Identifying important and feasible policies and actions for health at community sports clubs: a consensus-generating approach.
Children's high participation in organised sport in Australia makes sport an ideal setting for health promotion. This study aimed to generate consensus on priority health promotion objectives for community sports clubs, based on informed expert judgements. Delphi survey using three structured questionnaires. Forty-six health promotion, nutrition, physical activity and sport management/delivery professionals were approached to participate in the survey. Questionnaires used an iterative process to determine aspects of sports clubs deemed necessary for developing healthy sporting environments for children. Initially, participants were provided with a list of potential standards for a range of health promotion areas and asked to rate standards based on their importance and feasibility, and any barriers to implementation. Subsequently, participants were provided with information that summarised ratings for each standard to indicate convergence of the group, and asked to review and potentially revise their responses where they diverged. In a third round, participants ranked confirmed standards by priority. 26 professionals completed round 1, 21 completed round 2, and 18 completed round 3. The highest ranked standards related to responsible alcohol practices, availability of healthy food and drinks at sports canteens, smoke-free club facilities, restricting the sale and consumption of alcohol during junior sporting activities, and restricting unhealthy food and beverage company sponsorship. Identifying and prioritising health promotion areas that are relevant to children's sports clubs assists in focusing public health efforts and may guide future engagement of sports clubs. Approaches for providing informational and financial support to clubs to operationalise these standards are proposed.


Fatal ethanol intoxication from household products not intended for ingestion.
Fatal acute ethanol intoxication is frequently encountered in medicolegal practice. Although the vast majority of acute ethanol toxicity deaths follow the ingestion of conventional alcoholic beverages, ethanol can be obtained from a variety of commercial products, which often contain high levels of ethyl alcohol but are not manufactured or designed for consumption. Such products may be easily purchased in locales where statutory limitations restrict liquor availability on Sundays or during the early morning hours. Several acute ethanol fatalities have been encountered in New Mexico that were directly related to consumption of non-beverage ethanol-containing products, all of them occurring during times when alcoholic beverage sales were restricted. Despite the fact that manufacturers deliberately include compounds in these products that discourage ingestion, this policy apparently does little to deter individuals who are searching for a source of ethanol when no conventional beverages are available. The products that were consumed in these fatalities also contained other compounds which would be toxic at much greater concentrations, but which were inconsequential in their effects in comparison with the direct toxic effect of ethanol. Investigation of the scene and awareness that alcohol-containing products can be fatally abused are essential to detecting these unconventional ethanol sources.


Dietary patterns, assessed from a weighed food record, and survival among elderly participants from the United Kingdom.
There is variability in the association between dietary intake and health outcomes across different countries, especially among the elderly. We used the gold standard dietary assessment method, a weighed food record, to examine the association between dietary pattern and mortality in a representative sample of community dwelling participants from Great Britain aged 65 years and older. Dietary intake was recorded at baseline in 1017 elderly participants (520 men, 497 women, mean age 76.3+/-7.4 years). Exploratory factor analysis was performed to examine dietary patterns and participants were followed up over an average of 9.2 years for mortality. The factor analysis revealed four interpretable principal components accounting for approximately 9.8% of the total variance, with similar patterns across sex. A 'Mediterranean-style' dietary pattern explained the greatest proportion of the variance (3.7%), followed by 'health-aware' (2.2%), 'traditional' (2.0%) and 'sweet and fat' (1.9%) factors. There were a total of 683 deaths through follow-up. After adjustment for potential confounders, only the Mediterranean-style dietary pattern remained associated with mortality (highest vs lowest tertile; hazard ratio=0.82, 95% CI, 0.68-1.00). The benefits of the Mediterranean-style diet were only observed among women (hazard ratio=0.71, 95% CI 0.52-0.96) although in men the traditional diet was a risk factor for mortality (hazard ratio=1.30, 95% CI 1.00-1.71). Using a gold standard approach, our results confirm previous evidence that dietary patterns are important in longevity among the elderly.


Avian Reovirus Protein p17 Functions as a Nucleoporin Tpr Suppressor Leading to Activation of p53, p21 and PTEN and Inactivation of PI3K/AKT/mTOR and ERK Signaling Pathways.
Avian reovirus (ARV) protein p17 has been shown to regulate cell cycle and autophagy by activation of p53/PTEN pathway; nevertheless, it is still unclear how p53 and PTEN are activated by p17. Here, we report for the first time that p17 functions as a nucleoporin Tpr suppressor that leads to p53 nuclear accumulation and consequently activates p53, p21, and PTEN. The nuclear localization signal (119IAAKRGRQLD128) of p17 has been identified for Tpr binding. This study has shown that Tpr suppression occurs by p17 interacting with Tpr and by reducing the transcription level of Tpr, which together inhibit Tpr function. In addition to upregulation of PTEN by activation of p53 pathway, this study also suggests that ARV protein p17 acts as a positive regulator of PTEN. ARV p17 stabilizes PTEN by stimulating phosphorylation of cytoplasmic PTEN and by elevating Rak-PTEN association to prevent it from E3 ligase NEDD4-1 targeting. To activate PTEN, p17 is able to promote β-arrestin-mediated PTEN translocation from the cytoplasm to the plasma membrane via a Rock-1-dependent manner. The accumulation of p53 in the nucleus induces the PTEN- and p21-mediated downregulation of cyclin D1 and CDK4. Furthermore, Tpr and CDK4 knockdown increased virus production in contrast to depletion of p53, PTEN, and LC3 reducing virus yield. Taken together, our data suggest that p17-mediated Tpr suppression positively regulates p53, PTEN, and p21 and negatively regulates PI3K/AKT/mTOR and ERK signaling pathways, both of which are beneficial for virus replication.


Rhythmic expression of circadian clock genes in human leukocytes and beard hair follicle cells.
Evaluating individual circadian rhythm traits is crucial for understanding the human biological clock system. The present study reports characterization of physiological and molecular parameters in 13 healthy male subjects under a constant routine condition, where interfering factors were kept to minimum. We measured hormonal secretion levels and examined temporal expression profiles of circadian clock genes in peripheral leukocytes and beard hair follicle cells. All 13 subjects had prominent daily rhythms in melatonin and cortisol secretion. Significant circadian rhythmicity was found for PER1 in 9 subjects, PER2 in 3 subjects, PER3 in all 13 subjects, and BMAL1 in 8 subjects in leukocytes. Additionally, significant circadian rhythmicity was found for PER1 in 5 of 8 subjects tested, PER2 in 2 subjects, PER3 in 6 subjects, and BMAL1 in 3 subjects in beard hair follicle cells. The phase of PER1 and PER3 rhythms in leukocytes correlated significantly with that of physiological rhythms. Our results demonstrate that leukocytes and beard hair follicle cells possess an endogenous circadian clock and suggest that PER1 and PER3 expression would be appropriate biomarkers and hair follicle cells could be a useful tissue source for the evaluation of biological clock traits in individuals.


The real burden of restless legs syndrome: clinical and economic outcomes.
Restless legs syndrome (RLS) is a highly prevalent and substantially underdiagnosed sensorimotor disorder. Only relatively recently have the large impact on patient quality of life (QoL) and the economic burden associated with RLS become more widely recognized. QoL in patients with RLS has been shown to be worse than that of many other chronic conditions, including type 2 diabetes, clinical depression, and osteoarthritis. Sleep disturbance, a cardinal feature of RLS, is the most common and most destructive of its symptoms. More than two-thirds of RLS patients experience serious insomnia, and waking up several times per night is typical for this patient population. Moreover, RLS disrupts rest during waking hours, such as when the patient is sitting or relaxing. Thus, whether awake or asleep, the RLS patient finds little opportunity for the general restorative behaviors necessary for healthy human functioning, resulting in high rates of comorbidities including depression, anxiety, and hypertension. The direct and indirect costs related to RLS have been evaluated in a few studies. Although the cost studies are associated with certain limitations (eg, use of questionnaires), the results show that costs related to RLS are substantial. Healthcare utilization, primarily in the form of doctor visits, constitutes the largest proportion of direct expenditures for RLS in the United States. Indirect costs are also large, primarily due to productivity losses, which are as high as 20% in RLS patients. Effective treatment of RLS is necessary to limit the negative effects of RLS on QoL and to reduce costs associated with the condition.


Two signals are required for negative selection of CD4+CD8+ thymocytes.
Recent results indicate that two signals are required for activation of mature T cells. The first is delivered through the TCR, and the second is delivered through receptors that bind various ligands expressed on APC. For example, it has been shown that B7/BB1, which is expressed on many APC, can costimulate T cell activation by binding to CD28 or CTLA4, which are expressed on mature T cells. In contrast, little is known of the signals required for negative selection of autoreactive thymocytes. Thus, we have investigated this issue by using an in vitro culture system in which thymocytes from mice that are transgenic for a class II MHC-restricted TCR are cultured with murine fibroblast lines that express class II MHC. Under these conditions, CD4+CD8+ (DP) thymocytes undergo an Ag-dependent programmed cell death, which likely represents the negative selection of autoreactive thymocytes that would occur in an intact thymus. Using this culture system, we first found that both TCR- and APC-dependent stimuli were required in order to induce deletion of DP thymocytes. Anti-TCR antibodies alone did not cause deletion of DP cells, but merely induced a decrease in their expression of CD4 and CD8 to produce a DPdull phenotype. Addition of APC was then required for deletion of these DPdull cells. One obvious candidate for the costimulatory signal expressed by these APC was B7. Three different experimental approaches indicated, however, that B7 was not the APC-dependent signal required for deletion of DP thymocytes. Thus, these results suggest that negative selection of autoreactive thymocytes is a two-step process in which stimulation of the TCR causes downregulation of CD4 and CD8 on DP thymocytes, and then an unknown ligand expressed on APC stimulates a receptor on DP thymocytes to induce their deletion.


