Biologic frontiers in multiple myeloma: from biomarker identification to clinical practice.
Since the mid-1990s, the multiple myeloma treatment landscape has evolved considerably, which has led to improved patient outcomes and prolonged survival. In addition to discovering new, targeted agents or treatment regimens, the identification and validation of biomarkers has the potential to further improve patient outcomes. The International Staging System relies on a number of biochemical parameters to stratify patients into risk categories. Other biologically relevant markers that are indicative of inherited genetic variation (e.g., single-nucleotide polymorphisms) or tumor-acquired genetic events (e.g., chromosomal translocations or mutations) have been studied for their prognostic potential. In patients with high-risk cytogenetics, plasma cells (PC) undergo genetic shifts over time, which may partially explain why high-risk patients relapse and are so difficult to treat. Although novel agents have improved treatment outcomes, identification of markers that will enable clinicians to determine which treatment is most appropriate for high-risk patients following initial diagnosis represents an exciting frontier in the clinical management of multiple myeloma. Biomarkers based on quantitating PCs or factors that are secreted from them (e.g., serum free light chain) may also help to risk-stratify patients with asymptomatic multiple myeloma. Eventually, identification of novel biomarkers may lead to the creation of personalized treatment regimens that are optimized to target clonal PCs that express a specific oncogenomic profile. Although the future is exciting, validation will be necessary before these biologic and molecular beacons can inform decision-making processes in a routine clinical setting.


Rubella exposure in an obstetric clinic.
Despite a massive national rubella immunization program, rubella infection remains a public health problem. When a nurse employed in a hospital-based obstetric clinic became ill with rubella, a crisis was precipitated. The hospital staff implemented a plan to inform the exposed 151 patients and 44 employees of the potential danger to themselves and their fetuses. To identify persons at risk, a program of rubella antibody testing of contacts was instituted. In 3 distinct attempts to obtain blood specimens, patient compliance rates fell progressively from 79 to 14%. One additional infected employee was detected. A list of recommendations designed to prevent or lessen the impact of future rubella exposures in hospitals is presented.


Deciphering the Role of Selenoprotein M.
Selenocysteine (Sec), the 21st amino acid, is structurally similar to cysteine but with a sulfur to selenium replacement. This single change retains many of the chemical properties of cysteine but often with enhanced catalytic and redox activity. Incorporation of Sec into proteins is unique, requiring additional translation factors and multiple steps to insert Sec at stop (UGA) codons. These Sec-containing proteins (selenoproteins) are found in all three domains of life where they often are involved in cellular homeostasis (e.g., reducing reactive oxygen species). The essential role of selenoproteins in humans requires us to maintain appropriate levels of selenium, the precursor for Sec, in our diet. Too much selenium is also problematic due to its toxic effects. Deciphering the role of Sec in selenoproteins is challenging for many reasons, one of which is due to their complicated biosynthesis pathway. However, clever strategies are surfacing to overcome this and facilitate production of selenoproteins. Here, we focus on one of the 25 human selenoproteins, selenoprotein M (SELENOM), which has wide-spread expression throughout our tissues. Its thioredoxin motif suggests oxidoreductase function; however, its mechanism and functional role(s) are still being uncovered. Furthermore, the connection of both high and low expression levels of SELENOM to separate diseases emphasizes the medical application for studying the role of Sec in this protein. In this review, we aim to decipher the role of SELENOM through detailing and connecting current evidence. With multiple proposed functions in diverse tissues, continued research is still necessary to fully unveil the role of SELENOM.


Neurobiological Markers for Predicting Treatment Response in Patients with Bipolar Disorder.
Predictive neurobiological markers for prognosis are essential but underemphasized for patients with bipolar disorder (BD), a neuroprogressive disorder. Hence, we developed models for predicting symptom and functioning changes. Sixty-one patients with BD were recruited and assessed using the Young Mania Rating Scale (YMRS), Montgomery−Åsberg Depression Rating Scale (MADRS), Positive and Negative Syndrome Scale (PANSS), UKU Side Effect Rating Scale (UKU), Personal and Social Performance Scale (PSP), and Global Assessment of Functioning scale both at baseline and after 1-year follow-up. The models for predicting the changes in symptom and functioning scores were trained using data on the brain morphology, functional connectivity, and cytokines collected at baseline. The correlation between the predicted and actual changes in the YMRS, MADRS, PANSS, and UKU scores was higher than 0.86 (q < 0.05). Connections from subcortical and cerebellar regions were considered for predicting the changes in the YMRS, MADRS, and UKU scores. Moreover, connections of the motor network were considered for predicting the changes in the YMRS and MADRS scores. The neurobiological markers for predicting treatment-response symptoms and functioning changes were consistent with the neuropathology of BD and with the differences found between treatment responders and nonresponders.


The influence of the final cause doctrine on anatomists of the sixteenth and seventeenth centuries concerning selected anatomical structures of the head and neck.
The Doctrine of Final Cause, taken from Aristotle's "causes" and modified by Claudius (Aelius) Galen (of Pergamon) stated that for an anatomical part to exist it must have a "cause," not an end point, but a purpose or goal, natural or divine. This affected the renaissance anatomist's thinking. We explore this doctrine's relationship with human head and neck anatomy from antiquity's Aristotle and Galen, and the leading renaissance anatomists from the 16th and 17th centuries. Their relevant writings were influenced by religious and political beliefs and varied from humanistic to reactionary. Tracing anatomical controversies through these works reveal the humanism of Vesalius and others as paralleling the humanists of art and literature. These controversies illustrate how the body was used to demonstrate function, uses, and causes from higher sources. Humanists advanced the social, philosophical, intellectual, literary, and medical/anatomical thoughts of this period. They stood between the Christian church of the Middle Ages and modern science. Like religion, medicine and anatomy had its own revealed sources of knowledge and had sacred texts like Galen's. Vesalius' the Fabrica and the woodcuts established suddenly the beginning of modern observational science and art as the direct and faithful representation of natural phenomena. They displayed anatomy such that others could understand, including errors of Galen, bringing Vesalius into ecclesiastical conflict. Evolutionary scientists today see mutations as favorable or unfavorable depending on the environment. Mutations are random or directed by divine plan, according to perspectives of this ancient debate.


Follicle Microstructure and Innervation Vary between Pinniped Micro- and Macrovibrissae.
Histological data from terrestrial, semiaquatic, and fully aquatic mammal vibrissa (whisker) studies indicate that follicle microstructure and innervation vary across the mystacial vibrissal array (i.e. medial microvibrissae to lateral macrovibrissae). However, comparative data are lacking, and current histological studies on pinniped vibrissae only focus on the largest ventrolateral vibrissae. Consequently, we investigated the microstructure, medial-to-lateral innervation, and morphometric trends in harp seal (Pagophilus groenlandicus) vibrissal follicle-sinus complexes (F-SCs). The F-SCs were sectioned either longitudinally or in cross-section and stained with a modified Masson's trichrome stain (microstructure) or Bodian's silver stain (innervation). All F-SCs exhibited a tripartite blood organization system. The dermal capsule thickness, the distribution of major branches of the deep vibrissal nerve, and the hair shaft design were more symmetrical in medial F-SCs, but these features became more asymmetrical as the F-SCs became more lateral. Overall, the mean axon count was 1,221 ± 422.3 axons/F-SC and mean axon counts by column ranged from 550 ± 97.4 axons/F-SC (medially, column 11) to 1,632 ± 173.2 axons/F-SC (laterally, column 2). These values indicate a total of 117,216 axons innervating the entire mystacial vibrissal array. The mean axon count of lateral F-SCs was 1,533 ± 192.9 axons/ F-SC, which is similar to values reported in the literature for other pinniped F-SCs. Our data suggest that conventional studies that only examine the largest ventrolateral vibrissae may overestimate the total innervation by ∼20%. However, our study also accounts for variation in quantification methods and shows that conventional analyses likely only overestimate innervation by ∼10%. The relationship between axon count and cross-sectional F-SC surface area was nonlinear, and axon densities were consistent across the snout. Our data indicate that harp seals exhibit microstructural and innervational differences between their microvibrissae (columns 8-11) and macrovibrissae (columns 1-7). We hypothesize that this feature is conserved among pinnipeds and may result in functional compartmentalization within their mystacial vibrissal arrays.


Referral of patients to an anticoagulant clinic: implications for better management.
The quality of anticoagulant treatment of ambulatory patients is affected by the content of referral letters and administrative processes. To assess these influences a method was developed to audit against the hospital standard the referral of patients to one hospital anticoagulant clinic in a prospective study of all (80) new patients referred to the clinic over eight months. Administrative information was provided by the clinic coordinator, and the referral letters were audited by the researchers. Referral letters were not received by the clinic for 10% (8/80) of patients. Among the 72 referral letters received, indication for anticoagulation and anticipated duration of treatment were specified in most (99%, 71 and 81%, 58 respectively), but only 3% (two) to 46% (33) reported other important clinical information (objective investigations, date of starting anticoagulation, current anticoagulant dose, date and result of latest international normalised ratio, whether it should be the anticoagulant clinic that was eventually to stop anticoagulation, patients' other medical problems and concurrent treatment. Twenty two per cent (16/80) of new attenders were unexpected at the anticoagulant clinic. Most patients' case notes were obtained for the appointment (61%, 47/77 beforehand and 30% 23/77 on the day), but case notes were not obtained for 9% (7/77). The authors conclude that health professionals should better appreciate the administrative and organisational influences that affect team work and quality of care. Compliance with a well documented protocol remained below the acceptable standard. The quality of the referral process may be improved by using a more comprehensive and helpful referral form, which has been drawn up, and by educating referring doctors. Measures to increase the efficiency of the administrative process include telephoning the clinic coordinator directly, direct referrals through a computerised referral system, and telephone reminders by haematology office staff to ward staff to ensure availability of the hospital notes. The effect of these changes will be assessed in a repeat audit.


