The Putative Role of Viruses, Bacteria, and Chronic Fungal Biotoxin Exposure in the Genesis of Intractable Fatigue Accompanied by Cognitive and Physical Disability.
Patients who present with severe intractable apparently idiopathic fatigue accompanied by profound physical and or cognitive disability present a significant therapeutic challenge. The effect of psychological counseling is limited, with significant but very slight improvements in psychometric measures of fatigue and disability but no improvement on scientific measures of physical impairment compared to controls. Similarly, exercise regimes either produce significant, but practically unimportant, benefit or provoke symptom exacerbation. Many such patients are afforded the exclusionary, non-specific diagnosis of chronic fatigue syndrome if rudimentary testing fails to discover the cause of their symptoms. More sophisticated investigations often reveal the presence of a range of pathogens capable of establishing life-long infections with sophisticated immune evasion strategies, including Parvoviruses, HHV6, variants of Epstein-Barr, Cytomegalovirus, Mycoplasma, and Borrelia burgdorferi. Other patients have a history of chronic fungal or other biotoxin exposure. Herein, we explain the epigenetic factors that may render such individuals susceptible to the chronic pathology induced by such agents, how such agents induce pathology, and, indeed, how such pathology can persist and even amplify even when infections have cleared or when biotoxin exposure has ceased. The presence of active, reactivated, or even latent Herpes virus could be a potential source of intractable fatigue accompanied by profound physical and or cognitive disability in some patients, and the same may be true of persistent Parvovirus B12 and mycoplasma infection. A history of chronic mold exposure is a feasible explanation for such symptoms, as is the presence of B. burgdorferi. The complex tropism, life cycles, genetic variability, and low titer of many of these pathogens makes their detection in blood a challenge. Examination of lymphoid tissue or CSF in such circumstances may be warranted.


Theory of midinfrared absorption microspectroscopy: I. Homogeneous samples.
Midinfrared (IR) microspectroscopy is widely employed for spatially localized spectral analyses. A comprehensive theoretical model for the technique, however, has not been previously proposed. In this paper, rigorous theory is presented for IR absorption microspectroscopy by using Maxwell's equations to model beam propagation. Focusing effects, material dispersion, and the geometry of the sample are accounted to predict spectral response for homogeneous samples. Predictions are validated experimentally using Fourier transform IR (FT-IR) microspectroscopic examination of a photoresist. The results emphasize that meaningful interpretation of IR microspectroscopic data must involve an understanding of the coupled optical effects associated with the sample, substrate properties, and microscopy configuration. Simulations provide guidance for developing experimental methods and future instrument design by quantifying distortions in the recorded data. Distortions are especially severe for transflection mode and for samples mounted on certain substrates. Last, the model generalizes to rigorously consider the effects of focusing. While spectral analyses range from examining gross spectral features to assessing subtle features using advanced chemometrics, the limitations imposed by these effects in the data acquisition on the information available are less clear. The distorting effects are shown to be larger than noise levels seen in modern spectrometers. Hence, the model provides a framework to quantify spectral distortions that may limit the accuracy of information or present confounding effects in microspectroscopy.


The accuracy of transcutaneous PCO2 in subjects with severe brain injury: a comparison with end-tidal PCO2.
In patients suffering from brain injury, end-tidal PCO2 (PETCO2 ) monitoring is controversial, but transcutaneous PCO2 (PtcCO2 ), which is noninvasive and utilizes immediate display, may be an alternative method. We hypothesized that PtcCO2 would be more accurate than PETCO2 for monitoring PaCO2 in patients with severe brain injury. A prospective observational study included consecutive mechanically ventilated adult subjects who had acute brain injury and an arterial catheter in place. When an arterial blood gas analysis was required, the PETCO2 and PtcCO2 values were simultaneously recorded. The agreement between the PETCO2 , PtcCO2 , and PaCO2 measurements (reference) was determined using the Bland-Altman method. The number of outliers defined by the formula ([PETCO2 or PtcCO2 ] - PaCO2 ) > ± 4 mm Hg indicated the proportion of measurements that were considered clinically unacceptable. A total of 25 subjects were included in the study, and 85 simultaneous measurements of PaCO2 , PtcCO2 , and PETCO2 were obtained. The bias and precision between PaCO2 and PtcCO2 were -0.75 and 6.23 mm Hg, respectively. The limits of agreement ranged from -12.97 to 11.47 mm Hg. The bias and precision between PaCO2 and PETCO2 were 0.68 and 5.82 mm Hg, respectively. The limits of agreement ranged from -10.72 to 12.08 mm Hg. There were 34 (40%) outliers for the PtcCO2 sensor and 34 (40%) outliers for the PETCO2 sensor (P > .99). The accuracy of PtcCO2 was not superior to that of PETCO2 for assessing PCO2 levels and should not be used to monitor these levels in subjects with severe brain injury.


Identifying and elucidating the roles of Y198N and Y204F mutations in the PAH enzyme through molecular dynamic simulations.
Phenylketonuria is an autosomal recessive disorder caused by mutations in the phenylalanine hydroxylase gene. In phenylketonuria causes various symptoms including severe mental retardation. PAH gene of a classical Phenylketonuria patient was sequenced, and two novel heterozygous mutations, p.Y198N and p.Y204F, were found. This study aimed to reveal the impacts of these variants on the structural stability of the PAH enzyme. In-silico analyses using prediction tools and molecular dynamics simulations were performed. Mutations were introduced to the wild type catalytic monomer and full length tetramer crystal structures. Variant pathogenicity analyses predicted p.Y198N to be damaging, and p.Y204F to be benign by some prediction tools and damaging by others. Simulations suggested p.Y198N mutation cause significant fluctuations in the spatial organization of two catalytic residues in the temperature accelerated MD simulations with the monomer and increased root-mean-square deviations in the tetramer structure. p.Y204F causes noticeable changes in the spatial positioning of T278 suggesting a possible segregation from the catalytic site in temperature accelerated MD simulations with the monomer. This mutation also leads to increased root-mean-square fluctuations in the regulatory domain which may lead to conformational change resulting in inhibition of dimerization and enzyme activation. Our study reports two novel mutations in the PAH gene and gives insight to their effects on the PAH activity. MD simulations did not yield conclusive results that explains the phenotype but gave plausible insight to possible effects which should be investigated further with in-silico and in-vitro studies to assess the roles of these mutations in etiology of PKU. Communicated by Ramaswamy H. Sarma.


Guideline for the prevention of acute chemotherapy-induced nausea and vomiting in pediatric cancer patients: A focused update.
This update of the 2013 clinical practice guideline provides clinicians with guidance regarding the use of aprepitant and palonosetron for the prevention of acute chemotherapy-induced nausea and vomiting (CINV) in children. The recommendations were based on three systematic reviews. Substantive changes were made to the guideline recommendations including the inclusion of palonosetron to the 5-HT3 antagonists recommended for children receiving highly emetogenic chemotherapy (HEC) and the recommendation of aprepitant for children 6 months of age or older receiving HEC. To optimize CINV control in children, future work must focus on closing critical research gaps.


Making the case for completion bonuses in clinical trials.
Attrition is a serious problem in many clinical trials. The practice of offering completion bonuses-financial incentives offered to participants on the condition that they remain in a trial until they reach a prespecified study endpoint-is one means of addressing attrition. Despite their practical appeal, however, completion bonuses remain ethically controversial due to concern that they will coerce or unduly influence participants to not exercise their right to withdraw from a trial. Although this interaction with the right to withdraw does render completion bonuses conceptually distinct from other incentive payments offered to research participants, we argue here that completion bonuses are never coercive and, in the context of effective institutional review board oversight, are unlikely to be unduly influential. Nonetheless, because completion bonuses may in some cases still encourage unreasonable continued participation in a study, additional safeguards are appropriate. Rejecting completion bonuses entirely is, however, unnecessary and would problematically fail to address the significant ethical problems associated with participant attrition.


Bilingual Conversion in the Translation of ICH Terms: A Study on the Psychological Processes of Translators.
Intangible cultural heritage (ICH) terms are unique expressions of cultural knowledge and practices that are deeply rooted in a particular region or community. Accurately translating ICH terms is crucial for sharing this knowledge and promoting cross-cultural understanding. However, the complex nature of ICH terms, including their regional and cultural specificity, means that mistranslations can easily occur during the translation process. In addition to linguistic and cultural barriers, psychological factors can also impact the accuracy of translations. Translation psychology, a new discipline that examines the psychological process of bilingual conversion and its cognitive mechanisms, can provide insights into the translation of ICH terms. This paper proposes an information processing model of translating ICH terms based on translation psychology, which examines the psychological processes of translators during the translation of Chinese ICH terms into English. Through questionnaires and translation exercises, the study compares and analyzes the psychological activity process of the translators, verifies the expression of translation psychology in the translation of ICH terms, and identifies the characteristics of students' translations. The study's subjects were non-English major sophomores with diverse academic backgrounds, randomly sampled from a representative polytechnic university in Guangdong. The findings of this study are significant for improving the quality of translations and promoting cross-cultural understanding of ICH terms. By understanding the psychological factors involved in the translation of ICH terms, we can overcome language and cultural barriers and accurately convey the rich cultural heritage embodied in these terms.


Flea-borne rickettsioses: ecologic considerations.
Ecologic and economic factors, as well as changes in human behavior, have resulted in the emergence of new and the reemergence of existing but forgotten infectious diseases during the past 20 years. Flea-borne disease organisms (e.g., Yersinia pestis, Rickettsia typhi, R. felis, and Bartonella henselae) are widely distributed throughout the world in endemic-disease foci, where components of the enzootic cycle are present. However, flea-borne diseases could reemerge in epidemic form because of changes in vector-host ecology due to environmental and human behavior modification. The changing ecology of murine typhus in southern California and Texas over the past 30 years is a good example of urban and suburban expansion affecting infectious disease outbreaks. In these areas, the classic rat-flea-rat cycle of R. typhi has been replaced by a peridomestic animal cycle involving, e.g., free-ranging cats, dogs, and opossums and their fleas. In addition to the vector-host components of the murine typhus cycle, we have uncovered a second typhuslike rickettsia, R. felis. This agent was identified from the blood of a hospitalized febrile patient and from opossums and their fleas. We reviewed the ecology of R. typhi and R. felis and present recent data relevant to the vector biology, immunology, and molecular characterization and phylogeny of flea-borne rickettsioses.


