Accumulation of proteinase K-resistant prion protein (PrP) is restricted by the expression level of normal PrP in mice inoculated with a mouse-adapted strain of the Creutzfeldt-Jakob disease agent.
Creutzfeldt-Jakob disease (CJD) is a transmissible neurodegenerative disease of humans caused by an unidentified infectious agent, the prion. To determine whether there was an involvement of the host-encoded prion protein (PrPc) in CJD development and prion propagation, mice heterozygous (PrP+/-) or homozygous (PrP-/-) for a disrupted PrP gene were established and inoculated with the mouse-adapted CJD agent. In keeping with findings of previous studies using other lines of PrP-less mice inoculated with scrapie agents, no PrP-/- mice showed any sign of the disease for 460 days after inoculation, while all of the PrP+/- and control PrP+/+ mice developed CJD-like symptoms and died. The incubation period for PrP+/- mice, 259 +/- 27 days, was much longer than that for PrP+/+ mice, 138 +/- 12 days. Propagation of the prion was barely detectable in the brains of PrP-/- mice and was estimated to be at a level at least 4 orders of magnitude lower than that in PrP+/+ mice. These findings indicate that PrPc is necessary for both the development of the disease and propagation of the prion in the inoculated mice. The proteinase-resistant PrP (PrPres) was undetectable in the brain tissues of the inoculated PrP-/- mice, while it accumulated in the affected brains of PrP+/+ and PrP+/- mice. Interestingly, the maximum level of PrPres in the brains of PrP+/- mice was about half of the level in the similarly affected brains of PrP+/+ mice, indicating that PrPres accumulation is restricted by the level of PrPc.


Postcategorical auditory distraction in short-term memory: Insights from increased task load and task type.
Task-irrelevant speech impairs short-term serial recall appreciably. On the interference-by-process account, the processing of physical (i.e., precategorical) changes in speech yields order cues that conflict with the serial-ordering process deployed to perform the serial recall task. In this view, the postcategorical properties (e.g., phonology, meaning) of speech play no role. The present study reassessed the implications of recent demonstrations of auditory postcategorical distraction in serial recall that have been taken as support for an alternative, attentional-diversion, account of the irrelevant speech effect. Focusing on the disruptive effect of emotionally valent compared with neutral words on serial recall, we show that the distracter-valence effect is eliminated under conditions-high task-encoding load-thought to shield against attentional diversion whereas the general effect of speech (neutral words compared with quiet) remains unaffected (Experiment 1). Furthermore, the distracter-valence effect generalizes to a task that does not require the processing of serial order-the missing-item task-whereas the effect of speech per se is attenuated in this task (Experiment 2). We conclude that postcategorical auditory distraction phenomena in serial short-term memory (STM) are incidental: they are observable in such a setting but, unlike the acoustically driven irrelevant speech effect, are not integral to it. As such, the findings support a duplex-mechanism account over a unitary view of auditory distraction. (PsycINFO Database Record


Screening of colorectal cancer: present and future.
Colorectal cancer (CRC) is the third most common cancer in males and second in females, and the fourth most common cause of cancer death worldwide. Currently, about 60-70% of diagnosed cases in symptomatic patients are detected at an advanced stage of disease. Earlier stage detection through the use of screening strategies would allow for better outcomes in terms of reducing the disease burden. Areas covered: The aim of this paper is to review the current published evidence from literature which assesses the performance and effectiveness of different screening tests for the early detection of CRC. Expert commentary: Adequate screening strategies can reduce CRC incidence and mortality. In the last few decades, several tests have been proposed for CRC screening. To date, there is still insufficient evidence to identify which approach is definitively superior, and no screening strategy for CRC can therefore be defined as universally ideal. The best strategy would be the one that can be economically viable and to which the patient can adhere best to over time. The latest guidelines suggest colonoscopy every 10 years or annual fecal immuno-chemical test (FIT) for people with normal risk, while for individuals with high risk or hereditary syndromes specific recommendations are provided.


Simultaneous radiation therapy and cisplatin chemotherapy in advanced cancer of the head and neck. A pilot study at Tygerberg Hospital, Parowvallei, South Africa.
Fifty patients with advanced carcinoma of the head and neck were treated in a nonrandomized pilot study utilizing simultaneous cisplatin and radiotherapy. Treatment was delivered in two segments. The initial partial course (PC) consisted of 75-100 mg/m2 of cisplatin and concomitant radio-therapy to 20.00 Gy in 1 week, with 4-week split. Patients who tolerated therapy and experienced at least a partial response (PR) went on to complete the full course of therapy (FC), with the second phase consisting of 120 mg/m2 of cisplatin and 38.40 Gy of radiation. Total cisplatin delivered ranged between 195 and 220 mg/m2, and the total radiation dose was 58.40 Gy. Of 38 evaluable patients, 31 (82%) had at least a partial response (PR). However, at time of analysis 20 months after study, 27 (71%) patients were alive with disease or dead. Mean disease-free survival (DFS) for patients receiving FC therapy (10.05 months) was greater than that for patients who received only PC therapy (2.65 months), but the difference in overall survival (OS) was not statistically significant (11.95 FC versus 10.29 PC). Response rates and the minimal toxicity are within the range of other reported studies. Randomized studies should seek optimal time-dose schedule and establish advantages in terms of crude survival and disease-free survival.


Politics, Pandemics, and Trauma: Understanding and Addressing Latino Health Needs Through a Culturally-Informed Lens.
Latino communities in the United States (U.S.) have long endured trauma due to multiple intersecting social and political forces. New restrictive immigration policies since 2016 and the COVID-19 pandemic have each created novel stressors for Latino communities, while escalating the risk of mental health disorders and highlighting the communities' vulnerabilities. The effects of these stressors have been particularly pronounced in southeastern states, such as Tennessee and Georgia, due to their state-level anti-immigrant legislation. Yet, we lack sufficient data to understand how these factors present among Latinos seeking services. To focus attention on the specific experiences of Latino communities living in the U.S. Southeast, the authors analyzed the perspectives of 44 service providers working with these communities in the region using qualitative data collected in an online survey administered during the COVID-19 pandemic and while President Trump's exclusionary immigration policies were in force. Four themes were identified: (1) Latino communities' strengths; (2) impact of the Trump administration on Latino communities; (3) impact of COVID-19's on Latino communities; and (4) strategies to enhance service delivery in Latino communities. Results provide meaningful data to inform micro- and macro-level service delivery in two exclusionary policy states and beyond. Findings suggest future research should include other new immigrant destinations and explore perceptions of Latino community members.


Folate, vitamin B12 and homocysteine status in women of childbearing age: baseline data of folic acid wheat flour fortification in Iran.
Folic acid fortification implemented mandatorily in many countries has been associated with significant increase in blood folate concentrations and reduction in the prevalence of neural tube defects. However, there are controversial findings on the probability of vitamin B(12) deficiency being masked in the population after folic acid fortification. Baseline data on folate and vitamin B(12) status are necessary before mandatory flour fortification is implemented. To assess dietary intake of folate and vitamin B(12) and to determine blood concentrations of folate, vitamin B(12) and homocysteine in women of childbearing age as baseline data regarding folic acid fortification in Iran. A descriptive cross-sectional survey was performed with 579 healthy women as a representative sample of the Golestan province. Fasting blood samples were taken and dietary (24-hour recall), health and sociodemographic data were collected with an interview. Serum concentrations of folate and vitamin B(12) were measured with radioimmunoassay, and plasma homocysteine concentrations were assessed by high-performance liquid chromatography with fluorescence detection. Mean serum concentrations of folate and vitamin B(12) were 13.6 nmol/l (95% CI 12.8-14.4) and 194.4 pmol/l (95% CI 183.8-205.0), respectively. Inadequate serum folate levels were present in 14.3% of the women and 22.7% had serum vitamin B(12) levels below normal. Mean plasma homocysteine concentration was 12.6 micromol/l (95% CI 12.1-13.2) and hyperhomocysteinemia was observed in 38.3% of the women. Mean daily intake of folate and vitamin B(12) was 198.3 microg (95% CI 185.4-211.3) and 2.6 microg (95% CI 1.9-3.2), respectively. Folate intake from food was positively correlated with serum folate concentrations (r = 0.084, p < 0.05) and inversely correlated with plasma homocysteine concentrations (r = -0.115, p < 0.01). An insufficient vitamin B(12) as well as folate status is present in Iranian women of childbearing age. The final evaluation will be carried out 18 months after flour fortification and the results will be compared with baseline data obtained from the present study in order to show the efficacy and safety of folic acid fortification in Iran.


