Development and Validation of a Quantitative PCR Method for Species Verification and Serogroup Determination of Listeria monocytogenes Isolates.
Listeria monocytogenes (Lm) is one of the leading causes of death because of foodborne illness, affecting the elderly, pregnant women, neonates, and people who are immunocompromised. Serologically, Lm can be classified into 13 serotypes, although only 4 are typically linked with food contamination and illness. Since 2000, a shift in serotypes involved in listeriosis outbreaks has been observed, suggesting that tracking of serotypes could help identify emerging trends. A PCR method developed in 2004 allowed detection of the four major serotypes as molecular serogroups, corresponding to broad phylogenetic groups. In this study, a novel quantitative PCR (qPCR) method was developed that uses two multiplex qPCRs, one to confirm the Listeria genus and Lm species and the second for Lm molecular serogrouping. This method was compared with the U.S. Food and Drug Administration Bacteriological Analytical Manual (BAM) method for Lm and the seroagglutination method, using a 208-strain panel. Comparison of the genus and species qPCR assay with the BAM methods found an equal or slightly higher accuracy for the qPCR method (>98%), compared with the BAM protocol (>96%), when evaluated against independent characterization data. Molecular serogrouping using the qPCR method (96.6%) was more accurate than the seroagglutination assay (75.6%). The qPCR method identified Lm 4bV strains, which could not be resolved using seroagglutination. The qPCR could not identify lineage III and IV serotype 4b strains but did correctly identify 16 of 18 lineage III and IV strains. The qPCR method performed genus identification for the Listeria species Lm, L. innocua, L. welshimeri, L. ivanovii, and L. seeligeri. In addition, the method performed species identification for Lm and classified Lm into six molecular serogroups: 2A, 2B, 2C, 4B, NT, and 4bV. This method provided a rapid and accurate confirmation of Lm and serogroup determinations; furthermore, it could help identify otherwise unlinked strains by enabling whole genome sequencing analysis based on broad phylogeny, independent of other information.


Force sensor in simulated skin and neural model mimic tactile SAI afferent spiking response to ramp and hold stimuli.
The next generation of prosthetic limbs will restore sensory feedback to the nervous system by mimicking how skin mechanoreceptors, innervated by afferents, produce trains of action potentials in response to compressive stimuli. Prior work has addressed building sensors within skin substitutes for robotics, modeling skin mechanics and neural dynamics of mechanotransduction, and predicting response timing of action potentials for vibration. The effort here is unique because it accounts for skin elasticity by measuring force within simulated skin, utilizes few free model parameters for parsimony, and separates parameter fitting and model validation. Additionally, the ramp-and-hold, sustained stimuli used in this work capture the essential features of the everyday task of contacting and holding an object. This systems integration effort computationally replicates the neural firing behavior for a slowly adapting type I (SAI) afferent in its temporally varying response to both intensity and rate of indentation force by combining a physical force sensor, housed in a skin-like substrate, with a mathematical model of neuronal spiking, the leaky integrate-and-fire. Comparison experiments were then conducted using ramp-and-hold stimuli on both the spiking-sensor model and mouse SAI afferents. The model parameters were iteratively fit against recorded SAI interspike intervals (ISI) before validating the model to assess its performance. Model-predicted spike firing compares favorably with that observed for single SAI afferents. As indentation magnitude increases (1.2, 1.3, to 1.4 mm), mean ISI decreases from 98.81 ± 24.73, 54.52 ± 6.94, to 41.11 ± 6.11 ms. Moreover, as rate of ramp-up increases, ISI during ramp-up decreases from 21.85 ± 5.33, 19.98 ± 3.10, to 15.42 ± 2.41 ms. Considering first spikes, the predicted latencies exhibited a decreasing trend as stimulus rate increased, as is observed in afferent recordings. Finally, the SAI afferent's characteristic response of producing irregular ISIs is shown to be controllable via manipulating the output filtering from the sensor or adding stochastic noise. This integrated engineering approach extends prior works focused upon neural dynamics and vibration. Future efforts will perfect measures of performance, such as first spike latency and irregular ISIs, and link the generation of characteristic features within trains of action potentials with current pulse waveforms that stimulate single action potentials at the peripheral afferent.


The technology boom: a new era in obesity management.
As technology continues to develop rapidly, the incidence of obesity also continues to climb at an alarming rate. The increase in available technology is thought to be a contributor in the obesogenic environment, yet at the same time technology can also be used to intervene and improve health and health behaviors. This article reviews the components of effective weight management programs and the novel role that technology, such as SMS, websites, and smartphone apps, is playing to improve the success of such programs. Use of these modern technologies can now allow for individualized treatment recommendations to be delivered to individuals remotely, increased self-monitoring/tracking of health-related data, broader and more rapid dissemination of health information/recommendations, and increased patient-dietician/physician contact. The use of technology in weight management programs results in improved long-term weight management, and in most cases improved cost-effectiveness. Rather than blaming increased food intake and sedentary lifestyle on technology, rapidly developing and innovative technologies should be used to our advantage and deployed to combat the obesity epidemic.


Beyond cigarettes per day. A genome-wide association study of the biomarker carbon monoxide.
The CHRNA5-CHRNA3-CHRNB4 locus is associated with self-reported smoking behavior and also harbors the strongest genetic associations with chronic obstructive pulmonary disease (COPD) and lung cancer. Because the associations with lung disease remain after adjustment for self-reported smoking behaviors, it has been asserted that CHRNA5-CHRNA3-CHRNB4 variants increase COPD and lung cancer susceptibility independently of their effects on smoking. To compare the genetic associations of exhaled carbon monoxide (CO), a biomarker of current cigarette exposure, with self-reported smoking behaviors. A total of 1,521 European American and 247 African American current smokers recruited into smoking cessation studies were assessed for CO at intake before smoking cessation. DNA samples were genotyped using the Illumina Omni2.5 microarray. Genetic associations with CO and smoking behaviors (cigarettes smoked per day, Fagerstrom test for nicotine dependence) were studied. Variants in the CHRNA5-CHRNA3-CHRNB4 locus, including rs16969968, a nonsynonymous variant in CHRNA5, are genomewide association study-significantly associated with CO (β = 2.66; 95% confidence interval [CI], 1.74-3.58; P = 1.65 × 10(-8)), and this association remains strong after adjusting for smoking behavior (β = 2.18; 95% CI, 1.32-3.04; P = 7.47 × 10(-7)). The correlation between CO and cigarettes per day is statistically significantly lower (z = 3.43; P = 6.07 × 10(-4)) in African Americans (r = 0.14; 95% CI, 0.02-0.26; P = 0.003) than in European-Americans (r = 0.36; 95% CI, 0.31-0.40; P = 0.0001). Exhaled CO, a biomarker that is simple to measure, captures aspects of cigarette smoke exposure in current smokers beyond the number of cigarettes smoked per day. Behavioral measures of smoking are therefore insufficient indices of cigarette smoke exposure, suggesting that genetic associations with COPD or lung cancer that persist after adjusting for self-reported smoking behavior may still reflect genetic effects on smoking exposure.


Immunotherapy with chimeric antigen receptors for multiple myeloma.
Chimeric antigen receptors (CARs) are synthetic transmembrane proteins that are used to redirect autologous T cells with a new specificity for antigens on the surface of cancer cells. Impressive results from early phase clinical trials of anti-CD19 CARs for B cell malignancies have generated great enthusiasm for developing this approach for other diseases, particularly hematologic malignancies. Here we review efforts to develop CARs for the treatment of multiple myeloma. Clinical trials are underway investigating CARs against Kappa light chain, CD138, and Lewis Y antigen. CARs against BCMA, CS1, and CD38 are in pre-clinical testing. While initial clinical trials of novel CARs will focus on relapsed/refractory disease, CARs will also likely be studied as a consolidation strategy after response to first-line therapy or in conjunction with autologous hematopoietic stem cell transplantation.


The effects of estrone (Ogen) on spinal bone density of postmenopausal women.
The effects of cyclical treatment with estrone sulfate (0.3, 0.625, or 1.25 mg), plus calcium carbonate, on spinal trabecular bone density were compared with placebo in 120 postmenopausal women in this 2-year, multicenter, double-blind study. While the placebo and 0.3-mg treatment groups lost bone density (-3.6% and -5.1%), the 0.625- and 1.25-mg treatment groups experienced no significant change from baseline at 24 months (-0.8% and +0.7%). The 1.25-mg treatment group was significantly different from the placebo group at 12, 18, and 24 months. Although the 0.625-mg treatment group was significantly different from the placebo group only at 18 months, the data suggest that 0.625 and 1.25 mg of estrone sulfate had different effects than placebo and 0.3 mg of estrone sulfate and, given with supplemental calcium, are effective doses for the prevention of spinal bone loss.


Impact of hepatitis B virus (HBV) x gene mutations on hepatocellular carcinoma development in chronic HBV infection.
The hepatitis B virus (HBV) PreS mutations C1653T, T1753V, and A1762T/G1764A were reported as a strong risk factor of hepatocellular carcinoma (HCC) in a meta-analysis. HBV core promoter overlaps partially with HBx coding sequence, so the nucleotide 1762 and 1764 mutations induce HBV X protein (HBx) 130 and 131 substitutions. We sought to elucidate the impact of HBx mutations on HCC development. Chronically HBV-infected patients were enrolled in this study: 42 chronic hepatitis B (CHB) patients, 23 liver cirrhosis (LC) patients, and 31 HCC patients. Direct sequencing showed HBx131, HBx130, HBx5, HBx94, and HBx38 amino acid mutations were common in HCC patients. Of various mutations, HBx130+HBx131 (double) mutations and HBx5+HBx130+HBx131 (triple) mutations were significantly high in HCC patients. Double and triple mutations increased the risk for HCC by 3.75-fold (95% confidence interval [CI] = 1.101 to 12.768, P = 0.033) and 5.34-fold (95% CI = 1.65 to 17.309, P = 0.005), respectively, when HCC patients were compared to CHB patients. Functionally, there were significantly higher levels of NF-κB activity in cells with the HBx5 mutant and with the double mutants than that of wild-type cells and the triple-mutant cells. The triple mutation did not increase NF-κB activity. Other regulatory pathways seem to exist for NF-κB activation. In conclusion, a specific HBx mutation may contribute to HCC development by activating NF-κB activity. The HBx5 mutation in genotype C2 HBV appears to be a risk factor for the development of HCC and may be used to predict the clinical outcomes of patients with chronic HBV infection.


