are needed to guide strategies to control rabies. We conducted a study to assess the current knowledge, attitudes, and practices in relation to animal bites in the rural area of north India. House to house survey to collect data on animal bites was conducted among 300 households (assuming awareness regarding animal bites to be 25%, precision 95%, and power of 80%) from the rural area of Punjab, north India. A pretested semi-structured questionnaire comprising of items that explored sociodemographic details (age, educational qualification, occupation, socioeconomic status (assessed through Udai Pareek scale), and awareness regarding rabies, knowledge about first aid, attitude, and practices regarding anti-rabies vaccination (ARV) was used. Detailed questions were asked to those who owned pets. A total of 300 households were included in the analysis. Among all respondents, 30.4% (117) had an episode of animal bite in their family giving a bite incidence rate of 78/1000 population. Bites were more frequent in males (65.8%, n = 77). The commonest site of the bite was lower limb (65%) followed by upper limb (21.4%), and head and neck (5.1%). The participants said that bites by pet animals (47%) are more common than those by stray animals (35.9%), followed by wild animals (12.8%). Almost 91% of respondents told that they would prefer govt. hospital for the treatment. Class I bite was most common (88.9%) followed by class II (8.5%) and class III (1.7%). A lot of respondents (41.4%) did not know about the symptoms of rabies in humans. Only 17.5% knew the appropriate wound care. Inappropriate practices like applying chilly (48.8%), lime (13.1%), tying the limb above the wound (5.1%), and others were common. Only 15.5% washed their wound with soap and water. Most of those who were bitten received post-exposure prophylaxis (PEP) (80%). Most of the respondents (98.3%) had heard about ARV but didn't know about the site of injection. Almost everyone (99.35) said that no awareness camps/programs had been conducted in their villages/school/health center to date. There is a high incidence of animal bites in rural areas. Awareness regarding the need for rabies vaccine of animals and PEP after an animal bite is quite high and is practiced. However, there is a lack of awareness regarding the course of action to be followed when an animal does develop rabies. Traditional and inappropriate practices of wound management persist and need to be countered. Improving the availability of ARV and rabies immunoglobulin through the public health system may further augment the uptake of PEP and completion of treatment while at the same time reducing out of pocket expenditure and the overall economic cost of rabies. Solid waste management in rural areas along with oral ARV is likely to reduce the incidence of rabies in rural areas.


Motion perception correlates with volitional but not reflexive eye movements.
Visually-driven actions and perception are traditionally ascribed to the dorsal and ventral visual streams of the cortical processing hierarchy. However, motion perception and the control of tracking eye movements both depend on sensory motion analysis by neurons in the dorsal stream, suggesting that the same sensory circuits may underlie both action and perception. Previous studies have suggested that multiple sensory modules may be responsible for the perception of low- and high-level motion, or the detection versus identification of motion direction. However, it remains unclear whether the sensory processing systems that contribute to direction perception and the control of eye movements have the same neuronal constraints. To address this, we examined inter-individual variability across 36 observers, using two tasks that simultaneously assessed the precision of eye movements and direction perception: in the smooth pursuit task, observers volitionally tracked a small moving target and reported its direction; in the ocular following task, observers reflexively tracked a large moving stimulus and reported its direction. We determined perceptual-oculomotor correlations across observers, defined as the correlation between each observer's mean perceptual precision and mean oculomotor precision. Across observers, we found that: (i) mean perceptual precision was correlated between the two tasks; (ii) mean oculomotor precision was correlated between the tasks, and (iii) oculomotor and perceptual precision were correlated for volitional smooth pursuit, but not reflexive ocular following. Collectively, these results demonstrate that sensory circuits with common neuronal constraints subserve motion perception and volitional, but not reflexive eye movements.


Complication rate of the fascia closure technique in endovascular aneurysm repair.
To assess the rate of complications associated with the fascia closure technique for femoral access sites in which 18-F or 20-F sheaths were introduced during endovascular aneurysm repair (EVAR). A retrospective analysis was done of 50 consecutive patients (41 men; median age 75 years, range 62-85) who received Excluder stent-grafts in planned percutaneous EVAR procedures from May 2006 until December 2009. The fascia closure technique was routinely used for all femoral access sites in which large bore (18-F and 20-F) introducers were employed. One patient with extremely calcified and narrowed vessels was converted to primary cutdown bilaterally after percutaneous access failed. In the 49 remaining patients, 81 femoral access sites were closed with the fascia closure technique; 17 sites with smaller 12-F introducers were closed using other techniques. Computed tomographic angiography (CTA) was performed within 30 days, at 6 months, and at 1, 2, and 3 years. Of the 81 femoral access sites closed with the fascia closure technique, only 1 patient had persistent bleeding that required an immediate cutdown and suture repair of the deep femoral artery (99.0% technical success rate). In the immediate postoperative period, 5 patients required additional interventions for bleeding (n = 2), occlusion (n = 2), or a pseudoaneurysm [92.6% 30-day technical success]. At 30 days, 11 (13.9%) of 79 access sites had pseudoaneurysms, all of which resolved within a year; none required a secondary intervention. Later surveillance scans did not detect pseudoaneurysms. The fascia closure technique during EVAR is safe and has few complications. The low frequencies of pseudoaneurysms and other access site complications make the femoral closure technique a durable alternative.