Optimized removal of the tongue-and-groove underdose via constrained partial synchronization and variable depth recursion.
Full synchronization (FS) leaf sequencing removes tongue-and-groove underdosages (TGU) but increases the number of segments. Constrained partial synchronization (CPS) uses a minimum tongue-and-groove ratio (TGR) to reduce the number of segments while achieving acceptable TGUs. TGR is the ratio of non-overlapping intensities that irradiate a common junction. For TGRs of 1, 1.5 and 2, the TGUs were measured as 18%, 4% and 0%, respectively, for a 6 MV beam and a Siemens 82 leaf MLC. The extraction and sweep processes of the variable depth recursion (VDR) leaf-sequencing algorithm were constrained to satisfy a minimum TGR. For a Siemens MLC and 15 clinical cases, VDR with a TGR = 1.5 produced 7% more segments than the unsynchronized VDR, while a fully synchronized sweeping window algorithm produces 62% more segments. For random intensity maps, VDR with CPS produced significantly fewer segments than an unsynchronized sweeping window. Similar results can be obtained for MLCs that interdigitate. This has implications for direct aperture optimization algorithms (DAO) that use the sweeping window as a starting point (Pinnacle), for which a significant TGU has been observed. The concept of CPS can be applied to DAO by choosing appropriate levels for each of the segments in DAO.


Failure to demonstrate high concentrations of luteinising hormone-releasing hormone in the bovine pineal body.
Six in vitro experiments were conducted to study the effects of the incorporation into the culture system of 10 bovine pineal bodies (or a similar mass of bovine hypothalamic tissue) on the secretion of luteinising hormone (LH) from bovine adenohypophyseal tissue (anterior pituitary - AP). When hypothalamic tissue was incubated with AP tissue, an increased amount of LH was released into the medium but no marked increase in LH secretion was observed when pineal bodies were included in the AP culture system. Additionally, the amount of radio-immunologically active luteinising hormone-releasing hormone (LHRH) contained in bovine pineal bodies was much less than that contained in an equivalent mass of hypothalamic tissue. Thus, present results do not support the concept that the pineal body contains comparatively large amounts of lhrh.


Electrophysiological Investigation of Different Methods of Anesthesia in Lobster and Crayfish.
In search for methods of anesthesia of crustaceans, an implanted electrode into lobster and crayfish CNS enabled us to monitor signal propagation in the nerve system of animals undergoing different protocols. Cooling (tap water 0°C, sea water -1,8°C) and anesthesia with MgCl2 (10%) were both discarded as anesthetic procedures because responses to external stimuli were still detectable under treatment. Contrarily, bubbling the aquarium water with CO2 can be considered a "partially successful" anesthesia, because signal propagation is inhibited but before that the animals show discomfort. The procedure of "electro-stunning" induces epileptic-form seizures in the crustacean CNS (lobster, crayfish), which overlay but do not mitigate the response to external stimuli. After several minutes the activity declines before the nervous system starts to recover. A feasible way to sacrifice lobsters is to slowly raise the water temperature (1°C min-1), as all electrical activities in the CNS cease at temperatures above ~30°C, whereas below this temperature the animals do not show signs of stress or escape behavior (e.g. tail flips) in the warming water. CO2 is efficient to anaesthetize lobster and crayfish but due to low pH in water is stressful to the animals previous to anesthesia. Electrical stunning induces epileptiform seizures but paralyses the animals and leads to a reversible decline of nerve system activity after seizure. Electric stunning or slowly warming just before preparation may meet ethical expectations regarding anaesthesia and to sacrifice crustaceans.


Influence of N1-Methylpseudouridine in Guide RNAs on CRISPR/Cas9 Activity.
At present, there are many strategies to improve the activity of CRISPR/Cas9. A well-known and effective approach is guide RNA modification. Many chemical guide RNA modifications have been studied, whereas naturally occurring RNA modifications are largely unexplored. N1-methylpseudouridine (m1Ψ) is an RNA base modification widely used in mRNA therapy, and it holds great promise for application in genome editing systems. The present study focuses on investigating the effect of N1-methylpseudouridine on the functioning of CRISPR/Cas9. In vitro cleavage assays helped determine the level of m1Ψ guide RNA modification that is sufficient to cleave the target substrate. By analyzing FAM-labeled dsDNA substrate cleavage, we calculated the kinetic parameters and the specificity scores of modified guide RNAs. Neon transfection and digital PCR enabled us to assess the activity of modified guide RNAs in mammalian cells. Our study shows that the presence of m1Ψ in guide RNAs can help preserve on-target genome editing while significantly reducing the off-target effects of CRISPR/Cas9 in vitro. We also demonstrate that Cas9 complexes with guide RNAs containing m1Ψ allow for genome editing in human cells. Thus, the incorporation of m1Ψ into guide RNAs supports CRISPR/Cas9 activity both in vitro and in cells.


Strength and size of phosphorus-rich patches determine the foraging strategy of Neyraudia reynaudiana.
Under natural conditions, soil nutrients are heterogeneously distributed, and plants have developed adaptation strategies to efficiently forage patchily distributed nutrient. Most previous studies examined either patch strength or patch size separately and focused mainly on root morphological plasticity (increased root proliferation in nutrient-rich patch), thus the effects of both patch strength and size on morphological and physiological plasticity are not well understood. In this study, we examined the foraging strategy of Neyraudia reynaudiana (Kunth) Keng ex Hithc, a pioneer grass colonizing degraded sites, with respect to patch strength and size in heterogeneously distributed phosphorus (P), and how foraging patchily distributed P affects total plant biomass production. Plants were grown in sand-culture pots divided into ½, ¼, 1/6 compartments and full size and supplied with 0 + 0/30, 0 + 7.5/30 and 7.5 + 0/30 mg P/kg dry soil as KH2PO4 or 0 + 15/15, 0 + 18.5/ 18.5, 7.5 + 15/15 mg kg - 1 in the homogenous treatment. The first amount was the P concentration in the central region, and that the second amount was the P concentration in the outer parts of the pot. After 3 months of growth under experimental conditions, significantly (p < 0.05) high root elongation, root surface area, root volume and average root diameter was observed in large patches with high patch strength. Roots absorbed significantly more P in P-replete than P-deficient patches. Whole plant biomass production was significantly higher in larger patches with high patch strength than small patches and homogeneous P distribution. The result demonstrates that root morphological and physiological plasticity are important adaptive strategies for foraging patchily distributed P and the former is largely determined by patch strength and size. The results also establish that foraging patchily distributed P resulted in increased total plant biomass production compared to homogeneous P distribution.


Neuropsychological evidence for abnormal neurodevelopment associated with early-onset psychoses.
The longitudinal neuropsychological study of first-episode early-onset psychosis (EOP) patients, whose brain maturation is still in progress at the time of illness onset, provides a unique opportunity to compare their cognitive development with that of healthy subjects, in search of specific patterns resulting from the interaction between neurodevelopmental processes and the presence of psychotic disorders. Method Seventy-five first-episode EOP patients (schizophrenia n = 35; bipolar disorder n = 17; other forms of psychosis n = 23) with a mean age of 15.53 years were assessed with a neuropsychological battery that included measures of attention, working memory, memory and executive functions within 6 months following the onset of the first psychotic symptom (baseline) and 2 years later. Psychotic symptoms were assessed at both times with the Positive and Negative Symptom Scale (PANSS). Seventy-nine healthy subjects matched for age and education served as controls. EOP patients showed significant cognitive impairment at both baseline and the 2-year follow-up, with no significant differences between diagnostic groups at either time. Both healthy controls and EOP patients improved in all cognitive measures, except for patient working memory. Improvement in patient attention lost significance after controlling for psychotic symptom reduction. No significant time/diagnosis interaction was found among patients (p > 0.405). Cognitive impairment in EOP is already present at the first episode, and cognitive development seems to be arrested early in EOP patients compared to their healthy peers, at least for some cognitive functions. These and previous similar results support the neurodevelopmental hypothesis of psychosis.


Immediate Dental Implant Placement in the Oncologic Setting: A Conceptual Framework.
Historically, immediate dental implants have been reserved for patients with benign disease, with full dental rehabilitation rarely being accomplished in the oncologic setting due to concerns related to implant survival, flap compromise, and delay in initiation of adjuvant therapy. Recent developments in technology have made immediate dental implants using virtual surgical planning safe and reliable. At Memorial Sloan Kettering Cancer Center, we have implemented a workflow for immediate dental implant placement in the oncologic patient population that has become a routine part of maxillary and mandibular reconstruction. This approach begins with a multidisciplinary virtual surgical planning session and custom dental splints to be used for cutting and inset guides. Dental implants are placed intraoperatively at the time of tumor resection and reconstruction with the fibula flap. A temporary prosthesis, which can be worn during radiation therapy, is placed following a vestibuloplasty, approximately 4-6 weeks after the initial reconstruction. After the completion of radiation therapy and the resolution of edema, a permanent prosthesis is placed. When critically evaluating our experience, we have found that patients undergoing immediate dental implant placement have higher rates of implant survival and no delay in adjuvant therapy. The protocol described here in detail has successfully expanded the indications for immediate dental rehabilitation in the oncologic patient population.


Evaluation of two treatment concepts of four implants supporting fixed prosthesis in an atrophic maxilla: finite element analysis.
Currently, oblique placement of long implants or the use of short implants to circumvent the maxillary sinus area and provide support for fixed prostheses are viable alternatives. The purpose of this study was to compare these two treatment concepts and ascertain which one exhibits superior biomechanical characteristics. Two different treatment concept models were constructed. The first one, LT4I, consisting of two mesial vertical implants positioned in lateral incisor regions and two distal tilted implants (45°) situated in second premolar regions of the maxilla. The second model, VS4I, includes two mesial vertical implants in lateral incisor regions and two vertically positioned short implants in second premolar regions. Numerical simulations were conducted under three loading types: firstly, oblique forces upon the molars; secondly, vertical forces upon the molars; thirdly, oblique forces upon the incisors. The maximum principal stress (σmax) and minimum principal stress (σmin) of the bone, as well as von Mises stress of the implants, were calcuated. Under oblique loading on the molar, higher stress values in the bone were observed in LT4I group. Under vertical loading on molar, higher stress values in the bone were also observed in LT4I group. Furthermore, little difference was found between the two groups under oblique loading on the incisor. Both treatment concepts can be applicable for edentulous individuals with moderate atrophic maxilla. Compared to tilted implants, short implants can transmit less occlusal force to the supporting tissues.


