Gastrostomy-to-gastrojejunostomy tube conversion: impact of the method of original gastrostomy tube placement.
To determine the outcome of gastrostomy tube-to-gastrojejunostomy tube conversion on the basis of the method of original gastrostomy tube placement. One hundred twenty-four patients (age range, 13-87 years; 72 male and 52 female patients) underwent conversion of a primarily placed gastrostomy tube to a gastrojejunostomy tube at the authors' institution between January 2000 and December 2008. The method of original gastrostomy tube placement was radiologic (n = 27), endoscopic (n = 75), laparoscopic (n = 2), or open surgery (n = 20). The method of placement was correlated with the success rates of gastrostomy-to-gastrojejunostomy tube conversion. Medical records and radiologic images were reviewed to determine the frequency of proximal migration of the jejunostomy tube into the stomach. Follow-up data were available for an average of 136 days after gastrostomy-to-gastrojejunostomy tube conversion (median, 63 days; range, 1-1,300 days). Of 124 gastrostomy tube-to-gastrojejunostomy tube conversions, 109 (87.9%) were successfully performed. Procedural conversion failure occurred in one of the 27 radiologically inserted gastrostomy tubes (3.7%) compared to 14 of the 97 (14%) nonradiologically inserted gastrostomy tubes (P = .19), of which 12 were inserted endoscopically and two were inserted surgically. Of the 109 patients with successful tube conversion, jejunal tip malposition occurred at follow-up in 18 (16.5%). Of these, four patients developed aspiration pneumonia (22%), which contributed to patient death in two. The frequency of jejunal tip malposition was 3.8% (one of 26 patients) for radiologically placed gastrostomy tubes and 20% (17 of 83 patients) for nonradiologically placed gastrostomy tubes (P = .07). Combined, 32% of gastrostomy tubes placed nonradiologically resulted in either procedural failure or eventual jejunal tip malposition, compared to 7.4% of radiologically placed gastrostomy tubes (P = .01). The frequency of procedural failure or eventual jejunal tip malposition with conversion of radiologically placed gastrostomy tubes to gastrojejunostomy tubes is significantly lower with radiologically placed gastrostomy tubes than with nonradiologically inserted gastrostomy tubes.


Modulatory effect of fluoride and irradiation on rat molar rate of wear.
The hypothesis was tested that fluoride (F-) modulates molar wear rate in the irradiated rat and that enamel solubility and dentin hardness are involved in this process. Seventy five 21 day-old rats were divided into 5 groups. Groups received either F-(25 ppm) in the drinking water or irradiation to the head (15 Gy in a single dose), or a combination of the two. The rate of occlusal wear was assessed by computerized planimetry. The amount of wear was significantly higher in the F- and irradiation monotreated rats, while under combined treatment it did nor differ significantly from the control values. Fluoridation or irradiation suppressed enamel solubility, as measured by calcium release in the etchant. Dentin microhardness, expressed in Vickers hardness number, was enhanced after either treatment, but remained unaffected when F- administration preceded irradiation. Enamel solubility and dentin microhardness did not correlate significantly with the rate of occlusal wear.


Prolonged pulseless perfusion in unanesthetized calves.
Nonpulsatile left ventricular (LV) bypass was achieved in nine alert, active calves for periods of up to two weeks using a centrifugal blood pump. Data were compared to prebypass control values and to results obtained from seven calves that underwent nonthoracotomy, retrograde LV cannulation, and pulsatile bypass for up to 35 days. Nonpulsatile flow produced an increase in heart rate of 20% (P less than .05) and an increase in aortic pressure of 10% (P less than .05). Cardiac output and systemic vascular resistance were similar in both groups for three days; thereafter, vascular resistance decreased as cardiac output progressively increased. Serum renin, lactate, and pyruvate values remained normal; creatinine clearances were not significantly different than control values, and there were no significant changes in body weight. These data indicate no untoward effects as a result of prolonged pulseless perfusion.


Controlled overexpression of Pax6 in vivo negatively autoregulates the Pax6 locus, causing cell-autonomous defects of late cortical progenitor proliferation with little effect on cortical arealization.
Levels of expression of the transcription factor Pax6 vary throughout corticogenesis in a rostro-lateral(high) to caudo-medial(low) gradient across the cortical proliferative zone. Previous loss-of-function studies have indicated that Pax6 is required for normal cortical progenitor proliferation, neuronal differentiation, cortical lamination and cortical arealization, but whether and how its level of expression affects its function is unclear. We studied the developing cortex of PAX77 YAC transgenic mice carrying several copies of the human PAX6 locus with its full complement of regulatory regions. We found that PAX77 embryos express Pax6 in a normal spatial pattern, with levels up to three times higher than wild type. By crossing PAX77 mice with a new YAC transgenic line that reports Pax6 expression (DTy54), we showed that increased expression is limited by negative autoregulation. Increased expression reduces proliferation of late cortical progenitors specifically, and analysis of PAX77<---->wild-type chimeras indicates that the defect is cell autonomous. We analyzed cortical arealization in PAX77 mice and found that, whereas the loss of Pax6 shifts caudal cortical areas rostrally, Pax6 overexpression at levels predicted to shift rostral areas caudally has very little effect. These findings indicate that Pax6 levels are stabilized by autoregulation, that the proliferation of cortical progenitors is sensitive to altered Pax6 levels and that cortical arealization is not.


Bridging the Gap: A Priorities Assessment Tool to Support Shared Decision Making, Maximize Appointment Time, and Increase Patient Satisfaction in Women With Ovarian Cancer.
Women with ovarian cancer identify patient-physician communication as an essential element in determining treatment course and believe a discussion about goals and values should precede treatment decisions. We sought to develop a patient-centered priorities assessment tool for women with ovarian cancer that could streamline communication, enhance treatment discussions, and increase patient satisfaction. We designed a priorities assessment tool using a validated ovarian cancer symptom index (National Comprehensive Cancer Center-Functional Assessment of Cancer Therapy Ovarian Symptom Index-18) combined with an index to assess daily quality-of-life priorities. The tool was distributed to women with ovarian cancer in small focus group settings and online, followed by a postactivity feedback form. In this pilot study, 36 women completed the priorities assessment tool and 35 completed the postactivity feedback form between September 2015 and May 2016. All participants reported that the tool was easy to understand and comprehensive in scope. Twenty-nine participants (82.9%) completed the tool in 10 minutes or less. Most participants (n = 31, 86.1%) were able to stratify their priorities and identify 5 top treatment-related priorities. Participants who indicated that their goals and priorities had changed since diagnosis (n = 25, 69.4%) reported that the tool helped to identify current goals and priorities (22 [88%] of 25 participants) and would help them feel more comfortable participating in shared decision making with their medical team (21 [84%] of 25 participants). A patient-centered priorities assessment tool was easy to complete and viewed as comprehensive and useful in a pilot cohort of women with ovarian cancer. Use of a priorities assessment tool has the potential to enhance communication, promote shared decision making, and improve patient satisfaction.


Microlepidogaster discus, a new species of Hypoptopomatinae (Siluriformes: Loricariidae) from the rio Jequitinhonha basin, southeastern Brazil.
A new Hypoptopomatinae is described from the upper rio Jequitinhonha basin. The new species is distinguished from all congeners by having the ventral laminar expansions of cleithrum and coracoid bearing only 1-30 odontodes, restricted to the area near the pectoral-fin insertion; the anterior plates of mid-ventral lateral series reduced, not ventrally elongated, the first or the second markedly round; pectoral-fin unbranched ray distinctly divided distally, with 7-15 ossified segments, reaching almost half of its length; and accessory process on first epibranchial present. Microlepidogaster discus is further distinguished from its congeners by several other osteological features. The problems concerning the taxonomy of Microlepidogaster, as well as the pattern of distribution of the genus, are discussed.


Turkish Validity and Reliability Study of Psychosocial Assessment Tool Oncology Version.
Children diagnosed with cancer and their families may not be able to overcome the effects of cancer diagnosis. Because there may be risk factors for some psychosocial problems. It is important to evaluate these risk factors and plan early interventions for risks such as depression, anxiety, suicide, and cessation of treatment. Psychosocial Assessment Tool was adapted to many languages. This study aimed to conduct the Turkish language adaptation of the Psychosocial Assessment Tool oncology version in families of 2- to 18-year-old children with newly diagnosed cancer. The steps used in the cultural adaptation of the scale were content validity, construct validity, criterion-related validity, surface validity, standard deviation and item analysis, internal consistency analysis, and test-retest reliability. We used the following scales for receiver operating characteristic analysis and correlations: the Pediatric Quality of Life Scale, the Hospital Anxiety Depression Scale, and Negative Problem Orientation Questionnaire. The reliability of Psychosocial Assessment Tool was strong [Kuder Richardson-20: 0.84], and the KR-20 values of the subdimensions ranged from 0.524 to 0.798. The Psychosocial Assessment Tool was moderately correlated with Pediatric Quality of Life Scale and Hospital Anxiety Depression Scale. Most families (49.4%) were classified in medium-risk category (targeted), 34.9% in low-risk category (universal), and 15.7% in high-risk category (clinical). The study confirms that the Turkish version of the Psychosocial Assessment Tool is a valid and reliable scale and could be used in pediatric oncology units for psychosocial evaluation and interventions in families of children with cancer.


Identification of human papillomavirus in esophageal squamous papillomas.
To investigate the presence of human papillomavirus (HPV) in esophageal squamous papilloma (ESP) and determine p16, p53 and Ki67 expression in a Mexican cohort. Nineteen cases diagnosed as ESP, corresponding to 18 patients were reviewed; nineteen cases of normal esophageal mucosa were used as negative controls. HPV detection was performed by amplified chromogenic in situ hybridization (ACISH) using a wide spectrum-cocktail probe and PCR. The average age at presentation was 46.3 years (range 28-72 years). Patients included four (22.22%) males and 14 (77.77%) females. The most frequent location was upper third (11 cases), followed by middle third (3 cases) and unknown site (5 cases). Immunohistochemistry (IHC) revealed basal and focal p53 expression in 17 cases (89%); p16 was expressed in eight cases (42.10%) and the Ki67 index ranged from 10% to 30%. HPV was detected in 14 out of 16 cases (87.5%) by ACISH: Twelve showed diffuse nuclear patterns and two showed granular patterns. HPV DNA was identified by PCR in 12 out of 14 cases (85.7%). Low-risk HPV types were detected in the most of the cases. This study provides identification of HPV infection in almost 80% of ESP using either ACISH or PCR; overall, all of these lesions show low expression of cell-cycle markers. We suggest ACISH as an alternative diagnostic tool for HPV detection in ESP.