HIV testing and preventive services accessibility among men who have sex with men at high risk of HIV infection in Beijing, China.
The HIV epidemic among men who have sex with men (MSM) has been increasing at an alarming rate in most areas of China in recent years. Many Chinese MSM still lack sufficient access to HIV prevention services, despite ongoing scale-up of comprehensive HIV testing and intervention services. The purpose of this study was to investigate utilization of HIV testing and prevention services, and related factors that influence the MSM people to access HIV test or other services to prevent HIV among MSM in Beijing, China.Three successive cross-sectional surveys of MSM were conducted in Beijing from September 2009 to January 2010, September 2010 to January 2011, and September 2011 to January 2012. Demographic and behavioral data were collected and analyzed. Blood samples were tested for HIV and syphilis. Three models were established to analyze factors associated with HIV testing and preventive services.Of the 1312 participants, prevalence of HIV and syphilis was 7.9% and 15.4%, respectively. Sixty-nine percent ever had an HIV test, 56.2%, 78.7%, and 46.1% received HIV test, free condom/lubricants, and sexually transmitted infection services in the past 12 months (P12M), respectively. MSM with larger social networks and who knew someone infected with HIV were more likely to receive HIV testing and preventive services; lower degrees of stigma and discriminatory attitudes toward HIV/AIDS were positively associated with having an HIV test, whereas unprotected anal intercourse in the past 6 months (P6M) was associated with less preventive services participation. The most reported barriers to HIV testing were fear of testing HIV positive (79.3%) and perceiving no risk for HIV (75.4%). Almost all participants felt that ensuring confidentiality would encourage more MSM to have an HIV test. The two main reasons for not seeking HIV test was not knowing where to go for a test (63.2%) and perceiving low risk of HIV infection (55.1%).Given a high prevalence of HIV, syphilis, and risky behaviors and a relatively low HIV testing rate among MSM in Beijing, more efforts are urgently needed to address barriers to HIV testing and improve accessibility of prevention services.


An analytical coarse-graining method which preserves the free energy, structural correlations, and thermodynamic state of polymer melts from the atomistic to the mesoscale.
Structural and thermodynamic consistency of coarse-graining models across multiple length scales is essential for the predictive role of multi-scale modeling and molecular dynamic simulations that use mesoscale descriptions. Our approach is a coarse-grained model based on integral equation theory, which can represent polymer chains at variable levels of chemical details. The model is analytical and depends on molecular and thermodynamic parameters of the system under study, as well as on the direct correlation function in the k → 0 limit, c0. A numerical solution to the PRISM integral equations is used to determine c0, by adjusting the value of the effective hard sphere diameter, dHS, to agree with the predicted equation of state. This single quantity parameterizes the coarse-grained potential, which is used to perform mesoscale simulations that are directly compared with atomistic-level simulations of the same system. We test our coarse-graining formalism by comparing structural correlations, isothermal compressibility, equation of state, Helmholtz and Gibbs free energies, and potential energy and entropy using both united atom and coarse-grained descriptions. We find quantitative agreement between the analytical formalism for the thermodynamic properties, and the results of Molecular Dynamics simulations, independent of the chosen level of representation. In the mesoscale description, the potential energy of the soft-particle interaction becomes a free energy in the coarse-grained coordinates which preserves the excess free energy from an ideal gas across all levels of description. The structural consistency between the united-atom and mesoscale descriptions means the relative entropy between descriptions has been minimized without any variational optimization parameters. The approach is general and applicable to any polymeric system in different thermodynamic conditions.


mTOR activation is required for the anti-alcohol effect of ketamine, but not memantine, in alcohol-preferring rats.
Glutamate NMDA receptors mediate many molecular and behavioral effects of alcohol, and they play a key role in the development of excessive drinking. Uncompetitive NMDA receptor antagonists may, therefore, have therapeutic potential for alcoholism. The first aim was to compare the effects of the NMDA antagonists memantine and ketamine on ethanol and saccharin drinking in alcohol-preferring rats. The second aim was to determine whether the effects of the two NMDA receptor antagonists were mediated by the mammalian target of rapamycin (mTOR). TSRI Sardinian alcohol-preferring rats were allowed to self-administer either 10% w/v ethanol or 0.08% w/v saccharin, and water. Operant responding and motor activity were assessed following administration of either memantine (0-10mg/kg) or ketamine (0-20mg/kg). Finally, ethanol self-administration was assessed in rats administered with either memantine or ketamine but pretreated with the mTOR inhibitor rapamycin (2.5mg/kg). The uncompetitive NMDA receptor antagonists memantine and ketamine dose-dependently reduced ethanol drinking in alcohol-preferring rats; while memantine had a preferential effect on alcohol over saccharin, ketamine reduced responding for both solutions. Neither antagonist induced malaise, as shown by the lack of effect on water intake and motor activity. The mTOR inhibitor rapamycin blocked the effects of ketamine, but not those of memantine. Memantine and ketamine both reduce alcohol drinking in alcohol-preferring rats, but only memantine is selective for alcohol. The effects of ketamine, but not memantine, are mediated by mTOR. The results support the therapeutic potential of uncompetitive NMDA receptor antagonists, especially memantine, in alcohol addiction.


Modeling the specific CD4+ T cell response against a tumor neoantigen.
The antitumor activity of CD4(+) T cells is increasingly acknowledged in both humans and mice. The involved mechanisms have been mostly studied using transplanted tumor mouse systems. In these models, many tumor cells die at the time of implantation leading to the release of Ag in an inflammatory context contrasting with the slow and nondestructive growth of early-stage human tumors. In this study, we show that the presentation of a MHC class II-restricted model Ag (male, DBY) released by dying tumor cells may last more than 4 wk. The duration of Ag presentation varies according to the way the cells are killed before implantation. To avoid this artifactual early priming of the host precluding the study of the interactions between the immune system and tumors at the steady state, we generated a cell line expressing the DBY Ag in an inducible manner. Ag expression can be efficiently induced in vivo several days after tumor implantation. We show that the Ag reaches the lymph node and activates naive CD4(+) T cells to proliferate and recirculate. We did not observe de novo induction of tumor-specific regulatory T cells. However, we observed Th1/Th17 effector cells in the tumor draining lymph node and tumors. Thus, when a neoantigen appears in established tumors, the immune system is not ignorant and naive CD4(+) T cells are not tolerized. This opens up the possibility of therapeutic vaccines improving the immune response toward tumor-specific neoantigens.


Essential sequence of influenza B virus hemagglutinin DNA to provide protection against lethal homologous viral infection.
Hemagglutinin (HA) is the main surface glycoprotein of influenza B virus. The B/Ibaraki/2/85 virus HA gene is 1758 bp in length, including signal peptide sequence, HA1 sequence, and HA2 sequence. We previously proved that B/Ibaraki/2/85 HA DNA induced immune response and provided effective protection in mice against challenge with homologous virus. In this study, a series of recombinant plasmids encoding truncated HA gene were constructed by PCR. BALB/c mice were immunized with the plasmids and challenged with a lethal dose of homologous virus. The essential sequence of HA DNA against influenza virus was explored by evaluation of survival rate, lung virus titer, bodyweight change, and serum anti-HA antibody titer of mice. The result showed that serial deletion did not deprive HA DNA of its protective ability until 885 nucleotides (295 amino acids) at 3'-terminal or 9 nucleotides of the signal peptide sequence at 5'-terminal were deleted. When the signal peptide sequence was kept intact and the 5'-terminal deletion started at the beginning of the HA1 sequence, deletion of 51 nucleotides (17 amino acids) made HA DNA lose its protective ability. This suggests that the sequence nt94-876 of B/Ibaraki/2/85 virus HA DNA played an important role in protection against infection.