Plasma angiopoietin-2 predicts the onset of acute lung injury in critically ill patients.
Current clinical prediction scores for acute lung injury (ALI) have limited positive predictive value. No studies have evaluated predictive plasma biomarkers in a broad population of critically ill patients or as an adjunct to clinical prediction scores. To determine whether plasma angiopoietin-2 (Ang-2), von Willebrand factor (vWF), interleukin-8 (IL-8), and/or receptor for advanced glycation end products (sRAGE) predict ALI in critically ill patients. Plasma samples were drawn from critically ill patients (n = 230) identified in the emergency department. Patients who had ALI at baseline or in the subsequent 6 hours were excluded, and the remaining patients were followed for development of ALI. Nineteen patients developed ALI at least 6 hours after the sample draw. Higher levels of Ang-2 and IL-8 were significantly associated with increased development of ALI (P = 0.0008, 0.004, respectively). The association between Ang-2 and subsequent development of ALI was robust to adjustment for sepsis and vasopressor use. Ang-2 and the Lung Injury Prediction Score each independently discriminated well between those who developed ALI and those who did not (area under the receiver operating characteristic curve, 0.74 for each), and using the two together improved the area under the curve to 0.84 (vs. 0.74, P = 0.05). In contrast, plasma levels of sRAGE and vWF were not predictive of ALI. Plasma biomarkers such as Ang-2 can improve clinical prediction scores and identify patients at high risk for ALI. In addition, the early rise of Ang-2 emphasizes the importance of endothelial injury in the early pathogenesis of ALI.


The rate-determining step in cAMP-mediated exocytosis in the rat parotid and submandibular glands appears to involve analogous 26-kDa integral membrane phosphoproteins.
The possible direct involvement of protein phosphorylation in the regulation of exocytosis during beta-adrenergic receptor stimulation in rat parotid and submandibular salivary glands was investigated in vitro using dispersed cells. The dispersed cells were labeled with [32P]orthophosphate for 40 min prior to experimental manipulation. Subcellular fractions were isolated, the proteins were separated using sodium dodecylsulfate/polyacrylamide gel electrophoresis (NaDodSO4/PAGE), and the phosphoproteins were detected by autoradiography. Changes in the extent of phosphorylation for each phosphoprotein were determined indirectly by densitometric analyses. The analogous parotid and submandibular 26-kDa membrane phosphoproteins had a rapid phosphate turnover rate (t 1/2 = 5-6 min) whereas the analogous 21-kDa membrane phosphoproteins had a much slower phosphate turnover rate (t 1/2 greater than 20 min). The results of Triton X-114 extraction indicated that the 26- and 21-kDa phosphoproteins were integral membrane proteins. The rate of phosphate turnover for the analogous 26-kDa phosphoproteins is compatible with a regulatory role in exocytosis, whereas the slower phosphate turnover rate for the analogous 21-kDa phosphoproteins suggests that these proteins may play a more subordinate role in secretion or they may coordinate secretion with other cellular metabolic events.


A Study Protocol for a Parallel-Designed Trial Evaluating the Impact of Plant-Based Diets in Comparison to Animal-Based Diets on Health Status and Prevention of Non-communicable Diseases-The Nutritional Evaluation (NuEva) Study.
Background and Aims: Currently, there is a continuing upward trend for plant-based lifestyles in Germany and Europe. The implementation of vegetarian and vegan lifestyles is characterized by omitting defined food groups such as fish, meat, sausage (vegetarians), or dairy products and honey (vegans). This carries the risk of an undersupply of valuable nutrients. The NuEva study is designed to examine this hypothesis and to evaluate the impact of plant-based diets on health status and disease risk. Methods: The NuEva study is a parallel-designed trial with at least 55 participants for each diet (vegetarian, vegan, flexitarian [rare meat/sausage consumption, once or twice per week]), and participants who consume a traditional Western diet as the control group. In the screening period critical nutrients are identified for the studied diets by analysis of a broad spectrum of nutrients in the human samples (fatty acids, vitamins, minerals, trace elements, nutrient metabolites). Results: Based on the data from the screening period, defined menu plans, ensuring an adequate nutrient intake in accordance with the nutritional guidelines are prepared for each group. The plans are adapted and personalized to individual energy requirements based on the basal metabolic rate and physical activity level. The compliance with the NuEva concept and their impact on nutrient status and cardiovascular risk factors are validated during the intervention period of the NuEva study over 1 year. To investigate the impact of the studied diets on the microbiome, feces samples are collected at the beginning and after the 12 months intervention period (follow up: 12 months). Conclusion: The NuEva study is designed to investigate the impact of common diets on health and disease status, with focus on prevention of cardiovascular diseases. In addition, the effectiveness of the prepared nutritional coaching strategy, ensuring optimal nutrient intake in accordance with the guidelines, is validated during the intervention period of the NuEva study. Clinical Trial Registration: Registered under ClinicalTrials.gov Identifier no. NCT03582020.


Heterotrifunctional chemical cross-linking mass spectrometry confirms physical interaction between human frataxin and ISU.
The progressive neurodegenerative disease Friedreich's ataxia is caused by a decreased level of expression of frataxin, a putative iron chaperone. Frataxin is thought to transiently interact with ISU, the scaffold protein onto which iron-sulfur clusters are assembled, to deliver ferrous iron. Photoreactive heterotrifunctional chemical cross-linking confirmed the interaction between frataxin and ISU in the presence of iron and validated that transient interactions can be covalently trapped with this method. Because frataxin may participate in transient interactions with other mitochondrial proteins, this cross-linking approach may reveal new protein partners and pathways in which it interacts and help deduce direct, downstream consequences of its deficiency.


Validation of a minimum outcome core set in the evaluation of patients with back pain.
Prospective study of patients with subacute osteoporotic fracture (SOF) or chronic low back pain (CLBP). To evaluate reliability, validity, and responsiveness of a purposefully brief outcome instrument. A minimum standardized "core set" was proposed for monitoring patients with low back pain in 1998, but an assessment of metric properties was still lacking. The Core Set, SF-36, and Oswestry questionnaires were completed by 154 patients. Test-retest reproducibility was evaluated in a subsample of 43 stable patients with CLBP. Responsiveness was evaluated by estimating effect size (ES) of pre-postsurgery changes in 50 patients with SOF and 23 with CLBP. The total Core Set showed good reproducibility with intraclass correlation coefficients on test-retest near the highest standard of 0.9, whereas internal consistency differed between patients with CLBP and those with SOF (Cronbach's alpha of 0.92 and 0.64, respectively). Most correlations of the Core Set with SF-36 and Oswestry, previously hypothesized as high, were >0.65, demonstrating good construct validity. Sensitivity to change of the Core Set (ES 0.4-2.3) is similar to the Oswestry (ES 0.7 and 2.3). These findings support the potential usefulness of the Core Set when respondent burden is a major concern. However, subscale scores need to be further tested in other populations before they can be widely recommended.


Overuse tendinosis, not tendinitis part 2: applying the new approach to patellar tendinopathy.
Patellar tendinopathy causes substantial morbidity in both professional and recreational athletes. The condition is most common in athletes of jumping sports such as basketball and volleyball, but it also occurs in soccer, track, and tennis athletes. The disorder arises most often from collagen breakdown rather than inflammation, a tendinosis rather than a tendinitis. Physicians must address the degenerative pathology underlying patellar tendinopathy because regimens that seek to minimize (nonexistent) inflammation would appear illogical. Suggestions for applying the 'tendinosis paradigm' to patellar tendinopathy management include conservative measures such as load reduction, strengthening exercises, and massage. Surgery should be considered only after a long-term and appropriate conservative regimen has failed.


Significantly high levels of ultraviolet-specific mutations in the smoothened gene in basal cell carcinomas from DNA repair-deficient xeroderma pigmentosum patients.
The Sonic hedgehog (SHH) pathway is implicated in the etiology of the most common human cancer in Caucasians, the basal cell carcinoma (BCC). Mutations in the receptor of SHH, the patched gene, have been characterized in sporadic BCCs as well as those from patients with the rare genetic syndromes nevoid BCC and xeroderma pigmentosum (XP). To elucidate the role of UV in the deregulation of the SHH pathway, we analyzed for alterations of smoothened, a transmembrane signaling component regulated by patched, in BCCs and squamous cell carcinomas from UV hypersensitive XP patients. We find UV-specific smoothened mutations in 30% of XP BCCs, three times higher than those in sporadic Caucasian BCCs, confirming the high rate of UV-induced mutations in DNA repair-deficient XP patients. No alteration was found in XP squamous cell carcinomas, indicating the involvement of smoothened specifically in the development of BCC.


Stim2-Eb3 Association and Morphology of Dendritic Spines in Hippocampal Neurons.
Mushroom spines form strong synaptic contacts and are essential for memory storage. We have previously demonstrated that neuronal store-operated calcium entry (nSOC) in hippocampal neurons is regulated by STIM2 protein. This pathway plays a key role in stability of mushroom spines and is compromised in different mice models of Alzheimer's disease (AD). Actin was thought to be the sole cytoskeleton compartment presented in dendritic spines, however, recent studies demonstrated that dynamic microtubules with EB3 capped plus-ends transiently enter spines. We showed that STIM2 forms an endoplasmic reticulum (ER) Ca2+ -dependent complex with EB3 via Ser-x-Ile-Pro aminoacid motif and that disruption of STIM2-EB3 interaction resulted in loss of mushroom spines in hippocampal neurons. Overexpression of EB3 causes increase of mushroom spines fraction and is able to restore their deficiency in hippocampal neurons obtained from PS1-M146V-KI AD mouse model. STIM2 overexpression failed to restore mushroom dendritic spines after EB3 knockdown, while in contrast EB3 overexpression rescued loss of mushroom spines resulting from STIM2 depletion. We propose that EB3 is involved in regulation of dendritic spines morphology, in part due to its association with STIM2, and that modulation of EB3 expression is a potential way to overcome synaptic loss during AD.


CD20 expression in Hodgkin and Reed-Sternberg cells of classical Hodgkin's disease: associations with presenting features and clinical outcome.
CD20 can be expressed in Hodgkin and Reed-Sternberg (HRS) cells of classical Hodgkin's disease (HD), but its clinical significance remains controversial. Therefore, we correlated CD20 expression with presenting features and clinical outcome of untreated patients with classical HD. Patients were eligible if they were previously untreated and human immunodeficiency virus-1 negative, had biopsy-proven classical HD, and if pretreatment paraffin-embedded tumor tissue was available. CD20 expression was determined by immunohistochemistry without knowledge of clinical outcome. A tumor was considered positive if any HRS cells expressed CD20, but other cutoffs for number of CD20-positive HRS were also investigated. We identified 598 patients whose median age was 30 years and of whom 55% were male. HRS cells expressed CD20 in 132 (22%) of 598 patients with classical HD. When any percentage of CD20 expression in HRS cells was used as a cutoff, the 5-year failure-free survival (FFS) for positive versus negative tumors was 86% versus 84%, respectively, for 302 patients treated with doxorubicin, bleomycin, vinblastine, and dacarbazine or equivalent regimens (P =.7 by log-rank test), 74% versus 77%, respectively, for 181 patients treated with mitoxantrone, vincristine, vinblastine, and prednisone and radiotherapy (P =.7 by log-rank test), 74% versus 84%, respectively, for 54 patients treated with MOPP (P =.4 by log-rank test), and 77% versus 88% for 53 patients treated only with radiotherapy (P =.5 by log-rank test). The 5-year FFS was not statistically different when cutoffs of 5% up to 50% for CD20-positive HRS cells were used. CD20 is expressed by HRS cells in 22% of patients with classical HD but is not associated with different FFS after treatment with equivalent regimens.