Association of cigarette smoking with amyotrophic lateral sclerosis.
We explored the relationship between amyotrophic lateral sclerosis (ALS) and cigarette smoking in a case-control study conducted in New England from 1993 to 1996. Recently diagnosed ALS cases (n = 109) were recruited from two major referral centers. Population controls (n = 256) were identified by random telephone screening. Data were analyzed by logistic regression. After adjusting for age, sex, region and education, ever having smoked cigarettes was associated with an increase in risk for ALS (odds ratio 1.7; 95% confidence interval 1.0-2.8). Average cigarettes smoked per day, years smoked and pack-years were all greater in cases than controls, but dose-response trends were not observed. Similar numbers of cases and controls had ever used alcohol, and only a small, nonsignificant association of drinks per month with ALS was observed. The association of cigarette smoking with ALS was not affected by adjusting for alcohol use. In contrast, the weak relationship of ALS with alcohol use was apparently due to confounding by smoking.


From Substrate to Fragments to Inhibitor Active In Vivo against Staphylococcus aureus.
Antibiotic resistance is a worldwide threat due to the decreasing supply of new antimicrobials. Novel targets and innovative strategies are urgently needed to generate pathbreaking drug compounds. NAD kinase (NADK) is essential for growth in most bacteria, as it supports critical metabolic pathways. Here, we report the discovery of a new class of antibacterials that targets bacterial NADK. We generated a series of small synthetic adenine derivatives to screen those harboring promising substituents in order to guide efficient fragment linking. This led to NKI1, a new lead compound inhibiting NADK that showed in vitro bactericidal activity against Staphylococcus aureus. In a murine model of infection, NKI1 restricted survival of the bacteria, including methicillin-resistant S. aureus. Collectively, these findings identify bacterial NADK as a potential drug target and NKI1 as a lead compound in the treatment of staphylococcal infections.


Use of gamma-glutamyl transpeptidase activity as a marker of hair cycle and anagen induction in mouse hair follicles.
gamma-glutamyl transpeptidase activity was monitored in cycling mice by histologic localization and biochemical assay. Our objective for this study is to establish the relationship between gamma-glutamyl transpeptidase activity and hair growth and to determine whether its activity can be correlated to induced hair growth. In cycling mouse skin, gamma-glutamyl transpeptidase activity is pronounced during anagen and greatly diminished during telogen. In the skin, the enzyme is present exclusively in the outer and inner root sheaths of hair follicles. gamma-glutamyl transpeptidase is limited to the follicle below the level of the sebaceous gland and is completely absent in the follicle above the sebaceous gland level. During anagen, the outer root sheath in the hypodermis is intensely positive for gamma-glutamyl transpeptidase activity whereas the hair matrix cells and dermal papillar are negative. The inner root sheath above the bulb shows distinctive membrane staining for gamma-glutamyl transpeptidase. gamma-glutamyl transpeptidase activity can be seen to vary only in cycling follicles. Inducing anagen by plucking hair shafts results in an increase in gamma-glutamyl transpeptidase activity directly correlated to hair regrowth. In a similar manner, mice were plucked and treated with a daily dose of 2% minoxidil. A slight difference in cycle lengths was seen in animals treated with minoxidil when compared to vehicle control. Minoxidil treatment may cause an early initiation of anagen, but both the minoxidil-treated skin and the vehicle-treated skin entered telogen at the same time. Together, these studies indicate that gamma-glutamyl transpeptidase is a specific marker of anagen in growing hair.


Modeling and simulation of a grand piano.
A time-domain global modeling of a grand piano is presented. The string model includes internal losses, stiffness, and geometrical nonlinearity. The hammer-string interaction is governed by a nonlinear dissipative compression force. The soundboard is modeled as a dissipative bidimensional orthotropic Reissner-Mindlin plate where the presence of ribs and bridges is treated as local heterogeneities. The coupling between strings and soundboard at the bridge allows the transmission of both transverse and longitudinal waves to the soundboard. The soundboard is coupled to the acoustic field, whereas all other parts of the structure are supposed to be perfectly rigid. The acoustic field is bounded artificially using perfectly matched layers. The discrete form of the equations is based on original energy preserving schemes. Artificial decoupling is achieved, through the use of Schur complements and Lagrange multipliers, so that each variable of the problem can be updated separately at each time step. The capability of the model is highlighted by series of simulations in the low, medium, and high register, and through comparisons with waveforms recorded on a Steinway D piano. Its ability to account for phantom partials and precursors, consecutive to string nonlinearity and inharmonicity, is particularly emphasized.


Astaxanthin Attenuates Environmental Tobacco Smoke-Induced Cognitive Deficits: A Critical Role of p38 MAPK.
Increasing evidence indicates that environmental tobacco smoke (ETS) impairs cognitive function and induces oxidative stress in the brain. Recently, astaxanthin (ATX), a marine bioactive compound, has been reported to ameliorate cognitive deficits. However, the underlying pathogenesis remains unclear. In this study, ATX administration (40 mg/kg and 80 mg/kg, oral gavage) and cigarette smoking were carried out once a day for 10 weeks to investigate whether the p38 MAPK is involved in cognitive function in response to ATX treatment in the cortex and hippocampus of ETS mice. Results indicated that ATX administration improved spatial learning and memory of ETS mice (p < 0.05 or p < 0.01). Furthermore, exposure to ATX prevented the increases in the protein levels of the p38mitogen-activated protein kinase (p38 MAPK; p < 0.05 or p < 0.01) and nuclear factor-kappa B (NF-κB p65; p < 0.05 or p < 0.01), reversed the decreases in the mRNA and protein levels of synapsin I (SYN) and postsynaptic density protein 95 (PSD-95) (all p < 0.05 or p < 0.01). Moreover, ATX significantly down-regulated the increased levels of pro-inflammatory cytokines including interleukin-6 (IL-6) and tumor necrosis factor (TNF-α) (all p < 0.05 or p < 0.01). Meanwhile, the increased level of malondialdehyde (MDA) and the decreased activities of superoxide dismutase (SOD), glutathione (GSH), and catalase (CAT) were suppressed after exposure to ATX (all p < 0.05 or p < 0.01). Also, the results of the molecular docking study of ATX into the p38 MAPK binding site revealed that its mechanism was possibly similar to that of PH797804, a p38 MAPK inhibitor. Therefore, our results indicated that the ATX might be a critical agent in protecting the brain against neuroinflammation, synaptic plasticity impairment, and oxidative stress in the cortex and hippocampus of ETS mice.


The sliding sulcus procedure: simultaneous repair of unilateral clefts of the lip and primary palate--a new technique.
During embryogenesis the clefting event unleashes a series of processes that act in concert to deform the soft tissue envelope into a "dysfunctional matrix" within which bone and cartilage subsequently form. Much of secondary cleft deformity results from failure to solve this pathologic soft tissue-bone equation at the primary repair. The sliding sulcus procedure uses the subperiosteal plane to create a centralization of the soft tissue matrix, alleviate tension, and redistribute the forces of growth in a more normal pattern. By incising at the level of the alveolus, this approach preserves the blood supply to the osteogenic cells of the buccolabial periosteum. This approach has interesting theoretical implications for the preservation of alveolar growth potential. Arch stabilization and elimination of fistula may be of long-term benefit to the orthodontist.


Regularized Modal Regression with Applications in Cognitive Impairment Prediction.
Linear regression models have been successfully used to function estimation and model selection in high-dimensional data analysis. However, most existing methods are built on least squares with the mean square error (MSE) criterion, which are sensitive to outliers and their performance may be degraded for heavy-tailed noise. In this paper, we go beyond this criterion by investigating the regularized modal regression from a statistical learning viewpoint. A new regularized modal regression model is proposed for estimation and variable selection, which is robust to outliers, heavy-tailed noise, and skewed noise. On the theoretical side, we establish the approximation estimate for learning the conditional mode function, the sparsity analysis for variable selection, and the robustness characterization. On the application side, we applied our model to successfully improve the cognitive impairment prediction using the Alzheimer's Disease Neuroimaging Initiative (ADNI) cohort data.