Brain Structure and Degeneration Staging in Friedreich Ataxia: Magnetic Resonance Imaging Volumetrics from the ENIGMA-Ataxia Working Group.
Friedreich ataxia (FRDA) is an inherited neurological disease defined by progressive movement incoordination. We undertook a comprehensive characterization of the spatial profile and progressive evolution of structural brain abnormalities in people with FRDA. A coordinated international analysis of regional brain volume using magnetic resonance imaging data charted the whole-brain profile, interindividual variability, and temporal staging of structural brain differences in 248 individuals with FRDA and 262 healthy controls. The brainstem, dentate nucleus region, and superior and inferior cerebellar peduncles showed the greatest reductions in volume relative to controls (Cohen d = 1.5-2.6). Cerebellar gray matter alterations were most pronounced in lobules I-VI (d = 0.8), whereas cerebral differences occurred most prominently in precentral gyri (d = 0.6) and corticospinal tracts (d = 1.4). Earlier onset age predicted less volume in the motor cerebellum (rmax = 0.35) and peduncles (rmax = 0.36). Disease duration and severity correlated with volume deficits in the dentate nucleus region, brainstem, and superior/inferior cerebellar peduncles (rmax = -0.49); subgrouping showed these to be robust and early features of FRDA, and strong candidates for further biomarker validation. Cerebral white matter abnormalities, particularly in corticospinal pathways, emerge as intermediate disease features. Cerebellar and cerebral gray matter loss, principally targeting motor and sensory systems, preferentially manifests later in the disease course. FRDA is defined by an evolving spatial profile of neuroanatomical changes beyond primary pathology in the cerebellum and spinal cord, in line with its progressive clinical course. The design, interpretation, and generalization of research studies and clinical trials must consider neuroanatomical staging and associated interindividual variability in brain measures. ANN NEUROL 2021;90:570-583.


Multi-Radiologist User Study for Artificial Intelligence-Guided Grading of COVID-19 Lung Disease Severity on Chest Radiographs.
Radiographic findings of COVID-19 pneumonia can be used for patient risk stratification; however, radiologist reporting of disease severity is inconsistent on chest radiographs (CXRs). We aimed to see if an artificial intelligence (AI) system could help improve radiologist interrater agreement. We performed a retrospective multi-radiologist user study to evaluate the impact of an AI system, the PXS score model, on the grading of categorical COVID-19 lung disease severity on 154 chest radiographs into four ordinal grades (normal/minimal, mild, moderate, and severe). Four radiologists (two thoracic and two emergency radiologists) independently interpreted 154 CXRs from 154 unique patients with COVID-19 hospitalized at a large academic center, before and after using the AI system (median washout time interval was 16 days). Three different thoracic radiologists assessed the same 154 CXRs using an updated version of the AI system trained on more imaging data. Radiologist interrater agreement was evaluated using Cohen and Fleiss kappa where appropriate. The lung disease severity categories were associated with clinical outcomes using a previously published outcomes dataset using Fisher's exact test and Chi-square test for trend. Use of the AI system improved radiologist interrater agreement (Fleiss κ = 0.40 to 0.66, before and after use of the system). The Fleiss κ for three radiologists using the updated AI system was 0.74. Severity categories were significantly associated with subsequent intubation or death within 3 days. An AI system used at the time of CXR study interpretation can improve the interrater agreement of radiologists.


Efficacy of Allium sativum oil to alleviate tebuconazol-induced oxidative stress in the liver of adult rats.
The present study focused on the protective efficacy of Allium sativum oil (ASO) against tebuconazol (TEB)-induced oxidative stress in the liver of adult rats. Thirty-two rats were randomly divided into four groups of eight each: group I served as control rats, group II was treated with TEB (100 mg/kg bw), group III received ASO (5ml/kg bw). The animals of group IV were treated with TEB and ASO, during 4 weeks. The obtained results showed that TEB induced a significant change of some hematological parameters, including red blood cells (RBC), haemoglobin content (Hb), haematocrit (Ht), white blood cells (WBC) and platelet (Plt) compared to the control group. Moreover, while the total cholesterol levels and the activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH) and γ-Glutamyltranspeptidase (γGT) significantly increased due to TEB administration, the concentrations of plasma total protein, albumin and triglyceride considerably decreased. Furthermore, the exposure to TEB significantly increased the malondialdehyde (MDA), protein carbonyl (PCO) and advanced oxidation protein products (AOPP) levels and decreased glutathione peroxidase (GPx), superoxide dismutase (SOD), catalase (CAT) and glutathione-S-transferase (GST) activities in the hepatic tissues. The results were confirmed by the histological impairments. Besides, the co-administration of ASO improved the status of all studied parameters. Therefore, our investigation revealed that ASO had protective effects against TEB-induced liver injury, which could be attributed to its phenolic compounds.