Intergenerational response to sperm competition risk in an invasive mammal.
Studies of socially mediated phenotypic plasticity have demonstrated adaptive male responses to the 'competitive' environment. Despite this, whether variation in the paternal social environment also influences offspring reproductive potential in an intergenerational context has not yet been examined. Here, we studied the descendants of wild-caught house mice, a destructive pest species worldwide, to address this knowledge gap. We analysed traits that define a 'competitive' phenotype in the sons of males (sires) that had been exposed to either a high-male density (competitive) or high-female density (non-competitive) environment. We report disparate reproductive strategies among the sires: high-male density led to a phenotype geared for competition, while high-female density led to a phenotype that would facilitate elevated mating frequency. Moreover, we found that the competitive responses of sires persisted in the subsequent generation, with the sons of males reared under competition having elevated sperm quality. As all sons were reared under common-garden conditions, variation in their reproductive phenotypes could only have arisen via nongenetic inheritance. We discuss our results in relation to the adaptive advantage of preparing sons for sperm competition and suggest that intergenerational plasticity is a previously unconsidered aspect in invasive mammal fertility control.


Resting Heart Rate Variability (HRV) in Adolescents and Young Adults from a Genetically-Informed Perspective.
Reduced heart rate variability (HRV) is associated with cardiac morbidity, mortality, and negative psychopathology. Most research concerning genetic influences on HRV has focused on adult populations, with fewer studies investigating the developmental period of adolescence and emerging adulthood. The current study estimated the genetic and environmental contributions to resting HRV in a sample of twins using various HRV time domain metrics to assess autonomic function across two different time measurement intervals (2.5- and 10-min). Five metrics of resting HRV [mean interbeat interval (IBI), the standard deviation of normal IBIs (SDNN), root square mean of successive differences between IBIs (RMSSD), cardiac vagal index (CVI), and cardiac sympathetic index (CSI)] were assessed in 421 twin pairs aged 14-20 during a baseline electrocardiogram. This was done for four successive 2.5-min intervals as well as the overall 10-min interval. Heritability (h2) appeared consistent across intervals within each metric with the following estimates (collapsed across time intervals): mean IBI (h2 = 0.36-0.46), SDNN (h2 = 0.23-0.30), RMSSD (h2 = 0.36-0.39), CVI (h2 = 0.37-0.42), CSI (h2 = 0.33-0.46). Beyond additive genetic contributions, unique environment also was an important influence on HRV. Within each metric, a multivariate Cholesky decomposition further revealed evidence of genetic stability across the four successive 2.5-min intervals. The same models showed evidence for both genetic and environmental stability with some environmental attenuation and innovation. All measures of HRV were moderately heritable across time, with further analyses revealing consistent patterns of genetic and environmental influences over time. This study confirms that in an adolescent sample, the time interval used (2.5- vs. 10-min) to measure HRV time domain metrics does not affect the relative proportions of genetic and environmental influences.


Cytochrome oxidase subunit III from Arbacia lixula: detection of functional constraints by comparison with homologous sequences.
In this paper we report the comparison of the sequences of the cytochrome oxidase subunit III from three different sea urchin species. Both nucleotide and amino acid sequences have been analyzed. The nucleotide sequence analysis reveals that the sea urchin sequences obey some rules already found in mammals. The base substitution analysis carried out on the sequences of the three species pairs, shows that the evolutionary dynamics of the first and the second codon positions are so slow that do not allow a quantitative measurement of their genetic distances, thus demonstrating that also in these species the COIII gene is strongly conserved during evolution. Changes occurring at the third codon positions indicate that the three species evolved from a common ancestor under different directional mutational pressure. The multi-alignment of the sea urchin proteins indicates the existence of the amino acid sequence motif N R T that represents a possible glycosylation site. Another glycosylation site has been detected in the mammalian cytochrome oxidase subunit III, in a position slightly different. Such an analysis revealed, for the first time, a new functional aspect of this sequence.


Hapke-based computational method to enable unmixing of hyperspectral data of common salts.
Environmental scientists are currently assessing the ability of hyper-spectral remote sensing to detect, identify, and analyze natural components, including minerals, rocks, vegetation and soil. This paper discusses the use of a nonlinear reflectance model to distinguish multicomponent particulate mixtures. Analysis of the data presented in this paper shows that, although the identity of the components can often be found from diagnostic wavelengths of absorption bands, the quantitative abundance determination requires knowledge of the complex refractive indices and average particle scattering albedo, phase function and size. The present study developed a method for spectrally unmixing halite and gypsum combinations. Using the known refractive indexes of the components, and with the assistance of Hapke theory and Legendre polynomials, the authors develop a method to find the component particle sizes and mixing coefficients for blends of halite and gypsum. Material factors in the method include phase function parameters, bidirectional reflectance, imaginary index, grain sizes, and iterative polynomial fitting. The obtained Hapke parameters from the best-fit approach were comparable to those reported in the literature. After the optical constants (n, the so-called real index of refraction and k, the coefficient of the imaginary index of refraction) are derived, and the geometric parameters are determined, single-scattering albedo (or ω) can be calculated and spectral unmixing becomes possible.


Three classes of motion in the dynamic neutron-scattering susceptibility of a globular protein.
A simplified description of the 295 K dynamics of a globular protein over a wide frequency range (1-1000 GHz) is obtained by combining neutron scattering of lysozyme with molecular dynamics simulation. The molecular dynamics simulation agrees quantitatively with experiment for both the protein and the hydration water and shows that, whereas the hydration water molecules subdiffuse, the protein atoms undergo confined motion decomposable into three distinct classes: localized diffusion, methyl group rotations, and jumps. Each of the three classes gives rise to a characteristic neutron susceptibility signal.


Adenosine triphosphate consumption by bacterial arginyl-transfer ribonucleic acid synthetases.
ATP consumption by arginyl-tRNA synthetases from Escherichia coli and Bacillus stearothermophilus has been investigated by the firefly luciferin--luciferase assay. Arginyl-tRNA synthetase from E. coli utilizes ATP only for aminocylation of tRNA with a 1:1 stoicheiometry. In contrast, we have shown an adenosine triphosphatase activity of arginyl-tRNA synthetase from B. stearothermophilus in the absence of tRNAArg. Dowex chromatography revealed the formation of ADP by the thermophile enzyme; under aminoacylation conditions, AMP was also formed in amounts stoicheiometric with arginyl-tRNA formation.


Interferon regulatory factor-1 up-regulates angiotensin II type 2 receptor and induces apoptosis.
The expression of the angiotensin II type 2 (AT2) receptor is developmentally and growth regulated. In cultured R3T3 cells, expression of this receptor is markedly induced at the confluent state and with serum deprivation. In this study we demonstrated that the removal of serum from culture media resulted in the induction of apoptosis in these cells and the addition of angiotensin II further enhanced apoptosis. We have previously identified an interferon regulatory factor (IRF) binding motif in the mouse AT2 receptor gene promoter region. In this report, we observed that serum removal increased IRF-1 expression, with a rapid and transient decrease of IRF-2. To prove that the changes in IRFs after serum removal mediated apoptosis and up-regulated AT2 receptor, we transfected antisense oligonucleotides for IRF-1 or IRF-2 into R3T3 cells and observed that IRF-1 antisense oligonucleotide attenuated apoptosis and abolished the up-regulation of AT2 receptor. IRF-2 antisense oligonucleotide pretreatment did not affect the onset of apoptosis after serum removal; instead, it increased AT2 receptor binding and enhanced angiotensin II-mediated apoptosis. Taken together, these results suggest that increased IRF-1 after serum starvation contributes to the induction of apoptosis and that increased IRF-1 up-regulates the AT2 receptor expression after serum starvation, resulting in enhanced angiotensin II-mediated apoptosis.


Preschool based JASPER intervention in minimally verbal children with autism: pilot RCT.
In this pilot study, we tested the effects of a novel intervention (JASPER, Joint Attention Symbolic Play Engagement and Regulation) on 3 to 5 year old, minimally verbal children with autism who were attending a non-public preschool. Participants were randomized to a control group (treatment as usual, 30 h of ABA-based therapy per week) or a treatment group (substitution of 30 min of JASPER treatment, twice weekly during their regular program). A baseline of 12 weeks in which no changes were noted in core deficits was followed by 12 weeks of intervention for children randomized to the JASPER treatment. Participants in the treatment group demonstrated greater play diversity on a standardized assessment. Effects also generalized to the classroom, where participants in the treatment group initiated more gestures and spent less time unengaged. These results provide further support that even brief, targeted interventions on joint attention and play can improve core deficits in minimally verbal children with ASD.


Cardiovascular effects of total hip placement in man. With observations on the effects of methylmethacrylate on the isolated rabbit heart.
The cardiovascular effects of total hip placement were evaluated in 10 surgical patients, aged 55 to 82, while receiving fluroxene-N2O-O2 anesthesia. The anesthetic regimen caused mild cardiovascular depression. The placement of the acrylic cement into the acetabulum and femoral shaft also induced mild cardiovascular depression, but these changes were not significant at p less than 0.05. In one 67-yr-old woman, there were significant reductions of cardiac output and stroke volume 2 min after the insertion of acrylic into the femoral shaft, despite careful replacement of intravascular loss and careful anesthetic management. Methylmethacrylate (1 X 10(-6) to 1 X 10(-4), v/v) was administered to 24 isolated perfused rabbits hearts. These concentrations of methylmetacrylate are of the same order as measurable blood levels in surgical patients. There was a dose-dependent depression of left ventricular dP/dt correlated with a depression of the spontaneous heart rate. When the bradycardia was prevented by electrically pacing the hearts or the administration of atropine, the depressed dP/dt rose to control levels. Reduction in myocardial temperature and heart rate by means of reduction in perfusate temperature of the isolated hearts reduced the myocardial depressant effect of methylmethacrylate.


