People With Cancer Experience Worse Psychosocial and Financial Consequences of COVID-19 Compared With Other Chronic Disease Populations: Findings From the International COVID-19 Awareness and Response Evaluation Survey Study.
The COVID-19 pandemic is likely to have profound psychosocial impacts across the globe. In this analysis of the International COVID-19 Awareness and Response Evaluation (iCARE) survey study, we comparatively investigated the psychosocial effects of COVID-19 on individuals with cancer and people with other chronic illness. iCARE study respondents were divided into two groups on the basis of self-reported health status: (1) active/current cancer (with or without any other chronic condition: heart disease, lung disease, hypertension, diabetes, severe obesity, immunity disease, and depressive or anxiety disorder); and (2) other chronic condition, but not cancer. Linear regressions were conducted to evaluate the associations between health status and outcomes. Worldwide, 18,154 iCARE study respondents (mean age, 50.8 years) from 175 countries were included in the analysis. Among them, 3.8% (n = 677) identified as having active/current cancer and 96.2% (n = 17,477) identified as having other chronic condition. Multivariate analyses showed significant associations between having cancer and declined mental (β = .364; P < .0001) and physical (β = .317; P < .0001) health since the start of the COVID-19 pandemic, relative to those with other chronic illness. Moreover, individuals with cancer demonstrated a higher likelihood of reporting maladaptive coping mechanisms such as increased alcohol use (β = .457; P < .0001) and financial hardships such as not paying rent/mortgage (β = .476; P < .0001), compared with people with other chronic illness. Individuals with cancer worldwide tended to have worse psychosocial and financial challenges during the COVID-19 pandemic, compared with other chronic disease populations. Clinicians need to be aware of the importance of attending to the specific mental health needs of individuals with cancer during and after COVID-19-related restrictions.


Autoradiographic visualization of angiotensin-converting enzyme in rat brain with [3H]captopril: localization to a striatonigral pathway.
We have visualized angiotensin-converting enzyme (ACE; dipeptidyl carboxypeptidase, peptidylpeptide hydrolase, EC 220.127.116.11) in rat brain by in vitro [3H]captopril autoradiography. [3H]Captopril binding to brain slices displays a high affinity (Kd = 1.8 X 10(-9) M) and a pharmacological profile similar to that of ACE activity. Very high densities of [3H]captopril binding were found in the choroid plexus and the subfornical organ. High densities were present in the caudate putamen and substantia nigra, zona reticulata. Moderate levels were found in the entopeduncular nucleus, globus pallidus, and median eminence of the hypothalamus. Lower levels were detectable in the supraoptic and paraventricular nuclei of the hypothalamus, the medial habenula, the median preoptic area, and the locus coeruleus. Injection of ibotenic acid or colchicine into the caudate putamen decreased [3H]captopril-associated autoradiographic grains by 85% in the ipsilateral caudate putamen and by greater than 50% in the ipsilateral substantia nigra. Thus, ACE in the substantia nigra is located on presynaptic terminals of axons originating from the caudate putamen, and ACE in the caudate putamen is situated in neuronal perikarya or at the terminals of striatal interneurons. The lack of effect of similar injections into the substantia nigra confirmed that the caudate putamen injections did not cause trans-synaptic changes. The presence of [3H]captopril binding is consistent with an ACE-mediated production of angiotensin II in some brain regions. Although [3H]captopril autoradiography reveals ACE in a striatonigral pathway, there is no evidence for angiotensin II involvement in such a neuronal pathway.


Folic Acid Protected Neural Cells Against Aluminum-Maltolate-Induced Apoptosis by Preventing miR-19 Downregulation.
Aluminum (Al)-induced apoptosis is considered as the major cause of its neurotoxicity. Folic acid possesses neuroprotective function by preventing neural cell apoptosis. microRNAs (miRNAs) are important regulators of gene expression participating in cellular processes. As a key component of the miR-17-92 cluster, miR-19 is implicated in regulating apoptotic process, while its role in the neuroprotective effect of folic acid has not been investigated. The present study aimed to investigate the potential involvement and function of miR-19 in the protective action of folic acid against Al-induced neural cell apoptosis. Human SH-SY5Y cells were treated with Al-maltolate (Al-malt) in the presence or absence of folic acid. Results showed that Al-malt-induced apoptosis of SH-SY5Y cells was effectively prevented by folic acid. Al-malt suppressed the expression of miR-19a/19b, along with alterations of miR-19 related apoptotic proteins including PTEN, p-AKT, p53, Bax, Bcl-2, caspase 9 and caspase 3; and these effects were ameliorated by folic acid. miR-19 inhibitor alone induced apoptosis of SH-SY5Y cells. Combination treatment of folic acid and miR-19 inhibitor diminished the neuroprotective effect of folic acid. These findings demonstrated that folic acid protected neuronal cells against Al-malt-induced apoptosis by preventing the downregulation of miR-19 and modulation of miR-19 related downstream PTEN/AKT/p53 pathway.


Stronger Association between Nucleus Accumbens Density and Body Mass Index in Low-Income and African American Children.
The nucleus accumbens' (NAc) size, function, and density influence individuals' body mass index (BMI). However, little is known about racial and socioeconomic status (SES) differences in the role of NAc density as a predictor of childhood BMI. We used the Adolescent Brain Cognitive Development (ABCD) data to investigate racial and SES differences in the effect of NAc density on childhood BMI. This cross-sectional study included 9497 children between ages 9 and 10. Mixed-effects regression models were used to analyze the data. The predictor variable was NAc density measured using diffusion MRI (dMRI). The outcome variable was BMI, operationalized as a continuous variable. Covariates included sex, age, ethnicity, family structure, and parental education. Race (White, African American, Asian, and Other/mixed) and household income (< 50k, 50-100 k, and 100+ k) were the moderators. High NAc diffusion tension (density) was predictive of higher BMI, net of covariates. However, the positive association between NAc density and BMI was stronger in African Americans than in White, and in low-income than in high-income children. Our findings suggest that although high NAc has implications for children's BMI, this effect varies across racial and SES groups. More research should be performed on the role of obesogenic environments in altering the effect of NAc on childhood BMI.


Unassisted detection of depression by GPs: who is most likely to be misclassified?
Meta-analyses indicate 50% of cases of depression are not detected by GPs. It is important to examine patient and GP characteristics associated with misclassification so that systems can be improved to increase accurate detection and optimal management for groups at risk of depression. To examine patient and GP characteristics associated with GP misclassification of depression for patients classified by the Patient Health Questionnaire-9 as depressed. A cross-sectional study within general practices in two states of Australia. GPs completed a one-page paper and pencil survey indicating whether they thought each patient was clinically depressed. Patients completed a computer tablet survey while waiting for their appointment to provide demographic information and indicate depression status. Chi-square analyses were used to determine whether patient and GP characteristics were associated with a false-negative and false-positive result. The probability of misclassification was modelled using Generalized Estimating Equations to account for clustering of patients. Fifty GPs from 12 practices participated. GPs completed surveys for 1880 patients. Younger patients aged 25-44, and those with a health care card were less likely to have a false-negative assessment. Patients with 0-3 GP visits in the past 12 months, and those with private health insurance were less likely to have a false-positive assessment. GPs who worked five sessions or fewer per week were more likely to make false-positive assessments.


RNA export factor RAE1 contributes to NUP98-HOXA9-mediated leukemogenesis.
Chromosomal translocations involving chimeric fusions of the nucleoporin NUP98 protein have often been described in acute myelogenous leukemia (AML). All the fusion proteins have an identical NUP98 N terminus, which contains the GLEBS motif for interaction with the mRNA export factor RAE1 and FG repeats that associate with the transcription factors HDAC1 and p300. It is virtually unknown whether these interaction partners affect leukemogenesis. We previously showed that RAE1 depletion caused aneuploidy, which enhanced tumorigenesis. We speculated that RAE1 may also be directly involved in NUP98 fusion-mediated leukemogenesis. We show here that RNA interference (RNAi)-mediated knockdown of NUP98 caused severe chromosome segregation defects and disrupted RAE1 but not HDAC1 expression and localization. Next, we performed rescue experiments to confirm that the RAE1-NUP98 complex orchestrates proper chromosome segregation. Interestingly, we found diverse behaviors of NUP98 and the leukemogenic fusion protein NUP98-HOXA9 throughout the cell cycle. Strikingly, in NUP98-HOXA9-transfected cells, RAE1 protein were reduced and mis-localized. Our cellular interpretations were further confirmed by NUP98-HOXA9 transgenic mice and the NUP98-HOXA9 AML patient. These data suggest that RAE1 orchestrates NUP98-mediated leukemogenesis and raise the possibility that targeting this negative feedback loop may provide a new strategy for the therapy of aggressive leukemias.


[Diagnostic image (4)(Bronchoalveolar lavage fluid)].
Two apparently healthy male volunteers, 28 and 32 years old, were examined before participation in a drug trial. Bronchial fluid was obtained. The lavage fluid from the second volunteer who had smoked 20 cigarettes a day for the past 16 years showed marked brownish discolouration, as compared to the transparent fluid of the non smoking first volunteer.


Nitrogen and food production: proteins for human diets.
Nitrogen was the most commonly yield-limiting nutrient in all pre-industrial agricultures. Only the Haber-Bosch synthesis of ammonia broke this barrier. The rising dependence on nitrogenous fertilizers, which represents the largest human interference in the biospheric N cycle, has two different roles. In affluent nations it helps to produce excess of food in general, and of animal foods in particular, and it boosts agricultural exports. But for at least a third of humanity in the world's most populous countries the use of N fertilizers makes the difference between malnutrition and adequate diet. Our understanding of human N (protein) needs has undergone many revisions and although some uncertainties still remain it is clear that average protein intakes are excessive in rich countries and inadequate for hundreds of millions of people in Asia, Africa, and Latin America. More dietary protein will be needed to eliminate these disparities but the future global use of N fertilizers can be moderated not just by better agronomic practices but also by higher feeding efficiencies and by gradual changes of prevailing diets. As a result, it could be possible to supply adequate nutrition to the world's growing population without any massive increases of N inputs.