Presence of intestinal Mycobacterium avium subspecies paratuberculosis (MAP) DNA is not associated with altered MMP expression in ulcerative colitis.
Mycobacterium avium subspecies paratuberculosis (MAP) is suspected to be a causative agent in human Crohn's disease (CD). Recent evidence suggests that pathogenic mycobacteria and MAP can induce the expression of Matrix Metalloproteinases (MMP), which are the main proteases in the pathogenesis of mucosal ulcerations in inflammatory bowel disease (IBD). Within this study we assessed the prevalence of intestinal MAP specific DNA in patients with Crohn's disease, ulcerative colitis (UC), and healthy controls. We further analysed regulation patterns of MMPs in mucosal tissues of UC patients with and without intestinal MAP DNA detection. Colonic biopsy samples were obtained from 63 Norwegian and German IBD patients and 21 healthy controls. RNA was quantified by quantitative real-time polymerase chain reaction (PCR) to study MMP gene expression in both pathological and healthy mucosal specimens. The presence of MAP DNA in colonic mucosa was examined using MAP specific PCR. MAP DNA was detected in 20% of UC patients and 33% of healthy controls but only in 7% of patients with CD. UC patients treated with corticosteroids exhibited a significantly increased frequency of intestinal MAP DNA compared to those not receiving corticosteroids. Expression of MMP-1, -2, -7, -9, -13, -19, -28 and TNF-α did not differ between UC patients with presence of intestinal MAP DNA compared to those without. MMP-2, MMP-9 and MMP-13 were significantly decreased in UC patients receiving corticosteroids. The presence of intestinal MAP specific DNA is not associated with altered MMP expression in UC in vivo. Corticosteroids are associated with increased detection of intestinal MAP DNA and decreased expression of certain MMPs. Frequent detection of MAP DNA in healthy controls might be attributable to the wide environmental distribution of MAP and its presence in the food-chain.


Relapsing polychondritis presenting with sero-negative limbic encephalitis.
The presented case highlights a rare instance of relapsing polychondritis (RP) manifesting as seronegative limbic encephalitis, an uncommon neurological complication. A 70-year-old female patient with a history of RP-related inflammation, along with neuropsychiatric symptoms, was diagnosed through multidisciplinary collaboration. Swift administration of steroid therapy, followed by azathioprine, led to remarkable physical and cognitive recovery. This case emphasises the importance of a multidisciplinary approach in diagnosing and treating complex autoimmune disorders with neurological manifestations.


Effects of secukinumab on serum adipocytokines: preliminary data.
Psoriatic arthritis (PsA) is a chronic inflammatory disease that affects joints, connective tissues and the axial skeleton. Metabolic syndrome is an independent risk factor for psoriasis (Pso) development and is associated with more severe forms of Pso. Adipocytokines are secreted by white adipose tissue and are thought to link obesity with the development of metabolic and cardiovascular diseases. Secukinumab is a new monoclonal antibody with a different mechanism of action. This antibody selectively binds to and neutralizes interleukin-17 (IL-17) and it has shown efficacy in the treatment of PsA. The aim of this study was to evaluate the possible interferences of secukinumab on different adipocytokines. We enrolled 28 patients with PsA, classified with the CASPAR criteria. Serum samples were stored at baseline and then at the first, the third and the sixth month of therapy. Resistin, chemerin, adiponectin and C-reactive protein (CRP) were dosed. When tested globally, none of the adipokine tested showed any statistically significant variation. However, when the male group was tested, both resistin and chemerin at M6 showed a significant decrease from baseline. CRP did not show any variation at any time point. Our study demonstrated that treatment with secukinumab has little influence on the levels of adipokines tested within the first six months of treatment even though it might exert different influence between males and females from a metabolic perspective. Further studies with greater numbers of patients are needed to determine whether these preliminary results have clinical relevance.


Pivoting to teleconsultation for paediatric ophthalmology and strabismus: Our experience during COVID-19 times.
The objective is to analyse and report the data of teleconsultations provided to paediatric ophthalmology and strabismus patients during COVID-19 times and to elaborate our experience for guiding future teleconsultation practices to General, paediatric Ophthalmologists and Strabismologists. Retrospective analysis of electronic medical record data of teleconsultations provided in the department of Strabismus, Paediatric and Neuro-ophthalmology was done. Patients with optic nerve related disorders were excluded. Study period was one month. Statistical analysis of collected data was done using Microsoft excel. A total of 198 patients were provided teleconsultations (an average of seven teleconsultations/day). The final analysis included 161 patients after excluding optic nerve related disorders. The median age was seven years. We had a near equal gender distribution (53% males and 47% females) of whom a third were new cases. Video calling was used in 14%, review of clinical photos shared was used in 53%. Rest of the 33% were given telephonic advice. Allergic conjunctivitis (14%), pseudophakia (9%), strabismus (12%), status post strabismus surgery (8%), cranial nerve palsies (11%) were common diagnoses. 19% (n = 30) were advised/needed visit in emergency services on same or next day. Our experience of teleconsultation during COVID-19 times for paediatric ophthalmology and strabismus patients was very encouraging. Pivoting teleconsultation platform can provide primary eye care to most of the patients and work as essential forward triage for rest. As we continue to further explore the currently available avenues in multimedia such as video conferencing and web/mobile based applications, we believe that tele-ophthalmology platform can provide a reliable service in patient care.


Integrin signaling potentiates transforming growth factor-beta 1 (TGF-β1) dependent down-regulation of E-Cadherin expression - Important implications for epithelial to mesenchymal transition (EMT) in renal cell carcinoma.
Signal transduction through the transforming growth factor-beta 1 (TGF-β1) pathway affects epithelial to mesenchymal transition (EMT), partly by modulation of E-Cadherin expression. The concurrent impact of extracellular matrix driven regulation of integrin signaling on EMT has not been well characterized. We assessed the cumulative effect and molecular mechanisms of TGF-β1 and integrin signal transduction on E-Cadherin in a renal cell cancer (RCC) model. Stimulation of RCC cells with TGF-β1 demonstrated a three-fold increased expression of integrin αv. A ligand of integrin αv-β3, (cyclopentapeptide containing Arginyl-Glycyl-Aspartic acid motif, RGD), was used to mimic integrin signaling. Treatment of cells with RGD and TGF-β1 demonstrated significantly greater E-cadherin depletion than either ligand alone. This cooperative action on E-Cadherin expression is regulated by transcription factor Snai1 and is followed on a cellular level by increased cellular mobility as evidenced in a wound healing assay. Subsequent silencing of potential downstream mediators of the cumulative action of RGD and TGF-β1 was carried out by small interfering RNA transfection and confirmed by Western blotting and/or RT-PCR. SiRNA mediated silencing of FAK and PINCH1 independently abrogated the cumulative effect of RGD and TGF-β1 on E-Cadherin expression. We have identified a novel mechanism through which extracellular matrix event transduction by integrins further augments TGF-β1 related effects on EMT. Molecular machinery involved in the integrin αv-TGF-β1 interplay may represent a therapeutic target in RCC.