Not a slippery slope or sudden subversion: German medicine and national socialism in 1933.
The history of medicine this century is darkened by the downfall of the German medical profession, exposed during the doctors' trial at Nuremberg in 1946. Relying largely on documents published during 1933 in German medical journals, this paper examines two widely accepted notions of those events, metaphorically termed "slippery slope" and "sudden subversion." The first connotes a gradual slide over infinitesimal steps until, suddenly, all footing is lost; the second conveys forced take over of the profession's leadership and values. Both concepts imply that the medical profession itself became the victim of circumstances. The slippery slope concept is a prominent figure of argument in the current debate on bioethics. The evidence presented here, however, strongly suggests that the German medical community set its own course in 1933. In some respects this course even outpaced the new government, which had to rein in the profession's eager pursuit of enforced eugenic sterilizations. In 1933 the convergence of political, scientific, and economic forces dramatically changed the relationship between the medical community and the government. That same convergence is occurring again and must be approached with great caution if medicine is to remain focused on the preservation of physical and medical integrity.


Biomarkers of cerebral injury in cardiac surgery.
The present study aims to reveal the changing patterns of cerebral biomarkers and the underlying predictive factors as a consequence of cardiac surgery. Literature retrieval of articles on S100 and S100B of recent 20 years were made in the MEDLINE database, Highwire Press and Google search. Quantitative data of S100 and S100B along with neuron-specific enolase were carefully screened, collected and statistically analyzed. The biomarkers appeared earlier and lasted longer in the cerebrospinal fluid than in the serum. All three biomarkers exhibited a similar kinetic trend in the bloodstream, reaching a peak value at the end of cardiopulmonary bypass (CPB). In adults, serum biomarkers may recover to normal earlier than in pediatrics undergoing a cardiac surgery. The patients undergoing off-pump surgery had the minimal elevations of cerebral biomarkers comparing to all other cardiac surgeries under CPB, low core temperature and/or hypothermic circulatory arrest. In patients with pre- or postoperative neurological disorders, the biomarkers in the serum elevated even before operation and persisted longer time than those without neurological disorders. The serum concentrations of the biomarkers showed direct correlation with CPB duration and core temperature. Cardiac operations may lead to cerebral damage and blood-brain barrier changes, as a consequence of CPB and low core temperature. The cerebral biomarkers including S100, S100B and neuron-specific enolase may precisely reflect the cerebral damages in cardiac surgery. Attention has to be paid to the attenuations of cerebral damage by modifying the surgical conditions of CPB and core temperature.


How to avoid iatrogenic injuries of the biliary tract in urgent surgery? The use of indocianine green is an alternative.
Iatrogenic bile duct injury is a rare complication, although feared due to its morbidity and mortality. In urgent surgeries, its incidence can be doubled, so in selected cases we must assess the use of resources such as indocyanine green to minimize the risk of biliary or arterial lesions by allowing the correct identification of the structures. We present the case of a 57-year-old patient with acute cholecystitis who underwent laparoscopic cholecystectomy. Given the difficulty in differentiating structures in Calot's triangle, the decision was made to use indocyanine green, which identifies a very short cystic duct, thus avoiding iatrogenic bile duct injury.


Noninvasive diagnosis of deep vein thrombosis in postoperative patients.
The accuracy of noninvasive testing for the diagnosis of deep vein thrombosis (DVT) generally is less in asymptomatic patients than it is in those with symptoms suggestive of thrombosis. This is because asymptomatic DVT often is confined to the distal veins and, when it involves the proximal veins, the thrombi usually are smaller than in symptomatic patients with proximal thrombosis. Because the positive predictive value of noninvasive tests for asymptomatic DVT generally is 80% or less, abnormal results should be confirmed by venography. There are two main reasons why asymptomatic DVT is sought in the postoperative period: (1) to identify the need for full-dose anticoagulant therapy to prevent symptomatic episodes of venous thromboembolism (VTE), including fatal pulmonary embolism (this represents a form of secondary prophylaxis), and (2) to use this outcome as a surrogate for episodes of clinically important VTE in studies that are designed to evaluate methods of venous thrombosis prophylaxis. In relation to the first of these indications, evidence suggests that routine surveillance of high-risk patients to detect asymptomatic postoperative DVT does not result in improved clinical outcomes in patients who received appropriate VTE prophylaxis. In relation to the second indication, there is concern that asymptomatic VTE may not be a reliable surrogate for clinically important VTE, particularly if the effectiveness of different antithrombotic agents is being compared. Coupled with the comparatively low accuracy of noninvasive testing for asymptomatic DVT, this suggests that the results of such testing are unsuitable for the evaluation of new methods of prophylaxis in clinical trials.


Relationship between Glasgow coma scale and functional outcome.
The Glasgow Coma Scale (GCS) is routinely used in the acute care setting after traumatic brain injury (TBI) to guide decisions in triage, based on its ability to predict morbidity and mortality. Although the GCS has been previously demonstrated to predict mortality, efficacy in prediction of functional outcome has not been established. The purpose of this study was to assess the value of the acute GCS in predicting functional outcome in survivors of TBI. This study used the Multicenter National Institute on Disability and Rehabilitation Research TBI Model Systems database of 501 patients who had received acute medical care and inpatient rehabilitation within a coordinated neurotrauma program for treatment of TBI. Initial and lowest 24 hr GCS scores were correlated with the following outcome measures: the Disability Rating Scale (DRS), Rancho Los Amigos Levels of Cognitive Functioning Scale (LCFS), and cognitive and motor components of the Functional Independence Measure (FIM(SM)-COG and FIM(SM)-M). Outcome data were collected at admission to and discharge from the inpatient TBI rehabilitation unit. Correlation analysis revealed only modest, but statistically significant, relationships between initial and lowest GCS scores and outcome variables. Initial and lowest GCS score comparison with outcome demonstrated the following correlation coefficients: admission DRS, -0.25 and -0.28; discharge DRS, -0.24 and -0.24; admission LCFS, 0.31 and 0.33; discharge LCFS, 0.27 and 0.25; admission FIM-COG, 0.36 and 0.37; discharge FIM-COG, 0.23 and 0.23; admission FIM-M, 0.31 and 0.31; discharge FIM-M, 0.25 and 0.21. The GCS as a single variable may have limited value as a predictor of functional outcome.


Human embryonic stem cell registries: value, challenges and opportunities.
The accelerating pace of human embryonic stem cell (hESC) research has created an urgent need for the development of hESC registries, information repositories intended to gather, organize and disseminate hESC information. Although of enormous value to this evolving field, registries face significant challenges to their development. These challenges include addressing the legal and ethical issues surrounding hESC derivation as well as complex intellectual property concerns. In addition to these issues, registries must develop tools to efficiently gather, validate and present many different types of hESC information from a variety of sources. Given the pace and regulatory complexities of this field, it is important that registries develop cooperative mechanisms to avoid duplication and more efficiently support hESC research.


Transfer RNA labeling of Escherichia coli methionyl-tRNA transformylase.
Homogeneous methionyl-tRNA transformylase from Escherichia coli can react with periodate-treated tRNAMetf to form a Schiff's base through a free amino group (probably the epsilon-amino group of a lysine) and the 2',3'-aldehyde groups created at the 3'-terminal ribose of tRNA. This reaction is reflected by the loss of activity of the enzyme at saturating tRNA dialdehyde and upon reduction of the Schiff's base by NaBH4. the kinetics of inactivation (37 degrees C, pH 8.5) level off at 50% of the initial enzymic activity. In the presence of 2 mM cyanohydridoborate, a mild reducing agent which leaves intact the reacting aldehyde groups of oxidized tRNA but continuously reduces the Schiff's base in equilibrium, the activity of the enzyme can be destroyed by 100%, at a rate of 0.044 min-1, with the parallel covalent incorporation of close to one tRNA molecule per enzyme molecule. Selectivity of the labeling is also supported by the demonstration that, prior to Schiff's base formation, modified tRNA binds the transformylase with equilibrium constant and stoichiometry in good agreement with those for the active binding of unmodified tRNa. Moreover intact tRNA competes for the inactivation by the dialdehyde derivative with an affinity constant identical to that for its active binding to the enzyme


Covariance pattern mixture models: Eliminating random effects to improve convergence and performance.
Growth mixture models (GMMs) are prevalent for modeling unknown population heterogeneity via distinct latent classes. However, GMMs are riddled with convergence issues, often requiring researchers to atheoretically alter the model with cross-class constraints simply to obtain convergence. We discuss how within-class random effects in GMMs exacerbate convergence issues, even though these random effects rarely help answer typical research questions. That is, latent classes provide a discretization of continuous random effects, so including additional random effects within latent classes can unnecessarily complicate the model. These random effects are commonly included in order to properly specify the marginal covariance; however, random effects are inefficient for patterning a covariance matrix, resulting in estimation issues. Such a goal can be achieved more simply through covariance pattern models, which we extend to the mixture model context in this article (covariance pattern mixture models, or CPMMs). We provide evidence from theory, simulation, and an empirical example showing that employing CPMMs (even if they are misspecified) instead of GMMs can circumvent the computational difficulties that can plague GMMs, without sacrificing the ability to answer the types of questions commonly asked in empirical studies. Our results show the advantages of CPMMs with respect to improved class enumeration and less biased class-specific growth trajectories, in addition to their vastly improved convergence rates. The results also show that constraining the covariance parameters across classes in order to bypass convergence issues with GMMs leads to poor results. An extensive software appendix is included to assist researchers in running CPMMs in Mplus.


Evaluation of the "learning curve" for left and right radial approach during percutaneous coronary procedures.
The transradial approach for percutaneous coronary procedures may be effectively performed through the right radial approach (RRA) or left radial approach (LRA) after an appropriate "learning curve." However, studies evaluating the "learning curve" for RRA and LRA are lacking. In the Transradial Approach (Left vs Right) and Procedural Times During Percutaneous Coronary Procedures (TALENT) study, which randomized 1,540 patients to the RRA or LRA, transradial procedures were performed by either seniors or fellows. Diagnostic procedures performed by fellows were divided into 3 stages: 0 to 100 procedures (stage 1), 101 to 200 procedures (stage 2), and >200 procedures (sage 3). The primary end point of the study was fluoroscopy time during the 3 stages. Six fellows performed 532 procedures, 260 through the RRA and 272 through the LRA. During the training period, fellows showed a progressive significant reduction in fluoroscopy time for the LRA over the 3 stages (stage 1: 258 seconds, interquartile range [IQR] 138 to 377; stage 2: 198 seconds, IQR 126.5 to 375; stage 3: 142 seconds, IQR 95 to 325; p = 0.003), whereas for the RRA, only a slight and nonsignificant reduction in fluoroscopy time was observed (stage 1: 271 seconds, IQR 186 to 364; stage 2: 240 seconds, IQR 156 to 395; stage 3: 218.5 seconds, IQR 145.5 to 300.5; p = 0.20). Cannulation time was progressively reduced over the time for the 2 radial approaches: during stage 1, <40% of procedures required ≤3 minutes for radial cannulation, whereas at stage 3, radial cannulation time was ≤3 minutes in >60% of procedures (p <0.0001). In conclusion, the LRA is associated with a shorter learning curve compared to the RRA.