HYPOTHESIS FOR A PRESSURE-SENSITIVE MECHANISM IN MUSCLE SPINDLES.
The suggestion is made that the mammalian muscle spindle may serve in part as a pressure sensor. Pressure acting nonuniformly on the spindle would express the intracapsular fluid distad, thus elongating the capsule. "Intracapsular fibers," those intrafusal muscle fibers inserting into the capsule, would be lengthened, whereas "percapsular fibers" passing beyond the capsule would be relatively unaffected. Since "flower-spray" terminals are preferentially associated with the smaller intrafusal fibers, they would chiefly be concerned with the pressure sensing mechanism. Observations in support of this hypothesis are given.


A statistical pattern recognition approach for determining cellular viability and lineage phenotype in cultured cells and murine bone marrow.
Cellular binding of annexin V and membrane permeability to 7-aminoactinomycin D (7AAD) are important tools for studying apoptosis and cell death by flow cytometry. Combining viability markers with cell surface marker expression is routinely used to study various cell lineages. Current classification methods using strict thresholds, or "gates," on the fluorescent intensity of these markers are subjective in nature and may not fully describe the phenotypes of interest. We have developed objective criteria for phenotypic boundary recognition through the application of statistical pattern recognition. This task was achieved using artificial neural networks (ANNs) that were trained to recognize subsets of cells with known phenotypes, and then used to determine decision boundaries based on statistical measures of similarity. This approach was then used to test the hypothesis that erythropoietin (EPO) inhibits apoptosis and cell death in erythroid precursor cells in murine bone marrow. Our method was developed for classification of viability using an in vitro cell system and then applied to an ex vivo analysis of murine late-stage erythroid progenitors. To induce apoptosis and cell death in vitro, an EPO-dependent human leukemic cell line, UT-7(EPO) cells were incubated without recombinant human erythropoietin (rhEPO) for 72 h. Five different ANNs were trained to recognize live, apoptotic, and dead cells using a "known" subset of the data for training, and a K-fold cross validation procedure for error estimation. The ANNs developed with the in vitro system were then applied to classify cells from an ex vivo study of rhEPO treated mice. Tg197 (human tumor necrosis-alpha transgenic mice, a model of anemia of chronic disease) received a single s.c. dose of 10,000 U/kg rhEPO and femoral bone marrow was collected 1, 2, 4, and 8 days after dosing. Femoral bone marrow cells were stained with TER-119 PE, CD71 APC enable identification of erythroid precursors, and annexin V FITC and 7AAD to identify the apoptotic and dead cells. During classification forward and side angle light scatter were also input to all pattern recognition systems. Similar decision boundaries between live, apoptotic, and dead cells were consistently identified by the neural networks. The best performing network was a radial basis function multi-perceptron that produced an estimated average error rate of 4.5% +/- 0.9%. Using these boundaries, the following results were reached: depriving UT-7(EPO) cells of rhEPO induced apoptosis and cell death while the addition of rhEPO rescued the cells in a dose-dependent manner. In vivo, treatment with rhEPO resulted in an increase of live erythroid cells in the bone marrow to 119.8% +/- 9.8% of control at the 8 day time point. However, a statistically significant transient increase in TER-119(+) CD71(+) 7AAD(+) dead erythroid precursors was observed at the 1 and 2 day time points with a corresponding decrease in TER-119(+) CD71(+) 7AAD(-) Annexin V(-) live erythroid precursors, and no change in the number of TER-119(+) CD71(+) annexin V(+) 7AAD(-) apoptotic erythroid precursors in the bone marrow. A statistical pattern recognition approach to viability classification provides an objective rationale for setting decision boundaries between "positive" and "negative" intensity measures in cytometric data. Using this approach we have confirmed that rhEPO inhibits apoptosis and cell death in an EPO dependent cell line in vitro, but failed to do so in vivo, suggesting EPO may not act as a simple antiapoptotic agent in the bone marrow. Rather, homeostatic mechanisms may regulate the pharmacodynamic response to rhEPO.


High aerodynamic lift from the tail reduces drag in gliding raptors.
Many functions have been postulated for the aerodynamic role of the avian tail during steady-state flight. By analogy with conventional aircraft, the tail might provide passive pitch stability if it produced very low or negative lift. Alternatively, aeronautical principles might suggest strategies that allow the tail to reduce inviscid, induced drag: if the wings and tail act in different horizontal planes, they might benefit from biplane-like aerodynamics; if they act in the same plane, lift from the tail might compensate for lift lost over the fuselage (body), reducing induced drag with a more even downwash profile. However, textbook aeronautical principles should be applied with caution because birds have highly capable sensing and active control, presumably reducing the demand for passive aerodynamic stability, and, because of their small size and low flight speeds, operate at Reynolds numbers two orders of magnitude below those of light aircraft. Here, by tracking up to 20,000, 0.3 mm neutrally buoyant soap bubbles behind a gliding barn owl, tawny owl and goshawk, we found that downwash velocity due to the body/tail consistently exceeds that due to the wings. The downwash measured behind the centreline is quantitatively consistent with an alternative hypothesis: that of constant lift production per planform area, a requirement for minimizing viscous, profile drag. Gliding raptors use lift distributions that compromise both inviscid induced drag minimization and static pitch stability, instead adopting a strategy that reduces the viscous drag, which is of proportionately greater importance to lower Reynolds number fliers.


Factors Associated With Attendance for Cardiac Neurodevelopmental Evaluation.
Neurodevelopmental evaluation of toddlers with complex congenital heart disease is recommended but reported frequency is low. Data on barriers to attending neurodevelopmental follow-up are limited. This study aims to estimate the attendance rate for a toddler neurodevelopmental evaluation in a contemporary multicenter cohort and to assess patient and center level factors associated with attending this evaluation. This is a retrospective cohort study of children born between September 2017 and September 2018 who underwent cardiopulmonary bypass in their first year of life at a center contributing data to the Cardiac Neurodevelopmental Outcome Collaborative and Pediatric Cardiac Critical Care Consortium clinical registries. The primary outcome was attendance for a neurodevelopmental evaluation between 11 and 30 months of age. Sociodemographic and medical characteristics and center factors specific to neurodevelopmental program design were considered as predictors for attendance. Among 2385 patients eligible from 16 cardiac centers, the attendance rate was 29.0% (692 of 2385), with a range of 7.8% to 54.3% across individual centers. In multivariable logistic regression models, hospital-initiated (versus family-initiated) scheduling for neurodevelopmental evaluation had the largest odds ratio in predicting attendance (odds ratio = 4.24, 95% confidence interval, 2.74-6.55). Other predictors of attendance included antenatal diagnosis, absence of Trisomy 21, higher Society of Thoracic Surgeons-European Association for Cardio-Thoracic Surgery mortality category, longer postoperative length of stay, private insurance, and residing a shorter distance from the hospital. Attendance rates reflect some improvement but remain low. Changes to program infrastructure and design and minimizing barriers affecting access to care are essential components for improving neurodevelopmental care and outcomes for children with congenital heart disease.


Housing, HIV outcomes, and related comorbidities in persons living with HIV in Washington, DC.
Housing instability can hinder the ability of people with HIV (PWH) to maintain engagement in care, adhere to antiretroviral (ART) regimens, and achieve viral suppression. This analysis examined the association between housing instability and HIV outcomes, as well as the association between housing status and substance use disorders, mental health disorders, and sexually transmitted infections. Multivariable logistic regressions were performed using data from the DC Cohort, a longitudinal cohort of PWH. Among 8622 PWH, unstably housed PWH were significantly more likely to be prescribed ART (aOR: 1.4; 95%CI: 1.1, 1.8) yet were significantly less likely to be virally suppressed (aOR: 0.6; 95%CI: 0.5, 0.8). Unstably housed PWH were also significantly more likely to have a substance use or mental health disorder, which may inhibit PWH's ability to achieve viral suppression. Efforts to end the HIV epidemic should address housing to ensure treatment is optimized for key populations.


Sex differences in the effect of acute fasting on excitatory and inhibitory synapses onto ventral tegmental area dopamine neurons.
Fasting can increase motivation for food and can energize reward-seeking. Ventral tegmental area (VTA) dopamine neurons respond to motivationally relevant information and fasting can influence mesolimbic dopamine concentration. An acute overnight fast differentially alters food approach behaviours and excitatory synaptic transmission onto VTA dopamine neurons of male or female mice. While inhibitory synapses onto VTA dopamine neurons are not altered by fasting in male or female mice, male mice had strengthened excitatory synapses whereas female mice had increased endocannabinoid-mediated short-term plasticity at excitatory synapses. These results help us understand how fasting differentially influences excitatory synaptic transmission onto dopamine neurons and may inform different strategies for fasting-induced food seeking by male and female mice. Dopamine neurons in the ventral tegmental area (VTA) are important for energizing goal-directed behaviour towards food and are sensitive to changes in metabolic states. Fasting increases the incentive motivation for food and the mobilization of energy stores and has sex-dependent effects. However, it is unknown how acute fasting alters excitatory or inhibitory synaptic transmission onto VTA dopamine neurons. An acute 16 h overnight fast induced increased food-seeking behaviour that was more predominant in male mice. Fasting increased miniature excitatory postsynaptic current frequency and amplitude in male, but not female, mice. This effect was not due to altered release probability as there was no change in the paired pulse ratio, nor was it due to an altered postsynaptic response as there was no change in the AMPA receptor/NMDA receptor ratio or response to glutamate uncaging. However, this effect was consistent with an increase in the number of release sites. In addition, depolarization-induced suppression of excitation, a measure of short-term endocannabinoid-mediated plasticity, was enhanced in female but not male fasted mice. There were no fasting-induced changes at inhibitory synapses onto dopamine neurons of male or female mice. Taken together, these results demonstrate that fasting influences excitatory synapses differentially in male and female mice, but preserves inhibitory synapses onto dopamine neurons, indicating that the mesolimbic circuits of male and female mice respond differently to acute energy deprivation.


Positive projective identification.
This paper suggests that literature concerning projective identification has excessively focused on the projection of unwanted aspects of the self without giving adequate consideration to projective identification involving good qualities. It is proposed that clinical advantages may derive from dividing projective identification into positive and negative components. This division allows a more ready emphasis on the capacity for beneficial object relations which even psychotic individuals retain despite their obvious hostility. Case vignettes are provided to demonstrate positive projective identification and to emphasize the clinical usefulness of this concept.