Plastic microchip electrophoresis for analysis of PCR products of hepatitis C virus.
Electrophoresis on polymeric rather than glass microstructures is a promising separation method for analytical chemistry. Assays on such devices need to be explored to allow assessment of their utility for the clinical laboratory. We compared capillary and plastic microchip electrophoresis for clinical post-PCR analysis of hepatitis C virus (HCV). For capillary electrophoresis (CE), we used a separation medium composed of 10 g/L hydroxypropyl methyl cellulose in Tris-borate-EDTA buffer and 10 micromol/L intercalating dye. For microchip electrophoresis, the HCV assay established on the fused silica tubing was transferred to the untreated polymethylmethacrylate microchip with minimum modifications. CE resolved the 145-bp amplicon of HCV in 15 min. The confidence interval of the migration time was <3.2%. The same HCV amplicon was resolved by microchip electrophoresis in <1.5 min with the confidence interval of the migration time <1.3%. The polymer microchip, with advantages that include fast processing time, simple operation, and disposable use, holds great potential for clinical analysis.


Mechanistic studies on the biosorption of Pb(II) by Pseudomonas aeruginosa.
The biosorption of Pb(II) ions from aqueous solution has been studied using both the intact and thermolyzed cells of Pseudomonas aeruginosa. Further, the role of the major cell wall components, namely DNA, protein, polysaccharide, and lipid, in Pb(II) binding has been assessed using an enzymatic treatment method. The Pb(II) bioremediation capability of P. aeruginosa cells has been investigated by varying the parameters of pH, time of interaction, amount of biomass, and concentration of Pb(II). The complete bioremoval of Pb(II) using intact cells has been achieved for an initial Pb(II) concentration of 12.4 mg L-1 at pH 6.2 and temperature 29 ± 1 °C. The biosorption isotherm follows Langmuirian behavior with a Gibbs free energy of -30.7 kJ mol-1, indicative of chemisorption. The biosorption kinetics is consistent with a pseudo-second-order model. The possible Pb(II) binding mechanisms of P. aeruginosa cells are discussed based on characterization using zeta potential measurements, Fourier transform infra-red spectroscopy, and energy dispersive X-ray spectroscopy. The results confirm that among the major cell wall components studied, polysaccharide shows the highest contribution towards Pb(II) binding, followed by DNA, lipid, and protein. Similar studies using thermolyzed cells show higher Pb(II) uptake compared to the intact cells both before and after enzymatic treatment.


Studies on osteoporoses. XI. Effects of a methylxanthine derivative. A preliminary report.
Heparin (500 U/kg s.c. B.I.D.) induced significant osteoporosis in C3H/St(Ha) female mice after 3 months of treatment. Pentoxifylline (12 mg/kg i.m. B.I.D.) prevented this experimental osteoporosis. Osteoporosis was measured by in vivo neutron activation analysis and results were confirmed by atomic absorption spectroscopy. Pentoxifylline (0.1-100 microgram/ml) increased calcium uptake and cAMP production in osteoblast-like bone cells isolated from fetal Sprague-Dawley rats. Theoretical implications for osteoblast control of bone resorption are discussed.


Reciprocal expression of ERalpha and ERbeta is associated with estrogen-mediated modulation of intestinal tumorigenesis.
Menopausal hormone replacement therapy has been widely used to alleviate the symptoms of menopause and to decrease the detrimental effects of ovarian hormone loss on bone density and cardiovascular health. Multiple studies of colorectal cancer epidemiology also support a role for hormone replacement therapy in prevention of colorectal cancer. We studied the effect of ovariectomy and estrogen replacement on tumor formation in C57BL/6J-Min/+ (Min/+) mice, animals that bear a germline mutation in murine Apc. These mice develop multiple intestinal tumors that show loss of wild-type Apc protein. After ovariectomy, intestinal adenomas in Min/+ mice increased by 77% (P = 0.0004). Ovariectomized Min/+ mice that were treated with a replacement dose of 17beta-estradiol had the same number of tumors as Min/+ mice that were neither castrated nor treated with estrogen replacement (P = 0.85). Examination of estrogen receptor (ER) levels in intestinal tissue by immunoblot showed changes in relative expression levels of ERalpha and ERbeta, with highest ERalpha and lowest ERbeta expression in the normal-appearing intestine of Min/+ mice, and lowest ERalpha and highest ERbeta expression in the enterocytes of animals that received 17beta-estradiol. These results suggest that endogenous estrogens protect against Apc-associated tumor formation and that tumor prevention by 17beta-estradiol is associated with an increase in ERbeta and a decrease in ERalpha expression in the target tissue.


Adapting Mobile and Wearable Technology to Provide Support and Monitoring in Rehabilitation for Dementia: Feasibility Case Series.
Mobile and wearable devices are increasingly being used to support our everyday lives and track our behavior. Since daily support and behavior tracking are two core components of cognitive rehabilitation, such personal devices could be employed in rehabilitation approaches aimed at improving independence and engagement among people with dementia. The aim of this work was to investigate the feasibility of using smartphones and smartwatches to augment rehabilitation by providing adaptable, personalized support and objective, continuous measures of mobility and activity behavior. A feasibility study comprising 6 in-depth case studies was carried out among people with early-stage dementia and their caregivers. Participants used a smartphone and smartwatch for 8 weeks for personalized support and followed goals for quality of life. Data were collected from device sensors and logs, mobile self-reports, assessments, weekly phone calls, and interviews. This data were analyzed to evaluate the utility of sensor data generated by devices used by people with dementia in an everyday life context; this was done to compare objective measures with subjective reports of mobility and activity and to examine technology acceptance focusing on usefulness and health efficacy. Adequate sensor data was generated to reveal behavioral patterns, even for minimal device use. Objective mobility and activity measures reflecting fluctuations in participants' self-reported behavior, especially when combined, may be advantageous in revealing gradual trends and could provide detailed insights regarding goal attainment ratings. Personalized support benefited all participants to varying degrees by addressing functional, memory, safety, and psychosocial needs. A total of 4 of 6 (67%) participants felt motivated to be active by tracking their step count. One participant described a highly positive impact on mobility, anxiety, mood, and caregiver burden, mainly as a result of navigation support and location-tracking tools. Smartphones and wearables could provide beneficial and pervasive support and monitoring for rehabilitation among people with dementia. These results substantiate the need for further investigation on a larger scale, especially considering the inevitable presence of mobile and wearable technology in our everyday lives for years to come.