Parenting Behaviors and Family Conflict as Predictors of Adolescent Sleep and Bedtime Media Use.
Inadequate sleep in adolescents has been linked to an increase in screen-based media use, especially at bedtime. Parents can play a critical role in regulating adolescent media use and promoting healthy sleep, yet few studies have evaluated parental effects on these outcomes. This study examined the effects of general and media-specific parenting behaviors and family conflict on adolescent sleep outcomes, both directly and indirectly through bedtime media use. Data were collected from 345 middle-schoolers (Mage = 12.65 ± 0.67 years; 47% female; 59% White) at two time points, six months apart. The findings revealed that parental involvement had a significant positive effect on sleep duration that was mediated by bedtime media use. Family conflict had a direct positive effect on daytime sleepiness. Adolescent sleep interventions could benefit from a parenting component focused on positive involvement and fostering a family climate conducive to sleep.


Quantifying and analyzing the network basis of genetic complexity.
Genotype-to-phenotype maps exhibit complexity. This genetic complexity is mentioned frequently in the literature, but a consistent and quantitative definition is lacking. Here, we derive such a definition and investigate its consequences for model genetic systems. The definition equates genetic complexity with a surplus of genotypic diversity over phenotypic diversity. Applying this definition to ensembles of Boolean network models, we found that the in-degree distribution and the number of periodic attractors produced determine the relative complexity of different topology classes. We found evidence that networks that are difficult to control, or that exhibit a hierarchical structure, are genetically complex. We analyzed the complexity of the cell cycle network of Sacchoromyces cerevisiae and pinpointed genes and interactions that are most important for its high genetic complexity. The rigorous definition of genetic complexity is a tool for unraveling the structure and properties of genotype-to-phenotype maps by enabling the quantitative comparison of the relative complexities of different genetic systems. The definition also allows the identification of specific network elements and subnetworks that have the greatest effects on genetic complexity. Moreover, it suggests ways to engineer biological systems with desired genetic properties.


Intensity of hemorrhage following tonsillectomy.
Although post-tonsillectomy hemorrhage is one of the most frequent and potentially life-threatening complications, there is no generally accepted classification of post-operative bleeding intensity. The aim of this study was to evaluate the intensity of post-tonsillectomy hemorrhage according to the five-grade classification. A total of 408 consecutive patients, aged 2 to 54 years, undergoing elective tonsillectomy, with (n = 261) or without (n = 147) adenoidectomy, were included in this prospective study. Tonsillectomy was performed under general anesthesia using standard technique of cold dissection with a snare. Any bleeding event was recorded. The severity of post-operative hemorrhage was classified in five grades. In 11 (2.70%) of the patients grade 1 hemorrhage following tonsillectomy occurred, 4 (0.98%) had grade 2 and 2 (0.49%) of the patients had grade 3 post-operative bleeding. Grades 4 and 5 were not recorded, and no patient received a blood transfusion. Post-tonsillectomy hemorrhage can be expected in a small number of patients undergoing tonsillectomy. Hemorrhage is mostly primary and rarely requires treatment under endotracheal anesthesia and blood transfusion.


[Imaging techniques in modern trauma diagnostics].
Modern trauma room management requires interdisciplinary teamwork and synchronous communication between a team of anaesthesists, surgeons and radiologists. As the length of stay in the trauma room influences morbidity and mortality of a severely injured person, optimizing time is one of the main targets. With the direct involvement of modern imaging techniques the injuries caused by trauma should be detected within a very short period of time in order to enable a priority-orientated treatment. Radiology influences structure and process quality, management and development of trauma room algorithms regarding the use of imaging techniques. For the individual case interventional therapy methods can be added. Based on current data and on the Frankfurt experience the current diagnostic concepts of trauma diagnostics are presented.


Toward a brain-computer interface for Alzheimer's disease patients by combining classical conditioning and brain state classification.
Brain-computer interfaces (BCIs) provide alternative methods for communicating and acting on the world, since messages or commands are conveyed from the brain to an external device without using the normal output pathways of peripheral nerves and muscles. Alzheimer's disease (AD) patients in the most advanced stages, who have lost the ability to communicate verbally, could benefit from a BCI that may allow them to convey basic thoughts (e.g., "yes" and "no") and emotions. There is currently no report of such research, mostly because the cognitive deficits in AD patients pose serious limitations to the use of traditional BCIs, which are normally based on instrumental learning and require users to self-regulate their brain activation. Recent studies suggest that not only self-regulated brain signals, but also involuntary signals, for instance related to emotional states, may provide useful information about the user, opening up the path for so-called "affective BCIs". These interfaces do not necessarily require users to actively perform a cognitive task, and may therefore be used with patients who are cognitively challenged. In the present hypothesis paper, we propose a paradigm shift from instrumental learning to classical conditioning, with the aim of discriminating "yes" and "no" thoughts after associating them to positive and negative emotional stimuli respectively. This would represent a first step in the development of a BCI that could be used by AD patients, lending a new direction not only for communication, but also for rehabilitation and diagnosis.


Crystal structure of Tapes japonica Lysozyme with substrate analogue: structural basis of the catalytic mechanism and manifestation of its chitinase activity accompanied by quaternary structural change.
Tapes japonica lysozyme (TJL) is classified as a member of the recently established i-type lysozyme family. In this study, we solved the crystal structure of TJL complexed with a trimer of N-acetylglucosamine to 1.6A resolution. Based on structure and mutation analyses, we demonstrated that Glu-18 and Asp-30 are the catalytic residues of TJL. Furthermore, the present findings suggest that the catalytic mechanism of TJL is a retaining mechanism that proceeds through a covalent sugar-enzyme intermediate. On the other hand, the quaternary structure in the crystal revealed a dimer formed by the electrostatic interactions of catalytic residues (Glu-18 and Asp-30) in one molecule with the positive residues at the C terminus in helix 6 of the other molecule. Gel chromatography analysis revealed that the TJL dimer remained intact under low salt conditions but that it dissociated to TJL monomers under high salt conditions. With increasing salt concentrations, the chitinase activity of TJL dramatically increased. Therefore, this study provides novel evidence that the lysozyme activity of TJL is modulated by its quaternary structure.


[Visceral artery aneurysms].
The purpose of this article is to review the etiology, clinical presentation and treatment options of visceral artery aneurysms (VAA) on the basis of our experiences. Visceral artery aneurysms are uncommon lesions with a frequency of 0,1-0,2 % in autopsy statistics. In fact many visceral artery aneurysms still present first with a rupture (22 %) and despite emergency laparotomies the mortality rate is about 8,5 %. The course of disease often is disastrous due to rupture of the aneurysms or thromboembolic complications, emphasizing the importance to be aware of this differential diagnosis of abdominal pain. This article covers 9 patients with VAA. 3 patients each revealed an a. lienalis aneurysm and a. gastrica aneurysm, resp. In the other 3 patients an aneurysm of the a. gastroepiploica, the a. pancreatico-duodenalis and the a. mesenterica superior resp. was proven. In 8 of 9 patients a surgical therapy of the VAA took place. Only 2 patients (22 %) were interventionally treated. 1 patient deceased due to postoperative hemorrhage. Both the surgical and the radiological intervention therapy are available for treatment of the VAA. The decision on the choice of the therapeutic procedure should be made on an individual basis.


[The pathomorphological characteristics of duodenal peptic ulcer in the victims of the accident at the Chernobyl Atomic Electric Power Station: changes in the duodenal mucosa outside of the defect].
A study was made of duodenobiopsies from patients with duodenal ulcer, 62 of whom took part in the elimination of the effects of the Chernobyl Nuclear Power Plant accident, 33 have been resident in the territories affected by radioactive contamination, and 36 were a group of nosological control. In the ChNPP accident victims, the inflammatory process in chronic duodenitis tended to be more pronounced and active, with the atrophic changes in the mucous membrane being more commonly seen. Incorporation of radionuclides and external irradiation in a dose exceeding 25 sGy causes most profound disturbances in regeneration of epithelium entailing metaplasia and dysplasia of gastric epithelium in duodenum, which facts heighten the risk for neoplasmogenesis.


Piecing together numerical language: children's use of default units in early counting and quantification.
When asked to 'find three forks', adult speakers of English use the noun 'fork' to identify units for counting. However, when number words (e.g. three) and quantifiers (e.g. more, every) are used with unfamiliar words ('Give me three blickets') noun-specific conceptual criteria are unavailable for picking out units. This poses a problem for young children learning language, who begin to use quantifiers and number words by age 2, despite knowing a relatively small number of nouns. Without knowing how individual nouns pick out units of quantification--e.g. what counts as a blicket--how could children decide whether there are three blickets or four? Three experiments suggest that children might solve this problem by assigning 'default units' of quantification to number words, quantifiers, and number morphology. When shown objects that are broken into arbitrary pieces, 4-year-olds in Experiment 1 treated pieces as units when counting, interpreting quantifiers, and when using singular-plural morphology. Experiment 2 found that although children treat object-hood as sufficient for quantification, it is not necessary. Also sufficient for individuation are the criteria provided by known nouns. When two nameable things were glued together (e.g. two cups), children counted the glued things as two. However, when two arbitrary pieces of an object were put together (e.g. two parts of a ball), children counted them as one, even if they had previously counted the pieces as two. Experiment 3 found that when the pieces of broken things were nameable (e.g. wheels of a bicycle), 4-year-olds did not include them in counts of whole objects (e.g. bicycles). We discuss the role of default units in early language acquisition, their origin in acquisition, and how children eventually acquire an adult semantics identifying units of quantification.


Assessment and optimization of cell engraftment after transplantation into the heart.
Myocardial regeneration using stem and progenitor cell transplantation in the injured heart has recently become a major goal in the treatment of cardiac disease. Experimental studies and clinical applications have generally been encouraging, although the functional benefits that have been attained clinically are modest and inconsistent. Low cell retention and engraftment after myocardial delivery is a key factor limiting the successful application of cell therapy, irrespective of the type of cell or the delivery method. To improve engraftment, accurate methods for tracking cell fate and quantifying cell survival need to be applied. Several laboratory techniques (histological methods, real-time quantitative polymerase chain reaction, radiolabeling) have provided invaluable information about cell engraftment. In vivo imaging (nuclear medicine modalities, bioluminescence, and MRI) has the potential to provide quantitative information noninvasively, enabling longitudinal assessment of cell fate. In the present review, we present several available methods for assessing cell engraftment, and we critically discuss their strengths and limitations. In addition to providing insights about the mechanisms mediating cell loss after transplantation, these methods can evaluate techniques for augmenting engraftment, such as tissue engineering approaches, preconditioning, and genetic modification, allowing optimization of cell therapies.