Properties and applications of calcium hydroxide in endodontics and dental traumatology.
Calcium hydroxide has been included within several materials and antimicrobial formulations that are used in a number of treatment modalities in endodontics. These include, inter-appointment intracanal medicaments, pulp-capping agents and root canal sealers. Calcium hydroxide formulations are also used during treatment of root perforations, root fractures and root resorption and have a role in dental traumatology, for example, following tooth avulsion and luxation injuries. The purpose of this paper is to review the properties and clinical applications of calcium hydroxide in endodontics and dental traumatology including its antibacterial activity, antifungal activity, effect on bacterial biofilms, the synergism between calcium hydroxide and other agents, its effects on the properties of dentine, the diffusion of hydroxyl ions through dentine and its toxicity. Pure calcium hydroxide paste has a high pH (approximately 12.5-12.8) and is classified chemically as a strong base. Its main actions are achieved through the ionic dissociation of Ca(2+) and OH(-) ions and their effect on vital tissues, the induction of hard-tissue deposition and the antibacterial properties. The lethal effects of calcium hydroxide on bacterial cells are probably due to protein denaturation and damage to DNA and cytoplasmic membranes. It has a wide range of antimicrobial activity against common endodontic pathogens but is less effective against Enterococcus faecalis and Candida albicans. Calcium hydroxide is also an effective anti-endotoxin agent. However, its effect on microbial biofilms is controversial.


Rapid growth of a left atrial myxoma shown by echocardiography.
Rapid growth of a left atrial myxoma was demonstrated in a patient who showed no echocardiographic evidence of the tumour when he was admitted for coronary artery bypass grafting. Eight months later he complained of dyspnoea and fatigue, and cross sectional echocardiography showed a 6 x 4 cm left atrial myxoma. This was removed and the diagnosis was confirmed by histological examination.


Clinical practice with anti-dementia drugs: a consensus statement from British Association for Psychopharmacology.
The British Association for Psychopharmacology (BAP) coordinated a meeting of experts to review the evidence on the drug treatment for dementia. The level of evidence (types) was rated using a standard system: Types 1a and 1b (evidence from meta-analysis of randomised controlled trials or at least one controlled trial respectively); types 2a and 2b (one well-designed study or one other type of quasi experimental study respectively); type 3 (non-experimental descriptive studies); and type 4 (expert opinion). There is type 1a evidence for cholinesterase inhibitors (donepezil, rivastigmine and galantamine) for mild to moderate Alzheimer's disease; memantine for moderate to severe Alzheimer's disease; and for the use of bright light therapy and aromatherapy. There is type 1a evidence of no effect of anti inflammatory drugs or statins. There is conflicting evidence regarding oestrogens, with type 2a evidence of a protective effect of oestrogens but 1b evidence of a harmful effect. Type 1a evidence for any effect of B12 and folate will be forthcoming when current trials report. There is type 1b evidence for gingko biloba in producing a modest benefit of cognitive function; cholinesterase inhibitors for the treatment of people with Lewy body disease (particularly neuropsychiatric symptoms); cholinesterase inhibitors and memantine in treatment cognitive impairment associated with vascular dementia; and the effect of metal collating agents (although these should not be prescribed until more data on safety and efficacy are available). There is type 1b evidence to show that neither cholinesterase inhibitors nor vitamin E reduce the risk of developing Alzheimer's disease in people with mild cognitive impairment; and there is no evidence that there is any intervention that can prevent the onset of dementia. There is type 1b evidence for the beneficial effects of adding memantine to cholinesterase inhibitors, and type 2b evidence of positive switching outcomes from one cholinesterase inhibitor to another. There is type 2a evidence for a positive effect of reminiscence therapy, and type 2a evidence that cognitive training does not work. There is type 3 evidence to support the use of psychological interventions in dementia. There is type 2 evidence that a clinical diagnosis of dementia can be made accurately and that brain imaging increases that accuracy. Although the consensus statement dealt largely with medication, the role of dementia care in secondary services (geriatric medicine and old age psychiatry) and primary care, along with health economics, was discussed. There is ample evidence that there are effective treatments for people with dementia, and Alzheimer's disease in particular. Patients, their carers, and clinicians deserve to be optimistic in a field which often attracts therapeutic nihilism.


Single cell analysis of RNA-mediated histone H3.3 recruitment to a cytomegalovirus promoter-regulated transcription site.
Unlike the core histones, which are incorporated into nucleosomes concomitant with DNA replication, histone H3.3 is synthesized throughout the cell cycle and utilized for replication-independent (RI) chromatin assembly. The RI incorporation of H3.3 into nucleosomes is highly conserved and occurs at both euchromatin and heterochromatin. However, neither the mechanism of H3.3 recruitment nor its essential function is well understood. Several different chaperones regulate H3.3 assembly at distinct sites. The H3.3 chaperone, Daxx, and the chromatin-remodeling factor, ATRX, are required for H3.3 incorporation and heterochromatic silencing at telomeres, pericentromeres, and the cytomegalovirus (CMV) promoter. By evaluating H3.3 dynamics at a CMV promoter-regulated transcription site in a genetic background in which RI chromatin assembly is blocked, we have been able to decipher the regulatory events upstream of RI nucleosomal deposition. We find that at the activated transcription site, H3.3 accumulates with sense and antisense RNA, suggesting that it is recruited through an RNA-mediated mechanism. Sense and antisense transcription also increases after H3.3 knockdown, suggesting that the RNA signal is amplified when chromatin assembly is blocked and attenuated by nucleosomal deposition. Additionally, we find that H3.3 is still recruited after Daxx knockdown, supporting a chaperone-independent recruitment mechanism. Sequences in the H3.3 N-terminal tail and αN helix mediate both its recruitment to RNA at the activated transcription site and its interaction with double-stranded RNA in vitro. Interestingly, the H3.3 gain-of-function pediatric glioblastoma mutations, G34R and K27M, differentially affect H3.3 affinity in these assays, suggesting that disruption of an RNA-mediated regulatory event could drive malignant transformation.


The smell of age: perception and discrimination of body odors of different ages.
Our natural body odor goes through several stages of age-dependent changes in chemical composition as we grow older. Similar changes have been reported for several animal species and are thought to facilitate age discrimination of an individual based on body odors, alone. We sought to determine whether humans are able to discriminate between body odor of humans of different ages. Body odors were sampled from three distinct age groups: Young (20-30 years old), Middle-age (45-55), and Old-age (75-95) individuals. Perceptual ratings and age discrimination performance were assessed in 41 young participants. There were significant differences in ratings of both intensity and pleasantness, where body odors from the Old-age group were rated as less intense and less unpleasant than body odors originating from Young and Middle-age donors. Participants were able to discriminate between age categories, with body odor from Old-age donors mediating the effect also after removing variance explained by intensity differences. Similarly, participants were able to correctly assign age labels to body odors originating from Old-age donors but not to body odors originating from other age groups. This experiment suggests that, akin to other animals, humans are able to discriminate age based on body odor alone and that this effect is mediated mainly by body odors emitted by individuals of old age.


Influence of age and sex on left ventricular diastolic strain analysis.
Speckle tracking analysis (STE) of the left ventricle offers a new method to assess left ventricular (LV) diastolic function. LV diastolic strain measurements offer a non-invasive, global and direct measure of LV diastolic function. However, there is little data on normal values and the influence of anthropomorphic factors which is crucial in clinical practice for new techniques. The aims of this study were to formulate reference values for LV diastolic strain rate, elucidate effects of age and sex on LV diastolic strain analysis and compare STE measurements with conventional LV diastolic measurements. One-hundred-forty-seven healthy subjects aged 20-72 years (≥ 28 subjects per age decade) were prospectively included (Mean age 44 ± 13.7 years, 50% female) and examined with electrocardiography and 2D-echocardiography, including speckle tracking. Left ventricular peak early diastolic strain rate (Sre) was measured in the apical windows, using STE. Men had significantly lower LV Sre values than women (1.02 ± 0.22 vs. 1.18 ± 0.23, p value < 0.001). Left ventricular Sre was inversely associated with age, with values decreasing with ageing. An inverse relation was also found with blood pressure and body surface area. Linear regression analysis showed that LV Sre was independently associated with both age and sex. A multivariable linear regression analysis for LV Sre with conventional LV diastolic variables accounted for 70.9% of the variation of LV Sre, showing good model performance. Reference values for LV Sre are reported and found to be both age- and sex-dependent. Therefore we recommend age- and sex-specific references values to be used in daily clinical practice.