A pilot randomised trial of catheter-directed thrombolysis or standard anticoagulation for patients with intermediate-high risk acute pulmonary embolism.
Intermediate-high risk acute pulmonary embolism (PE) remains associated with substantial mortality despite anticoagulation therapy. The aim of this randomised pilot study was to compare catheter-directed thrombolysis to standard anticoagulation therapy. Intermediate-high risk acute PE patients were admitted to a tertiary care centre (November 2019 to April 2021) and randomised in a 1:1 ratio to catheter-directed thrombolysis (CDT) or standard anticoagulation. Two catheters were used for the infusion of alteplase (1 mg/hr/catheter; total dose 20 mg) in the CDT group. The primary efficacy endpoint targeted improvement of right ventricular (RV) function, a decrease in pulmonary pressure, and a reduction of thrombus burden. Twenty-three patients were included (12 in the CDT group and 11 in the standard care group). The primary efficacy endpoint was achieved more frequently in the CDT group than in the standard care group (7 of 12 patients vs 1 of 11 patients, p=0.0004). An RV/left ventricular ratio reduction ≥25% (evident on computed tomography angiography) was achieved in 7 of 12 patients in the CDT group vs 2 of 11 patients in the standard care group (p=0.03). A systolic pulmonary artery pressure decrease of ≥30% or normotension at 24 hrs after randomisation was present in 10 of 12 patients in the CDT group vs 2 of 11 patients in the standard care group (p=0.001). There was no intracranial or life-threatening bleeding (type 5 or 3c bleeding, according to the Bleeding Academic Research Consortium classification). CDT for intermediate-high risk acute PE appears to be safe and effective. Further research is warranted to assess clinical endpoints.


Transductive Zero-Shot Learning With a Self-Training Dictionary Approach.
As an important and challenging problem in computer vision, zero-shot learning (ZSL) aims at automatically recognizing the instances from unseen object classes without training data. To address this problem, ZSL is usually carried out in the following two aspects: 1) capturing the domain distribution connections between seen classes data and unseen classes data and 2) modeling the semantic interactions between the image feature space and the label embedding space. Motivated by these observations, we propose a bidirectional mapping-based semantic relationship modeling scheme that seeks for cross-modal knowledge transfer by simultaneously projecting the image features and label embeddings into a common latent space. Namely, we have a bidirectional connection relationship that takes place from the image feature space to the latent space as well as from the label embedding space to the latent space. To deal with the domain shift problem, we further present a transductive learning approach that formulates the class prediction problem in an iterative refining process, where the object classification capacity is progressively reinforced through bootstrapping-based model updating over highly reliable instances. Experimental results on four benchmark datasets (animal with attribute, Caltech-UCSD Bird2011, aPascal-aYahoo, and SUN) demonstrate the effectiveness of the proposed approach against the state-of-the-art approaches.


Age-related scaling for aerosol and vapor deposition in the upper airways of humans.
Several theoretical models have been proposed in recent years to calculate age-related respiratory tract deposition of aerosols in humans. A critical factor in these models is the assumption about deposition in the airways above the trachea. In those models in which upper airway deposition is considered, a scaling factor, related to another airway dimension (usually tracheal diameter), is used to calculate an upper-airway dimension which determines inertial deposition. The paper reviews present knowledge of upper airway dimensions and airflow and proposed an age-related relationship for the critical nasal dimension, the nasal orifice. Relative inertial parameters for breathing at rest are obtained as a function of age from 1 mo to adulthood and compared to previous assumptions. The use of dimensionless parameters for predicting age-related deposition of diffusing particles and vapors is considered, and suggested experimental studies are proposed to supply needed information for age-related dosimetry of the entire respiratory tract.


Language-Specific Dual-Task Effects After Stroke: A Systematic Review.
The dual-task paradigm has been frequently used to examine stroke-related deficits because it samples behavioral performance under conditions of distraction similar to functioning in real-life environments. This original systematic review synthesizes studies that examined dual-task effects involving spoken language production in adults affected by stroke, including transient ischemic attack (TIA) and poststroke aphasia. Five databases were searched (inception to March 2022) for eligible peer-reviewed articles. The 21 included studies reported a total of 561 stroke participants. Thirteen studies focused on single word production, for example, word fluency, and eight on discourse production, for example, storytelling. Most studies included participants who had suffered a major stroke. Six studies focused on aphasia, whereas no study focused on TIA. A meta-analysis was not appropriate because of the heterogeneity of outcome measures. Some single word production studies found dual-task language effects whereas others did not. This finding was compounded by the lack of appropriate control participants. Most single word and discourse studies utilized motoric tasks in the dual-task condition. Our certainty (or confidence) assessment was based on a methodological appraisal of each study and information about reliability/fidelity. As 10 of the 21 studies included appropriate control groups and limited reliability/fidelity information, the certainty of the findings may be described as weak. Language-specific dual-task costs were identified in single word studies, especially those that focused on aphasia as well as half of the nonaphasia studies. Unlike single word studies, nearly all studies of discourse showed dual-task decrements on at least some variables. https://doi.org/10.23641/asha.23605311.


Polyclonal antibodies as probes to distinguish between tight and loose couple 50S ribosomes of Escherichia coli.
Antibody has been raised in rabbit against L7/L12 protein of E. coli 50S ribosomes and purified, finally through affinity column. A sensitive assay method using ELISA technique has also been standardised. LC 50S ribosomes react more with the antibody than TC 50S ribosomes. This supports the earlier physical data [Burma D P, Srivastava A K, Srivastava S, Tewari D S, Dash D & Sengupta S K, (1984), Biochem Biophys Res Commun, 124, 970] indicating that L7/L12 stalk region is protruded in medium in LC ribosomes and folded towards the body in TC ribosomes.


Detection and decay rates of prey and prey symbionts in the gut of a predator through metagenomics.
DNA methods are useful to identify ingested prey items from the gut of predators, but reliable detection is hampered by low amounts of degraded DNA. PCR-based methods can retrieve minute amounts of starting material but suffer from amplification biases and cross-reactions with the predator and related species genomes. Here, we use PCR-free direct shotgun sequencing of total DNA isolated from the gut of the harlequin ladybird Harmonia axyridis at five time points after feeding on a single pea aphid Acyrthosiphon pisum. Sequence reads were matched to three reference databases: Insecta mitogenomes of 587 species, including H. axyridis sequenced here; A. pisum nuclear genome scaffolds; and scaffolds and complete genomes of 13 potential bacterial symbionts. Immediately after feeding, multicopy mtDNA of A. pisum was detected in tens of reads, while hundreds of matches to nuclear scaffolds were detected. Aphid nuclear DNA and mtDNA decayed at similar rates (0.281 and 0.11 h(-1) respectively), and the detectability periods were 32.7 and 23.1 h. Metagenomic sequencing also revealed thousands of reads of the obligate Buchnera aphidicola and facultative Regiella insecticola aphid symbionts, which showed exponential decay rates significantly faster than aphid DNA (0.694 and 0.80 h(-1) , respectively). However, the facultative aphid symbionts Hamiltonella defensa, Arsenophonus spp. and Serratia symbiotica showed an unexpected temporary increase in population size by 1-2 orders of magnitude in the predator guts before declining. Metagenomics is a powerful tool that can reveal complex relationships and the dynamics of interactions among predators, prey and their symbionts.


[Neuroendocrine effects on one's awareness of the action of the heart].
Heart rate under mental and physical stress and the incidence of cardiac arrhythmias (by 24-hour ECG monitoring) as well as their awareness of these two types of cardiac activity were analysed in 11 insulin-dependent diabetics (manifestation before the age of 40 years) with autonomic neuropathy, five patients with hyperthyroidism and 13 healthy subjects. In hyperthyroidism the heart rate was increased as was the incidence of ventricular premature systoles; spontaneous variations and exercise-induced adaptation in heart rate were preserved. In diabetic neuropathy spontaneous heart-rate variations were absent, but adaptation reactions to mental and physical stress were normal. Patients of both groups had an impaired awareness of normal and arrhythmic cardiac activity. Lack of vagal inhibition in diabetic neuropathy and noradrenergic activation in hyperthyroidism may be the respective causative factors in the described disorders of regulation and awareness.


Centrosome-related genes, genetic variation, and risk of breast cancer.
Centrosome amplification has been detected in premalignant lesions and in situ tumors in the breast and in over 70% of invasive breast tumors, and has been associated with aneuploidy and tumor development. Based on these observations, the contribution of commonly inherited genetic variation in candidate genes related to centrosome structure and function to breast cancer risk was evaluated in an association study. Seven-hundred and 82 single nucleotide polymorphisms (SNPs) from 101 centrosomal genes were analyzed in 798 breast cancer cases and 843 controls from the Mayo Clinic Breast Cancer Study to assess the association between these SNPs (both individually and combined) and risk of breast cancer in this population. Eleven SNPs out of 782 from six genes displayed associations with breast cancer risk (P < 0.01). Haplotypes in five genes also displayed significant associations with risk. A two SNP combination of rs10145182 in NIN and rs2134808 in the TUBG1 locus (P-interaction = 0.00001), suggested SNPs in mediators of microtubule nucleation from the centrosome contribute to breast cancer. Evaluation of the simultaneous significance of all SNPs in the centrosome pathway suggested that the centrosome pathway is highly enriched (P = 4.76 × 10(-50)) for SNPs that are associated with breast cancer risk. Collections of weakly associated genetic variants in the centrosome pathway, rather than individual highly significantly associated SNPs, may account for a putative role for the centrosome pathway in predisposition to breast cancer.