The flare up of catastrophic antiphospholipid syndrome: a report of an immunosuppressive withdrawal-induced case.
Antiphospholipid syndrome (APS) is a systemic disease that causes venous and arterial thrombosis in virtually any organ. Sometimes it is complicated into pulmonary infarction and cavitation, pulmonary hypertension, and catastrophic course with high morbidity and mortality. The present case is a 35-year-old woman with one episode of postpartum deep veins thrombosis (DVT) 12 years earlier and the second one after the second labor two years later. In spite of usual therapy for each episode of DVT, the condition had progressed into severe pulmonary hypertension. The diagnosis of primary APL syndrome was confirmed four years ago. She had been on warfarin, low dose of steroid, and azathioprine since the diagnosis of APL syndrome. After one year treatment with steroid and azathiprine the patient showed progressive well being; however, because of hyperglycemia the steroid tapered and discontinued. She had several attacks of paroxismal atrial tachycardia in the last year. On the last time, she presented with severe dyspnea, hemoptesis, and lower limbs edema. Chest radiography and Lung CT scan demonsterated the presence of lung cavitations. Because of high suspicious for fungal pulmonary infection, azathioprine was also discontinued. However, constellation renal failure, hemodynamic instability, and confusion caused the patient to succumb to death. The definite diagnosis of lung cavitations was not obtained.


Synthesis of isomaltooligosaccharides using 4-O-α-d-isomaltooligosaccharylmaltooligosaccharide 1,4-α-isomaltooligosaccharohydrolase.
Isomaltooligosaccharides (IMOs), including isomaltose, are valuable oligosaccharides, and the development of methods to synthesize high-purity IMOs has long been underway. We recently discovered a novel enzyme, 4-O-α-d-isomaltooligosaccharylmaltooligosaccharide 1,4-α-isomaltooligosaccharohydrolase (IMM-4IH), that showed promise for improving the synthesis process. In this study, we establish methods for synthesizing isomaltose and IMOs consisting of a variety of degrees of polymerization from starch using IMM-4IH. With 5% substrate, by combining IMM-4IH with 1,4-α-glucan 6-α-glucosyltransferase from Bacillus globisporus N75, the yield of isomaltose was 63.0%; incorporating isoamylase and cyclomaltodextrin glucanotransferase increased the yield to 75.3%. On the other hand, by combining IMM-4IH with 1,4-α-glucan 6-α-glucosyltransferase from Paenibacillus sp. PP710, IMOs were synthesized. The inclusion of isoamylase and α-amylase led to the 136 mM IMOs, consisting of oligosaccharides from isomaltose to isomaltodecaose, from 10% starch. The development of these efficient methods will be an important contribution to the industrial production of IMOs.


Why Men Fare Worse: A Mixed-Methods Study Examining Gender Differences in Nursing Home Resident Quality of Life.
Despite research documenting gender differences in numerous outcomes in later life, we know little about gender differences in quality of life (QoL) for nursing home (NH) residents. This study examines the relationship between gender and residents' QoL, including possible reasons for differences observed. We used a mixed-methods design including surveys with a random sample of Minnesota NH residents using a multidimensional measure of QoL (n = 9,852), resident clinical data, facility-level characteristics (n = 364), interviews with residents (n = 64), and participant observations. We used linear mixed models and thematic analysis of resident interviews and observations to examine possible gender-related differences in residents' QoL. After controlling for individual and facility characteristics, men reported lower overall QoL than women, including significantly lower ratings in several QoL domains. In interviews, men noted being less satisfied with activities than women, having fewer friends, and being less able to rely on family for support. Some women described the NH as a place of respite, but men more often described being dissatisfied with life in the NH and undesirable for long-term living. Our observations were consistent with interview findings but provide additional nuances, such as that some men organized their own social groups. Our findings suggest that men and women experience QoL differently in NHs, with men reporting lower QoL in several domains. Tailoring more activities for men and finding ways to strengthen relationships for men in NHs could help reduce the gender differences in QoL we observed.


Effects of dietary fatty acid content on humeral cartilage and bone structure in a mouse model of diet-induced obesity.
Obesity is a primary risk factor for osteoarthritis (OA), and previous studies have shown that dietary content may play an important role in the pathogenesis of cartilage and bone in knee OA. Several previous studies have shown that the ratio of ω-3 polyunsaturated fatty acids (PUFAs), ω-6 PUFAs, and saturated fatty acids can significantly influence bone structure and OA progression. However, the influence of obesity or dietary fatty acid content on shoulder OA is not well understood. The goal of this study was to investigate the role of dietary fatty acid content on bone and cartilage structure in the mouse shoulder in a model of diet-induced obesity. For 24 weeks, mice were fed control or high-fat diets supplemented with ω-3 PUFAs, ω-6 PUFAs, or saturated fatty acids. The humeral heads were analyzed for bone morphometry and mineral density by microCT. Cartilage structure and joint synovitis were determined by histological grading, and microscale mechanical properties of the cartilage extracellular and pericellular matrices were quantified using atomic force microscopy. Diet-induced obesity significantly altered bone morphology and mineral density in a manner that was dependent on dietary free fatty acid content. In general, high-fat diet groups showed decreased bone quality, with the ω-3 diet being partially protective. Cartilage mechanical properties and OA scores showed no changes with obesity or diet. These findings are consistent with clinical literature showing little if any relationship between obesity and shoulder OA (unlike knee OA), but suggest that diet-induced obesity may influence other joint tissues. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res.


Basement Membrane Matrix Promotes the Efficiency of Hair Follicle Reconstruction In Vivo and Is a Cell Delivery Vehicle for Follicle Regeneration.
The efficiency of hair follicle (HF) reconstruction is decreased by extensive apoptotic remodeling that occurs soon after grafting. To evaluate a basement membrane matrix (matrix) to improve the efficiency of HF reconstruction and serve as a cell delivery vehicle. Newborn mouse skin cells were suspended in a matrix and transplanted in a chamber assay. The viability and proliferation of mouse dermal papilla cells seeded in the matrix were tested. Dermal papilla cells and epidermal cells seeded in matrix sheets were grafted into nude mice to observe hair formation. The matrix significantly shortened the time to hair formation. The first hair shafts appeared within the matrix at 17.67 ± 1.21 days versus 23.00 ± 1.41 days for Dulbecco's modified Eagle medium controls. There was a significant difference (p < .05) in the number of newly formed hairs in areas of reconstructed skin with the matrix (100 μL) grafts (323 ± 12) versus controls (276 ± 11). Dermal papilla cells were successfully cultured in the matrix, and hair formation was dense when the matrix was used as a cell delivery vehicle for follicle reconstruction. The matrix improved the efficiency of HF reconstruction and was a suitable delivery vehicle of cells for HF engineering.


Plasma rich in growth factors had limited effect on early bone formation in extraction sockets: Response to "Anitua, E., Alkhraisat, M.H. & Orive, G. (2013) Letter to the Editor: Rigorous methodology is the school of coherent conclusions in science. European Journal of Oral Implantology 6: 9-11.".
To address the criticisms raised by Anitua et al. (European Journal of Oral Implantology, 6, 2013, 9-11) to the article "Plasma Rich in Growth Factors (PRGF) in Human Post-Extraction Sockets: an Histological and Histomorphometric Study.", recently published by Farina and colleagues (Clinical Oral Implants Research 2012; doi: 10.1111/clr.12033). All the methodological aspects criticized in the letter by Anitua et al. were thoroughly reconsidered and discussed in a structured short communication. When indicated, pertinent, additional material was included to reinforce our considerations. As most clinical studies in implant dentistry, including previous studies evaluating the efficacy/effectiveness of PRGF, the study by Farina et al. has some limitations. However, it is currently the only published controlled trial using quantitative parameters related to PRGF-induced early bone formation. Despite all limitations, the results of the study by Farina et al., which were based on different quantitative parameters (micro-CT scan, immunohistochemical markers of wound healing and bone deposition), indicated a limited effect of PRGF on early bone formation in extraction sockets.


Applanation tonometry in the conscious cynomolgus monkey (Macaca fascicularis).
Four monkeys (Macaca fascicularis) were trained to tolerate applanation tonometry while fully conscious. The tonometer used was the Draeger tonometer with an applanation area of 3.06 diameter, but with a reduced application surface (4 mm instead of 6.8 mm diameter). The reduction did not change the calibration and is applicable in this monkey species without any correction factor. The mean intraocular pressure determined was 19.12 mmHg.


Rationale for treatment of cancer with calcium antagonists. A review.
Calcium antagonists are broad spectrum antineoplastic agents capable of influencing favorably the course of cancer by inhibiting growth factor-initiated cell activation, blocking metastasis, or reversing inherent or acquired resistance to chemotherapeutic drugs. Calcium antagonists constitute an attractive category of agents for clinical trials in humans because their effects on malignancy in vivo in experimental animals are dramatic, and because there are several such agents on the market that can be adapted readily for such studies. However, an appreciation of the mechanisms of action of these drugs on cancer is essential for selection of an experimental design for therapeutic trials that will test these mechanisms adequately. The purpose of this review is to summarize available information on effects of calcium antagonists on cancer to facilitate incorporation of such drugs into therapeutic trials in human malignancy.


[The field of Collective Health in the courses perspective].
This article discusses the field of Collective Health from the disciplines that structure the curriculum of the Collective Health area postgraduate courses in 2006. From the database of programs, including master's, doctoral and professional masters were classified 1,171 subjects, in what we call disciplinary fields, linking this to the P. Bourdieu's notion of champ (field). Twenty-eight disciplinary fields were described, in three large groups. It was concluded that the field of collective health has a wide range of disciplinary fields, especially the frequency: Epidemiology (16%), Humanities and Social Sciences in Health (7.9%); Curriculum (7.9%); Systems and Health Services (7.4%), Environment and Health (7.2%) Health Policies (6.3%).