Hybrid Microfluidic Platform for Multifactorial Analysis Based on Electrical Impedance, Refractometry, Optical Absorption and Fluorescence.
This paper describes the development of a novel microfluidic platform for multifactorial analysis integrating four label-free detection methods: electrical impedance, refractometry, optical absorption and fluorescence. We present the rationale for the design and the details of the microfabrication of this multifactorial hybrid microfluidic chip. The structure of the platform consists of a three-dimensionally patterned polydimethylsiloxane top part attached to a bottom SU-8 epoxy-based negative photoresist part, where microelectrodes and optical fibers are incorporated to enable impedance and optical analysis. As a proof of concept, the chip functions have been tested and explored, enabling a diversity of applications: (i) impedance-based identification of the size of micro beads, as well as counting and distinguishing of erythrocytes by their volume or membrane properties; (ii) simultaneous determination of the refractive index and optical absorption properties of solutions; and (iii) fluorescence-based bead counting.


Proteoglycans of the human intervertebral disc. Electrophoretic heterogeneity of the aggregating proteoglycans of the nucleus pulposus.
Nuclei pulposi were dissected from lumbar discs of radiologically normal human spines of cadavers aged 17, 20 and 21 years. Proteoglycans were extracted with 4 M guanidine hydrochloride (dissociative conditions) with proteinase inhibitors and isolated as A1 fractions by associative density-gradient centrifugation. Aggregating and non-aggregating proteoglycans were separated by Sepharose 2B chromatography. Both aggregating and non-aggregating proteoglycans contained a keratan sulphate-rich region as isolated by chondroitinase/trypsin/chymotrypsin digestion and Sepharose CL-6B chromatography. Agarose/acrylamide-gel electrophoresis of individual fractions of a Bio-Gel A-50m dissociative-column separation of the aggregating proteoglycans revealed two, well-separated bands: S and F, the slower and faster migrating bands respectively. The non-aggregating proteoglycan fractions were eluted under associative conditions (0.5 M-sodium acetate, pH 6.8) and migrated as a single band in the electrophoretic system. The gel-electrophoretic heterogeneity of the aggregating proteoglycans was still evident after hydroxylamine fragmentation and removal of the hyaluronate-binding portion of the molecule. Dissociative density-gradient centrifugation of the aggregating proteoglycans partially separated the Band-S proteoglycans from the Band-F population. Subsequent dissociative chromatography of the high-buoyant-density Band F proteoglycans permitted discrimination of this band into two gel-electrophoresis-distinguishable populations (Bands F-1 and F-2). Enzyme-linked immunosorbent assays with a monoclonal antibody that recognized keratan sulphate demonstrated that the D1 fraction containing the Band F-1 proteoglycans was enriched in keratan sulphate compared with the total aggregating or non-aggregating pool of proteoglycans. The proteoglycans of young adult nucleus pulposus could then be ascribed to one of four structurally and/or electrophoretically distinct populations: (1) the non-aggregating population, which comprised about 70% of the total extractable proteoglycans; (2) the aggregating pool, comprising: (a) Band F-1 proteoglycans, which had a relatively large hydrodynamic size, uronate/protein weight ratio, were enriched in keratan sulphate and had a high buoyant density; (b) Band S proteoglycans, which migrated slower in agarose/acrylamide gels, had a smaller hydrodynamic size, lower buoyant density and a lower uronate/protein ratio than the Band F-1 population; (c) Band F-2 proteoglycans, which were lower in buoyant density, smaller in hydrodynamic size and slightly faster in electrophoretic mobility than the Band F-1 proteoglycans.


Protocolization of Analgesia and Sedation Through Smart Technology in Intensive Care: Improving Patient Safety.
The risk of medication errors in intensive care units is high, primarily in the drug administration phase. Management of high-alert medications within intensive care units in the study institution varied widely. The aim of this quality improvement project was to protocolize and centralize the management of high-alert medications in acute care settings and to implement smart intravenous infusion pump technology in intensive care units. The project was conducted in 4 phases: (1) protocolization and standardization of intravenous mixtures, (2) centralization of intravenous mixture preparation in the Pharmacy Department, (3) programming of the smart pumps, and (4) dissemination and staged implementation of intravenous mixture protocols. Smart pumps (Alaris, CareFusion) were used to deliver the medicines, and the manufacturer's software (Alaris Guardrails, CareFusion) was used to analyze data regarding adherence to the drug library and the number of programming errors detected. Morphine, remifentanil, fentanyl, midazolam, dexmedetomidine, and propofol were included. After implementation of the smart pumps, 3283 infusions were started; of these, 2198 were programmed through the drug library, indicating 67% compliance with the safety software. The pumps intercepted 398 infusion-related programming errors that led to cancellation or reprogramming of drug infusions. Protocolization and centralization of the preparation of high-alert sedative and analgesic medications for critically ill patients and the administration of these drugs using smart pump technology decrease variability of clinical practice and intercept potentially serious medication errors.


Low uptake of antiretroviral therapy after admission with human immunodeficiency virus and tuberculosis in KwaZulu-Natal, South Africa.
A prospective cohort study was conducted among human immunodeficiency virus (HIV) infected in-patients with tuberculosis (TB) or other opportunistic infections (OIs) in South Africa to estimate subsequent antiretroviral therapy (ART) uptake and survival. Logistic regression modeling explored associations between baseline characteristics and starting ART, and ART exposure-adjusted incidence of death was estimated over 6 months of follow-up. Among 49 participants enrolled, median CD4 cell count at hospital discharge was 42 cells/microl and the most common presenting OIs were TB (76%), Pneumocystis pneumonia (8%), chronic diarrhea (8%), cryptococcal meningitis (6%), and Toxoplasma gondii (4%). By 6 months, only 20 (45%) patients had initiated ART, and four (8%) were lost to follow-up. ART uptake was independently associated with previous use of traditional medicine (OR 7.2, 95%CI 1.4-55.1) and with less advanced HIV infection (baseline CD4 count per 50 cells/microl increase OR 1.4, 95%CI 0.9-2.2). A total of 14 (31%) patients died before initiating ART; the monthly incidence of death did not decrease over the 6-month interval. The high mortality observed within the 6 months following hospitalization with TB or other acute OIs indicate that mechanisms are needed to expedite ART for patients after an acquired immune-deficiency syndrome defining illness.


Predicting the future of excitation energy transfer in light-harvesting complex with artificial intelligence-based quantum dynamics.
Exploring excitation energy transfer (EET) in light-harvesting complexes (LHCs) is essential for understanding the natural processes and design of highly-efficient photovoltaic devices. LHCs are open systems, where quantum effects may play a crucial role for almost perfect utilization of solar energy. Simulation of energy transfer with inclusion of quantum effects can be done within the framework of dissipative quantum dynamics (QD), which are computationally expensive. Thus, artificial intelligence (AI) offers itself as a tool for reducing the computational cost. Here we suggest AI-QD approach using AI to directly predict QD as a function of time and other parameters such as temperature, reorganization energy, etc., completely circumventing the need of recursive step-wise dynamics propagation in contrast to the traditional QD and alternative, recursive AI-based QD approaches. Our trajectory-learning AI-QD approach is able to predict the correct asymptotic behavior of QD at infinite time. We demonstrate AI-QD on seven-sites Fenna-Matthews-Olson (FMO) complex.


Valorization of the Red Seaweed Gracilaria gracilis Through a Biorefinery Approach.
The interest in seaweeds as an abundant feedstock to obtain valuable and multitarget bioactive ingredients is continuously growing. In this work, we explore the potential of Gracilaria gracilis, an edible red seaweed cultivated worldwide for its commercial interest as a source of agar and other ingredients for cosmetic, pharmacological, food, and feed applications. G. gracilis growth conditions were optimized through vegetative propagation and sporulation while manipulating the physicochemical conditions to achieve a large biomass stock. Green extraction methodologies with ethanol and water were performed over the seaweed biomass. The bioactive potential of extracts was assessed through a set of in vitro assays concerning their cytotoxicity, antioxidant, and antimicrobial properties. Additionally, dried seaweed biomass was incorporated into pasta formulations to increase food's nutritional value. Pigments extracted from G. gracilis have also been incorporated into yogurt as a natural colorant, and their stability was evaluated. Both products were submitted to the appreciation of a semi-trained sensorial panel aiming to achieve the best final formulation before reaching the market. Results support the versatility of G. gracilis whether it is applied as a whole biomass, extracts and/or pigments. Through implementing several optimized protocols, this work allows the development of products with the potential to profit the food, cosmetic, and aquaculture markets, promoting environmental sustainability and a blue circular economy. Moreover, and in line with a biorefinery approach, the residual seaweed biomass will be used as biostimulant for plant growth or converted to carbon materials to be used in water purification of the in-house aquaculture systems of MARE-Polytechnic of Leiria, Portugal.


[Perceptions and attitudes among parents and guardians on the use of pediatric medicines and their cariogenic and erosive potential].
The study assesses the perceptions of parents/guardians on the use of pediatric medicines and their cariogenic/erosive potential, interviewing them on the relationship between the use of pediatric medicines and cavities/dental erosion, as well as oral hygiene procedures after the medication. Among the interviewed parents/guardians (n=86), 24 (27.9%) mentioned the sour taste of pediatric medication, 51 (59.3%) the bitter taste, and 56 (65.1%) considered them sweet, although only 16 (28.6%) of these parents/guardians brushed their children's teeth after the medication (Fisher's Exact; p>0.05). Moreover, only 18 (20.9%) of the parents/guardians performed oral hygiene practices after the medication, of whom 8 (44.4%) received previous instructions (Fisher's Exact; p<0.05). The association between taking medications and dental caries or structural dental defects was mentioned by 61 (70.9%) of the parents/guardians, with 24 (39.3%) of them justifying this association through the presence of sugar in the formulations. Although parents/guardians consider medicines to be sweet substances and relate their use to dental caries and tooth structure flaws, most of them do not brush their children's teeth after taking medications and are unable to establish a clear cause/effect relationship for this association.


