Short communication: Association of foot and leg conformation and body weight with claw disorders in Spanish Holstein cows.
Data of first-lactation Holstein cows including claw disorders, foot and leg type traits, and structure and capacity type traits were used to study the phenotypic association of conformation and body weight with claw health status. The claw disorders studied were digital and interdigital dermatitis, sole ulcer, white line disease, and an overall claw disorder trait, indicating the presence of any of the 6 claw disorders recorded by the Spanish I-SAP program. Body weight was estimated indirectly with the Von Bertalanffy equation using live weight, which was also estimated from body depth, stature, and chest width. Cows with poor scores for foot and leg traits were more likely to have a claw disorder, with the exception of rear leg rear view and foot and leg composite that did not show any association with the studied disorders. The heavier the cow was, the higher was the probability of having sole ulcer, white line disease, or overall claw disorder trait, but digital and interdigital dermatitis, as an infectious disorder, did not show any association with body weight. Therefore, it is recommended that the preventive trimming routine be improved, which means checking each cow at least once per lactation and trimming if necessary, to achieve balanced weight-bearing for heavier cows and cows with poor feet and leg classification.


The Postfusion Structure of the Heartland Virus Gc Glycoprotein Supports Taxonomic Separation of the Bunyaviral Families Phenuiviridae and Hantaviridae.
Heartland virus (HRTV) is an emerging human pathogen that belongs to the newly defined family Phenuiviridae, order Bunyavirales Gn and Gc are two viral surface glycoproteins encoded by the M segment and are required for early events during infection. HRTV delivers its genome into the cytoplasm by fusion of the viral envelope and endosomal membranes under low-pH conditions. Here, we describe the crystal structure of HRTV Gc in its postfusion conformation. The structure shows that Gc displays a typical class II fusion protein conformation, and the overall structure is identical to severe fever with thrombocytopenia syndrome virus (SFTSV) Gc, which also belongs to the Phenuiviridae family. However, our structural analysis indicates that the hantavirus Gc presents distinct features in the aspects of subdomain orientation, N-linked glycosylation, the interaction pattern between protomers, and the fusion loop conformation. This suggests their family-specific subunit arrangement during the fusogenic process and supports the recent taxonomic revision of bunyaviruses. Our results provide insights into the comprehensive comparison of class II membrane fusion proteins in two bunyavirus families, yielding valuable information for treatments against these human pathogens.IMPORTANCE HRTV is an insect-borne virus found in America that can infect humans. It belongs to the newly defined family Phenuiviridae, order Bunyavirales HRTV contains three single-stranded RNA segments (L, M, and S). The M segment of the virus encodes a polyprotein precursor that is cleaved into two glycoproteins, Gn and Gc. Gc is a fusion protein facilitating virus entry into host cells. Here, we report the crystal structure of the HRTV Gc protein. The structure displays a typical class II fusion protein conformation. Comparison of HRTV Gc with a recently solved structure of another bunyavirus Gc revealed that these Gc structures display a newly defined family specificity, supporting the recent International Committee on Taxonomy of Viruses reclassification of the bunyaviruses. Our results expand the knowledge of bunyavirus fusion proteins and help us to understand bunyavirus characterizations. This study provides useful information to improve protection against and therapies for bunyavirus infections.


Diet and caries experience among preschool children in Ajman, United Arab Emirates.
The study investigated the association between food and drink consumption and the caries experience among young children in Ajman, United Arab Emirates. A one-stage cluster sample was used to select children who were 5 or 6 yr of age. Clinical examinations for caries were conducted. Parents completed questionnaires seeking information on dietary habits. Principal components analysis was used to derive a summary score for the dietary variables (designated 'snack consumption level') and thus overcome the multicollinearity problems associated with using multiple dietary variables. Dental examination and questionnaire data were obtained for 1,036 children (79.9% participation rate). The overall mean number of decayed, missing or filled teeth (dmft) was 4.5. Snacking three or more times per day was associated with a dmft score that was almost one-third higher than the dmft score for children who snacked only once daily. One-third of children had a low overall snack consumption level (41% moderate and 25% high). There was a consistent dmft gradient across those categories. The severity of Early Childhood Caries (ECC) in young Ajman children is high, with dietary habits being important determinants. Cariogenic snack consumption can be represented using a summary exposure variable that appears to be valid. Young children in Ajman would benefit from health-promotion strategies directed towards appropriate dietary practices.


Inhibition of the thiol-specific antioxidant activity of rat liver MSP23 protein by hemin.
Mouse stress-inducible 23-kDa protein (MSP23) belongs to a new type of antioxidative protein family and is highly expressed in livers. To examine its physiological role in the tissue, we have purified native MSP23 protein from the cytosolic fraction of rat liver. The purified protein showed the thiol-specific antioxidant activity and protected glutamine synthetase from inactivation by a mixed metal-thiol oxidation. We examined the effect of hemin on that activity since it was recently shown by others that the rat liver MSP23 protein had a high binding affinity to heme. We have found that hemin at low concentrations inhibits the antioxidative activity of MSP23. This result suggests that one type of oxidant damage caused by hemin to cells is due to inactivation of the antioxidative protein, MSP23.


Treatment of haemothorax.
Haemothorax is a problem commonly encountered in medical practice and is most frequently related to open or closed chest trauma or to invasive procedures of the chest. Spontaneous haemothorax is less common and can have various causes, such as the use of anticoagulants, neoplasia, and rupture of pleural adhesions. Identification by radiography and thoracentesis is indicated and treatment of the underlying trauma should start immediately. After insertion of a large chest tube, antibiotic prophylaxis in trauma patients should be administered for 24 h. Further treatment depends on the haemodynamic stability of the patient, the volume of evacuated blood and the occurrence of persistent blood loss. Surgical exploration by VATS or thoracotomy is necessary if >1.500 ml of blood has accumulated and/or an ongoing production of >200 ml of blood per hour is observed. If the haemorrhage is less severe, careful investigation into the underlying cause must be performed and blood should be evacuated by tube thoracostomy. If clotted blood retained in spite of tube thoracostomy, intrapleural fibrinolytic therapy can be applied to breakdown clots and adhesions. If conservative treatment is insufficient, a surgical approach with VATS or thoracotomy is indicated to prevent subsequent complications.


Energy Harvesting from Drops Impacting onto Charged Surfaces.
We use a combination of high-speed video imaging and electrical measurements to study the direct conversion of the impact energy of water drops falling onto an electrically precharged solid surface into electrical energy. Systematic experiments at variable impact conditions (initial height; impact location relative to electrodes) and electrical parameters (surface charge density; external circuit resistance; fluid conductivity) allow us to describe the electrical response quantitatively without any fit parameters based on the evolution of the drop-substrate interfacial area. We derive a scaling law for the energy harvested by such "nanogenerators" and find that optimum efficiency is achieved by matching the timescales of the external electrical energy harvesting circuit and the hydrodynamic spreading process.


Iron-Facilitated Transformation of Mesoporous Spinel Nanosheets into Oxyhydroxide Active Species in the Oxygen Evolution Reaction.
The oxygen evolution reaction (OER) is critical for many clean energy conversion and storage technologies because it contributes the electrons required for converting renewable electricity into value-added chemicals. Electrocatalysts can promote the sluggish oxygen evolution process involving four-electron transfer. Herein, we prepare mesoporous spinel oxide nanosheets and develop an efficient strategy using Fe substitution to enable mesoporous NiCo2O4 nanosheets to generate superior active centers for the OER. Additionally, the iron substitution also promotes the preoxidation of Co/Ni and facilitates the formation of active species. Raman spectroscopy data reveal that the active species of mesoporous NiCo2O4 nanosheets for the OER is NiCo2O4 itself, and the active species of Fe substitution in NiCo2O4 nanosheets are Ni(Co) oxyhydroxides. Therefore, the iron substitution is beneficial to facilitate the transformation of spinel NiCo2O4 into active Ni(Co) oxyhydroxides under OER conditions. Owing to the mesoporous nanosheet structure and the formation of oxyhydroxide active species, the optimized mesoporous Fe0.2Ni0.8Co2O4 nanosheet catalyst exhibits a low overpotential of 270 mV to deliver a current density of 10 mA cm-2 and a small Tafel slope of 39 mV dec-1 for the oxygen evolution reaction in alkaline media.


The propositional nature of human associative learning.
The past 50 years have seen an accumulation of evidence suggesting that associative learning depends on high-level cognitive processes that give rise to propositional knowledge. Yet, many learning theorists maintain a belief in a learning mechanism in which links between mental representations are formed automatically. We characterize and highlight the differences between the propositional and link approaches, and review the relevant empirical evidence. We conclude that learning is the consequence of propositional reasoning processes that cooperate with the unconscious processes involved in memory retrieval and perception. We argue that this new conceptual framework allows many of the important recent advances in associative learning research to be retained, but recast in a model that provides a firmer foundation for both immediate application and future research.


Pegylated insulin-like growth factor-1 biotherapeutic delivery promotes rotator cuff regeneration in a rat model.
Tears in the rotator cuff are challenging to repair because of the complex, hypocellular, hypovascular, and movement-active nature of the tendon and its enthesis. Insulin-like Growth Factor-1 (IGF-1) is a promising therapeutic for this repair. However, its unstable nature, short half-life, and ability to disrupt homeostasis has limited its clinical translation. Pegylation has been shown to improve the stability and sustain IGF-1 levels in the systemic circulation without disrupting homeostasis. To provide localized delivery of IGF-1 in the repaired tendons, we encapsulated pegylated IGF-1 mimic and its controls (unpegylated IGF-1 mimic and recombinant human IGF-1) in polycaprolactone-based matrices and evaluated them in a pre-clinical rodent model of rotator cuff repair. Pegylated-IGF-1 mimic delivery reestablished the characteristic tendon-to-bone enthesis structure and improved tendon tensile properties within 8 weeks of repair compared to controls, signifying the importance of pegylation in this complex tissue regeneration. These results demonstrate a simple and scalable biologic delivery technology alternative to tissue-derived grafts for soft tissue repair.


Withholding of population funds, 13 August 1987.
The United States Agency for International Development (USAID) withheld from the United Nations Population Fund (UNFPA) $25 million appropriated for the Fund by the United States Congress. This was the third year that funds had been withheld. As in earlier years, the Reagan administration objected to UNFPA support for China. It viewed China's family planning program, which emphasized the importance of limiting family size to one child, as sanctioning coercive abortions.