Risk factors for the development of acute lung injury in patients with infectious pneumonia.
Although pneumonia has been identified as the single most common risk factor for acute lung injury (ALI), we have a limited knowledge as to why ALI develops in some patients with pneumonia and not in others. The objective of this study was to determine frequency, risk factors, and outcome of ALI in patients with infectious pneumonia. A retrospective cohort study of adult patients with microbiologically positive pneumonia, hospitalized at two Mayo Clinic Rochester hospitals between January 1, 2005, and December 31, 2007. In a subsequent nested case-control analysis, we evaluated the differences in prehospital and intrahospital exposures between patients with and without ALI/acute respiratory distress syndrome (ARDS) matched by specific pathogen, isolation site, gender, and closest age in a 1:1 manner. The study included 596 patients; 365 (61.2%) were men. The median age was 65 (IQR, 53 to 75) years. In total, 171 patients (28.7%) were diagnosed with ALI. The occurrence of ALI was less frequent in bacterial (n = 99 of 412, 24%) compared with viral (n = 19 of 55, 35%), fungal (n = 39 of 95, 41%), and mixed isolates pneumonias (n = 14 of 34, 41%; P = 0.002). After adjusting for baseline severity of illness and comorbidities, patients in whom ALI developed had a markedly increased risk of hospital death (ORadj 9.7; 95% CI, 6.0 to 15.9). In a nested case-control study, presence of shock (OR, 8.9; 95% CI, 2.8 to 45.9), inappropriate initial antimicrobial treatment (OR, 3.2; 95% CI, 1.3 to 8.5), and transfusions (OR, 4.8; 95% CI, 1.5 to 19.6) independently predicted ALI development. The development of ALI among patients hospitalized with infectious pneumonia varied among pulmonary pathogens and was associated with increased mortality. Inappropriate initial antimicrobial treatment and transfusion predict the development of ALI independent of pathogen.


Men's experiences of grandfatherhood: a welcome surprise.
The present-day involvement of men in many facets of childrearing stands in contrast to previous eras when men accepted that the major task of fathering was to provide a secure income to support the family. This imperative often required long hours away from the family. However, when men whose contact with children has been limited due to work and cultural constraints retire, their newly acquired lifestyle may bring fresh opportunities for involvement with grandchildren. An important question therefore concerns the impact of caring for young children on men's perceptions of their role as grandfathers. This interview study explores the experiences of 19 Australian grandfathers. The analysis found themes that relate to relationships and change, as well as themes concerning core beliefs and existential questions. The findings demonstrate the potential for insight into family relationships and personal growth in older age when studying the topic of grandparenting and caring from the male perspective.


Degenerate Elastic Networks.
We minimize a linear combination of the length and the L 2 -norm of the curvature among networks in R d belonging to a given class determined by the number of curves, the order of the junctions, and the angles between curves at the junctions. Since this class lacks compactness, we characterize the set of limits of sequences of networks bounded in energy, providing an explicit representation of the relaxed problem. This is expressed in terms of the new notion of degenerate elastic networks that, rather surprisingly, involves only the properties of the given class, without reference to the curvature. In the case of d = 2 we also give an equivalent description of degenerate elastic networks by means of a combinatorial definition easy to validate by a finite algorithm. Moreover we provide examples, counterexamples, and additional results that motivate our study and show the sharpness of our characterization.


Physicochemical Characterization and Drug Release Properties of Methyl-Substituted Silica Xerogels Made Using Sol-Gel Process.
In this work, a multi-analytical approach involving nitrogen porosimetry, small angle neutron and X-ray scattering, Fourier transform infrared (FTIR) and nuclear magnetic resonance (NMR) spectroscopies, X-ray diffraction, thermal analysis and electron microscopy was applied to organically modified silica-based xerogels obtained through the sol-gel process. Starting from a tetraethoxysilane (TEOS) precursor, methyltriethoxysilane (MTES) was added to the reaction mixture at two different pH values (2.0 and 4.5) producing hybrid xerogels with different TEOS/MTES molar ratios. Significant differences in the structure were revealed in terms of the chemical composition of the silica network, hydrophilic/hydrophobic profile, particle dimension, pore shape/size and surface characteristics. The combined use of structural characterization methods allowed us to reveal a relation between the cavity dimensions, the synthesis pH value and the grade of methyl substitution. The effect of the structural properties on the controlled Captopril release efficiency has also been tested. This knowledge facilitates tailoring the pore network for specific usage in biological/medical applications. Knowledge on structural aspects, as reported in this work, represents a key starting point for the production of high-performance silica-based hybrid materials showing enhanced efficacy compared to bare silica prepared using only TEOS.


Genome-wide association studies of cancer.
Knowledge of the inherited risk for cancer is an important component of preventive oncology. In addition to well-established syndromes of cancer predisposition, much remains to be discovered about the genetic variation underlying susceptibility to common malignancies. Increased knowledge about the human genome and advances in genotyping technology have made possible genome-wide association studies (GWAS) of human diseases. These studies have identified many important regions of genetic variation associated with an increased risk for human traits and diseases including cancer. Understanding the principles, major findings, and limitations of GWAS is becoming increasingly important for oncologists as dissemination of genomic risk tests directly to consumers is already occurring through commercial companies. GWAS have contributed to our understanding of the genetic basis of cancer and will shed light on biologic pathways and possible new strategies for targeted prevention. To date, however, the clinical utility of GWAS-derived risk markers remains limited.


A Macroscopic Quantum Three-Box Paradox: Finding Consistency with Weak Macroscopic Realism.
The quantum three-box paradox considers a ball prepared in a superposition of being in any one of three boxes. Bob makes measurements by opening either box 1 or box 2. After performing some unitary operations (shuffling), Alice can infer with certainty that the ball was detected by Bob, regardless of which box he opened, if she detects the ball after opening box 3. The paradox is that the ball would have been found with certainty by Bob in either box if that box had been opened. Resolutions of the paradox include that Bob's measurement cannot be made non-invasively or else that realism cannot be assumed at the quantum level. Here, we strengthen the case for the former argument by constructing macroscopic versions of the paradox. Macroscopic realism implies that the ball is in one of the boxes prior to Bob or Alice opening any boxes. We demonstrate the consistency of the paradox with macroscopic realism, if carefully defined (as weak macroscopic realism, wMR) to apply to the system at the times prior to Alice or Bob opening any boxes but after the unitary operations associated with preparation or shuffling. By solving for the dynamics of the unitary operations and comparing with mixed states, we demonstrate agreement between the predictions of wMR and quantum mechanics: the paradox only manifests if Alice's shuffling combines both local operations (on box 3) and nonlocal operations, on the other boxes. Following previous work, the macroscopic paradox is shown to correspond to a violation of a Leggett-Garg inequality, which implies failure of non-invasive measurability if wMR holds.


Change-point analysis of neuron spike train data.
In many medical experiments, data are collected across time, over a number of similar trials, or over a number of experimental units. As is the case of neuron spike train studies, these data may be in the form of counts of events per unit of time. These counts may be correlated within each trial. It is often of interest to know if the introduction of an intervention, such as the application of a stimulus, affects the distribution of the counts over the course of the experiment. In such investigations, each trial generates a sequence of data that may or may not contain a change in distribution at some point in time. Each sequence of integer counts can be viewed as arising from a Poisson process and are therefore independently distributed or as an integer-valued time series that allows for correlations between these counts. The main aim of this paper is to show how the ensemble of sample paths may be used to make inference about the distribution of the instantaneous times of change in a given population. This will be accomplished using a Bayesian hierarchical model for these change-points in time. A bonus of these models is they also allow for inference about the probability of a change in each unit and the magnitude of the effects, if any. The use of such change-point models on integer-valued time series is illustrated on neuron spike train data, although the methods can be applied to other situations where integer-valued processes arise.


50,000 years of ice and seals: Impacts of the Last Glacial Maximum on Antarctic fur seals.
Ice is one of the most important drivers of population dynamics in polar organisms, influencing the locations, sizes, and connectivity of populations. Antarctic fur seals, Arctocephalus gazella, are particularly interesting in this regard, as they are concomitantly reliant on both ice-associated prey and ice-free coastal breeding areas. We reconstructed the history of this species through the Last Glacial Maximum (LGM) using genomic sequence data from seals across their range. Population size trends and divergence events were investigated using continuous-time size estimation analysis and divergence time estimation models. The combined results indicated that a panmictic population present prior to the LGM split into two small refugial populations during peak ice extent. Following ice decline, the western refugial population founded colonies at the South Shetlands, South Georgia, and Bouvetøya, while the eastern refugial population founded the colony on Iles Kerguelen. Postglacial population divergence times closely match geological estimates of when these coastal breeding areas became ice free. Given the predictions regarding continued future warming in polar oceans, these responses of Antarctic fur seals to past climate variation suggest it may be worthwhile giving conservation consideration to potential future breeding locations, such as areas further south along the Antarctic Peninsula, in addition to present colony areas.


Discovery and validation of new potential biomarkers for early detection of colon cancer.
Accurate detection of characteristic proteins secreted by colon cancer tumor cells in biological fluids could serve as a biomarker for the disease. The aim of the present study was to identify and validate new serum biomarkers and demonstrate their potential usefulness for early diagnosis of colon cancer. The study was organized in three sequential phases: 1) biomarker discovery, 2) technical and biological validation, and 3) proof of concept to test the potential clinical use of selected biomarkers. A prioritized subset of the differentially-expressed genes between tissue types (50 colon mucosa from cancer-free individuals and 100 normal-tumor pairs from colon cancer patients) was validated and further tested in a series of serum samples from 80 colon cancer cases, 23 patients with adenoma and 77 cancer-free controls. In the discovery phase, 505 unique candidate biomarkers were identified, with highly significant results and high capacity to discriminate between the different tissue types. After a subsequent prioritization, all tested genes (N = 23) were successfully validated in tissue, and one of them, COL10A1, showed relevant differences in serum protein levels between controls, patients with adenoma (p = 0.0083) and colon cancer cases (p = 3.2e-6). We present a sequential process for the identification and further validation of biomarkers for early detection of colon cancer that identifies COL10A1 protein levels in serum as a potential diagnostic candidate to detect both adenoma lesions and tumor. The use of a cheap serum test for colon cancer screening should improve its participation rates and contribute to decrease the burden of this disease.


Pulmonary involvement in Fabry disease: overview and perspectives.
Fabry disease (FD) is an X-linked lysosomal storage disorder caused by deficiency of alpha-galactosidase A, which leads to storage of sphingolipids in virtually all human cells and consequently to organ dysfunction. Pulmonary involvement is still debated. But, obstructive lung disease is up to ten times more prevalent in patients with FD compared to general public. Also, an accelerated decline in forced expiratory volume in one second (FEV1) over time was observed in these patients. Lysosomal storage of glycosphingolipids is considered leading to small airway disease via hyperplasia of the bronchiolar smooth muscle cells. Larger airways may become involved with ongoing disease process. There is no evidence for involvement of the lung interstitium in FD. The effect of enzyme replacement therapy on respiratory involvement remains to be determined in large, prospective controlled trials.