Activity of lenalidomide in mantle cell lymphoma can be explained by NK cell-mediated cytotoxicity.
Lenalidomide is an immunomodulatory agent that has demonstrated clinical benefit for patients with relapsed or refractory mantle cell lymphoma (MCL); however, despite this observed clinical activity, the mechanism of action (MOA) of lenalidomide has not been characterized in this setting. We investigated the MOA of lenalidomide in clinical samples from patients enrolled in the CC-5013-MCL-002 trial (NCT00875667) comparing single-agent lenalidomide versus investigator's choice single-agent therapy and validated our findings in pre-clinical models of MCL. Our results revealed a significant increase in natural killer (NK) cells relative to total lymphocytes in lenalidomide responders compared to non-responders that was associated with a trend towards prolonged progression-free survival and overall survival. Clinical response to lenalidomide was independent of baseline tumour microenvironment expression of its molecular target, cereblon, as well as genetic mutations reported to impact clinical response to the Bruton tyrosine kinase inhibitor ibrutinib. Preclinical experiments revealed lenalidomide enhanced NK cell-mediated cytotoxicity against MCL cells via increased lytic immunological synapse formation and secretion of granzyme B. In contrast, lenalidomide exhibited minimal direct cytotoxic effects against MCL cells. Taken together, these data provide the first insight into the clinical activity of lenalidomide against MCL, revealing a predominately immune-mediated MOA.


Tolterodine extended release improves overactive bladder symptoms in men with overactive bladder and nocturia.
To evaluate the efficacy and safety of nighttime dosing with tolterodine extended release (TER) in men with overactive bladder (OAB) and nocturia. This was a post hoc analysis of data from two 12-week, double-blind, placebo-controlled trials of nighttime (<4 hours before bedtime) TER (4 mg daily) dosing. Men with a mean micturition frequency of eight or more times in 24 hours, including a mean of 2.5 or more nocturia episodes/night, were included. For each micturition, patients used 7-day diaries to record urinary urgency on a 5-point urgency rating scale (1, none; 2, mild; 3, moderate; 4, severe; 5, urgency urinary incontinence). Micturitions were analyzed post hoc by urgency rating categories: total (1 to 5), non-OAB (1 to 2), OAB (3 to 5), and severe OAB (4 to 5). Adverse events were recorded throughout the study. A total of 745 men (mean age 64 years) were randomized to placebo (n = 374) or TER (n = 371). Of the 745 men, 73% reported no incontinence episodes in a 7-day diary at baseline. At week 12, the weekly values for nighttime severe OAB micturitions and 24-hour and daytime total, OAB, and severe OAB micturitions were significantly reduced in the TER group versus the placebo group. The TER-treated men also reported a significant reduction in the mean urgency rating versus placebo. Adverse events associated with TER were low and comparable to those in the placebo group, with the exception of dry mouth (11% versus 4%). Withdrawals because of adverse events were infrequent (3% TER, 4% placebo). Five men were withdrawn for symptoms suggestive of urinary retention (3 TER, 2 placebo). Nighttime TER dosing reduced urgency-related micturitions and was well tolerated in men with OAB and nocturia.


The lipid-mobilizing effect of atrial natriuretic peptide is unrelated to sympathetic nervous system activation or obesity in young men.
We recently demonstrated that natriuretic peptides and especially the atrial natriuretic peptide (ANP) are powerful lipolytic agents on isolated human fat cells. To search for a possible influence of obesity on ANP responsiveness, we compared the lipolytic effects of human ANP (h-ANP) on isolated subcutaneous abdominal adipose tissue (SCAAT) fat cells from young healthy lean and obese men. The lipid-mobilizing effects of an intravenous infusion of h-ANP was studied, as well as various metabolic and cardiovascular parameters that were compared in the same subjects. h-ANP (50 ng/min/kg) was infused iv for 60 min. Microdialysis probes were inserted in SCAAT to measure modifications of the extracellular glycerol concentrations during h-ANP infusion. Spectral analysis of blood pressure and heart rate oscillations that were recorded using digital photoplethysmography were used to assess changes in autonomic nervous system activity. h-ANP induced a marked and similar increase in glycerol and nonesterified fatty acids, and a weak increase in insulin plasma levels in lean and obese men. Plasma norepinephrine concentrations rose similarly during h-ANP infusion in lean and obese men. The effects of h-ANP infusion on the autonomic nervous system were similar in both groups, with an increase in the spectral energy of the low-frequency band of systolic blood pressure variability and a decrease in the spectral energy of the high-frequency band of heart rate. In SCAAT, h-ANP infusion increased extracellular glycerol concentration and decreased blood flow similarly in both groups. The increase in extracellular glycerol observed during h-ANP infusion was not modified when 0.1 mM propranolol was added to the microdialysis probe perfusate to prevent beta-adrenoceptor activation. These data show that ANP is a potent lipolytic hormone independent of the activation of the sympathetic nervous system, and that obesity did not modify the lipid-mobilizing effect of ANP in young obese subjects.


Evaluation of satisfaction and complications in patients with existing complete dentures.
The aim of this clinical trial was to assess satisfaction, as well as the frequency and type of prosthetic complications in terms of several variables, in patients with complete dentures that had been supplied at private clinics. The study subjects were 64 patients with a mean age of 63.48 years wearing complete dentures provided at private clinics, and requesting new ones. The degree of patient satisfaction with their dentures was assessed, as well as complications and parameters related to the dentures such as the accuracy of vertical dimensions and centric relation, arrangement and possible malposition of the artificial teeth, and the border length of the denture bases. The most common complication was loss of retention (85.9%), followed by ulceration (44.2%). Mandibular dentures with long vestibular borders showed a significantly higher incidence of epulis fissuratum (P = 0.017), and denture-related sore spots influenced patients' speech ability (P = 0.023). Routine recalls seem to be important for wearers of complete dentures, as several insidious complications may develop and cause damage to the dentures as well as the patients' oral tissues.


Maternal immune activation and strain specific interactions in the development of autism-like behaviors in mice.
It is becoming increasingly apparent that the causes of autism spectrum disorders (ASD) are due to both genetic and environmental factors. Animal studies provide important translational models for elucidating specific genetic or environmental factors that contribute to ASD-related behavioral deficits. For example, mouse research has demonstrated a link between maternal immune activation and the expression of ASD-like behaviors. Although these studies have provided insights into the potential causes of ASD, they are limited in their ability to model the important interactions between genetic variability and environmental insults. This is of particular concern given the broad spectrum of severity observed in the human population, suggesting that subpopulations may be more susceptible to the adverse effects of particular environmental insults. It is hypothesized that the severity of effects of maternal immune activation on ASD-like phenotypes is influenced by the genetic background in mice. To test this, pregnant dams of two inbred strains (that is, C57BL/6J and BTBR T(+)tf/J) were exposed to the viral mimic polyinosinic-polycytidylic acid (polyI:C), and their offspring were tested for the presence and severity of ASD-like behaviors. To identify differences in immune system regulation, spleens were processed and measured for alterations in induced cytokine responses. Strain-treatment interactions were observed in social approach, ultrasonic vocalization, repetitive grooming and marble burying behaviors. Interestingly, persistent dysregulation of adaptive immune system function was only observed in BTBR mice. Data suggest that behavioral and immunological effects of maternal immune activation are strain-dependent in mice.


Lymph nodes in gastric cancer.
Surgery is the only curative therapy for gastric cancer and controversy still exist on the extend of surgery. As the lymphatic distribution of stomach is very complex, the determination of the actual lymph node involvement is important for making the decision in order to avoid complications. Sentinel node navigation surgery has recently been introduced in gastrointestinal tract cancer. Present article reviews the detection techniques of lymph nodes and significance of lymphadenectomies in gastric cancer.


Mitochondrial targeting dendrimer allows efficient and safe gene delivery.
Cationic dendrimers are promising tools in the introduction of nucleic acids into mammalian cells, however, present dendrimer-based gene vectors have modest transfection efficacy and significant toxicity. Here, we synthesized triphenylphosphonium (TPP) conjugated dendrimers and used them as polymeric gene vectors. TPP-conjugated dendrimers can effectively target mitochondria and showed efficient transfection efficacy on HeLa and COS-7 cells with low cytotoxicity on the transfected cells. These materials achieved much higher efficacy than the commercial transfection reagent SuperFect and comparable efficacy with Lipofectamine 2000. This study provides a new insight into the design of efficient and biocompatible gene vectors using dendrimers as scaffolds.


Discrete choice experiment to evaluate preferences of patients with cystic fibrosis among alternative treatment-related health outcomes: a protocol.
Clinical decision-making is a complex process. Patient preference information regarding desirable health states should inform treatment and is critical to agreeing on goals of therapy. Cystic fibrosis (CF) is a common, inheritable multisystem disorder for which the major manifestation is progressive, chronic lung disease. Intermittent pulmonary exacerbations are a hallmark of disease and these drive lung damage that results in premature death. We suspect that clinicians make assumptions, most likely implicit assumptions, about outcomes that are desired by patients who are treated for pulmonary exacerbations. The aim of this study is to identify and quantify the preferences of patients with cystic fibrosis regarding treatment outcomes. We will develop a discrete choice experiment (DCE) in collaboration with people with CF and their carers, and evaluate how patients make trade-offs between different aspects of health-related status when considering treatment options. Ethics approval for all aspects of this study was granted by the Western Australia Child and Adolescent Health Service Human Research Ethics Committee [RGS903]. Weighted preference information from the DCE will be used to develop a multiattribute utility instrument as a measure of treatment success in the upcoming Bayesian Evidence-Adaptive Trial to optimise management of CF. Dissemination of results will also occur through peer-reviewed publications and presentations to relevant stakeholders and research networks.


[Analysis of genetic penetrance in the pedigree of cases with Van der Woude syndrome: report of five cases].
Van der Woude syndrome is a congenital abnormality characterized by labial cysts, accessory salivary glands, pits, fistulas and paramedian sinuses of the lower lips, and is frequently associated with cleft lip and palate. This disease is the most common syndromic cleft abnormality. The disease is characterized by a single gene abnormality where craniofacial morphogenesis is affected. It has an autosomal dominant inheritance with involvement of 1q32-41 chromosomal locus. However, the gene expression profile is variable and the disease may present in some individuals with sinuses of the lower lips only. We present results of analysis of genetic penetrance in the pedigree of five cases with Van der Woude syndrome together with a review of the literature.