Role of lipid A acylation in Yersinia enterocolitica virulence.
Yersinia enterocolitica is an important human pathogen. Y. enterocolitica must adapt to the host environment, and temperature is an important cue regulating the expression of most Yersinia virulence factors. Here, we report that Y. enterocolitica 8081 serotype O:8 synthesized tetra-acylated lipid A at 37 degrees C but that hexa-acylated lipid A predominated at 21 degrees C. By mass spectrometry and genetic methods, we have shown that the Y. enterocolitica msbB, htrB, and lpxP homologues encode the acyltransferases responsible for the addition of C(12), C(14) and C(16:1), respectively, to lipid A. The expression levels of the acyltransferases were temperature regulated. Levels of expression of msbB and lpxP were higher at 21 degrees C than at 37 degrees C, whereas the level of expression of htrB was higher at 37 degrees C. At 21 degrees C, an lpxP mutant was the strain most susceptible to polymyxin B, whereas at 37 degrees C, an htrB mutant was the most susceptible. We present evidence that the lipid A acylation status affects the expression of Yersinia virulence factors. Thus, expression of flhDC, the flagellar master regulatory operon, was downregulated in msbB and lpxP mutants, with a concomitant decrease in motility. Expression of the phospholipase yplA was also downregulated in both mutants. inv expression was downregulated in msbB and htrB mutants, and consistent with this finding, invasion of HeLa cells was diminished. However, the expression of rovA, the positive regulator of inv, was not affected in the mutants. The levels of pYV-encoded virulence factors Yops and YadA in the acyltransferase mutants were not affected. Finally, we show that only the htrB mutant was attenuated in vivo.


Deformable and durable phantoms with controlled density of scatterers.
We have developed deformable and durable optical tissue phantoms with a simple and well-defined microstructure including a novel combination of scatterers and a matrix material. These were developed for speckle and elastography investigations in optical coherence tomography, but should prove useful in many other fields. We present in detail the fabrication process which involves embedding silica microspheres in a silicone matrix. We also characterize the resulting phantoms with scanning electron microscopy and optical measurements. To our knowledge, no such phantoms were proposed in the literature before. Our technique has a wide range of applicability and could also be adapted to fabricate phantoms with various optical and mechanical properties.


Chemical modification of a functional arginine residue in diadenosine 5',5'''-P1,P4-tetraphosphate (Ap4A) phosphorylase I from Saccharomyces cerevisiae.
Phenylglyoxal, a reagent with high specificity for arginine residues, inactivated Ap4A phosphorylase I from Saccharomyces cerevisiae in a pseudo-first-order manner. The second-order rate constant was 11.5 +/- 2.5 M-1 min-1. The loss of activity was a linear function of the incorporation of [7-14C]phenylglyoxal. The incorporation of 1.9 +/- 0.4 mol of phenylglyoxal/mol of enzyme accounted for complete loss of activity. The specificity of inactivation by phenylglyoxal was tested in the presence of ApnA (n = 2-6), ADP, ATP and Pi. The substrates, Ap4A, Ap5A and Pi protected the enzyme against inactivation, but Ap2A, Ap3A and Ap6A did not. Ap4A, Ap5A and Pi reduced the rate of inactivation by about 70%, 60% and 37% respectively. The Ap4A phosphorolysis products, ADP and ATP, also partially protected the enzyme against inactivation by phenylglyoxal. Thus Ap4A phosphorylase I probably contains an arginine residue in the binding site for Ap4A.


[Primary structure of the potato virus X genome: the region preceding the capsid protein cistron].
DNA copies of the potato virus X (PVX) RNA corresponding to 2300 nucleotides at the 3'-end have been cloned. The cloned cDNA copies containing the nucleotides 445-1280 from the 3'-end have been sequenced. The 5'-terminal region of the PVX coat protein gene corresponds to residues 445-786 from the 3'-end. The amino acid sequences of two more open reading frames (ORF) have been deduced from the nucleotide sequence. The potential translation products of these ORF's would correspond to the nonstructural viral proteins. We have located the ORF1 within the region of residues 799-1009 preceding the coat protein cistron. The tentative protein is composed of 70 amino acids and has an aminoterminal segment which is markedly hydrophobic. ORF2 in the PVX sequence ends with UAG at nucleotides 942-944 and extends to the 5'-terminus for additional 340 nucleotides. The distant sequence homology exists between a carboxyterminal portion of PVX ORF2 and that of the nonstructural "30 K-proteins" of the plant tobamoviruses.


Efficient implementation for spherical flux computation and its application to vascular segmentation.
Spherical flux is the flux inside a spherical region, and it is very useful in the analysis of tubular structures in magnetic resonance angiography and computed tomographic angiography. The conventional approach is to estimate the spherical flux in the spatial domain. Its running time depends on the sphere radius quadratically, which leads to very slow spherical flux computation when the sphere size is large. This paper proposes a more efficient implementation for spherical flux computation in the Fourier domain. Our implementation is based on the reformulation of the spherical flux calculation using the divergence theorem, spherical step function, and the convolution operation. With this reformulation, most of the calculations are performed in the Fourier domain. We show how to select the frequency subband so that the computation accuracy can be maintained. It is experimentally demonstrated that, using the synthetic and clinical phase contrast magnetic resonance angiographic volumes, our implementation is more computationally efficient than the conventional spatial implementation. The accuracies of our implementation and that of the conventional spatial implementation are comparable. Finally, the proposed implementation can definitely benefit the computation of the multiscale spherical flux with a set of radii because, unlike the conventional spatial implementation, the time complexity of the proposed implementation does not depend on the sphere radius.


COVID-19 optimal vaccination policies: A modeling study on efficacy, natural and vaccine-induced immunity responses.
About a year into the pandemic, COVID-19 accumulates more than two million deaths worldwide. Despite non-pharmaceutical interventions such as social distance, mask-wearing, and restrictive lockdown, the daily confirmed cases remain growing. Vaccine developments from Pfizer, Moderna, and Gamaleya Institute reach more than 90% efficacy and sustain the vaccination campaigns in multiple countries. However, natural and vaccine-induced immunity responses remain poorly understood. There are great expectations, but the new SARS-CoV-2 variants demand to inquire if the vaccines will be highly protective or induce permanent immunity. Further, in the first quarter of 2021, vaccine supply is scarce. Consequently, some countries that are applying the Pfizer vaccine will delay its second required dose. Likewise, logistic supply, economic and political implications impose a set of grand challenges to develop vaccination policies. Therefore, health decision-makers require tools to evaluate hypothetical scenarios and evaluate admissible responses. Following some of the WHO-SAGE recommendations, we formulate an optimal control problem with mixed constraints to describe vaccination schedules. Our solution identifies vaccination policies that minimize the burden of COVID-19 quantified by the number of disability-adjusted years of life lost. These optimal policies ensure the vaccination coverage of a prescribed population fraction in a given time horizon and preserve hospitalization occupancy below a risk level. We explore "via simulation" plausible scenarios regarding efficacy, coverage, vaccine-induced, and natural immunity. Our simulations suggest that response regarding vaccine-induced immunity and reinfection periods would play a dominant role in mitigating COVID-19.


Nociceptin/orphanin FQ-induced inhibition of delayed rectifier potassium currents by calcium/calmodulin-dependent protein kinase type II.
The neuropeptide nociceptin/orphanin FQ (N/OFQ) has been shown to inhibit delayed rectifier potassium current (IK) in acutely dissociated rat parietal cortical neurons. However, the detailed mechanism of N/OFQ-induced inhibition on IK is not clear. This study is the first to explore an involvement of calcium/calmodulin (CaM)-dependent protein kinase type II (CaMKII) in mediating N/OFQ-induced responses. Utilizing pharmacological inhibitors of CaM and CaMKII, we have investigated the contribution of CaMKII in N/OFQ-induced effects using the whole-cell patch-clamp technique. In whole-cell voltage clamp, W-7 (100 μM), an antagonist of CaM, as well as KN-62, an inhibitor of CaMKII activity, attenuated the inhibitory effects of N/OFQ on IK. Activation and inactivation analysis indicated that the kinetics of IK were altered by N/OFQ, with decreased activation and promoted inactivation of IK. W-7 and KN-62 (10 μM) partly abolished the activation and inactivation curves shift of IK induced by N/OFQ. These findings show that CaMKII plays a critical role in N/OFQ-induced inhibition of IK in acutely dissociated rat parietal cortical neurons.


The Genetics of Axon Guidance and Axon Regeneration in Caenorhabditis elegans.
The correct wiring of neuronal circuits depends on outgrowth and guidance of neuronal processes during development. In the past two decades, great progress has been made in understanding the molecular basis of axon outgrowth and guidance. Genetic analysis in Caenorhabditis elegans has played a key role in elucidating conserved pathways regulating axon guidance, including Netrin signaling, the slit Slit/Robo pathway, Wnt signaling, and others. Axon guidance factors were first identified by screens for mutations affecting animal behavior, and by direct visual screens for axon guidance defects. Genetic analysis of these pathways has revealed the complex and combinatorial nature of guidance cues, and has delineated how cues guide growth cones via receptor activity and cytoskeletal rearrangement. Several axon guidance pathways also affect directed migrations of non-neuronal cells in C. elegans, with implications for normal and pathological cell migrations in situations such as tumor metastasis. The small number of neurons and highly stereotyped axonal architecture of the C. elegans nervous system allow analysis of axon guidance at the level of single identified axons, and permit in vivo tests of prevailing models of axon guidance. C. elegans axons also have a robust capacity to undergo regenerative regrowth after precise laser injury (axotomy). Although such axon regrowth shares some similarities with developmental axon outgrowth, screens for regrowth mutants have revealed regeneration-specific pathways and factors that were not identified in developmental screens. Several areas remain poorly understood, including how major axon tracts are formed in the embryo, and the function of axon regeneration in the natural environment.


A new FRAX model for Brazil.
Fracture probabilities derived from the original FRAX model for Brazil were compared to those from an updated model based on more recent regional estimates of the incidence of hip fracture. Fracture probabilities were consistently lower in the updated FRAX model. Despite large differences between models, differences in the rank order of fracture probabilities were minimal. Recent epidemiological data indicate that the risk of hip fracture in Brazil is lower than that used to create the original FRAX model. This paper describes the epidemiology of hip fracture in Brazil and the synthesis of an updated FRAX model with the aim of comparing this new model with the original model. Hip fracture rates from three cities in three regions were combined, weighted by the population of each region. For other major fractures, incidence rates for Brazil were estimated using Swedish ratios for hip to other major osteoporotic fracture (humerus, forearm or clinical vertebral fractures). Mortality estimates were taken from the UN. Compared to the original FRAX model, the updated model gave lower 10-year fracture probabilities in men and women at all ages. Notwithstanding, there was a very close correlation in fracture probabilities between the original and updated models (r > 0.99) so that the revisions had little impact on the rank order of risk. The disparities between the original and updated FRAX models indicate the importance of updating country-specific FRAX models with the advent of significant changes in fracture epidemiology.