Is Apixaban Safe and Effective for Venous Thromboembolism Prophylaxis After Primary Total Hip and Total Knee Arthroplasties?
Venous thromboembolism (VTE) is a serious complication of total hip arthroplasty (THA) and total knee arthroplasty (TKA). Apixaban is approved for VTE prophylaxis. This study seeks to ascertain the risk of VTE and bleeding complications in patients undergoing primary THA and TKA receiving apixaban for postoperative VTE prophylaxis for one of the following indications: high risk for VTE, previously on apixaban, and contraindication to the use of aspirin. This is a retrospective cohort study of patients who underwent primary THA or TKA over a 17-month period and were prescribed apixaban for thromboprophylaxis postoperatively. 230 patients were included in the study, 110 TKA and 120 THA. The primary reasons for high-risk VTE status included personal and family history of VTE, and 13% were taking apixaban preoperatively for atrial fibrillation. 1 patient (0.43%) who underwent TKA had a DVT with PE. 2.6% of patients had wound complications requiring operative treatment, and 0.87% of THA patients underwent revision arthroplasty. The use of apixaban for VTE prophylaxis after primary THA and TKA in patients at high risk for VTE, in patients previously on apixaban, and in patients with a contraindication to the use of aspirin is associated with a low risk of VTE and bleeding complications.


ASCO update: lung cancer.
In the past few years there have been major changes in the treatment landscape in oncology; lung cancer is affected by those changes like almost no other solid tumor. The rise of further second- and third-line tyrosine kinase inhibitors offers sequential therapy for patients with mutated non-small-cell lung cancer. Immunotherapy has found its way into clinical routine and presents us with new challenges in managing side effects, evaluating treatment response and deciding on how long we treat our patients. The treatment algorithm of lung cancer has changed in the last month and further practice-changing trials are coming up, so treating lung cancer patients shows nowadays a more challenging perspective with the possibility of subsequently applied individual therapies. This article provides a brief overview of the highlights presented at the ASCO (American Society of Clinical Oncology) annual meeting this year in Chicago.


Analytical methods for the determination of bisphenol A in food.
Food constitutes the primary route for human exposure to bisphenol A (BPA), one of the highest volume chemicals produced worldwide. The estrogenic properties of BPA, its wide dispersive use and the recent extensive literature describing low-dose BPA effects in animals, have raised concerns about its possible adverse effects on human health. A reliable health risk assessment of BPA relies basically on its unambiguous identification and accurate quantification in food, and the aim of the present review is to give an overview of the analytical methods reported so far for the determination of BPA in these matrices. Emphasis is placed on the main strategies developed for sample treatment, which usually consists of several laborious and time-consuming steps in order to achieve the required sensitivity and selectivity. Separation, identification and quantitation of BPA is today reliably made with mass spectrometric methods, namely liquid chromatography-mass spectrometry (LC-MS) and gas chromatography-mass spectrometry (GC-MS), and thus main attention is devoted to these techniques, but other methods using LC coupled to fluorescence or electrochemical detection, as well as immunochemical methods are also covered. Recent and expected future developments are discussed.


Development of real-time reverse transcriptase PCR assays to detect and serotype dengue viruses.
Serotyping dengue virus (DENV) from suspect human specimens is crucial for developing sound epidemiological control measurements early in the transmission season and for effective patient management. We modified DENV consensus D1 (mD1) and serotype-specific TS2 (mTS2) and redesigned serotype-specific TS1 (rTS1) and TS4 (rTS4) as described previously in the conventional capsid and premembrane gene (C-prM) protocol (R. S. Lanciotti, C. H. Calisher, D. J. Gubler, G.-J. Chang, A. V. Vorndam, J. Clin. Microbiol. 30:545-551, 1992). In addition, we designed two new sets of amplimers and probes, located at nonstructural protein 5 (NS5) and the 3' noncoding region (3'NC) of DENV. The NS5 protocol utilizes two flaviviral consensus outer amplimers (mFU1 and CFD2) and four dengue virus serotype-specific TaqMan fluorogenic probes. The 3'NC protocol uses two DENV consensus amplimers, DC10418 and CDC10564. The conventional gel-based, heminested detection method was adapted for the C-prM protocol for detecting and serotyping dengue viruses. In addition, we developed the real-time SYBR green I and postamplification melting temperature curve analysis for the mD1/TS and 3'NC protocols using identical amplification conditions. The NS5 amplimer/probe set was formulated as a one-tube, multiplex, real-time reverse transcriptase PCR for serotype identification. Three sets of amplimers and probes were verified for their specificity in tests with yellow fever, Japanese encephalitis, St. Louis encephalitis, and West Nile viruses; optimized against 109 DENV strains; and validated for detection of the virus in sera from two different panels of acute-phase human dengue serum specimens and one panel of virus isolates from dengue patients' serum specimens. Clinical evaluation by two separate laboratories indicated that the C-prM was more sensitive (100%) than the NS5 (91%) or the 3'NC (91%) protocol.


Small intestinal motility and transit in disease.
Disorders of small intestinal motility and transit are becoming increasingly recognized partly as a result of a greater awareness of their existence and partly because suitable diagnostic methods are more widely available. Usually, the neuropathic and myopathic forms can be separated, and gut disease secondary to a generalized neuromuscular disorder can be identified by the clinician. The availability of better non-invasive methods for the diagnosis of disorders of motility and transit would greatly facilitate their management. Treatment must include the restoration and maintenance of nutrition, attempts to improve intestinal motor function and resection of any segments of localized disease. Regrettably, all such measures are ineffective in the severest cases. In the future, a greater understanding of the enteric neural control of the smooth muscle and an ability to manipulate it with novel, specific drugs or peptidergic receptor agonists and antagonists, or electrical pacing, may lead to more effective therapies.


[Record linkage with cryptographic identification data in a population-based cancer registry. Development, implementation and error rates].
Population-based cancer registries depend on the completeness of their case notification. At present, restrictive legal regulations in Germany requesting written informed consent of cancer patients hinder some health professionals, for example pathologists, from reporting to cancer registries. New cryptographic methods may be used to obtain record linkage based on anonymised data. To assess the feasibility of a record linkage in the Münster Cancer Registry (EKR-MS) using cryptographic methods as compared to traditional methods based on personal identifiers. We generated so called control numbers--a sequence of 23 alphanumeric signs--for 19 name related characteristics employing consecutively two different chiffrations. Record linkage was carried out using a semi-automatic computer program (AUTOMATCH) that generates probabilities of identity for pairs of case notifiation based on the information of control numbers only. Probabilities exceeding present limits lead to automatic decisions whereas the remaining linkage has to be decided manually. Plausibility-based rules helped to considerably reduce the amount of manual decisions. We compared traditional and cryptographic record linkage for all new reports received in the EKR-MS during 1998 (n = 27,262) against the background of n = 101,880 known cases in the registry data base. Setting traditional, text-based record linkage results to be the reference the cryptographic method resulted in a synonomous error (false creation of a new case) of almost 2% and a homonymous error (false link to a known case) of less than 0.5%. Cryptographic methods may be feasible procedures of record linkage in cancer registries. The size of the database of the EKR-MS prohibits extrapolation of findings to smaller registries. The error rates resulting in slight overestimation of disease rates that may seem acceptable. It should be noted though that cryptographic data are in many situations prohibitive for further epidemiologic research limiting the usefulness of the method exclusively to cancer registration under the given legal coercion.


Autopsy of a failed trial part 1: A qualitative investigation of clinician's views on and experiences of the implementation of the DAISIES trial in UK-based intensive eating disorder services.
The DAISIES trial, comparing inpatient and stepped-care day patient treatment for adults with severe anorexia nervosa was prematurely terminated in March 2022 due to poor recruitment. This qualitative study seeks to understand the difficulties faced during the trial by investigating stakeholders' views on and experiences of its implementation. Semi-structured interview and focus group transcripts, and trial management and oversight group meeting minutes from May 2020-June 2022 were analysed using thematic analysis. Participants were 47 clinicians and co-investigators involved with the DAISIES trial. The Non-Adoption, Abandonment, Scale-up, Spread, and Sustainability (NASSS) framework was applied to the interpretive themes to classify barriers and facilitators to implementation. Five themes were identified: incompatible participation interests; changing standard practice; concerns around clinical management; systemic capacity and capability issues; and Covid-19 disrupting implementation. Applying the NASSS framework indicated the greatest implementation challenges to arise with the adopters (e.g. patients, clinicians), the organisational systems (e.g. service capacity), and the wider socio-political context (e.g. Covid-19 closing services). Our findings emphasise the top-down impact of systemic-level research implementation challenges. The impact of the Covid-19 pandemic accentuated pre-existing organisational barriers to trial implementation within intensive eating disorder services, further limiting the capacity for research.


Effects of HOXB4 overexpression on ex vivo expansion and immortalization of hematopoietic cells from different species.
Overexpression of the human HOXB4 has been shown to induce the expansion and self-renewal of murine hematopoietic stem cells. In preparation for clinical studies, we wished to investigate the effects of HOXB4 on cells from other species, in particular preclinical large animals such as dogs and nonhuman primates. Thus, we transduced CD34(+) cells from nonhuman primates, dogs, and humans with a HOXB4-expressing gammaretroviral vector and a yellow fluorescent protein-expressing control vector. Compared with the control vector, HOXB4 overexpression resulted in a much larger increase in colony-forming cells in dog cells (28-fold) compared with human peripheral blood, human cord blood, and baboon cells (two-, four-, and fivefold, respectively). Furthermore, we found that HOXB4 overexpression resulted in immortalization with sustained growth (>12 months) of primitive hematopoietic cells from mice and dogs but not from monkeys and humans. This difference correlated with increased levels of retrovirally overexpressed HOXB4 in dog and mouse cells compared with human and nonhuman primate cells. The immortalized cells did not show any evidence of insertional mutagenesis or chromosomal abnormalities. Competitive congenic transplantation experiments showed that HOXB4-expanded mouse cells engrafted well after 1 or 3 months of expansion, and no leukemia was observed in mice. Our findings suggest that the growth promoting effects of HOXB4 are critically dependent on HOXB4 expression levels and that this can result in important species-specific differences in potency. Disclosure of potential conflicts of interest is found at the end of this article.


