Antagonism of 4-substituted 1,4-dihydropyridine-3,5-dicarboxylates toward voltage-dependent L-type Ca2+ channels Ca V 1.3 and Ca V 1.2.
L-type Ca(2+) channels in mammalian brain neurons have either a Ca(V)1.2 or Ca(V)1.3 pore-forming subunit. Recently, it was shown that Ca(V)1.3 Ca(2+) channels underlie autonomous pacemaking in adult dopaminergic neurons in the substantia nigra pars compacta, and this reliance renders them sensitive to toxins used to create animal models of Parkinson's disease. Antagonism of these channels with the dihydropyridine antihypertensive drug isradipine diminishes the reliance on Ca(2+) and the sensitivity of these neurons to toxins, pointing to a potential neuroprotective strategy. However, for neuroprotection without an antihypertensive side effect, selective Ca(V)1.3 channel antagonists are required. In an attempt to identify potent and selective antagonists of Ca(V)1.3 channels, 124 dihydropyridines (4-substituted-1,4-dihydropyridine-3,5-dicarboxylic diesters) were synthesized. The antagonism of heterologously expressed Ca(V)1.2 and Ca(V)1.3 channels was then tested using electrophysiological approaches and the FLIPR Calcium 4 assay. Despite the large diversity in substitution on the dihydropyridine scaffold, the most Ca(V)1.3 selectivity was only about twofold. These results support a highly similar dihydropyridine binding site at both Ca(V)1.2 and Ca(V)1.3 channels and suggests that other classes of compounds need to be identified for Ca(V)1.3 selectivity.


Episodic generation can cause semantic forgetting: retrieval-induced forgetting of false memories.
In two experiments, we tested whether false recognition and false recall were prone to retrieval-induced forgetting, using the retrieval practice paradigm (Anderson, Bjork, & Bjork, 1994). Participants encoded lists of cue-target word pairs associated with a nonpresented, critical theme word and then engaged in retrieval practice for half of the word pairs from half of the lists. As expected, unpracticed targets from practiced lists were recognized (Experiment 1) and recalled (Experiment 2) less well than those from unpracticed lists. In addition, false recognition and false recall of critical items associated with practiced lists was lower than false recognition and false recall of items associated with unpracticed lists. We argue that false memories are prone to inhibitory mechanisms engendered by the retrieval practice paradigm. The results are consistent with the claim that semantically activated critical themes interfere with the episodic retrieval of list words and that inhibition decreases the activation level of these interfering memory representations during retrieval practice.


Does Charge Balancing Ensure the Safety of the Electrical Stimulation and Is It Power Efficientƒ.
Safety issues are the most important concern in electrical stimulation. Equating the charge in the anodic and cathodic phases, namely charge balancing or charge equalizing, is a well-known method to avoid tissue damage and/or electrode corrosion. The electrode-tissue interface is not ideal in practice and with a charge-balanced waveform, the electrode voltage becomes more positive compared to the pre-pulse value and corrosion happens. In this paper, we show that a charge balancer ensures the safety of stimulation if the rate of the irreversible Faradaic reactions is negligible, or when the pulse width of the stimulation phases is not comparable to the time constant of the electrode-tissue interface. Furthermore, charge balancing is studied with mathematical modeling for different types of tissue models, and the results are used to show the conditions that charge balancing does not ensure the safety of the electrical stimulation, and employing charge balancing not only increases the power consumption of the electrical-stimulation systems but also increases the rate of the electrode corrosion in these conditions. The main goal of this paper is to show that a charge controller is a general solution for ensuring the safety of the electrical stimulation, with an efficient, not excessive, amount of charge for the reversal phase and should be employed instead of charge balancers in generic stimulators.


Neural mechanisms underlying concurrent listening of simultaneous speech.
Can we identify what two people are saying at the same time? Although it is difficult to perfectly repeat two or more simultaneous messages, listeners can report information from both speakers. In a concurrent/divided listening task, enhanced attention and segregation of speech can be required rather than selection and suppression. However, the neural mechanisms of concurrent listening to multi-speaker concurrent speech has yet to be clarified. The present study utilized functional magnetic resonance imaging to examine the neural responses of healthy young adults listening to concurrent male and female speakers in an attempt to reveal the mechanism of concurrent listening. After practice and multiple trials testing concurrent listening, 31 participants achieved performance comparable with that of selective listening. Furthermore, compared to selective listening, concurrent listening induced greater activation in the anterior cingulate cortex, bilateral anterior insula, frontoparietal regions, and the periaqueductal gray region. In addition to the salience network for multi-speaker listening, attentional modulation and enhanced segregation of these signals could be used to achieve successful concurrent listening. These results indicate the presence of a potential mechanism by which one can listen to two voices with enhanced attention to saliency signals.


Functional dissection of an innate immune response by a genome-wide RNAi screen.
The innate immune system is ancient and highly conserved. It is the first line of defense and the only recognizable immune system in the vast majority of metazoans. Signaling events that convert pathogen detection into a defense response are central to innate immunity. Drosophila has emerged as an invaluable model organism for studying this regulation. Activation of the NF-kappaB family member Relish by the caspase-8 homolog Dredd is a central, but still poorly understood, signaling module in the response to gram-negative bacteria. To identify the genes contributing to this regulation, we produced double-stranded RNAs corresponding to the conserved genes in the Drosophila genome and used this resource in genome-wide RNA interference screens. We identified numerous inhibitors and activators of immune reporters in a cell culture model. Epistatic interactions and phenotypes defined a hierarchy of gene action and demonstrated that the conserved gene sickie is required for activation of Relish. We also showed that a second gene, defense repressor 1, encodes a product with characteristics of an inhibitor of apoptosis protein that inhibits the Dredd caspase to maintain quiescence of the signaling pathway. Molecular analysis revealed that Defense repressor 1 is upregulated by Dredd in a feedback loop. We propose that interruption of this feedback loop contributes to signal transduction.


Bioengineered Vascular Model of Foam Cell Formation.
Foam cell formation is a complex blood vessel pathology, which is characterized by a series of events, including endothelium dysfunction, inflammation, and accumulation of immune cells underneath the blood vessel walls. Novel bioengineered models capable of recapitulating these events are required to better understand the complex pathological processes underlying the development of foam cell formation and, consequently, advanced bioengineered platforms for screening drugs. Here, we generated a microfluidic blood vessel model, incorporating a three-dimensional (3D) extracellular matrix coated with an endothelial layer. This system enables us to perform experiments under a dynamic microenvironment that recapitulates the complexities of the native vascular regions. Using this model, we studied the effectors that regulate monocyte adhesion and migration, as well as foam cell formation inside vessel walls. We found that monocyte adhesion and migration are regulated by both the endothelium and monocytes themselves. Monocytes migrated into the extracellular matrix only when endothelial cells were cultured in the vessel model. In addition, the exposure of an endothelial layer to tumor necrosis factor α (TNF-α) and low shear stress both increased monocyte migration into the subendothelial space toward the matrix. Furthermore, we demonstrated the process of foam cell formation, 3 days after transmigration of peripheral blood mononuclear cells (PBMCs) into the vessel wall. We showed that pre-exposure of PBMCs to high shear rates increases their adhesion and migration through the TNF-α-treated endothelium but does not affect their capacity to form foam cells. The versatility of our model allows for mechanistic studies on foam cell formation under customized pathological conditions.


Effects of Moisture and Particle Size on Quantitative Determination of Total Organic Carbon (TOC) in Soils Using Near-Infrared Spectroscopy.
Near-Infrared Spectroscopy is a cost-effective and environmentally friendly technique that could represent an alternative to conventional soil analysis methods, including total organic carbon (TOC). Soil fertility and quality are usually measured by traditional methods that involve the use of hazardous and strong chemicals. The effects of physical soil characteristics, such as moisture content and particle size, on spectral signals could be of great interest in order to understand and optimize prediction capability and set up a robust and reliable calibration model, with the future perspective of being applied in the field. Spectra of 46 soil samples were collected. Soil samples were divided into three data sets: unprocessed, only dried and dried, ground and sieved, in order to evaluate the effects of moisture and particle size on spectral signals. Both separate and combined normalization methods including standard normal variate (SNV), multiplicative scatter correction (MSC) and normalization by closure (NCL), as well as smoothing using first and second derivatives (DV1 and DV2), were applied to a total of seven cases. Pretreatments for model optimization were designed and compared for each data set. The best combination of pretreatments was achieved by applying SNV and DV2 on partial least squares (PLS) modelling. There were no significant differences between the predictions using the three different data sets (p < 0.05). Finally, a unique database including all three data sets was built to include all the sources of sample variability that were tested and used for final prediction. External validation of TOC was carried out on 16 unknown soil samples to evaluate the predictive ability of the final combined calibration model. Hence, we demonstrate that sample preprocessing has minor influence on the quality of near infrared spectroscopy (NIR) predictions, laying the ground for a direct and fast in situ application of the method. Data can be acquired outside the laboratory since the method is simple and does not need more than a simple band ratio of the spectra.


Polypharmacy Is Associated With Amiodarone-Induced Hypothyroidism.
Background: Amiodarone is rich in iodine, so in clinical practice amiodarone-induced hypothyroidism (AIH) is a major side effect. This drug is used in patients with arrhythmias, especially atrial fibrillation, the most common sustained arrhythmia. Polypharmacy, which can result in complex drug-drug interactions, occurs in more than 70% of the patients with atrial fibrillation. Therefore, polypharmacy may be involved in the expression of AIH. In this study, we investigated the association between polypharmacy and AIH. Methods: We conducted a retrospective study using data from January 2006 to May 2020 collected from a large, organized database of prescriptions constructed by the Japan Medical Information Research Institute, Inc. (Tokyo, Japan). To investigate the association between number of prescribed drugs with amiodarone and AIH, we divided patients into two groups: polypharmacy (≥ 5 prescribed drugs) and non-polypharmacy (< 5 prescribed drugs). We then performed a sequence symmetry analysis on the two groups: incident thyroxine after incident amiodarone and incident thyroxine before incident amiodarone. Finally, we conducted a case-control study on two further groups: those prescribed thyroxine after incident amiodarone (AIH group; n=555) and those not prescribed thyroxine after incident amiodarone (non-AIH group; n=6,192). Results: Sequence symmetry analysis revealed a significant association between amiodarone and thyroxine in both the polypharmacy and non-polypharmacy groups. The ranges for the adjusted sequence ratio in the two groups were 12.0-16.7 and 7.3-9.0, respectively. The case-control study showed that ≥5 prescribed drugs at the first prescription of amiodarone were found to significantly increase the odds of AIH (odds ratio: 1.48, 95% confidence interval: 1.18-1.84). Conclusion: Polypharmacy was suggested as an independent risk factor for AIH. Careful assessment of the appropriateness of prescription is warranted.