The effectiveness of computer-managed instruction versus traditional classroom lecture on achievement outcomes.
This controlled experimental study examines the effect of two teaching methods on achievement outcomes from a 15-week, 2 credit hour semester course taught at two midwestern universities. Students were randomly assigned to either computer-managed instruction in which faculty function as tutors or the traditional classroom course of study. In addition, the effects of age, grade point average, attitudes toward computers, and satisfaction with the course on teaching method were analyzed using analysis of covariance. Younger students achieved better scores than did older students. Regardless of teaching method, however, neither method appeared to be better than the other for teaching course content. Students did not prefer one method over the other as indicated by their satisfaction scores. With demands upon university faculty to conduct research and publish, alternative methods of teaching that free faculty from the classroom should be considered. This study suggests that educators can select such an alternative teaching method to traditional classroom teaching without sacrificing quality education for certain courses.


Photosensitization of murine tumor, vasculature and skin by 5-aminolevulinic acid-induced porphyrin.
The effects of topical and systemic administration of 5-aminolevulinic acid (ALA) were examined in several murine tumor systems with regard to porphyrin accumulation kinetics in tumor, skin and blood, vascular and tumor cell photosensitization and tumor response after light exposure. Marked, transient increases in porphyrin levels were observed in tumor and skin after systemic and topical ALA. Rapid, transient, dose-dependent porphyrin increases were also observed in blood; these were pronounced after systemic ALA injection and mild after topical application. They were highest within 1 h after ALA injection, thereafter declining rapidly. This matched the clearing kinetics of injected exogenous protoporphyrin IX (PpIX). Initially, vascular photosensitivity changed inversely to blood porphyrin levels, increasing gradually up to 5 h post-ALA, as porphyrin was clearing from the bloodstream. This pattern was again matched by injected, exogenous PpIX. After therapeutic tumor treatment vascular disruption of the tumor bed, while observed, was incomplete, especially at the tumor base. Minimal direct tumor cell kill was found at low photodynamic therapy (PDT) doses (250 mg/kg ALA, 135 J/cm2 light). Significant, but limited (< 1 log) direct photodynamic tumor cell kill was obtained when the PDT dose was raised to 500 mg/kg systemic ALA, followed 3 h later by 270 J/cm2, a dose that was however toxic to the animals. The further reduction of clonogenic tumor cells over 24 h following treatment was moderate and probably limited by the incomplete disruption of the vasculature. Tumor responses were highest when light treatment was carried out at the time of highest tumor porphyrin content rather than at the time of highest vascular photosensitivity. Tumor destruction did not reach the tumor base, regardless of treatment conditions.


Access to data: comparing AccessMed with Query by Review.
To evaluate the performance of tools for authoring patient database queries. Query by Review, a tool that exploits the training that users have undergone to master a result review system, was compared with AccessMed, a vocabulary browser that supports lexical matching and the traversal of hierarchical and semantic links. Seven subjects (Medical Logic Module authors) were asked to use both tools to gather the vocabulary terms necessary to perform each of eight laboratory queries. The proportion of queries that were correct; intersubject agreement. Query by Review had better performance than AccessMed (38% correct queries versus 18%, p = 0.002), but both figures were low. Poor intersubject agreement (28% for Query by Review and 21% for AccessMed) corroborated the relatively low performance. Subjects appeared to have trouble distinguishing laboratory tests from laboratory batteries, picking terms relevant to the particular data type required, and using classes in the vocabulary's hierarchy. Query by Review, with its more constrained user interface, performed somewhat better than AccessMed, a more general tool. Neither tool achieved adequate performance, however, which points to the difficulty of formulating a query for a clinical database and the need for further work.


An RCT of an Online Parenting Program for Parents of Preschool-Aged Children With ADHD Symptoms.
Objective: This randomized control trial evaluated the efficacy of an online self-help program in a sample of parents of preschoolers with ADHD symptoms. Method: Parents were randomly assigned to the intervention group (n = 27) or the delayed intervention group (n = 26). Child behavior measures were completed by mothers, fathers, and teachers, and parenting measures were completed by mothers. Results: Intent-to-treat analyses indicated significant post-intervention improvements in mother-rated child hyperactivity/inattention, restlessness/impulsivity, defiance/aggression, social functioning, and teacher-rated prosocial behavior, as well as significant improvements in maternal over-reactivity, verbosity, laxness, positive parenting, parenting satisfaction, self-efficacy, stress, and depression. At 6-month follow-up, effects were maintained for maternal over-reactivity and verbosity, parenting satisfaction and self-efficacy, and parental stress and depression. Conclusion: This study provides evidence for the effectiveness of an online self-help parenting program in reducing preschool inattentive behavior difficulties, and in increasing parenting competence, satisfaction in the parenting role, and maternal well-being.


Inventory of real-world data sources in Japan: Annual survey conducted by the Japanese Society for Pharmacoepidemiology Task Force.
The Database Task Force of the Japan Society for Pharmacoepidemiology began its annual surveys of databases available for clinico and pharmacoepidemiological studies in 2010. In this report, we summarize the characteristics of the databases available in Japan based on the results of our 2021 survey to illustrate the recent developments in the infrastructure for database research in Japan. We included 20 major databases from the academia, government, or industry that were accessible to third parties. We used a web-based questionnaire to ask the database providers about their characteristics, such as their organization, data source(s), numbers of individuals enrolled, age distribution, code(s) used, and average follow-up periods. We received responses from all 20 databases approached: eight hospital-based databases, six insurer-based databases, four pharmacy-based databases, and two in the "other" category. Among them, 17 contained information from medical claims, pharmacy claims, and/or Diagnosis Procedure Combination data. Most insurer databases contained health check-up data that could be attached to the claims component. Some hospital-based databases had data from electronic medical records. Most insurer-based databases collected data from the insurers of working-age employees and therefore had limited coverage of older people. Most databases coded their medication data using the Japanese reimbursement codes, and many provided Anatomical Therapeutic Chemical Classification codes. The number of databases available for clinico and pharmacoepidemiological research and the proportion of the population they cover are increasing in Japan. The differences in their characteristics mean that the appropriate database must be selected for a particular study purpose.


Anxiety and Alzheimer's disease.
This study investigated symptoms of anxiety in two samples of clinic outpatients diagnosed with Alzheimer's disease (AD). Clinician and caregiver reports were obtained using standardized measures to characterize a broad array of anxiety symptoms. Anxiety symptoms were reported for a substantial proportion of subjects, regardless of whether clinician or caregiver ratings were used. Anxious or worried appearance was most common (68% to 71%), followed by fearfulness, tension, restlessness, and fidgeting (37% to 57%). Sleep disturbance and various somatic symptoms were less common (8% to 34%). Although anxiety symptoms were prevalent, only 5% to 6% of subjects met Diagnostic and Statistical Manual of Mental Disorders criteria for the diagnosis of generalized anxiety disorder. In both samples, anxiety symptoms were associated with depression, behavioral disturbances, and increased cognitive impairment. Study findings support a high occurrence of anxiety in patients with dementia, and treatments for anxiety might therefore be helpful in reducing the psychiatric burden of AD.


Different activation pathways involved in antibody-dependent and immune-complexes-triggered cytotoxicity mediated by neutrophils.
We have shown previously that normal human neutrophils triggered by immune complexes displayed significant levels of cytotoxicity towards non-sensitized target cells (non-specific cytotoxicity-NSC) (Geffner, J. R. et al. 1987). Despite the fact that NSC and antibody-dependent cellular cytotoxicity (ADCC) are both mediated through neutrophil Fc gamma R and require the activation of the respiratory burst, the cytolytic mechanisms involved in each case appear to be different. In order to analyse the pathways of activation involved in the induction of NSC and ADCC, we studied here some of the metabolic requirements associated with each cytotoxic function. Our results suggest that ADCC is dependent on Na+/H+ antiporter activity, de novo protein synthesis, availability of external Ca2+ and calmodulin activity, activation of phospholipase C and activation of protein kinase C. On the other hand, NSC appears to be dependent on availability of external Ca2+ and calmodulin activity and activation of phospholipase A2. These results indicate that different pathways of activation are involved in the induction of neutrophil-mediated ADCC and NSC.


Distinctive sequence organization and functional programming of an Alu repeat promoter.
Plasmid clones containing a human Alu family repeat can be transcribed efficiently by RNA polymerase III in HeLa cell extract. This generated three RNA species, all of which initiated from the first base (+1) of the repeat. By studying the transcriptional properties of deletion clones, subclones, and topologically different DNA templates, we demonstrated that: supercoiled DNA templates are transcribed 3- to 5-fold more efficiently than are linear or nicked circular DNA molecules; a contiguous DNA helix in the transcription complexes that extends into the 5' flanking region of positions -30 to -85 is absolutely required for initiation to occur (this interaction does not involve recognition of specific DNA sequences); and similar to the adenovirus VAI RNA and tRNA genes, the Alu repeat 3' to the alpha 1-globin gene (designated 3'-alpha 1 Alu) contains a split intragenic promoter: an anterior element (positions +4 to +37) and a posterior element (positions +70 to +82). However, the promoter of the Alu repeat functions in distinctive ways in comparison to those of other RNA polymerase III-dependent genes. The posterior promoter element alone is sufficient and necessary for an accurate initiation to occur. The presence of the anterior promoter element, which by itself does not initiate transcription, enhances the transcriptional efficiency by a factor of 10- to 20-fold. Furthermore, the distance between the initiation sites and the posterior promoter element, but not the anterior promoter element, remains constant. These results suggest that the promoter of this Alu family repeat consists of at least two functionally different domains: a "directing element" (the posterior promoter element) that determines the accuracy of initiation and an "enhancing element" (the anterior promoter element) that is mainly responsible for the transcriptional efficiency.


Keratoconus and blindness in 469 institutionalised subjects with Down syndrome and other causes of mental retardation.
The records of 469 institutionalised mentally retarded subjects were examined. Keratoconus and blindness were present in 8 per cent and 4 per cent of the ninety-one subjects with Down syndrome. Only one case of keratoconus was found among the 378 subjects with mental deficiency of other aetiology. The incidence of strabismus, nystagmus and cataract was also increased in subjects with Down syndrome, especially in females. Cases of acute keratoconus associated with Down syndrome reported in the world literature are summarised. A total of about forty cases has ben reported between the ages of nine and fifty-three years. Sight has been preserved equally well after both conservative management and keratoplasty in eyes with and without cataracts.