Observation of extremely heterogeneous electroporative molecular uptake by Saccharomyces cerevisiae which changes with electric field pulse amplitude.
Molecular uptake of a charged fluorescent molecule (calcein; 623 Da, z = -4) was quantitatively determined at the single cell level using flow cytometry. Dilutely suspended cells were exposed to one exponential pulse (tau p approximately 300 microseconds) for different field strength values. For an asymmetric cell such as the yeast Saccharomyces cerevisiae a significant variation in the number of molecules taken up by individual cells was expected for physical reasons. By carrying out several thousand individual cell measurements for each pulse condition, we found that the number of molecules per cell varies significantly within the cell population, and that this population distribution changes markedly as the field strength is varied. Surprisingly, in spite of significant changes in this distribution with field strength, the average uptake per cell reaches a non-equilibrium plateau for which the uptake per cell is much smaller than the product of the mean cell volume and the supplied extracellular concentration. These observations of different field-dependent cell population distributions of uptake support the hypotheses that (1) electroporation is a transmembrane voltage-responsive phenomenon, so that cells of different sizes, shapes and orientation, respond differently to even a spatially uniform applied field, (2) population average measurements of electroporation behavior can be incomplete and misleading, and (3) transport of small charged molecules is due to electrophoresis through the pores of a dynamically changing pore population.


Amino acids biosynthesis and nitrogen assimilation pathways: a great genomic deletion during eukaryotes evolution.
Besides being building blocks for proteins, amino acids are also key metabolic intermediates in living cells. Surprisingly a variety of organisms are incapable of synthesizing some of them, thus named Essential Amino Acids (EAAs). How certain ancestral organisms successfully competed for survival after losing key genes involved in amino acids anabolism remains an open question. Comparative genomics searches on current protein databases including sequences from both complete and incomplete genomes among diverse taxonomic groups help us to understand amino acids auxotrophy distribution. Here, we applied a methodology based on clustering of homologous genes to seed sequences from autotrophic organisms Saccharomyces cerevisiae (yeast) and Arabidopsis thaliana (plant). Thus we depict evidences of presence/absence of EAA biosynthetic and nitrogen assimilation enzymes at phyla level. Results show broad loss of the phenotype of EAAs biosynthesis in several groups of eukaryotes, followed by multiple secondary gene losses. A subsequent inability for nitrogen assimilation is observed in derived metazoans. A Great Deletion model is proposed here as a broad phenomenon generating the phenotype of amino acids essentiality followed, in metazoans, by organic nitrogen dependency. This phenomenon is probably associated to a relaxed selective pressure conferred by heterotrophy and, taking advantage of available homologous clustering tools, a complete and updated picture of it is provided.


Hispanic Patients with Primary Biliary Cholangitis Have Decreased Access to Care Compared to Non-Hispanics.
Background and Aims: Hispanic patients with primary biliary cholangitis (PBC) have reduced rates of biochemical response to ursodeoxycholic acid (UDCA) and increased risk of disease progression compared to non-Hispanic patients. In this study, we sought to identify differences in demographics, comorbidities, environmental risk factors and socioeconomic status between Hispanic and non-Hispanic patients with PBC. Methods: In a case control study, we analyzed data from Hispanic (n=37 females and 1 male) and non-Hispanic (n=54 females and 4 males) patients with PBC seen at the University of Miami/Jackson Memorial Hospital from January 1998 through January 2013. Data were obtained by filling out a questionnaire either via phone call, mail, or e-mail. Odds ratios were calculated to measure the association between exposure and outcomes. Results: Baseline demographics, environmental risk factors and comorbidities were similar between Hispanic and non-Hispanic patients with PBC. Hispanic patients were less likely to be married and fewer Hispanics had education beyond high school level compared to non-Hispanics. Sixty four percent of Hispanic patients had a household income of less than $50000, compared to 19.5% of non-Hispanics. Fewer Hispanic patients with PBC had health insurance coverage compared to non-Hispanics (86.5% vs. 98.1%; odds ratio: 0.1, 95% confidence interval: 0-0.9). Conclusions: Differences in disease severity and response to therapy observed in prior studies could not be explained by environmental exposures. In addition to genetic variation, socioeconomic discrepancies (access to care) may further explain these differences.


Surveillance study for creating the national clinical database related to ECG-gated myocardial perfusion SPECT of ischemic heart disease: J-ACCESS study design.
ECG-gated myocardial perfusion SPECT is widely applied to diagnose ischemic heart disease, and such findings are useful to predict patient prognosis. However, Japan does not have a database that correlates SPECT image findings with the prognosis of patients who have ischemic heart disease. A large-scale clinical study involving 117 medical facilities throughout Japan was established to survey the clinical background and image findings of patients who have undergone ECG-gated stress perfusion SPECT. These patients were followed up for three years to investigate the occurrence of cardiac events. The 4,629 registered patients comprised 2,989 males (age 64.9 +/- 10.3 y, mean +/- SD) and 1,640 females (age 67.2 +/- 9.7 y). The most frequent complication was hypertension (54.5%), followed by hyperlipidemia (47.2%) and diabetes (29.4%). Percutaneous coronary intervention (PCI) or coronary artery bypass grafting (CABG) was conducted on 1,925 of the patients. SPECT examinations were ordered for further examination of chest pain (32.8%), periodic follow-up after coronary artery intervention (24.2%), screening for coronary artery disease (15.1%), follow-up of old myocardial infarction (14.9%), more detailed investigation of ECG or echocardiographic abnormalities (13.1%), etiological assessment of heart failure (1.6%), and further inspection for acute coronary syndrome (0.3%). The method of inducing stress was most often exercise loading at 68.8%, and infusion of either dipyridamole (14.6%) or adenosine triphosphate (ATP, 13.8%). The most frequently applied amount of 99mTc-tetrofosmin was an initial dose of 200 to 300 MBq combined with a second dose of 700 to 800 MBq (37.7%). The mean doses were 305 +/- 81 at the initial and 709 +/- 132 MBq at the second administration. A history of angina pectoris (41.2%) was the most frequent, followed by myocardial infarction (29.5%). During the two years of follow-up after registration, 46 of the 4,629 subjects have discontinued or dropped out, 134 have died, and 4,449 (97.8%) continue to undergo follow-up investigations. A complete report will be presented when the follow-up data for 3 years have been compiled and analyzed.


Self-consistent field method for open-shell systems within the density-matrix functional theory.
The unrestricted Hartree-Fock method is extended to correlation calculation within the density-matrix functional theory. The method is derived from an entropic cumulant functional for the correlation energy. The eigenvalue equations for the spin-orbitals are modified by the orbital occupation numbers. The Euler equation for the occupation numbers results in the Fermi-Dirac distribution, which is very efficient to update as soon as the orbital eigenvalue equations are solved. The method is demonstrated on the ground state of O 2 $$ {}_2 $$ .


Descending facilitation from the rostral ventromedial medulla maintains nerve injury-induced central sensitization.
Nerve injury can produce hypersensitivity to noxious and normally innocuous stimulation. Injury-induced central (i.e. spinal) sensitization is thought to arise from enhanced afferent input to the spinal cord and to be critical for expression of behavioral hypersensitivity. Descending facilitatory influences from the rostral ventromedial medulla have been suggested to also be critical for the maintenance, though not the initiation, of experimental neuropathic pain. The possibility that descending facilitation from the rostral ventromedial medulla is required for the maintenance of central sensitization was examined by determining whether ablation of mu-opioid receptor-expressing cells within the rostral ventromedial medulla prevented the enhanced expression of repetitive touch-evoked FOS within the spinal cord of animals with spinal nerve ligation injury as well as nerve injury-induced behavioral hypersensitivity. Rats received a single microinjection of vehicle, saporin, dermorphin or dermorphin-saporin into the rostral ventromedial medulla and 28 days later, underwent either sham or spinal nerve ligation procedures. Animals receiving rostral ventromedial medulla pretreatment with vehicle, dermorphin or saporin that were subjected to spinal nerve ligation demonstrated both thermal and tactile hypersensitivity, and showed significantly increased expression of touch-evoked FOS in the dorsal horn ipsilateral to nerve injury compared with sham-operated controls at days 3, 5 or 10 post-spinal nerve ligation. In contrast, nerve-injured animals pretreated with dermorphin-saporin showed enhanced behaviors and touch-evoked FOS expression in the spinal dorsal horn at day 3, but not days 5 and 10, post-spinal nerve ligation when compared with sham-operated controls. These results indicate the presence of nerve injury-induced behavioral hypersensitivity associated with nerve injury-induced central sensitization. Further, the results demonstrate the novel concept that once initiated, maintenance of nerve injury-induced central sensitization in the spinal dorsal horn requires descending pain facilitation mechanisms arising from the rostral ventromedial medulla.


From superior adaptation and function to brain dysfunction--the neglect of epigenetic factors.
With optimal pregnancy conditions (natural, enriched diet which includes fish) African (Digo) infants are 3-4 weeks ahead of European/American infants in sensorimotor terms at birth, and during the first year. Infants of semi-aquatic sea-gypsies swim before they walk, and have superior visual acuity compared with us. With adverse pregnancy behaviour (fear of fat, a trend to dieting), neglecting the need for brain fat to secure normal brain development and function, we run a risk of dysfunction--death. Sudden Infant Death Syndrome victims have depressed birth weight, lower levels of marine fat in brainstem than controls, and >80 suffer multiple hypoxic episodes prior to death. Depressed birth weight (more than 10% below mean) is seen in learning and behaviour disorders, and a trend towards weights of less than 3kg is increasing, which supports a rise in antenatal sub optimality. Given marine fat deficiency in pregnancy and infancy, neurons starved for fuel could delay myelination and maturation in the latest developed Frontal Lobes. The phylogenetic oldest Lateral Frontal Lobe System (feed-back mechanism etc.) derived from olfactory bulb-amygdala, which crosses in Anterior Commisure is probably spared, while the Medial Frontal Lobe System derived from Hippocampus-Cingulum and crosses in Corpus Callosum (delayed response task) is most likely affected. The rise in infantile autism (intact vision and hearing) with deficit in delayed response task only, could suggest a deficit in the Medial Frontal Lobe System. The human species is unique; 70% of total energy to the foetus goes to development of the brain, which mainly consists of marine fat. It undergoes pervasive regressive events, before birth, in infancy and at puberty. Minimal retraction of neuronal arborisation is advantageous. Attributable to adverse pregnancy childrearing practice, excessive retraction is likely prenatally and in infancy. Pubertal age affects the fundamental property of nervous tissue, excitability: excessive excitatory drive is seen in early, and a deficiency in late puberty. It is postulated that with adequate marine fat, there is probably no risk of psychopathology at the extremes, whereas a deficiency could lead to paroxysmal (subcortical) dysfunction in early puberty, and breakdown of cortical circuitry and cognitive dysfunctions in late puberty. The post-pubertal psychoses, schizophrenia and manic-depressive psychosis at the extremes of the pubertal age continuum, with contrasting excitability and biological treatment, are probably the result of continuous dietary deficiency, which has inactivated the expression of genes for myelin development and oligodendrocyte-related genes in their production of myelin. The beneficial effect of marine fat in both disorders, in other CNS disorders as well as in developmental dyslexia (DD) and ADHD among others, supports our usual diet is persistently deficient. We have neglected the similarity of our great brain to other mammals, and our marine heritage. Given the amount of marine fat needed to secure normal brain development and function is not known, nor the present dietary level, it seems unduly conjectural to postulate that a dietary deficiency in marine fat is causing brain dysfunction and death. However, all observations point in the same direction: our diet focusing on protein mainly, is deficient, the deficiency is most pronounced in maternal nutrition and in infancy.