[Confocal microscopy of the cornea. Communication 1. The normal morphological pattern].
The paper presents the results of life-time confocal microscopy using a Confoscan-4 confocal microscope (Nidek). The study was conducted on 10 healthy volunteers (20 eyes), which evaluated the specific features of the normal morphological pattern of the cornea. The morphological features of three types of anterior corneal epithelial (basal, wing, and spiny) cells were revealed. The possibilities of visualizing subbasal and stromal nerve fibers were assessed. Corneal stromal layers (these being anterior, anteromedian, median, and posterior) differing in the density of cellular elements, their shape, and orientation) were conventionally identified. In health, Bowman's and Descemet's membranes were not visualized as they were transparent and failed to reflect light. The posterior corneal epithelium was microscopically presented by a layer of light hexagonal or polygonal squamous cells with well-defined outlines. Thus, life-time corneal confocal microscopy is a highly informative noninvasive study.


Can we use gonadotropin plasma concentration as surrogate marker for BMI-related incomplete estrogen suppression in breast cancer patients receiving anastrozole?
BMI has been suggested to impact on estrogenic activity in patients receiving anastrozole resulting in a reduced treatment efficacy in obese women. Current evidence in this regard is controversially discussed. Since estradiol is inversely correlated with gonadotropins it can be assumed that an impact of BMI is also reflected by gonadotropin plasma concentrations. We aim at investigating the impact of BMI on the hormonal state of breast cancer (BC) patients receiving anastrozole indicated by LH, FSH and SHBG as well as estradiol. We determined gonadotropin-, estradiol- and anastrozole- serum concentrations from postmenopausal, early stage breast cancer patients receiving upfront anastrozole within routine after care. Gonadotropin plasma concentrations were derived from the routine laboratory examination report. A liquid chromatography tandem mass spectrometry method was used for the measurement of anastrozole serum concentrations. BMI was assessed within the routine after-care check-up. The overall sample comprised 135 BC patients with a mean age of 65.3 years. BMI was significantly correlated with LH, FSH and SHBG. This association was neither influenced by age nor by anastrozole serum concentrations according to the regression model. Despite aromatase inhibition 12% of patients had detectable estrogen levels in routine quantification. Obese women have an altered hormonal situation compared to normally weight women under the same dose of anastrozole. Our study findings are a further indicator for the relevance of BMI in regard of anastrozole metabolism and possible estrogenic activity indicated by gonadotropin plasma level.


Emission estimation and prioritization of veterinary pharmaceuticals in manure slurries applied to soil.
Veterinary pharmaceuticals (VPs) are emitted into the environment and transfer to groundwater and surface water is diffuse and complex, whereas actual information on the fate is frequently limited. For 17 VPs of potential concern in the Netherlands, we assessed sources and emission due to animal slurry applications to soil. Hence, we examined the use of VPs in four livestock sectors in the Netherlands for 2015-2018, and quantified animal excretion rates and dissipation during slurry storage. For almost all VPs, administrated quantities to the animals during the period 2015-2018 decreased. VP concentrations during a storage period of six months could decrease between 10 and 98% depending on the compound. Predicted concentrations of VPs in slurries after storage compared well with measured concentrations in the literature. Based on the storage model outcomes, we developed a residue indicator, that quantifies the potential for residues in applied slurry. This indicator agrees well with the most frequently detected VPs in the Dutch slurries, and is therefore useful to prioritize measures aiming at reducing VP emissions into the environment.


Psychometric properties of the Oswestry Disability Index.
The aim of this study was to investigate the psychometric properties of the Oswestry Disability Index (ODI) in a large cross-sectional cohort of individuals with chronic low back pain by defining its internal consistency, construct structure and validity, and its ability to differentiate between different degrees of functional limitation. A total of 837 consecutive outpatient patients with low back pain were studied. The internal consistency of ODI was assessed by Cronbach's α, construct structure by exploratory factor analysis, construct validity by confirmatory factor analysis, and discrimination was determined by item response theory analysis. The ODI showed good internal consistency (α=0.85). Explanatory factor analysis showed that ODI is a unidimensional test measuring functional level and nothing else. The confirmatory factor analysis showed that the standardized regression weights of all ODI items were relatively high, varying from 0.5 to 0.7. The item response theory analysis suggested that eight out of 10 ODI items have a close to perfect ability to measure functional limitations in accordance with the actual severity of disability experienced by the respondents. Discrimination of all the items was high to perfect (1.08-2.01). The test characteristic and test information curves showed that the discriminative ability of the ODI is superior at higher levels of disability. The present data showed that the ODI is an internally consistent, unidimensional scale with overall excellent construct validity and ability to discriminate the severity of functional disability. The analysis suggests that the ODI may better distinguish between the relative degrees of function at above-average disability levels.


Imaging Strategies for Suspected Acute Cranial Shunt Failure: A Cost-Effectiveness Analysis.
We compared cost-effectiveness of cranial computed tomography (CT), fast sequence magnetic resonance imaging (fsMRI), and ultrasonography measurement of optic nerve sheath diameter (ONSD) for suspected acute shunt failure from the perspective of a health care organization. We modeled 4 diagnostic imaging strategies: (1) CT scan, (2) fsMRI, (3) screening ONSD by using point of care ultrasound (POCUS) first, combined with CT, and (4) screening ONSD by using POCUS first, combined with fsMRI. All patients received an initial plain radiographic shunt series (SS). Short- and long-term costs of radiation-induced cancer were assessed with a Markov model. Effectiveness was measured as quality-adjusted life-years. Utilities and inputs for clinical variables were obtained from published literature. Sensitivity analyses were performed to evaluate the effects of parameter uncertainty. At a previous probability of shunt failure of 30%, a screening POCUS in patients with a normal SS was the most cost-effective. For children with abnormal SS or ONSD measurement, fsMRI was the preferred option over CT. Performing fsMRI on all patients would cost $269 770 to gain 1 additional quality-adjusted life-year compared with POCUS. An imaging pathway that involves CT alone was dominated by ONSD and fsMRI because it was more expensive and less effective. In children with low pretest probability of cranial shunt failure, an ultrasonographic measurement of ONSD is the preferred initial screening test. fsMRI is the more cost-effective, definitive imaging test when compared with cranial CT.


Recent developments in quantitative proteomics.
Proteomics is the study of proteins on a large scale, encompassing the many interests scientists and physicians have in their expression and physical properties. Proteomics continues to be a rapidly expanding field, with a wealth of reports regularly appearing on technology enhancements and scientific studies using these new tools. This review focuses primarily on the quantitative aspect of protein expression and the associated computational machinery for making large-scale identifications of proteins and their post-translational modifications. The primary emphasis is on the combination of liquid chromatography-mass spectrometry (LC-MS) methods and associated tandem mass spectrometry (LC-MS/MS). Tandem mass spectrometry, or MS/MS, involves a second analysis within the instrument after a molecular dissociative event in order to obtain structural information including but not limited to sequence information. This review further focuses primarily on the study of in vitro digested proteins known as bottom-up or shotgun proteomics. A brief discussion of recent instrumental improvements precedes a discussion on affinity enrichment and depletion of proteins, followed by a review of the major approaches (label-free and isotope-labeling) to making protein expression measurements quantitative, especially in the context of profiling large numbers of proteins. Then a discussion follows on the various computational techniques used to identify peptides and proteins from LC-MS/MS data. This review article then includes a short discussion of LC-MS approaches to three-dimensional structure determination and concludes with a section on statistics and data mining for proteomics, including comments on properly powering clinical studies and avoiding over-fitting with large data sets.


Interventions for treating oral mucositis for patients with cancer receiving treatment.
Treatment of cancer is increasingly effective but associated with short and long term side effects. Oral side effects, including oral mucositis (mouth ulceration), remain a major source of illness despite the use of a variety of agents to treat them. To assess the effectiveness of interventions for treating oral mucositis or its associated pain in patients with cancer receiving chemotherapy or radiotherapy or both. Electronic searches of Cochrane Oral Health Group and PaPaS Trials Registers (to 1 June 2010), CENTRAL via The Cochrane Library (to Issue 2, 2010), MEDLINE via OVID (1950 to 1 June 2010), EMBASE via OVID (1980 to 1 June 2010), CINAHL via EBSCO (1980 to 1 June 2010), CANCERLIT via PubMed (1950 to 1 June 2010), OpenSIGLE (1980 to 1 June 2010) and LILACS via the Virtual Health Library (1980 to 1 June 2010) were undertaken. Reference lists from relevant articles were searched and the authors of eligible trials were contacted to identify trials and obtain additional information. All randomised controlled trials comparing agents prescribed to treat oral mucositis in people receiving chemotherapy or radiotherapy or both. Outcomes were oral mucositis, time to heal mucositis, oral pain, duration of pain control, dysphagia, systemic infection, amount of analgesia, length of hospitalisation, cost and quality of life. Data were independently extracted, in duplicate, by two review authors. Authors were contacted for details of randomisation, blindness and withdrawals. Risk of bias assessment was carried out on six domains. The Cochrane Collaboration statistical guidelines were followed and risk ratio (RR) values calculated using fixed-effect models (less than 3 trials in each meta-analysis). Thirty-two trials involving 1505 patients satisfied the inclusion criteria. Three comparisons for mucositis treatment including two or more trials were: benzydamine HCl versus placebo, sucralfate versus placebo and low level laser versus sham procedure. Only the low level laser showed a reduction in severe mucositis when compared with the sham procedure, RR 5.28 (95% confidence interval (CI) 2.30 to 12.13).Only 3 comparisons included more than one trial for pain control: patient controlled analgesia (PCA) compared to the continuous infusion method, therapist versus control, cognitive behaviour therapy versus control. There was no evidence of a difference in mean pain score between PCA and continuous infusion, however, less opiate was used per hour for PCA, mean difference 0.65 mg/hour (95% CI 0.09 to 1.20), and the duration of pain was less 1.9 days (95% CI 0.3 to 3.5). There is weak and unreliable evidence that low level laser treatment reduces the severity of the mucositis. Less opiate is used for PCA versus continuous infusion. Further, well designed, placebo or no treatment controlled trials assessing the effectiveness of interventions investigated in this review and new interventions for treating mucositis are needed.