[Bacterial meningitis in Denmark 2002-2003. A nation-wide laboratory-based registration].
Notification of bacterial meningitis (BM) is likely to be incomplete, and a recent Danish study indicated that unbalanced notification may bias expected aetiology of BM. Therefore the Danish Bacterial Meningitis Group initiated a national registration of culture-positive BM. Laboratory data on all bacterial isolates from cerebrospinal fluid deemed to be clinically relevant were identified on a national basis during 2002 and 2003. Bacterial findings were compared to cases notified during the same period. A total of 404 BM cases were identified. The distribution of bacterial species differed from the national notification data during the same period especially with respect to Staphylococcus aureus (6.7% vs. 0.6%), Escherichia coli (3.5% vs. 0%) and non-Streptococcus pneumoniae streptococci (10% vs. 1.9%). The overall notification-rate was 66%, and was below 20% for S. aureus, E. coli, Enterococcus faecalis, non-S. pneumoniae streptococci and for 13 cases of "other bacteria". Sensitivity to antibiotics in the BM cases was as expected for Northern Europe. Reduced sensitivity to penicillin was found in 2/202 S. pneumoniae, in 2/10 Listeria monocytogenes, and 21/27 S. aureus were penicillin resistant. E. coli was resistant to ampicillin in 5 of 13 cases and to gentamicin in 1 of 11 cases. A suboptimal notification rate with an unbalanced species distribution was found. Laboratory based data thus constitute an improved basis for future recommendations for empirical treatment of BM. A continued national collaboration may promote the development of quality indicators for diagnosis and initial treatment of BM.


A systematic review on the causes of the transmission and control measures of outbreaks in long-term care facilities: Back to basics of infection control.
The unique characteristics of long-term care facilities (LTCFs) including host factors and living conditions contribute to the spread of contagious pathogens. Control measures are essential to interrupt the transmission and to manage outbreaks effectively. The aim of this systematic review was to verify the causes and problems contributing to transmission and to identify control measures during outbreaks in LTCFs. Four electronic databases were searched for articles published from 2007 to 2018. Articles written in English reporting outbreaks in LTCFs were included. The quality of the studies was assessed using the risk-of-bias assessment tool for nonrandomized studies. A total of 37 studies were included in the qualitative synthesis. The most commonly reported single pathogen was influenza virus, followed by group A streptococcus (GAS). Of the studies that identified the cause, about half of them noted outbreaks transmitted via person-to-person. Suboptimal infection control practice including inadequate decontamination and poor hand hygiene was the most frequently raised issue propagating transmission. Especially, lapses in specific care procedures were linked with outbreaks of GAS and hepatitis B and C viruses. About 60% of the included studies reported affected cases among staff, but only a few studies implemented work restriction during outbreaks. This review indicates that the violation of basic infection control practice could be a major role in introducing and facilitating the spread of contagious diseases in LTCFs. It shows the need to promote compliance with basic practices of infection control to prevent outbreaks in LTCFs.


Evaluation of Patients with Cleft Lip and Palate in Southeastern Anatolian Region.
Our study included patients with cleft lip and palate who applied to Gaziantep University Dentistry Faculty Orthodontics Department between 2010 and 2014. The aim of this study was to reveal the characteristics of these patients with demographic values. In total, 118 patients were included in this study. The patients were pursued by phone calls, and the study was conducted with only 64 patients. The demographic map was created with a statistical expert as a questionnaire. It was thought that the reason for not being able to include the remaining 54 patients was a change in phone number, designation, or others. One patient who was to be interviewed died because of heart failure. The results revealed that the place of birth of 18.75% patients was a metropolitan region that of 54.68% was a province, that of 21.87% was a district, and that of 4.69% was a village. The majority of patients (93.25%) had a low family income level. The average number of children was 3.28, and readership was 1.47. The rate of working mothers was 1.56%. It was concluded that the cleft lip and palate patient group had lower economic and sociocultural levels. For this purpose, raising awareness of parents and providing financial support to families are required because of the long-term treatment process. It was thought that the long-term treatment process of these patients in a single-unit authority with the guidance of universities would be appropriate for better patient follow-up and inventory. Otherwise, as seen in this study, contact cannot be established after a while and some problems can arise with regard to follow-up. Lack of communication can lead to the formation of more difficult patients with growth and development.


Genetic determinants for methotrexate response in juvenile idiopathic arthritis.
Juvenile idiopathic arthritis (JIAs) is the most common chronic rheumatic disease of childhood and is an important cause of disability. The folic acid analog methotrexate is the first choice disease-modifying anti-rheumatic drug in this disease, however, 35-45% of patients fail to respond. Molecular elements, such as variants in genes of pharmacological relevance, influencing response to methotrexate in JIA, would be important to individualize treatment strategies. Several studies have evaluated the effects of candidate genetic variants in the complex pathway of genes involved in methotrexate pharmacodynamics and pharmacokinetics, however, results are still contrasting and no definitive genetic marker of methotrexate response useful for the clinician to tailor therapy of children with JIA has been identified. Recently, genome-wide approaches have been applied, identifying new potential biological processes involved in methotrexate response in JIA such as TGF-beta signaling and calcium channels. If these genomic results are properly validated and integrated with innovative analyses comprising deep sequencing, epigenetics, and pharmacokinetics, they will greatly contribute to personalize therapy with methotrexate in children with JIA.


Training to improve awareness of disabilities in clients with unilateral neglect.
Awareness of disabilities is known to be a central problem of rehabilitation among clients with large right cerebrovascular lesions and unilateral neglect. The aim of this study was to evaluate the effect of an intervention program focused on improving the awareness of disabilities in four participants with unilateral neglect. The intervention program developed for this study was based on the assumption that awareness of disabilities is a prerequisite for being able to learn and use compensatory techniques in the performance of activities of daily living (ADL). The study followed a single-case experimental ABA design. The Assessment of Awareness of Disability was used to measure awareness of disabilities; the Assessment of Motor and Process Skills was used to measure ADL ability; and neuropsychological tests were used to assess unilateral neglect and sustained attention. The intervention program used meaningful and purposeful occupations as therapeutic change agents to improve awareness of disabilities. Awareness of disabilities and ADL ability improved in all four participants; unilateral neglect decreased in three participants; and sustained attention improved in two participants. The preliminary findings indicate that training to improve awareness of disabilities might improve the ability to learn the use of compensatory techniques in the performance of ADL in clients with unilateral neglect. The effects of the intervention strategy need to be evaluated further in future research.


Extraversion personality, perceived health and activity participation among community-dwelling aging adults in Hong Kong.
Activity participation is essential to the wellbeing of aging adults. Divergent levels of activity participation within aging populations have been explained from diverse perspectives, but the interaction effects of key determinants, such as personality and health, are often ignored. This study examines the effects of extravert personality on aging adults' activity levels by addressing its interaction with perceived physical health and mental health. A sample of 304 adults aged 50 and older was selected using systematic sampling from participants of an institute for promoting active aging at a university in Hong Kong in 2017. Data on socio-demographic characteristics, perceived physical and mental health, extraversion personality traits, and level of activity participation were collected using a telephone survey. Most participants (46.7%) reported moderate activity levels and over a quarter (26.6%) reported high or low activity levels. Multi-nominal logistic regression analyses show that extraversion was associated with an increased likelihood of reporting moderate (OR = 1.85, p = .036) but not high (p > .05) activity levels when adjusted for perceived physical and mental health and socio-demographics, with low activity levels being the constant comparison. Meanwhile, extraversion predicted both moderate (OR = 3.84, p = .014) and high (OR = 5.06, p = .032) activity levels for participants with poor or average perceived mental health. However, the interaction effects of extraversion with perceived physical health or mental health were not significant in predicting either moderate or high activity levels (p > .05). The implications for enhancing activity participation among aging adults are discussed in view of both personality and perceived health status.


Organising knowledge to prevent global health crises: a comparative analysis of pandemic preparedness indicators.
Preparing for the possibility of a global pandemic presents a transnational organisational challenge: to assemble and coordinate knowledge over institutionally diverse countries with high fidelity. The COVID-19 pandemic has laid these problems bare. This article examines the construction of the three main cross-national indicators of pandemic preparedness: a database with self-reported data by governments, external evaluations organised by the WHO and a global ranking known as the Global Health Security Index. Each of these presents a different model of collecting evidence and organising knowledge: the collation of self-reports by national authorities; the coordination of evaluation by an epistemic community authorised by an intergovernmental organisation and on the basis of a strict template; and the cobbling together of different sources into a common indicator by a transnational multi-stakeholder initiative. We posit that these models represent different ways of creating knowledge to inform policy choices, and each has different forms of potential bias. In turn, this shapes how policymakers understand what is 'best practice' and appropriate policy in pandemic preparedness.


Stimulus-Driven Attention, Threat Bias, and Sad Bias in Youth with a History of an Anxiety Disorder or Depression.
Attention biases towards threatening and sad stimuli are associated with pediatric anxiety and depression, respectively. The basic cognitive mechanisms associated with attention biases in youth, however, remain unclear. Here, we tested the hypothesis that threat bias (selective attention for threatening versus neutral stimuli) but not sad bias relies on stimulus-driven attention. We collected measures of stimulus-driven attention, threat bias, sad bias, and current clinical symptoms in youth with a history of an anxiety disorder and/or depression (ANX/DEP; n = 40) as well as healthy controls (HC; n = 33). Stimulus-driven attention was measured with a non-emotional spatial orienting task, while threat bias and sad bias were measured at a short time interval (150 ms) with a spatial orienting task using emotional faces and at a longer time interval (500 ms) using a dot-probe task. In ANX/DEP but not HC, early attention bias towards threat was negatively correlated with later attention bias to threat, suggesting that early threat vigilance was associated with later threat avoidance. Across all subjects, stimulus-driven orienting was not correlated with early threat bias but was negatively correlated with later threat bias, indicating that rapid stimulus-driven orienting is linked to later threat avoidance. No parallel relationships were detected for sad bias. Current symptoms of depression but not anxiety were related to decreased stimulus-driven attention. Together, these results are consistent with the hypothesis that threat bias but not sad bias relies on stimulus-driven attention. These results inform the design of attention bias modification programs that aim to reverse threat biases and reduce symptoms associated with pediatric anxiety and depression.