Iso-osmolar radio contrast iodixanol in patients with chronic kidney disease.
Although radio contrast volume has been associated with worsening post-procedural kidney function, this relationship has not been extensively studied using an iso-osmolar contrast agent in chronic kidney disease patients. We retrospectively studied patients undergoing cardiac catheterization at the University of Minnesota from 2000 to 2004, using the iso-osmolar contrast agent, iodixanol. All patients were included who had calculated creatinine clearance (CCR) < 60 mL/min, not on dialysis, and serum creatinine measured on the same day and within 7 days after the procedure. Comparison of a subgroup with severe chronic kidney disease and diabetes mellitus was compared to a similar historical control group that used the low-osmolar contrast agent, iohexol. Serum creatinine and CCR were 2.9 +/- 1.5 mg/dL and 33.4 +/- 12.0 mL/min (mean +/- standard deviation), respectively, at baseline in 117 cases. Peak creatinine increased by 0.03 +/- 0.7 mg/dL after 84.3 +/- 67.3 mL of iodixanol was used. Contrast-induced nephropathy definition was fulfilled in 22 (18.8%) cases. A non-significant negative correlation was found between the volume of iodixanol and the change in creatinine (r2 = 0.0011, p = 0.7254). A subgroup with severe chronic kidney disease and diabetes mellitus with iodixanol had a significantly lower creatinine increase (n = 25, 0.09 +/- 0.5 mg/dL), compared to historical controls (n = 42, 0.7 +/- 0.8 mg/dL) with iohexol (p < 0.001). A non-significant positive correlation between volume of contrast and change in creatinine was found in this subgroup who received iodixanol (n = 25, r2 = 0.0756, p = 0.1835), but was significant in the historical controls who received iohexol (n = 42, r2 = 0.135, p = 0.017). The volume of iso-osmolar radio contrast does not affect the incidence of contrast-induced nephropathy in patients with chronic kidney disease. A randomized trial evaluating the incidence of contrast nephropathy would verify the safety of ad hoc versus staged angiographic procedures in this population.


Polymer Modeling Reveals Interplay between Physical Properties of Chromosomal DNA and the Size and Distribution of Condensin-Based Chromatin Loops.
Transient DNA loops occur throughout the genome due to thermal fluctuations of DNA and the function of SMC complex proteins such as condensin and cohesin. Transient crosslinking within and between chromosomes and loop extrusion by SMCs have profound effects on high-order chromatin organization and exhibit specificity in cell type, cell cycle stage, and cellular environment. SMC complexes anchor one end to DNA with the other extending some distance and retracting to form a loop. How cells regulate loop sizes and how loops distribute along chromatin are emerging questions. To understand loop size regulation, we employed bead-spring polymer chain models of chromatin and the activity of an SMC complex on chromatin. Our study shows that (1) the stiffness of the chromatin polymer chain, (2) the tensile stiffness of chromatin crosslinking complexes such as condensin, and (3) the strength of the internal or external tethering of chromatin chains cooperatively dictate the loop size distribution and compaction volume of induced chromatin domains. When strong DNA tethers are invoked, loop size distributions are tuned by condensin stiffness. When DNA tethers are released, loop size distributions are tuned by chromatin stiffness. In this three-way interaction, the presence and strength of tethering unexpectedly dictates chromatin conformation within a topological domain.


Beating Heart Validation of Safety and Efficacy of a Percutaneous Pericardiotomy Tool.
Epicardial procedures frequently require pericardial manipulation. We aimed to develop a nonsurgical percutaneous pericardial modification tool that may (1) facilitate epicardial-based procedures by enabling adhesiolysis or (2) attenuate the myocardial constraining effect of the pericardium. Three novel devices were developed to enable pericardiotomy, all of which can be deployed in over-the-wire fashion following percutaneous epicardial access. The grasper permits us to seize the pericardial membrane providing leverage for incision. The scissors enables anterograde cutting maneuvers. The reverse-slitter allows retrograde incisions; in addition, this device has a deflectable tip that increases the potential cutting area. We optimized these tools for safety by including electrodes to test for phrenic nerve stimulation as well as myocardial stimulation to determine directionality of the cutting devices. The base of the scissors and reverse-slitter are also blunt ensuring that the cutting element is always away from the myocardium. Following 5 nonbeating heart bench test experiments for prototype development, 11 animal (9 canine, 2 swine) studies were performed. Of these 2 were proof-of-concept open chest studies; the remaining 9 were entirely closed-chest, percutaneous procedures allowing for remodification of the prototypes. The tools successfully permitted incision of the pericardium in all studies. Hemodynamic measurements were assessed postincision and showed no compromise of systolic function. No coronary artery or phrenic nerve damage was seen in any study. Percutaneous pericardiotomy is feasible and appears to be safe. It may provide leverage in epicardial-based procedures and offer treatment options in disease processes characterized by pericardial restraint.


Photodissociation and Infrared Spectroscopy of Uranium-Nitrogen Cation Complexes.
Laser vaporization of uranium in a pulsed supersonic expansion of nitrogen is used to produce complexes of the form U+(N2)n (n = 1-8). These ions are mass selected in a reflectron time-of-flight spectrometer and studied with visible and UV laser fixed-frequency photodissociation and with tunable infrared laser photodissociation spectroscopy. The dissociation patterns and spectroscopy of U+(N2)n indicate that N2 ligands are intact molecules and that there is no insertion chemistry resulting in UN+ or NUN+. Fixed frequency photodissociation at 532 and 355 nm indicate that the U+-N2 bond dissociation energy varies little with changing coordination. The photon energy and the number of ligands eliminated allow an estimate of the average U+-N2 dissociation energy of 12 kcal/mol. Infrared bands are observed for these complexes near the N-N stretch vibration via elimination of N2 molecules. These resonances are observed to be shifted about 130 cm-1 to the red from the free-N2 frequency for complexes with n = 3-8. Density functional theory indicates that U+ is most stable in the sextet state in these complexes and that N2 molecules bind in end-on configurations. The fully coordinated complex is predicted to be U+(N2)8, which has a cubic structure. The vibrational frequencies predicted by theory are consistently lower than those in the experiment, independent of the isomeric structure or spin state of the complexes. Despite its failure to reproduce the infrared spectra, theory provides an average U+-N2 dissociation energy of 11.8 ± 0.5 kcal/mol, in good agreement with the value from the experiments.


Comparative effects of SCH 1000 and fenoterol after histamine-induced bronchoconstriction in asymptomatic asthmatics.
In 16 asymptomatic asthmatics a functional study was carried out under controlled conditions during histamine bronchial provocation test (HBPT) and after bronchodilation test with either SCH 1000 or fenoterol by evaluating both SRAW and the slope of the alveolar plateau of nitrogen washout curve (N2 ph III). During HBPT, 9 of 16 subjects showed a central response (increase in RAW and SRAW only), whereas in the remaining 7 subjects histamine induced a mixed response due to an increase in both SRAW and N2 ph III. No differences between the two groups were observed in age and functional data during controlled conditions. Doses of delivered histamine were not significantly different. In 5 of 9 central responders to HBPT, SCH 1000 induced a decrease in SRAW only, whereas in 3 of the remaining 4 fenoterol caused both central and peripheral bronchodilation (decrease in both SRAW and N2 ph III). All mixed responders to HBPT showed a mixed bronchodilation during SCH 1000 (three subjects) or fenoterol (the remaining four). These data seem to indicate that the mechanism of bronchodilation induced by SCH 1000 is related to that of histamine-induced bronchoconstriction. Moreover, bronchodilation due to fenoterol could be related to a vagal modulation or be a systemic direct effect.


Hospital Emergency Readiness Overview study.
In 2001, a survey of Canadian emergency departments indicated significant deficiencies in disaster preparedness. Since then, there have been efforts on the part of Provincial governments to remedy this situation. This survey repeats the original study with minor modifications to determine if there has been improvement. The Hospital Emergency Readiness Overview study demonstrates that despite improvements, there remain gaps in Canadian healthcare facility readiness for disaster, specifically one involving contaminated patients. It also highlights the lack of any standardized assessment of healthcare facilities' chemical, biological, radiological, or nuclear readiness.


[Impaired bronchial patency in patients with idiopathic interstitial pneumonias].
Idiopathic interstitial pneumonias are well-known to be mainly characterized by restrictive ventilation dysfunctions and a diminished lung diffuse capacity. No separate significance is generally attached to bronchial obstructive disorders that may be detected along with restrictive ones in some patients. The made analysis of the ventilation capacity of the lung in 53 patients with idiopathic interstitial pneumonias has revealed impaired bronchial patency in 47.2% of the cases, which gives grounds to study this problem in future.


T1 and T2 measurements of meningiomas and neuromas before and after Gd-DTPA.
Seven patients with meningiomas and five patients with neuromas were examined with spin-echo sequences on a 0.35-T imaging system. Signal enhancement and relaxation rate increments with gadolinium-diethylenetriamine pentaacetic acid (Gd-DTPA) were evaluated, as well as relaxation rate contributions, indicators of Gd-DTPA accessibility to the tissue water. The average signal enhancement rate with Gd-DTPA was higher in neuromas than in meningiomas, 148% and 84%, respectively, but this difference was poorly appreciated on enhanced images because of similar average postcontrast T1 values in both tumors. However, the average T1 relaxation increment was almost twofold higher in neuromas than in meningiomas, 318% and 162%, respectively, mainly deriving from longer intrinsic T1 values in neuromas. Also, there was a 25% increase in the average T2 relaxation rate in neuromas after Gd-DTPA together with a higher T2 contribution by Gd-DTPA, while such effects were hardly discernible in meningiomas, suggestive of greater accessibility of Gd-DTPA to the tissue water in neuromas. By electron microscopy, endothelial fenestration and open gap junctions are commonly found in capillaries of both tumors, functioning as the routes into the extracellular space where the major contrast effect of Gd-DTPA can be expected. However, the open gap junctions are short and straight in neuromas, while they are tortuous and sinusoid in meningiomas. This may provide greater access for contrast material into relatively larger extracellular spaces in neuromas and may cause a greater T1 relaxation rate increment with Gd-DTPA.(ABSTRACT TRUNCATED AT 250 WORDS)


T cells and macrophages in Trypanosoma brucei-related glomerulopathy.
In a previous study, susceptibility for Trypanosoma brucei-related glomerulopathy in mice was shown to be dependent on non-major histocompatibility complex genes. Glomerular disease in this model could not be explained by the production of autoantibodies alone. In order to analyze which part of the defense system, in addition to the B-cell compartment, is involved in the development of this infection-related glomerular disease, groups of athymic (BALB/c rnu/rnu), splenectomized, or macrophage-depleted BALB/c mice were inoculated with T. brucei parasites. Polyclonal B-cell activation, invariably observed in infected BALB/c mice, was absent in BALB/c rnu/rnu mice. Glomerular disease in athymic mice, however, as defined by albuminuria and deposition of immune complexes, was not different from that seen in euthymic infected BALB/c mice. Splenectomy prior to inoculation of parasites led to a decreased incidence of albuminuria in 40% of the animals, whereas splenectomy 21 days after inoculation reduced albuminuria significantly, suggesting a role for spleen cells in the induction of glomerular disease. After macrophage depletion with liposome-encapsulated dichlorodimethylene-diphosphonate, infected BALB/c mice developed significantly higher albuminuria levels for a period up to 2 weeks after depletion. Therefore, it was concluded that the development of T. brucei-related glomerular disease is independent of thymus-matured T cells, while the involvement of macrophages in the development of proteinuria is inhibitory rather than disease inducing. Spleen cells other than thymus-dependent T cells, B cells, and macrophages should be investigated for their role in the pathogenesis of this glomerulopathy.