The Anti-Inflammatory Actions of Auricular Point Acupressure for Chronic Low Back Pain.
Background. Auricular point acupressure (APA) is a promising treatment for pain management. Few studies have investigated the physiological mechanisms of APA analgesics. Method. In this pilot randomized clinical trial (RCT), a 4-week APA treatment was used to manage chronic low back pain (CLBP). Sixty-one participants were randomized into a real APA group (n = 32) or a sham APA group (n = 29). Blood samples, pain intensity, and physical function were collected at baseline and after 4 weeks of treatment. Results. Subjects in the real APA group reported a 56% reduction of pain intensity and a 26% improvement in physical function. Serum blood samples showed (1) a decrease in IL-1β, IL-2, IL-6, and calcitonin gene-related peptide [CGRP] and (2) an increase in IL-4. In contrast, subjects in the sham APA group (1) reported a 9% reduction in pain and a 2% improvement in physical function and (2) exhibited minimal changes of inflammatory cytokines and neuropeptides. Statistically significant differences in IL-4 and CGRP expression between the real and sham APA groups were verified. Conclusion. These findings suggest that APA treatment affects pain intensity through modulation of the immune system, as reflected by APA-induced changes in serum inflammatory cytokine and neuropeptide levels.


Factors associated with poor health-related quality of life of patients with cirrhosis.
Increasing interest is being given to health-related quality of life in chronic diseases. In cirrhosis, both physical functioning and mental well-being may be altered, but no study has investigated factors associated with a poor perceived health status. We measured quality of life by Short Form-36 and Nottingham Health Profile questionnaires in 544 patients with cirrhosis. Data were compared with age- and gender-adjusted values of 2 random samples of the Italian population (more than 2000 subjects). Factors associated with poor perceived health status were identified by logistic regression. All domains of health-related quality of life, except pain, were altered in cirrhosis (by 9%-42%), mainly in younger patients. There were minor differences in relation to gender, whereas etiology had no effects. Severity of disease (Child-Pugh score) and, above all, muscle cramps were the factors most closely associated with poor health status perception. Self-rating of disease progression was associated with ascites and pruritus, whereas previous variceal sclerotherapy and the use of disaccharides had a protective effect. Most areas of daily life were affected by perceived health problems; this was mainly true for paid employment and sex life in men and home life and social life in women. Quality of life is variably impaired in cirrhosis, also in uncomplicated patients. Non-life-threatening symptoms, such as muscle cramps, are of major concern. These data are the basis for longitudinal studies measuring the effects of therapy and procedures on patient-derived health outcomes.


ANTIMICROBIAL ACTIVITIES OF CHITOSAN NANOPARTICLES PREPARED-FROM LUCILA CUPRINA MAGGOTS (DIPTERA: CALLIPHORIDAE).
Chitosan nanoparticles Were studied as antimicrobial agent. The antibacterial activity of chitosan nanoparticles were investigated against three Gram-negative bacteria; Escherichia coli, Pseudomonas aeruginosa and Salmonella typhi, and three Gram-positive bacteria; Staphylococcus aureus, Enterococcus faecalis and Streptococcus pyogenes. The 'antifungal activity were examined against three fungi; Geotrichum candidum, Candida krusei and Candida parapsilosis. The antiviral activities were tested against three viruses; Rift Valley Fever (RVFV), Herpes simplex-I (HSV-1) and Coxsackie viruses. Chitosan nanoparticles were inhibited all bacteria and fungi except E. faecalis seemed to be resistant strain. Infectivity titers of all viruses were reduced by chitosan nanoparticles, which are a natural antimicrobial agent.


Clinical significance of IgA rheumatoid factor subclasses in rheumatoid arthritis.
IgA rheumatoid factor (RF) is associated with greater disease activity and radiological progression in rheumatoid arthritis (RA). We examined whether measuring IgA RF subclasses gives additional clinically relevant information. Total IgA RF plus IgA1 RF and IgA2 RF subclasses were estimated by ELISA using rabbit IgG as antigen in 144 patients with established RA. Disease activity was assessed by Disease Activity Score and Health Assessment Questionnaire; the acute phase response was assessed by C-reactive protein levels and joint damage by the Larsen score. Fifty percent of patients had elevated total IgA RF, 60% elevated IgA1 RF, and 50% elevated IgA2 RF. There were significant correlations between total IgA RF and both IgA RF subclasses (p < 0.0001). Measures of disease activity, the acute phase response, and joint damage were all significantly higher (p < 0.001 in each case) in patients with elevated total IgA RF and both IgA RF subclasses compared with RF negative cases. Disease duration influenced the relationship of IgA RF subclasses to joint damage. The Larsen score was only significantly higher in late disease (duration 5 years or more) in both IgA1 RF and IgA2 RF positive patients. We found IgA RF positive patients have more aggressive RA than negative cases. Measuring IgA1 RF and IgA2 RF subclasses did not give more information about clinical status than measuring total IgA RF alone.


2023 WSES guidelines for the prevention, detection, and management of iatrogenic urinary tract injuries (IUTIs) during emergency digestive surgery.
Iatrogenic urinary tract injury (IUTI) is a severe complication of emergency digestive surgery. It can lead to increased postoperative morbidity and mortality and have a long-term impact on the quality of life. The reported incidence of IUTIs varies greatly among the studies, ranging from 0.3 to 1.5%. Given the high volume of emergency digestive surgery performed worldwide, there is a need for well-defined and effective strategies to prevent and manage IUTIs. Currently, there is a lack of consensus regarding the prevention, detection, and management of IUTIs in the emergency setting. The present guidelines, promoted by the World Society of Emergency Surgery (WSES), were developed following a systematic review of the literature and an international expert panel discussion. The primary aim of these WSES guidelines is to provide evidence-based recommendations to support clinicians and surgeons in the prevention, detection, and management of IUTIs during emergency digestive surgery. The following key aspects were considered: (1) effectiveness of preventive interventions for IUTIs during emergency digestive surgery; (2) intra-operative detection of IUTIs and appropriate management strategies; (3) postoperative detection of IUTIs and appropriate management strategies and timing; and (4) effectiveness of antibiotic therapy (including type and duration) in case of IUTIs.