Implications of antibiotics use during the COVID-19 pandemic: present and future.
COVID-19 is caused by the SARS-CoV-2 virus, which has infected more than 4 million people with 278 892 deaths worldwide as of 11 May 2020. This disease, which can manifest as a severe respiratory infection, has been declared as a public health emergency of international concern and is being treated with a variety of antivirals, antibiotics and antifungals. This article highlights the administration of antimicrobials in COVID-19 patients worldwide, during the 2019-20 pandemic. It is imperative to be aware of the unreported amounts of antibiotics that have been administered worldwide in just a few months and a marked increase in antimicrobial resistance should therefore be expected. Due to the lack of data about antimicrobial use during this pandemic, the global impact on the emergence of new antimicrobial resistance is as yet unknown. This issue must be at the forefront of public health policymaking and planning in order that we are prepared for the potentially severe consequences for human and animal health and the environment.


Factors related to maternal mortality rate in covid-19 patients - a cross-sectional study from an Indonesian covid-19 referral hospital.
The covid-19 pandemic may cause severe clinical manifestations in a vulnerable population, such as pregnant women. Based on Indonesian Obstetrics and Gynecology Association (POGI), the number of maternal deaths due to covid-19 from April 2020 to April 2021 reached 3% and increased to 9% since the delta variant of covid-19 emerged. This research was expected to identify factors that are related to the mortality rate of pregnant women with covid-19. This was a cross-sectional study using secondary data collected from June 2020 to August 2021. The study was conducted in Universitas Indonesia Hospital, a national covid-19 referral hospital. Patient characteristics, pregnancy profile, comorbidities, laboratory results, chest X-ray examination, treatment options, and the severity of symptoms were evaluated. In addition, bivariate data analysis was carried out using the SPSS device. Out of 114 research subjects, seven patients (6.1%) died, and 107 patients (93.9%) survived. The risk of mortality was significantly (P < 0.05) related to patients' age, duration of hospitalization, gestational age, severity rate of covid-19, the level of hemoglobin, leukocyte count, platelet count, lymphocytes, the levels of D-dimer, C-reactive protein, transaminase enzymes, urea, creatinine, eGFR, sodium, potassium, and procalcitonin. In addition, significant differences (P < 0.05) related to maternal mortality rate were also shown in the presence of comorbidities (type 2 diabetes, congestive heart failure, coronary artery disease/acute coronary syndrome, and urinary tract infection), and the use of steroids and tocilizumab. Various factors significantly related to the mortality rate of pregnant women with covid-19. This study may become the basis for a further study with a larger number of subjects, adjustment of assessment and management of covid-19 infected pregnant women, thus hopefully reducing the risk of mortality in pregnant women with covid-19.


Dramatic repositioning of c-Myb to different promoters during the cell cycle observed by combining cell sorting with chromatin immunoprecipitation.
The c-Myb transcription factor is a critical regulator of proliferation and stem cell differentiation, and mutated alleles of c-Myb are oncogenic, but little is known about changes in c-Myb activity during the cell cycle. To map the association of c-Myb with specific target genes during the cell cycle, we developed a novel Fix-Sort-ChIP approach, in which asynchronously growing cells were fixed with formaldehyde, stained with Hoechst 33342 and separated into different cell cycle fractions by flow sorting, then processed for chromatin immunoprecipitation (ChIP) assays. We found that c-Myb actively repositions, binding to some genes only in specific cell cycle phases. In addition, the specificity of c-Myb is dramatically different in small subpopulations of cells, for example cells in the G2/M phase of the cell cycle, than in the bulk population. The repositioning of c-Myb during the cell cycle is not due to changes in its expression and also occurs with ectopically expressed, epitope-tagged versions of c-Myb. The repositioning occurs in established cell lines, in primary human CD34+ hematopoietic progenitors and in primary human acute myeloid leukemia cells. The combination of fixation, sorting and ChIP analysis sheds new light on the dynamic nature of gene regulation during the cell cycle and provides a new type of tool for the analysis of gene regulation in small subsets of cells, such as cells in a specific phase of the cell cycle.


Texture and Structure Measurements and Analyses for Evaluation of Fish and Fillet Freshness Quality: A Review.
Recently, food safety and quality have become critical issues of great concern throughout the world. Fish is one of the most vulnerable and perishable aquatic products. The evaluation of fish and fillet freshness is therefore very significant in research and development for providing premium and supreme quality for human health and acceptance by consumers, as well as for international trade. The texture and structure of fish muscle are important freshness quality attributes that depend on several parameters such as hardness, cohesiveness, springiness, chewiness, resilience, and adhesiveness, as well as the internal cross-linking of connective tissue and the detachment of fibers. This review aims to present recent advances of texture and structure measurements and analyses, including sensory evaluation and instrumental methods, for indicating and evaluating fish freshness quality. Factors affecting these measurements are detailed and correlations between texture and structure are discussed. Moreover, the limitations and challenges of fish texture and structure measurements are described and some viewpoints about current work and future trends are also presented.


Comparative study of number and distribution of IgG cells in oral lichen planus and oral lichenoid lesions.
Oral lichen planus is a common mucocutaneous disorder with unknown etiology. While current data suggest that oral lichen planus is a cell-mediated disease, differential diagnosis of this disease and oral lichenoid lesions is very problematic, both clinically and histopathologically. This study aimed to compare immunohistochemical features of these similar diseases. This was a descriptive-analytic study in which formalin-fixed and paraffin-embedded tissue sections of 30 oral lichen planus and 30 oral lichenoid lesions were immunohistochemically analyzed for number and distribution of IgG(+) cells. A standard biotin-streptavidin procedure after antigen retrieval was used. Data were analyzed in SPSS software using Mann-Whitney U test. There were some significant differences in distribution of IgG (+)cells among different locations in oral lichen planus and also in oral lichenoid lesions separately; but the differences between distribution of IgG(+) cells between the two groups of oral lichen planus and oral lichenoid lesions were not significant. There was no significant difference in number and distribution of IgG(+) cells between the two groups. So, this study can suggest that location of IgG is similar in samples of oral lichen planus and oral lichenoid lesions and consequently, this marker cannot help us differentiate them from each other. Other markers can be analyzed in further studies in order to find an appropriate distinguisher between the two lesions.


AltHapAlignR: improved accuracy of RNA-seq analyses through the use of alternative haplotypes.
Reliance on mapping to a single reference haplotype currently limits accurate estimation of allele or haplotype-specific expression using RNA-sequencing, notably in highly polymorphic regions such as the major histocompatibility complex. We present AltHapAlignR, a method incorporating alternate reference haplotypes to generate gene- and haplotype-level estimates of transcript abundance for any genomic region where such information is available. We validate using simulated and experimental data to quantify input allelic ratios for major histocompatibility complex haplotypes, demonstrating significantly improved correlation with ground truth estimates of gene counts compared to standard single reference mapping. We apply AltHapAlignR to RNA-seq data from 462 individuals, showing how significant underestimation of expression of the majority of classical human leukocyte antigen genes using conventional mapping can be corrected using AltHapAlignR to allow more accurate quantification of gene expression for individual alleles and haplotypes. Source code freely available at https://github.com/jknightlab/AltHapAlignR. Supplementary data are available at Bioinformatics online.


Comparative Analysis of Muscle Synergy between Patients with Shoulder Impingement Syndrome and Healthy Controls: A Preliminary Study.
Shoulder impingement syndrome can be caused by a muscle imbalance around the shoulder joint. It would be beneficial for therapy to implement rehabilitation exercises based on intermuscular coordination in order to achieve muscle balance. This study presents the muscle synergy characteristics of patients with shoulder impingement syndrome, which can be used to develop rehabilitation exercises. During pick and place task, the muscle synergy obtained from eight shoulder muscles in patients and healthy subjects was compared. The experimental results revealed that patients have low contributed muscle synergy structures for the serratus anterior and infraspinatus.Clinical relevance- This proposes that patients with shoulder impingement syndrome may have abnormal muscle synergy structure which can be used for assessment.


[Caries prevalence and dental hygiene in a group of school-age children].
The results of a cross-sectional study of the prevalence of caries in a sample of 3,237 children aged 5 to 21 are described in relation to some objectives of the WHO program. The prevalence of caries is high in the sample: 1. 31.5% of the children aged 5 to 6 are free of caries (according to the WHO objectives, 50% of the children aged 5 to 6 should have no caries), 2. the mean number of decayed, missing and filled permanent teeth (DMF/T index) in children aged 11 to 12 is 6.6 (The DMF/T should be less than or equal to 3). The frequency of toothbrushing is distributed as follows: 23% of the children brush their teeth once a week or less, 46% brush their teeth once a day, and 31% brush their teeth twice a day or more. The purpose and the frequency of the consultations at the dentist's office have been examined: 15% of the children never consult the dentist, 44% consult the dentist in case of toothache, and 41% go to the dentist's office for screening or control. Only 3.2% of the children use regularly and appropriately fluoride tablets. Since the variance of the results is large, it is difficult to distinguish explaining variables or determinants. However, the data indicate that social groups show differences in DMF indices as well as in the behaviors associated with dental hygiene and prevention.


Strongly Coupled Redox-Linked Conformational Switching at the Active Site of the Non-Heme Iron-Dependent Dioxygenase, TauD.
2-Oxoglutarate (2OG)-dependent dioxygenases catalyze C-H activation while performing a wide range of chemical transformations. In contrast to their heme analogues, non-heme iron centers afford greater structural flexibility with important implications for their diverse catalytic mechanisms. We characterize an in situ structural model of the putative transient ferric intermediate of 2OG:taurine dioxygenase (TauD) by using a combination of spectroelectrochemical and semiempirical computational methods, demonstrating that the Fe(III/II) transition involves a substantial, fully reversible, redox-linked conformational change at the active site. This rearrangement alters the apparent redox potential of the active site between -127 mV for reduction of the ferric state and +171 mV for oxidation of the ferrous state of the 2OG-Fe-TauD complex. Structural perturbations exhibit limited sensitivity to mediator concentrations and potential pulse duration. Similar changes were observed in the Fe-TauD and taurine-2OG-Fe-TauD complexes, thus attributing the reorganization to the protein moiety rather than the cosubstrates. Redox-difference infrared spectra indicate a reorganization of the protein backbone in addition to the involvement of carboxylate and histidine ligands. Quantitative modeling of the transient redox response using two alternative reaction schemes across a variety of experimental conditions strongly supports the proposal for intrinsic protein reorganization as the origin of the experimental observations.