Access to outreach specialist palliative care teams among cancer patients in Denmark.
Equal access to end-of-life care is important. However, social inequality has been found in relation to place-of-death. The question is whether social and economic factors play a role in access to specialist palliative care services. The study analyzed the association between access to outreach specialist palliative care teams (SPCTs) and socioeconomic characteristics of Danish cancer patients who died of their cancer. The study was a population-based, cross-sectional register study. We identified 599 adults who had died of cancer from March 1 to November 30, 2006, in Aarhus County, Denmark. Data from health registers were retrieved and linked based on the unique personal identifier number. Multivariate analysis with adjustment for age, gender, and general practitioner (GP) involvement showed a higher probability of contact with an SPCT among immigrants and descendants of immigrants than among people of Danish origin (prevalence ratio [PR]: 1.55; 95% confidence interval (CI): 1.04;2.31) and among married compared to unmarried patients (PR: 1.25; 95% CI: 1.01;1.54). The trends were most marked among women. We found an association between females, married patients, and female immigrants and their descendants and access to an SPCT in Denmark. However, no association with the examined economic factor was found. Need for specialized health care, which is supposed to be the main reason for access to an SPCT, may be related to economic imbalance; and despite the relative equality found, SPCT access may not be equal for all Danish residents. Further research into social and economic consequences in palliative care services is warranted.


Revealing gene regulation-based neural network computing in bacteria.
Bacteria are known to interpret a range of external molecular signals that are crucial for sensing environmental conditions and adapting their behaviors accordingly. These external signals are processed through a multitude of signaling transduction networks that include the gene regulatory network (GRN). From close observation, the GRN resembles and exhibits structural and functional properties that are similar to artificial neural networks. An in-depth analysis of gene expression dynamics further provides a new viewpoint of characterizing the inherited computing properties underlying the GRN of bacteria despite being non-neuronal organisms. In this study, we introduce a model to quantify the gene-to-gene interaction dynamics that can be embedded in the GRN as weights, converting a GRN to gene regulatory neural network (GRNN). Focusing on Pseudomonas aeruginosa, we extracted the GRNN associated with a well-known virulence factor, pyocyanin production, using an introduced weight extraction technique based on transcriptomic data and proving its computing accuracy using wet-lab experimental data. As part of our analysis, we evaluated the structural changes in the GRNN based on mutagenesis to determine its varying computing behavior. Furthermore, we model the ecosystem-wide cell-cell communications to analyze its impact on computing based on environmental as well as population signals, where we determine the impact on the computing reliability. Subsequently, we establish that the individual GRNNs can be clustered to collectively form computing units with similar behaviors to single-layer perceptrons with varying sigmoidal activation functions spatio-temporally within an ecosystem. We believe that this will lay the groundwork toward molecular machine learning systems that can see artificial intelligence move toward non-silicon devices, or living artificial intelligence, as well as giving us new insights into bacterial natural computing.


Spin blockade, orbital occupation, and charge ordering in La1.5Sr0.5CoO4.
Using Co-L2,3 and O-K x-ray absorption spectroscopy, we reveal that the charge ordering in La1.5Sr0.5CoO4 involves high spin (S=3/2) Co2+ and low spin (S=0) Co3+ ions. This provides evidence for the spin-blockade phenomenon as a source for the extremely insulating nature of the La2-xSrxCoO4 series. The associated e{g}{2} and e{g}{0} orbital occupation accounts for the large contrast in the Co-O bond lengths and, in turn, the high charge ordering temperature. Yet, the low magnetic ordering temperature is naturally explained by the presence of the nonmagnetic (S=0) Co3+ ions. From the identification of the bands we infer that La1.5Sr0.5CoO4 is a narrow band material.


Transversally and axially tunable carbon nanotube resonators in situ fabricated and studied inside a scanning electron microscope.
We report a new design of carbon nanotube (CNT) resonator, whose resonance frequency can be tuned not only transversally by a gate voltage, but also by the axial strain applied through directly pulling the CNT. The resonators are fabricated from individual suspended single-walled CNT (SWCNT) in situ inside a scanning electron microscope. The resonance frequency of a SWCNT resonator can be tuned by more than 20 times with an increase of quality factor when the axial strain of the SWCNT is only increased from nearly zero to 2% at room temperature. The transversal gate-tuning ability is found to be weaker than the axial-tuning ability and decrease with increasing the axial strain. The gate voltage can hardly tune the resonance frequency when the initial axial strain is larger than 0.35% and the CNT acts like a tied string. The relationship among resonance frequency, gate voltage, and initial axial strain of the CNT obtained presently will allow for the designs of CNT resonators with high frequency and large tuning range. The present resonator also shows ultrahigh sensitivity in displacement and force detection, with a resolution being better than 2.4 pm and 0.55 pN, respectively.


The current status of chronic obstructive pulmonary disease awareness, treatments, and plans for improvement in South Korea: a narrative review.
The prevalence of chronic obstructive pulmonary disease (COPD) is high in South Korea which has remained virtually unchanged over the past few years. Other challenges related to COPD in South Korea include a low level of awareness of COPD, underutilization of pulmonary function tests (PFTs) and a low level of inhaler use. Continued efforts have been made to raise awareness of COPD in the general public and promote screening tests for the early detection of COPD patients for high-risk patients, which should be the primary object for better management of the disease. The Korean Academy of Tuberculosis and Respiratory Diseases (KATRD) and the Korea Disease Control and Prevention Agency (KDCA) have been working in tandem on numerous projects to resolve these issues. As the fruit of these efforts, a couple of projects are currently being carried out to add PFTs into the National Health Examination (NHE) of Korea as part of screening tests and improve the quality of COPD treatments in primary care settings. Raising public awareness of a chronic disease such as COPD requires collective efforts of academic societies and government. In addition, personalized education programs that are tailored to individual COPD patients is a necessity to raise adherence of treatment and self-management of COPD. The aim of this manuscript is to report the current status of COPD management in South Korea in hopes that it will help better treat and manage COPD in other nations coping with similar challenges.


A prospective analysis of recreational physical activity and preeclampsia risk.
Although literature suggests that physical activity may reduce preeclampsia risk, most prior studies have relied on retrospective exposure assessment. We aimed to assess prospectively the relation between recreational physical activity before and during pregnancy and risk of preeclampsia. We used data from a 1996-2003 cohort study of 2241 pregnant western Washington State residents. During structured interviews conducted at 15 wk of gestation, on average, women reported recreational activities during the year before pregnancy and during the previous 7 d. Preeclampsia diagnosis was established for 111 women using medical record data and standard guidelines. Recreational activity in the year before pregnancy was nonstatistically significantly associated with reduced preeclampsia risk (adjusted odds ratio [OR] for any vs none 0.55, 95% confidence interval [CI] 0.30-1.02). Any activity during early pregnancy was not strongly associated with preeclampsia risk (adjusted OR = 1.07, 95% CI = 0.67-1.69). Compared with women who reported no recreational activity before or during early pregnancy, risk was nonsignificantly lower among those who were active only before pregnancy (OR = 0.73, 95% CI = 0.30-1.77) and during both periods (OR = 0.76, 95% CI = 0.34-1.73). Those who were active only in early pregnancy had nonsignificantly increased preeclampsia risk (OR = 2.03, 95% CI = 0.71-5.81). These results corroborate previous case-control studies suggesting that prepregnancy recreational activity reduces preeclampsia risk. The absence of an association with early-pregnancy recreational activity in this cohort may be due to misclassification of usual early-pregnancy activity.


Synthesis and Acid-Responsiveness of an Insulated π-Conjugated Polymer Containing Spiropyrans in Its Backbone.
A π-conjugated polymer containing spiropyrans (SPs), which could be almost completely converted to protonated merocyanines (MCH⁺) and back to the SP form by adding an acid and a base, respectively, was developed. The insulation of the π-conjugated polymer, referred to as insulated spiropyran-containing poly(p-phenylene ethynylene) (ins-SP-PPE), using permethylated α-cyclodextrins (PM α-CD) suppressed the π-π interaction between the polymer chains containing MCH⁺, and the installation of PM α-CD improved the switching ability of SPs. The polymer exhibited repeatable acidochromism with almost complete conversion between the SP and MCH⁺ forms. Photoluminescence measurements were conducted and the acid-induced luminescence quenching of the polymer in the solution was observed, which stemmed from energy transfer from the PPE to MCH⁺ moieties. In the solid state, the quantum yield of ins-SP-PPE was more than twice that of the uninsulated polymer, which derived from the insulation effects. The acid-induced luminescence quenching was also observed in the solid state.


Characterization of mineral oxide charging in apolar media.
This paper presents an investigation of the charging behavior of mineral oxide particles dispersed in apolar media. There are a growing number of applications that seek to use electrostatic effects in apolar media to control particle movement and improve aggregation stability. Progress is limited, however, by incomplete knowledge of the mechanism(s) of particle charging in these systems. It has been shown in a number of cases that the acid-base properties of both the particles and the surfactants used to stabilize charge play key roles. A mechanism for acid-base charging has previously been established for mineral oxides in aqueous systems, where the surface hydroxyl groups act as proton donors or receivers depending on the pH of the surrounding solution. In water, the pH at which the surface charge density is zero, i.e., the point of zero charge (PZC), can be used to characterize the acid-base nature of the mineral oxide particles. The current work explores the possible extension of this charging behavior to apolar systems, with the key difference that the surface hydroxyl groups of the mineral oxides react with the surfactant molecules instead of free ions in solution. The apolar charging behavior is explored by measuring the electrophoretic mobility of a series of mineral oxides dispersed in a solution of Isopar-L and AOT, a neutral surfactant in water. The electrophoretic mobility of the particles is found to scale quantitatively, with respect to both sign and magnitude, with their aqueous PZC value. This provides support for the theory of acid-base charging in apolar media and represents a method for predicting and controlling particle charge of mineral oxides dispersed in apolar media.


Mechanisms that underlie μ-opioid receptor agonist-induced constipation: differential involvement of μ-opioid receptor sites and responsible regions.
Reducing the side effects of pain treatment is one of the most important strategies for improving the quality of life of cancer patients. However, little is known about the mechanisms that underlie these side effects, especially constipation induced by opioid receptor agonists; i.e., do they involve naloxonazine-sensitive versus -insensitive sites or central-versus-peripheral μ-opioid receptors? The present study was designed to investigate the mechanisms of μ-opioid receptor agonist-induced constipation (i.e., the inhibition of gastrointestinal transit and colonic expulsion) that are antagonized by the peripherally restricted opioid receptor antagonist naloxone methiodide and naloxonazine in mice. Naloxonazine attenuated the fentanyl-induced inhibition of gastrointestinal transit more potently than the inhibition induced by morphine or oxycodone. Naloxone methiodide suppressed the oxycodone-induced inhibition of gastrointestinal transit more potently than the inhibition induced by morphine, indicating that μ-opioid receptor agonists induce the inhibition of gastrointestinal transit through different mechanisms. Furthermore, we found that the route of administration (intracerebroventricular, intrathecally, and/or intraperitoneally) of naloxone methiodide differentially influenced the suppressive effect on the inhibition of colorectal transit induced by morphine, oxycodone, and fentanyl. These results suggest that morphine, oxycodone, and fentanyl induce constipation through different mechanisms (naloxonazine-sensitive versus naloxonazine-insensitive sites and central versus peripheral opioid receptors), and these findings may help us to understand the characteristics of the constipation induced by each μ-opioid receptor agonist and improve the quality of life by reducing constipation in patients being treated for pain.