Clinical review. Risk factors for low bone mass-related fractures in men: a systematic review and meta-analysis.
Testing men at increased risk for osteoporotic fractures has been recommended. The aim of this study was to estimate the magnitude of association and quality of supporting evidence linking multiple risk factors with low bone mass-related fractures in men. We searched MEDLINE, EMBASE, Web of Science, SCOPUS and Cochrane CENTRAL through February 2010. We identified further studies by reviewing reference lists from selected studies and reviews. Eligible studies had to enroll men and quantitatively evaluate the association of risk factors with low bone density-related fractures. Reviewers working independently and in duplicate determined study eligibility and extracted study description, quality, and outcome data. Fifty-five studies provided data sufficient for meta-analysis. The quality of these observational studies was moderate with fair levels of multivariable adjustment and adequate exposure and outcome ascertainment. Statistically significant associations were established for age, low body mass index, current smoking, excessive alcohol use, chronic corticosteroid use, history of prior fractures, history of falls, history of hypogonadism, history of stroke, and history of diabetes. Statistical heterogeneity of the meta-analytic estimates of all associations was significant except for chronic corticosteroid use. None of these associations were of large magnitude (i.e. adjusted odds ratios were generally <2). No evidence supporting a particular effective testing or screening strategy was identified. Multiple risk factors for fractures in men were identified, but their usefulness for stratifying and selecting men for bone density testing remains uncertain.


Comments on "Factors affecting global flow of scientific knowledge in environmental sciences" by Sonne et al. (2020).
There are major challenges that need to be addressed in the world of scholarly communication, especially in the field of environmental studies and in the context of the United Nations Sustainable Development Goals. Recently, Sonne et al. (2020) published an article in Science of the Total Environment discussing some of these challenges. However, we feel that many of the arguments misrepresent critical elements of Open Access (OA), Plan S, and broader issues in scholarly publishing. In our response, we focus on addressing key elements of their discussion on (i) OA and Plan S, as well as (ii) Open Access Predatory Journals (OAPJ). The authors describe OA and Plan S as restricting author choice, especially through the payment of article-processing charges. The reality is that 'green OA' self-archiving options alleviate virtually all of the risks they mention, and are even the preferred 'routes' to OA as stated by both institutional and national policies in Denmark. In alignment with this, Plan S is also taking a progressive stance on reforming research evaluation. The assumptions these authors make about OA in the "global south" also largely fail to acknowledge some of the progressive work being done in regions like Indonesia and Latin America. Finally, Sonne et al. (2020) highlight the threat that OAPJs face to our scholarly knowledge production system. While we agree generally that OAPJs are problematic, the authors simultaneously fail to mention many of the excellent initiatives helping to combat this threat (e.g., the Directory of Open Access Journals). We call for researchers to more effectively equip themselves with sufficient knowledge of relevant systems before making public statements about them, in order to prevent misinformation from polluting the debate about the future of scholarly communication.


The relationship between chest compression fraction and outcome from ventricular fibrillation arrests in prolonged resuscitations.
Guidelines direct rescuers to minimize CPR interruptions during resuscitation. There is little evidence that evaluates the relationship of increasing CPR fraction among patients with relatively high fractions or prolonged resuscitation. We conducted an observational study of persons who suffered out-of-hospital ventricular fibrillation arrest and required >5 min of emergency medical services (EMS) CPR for persistent pulselessness. We determined the association between hands-on CPR fraction and outcomes of spontaneous circulation, survival to hospital discharge, and neurologically favorable survival. Analyses were stratified by median hands-on CPR and were conducted for those who required 5, 10, and 20 min of EMS CPR for persistent pulselessness. Of 414 potentially eligible patients, 323 (78%) required >5 min of EMS CPR, 234 (56%) required >10 min of EMS CPR, and 153 (37%) required EMS CPR for >20 min. The median CPR fraction was 81%. We did not observe a significant association for the outcomes of hospital survival and neurologically favorable survival for the 5-min and 10-min groups. When restricted to patients who required >20 min of EMS CPR, the half who received a higher hands-on CPR fraction were more likely to achieve spontaneous circulation (40% versus 18%, p=0.004), survival to hospital discharge (20% versus 8%, p=0.03), and neurologically favorable survival (20% versus 7%, p=0.02). Over one-third required 20 min of persistent EMS CPR. The EMS was able to achieve a high hands-on CPR fraction in the context of advanced therapies. Those who required the most prolonged EMS CPR appeared to benefit from greater hands-on CPR fraction.


Two billion years of magmatism recorded from a single Mars meteorite ejection site.
The timing and nature of igneous activity recorded at a single Mars ejection site can be determined from the isotope analyses of Martian meteorites. Northwest Africa (NWA) 7635 has an Sm-Nd crystallization age of 2.403 ± 0.140 billion years, and isotope data indicate that it is derived from an incompatible trace element-depleted mantle source similar to that which produced a geochemically distinct group of 327- to 574-million-year-old "depleted" shergottites. Cosmogenic nuclide data demonstrate that NWA 7635 was ejected from Mars 1.1 million years ago (Ma), as were at least 10 other depleted shergottites. The shared ejection age is consistent with a common ejection site for these meteorites. The spatial association of 327- to 2403-Ma depleted shergottites indicates >2 billion years of magmatism from a long-lived and geochemically distinct volcanic center near the ejection site.


Induction of murine autoimmune disease by chronic polyclonal B cell activation.
In vivo, prolonged polyclonal activation of B cells by the nonantigenic but potent mitogenic lipid A portion of lipopolysaccharide (LPS-R595) resulted in acceleration of the late life systemic lupus erythematosus disease of female MRL/n, BXSB, and NZW mice, mimicking the time, form, and histopathological features characteristic of their early life disease counterparts, i.e., MRL/l females, BXSB males, and (NZB X NZW)F1 females. Similar polyclonal B cell activation of "immunologically normal" mice has less effect and led to a limited expression of autoimmune disease. This R595-induced autoimmunity and immune complex-mediated disease seemed to be the direct result of activation of the immune system and not from other effects of endotoxin since C3H/HeJ, a strain lacking lymphocyte receptors for LPS-R595, had neither serological nor histological evidence of autoimmune disease despite identical treatment.


Clinical relevance of studies on the accuracy of visual inspection for detecting caries lesions: a systematic review.
Although visual inspection is the most commonly used method for caries detection, and consequently the most investigated, studies have not been concerned about the clinical relevance of this procedure. Therefore, we conducted a systematic review in order to perform a critical evaluation considering the clinical relevance and methodological quality of studies on the accuracy of visual inspection for assessing caries lesions. Two independent reviewers searched several databases through July 2013 to identify papers/articles published in English. Other sources were checked to identify unpublished literature. The eligible studies were those which (1) assessed the accuracy of the visual method for detecting caries lesions on occlusal, approximal or smooth surfaces, in primary or permanent teeth, (2) used a reference standard, and (3) reported data about sample size and accuracy of the methods. Aspects related to clinical relevance and the methodological quality of the studies were evaluated. 96 of the 5,578 articles initially identified met the inclusion criteria. In general, most studies failed in considering some clinically relevant aspects: only 1 included study validated activity status of lesions, no study considered its prognosis, 79 studies did not consider a clinically relevant outcome, and only 1 evaluated a patient-centred outcome. Concerning methodological quality, the majority of the studies presented a high risk of bias in sample selection. In conclusion, studies on the accuracy of the visual method for caries detection should consider clinically relevant outcomes besides accuracy; moreover, they should be conducted with higher methodological quality, mainly regarding sample selection.