Aloe vera toxic effects: expression of inducible nitric oxide synthase (iNOS) in testis of Wistar rat.
Nitric oxide (NO), a product of inducible nitric oxide synthase (iNOS), contributes in germ cell apoptosis. This study was aimed to evaluate the effects of Aloe vera gel (AVG) on male Wistar rat reproductive organ, serum NO level, and expression of iNOS gene in leydig cells. Adult male Wistar rats (n=36) were used for experiments in three groups. The experimental groups were orally administered with the AVG extract solution once-daily as follow: 150 mg.kg(-1); group A, 300 mg.kg(-1); group B, and only normal saline; group C (control group). They were mated with untreated females and the reproductive and chemical parameters were assessed for each group, including semen quality, serum testosterone, sperm fertility, gonad and body weight, serum NO concentration (by the Griess method), and iNOS gene expression (using RT-PCR). The testes weight, serum testosterone, as well as sperm count and fertility of the AVG treated groups were significantly reduced when compared to the control (P<0.001). Concentration of serum NO was significantly increased (37.1±4.63 µM) in the administrated group with higher AVG concentration, compared to the control group (P<0.001; 10.19±0.87 µM); however, iNOS mRNA expression was increased in the treated animals (P<0.001). iNOS may play a functional role in spermatogenesis via apoptosis, reducing sperm count, but further studies are needed to illustrate the mechanisms by which AVG exerts its negative effects on spermatogenesis and sperm quality.


The PCSK9 revolution: Current status, controversies, and future directions.
Proprotein Convertase Subtilisin/Kexin Type 9 (PCSK9) has revolutionized our understanding of cholesterol homeostasis and added to our arsenal against atherosclerotic cardiovascular disease (ASCVD). In a span of approximately 15 years, PCSK9 has morphed from an esoteric and rare cause of familial hypercholesterolemia (FH) into the most efficient cholesterol-lowering target ever known, with the completion of two large scale cardiovascular outcome trials showing positive results. Current Food and Drug Administration (FDA) approved modalities to inhibit PCSK9 are in the form of monoclonal antibodies which display an unparalleled degree of low-density lipoprotein cholesterol (LDL-C) lowering and expand upon the notion that lower LDL-C is better for ASCVD risk reduction. However, the accelerated pace of discovery and therapeutic development has left large gaps in our knowledge regarding the physiology and function of PCSK9. The aim of this review is to provide context to the discovery, history, treatment and current status of PCSK9 and its therapeutic inhibitors and highlight areas of controversy and future directions.


Molecular and functional characterization of a cDNA encoding fructan:fructan 6G-fructosyltransferase (6G-FFT)/fructan:fructan 1-fructosyltransferase (1-FFT) from perennial ryegrass (Lolium perenne L.).
Fructans are the main storage compound in Lolium perenne. To account for the prevailing neokestose-based fructan synthesis in this species, a cDNA library of L. perenne was screened by using the onion (Allium cepa) fructan:fructan 6G-fructosyltransferase (6G-FFT) as a probe. A full length Lp6G-FFT clone was isolated with significant homologies to vacuolar type fructosyltransferases and invertases. The functionality of the cDNA was tested by heterologous expression in Pichia pastoris. The recombinant protein demonstrated both 6G-FFT and fructan:fructan 1-fructosyltransferase activities (1-FFT) with a maximum 6G-FFT/1-FFT ratio of two. The activity of 6G-FFT was investigated with respect to developmental stage, tissue distribution, and alterations in carbohydrate status expression and compared to sucrose:sucrose 1-fructosyltransferase (1-SST). Lp6G-FFT and Lp1-SST were predominantly expressed in the basal part of elongating leaves and leaf sheaths. Expression of both genes declined along the leaf axis, in parallel with the spatial occurrence of fructan and fructosyltransferase activities. Surprisingly, Lp6G-FFT was highly expressed in photosynthetically active tissues where very low extractable fructosyltransferase activity and fructan amounts were detected, suggesting a post-transcriptional regulation of expression. Lp6G-FFT gene expression increased only in elongating leaves following similar increases of sucrose content in blades, sheaths, and elongating leaf bases. Regulation of Lp6G-FFT gene expression depends on the tissue according to its sink-source status.


Learning With Noisy Labels via Self-Reweighting From Class Centroids.
Although deep neural networks have been proved effective in many applications, they are data hungry, and training deep models often requires laboriously labeled data. However, when labeled data contain erroneous labels, they often lead to model performance degradation. A common solution is to assign each sample with a dynamic weight during optimization, and the weight is adjusted in accordance with the loss. However, those weights are usually unreliable since they are measured by the losses of corrupted labels. Thus, this scheme might impede the discriminative ability of neural networks trained on noisy data. To address this issue, we propose a novel reweighting method, dubbed self-reweighting from class centroids (SRCC), by assigning sample weights based on the similarities between the samples and our online learned class centroids. Since we exploit statistical class centers in the image feature space to reweight data samples in learning, our method is robust to noise caused by corrupted labels. In addition, even after reweighting the noisy data, the decision boundaries might still suffer distortions. Thus, we leverage mixed inputs that are generated by linearly interpolating two random images and their labels to further regularize the boundaries. We employ the learned class centroids to evaluate the confidence of our generated mixed data via measuring feature similarities. During the network optimization, the class centroids are updated as more discriminative feature representations of original images are learned. In doing so, SRCC will generate more robust weighting coefficients for noisy and mixed data and facilitates our feature representation learning in return. Extensive experiments on both the synthetic and real image recognition tasks demonstrate that our method SRCC outperforms the state of the art on learning with noisy data.


Synthesis and in vitro and in vivo evaluation of an (18)F-labeled neuropeptide Y analogue for imaging of breast cancer by PET.
Imaging of Y1R expression in breast cancer is still a challenging task. Herein, we report a suitable (18)F-labeled high-molecular-weight glycopeptide for imaging of peripheral neuropeptide Y (NPY) Y1 receptor (Y1R)-positive tumors by preclinical small-animal positron emission tomography (PET). The Y1R-preferring NPY [F(7),P(34)]NPY analogue was functionalized with an alkyne-bearing propargylglycine (Pra) in position 4. The corresponding fluoroglycosylated (FGlc) peptide analogue [Pra(4)(FGlc),F(7),P(34)]NPY and its (18)F-labeled analogue were synthesized by click chemistry-based fluoroglycosylation. The radiosynthesis was performed by (18)F-fluoroglycosylation starting from the 2-triflate of the β-mannosylazide and the alkyne peptide [Pra(4),F(7),P(34)]NPY. The radiosynthesis of the(18)F-labeled analogue was optimized using a minimum amount of peptide precursor (40 nmol), proceeding with an overall radiochemical yield of 20-25% (nondecay corrected) in a total synthesis time of 75 min with specific activities of 40-70 GBq/μmol. In comparison to NPY and [F(7),P(34)]NPY, in vitro Y1R and Y2R activation studies with the cold [Pra(4)(FGlc),F(7),P(34)]NPY on stably transfected COS-7 cells displayed a high potency for the induction of Y1R-specific inositol accumulation (pEC50 = 8.5 ± 0.1), whereas the potency at Y2R was significantly decreased. Internalization studies on stably transfected HEK293 cells confirmed a strong glycopeptide-mediated Y1R internalization and a substantial Y1R subtype selectivity over Y2R. In vitro autoradiography with Y1R-positive MCF-7 tumor tissue slices indicated high specific binding of the (18)F-labeled glycopeptide, when binding was reduced by 95% ([Pra(4),F(7),P(34)]NPY) and by 86% (BIBP3226 Y1R antagonist) in competition studies. Biodistribution and small-animal PET studies on MCF-7 breast tumor-bearing nude mice revealed radiotracer uptake in the MCF-7 tumor of 1.8%ID/g at 20 min p.i. and 0.7%ID/g at 120 min p.i. (n = 3-4), increasing tumor-to-blood ratios from 1.2 to 2.4, and a tumor retention of 76 ± 4% (n = 4; 45-90 min p.i.). PET imaging studies with MCF-7 tumor-bearing nude mice demonstrated uptake of the (18)F-labeled glycopeptide in the tumor region at 60 min p.i., whereas only negligible tumor uptake was observed in animals injected with a nonbinding (18)F-labeled glycopeptide pendant as a measure of nonspecific binding. In conclusion, PET imaging experiments with the (18)F-labeled NPY glycopeptide revealed Y1R-specific binding uptake in MCF-7 tumors in vivo together with decreased kidney uptake compared to DOTA-derivatives of this peptide. We consider this glycopeptide to be a potent lead peptide for the design of improved (18)F-glycopeptides with shorter amino acid sequences that would further facilitate PET imaging studies of Y1R-positive breast tumors.


Diffuse intestinal T-cell lymphosarcoma in a yellow-naped Amazon parrot (Amazona ochrocephala auropalliata).
A 10-year-old, intact, female yellow-naped Amazon parrot was examined because of anemia, lymphocytic leukocytosis, regurgitation, and weight loss. A positive fecal occult blood and monoclonal globulinopathy were present. A distended proventriculus and diffusely thickened loops of small intestine with irregular luminal surfaces were identified with contrast radiography and contrast computed tomography. A micro positron emission tomography scan was performed with (18)F-fluorodeoxyglucose. Diffuse intestinal T-cell lymphosarcoma was diagnosed based on histopathology and immunohistochemistry of full thickness small intestinal biopsies. The patient was treated with a multidrug chemotherapy protocol with little to no effect. Euthanasia was elected, and intestinal lymphosarcoma was confirmed on histopathology of necropsy intestinal samples; no other organs demonstrated neoplastic infiltration. To the authors' knowledge, no reports are currently available detailing the clinical presentation or diagnosis of diffuse intestinal T-cell lymphosarcoma in any avian species.


Late open conversion after TEVAR.
The aim of this paper was to characterize indications, describe repair methods and report outcomes of patients undergoing late open conversion (LOC) after thoracic endovascular aortic repair (TEVAR). From to 1999 to 2015, 65 patients underwent redo procedures after TEVAR at our Institution. Thirty patients underwent LOC. The average time from the index TEVAR to LOC was 57 months (range 2-98 months). Indication to LOC was progression of distal aortic disease in nine patients (30%), stent-graft infection/fistulization in 10 (33.3%), retrograde dissection in 6 (20%), endoleak in 3 (10%), device migration/failure in 2 (6.7%). Left heart bypass and cerebrospinal fluid drainage were used in 22 (73.3%) and 16 (53.3%) cases, respectively. In 17 patients (56.6%) thoracic stent-graft was only partially removed and the proximal anastomosis was performed including the native aorta, the stent-graft and the surgical graft. Biological flap coverage with intercostal muscle flap was used 6 six of infected cases. Overall perioperative mortality rate was 16.6% (5 patients) - 20% in the subgroup of patients with stent-graft infection/fistulization (2 patients) and 33% in the subgroup with retrograde dissection (2 patients). Respiratory failure was the most common complication with seven cases (23.3%). Paraplegia was observed in 1 patient (3.3%). At a mean follow-up of 19 months (range, 5-63) there was 1 aortic-related death for aorto-esophageal fistula. Open conversion after TEVAR was feasible leaving in site the proximal segment of the thoracic stent-graft in most of the cases. The risk of death and morbidity was higher in case of infection, fistula and retrograde dissection.