Zn(II) Complex of Plant Phenolic Chlorogenic Acid: Antioxidant, Antimicrobial and Structural Studies.
The structure of the Zn(II) complex of 5-caffeoylquinic acid (chlorogenic acid, 5-CQA) and the type of interaction between the Zn(II) cation and the ligand were studied by means of various experimental and theoretical methods, i.e., electronic absorption spectroscopy UV/Vis, infrared spectroscopy FT-IR, elemental, thermogravimetric and density functional theory (DFT) calculations at B3LYP/6-31G(d) level. DPPH (2,2-diphenyl-1-picrylhydrazyl), ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), FRAP (ferric reducing antioxidant power), CUPRAC (cupric reducing antioxidant power) and trolox oxidation assays were applied in study of the anti-/pro-oxidant properties of Zn(II) 5-CQA and 5-CQA. The antimicrobial activity of these compounds against Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, Staphylococcus aureus, Salmonella enteritidis and Candida albicans was tested. An effect of Zn(II) chelation by chlorogenic acid on the anti-/pro-oxidant and antimicrobial activities of the ligand was discussed. Moreover, the mechanism of the antioxidant properties of Zn(II) 5-CQA and 5-CQA were studied on the basis of the theoretical energy descriptors and thermochemical parameters. Zn(II) chlorogenate showed better antioxidant activity than chlorogenic acid and commonly applied natural (L-ascorbic acid) and synthetic antioxidants (butylated hydroxyanisol (BHA) and butylated hydroxytoluene (BHT)). The pro-oxidant activity of Zn(II) 5-CQA was higher than the ligand and increased with the rise of the compound concentration The type of Zn(II) coordination by the chlorogenate ligand strongly affected the antioxidant activity of the complex.


Study on possible control algorithms for lower limb rehabilitation system.
In this paper, one of the possibilities of introduction of variable stiffness in a rehabilitation manipulator for lower limbs, is described. Mechanical design and working principle of a proposed system are presented. The following control algorithms for manipulators with flexible joints are suggested: the position control and the active impedance control. The tracking performance is experimentally studied for sinusoidal and polynomial reference trajectories. Obtained results confirm that both control strategies are suitable for controlling a rehabilitation manipulator with changeable stiffness.


Virtual reality simulators for gastrointestinal endoscopy training.
The use of simulators as educational tools for medical procedures is spreading rapidly and many efforts have been made for their implementation in gastrointestinal endoscopy training. Endoscopy simulation training has been suggested for ascertaining patient safety while positively influencing the trainees' learning curve. Virtual simulators are the most promising tool among all available types of simulators. These integrated modalities offer a human-like endoscopy experience by combining virtual images of the gastrointestinal tract and haptic realism with using a customized endoscope. From their first steps in the 1980s until today, research involving virtual endoscopic simulators can be divided in two categories: investigation of the impact of virtual simulator training in acquiring endoscopy skills and measuring competence. Emphasis should also be given to the financial impact of their implementation in endoscopy, including the cost of these state-of-the-art simulators and the potential economic benefits from their usage. Advances in technology will contribute to the upgrade of existing models and the development of new ones; while further research should be carried out to discover new fields of application.


Is evolution necessary for range expansion? Manipulating reproductive timing of a weedy annual transplanted beyond its range.
Ecologists often consider how environmental factors limit a species' geographic range. However, recent models suggest that geographic distribution also may be determined by a species' ability to adapt to novel environmental conditions. In this study, we empirically tested whether further evolution would be necessary for northern expansion of the weedy annual cocklebur (Xanthium strumarium) in its native North American range. We transplanted seedlings beyond the northern border and photoperiodically manipulated reproductive timing, a trait important for adaptation to shorter growing seasons at higher latitudes within the range, to determine whether further evolution of this trait would result in a phenotype viable beyond the range. Earlier reproductive induction enabled plants to produce mature seeds beyond the range and to achieve a reproductive output similar to those grown within the range. Therefore, evolution of earlier reproduction in marginal populations would be necessary for northward range expansion. This study is the first to empirically show that evolution in an ecologically important trait would enable a species to survive and reproduce beyond its current range. These results suggest that relatively few traits may limit a species' range and that identifying evolutionary constraints on such traits could be important for predicting geographic distribution.


Two cases of late onset Ota's naevus.
Ota's naevus is among the dermal melanocytoses that show a distinct pattern involving skin innervated by the trigeminal nerve. Most cases present at birth or manifest clinically in early childhood. Cases of acquired lesions in adult onset have been reported rarely. We present two cases of late onset Ota's naevus which were confirmed by skin biopsies. Both patients underwent Q-switched alexandrite laser treatment with a dose of 8.0 J/cm2 given four or five times at 6 weekly intervals and showed some improvement.


Multisite reproducibility and test-retest reliability of the T1w/T2w-ratio: A comparison of processing methods.
The ratio of T1-weighted (T1w) and T2-weighted (T2w) magnetic resonance imaging (MRI) images is often used as a proxy measure of cortical myelin. However, the T1w/T2w-ratio is based on signal intensities that are inherently non-quantitative and known to be affected by extrinsic factors. To account for this a variety of processing methods have been proposed, but a systematic evaluation of their efficacy is lacking. Given the dependence of the T1w/T2w-ratio on scanner hardware and T1w and T2w protocols, it is important to ensure that processing pipelines perform well also across different sites. We assessed a variety of processing methods for computing cortical T1w/T2w-ratio maps, including correction methods for nonlinear field inhomogeneities, local outliers, and partial volume effects as well as intensity normalisation. These were implemented in 33 processing pipelines which were applied to four test-retest datasets, with a total of 170 pairs of T1w and T2w images acquired on four different MRI scanners. We assessed processing pipelines across datasets in terms of their reproducibility of expected regional distributions of cortical myelin, lateral intensity biases, and test-retest reliability regionally and across the cortex. Regional distributions were compared both qualitatively with histology and quantitatively with two reference datasets, YA-BC and YA-B1+, from the Human Connectome Project. Reproducibility of raw T1w/T2w-ratio distributions was overall high with the exception of one dataset. For this dataset, Spearman rank correlations increased from 0.27 to 0.70 after N3 bias correction relative to the YA-BC reference and from -0.04 to 0.66 after N4ITK bias correction relative to the YA-B1+ reference. Partial volume and outlier corrections had only marginal effects on the reproducibility of T1w/T2w-ratio maps and test-retest reliability. Before intensity normalisation, we found large coefficients of variation (CVs) and low intraclass correlation coefficients (ICCs), with total whole-cortex CV of 10.13% and whole-cortex ICC of 0.58 for the raw T1w/T2w-ratio. Intensity normalisation with WhiteStripe, RAVEL, and Z-Score improved total whole-cortex CVs to 5.91%, 5.68%, and 5.19% respectively, whereas Z-Score and Least Squares improved whole-cortex ICCs to 0.96 and 0.97 respectively. In the presence of large intensity nonuniformities, bias field correction is necessary to achieve acceptable correspondence with known distributions of cortical myelin, but it can be detrimental in datasets with less intensity inhomogeneity. Intensity normalisation can improve test-retest reliability and inter-subject comparability. However, both bias field correction and intensity normalisation methods vary greatly in their efficacy and may affect the interpretation of results. The choice of T1w/T2w-ratio processing method must therefore be informed by both scanner and acquisition protocol as well as the given study objective. Our results highlight limitations of the T1w/T2w-ratio, but also suggest concrete ways to enhance its usefulness in future studies.


The alpha 1 Na(+)-K+ pump of the Dahl salt-sensitive rat exhibits altered Na+ modulation of K+ transport in red blood cells.
The properties of the alpha 1 Na(+)-K+ pump were compared in Dahl salt-sensitive (DS) and salt-resistant (DR) strains by measuring ouabain-sensitive fluxes (mmol/liter cell x hr = FU, Mean +/- SE) in red blood cells (RBCs) and varying internal (i) and external (o) Na+ and K+ concentrations. Kinetic parameters of several modes of operation, i.e., Na+/K+, K+/K+, Na+/Na+ exchanges, were characterized and analyzed for curve-fitting using the Enzfitter computer program. In unidirectional flux studies (n = 12 rats of each strain) into fresh cells incubated in 140 mM Na(+) + 5 mM K+, ouabain-sensitive K+ influx was substantially lower in the DS than in DR RBCs, while ouabain-sensitive Na+ efflux and Nai were similar in both strains. Thus, the coupling ratio between unidirectional Na+:K+ fluxes was significantly higher in DS than in DR cells at similar RBC Na+ content. In the presence of 140 mM Nao, activation of ouabain-sensitive K+ influx by Ko had a lower Km and Vmax in DS as estimated by the Garay equation (N = 2.70 +/- 0.33, Km 0.74 +/- 0.09 mM; Vmax 2.87 +/- 0.09 FU) than in DR rats (N = 1.23 +/- 0.36, Km 2.31 +/- 0.16 mM; Vmax 5.70 +/- 0.52 FU). However, the two kinetic parameters were similar following Nao removal. The activation of ouabain-sensitive K+ influx by Nai had significantly lower Vmax in DS (9.3 +/- 0.4 FU) than in DR (14.5 +/- 0.6 FU) RBCs but similar Km. These data suggest that the low K+ influx in DS cells is caused by a defect in modulation by Nao and Nai. Na+ efflux showed no differences in Nai activation or trans effects by Nao and Ko, thus accounting for the different Na+:K+ coupling ratio in the Dahl strains. Further evidence for the differences in the coupling of ouabain-sensitive fluxes was found in studies of net Na+ and K+ fluxes, where the net ouabain-sensitive Na+ losses showed similar magnitudes in the two Dahl strains while the net ouabain-sensitive K+ gains were significantly greater in the DR than the DS RBCs. Ouabain-sensitive Na+ influx and K+ efflux were also measured in these rat RBCs. The inhibition of ouabain-sensitive Na+ influx by Ko was fully competitive for the DS but not for the DR pumps. Thus, for DR pumps, Ko could activate higher K+ influx in DR pumps without a complete inhibition of ouabain-sensitive Na+ influx.(ABSTRACT TRUNCATED AT 400 WORDS)


Pharmacology of enkephalinase inhibitors: animal and human studies.
We have shown that a number of compounds which inhibit the degradation of met-enkephalin can produce naloxone-reversible analgesia in mice. These compounds also potentiate the analgesia produced by acupuncture, foot shock, and transcutaneous nerve stimulation in animals and humans. The potency of their effectiveness as analgesics or potentiators parallels their potency as inhibitors of mouse brain enkephalinase. D-Phenylalanine (DPA), one of these enkephalinase inhibitors, has been used successfully for the management of chronic intractable pain in humans and to potentiate the treatment of many painful conditions by acupuncture. Other aspects of pharmacology of DPA will be discussed, including its effects on the cardio-vascular system, behavior, and lack of development of tolerance and dependence when used chronically in animals and humans.