Atrial fibrillation: overview of therapeutic trials.
Atrial fibrillation (AF) is the most common sustained cardiac arrhythmia treated by physicians, and a plethora of therapeutic trials deal with selected aspects of its management. This overview attempts to categorize and summarize the available studies. A key to management of AF is a clinical classification schema that provides a framework for application of the available treatment modalities. Such a classification is provided. Antithrombotic trials have demonstrated the remarkable efficacy of warfarin and more modest effect of acetylsalicylic acid for prevention of stroke; these data are summarized. Cardioversion to restore sinus rhythm is an important aspect of management of AF, particularly of persistent and new onset AF. In this review pharmacological cardioversion is emphasized. The data concerning the use of various drugs for pharmacological cardioversion are reviewed. Many, but not all, agents have been shown to have efficacy in this regard, but efficacy with drugs is lower than that with electrical cardioversion and, in the case of amiodarone, may be delayed. For recurrent AF, the two major rhythm management approaches are maintenance of rhythm and heart rate control. Trials of pharmacological and nonpharmacological therapies for these purposes are reviewed and summarized. Management of AF is an active area of research, and the present review is intended as a foundation upon which new information can be added.


Enhancing Caries Detection in Bitewing Radiographs Using YOLOv7.
The study aimed to evaluate the impact of image size, area of detection (IoU) thresholds and confidence thresholds on the performance of the YOLO models in the detection of dental caries in bitewing radiographs. A total of 2575 bitewing radiographs were annotated with seven classes according to the ICCMS™ radiographic scoring system. YOLOv3 and YOLOv7 models were employed with different configurations, and their performances were evaluated based on precision, recall, F1-score and mean average precision (mAP). Results showed that YOLOv7 with 640 × 640 pixel images exhibited significantly superior performance compared to YOLOv3 in terms of precision (0.557 vs. 0.268), F1-score (0.555 vs. 0.375) and mAP (0.562 vs. 0.458), while the recall was significantly lower (0.552 vs. 0.697). The following experiment found that the overall mAPs did not significantly differ between 640 × 640 pixel and 1280 × 1280 pixel images, for YOLOv7 with an IoU of 50% and a confidence threshold of 0.001 (p = 0.866). The last experiment revealed that the precision significantly increased from 0.570 to 0.593 for YOLOv7 with an IoU of 75% and a confidence threshold of 0.5, but the mean-recall significantly decreased and led to lower mAPs in both IoUs. In conclusion, YOLOv7 outperformed YOLOv3 in caries detection and increasing the image size did not enhance the model's performance. Elevating the IoU from 50% to 75% and confidence threshold from 0.001 to 0.5 led to a reduction of the model's performance, while simultaneously improving precision and reducing recall (minimizing false positives and negatives) for carious lesion detection in bitewing radiographs.


Holstein-Friesian strain and feed effects on milk production, body weight, and body condition score profiles in grazing dairy cows.
Data from 113 lactations across 76 cows between the years 2002 to 2004 were used to determine the effect of strain of Holstein-Friesian (HF) dairy cow and concentrate supplementation on milk production, body weight (BW), and body condition score (BCS; 1 to 5 scale) lactation profiles. New Zealand (NZ) and North American (NA) HF cows were randomly allocated to 1 of 3 levels of concentrate supplementation [0, 3, or 6 kg of dry matter (DM)/cow per d] on a basal pasture diet. The Wilmink exponential model was fitted within lactation (Y(DIM) = a + b e(-0.05 x DIM) + c x DIM). The median variation explained by the function for milk yield was 86%, between 62 and 69% for milk composition, and 80 and 70% for BW and BCS, respectively. North American cows and cows supplemented with concentrates had greater peak and 270-d milk yield. Concentrate supplementation tended to accelerate the rate of incline to peak milk yield, but persistency of lactation was not affected by either strain of HF or concentrate supplementation. No significant strain by diet interaction was found for parameters reported. New Zealand cows reached nadir BCS 14 d earlier and lost less BW (22 kg) postcalving than NA cows. Concentrate supplementation reduced the postpartum interval to nadir BW and BCS, and incrementally increased nadir BCS. New Zealand cows gained significantly more BCS (i.e., 0.9 x 10(-3) units/d more) postnadir than NA cows, and the rate of BCS replenishment increased linearly with concentrate supplementation from 0.5 x 10(-3) at 0 kg of DM/d to 0.8 x 10(-3) and 1.6 x 10(-3) units/d at 3 and 6 kg of DM/d concentrates, respectively. Although there was no significant strain by diet interaction for parameters reported, there was a tendency for a strain by diet interaction in 270-d BCS, suggesting that the effect of concentrate supplementation on BCS gain was, at least partly, strain dependent.


Season-specific and guild-specific effects of anthropogenic landscape modification on metacommunity structure of tropical bats.
Fragmentation per se due to human land conversion is a landscape-scale phenomenon. Accordingly, assessment of distributional patterns across a suite of potentially connected communities (i.e. metacommunity structure) is an appropriate approach for understanding the effects of landscape modification and complements the plethora of fragmentation studies that have focused on local community structure. To date, metacommunity structure within human-modified landscapes has been assessed with regard to nestedness along species richness gradients. This is problematic because there is little support that species richness gradients are associated with the factors moulding species distributions. More importantly, many alternative patterns are possible, and different patterns may manifest during different seasons and for different guilds because of variation in resource availability and resource requirements of taxa. We determined the best-fit metacommunity structure of a phyllostomid bat assemblage, frugivore ensemble, and gleaning animalivore ensemble within a human-modified landscape in the Caribbean lowlands of Costa Rica during the dry and wet seasons to elucidate important structuring mechanisms. Furthermore, we identified the landscape characteristics associated with the latent gradient underlying metacommunity structure. We discriminated among multiple metacommunity structures by assessing coherence, range turnover, and boundary clumping of an ordinated site-by-species matrix. We identified the landscape characteristics associated with the latent gradient underlying metacommunity structure via hierarchical partitioning. Metacommunity structure was never nested nor structured along a richness gradient. The phyllostomid assemblage and frugivore ensemble exhibited Gleasonian structure (range turnover along a common gradient) during the dry season and Clementsian structure (range turnover and shared boundaries along a common gradient) during the wet season. Distance between forest patches and forest edge density structured the phyllostomid metacommunity during the dry and wet seasons, respectively. Proportion of pasture and forest patch density structured the frugivore metacommunity during the dry season. Gleaning animalivores exhibited chequerboard structure (mutually exclusive species pairs) during the dry season and random structure during the wet season. Metacommunity structure was likely mediated by differential resource use or interspecific relationships. Furthermore, the interaction between landscape characteristics and seasonal variation in resources resulted in season-specific and guild-specific distributional patterns.


Exponentially tapered T-tube model in the characterization of arterial non-uniformity.
There is no question that geometric and elastic tapering is the rule of the nature of the mammalian arterial system. In this investigation, we determine the role of an exponentially tapered T-tube model to characterize the non-uniform nature and physical properties of the vasculature. The tapering feature could be inferred to show a connection between characteristic impedance at the distal of the tube and that at the entrance of the tube. Pulsatile pressure and flow signals of the ascending aorta were measured in 12 closed-chest, anesthetized dogs. This non-uniform T-tube model fits the measured aortic pressure waveform well. The mathematical and experimental model impedance spectra are similar. In the basal state, the distal characteristic impedance is 9.5% higher than the input characteristic impedance. The input characteristic impedance estimated in the model is compatible with that calculated by averaging high-frequency moduli of impedance data points obtained from the ratio of the corresponding harmonics of pressure and flow. Furthermore, the exponentially tapered T-tube model has a close estimate in wave transit time when compared with that computed by the impulse response of the filtered aortic input impedance. These data suggest that inclusion of tube tapering improves the mathematical model so that it is capable of characterizing the non-uniform nature of the arterial system. We conclude that the non-uniform properties of wave-transmission paths play an important role in governing the behavior of an asymmetric T-tube for the representation of the arterial system.


Relationships between radiographic change, pulmonary function, and bronchoalveolar lavage fluid lymphocytes in farmer's lung disease.
Ninety four dairy farmers were investigated by chest radiography, pulmonary function tests, and bronchoalveolar lavage. They were divided into five groups--1: 11 subjects with acute farmer's lung; 2: 25 subjects with previously diagnosed farmer's lung who had stayed on their farm; 3: 15 farmers with previously diagnosed farmer's lung who had left the farm; 4: 23 precipitin positive symptomless farmers; 5: 20 precipitin negative symptomless farmers. The study evaluated the relationships between radiographic changes measured with a scoring system derived from the International Labour Office (ILO) classification, the results of pulmonary function tests, and bronchoalveolar lavage fluid. Thirty eight subjects had radiographic evidence of interstitial pulmonary infiltrates. Group 1 had the highest percentage of lymphocytes recovered by bronchoalveolar lavage (mean 66.3 (SD 19.2]. For all subjects carbon monoxide transfer factor (TLCO) and total lung capacity were negatively correlated with radiographic changes (r = -0.45 and -0.30; p less than 0.001 and less than 0.01 respectively). TLCO was also negatively correlated with radiographic change in group 2 (r = -0.59, p less than 0.005). The percentage of lavage lymphocytes was correlated with radiographic changes for all subjects (r = 0.36, p less than 0.001), but this correlation was not seen within groups. This study shows good correlation between radiographic abnormalities, pulmonary function changes and the cellular composition of bronchoalveolar lavage fluid.