Enhanced Deposition by Electrostatic Field-Assistance Aggravating Diesel Exhaust Aerosol Toxicity for Human Lung Cells.
Air pollution is associated with increased risk of cardiovascular and pulmonary diseases, but conventional air quality monitoring gives no information about biological consequences. Exposing human lung cells at the air-liquid interface (ALI) to ambient aerosol could help identify acute biological responses. This study investigated electrode-assisted deposition of diesel exhaust aerosol (DEA) on human lung epithelial cells (A549) in a prototype exposure chamber. A549 cells were exposed to DEA at the ALI and under submerged conditions in different electrostatic fields (EFs) and were assessed for cell viability, membrane integrity, and IL-8 secretion. Qualitative differences of the DEA and its deposition under different EFs were characterized using scanning mobility particle sizer (SMPS) measurements, transmission electron microscopy (TEM), and electron energy loss spectroscopy (EELS). Upon exposure to DEA only, cell viability decreased and membrane impairment increased for cells at the ALI; submerged cells were unaffected. These responses were enhanced upon application of an EF, as was DEA deposition. No adverse effects were observed for filtered DEA or air only, confirming particle-induced responses. The prototype exposure chamber proved suitable for testing DEA-induced biological responses of cells at the ALI using electrode-assisted deposition and may be useful for analysis of other air pollutants.


Intramuscular metabolism during low-intensity resistance exercise with blood flow restriction.
Although recent studies have reported that low-intensity resistance training with blood flow restriction could stress the muscle effectively and provide rapid muscle hypertrophy and strength gain equivalent to those of high-intensity resistance training, the exact mechanism and its generality have not yet been clarified. We investigated the intramuscular metabolism during low-intensity resistance exercise with blood flow restriction and compared it with that of high-intensity and low-intensity resistance exercises without blood flow restriction using (31)P-magnetic resonance spectroscopy. Twenty-six healthy subjects (22 +/- 4 yr) participated and performed unilateral plantar flexion (30 repetitions/min) for 2 min. Protocols were as follows: low-intensity exercise (L) using a load of 20% of one-repetition maximum (1 RM), L with blood flow restriction (LR), and high-intensity exercise using 65% 1 RM (H). Intramuscular phosphocreatine (PCr) and diprotonated phosphate (H(2)PO(4)(-)) levels and intramuscular pH at rest and during exercise were obtained. We found that the PCr depletion, the H(2)PO(4)(-) increase, and the intramuscular pH decrease during LR were significantly greater than those in L (P < 0.001); however, those in LR were significantly lower than those in H (P < 0.001). The recruitment of fast-twitch fiber evaluated by inorganic phosphate splitting occurred in only 31% of the subjects in LR, compared with 70% in H. In conclusion, the metabolic stress in skeletal muscle during low-intensity resistance exercise was significantly increased by applying blood flow restriction, but did not generally reach that during high-intensity resistance exercise. This new method of resistance training needs to be examined for optimization of the protocol to reach equivalence with high-intensity resistance training.


Structural basis for activation of class Ib ribonucleotide reductase.
The class Ib ribonucleotide reductase of Escherichia coli can initiate reduction of nucleotides to deoxynucleotides with either a Mn(III)2-tyrosyl radical (Y•) or a Fe(III)2-Y• cofactor in the NrdF subunit. Whereas Fe(III)2-Y• can self-assemble from Fe(II)2-NrdF and O2, activation of Mn(II)2-NrdF requires a reduced flavoprotein, NrdI, proposed to form the oxidant for cofactor assembly by reduction of O2. The crystal structures reported here of E. coli Mn(II)2-NrdF and Fe(II)2-NrdF reveal different coordination environments, suggesting distinct initial binding sites for the oxidants during cofactor activation. In the structures of Mn(II)2-NrdF in complex with reduced and oxidized NrdI, a continuous channel connects the NrdI flavin cofactor to the NrdF Mn(II)2 active site. Crystallographic detection of a putative peroxide in this channel supports the proposed mechanism of Mn(III)2-Y• cofactor assembly.


[Effect of cadmium on electromechanical coupling in the frog myocardium].
CdCl2 (0.2-0.5 mM) shortened the duration of action potential and reduced the tension of frog ventricular strips. CdCl2 completely blocked slow AP in partially depolarized strips and abolished both the tonic and phasic components of KCl-induced contractures. CdCl2 prevents adrenaline, NiCl2 and low-Na Ringer solution to produce tension potentiation. Cd seems to block the slow Na-Ca channels of frog myocardium and disturb intracellular Ca distribution by its ability to block protein SH-groups.


Role of bone scintigraphy in the early diagnosis of discitis.
Nine cases of recorded discitis are presented, in which scintigraphy played an important part in the diagnosis. The patients (five men (aged 21-75 years) and four women (aged 40-73 years)) had a history of back pain varying in duration from two days to three months. Final diagnosis was confirmed microbiologically (seven patients) or radiographically (two patients). Bone scintigraphy was a valuable diagnostic procedure for discitis with earlier detection than plain radiography in three patients and similar initial detection to that of third generation computed tomography. Single photon emission computed tomographic imaging increased diagnostic confidence by indicating the involvement of the adjacent vertebral bodies rather than of the pedicles or spinous processes.


Phase I, open-label, single-dose study to evaluate the pharmacokinetics and safety of telbivudine in children and adolescents with chronic hepatitis B.
Telbivudine is a nucleoside analogue that has been approved for the treatment of chronic hepatitis B virus (HBV) infection in adults at 600 mg/day. We conducted a phase I, open-label, first-in-pediatrics study to investigate the safety and pharmacokinetics of a single dose of telbivudine in HBV-infected children and adolescents. Eligible patients were enrolled sequentially from older to younger groups, with evaluation of safety and available pharmacokinetic data after each stratum. Adolescent patients (>12 to 18 years) received a single dose of 600 mg telbivudine as an oral solution, while children aged 2 to 12 years received a single dose of 15 or 25 mg/kg of body weight up to a maximum of 600 mg. Telbivudine was well tolerated; all adverse events were mild, and none occurred in more than one patient. The plasma telbivudine concentration-versus-time profiles in adolescents given 600 mg were similar to the mean profile of healthy adults receiving the same oral dose. Children aged 2 to <6 and 6 to 12 years receiving a single 15-mg/kg dose showed similar plasma exposures. To predict the steady-state exposure, plasma concentration-versus-time profiles for patients aged 2 to 12 years (15 mg/kg) and >12 to 18 years (600 mg) were fitted to a two-compartment 1st-order, microconstant, lag time, 1st-order elimination pharmacokinetic (PK) model. This analysis predicted the following dosages to mimic exposures in healthy adults receiving 600 mg/day: 20 mg/kg/day for children 2 to 12 years and 600 mg/day for adolescents. Studies are ongoing to evaluate the efficacy of the recommended dose in pediatric patients. (This study has been registered at ClinicalTrials.gov under registration no. NCT00907894.).


Rotational coherence imaging and control for CN molecules through time-frequency resolved coherent anti-Stokes Raman scattering.
Numerical wave packet simulations are performed for studying coherent anti-Stokes Raman scattering (CARS) for CN radicals. Electronic coherence is created by femtosecond laser pulses between the X(2)Σ and B(2)Σ states. Due to the large energy separation of vibrational states, the wave packets are superpositions of rotational states only. This allows for a specially detailed inspection of the second- and third-order coherences by a two-dimensional imaging approach. We present the time-frequency domain images to illustrate the intra- and intermolecular interferences, and discuss the procedure to rationally control and experimentally detect the interferograms in solid Xe environment.


Hemodynamic effects of ionized calcium in horses anesthetized with halothane or isoflurane.
To evaluate the effects of halothane and isoflurane on cardiovascular function and serum total and ionized calcium concentrations in horses, and to determine whether administration of calcium gluconate would attenuate these effects. 6 clinically normal adult Thoroughbreds. Catheters were inserted for measurement of arterial blood pressures, pulmonary arterial blood pressures, right ventricular pressure (for determination of myocardial contractility), right atrial pressure, and cardiac output and for collection of arterial blood samples. Anesthesia was then induced with xylazine hydrochloride and ketamine hydrochloride and maintained with halothane or isoflurane. An i.v. infusion of calcium gluconate was begun 75 minutes after anesthetic induction; dosage of calcium gluconate was 0.1 mg/kg of body weight/min for the first 15 minutes, 0.2 mg/kg/min for the next 15 minutes, and 0.4 mg/kg/min for an additional 15 minutes. Data were collected before, during, and after administration of calcium gluconate. Halothane and isoflurane decreased myocardial contractility, cardiac index, and mean arterial pressure, but halothane caused greater depression than isoflurane. Calcium gluconate attenuated the anesthetic-induced depression in cardiac index, stroke index, and maximal rate of increase in right ventricular pressure when horses were anesthetized with isoflurane. When horses were anesthetized with halothane, a higher dosage of calcium gluconate was required to attenuate the depression in stroke index and maximal rate of increase in right ventricular pressure; cardiac index was not changed with calcium administration. I.v. administration of calcium gluconate may support myocardial function in horses anesthetized with isoflurane.


Non-Radiographic Axial Spondyloarthritis.
The term non-radiographic axial spondyloarthritis (nrAxSpA) was coined for patients who have a clinical picture of ankylosing spondylitis (AS) but do not exhibit radiographic sacroiliitis. The ASAS classification criteria for nrAxSpA, ensuring the recruitment of homogenous study cohorts, were accepted in 2009, although the respective diagnostic criteria for daily clinical practice have not yet been developed. The clinical diagnosis should be based on the composite of clinical symptoms and signs of the disease, HLA B27 status, and magnetic resonance imaging (MRI) of sacroiliac joints. Notably, a negative MRI or HLA B27 does not exclude the diagnosis in patients with a high clinical suspicion for nrAxSpA. The prevalence of nrAxSpA is similar to that of AS, but the former has a higher female preponderance. The rate of progression of nrAxSpA to the radiographic stage of disease (AS) ranges from 10% to 20% over 2 years. Current treatment strategies for nrAxSpA are the same as for AS and include non-steroidal anti-inflammatory drugs and inhibitors of tumor necrosis factor-alpha. While this review summarizes the current achievements in the field of nrAxSpA, further understanding of the epidemiology and natural history of the disease and, particularly, mechanisms of inflammation and subsequent new bone formation is essential for the development of new treatment strategies for nrAxSpA patients.