Interference by a phenylacetate pathway in isotopic assays for phenylalanine ammonia-lyase in leaf extracts.
Particulate and soluble fractions from leaves of Sorghum, Spinacia (spinach), and Coleus, capable of metabolizing l-phenylalanine to cinnamate or to caffeate, are also able to convert l-and d-phenylalanine to phenylacetate. Since cinnamate and phenylacetate are not effectively separated in commonly used chromatographic solvents, some of the isotropic assays used for phenylalanine ammonia-lyase are rendered ambiguous by the interference of this second pathway. Therefore, a "double decker," two-dimensional paper chromatographic method was designed to separate cinnamate and phenylacetate. This was combined with the use of phenylalanine labeled randomly or just in either the carbon 1 or 2 position of the side chain.


Time for a national approach to donor conception in Australia.
There is a growing recognition of the interests and rights of individuals conceived using donated gametes in assisted reproductive technology to information about their biological parentage. In Australia these rights vary between jurisdictions according to differing statutory provisions. In February 2011 the Senate's Legal and Constitutional Affairs References Committee published its report on Donor Conception Practices in Australia. The report recommended the development of a nationally consistent approach to donor conception and recommended the enactment of legislation in those Australian jurisdictions without legislation regulating donor conception. This editorial reviews the Senate Committee report and its recommendations and supports calls for a nationally harmonised approach to donor conception in Australia.


Intestinal microsporidiosis in human immunodeficiency virus-infected patients with chronic unexplained diarrhea: prevalence and clinical and biologic features.
Eighteen patients infected with human immunodeficiency virus and with chronic unexplained diarrhea were prospectively studied to investigate the prevalence and clinical and biologic features of intestinal microsporidiosis. All patients underwent extensive evaluation for bacterial, viral, and parasitic pathogens. Enterocytozoon bieneusi was found in 9 patients (50%; 95% confidence interval, 27-73) in stools and duodenal and jejunal biopsies. In 8 patients, it was the sole pathogen found. Other pathogens were also isolated from the intestinal tracts of 4 patients, but diarrhea remained unexplained in 6. Patients with intestinal microsporidiosis had significantly lower mean Karnofsky scores (69.4 vs. 85.5, P = .009), CD4 cell counts (18.6 vs. 209.8/microL, P = .02), and D-xylose absorption tests (0.13 vs. 0.36 g/L, P < .001) than did patients without intestinal microsporidiosis. Intestinal microsporidiosis appears to be a frequent cause of unexplained chronic diarrhea in patients with AIDS and is associated with diminished D-xylose absorption.


Case Report of Diffuse Large B Cell Lymphoma of Uterine Cervix Treated at a Semiurban Cancer Centre in North India.
Lymphoma of the uterine cervix is very rare. We report a case of diffuse large B cell lymphoma (DLBCL) involving the uterine cervix treated at a newly commissioned semiurban cancer centre in north India in 2015. Data for this study was obtained from the hospital electronic medical records and the patient's case file. We also reviewed published case reports of uterine and cervical lymphoma involving forty-one patients. We treated a case of stage IV DLBCL cervix with six cycles of R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisolone) and intrathecal methotrexate followed by consolidation with radiotherapy. The patient showed complete response to chemotherapy. We conclude that, in advanced stage lymphoma involving uterus and cervix, combination of chemotherapy and radiotherapy is effective in short term.


A review of the orthopedic interventions and functional outcomes among a cohort of 114 children with arthrogryposis multiplex congenita.
Arthrogryposis multiplex congenita (AMC) refers to a large heterogeneous group of conditions involving joint contractures in two or more different areas of the body. Contractures can lead to decreased range of motion and strength, and affect ambulation and autonomy. The aim of this study was to describe the orthopedic interventions and functional outcomes of a large cohort of children with AMC followed in a pediatric orthopedic center. A retrospective chart review of all children diagnosed with AMC followed at Shriners Hospital for Children - Canada (SHC) between January 1979 and July 2016 was conducted. One hundred twenty patients were identified, of whom six were excluded due to misdiagnosis or insufficient chart information. One hundred fourteen were retained. Patient demographics, AMC classification, comorbidities, operative and non-operative treatments received as well as community ambulation status, level of autonomy in self-care and transfers at latest follow-up were recorded. There were 54 males and 60 females with a mean age at last clinic visit of 10 years 3 months. Amyoplasia and distal arthrogryposis (DA) were equally represented in our sample, 47 (41.2%) and 49 (43.0%) participants respectively, with the category Other comprising the remaining 18 (15.8%) participants. Children with DA had less involvement of the proximal joints than those in the two other groups. Contractures and deformities of the foot and ankle were the most prevalent, affecting 91.5% with Amyoplasia, 85.7% with DA and 83.3% in the Other category. Contractures of the shoulder and elbow were more common among individuals with Amyoplasia and those categorized Other than those with DA. In terms of walking ability, 98% of participants with DA were independent ambulators. Walking ability varied among the Other participants. Similarly, most children with DA were independent in self-care and transfers at the most recent follow-up. The relatively large sample size of this study allowed for a better insight into the challenges associated with AMC management. These findings demonstrated the need for genetic testing to provide accurate diagnosis and classification, along with the use of standardized outcome tools to measure effectiveness of interventions. As AMC is rare, multi-site prospective studies are needed to improve research opportunities, develop functional measures specific to AMC and disseminate findings on a wider scale.


High rates of recurrence in HIV-infected and HIV-uninfected patients with tuberculosis.
The rate of recurrent tuberculosis disease due to reinfection, compared with the incidence of new tuberculosis, in those with and without HIV infection is not known. In a retrospective cohort study of South African gold miners, men with known dates of seroconversion to HIV (from 1991 to 1997) and HIV-negative men were followed up to 2004. Rates of tuberculosis recurrence >2 years after the first episode were used as a proxy for reinfection disease rates. Among 342 HIV-positive and 321 HIV-negative men who had had 1 previous episode of tuberculosis, rates of recurrence were 19.7 cases per 100 person-years at risk (PYAR; 95% confidence interval [CI], 16.4-23.7) and 7.7 cases per 100 PYAR (95% CI, 6.1-9.8), respectively. The recurrence rate did not vary by duration of HIV infection. Recurrent pulmonary tuberculosis rates >2 years after the first episode were 24.4 cases per 100 PYAR (95% CI, 17.2-34.8) in HIV-positive men and 4.3 cases per 100 PYAR (95% CI, 2.2-8.3) in HIV-negative men, compared with incidence rates of new pulmonary tuberculosis of 3.7 cases per 100 PYAR (95% CI, 3.3-4.1) in HIV-positive men and 0.75 cases per 100 PYAR (95% CI, 0.67-0.84) in HIV-negative men in the same cohort. Tuberculosis recurrence rates, likely due to reinfection, were much higher than incidence rates. The findings suggest heterogeneity in susceptibility, implying that a vaccine could still provide useful protection in the population and strengthening the case for secondary preventive therapy.


Validity of the utrecht scale for evaluation of rehabilitation-participation.
There is still a need for a generic participation instrument that measures both objective and subjective participation in adults living in the community and that is feasible for use in rehabilitation practice. The Utrecht Scale for Evaluation of Rehabilitation-Participation (USER-Participation) was developed to satisfy this need, comprising 31 items in three scales: Frequency, Restrictions and Satisfaction. The aim of this study was to examine the validity of this measure. Cross-sectional study involving former rehabilitation outpatients from five rehabilitation facilities in the Netherlands (n = 395). The Frenchay Activities Index (FAI), the participation subtotal score of the ICF Measure of Participation and Activities Screener (IMPACT-SP) and the Participation Scale were included as reference measures. Internal consistency of the USER-Participation scales was satisfactory (α 0.70-0.91). Spearman correlations between these scales were between 0.36 and 0.52. Concurrent validity was shown by strong correlations between the Frequency scale and the FAI (0.59), the Restrictions scale and the IMPACT-SP (0.75) and the Satisfaction scale and the Participation Scale (-0.73). Discriminant validity was shown by significant differences in USER-Participation scores between participants with different levels of independence and between participants with different health conditions. The USER-Participation appears to be a valid measure to rate objective and subjective participation in persons with physical disabilities.


Impacts of Simulated Weightlessness by Dry Immersion on Optic Nerve Sheath Diameter and Cerebral Autoregulation.
Dry immersion (DI) is used to simulate weightlessness. We investigated in healthy volunteers if DI induces changes in ONSD, as a surrogate marker of intracranial pressure (ICP) and how these changes could affect cerebral autoregulation (CA). Changes in ICP were indirectly measured by changes in optic nerve sheath diameter (ONSD). 12 healthy male volunteers underwent 3 days of DI. ONSD was indirectly assessed by ocular ultrasonography. Cerebral blood flow velocity (CBFV) of the middle cerebral artery was gauged using transcranial Doppler ultrasonography. CA was evaluated by two methods: (1) transfer function analysis was calculated to determine the relationship between mean CBFV and mean arterial blood pressure (ABP) and (2) correlation index Mxa between mean CBFV and mean ABP.ONSD increased significantly during the first day, the third day and the first day of recovery of DI (P < 0.001).DI induced a reduction in Mxa index (P < 0.001) and an elevation in phase shift in low frequency bandwidth (P < 0.05). After DI, Mxa and coherence were strongly correlated with ONSD (P < 0.05) but not before DI. These results indicate that 3 days of DI induces significant changes in ONSD most likely reflecting an increase in ICP. CA was improved but also negatively correlated with ONSD suggesting that a persistent elevation ICP favors poor CA recovery after simulated microgravity.