A Gas-Powered, Patient-Responsive Automatic Resuscitator for Use in Acute Respiratory Failure: A Bench and Experimental Study.
During the COVID-19 pandemic, a need for innovative, inexpensive, and simple ventilator devices for mass use has emerged. The Oxylator (CPR Medical Devices, Markham, Ontario, Canada) is an FDA-approved, fist-size, portable ventilation device developed for out-of-hospital emergency ventilation. It has not been tested in conditions of severe lung injury or with added PEEP. We aimed to assess the performance and reliability of the device in simulated and experimental conditions of severe lung injury, and to derive monitoring methods to allow the delivery of safe, individualized ventilation during situations of surge. We bench-tested the functioning of the device with an added PEEP valve extensively, mimicking adult patients with various respiratory mechanics during controlled ventilation, spontaneous breathing, and prolonged unstable conditions where mechanics or breathing effort was changed at every breath. The device was further tested on a porcine model (4 animals) after inducing lung injury, and these results were compared with conventional ventilation modes. The device was stable and predictable, delivering a constant flow (30 L/min) and cycling automatically at the inspiratory pressure set (minimum of 20 cm H2O) above auto-PEEP. Changes in respiratory mechanics manifested as changes in respiratory timing, allowing prediction of tidal volumes from breathing frequency. Simulating lung injury resulted in relatively low tidal volumes (330 mL with compliance of 20 mL/cm H2O). In the porcine model, arterial oxygenation, CO2, and pH were comparable to conventional modes of ventilation. The Oxylator is a simple device that delivered stable ventilation with tidal volumes within a clinically acceptable range in bench and porcine lung models with low compliance. External monitoring of respiratory timing is advisable, allowing tidal volume estimation and recognition of changes in respiratory mechanics. The device can be an efficient, low-cost, and practical rescue solution for providing short-term ventilatory support as a temporary bridge, but it requires a caregiver at the bedside.


Experimental validation of a modeling framework for upconversion enhancement in 1D-photonic crystals.
Photonic structures can be designed to tailor luminescence properties of materials, which becomes particularly interesting for non-linear phenomena, such as photon upconversion. However, there is no adequate theoretical framework to optimize photonic structure designs for upconversion enhancement. Here, we present a comprehensive theoretical model describing photonic effects on upconversion and confirm the model's predictions by experimental realization of 1D-photonic upconverter devices with large statistics and parameter scans. The measured upconversion photoluminescence enhancement reaches 82 ± 24% of the simulated enhancement, in the mean of 2480 separate measurements, scanning the irradiance and the excitation wavelength on 40 different sample designs. Additionally, the trends expected from the modeled interaction of photonic energy density enhancement, local density of optical states and internal upconversion dynamics, are clearly validated in all experimentally performed parameter scans. Our simulation tool now opens the possibility of precisely designing photonic structure designs for various upconverting materials and applications.


Complex self-management interventions in chronic disease unravelled: a review of lessons learned from an individual patient data meta-analysis.
Meta-analyses using individual patient data (IPD) rather than aggregated data are increasingly applied to analyze sources of heterogeneity between trials and have only recently been applied to unravel multicomponent, complex interventions. This study reflects on methodological challenges encountered in two IPD meta-analyses on self-management interventions in patients with heart failure or chronic obstructive pulmonary disease. Critical reflection on prior IPD meta-analyses and discussion of literature. Experience from two IPD meta-analyses illustrates methodological challenges. Despite close collaboration with principal investigators, assessing the effect of characteristics of complex interventions on the outcomes of trials is compromised by lack of sufficient details on intervention characteristics and limited data on fidelity and adherence. Furthermore, trials collected baseline variables in a highly diverse way, limiting the possibilities to study subgroups of patients in a consistent manner. Possible solutions are proposed based on lessons learnt from the methodological challenges. Future researchers of complex interventions should pay considerable attention to the causal mechanism underlying the intervention and conducting process evaluations. Future researchers on IPD meta-analyses of complex interventions should carefully consider their own causal assumptions and availability of required data in eligible trials before undertaking such resource-intensive IPD meta-analysis.


ACR Appropriateness Criteria: local excision in early-stage rectal cancer.
Low anterior resection or abdominoperineal resection is considered standard treatment for early rectal cancer. These procedures, however, carry a risk of morbidity and mortality that may not be warranted for early distal lesions, which may be treated with local excision. Emerging data has investigated the efficacy of local excision in patients with early stage rectal cancers. An expert panel designated by the American College of Radiology has reviewed supporting data, from a few prospective multi-institutional trials and a number of single-institution, retrospective reviews. The consensus recognizes the importance of accurate staging to identify patients who may be candidates for a local excision approach. Optimal candidates for local excision alone include small, low-lying T1 tumors, without adverse pathologic features. A number of procedures may be safely used including transanal, posterior trans-sphincteric, posterior proctotomy, transanal excision, or transanal microsurgery. It is important to note that none of these include lymph node evaluation, and depending on the risk of lymph node metastases, adjuvant radiation with or without chemotherapy may be warranted. Patients with positive margins or T3 lesions are at high risk of local recurrence and should be offered immediate APR or LAR. However, patients with high-risk T1 tumors, T2 tumors, or those who are not amenable to more radical surgery may benefit from adjuvant treatment. Data have also reported excellent local control rates for neoadjuvant radiation +/- chemotherapy followed by local excision in higher risk patients, but it is not yet clear if this approach reduces recurrence rates over surgery alone.


Peptide growth factors and their receptors in the vein wall.
The varicose vein wall remodeling is a very complex process, which is controlled by numerous factors, including peptide growth factors. The aim of the study was to assess a/b FGF, IGF-1, TGF-β1, VEGF-A and their receptors in the vein wall. Varicose vein samples were taken from 24 patients undergoing varicose vein surgery. The control material consisted of vein specimens collected from 12 patients with chronic limb ischemia. Contents of aFGF, bFGF, IGF-I, TGF-β1, VEGF, IGF-1R, VEGF R1 and VEGF R2 were assessed with ELISA method. Protein expression of FGF R1 and TGF-β RII were evaluated with western blot. Increased contents of aFGF, IGF-1 and VEGF-A were found in varicose veins in comparison with normal ones (p<0.05). In contrast, a significant decrease in TGF-β content was demonstrated in varicose veins (p<0.05). Furthermore, there was no difference in bFGF content in both groups (p>0.05). IGF-1 R content was significantly increased in varicose veins (p<0.05). There was no difference in VEGF R1 content between varicose and normal veins (p>0.05), whereas VEGF R2 content was significantly increased in varicose veins (p<0.05). Western blot demonstrated increased expression of TGF-β RII in varicose veins (p<0.05) and similar expression of FGF R1 in both groups (p>0.05). Demonstrated changes in peptide growth factors and their receptors may disturb metabolism of extracellular matrix in the varicose vein wall and contribute to the development of the disease to its more advanced stages.


Accuracy of a point-of-care test for quantifying human chorionic gonadotrophin (hCG) in the management of pregnancy of unknown location in an abortion service.
Women may seek abortion at gestations when there is no visible intrauterine pregnancy on ultrasound scanning. Clinical protocols for pregnancy of unknown location (PUL) require measurement of serum human chorionic gonadotrophin (hCG), with the National Institute for Health and Care Excellence recommending that values above 1500 IU/L be further investigated to exclude ectopic pregnancy. Our aim was to determine whether a point-of-care test (POCT) could be used instead of laboratory serum hCG measurement. Over 12 months, women who presented to an abortion service with a PUL had a POCT for blood or urine hCG and laboratory serum hCG measurement. The POCT machine used provides a discrete hCG value below 1000 IU/L and above this gives results as a range. The sensitivity and specificity of the POCT in identifying cases where laboratory serum hCG results were above 1500 IU/L were calculated. A total of 118 women presented with a PUL, of whom 70 had a POCT on blood (n=49) or urine (n=21) and a corresponding laboratory serum hCG. The sensitivity of the blood POCT was 0.67 (95% CI 0.38 to 0.87) and the specificity was 0.97 (95% CI 0.83 to 0.99). The sensitivity of the urine POCT was 0.25 (CI 0.01 to 0.78) and the specificity was 0.94 (CI 0.69 to 0.99). Although both the blood and urine POCTs had a high level of specificity, neither test was acceptably sensitive. While promising, this POCT for hCG is not sufficiently reliable to replace laboratory serum hCG testing in the management of women with PUL in an abortion service.


LRRK2 mutant iPSC-derived DA neurons demonstrate increased susceptibility to oxidative stress.
Studies of Parkinson's disease (PD) have been hindered by lack of access to affected human dopaminergic (DA) neurons. Here, we report generation of induced pluripotent stem cells that carry the p.G2019S mutation (G2019S-iPSCs) in the Leucine-Rich Repeat Kinase-2 (LRRK2) gene, the most common PD-related mutation, and their differentiation into DA neurons. The high penetrance of the LRRK2 mutation and its clinical resemblance to sporadic PD suggest that these cells could provide a valuable platform for disease analysis and drug development. We found that DA neurons derived from G2019S-iPSCs showed increased expression of key oxidative stress-response genes and α-synuclein protein. The mutant neurons were also more sensitive to caspase-3 activation and cell death caused by exposure to stress agents, such as hydrogen peroxide, MG-132, and 6-hydroxydopamine, than control DA neurons. This enhanced stress sensitivity is consistent with existing understanding of early PD phenotypes and represents a potential therapeutic target.


Anaerobic dormancy quantified in artemia embryos: a calorimetric test of the control mechanism.
Continuous measurement of heat dissipation from brine shrimp embryos during reversible transitions from aerobic development to anaerobic dormancy demonstrates a primary role for intracellular pH(pH(i))in this metabolic switching. Artificially elevating the depressed pH(i) during anoxia by adding ammonia markedly reactivates metabolism, as judged by increases in heat dissipation, trehalose catabolism, and the ratio of adenosine triphosphate to adenosine diphosphate. Energy flow during anaerobic dormancy is suppressed to 2.4 percent of aerobic values, which is the lowest percentage thus far reported for euryoxic animals. Use of diguanosine tetraphosphate stores cannot account for this observed heat dissipation. Thus, mobilizing trace amounts of trehalose may explain the energy metabolism during quiescence.