A comparative analysis of mitochondrial genomes in Coleoptera (Arthropoda: Insecta) and genome descriptions of six new beetles.
Coleoptera is the most diverse group of insects with over 360,000 described species divided into four suborders: Adephaga, Archostemata, Myxophaga, and Polyphaga. In this study, we present six new complete mitochondrial genome (mtgenome) descriptions, including a representative of each suborder, and analyze the evolution of mtgenomes from a comparative framework using all available coleopteran mtgenomes. We propose a modification of atypical cox1 start codons based on sequence alignment to better reflect the conservation observed across species as well as findings of TTG start codons in other genes. We also analyze tRNA-Ser(AGN) anticodons, usually GCU in arthropods, and report a conserved UCU anticodon as a possible synapomorphy across Polyphaga. We further analyze the secondary structure of tRNA-Ser(AGN) and present a consensus structure and an updated covariance model that allows tRNAscan-SE (via the COVE software package) to locate and fold these atypical tRNAs with much greater consistency. We also report secondary structure predictions for both rRNA genes based on conserved stems. All six species of beetle have the same gene order as the ancestral insect. We report noncoding DNA regions, including a small gap region of about 20 bp between tRNA-Ser(UCN) and nad1 that is present in all six genomes, and present results of a base composition analysis.


Electronic Cigarettes: Questions in the Mist.
In many parts of the world, electronic cigarettes (ECs) are marketed as a tool to assist users in attempts to quit smoking and are perceived to be a safer alternative to tobacco cigarettes. Numerous studies have suggested ECs may not be effective in achieving this goal and that the illusive "safety" of ECs can be enticing to consumers. The composition of the liquid solutions vaporized by these devices has not been fully disclosed and may not contain the actual advertised ingredients in the fractions reported. As this market has developed largely outside the oversight of an appropriate regulatory body, we are left to evaluate how ECs as a form of nicotine replacement therapy fit in the existing regulatory framework of conventional cigarettes and what the role of nicotine is in our society.


Systematic symptom management in the IMPACT Consortium: rationale and design for 3 effectiveness-implementation trials.
Cancer and its treatment produce deleterious symptoms across the phases of care. Poorly controlled symptoms negatively affect quality of life and result in increased health-care needs and hospitalization. The Improving the Management of symPtoms during And following Cancer Treatment (IMPACT) Consortium was created to develop 3 large-scale, systematic symptom management systems, deployed through electronic health record platforms, and to test them in pragmatic, randomized, hybrid effectiveness and implementation trials. Here, we describe the IMPACT Consortium's conceptual framework, its organizational components, and plans for evaluation. The study designs and lessons learned are highlighted in the context of disruptions related to the COVID-19 pandemic.


Efficacy and safety of subcutaneous and intravenous rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisone in first-line diffuse large B-cell lymphoma: the randomized MabEase study.
Intravenous rituximab plus chemotherapy is standard treatment for diffuse large B-cell lymphoma. A subcutaneous formulation of rituximab is expected to simplify and shorten drug preparation and administration, and to reduce treatment burden. MabEase (clinicaltrials.gov Identifier: 01649856) examined efficacy, safety and patient satisfaction with subcutaneous rituximab plus chemotherapy in treatment-naïve patients with diffuse large B-cell lymphoma. Patients were randomized 2:1 to subcutaneous rituximab (intravenous 375 mg/m2 cycle 1; subcutaneous 1,400 mg cycles 2-8) or intravenous rituximab (375 mg/m2 cycles 1-8) plus cyclophosphamide, doxorubicin, vincristine, and prednisone every 14 or 21 days. The primary endpoint was investigator-assessed complete response/unconfirmed complete response. Secondary endpoints included safety, treatment satisfaction (Cancer Treatment Satisfaction Questionnaire and Rituximab Administration Satisfaction Questionnaire), time savings, and survival. Of 576 randomized patients, 572 (378 subcutaneous; 194 intravenous) received treatment. End of induction complete response/unconfirmed complete response rates were 50.6% (subcutaneous) and 42.4% (intravenous). After a median 35 months, median overall, event-free and progression-free survivals were not reached. Grade ≥3 adverse events (subcutaneous 58.3%; intravenous 54.3%) and administration-related adverse events (both groups 21%) were similar between arms. Injection-site reactions were more common with subcutaneous injections (5.7% versus 0%, respectively). Rituximab Administration Satisfaction Questionnaire scores for 'impact on activities of daily living', 'convenience', and 'satisfaction' were improved with subcutaneous versus intravenous injections; Cancer Therapy Satisfaction Questionnaire scores were similar between arms. Median administration time (6 minutes vs 2.6 to 3.0 hours), chair/bed and overall hospital times were shorter with subcutaneous versus intravenous rituximab. Overall, subcutaneous and intravenous rituximab had similar efficacy and safety, with improved patient satisfaction and time savings.


Natural allelic variation identifies new genes in the Arabidopsis circadian system.
We have analysed the circadian rhythm of Arabidopsis thaliana leaf movements in the accession Cvi from the Cape Verde Islands, and in the commonly used laboratory strains Columbia (Col) and Landsberg (erecta) (Ler), which originated in Northern Europe. The parental lines have similar rhythmic periods, but the progeny of crosses among them reveal extensive variation for this trait. An analysis of 48 Ler/Cvi recombinant inbred lines (RILs) and a further 30 Ler/Col RILs allowed us to locate four putative quantitative trait loci (QTLs) that control the period of the circadian clock. Near-isogenic lines (NILs) that contain a QTL in a small, defined chromo- somal region allowed us to confirm the phenotypic effect and to map the positions of three period QTLs, designated ESPRESSO, NON TROPPO and RALENTANDO. Quantitative trait loci at the locations of RALENTANDO and of a fourth QTL, ANDANTE, were identified in both Ler/Cvi and Ler/Col RIL populations. Some QTLs for circadian period are closely linked to loci that control flowering time, including FLC. We show that flc mutations shorten the circadian period such that the known allelic variation in the MADS-box gene FLC can account for the ANDANTE QTL. The QTLs ESPRESSO and RALENTANDO identify new genes that regulate the Arabidopsis circadian system in nature, one of which may be the flowering-time gene GIGANTEA.