Nebivolol ameliorated kidney damage in Zucker diabetic fatty rats by regulation of oxidative stress/NO pathway: Comparison with captopril.
The aim was to evaluate the effects and mechanisms of nebivolol on renal damage in Zucker diabetic fatty (ZDF) rats, in comparison with those of atenolol and captopril. Animals were divided into: control lean Zucker rats, ZDF rats, ZDF rats orally treated with nebivolol (10 mg/kg), atenolol (100 mg/kg) or captopril (40 mg/kg) for 6 months. Systolic blood pressure (SBP), blood glucose, kidney structure and function, plasma and kidney levels of nitric oxide (NO) and asymmetric dimethylarginine (ADMA), and oxidant status were evaluated. Kidney expressions of adenosine monophosphate-activated protein kinase (AMPK), NADPH oxidase (NOX) isoforms 2 and 4 and subunit p22phox , nitric oxide synthase (NOS) isoforms, endothelial NOS (eNOS) uncoupling, protein arginine N-methyltransferase (PRMT) 1, and dimethylarginine dimethylaminohydrolase (DDAH) 1 and 2 were tested. All drugs induced a similar control of SBP. Nebivolol did not affect the increased plasma glucose. Unlike atenolol, nebivolol prevented the decrease in plasma insulin, and, like captopril, it reduced plasma lipid contents. Nebivolol ameliorated, to a greater extent than captopril, damages to renal structure and function, which were associated with an improvement in interlobular artery dysfunction. Nebivolol elevated kidney phosphorylation of AMPK, attenuated NOX4 and p22phox expression and oxidative stress marker levels. Nebivolol increased plasma and renal NO, enhanced expressions of eNOS, p-eNOS and neuronal NOS, and suppressed eNOS uncoupling and inducible NOS expression. High ADMA in plasma and kidney were decreased by nebivolol through increasing DDAH2 and decreasing protein arginine N-methyltransferase 1. Long-term treatment of nebivolol ameliorated diabetic nephropathy, at least in part, via regulation of renal oxidative stress/NO pathway.


Target-independent diversification and target-specific projection of chemically defined retinal ganglion cell subsets.
In diverse vertebrate species, defined subsets of retinal ganglion cells (RGCs, the neurons that project from retina to brain) are distinguishable on the basis of their dendritic morphology, physiological properties, neurotransmitter content and synaptic targets. Little is known about when this diversity arises, whether diversification requires target-derived signals, and how subtype-specific projection patterns are established. Here, we have used markers for two chemically defined RGC subsets in chick retina to address these issues. Antibodies to substance P (SP) and the nicotine acetylcholine receptor (AChR) beta 2 subunit label two small ( < 10%), mutually exclusive groups of RGCs in mature retina. SP and AChRs accumulate in distinct RGCs before retinotectal synapses have formed. Moreover, both populations of RGCs form in retinae that develop following tectal ablation or transplantation to the coelomic cavity. Thus, RGC subsets acquire distinct neurotransmitter phenotypes in the absence of extraretinal cues. In the mature optic tectum, SP- and AChR-positive RGC axonal arbors are confined to distinct retinorecipient (synaptic) laminae. In the developing tectum, SP- and AChR-positive axons are initially intermingled in a superficial fiber layer, but then enter and arborize in appropriate laminae soon after those laminae form. Importantly, SP-positive axons, which synapse in a superficial lamina, never extend into the deeper, AChR-positive lamina. Tectal interneurons rich in SP receptors are concentrated in the lamina to which SP-positive RGC axons project, and a set of cholinergic (choline acetyltransferase-positive) tectal projection neurons elaborate dendrites in the lamina to which AChR-positive RGC axons project. These populations of tectal neurons, which are likely targets of the RGC subsets, form in tecta that develop following enucleation. Thus, RGCs and their targets can diversify in each others absence. Accordingly, we propose that the lamina-selective connectivity we observe reflects the presence of complementary cues on RGC subsets and their laminar targets.


Arginine deficiency caused by myeloid cells: importance, identification and treatment.
Dietary arginine supplementation has been suggested as a means of improving T lymphocyte function and has found its greatest clinical utility in patients undergoing elective surgery. In other illnesses, arginine supplementation is controversial. Breakthroughs in understanding arginine metabolism have led to the identification of myeloid cells that express arginase 1, causing significant depletion of arginine - an essential amino acid for normal T lymphocyte function. Hence, myeloid cells expressing arginase 1 are also known as myeloid-derived suppressor cells. This chapter discusses the hypothesis that arginine replacement therapy may be necessary in arginine deficiency states.


Effects of cortisol and salinity challenge on water balance in developing larvae of tilapia (Oreochromis mossambicus).
Effects of exogenous cortisol on drinking rate and water content in developing larvae of tilapia (Oreochromis mossambicus) were examined. Both freshwater- and seawater-adapted larvae showed increases in drinking rates with development. Drinking rates of seawater-adapted larvae were about four- to ninefold higher than those of freshwater-adapted larvae from day 2 to day 5 after hatching. Seawater-adapted larvae showed declines in drinking rate and water content at 4 and 14 h, respectively, after immersion in 10 mg L(-1) cortisol. In the case of freshwater-adapted larvae, the drinking rate decreased after 8 h of cortisol immersion, while the water content did not show a significant change even after 32 h of cortisol immersion. In a subsequent experiment of transfer from freshwater to 20 ppt (parts per thousand, salinity) seawater, immersion in 10 mg L(-1) cortisol for 8-24 h enhanced the drinking rate in larvae at 4 h after transfer, but no significant difference was found in water contents between cortisol-treated and control groups following transfer. These results suggest that cortisol is involved in the regulation of drinking activity in developing tilapia larvae.


Exercise training in aging and diseases.
Sedentary lifestyle along with high blood pressure, abnormal values for blood lipids, smoking, and obesity are recognized risk factors for cardiovascular diseases and for many other chronic diseases, such as diabetes, osteoporosis, breast and colon cancer. Several studies conducted on large cohort of individuals have documented the protective effects of physical activity for both vascular and nonvascular syndromes. Exercise training is an integral part of cardiac rehabilitation, a complex therapeutic approach, effective both in young and elderly patients. Despite the number of evidences underling the benefits associated with physical fitness, the cardiac rehabilitation is still an underused medical resource. The molecular mechanism behind physical activity protective effect is presently unresolved, and further studies are also needed to establish the best protocol in terms of specificity, volume and duration of the training.


Automated Quantitative Stress Perfusion Cardiac Magnetic Resonance in Pediatric Patients.
Background: Myocardial ischemia occurs in pediatrics, as a result of both congenital and acquired heart diseases, and can lead to further adverse cardiac events if untreated. The aim of this work is to assess the feasibility of fully automated, high resolution, quantitative stress myocardial perfusion cardiac magnetic resonance (CMR) in a cohort of pediatric patients and to evaluate its agreement with the coronary anatomical status of the patients. Methods: Fourteen pediatric patients, with 16 scans, who underwent dual-bolus stress perfusion CMR were retrospectively analyzed. All patients also had anatomical coronary assessment with either CMR, CT, or X-ray angiography. The perfusion CMR images were automatically processed and quantified using an analysis pipeline previously developed in adults. Results: Automated perfusion quantification was successful in 15/16 cases. The coronary perfusion territories supplied by vessels affected by a medium/large aneurysm or stenosis (according to the AHA guidelines), induced by Kawasaki disease, an anomalous origin, or interarterial course had significantly reduced myocardial blood flow (MBF) (median (interquartile range), 1.26 (1.05, 1.67) ml/min/g) as compared to territories supplied by unaffected coronaries [2.57 (2.02, 2.69) ml/min/g, p < 0.001] and territories supplied by vessels with a small aneurysm [2.52 (2.45, 2.83) ml/min/g, p = 0.002]. Conclusion: Automatic CMR-derived MBF quantification is feasible in pediatric patients, and the technology could be potentially used for objective non-invasive assessment of ischemia in children with congenital and acquired heart diseases.


EWS-FLI1 modulated alternative splicing of ARID1A reveals novel oncogenic function through the BAF complex.
Connections between epigenetic reprogramming and transcription or splicing create novel mechanistic networks that can be targeted with tailored therapies. Multiple subunits of the chromatin remodeling BAF complex, including ARID1A, play a role in oncogenesis, either as tumor suppressors or oncogenes. Recent work demonstrated that EWS-FLI1, the oncogenic driver of Ewing sarcoma (ES), plays a role in chromatin regulation through interactions with the BAF complex. However, the specific BAF subunits that interact with EWS-FLI1 and the precise role of the BAF complex in ES oncogenesis remain unknown. In addition to regulating transcription, EWS-FLI1 also alters the splicing of many mRNA isoforms, but the role of splicing modulation in ES oncogenesis is not well understood. We have identified a direct connection between the EWS-FLI1 protein and ARID1A isoform protein variant ARID1A-L. We demonstrate here that ARID1A-L is critical for ES maintenance and supports oncogenic transformation. We further report a novel feed-forward cycle in which EWS-FLI1 leads to preferential splicing of ARID1A-L, promoting ES growth, and ARID1A-L reciprocally promotes EWS-FLI1 protein stability. Dissecting this interaction may lead to improved cancer-specific drug targeting.


Improved Macrophage Enrichment from Mouse Skeletal Muscle.
Macrophages are a heterogeneous class of innate immune cells that offer a primary line of defense to the body by phagocytizing pathogens, digesting them, and presenting the antigens to T and B cells to initiate adaptive immunity. Through specialized pro-inflammatory or anti-inflammatory activities, macrophages also directly contribute to the clearance of infections and the repair of tissue injury. Macrophages are distributed throughout the body and largely carry out tissue-specific functions. In skeletal muscle, macrophages regulate tissue repair and regeneration; however, the characteristics of these macrophages are not yet fully understood, and their involvement in skeletal muscle aging remains to be elucidated. To investigate these functions, it is critical to efficiently isolate macrophages from skeletal muscle with sufficient purity and yield for various downstream analyses. However, methods to prepare enriched skeletal muscle macrophages are scarce. Here, we describe in detail an optimized method to isolate skeletal muscle macrophages from mice. This method has allowed the isolation of CD45 + /CD11b + macrophage-enriched cells from young and old mice, which can be further used for flow cytometric analysis, fluorescence-activated cell sorting (FACS), and single-cell RNA sequencing. This protocol was validated in: eLife (2022), DOI: 10.7554/eLife.77974.