[Antibiotic sensitivity and other properties of bacteria of the genus Aeromonas].
Antibiotic sensitivity and other biological properties of Aeromonas and NAD-vibrios isolated from water of open waterbodies were studied comparatively. It was found that the organisms of these genera had a definite similarity with respect to antibiotic sensitivity, morphological, cultural and some antigenic properties. The differences in the antibiotic sensitivity of these organisms were not sufficient for definition of the taxonomic features.


Association between sleep duration and chest pain in US adults: A cross-sectional study.
Herein, we purposed to explore the association of sleep duration with chest pain among adults in US. This research work enrolled 13,274 subjects in the National Health and Nutrition Examination Survey (NHANES) from 2011 to 2018. The association of sleep duration with chest pain among adults in US was evaluated by Multivariable logistic regression. To elucidate the association, we made adjustments for gender, BMI, diabetes, smoking status, drinking status, race, marital status, annual family income, hyperlipoidemia, Hypertension. Chest pain incidence decreased by 5% [OR = 0.95 (0.93, 0.98), p = 0.0004] for an increase in sleep duration by 1 h. A generalized additive model (GAM) was used to reseal a U-shaped relationship of sleep duration with incident chest pain. When duration of sleep was <6.5 h, chest pain incidence negatively correlated to sleep duration [OR = 0.77 (0.72, 0.82) P < 0.0001]. However, when sleep duration was ≥6.5 h, chest pain incidence rose with escalating sleep duration [OR = 1.07 (1.03, 1.11) p = 0.0014]. Duration of sleep was established to be independently linked with an increase in the occurrence of chest pain. Excessive sleep, as much as insufficient sleep, increases the risk of chest pain. Both excessive sleep and insufficient sleep are associated with an increased risk of chest pain.


Pluronic/chitosan hydrogels containing epidermal growth factor with wound-adhesive and photo-crosslinkable properties.
Wound-adhesive and thermo-responsive hydrogels were prepared in an aim to develop a wound-care device for diabetic ulcers. A mixture of glycidyl methacrylated chitooligosaccharide (COS), di-acrylated Pluronic, and a recombinant human epidermal growth factor (rhEGF) formed a physical hydrogel. Photo-irradiation was subsequently applied to chemically crosslink the hydrogel. Release profiles of encapsulated rhEGF from the hydrogel showed that release rates were dependent on degradation rates of hydrogels. Human primary keratinocytes were cultivated with the released fraction from the hydrogel to measure effects of released rhEGF on in vitro differentiation of keratinocytes. Muco-adhesive property of the hydrogel was investigated in animal skins and significant amounts of quantum dot-labeled rhEGF were retained at wound sites along with chitosan. The hydrogel was administered to dorsal burn wounds in wound healing impaired animals and photo-irradiated. COS and rhEGF in the hydrogels significantly enhanced epidermal differentiation during the wound healing process. Thus, the rhEGF-encapsulated hydrogel was expected to be a potential wound care product by increasing local concentration of rhEGF at wound sites with maintaining keratinocytic differentiation.


Comparison of accessory olfactory bulb volumes in the common tree shrew (Tupaia glis).
The volume size of the accessory olfactory bulb (AOB) of 40 common tree shrews (Tupaia glis) was compared with regard to differences between left and right sides, males and females, and animals born in the wild and those born and raised in captivity. There were no statistically significant differences between the two sides and the two sexes, but a significant reduction of the AOB from wild to captive animals was apparent. This reduction was more pronounced in females than in males and somewhat more pronounced in the inner granular layer (layer 6) than in the other measured components (layers 1 + 2, layers 3-5). No well-founded explanation for this reduction could be given.


Conformational changes of the nucleotide site of the plasma membrane Ca2+-ATPase probed by fluorescence quenching.
Fluorescence quenching by the water-soluble ions I(-) and Cs(+) was used to probe solvent accessibility and polarity of the nucleotide/fluorescein isothiocyanate binding pocket of the purified soluble Ca(2+)-ATPase from plasma membranes. The E(1).Ca.CaM conformer was the least accessible state studied, presenting the lowest suppression constant (K(q)) for both I(-) (K(q) = 6.7 M(-)(1)) and Cs(+) (K(q) = 0.7 M(-)(1)). Accessibility to I(-) was similar for the E(2).VO(4) and E(1).Ca states (K(q) = 7.13 and 7.5 M(-)(1), respectively), whereas E(2) was slightly more accessible (K(q) = 9.1 M(-)(1)). The phosphorylated state E(2)-P presented the highest accessibility, with a K(q) of 16.5 M(-)(1), very near the K(q) of 20.3 M(-)(1) for free FITC. I(-) was unequivocally a better fluorescence quencher, being usually nearly 3-fold as efficient as Cs(+), as indicated by the K(q)(I(-))/K(q)(Cs(+)) ratio (R(q)). The advent of a positive charge cluster on the nucleotide/fluorescein binding pocket in different states was suggested by the increase in R(q), which reached a value as high as 9.5 for the E(1).Ca.CaM conformer. These results indicate (i) a very high water accessibility of the nucleotide/fluorescein pocket for E(2)-P that (ii) is more restricted on the free E(2) state and (iii) becomes rather lower for the E(1).Ca states. Additionally, a positive charge effect of amino acids on the nucleotide site, possibly related to ATP binding and phosphoryl transfer, appears in these E(1).Ca states, being absent in the phosphorylated and nonphosphorylated E(2) states.


Early adaptive humoral immune responses and virus clearance in humans recently infected with pandemic 2009 H1N1 influenza virus.
Few studies on the humoral immune responses in human during natural influenza infection have been reported. Here, we used serum samples from pandemic 2009 H1N1 influenza infected patients to characterize the humoral immune responses to influenza during natural infection in humans. We observed for the first time that the pandemic 2009 H1N1 influenza induced influenza A-specific IgM within days after symptoms onset, whereas the unit of IgG did not changed. The magnitude of influenza A-specific IgM antibodies might have a value in predicting the rate of virus clearance to some degree. However, the newly developed IgM was not associated with hemagglutination inhibition (HI) activities in the same samples but correlated with HI activities of subsequently collected sera which were mediated by IgG antibodies, indicating that IgM was critical for influenza infection and influences subsequent IgG antibody responses. These findings provide new important insights on the human immunity to natural influenza infection.


Tuning the Electrocatalytic Oxygen Reduction Reaction Activity and Stability of Shape-Controlled Pt-Ni Nanoparticles by Thermal Annealing - Elucidating the Surface Atomic Structural and Compositional Changes.
Shape-controlled octahedral Pt-Ni alloy nanoparticles exhibit remarkably high activities for the electroreduction of molecular oxygen (oxygen reduction reaction, ORR), which makes them fuel-cell cathode catalysts with exceptional potential. To unfold their full and optimized catalytic activity and stability, however, the nano-octahedra require post-synthesis thermal treatments, which alter the surface atomic structure and composition of the crystal facets. Here, we address and strive to elucidate the underlying surface chemical processes using a combination of ex situ analytical techniques with in situ transmission electron microscopy (TEM), in situ X-ray diffraction (XRD), and in situ electrochemical Fourier transformed infrared (FTIR) experiments. We present a robust fundamental correlation between annealing temperature and catalytic activity, where a ∼25 times higher ORR activity than for commercial Pt/C (2.7 A mgPt-1 at 0.9 VRHE) was reproducibly observed upon annealing at 300 °C. The electrochemical stability, however, peaked out at the most severe heat treatments at 500 °C. Aberration-corrected scanning transmission electron microscopy and energy-dispersive X-ray spectroscopy (EDX) in combination with in situ electrochemical CO stripping/FTIR data revealed subtle, but important, differences in the formation and chemical nature of Pt-rich and Ni-rich surface domains in the octahedral (111) facets. Estimating trends in surface chemisorption energies from in situ electrochemical CO/FTIR investigations suggested that balanced annealing generates an optimal degree of Pt surface enrichment, while the others exhibited mostly Ni-rich facets. The insights from our study are quite generally valid and aid in developing suitable post-synthesis thermal treatments for other alloy nanocatalysts as well.


Lumbar puncture-related cerebrospinal fluid leakage on magnetic resonance myelography: is it a clinically significant finding?
Post-dural puncture headache (PDPH) due to excessive cerebrospinal fluid (CSF) leakage is a well-known complication of lumbar puncture. Although various factors, especially the type of spinal needle, have been demonstrated to be associated with PDPH, the clinical implications of CSF leakage detected on magnetic resonance myelography (MRM) images remain unclear. The objective of this case-control study was to evaluate the association between radiologically visualized CSF leakage and PDPH. Clinical data including patients' age and gender, types of spinal needle, duration of bed rest, interval between lumbar puncture procedures and MRM studies, and incidence of PDPH were compared between patients who were radiologically-positive and -negative for CSF leakage. Of the 22 patients with definite CSF leakage on MRM images, most were asymptomatic (86%, 19/22). The remaining three patients, who were suffering from PDPH, only complained of headaches and were treated conservatively. In a review of patients' clinical data, there were no significant differences in any parameter including the incidence of PDPH between the 22 patients who were radiologically-positive for CSF leakage and the 31 radiologically-negative patients. The significance of radiologically visualized CSF leakage should not be overestimated, as most such incidents are not associated with PDPH and do not require any treatment.


Adolescent smoking cessation methods: a review article.
The rate at which adolescents in West Virginia are smoking continues to be an ongoing concern. Approximately 9% of middle school and 22% of high school students are smoking cigarettes. Although these figures have been on a decline, this still represents a staggering figure. Review the literature and research on tobacco smoking cessation programs available to adolescents. The primary goal was to identify and review studies aimed at adolescent tobacco cessation that have been performed over the last decade. Journal articles from PubMed, Cochrane Database, Centers for Disease control (CDC) website, and West Virginia Department of Health & Human Resources (WVDHHR) website were used to capture recently published data.


Judgment, 26 April 1988.
The Court granted the petition sought by the Juvenile Defender of the Argentina Ministry of Minors to prevent a mentally retarded minor who had been the victim of rape and who was four months into her pregnancy from having an abortion. The Defender sought to prevent the abortion because he believed 1) that the fetus had a right to life and 2) that the performance of the abortion would pose a significant health risk to the mother. Although the mother of the minor had consented under provisions of the Penal Code allowing a pregnancy occurring under such circumstances to be terminated if the legal representative of the mentally retarded woman consents, the Court ruled that there was no consent. It concluded that the mother was not the legal representative of the minor because the minor did not live with her parents and had not been legally declared mentally incompetent. The Court also stated that, in cases where a pregnant woman with parental authority petitions to have an abortion, the law mandates that the fetus be represented legally and that the court must consult the Ministry of Minors before it authorizes an abortion. It based its decision on the legal right of a fetus to life.