A protein antigenically related to nuclear lamin B mediates the association of intermediate filaments with desmosomes.
Desmosomes are specialized domains of epithelial cell plasma membranes engaged in the anchoring of intermediate filaments (IF). So far, the desmosomal component(s) responsible for this binding has not been unambiguously identified. In the present work, we have examined bovine muzzle epidermis desmosomes for the presence of protein(s) structurally and functionally related to lamin B, the major receptor for IF in the nuclear envelope (Georgatos, S. D., and G. Blobel. 1987. J. Cell Biol. 105:105-115). By using polyclonal antibodies to lamin B in immunoblotting experiments, we find that a desmosomal protein of 140-kD shares epitope(s) with lamin B. Immunoelectron microscopic and urea extraction experiments show that this protein is a peripheral protein localized at the cytoplasmic side of the desmosomes (desmosomal plaques). Furthermore, this protein binds vimentin in an in vitro assay. Since this binding is inhibited by lamin B antibodies, the epitopes common to the 140-kD protein and to lamin B may be responsible for anchoring of intermediate filaments to desmosomes. These data suggest that lamin B-related proteins (see also Cartaud, A., J. C. Courvalin, M. A. Ludosky, and J. Cartaud. 1989. J. Cell Biol. 109:1745-1752) together with lamin B, provide cells with several nucleation sites, which can account for the multiplicity of IF organization in tissues.


Vitamin D supplementation dose needs to be higher in patients with inflammatory bowel disease: interventional study.
Vitamin D (vitD) is a substance with an immunomodulatory effect. Its insufficiency has negative impact also on inflammatory bowel disease (IBD) where it is often present. The recommended daily intake for general population is 600 UI/day (units/day). What is the necessary dose for IBD patients remains unknown. The aim of the study was to verify whether the 2,000 IU/day of vitD is sufficient for maintaining sustained levels in these patients. Patients with Crohns disease (CD) or ulcerative colitis (UC) were supplemented orally with 2,000 IU of cholecalciferol daily during winter time. The level of 25-hydroxyvitamin D (25OHD) was established at the beginning of substitution period (October to December) and in Month 4. Demographic data, Ca, P, parathormone levels, dose of vitD used, and patients compliance were observed. 108 patients with CD and UC (71/37) were analysed, out of them 51 females, average age 43.3 ± 16.2 years. The level of 25OHD increased from 60.2 ± 26.5 nmol/l to 68.1 ± 27.1 nmol/l (p < 0.001) during the period with the average substitution dose of vitD 1 858 ± 464 IU/day. 60.2 % of subjects complied with the recommended dosing of vitD. The dose of 1,820 IU vitD/day showed to be sufficient for maintaining sustained levels in the model. No changes of Ca, P serum levels occurred during observation period. Substitution doses of vitD recommended for general population are insufficient for IBD patients. A dose of up to 2,000/day, which is safe, is necessary to maintain normal levels of 25OHD. Noncompliance with the use of vitD is high.


Symptom-Based Predictive Model of COVID-19 Disease in Children.
Testing for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is neither always accessible nor easy to perform in children. We aimed to propose a machine learning model to assess the need for a SARS-CoV-2 test in children (<16 years old), depending on their clinical symptoms. Epidemiological and clinical data were obtained from the REDCap® registry. Overall, 4434 SARS-CoV-2 tests were performed in symptomatic children between 1 November 2020 and 31 March 2021, 784 were positive (17.68%). We pre-processed the data to be suitable for a machine learning (ML) algorithm, balancing the positive-negative rate and preparing subsets of data by age. We trained several models and chose those with the best performance for each subset. The use of ML demonstrated an AUROC of 0.65 to predict a COVID-19 diagnosis in children. The absence of high-grade fever was the major predictor of COVID-19 in younger children, whereas loss of taste or smell was the most determinant symptom in older children. Although the accuracy of the models was lower than expected, they can be used to provide a diagnosis when epidemiological data on the risk of exposure to COVID-19 is unknown.


Application of Glutathione as Anti-Oxidative and Anti-Aging Drugs.
Glutathione (GSH), an abundant tripeptidyl molecule, plays pivotal roles in protecting cells against oxidative stress-induced cellular damage and in detoxifying xenobiotics and drug metabolism. GSH is now entering a new era of therapeutic applications. Decreased GSH levels are associated with the common features of aging as well as of a wide range of pathological conditions, including neurodegenerative disorders. Notably, GSH depletion and/or alterations in its metabolism appear to be crucial in the onset of Parkinson's disease. Despite the fact that GSH is required for cell survival, the molecular mechanism that links GSH depletion to cell death remains poorly understood. Recently, considerable attention has been focused on a newly defined type of cell death: irondependent cell death, also referred to as "ferroptosis". The iron chelator deferoxamine nearly abolishes ferroptosis induced by inhibiting GSH synthesis or cystine uptake by the xCT transporter. Deferoxamine preferentially abrogates the intralysosomal accumulation of iron and inhibits oxidative stress-induced lysosomal membrane permeabilization and cell death. The use of GSH and a prodrug derived from it can be useful, since the dysfunction of the GSH redox system appears to cause a variety of diseases including neurodegenerative disorders. However, the effectiveness of GSH as a therapeutic agent is limited because of its low bioavailability. We also review trials that have been designed to cope with this difficulty; e.g. the use of precursors such as N-acetyl cysteine and chemical modification such as methylation.


Fetal Metabolic Stress Disrupts Immune Homeostasis and Induces Proinflammatory Responses in Human Immunodeficiency Virus Type 1- and Combination Antiretroviral Therapy-Exposed Infants.
Increased morbidity and fetal growth restriction are reported in uninfected children born to human immunodeficiency virus type 1 (HIV-1)-infected women treated with antiretroviral (ARV) therapy. Viruses and/or pharmacological interventions such as ARVs can induce metabolic stress, skewing the cell's immune response and restricting (cell) growth. Novel metabolomic techniques provided the opportunity to investigate the impact of fetal HIV-1 and combination ARV therapy (cART) exposure on the infants' immune metabolome. Peroxidized lipids, generated by reactive oxygen species, were increased in cART/HIV-1-exposed infants, indicating altered mitochondrial functioning. The lipid metabolism was further dysregulated with increased triglyceride species and a subsequent decrease in phospholipids in cART/HIV-1-exposed infants compared to control infants. Proinflammatory immune mediators, lysophospholipids as well as cytokines such as CXCL10 and CCL3, were increased whereas anti-inflammatory metabolites from the cytochrome P450 pathway were reduced in cART/HIV-1-exposed infants. Taken together, these data demonstrate that the fetal metabolism is impacted by maternal factors (cART and HIV-1) and skews physiological immune responses toward inflammation in the newborn infant.


Calcium phosphate content does not affect stone-free rate after percutaneous nephrolithotomy.
Recent research suggests that the stone-free rate for percutaneous nephrostolithotomy is lower in patients with calcium phosphate stones than in those with stones of other compositions. We reviewed our percutaneous nephrostolithotomy outcomes to investigate this unexplained finding. A total of 188 patients with sufficient data available for review underwent unilateral percutaneous nephrostolithotomy at our institution between September 2005 and May 2007. Patients were analyzed based on stone burden (including 2 cm or less, greater than 2 cm, partial staghorn calculus and complete staghorn calculus). Stones were also stratified by calcium phosphate content (0%, 1% to 10%, 11% to 60% and greater than 60%). To remain consistent with the previous study procedural failure was classified as greater than 2 mm residual stone fragments identified by unenhanced computerized tomography on postoperative day 1 regardless of the ultimate stone-free rate after secondary procedures. Multivariate logistic regression analysis was done to identify factors predicting a failed procedure. Of the patients 101 (54%) were male and 132 (71%) were recurrent stone formers. Overall 107 cases (57%) had calcium phosphate as a stone component and 37.8% were classified as failures. Increasing stone size was associated with a decreased stone-free rate (p = 0.009). The failure rate was 37%, 46.4%, 38.1% and 32.4% for patients with a 0%, 1% to 10%, 11% to 60% and greater than 60% calcium phosphate stone content (p = 0.68). On multivariate logistic regression analysis no association was noted between calcium phosphate content and greater than 2 mm residual stones (p = 0.67). Calcium phosphate stone composition does not predict a poor stone-free rate after percutaneous nephrostolithotomy.


Effects of somatostatin on acute pancreatitis induced in rats by injection of taurocholate and trypsin into a temporarily closed duodenal loop.
The effect of somatostatin on the course and severity of experimental pancreatitis was tested. Acute pancreatitis was induced in 210 Sprague-Dawley rats by injecting a 4.3% sodium taurocholate solution, saturated with trypsin, into a temporarily closed duodenal loop. Immediately after the end of the surgical procedure somatostatin or, alternatively, normal saline were administered as a bolus followed by continuous subcutaneous infusion for 9 h. Ninety rats (30 untreated, 30 saline-treated and 30 somatostatin-treated) were sacrificed 10 h after the induction of pancreatitis to assess the histologic severity of pancreatic lesions, the amount of peritoneal exudate and the circulating levels of amylase. In another 120 rats (40 untreated, 40 saline-treated and 40 drug-treated) the mortality rate was evaluated so that the histologic examination of the pancreas followed spontaneous death. In sacrificed animals somatostatin treatment lowered serum amylase levels and definitely improved pancreatic histopathology (edema, leucocyte infiltration and necrosis). The drug prevented the occurrence of severe necrosis in all treated animals. Somatostatin did not affect the mortality rate of pancreatitic rats (70%) although post-mortem histologic examination revealed significantly less pancreatic histopathology in drug-treated rats than in their controls.