[How many patients with Creutzfeldt-Jakob disease are there in Russia?]
The article presents epidemiological data on the prevalence of Creutzfeldt-Jakob disease in the world and modern techniques for the rapid accurate diagnosis. The author discusses a national approach to this subject.


[Clinical management of primary headaches: interdisciplinary guidelines].
Primary headaches, whose aetiologies cannot be identified by current diagnostic methods, represent the most frequent headaches and must be distinguished from secondary or symptomatic headaches, e.g. related to traumatic, vascular, inflammatory or neoplastic processes. Only an appropriate treatment will meet the expectations of patients, for who even a mild headache may generate important fears and handicaps. Medical history and physical examination form the basis for targeting the right diagnosis before any biological or radiological testing. The article presents guidelines implemented at the Geneva University Hospitals by the Pain Network of the institution. Therapeutic modalities and recommendations concerning the need for specialized assessments are presented to provide a framework in both emergency situation and long-term care.


The synergistic role of carbon quantum dots for the improved photocatalytic performance of Bi2MoO6.
Novel carbon quantum dot (CQD) modified Bi2MoO6 photocatalysts were prepared via a facile hydrothermal process. The CQD modified Bi2MoO6 materials were characterized by multiple techniques. The CQDs with the average size of about 7 nm were distributed on the surface of Bi2MoO6 nanosheets. The photocatalytic activity of as-prepared CQD modified Bi2MoO6 materials was investigated sufficiently by the photodegradation of four different kinds of pollutants, such as ciprofloxacin (CIP), bisphenol A (BPA), tetracycline hydrochloride (TC), and methylene blue (MB). The improved photocatalytic activity was observed for CQD modified Bi2MoO6 samples compared with pure Bi2MoO6 under visible light irradiation. The CQD modified Bi2MoO6 photocatalysts with a CQD content of 2 wt% exhibited the optimum photocatalytic activity, which was found to increase by about 5 times than that of the pure Bi2MoO6 for the photodegradation of CIP. This improvement was attributed to the crucial role of CQDs, which acted as a photocenter for absorbing solar light, a charge separation center for suppressing charge recombination, and a catalytic center for pollutant photo-degradation. The main active species were determined to be ˙OH and O˙-(2) by the ESR technique and analyzed by calculations as well as XPS valence spectra, and a possible photocatalytic mechanism was also proposed.


A polysomnographic and clinical study of sleep disorders in patients with Behçet and neuro-Behçet syndrome.
Brainstem is the most common site of involvement in neuro-Behçet syndrome (NBS). On the other hand, the critical importance of this anatomical region in the regulation of sleep has been disregarded in the literature. We aimed to investigate the microstructure of sleep in patients with Behçet syndrome (BS) and NBS. Patients were allocated to 2 groups: (1) BS without any neurological involvement and (2) NBS with brainstem lesions only. A control group was also enrolled in this study. The comparison of polysomnographic parameters between all patients (BS and NBS) with the control group showed that sleep onset was longer (p = 0.006), the duration of superficial NREM sleep stage (N2) was significantly longer (p = 0.018), and the respiratory disturbance index was significantly higher (p = 0.034) in patients. Sleep apnea and restless legs syndrome are more commonly observed in BS and NBS. Our findings emphasize the importance of questioning the quality of sleep and its disorders in patients with BS in order to better handle the common somatic complaints in these patients, such as fatigue or daytime sleepiness


Microwave-assisted extraction and rapid isolation of ursolic acid from the leaves of Eucalyptus × hybrida Maiden and its quantification using HPLC-diode array technique.
Ursolic acid (UA) is the most important bioactive phytoconstituent of Eucalyptus × hybrida Maiden leaves and exhibits anticancer, antimutagenic, anti-inflammatory, antioxidative, and antiprotozoal activities. In this study, microwave-assisted extraction technique was employed for rapid isolation of UA from the leaves of Eucalyptus × hybrida and simultaneously HPLC-diode array method was developed for the quantification of UA. Effects of several experimental parameters on the extraction efficiencies of UA, such as type and volume of extraction solvents, microwave power and extraction time, were evaluated. The optimal extraction conditions were found to be 20 mL of a mixture of chloroform/methanol, 60:40; liquid-to-material ratio, 4:1; preleaching time, 10 min; microwave power, 600 W; temperature, 50°C; and microwave irradiation time, 5 min. Under the optimum conditions, the yield of UA was found to be 1.95 ± 0.08% in the dry leaves of Eucalyptus × hybrida. The results showed that microwave-assisted extraction is a more rapid extraction method with higher yield and lower solvent consumptions than the conventional method. It is a faster, convenient, and appropriate method and it may be used for rapid isolation and quantification of UA and other important phytoconstituents present in the leaves of Eucalyptus × hybrida.


Intraperiplasmic growth of Bdellovibrio bacteriovorus 109J: N-deacetylation of Escherichia coli peptidoglycan amino sugars.
During intraperiplasmic growth of Bdellovibrio bacteriovorus on Escherichia coli, the substrate cell peptidoglycan is extensively modified as it is converted to bdelloplast peptidoglycan. The initially lysozyme-sensitive peptidoglycan of E. coli was rapidly converted to a lysozyme-resistant form. The conversion was due to the N-deacetylation of a large portion of the peptidoglycan amino sugars. Chemically acetylating the isolated peptidoglycan restored its sensitivity to lysozyme digestion. However, approximately half of the products of lysozyme digestion exhibited hydrophobic interactions that were shown not to be due to the presence of protein. This suggests that a molecule capable of hydrophobic interactions, other than protein, becomes linked to the bdelloplast peptidoglycan. The data also suggest that much of the Braun lipoprotein is removed from the E. coli peptidoglycan early during bdellovibrio development.


Whole sediment toxicity identification evaluation tools for pyrethroid insecticides: III. Temperature manipulation.
Since the toxicity of pyrethroid insecticides is known to increase at low temperatures, the use of temperature manipulation was explored as a whole-sediment toxicity identification evaluation (TIE) tool to help identify sediment samples in which pyrethroid insecticides are responsible for observed toxicity. The amphipod Hyalella azteca is commonly used for toxicity testing of sediments at a 23 degrees C test temperature. However, a temperature reduction to 18 degrees C doubled the toxicity of pyrethroids, and a further reduction to 13 degrees C tripled their toxicity. A similar response, though less dramatic, was found for 1,1-bis(p-chlorophenyl)-2,2,2-trichloroethane (DDT), and dissimilar temperature responses were seen for cadmium and the insecticide chlorpyrifos. Tests with field-collected sediments containing pyrethroids and/or chlorpyrifos showed the expected thermal dependency in nearly all instances. The inverse relationship between temperature and toxicity provides a simple approach to help establish when pyrethroids are the principal toxicant in a sediment sample that could be used as a supplemental tool in concert with chemical analysis or other TIE manipulations. The phenomenon appears to be, in part, a consequence of a reduced ability to biotransform the toxic parent compound at cooler temperatures. The strong dependence of pyrethroid toxicity on temperature has important ramifications for predicting their environmental effects, and the standard test temperature of 23 degrees C dramatically underestimates risk to resident fauna during the cooler months.


The development of a quality assessment tool for ambulance patient care records.
A retrospective cohort study of the 2002 Victorian prehospital emergency care documentation completed by ambulance paramedics had the objectives: (i) to design and implement a quality assessment tool to determine the quality of the ambulance patient care record (PCR)information; and (ii) to identify critical demographic and clinical items on the ambulance PCR that needed improvement. The study outcomes included a functioning quality assessment tool and associated user guide for prehospital use, and the identification of three critical PCR components requiring improvement. Ninety percent of PCRs passed the quality assessment; 10% (approximately 5 300) contained measurably poor or incomplete documentation.


Assessment of radiolucent lines around the Oxford unicompartmental knee replacement: sensitivity and specificity for loosening.
The Oxford unicompartmental knee replacement gives good results in patients with symptomatic osteoarthritis of the medial compartment. Previous studies have suggested that the presence of radiolucent lines (RLLs) does not reflect a poor outcome in such patients. However, the reliability and validity of this assessment have not been determined. Our aim was to assess the intra- and interobserver reliability and the sensitivity and specificity of the assessment of RLLs around both tibial and femoral components using standard radiographs. Two reviewers assessed the radiographs of 45 patients who had loosening of the tibial or femoral component confirmed at revision surgery and compared them with those of a series of 45 asymptomatic patients matched for age and gender. The results suggested that, using standard radiographs, tibial RLLs were 63.6% sensitive and 94.4% specific and femoral RLLs 63.9% sensitive and 72.7% specific for loosening. Overall intra- and interobserver reliability was highly variable, but zonal analysis showed that lucency at the tip of the femoral peg was significantly associated with loosening of the femoral component. Fluoroscopically guided radiographs may improve the zonal reliability of the assessment of RLLs, but further independent and comparative studies are required. In the meantime, the innocence of the physiological RLLs detected by standard radiographs should be viewed with caution.


Assessing psychological well-being: self-report instruments for the NIH Toolbox.
Psychological well-being (PWB) has a significant relationship with physical and mental health. As a part of the NIH Toolbox for the Assessment of Neurological and Behavioral Function, we developed self-report item banks and short forms to assess PWB. Expert feedback and literature review informed the selection of PWB concepts and the development of item pools for positive affect, life satisfaction, and meaning and purpose. Items were tested with a community-dwelling US Internet panel sample of adults aged 18 and above (N = 552). Classical and item response theory (IRT) approaches were used to evaluate unidimensionality, fit of items to the overall measure, and calibrations of those items, including differential item function (DIF). IRT-calibrated item banks were produced for positive affect (34 items), life satisfaction (16 items), and meaning and purpose (18 items). Their psychometric properties were supported based on the results of factor analysis, fit statistics, and DIF evaluation. All banks measured the concepts precisely (reliability ≥0.90) for more than 98% of participants. These adult scales and item banks for PWB provide the flexibility, efficiency, and precision necessary to promote future epidemiological, observational, and intervention research on the relationship of PWB with physical and mental health.