Effects of radiofrequency electromagnetic fields on the human nervous system.
The effects of exposure to radiofrequency electromagnetic fields (EMF), specifically related to the use of mobile telephones, on the nervous system in humans have been the subject of a large number of experimental studies in recent years. There is some evidence of an effect of exposure to a Global System for Mobile Telecommunication (GSM)-type signal on the spontaneous electroencephalogram (EEG). This is not corroborated, however, by the results from studies on evoked potentials. Although there is some evidence emerging that there may be an effect of exposure to a GSM-type signal on sleep EEG, results are still variable. In summary, exposure to a GSM-type signal may result in minor effects on brain activity, but such changes have never been found to relate to any adverse health effects. No consistent significant effects on cognitive performance in adults have been observed. If anything, any effect is small and exposure seems to improve performance. Effects in children did not differ from those in healthy adults. Studies on auditory and vestibular function are more unequivocal: neither hearing nor the sense of balance is influenced by short-term exposure to mobile phone signals. Subjective symptoms over a wide range, including headaches and migraine, fatigue, and skin itch, have been attributed to various radiofrequency sources both at home and at work. However, in provocation studies a causal relation between EMF exposure and symptoms has never been demonstrated. There are clear indications, however, that psychological factors such as the conscious expectation of effect may play an important role in this condition.


A multicenter analysis of safety and outcome of removal of a fully covered self-expandable metal stent during ERCP.
Fully covered self-expandable metal stents (FCSEMSs) have been used for palliation of both malignant and benign biliary strictures. Limited data are available about safety and outcome of endoscopic removal of these stents. To evaluate safety and efficacy of endoscopic removal of FCSEMSs. Retrospective review of patients who underwent endoscopic removal of Viabil FCSEMSs. Four centers with experience in using FCSEMSs. Thirty-seven patients who had stents endoscopically removed. ERCP with endoscopic removal of FCSEMSs. Feasibility, safety, and complications associated with endoscopic removal of FCSEMS. All 37 stent removal attempts were successful and were achieved without difficulty. Indwelling stent-related complications occurred in 4 of 37 patients, including secondary strictures in 3 and minor bile leak in 1. Two of 3 secondary strictures occurred at the distal stent margin of oversized intraductal stents, and another stricture occurred at a proximal stent margin of an oversized transpapillary stent. One case of minor confined intratumoral bile leak also occurred in a patient with metastatic urothelial cancer of the bile duct. All of these cases were successfully treated with repeat stenting and resolved without sequelae. No free perforations or significant bleeding occurred. Retrospective study. Endoscopic removal of Viabil FCSEMSs placed for benign or malignant conditions is feasible and easily accomplished. Secondary strictures may be found at the time of stent removal or with a delayed presentation in patients with oversized stents who may require repeat stent placement.


Intraradicular Dentinal Changes Following Fiber Postremoval: Micro-computed Tomography Analysis.
Excessive dentine wear during postremoval can occur due to the adhesion between the cement and the post. The aim is to evaluate dentine wear and microcrack occurrence following the removal of intraradicular fiberglass posts that were previously cemented with two types of resin cement. Distal roots (n = 38) of lower molars were instrumented, filled, prepared for intraradicular postinstallation, and divided into two groups according to the cement used for the posts: RelyX ARC or RelyX U200. The posts were removed through abrasion using ultrasonic diamond tips, after which the degrees of dentine wear (volume/mm3) and microcrack formation were assessed using microcomputed tomographic imaging. The Mann-Whitney U (P ≤ 0.05) and Chi-squared tests (P ≤ 0.05) were used. After the removal of the posts, no statistically significant difference in root canal volume or microcrack occurrence was observed between the groups (P > 0.05). The procedure produced significant dentine wear regardless of the type of cement used (P < 0.05). The removal of fiberglass posts caused extensive dentine wear but did not influence microcrack occurrence. The type of resin cement used did not affect the dentine loss or microcrack appearance that resulted from fiberglass postremoval.


Modulation of myelin basic protein-induced aggregation and fusion of liposomes by cholesterol, aliphatic aldehydes and alkanes.
The effect of cholesterol on myelin basic protein-induced aggregation of zwitterionic phospholipid vesicles was studied by turbidimetry, quasi-elastic light scattering and centrifugation techniques. Without cholesterol, the degree of vesicle aggregation caused by myelin basic protein is relatively low and is only slightly increased using cholesterol concentrations up to approx. 25-30 mol%. When the cholesterol content in the bilayer exceeds approx. 30 mol%, there is a dramatic increase in the susceptibility of the vesicles to aggregation in the presence of myelin basic protein. Palmitoyl aldehyde and eicosane, substances resembling products of lipid degradation, increase myelin basic protein promoted fusion of vesicles. The fusion is accompanied by increased leakage of entrapped carboxyfluorescein. In the presence of cholesterol, myelin basic protein-induced fusion of the liposomes becomes much more sensitive to the presence of aliphatic aldehydes or alkanes. The results suggest that cholesterol has an important role in promoting membrane adhesion in biological systems but these structures become unstable in the presence of small amounts of products of lipid degradation. The findings have important implications to the understanding of the stability of the myelin membrane.


Characterization of a novel, dominant negative KCNJ2 mutation associated with Andersen-Tawil syndrome.
Andersen-Tawil syndrome is characterized by periodic paralysis, ventricular ectopy, and dysmorphic features. Approximately 60% of patients exhibit loss-of-function mutations in KCNJ2, which encodes the inwardly rectifying K(+) channel pore forming subunit Kir2.1. Here, we report the identification of a novel KCNJ2 mutation (G211T), resulting in the amino acid substitution D71Y, in a patient presenting with signs and symptoms of Andersen-Tawil syndrome. The functional properties of the mutant subunit were characterized using voltage-clamp experiments on transiently transfected HEK-293 cells and neonatal mouse ventricular myocytes. Whole-cell current recordings of transfected HEK-293 cells demonstrated that the mutant protein Kir2.1-D71Y fails to form functional ion channels when expressed alone, but co-assembles with wild-type Kir2.1 subunits and suppresses wild-type subunit function. Further analysis revealed that current suppression requires at least two mutant subunits per channel. The D71Y mutation does not measurably affect the membrane trafficking of either the mutant or the wild-type subunit or alter the kinetic properties of the currents. Additional experiments revealed that expression of the mutant subunit suppresses native I(K1) in neonatal mouse ventricular myocytes. Simulations predict that the D71Y mutation in human ventricular myocytes will result in a mild prolongation of the action potential and potentially increase cell excitability. These experiments indicate that the Kir2.1-D71Y mutant protein functions as a dominant negative subunit resulting in reduced inwardly rectifying K(+) current amplitudes and altered cellular excitability in patients with Andersen-Tawil syndrome.


Angiogenesis and parasitic helminth-associated neovascularization.
Successful metazoan parasitism, among many other factors, requires a supply of nutrients and the removal of waste products. There is a prerequisite for a parasite-defined vasculature. The angiogenic mechanism(s) involved presumably depend on the characteristics of the tissue- and vascular system-dwelling, parasitic helminths. Simplistically, 2 possibilities or a combination of both have been considered in this review. The multifactorial induction of parasitic helminth-associated neovascularization could arise through, either a host-, a parasite- or a host-/parasite-dependent, angiogenic switch. Most studies appear to support the first and third hypotheses, but evidence exists for the intrahepatic cestode Echinococcus multilocularis, the free-living nematode Caenorhabditis elegans and the intravascular trematode Schistosoma mansoni for the second inference. In contrast, the nematode anti-coagulant protein NAPc2 from adult Ancylostoma caninum is also an anti-angiogenic factor.


Sensory neurons in the human jugular ganglion.
The jugular ganglion (JG) contains sensory neurons of the vagus nerve which innervate somatic and visceral structures in cranial and cervical regions. In this study, the number of sensory neurons in the human JG was investigated. And, the morphology of sensory neurons in the human JG and nodose ganglion (NG) was compared. The estimated number of JG neurons was 2721.8-9301.1 (average number of sensory neurons ± S.D. = 7975.1 ± 3312.8). There was no significant difference in sizes of the neuronal cell body and nucleus within the JG (cell body, 1128.8 ± 99.7 μ m2; nucleus, 127.7 ± 20.8 μ m2) and NG (cell body, 963.8 ± 225.7 μ m2; nucleus, 123.2 ± 32.3 μ m2). These findings indicate that most of sensory neurons show the similar morphology in the JG and NG. Our immunohistochemical method also demonstrated the distribution of ion channels, neurotransmitter agents and calcium-binding proteins in the human JG. Numerous JG neurons were immunoreactive for transient receptor potential cation channel subfamily V member 1 (TRPV1, mean ± SD = 19.9 ± 11.5 %) and calcitonin gene-related peptide (CGRP, 28.4 ± 6.7 %). A moderate number of JG neurons contained TRPV2 (12.0 ± 4.7 %), substance P (SP, 15.7 ± 6.9 %) and secreted protein, acidic and rich in cysteine-like 1 (SPARCL1, 14.6 ± 7.4 %). A few JG neurons had vesicular glutamate transporter 2 (VGLUT2, 5.6 ± 2.9 %) and parvalbumin (PV, 2.3 ± 1.4 %). SP- and TRPV2-containing JG neurons had mainly small and medium-sized cell bodies, respectively. TRPV1- and VGLUT2- containing JG neurons were small to medium-sized. CGRP- and SPARCL1-containing JG neurons were of various cell body sizes. Sensory neurons in the human JG were mostly free of vasoactive intestinal polypeptide (VIP), tyrosine hydroxylase (TH) and neuropeptide Y (NPY). In the external auditory canal skin, subepithelial nerve fibers contained TRPV1, TRPV2, SP, CGRP and VGLUT2. Perivascular nerve fibers also had TRPV1, TRPV2, SP, CGRP, VIP, NPY and TH. However, PV- and SPARCL1-containing nerve endings could not be seen in the external auditory canal. It is likely that sensory neurons in the human JG can transduce nociceptive and mechanoreceptive information from the external auditory canal. Theses neurons may be also associated with neurogenic inflammation in the external auditory canal and ear-cough reflex through the vagus nerve.