Carbohydrate-appended photocytotoxic (imidazophenanthroline)-oxovanadium(IV) complexes for cellular targeting and imaging.
Oxovanadium(IV) complexes [VO(aip)(L)](ClO4)2 (L = phtpy, 1; stpy, 2) and [VO(pyip)(L)](ClO4)2 (L = phtpy, 3; stpy, 4), where aip is 2-(9-anthryl)-1H-imidazo[4,5-f][1,10]phenanthroline, pyip is [2-(1-pyrenyl)-1H-imidazo[4,5-f][1,10]phenanthroline, phtpy is (4'-phenyl)-2,2':6',2''-terpyridine and stpy is (2,2':6',2''-terpyridin-4'-oxy)ethyl-β-D-glucopyranoside, were prepared, characterized and their DNA binding and photocleavage activity, cellular uptake and photocytotoxicity in visible light were studied. The complexes are avid binders to calf thymus DNA (K(b) ~10(5) mol(-1)). They efficiently cleave pUC19 DNA in red light of 705 nm via the formation of HO˙ species. The glucose appended complexes 2 and 4 showed higher photocytotoxicity in HeLa and Hep G2 cells over the normal HEK 293T cells. No such preference was observed for the phtpy complexes 1 and 3. No significant difference in IC50 values was observed for the HEK 293T cells. Cell cycle analysis showed that the glucose appended complexes 2 and 4 are more photocytotoxic in cancer cells than in normal cells. Fluorescence microscopy was done to study the cellular localization of complex 4 having a pendant pyrenyl group.


No support for an association with TAAR6 and schizophrenia in a linked population of European ancestry.
Single nucleotide polymorphisms (SNPs) in the trace amine associated receptor trace amine associated receptor 6 gene and 3' flanking region have been shown to be associated with schizophrenia. To replicate these findings, we conducted a family-based association study with the five most significant SNPs in our sample of 79 sib-pair families (56/79 sib-pair families showed linkage to 6q23) and 125 triads. No evidence for association was obtained between these SNPs and schizophrenia in our sample, even when limited to the 56 linked families (P>0.2). We conclude that trace amine associated receptor 6 is not important for the development of schizophrenia in our family samples.


Novel biomarker candidates for gastric cancer.
Gastric cancer continues to be a major threat to human health. Molecular descriptions on the diverse phases of this disease will be valuable for a better diagnosis and development of therapeutic targets. Previously, a 92-gene classifier that distinguishes tumor from non-tumor gastric tissues was proposed. To corroborate this finding, independent approaches of gene selection and class prediction algorithm were applied to the dataset of 86 tissues profiled on 17K cDNA microarrays. As a result, 22 genes were selected, of which 18 were in common with 92 genes previously shown. The differential expression patterns of Chromogranin A (CHGA) and Thy-1 cell surface antigen (THY1) were further validated with immunohistostaining on gastric tissue microarrays. The differential expression patterns of several of the proposed genes have been proven to be critical for tumor progression in other cancer models and will likely function as novel biomarkers for gastric cancer as well.


Transplanted bone marrow mesenchymal stem cells improve memory in rat models of Alzheimer's disease.
The present study aims to evaluate the effect of bone marrow mesenchymal stem cells (MSCs) grafts on cognition deficit in chemically and age-induced Alzheimer's models of rats. In the first experiments aged animals (30 months) were tested in Morris water maze (MWM) and divided into two groups: impaired memory and unimpaired memory. Impaired groups were divided into two groups and cannulated bilaterally at the CA1 of the hippocampus for delivery of mesenchymal stem cells (500 × 10(3)/μL) and PBS (phosphate buffer saline). In the second experiment, Ibotenic acid (Ibo) was injected bilaterally into the nucleus basalis magnocellularis (NBM) of young rats (3 months) and animals were tested in MWM. Then, animals with memory impairment received the following treatments: MSCs (500 × 10(3)/μL) and PBS. Two months after the treatments, cognitive recovery was assessed by MWM in relearning paradigm in both experiments. Results showed that MSCs treatment significantly increased learning ability and memory in both age- and Ibo-induced memory impairment. Adult bone marrow mesenchymal stem cells show promise in treating cognitive decline associated with aging and NBM lesions.


Differing Roles of Functional Movement Variability as Experience Increases in Gymnastics.
Current theories, like Ecological Dynamics, propose that inter-trial movement variability is functional when acquiring or refining movement coordination. Here, we examined how age-based experience levels of gymnasts constrained differences in emergent movement pattern variability during task performance. Specifically, we investigated different roles of movement pattern variability when gymnasts in different age groups performed longswings on a high bar, capturing the range of experience from beginner to advanced status. We also investigated the functionality of the relationships between levels of inter-trial variability and longswing amplitude during performance. One-hundred and thirteen male gymnasts in five age groups were observed performing longswings (with three different experience levels: beginners, intermediates and advanced performers). Performance was evaluated by analysis of key events in coordination of longswing focused on the arm-trunk and trunk-thigh segmental relations. Results revealed that 10 of 18 inter-trial variability measures changed significantly as a function of increasing task experience. Four of ten variability measures conformed to a U-shaped function with age implying exploratory strategies amongst beginners and functional adaptive variability amongst advanced performers. Inter-trial variability of arm-trunk coordination variables (6 of 10) conformed to a \-shaped curve, as values were reduced to complete the longswings. Changes in coordination variability from beginner to intermediate status were largely restrictive, with only one variability measure related to exploration. Data revealed how inter-trial movement variability in gymnastics, relative to performance outcomes, needs careful interpretation, implying different roles as task experience changes. Key pointsInter-trial variability while performing longswings on a high bar was assessed in a large sample (113 participants) divided into five age groups (form beginners to advanced gymnasts). Longswing assessment allowed us to evaluate inter-trial variability in representative performance context.Coordination variability presented two different configurations across experience levels depending on the variable of interest: either a U-shaped or a L- or \-shaped graph.Increased inter-trial variability of the functional phase events offered flexibility to adapt the longswing performance in the advanced gymnasts, while decreasing variability in arm-trunk coordination modes was critical to improve longswing and to achieve the most advanced level.In addition, the relationship between variability measures and the global performance outcome (i.e. the swing amplitude) revealed different functional roles of movement variability (exploratory or restrictive) as a function of changes in experience levels.


Involvement of receptor-interacting protein 2 in innate and adaptive immune responses.
Host defences to microorganisms rely on a coordinated interplay between the innate and adaptive responses of immunity. Infection with intracellular bacteria triggers an immediate innate response requiring macrophages, neutrophils and natural killer cells, whereas subsequent activation of an adaptive response through development of T-helper subtype 1 cells (TH1) proceeds during persistent infection. To understand the physiological role of receptor-interacting protein 2 (Rip2), also known as RICK and CARDIAK, we generated mice with a targeted disruption of the gene coding for Rip2. Here we show that Rip2-deficient mice exhibit a profoundly decreased ability to defend against infection by the intracellular pathogen Listeria monocytogenes. Rip2-deficient macrophages infected with L. monocytogenes or treated with lipopolysaccharide (LPS) have decreased activation of NF-kappaB, whereas dominant negative Rip2 inhibited NF-kappaB activation mediated by Toll-like receptor 4 and Nod1. In vivo, Rip2-deficient mice were resistant to the lethal effects of LPS-induced endotoxic shock. Furthermore, Rip2 deficiency results in impaired interferon-gamma production in both TH1 and natural killer cells, attributed in part to defective interleukin-12-induced Stat4 activation. Our data reflect requirements for Rip2 in multiple pathways regulating immune and inflammatory responses.


Is sperm freezability related to the post-thaw lipid peroxidation and the formation of reactive oxygen species in boars?
The aim of the present study was to determine whether the levels of reactive oxygen species (ROS) substances production and the levels of lipid peroxidation of the sperm membrane were related to the quality that the ejaculates exhibited after cryopreservation in boars. Ejaculates from 42 healthy boars were used in this study and they were cryopreserved with the lactose-egg yolk extender (LEY). Several sperm quality parameters were assessed by flow cytometry in samples incubated for 30 and 150 min at 37 °C after thawing: the percentage of sperm with intact plasma membrane (SIPM), intracellular reactive oxygen substances production through mean of DCF fluorescence intensity of total sperm (mean-DCF) and the percentage of viable and non-viable sperm containing oxidized BODIPY (VSOB and NVSOB). In addition, the percentages of total motile (TMS) and progressively motile sperm (PMS) were assessed at the same incubation times with a computer-assisted sperm analysis system. The classification of the ejaculates into good or bad freezers was performed through hierarchical cluster analysis from SIPM and TMS at 150 min post-thawing. The ejaculates of those males classified as good freezers exhibited higher (p < 0.05) SPIM, TMS and PMS than the bad freezers, although both groups presented similar (p > 0.05) VSOB, NVSOB and mean-DCF. Therefore, these results show that lipid peroxidation and the amount of reactive oxygen substances in the sperm after cryopreservation are similar between boars classified as good or bad freezers.