Presumed residual thymic tissue is a common finding in thoracic computed tomography in adult dogs.
Residual thymic tissue is a common incidental finding in thoracic CT of human adults. To determine whether presumed residual thymic tissue is also a common incidental finding in adult dogs, a two part-study was performed. The first part was a prospective, descriptive design where CT examination was performed in six canine cadavers within 24 h after death and presumed residual thymic tissue was examined pathologically. The second part of the study was a retrospective, analytical design where medical records of our institution were searched for thoracic CT scans of adult dogs performed in the year 2020. Age, sex, breed, presence of presumed thymic tissue, location, shape, attenuation, homogeneity, and width of the tissue were recorded and comparisons were performed using these data. In 4 of 6 of the prospective cases, thymic tissue was present on histology and in 2 of 6 dogs the presence of thymic tissue could not be confirmed. For the retrospective study, in 161/169 (95.3%) cases with presumed residual thymic tissue were detected. Shape and size were highly variable with either homogeneous (46.6%), heterogeneous (42.9%), or mixed (10.6%) attenuation. Dogs with presumed residual thymic tissue were significantly younger (median: 9.1 years; range: 1.2-14.3 vs. median: 10.5 years; range: 9.4-12.3) as were dogs with homogeneous attenuation of the tissue (median: 8.1 years; range: 1.2-14.3 vs. median: 9.5 years; range: 4.0-14.3). In conclusion, results indicated that presumed residual thymic tissue is a common CT finding in adult dogs and can be considered incidental.


Renal Impairment Hampers Bisphosphonate Treatment in a Quarter of Lung Cancer Patients with Bone Metastasis.
Renal function impairment in lung cancer patients with bone metastases was investigated, as this can limit the application of bisphosphonates representing the gold standard in the management of such cases. Clinicopathological data of 570 lung cancer patients were retrospectively analysed for changes in renal function parameters. Comorbidities included hypertension (50%), COPD (33%) and diabetes mellitus (15%). Statistical analysis was performed with Fisher's exact tests and a Cox proportional hazards model. In patients suffering from hypertension, both median serum creatinine and blood urea nitrogen (BUN) were higher (81.9 versus 75.8 μmol/L, p < 0.001 and 6.0 versus 5.7 mmol/L, p = 0.005, respectively). Such a difference could not be observed in patients with diabetes. In patients with COPD, only serum creatinine was higher (81.1 versus 77.3 μmol/L, p = 0.004). In the whole cohort, we found that while at the time of lung cancer diagnosis the ratio of patients in the pathological range (PRR) was 8.67% for serum creatinine (median: 75 μmol/L) and 14.16% for BUN (median: 5.4 mmol/L), at the time of bone metastasis the PRR for serum creatinine increased to 16.11% (median: 77.0 μmol/L) and for BUN to 24.07% (median: 6.0 mmol/L), which is a significant increase for both parameters (p < 0.001). For the whole cohort, the last laboratory results showed a 26.37% PRR for serum creatinine and 45.66% PRR for BUN (significant increase for both, p < 0.001). Multivariate analysis revealed that patients with hypertension had a higher chance for switching to the pathological range sooner (p = 0.033, HR: 1.372, CI: 1.025-1.835). Also, the appearance of the bone metastasis correlated with an acceleration of the onset of such a switch (p < 0.001, HR: 2.655, CI: 1.581-4.456). Our results suggest that renal function is impaired in a significant proportion of patients with lung cancer and highlight the importance of non-nephrotoxic drug in the management of bone metastases.


MP4CO, a pegylated hemoglobin saturated with carbon monoxide, is a modulator of HO-1, inflammation, and vaso-occlusion in transgenic sickle mice.
Transgenic sickle mice expressing β(S) hemoglobin have activated vascular endothelium in multiple organs that exhibits enhanced expression of NF-ĸB and adhesion molecules and promotes microvascular stasis in sickle, but not normal, mice in response to hypoxia/reoxygenation (H/R), or heme. Induction of heme oxygenase-1 (HO-1) or administration of its products, carbon monoxide (CO) or biliverdin, inhibits microvascular stasis in sickle mice. Infusion of human hemoglobin conjugated with polyethylene glycol and saturated with CO (MP4CO) markedly induced hepatic HO-1 activity and inhibited NF-ĸB activation and H/R-induced microvascular stasis in sickle mice. These effects were mediated by CO; saline or MP4 saturated with O2 (MP4OX) had little to no effect on H/R-induced stasis, though unmodified oxyhemoglobin exacerbated stasis. The HO-1 inhibitor, tin protoporphyrin, blocked MP4CO protection, consistent with HO-1 involvement in the protection afforded by MP4CO. MP4CO also induced nuclear factor-erythroid 2 p45-related factor 2 (Nrf2), an important transcriptional regulator of HO-1 and other antioxidant genes. In a heterozygous (hemoglobin-AS) sickle mouse model, intravenous hemin induced cardiovascular collapse and mortality within 120 minutes, which was significantly reduced by MP4CO, but not MP4OX. These data demonstrate that MP4CO induces cytoprotective Nrf2 and HO-1 and decreases NF-ĸB activation, microvascular stasis, and mortality in transgenic sickle mouse models.


Correlation of anthropometric measurements, strength, anterior cruciate ligament size, and intercondylar notch characteristics to sex differences in anterior cruciate ligament tear rates.
We performed a prospective study based on the hypothesis that physiologic differences exist between men and women in strength after adjustments for body weight; that the size of the anterior cruciate ligament is proportionate to the strength of its antagonists, the quadriceps muscles; and that women have a relatively small anterior cruciate ligament, thus predisposing them to a disproportionate number of anterior cruciate ligament injuries. One hundred matched high school basketball players, 50 male and 50 female, were evaluated with anthropometric measurements, body fat analysis, muscle strength evaluation, and magnetic resonance imaging measurements of the intercondylar notch and cross-sectional area of the anterior cruciate ligament at the outlet. The male players were taller and heavier than their female counterparts, although they had 11% less body fat. Male players had statistically greater quadriceps and hamstring muscle strength than female players, even when adjustments were made for body weight. With adjustments for body weight, the size of the anterior cruciate ligament in girls was found to be statistically smaller than in boys. There was no statistically significant difference in the notch width index between the sexes. The study data support our hypothesis that sex differences in anterior cruciate ligament tear rates are caused primarily by several interrelated intrinsic factors. Most importantly, stiffness and muscular strength increase stress on the anterior cruciate ligament in female athletes. The anterior cruciate ligament, when adjustments have been made for body weight, is smaller in female athletes, and therefore, probably does not compensate for the lack of stiffness and strength.


Involvement of the JNK/FOXO3a/Bim Pathway in Neuronal Apoptosis after Hypoxic-Ischemic Brain Damage in Neonatal Rats.
c-Jun N-terminal kinase (JNK) plays a key role in the regulation of neuronal apoptosis. Previous studies have revealed that forkhead transcription factor (FOXO3a) is a critical effector of JNK-mediated tumor suppression. However, it is not clear whether the JNK/FOXO3a pathway is involved in neuronal apoptosis in the developing rat brain after hypoxia-ischemia (HI). In this study, we generated an HI model using postnatal day 7 rats. Fluorescence immunolabeling and Western blot assays were used to detect the distribution and expression of total and phosphorylated JNK and FOXO3a and the pro-apoptotic proteins Bim and CC3. We found that JNK phosphorylation was accompanied by FOXO3a dephosphorylation, which induced FOXO3a translocation into the nucleus, resulting in the upregulation of levels of Bim and CC3 proteins. Furthermore, we found that JNK inhibition by AS601245, a specific JNK inhibitor, significantly increased FOXO3a phosphorylation, which attenuated FOXO3a translocation into the nucleus after HI. Moreover, JNK inhibition downregulated levels of Bim and CC3 proteins, attenuated neuronal apoptosis and reduced brain infarct volume in the developing rat brain. Our findings suggest that the JNK/FOXO3a/Bim pathway is involved in neuronal apoptosis in the developing rat brain after HI. Agents targeting JNK may offer promise for rescuing neurons from HI-induced damage.


Effects of laparotomy on spontaneous exploratory activity and conditioned operant responding in the rat: a model for postoperative pain.
Treatment of postsurgical pain is a major use of analgesics, particularly after abdominal surgery. Analgesics display a number of limiting side effects, including sedation, cognitive impairment, and ileus. Although several postoperative rodent models have been developed, these models do not address these concerns. A model is presented in the rat in which a subcostal incision is performed, penetrating into the peritoneal cavity. The behavioral effects of this surgical procedure are assessed using exploratory locomotor activity and conditioned operant responding. The effects of morphine and ketorolac were assessed in both behavioral paradigms. Laparotomy decreased ambulation and rearing by approximately 50% 24 h after surgery, and stereotypy (small confined movements) was affected to a lesser degree. The effects of laparotomy on conditioned operant responding were more complex. Total number of sucrose pellets earned was decreased for 2-3 days after laparotomy; however, the amount of time required was increased for up to 2 weeks. Morphine reversed the effects of surgery on ambulation and stereotypy but not rearing, and the dose-effect curve for morphine was shifted to the left by 5 mg/kg ketorolac. Ketorolac produced significant improvement in operant responding after laparotomy, and coadministration of ineffective doses of morphine and ketorolac produced a positive response. The current model is consistent with behavioral aspects of postoperative pain seen clinically. The effects of morphine and ketorolac alone and in combination were consistent with the reported analgesic efficacy and occurrence of side effects found with these agents clinically.


Analysis of heart development in cultured rat embryos.
The long-range goal of this research is to establish an in vitro system that will permit pertubation of mammalian heart development and in situ examination of the cellular and molecular events underlying cardiac morphogenesis. Rat embryos at 9.5-11.5 days of gestation were placed in culture bottles containing rat serum and Tyrode's solution. Embryos cultured for 24 and 48 h were compared to age-matched in vivo controls for morphological score, morphometric analysis of heart development, and confocal and electron microscopic analysis of myofiber pattern formation. Morphological scores indicated that embryos cultured for 24 h from day 9.5 to 10.5 had essentially normal development when compared to age-matched embryos allowed to develop in vivo. Development of embryos maintained for 48 h in culture was slightly delayed at 66-68% of age matched in vivo embryos. Analysis of hearts from embryos allowed to develop 9.5-11.5 days in vivo plus 24 and 48 h in culture showed that the ventricular thickness and height, as well as the truncal, atrial and ventricular diameters were equivalent to those of hearts from age-matched in vivo controls. Hearts from embryos allowed to develop from 11.5-12.5 days in vitro and cultured for 24 and 48 h had smaller left ventricular and atrial dimensions than controls. Cardiac myofibrillogenesis and myofibrillar pattern formation in embryos cultured from 9.5 days of in vivo development for 48 h were also normal. These studies indicate that the rat whole embryo culture system is a useful model to study several critical periods in mammalian heart development.