Decreasing Unplanned Extubations in the Neonatal ICU.
Unplanned extubation (UE) is a preventable adverse event and may lead to additional complications such as cardiovascular resuscitation or respiratory compromise in a critically ill neonate during an emergent re-intubation. A quality improvement project to reduce unplanned endotracheal tube dislodgement would reduce these morbidities. We aimed to reduce UEs in the NICU to 1 UE/100 ventilator days by October 2018. As of the baseline period (March 2017 to November 2017), our level 4 NICU had 950 annual admissions and a baseline rate of 9.9 UEs/100 ventilator days. We formed an inter-professional task force consisting of a neonatologist, 2 respiratory therapists, and the NICU nurse educator. We tracked all of our UE events and required the staff involved to file an electronic safety report. PDSA (plan-do-study-act) cycles consisted of staff attitude survey, development of a data collection tool, protocol of 2 staff members for all transfers of intubated patients, staff education around securement device, and daily retaping of endotracheal tubes to securement device. UE events and ventilator days were extracted from a respiratory database and the electornic medical record. A special cause variation was noted via control chart rules for the mean UE rate from a baseline of 9.9 UEs/100 ventilator days in the baseline period compared to a post-intervention mean of 1.6 UEs/100 ventilator days for the period of August 2018 to March 2019). During the intervention phase of the project (December 2017 to July 2018), a special cause variation was noted with a UE rate of 5/100 ventilator days. Development of a quality improvement project by a multidisciplinary taskforce, along with several PDSA cycles including education and staff awareness, reduced the UE rate by 84% in a level 4 NICU. Ongoing surveillance, education, and review of UE cases will be key to maintaining UE events at a goal of 1 UE/100 ventilator days.


Telencephalic enkephalinergic systems in the rat brain.
The comparative distribution of Leu-enkephalin and an enkephalin precursor fragment, BAM-22P, was studied in the rat telencephalon using immunocytochemical procedures. Although the enkephalins have previously been localized to telencephalic neurons, the use of high doses of colchicine in the present study has permitted the visualization of widespread enkephalin-containing systems in cortical-limbic regions. Rats were treated intraventricularly with colchicine (300 to 400 micrograms) to enhance the visualization of neuronal perikarya. Leu-enkephalin immunoreactivity was noted in perikarya as well as processes of neurons in many telencephalic regions including the olfactory bulb, parolfactory areas, olfactory tubercle, basal ganglia, nucleus accumbens, septal nuclei, amygdala, hippocampal formation, and several paleocortical regions such as the piriform, entorhinal, and cingulate cortex, as well as regions of the frontal, parietal, and occipital neocortex. In the majority of regions investigated, analysis of sections adjacent to those processed for Leu-enkephalin histochemistry showed BAM-22P immunoreactivity to be localized to perikarya and processes in the same region. These results suggest that brain enkephalin-positive neurons may possess mechanisms for enkephalin biosynthesis similar to those found in adrenal medullary enkephalin cells.


Prevention of marine biofouling using natural compounds from marine organisms.
All surfaces that are submerged in the sea rapidly become covered by a biofilm. This process, called biofouling, has substantial economic consequences. Paints containing tri-butyl-tin (TBT) and copper compounds are used to protect marine structures by reducing biofouling. However, these compounds have damaging effects on the marine environment, as they are not biodegradable. It has been noted that many seaweeds and invertebrates found in the sea are not covered by a mature biofilm. This is due to the release of compounds into the surrounding seawater that deter the settlement of fouling organisms. In addition, seaweeds and invertebrates have bacteria on their surfaces that produce compounds to deter settling organisms. The production of compounds by bacteria and their living hosts work in concert to protect the hosts' surfaces. All of these compounds can be collected so they may be natural alternatives to TBT and copper compounds. However, the benefits associated with the use of bacteria as sources of these compounds means that bacteria are the organisms of choice for obtaining natural products for antifouling coatings.


[Significance of evaluation of the functional status of the vessels and the heart after surgical treatment of injuries of the cervical and thoracic levels of the spinal cord].
An analysis of the clinical and neurological data on 52 patients with spinal cord lesions at the level of C4-Th8 segments and their comparison with the function of the vessels and heart have shown that the severity of these patients' condition is determined by disorders of the regulation of the vascular tone, the development of arteriolar hypotonia, disturbances of the regulation of cardiac activity and pulmonary ventilation disorders. The sympathetic regulation of the vessels and heart undergoes phasic changes which should be taken into consideration in the anesthetic-resuscitative support of surgical intervention. This helps decrease the mortality rate and improve the results of treatment.


Community structure and abundance of ammonia-oxidizing archaea and bacteria after conversion from soybean to rice paddy in albic soils of Northeast China.
Community composition of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) in the albic soil grown with soybean and rice for different years was investigated by construction of clone libraries, denaturing gradient gel electrophoresis (DGGE), and quantitative polymerase chain reaction (q-PCR) by PCR amplification of the ammonia monooxygenase subunit A (amoA) gene. Soil samples were collected at two layers (0-5 and 20-25 cm) from a soybean field and four rice paddy fields with 1, 5, 9, and 17 years of continuous rice cultivation. Both the community structures and abundances of AOA and AOB showed detectable changes after conversion from soybean to rice paddy judged by clone library, DGGE, and q-PCR analyses. In general, the archaeal amoA gene abundance increased after conversion to rice cultivation, while bacterial amoA gene abundance decreased. The abundances of both AOA and AOB were higher in the surface layer than the bottom one in the soybean field, but a reverse trend was observed for AOB in all paddy samples regardless of the duration of paddy cultivation. Phylogenetic analysis identified nine subclusters of AOA and seven subclusters of AOB. Community composition of both AOA and AOB was correlated with available ammonium and increased pH value caused by flooding in multiple variance analysis. Community shift of AOB was also observed in different paddy fields, but the two layers did not show any detectable changes in DGGE analysis. Conversion from soybean to rice cultivation changed the community structure and abundance of AOA and AOB in albic agricultural soil, which requires that necessary cultivation practice be followed to manage the N utilization more effectively.


Prediction of T staging in PI-RADS 4-5 prostate cancer by combination of multiparametric MRI and 68Ga-PSMA-11 PET/CT.
In this study, we explored the diagnostic performances of multiparametric magnetic resonance imaging (mpMRI), 68 Ga-PSMA-11 PET/CT and combination of 68 Ga-PSMA-11 PET/CT and mpMRI (mpMRI + PET/CT) for extracapsular extension (ECE). Based on the analyses above, we tested the feasibility of using mpMRI + PET/CT results to predict T staging in prostate cancer patients. By enrolling 75 patients of prostate cancer with mpMRI and 68 Ga-PSMA-11 PET/CT before radical prostatectomy, we analyzed the detection performances of ECE in mpMRI, 68 Ga-PSMA-11 PET/CT and mpMRI + PET/CT on their lesion images matched with their pathological sample images layer by layer through receiver operating characteristics (ROC) analysis. By inputting the lesion data into Prostate Imaging Reporting and Data System (PI-RADS), we divided the lesions into different PI-RADS scores. The improvement of detecting ECE was analyzed by net reclassification improvement (NRI). The predictors for T staging were evaluated by using univariate and multivariable analysis. The Kappa test was used to evaluate the prediction ability. One hundred three regions of lesion were identified from 75 patients. 50 of 103 regions were positive for ECE. The ECE diagnosis AUC of mpMRI + PET/CT is higher than that of mpMRI alone (ΔAUC = 0.101; 95% CI, 0.0148 to 0.1860; p < 0.05, respectively). Compared to mpMRI, mpMRI + PET/CT has a significant improvement in detecting ECE in PI-RADS 4-5 (NRI 36.1%, p < 0.01). The diagnosis power of mpMRI + PET/CT was an independent predictor for T staging (p < 0.001) in logistic regression analysis. In patients with PI-RADS 4-5 lesions, 40 of 46 (87.0%) patients have correct T staging prediction from mpMRI + PET/CT (κ 0.70, p < 0.01). The prediction of T staging in PI-RADS 4-5 prostate cancer patients by mpMRI + PET/CT had a quite good performance.


Intraoperative pulseless electrical activity and acute cardiogenic shock after administration of phenylephrine, epinephrine, and ketamine.
The use of phenylephrine has been well described as a potential cause of morbidity and mortality. A thorough literature review of phenylephrine use is presented in this article. The use of ketamine and epinephrine with phenylephrine can precipitate an even more potentially lethal and catastrophic syndrome. We present the case of a 21-year-old man with Hodgkin's lymphoma and lupus who experienced an abrupt hypertensive crisis followed by pulseless electrical activity and cardiogenic shock after application of 2.5% phenylephrine-soaked nasal pledgets prior to excision of a large nasopharyngeal tumor. This case report adds to the current literature on the potential dangers of phenylephrine in clinical practice and describes a case of reversible severe left ventricular dysfunction in the setting of excessive pharmacologically induced sympathetic stimulation.


Analyzing incomplete longitudinal clinical trial data.
Using standard missing data taxonomy, due to Rubin and co-workers, and simple algebraic derivations, it is argued that some simple but commonly used methods to handle incomplete longitudinal clinical trial data, such as complete case analyses and methods based on last observation carried forward, require restrictive assumptions and stand on a weaker theoretical foundation than likelihood-based methods developed under the missing at random (MAR) framework. Given the availability of flexible software for analyzing longitudinal sequences of unequal length, implementation of likelihood-based MAR analyses is not limited by computational considerations. While such analyses are valid under the comparatively weak assumption of MAR, the possibility of data missing not at random (MNAR) is difficult to rule out. It is argued, however, that MNAR analyses are, themselves, surrounded with problems and therefore, rather than ignoring MNAR analyses altogether or blindly shifting to them, their optimal place is within sensitivity analysis. The concepts developed here are illustrated using data from three clinical trials, where it is shown that the analysis method may have an impact on the conclusions of the study.


Novel treatments after experimental brain injury.
Perinatal hypoxic-ischaemic encephalopathy(HIE) is being studied in laboratory models that allow the delayed cascade of events triggered by the energetic insult to be examined in detail. The concept of the 'excitotoxic cascade' provides a conceptual framework for thinking about the pathogenesis of HIE. Major events in the cascade triggered by hypoxia-ischaemia include overstimulation of N-methyl-D-aspartate type glutamate receptors, calcium entry into cells, activation of calcium-sensitive enzymes such as nitric oxide synthase, production of oxygen free radicals, injury to mitochondria, leading in turn to necrosis or apoptosis. New experimental approaches to salvaging brain tissue from the effects of HIE include inhibition of neuronal nitric oxide synthase, administration of neuronal growth factors, and inhibition of the caspase enzymes that execute apoptosis. Recent experimental work suggests that these approaches may be effective during a longer 'therapeutic window' after the insult, because they are acting on events that are relatively delayed. Application of modest hypothermia may allow these agents to be neuroprotective at even longer intervals after hypoxia-ischaemia.