Structural and serological evidence for a novel mechanism of antigenic variation in foot-and-mouth disease virus.
Changes resulting in altered antigenic properties of viruses nearly always occur on their surface and have been attributed to the substitution of residues directly involved in binding antibody. To investigate the mechanism of antigenic variation in foot-and-mouth disease virus (FMDV), variants that escape neutralization by a monoclonal antibody have been compared crystallographically and serologically with parental virus. FMDVs form one of the four genera of the Picornaviridae. The unenveloped icosahedral shell comprises 60 copies each of four structural proteins VP1-4. Representatives from each of the genera have similar overall structure, but differences in the external features. For example, human rhinovirus has a pronounced 'canyon' that is proposed to contain the cell attachment site, whereas elements of the attachment site for FMDV, which involves the G-H loop (residues 134-160) and C-terminus (200-213) of VP1, are exposed on the surface. Moreover, this G-H loop, which is a major antigenic site of FMDV, forms a prominent, highly accessible protrusion, a feature not seen in other picornaviruses. It is this loop that is perturbed in the variant viruses that we have studied. The amino acid mutations characterizing the variants are not at positions directly involved in antibody binding, but result in far-reaching perturbations of the surface structure of the virus. Thus, this virus seems to use a novel escape mechanism whereby an induced conformational change in a major antigenic loop destroys the integrity of the epitope.


How useful is the EQ-5D in assessing the impact of caring for people with Alzheimer's disease?
The impact on informal caregivers of caring for people with Alzheimer's disease (AD) dementia can be substantial, but it remains unclear which measures(s) best assess such impact. Our objective was to use data from the GERAS study to assess the ability of the EuroQol 5-dimension questionnaire (EQ-5D) to measure the impact on caregivers of caring for people with AD dementia and to examine correlations between EQ-5D and caregiver burden. GERAS was a prospective, non-interventional cohort study in community-dwelling patients with AD dementia and their informal caregivers. The EQ-5D and Zarit Burden Interview (ZBI) were used to measure health-related quality of life and caregiver burden, respectively. Resource-use data collected included caregiver time spent with the patient on activities of daily living (ADL). Spearman correlations were computed between EQ-5D scores, ZBI scores, and time spent on instrumental ADL (T-IADL) at baseline, 18 months, and for 18-month change scores. T-IADL and ZBI change scores were summarized by EQ-5D domain change category (better/stable/worse). At baseline, 1495 caregivers had mean EQ-5D index scores of 0.86, 0.85, and 0.82, and ZBI total scores of 24.6, 29.4, and 34.1 for patients with mild, moderate, and moderately severe/severe AD dementia, respectively. Change in T-IADL showed a stronger correlation with change in ZBI (0.12; P < 0.001) than with change in EQ-5D index score (0.02; P = 0.546) although both correlations were very weak. Worsening within EQ-5D domains was associated with increases in ZBI scores, although 68%-90% of caregivers remained stable within each EQ-5D domain. There was no clear pattern for change in T-IADL by change in EQ-5D domain. EQ-5D may not be the optimum measure of the impact of caring for people with AD dementia due to its focus on physical health. Alternative measures need further investigation.


Expression of individual members of a tRNA(Gly)1 multigene family in vivo follows the same pattern as in vitro.
Individual members of a tRNA(Gly)1 multigene family from Bombyx mori are transcribed to different levels in vitro in homologous nuclear extracts but their transcription status in vivo is not known. Two sets of tRNA(Gly)1 belonging to the extreme groups of highly transcribed and barely transcribed copies have been examined for their expression patterns in vivo in B. mori-derived cell lines following transfection. We have developed a sensitive and reliable method for directly quantifying the transcription levels of transfecting tRNA genes without relying on the biological activity of the transcript. The strategy involved the insertion of synthetic oligodeoxyribonucleotide sequences into the coding region of the transfecting gene and monitoring the transcripts in an RNase protection assay using an antisense probe that clearly distinguished them from the endogenous tRNAs. The oligonucleotide insertion did not significantly affect the transcriptional status of the genes, even though the distance between the A and B boxes was enhanced by 10-15 nt. In vivo also the transcription of tRNA(Gly)1 reached very high levels, whereas the transcripts arising from tRNA(Gly)1-6:7 accounted for only 2-5% of the former, closely resembling the transcription patterns in vitro. These individual gene copies having identical coding sequences and consequently the same internal conserved regions, differed only in their flanking sequences which modulate their transcription levels.


On meta- and mega-analyses for gene-environment interactions.
Gene-by-environment (G × E) interactions are important in explaining the missing heritability and understanding the causation of complex diseases, but a single, moderately sized study often has limited statistical power to detect such interactions. With the increasing need for integrating data and reporting results from multiple collaborative studies or sites, debate over choice between mega- versus meta-analysis continues. In principle, data from different sites can be integrated at the individual level into a "mega" data set, which can be fit by a joint "mega-analysis." Alternatively, analyses can be done at each site, and results across sites can be combined through a "meta-analysis" procedure without integrating individual level data across sites. Although mega-analysis has been advocated in several recent initiatives, meta-analysis has the advantages of simplicity and feasibility, and has recently led to several important findings in identifying main genetic effects. In this paper, we conducted empirical and simulation studies, using data from a G × E study of lung cancer, to compare the mega- and meta-analyses in four commonly used G × E analyses under the scenario that the number of studies is small and sample sizes of individual studies are relatively large. We compared the two data integration approaches in the context of fixed effect models and random effects models separately. Our investigations provide valuable insights in understanding the differences between mega- and meta-analyses in practice of combining small number of studies in identifying G × E interactions.


What Is Caring in Nursing?: Sorting Out Humanistic and Christian Perspectives.
Historically, some propose that organized nursing was founded on biblical principles and the caring behaviors and characteristics of Christ, and that caring originates with God. More recently, perceptions of caring have shifted to a humanistic, postmodern worldview, where the source of caring is within the person. This integrative literature review synthesized caring literature from humanistic, postmodern, and biblical, theological worldviews. Findings reveal that 95% of research and 85% of nonresearch publications were written from the humanistic perspective. The results lay a foundation for an alternative middle range theory on caring from a biblical worldview for nurses holding Christian beliefs.


Gaze Behavior in Basketball Free Throws Developed in Constant and Variable Practice.
There are a limited number of studies focusing on the mechanisms explaining why variable practice gives an advantage in a novel situation and constant practice in performance in trained conditions. We hypothesized that this may be due to the different gaze behavior that is developed under different conditions. Twenty participants, randomly assigned to two different groups, practiced basketball free throws for three consecutive days, performing 100 throws per day. The constant group (n = 10) practiced at a free throw distance (4.57 m) only. The variable practice group (n = 10) randomly performed 20 shots per five throw distances (3.35, 3.96, 4.57, 5.18, and 5.79 m) on each day, also accumulating 100 shots per day. We analyzed the total gaze fixation duration, a number of fixations, and the average fixation duration on a basketball rim in a pretest and posttest at the 4.57 m distance. We computed a linear mixed model with test (pretest-posttest), group (constant-variable), and test × group interaction in order to analyze the total fixation duration and number of fixations. The average fixation duration was analyzed with a repeated measure two-way ANOVA, with practice conditions as a between-participants factor and test type as a within-participants factor. We found that the total fixation duration increased significantly in the posttest, regardless of the practice conditions (p < 0.001, effect size = 0.504). The number of fixations also increased significantly in the posttest (p = 0.037, effect size = 0.246). The average fixation duration increased in both groups; however, insignificantly. We also did not find any significant differences between groups. Our results suggest that variable and constant practice conditions may lead to the development of similar gaze behavior.


After critical care: patient support after critical care. A mixed method longitudinal study using email interviews and questionnaires.
To explore experiences and needs over time, of patients discharged from ICU using the Intensive Care Experience (ICE-q) questionnaire, Hospital Anxiety and Depression Scale (HADS) and EuroQoL (EQ-5D), associated clinical predictors (APACHE II, TISS, Length of stay, RIKER scores) and in-depth email interviewing. A mixed-method, longitudinal study of patients with >48hour ICU stays at 2 weeks, 6 months, 12 months using the ICE-q, HADS, EQ-5D triangulated with clinical predictors, including age, gender, length of stay (ICU and hospital), APACHE II and TISS. In-depth qualitative email interviews were completed at 1 month and 6 months. Grounded Theory analysis was applied to interview data and data were triangulated with questionnaire and clinical data. Data was collected from January 2010 to March 2012 from 77 participants. Both mean EQ-5D visual analogue scale, utility scores and HADS scores improved from 2 weeks to 6 months, (p=<0.001; p=<0.001), but between 6 and 12 months, no change was found in data from either questionnaire, suggesting improvements level off. These variations were reflected in qualitative data themes: rehabilitation/recovery in the context of chronic illness; impact of critical care; emotional and psychological needs (including sub-themes of: information needs and relocation anxiety). The overarching, core theme related to adjustment of normality. Patient recovery in this population appears to be shaped by ongoing illness and treatment. Email interviews offer a convenient method of gaining in-depth interview data and could be used as part of ICU follow-up.


Adaptation of tonic accommodation reduces accommodative lag and is masked in darkness.
Tonic aftereffects of accommodation were investigated under three open loop conditions; Maxwellian view (pin hole pupil), empty field, and darkness. Duration of accommodative aftereffects, following 5 sec of adaptation, ranged within several seconds for all three open loop conditions. After adapting for 1 min to a 2-D stimulus, the duration of accommodative aftereffects increased by approximately an order of magnitude for the Maxwellian view and empty field conditions; however, they increased modestly for the dark condition. Tonic aftereffects of accommodation were also stimulated by convergence with base out prism or divergence with base in prism while the accommodative loop was opened. The ratio of open-loop aftereffect amplitude/closed-loop accommodative response to lenses ranged from 0.95 to 0.48, and it became smaller when the accommodative stimulus approached the amplitude of accommodation. The rapid decay of tonic aftereffects of accommodation in darkness was reversable when a visible stimulus was presented while the accommodative loop was opened by Maxwellian view or empty field. Hence, darkness only masked aftereffects that were manifest primarily when there was light stimulation of the retina. The amplitudes of accommodative aftereffects were reciprocally related to the amplitude of the lag of accommodation. During a 2-min adaptation period, accommodative response increased gradually for 30-60 sec as it approached the amplitude of the accommodative stimulus. This resulted in a reduction of accommodative error or lag. Opening the accommodative loop before or after the lag was reduced resulted in a small or large aftereffect of accommodation respectively. Accommodative aftereffects appear to be related to accommodative lag as prism adaptation is related to fixation disparity.