Genetic analysis of the relationships between the cell surface and the nuclei in Paramecium tetraurella.
In Paramecium tetraurelia, a number of mutations have been shown to affect simultaneously cortical organization (attachment of trichocysts to the cortex) and nuclear divisions (Ruiz et al. 1976). In order to analyze the genetic and physiological basis of this correlation, and we have isolated new mutations affecting the properties of the trichocysts and studied their genetic relationships with other previously known mutations. Of 24 to 28 loci controlling the biogenesis and properties of the trichocysts, mutations only in the 16 to 20 loci tht control trichocysts attachment to the cortex result in nuclear defects. Cytological observations show that all these mutants display the same set of nucler abnormalities: in particular, rounded shape of the resting macronucleus mispositioning and defective elongation of the dividing macronucleus and unequal repartition of the marco- and micronuclei. This common syndrome is independent of both the mutagenic origin and the mutated locus. Furthermore, by microinjection, it is possible to localize the site of action of the mutations in either the trichocyst compartment or the nontrichocyst compartment. It was found by this technique that the nuclear syndrome is also independent of the site of action of the mutation. All the genetic and physiological data support the conclusion that the nuclear defects are the consequence of the lack of trichocyst attachement to the cortex: in wild-type cells, trichocyst attachment would induce a membranar or perimembranar state necessary for correct nuclear positioning during cell division. In the absence of trichocyst attachment, the cortical control of nuclear division would be abolished. The possible involvement of cytoskeletal links between surface and nuclei is discussed.


Differential protein expression in the hippocampi of resilient individuals identified by digital spatial profiling.
Clinical symptoms correlate with underlying neurodegenerative changes in the vast majority of people. However, an intriguing group of individuals demonstrate neuropathologic changes consistent with Alzheimer disease (AD) yet remain cognitively normal (termed "resilient"). Previous studies have reported less overall neuronal loss, less gliosis, and fewer comorbidities in these individuals. Herein, NanoString GeoMx™ Digital Spatial Profiler (DSP) technology was utilized to investigate protein expression differences comparing individuals with dementia and AD neuropathologic change to resilient individuals. DSP allows for spatial analysis of protein expression in multiple regions of interest (ROIs) on formalin-fixed paraffin-embedded sections. ROIs in this analysis were hippocampal neurofibrillary tangle (NFT)-bearing neurons, non-NFT-bearing neurons, and their immediate neuronal microenvironments. Analyses of 86 proteins associated with CNS cell-typing or known neurodegenerative changes in 168 ROIs from 14 individuals identified 11 proteins displaying differential expression in NFT-bearing neurons of the resilient when compared to the demented (including APP, IDH1, CD68, GFAP, SYP and Histone H3). In addition, IDH1, CD68, and SYP were differentially expressed in the environment of NFT-bearing neurons when comparing resilient to demented. IDH1 (which is upregulated under energetic and oxidative stress) and PINK1 (which is upregulated in response to mitochondrial dysfunction and oxidative stress) both displayed lower expression in the environment of NFT-bearing neurons in the resilient. Therefore, the resilient display less evidence of energetic and oxidative stress. Synaptophysin (SYP) was increased in the resilient, which likely indicates better maintenance of synapses and synaptic connections. Furthermore, neurofilament light chain (NEFL) and ubiquitin c-terminal hydrolase (Park5) were higher in the resilient in the environment of NFTs. These differences all suggest healthier intact axons, dendrites and synapses in the resilient. In conclusion, resilient individuals display protein expression patterns suggestive of an environment containing less energetic and oxidative stress, which in turn results in maintenance of neurons and their synaptic connections.


Chrysanthemum ethanol extract induced loss of Kupffer cells via the mitochondria-dependent apoptotic pathway.
Chrysanthemum has been viewed as an important traditional Chinese medicine (TCM) with a long history. Research studies indicated many potential pharmaceutical effects of chrysanthemum extract. However, hardly any investigation has been performed to describe its toxicity. In this study, acute application of chrysanthemum ethanol extract (CEE, 300 mg kg-1) was found to induce apoptosis of hepatic Kupffer cells in vivo. CEE was also observed to induce apoptosis of RAW264.7 cells in a dose- and time-dependent manner. Further analysis using flow cytometry and western blotting revealed that CEE induced apoptosis of RAW264.7 cells via a mitochondria-dependent pathway. After a HPLC combined screening assay, we narrowed down the toxicity caused by the petroleum extract of CEE (CEE-PE, 66 μg mL-1). In vivo effects of CEE-PE were also tested in mice. Additionally, nine potential toxic compounds were isolated and identified from CEE-PE. In all, we found that components with small polarities in CEE could induce apoptosis of Kupffer cells and macrophages via a mitochondrial dependent pathway, which might draw attention to the safety issues of everyday use of chrysanthemum.


Novel Large Sulfur Bacteria in the Metagenomes of Groundwater-Fed Chemosynthetic Microbial Mats in the Lake Huron Basin.
Little is known about large sulfur bacteria (LSB) that inhabit sulfidic groundwater seeps in large lakes. To examine how geochemically relevant microbial metabolisms are partitioned among community members, we conducted metagenomic analysis of a chemosynthetic microbial mat in the Isolated Sinkhole, which is in a deep, aphotic environment of Lake Huron. For comparison, we also analyzed a white mat in an artesian fountain that is fed by groundwater similar to Isolated Sinkhole, but that sits in shallow water and is exposed to sunlight. De novo assembly and binning of metagenomic data from these two communities yielded near complete genomes and revealed representatives of two families of LSB. The Isolated Sinkhole community was dominated by novel members of the Beggiatoaceae that are phylogenetically intermediate between known freshwater and marine groups. Several of these Beggiatoaceae had 16S rRNA genes that contained introns previously observed only in marine taxa. The Alpena fountain was dominated by populations closely related to Thiothrix lacustris and an SM1 euryarchaeon known to live symbiotically with Thiothrix spp. The SM1 genomic bin contained evidence of H2-based lithoautotrophy. Genomic bins of both the Thiothrix and Beggiatoaceae contained genes for sulfur oxidation via the rDsr pathway, H2 oxidation via Ni-Fe hydrogenases, and the use of O2 and nitrate as electron acceptors. Mats at both sites also contained Deltaproteobacteria with genes for dissimilatory sulfate reduction (sat, apr, and dsr) and hydrogen oxidation (Ni-Fe hydrogenases). Overall, the microbial mats at the two sites held low-diversity microbial communities, displayed evidence of coupled sulfur cycling, and did not differ largely in their metabolic potentials, despite the environmental differences. These results show that groundwater-fed communities in an artesian fountain and in submerged sinkholes of Lake Huron are a rich source of novel LSB, associated heterotrophic and sulfate-reducing bacteria, and archaea.


Pellet-Fed Gasifier Stoves Approach Gas-Stove Like Performance during in-Home Use in Rwanda.
Nearly all households in Rwanda burn solid fuels for cooking. A private firm in Rwanda is distributing forced-draft pellet-fed semigasifier cookstoves and fuel pellets. We measured in-use emissions of pollutants including fine particulate matter (PM2.5), organic and elemental carbon (OC, EC), black carbon (BC), and carbon monoxide (CO) in 91 uncontrolled cooking tests (UCTs) of both pellet and baseline (wood; charcoal) stoves. We observed >90% reductions in most pollutant emission factors/rates from pellet stoves compared to baseline stoves. Pellet stoves performed far better than gasifier stoves burning unprocessed wood, and consistent with ISO tiers 4 and 5 for PM2.5 and CO, respectively. Pellet stoves were generally clean, but performance varied; emissions from the dirtiest pellet tests matched those from the cleanest traditional stove tests. Our real-time data suggest that events occurring during ignition and the end of testing (e.g., refueling, char burnout) drive high emissions during pellet tests. We use our data to estimate potential health and climate cobenefits from stove adoption. This analysis suggests that pellet stoves have the potential to provide health benefits far above previously tested biomass stoves and approaching modern fuel stoves (e.g., LPG). Net climate impacts of pellet stoves range from similar to LPG to negligible, depending on biomass source and upstream emissions.


Unobtrusive Sleep Position Classification Using a Novel Optical Tactile Sensor.
Unobtrusive sleep position classification is essential for sleep monitoring and closed-loop intervention systems that initiate position changes. In this paper, we present a novel unobtrusive under-mattress optical tactile sensor for sleep position classification. The sensor uses a camera to track particles embedded in a soft silicone layer, inferring the deformation of the silicone and therefore providing information about the pressure and shear distributions applied to its surface.We characterized the sensitivity of the sensor after placing it under a conventional mattress and applying different weights (258 g, 500 g, 5000 g) on top of the mattress in various predefined locations. Moreover, we collected multiple recordings from a person lying in supine, lateral left, lateral right, and prone positions. As a proof-of-concept, we trained a neural network based on convolutional layers and residual blocks that classified the lying positions based on the images from the tactile sensor.We observed a high sensitivity of the optical tactile sensor: Even after placing the sensor below a conventional mattress, we were able to detect our lowest test weight of 258 g. Using the neural network, we were able to classify the four sleep positions, lateral left, lateral right, prone, and supine with a classification accuracy of 91.2 %.The high sensitivity of the sensor, as well as the good performance in the classification task, demonstrate the feasibility of using such a sensor in a robotic bed setup.Clinical Relevance- Positional Obstructive Sleep Apnea is highly prevalent across the general population. Today's gold standard treatment of using CPAP ventilation is often not accepted, leading to unwanted treatment cessations. Alternative treatments, such as positional interventions through robotic beds are highly promising. However, these beds require reliable detection of the lying position. In this paper, we present a novel, scalable and completely unobtrusive sensor that is concealed under the mattress while classifying sleeping positions with high accuracy.


Patient centered assessment of ankylosing spondylitis-specific health related quality of life: evaluation of the Patient Generated Index.
To evaluate the Patient Generated Index (PGI) for the measurement of individualized health related quality of life in ankylosing spondylitis (AS). The PGI asks patients to nominate areas of their lives affected by their disease that they consider the most important. Semistructured interviews with AS patients produced a trigger list of areas of life affected by AS. The PGI was self-completed by UK patients taking part in a multicenter postal survey. The instrument was assessed for data quality, reliability, validity, and responsiveness. The PGI had acceptable completion rates. Scores covered the available range and approximated normality. Test-retest reliability estimates support the use of the PGI in group evaluation (intraclass correlation coefficient > 0.80). Comparisons with scores for other health status instruments provided evidence for the validity of the PGI. The largest levels of correlation were found for the AS Quality of Life Questionnaire (ASQoL) and the EuroQol. The informed and open format of the PGI had the strongest linear relationship with responses to both specific and general health transition questions (p < 0.01), and was the most responsive format of the PGI. The PGI is the first AS-specific individualized measure of health related quality of life. There is good support for the content validity of the instrument and patient acceptability is high. Adequate levels of data quality and reliability support the use of the PGI in group evaluation. Moderate levels of responsiveness to changes in health were produced by the informed and open format of the PGI.