Time doesn't change everything: the longitudinal course of distress tolerance and its relationship with externalizing and internalizing symptoms during early adolescence.
Although distress tolerance is an emerging construct of empirical interest, we know little about its temporal change, developmental trajectory, and prospective relationships with maladaptive behaviors. The current study examined the developmental trajectory (mean- and individual-level change, and rank-order stability) of distress tolerance in an adolescent sample of boys and girls (N = 277) followed over a four-year period. Next we examined if distress tolerance influenced change in Externalizing (EXT) and Internalizing (INT) symptoms, and if EXT and INT symptoms in turn influenced change in distress tolerance. Finally, we examined if any of these trends differed by gender. Results indicated that distress tolerance is temporally stable, with little mean- or individual-level change. Latent growth models reported that level of distress tolerance is cross-sectionally associated with both EXT and INT symptoms, yet longitudinally, only associated with EXT symptoms. These results suggest that distress tolerance should be a focus of research on etiology and intervention.


Renal biopsy morphology in renal transplantation. A comparative study of the light-microscopic appearances of biopsies from patients treated with cyclosporin A or azathioprine prednisone and antilymphocyte globulin.
Nephrotoxicity is a major side effect of cyclosporin A (CSA) when used in renal transplantation, and the distinction between nephrotoxicity and rejection is important in patient management. One hundred twenty-five renal biopsies were examined from 56 patients entered into a controlled clinical trial aimed at comparing the efficacy of CSA therapy alone to a combination of prednisone, azathioprine, and antilymphocyte globulin (AZA). In order to define the histopathology of rejection and nephrotoxicity, all the biopsies were evaluated in a semiquantitative manner by an observer unaware of the clinical state of the patient. Comparison of the morphological appearances of 32 biopsies from patients on CSA, and 22 biopsies from AZA-treated patients performed during clinically apparent rejection episodes showed that the histological patterns of rejection were the same in both treatment groups. Comparison of the morphological features of 34 biopsies from patients receiving CSA and 13 from patients receiving AZA, performed during prolonged periods of post-transplant renal failure, who eventually recovered on continuation of original therapy, showed that there were no morphological features specific to the CSA-treated group. Five patients on CSA had oliguria which was prolonged by CSA nephrotoxicity. Thirteen biopsies from all five patients showed a diffuse interstitial fibrosis that was peculiar to this group of patients.


Potentiation of nerve growth factor-induced neurite outgrowth in PC12 cells by ifenprodil: the role of sigma-1 and IP3 receptors.
In addition to both the α1 adrenergic receptor and N-methyl-D-aspartate (NMDA) receptor antagonists, ifenprodil binds to the sigma receptor subtypes 1 and 2. In this study, we examined the effects of ifenprodil on nerve growth factor (NGF)-induced neurite outgrowth in PC12 cells. Ifenprodil significantly potentiated NGF-induced neurite outgrowth, in a concentration-dependent manner. In contrast, the α1 adrenergic receptor antagonist, prazosin and the NMDA receptor NR2B antagonist, Ro 25-6981 did not alter NGF-induced neurite outgrowth. Potentiation of NGF-induced neurite outgrowth mediated by ifenprodil was significantly antagonized by co-administration of the selective sigma-1 receptor antagonist, NE-100, but not the sigma-2 receptor antagonist, SM-21. Similarly, ifenprodil enhanced NGF-induced neurite outgrowth was again significantly reduced by the inositol 1,4,5-triphosphate (IP(3)) receptor antagonists, xestospongin C and 2-aminoethoxydiphenyl borate (2-APB) treatment. Furthermore, BAPTA-AM, a chelator of intracellular Ca(2+), blocked the effects of ifenprodil on NGF-induced neurite outgrowth, indicating the role of intracellular Ca(2+) in the neurite outgrowth. These findings suggest that activation at sigma-1 receptors and subsequent interaction with IP(3) receptors may mediate the pharmacological effects of ifenprodil on neurite outgrowth.


Epidermal growth factor receptor tyrosine kinase inhibitors and bone metastases: different mechanisms of action for a novel therapeutic application?
The paper by Angelucci et al. published in the current issue of Endocrine-Related Cancer suggests a potential, novel application of epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) in the treatment of bone metastases. Interestingly, activity of anti-EGFR agents on the pathogenesis and progression of bone metastases has been described in previous reports, and a number of different mechanisms seem to be involved in this phenomenon. Anti-EGFR agents have a direct activity on tumour cells in which they produce growth inhibition, apoptosis, and reduced invasive capacity through the inhibition of molecules associated with tissue invasion such as urokinase-type plasminogen activator (uPA) and matrix metalloproteinase (MMP)-9. In addition, these compounds have an anti-angiogenic activity, either direct by affecting the proliferation and survival of endothelial cells, or indirect by blocking the production of vascular endothelial growth factor (VEGF) in bone marrow stromal cells and in tumour cells. Finally, EGFR-TKIs can inhibit recruitment of osteoclasts in bone lesions, by affecting the ability of bone marrow stromal cells to induce osteoclast differentiation and activation. Taken together, these findings strongly support prospective clinical trials of anti-EGFR agents in cancer patients with bone metastases in order to define their role in the management of bone disease.


Dislocated lens nuclei simulating choroidal melanomas.
Three patients underwent echographic examinations for lesions suspected of being choroidal malignant melanomas. All patients were pseudophakic, and two had extensive optic nerve damage from inflammatory glaucoma. In all three patients echography demonstrated an intravitreal lens nucleus that had been lost during cataract extraction. To our knowledge, dislocated lens nuclei have not been reported previously in the differential diagnosis of choroidal melanoma. This diagnosis should be considered in the aphakic or pseudophakic patient with a fundus mass lesion. Echography is especially helpful in establishing the correct diagnosis.


Performance characteristics of a novel clustered multi-pinhole technology for simultaneous high-resolution SPECT/PET.
Versatile emission computed tomography (VECTor) for small-animal imaging enables fully simultaneous SPECT/PET image acquisition based on clustered multi-pinhole collimation. The present study experimentally evaluated the detailed performance characteristics of the clustered multi-pinhole system for simultaneous (99m)Tc and (18)F imaging from the user perspective. Spatial resolution, sensitivity, count rate linearity were determined for the VECTor system (MILabs). Two hot-rod micro-resolution phantoms with 6 sectors were created to test the resolution of (99m)Tc and (18)F. Sensitivity and count rate linearity were measured by scanning (99m)Tc and (18)F point sources positioned at the center of the field of view. Furthermore, we quantified the influence of (18)F on (99m)Tc SPECT images. The ratios of SPECT counts on the (99m)Tc-only and simultaneous (18)F-(99m)Tc at various time points were evaluated as a function of the (18)F-to-(99m)Tc activity concentration ratio. The 0.5-mm hot-rods can be visually distinguished in the (99m)Tc image, and 0.8-mm rods for (18)F remained clearly visible. The point-source sensitivity was 2800 cps/MBq for (99m)Tc and 2899 cps/MBq for (18)F, respectively. Count rates up to 120,000 cps for one bed position were linear for the activity. Spill-over from (18)F into (99m)Tc SPECT images was negligible when the activity concentration of the administered (18)F solution exceeded that of the (99m)Tc solution by up to a factor of 2. We evaluated the performance characteristics of the VECTor that lead to determination of the optimal administered doses of (99m)Tc and (18)F tracers. We found that the VECTor achieved high resolution and high sensitivity as well as good (99m)Tc and (18)F linearity. Simultaneous SPECT/PET imaging with (99m)Tc and (18)F tracers, and stand-alone (99m)Tc and (18)F imaging using clustered-pinhole collimators is feasible and practical for a wide range of research applications using small animals.


Quantification of pre-mRNA escape rate and synergy in splicing.
Splicing reactions generally combine high speed with accuracy. However, some of the pre-mRNAs escape the nucleus with a retained intron. Intron retention can control gene expression and increase proteome diversity. We calculated the escape rate for the yeast PTC7 intron and pre-mRNA. This prediction was facilitated by the observation that splicing is a linear process and by deriving simple algebraic expressions from a model of co- and post-transcriptional splicing and RNA surveillance that determines the rate of the nonsense-mediated decay (NMD) of the pre-mRNAs with the retained intron. The escape rate was consistent with the observed threshold of splicing rate below which the mature mRNA level declined. When an mRNA contains multiple introns, the outcome of splicing becomes more difficult to predict since not only the escape rate of the pre-mRNA has to be considered, but also the possibility that the splicing of each intron is influenced by the others. We showed that the two adjacent introns in the SUS1 mRNA are spliced cooperatively, but this does not counteract the escape of the partially spliced mRNA. These findings will help to infer promoter activity and to predict the behavior of and to control splicing regulatory networks.