Evaluation of resource allocation and supply-demand balance in clinical practice with high-cost technologies.
Japan has one of the highest numbers of high-cost medical devices installed relative to its population. While evaluations of the distribution of these devices traditionally involve simple population-based assessments, an indicator that includes the demand of these devices would more accurately reflect the situation. The purpose of this study was to develop an indicator of the supply-demand balance of such devices, using examples of magnetic resonance imaging scanners (MRI) and extracorporeal shockwave lithotripters (ESWL), and to investigate the relationship between this indicator, personnel distribution statuses and operating statuses at the prefectural level. Using data from nation-wide surveys and claims data from 16 hospitals, we developed an indicator based on the ratio of the supplied number of device units to the number of device units in demand for MRI and ESWL. The latter value was based on patient volume and utilization proportion. Correlation analyses were conducted between the supply-demand balances of these devices, personal distribution and operating statuses. Comparisons between our indicator and conventional population-based indicators revealed that 15% and 30% of prefectures were at risk of underestimating the availability of MRI and ESWL, respectively. The numbers of specialist personnel/device units showed significant, negative correlations with our indicators in both devices. Utilization-based analyses of health care resource placement and utilization status provide a more accurate indication than simple population-based assessments, and can assist decision makers in reviewing gaps between health policy and management. Such an indicator therefore has the potential to be a tool in helping to improve the efficiency of the allocation and placement of such devices.


A multi-signal fluorescent probe for simultaneously distinguishing and sequentially sensing cysteine/homocysteine, glutathione, and hydrogen sulfide in living cells.
Biothiols, which have a close network of generation and metabolic pathways among them, are essential reactive sulfur species (RSS) in the cells and play vital roles in human physiology. However, biothiols possess highly similar chemical structures and properties, resulting in it being an enormous challenge to simultaneously discriminate them from each other. Herein, we develop a unique fluorescent probe (HMN) for not only simultaneously distinguishing Cys/Hcy, GSH, and H2S from each other, but also sequentially sensing Cys/Hcy/GSH and H2S using a multi-channel fluorescence mode for the first time. When responding to the respective biothiols, the robust probe exhibits multiple sets of fluorescence signals at three distinct emission bands (blue-green-red). The new probe can also sense H2S at different concentration levels with changes of fluorescence at the blue and red emission bands. In addition, the novel probe HMN is able to discriminate and sequentially sense biothiols in biological environments via three-color fluorescence imaging. We expect that the development of the robust probe HMN will provide a powerful strategy to design fluorescent probes for the discrimination and sequential detection of biothiols, and offer a promising tool for exploring the interrelated roles of biothiols in various physiological and pathological conditions.


Carboxydobrachium pacificum gen. nov., sp. nov., a new anaerobic, thermophilic, CO-utilizing marine bacterium from Okinawa Trough.
A new anaerobic, thermophilic, CO-utilizing marine bacterium, strain JMT, was isolated from a submarine hot vent in Okinawa Trough. Cells of strain JMT were non-motile thin straight rods, sometimes branching, with a cell wall of the Gram-positive type, surrounded with an S-layer. Chains of three to five cells were often observed. The isolate grew chemolithotrophically on CO, producing equimolar quantities of H2 and CO2 (according to the equation CO+H2O-->CO2+H2) and organotrophically on peptone, yeast extract, starch, cellobiose, glucose, galactose, fructose and pyruvate, producing H2, acetate and CO2. Growth was observed from 50 to 80 degrees C with an optimum at 70 degrees C. The optimum pH was 6.8-7.1. The optimum concentration of sea salts in the medium was 20.5-25.5 g l(-1). The generation time under optimal conditions was 7.1 h. The DNA G+C content was 33 mol %. Growth of isolate JMT was not inhibited by penicillin, but ampicillin, streptomycin, kanamycin and neomycin completely inhibited growth. The results of 16S rDNA sequence analysis revealed that strain JMT belongs to the Thermoanaerobacter phylogenetic group within the Bacillus-Clostridium subphylum of Gram-positive bacteria but represents a separate branch of this group. On the basis of morphological and physiological features and phylogenetic data, this isolate should be assigned to a new genus, for which the name Carboxydobrachium is proposed. The type species is Carboxydobrachium pacificum; the type strain is JMT (= DSM 12653T).


Cardiotoxicity and cardiovascular disease risk assessment for patients receiving breast cancer treatment.
Cardiotoxicity from anticancer therapy affects heart function and structure. Cardiotoxicity can also lead to accelerated development of chronic diseases, especially in the presence of risk factors. This study aimed to develop and pilot a combined cardiovascular disease and cardiotoxicity risk assessment questionnaire to quantify the potential extent of risk factors in breast cancer patients prior to treatment. The questionnaire underwent content and face validity evaluation by an expert panel followed by pilot testing in a sample of breast cancer patients (n = 36). Questionnaires were self-administered while attending chemotherapy clinic, in the presence of a research assistant. Mean age of participants was 54.8 years (range 36-72 years). Participants reported CVD risk factors including diabetes 2.8%, hypertension 19.8%, hypercholesterolaemia 11% and sleep apnoea 5%. Lifestyle risk factors, included not eating the recommended serves of vegetables (100%) or fruit (78%) per day; smoking (13%) and regularly consuming alcohol (75%). Twenty five percent reported being physically inactive, 61%, overweight or obese, 24%, little or no social support and 30% recorded high to very high psychological distress. Participants were highly (75%) reluctant to undertake lifestyle changes; i.e. changing alcohol consumption; dietary habits; good emotional/mental health strategies; improving physical activity; quitting smoking; learning about heart-health and weight loss. This study is an important step towards prevention and management of treatment-associated cardiotoxicity after breast cancer diagnosis. We recommend that our questionnaire is providing important data that should be included in cancer registries so that researchers can establish the relationship between CVD risk profile and cardiotoxicity outcomes and that this study revealed important teaching opportunities that could be used to examine the impact on health literacy and help patients better understand the consequences of cancer treatment.


Kidney transplant candidates' views of the transplant allocation system.
The point system used to distribute scarce transplantable kidneys places great emphasis on antigen matching. This contributes to increased waiting times for African Americans, who have a disproportionate share of rare antigens. We conducted a pilot study to explore the understanding and attitudes of kidney transplant candidates toward the way the transplant allocation system trades off between antigen matching and waiting time. We performed semi-structured interviews of a convenience sample of 33 patients awaiting transplants in Philadelphia and its surrounding suburbs. Patients had a number of misconceptions about the transplant allocation system. Many incorrectly thought, for example, that quality of life and financial status influence which patients on the waiting list receive available organs. Despite these and other misconceptions, the majority of patients thought the allocation system was fair. However, many African Americans thought the system was biased against them because of their race. After hearing about how the transplant system factors antigen matching and waiting time into organ allocation, the majority of subjects still felt the system was fair. After hearing that the emphasis on antigen matching causes African Americans to wait twice as long as whites, a larger number of subjects thought the system was unfair. Nevertheless, few thought the system should be changed. Even African American patients who felt the system was unfair still approved of the emphasis on antigen matching out of a desire to have a successful kidney transplant. We found that most of the interviewed patients awaiting kidney transplant thought the system should continue to emphasize antigen matching. Although attitudes toward the allocation system differed by race, with African American patients more suspicious of the system, the importance patients placed on antigen matching did not appear to differ by race.


Sequence dependence of the folding of collagen-like peptides. Single amino acids affect the rate of triple-helix nucleation.
The refolding of thermally denatured model collagen-like peptides was studied for a set of 21 guest triplets embedded in a common host framework: acetyl-(Gly-Pro-Hyp)3-Gly-Xaa-Yaa-(Gly-Pro-Hyp)4-Gly-Gly-amide. The results show a strong dependence of the folding rate on the identity of the guest Gly-Xaa-Yaa triplet, with the half-times for refolding varying from 6 to 110 min (concentration = 1 mg/ml). All triplets of the form Gly-Xaa-Hyp promoted rapid folding, with the rate only marginally dependent on the residue in the Xaa position. In contrast, triplets of the form Gly-Pro-Yaa and Gly-Xaa-Yaa were slower and showed a wide range of half-times, varying with the identity of the residues in the triplet. At low concentrations, the folding can be described by third-order kinetics, suggesting nucleation is rate-limiting. Data on the relative nucleation ability of different Gly-Xaa-Yaa triplets support the favorable nature of imino acids, the importance of hydroxyproline, the varying effects of the same residue in the Xaa position versus the Yaa position, and the difficulties encountered when leucine or aspartic acid are in the Yaa position. Information on the relative propensities of different tripeptide sequences to promote nucleation of the triple-helix in peptides will aid in identification of nucleation sites in collagen sequences.


'Toning' up hypotonia assessment: A proposal and critique.
Clinical assessment of hypotonia is challenging due to the subjective nature of the initial clinical evaluation. This poses dilemmas for practitioners in gaining accuracy, given that the presentation of hypotonia can be either a non-threatening or malevolent sign. The research question posed was how clinical assessment can be improved, given the current contentions expressed in the scientific literature. This paper describes the development and critique of a clinical algorithm to aid the assessment of hypotonia. An initial exploratory sequential phase, consisting of a systematic review, a survey amongst clinicians and a Delphi process, assisted in the development of the algorithm, which is presented within the framework of the International Classification of Functioning, Disability and Health. The ensuing critique followed a qualitative emergent-systematic focus group design with a purposive sample of 59 clinicians. Data were analysed using semantical content analysis and are presented thematically with analytical considerations. This study culminated in the development of an evidence-based clinical algorithm for practice. The qualitative critique of the algorithm considered aspects such as inadequacies, misconceptions and omissions; strengths; clinical use; resource implications; and recommendations. The first prototype and critique of a clinical algorithm to assist the clinical assessment of hypotonia in children has been described. Barriers highlighted include aspects related to knowledge gaps of clinicians, issues around user-friendliness and formatting concerns. Strengths identified by the critique included aspects related to the evidence-based nature of the criteria within the algorithm, the suitability of the algorithm in being merged or extending current practice, the potential of the algorithm in aiding more accurate decision-making, the suitability of the algorithm across age groups and the logical flow. These findings provide a starting point towards ascertaining the clinical utility of the algorithm as an essential step towards evidence-based praxis.