Serum-free and xenobiotic-free preservation of cultured human limbal epithelial cells.
To develop a one-week storage method, without serum and xenobiotics, that would maintain cell viability, morphology, and phenotype of cultured human limbal epithelial sheets. Human limbal explants were cultured on intact human amniotic membranes for two weeks. The sheets were stored in a hermetically sealed container at 23°C in either a serum-free medium with selected animal serum-derived compounds (Quantum 286) or a xenobiotic-free medium (Minimal Essential Medium) for 4 and 7 days. Stored and non-stored cultures were analyzed for cell viability, amniotic membrane and epithelial sheet thickness, and a panel of immunohistochemical markers for immature cells (ΔNp63α, p63, Bmi-1, C/EBP∂, ABCG2 and K19), differentiated cells (K3 and Cx43), proliferation (PCNA), and apoptosis (Caspase-3). The cell viability of the cultures was 98 ± 1% and remained high after storage. Mean central thickness of non-stored limbal epithelial sheets was 23 ± 3 μm, and no substantial loss of cells was observed after storage. The non-stored epithelial sheets expressed a predominantly immature phenotype with ΔNp63α positivity of more than 3% in 9 of 13 cultures. After storage, the expression of ABCG2 and C/EBP∂ was reduced for the 7 day Quantum 286-storage group; (P = 0.04), and Bmi-1 was reduced after 4 day Quantum 286-storage; (P = 0.02). No other markers varied significantly. The expression of differentiation markers was unrelated to the thickness of the epithelia and amniotic membrane, apart from ABCG2, which correlated negatively with thickness of limbal epithelia (R = -0.69, P = 0.01) and ΔNp63α, which correlated negatively with amniotic membrane thickness (R = -0.59, P = 0.03). Limbal epithelial cells cultured from explants on amniotic membrane can be stored at 23°C in both serum-free and xenobiotic-free media, with sustained cell viability, ultrastructure, and ΔNp63α-positivity after both 4 and 7 days.


The tdh and serA operons of Escherichia coli: mutational analysis of the regulatory elements of leucine-responsive genes.
The tdh promoter of Escherichia coli is induced seven- to eightfold when cells are grown in the presence of exogenous leucine. A scheme was devised to select mutants that exhibited high constitutive expression of the tdh promoter. The mutations in these strains were shown to lie within a previously identified gene (lrp) that encodes Lrp (leucine-responsive regulatory protein). By deletion analysis, the site of action of Lrp was localized to a 25-bp region between coordinates -69 and -44 of the tdh promoter. Disruption of a 12-bp presumptive target sequence found in this region of tdh resulted in constitutively derepressed expression from the tdh promoter. Similar DNA segments (consensus, TTTATTCtNaAT) were also identified in a number of other promoters, including each of the Lrp-regulated promoters whose nucleotide sequence is known. The sequence of the promoter region of serA, an Lrp-regulated gene, was determined. No Lrp consensus target sequence was present upstream of serA, suggesting that Lrp acts indirectly on the serA promoter. A previously described mutation in a leucine-responsive trans-acting factor, LivR (J. J. Anderson, S. C. Quay, and D. L. Oxender, J. Bacteriol. 126:80-90, 1976), resulted in constitutively repressed expression from the tdh promoter and constitutively induced expression from the serA promoter. The possibility that LivR and Lrp are allelic is discussed.


rAAV2/7 vector-mediated overexpression of alpha-synuclein in mouse substantia nigra induces protein aggregation and progressive dose-dependent neurodegeneration.
Alpha-synuclein is a key protein implicated in the pathogenesis of Parkinson's disease (PD). It is the main component of the Lewy bodies, a cardinal neuropathological feature in the disease. In addition, whole locus multiplications and point mutations in the gene coding for alpha-synuclein lead to autosomal dominant monogenic PD. Over the past decade, research on PD has impelled the development of new animal models based on alpha-synuclein. In this context, transgenic mouse lines have failed to reproduce several hallmarks of PD, especially the strong and progressive dopaminergic neurodegeneration over time that occurs in the patients. In contrast, viral vector-based models in rats and non-human primates display prominent, although highly variable, nigral dopaminergic neuron loss. However, the few studies available on viral vector-mediated overexpression of alpha-synuclein in mice report a weak neurodegenerative process and no clear Lewy body-like pathology. To address this issue, we performed a comprehensive comparative study of alpha-synuclein overexpression by means of recombinant adeno-associated viral vectors serotype 2/7 (rAAV2/7) at different doses in adult mouse substantia nigra. We noted a significant and dose-dependent alpha-synucleinopathy over time upon nigral viral vector-mediated alpha-synuclein overexpression. We obtained a strong, progressive and dose-dependent loss of dopaminergic neurons in the substantia nigra, reaching a maximum of 82% after 8 weeks. This effect correlated with a reduction in tyrosine hydroxylase immunoreactivity in the striatum. Moreover, behavioural analysis revealed significant motor impairments from 12 weeks after injection on. In addition, we detected the presence of alpha-synuclein-positive aggregates in the remaining surviving neurons. When comparing wild-type to mutant A53T alpha-synuclein at the same vector dose, both induced a similar degree of cell death. These data were supported by a biochemical analysis that showed a net increase in soluble and insoluble alpha-synuclein expression over time to the same extent for both alpha-synuclein variants. In conclusion, our in vivo data provide evidence that strong and significant alpha-synuclein-induced neuropathology and progressive dopaminergic neurodegeneration can be achieved in mouse brain by means of rAAV2/7.


A (99m)Tc-labeled dual-domain cytokine ligand for imaging of inflammation.
Interleukin (IL)-1 and IL-18 are potent proinflammatory cytokines in inflammation-related diseases. Their actions are regulated by IL-1 receptor antagonist (IL-1ra) and IL-18 binding protein (IL-18bp). This study was designed to (99m)Tc-radiolabel an IL-1ra and IL-18bp dual-domain cytokine ligand, IL-18bp-Fc-IL-1ra, for specific inflammation targeting. The (99m)Tc-IL-18bp-Fc-IL-1ra was obtained by direct labeling via 2-iminothiolane reduction. Competitive binding of (99m)Tc-labeled and unlabeled IL-18bp-Fc-IL-1ra to rat polymorphonuclear leukocytes was assessed in vitro. A mouse ear edema model was used to evaluate specific targeting properties of (99m)Tc-IL-18bp-Fc-IL1ra in vivo. The correlation between (99m)Tc-IL-18bp-Fc-IL-1ra uptake and (111)In-labeled polymorphonuclear neutrophil infiltration was studied using ischemic-reperfused rat hearts. Direct (99m)Tc-labeling yielded a stable dual-domain cytokine radioligand with radiochemical purity greater than 95% after gel filtration. Competitive binding studies showed specific targeting of (99m)Tc-IL-18bp-Fc-IL-1ra to inflammatory cells. The (99m)Tc-IL-18bp-Fc-IL-1ra uptake was 1.80±0.17 % injected dose per gram (%ID/g) in the inflamed ear without blocking, whereas uptake in the presence of IL-18bp-Fc-IL-1ra was 1.09±0.08 %ID/g (P<.05). The amounts of IL-1β and IL-18 were significantly increased in the inflamed ears compared to the vehicle controls. A significant correlation of (99m)Tc-IL-18bp-Fc-IL-1ra with (111)In-labeled neutrophil distribution was observed in the ischemic-reperfused hearts (P<.001). Targeting proinflammatory cytokines with (99m)Tc-IL-18bp-Fc-IL-1ra may provide a suitable approach for specific detection of inflammatory sites.


Human basophils secrete IL-3: evidence of autocrine priming for phenotypic and functional responses in allergic disease.
Although IL-3 is commonly recognized for its growth factor-like activity, in vitro studies have long demonstrated a unique capacity for this cytokine to also augment the proinflammatory properties and phenotype of human basophils. In particular, basophils secrete mediators that are hallmarks in allergic disease, including vasoactive amines (e.g., histamine), lipid metabolites (e.g., leukotriene C(4)), and cytokines (e.g., IL-4/IL-13), which are all markedly enhanced with IL-3 pretreatment. This priming phenomenon is observed in response to both IgE-dependent and IgE-independent stimulation. Additionally, IL-3 directly activates basophils for IL-13 secretion and enhanced CD69 expression, two markers that are elevated in allergic subjects. Lymphocytes are commonly thought to be the source of the IL-3 that primes for these basophil responses. However, we demonstrate herein for the first time that basophils themselves rapidly produce IL-3 (within 4 h) in response to IgE-dependent activation. More importantly, our findings definitively show that basophils rapidly bind and utilize the IL-3 they produce, as evidenced by functional and phenotypic activity that is inhibited in the presence of neutralizing anti-IL-3 receptor (CD123) Abs. We predict that autocrine IL-3 activity resulting from low-level IgE/FcepsilonRI cross-linking by specific allergen represents an important mechanism behind the hyperreactive nature of basophils that has long been observed in allergic disease.


Racial pride and religiosity among African American boys: implications for academic motivation and achievement.
The persistent underachievement among African American boys has led to increased empirical inquiry, yet little research considers within-group variation in achievement nor positive youth characteristics that help explain positive achievement outcomes. This study conceptualized culturally-based factors (racial pride and religiosity) as adolescent assets that would promote African American boys' achievement and also enhance positive effects of other youth assets (positive educational utility beliefs) on achievement. Our sample included 158 adolescent boys (M = 17.08) from a large, socioeconomically diverse suburban community context. Accounting for demographic background variables, educational utility beliefs were positively associated with academic grade performance. A significant educational utility beliefs and racial pride interaction indicated a stronger, positive association of educational utility beliefs with grade performance among boys with higher racial pride relative to those with lower racial pride. Also, there was a stronger positive association between educational utility beliefs and grades for boys reporting lower religious importance, but boys endorsing both lower educational utility beliefs and religious importance were at highest risk for low grade performance. Overall results suggest the importance of considering culturally-based factors in studying achievement motivation processes among ethnic minority adolescents.


Rheumatoid arthritis in Jordan: a cross sectional study of disease severity and associated comorbidities.
Treating rheumatoid arthritis (RA) to target is advocated using disease activity measures. The impact of RA on the general health status of affected patients in Jordan is not well described. This study reported the severity of RA in Jordan and its association with consequent disabilities and comorbidities. A cross-sectional, observational study was conducted at King Abdullah University Hospital in the north of Jordan. All patients who were diagnosed with RA were included. Patients' demographics, comorbidities, disease activity score (DAS 28), and clinical disease activity index (CDAI) were collected. Both DAS 28 and CDAI were utilized to categorize RA disease activity. A total of 465 patients with RA were included: 82% were females; mean age ± standard deviation (SD) was 47.62±14.6 years; and mean disease duration ± SD was 6±4.45 years. The mean ± SD for the DAS 28 and CDAI was 5.1±1.5 and 23±14.2, respectively. According to the DAS 28, 51% of the patients were in the high disease activity category and only 5% were in remission. On the other hand, according to the CDAI, 44% were in the high disease activity category and only 1% were in remission. In Jordan, patients with RA have a high severe disease rate and a low remission rate. The disease is often progressive and associated with comorbidities that need to be managed.