Risk factors and prognostic implications for pathologic upstaging to T3a after partial nephrectomy.
Performing partial nephrectomy (PN) on a cT1 tumor, which postoperatively is upgraded to pT3a can possibly lead to compromise of cancer specific mortality. We therefore aimed to identify risk factors for pathologic T3a upstaging of cT1 tumors and to analyze the association between upstaging, positive surgical margins (PSM) and overall survival (OS). The present study included patients who underwent PN for a clinically localized T1 renal mass from two datasets: 1) 1298 patients from a prospectively maintained multi-center database (MCDB); and 2) 7940 patients from the National Cancer Database (NCDB). Multivariable logistic regression models within each cohort were used to identify predictors of cT1 to pT3a upstaging and its association with PSM. Cox proportion hazards regression models were used to compare overall survival in the NCDB cohort. The rate of pT3a upstaging was 5.7% (N.=74) in the MCDB and 1.9% (N.=156) in the NCDB cohort. Older age (MCDB OR=1.04, P=0.001; NCDB OR=1.04, P=0.001) and larger tumor size (MCDB OR=1.89, P<0.001; NCDB OR=1.38, P<0.001) increased the likelihood of upstaging. PSM was found to be more likely for pT3a upstaged patients in both cohorts (MCDB 14.9% vs. 3.5%, P<0.001; NCDB 14.8% vs. 8.3%, P=0.006), even when adjusting for tumor size. At short term follow-up (NCDB median follow-up 27.3 months), pT3a upstaging was associated with worse OS in univariable (HR=1.89; 95% CI=1.00, 3.55; P=0.049) but not multivariable analysis (HR=1.63; 95% CI=0.86, 3.08; P=0.131). OS was 93.0% vs. 95.8% at 3 years for those with and without pT3a upstaging, respectively. Larger tumor size and increased age are associated with pathological upstaging to T3a for clinical T1 tumors treated with partial nephrectomy. Steps to improve identification of occult pT3a disease are necessary as its occurrence significantly increased the likelihood of a PSM, both in a high-volume multicenter cohort, as well as, a national data registry.


Intermittent Hypoxia Does not Elicit Memory Impairment in Spinal Cord Injury Patients.
There is a critical need for new therapeutic strategies to restore motor function in patients with spinal cord injuries (SCIs), without unwanted effects. Intermittent hypoxia (IH) induces plasticity in spared synaptic pathways to motor neurons below the level of injury, which can be harnessed to elicit motor recovery in incomplete SCI patients. However, there is conflicting evidence regarding the effects of IH on memory function. The aim of this study was to assess episodic verbal and visual memory function with the Complutense verbal learning test (TAVEC) and the Rey-Osterrieth Complex Figure Test (ROCF), respectively, before and after a 4-week protocol of repetitive IH combined with body weight-supported treadmill training (BWSTT) in incomplete ASIA C and D SCI subjects. Subjects received either IH (cycling 9%/21% FiO2 every 1.5 min, 15 cycles per day) or continued normoxia (Nx, 21% FiO2) combined with 45 min of BWSTT for 5 consecutive days, followed by 3 times per week IH and BWSTT for 3 additional weeks. ROCF Z scores between IH plus BWSTT and Nx plus BWSTT were not significantly different (p = .43). Compared with baseline, IH and BWSTT group showed a significantly greater (p < .05) verbal memory performance for immediate, short-term, and long-term recall; however, it was not different from Nx plus BWSTT group in all verbal memory components (p > .05). Our results suggest that a 4-week protocol of moderate IH does not elicit visual or verbal memory impairment. Thus, repetitive IH may be a safe therapeutic approach to incomplete spinal cord injury patients, without deleterious cognitive effects.


[Effect of uremia and peritoneal dialysis on peritoneal mesothelial cells].
The aim of the study was to investigate the morphology of mesothelial cells of the peritoneum of patients with terminal renal failure (TRF), taken by the biopsy immediately before the onset of peritonal dialysis (PD), and to compare it with the findings in patients with PD. The samples were prepared in the way standard for light microscopy and transmission electron microscopy. In patients with TRF intracytoplasmatic inclusions could be observed, unusual protrusions of mesothelial apical surfaces, deformation of mesothelial cells and their detachment from the basal membrane, as well as the dilatated cisternae of granulated endoplasmatic reticulum with filamentous structures in some of them. In patients on PD cytoplasmic protrusions of different shapes and contents were observed at the surface of mesothelial cells, multiplication of basal membrane, occurrence of young forms of mesothelial cells as well as the detachment of those cells from the basal lamina.


Pregnant women with diabetes and their clinician's experience of participating in a pilot randomised controlled trial of corticosteroid administration in late pregnancy: A qualitative study.
Little research exists to support the administration of corticosteroids to pregnant women with diabetes. Pregnant women are often excluded from clinical trials due to concerns of harm to the foetus. This study aimed to understand the experiences of women and clinicians of participating in the Prevention of neonatal Respiratory distress with antenatal corticosteroids before Elective Caesarean section in women with Diabetes pilot randomised controlled trial to determine the acceptability of the study protocol. Women and clinicians participating in the pilot trial were invited to complete a telephone interview regarding their experiences of participating. Qualitative data were collected and subsequently analysed using thematic analysis. A total of 13 women and nine clinicians were recruited between June 2020 and May 2022 for a telephone interview. Participating in the study was deemed acceptable by women and clinicians. Women chose to participate in the study due to the perceived low risk of harm associated with the intervention and for altruistic reasons. The high level of clinical support and information provided for the duration of the pilot trial was valued by women and clinicians. All clinicians highlighted the importance of conducting the trial to inform evidence-based practice. Pregnant women are more likely to participate in clinical trials when perceived risks are low and they are well-informed during decision-making. Clinicians will support clinical trials when they perceive a benefit to practice and feel assured that women receive extensive monitoring and support. Incorporating these factors into study protocols is more likely to be successful in recruiting pregnant women and maintaining the engagement of clinical staff for the duration of clinical trials. Patients were invited to be participants in this study. A consumer has been included in the planning and oversite of the large multicentre trial.


Pulmonary involvement with Erdheim-Chester disease: radiographic and CT findings.
We determined the chest radiographic and CT findings of Erdheim-Chester disease with pulmonary involvement. We retrospectively reviewed the radiologic images of 15 patients with biopsy-proven Erdheim-Chester disease. Nine patients had chest radiographic images and CT scans that were available for review. Six men and three women were studied (age range, 25-70 years; mean age, 56 years). Two radiologists interpreted all images by consensus. Lung parenchyma was assessed for the type and distribution of disease. Bronchi, pleurae, hila, and mediastinal and extrathoracic structures were evaluated for abnormalities. Pathologic specimens from all patients were reviewed and correlated with radiologic findings. Eight of nine patients had thoracic images with abnormal findings. The most common radiographic pattern was reticular interstitial opacities with fissural and interlobular septal thickening. CT revealed regions of ground-glass attenuation and centrilobular nodular opacities. Typically, extrapulmonary findings included pleural effusions (6/8 patients), pericardial fluid or thickening (4/8), and extrathoracic infiltrative soft-tissue masses (4/8). The most common findings of Erdheim-Chester disease with pulmonary involvement include an interstitial process characterized by smooth interlobular septal thickening and centrilobular nodular opacities, fissural thickening, and pleural effusions. On CT, six of nine patients had pericardial fluid and thickening or extrathoracic soft-tissue masses. Such findings are characteristic of Erdheim-Chester disease with pulmonary involvement. Definitive diagnosis requires correlating skeletal findings and lung biopsy findings.


The Evolution of Nursing Research.
THE RESEARCH CULTURE in nursing has evolved in the last 150 years, beginning with Nightingale's work in the mid-1850s and culminating in the creation of the National Institute of Nursing Research (NINR) at the National Institues of Health (NIH). This article highlights nursing's efforts to facilitate the growth of the research culture by developing theory, establishing the importance of a research-based practice, advancing education, and providing avenues for dissemination of research. Similarities with the chiropractic profession are discussed, along with a commentary by Cheryl Hawk, D.C, Ph.D.


Analysis of a complex oligogenic disease.
Our analysis of GAW10 problem 2 data set consisted of linear regression analysis followed by linkage analysis. The linear regression analysis allowed some exploration of the relationships between the quantitative variables. Furthermore, it isolated some of the components of certain quantitative variables that were not due to a major locus and facilitated the linkage analysis that followed. For the linkage analysis, we used MAPMAKER/SIBS and the SIBPAL program from S.A.G.E. We found linkage of Q4 to chromosome 8. Analysis using the residuals of Q1 and Q3 showed linkage to chromosomes 5 and 4, respectively.


Molluscum contagiosum on tattoo.
Molluscum contagiosum is a disease caused by a poxvirus characterized by benign self-limited eruption of single or multiple cutaneous spherical and pearly papules. Transmission usually occurs by direct contact with infected hosts. It is reported the case of a 22-year-old Caucasian male who presented characteristic pearly and umbilicated papules strictly located on the region of a tattoo. Histopathologic exam confirmed the diagnosis of molluscum contagiosum. The authors describe an uncommomn manifestation of dissemination of this virus in tattoos and also present a literature review emphasizing the transmission pathways and treatment of Molluscum contagiosum.


An advantage for young sperm in the house cricket Acheta domesticus.
We show that males of the house cricket Acheta domesticus regularly expel sperm packages (spermatophores) independently of copulation and at a rate that is not affected by the presence of females. We then show for the first time that the age of sperm affects their likelihood of being stored by females after copulation; younger sperm were overrepresented in the female sperm storage organ and therefore in the sperm population used for fertilization. Our results suggest that the reproductive success of males may increase if they deliver ejaculates with young sperm, and the results may explain why the males of several species are regularly observed to discard ejaculates. Our results also suggest that phenomena such as female multiple mating, paternity bias, and/or exaggerated ejaculate sizes may be related to the advantage both genders gain by using young sperm.