Quantitation of apo-, mono-, and diferric transferrin by polyacrylamide gradient gel electrophoresis in patients with disorders of iron metabolism.
We have developed a polyacrylamide gradient gel electrophoretic method to quantitate apo-, mono-, and diferric transferrin based upon differences in their molecular size. Purified transferrin saturated to different extents (3% to 98%) with iron showed proportions of the three forms as predicted from an approximately random distribution of iron between the two metal-binding sites. The iron distributions in sera of 14 normal individuals similarly correlated with the predicted values. In contrast, 22 of 43 patients with diseases associated with abnormalities in iron or transferrin metabolism had a disproportionate increase in monoferric transferrin. This abnormality occurred in seven of nine patients who had received bone marrow transplants, seven of 14 with chronic liver disease, and eight of nine menstruating women with probable iron deficiency anemia. Interestingly, 11 patients with malabsorption or chronic renal disease had normal iron distributions. The finding of abnormal distributions of iron on transferrin suggests that gradient gel analysis may be a useful tool for studying the physiologic mechanisms controlling iron utilization.


Adenosine receptor activation induces vascular endothelial growth factor in human retinal endothelial cells.
Adenosine, released in increased amounts by hypoxic tissues, is thought to be an angiogenic factor that links altered cellular metabolism caused by oxygen deprivation to compensatory angiogenesis. Adenosine interacts with 4 subtypes of G protein-coupled receptors, termed A(1), A(2A), A(2B), and A(3). We investigated whether adenosine causes proliferation of human retinal endothelial cells (HRECs) and synthesis of vascular endothelial growth factor (VEGF) and, if so, which adenosine receptor subtype mediates these effects. The nonselective adenosine receptor agonist 5'-N-ethylcarboxamidoadenosine (NECA), in a concentration-dependent manner, increased both VEGF mRNA and protein expression by HRECs, as well as proliferation. This proliferative effect of NECA was inhibited by the addition of anti-human VEGF antibody. NECA also increased insulin-like growth factor-I and basic fibroblast growth factor mRNA expression in a time-dependent manner and cAMP accumulation in these cells. In contrast, neither the A(1) agonist N(6)-cyclopentyladenosine nor the A(2A) agonist 2-p-(2-carboxyethyl) phenethylamino-NECA caused any of the above effects of NECA. The effects of NECA were not significantly attenuated by either the A(2A) antagonist SCH58261 or the A(1) antagonist 8-cyclopentyl-1, 3-dipropylxanthine. However, the nonselective adenosine receptor antagonist xanthine amine congener completely inhibited the effects of NECA. Addition of antisense oligonucleotide complementary to A(2B) adenosine receptor mRNA inhibited VEGF protein production by HRECs after NECA stimulation. Thus, the A(2B) adenosine receptor subtype appears to mediate the actions of adenosine to increase growth factor production, cAMP content, and cell proliferation of HRECs. Adenosine activates the A(2B) adenosine receptor in HRECs, which may lead to neovascularization by a mechanism involving increased angiogenic growth factor expression.


The Rat Genome Database pathway portal.
The set of interacting molecules collectively referred to as a pathway or network represents a fundamental structural unit, the building block of the larger, highly integrated networks of biological systems. The scientific community's interest in understanding the fine details of how pathways work, communicate with each other and synergize, and how alterations in one or several pathways may converge into a disease phenotype, places heightened demands on pathway data and information providers. To meet such demands, the Rat Genome Database [(RGD) http://rgd.mcw.edu] has adopted a multitiered approach to pathway data acquisition and presentation. Resources and tools are continuously added or expanded to offer more comprehensive pathway data sets as well as enhanced pathway data manipulation, exploration and visualization capabilities. At RGD, users can easily identify genes in pathways, see how pathways relate to each other and visualize pathways in a dynamic and integrated manner. They can access these and other components from several entry points and effortlessly navigate between them and they can download the data of interest. The Pathway Portal resources at RGD are presented, and future directions are discussed. Database URL: http://rgd.mcw.edu.


The dyadic nature of social information processing in boys' reactive and proactive aggression.
The correlation between boys' social cognitions and their aggressive behavior toward peers was examined as being actor driven, partner driven, or dyadic relationship driven. Eleven groups of 6 familiar boys each (N = 165 dyads) met for 5 consecutive days to participate in play sessions and social-cognitive interviews. With a variance partitioning procedure, boys' social-cognitive processes were found to vary reliably across their dyadic relationships. Furthermore, mixed models regression analyses indicated that hostile attributional biases toward a particular peer were related to directly observed reactive aggression toward that peer even after controlling for actor and partner effects, suggesting that these phenomena are dyadic or relationship oriented. On the other hand, the relation between outcome expectancies for aggression and the display of proactive aggression appeared to be more actor driven and partner driven that dyadic.


Distribution of selenoproteins in mouse mammary epithelial cells in vitro and in vivo.
Resolution of selenium-containing proteins synthesized by mouse mammary gland cells was achieved using the technique of two-dimensional gel electrophoresis. Radioactive selenium as H2[75Se]O3 was incorporated into relatively few proteins within mammary gland cells maintained in vitro and cells of mammary gland tissue in vivo. The pattern of selenoproteins obtained was identical qualitatively between a nontumorigenic differentiated cell line, COMMA-D, and a tumorigenic cell line, MOD. Eleven selenoproteins ranging in molecular weight from 12,000-78,000 were detected and a total of 25 spots were visible indicating charge heterogeneity of some of the proteins. A major selenoprotein (Mr 26,000) migrated identically with the subunit form of glutathione peroxidase, a well-characterized protein containing four selenocysteine residues. Other major selenoproteins had molecular weights of 58,000, 22,000, 18,000, and 14,000. Analysis of the total cellular protein extract and of each of the five major proteins indicated that selenium was incorporated as selenocysteine in the proteins. Incorporation of selenium as selenomethionine into cellular proteins was detected only when selenomethionine was provided in the culture medium. Cleavage of 75Se-labeled proteins with N-chlorosuccinimide produced polypeptides of different molecular weights indicating that the Mr 58,000, 26,000, and 22,000 selenoproteins were dissimilar in the amino acid sequences containing the selenoamino acid. The pattern of selenoproteins of mammary gland cells in vivo was similar to that obtained for cells in culture and most other tissues in vivo. These results provide evidence for the presence of multiple selenium-containing proteins in mammary epithelial cells. The possible significance of these proteins in selenium-mediated inhibition of cell growth awaits future clarification.


Nutrition and cancer: physiological interrelationships.
There are many tumors that have paraneoplastic syndromes. Furthermore, location of certain tumors can result in very specific effects on the host, especially tumors in the hypothalamus, the intestinal tract, or the liver. Finally, tumors of the immune system can have significant distant consequences. However, from direct experimental evidence, from model systems, and from the utilization of nutritional manipulation in the treatment of cancer, the data suggest very strongly that there is no unique cancer malnutrition. Early diagnosed cancer does not show malnutrition as a presenting symptom. Furthermore, all metabolic disturbances can be explained on the basis of the metabolic differences of tumor cells and normal cells and are very frequently proportional to the bulk of the tumor. The cachexia that is associated with malignancies is more likely cachexia in cancer patients than it is a specific cancer cachexia, unless the tumor burden is very large. This point was clearly made in a short review of the causes of cachexia in nearly 1500 cancer patients in Russia (145). Brennan also feels that most cases of malnutrition are uncomplicated starvation, and cancer cachexia has many features seen in major injury or sepsis (16). This distinction has great implications in the management of cancer patients.


Creatine nitrate supplementation strengthens energy status and delays glycolysis of broiler muscle via inhibition of LKB1/AMPK pathway.
This study aimed to evaluate the effects of dietary creatine nitrate (CrN) on growth performance, meat quality, energy status, glycolysis, and related gene expression of liver kinase B1/AMP-activated protein kinase (LKB1/AMPK) pathway in Pectoralis major (PM) muscle of broilers. A total of 240 male Arbor Acres broilers (28-day-old) were randomly allocated to one of 5 dietary treatments: the basal diet (control group), and the basal diets supplemented with 600 mg/kg guanidinoacetic acid (GAA), 300, 600, or 900 mg/kg CrN (identified as GAA600, CrN300, CrN600, or CrN900, respectively). We found that dietary GAA and CrN supplementation for 14 d from d 28 to 42 did not affect broiler growth performance, carcass traits, and textural characteristics of breast muscle. GAA600, CrN600, and CrN900 treatments increased pH24h and decreased drip loss of PM muscle compared with the control (P < 0.05). The PM muscles of CrN600 and CrN900 groups showed higher glycogen concentration and lower lactic acid concentration accompanied by lower activities of phosphofructokinase (PFK), pyruvate kinase (PK), and lactate dehydrogenase (LDH) (P < 0.05). Simultaneously, GAA600 and all CrN treatments increased concentration of muscle creatine, phosphocreatine (PCr) and ATP, and decreased AMP concentration and AMP/ATP ratio (P < 0.05). Meanwhile, the concentrations of muscle creatine, PCr, and ATP were increased linearly, while muscle AMP concentration and AMP/ATP ratio were decreased linearly and quadratic as the dose of CrN increased (P < 0.05). GAA600, CrN600, and CrN900 treatments upregulated mRNA expression of CreaT in PM muscle, and CrN600 and CrN900 treatments downregulated GAMT expression in liver and PM muscle compared with the control or GAA600 groups (P < 0.05). The mRNA expression of muscle LKB1, AMPKα1, and AMPKα2 was downregulated linearly in response to the increasing CrN level (P < 0.05). Overall, CrN showed better efficacy on strengthening muscle energy status and improve meat quality than GAA at the some dose. These results indicate that CrN may be a potential replacement for GAA as a new creatine supplement.