Pulled Flowprobe for Ambient Liquid Extraction-Based High Spatial Resolution Mass Spectrometry Imaging with Enhanced Sensitivity and Stability.
Ambient liquid extraction techniques enable direct mass spectrometry imaging (MSI) under ambient conditions with minimal sample preparation. However, currently an integrated probe for ambient liquid extraction-based MSI with high spatial resolution, high sensitivity, and stability is still lacking. In this work, we developed a new integrated probe made of pulled coaxial capillaries, named pulled flowprobe, and compared it with the previously reported single-probe. Mass transfer kinetics in probes was first investigated. The extraction kinetic curves during probe sampling indicate a narrower and higher peak shape for the pulled flowprobe than single-probe. Computational fluid dynamics analysis reveals that in the pulled flowprobe flow velocities are lower in liquid microjunction and higher in the transferring channels, resulting in higher extraction efficiencies and reduced band diffusion compared with single-probe and other probes with a similar flow route. Results of ambient liquid extraction-based MSI of lipids in rat cerebrum show that signals of low-abundance lipids were 2-5 times higher via a pulled flowprobe than via a single-probe, and 26 more lipid species were detected on brain tissue via a pulled flowprobe than via a single-probe. The stability of MSI with the pulled flowprobe was found to be higher than that with single-probe (averaged relative standard deviation = 18% vs 80%) by imaging a lab-made uniform ink coating. Moreover, in the pulled flowprobe, no retraction of the inner capillary from outer capillary is optimal for both sensitivity and stability. The spatial resolution of the pulled flowprobe (30-40 μm) was measured to be higher than that of a comparable size single-probe by calculation with the "80-20" rule. Finally, the new pulled flowprobe was applied to high-resolution MSI of lipids in the hippocampus, and localization of several lipids to the specific cell layers in the hippocampus region was observed. Thus, this work provides an alternative easily fabricated sampling probe with enhanced sensitivity, stability, and spatial resolution, promoting the use of ambient liquid extraction-based MSI in biological and clinical research.


Identification of a 64 kDa heparan sulphate proteoglycan core protein from human lung fibroblast plasma membranes with a monoclonal antibody.
Human lung fibroblasts produce heparan sulphate proteoglycans (HSPG) that are associated with the plasma membrane. A monoclonal-antibody (Mab)-secreting hybridoma, S1, was produced by fusion of SP 2/0-AG 14 mouse myeloma cells with spleen cells from mice immunized with partially purified cellular HSPG fractions. The HSPG character of the material carrying the epitope recognized by Mab S1 was demonstrated by: (i) the co-purification of the S1 epitope with the membrane HSPG of human lung fibroblasts; (ii) the decrease in size of the material carrying the S1 epitope upon treatment with heparinase or heparitinase, and the resistance of this material to heparinase treatment after N-desulphation. The S1 epitope appears to be part of the core protein, since it was destroyed by proteinase treatment and by disulphide-bond reduction, but not by treatments that depolymerize the glycosaminoglycan chains and N-linked oligosaccharide chains. Polyacrylamide-gel electrophoresis of non-reduced heparitinase-digested membrane HSPG followed by Western blotting and immunostaining with Mab S1 revealed a single band with apparent molecular mass of 64 kDa. Membrane proteoglycans isolated from detergent extracts or from 4 M-guanidinium chloride extracts of the cells yielded similar results. Additional digestion with N-glycanase lowered the apparent molecular mass of the immunoreactive material to 56 kDa, suggesting that the core protein also carries N-linked oligosaccharides. Fractionation of 125I-labelled membrane HSPG by immuno-affinity chromatography on immobilized Mab S1, followed by heparitinase digestion and polyacrylamide-gel electrophoresis of the bound material, yielded a single labelled band with apparent molecular mass 64 kDa. Treatment with dithiothreitol caused a slight increase in apparent molecular mass, suggesting that the core protein of this membrane proteoglycan of a single subunit containing (an) intrachain disulphide bond(s).


Raloxifene improves endothelial dysfunction in hypertension by reduced oxidative stress and enhanced nitric oxide production.
It has not been completely clarified whether selective estrogen receptor modulators (SERMs) such as raloxifene exert vasoprotective effects similar to those of estrogens. To investigate vascular effects of raloxifene, male spontaneously hypertensive rats were treated for 10 weeks with either raloxifene (10 mg x kg(-1) x d(-1)) or vehicle. Raloxifene improved endothelium-dependent vasodilatation but had no effect on either endothelium-independent vasorelaxation or phenylephrine-induced vasoconstriction. Raloxifene treatment increased the release of NO from the vessel wall by enhanced expression and activity of endothelial NO synthase. Blood pressure reduction after bradykinin infusion was more pronounced in animals treated with SERMs. The production of superoxide in intact aortic segments was decreased by raloxifene treatment. Administration of raloxifene had no effect on the expression of the essential NAD(P)H oxidase subunits p22phox and nox1 in the vasculature but reduced the activity and expression of vascular membrane-bound rac1, a GTPase required for the activation of the NAD(P)H oxidase. Finally, blood pressure levels were significantly decreased in spontaneously hypertensive rats treated with raloxifene. All SERM effects were also detected in healthy age-matched Wistar rats. In cultured rat aortic vascular smooth muscle cells, raloxifene inhibited angiotensin II-induced reactive oxygen species production dependent on estrogen receptor activation. Raloxifene treatment improves hypertension-induced endothelial dysfunction by increased bioavailability of NO. This is achieved by an increased activity of endothelial NO synthase and by an estrogen receptor-dependent reduction in release of reactive oxygen species from vascular cells. These vascular effects cause a profound blood pressure reduction and lead to decreased vascular damage in male spontaneously hypertensive rats.


Label- and amplification-free electrochemical detection of bacterial ribosomal RNA.
Current approaches to molecular diagnostics rely heavily on PCR amplification and optical detection methods which have restrictions when applied to point of care (POC) applications. Herein we describe the development of a label-free and amplification-free method of pathogen detection applied to Escherichia coli which overcomes the bottleneck of complex sample preparation and has the potential to be implemented as a rapid, cost effective test suitable for point of care use. Ribosomal RNA is naturally amplified in bacterial cells, which makes it a promising target for sensitive detection without the necessity for prior in vitro amplification. Using fluorescent microarray methods with rRNA targets from a range of pathogens, an optimal probe was selected from a pool of probe candidates identified in silico. The specificity of probes was investigated on DNA microarray using fluorescently labeled 16S rRNA target. The probe yielding highest specificity performance was evaluated in terms of sensitivity and a LOD of 20 pM was achieved on fluorescent glass microarray. This probe was transferred to an EIS end point format and specificity which correlated to microarray data was demonstrated. Excellent sensitivity was facilitated by the use of uncharged PNA probes and large 16S rRNA target and investigations resulted in an LOD of 50 pM. An alternative kinetic EIS assay format was demonstrated with which rRNA could be detected in a species specific manner within 10-40min at room temperature without wash steps.


Using a noninfectious MVM surrogate for assessing viral clearance during downstream process development.
Viral contamination is an inherent risk during the manufacture of biopharmaceuticals. As such, biopharmaceutical companies must demonstrate the viral clearance efficacy of their downstream process steps prior to clinical trials and commercial approval. This is accomplished through expensive and logistically challenging spiking studies, which utilize live mammalian viruses. These hurdles deter companies from analyzing viral clearance during process development and characterization. We utilized a noninfectious minute virus of mice-mock virus particle (MVM-MVP) as a surrogate spiking agent during small scale viral filtration (VF) and anion exchange chromatography (AEX) studies. For VF experiments, in-process mAb material was spiked and processed through Asahi Kasei P15, P20, P35, and BioEX nanofilters. Across each filter type, flux decay profiles and log reduction values (LRVs) were nearly identical for either particle. For AEX experiments, loads were conditioned with various amounts of sodium chloride (9, 20, 23, and 41 mS/cm), spiked with either particle and processed through a Q-SFF packed column. LRV results met our expectations of predicting MVM removal.


Determinants of cesarean delivery in the US: a lifecourse approach.
This study takes a lifecourse approach to understanding the factors contributing to delivery methods in the US by identifying preconception and pregnancy-related determinants of medically indicated and non-medically indicated cesarean section (C-section) deliveries. Data are from the Early Childhood Longitudinal Study-Birth Cohort, a nationally representative, population-based survey of women delivering a live baby in 2001 (n = 9,350). Three delivery methods were examined: (1) vaginal delivery (reference); (2) medically indicated C-section; and (3) non-medically indicated C-sections. Using multinomial logistic regression, we examined the role of sociodemographics, health, healthcare, stressful life events, pregnancy complications, and history of C-section on the odds of medically indicated and non-medically indicated C-sections, compared to vaginal delivery. 74.2 % of women had a vaginal delivery, 11.6 % had a non-medically indicated C-section, and 14.2 % had a medically indicated C-section. Multivariable analyses revealed that prior C-section was the strongest predictor of both medically indicated and non-medically indicated C-sections. However, we found salient differences between the risk factors for indicated and non-indicated C-sections. Surgical deliveries continue to occur at a high rate in the US despite evidence that they increase the risk for morbidity and mortality among women and their children. Reducing the number of non-medically indicated C-sections is warranted to lower the short- and long-term risks for deleterious health outcomes for women and their babies across the lifecourse. Healthcare providers should address the risk factors for medically indicated C-sections to optimize low-risk delivery methods and improve the survival, health, and well-being of children and their mothers.