Interaction of the calcium channel activating 3H-BAY K 8644 and inhibiting 3H-verapamil with specific receptor sites on cultured beating myocardial cells.
Binding properties of the calcium channel activating dihydropyridine (DHP), H-BAY K 8644, and the inhibiting 3H-verapamil were demonstrated in monolayer cultures of beating cardiac cells. 3H-BAY K 8644 specific binding was dependent on the presence of extracellular calcium, the affinity was modulated by Ca2+, but Hill coefficients remained unaffected. BAY K 8644 stimulated myocardial contractility in resting and beating myocytes. In contrast to beta-adrenoceptor agonists, however, cellular levels of cyclic AMP and cyclic GMP in cultured myocytes remained unchanged by the compound. Dihydropyridine derivatives of both the calcium channel activating BAY K 8644 as well as the Ca2+ entry blocking DHPs of the nifedipine or nimodipine type yielded very low affinity to other receptors measured in brain and heart membranes. 3H-BAY K 8644 binding sites proved to be highly specific for various potently displacing DHP derivatives and discriminated between optical isomers (stereoselectivity) with inhibition constants (Ki) in the nanomolar range. The heterogeneous shapes of the competition curves also imply interactions of these compounds with different (sub-)sites of the DHP receptor that represents one locus of interaction in regulating transmembranal Ca2+ currents. The other specific site of action for the potent diphenylalkylamines, clearly different to the DHP receptor was characterized with 3H-verapamil. The equilibrium dissociation constant, Kd in cultured myocytes ranged between 16-25 nM, and binding capacity, Bmax amounted to about 1.85 pmol/mg of protein. The different mode of competitions indicates the involvement of more than one 3H-verapamil binding site. The interrelation of the structurally heterogeneous channel modulators with the differently radiolabelled receptor (sub-)sites located in or near by the calcium channel may represent new approaches in investigating the nature of action of these potent compounds.


Surfing Behind a Boat: Quality and Reliability of Online Resources on Scaphoid Fractures.
Patients seeking information and advice on treatment of scaphoid fractures unknowingly confront longstanding medical controversies surrounding the management of this condition. However, there are no studies specifically looking into the quality and reliability of online information on scaphoid fractures. We identified 44 unique websites for evaluation using the term "scaphoid fractures". The websites were categorized by type and assessed using the DISCERN score, the Journal of the American Medical Association (JAMA) benchmark criteria and the Health on the net (HON) code. The majority of websites were commercial (n = 13) followed by academic (n = 12). Only seven of the websites were HON certified. The mean DISCERN score was 43.8. Only 4 websites scored 63 or above representing excellent quality with minimal shortcomings but 13 websites scored 38 or below representing poor or very poor quality. The mean JAMA benchmark criteria score was 2.2. The Governmental and Non-Profit Organizations category websites had the highest mean JAMA benchmark score. The websites that displayed the HON-code seal had higher mean DISCERN scores and higher mean JAMA benchmark scores compared to websites that did not display the seal. Good quality health information is certainly available on the Internet. However, it is not possible to predict with certainty which sites are of higher quality. We suggest clinicians should have a responsibility to educate their patients regarding the unregulated nature of medical information on the internet and proactively provide patients with educational resources and thus help them make smart and informed decisions.


Guidelines for point-of-care testing. Improving patient outcomes.
Whole-blood biosensors and point-of-care testing created a unique paradigm in medical diagnostics in the 1980s, when liver and heart transplant centers, as well as operating rooms and other critical care areas implemented whole-blood analysis to provide rapid test results in 2 to 5 minutes. Rising expectations, patient-focused hospitals, and managed care intensify the need for immediate decisions at the point of care. The guidelines promote consensus priorities, multidisciplinary teamwork, fiscal coordination, and collaborative practice during this phase of rapid change. Four primary principles and accompanying guideline objectives are optimization (patient outcomes, medical linkages, integrated diagnostic synthesis, therapeutic turnaround time, test clusters, and critical limits), hybridization (strategic modalities, economic effectiveness, and testing --> monitoring), quality (quality improvement, clinical performance, risk reduction and academics and accreditation), and consistency (results communication and error minimization, and reference intervals and standardization). Whole-blood analysis and point-of-care testing help facilitate temporal optimization, optimize diagnostic-therapeutic processes, and improve patient outcomes in critical care.


Resistant to Targeted Therapy - Aim for Metabolic Liabilities.
The advent of targeted therapies generated much optimism when discovered. Targeted therapies, are however associated with rapid acquisition of resistance. In a recent study by Dong et al. (Theranostics 2018; 8(7):1808-1823. doi:10.7150/thno.23177) it was shown that lung tumors resistant to the EGFR-inhibitor (Erlotinib), reprogram their metabolism and acquire a pro-survival dependency on Phosphoglycerate Dehydrogenase (PHGDH) that can be targeted to eliminate resistant tumors.


SOCE-inhibitor reduced human sperm-induced formation of neutrophil extracellular traps.
Human spermatozoa activate neutrophil extracellular traps (NETs) in vitro. NETosis is an efficient mechanism through which polymorphonuclear neutrophils (PMN) capture sperm in vitro. The objective of this study was to establish the role of store-operated Ca+2 entry (SOCE) in human sperm-triggered NETs and its impact on sperm integrity and oocyte binding capacity. PMN isolated from donors were exposed to spermatozoa isolated from normozoospermic donors using the swim-up technique and were divided into the following groups: (1) sperm, (2) PMN, (3) PMN + sperm, (4) PMN (pretreated with 2-APB, SOCE inhibitor) + sperm, (5) (PMN + DNase) + sperm, and (6) (PMN + PMA) + sperm (positive control). NETs were quantified using PicoGreen® and visualised by scanning electron microscopy and immunofluorescence of extracellular DNA and neutrophil elastase. Plasma membrane, acrosome, and DNA integrity were analysed by flow cytometry, and oocyte binding was evaluated using the hemizona pellucida assay. Sperm-triggered NETosis negatively affected the sperm membrane and acrosome integrity and decreased the oocyte binding capacity. These effects were negated by an SOCE inhibitor, thus improving sperm function and achieving high oocyte binding capacity. The SOCE inhibitor significantly reduced NET formation compared with that in control PMN/sperm (P < 0.05). Collectively, these results advance the knowledge about the role of PMN in reproduction and will allow the development of strategies to block NET formation in situations of reduced fertilisation success.


Relationship between carotid intima thickness and silent cerebral infarction in patients with type 2 diabetic nephropathy.
To examine the relationship between carotid intima media thickness (CIMT) and silent cerebral infarction (SCI) in order to determine whether CIMT is a predictor of SCI in patients of type 2 diabetic nephropathy. A total of 80 patients of type 2 diabetic nephropathy were selected on the basis of fasting and 2-hour post-prandial blood sugar, 24 hrs albumin estimation in urine, urea and creatinine in the serum. The selected candidates underwent MRI brain and carotid B mode ultrasonography to find out the event of SCI and to evaluate the CIMT respectively. The SCI was found in 30 (37.5%) patients with type 2 diabetic nephropathy. The mean age, BMI, blood pressure (BP), macroalbuminuria, S. lipids, low GFR, duration of diabetes and CIMT were significantly higher in the subject with SCI than in those without it. Multiple logistic analyses indicated that age, BP, and CIMT were found to be significant and independent risk factors of SCI in type 2 diabetic nephropathy subjects. CIMT is a surrogate and reliable predictor of higher risk of SCI among type 2 diabetic nephropathy patients.


Anti-inflammatory mediators as physiological and pharmacological regulators of parturition.
Increasing evidence highlights parturition as an inflammatory event characterized by leukocyte influx and proinflammatory mediator production in the intrauterine environment. While the mechanisms responsible for the initiation of this inflammatory cascade are not well understood, it is clear that these inflammatory events must be tightly regulated as the premature activation of these inflammatory signals is associated with adverse pregnancy outcomes, such as preterm labor, which is the leading cause of neonatal mortality and morbidity. In this article we highlight the importance of anti-inflammatory factors in regulating the inflammatory events surrounding parturition and discuss the use of anti-inflammatory mediators as potential novel therapeutic agents in the treatment of inflammation-induced preterm labor.


Our experience with dermofat graft in reconstruction of anophthalmic socket.
To present our experience with dermofat graft in reconstruction of anophthalmic socket. In period from September 2005 until June 2009, eight patients have undergone orbital surgery of transplantation of dermofat graft. In six patients the dermofat graft was used as the secondary orbital implant after extrusion of hydroxyapatite orbital implant with major defect of bulbar conjunctiva. The other indication for the dermofat graft was correction of deep superior sulcus of the upper lid in anophthalmic socket in two patients. The graft was harvested from the left side of the belly. The size of the graft purposely exceeded the size of the defect to account for the expected tissue resorption. We experienced no major complication. Approximately 20-40% of dermofat graft reduction was noticed in 3-months period postoperatively. It took 6-8 weeks for the graft to fully epithelize from the conjunctival edge. Silicone conformer was introduced for that period of time. Subsequently, in two out of six patients with dermofat graft as the secondary implant, fornix had to be reconstructed later on with oral mucosa graft. Dermofat graft is a valuable material in orbital reconstruction especially in anophthalmic socket.


Characterization of the interface between gamma and epsilon subunits of Escherichia coli F1-ATPase.
The interaction faces of the gamma and epsilon subunits in the Escherichia coli F1-ATPase have been explored by a combination of cross-linking and chemical modification experiments using several mutant epsilon subunits as follows: epsilonS10C, epsilonH38C, epsilonT43C, epsilonS65C, epsilonS108C, and epsilonM138C, along with a mutant of the gamma subunit, gammaT106C. The replacement of Ser-10 by a Cys or Met-138 by a Cys reduced the inhibition of ECF1 by the epsilon subunit, while the mutation S65C increased this inhibitory effect. Modification of the Cys at position 10 with N-ethylmaleimide or fluoroscein maleimide further reduced the binding affinity of, and the maximal inhibition by, the epsilon subunit. Similar chemical modification of the Cys at position 43 of the epsilon subunit (in the mutant epsilonT43C) and a Cys at position 106 of the gamma subunit (gammaT106C) also affected the inhibition of ECF1 by the epsilon subunit. The various epsilon subunit mutants were reacted with TFPAM3, and the site(s) of cross-linking within the ECF1 complex was determined. Previous studies have shown cross-linking from the Cys at positions 10 and 38 with the gamma subunit and from a Cys at position 108 to an alpha subunit (Aggeler, R., Chicas-Cruz, K., Cai, S. X., Keana, J. F. W., and Capaldi, R. A. (1992) Biochemistry 31, 2956-2961; Aggeler, R., Weinreich, F., and Capaldi, R. A. (1995) Biochim. Biophys. Acta 1230, 62-68). Here, cross-linking was found from a Cys at position 43 to the gamma subunit and from the Cys at position 138 to a beta subunit. The site of cross-linking from Cys-10 of epsilon to the gamma subunit was localized by peptide mapping to a region of the gamma subunit between residues 222 and 242. Cross-linking from a Cys at position 38 and at position 43 was with the C-terminal part of the gamma subunit, between residues 202 and 286. ECF1 treated with trypsin at pH 7.0 still binds purified epsilon subunit, while enzyme treated with the protease at pH 8.0 does not. This identifies sites around residue 70 and/or between 202 and 212 of the gamma subunit as involved in epsilon subunit binding.