Stigma in Male Depression and Suicide: A Canadian Sex Comparison Study.
Stigma in men's depression and suicide can restrict help-seeking, reduce treatment compliance and deter individuals from confiding in friends and family. In this article we report sex comparison findings from a national survey of English-speaking adult Canadians about stigmatized beliefs concerning male depression and suicide. Among respondents without direct experience of depression or suicide (n = 541) more than a third endorsed the view that men with depression are unpredictable. Overall, a greater proportion of males endorsed stigmatizing views about male depression compared to female respondents. A greater proportion of female respondents endorsed items indicating that men who suicide are disconnected, lost and lonely. Male and female respondents with direct personal experience of depression or suicide (n = 360) strongly endorsed stigmatizing attitudes toward themselves and a greater proportion of male respondents indicated that they would be embarrassed about seeking help for depression.


Automated Identification and Quantification of Signals in Multichannel Immunofluorescence Images: The SignalFinder-IF Platform.
Multimarker fluorescence analysis of tissue specimens offers the opportunity to probe the expression levels and locations of multiple markers in a single sample. Software is needed to fully capitalize on the advantages of this technology for sensitive, quantitative, and multiplexed data collection. A major challenge has been the automated identification and quantification of signals. We report on the software SignalFinder-IF, which meets that need. SignalFinder-IF uses a newly developed algorithm called Segment-Fit Thresholding, which showed robust performance for automated signal identification in side-by-side comparisons with several current methods. Two utilities provided with SignalFinder-IF enable downstream analyses. The first allows the quantification and mapping of relationships between an unlimited number of markers through user-defined sequences of AND, OR, and NOT operators. The second produces composite pictures of the signals or colocalization analysis on brightfield hematoxylin and eosin images, which is useful for understanding the morphologies and locations of cells meeting specific marker criteria. SignalFinder-IF enables high-throughput, rigorous analyses of whole-slide, multimarker data, and it promises to open new possibilities in many research and clinical applications.


[On the problem of microscopic diagnostics of death by drowning].
The objective of the present work was to estimate the results of microscopic investigations of organs and tissues for the purpose of forensic histological, forensic medical, micrologic, etc. expertise of the death by drowning in fresh water. A total of 3527 corpses of men and women aged from 3 to 77 years drowned in the Republic of Udmurtia in 1995-2011 were available for examination. The author demonstrates the possibility of using forensic medical investigations for the development of classification, elucidation of conditions and type of drowning. A reliable analysis of micrological data implies the necessity of their combination which is impossible in the framework of a single medical discipline; it is rather the interdisciplinary problem. The results of this study may contribute to the accumulation of the materials needed to create the working database.


Enhanced use of phylogenetic data to inform public health approaches to HIV among men who have sex with men.
The multidimensional nature and continued evolution of HIV epidemics among men who have sex with men (MSM) requires innovative intervention approaches. Strategies are needed that recognise the individual, social and structural factors driving HIV transmission; that can pinpoint networks with heightened transmission risk; and that can help target intervention in real time. HIV phylogenetics is a rapidly evolving field with strong promise for informing innovative responses to the HIV epidemic among MSM. Currently, HIV phylogenetic insights are providing new understandings of characteristics of HIV epidemics involving MSM, social networks influencing transmission, characteristics of HIV transmission clusters involving MSM, targets for antiretroviral and other prevention strategies and dynamics of emergent epidemics. Maximising the potential of HIV phylogenetics for HIV responses among MSM will require attention to key methodological challenges and ethical considerations, as well as resolving key implementation and scientific questions. Enhanced and integrated use of HIV surveillance, sociobehavioural and phylogenetic data resources are becoming increasingly critical for informing public health approaches to HIV among MSM.


Tamponade as the clinical onset of a cardiac angiosarcoma.
Primary tumors of the heart are usually benign, with the malignant forms being rare. In this article we report the case of an angiosarcoma with the only manifestation being a cardiac tamponade. When the patient being admitted to the hospital, an important differential diagnosis was pulmonary tumor. A mass in the right atrium was later detected in the echocardiogram. Further image studies, such as MRI, showed spectacular images of this mass suggestive of neoplasm. An intra-operatory biopsy diagnosed the mass as an angiosarcoma. Since there was no evidence of primary neoplasm in other studies carried out, the neoformation was diagnosed as a primary angiosarcoma. Surgery was carried out with a survival of only 13 months after the procedure. We also use this case to make a brief review of the primary tumors of the heart, especially angiosarcomas.


Quantitative 4D analyses of epithelial folding during Drosophila gastrulation.
Understanding the cellular and mechanical processes that underlie the shape changes of individual cells and their collective behaviors in a tissue during dynamic and complex morphogenetic events is currently one of the major frontiers in developmental biology. The advent of high-speed time-lapse microscopy and its use in monitoring the cellular events in fluorescently labeled developing organisms demonstrate tremendous promise in establishing detailed descriptions of these events and could potentially provide a foundation for subsequent hypothesis-driven research strategies. However, obtaining quantitative measurements of dynamic shapes and behaviors of cells and tissues in a rapidly developing metazoan embryo using time-lapse 3D microscopy remains technically challenging, with the main hurdle being the shortage of robust imaging processing and analysis tools. We have developed EDGE4D, a software tool for segmenting and tracking membrane-labeled cells using multi-photon microscopy data. Our results demonstrate that EDGE4D enables quantification of the dynamics of cell shape changes, cell interfaces and neighbor relations at single-cell resolution during a complex epithelial folding event in the early Drosophila embryo. We expect this tool to be broadly useful for the analysis of epithelial cell geometries and movements in a wide variety of developmental contexts.


Biofilm-growing intestinal anaerobic bacteria.
Sessile growth of anaerobic bacteria from the human intestinal tract has been poorly investigated, so far. We recently reported data on the close association existing between biliary stent clogging and polymicrobial biofilm development in its lumen. By exploiting the explanted stents as a rich source of anaerobic bacterial strains belonging to the genera Bacteroides, Clostridium, Fusobacterium, Finegoldia, Prevotella, and Veillonella, the present study focused on their ability to adhere, to grow in sessile mode and to form in vitro mono- or dual-species biofilms. Experiments on dual-species biofilm formation were planned on the basis of the anaerobic strains isolated from each clogged biliary stent, by selecting those in which a couple of anaerobic strains belonging to different species contributed to the polymicrobial biofilm development. Then, strains were investigated by field emission scanning electron microscopy and confocal laser scanning microscopy to reveal if they are able to grow as mono- and/or dual-species biofilms. As far as we know, this is the first report on the ability to adhere and form mono/dual-species biofilms exhibited by strains belonging to the species Bacteroides oralis, Clostridium difficile, Clostridium baratii, Clostridium fallax, Clostridium bifermentans, Finegoldia magna, and Fusobacterium necrophorum.


Enhanced recovery after surgery for liver transplantation: a review of recent literature.
This review was created to highlight important articles from the past year related to the evolving field of enhanced recovery after surgery (ERAS) in liver transplantation that are relevant to the transplant anesthesiologist. The International Liver Transplantation Society (ILTS) recently completed a landmark project, the ERAS4OLT.org project, which culminated in 80 recommendations for ERAS in liver transplantation. These recommendations encompass management for deceased donor recipients, living donor recipients and living donors. This review highlights selected articles relevant to the perioperative management of deceased donor liver transplant recipients. Though, there are now published internationally agreed upon recommendations for ERAS topics specific to liver transplantation, there is an obvious need for further investigation into this area to provide high quality evidence to support these recommendations. It is reasonable to utilize these recommendations in ERAS protocols for individual institutions; however, more evidence is needed in several areas to confirm the effects of these protocols on short-term outcomes.


The 894G > T (Glu298Asp) variant in the endothelial NOS gene and MTHFR polymorphisms influence homocysteine levels in patients with cognitive decline.
The presence and severity of cerebrovascular pathological findings have been shown to increase the risk and stage of cognitive decline observed in Alzheimer's disease and vascular dementia. Thus, the modification of vascular risk factors seems useful to reduce the risk of dementia regardless of type. Hyperhomocysteinemia has long been known as a major independent risk factor for vascular dysfunction. In this study, we evaluated the relationships between plasma homocysteine levels and genetic risk factors for hyperhomocysteinemia, i.e., the presence of gene variants for methylenetetrahydrofolate reductase (MTHFR) and endothelial nitric oxide synthase (eNOS) in patients with cognitive impairment. Genotyping for MTHFR C677T and eNOS 894G > T polymorphisms was carried out in 69 patients with probable diagnosis of AD and anamnestic mild cognitive impairment, matched for age and gender with 69 healthy volunteers. Patients with MTHFR TT677 genotype showed higher plasma Hcy levels than controls, even after adjustment for folate levels (P < 0.05). Moreover, Hcy plasma levels were higher in cases than controls for any given eNOS genotype. In particular, the presence of eNOS TT894 genotype in patients with cognitive decline resulted significantly associated with increased plasma Hcy levels when compared with controls having the same genotype or patients having other eNOS genotypes (P = 0.02). These data suggest that both MTHFR C677T and eNOS G894T variants should be regarded as genetic risk factors for hyperhomocysteinemia in patients with cognitive decline.


[Ruptured aneurysm of distal posterior inferior cerebellar artery located at caudal loop].
We report an unusual case of subarachnoid hemorrhage (SAH) due to ruptured aneurysm originating at the caudal loop of posterior inferior cerebellar artery (PICA). The patient was a 77-year-old female presented with a sudden onset of headache and vomiting. Initial CT scan demonstrated a SAH with thick hematoma mainly in the cisterna magna. Vertebral angiogram revealed a saccular aneurysm arising from the tonsillomedullary segment of the left PICA, and communicating artery with supplying a territory of contralateral vermis as an anastmotic vessel. At surgery, no vessel branches were confirmed in the vicinity of the aneurysm, and this aneurysm was successfully clipped. In the fetus, numerous basilar and vertebral arteries are organized in plexiform formations around the brain stem. It has been suggested that the pathogenesis of such aneurysm or communicating artery could be related with a remnant of a primitive vertebrobasilar anastomosis. Based on these considerations, congenital vessel-wall weakness and hemodynamic stress associated with communicating artery may contribute to the development of distal PICA aneurysms.