Neuronal apoptosis in fatal familial insomnia.
The possibility that neuronal loss in prion diseases occurs through an apoptotic process has been postulated and is consistent with the lack of inflammation in these disorders. In order to test this hypothesis in FFI, in which neuronal loss is the predominant neuropathological feature, we examined samples of thalamus, basal ganglia, cerebral cortex, cerebellum and medulla from 10 subjects with FFI. All the patients had the characteristic 178 N mutation of the PrP gene. Eight subjects were homozygous methionine/methionine at codon 129 and 2 were heterozygous methionine/valine. Apoptotic neurons were identified by in situ end labelling in all the FFI cases and in none of the controls. They were mostly found in damaged regions and their presence and abundance seemed to correlate closely with the neuronal loss. They were particularly abundant in the thalamus and medullary olives. In heterozygous cases who had a longer disease duration and more widespread cerebral changes, apoptotic neurons were also found in the neocortex and striatum. The abundance of apoptotic neurons also correlated well with microglial activation as demonstrated by the expression of major histocompatibility complex class II antigens. PrPres immunostaining was almost invariably negative, consistent with previous data showing the lack of obvious correlation between neuronal loss and PrPres deposits in prion diseases.


Geometric diagnostics of complex patterns: spiral defect chaos.
Motivated by the observation of spiral patterns in a wide range of physical, chemical, and biological systems, we present an automated approach that aims at characterizing quantitatively spiral-like elements in complex stripe-like patterns. The approach provides the location of the spiral tip and the size of the spiral arms in terms of their arc length and their winding number. In addition, it yields the number of pattern components (Betti number of order 1), as well as their size and certain aspects of their shape. We apply the method to spiral defect chaos in thermally driven Rayleigh-Benard convection and find that the arc length of spirals decreases monotonically with decreasing Prandtl number of the fluid and increasing heating. By contrast, the winding number of the spirals is nonmonotonic in the heating. The distribution function for the number of spirals is significantly narrower than a Poisson distribution. The distribution function for the winding number shows approximately an exponential decay. It depends only weakly on the heating, but strongly on the Prandtl number. Large spirals arise only for larger Prandtl numbers (Pr approximately > 1). In this regime the joint distribution for the spiral length and the winding number exhibits a three-peak structure, indicating the dominance of Archimedean spirals of opposite sign and relatively straight sections. For small Prandtl numbers the distribution function reveals a large number of small compact pattern components.


Scientism and economism in the regulation of health care.
As health care costs continue their apparently relentless rise, it seems to be universally perceived that the United States and western Europe are gripped by a cost crisis. To resolve the apparent crisis, U.S. and western European governments and third-party payers are turning increasingly to a new positivist discipline, called health services research, for which neoclassical health economics is the dominant discourse. However this discipline may actually reinforce the strength of biomedical positivism and the concomitant technological imperative. Like biomedicine, health services research is technologically driven, dependent on "advances" that generate more comprehensive and therefore more "accurate" data. Accordingly, just as biomedicine causes health care workers and patients to depend on technologies for diagnosis and treatment, health services research instills in the body politic dependence on technocratically conceived solutions for political problems. Moreover, because biomedicine and health services research share positivist epistemic and methodological premises, both objectify the subjects they study, abstract those subjects from context, and thereby ignore the cultural dimensions of the problems at hand. Rather than inculcate an ethic and practice in which medicine focuses on the meaning of illness for a life, a cultural phenomenon, this form of positivism strengthens the tendency to reject meaning in favor of the causes and course of disease and the abstracted probability of its occurrence. Accordingly health services research and the forms of regulation with which it is allied threaten to overwhelm the medical humanities movement. Furthermore this scientism precludes the institutionalization of political forums in which we can deliberate on the meaning of medicine, health, and death in our lives.


EGFR High Expression, but not KRAS Status, Predicts Sensitivity of Pancreatic Cancer Cells to Nimotuzumab Treatment In Vivo.
Nimotuzumab is shown to be efficacious in advanced pancreatic cancer treatment, but its predictive marker has not been established. To investigate the impact of EGFR and KRAS status on antitumor efficacy of nimotuzumab and to explore its underlying mechanism. EGFR expressions of pancreatic cancer cell lines, BxPC3, Panc-1, and Patu-8988, were analyzed by Western blot and immunocytochemistry, and KRAS status was determined by gene sequencing. Anti-tumor effect of nimotuzumab were evaluated in vitro and in vivo. The expressions of related molecules in EGFR pathway and IL-6 was analyzed by Western blot, immunohistochemistry, and/or real-time PCR. BxPC3 cells had wild type KRAS and high-level EGFR; Panc-1 cells had mutant KRAS (G13A) and low-level EGFR; Patu-8988 cells had mutant KRAS (G12V) and high-level EGFR. Nimotuzumab did not affect cell proliferation or apoptosis in vitro. Growth of BxPC3 and Patu-8988 xenografts were significantly inhibited by nimotuzumab, but not Panc-1 xenografts, compared with that of the control group. Expression of EGFR in BxPC3 and Patu-8988 xenografts was significantly reduced by nimotuzumab. The IL-6 expression in BxPC3 and Patu-8988 xenografts was higher than that in Panc-1 xenografts in the control group, and was significantly reduced by nimotuzumab. Pancreatic cancer cells with EGFR high expression were more sensitive to nimotuzumab in vivo. KRAS status had no impact on anti-tumor efficacy of nimotuzumab in pancreatic cancer cells.


4-O-Acetyl-sialic acid (Neu4,5Ac2) in acidic milk oligosaccharides of the platypus (Ornithorhynchus anatinus) and its evolutionary significance.
Monotremes (echidnas and platypus) retain an ancestral form of reproduction: egg-laying followed by secretion of milk onto skin and hair in a mammary patch, in the absence of nipples. Offspring are highly immature at hatching and depend on oligosaccharide-rich milk for many months. The primary saccharide in long-beaked echidna milk is an acidic trisaccharide Neu4,5Ac2(α2-3)Gal(β1-4)Glc (4-O-acetyl 3'-sialyllactose), but acidic oligosaccharides have not been characterized in platypus milk. In this study, acidic oligosaccharides purified from the carbohydrate fraction of platypus milk were characterized by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and (1)H-nuclear magnetic resonance spectroscopy. All identified structures, except Neu5Ac(α2-3)Gal(β1-4)Glc (3'-sialyllactose) contained Neu4,5Ac2 (4-O-acetyl-sialic acid). These include the trisaccharide 4-O-acetyl 3'-sialyllactose, the pentasaccharide Neu4,5Ac2(α2-3)Gal(β1-4)GlcNAc(β1-3)Gal(β1-4)Glc (4-O-acetyl-3'-sialyllacto-N-tetraose d) and the hexasaccharide Neu4,5Ac2(α2-3)Gal(β1-4)[Fuc(α1-3)]GlcNAc(β1-3)Gal(β1-4)Glc (4-O-acetyl-3'-sialyllacto-N-fucopentaose III). At least seven different octa- to deca-oligosaccharides each contained a lacto-N-neohexaose core (LNnH) and one or two Neu4,5Ac2 and one to three fucose residues. We conclude that platypus milk contains a diverse (≥ 20) array of neutral and acidic oligosaccharides based primarily on lactose, lacto-N-neotetraose (LNnT) and LNnH structural cores and shares with echidna milk the unique feature that all identified acidic oligosaccharides (other than 3'-sialyllactose) contain the 4-O-acetyl-sialic acid moiety. We propose that 4-O-acetylation of sialic acid moieties protects acidic milk oligosaccharides secreted onto integumental surfaces from bacterial hydrolysis via steric interference with bacterial sialidases. This may be of evolutionary significance since taxa ancestral to monotremes and other mammals are thought to have secreted milk, or a milk-like fluid containing oligosaccharides, onto skin surfaces.


An Experimental Model of Neuromyelitis Optica Spectrum Disorder-Optic Neuritis: Insights Into Disease Mechanisms.
Background: Optic neuritis (ON) is a common inflammatory optic neuropathy, which often occurs in neuromyelitis optica spectrum disease (NMOSD). An experimental model of NMOSD-ON may provide insight into disease mechanisms. Objective: To examine the pathogenicity of autoantibodies targeting the astrocyte water channel aquaporin-4 [aquaporin-4 (AQP4)-immunoglobulin G (AQP4-IgG)] in the optic nerve. Materials and Methods: Purified IgG from an AQP4-IgG-positive NMOSD-ON patient was together with human complement (C) given to wild-type (WT) and type I interferon (IFN) receptor-deficient mice (IFNAR1-KO) as two consecutive intrathecal injections into cerebrospinal fluid via cisterna magna. The optic nerves were isolated, embedded in paraffin, cut for histological examination, and scored semi-quantitatively in a blinded fashion. In addition, optic nerves were processed to determine selected gene expression by quantitative real-time PCR. Results: Intrathecal injection of AQP4-IgG+C induced astrocyte pathology in the optic nerve with loss of staining for AQP4 and glial fibrillary acidic protein (GFAP), deposition of C, and demyelination, as well as upregulation of gene expression for interferon regulatory factor-7 (IRF7) and CXCL10. Such pathology was not seen in IFNAR1-KO mice nor in control mice. Conclusion: We describe induction of ON in an animal model for NMOSD and show a requirement for type I IFN signaling in the disease process.


Insurance disparities in the outcomes of spinal cord stimulation surgery.
The Affordable Care Act aims to expand health insurance and to help narrow existing health care disparities. Medicaid patients have previously been noted to be at an increased risk for impaired access to health care, delayed medical treatment, and the receipt of substandard care. Conversely, those with commercial insurance may be subject to overtreatment. The goal of this study was to evaluate how Medicaid versus commercial insurance status affects outcomes following spinal cord stimulation (SCS) surgery. A retrospective cohort study of 13,774 patients underwent either percutaneous or paddle permanent SCS implantation, selected from the Thomson Reuter's MarketScan database between 2000 and 2009. Patients were characterized by age at initial procedure, gender, baseline comorbidity burden, procedure-associated diagnosis code, follow-up, and type of insurance (Medicaid vs. commercial insurance). Outcome measures included probability of reoperation, timing and type of reoperation, presence of postoperative complications (immediate, 30 days, and 90 days), and overall utilization of health resources postoperatively. Multivariate analysis was performed comparing the relative effect of insurance status on outcomes following initial surgery. Medicaid patients had greater healthcare resource utilization as measured by medications prescribed, emergency department visits, and length of stay; however, commercially insured patients had significantly higher overall costs ($110,908 vs. $64,644, p < 0.0001). Commercial and Medicaid patients did not significantly differ in their complication rates during the index hospitalization or at 30 days or 90 days postoperatively. The group were also not significantly different in their two-year reoperation rates (7.32% vs. 5.06%, p = 0.0513). There are substantial insurance disparities that affect healthcare utilization and overall cost following SCS. Efforts for national healthcare reform should examine system factors that will reduce socioeconomic disparities in outcomes following SCS.