¹⁵N, ¹³C and ¹H resonance assignments and secondary structure determination of a variable heavy domain of a heavy chain antibody.
Heavy chain antibodies differ in structure to conventional antibodies lacking both the light chain and the first heavy chain constant domain (CH1). Characteristics of the antigen-binding variable heavy domain of the heavy chain antibody (VHH) including the smaller size, high solubility and stability make them an attractive alternative to more traditional antibody fragments for detailed NMR-based structural analysis. Here we report essentially complete backbone and side chain (15)N, (13)C and (1)H assignments for a free VHH. Analysis of the backbone chemical shift data obtained indicates that the VHH is comprised predominantly of β-sheets corresponding to nearly 60% of the protein backbone.


Scanning electron microscopic study of the tongue in the Oriental scops owl (Otus scops).
The dorsal lingual surface of an adult owl (Otus scops) was examined by scanning electron microscopy. The tongue of the adult owl was about 1 cm long. Three parts were distinguished in the dorsal surface of the tongue: the apex, the body and the root of the tongue. The conical region between the lingual apex and lingual root was very wide area. The conical papillae of the lingual body were inclined toward the posterior of the tongue. At low magnification of scanning electron microscopy, the desquamated cells were observed in the entire dorsal surface of the lingual apex. The connective tissue cores of the epithelium of the lingual apex showed the rod-shaped protrusions. The border between the lingual apex and body was clear and the small conical papillae were observed in the lingual body. The small and large conical papillae were observed on the lingual body. The many openings of the lingual glands existed in the lingual body and lingual root.


Lipid profiles in adolescent Filipinos: relation to birth weight and maternal energy status during pregnancy.
The finding that persons with low birth weight have a higher cardiovascular disease (CVD) risk than do persons with higher birth weight remains poorly understood. We tested the hypothesis that maternal arm fat area (MAFA) in the third trimester of pregnancy and birth weight of offspring are inversely related to the offspring's risk of CVD. In a 1-y birth cohort study (1983-1984), 296 male and 307 female offspring were followed up (1998-1999) to measure their lipid profiles. Participants came from randomly selected communities of Cebu, Philippines. MAFA (log cm2) was positively associated (beta) with HDL cholesterol (0.12 log mg/dL; P < 0.01) and inversely associated with total cholesterol (-10.0 mg/dL; P < 0.10), LDL cholesterol (-13.1 mg/dL; P < 0.01), and the ratios of total to HDL cholesterol and LDL to HDL cholesterol (both P < 0.001) in males. These relations were independent of birth weight, present adiposity, energy and fat intakes, maturity, and income. Birth weight < or = 2.6 kg was associated with elevated LDL cholesterol (9.9 mg/dL; P < 0.01) and an elevated ratio of LDL to HDL cholesterol (0.22; P < 0.10) only in males. In females, MAFA related positively to total (15.5 mg/dL; P < 0.05) and LDL (11.9 mg/dL; P < 0.05) cholesterol. In this Filipino population, mothers with low energy status during pregnancy gave birth to male offspring who had a high CVD risk in adolescence, as indicated by lipid profiles. The findings in females are less consistent with the fetal origins hypothesis and suggest sex differences in the relation between fetal nutrition and postnatal lipid metabolism.


Nondilated obstructive nephropathy.
Three patients had renal failure due to obstructive nephropathy associated with processes that prevented dilatation of the collecting systems. Thus, various radiologic procedures, including renal sonography, angiography, and isotope renography, all failed to identify an obstructing process. Because of the high index of clinical suspicion, surgical exploration and nephrostomy were performed on each patient. This confirmed the presence of obstructive nephropathy and led to marked improvement of renal function in each case. When renal failure develops in a setting with a high probability of ureteral obstruction, this diagnosis should be vigorously pursued despite normal radiologic results.


Response of the Pre-Oriented Goal-Directed Attention to Usual and Unusual Distractors: A Preliminary Study.
In this study, we investigated the distraction power of the unusual and usual images on the attention of 20 healthy primary school children. Our study was different from previous ones in that the participants were asked to fix the initial position of their attention on a predefined location after being presented with unusual images as distractors. The goals were presented in locations, which were far from the attraction basin of distractors. We expected that the pre-orienting of the attention to the position of targets would reduce the attractive effect of unusual images compared to the usual ones. The percentage of correct responses and the reaction time were measured as behavioral indicators of attention performance. Results showed that using the goal-directed attention, subjects ignored both kinds of distractors nearly the same way. With regard to previous reports about more attraction towards the unusual images, it is suggested that the dynamics of the visual attention system be sensitive to the initial condition. That is, changing the initial position of the attention can lead to the decrement of the unusual images effects. However, several other possibilities such as a probable delay in processing unusual features could explain this observation, too.


Spatial scales of genetic structure and gene flow in Calochortus albus (Liliaceae).
Calochortus (Liliaceae) displays high species richness, restriction of many individual taxa to narrow ranges, geographic coherence of individual clades, and parallel adaptive radiations in different regions. Here we test the first part of a hypothesis that all of these patterns may reflect gene flow at small geographic scales. We use amplified fragment length polymorphism variation to quantify the geographic scales of spatial genetic structure and apparent gene flow in Calochortus albus, a widespread member of the genus, at Henry Coe State Park in the Coast Ranges south of San Francisco Bay. Analyses of 254 mapped individuals spaced 0.001-14.4 km apart show a highly significant decline in genetic identity with ln distance, implying a root-mean-square distance of gene flow σ of 5-43 m. STRUCTURE analysis implies the existence of 2-4 clusters over the study area, with frequent reversals among clusters over short distances (<200 m) and a relatively high frequency of admixture within individuals at most sampling sites. While the intensity of spatial genetic structure in C. albus is weak, as measured by the Sp statistic, that appears to reflect low genetic identity of adjacent plants, which might reflect repeated colonizations at small spatial scales or density-dependent mortality of individual genotypes by natural enemies. Small spatial scales of gene flow and spatial genetic structure should permit, under a variety of conditions, genetic differentiation within species at such scales, setting the stage ultimately for speciation and adaptive radiation as such scales as well.