Neoadjuvant chemotherapy in malignant fibrous histiocytoma of bone and in osteosarcoma located in the extremities: analogies and differences between the two tumors.
Malignant fibrous histiocytoma (MFH) is a rare bone tumor usually treated like osteosarcoma. Studies on analogies and differences between the two tumors have seldom been reported. Between March 1982 and December 1994, 51 patients with high-grade MFH of bone and 390 with high-grade osteosarcoma were treated with the same regimen of neoadjuvant chemotherapy. All of the tumors in both groups were located in the limbs. Preoperative chemotherapy was performed according to three different, successively activated, regimens consisting of MTX/CDP intraarterially, MTX/CDP/ADM, and MTX/CDP/ADM//IFO. The rate of limb salvage was the same in both the MFH (92%) and osteosarcoma (85%) patients. MFH showed a statistically significantly lower rate of good histologic response, 90% or more tumor necrosis (27% vs. 67%, P = 0.00001) for all three regimens. Despite this low chemosensitivity, the disease-free survivals of the two neoplasms were similar (67% vs. 65%). In terms of histologic response to primary chemotherapy, MFH has a lower chemosensitivity than osteosarcoma. Nevertheless, the two tumors have similar prognoses when treated with chemotherapy regimens based on MTX, CDP, ADM and IFO.


A soluble form of the receptor for advanced glycation endproducts (RAGE) is produced by proteolytic cleavage of the membrane-bound form by the sheddase a disintegrin and metalloprotease 10 (ADAM10).
The receptor for advanced glycation endproducts (RAGE) mediates responses to cell danger and stress. When bound by its many ligands (which include advanced glycation endproducts, certain members of the S100/calgranulin family, extracellular high-mobility group box 1, the integrin Mac-1, amyloid beta-peptide and fibrils), RAGE activates programs responsible for acute and chronic inflammation. RAGE is therefore also involved in cancer progression, diabetes, atherosclerosis, and Alzheimer's disease. RAGE has several isoforms deriving from alternative splicing, including a soluble form called endogenous secretory RAGE (esRAGE). We show here that most soluble RAGE, either produced by cell lines or present in human blood, is not recognized by an anti-esRAGE antibody. Cells transfected with the cDNA for full-length RAGE, and thus not expressing esRAGE, produce a form of soluble RAGE, cleaved RAGE (cRAGE) that derives from proteolytic cleavage of the membrane-bound molecules and acts as a decoy receptor. By screening chemical inhibitors and genetically modified mouse embryonic fibroblasts (MEFs), we identify the sheddase ADAM10 as a membrane protease responsible for RAGE cleavage. Binding of its ligand HMGB1 promotes RAGE shedding. Our data do not disprove the interpretation that high levels of soluble forms of RAGE protect against chronic inflammation, but rather suggest that they correlate with high levels of ongoing inflammation.


Selectivity of lexical-semantic disorders in Polish-speaking patients with aphasia: evidence from single-word comprehension.
Several neuropsychological studies have shown that patients with brain damage may demonstrate selective category-specific deficits of auditory comprehension. The present paper reports on an investigation of aphasic patients' preserved ability to perform a semantic task on spoken words despite severe impairment in auditory comprehension, as shown by failure in matching spoken words to pictured objects. Twenty-six aphasic patients (11 women and 15 men) with impaired speech comprehension due to a left-hemisphere ischaemic stroke were examined; all were right-handed and native speakers of Polish. Six narrowly defined semantic categories for which dissociations have been reported are colors, body parts, animals, food, objects (mostly tools), and means of transportation. An analysis using one-way ANOVA with repeated measures in conjunction with the Lambda-Wilks Test revealed significant discrepancies among these categories in aphasic patients, who had much more difficulty comprehending names of colors than they did comprehending names of other objects (F((5,21))=13.15; p<.001). Animals were most often the easiest category to understand. The possibility of a simple explanation in terms of word frequency and/or visual complexity was ruled out. Evidence from the present study support the position that so called "global" aphasia is an imprecise term and should be redefined. These results are discussed within the connectionist and modular perspectives on category-specific deficits in aphasia.


Anterior vertebral body screw position placed thoracoscopically: a function of anatomy and surgeon experience in a porcine model.
Manual digital measurements, in a porcine model of thoracoscopic anterior instrumentation, divided into two studies: 1) comparing the actual screw position within each vertebra with the perceived ideal screw position and correlation with the learning curve of the surgeon; and 2) a quantitative analysis of the vertebral bodies from T3 to T12 to determine the ideal screw insertion point and trajectory. To investigate whether the position of thoracoscopically placed screws is related to the portion of the thoracic spine instrumented and to the experience of the surgeon and to define the ideal starting position and direction of anterior thoracic screws in a porcine model. Anterior screw insertion using a thoracoscopic approach is generally considered to be technically demanding and potentially dangerous. To our knowledge, there is no study analyzing the proper position of anterior vertebral screws using thoracoscopic technique and no study analyzing the ideal starting position of anterior vertebral body screws. In Study 1, 26 pigs were assigned to two groups (early experience, n = 16; late experience, n = 10) and underwent thoracoscopic anterior instrumentation and fusion from T5 to T10. The screw position was determined in the sagittal plane and in the frontal plane, respectively. In Study 2, 10 thoracic pig spines (T3-T12) were obtained. Each vertebra was digitally measured in the midtransverse plane. Using a point directly adjacent to the rib head as the assumed ideal screw starting point, the percent of vertebral body "obscured" by the rib head and the maximum safe anterior and posterior screw insertion angles were determined. In Study 1, screw positions of Group 1 were significantly more anterior from T5 to T7 compared with the ideal screw position (P < 0.05). Most screws were in the ideal position in Group 2. In both groups, the screw position within T5 was too inferior (P < 0.05). The majority of posterior cortical disruptions occurred in the distal vertebrae. Most anterior disruptions occurred in the proximal vertebrae. In Study 2, the percent of vertebral body behind the rib head significantly decreased from T3 (61.5%) to T12 (25.7%) (P < 0.0001). The maximum anterior insertion angle significantly increased from T3 (11 degrees) to T12 (24 degrees) (P < 0.0001) while the maximum posterior insertion angle significantly decreased from T3 (28 degrees) to T12 (11 degrees) (P < 0.0001). The position of thoracoscopically placed vertebral screws is dependent on the level of the spine instrumented and surgeon experience. Screws placed in the proximal thoracic spine tend to be too anterior and too inferior while posterior placement of screws occurs in the distal thoracic spine. When using the rib head as the starting point for anterior screw insertion, a slight anterior angle is required in the distal thoracic spine while a slight posterior insertion angle is required in the proximal thoracic spine. A learning curve does exist, however; the proximal screw positioning continues to be a challenge.


Two cases of severe bronchopneumonia due to influenza A (H3N2) virus: detection of influenza virus gene using reverse transcription polymerase chain reaction.
We report two cases of severe bronchopneumonia due to influenza A (H3N2) virus. The severity of the disease necessitated initiation of empiric therapy based on the present illness and clinical data on admission. Both patients were improved by artificial ventilation with positive end-expiratory pressures and administration of broad spectrum antibiotics and corticosteroids before confirming the diagnosis of viral bronchopneumonia using viral culture and serological tests. Within 24 hours, influenza A (H3N2) virus was identified by amplification of the pathogen genes by reverse transcription polymerase chain reaction (RT-PCR) using the stored bronchoalveolar lavage (BAL) fluids of both cases. This suggests that a combination of detection methods of pathogens using RT-PCR and BAL fluid will facilitate determination of rational treatment aimed at influenza A virus.


Urinary nerve growth factor levels could be a biomarker for overactive bladder symptom: a meta-analysis.
To examine whether urinary tract nerve growth factor (uNGF) could be a biomarker for overactive bladder (OAB) symptom, we conducted a comprehensive meta-analysis of 8 case-control studies. In all the studies considered, patients with OAB symptom had a higher uNGF level compared to healthy people. In addition, patients had a significantly lower uNGF level after successful treatment. In the subgroup analysis, we found that patients with OAB-wet symptom had a higher uNGF level than patients with OAB-dry symptom. However, no significant difference was found between patients with OAB symptom and patients with interstitial cystitis/painful bladder syndrome (IC/PBS) symptom in uNGF/Cr levels. In conclusion, uNGF level could be a useful biomarker for the diagnosis of OAB, a possible biomarker for differentiation between OAB subtypes (wet or dry), and a predictive biomarker for a specific treatment, but it cannot be used as the urinary biomarker for the differential diagnosis of IC/PBS and OAB.


Expression of the c-src protooncogene in human skin tumors.
Retroviral oncogenes are genetic elements, the expression of which is responsible for the transformed phenotype of cells. These genes are derived from normal cellular DNA sequences called cellular protooncogenes, which are present in all human cells and seem to have potential transforming ability in tumors of nonviral origin, since it is possible that they undergo structural alterations and/or changes in their expression. Human skin tumors were analyzed in this study with respect to the expression of the c-src protooncogene, the cellular homologue of the Rous sarcoma virus transforming gene, by measuring the enzymatic activity of its gene product, the pp60c-src kinase activity. Tyrosine-specific kinase activity was detected in all skin tumors tested. The expression pattern of the c-src gene product in the melanomas tested was differential and varying kinase levels in different metastases from the same patient were detected. The elevation of kinase activity as compared to normal skin ranged from about 4- to 20-fold.


[The characteristics of right ventricular function in patients with essential hypertension].
Two-dimensional and pulsed Doppler echocardiography were used to evaluate the function of right ventricule in 166 patients with essential hypertension and the results were compared with those of 79 normal subjects. The pulmonary artery peak flow velocity in 100 patients without left ventricular hypertrophy (LVH) had no significant difference with that of normal controls (0.65 +/- 0.17 vs 0.63 +/- 0.18, P greater than 0.05). The tricuspid early peak flow velocity (E) decreased (0.43 +/- 0.08 vs 0.46 +/- 0.12, P less than 0.01) but the late peak flow velocity (A) increased (0.30 +/- 0.09 vs 0.24 +/- 0.06, P less than 0.001). E/A reduced also (1.57 +/- 0.53 vs 2.02 +/- 0.78, P less than 0.01). The pulmonary peak flow velocity in 66 patients with LVH elevated markedly as compared with those without LVH (0.71 +/- 0.18 vs 0.65 +/- 0.17, P less than 0.001), A increased (0.34 +/- 00.10 vs 0.30 +/- 0.09, P less than 0.001) and E/A decreased (1.29 +/- 0.37 vs 1.57 +/- 0.53, P less than 0.001). The decrement of E/A correlated with the thickness of interventricular septum and the left ventricular mass (r = -0.68 and -0.60 respectively, P less than 0.01). The results show that the diastolic function of right ventricule was damaged in the early stage of essential hypertension but the systolic function remains unchanged. The diastolic function decreased and the systolic function increased further concomitantly with the development of LVH.