Correlates of objectively assessed physical activity and sedentary time in children: a cross-sectional study (The European Youth Heart Study).
Identifying leisure time activities performed before and after school that influence time in physical activity (PA) and/or time spent sedentary can provide useful information when designing interventions aimed to promote an active lifestyle in young people. The purpose of this study was to examine associations between mode of transportation to school, outdoor play after school, participation in exercise in clubs, and TV viewing with objectively assessed PA and sedentary behaviour in children. A total of 1327 nine- and 15-year-old children from three European countries (Norway, Estonia, Portugal) participated as part of the European Youth Heart Study. PA was measured during two weekdays and two weekend days using the MTI accelerometer, and average percent of time in moderate-to-vigorous PA (MVPA) and time spent sedentary were derived. Potential correlates were assessed by self-report. Independent associations between self-reported correlates with percent time in MVPA and percent time sedentary were analysed by general linear models, adjusted by age, gender, country, measurement period, monitored days and parental socio-economic status. In 9-year-olds, playing outdoors after school was associated with higher percent time in MVPA (P < 0.01), while participation in sport clubs was associated with higher percent time in MVPA (P < 0.01) in 15-year-olds. No associations with percent time sedentary were observed in either age group. Frequency of outdoor play after school is a significant correlate for daily time in MVPA in 9-year-olds, while this correlate is attenuated in favour of participation in sport and exercise in clubs in 15-year-olds. Targeting walking to school or reduced TV viewing time in order to increase time in daily MVPA in children is unlikely to be sufficient. Correlates related to time spent sedentary need further examination.


Inhibition of AP-1 and MAPK signaling and activation of Nrf2/ARE pathway by quercitrin.
Quercitrin, glycosylated form of flavonoid compounds, is widely distributed in nature. Extensive studies have demonstrated that quercitrin exhibits strong antioxidant and anti-carcinogenic activities. However, the molecular mechanism is poorly understood. The present study examines the effects of quercitrin on tumor promotion in mouse JB6 cells, a validated model for screening cancer chemopreventive agents and elucidating the molecular mechanisms. Quercitrin blocked TPA-induced neoplastic transformation in JB6 P+ cells. Pretreatment of JB6 cells with quercitrin down-regulated transactivation of AP-1 and NF-kappaB induced by UVB or TPA. In the skin of AP-1-luciferase transgenic mice, topical treatment of the mouse with quercitrin markedly blocked the TPA-induced AP-1 transactivation. Further studies indicated that these inhibitory actions appear to be mediated through the inhibition of MAPKs phosphorylation, including ERKs, p38 kinase, and JNKs. In addition, quercitrin stimulated the activation of NF-E2-related factor (Nrf2) and GST ARE-luciferase activity. Comet assays showed that quercitrin could block DNA damage induced by UVB. To our knowledge, these results provide the first evidence that quercitrin contributes to the inhibition of neoplastic transformation by blocking activation of the MAPK pathway and stimulation of cellular protection signaling. Moreover, to our knowledge, these findings provide the first molecular basis for the anti-carcinogenic action of quercitrin.


CAIX and pax-8 Commonly Immunoreactive in Endolymphatic Sac Tumors: A Clinicopathologic Study of 26 Cases with Differential Considerations for Metastatic Renal Cell Carcinoma in von Hippel-Lindau Patients.
Endolymphatic sac tumors (ELSTs) are rare, slowly growing temporal bone neoplasms which show a high association with von Hippel-Lindau (VHL) syndrome. The immunohistochemistry evaluation of these papillary-cystic neoplasms frequently raises the differential diagnosis with renal cell carcinoma, among other metastatic neoplasms, whether in VHL patients or not. A cohort of 26 patients with ELSTs were evaluated for histologic features, immunohistochemistry findings, and association with VHL. Standard immunohistochemistry evaluation was performed. Sixteen females and 10 males ranging in age from 10 to 69 years (mean 44; VHL mean: 32) at initial presentation, comprised the cohort of patients. Most (86%) experienced hearing changes or inner ear symptoms (vertigo, dizziness), with an average duration of symptoms for 39 months (range 2-240 months). The tumors were an average of 2.9 cm (range 0.4-8 cm), with 14 left, 11 right sided and one bilateral tumor. Nine patients had documented VHL, with 3 patients having a concurrent or subsequent clear cell renal cell carcinoma. Patients were followed an average of 6.2 years (available in 24 patients): 19 alive without disease, 7.5 years; 2 dead without disease, 1.2 years; and 3 alive with disease, 3.1 years. The neoplastic cells show the following immunohistochemistry findings: AE1/AE3, EMA, CK7, CAIX, GLUT1, VEGF: 100% of cases tested were positive; pax-8: 85% of cases positive; CD10 and RCC: 0% of cases reactive. Based on this cohort of 26 patients with ELST, 9 of whom had VHL, the strong pax-8 and CAIX should be used in conjunction with negative CD10 and RCC to help exclude a metastatic renal cell carcinoma. As CAIX is an enzyme overexpressed in hypoxia and hypoxia inducible factor is what VHL protein regulates, this is an expected, although previously unreported finding. Whether part of VHL or not, VHL mutations may be a somatic rather than germline finding in the tumors, a possible further explanation for the CAIX reaction.


Is lower hip range of motion a risk factor for groin pain in athletes? A systematic review with clinical applications.
Whether hip range of motion (ROM) is a risk factor for groin pain in athletes is not known. To systematically review the relationship between hip ROM and groin pain in athletes in cross-sectional/case-control and prospective studies. Systematic review, prospectively registered (PROSPERO) according to PRISMA guidelines. Pubmed, Embase, CINAHL and SPORTDiscus were systematically searched up to December 2015. Two authors performed study selection, data extraction/analysis, quality assessment (Critical Appraisal Skills Programme) and strength of evidence synthesis. We identified seven prospective and four case-control studies. The total quality score ranged from 29% to 92%. Heterogeneity in groin pain classification, injury definitions and physical assessment precluded data pooling. There was strong evidence that total rotation of both hips below 85° measured at the pre-season screening was a risk factor for groin pain development. Strong evidence suggested that internal rotation, abduction and extension were not associated with the risk or presence of groin pain. Total hip ROM is the factor most consistently related to groin pain in athletes. Screening for hip ROM is unlikely to correctly identify an athlete at risk of developing groin pain because of the small ROM differences found and poor ROM measurement properties.


CD20 deficiency in humans results in impaired T cell-independent antibody responses.
CD20 was the first B cell differentiation antigen identified, and CD20-specific mAbs are commonly used for the treatment of B cell malignancies and autoantibody-mediated autoimmune diseases. Despite this the role of CD20 in human B cell physiology has remained elusive. We describe here a juvenile patient with CD20 deficiency due to a homozygous mutation in a splice junction of the CD20 gene (also known as MS4A1) that results in "cryptic" splicing and nonfunctional mRNA species. Analysis of this patient has led us to conclude that CD20 has a central role in the generation of T cell-independent (TI) antibody responses. Key evidence to support this conclusion was provided by the observation that although antigen-independent B cells developed normally in the absence of CD20 expression, antibody formation, particularly after vaccination with TI antigens, was strongly impaired in the patient. Consistent with this, TI antipolysaccharide B cell responses were severely impeded in CD20-deficient mice. Our study therefore identifies what we believe to be a novel type of humoral immunodeficiency caused by CD20 deficiency and characterized by normal development of antigen-independent B cells, along with a reduced capacity to mount proper antibody responses.