Pilot study of the impact of sporting compression garments on composition and volume of normal and lymphedema legs.
Once clinically manifested as a swollen limb, lymphedema can be difficult to manage. Our focus thus must shift from reactive treatment to proactive management and prevention. On the basis of strong evidence in the literature, lymphedema specialists now encourage exercise as it can improve lymphatic drainage through muscle pump action. However, exercise may increase the lymph load on the vulnerable limb. We aimed to examine whether low level sporting compression is a reasonable recommendation for those with early stage lymphedema by measuring whether sporting compression (SC) tights decrease limb extracellular fluid as measured by Bio-impedance Spectroscopy (BIS) and Perometry in legs following exercise in both healthy controls and those with early stage lymphedema. A group of normal subjects (n=10) and a group of Stage 1 (ISL) lymphedema patients (n=9) were enrolled. Efforts were made to match participants in each group. For those with unilateral lymphedema, the non-affected leg was used as a control. All were measured using BIS, Perometry and Indurometry before and after exercise both with and without sporting compression clothing. The exercise regime was standardized and involved treadmill walking at increasing rates within each person's activity limitation. SC tights were shown to significantly decrease the fluid build up caused by exercise in lymphedema-affected limbs as measured with BIS (p=0.0302). Perometry measurements showed that SC caused a significant decrease in limb volume post exercise of the whole cohort (p=0.0081) and of the control Group B (p=0.0348). Our findings support the notion that SC may provide a socially acceptable and effective means of lymphedema control during exercise for early lymphedema management.


Caries detection accuracy by multiple clinicians and techniques.
This study compared caries detection by multiple clinicians with the accuracy of detection with Caries Detector dye. Four clinicians rated the extent of caries on 100 extracted teeth. After dye application, visual assessment of residual dye was recorded for each specimen. This was repeated after the teeth were embedded in epoxy resin and sectioned. The results indicate that although reasonably accurate overall, clinicians often disagree whether caries is present. The use of a caries detector solution may be a valuable adjunct tool to correct caries diagnosis.


Steady-state H+/O stoichiometry of liver mitochondria.
We have measured the H+/O stoichiometry of rat liver mitochondria respiring in a steady-state, using a novel method. This involves measuring the initial rate of H+ back-flow into mitochondria after respiratory inhibition, with the assumption that this is equal to the steady-state H+-ejection rate. Division by the steady-state O2-consumption rate yields the H+/O ratio. The H+/O values obtained were: 8.3 +/- 1.0 (mean +/- S.E.M.) for 3-hydroxybutyrate: 8.2 +/- 0.7 for glutamate plus malate; 6.0 +/- 0.2 for succinate; 4.1 +/- 0.3 for ascorbate/tetramethylphenylenediamine and 3.0 +/- 0.1 for ascorbate/ferrocyanide. These values correspond to H+/O stoichiometries for electron flow to oxygen from NAD+-linked substrates, succinate and cytochrome c of 8, 6 and 2 (charge/O ratio = 4) respectively.


Risks and benefits of omega 3 fats for mortality, cardiovascular disease, and cancer: systematic review.
To review systematically the evidence for an effect of long chain and shorter chain omega 3 fatty acids on total mortality, cardiovascular events, and cancer. Electronic databases searched to February 2002; authors contacted and bibliographies of randomised controlled trials (RCTs) checked to locate studies. Review of RCTs of omega 3 intake for (3) 6 months in adults (with or without risk factors for cardiovascular disease) with data on a relevant outcome. Cohort studies that estimated omega 3 intake and related this to clinical outcome during at least 6 months were also included. Application of inclusion criteria, data extraction, and quality assessments were performed independently in duplicate. Of 15,159 titles and abstracts assessed, 48 RCTs (36,913 participants) and 41 cohort studies were analysed. The trial results were inconsistent. The pooled estimate showed no strong evidence of reduced risk of total mortality (relative risk 0.87, 95% confidence interval 0.73 to 1.03) or combined cardiovascular events (0.95, 0.82 to 1.12) in participants taking additional omega 3 fats. The few studies at low risk of bias were more consistent, but they showed no effect of omega 3 on total mortality (0.98, 0.70 to 1.36) or cardiovascular events (1.09, 0.87 to 1.37). When data from the subgroup of studies of long chain omega 3 fats were analysed separately, total mortality (0.86, 0.70 to 1.04; 138 events) and cardiovascular events (0.93, 0.79 to 1.11) were not clearly reduced. Neither RCTs nor cohort studies suggested increased risk of cancer with a higher intake of omega 3 (trials: 1.07, 0.88 to 1.30; cohort studies: 1.02, 0.87 to 1.19), but clinically important harm could not be excluded. Long chain and shorter chain omega 3 fats do not have a clear effect on total mortality, combined cardiovascular events, or cancer.


The new Diploma of Rural Generalist Anaesthesia: Supporting Australian rural and remote communities.
In 2023, a Diploma of Rural Generalist Anaesthesia (DipRGA) was implemented across Australia. Developed collaboratively by the Australian and New Zealand College of Anaesthetists (ANZCA), the Australian College of Rural and Remote Medicine (ACRRM) and the Royal Australian College of General Practitioners (RACGP), the 12-month qualification is completed during or following ACRRM or RACGP Rural Generalist Fellowship training. Focused on the needs of rural and remote communities for elective and emergency surgery, maternity care, resuscitative care for medical illness or injury, and stabilisation for retrieval, the DipRGA supports rural generalist anaesthetists working within collaborative teams in geographically isolated settings. The goal is a graduate who can anaesthetise American Society of Anesthesiologists physical status class 1, 2 and stable 3 patients for elective surgery, provide obstetric anaesthesia and analgesia, anaesthetise paediatric patients and undertake advanced crisis care within their scope of practice. Crucially, they also recognise both limitations of their skills and local resources available when considering whether to provide care, defer, refer or transfer patients. DipRGA curriculum design commenced by adapting the ANZCA specialist training curriculum with consideration of the training approach of both the ACRRM and the RACGP, particularly the rural and remote context. Curriculum content is addressed in seven entrustable professional activities supported by workplace-based assessments and multisource feedback. Trainees are supervised by rural generalist anaesthetists and specialist anaesthetists, and complete flexible learning activities to accommodate geographical dispersion. Standardised summative assessments include an early test of knowledge and an examination, adapted from the ACRRM structured assessment using multiple patient scenarios.


Multilevel genomics of colorectal cancers with microsatellite instability-clinical impact of JAK1 mutations and consensus molecular subtype 1.
Approximately 15% of primary colorectal cancers have DNA mismatch repair deficiency, causing a complex genome with thousands of small mutations-the microsatellite instability (MSI) phenotype. We investigated molecular heterogeneity and tumor immunogenicity in relation to clinical endpoints within this distinct subtype of colorectal cancers. A total of 333 primary MSI+ colorectal tumors from multiple cohorts were analyzed by multilevel genomics and computational modeling-including mutation profiling, clonality modeling, and neoantigen prediction in a subset of the tumors, as well as gene expression profiling for consensus molecular subtypes (CMS) and immune cell infiltration. Novel, frequent frameshift mutations in four cancer-critical genes were identified by deep exome sequencing, including in CRTC1, BCL9, JAK1, and PTCH1. JAK1 loss-of-function mutations were validated with an overall frequency of 20% in Norwegian and British patients, and mutated tumors had up-regulation of transcriptional signatures associated with resistance to anti-PD-1 treatment. Clonality analyses revealed a high level of intra-tumor heterogeneity; however, this was not associated with disease progression. Among the MSI+ tumors, the total mutation load correlated with the number of predicted neoantigens (P = 4 × 10-5), but not with immune cell infiltration-this was dependent on the CMS class; MSI+ tumors in CMS1 were highly immunogenic compared to MSI+ tumors in CMS2-4. Both JAK1 mutations and CMS1 were favorable prognostic factors (hazard ratios 0.2 [0.05-0.9] and 0.4 [0.2-0.9], respectively, P = 0.03 and 0.02). Multilevel genomic analyses of MSI+ colorectal cancer revealed molecular heterogeneity with clinical relevance, including tumor immunogenicity and a favorable patient outcome associated with JAK1 mutations and the transcriptomic subgroup CMS1, emphasizing the potential for prognostic stratification of this clinically important subtype. See related research highlight by Samstein and Chan 10.1186/s13073-017-0438-9.


Shape transition of germanium nanocrystals on a silicon (001) surface from pyramids to domes
Chemical vapor deposition of germanium onto the silicon (001) surface at atmospheric pressure and 600 degrees Celsius has previously been shown to produce distinct families of smaller (up to 6 nanometers high) and larger (all approximately 15 nanometers high) nanocrystals. Under ultrahigh-vacuum conditions, physical vapor deposition at approximately the same substrate temperature and growth rate produced a similar bimodal size distribution. In situ scanning tunneling microscopy revealed that the smaller square-based pyramids transform abruptly during growth to significantly larger multifaceted domes, and that few structures with intermediate size and shape remain. Both nanocrystal shapes have size-dependent energy minima that result from the interplay between strain relaxation at the facets and stress concentration at the edges. A thermodynamic model similar to a phase transition accounts for this abrupt morphology change.


Nonviral Expression of LL-37 in a Human Skin Equivalent to Prevent Infection in Skin Wounds.
Inefficient autologous tissue recovery in skin wounds increases the susceptibility of patients to infections caused by multidrug resistant microorganisms, resulting in a high mortality rate. Genetic modification of skin cells has become an important field of study because it could lead to the construction of more functional skin grafts, through the overexpression of antimicrobial peptides that would prevent early contamination and infection with bacteria. In this study, we produce and evaluate human skin equivalents (HSEs) containing transfected human primary fibroblasts and keratinocytes by polyplexes to express the antimicrobial peptide LL-37. The effect of LL-37 on the metabolic activity of normal HSEs was evaluated before the construction of the transfected HSEs, and the antimicrobial efficacy against Pseudomonas aeruginosa and Staphylococcus aureus was evaluated. Subsequently, the levels of LL-37 in the culture supernatants of transfected HSEs, as well as the local expression, were determined. It was found that LL-37 treatment significantly promoted the cellular proliferation of HSEs. Furthermore, HSEs that express elevated levels of LL-37 were shown to possess histological characteristics close to the normal skin and display enhanced antimicrobial activity against S. aureus in vitro. These findings demonstrate that HSEs expressing LL-37 through nonviral modification of skin cells are a promising approach for the prevention of bacterial colonization in wounds.