Doubling the dose of budesonide versus maintenance treatment in asthma exacerbations.
Previous guidelines recommend doubling the daily dose of maintenance inhaled corticosteroid to treat or prevent progression of exacerbations of asthma. Over a 6 month period a cohort of patients were evaluated prospectively and randomised in a double blind controlled trial to treatment with either a continued maintenance dose (MD) of inhaled corticosteroid or doubling the dose (DD) at the time of an exacerbation. A total of 290 patients were randomised (33% male) and 98 (DD, n = 46) experienced evaluable asthma exacerbations during the study period. Mean (SD) baseline characteristics at randomisation (age 33.5 (14.0) years; forced expiratory volume in 1 second (FEV(1)) 2.8 (0.7) l; peak expiratory flow (PEF) 422.9 (110.5) l/min) were similar in both groups. In the DD group 41% of patients were considered treatment failures because they either required systemic steroids (n = 12), had an unscheduled visit to a physician (n = 1), or their asthma did not return to baseline (n = 6). This did not differ from the MD group in which 40% were treatment failures (n = 9, 0, and 12, respectively; p = 0.94). In patients who regularly take an inhaled corticosteroid, doubling the maintenance dose may not affect the pattern of the exacerbation.


[Long-term results following cholesteatoma surgery].
A long-term review of 140 ears in 133 patients was carried out. All ears had first been operated on in 1971 and 1972 for cholesteatoma in the E.N.T. clinic. A closed technique was used in 110 ears and in the remaining 30 ears an open technique, depending on the pneumatisation of the mastoid, the extent of the cholesteatoma, tubal function and the age and social circumstances of the patient. At that time, second-look operations were carried out on relatively few patients after a closed operation. While the short-term results were favourable, recurrent cholesteatomas still developed 8 to 10 years later; 9 were discovered at this review alone. The long-term incidence of recurrence was 25%. Recurrent cholesteatomas were found in 57% of patients who were first operated on for a cholesteatoma before the age of 21, 24% of those between 21 and 40, and 11% of those between 41 and 60. These results reaffirm the necessity for a planned second-look operation in all young patients and also the importance of outpatient follow-up over many years.


Evaluation of shelter dog activity levels before and during COVID-19 using automated analysis.
Animal shelters have been found to represent stressful environments for pet dogs, both affecting behavior and influencing welfare. The current COVID-19 pandemic has brought to light new uncertainties in animal sheltering practices which may affect shelter dog behavior in unexpected ways. To evaluate this, we analyzed changes in dog activity levels before COVID-19 and during COVID-19 using an automated video analysis within a large, open-admission animal shelter in New York City, USA. Shelter dog activity was analyzed during two two-week long time periods: (i) just before COVID-19 safety measures were put in place (Feb 26-Mar 17, 2020) and (ii) during the COVID-19 quarantine (July 10-23, 2020). During these two periods, video clips of 15.3 second, on average, were taken of participating kennels every hour from approximately 8 am to 8 pm. Using a two-step filtering approach, a matched sample (based on the number of days of observation) of 34 dogs was defined, consisting of 17 dogs in each group (N1/N2 = 17). An automated video analysis of active/non-active behaviors was conducted and compared to manual coding of activity. The automated analysis validated by comparison to manual coding reaching above 79% accuracy. Significant differences in the patterns of shelter dog activity were observed: less activity was observed in the afternoons before COVID-19 restrictions, while during COVID-19, activity remained at a constant average. Together, these findings suggest that 1) COVID-19 lockdown altered shelter dog in-kennel activity, likely due to changes in the shelter environment and 2) automated analysis can be used as a hands-off tool to monitor activity. While this method of analysis presents immense opportunity for future research, we discuss the limitations of automated analysis and guidelines in the context of shelter dogs that can increase accuracy of detection, as well as reflect on policy changes that might be helpful in mediating canine stress in changing shelter environments.


The neural basis of individual differences in mate poaching.
This study tested the hypothesis that individual differences in the activity of the orbitofrontal cortex, a region implicated in value-based decision-making, are associated with the preference for a person with a partner, which could lead to mate poaching. During functional magnetic resonance imaging (fMRI), male participants were presented with facial photographs of (a) attractive females with a partner, (b) attractive females without a partner, (c) unattractive females with a partner, and (d) unattractive females without a partner. The participants were asked to rate the degree to which they desired a romantic relationship with each female using an 8-point scale. The participants rated attractive females higher than unattractive females, and this effect was associated with ventral striatum activation. The participants also indicated lower ratings for females with a partner than for females without a partner, and this effect was associated with parietal cortex activation. As predicted, the participants characterized by higher orbitofrontal activity demonstrated a greater willingness to engage in a romantic relationship with females who have a partner compared with females who do not have a partner. These results are the first to provide a possible neural explanation for why certain individuals are willing to engage in mate poaching.


Gender Bias in Letters of Recommendation: Relevance to Urology Match Outcomes and Pursuit of Fellowship Training/Academic Career.
To review applications to a single urology residency program to determine application characteristics predictive of (1) successful match into urology residency and (2) pursuit of fellowship training and/or academic practice after completion of residency. Our principal variables of interest were gender bias as assessed in letters of recommendation (LOR), personal statements, Medical Student Performance Evaluation (MSPE), race, and gender. Applications submitted to our urology residency program in the 2014 cycle were reviewed. Twenty-three variables were analyzed, including applicant demographics, application materials, and gender bias. Deidentified text from LOR, personal statements, and MSPE was evaluated for gender bias using an open-source gender bias calculator. A subanalysis of applicants who matched at a top 25 urology program was performed. Logistic regression analysis was performed to identify applicant variables associated with (1) match success and (2) fellowship training or academic employment as of September 2021. Two hundred and twenty-two completed applications were analyzed. First authorship of a published manuscript was significantly associated with greater odds of matching. Female gender and top 25 medical school attendance were both significant predictors of matching at a top 25 urology program. The number of first-author publications was associated with completion of fellowship training or current employment in an academic position. First-author publications are the most important preinterview determinant of match success and subsequent pursuit of academic practice/fellowship training. Certain applicant characteristics are associated with matching at highly ranked programs. Gender bias in application materials (including LOR) does not appear to exert a significant influence on match and early career outcomes.


Assessing body composition with DXA and bioimpedance: effects of obesity, physical activity, and age.
This study evaluated to what extent dual-energy X-ray absorptiometry (DXA) and two types of bioimpedance analysis (BIA) yield similar results for body fat mass (FM) in men and women with different levels of obesity and physical activity (PA). The study population consisted of 37-81-year-old Finnish people (82 men and 86 women). FM% was estimated using DXA (GE Lunar Prodigy) and two BIA devices (InBody (720) and Tanita BC 418 MA). Subjects were divided into normal, overweight, and obese groups on the basis of clinical cutoff points of BMI, and into low PA (LPA) and high PA (HPA) groups. Agreement between the devices was calculated by using the Bland-Altman analysis. Compared to DXA, both BIA devices provided on average 2-6% lower values for FM% in normal BMI men, in women in all BMI categories, and in both genders in both HPA and LPA groups. In obese men, the differences were smaller. The two BIA devices provided similar means for groups. Differences between the two BIA devices with increasing FM% were a result of the InBody (720) not including age in their algorithm for estimating body composition. BIA methods provided systematically lower values for FM than DXA. However, the differences depend on gender and body weight status pointing out the importance of considering these when identifying people with excess FM.


Synchronous tropical South China Sea SST change and Greenland warming during deglaciation.
The tropical ocean plays a major role in global climate. It is therefore crucial to establish the precise phase between tropical and high-latitude climate variability during past abrupt climate events in order to gain insight into the mechanisms of global climate change. Here we present alkenone sea surface temperature (SST) records from the tropical South China Sea that show an abrupt temperature increase of at least 1 degrees C at the end of the last glacial period. Within the recognized dating uncertainties, this SST increase is synchronous with the Bølling warming observed at 14.6 thousand years ago in the Greenland Ice Sheet Project 2 ice core.


Differential study of two strains of Toxoplasma gondii.
This study aimed to differentiate between the virulent and avirulent strains of T. gondii by studying morphometric measurement using C.I.A.S., determination of the relative DNA content and extract, SDS poly acrylamide gel electrophoresis and isoenzyme on cellulose acetate gel. Identifying these differences may be useful in studying different disease types and in pathogenic mechanisms. Besides, the presence of common proteins between them may be of value for diagnostic purposes and for formulation of vaccines.


Thyroid function and polyamines. II. Thyrotropin stimulation of polyamine biosynthesis in the rat thyroid.
Thyrotropin (TSH) injected intraperitoneally caused a significant rise in the activity of L-ornithine decarboxylase of rat thyroid within 4 hr. TSH-releasing hormone (TRH) also increased the activity of this enzyme in a dose-related manner and of putrescine-activated S-adenosyl-L-methionine decarboxylase, probably through the increased secretion of TSH from the pituitary. Administration of methylthiouracil (MTU) in the drinking water to rats resulted in an increase of these two enzymes in the thyroid. Maximal activation was observed 4 days after the initiation of MTU treatment for both enzymes. The intracellular level of putrescine and spermidine increased during the rapid phase of thyroid enlargement caused by MTU treatment. The spermine concentration, however, was relatively constant during the treatment. The RNA/DNA ratio followed a pattern very similar to that exhibited by the spermidine/spermine ratio. These results indicate that TSH stimulates the biosynthesis of polyamines by increasing the activity of the decarboxylases in the thyroid. The findings further suggest that polyamines play a role in the regulation of thyroid growth and nucleic acid metabolism.


[Bone tumors. Their prevalence].
1,200 cases of bone tumors were reviewed from the Orthopedic Hospital "Magdalena de las Salinas" between 1982 and 1989. 66.7% of them were benign tumors, 14.3% pseudotumoral lesions, 10.1% metastatic tumors, 8.6% primary malignant tumors, and 0.3% were malignant invasive tumors to bone. The most frequently found benign tumors were: osteochondroma, enchondroma and giant cell tumor; the most frequent pseudotumoral lesions were metaphyseal fibrous defects, solitary bone cysts and fibrous dysplasia; the most frequent primary malignant tumor was osteosarcoma. Prevalence, frequency, distribution, sex, most frequent ages, affected bones, multicentricity, and aggregated fractures, as well as the site of the primary lesion in metastatic and invading tumors, were similar to those reported in the classical series.