Seasonal to decadal variability in ice discharge from the Greenland Ice Sheet.
Rapid changes in thickness and velocity have been observed at many marine-terminating glaciers in Greenland, impacting the volume of ice they export, or discharge, from the ice sheet. While annual estimates of ice-sheet wide discharge have been previously derived, higher-resolution records are required to fully constrain the temporal response of these glaciers to various climatic and mechanical drivers that vary in sub-annual scales. Here we sample outlet glaciers wider than 1 km (N = 230) to derive the first continuous, ice-sheet wide record of total ice sheet discharge for the 2000-2016 period, resolving a seasonal variability of 6 %. The amplitude of seasonality varies spatially across the ice sheet from 5 % in the southeastern region to 9 % in the northwest region. We analyze seasonal to annual variability in the discharge time series with respect to both modelled meltwater runoff, obtained from RACMO2.3p2, and glacier front position changes over the same period. We find that year-to-year changes in total ice sheet discharge are related to annual front changes (r 2 = 0.59, p = 10-4) and that the annual magnitude of discharge is closely related to cumulative front position changes (r 2 = 0.79), which show a net retreat of > 400 km, or an average retreat of > 2 km at each surveyed glacier. Neither maximum seasonal runoff or annual runoff totals are correlated to annual discharge, which suggests that larger annual quantities of runoff do not relate to increased annual discharge. Discharge and runoff, however, follow similar patterns of seasonal variability with near-coincident periods of acceleration and seasonal maxima. These results suggest that changes in glacier front position drive secular trends in discharge, whereas the impact of runoff is likely limited to the summer months when observed seasonal variations are substantially controlled by the timing of meltwater input.


Geminin prevents DNA damage in vagal neural crest cells to ensure normal enteric neurogenesis.
In vertebrate organisms, the neural crest (NC) gives rise to multipotential and highly migratory progenitors which are distributed throughout the embryo and generate, among other structures, the peripheral nervous system, including the intrinsic neuroglial networks of the gut, i.e. the enteric nervous system (ENS). The majority of enteric neurons and glia originate from vagal NC-derived progenitors which invade the foregut mesenchyme and migrate rostro-caudally to colonise the entire length of the gut. Although the migratory behaviour of NC cells has been studied extensively, it remains unclear how their properties and response to microenvironment change as they navigate through complex cellular terrains to reach their target embryonic sites. Using conditional gene inactivation in mice we demonstrate here that the cell cycle-dependent protein Geminin (Gem) is critical for the survival of ENS progenitors in a stage-dependent manner. Gem deletion in early ENS progenitors (prior to foregut invasion) resulted in cell-autonomous activation of DNA damage response and p53-dependent apoptosis, leading to severe intestinal aganglionosis. In contrast, ablation of Gem shortly after ENS progenitors had invaded the embryonic gut did not result in discernible survival or migratory deficits. In contrast to other developmental systems, we obtained no evidence for a role of Gem in commitment or differentiation of ENS lineages. The stage-dependent resistance of ENS progenitors to mutation-induced genotoxic stress was further supported by the enhanced survival of post gut invasion ENS lineages to γ-irradiation relative to their predecessors. Our experiments demonstrate that, in mammals, NC-derived ENS lineages are sensitive to genotoxic stress in a stage-specific manner. Following gut invasion, ENS progenitors are distinctly resistant to Gem ablation and irradiation in comparison to their pre-enteric counterparts. These studies suggest that the microenvironment of the embryonic gut protects ENS progenitors and their progeny from genotoxic stress.


Caffeine as a lead compound for the design of therapeutic agents for the treatment of Parkinson's disease.
The current pharmacological therapies for the treatment of Parkinson's disease are mostly inadequate and recent, improved therapeutic agents are required. Two important molecular targets for the design of anti-parkinsonian therapeutic compounds are the adenosine A2A receptor and the enzyme, monoamine oxidase (MAO) B. Adenosine A2A receptor antagonists are a relatively new class of anti-parkinsonian agents, which act by potentiating dopamine-mediated neurotransmission via dopamine D2 receptors. MAO-B inhibitors are established therapy of Parkinson's disease and inhibit the MAO-B-catalysed metabolism of dopamine in the brain. This conserves reduced dopamine stores and extends the action of dopamine. A2A antagonism and MAO-B inhibition have also been associated with neuroprotective effects, further establishing roles for these classes of compounds in Parkinson's disease. Interestingly, caffeine, a known adenosine receptor antagonist, has been recently considered as a lead compound for the design and discovery of A2A antagonists and MAO-B inhibitors. This review summarizes the recent efforts to discover caffeinederived MAO-B inhibitors. The design of caffeine-derived A2A antagonists has been extensively reviewed previously. The prospect of discovering dual-target-directed compounds that act at both targets is also evaluated. Compounds that block the activation and function of both A2A receptors and MAO-B may have a synergistic effect in the treatment of patients with Parkinson's disease.


Effects of black cohosh and estrogen on core body and tail-skin temperatures in ovariectomized rats by telemetric monitoring with dual thermistor probes.
To investigate the effects of black cohosh and estrogen on the temperature in ovariectomized rats, the core body temperature (CBT) and tail-skin temperature (TST) were simultaneously monitored and the relationship between these two temperatures was explored. Twenty-four female Sprague-Dawley rats aged 8 weeks were randomly divided into four groups: sham-operated (SHAM), ovariectomized (OVX), OVX treated with estradiol valerate (OVX + E), and OVX treated with isopropanolic black cohosh extract (OVX + ICR). Rats were sham-operated or ovariectomized and were implanted with telemetry transmitters with dual thermistor probes. Two weeks after surgery, the animals were treated with drugs for 4 weeks. During the last week of the treatments, the dynamic temperature profiles of the CBT and TST were collected. The average CBT and TST, TST fluctuation frequency, and the average amplitude fluctuation were significantly higher in OVX than in SHAM rats. In addition, dramatic fluctuations of TST in OVX rats occurred at the time points of the day when the CBTs were lower in OVX rats than in SHAM rats. Treatment of OVX rats with estradiol valerate or isopropanolic black cohosh extract markedly decreased the average CBT and TST, TST fluctuation frequency, and the average amplitude fluctuation. Moreover, CBT was found to be significantly higher, while TST was lower in OVX + E than in OVX + ICR rats. Both black cohosh and estradiol treatments ameliorated the abnormal thermoregulation in OVX rats. In particular, black cohosh reduced CBT better than estradiol and estradiol reduced TST better than black cohosh.


International expert workshop on the analysis of the economic and public health impacts of air pollution: workshop summary.
Forty-nine experts from 18 industrial and developing countries met on 6 September 2001 in Garmisch-Partenkirchen, Germany, to discuss the economic and public health impacts of air pollution, particularly with respect to assessing the public health benefits from technologies and policies that reduce greenhouse gas (GHG) emissions. Such measures would provide immediate public health benefits, such as reduced premature mortality and chronic morbidity, through improved local air quality. These mitigation strategies also allow long-term goals--for example, reducing the buildup of GHG emissions--to be achieved alongside short-term aims, such as immediate improvements in air quality, and therefore benefits to public health. The workshop aimed to foster research partnerships by improving collaboration and communication among various agencies and researchers; providing a forum for presentations by sponsoring agencies and researchers regarding research efforts and agency activities; identifying key issues, knowledge gaps, methodological shortcomings, and research needs; and recommending activities and initiatives for research, collaboration, and communication. This workshop summary briefly describes presentations made by workshop participants and the conclusions of three separate working groups: economics, benefits transfer, and policy; indoor air quality issues and susceptible populations; and development and transfer of dose-response relationships and exposure models in developing countries. Several common themes emerged from the working group sessions and subsequent discussion. Key recommendations include the need for improved communication and extended collaboration, guidance and support for researchers, advances in methods, and resource support for data collection, assessment, and research.


[The auditory perception of frequency-localized acoustic on-events].
The quick changing of amplitude envelope (amplitude modulation) is the inherent properties of speech signals. They are analyzed by the auditory system and are reflected in the auditory sensation as a sequences of some "events", that participate in creating of phonetic images. The dynamics of frequency distribution of events is considered to be used by listeners as a determinants of some consonants. However, this supposition are inconsistent with the psychoacoustics experiments showed that listeners can't determine the frequency of amplitude irregularities of tonal components. Current work consists of the discussion of this problem and results of experiments where the perception of an amplitude jumps in different frequency format area was studied using the synthetic quasi--syllable. The results obtained give evidence that the phoneme interpretations "know" the frequency of amplitude irregularieties.