Status Quo and Trends of Intra-Arterial Therapy for Brain Tumors: A Bibliometric and Clinical Trials Analysis.
Intra-arterial drug delivery circumvents the first-pass effect and is believed to increase both efficacy and tolerability of primary and metastatic brain tumor therapy. The aim of this update is to report on pertinent articles and clinical trials to better understand the research landscape to date and future directions. Elsevier's Scopus and ClinicalTrials.gov databases were reviewed in August 2021 for all possible articles and clinical trials of intra-arterial drug injection as a treatment strategy for brain tumors. Entries were screened against predefined selection criteria and various parameters were summarized. Twenty clinical trials and 271 articles satisfied all inclusion criteria. In terms of articles, 201 (74%) were primarily clinical and 70 (26%) were basic science, published in a total of 120 different journals. Median values were: publication year, 1986 (range, 1962-2021); citation count, 15 (range, 0-607); number of authors, 5 (range, 1-18). Pertaining to clinical trials, 9 (45%) were phase 1 trials, with median expected start and completion years in 2011 (range, 1998-2019) and 2022 (range, 2008-2025), respectively. Only one (5%) trial has reported results to date. Glioma was the most common tumor indication reported in both articles (68%) and trials (75%). There were 215 (79%) articles investigating chemotherapy, while 13 (65%) trials evaluated targeted therapy. Transient blood-brain barrier disruption was the commonest strategy for articles (27%) and trials (60%) to optimize intra-arterial therapy. Articles and trials predominately originated in the United States (50% and 90%, respectively). In this bibliometric and clinical trials analysis, we discuss the current state and trends of intra-arterial therapy for brain tumors. Most articles were clinical, and traditional anti-cancer agents and drug delivery strategies were commonly studied. This was reflected in clinical trials, of which only a single study had reported outcomes. We anticipate future efforts to involve novel therapeutic and procedural strategies based on recent advances in the field.


Comparison analysis of optical frequency comb generation with nonlinear effects in highly nonlinear fibers.
We compare several schemes for broadband optical frequency comb (OFC) generation based on several nonlinear effects in highly nonlinear fiber (HNLF). Cascaded four wave mixing (CFWM) and self-phase modulation (SPM) processes in HNLF are proved to be effective ways for spectrum broadening. We investigate some parameters affecting the performance of the output OFC in detail. When only CFWM occurs in the HNLF, broadband OFC can be generated with poor power flatness. When only SPM occurs in the HNLF, we obtain a 10 GHz OFC of 103 comb lines within 5-dB power deviation. When both CFWM and SPM simultaneously occur in the HNLF, we obtain a 10 GHz OFC of 143 comb lines within 4.5-dB power deviation. All the OFC generation schemes have the advantages of tunability of central wavelength and repetition frequency.


Politicizing NIH funding: a bridge to nowhere.
We live in a time of increased spending, mounting debt, and few remedies for balancing the federal budget that have bipartisan support. Unfortunately, one recent target for decreased allocations of the federal budget is the NIH; the discussion of the awarded grants and the grant funding process has been skewed and altered to present medical research in an unfriendly light, and this can have very damaging repercussions. Politicizing this process could ultimately challenge human health, technology, and economic growth.


Thyroid abnormalities following neck irradiation for Hodgkin's disease.
Thyroid function and scans were evaluated in fifty disease-free patients 2 to 16 years after receiving neck irradiation for the treatment of Hodgkin's disease. Twenty-five of 50 patients had abnormal thyroid studies: eight were hypothyroid, two were hypothyroid and had abnormal scans, and fifteen had abnormal scans. Of the 15 patients with abnormal scans, one had an isolated elevation of TSH (thyroid stimulating hormone) and one developed exophthalmos. These data, obtained within a relatively short follow-up period, indicate that morphologic and functional abnormalities of the thyroid gland are not uncommon in patients who have received irradiation to the thyroid gland in the course of treatment for Hodgkin's disease. There is need for continuous reevaluation of the thyroid status in such patients.


Theoretical approaches to the intermolecular nature of water.
A brief review is presented of recent advances in theoretical understanding of liquid water. These advances include detailed quantitative specification of water-molecule interaction potentials, and development of several techniques in statistical mechanics for predicting molecular order in the liquid. Specifically, computer simulation now plays a particularly important rôle. A labile, defective, random network picture for the liquid is the only consistently supported possibility. Implications of this picture for an explanation of hydrophobic solvation and bonding are mentioned. Furthermore, the apparent cooperative restructuring of the random network near the supercolling limit suggests that a dielectric anomaly might exist in this régime. Several areas for future development of water theory are suggested, bearing specifically on phenomena of biological interest.


High alpha-tomatine content in ripe fruit of Andean Lycopersicon esculentum var. cerasiforme: developmental and genetic aspects.
A variant of Lycopersion esculentum var. cerasiforme is described that deviates from the typical form of the entire species, including cultivated tomatoes, in possessing high levels (500-5000 micrograms/g of dry weight) of the steroidal alkaloid alpha-tomatine in its ripe fruits. This biotype is restricted to a tiny enclave in the valley of Río Mayo, Department San Martín, Peru. Among 88 accessions of var. cerasiforme from its present distribution in the Andes, a 90% association was found between high tomatine and bitter flavor; within the Mayo watershed, all samples from the upper drainage had bitterness and high tomatine; the frequency of both traits decreased to low levels toward the lower end. Tomatine therefore probably is the source of bitterness. Throughout L. esculentum tomatine is present at very high concentrations in earliest stages of fruit development, thereafter decreasing rapidly to midperiod, and finally diminishing gradually to near zero at maturity as a result of catabolism to biologically inert compounds, except in the variant described here. High tomatine content does not appear to affect adversely either the natives, among whom the bitter types are popular, or individuals who sampled them in this survey. Genetic determination of high tomatine in ripe fruits is totally recessive and appears to be monogenic with interaction with genes of minor effect. The prevailing pattern of glycoalkaloid synthesis and degradation in development of solanaceous fruits suggests a mechanism to protect against predation prior to ripening but to permit it afterward as a device to promote dispersal. In consideration of the nondegradative nature of the variant, its genetic determination, and very restricted geographic distribution, mutation to this form appears to be a random event of doubtful evolutionary significance.


Exploiting the tumor microenvironment for theranostic imaging.
The integration of chemistry and molecular biology with imaging is providing some of the most exciting opportunities in the treatment of cancer. The field of theranostic imaging, where diagnosis is combined with therapy, is particularly suitable for a disease as complex as cancer, especially now that genomic and proteomic profiling can provide an extensive 'fingerprint' of each tumor. Using this information, theranostic agents can be shaped for personalized treatment to target specific compartments, such as the tumor microenvironment (TME), whilst minimizing damage to normal tissue. These theranostic agents can also be used to target multiple pathways or networks by incorporating multiple small interfering RNAs (siRNAs) within a single agent. A decade ago genetic alterations were the primary focus in cancer research. Now it is apparent that the tumor physiological microenvironment, interactions between cancer cells and stromal cells, such as endothelial cells, fibroblasts and macrophages, the extracellular matrix (ECM), and a host of secreted factors and cytokines, influence progression to metastatic disease, aggressiveness and the response of the disease to treatment. In this review, we outline some of the characteristics of the TME, describe the theranostic agents currently available to target the TME and discuss the unique opportunities the TME provides for the design of novel theranostic agents for cancer therapy.


The Utility of Feticide Procedure to Shorten the Induction-to-Abortion Interval in Medical Abortion.
To evaluate the effect of intracardiac potassium chloride feticide procedure (FP) on the induction-to-abortion (I-to-A) interval for various indications in the termination of pregnancy. Medically indicated abortions between 17 and 28 weeks' gestation were retrospectively evaluated and allocated into 2 groups: Cases with (group 1, n = 58) or without an FP (group 2, n = 60). I-to-A intervals were compared across the groups, considering different baseline parameters and fetal abnormality categories. There were no differences among the groups except in gestational age (GA; mean, 21.2 vs. 19.6 weeks, p = 0.01) in group 1. Overall, the I-to-A interval was shorter (900 ± 233 vs. 1,198 ± 375 min, p = 0.001) and prolonged medical abortion (I-to-A interval > 48 h) was less common (2% vs. 6%, p = 0.03) in group 1. The facilitating effect of FP persisted when indications were categorized as central nervous system, chromosomal, other structural abnormalities, and unclassified conditions. Logistic regression analyses demonstrated the following features to be associated with expulsion of fetus after 24-h: (1) advanced GA (> 24 week; aOR 6.9, 95% CI 3.24-14.72), (2) central nervous system abnormalities (aOR 5.3, 95% CI 2.6-11.4), (3) lack of feticide (aOR 3.67, 95% CI 2.24-10.72). FPs seem to shorten the I-to-A interval and decrease prolonged I-to-A interval rates. This facilitating effect remains unchanged for various medical indications.


Combination warfarin-ASA therapy: which patients should receive it, which patients should not, and why?
Combination warfarin-ASA therapy is currently used in approximately 800,000 patients in North America as long-term treatment for the primary and secondary prevention of atherothrombotic and thromboembolic diseases. Despite a potentially complementary action of anticoagulant and antiplatelet drugs, the use of combination warfarin-ASA therapy is not based on compelling evidence of a net therapeutic benefit, with the exception of patients with a mechanical heart valve. On the other hand, there is more compelling and consistent evidence that combination warfarin-ASA therapy confers a 1.5- to 2.0-fold increased risk for serious bleeding compared with use of warfarin alone. In everyday practice, clinicians should combine the best available evidence with clinical judgment, considering that in most clinical scenarios, clinical practice guideline may not provide clear recommendations for patients who should, and should not, receive combination warfarin-ASA therapy. The objectives of this review are to describe which patients are receiving combined warfarin-aspirin therapy, to summarize the evidence for the therapeutic benefit and harm of combined warfarin-ASA therapy, and to suggest practical guidelines as to which patients should, and should not, receive such treatment.