[Micro-neurosurgical treatment of cranionasal tumors in combination with transnasal endoscopy].
To explore an effective microsurgical approach to the treatment of cranionasal tumors. A retrospective review of 18 micro-neurosurgical patients with cranionasal tumors (June 2005 to June 2007) was undertaken. All of the 18 patients were treated with subfrontal approaches in combination with transnasal endoscopy. Tumors were resected in the stage-one operations (14 were totally resected and 4 were subtotally resected). The anterior skull bases were reconstructed. Transient CSF rhinorrhea was found in two cases. All of the patients experienced good recoveries, with no operative death. The follow up after 5 to 29 months revealed that only four patients had tumor recurrence. Three patients lost in the follow up. Subfrontal microsurgical operation combined with transnasal endoscopy is an effective approach to the treatment of cranionasal tumors. It enables high total resection rate and has low complications.


Glial cell line-derived neurotrophic factor promotes sleep in rats and rabbits.
Various growth factors (e.g., growth hormone-releasing hormone, acidic fibroblast growth factor, nerve growth factor, brain-derived neurotrophic factor, and interleukin-1) are implicated in sleep regulation. It is hypothesized that neuronal activity enhances the production of such growth factors, and they in turn form part of the sleep regulatory mechanism. Glial cell line-derived neurotrophic factor (GDNF) promotes development, differentiation, maintenance, and regeneration of neurons, and its production is induced by well-characterized sleep regulatory substances such as interleukin-1 and tumor necrosis factor. Therefore, we investigated whether GDNF would promote sleep. Twenty-six male Sprague-Dawley rats and 30 male New Zealand White rabbits were surgically implanted with electroencephalogram (EEG) and electromyogram (EMG; rats only) electrodes, a brain thermistor, and a lateral intracerebroventricular cannula. The animals were injected intracerebroventricularly with pyrogen-free saline and on a separate day with one of the following doses of GDNF: 5, 50, and 500 ng in rabbits and 50 and 500 ng in rats. The EEG, brain temperature, EMG (in rats), and motor activity (in rabbits) were recorded for 23 h after the intracerebroventricular injection. GDNF (500-ng dose) increased the time spent in nonrapid eye movement sleep in both rats and rabbits. Rapid eye movement sleep was not affected by the lower doses of GDNF but was inhibited in rabbits after the high dose. EEG slow-wave activity was not affected by GDNF. The current results provide further evidence that various growth factors are involved in sleep regulation.


The uptake and efflux of L-phenylalanine and L-tyrosine from rat brain cerebral cortex slices.
The uptake of radioactive l-phenylalanine and l-tyrosine into the tissue water of rat brain cerebral cortex slices was shown to have saturation kinetics. The apparent K(m) for the uptake of l-phenylalanine was 0.86mm and for l-tyrosine was 1.64mm; for phenylalanine the apparent V(max.) value was 0.64mumol/min per ml of tissue water, and for l-tyrosine it was 0.98mumol/min per ml of tissue water. The accumulation of the two amino acids by the tissue was depressed in the absence of O(2), at 0 degrees C, or in the presence of metabolic inhibitors. The influxes and effluxes of both l-isomers were more rapid than those of the d-isomers. Competition between these two amino acids and each with l-tryptophan in respect of uptake into tissue water was shown. Their rates of influx were faster, and rates of efflux were slower, in the presence than in the absence of Na(+). It was concluded that these amino acids were taken up by active transport via a carrier mechanism.


Women and hospice care.
The hospice movement has grown extensively in the last ten years. This article examines the role of women in hospice, components of hospice care and a profile of 544 admissions to an inpatient hospice unit over a five year period. Case studies are given to illustrate the needs of women in hospice. Regulatory processes facing hospice programs today are reviewed.


Digit-sucking: a review of the literature, clinical observations and treatment recommendations.
Prolonged digit-sucking habits can have a negative impact on dental, speech, physical and emotional development. Children can be helped to eliminate their sucking habits quickly, without coercion, and in a positive way. Admittedly, most of my data is based purely on clinical observation. However, clinical observation is the basis for classical scientific investigation. I welcome this investigation; it is long overdue. Much of the literature available to parents appears to take a rather cavalier attitude with regard to the dental consequences of sucking habits. Parents are frequently left with the impression that no harm is done to the teeth. This perspective is based on the assumption that most children stop the sucking behavior before permanent teeth erupt into the mouth. Therefore, any dental malocclusion in the primary teeth will often be transitional or will spontaneously self-correct. However, as demonstrated by Kelly, et al (1973), millions of children do not discontinue their sucking habits before permanent teeth erupt. In addition, stress levels in today's world are probably significantly greater than when this study was done. Because stress is a powerful stimulus for the sucking activity, it is highly probable that if a similar study were done today, it would produce a substantially greater number of individuals with prolonged sucking habits. The active pursuit of insurance benefits for the Certified Orofacial Myologist providing therapy for the elimination of these habits will be germane to prevention. Research is needed to accomplish this goal including: investigation of the incidence of prolonged digit-sucking habits; the impact of these habits on the entire stomatognathic system, speech development and emotional health; efficacy of treatment (including psychological response) both through the utilization of habit appliances and motivational therapy. The Certified Orofacial Myologist provides many valuable therapeutic programs to enhance dental and speech development. In the words of our president, Marge Snow at the 25th Anniversary of the founding of The International Association of Orofacial Myology, "we are morally and ethically challenged with making this treatment available to everyone". Roberta Pierce (1988) in her paper, "Treatment For The Young Child" wrote, "The overall goal is to create as normal an environment as possible so as to facilitate normal growth and development." The early elimination of digit-sucking habits is one of the most important therapeutic services offered by the Certified Orofacial Myologist because it provides the most benefit toward prevention and the well-being of the whole person. Treatment after the pathology is fully developed is invasive, not cost-effective and does not provide as good a clinical result. Little mouths deserve a big chance to grow healthy and straight.


Sporosalibacterium faouarense gen. nov., sp. nov., a moderately halophilic bacterium isolated from oil-contaminated soil.
A novel strictly anaerobic, moderately halophilic and mesophilic bacterium, designated strain SOL3f37(T), was isolated from a hydrocarbon-polluted soil surrounding a deep petroleum environment located in south Tunisia. Cells of strain SOL3f37(T) stained Gram-positive and were motile, straight and spore-forming. Strain SOL3f37(T) had a typical Gram-positive-type cell-wall structure, unlike the thick, multilayered cell wall of its closest relative Clostridiisalibacter paucivorans. The major fatty acids were iso-C(15 : 0) (41 %), iso-C(14 : 0) 3-OH and/or iso-C(15 : 0) dimethyl acetal (21.6 %), iso-C(13 : 0) (4.4 %), anteiso-C(15 : 0) (3.9 %) and iso-C(15 : 1) (2.8 %). Strain SOL3f37(T) grew between 20 and 48 °C (optimum 40 °C) and at pH 6.2-8.1 (optimum pH 6.9). Strain SOL3f37(T) required at least 0.5 NaCl l(-1) and grew in the presence of NaCl concentrations up to 150 g l(-1) (optimum 40 g l(-1)). Yeast extract (2 g l(-1)) was required for degradation of pyruvate, fumarate, fructose, glucose and mannitol. Also, strain SOL3f37(T) grew heterotrophically on yeast extract, peptone and bio-Trypticase, but was unable to grow on Casamino acids. Sulfate, thiosulfate, sulfite, elemental sulfur, fumarate, nitrate and nitrite were not reduced. The DNA G+C content was 30.7 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain SOL3f37(T) was a member of the family Clostridiaceae in the order Clostridiales; strain SOL3f37(T) was related to members of various genera of the family Clostridiaceae. It exhibited highest 16S rRNA gene sequence similarity (93.4 %) with Clostridiisalibacter paucivorans 37HS60(T), 91.8 % with Thermohalobacter berrensis CTT3(T) and 91.7 % with Caloranaerobacter azorensis MV1087(T). On the basis of genotypic, phenotypic and phylogenetic data, it is suggested that strain SOL3f37(T) represents a novel species in a new genus. The name Sporosalibacterium faouarense gen. nov., sp. nov. is proposed, with SOL3f37(T) (=DSM 21485(T) =JCM 15487(T)) as the type strain of Sporosalibacterium faouarense.


[Combined study of leukemic blasts using DNA autoradiography and cytophotometry].
The 3H-thymidine incorporation capacity and DNA contents were studied in the same bone marrow blasts in 4 patients suffering from acute leucaemia. The blast populations were both proliferating and non-proliferating. Actively proliferating cells were blasts of large and medium diameters. Mitotically inactive cells were blasts of medium and small diameters. Cells that stopped dividing were established as being in period G1 of the cell cycle. A high variation in DNA contents was noted in blasts unlabeled with 3H-thymidine, a possible reason of the above phenomenon being discussed.


Total and abdominal adiposity are associated with inflammation in older adults using a factor analysis approach.
Obesity-related increases in multiple inflammatory markers may contribute to the persistent subclinical inflammation common with advancing age. However, it is unclear if a specific combination of markers reflects the underlying inflammatory state. We used factor analysis to identify inflammatory factor(s) and examine their associations with adiposity in older adults at risk for disability. Adiponectin, CRP, IL-1ra, IL-1sRII, IL-2sRα, IL-6, IL-6sR, IL-8, IL-15, sTNFRI, sTNFRII, and TNF-α were measured in 179 participants from the Lifestyle Interventions and Independence for Elders Pilot (Mean ± SD age 77 ± 4 years, 76% white, 70% women). Body mass index, waist circumference, and total fat mass were assessed by anthropometry and dual-energy x-ray absorptiometry. IL-2sRα, sTNFRI, and sTNFRII loaded highest on the first factor (factor 1). CRP, IL-1ra, and IL-6 loaded highest on the second factor (factor 2). Factor 2, but not factor 1, was positively associated with 1-SD increments in waist circumference (β = 0.160 ± 0.057, p = .005), body mass index (β = 0.132 ± 0.053, p = .01), and total fat mass (β = 0.126 ± 0.053, p = .02) after adjusting for age, gender, race/ethnicity, site, smoking, anti-inflammatory medications, comorbidity index, health-related quality of life, and physical function. These associations remained significant after further adjustment for grip strength, but only waist circumference remained associated with inflammation after adjusting for total lean mass. There were no significant interactions between adiposity and muscle mass or strength for either factor. Greater total and abdominal adiposity are associated with higher levels of an inflammatory factor related to CRP, IL-1ra, and IL-6 in older adults, which may provide a clinically useful measure of inflammation in this population.