Nickel-titanium coil springs and repelling magnets: a comparison of two different intra-oral molar distalization techniques.
The aim of this study was to compare two intra-oral molar distalization procedures, involving 15 cases with Class II molar relationships. Magnetic devices were applied to the upper right first molars in each case, while nickel-titanium coil springs were used against the upper left first molars, for a period of 3 months. Measurements were made from lateral cephalograms and from photocopies of models taken before and after the distalization procedure. Although upper molar distalization was achieved with ease in both techniques, nickel-titanium coil springs were found to be the more effective in terms of movement achieved.


Spectrum of variants in a large Chinese Gitelman syndrome cohort.
Gitelman syndrome (GS) is caused by SLC12A3 biallelic variants. A previous study showed that large rearrangements (LRGs) of SLC12A3 accounted for the low sensitivity of genetic testing. However, a systematic screening for LRGs in Chinese GS patients is lacking. Massively parallel sequencing (MPS) and multiplex ligation-dependent probe amplification (MLPA) were performed to sequence the genomic DNA of patients with clinically diagnosed GS. Of 165 index cases, MPS identified 151 cases with two or more affected alleles and 14 cases with one variant allele. LRGs were detected by MLPA in 20 out of 27 cases, including 15 cases with suspected LRGs by MPS. Among these 20 cases with LRGs, the results of MPS and MLPA were identical in only 8 cases. Additional LRGs in 6 cases were detected by MLPA alone. In 6 cases, E4_E6del was identified by MPS, while E4_E5del and Intron6del were identified by MLPA. Among the 102 distinct variants, 30 are novel. LRGs were found in 20 cases (12.1%). LRGs were found in 12.1% of our Chinese GS patients cohort. We show that MPS and MLPA are two complementary techniques with the ability to improve the diagnostic yield of GS.


Physiological demands of standing and wheelchair fencing in able-bodied fencers.
The purpose of this study was to determine the cardiorespiratory demands of standing and wheelchair (seated) fencing in a group of able-bodied fencers during simulated competitive bouts. Participants were ten male able-bodied fencers of regional level with previous training experience in wheelchair fencing. After a standardized warm-up, participants performed two series of simulated competitive épée bouts (5 and 15 touches) in a random order, either while standing or while sitting in a wheelchair. Expired gas was analyzed for oxygen consumption (V̇O2) and respiratory exchange ratio (RER) and heart rate were continually monitored. Energy expenditure (EE) was subsequently calculated. V̇O2, HR and EE peak responses were greater during standing than seated fencing (P<0.05). Mean V̇O2 during all ST bouts (5- and 15-touch) was 43% greater than in wheelchair fencing (44.2±7.8 vs. 25.1±5.4 mL/kg/min). Mean HR during the standing 5- and 15-touch bouts was 91±20% and 84±7%, respectively, of that recorded during the seated bouts. HR, V̇O2 and EE data also indicated that the 15-touch bouts were more physiologically demanding than the 5-touch bouts (P<0.01). The HR-V̇O2 relationship was similar between both fencing modes. The duration of the 5- and 15-touch bouts were shorter for the seated than the standing bouts (P<0.01). The physiological demands of wheelchair fencing are lower than those for standing fencing. Furthermore, the physiology of 5 versus 15-touch bouts, similar to those undertaken in fencing competition, also differs.


Ecallantide: a plasma kallikrein inhibitor for the treatment of acute attacks of hereditary angioedema.
Hereditary angioedema (HAE) is a debilitating, potentially fatal disease characterized by variable and unpredictable acute attacks of swelling affecting the subcutaneous tissue and mucosa. It is an autosomal dominant disorder resulting from a genetic deficiency of functional C1-esterase inhibitor. Available treatments include long-term prophylaxis, short-term prophylaxis and treatment of acute attacks. Ecallantide is a novel, specific and potent inhibitor of plasma kallikrein that was recently approved in the United States for the treatment of acute attacks of HAE in patients aged 16 years and older. In two phase III clinical trials, the subcutaneous administration of 30 mg ecallantide resulted in significantly greater symptom improvement than placebo for acute attacks of HAE. Ecallantide was generally well tolerated throughout the clinical development program. The main safety concern following ecallantide treatment is hypersensitivity reactions, including anaphylaxis. A Risk Evaluation and Management Strategy (REMS) has been implemented to minimize this risk and a long-term observational safety study is currently under way to collect more information about hypersensitivity and immunogenicity. Ecallantide represents a novel treatment option for patients with HAE.


Synthesis, biological evaluation, and structure-activity relationships of N-benzoyl-2-hydroxybenzamides as agents active against P. falciparum (K1 strain), Trypanosomes, and Leishmania.
In our efforts to identify novel chemical scaffolds for the development of new antiprotozoal drugs, a compound library was screened against Toxoplasma gondii tachyzoites with activity discovered for N-(4-ethylbenzoyl)-2-hydroxybenzamide 1a against T. gondii as described elsewhere. Synthesis of a compound set was guided by T. gondii SAR with 1r found to be superior for T. gondii , also active against Thai and Sierra Leone strains of Plasmodium falciparum , and with superior ADMET properties as described elsewhere. Herein, synthesis methods and details of the chemical analysis of the compounds in this series are described. Further, this series of N-benzoyl-2-hydroxybenzamides was repurposed for testing against four other protozoan parasites: Trypanosoma brucei rhodesiense , Trypanosoma cruzi , Leishmania donovani , and P. falciparum (K1 isolate). Structure-activity analyses led to the identification of compounds in this set with excellent antileishmanial activity (compound 1d). Overall, compound 1r was the best and had activity 21-fold superior to that of the standard antimalarial drug chloroquine against the K1 P. falciparum isolate.


Force-dependent chemical kinetics of disulfide bond reduction observed with single-molecule techniques.
The mechanism by which mechanical force regulates the kinetics of a chemical reaction is unknown. Here, we use single-molecule force-clamp spectroscopy and protein engineering to study the effect of force on the kinetics of thiol/disulfide exchange. Reduction of disulfide bonds through the thiol/disulfide exchange chemical reaction is crucial in regulating protein function and is known to occur in mechanically stressed proteins. We apply a constant stretching force to single engineered disulfide bonds and measure their rate of reduction by DTT. Although the reduction rate is linearly dependent on the concentration of DTT, it is exponentially dependent on the applied force, increasing 10-fold over a 300-pN range. This result predicts that the disulfide bond lengthens by 0.34 A at the transition state of the thiol/disulfide exchange reaction. Our work at the single bond level directly demonstrates that thiol/disulfide exchange in proteins is a force-dependent chemical reaction. Our findings suggest that mechanical force plays a role in disulfide reduction in vivo, a property that has never been explored by traditional biochemistry. Furthermore, our work also indicates that the kinetics of any chemical reaction that results in bond lengthening will be force-dependent.


Is there a relationship between blood pressure values and dementia?
The relationship between arterial hypertension and cognitive decline, two among the conditions with higher prevalence in the elderly population, has gained significant interest, in the scientific community, during the last few years, stemming from the numerous epidemiologic, experimental, and therapeutic evidences suggesting a non-casual correlation between the two conditions. In fact, the brain, for its substantial metabolic and functional complexity, is more susceptible to the harmful effect of high blood pressure than the other target organs. Chronic ischaemic impairment, microvascular damage, and neurodegenerative phenomena are the likely pathophysiologic basis for the correlation between hypertension and cognitive decline. Vascular dementia and Alzheimer's disease, the two prominent forms of senile dementia, seem to represent the end result of the chronic exposure, during the lifetime, to harmful stimuli, among which the most relevant are the cardiovascular risk factors, at least from an epidemiological perspective. Evidences from interventional studies, although limited, seems to support the concept that to limit the spread of senile dementia, the early optimization of the control of cardiovascular risk factors, first and foremost hypertension, is crucial. The occurrence of a variable degree of mental decline, till overt dementia, in the hypertensive patient, represents the final step of a pathophysiologic process that began many years before. There is, then, the clear opportunity to control the pathophysiologic mechanisms leading to cognitive decline in the hypertensive patient.


TiO2 coating promotes human mesenchymal stem cell proliferation without the loss of their capacity for chondrogenic differentiation.
Human mesenchymal stem cells (hMSCs) are used in applications, which may require a large amount of cells; therefore, efficient expansion of the cells is desired. We studied whether TiO2 coating on plastic cell culture dishes could promote proliferation of hMSCs without adverse effects in chondrogenic differentiation. TiO2-films were deposited on polystyrene dishes and glass coverslips using an ultrashort pulsed laser deposition technique. Human MSCs from three donors were expanded on them until 95% confluence, and the cells were evaluated by morphology, immunocytochemistry and quantitative RT-PCR (qRT-PCR). The chondrogenic differentiation in pellets was performed after cultivation on TiO2-coated dishes. Chondrogenesis was evaluated by histological staining of proteoglycans and type II collagen, and qRT-PCR. Human MSC-associated markers STRO-1, CD44, CD90 and CD146 did not change after expansion on TiO2-coated coverslips. However, the cell number after a 48h-culture period was significantly higher on TiO2-coated culture dishes. Importantly, TiO2 coating caused no significant differences in the proteoglycan and type II collagen staining of the pellets, or the expression of chondrocyte-specific genes in the chondrogenesis assay. Thus, the proliferation of hMSCs could be significantly increased when cultured on TiO2-coated dishes without weakening their chondrogenic differentiation capacity. The transparency of TiO2-films allows easy monitoring of the cell growth and morphology under a phase-contrast microscope.


Human leukocyte functions and the U.S. safety standard for exposure to radio-frequency radiation.
Human mononuclear leukocytes were exposed to microwaves at energies relevant to current public safety recommendations. No detectable effects on viability or function of the leukocytes resulted from exposure to microwaves at specific absorption rates up to 4 milliwatts per milliliter. The results were highly reproducible and provided no evidence that current safety standard recommendations are inappropriate insofar as leukocyte function is concerned.