Short-course high-dose dexamethasone therapy for chronic idiopathic thrombocytopenic purpura in children.
First-line therapies of acute and chronic idiopathic thrombocytopenic purpura (ITP) include intravenous immunoglobulin, IV anti-D and corticosteroids. A short-course high-dose dexamethasone (HDD-SC) therapy has recently been reported to be efficacious in acute ITP. The present study was conducted to assess the efficacy of HDD-SC in children with chronic ITP. Over a period of 10 months, 13 patients with chronic ITP were given HDD-SC (20 mg m(-2) IV daily for 4 days, four cycles repeated every 15 days). Of the 12 patients who could be evaluated, complete response was observed in 8 (66.6%) and moderate response in 2 (17%) patients, whereas 2 (17%) patients had no response. HDD-SC appears to be a safe and effective therapy in childhood ITP.


Consensus agreement to rename burning mouth syndrome and improve International Classification of Diseases-11 disease criteria: an international Delphi study.
The International Classification of Diseases (ICD-11) proposes revisions in the nomenclature, disease definition, and diagnostic criteria for "burning mouth syndrome" (BMS). This process could benefit from additional systematically collected expert input. Thus, the purpose of this study was to use the Delphi method to (1) determine whether revision in nomenclature and alternative names for "BMS" are warranted and (2) identify areas of consensus among experts for changes to the disease description and proposed diagnostic criteria of "BMS," as described in the ICD-11 (World Health Organization). From 31 international invited experts, 23 who expressed interest were sent the survey. The study used 4 iterative surveys, each with a response rate of ≥82%. Consensus was predefined as 70% of participants in agreement. Data were summarized using both descriptive statistics and qualitative thematic analysis. Consensus indicated that BMS should not be classified as a syndrome and recommended instead renaming to "burning mouth disorder." Consensus included deletion of 2 diagnostic criteria: (1) emotional distress or functional disability and (2) the number of hours symptoms occur per day. Additional items that reached consensus clarified the disease definition and proposed more separate diagnostic criteria, including a list of local and systemic factors to evaluate as potential secondary causes of oral burning. Experts in this study recommended and came to consensus on select revisions to the proposed ICD-11 BMS nomenclature, diagnostic criteria, and disease definition. The revisions recommended have the potential to improve clarity, consistency, and accuracy of diagnosis for this disorder.


Prevalence of rat virus infection in progeny of acutely or persistently infected pregnant rats.
Infant rats are susceptible to persistent rat virus (RV) infection, but risk of persistent infection after prenatal exposure to virus is unclear. We examined this aspect of RV infection in the progeny of dams inoculated with virus during or prior to pregnancy. Sprague-Dawley (SD) dams were infected during pregnancy (gestation day 9) by oronasal inoculation with 10(5) TCID50 of the UMass strain of RV. SD rats were infected prior to pregnancy by oronasal inoculation of two-day-old females with 10(2) TCID50 of RV-UMass, which induced persistent infection. They were mated to non-immune males after reaching sexual maturity. Rats were assessed for RV infection by virus isolation, in situ hybridization, contact transmission, or serologic testing. The progeny of dams inoculated with virus during gestation had high prevalence of infection through postpartum week 9 (9 of 12 rats were virus positive at week 3, and 7 of 10 were virus positive at week 9). Additionally, 2 of 10 rats were virus positive at least through postpartum week 15. The progeny from persistently infected, seropositive dams had no evidence of infection and did not transmit infection to contact sentinels. However, 12 dams were virus positive at necropsy and 9 had transmitted infection to their breeding partners. These results indicate that prenatal infection in non-immune dams can lead to RV persistence in their progeny. By contrast, the progeny of persistently infected dams are protected from infection, presumably by maternal antibody, although their dams can transmit infection to their breeding partners.


Association of Torquetenovirus Viremia with Physical Frailty and Cognitive Impairment in Three Independent European Cohorts.
Immunosenescence and inflammaging have been implicated in the pathophysiology of frailty. Torquetenovirus (TTV), a single-stranded DNA anellovirus, the major component of the human blood virome, shows an increased replication rate with advancing age. An elevated TTV viremia has been associated with an impaired immune function and an increased risk of mortality in the older population. The objective of this study was to analyze the relation between TTV viremia, physical frailty, and cognitive impairment. TTV viremia was measured in 1,131 nonfrail, 45 physically frail, and 113 cognitively impaired older adults recruited in the MARK-AGE study (overall mean age 64.7 ± 5.9 years), and then the results were checked in two other independent cohorts from Spain and Portugal, including 126 frail, 252 prefrail, and 141 nonfrail individuals (overall mean age: 77.5 ± 8.3 years). TTV viremia ≥4log was associated with physical frailty (OR: 4.69; 95% CI: 2.06-10.67, p < 0.0001) and cognitive impairment (OR: 3.49, 95% CI: 2.14-5.69, p < 0.0001) in the MARK-AGE population. The association between TTV DNA load and frailty status was confirmed in the Spanish cohort, while a slight association with cognitive impairment was observed (OR: 1.33; 95% CI: 1.000-1.773), only in the unadjusted model. No association between TTV load and frailty or cognitive impairment was found in the Portuguese sample, although a negative association between TTV viremia and MMSE score was observed in Spanish and Portuguese females. These findings demonstrate an association between TTV viremia and physical frailty, while the association with cognitive impairment was observed only in the younger population from the MARK-AGE study. Further research is necessary to clarify TTV's clinical relevance in the onset and progression of frailty and cognitive decline in older individuals.


Knowledge and perceptions of quality of obstetric and newborn care of local health providers: a cross-sectional study in three districts in Malawi.
Quality of service delivery for maternal and newborn health in Malawi is influenced by human resource shortages and knowledge and care practices of the existing service providers. We assessed Malawian healthcare providers' knowledge of management of routine labour, emergency obstetric care and emergency newborn care; correlated knowledge with reported confidence and previous study or training; and measured perception of the care they provided. This study formed part of a large-scale quality of care assessment in three districts (Kasungu, Lilongwe and Salima) of Malawi. Subjects were selected purposively by their role as providers of obstetric and newborn care during routine visits to health facilities by a research assistant. Research assistants introduced and supervised the self-completed questionnaire by the service providers. Respondents included 42 nurse midwives, 1 clinical officer, 4 medical assistants and 5 other staff. Of these, 37 were staff working in facilities providing Basic Emergency Obstetric Care (BEMoC) and 15 were from staff working in facilities providing Comprehensive Emergency Obstetric Care (CEMoC). Knowledge regarding management of routine labour was good (80% correct responses), but knowledge of correct monitoring during routine labour (35% correct) was not in keeping with internationally recognized good practice. Questions regarding emergency obstetric care were answered correctly by 70% of respondents with significant variation depending on clinicians' place of work. Knowledge of emergency newborn care was poor across all groups surveyed with 58% correct responses and high rates of potentially life-threatening responses from BEmOC facilities. Reported confidence and training had little impact on levels of knowledge. Staff in general reported perception of poor quality of care. Serious deficiencies in providers' knowledge regarding monitoring during routine labour and management of emergency newborn care were documented. These may contribute to maternal and neonatal deaths in Malawi. The knowledge gap cannot be overcome by simply providing more training.


The way forward in reproductive/developmental toxicity testing.
The use of experimental animals in reproductive toxicity testing is critically reviewed on the occasion of the 50th anniversary of the publication of the Three Rs concept by Russell and Burch, since there is major concern that reproductive toxicity testing will significantly increase due to the requirements of the EU Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH) system. A comparison of the test guidelines for drugs, agrochemicals and industrial chemicals shows that, for historical reasons, significantly different testing strategies are applied. The current status of development and validation of in vitro tests in reproductive toxicology is also critically evaluated. The mouse embryonic stem cell test (mEST) is the most advanced and promising of the in vitro tests. Although it has not yet been accepted for regulatory purposes, its use in preclinical drug development is well established. Moreover, promising molecular endpoints have been established in the mEST, including proteomic and toxicogenomic endpoints. Preliminary results have been obtained with a human EST (hEST). In addition, an overview is given on new in vitro reproductive toxicity tests that are currently being developed in the EU FP6 project, ReProTect, since the ReProTect test battery, which covers the essential steps of female and male fertility, implantation and embryotoxicity, holds promise for use as a screening assay for reproductive toxicity testing according to the EU REACH legislation. However, since validated in vitro methods will not be available in the short term, opportunities for the refinement of the standard in vivo tests are discussed, in order to reduce the numbers of animal used in reproductive toxicity testing. Finally, recommendations for toxicity testing in the 21st century call for the harmonisation of test methods across all areas of regulatory testing as a first step. Since the REACH system testing framework for industrial chemicals is driven by the reproductive safety testing requirements of agrochemicals, a shift is proposed to exposure-driven testing of industrial chemicals. In particular, the implementation of a new 'extended one-generation reproductive toxicity study' (EOGRTS), which includes triggers for additional testing for fertility, developmental neurotoxicity and immunotoxicity, would significantly reduce test animal numbers. It is concluded that in vitro methods hold great promise for reproductive toxicity testing in the 21st century, e.g. the ReProTect in vitro battery and the embryonic stem cell (ESC) technology focusing on molecular endpoints in both the mEST and the hEST.


Comparative performance of the Roche COBAS Amplicor assay and an in-house real-time PCR assay for diagnosis of Chlamydia trachomatis infection.
This study investigated the comparative performance of the Amplicor assay and an in-house semi-automated, multiplex real-time PCR for the diagnosis of genital chlamydial infection. Four different assays, the COBAS Amplicor CT test (Amplicor PCR), in-house real-time PCR (IHRT-PCR), in-house nested cryptic plasmid PCR and in-house nested major outer membrane protein PCR, were performed on genital swabs from 1000 consecutive patients attending a genitourinary medicine clinic. The samples were designated true positive if Chlamydia trachomatis DNA was detected by at least two of the four above-mentioned assays while a sample was defined as true negative if C. trachomatis DNA was detected in only one or none of the assays. By this criterion, there were 129 true positive and 871 true negative samples for C. trachomatis DNA in this cohort. Amplicor PCR designated 144 samples positive: 128 (89%) of 144 samples were true positive and 16 (11%) were false positive. IHRT-PCR detected 126 of 129 true positive samples and did not generate any false positive results. The sensitivity of IHRT-PCR was comparable with, and specificity was higher than, Amplicor PCR for the diagnosis of genital chlamydial infection.