Pharmacological evidence supporting a role for central corticotropin-releasing factor(2) receptors in behavioral, but not endocrine, response to environmental stress.
Corticotropin-releasing factor (CRF) is one of the principle components of the stress response. The physiological effects of CRF are mediated by two receptor subtypes, CRF(1) and CRF(2). Recent data obtained with the selective CRF(2) antagonist antisauvagine-30 (ASV-30) has begun to suggest that both CRF receptor subtypes may play a role in stress-related behaviors. Exactly how these two receptor subtypes interact to modulate the behavioral and endocrine responses to stress is not clear, however. We have attempted to understand the role of the CRF(2) receptor in the behavioral and endocrine responses to stress by comparing the effects of ASV-30 with the mixed CRF(1)/CRF(2) receptor antagonist astressin. Centrally administered ASV-30 reduced anxiety-like behavior in BALB/c mice in three models of anxiety: marble burying [minimal effective dose (MED) = 3 nmol], open field (MED = 3 nmol), and elevated plus maze (MED = 0.1 nmol). ASV-30 did not change locomotor activity or the adrenocorticotropic hormone (ACTH) response to restraint stress. The potent mixed CRF(1)/CRF(2) antagonist astressin not only reduced anxiety-like behavior in all three models with equivalent potency but also blunted the ACTH response to restraint stress. Finally, the new selective CRF(2) receptor agonist urocortin-II produced a dose-dependent increase in anxiety-like behavior in the plus maze test. Therefore, our data suggest that the CRF(2) receptor plays a role in the behavioral, but not the hypothalamic-pituitary-adrenal axis, response to stress.


Mutagenesis and DNA adduct formation in the mouse mammary gland exposed to 2-hydroxyamino-1-methyl-6-phenylimidazo-[4,5-b]pyridine in whole organ culture.
2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is a mutagen and rodent mammary gland carcinogen found in the human diet. 2-Hydroxyamino-1-methyl-6-phenylimidazo[4,5-b]pyridine (N-hydroxy-PhIP) is the proximate reactive metabolite of PhIP associated with PhIP-DNA adduct formation and mutagenesis. In the current study, whole mammary glands obtained from transgenic C57Bl/6 mice carrying the plasmid-lacZ mutational reporter gene were cultured in defined medium and exposed to various concentrations of N-hydroxy-PhIP for 24 h. At various times after N-hydroxy-PhIP exposure, PhIP-DNA adduct levels were determined by the (32)P-post-labeling assay and the lacZ(-) mutant frequency determined by the positive selection system. Glands were cultured in either medium containing insulin (I medium), necessary for maintenance of the gland, or I medium containing prolactin, aldosterone and hydrocortisone (IPAH medium) to induce lobuloalveolar development. At 3 and 7 days after exposure to 10 micro M N-hydroxy-PhIP, mutant frequency was upwards of 9-fold higher in glands incubated in IPAH medium than in I medium (15.2 +/- 1.9 and 1.6 +/- 0.7 x 10(-3), respectively, 3 day time point). PhIP-DNA adduct levels were 1.7-fold higher in glands cultivated in IPAH medium than in I medium immediately after exposure to 10 micro M N-hydroxy-PhIP. A statistically significant reduction in PhIP-DNA adduct levels occurred with time in glands cultivated in IPAH medium but not I medium (one-way analysis of variance, P < 0.05). By 7 days after exposure, PhIP-DNA adduct levels were similar in glands cultured in I and IPAH medium (3.2 +/- 0.2 and 2.8 +/- 0.29 adducts/10(7) nucleotides, respectively). DNA synthesis as measured by [(3)H]thymidine labeling was approximately 2-fold higher in glands cultured in IPAH medium than in I medium. The higher mutant frequency in glands cultivated in IPAH medium versus I medium appeared to be due to a combination of higher initial PhIP-DNA adduct levels and a greater fixation of mutations that occurred at higher proliferation rates. The findings indicate that mammotrophic hormones influence the mutagenicity of PhIP in the mammary gland in vitro and emphasize the importance of hormonal milieu on carcinogen-DNA adduct-induced mutations in this organ.


A mild and chemoselective method for the reduction of conjugated isoxazolines to beta-hydroxy ketones.
A new procedure for the selective reduction of conjugated Delta(2)-isoxazolines to the corresponding unsaturated beta-hydroxy ketones is described. The use of SmI(2) as the reducing agent and B(OH)(3) to hydrolyze the resulting imine results in a mild, convenient, and chemoselective protocol for this otherwise difficult transformation and complements existing methodology for the preparation of beta-hydroxy ketones via nitrile oxides.


Thermal stability of rhodopsin and progression of retinitis pigmentosa: comparison of S186W and D190N rhodopsin mutants.
Over 100 point mutations in the rhodopsin gene have been associated with retinitis pigmentosa (RP), a family of inherited visual disorders. Among these, we focused on characterizing the S186W mutation. We compared the thermal properties of the S186W mutant with another RP-causing mutant, D190N, and with WT rhodopsin. To assess thermal stability, we measured the rate of two thermal reactions contributing to the thermal decay of rhodopsin as follows: thermal isomerization of 11-cis-retinal and hydrolysis of the protonated Schiff base linkage between the 11-cis-retinal chromophore and opsin protein. We used UV-visible spectroscopy and HPLC to examine the kinetics of these reactions at 37 and 55 °C for WT and mutant rhodopsin purified from HEK293 cells. Compared with WT rhodopsin and the D190N mutant, the S186W mutation dramatically increases the rates of both thermal isomerization and dark state hydrolysis of the Schiff base by 1-2 orders of magnitude. The results suggest that the S186W mutant thermally destabilizes rhodopsin by disrupting a hydrogen bond network at the receptor's active site. The decrease in the thermal stability of dark state rhodopsin is likely to be associated with higher levels of dark noise that undermine the sensitivity of rhodopsin, potentially accounting for night blindness in the early stages of RP. Further studies of the thermal stability of additional pathogenic rhodopsin mutations in conjunction with clinical studies are expected to provide insight into the molecular mechanism of RP and test the correlation between rhodopsin's thermal stability and RP progression in patients.


Formation of Monofluorinated Radical Cofactor in Galactose Oxidase through Copper-Mediated C-F Bond Scission.
Galactose oxidase (GAO) contains a Cu(II)-ligand radical cofactor. The cofactor, which is autocatalytically generated through the oxidation of the copper, consists of a cysteine-tyrosine radical (Cys-Tyr•) as a copper ligand. The formation of the cross-linked thioether bond is accompanied by a C-H bond scission on Tyr272 with few details known thus far. Here, we report the genetic incorporation of 3,5-dichlorotyrosine (Cl2-Tyr) and 3,5-difluorotyrosine (F2-Tyr) to replace Tyr272 in the GAOV previously optimized for expression through directed evolution. The proteins with an unnatural tyrosine residue are catalytically competent. We determined the high-resolution crystal structures of the GAOV, Cl2-Tyr272, and F2-Tyr272 incorporated variants at 1.48, 1.23, and 1.80 Å resolution, respectively. The structural data showed only one halogen remained in the cofactor, indicating that an oxidative carbon-chlorine/fluorine bond scission has occurred during the autocatalytic process of cofactor biogenesis. Using hydroxyurea as a radical scavenger, the spin-coupled hidden Cu(II) was observed by EPR spectroscopy. Thus, the structurally defined catalytic center with genetic unnatural tyrosine substitution is in the radical containing form as in the wild-type, i.e., Cu(II)-(Cl-Tyr•-Cys) or Cu(II)-(F-Tyr•-Cys). These findings illustrate a previously unobserved C-F/C-Cl bond cleavage in biology mediated by a mononuclear copper center.


The relationship of skin test positivity, high serum total IgE levels, and peripheral blood eosinophilia to symptomatic and asymptomatic airway hyperresponsiveness.
The relationships of skin test positivity, high serum total IgE levels (> 100 kU/L), and peripheral blood eosinophilia (>/= 275 cells/microliter) to symptomatic (either chronic cough, chronic phlegm, bronchitis episodes, dyspnea, wheeze, or asthma) and asymptomatic bronchial hyperresponsiveness (BHR) were studied cross-sectionally in 620 adult subjects who participated in the Vlagtwedde-Vlaardingen Study of 1989 and 1990. Eosinophilia (OR = 2.06, 95% CI = 1.28 to 3.31) and skin test positivity (OR = 1.66, 95% CI = 1.02 to 2.71) were both significantly associated with BHR independent of age, sex, smoking, and urban area of residence. High serum total IgE levels were not associated with BHR (OR = 1.29, 95% CI = 0.81 to 2.03). Separate analyses for symptomatic and asymptomatic subjects showed that the higher risk of BHR with skin test positivity applied only to symptomatic subjects (OR = 5.78, 95% CI = 1.63 to 20.51), independent of eosinophilia and high serum total IgE levels. The higher risk of BHR with eosinophilia was not different between symptomatic and asymptomatic subjects, and independent of skin test positivity and high serum total IgE levels. The results of this study show that, in the general adult population, eosinophilia is associated with BHR both in symptomatic and asymptomatic persons, whereas skin test positivity is associated with BHR only in symptomatic subjects.


Creutzfeldt-Jakob disease and lyophilised dura mater grafts: report of two cases.
Two further cases of Creutzfeldt-Jakob disease (CJD) in association with cadaveric dura mater grafts are described. The clinical features of all such reported cases resemble more closely those of sporadic CJD, in contrast with kuru and the cases of CJD which have arisen after therapy with human pituitary-derived growth hormone. This observation may reflect the route of inoculation of the agent.