WITHDRAWN: Mefloquine for preventing malaria in non-immune adult travellers.
Mefloquine is commonly prescribed to prevent malaria in travellers, and has replaced other drugs because Plasmodium falciparum is commonly resistant to them. However, mefloquine may be associated with neuropsychiatric harmful effects. To assess the effects of mefloquine in adult travellers compared to other regimens in relation to episodes of malaria, withdrawal from prophylaxis, and adverse events. We searched the Cochrane Infectious Diseases Group specialized trials register (September 2002), The Cochrane Central Register of Controlled Trials (The Cochrane Library Issue 3, 2002), MEDLINE (1966 to September 2002), EMBASE (1980 to September 2002), LILACS (September 2002), Science Citation Index (1981 to September 2002), and bibliographies in retrieved papers and standard textbooks. We contacted researchers in the subject of malaria chemoprophylaxis, and pharmaceutical companies. Randomised trials comparing mefloquine with other standard prophylaxis or placebo in non-immune adult travellers, and in non-travelling volunteers. For adverse events, any published case reports were collected. We independently assessed trial quality and extracted data. Adverse events from observational studies were categorised by the study type. We also contacted study authors. We included 10 trials involving 2750 non-immune adult participants. Five of these were field trials, and of these all were in mainly male soldiers. One trial comparing mefloquine with placebo showed mefloquine prevented malaria episodes in an area of drug resistance (Peto odds ratio 0.04, 95% confidence interval 0.02 to 0.08). Withdrawals in the mefloquine group were consistently higher in four placebo controlled trials (odds ratio 3.56, 95% confidence interval 1.67 to 7.60). In five trials comparing mefloquine with other chemoprophylaxis, no difference in tolerability was detected. We found 516 published case reports of mefloquine adverse effects. 63 per cent of these published reports involved tourists and business travellers. There were four fatalities attributed to mefloquine. Mefloquine prevents malaria, but has adverse effects that limit its acceptability . There is evidence from non-randomised studies that mefloquine has potentially harmful effects in tourists and business travellers, and its use needs to be carefully balanced against this. Trials of comparative effects of antimalarial prophylaxis should include episodes of malaria and withdrawal from prophylaxis as outcomes.


In utero radiation-induced apoptosis and p53 gene expression in the developing rat brain.
This study addressed the question of the role of the p53 gene in prenatal low-dose radiation-induced apoptosis in the neuroepithelium, in an effort to elucidate molecular mechanisms involved in the extreme radiosensitivity of the developing brain. Pregnant Wistar rats were exposed to a single dose of 10, 20 or 40 cGy of X-rays on day 15 or 17 of gestation. Animals were sacrificed 4 or 24h after exposure. Apoptosis was studied by gel electrophoresis of isolated DNA and in situ by the TUNEL reaction. Expression of the p53 gene was studied by immunocytochemistry and Western analysis, as well as Northern analysis, for the detection of the protein and mRNA respectively. In utero low-dose irradiation led to apoptosis and an increase of p53 gene expression in the developing rat brain. Apoptotic as well as p53 immunopositive cells were detected among proliferating, migratory and post-mitotic neurones in the developing neuroepithelium following prenatal irradiation, even after only l0 cGy. In addition to the p53 protein, p53 mRNA brain levels were also increased following prenatal irradiation. Low-dose prenatal irradiation of the developing brain led to p53 induction and cell death by apoptosis.


Tumor-Informed Approach Improved ctDNA Detection Rate in Resected Pancreatic Cancer.
Pancreatic cancer is one of the cancers with very poor prognosis; there is an urgent need to identify novel biomarkers to improve its clinical outcomes. Circulating tumor DNA (ctDNA) from liquid biopsy has arisen as a promising biomarker for cancer detection and surveillance. However, it is known that the ctDNA detection rate in resected pancreatic cancer is low compared with other types of cancer. In this study, we collected paired tumor and plasma samples from 145 pancreatic cancer patients. Plasma samples were collected from 71 patients of treatment-naïve status and from 74 patients after neoadjuvant therapy (NAT). Genomic profiling of tumor DNA and plasma samples was conducted using targeted next-generation sequencing (NGS). Somatic mutations were detected in 85% (123/145) of tumors. ctDNA was detected in 39% (28/71) and 31% (23/74) of treatment-naïve and after-NAT groups, respectively, without referring to the information of tumor profiles. With a tumor-informed approach (TIA), ctDNA detection rate improved to 56% (40/71) and 36% (27/74) in treatment-naïve and after-NAT groups, respectively, with the detection rate significantly improved (p = 0.0165) among the treatment-naïve group compared to the after-NAT group. Cases who had detectable plasma ctDNA concordant to the corresponding tumor showed significantly shorter recurrence-free survival (RFS) (p = 0.0010). We demonstrated that TIA improves ctDNA detection rate in pancreatic cancer, and that ctDNA could be a potential prognostic biomarker for recurrence risk prediction.


Structure of the newly found green turtle egg-white ribonuclease.
Marine green turtle (Chelonia mydas) egg-white ribonuclease (GTRNase) was crystallized from 1.1 M ammonium sulfate pH 5.5 and 30% glycerol using the sitting-drop vapour-diffusion method. The structure of GTRNase has been solved at 1.60 A resolution by the molecular-replacement technique using a model based on the structure of RNase 5 (murine angiogenin) from Mus musculus (46% identity). The crystal belonged to the monoclinic space group C2, with unit-cell parameters a = 86.271, b = 34.174, c = 39.738 A, alpha = 90, beta = 102, gamma = 90 degrees . GTRNase consists of three helices and seven beta-strands and displays the alpha+beta folding topology typical of a member of the RNase A superfamily. Superposition of the C(alpha) coordinates of GTRNase and RNase A superfamily members indicates that the overall structure is highly similar to that of angiogenin or RNase 5 from M. musculus (PDB code 2bwl) and RNase A from Bos taurus (PDB code 2blz), with root-mean-square deviations of 3.9 and 2.0 A, respectively. The catalytic residues are conserved with respect to the RNase A superfamily. The three disulfide bridges observed in the reptilian enzymes are conserved in GTRNase, while one further disulfide bond is required for the structural stability of mammalian RNases. GTRNase is expressed in egg white and the fact that its sequence has the highest similarity to that of snapping turtle pancreatic RNase suggests that the GTRNase secreted from oviduct cells to form egg white is probably the product of the same gene as activated in pancreatic cells.


Spending for specialized mental health treatment in The VA: 1995-2001.
The mid-1990s saw dramatic changes in mental health care in the Department of Veterans Affairs (VA), the largest provider of such care in the United States. Spending for specialized inpatient mental health care fell 21 percent from 1995 to 2001, while spending for specialized outpatient care rose 63 percent. The shift from inpatient to outpatient care was accompanied by rapid increases in outpatient medication costs. Overall, the VA reduced the average cost (per VA user) of specialized mental health care by 22 percent while it increased the number of users of these services by 35 percent.


Rational Design of Live-Attenuated Vaccines against Herpes Simplex Viruses.
Diseases caused by human herpes simplex virus types 1 and 2 (HSV-1 and HSV-2) affect millions of people worldwide and range from fatal encephalitis in neonates and herpes keratitis to orofacial and genital herpes, among other manifestations. The viruses can be shed efficiently by asymptomatic carriers, causing increased rates of infection. Viral transmission occurs through direct contact of mucosal surfaces followed by initial replication of the incoming virus in skin tissues. Subsequently, the viruses infect sensory neurons in the trigeminal and lumbosacral dorsal root ganglia, where they are primarily maintained in a transcriptionally repressed state termed "latency", which persists for the lifetime of the host. HSV DNA has also been detected in other sympathetic ganglia. Periodically, latent viruses can reactivate, causing ulcerative and often painful lesions primarily at the site of primary infection and proximal sites. In the United States, recurrent genital herpes alone accounts for more than a billion dollars in direct medical costs per year, while there are much higher costs associated with the socio-economic aspects of diseased patients, such as loss of productivity due to mental anguish. Currently, there are no effective FDA-approved vaccines for either prophylactic or therapeutic treatment of human herpes simplex infections, while several recent clinical trials have failed to achieve their endpoint goals. Historically, live-attenuated vaccines have successfully combated viral diseases, including polio, influenza, measles, and smallpox. Vaccines aimed to protect against the devastation of smallpox led to the most significant achievement in medical history: the eradication of human disease by vaccination. Recently, novel approaches toward developing safe and effective live-attenuated vaccines have demonstrated high efficacy in various preclinical models of herpetic disease. This next generation of live-attenuated vaccines has been tailored to minimize vaccine-associated side effects and promote effective and long-lasting immune responses. The ultimate goal is to prevent or reduce primary infections (prophylactic vaccines) or reduce the frequency and severity of disease associated with reactivation events (therapeutic vaccines). These vaccines' "rational" design is based on our current understanding of the immunopathogenesis of herpesviral infections that guide the development of vaccines that generate robust and protective immune responses. This review covers recent advances in the development of herpes simplex vaccines and the current state of ongoing clinical trials in pursuit of an effective vaccine against herpes simplex virus infections and associated diseases.


Robotic adherent cell injection for characterizing cell-cell communication.
Compared to robotic injection of suspended cells (e.g., embryos and oocytes), fewer attempts were made to automate the injection of adherent cells (e.g., cancer cells and cardiomyocytes) due to their smaller size, highly irregular morphology, small thickness (a few micrometers thick), and large variations in thickness across cells. This paper presents a robotic system for automated microinjection of adherent cells. The system is embedded with several new capabilities: automatically locating micropipette tips; robustly detecting the contact of micropipette tip with cell culturing surface and directly with cell membrane; and precisely compensating for accumulative positioning errors. These new capabilities make it practical to perform adherent cell microinjection truly via computer mouse clicking in front of a computer monitor, on hundreds and thousands of cells per experiment (versus a few to tens of cells as state of the art). System operation speed, success rate, and cell viability rate were quantitatively evaluated based on robotic microinjection of over 4000 cells. This paper also reports the use of the new robotic system to perform cell-cell communication studies using large sample sizes. The gap junction function in a cardiac muscle cell line (HL-1 cells), for the first time, was quantified with the system.