Numerical simulation of transient dynamic behavior of healthy and hardened red blood cells in microcapillary flow.
In a number of human diseases such as diabetes mellitus and sickle cell anemia, variations in mechanical properties of red blood cells (RBCs) occur and cause reduced deformability. Investigating the behavior of such abnormal, hardened RBCs in microcapillary flow is of prime importance because of their effects on oxygen transport process. In the present paper, dynamic response of a RBC to a microcapillary flow is numerically studied at steady and transient conditions, considering the effect of essential parameters including RBC deformability, its initial orientation, velocity, and flow pressure gradient. Simulations are performed using a three-dimensional hybrid method, combining lattice Boltzmann method for plasma flow, finite element method for RBC membrane analysis, and immersed boundary method for their interaction. Quantitative and qualitative validations with the experimental data for different RBC velocities verify the accuracy of applied numerical method. Apart from the initial orientation, RBC experiences a complex shape deformation in which the biconcave discoid shape changes to a parachute-like shape. While deformation index of RBC does not change considerably with RBC deformability at steady state condition, it plays an important role in its shape evolution under transient condition. Copyright © 2016 John Wiley & Sons, Ltd.


Momentum injection in tokamak plasmas and transitions to reduced transport.
The effect of momentum injection on the temperature gradient in tokamak plasmas is studied. A plausible scenario for transitions to reduced transport regimes is proposed. The transition happens when there is sufficient momentum input so that the velocity shear can suppress or reduce the turbulence. However, it is possible to drive too much velocity shear and rekindle the turbulent transport. The optimal level of momentum injection is determined. The reduction in transport is maximized in the regions of low or zero magnetic shear.


Allograft replacement of the aortic valve for active endocarditis.
In the 50 months preceding March 1995, 35 adult patients underwent aortic valve replacement with a cryopreserved human aortic valve allograft. Nine of these patients had active endocarditis. The remaining 26 patients had aortic valve pathology without active infection. This study consists of a comparison of these two groups. One of the patients with active infection underwent inclusion technique replacement of the infected valve. The other eight valves were replaced with root-replacement technique. Of the 26 non-infected patients, 4 valves were replaced with the inclusion technique, and the remaining 22 were replaced with the root-replacement technique. None of the patients are maintained on long-term anti-coagulation. We have followed the infected group an average of 32 months and the group without active infection has been followed an average of 27 months. There were two perioperative deaths in the nine patients with infected valves and no deaths in the group without active infection (Pearson's chi2 p<0.05). There have been no thromboembolic complications and no recurrent infections.


In vivo antioxidant activity of Okara Koji, a fermented okara, by Aspergillus oryzae.
The antioxidants in Okara Koji (OK), an okara (OC) fermented by Aspergillus oryzae, gamma-tocopherol, delta-tocopherol, genistin, daizein, genistein, and 3-hydroxyanthranilic acid were identified by HPLC. OK's extract with 80% methanol strongly inhibited linoleate peroxidation, much more than other OK's extracts with hexane or hot water. The methanol extract accelerated 12-hydroxyeicosatetraenoic acid formation in membrane lipids at 10(-3) concentration, but inhibited the formation at higher concentrations than 10(-3) ex vivo. To confirm the total effect of all components of OK on lipid peroxidation in vivo, rats fed food deficient in vitamin E were put on diets containing OK or OC with oxidized oil. In rats fed the OK diet, no effect of oxidized oil feeding on the body weight gain, of the TBA value in plasma, or of glutathione peroxidase activities of plasma and liver was observed. But in rats fed the OC diet, the effect of oxidized oil feeding was apparently observed on all of those values. These results suggested that OK would scavenge lipid peroxides in vivo.


Neural Network Differential Equations For Ion Channel Modelling.
Mathematical models of cardiac ion channels have been widely used to study and predict the behaviour of ion currents. Typically models are built using biophysically-based mechanistic principles such as Hodgkin-Huxley or Markov state transitions. These models provide an abstract description of the underlying conformational changes of the ion channels. However, due to the abstracted conformation states and assumptions for the rates of transition between them, there are differences between the models and reality-termed model discrepancy or misspecification. In this paper, we demonstrate the feasibility of using a mechanistically-inspired neural network differential equation model, a hybrid non-parametric model, to model ion channel kinetics. We apply it to the hERG potassium ion channel as an example, with the aim of providing an alternative modelling approach that could alleviate certain limitations of the traditional approach. We compare and discuss multiple ways of using a neural network to approximate extra hidden states or alternative transition rates. In particular we assess their ability to learn the missing dynamics, and ask whether we can use these models to handle model discrepancy. Finally, we discuss the practicality and limitations of using neural networks and their potential applications.


Pulsed Nd:YAG laser: effects on pain, postural stability, and weight-bearing pattern in children with hemophilic ankle arthropathy.
Hemophilic ankle arthropathy features joint pain, impaired postural control, and abnormal weight-bearing pattern. This study was designed to investigate the effects of pulsed Nd:YAG laser on pain, postural stability, and weight-bearing pattern in children with hemophilic ankle arthropathy. Forty children diagnosed with hemophilia type A (age, 8-16 years) were randomly allocated to either the treatment group (n = 20) who received a three-phase active therapy with pulsed Nd:YAG laser thrice/week (total energy was 1500 J) plus a physical exercise program for four consecutive weeks, or the placebo group (n = 20) who received placebo laser plus the physical exercise program. Pain, postural control (i.e., directional control [DC], endpoint excursion [EE], center-of-gravity movement velocity [CoG-MV], and maximum excursion [MXE]), and weight-bearing pattern (i.e., pressure peaks of the hindfoot [PP-HF] and forefoot [PP-FF], and the foot contact area [FCA]) were assessed pre- and post-treatment. The treatment group showed greater improvement in pain (P = .004), DC (P = .02), EE (P = .003), and CoG-MV (P = .003) compared to the placebo group. However, there was no statistically significant difference between both groups regarding the MXE (P = .15). In addition, the treatment group achieved favorable improvements in PP-HF (P = .003), PP-FF (P = .027), and FCA (P = .002) relative to the placebo group. Pulsed Nd:YAG laser is a potentially effective therapy for pain relief, postural control enhancement, and weight-bearing pattern adjustment in children with hemophilic ankle arthropathy.


How ecological research on human-dominated ecosystems incorporates agricultural and forestry practices: A literature analysis.
Understanding the nexus between practices and ecosystems is a critical challenge for sustainability, but it is unclear how ecological sciences have explored the question. To bring clarification, we conducted an analysis of ecology literature dealing with agricultural and forestry practices (AF practices), scanning a total of 27 556 references. Scientometric analyses showed that AF practices were addressed by 5.5% of ecology literature, and that this proportion increased from 2.5 to 8.1% between 1956 and 2017. Content analyses showed that research has mainly focused on monospecific systems in the Global North, predominantly using plot-level experimental approaches. Temporal monitoring, real-world practices and their social context were poorly investigated. This analysis stresses the need to reinforce research in complex agroecosystems, in particular in non-Western countries. Multilevel and spatio-temporal approaches, as well as participatory research, should also be encouraged to build a social-ecological understanding and formulate more grounded, relevant policy recommendations for sustainability.


Mendelian randomization reveals association between retinal thickness and non-motor symptoms of Parkinson's disease.
Retinal thickness is related to Parkinson's disease (PD), but its association with the severity of PD is still unclear. We conducted a Mendelian randomized (MR) study to explore the association between retinal thickness and PD. For the two-sample MR analysis, the summary statistics obtained from genome-wide association studies on the thickness of Retinal nerve fiber layer (RNFL) and ganglion cell inner plexiform layer (GCIPL) were employed as exposure, while the summary statistics associated with PD were used as the outcome. The primary approach utilized was inverse variance weighted. To correct for multiple testing, the false discovery rate (FDR) was employed. For sensitivity analysis, an array of robust MR methods was utilized. We found genetically predicted significant association between reduced RNFL thickness and a reduced risk of constipation in PD (odds ratio [OR] = 0.854, 95% confidence interval [CI] (0.782, 0.933), P < 0.001, FDR-corrected P = 0.018). Genetically predicted reduced RNFL thickness was associated with a reduced Unified Parkinson's Disease Rating Scale total score (β = -0.042, 95% CI (-0.079, 0.005), P = 0.025), and reduced GCIPL thickness was associated with a lower risk of constipation (OR = 0.901, 95% CI (0.821, 0.988), P = 0.027) but a higher risk of depression (OR = 1.103, 95% CI (1.016, 1.198), P = 0.020), insomnia (OR = 1.090, 95% CI (1.013, 1.172), P = 0.021), and rapid eye movement sleep behaviour disorder (RBD) (OR = 1.198, 95% CI (1.061, 1.352), P = 0.003). In conclusion, we identify an association between retinal thickness and non-motor symptoms (constipation, depression, insomnia and RBD) in PD, highlighting the potential of retinal thickness as a biomarker for PD nonmotor symptoms.


Multi-agent cooperative target search.
This paper addresses a vision-based cooperative search for multiple mobile ground targets by a group of unmanned aerial vehicles (UAVs) with limited sensing and communication capabilities. The airborne camera on each UAV has a limited field of view and its target discriminability varies as a function of altitude. First, by dividing the whole surveillance region into cells, a probability map can be formed for each UAV indicating the probability of target existence within each cell. Then, we propose a distributed probability map updating model which includes the fusion of measurement information, information sharing among neighboring agents, information decay and transmission due to environmental changes such as the target movement. Furthermore, we formulate the target search problem as a multi-agent cooperative coverage control problem by optimizing the collective coverage area and the detection performance. The proposed map updating model and the cooperative control scheme are distributed, i.e., assuming that each agent only communicates with its neighbors within its communication range. Finally, the effectiveness of the proposed algorithms is illustrated by simulation.


[Prevention and treatment of early complications in patients after reconstructive surgery of the esophagus].
The work deals with clinical appraisal of the course of the early postoperative period and the prevention and treatment of complications after reconstructive operations on the esophagus. The postoperative period was studied in 99 patients. Complications occurred in the early postoperative period in 48.5% of patients, obstructive processes in the lungs were of principal importance among them. The authors conclude that particular significance in the prevention of acute respiratory failure in patients after one-stage plasty of the esophagus should be attached to the early diagnosis of pulmonary complications, prolonged artificial lung respiration, and measures for maintaining normal passage of the respiratory tract (cleansing bronchoscopy, early activation of patients, adequate anesthesia). The main importance in reducing mortality after one-stage esophageal plasty belongs to the measures of prevention and treatment of incompetence of the sutures of the esophageal anastomosis and mediastinitis.