Anger, hostility, and hospitalizations in patients with heart failure.
Heart failure patients have a high hospitalization rate, and anger and hostility are associated with coronary heart disease morbidity and mortality. Using structural equation modeling, this prospective study assessed the predictive validity of anger and hostility traits for cardiovascular and all-cause rehospitalizations in patients with heart failure. 146 heart failure patients were administered the STAXI and Cook-Medley Hostility Inventory to measure anger, hostility, and their component traits. Hospitalizations were recorded for up to 3 years following baseline. Causes of hospitalizations were categorized as heart failure, total cardiac, noncardiac, and all-cause (sum of cardiac and noncardiac). Measurement models were separately fit for Anger and Hostility, followed by a Confirmatory Factor Analysis to estimate the relationship between the Anger and Hostility constructs. An Anger model consisted of State Anger, Trait Anger, Anger Expression Out, and Anger Expression In, and a Hostility model included Cynicism, Hostile Affect, Aggressive Responding, and Hostile Attribution. The latent construct of Anger did not predict any of the hospitalization outcomes, but Hostility significantly predicted all-cause hospitalizations. Analyses of individual trait components of each of the 2 models indicated that Anger Expression Out predicted all-cause and noncardiac hospitalizations, and Trait Anger predicted noncardiac hospitalizations. None of the individual components of Hostility were related to rehospitalizations or death. The construct of Hostility and several components of Anger are predictive of hospitalizations that were not specific to cardiac causes. Mechanisms common to a variety of health problems, such as self-care and risky health behaviors, may be involved in these associations. (PsycINFO Database Record


Identification of the cytochrome P450 and other enzymes involved in the in vitro oxidative metabolism of a novel antidepressant, Lu AA21004.
1-[2-(2,4-Dimethyl-phenylsulfanyl)-phenyl]-piperazine (Lu AA21004) is a novel antidepressant that is currently in late-stage clinical development for major depressive disorder. In the present study, the metabolism of Lu AA21004 was investigated using human liver microsomes (HLM), human liver S9 fraction, and recombinant enzymes. Lu AA21004 was found in vitro to be oxidized to a 4-hydroxy-phenyl metabolite, a sulfoxide, an N-hydroxylated piperazine, and a benzylic alcohol, which was further oxidized to the corresponding benzoic acid [3-methyl-4-(2-piperazin-1-yl-phenysulfanyl)-benzoic acid (Lu AA34443)]. The formation of the 4-hydroxy-phenyl metabolite was catalyzed by CYP2D6 with some contribution from CYP2C9, whereas the formation of the sulfoxide was mediated by CYP3A4/5 and CYP2A6. CYP2C9 and CYP2C19 were the primary enzymes responsible for formation of the N-hydroxylated metabolite. The benzylic alcohol was formed by CYP2D6 only. The oxidation of the benzylic alcohol to the corresponding benzoic acid of Lu AA21004 was catalyzed by alcohol dehydrogenase and aldehyde dehydrogenase, with some contribution from aldehyde oxidase. CYP2D6 was also capable of catalyzing the formation of the benzoic acid of Lu AA21004; however, its overall contribution to this pathway was negligible. Enzyme kinetic parameters revealed that the rate-limiting step in the formation of the benzoic acid from Lu AA21004 is the formation of the corresponding alcohol. Thus, the intrinsic clearance (V(max)/K(m)) in HLM for metabolism of Lu AA21004 to the benzylic alcohol was 1.13 × 10(-6) l · min(-1) · mg(-1), whereas the subsequent metabolism of the benzylic alcohol to the benzoic acid of Lu AA21004 is characterized by an intrinsic clearance (V(max)/K(m)) in S9 fraction of 922 × 10(-6) l · min(-1) · mg(-1).


Simulation as a means to develop firefighters as emergency care professionals.
Objective. The aim of this study was to evaluate the simulated emergency care performed by firefighters and their perception of simulation as an educational method. Methods. This study had a mixed method with both a quantitative and a qualitative approach. Data were collected by simulation assessment, a questionnaire and written comments. Descriptive analysis was conducted on the quantitative data whereas a qualitative content analysis was conducted on the qualitative data. Finally, a contingent analysis was used where a synthesis configured both the quantitative and the qualitative results into a narrative result. Results. The cognitive workload that firefighters face during simulated emergency care is crucial for learning. In this study, the severity and complexity of the scenarios provided were higher than expected by the firefighters. Clearly stated conditions for the simulation and constructive feedback were considered positive for learning. Patient actors induced realism in the scenario, increasing the experience of stress, in comparison to a manikin. Conclusion. Simulation in a realistic on-scene environment increases firefighters' cognitive ability to critically analyze problems and manage emergency care. Simulation of emergency care developed the firefighters as professionals.


Computer software for performing likelihood tests of pedigree relationship using genetic markers.
Molecular techniques are making ever more genetic markers available for use in parentage assignment, and measures of relatedness. We present a program, Kinship, designed to use likelihood techniques to test for any non-inbred pedigree relationship between pairs of individuals, using single-locus codominant genetic markers. Kinship calculates the likelihood that each pair of individuals in a data set are related by a given pedigree hypothesis, and likelihood ratios for any pair of hypotheses. The program also uses a simulation routine to attach statistical significance to its results.


Adducin: Ca++-dependent association with sites of cell-cell contact.
Adducin is a protein recently purified from erythrocytes and brain that has properties in in vitro assays suggesting a role in assembly of a spectrin-actin lattice. This report describes the localization of adducin to plasma membranes of a variety of tissues and the discovery that adducin is concentrated at sites of cell-cell contact in the epithelial tissues where it is expressed. Adducin in tissues and cultured cells always was observed in association with spectrin and actin, although spectrin and actin were evident in the absence of adducin. In sections of intestinal epithelial cells spectrin was present on all plasma membrane surfaces while adducin was restricted to the lateral cell borders. Adducin also was not detected in association with actin stress fibers in cultured cells. The presence of adducin at cell-cell contact sites of cultured epithelial cells requires extracellular Ca++ and occurs within 15 min of addition of 0.3 mM Ca++. Redistribution of adducin after addition of extracellular Ca++ is independent of formation of desmosomal and adherens junctions since assembly of adducin at contact sites requires lower concentrations of Ca++ and occurs more rapidly than redistribution of desmoplakin or vinculin. Treatment of keratinocytes and MDCK cells with nanomolar concentrations of 12-O-tetradecanoylphorbol-13-acetate (TPA) induces redistribution of adducin away from contact sites. The effect of TPA may be a direct consequence of phosphorylation of adducin, since adducin is phosphorylated in TPA-treated cells and the phosphorylation of adducin occurs before disassembly of adducin from sites of cell-cell contact. Spectrin and adducin are both present in a detergent-insoluble form at cell-cell contact sites of cultured cells. These observations are consistent with the idea that adducin recognizes and associates with specific "receptors" localized at regions of cell-cell contact and promotes assembly of spectrin into a more stable structure, perhaps analogous to the highly organized spectrin-actin network of erythrocyte membranes.


Biallelic inactivation of fumarate hydratase (FH) occurs in nonsyndromic uterine leiomyomas but is rare in other tumors.
Germline mutations in the fumarate hydratase (FH) gene at 1q43 predispose to dominantly inherited cutaneous and uterine leiomyomas, uterine leiomyosarcoma, and papillary renal cell cancer (HLRCC syndrome). To evaluate the role of FH inactivation in sporadic tumorigenesis, we analyzed a series of 299 malignant tumors representing 10 different malignant tumor types for FH mutations. Additionally, 153 uterine leiomyomas from 46 unselected individuals were subjected to and informative in loss of heterozygosity analysis at the FH locus, and the five (3.3%) tumors displaying loss of heterozygosity were subjected to FH mutation analysis. Although mutation search in the 299 malignant tumors was negative, somatic FH mutations were found in two nonsyndromic leiomyomas; a splice site change IVS4 + 3A>G, leading to deletion of exon four, and a missense mutation Ala196Thr. The occurrence of somatic mutations strongly suggests that FH is a true target of the 1q43 deletions. Although uterine leiomyomas are the most common tumors of women, specific inactivating somatic mutations contributing to the formation of nonsyndromic leiomyomas have not been reported previously. Taking into account the apparent risk of uterine leiomyosarcoma associated with FH germline mutations, the finding raises the possibility that also some nonsyndromic leiomyomas may have a genetic profile that is more prone to malignant degeneration. Our data also indicate that somatic FH mutations appear to be limited to tumor types observed in hereditary leiomyomatosis and renal cell cancer.


"The Impact of Mycobacterium tuberculosis Immune Evasion on Protective Immunity: Implications for TB Vaccine Design" - Meeting report.
Tuberculosis (TB) is the major cause of death from infectious diseases around the world, particularly in HIV infected individuals. TB vaccine design and development have been focused on improving Bacille Calmette-Guérin (BCG) and evaluating recombinant and viral vector expressed Mycobacterium tuberculosis (Mtb) proteins, for boosting BCG-primed immunity, but these approaches have not yet yielded significant improvements over the modest effects of BCG in protecting against infection or disease. On March 7-8, 2016, the National Institute of Allergy and Infectious Diseases (NIAID) convened a workshop on "The Impact of Mtb Immune Evasion on Protective Immunity: Implications for TB Vaccine Design" with the goal of defining immune mechanisms that could be targeted through novel research approaches, to inform vaccine design and immune therapeutic interventions for prevention of TB. The workshop addressed early infection events, the impact of Mtb evolution on the development and maintenance of an adaptive immune response, and the factors that influence protection against and progression to active disease. Scientific gaps and areas of study to revitalize and accelerate TB vaccine design were discussed and prioritized. These included a comprehensive evaluation of innate and Mtb-specific adaptive immune responses in the lung at different stages of disease; determining the role of B cells and antibodies (Abs) during Mtb infection; development of better assays to measure Mtb burden following exposure, infection, during latency and after treatment, and approaches to improving current animal models to study Mtb immunogenicity, TB disease and transmission.


Expert Consensus Statements on the Use of Corticosteroids in Non-severe COVID-19.
There is strong evidence for the use of corticosteroid in the management of severe coronavirus disease-2019 (COVID-19). However, there is still uncertainty about the timing of corticosteroids. We undertook a modified Delphi study to develop expert consensus statements on the early identification of a subset of patients from non-severe COVID-19 who may benefit from using corticosteroids. A modified Delphi was conducted with two anonymous surveys between April 30, 2021, and May 3, 2021. An expert panel of 35 experts was selected and invited to participate through e-mail. The consensus was defined as >70% votes in multiple-choice questions (MCQ) on Likert-scale type statements, while strong consensus as >90% votes in MCQ or >50% votes for "very important" on Likert-scale questions in the final round. Twenty experts completed two rounds of the survey. There was strong consensus for the increased work of breathing (95%), a positive six-minute walk test (90%), thorax computed tomography severity score of >14/25 (85%), new-onset organ dysfunction (using clinical or biochemical criteria) (80%), and C-reactive protein >5 times the upper limit of normal (70%) as the criteria for patients' selection. The experts recommended using oral or intravenous (IV) low-dose corticosteroids (the equivalent of 6 mg/day dexamethasone) for 5-10 days and monitoring of oxygen saturation, body temperature, clinical scoring system, blood sugar, and inflammatory markers for any "red-flag" signs. The experts recommended against indiscriminate use of corticosteroids in mild to moderate COVID-19 without the signs of clinical worsening. Oral or IV low-dose corticosteroids (the equivalent of 6 mg/day dexamethasone) for 5-10 days are recommended for patients with features of disease progression based on clinical, biochemical, or radiological criteria after 5 days from symptom onset under close monitoring. How to cite this article: Nasa P, Chaudhry D, Govil D, Daga MK, Jain R, Chhallani AA, et al. Expert Consensus Statements on the Use of Corticosteroids in Non-severe COVID-19. Indian J Crit Care Med 2021;25(11):1280-1285.