Higher farnesyl diphosphate synthase activity in human colorectal cancer inhibition of cellular apoptosis.
Farnesyl diphosphate synthase (FPPs) produces FPP which is considered a branch-point intermediate in the synthesis of sterols and isoprenylated cellular metabolites. In this study we investigated whether detectable FPPs activity was present in human colorectal cancer (CRC), also evaluating in vitro the role of this enzyme in the growth and apoptosis of CRC cells by using Pamidronate (PAM), a FPPs activity inhibitor. The activity level of FPPs was determined in CRC and the normal surrounding mucosa of 50 patients by radiochemical assay. The FPPs mRNA expression was investigated in 15 of 50 patients by quantitative reverse transcriptase polymerase chain reaction (RT-PCR). K-ras mutation was evaluated using PCR and restriction enzyme analysis. Cell growth and apoptosis, after PAM treatment, in human CRC cell line DLD-1 were measured by MTT test and DNA fragmentation, respectively. FPPs activity was detectable in human CRC. FPPs activity and its mRNA were significantly more abundant in cancer samples than in normal mucosa. In vitro PAM resulted in a significant reduction of cell growth and also gave rise to a marked proapoptotic effect. This study provides the first evidence of the presence of FPPs activity in human CRC. Moreover, FPPs enzyme was found to play a significant role in colon cancer proliferation.


[Acupuncture analgesia in China (author's transl)].
As part of an agreement between the Governments of the People's Republic of China and Canada, 10 Canadian Anaesthetists visited China for six weeks during April and May 1974. The delegation observed 87 surgical operations and 19 dental procedures conducted under acupuncture analgesia in 17 large hospital in Peking, Schichiachuang, Nanking, Shanghai and Canton. In this report, observations on acupuncture analgesia made during this visit are presented as well as information obtained in group discussions with Chinese physicians. In the author's opinion, the technique was successful in about 80% of the cases seen; these patients were awake during surgery without visible distress and exhibited a remarkable degree of analgesia immediately after surgery. However, in China, acupuncture analgesia is still considered in the experimental stage and only used in a limited number of standardized, elective surgical procedures, where confidence and the success of this technique have been established. For the successful use of this technique, patient and surgeon must co-operate fully. Acupuncture analgesia poses considerable limitations on the surgeon and requires careful surgical technique; the patient's co-operation must be assured by a suitable preparation. It appears, that acupuncture analgesia should be seen as an alternative to local anaesthesia rather than general anaesthesia; the latter is only used to a limited extent in China. In our present anaesthetic practice, acupuncture would appear to be of only very limited use, however, a further study of this phenomenon should contribute to our knowledge of the function of the nervous system.


Synthesis and Photophysical Properties of 3-Substituted-1H-Indazoles: A Pd-Catalyzed Double C-N Bond Formation Strategy via 1,6-Conjugate Addition.
A Pd-catalyzed cascade process for the direct synthesis of 3-substituted-1H-indazole employing p-quinone methide (p-QM) and arylhydrazine through Pd-catalyzed double C-N bond formation via 1,6-conjugate addition is reported. This reaction strategy affords efficient and practical access to synthetically important diverse 3-substituted-1H-indazoles in good yields. The photophysical properties of the synthesized 3-substituted-1H-indazoles are investigated, and some of them showed very good fluorescence properties with quantum yields up to 85%. Also, the synthesized 3-substituted-1H-indazole exhibits an acid-sensitive fluorescence turn-off activity.


Inter-rater reliability of two paediatric early warning score tools.
Paediatric early warning score (PEWS) assessment tools can assist healthcare providers in the timely detection and recognition of subtle patient condition changes signalling clinical deterioration. However, PEWS tools instrument data are only as reliable and accurate as the caregivers who obtain and document the parameters. The aim of this study is to evaluate inter-rater reliability among nurses using PEWS systems. The study was carried out in five paediatrics departments in the Central Denmark Region. Inter-rater reliability was investigated through parallel observations. A total of 108 children and 69 nurses participated. Two nurses simultaneously performed a PEWS assessment on the same patient. Before the assessment, the two participating nurses drew lots to decide who would be the active observer. Intraclass correlation coefficient, Fleiss' κ and Bland-Altman limits of agreement were used to determine inter-rater reliability. The intraclass correlation coefficients for the aggregated PEWS score of the two PEWS models were 0.98 and 0.95, respectively. The κ value on the individual PEWS measurements ranged from 0.70 to 1.0, indicating good to very good agreement. The nurses assigned the exact same aggregated score for both PEWS models in 76% of the cases. In 98% of the PEWS assessments, the aggregated PEWS scores assigned by the nurses were equal to or below 1 point in both models. The study showed good to very good inter-rater reliability in the two PEWS models used in the Central Denmark Region.


A new family of 10 murine ovalbumin serpins includes two homologs of proteinase inhibitor 8 and two homologs of the granzyme B inhibitor (proteinase inhibitor 9).
Serine proteinase inhibitors (serpins) are classically regulators of extracellular proteolysis, however, recent evidence suggests that some function intracellularly. Such "ovalbumin" serpins include the human proteinase inhibitors 6 (PI-6), 8 (PI-8), and 9 (PI-9), plasminogen activator inhibitor 2, and the monocyte/neutrophil elastase inhibitor. PI-9 is a potent granzyme B (graB) inhibitor that has an unusual P1 Glu and is present primarily in lymphocytes. In a search for the murine equivalent of PI-9 we screened cDNA libraries, and performed reverse transcriptase-polymerase chain reaction on RNA isolated from leukocyte cell lines and from lymph nodes and spleens of allo-immunized mice. We identified 10 new ovalbumin serpin sequences: two resemble PI-8, two resemble PI-9, and the remaining six have no obvious human counterparts. By RNA analysis only one of the two sequences resembling PI-9 (designated SPI6) is present in mouse lymphocytes while the other (a partial clone designated mBM2A) is predominantly in testis. SPI6 comprises a 1.8-kilobase cDNA encoding a 374-amino acid polypeptide that is 68% identical to PI-9. mBM2A is 65% identical to PI-9 and over 80% identical to SPI6. Although the reactive loops of SPI6 and mBM2A differ from PI-9, both contain a Glu in a region likely to contain the P1-P1' bond. SPI6 produced in vitro using a coupled transcription/translation system formed an SDS-stable complex with human graB and did not interact with trypsin, chymotrypsin, leukocyte elastase, pancreatic elastase, thrombin, or cathepsin G. Recombinant SPI6 produced in a yeast expression system was used to examine the interaction with human graB in more detail. The second-order rate constant for the interaction was estimated as 8 x 10(4) M-1 s-1, and inhibition depended on the Glu in the SPI6 reactive center. The SPI6 gene was mapped to the same region on mouse chromosome 13 as Spi3, which encodes the murine homolog of PI-6. We conclude that even though their reactive centers are not highly conserved, SPI6 is a functional homolog of PI-9, and that the regulation of graB in the mouse may involve a second serpin encoded by mBM2A. Our identification of multiple sequence homologs of PI-8 and PI-9, and six new ovalbumin serpins, is consonant with the idea that the larger set of granule and other proteinases known to exist in the mouse (compared with human) is balanced by a larger array of serpins.


Combining resonance energy transfer methods reveals a complex between the alpha2A-adrenergic receptor, Galphai1beta1gamma2, and GRK2.
Traditionally, G-protein-coupled receptor (GPCR) interactions with their G proteins and regulatory proteins, GPCR kinases (GRKs) and arrestins, are described as sequential events involving rapid assemblies/disassemblies. To directly monitor the dynamics of these interactions in living cells, we combined two spectrally resolved bioluminescence and one fluorescence resonance energy transfer (RET) methods. The RET combination analysis revealed that stimulation of the α(2A)-adrenergic receptor (α(2A)AR) leads to the recruitment of GRK2 at a receptor still associated with the Gα(i1)β(1)γ(2) complex. The interaction kinetics of GRKs with Gγ(2) (2.8 ± 0.4 s) and α(2A)AR (5.2 ± 0.5 s) were similar to that of the receptor-promoted change in RET between Gα(i1) and Gγ(2) (5.2 ± 1.2 s), and persisted until the translocation of βarrestin2 to the receptor, indicating that GRK2 remains associated to the receptor/G-protein complex for longer periods than anticipated. Moreover, GRK2 or a kinase-deficient GRK2 mutant, but not GRK5, potentiated the receptor-promoted changes in RET between Gα(i1) and Gγ(2) and abrogated the α(2A)AR-stimulated calcium response, suggesting that the recruitment of GRK2 to the complex contributes to the structural rearrangement and functional regulation of the signaling unit, independently of the kinase activity. RET combination analysis revealed unanticipated dynamics in GPCR signaling and will be applicable to many biological systems.


Electrical Stimulation of Artificial Heart Muscle: A Look Into the Electrophysiologic and Genetic Implications.
Development of tissue-engineered hearts for treatment of myocardial infarction or biologic pacemakers has been hindered by the production of mostly arrhythmic or in-synergistic constructs. Electrical stimulation (ES) of these constructs has been shown to produce tissues with greater twitch force and better adrenergic response. To further our understanding of the mechanisms underlying the effect of ES, we fabricated a bioreactor capable of delivering continuous or intermittent waveforms of various types to multiple constructs simultaneously. In this study, we examined the effect of an intermittent biphasic square wave on our artificial heart muscle (AHM) composed of neonatal rat cardiac cells and fibrin gel. Twitch forces, spontaneous contraction rates, biopotentials, gene expression profiles, and histologic observations were examined for the ES protocol over a 12 day culture period. We demonstrate improved consistency between samples for twitch force and contraction rate, and higher normalized twitch force amplitudes for electrically stimulated AHMs. Improvements in electrophysiology within the AHM were noted by higher conduction velocities and lower latency in electrical response for electrically stimulated AHMs. Genes expressing key electrophysiologic and structural markers peaked at days 6 and 8 of culture, only a few days after the initiation of ES. These results may be used for optimization strategies to establish protocols for producing AHMs capable of replacing damaged heart tissue in either a contractile or electrophysiologic capacity. Optimized AHMs can lead to alternative treatments to heart failure and alleviate the limited donor supply crisis.