Culture shock and healthcare workers in remote Indigenous communities of Australia: what do we know and how can we measure it?
Culture shock or cultural adaptation is a significant issue confronting non-Indigenous health professionals working in remote Indigenous communities in Australia. This article is presented in two parts. The first part provides a thorough background in the theory of culture shock and cultural adaptation, and a comprehensive analysis of the consequences, causes, and current issues around the phenomenon in the remote Australian healthcare context. Second, the article presents the results of a comprehensive literature review undertaken to determine if existing studies provide tools which may measure the cultural adaptation of remote health professionals. A comprehensive literature review was conducted utilising the meta-databases CINAHL and Ovid Medline. While there is a plethora of descriptive literature about culture shock and cultural adaptation, empirical evidence is lacking. In particular, no empirical evidence was found relating to the cultural adaptation of non-Indigenous health professionals working in Indigenous communities in Australia. In all, 15 international articles were found that provided empirical evidence to support the concept of culture shock. Of these, only 2 articles contained tools that met the pre-determined selection criteria to measure the stages of culture shock. The 2 instruments identified were the Culture Shock Profile (CSP) by Zapf and the Culture Shock Adaptation Inventory (CSAI) by Juffer. There is sufficient evidence to determine that culture shock is a significant issue for non-Indigenous health professionals working in Indigenous communities in Australia. However, further research in this area is needed. The available empirical evidence indicates that a measurement tool is possible but needs further development to be suitable for use in remote Indigenous communities in Australia.


Evaluating an Internet Gaming Disorder Scale Using Mokken Scaling Analysis.
Internet Gaming Disorder (IGD) was recently included as a condition for further study in the fifth and latest version of the Diagnostic and Statistical Manual of Mental Disorders. The present study investigated whether the IGD criteria comprise a unidimensional construct. Data stemmed from a sample of Norwegians aged 17.5 years in 2012 and 19.5 years in 2014 (N = 1258). The study used the Mokken scale analysis to investigate whether the score of the different items on the IGD scale measured a single latent variable and if the scale functions differently for males and females. Correlation analysis was conducted between the scores on the IGD scale (count) and the Gaming Addiction Scale for Adolescents (GASA, categorical), both assessed in 2014. Negative binomial regression analyses were applied in order to investigate how different predictors of mental health assessed in 2012 were associated with IGD assessed in 2014. The Mokken scale analysis showed that all item-coefficients of homogeneity exceeded 0.3 when the whole sample completed the scale and when females completed the scale, indicating that the items reflect a single latent variable. In both cases moderate (H > 0.40) unidimensionality was shown. The item measuring "tolerance" did not exceed 0.3 in the scale when completed by males, indicating that only eight out of nine items reflect a single latent variable when applied to males only. The eight-item scale containing males showed weak (H > 0.30) unidimensionality. The correlation analysis showed a positive correlation between the scores on the IGD scale and the GASA (r = 0.71, p < 0.01) when assessed simultaneously and a positive but lower correlation (r = 0.48, p < 0.01) when assessed longitudinally. Results from the negative binomial regression analysis showed that previous video-game addiction, being male, depression, aggression and loneliness were significant predictors of IGD. The associations were small for all independent variables except previous video game addiction and gender where the associations were large. Although the results from the correlation analysis and regression analysis showed predictive validity of the scale, the results from the Mokken analysis suggest that the IGD scale may not be applied as a unidimensional scale when the tolerance item is included.


Changes in Serial D-Dimer Levels Predict the Prognoses of Trousseau's Syndrome Patients.
Background: The development of acute multiple embolic infarctions (AMEI) resulting from cancer is known as Trousseau's syndrome (TS). At present, however, there is no good marker for predicting the prognosis of TS patients. In the present study, we evaluated the use of serial D-dimer levels as a prognostic marker for TS. Methods: This retrospective cohort study included 1,409 consecutive acute ischemic stroke patients. We selected a group of patients with TS showing AMEI (n = 38; TS group) and a group of patients with atrial fibrillation (Af) and AMEI (n = 35; Af group) as controls. Serial D-dimer levels were measured between days 7 and 28 after stroke (sub-acute phase) in 21 patients of the TS group and 24 patients of the Af group. Results: D-dimer levels at onset (acute phase) were significantly higher in the TS group (8.45 ± 1.79 μg/mL, n = 38) compared with the Af group (1.14 ± 0.14 μg/mL, n = 35) (p < 0.0001). In patients for whom serial D-dimer measurements were made, D-dimer levels measured at the sub-acute phase decreased to 0.48 ± 0.12 μg/mL (n = 24) in the Af group, but remained elevated in the TS group during the sub-acute phase (11.20 ± 2.77 μg/mL, n = 21) (p < 0.0001). In all TS patients in whom serial D-dimer measurements were made, D-dimer levels in 17 patients who died within 500 days (13.31 ± 3.23 μg/mL) were significantly higher than those of the four surviving patients (2.23 ± 0.38 μg/mL) (cut-off D-dimer level = 3.0 μg/mL) during this period. Moreover, serial D-dimer levels of 10 patients who died within 90 days (17.78 ± 4.60 μg/mL) were significantly higher than those of the 11 patients who survived up to 90 days (5.21 ± 2.12 μg/mL) (p < 0.05). Conclusions: Serial D-dimer levels may be a good biomarker for TS as well as a useful predictor of the prognosis of TS patients.


The use of conventional and complementary health services and self-prescribed treatments amongst young women with constipation: An Australian national cohort study.
Little research has been conducted regarding the comprehensive health service utilisation in constipation care. This study investigates the comprehensive health service utilisation amongst Australian women with constipation. This study draws upon data from the Australian Longitudinal Study on Women's Health. A total of 8074 young women were asked about their frequency of constipation, measures of quality of life, and use of a range of health services and self-prescribed treatments via two postal surveys conducted in 2006 and 2009, respectively. The prevalence of constipation was 18.5% amongst women in 2009. Constipated women had poorer quality of health than women without constipation. Women who sought help for constipation were more likely to visit multiple groups of conventional and complementary health practitioners compared to women who did not experience constipation (p<0.005). However, women were less likely to visit a specialist for the management of constipation over time (2006 to 2009). There was an increase in the proportion of women with constipation who self-prescribed vitamins/minerals over time (p<0.001). Although only 4.5% of women sought help for their constipation, given the increasing use of multiple health services across time, more studies are required regarding the optimal treatment in constipation care.


Helping each other to learn--a process evaluation of peer assisted learning.
The benefits of Peer Assisted Learning (PAL) are well established with positive effects on examination scores, student satisfaction and personal and professional development reported. PAL is increasingly utilised as a resource within medical education where the restrictions on resources have forced teachers to look at creating new educational environments which can be delivered at a lower cost. This study sought to evaluate the processes at work as the emphasis of PAL research to date has largely been on the consideration of student outcomes. Fifth-year medical undergraduates, who had completed their communication skills modular training and attended a preparatory workshop, facilitated a role-play session for their second-year colleagues within an Early Patient Contact programme. Semi-structured interviews and focus groups were used to collect data at different time points in order to establish the views of peer learners and tutors towards this new method of teaching. The data was analysed according to the principles framework analysis using N-vivo software. Themes were shared and debated with the multidisciplinary team of authors and a concordance of views on common themes was reached after discussion and debate. Analysis of the data resulted in the emergence of three thematic categories: Learning Environment, Educational Exchange and Communication and Modelling. The data demonstrated a concordance of the views between peer tutors and learners on barriers and levers of this approach as well as a heightened awareness of the learning environment and the educational exchange occurring therein. The data is significant as it not only demonstrates a high level of acceptability among tutors and learners for PAL but also indicates the reciprocity of educational exchange that appears to occur within the PAL setting. This study highlights some of the unique characteristics of PAL and we recommend the development of further qualitative studies around peer learners and tutors views of this process.


Gastroenteritis in children: principles of diagnosis and treatment.
Gastroenteritis in children is a common reason for visits to family physicians. Most cases of gastroenteritis have a viral etiology and are self-limited. However, more severe or prolonged cases of gastroenteritis can result in dehydration with significant morbidity and mortality. This is often the scenario in third-world countries, where gastroenteritis results in 3 million deaths annually. A proper clinical evaluation will allow the physician to estimate the percentage of dehydration and determine appropriate therapy. In some situations, laboratory studies such as determination of blood urea nitrogen and serum electrolytes may be helpful. Stool studies are indicated if a child is having bloody diarrhea or if an unusual etiology is suspected, such as Escherichia coli O157:H7 or Cryptosporidium. Most children with gastroenteritis can be treated with physiologically balanced oral rehydration solutions. In children who are hypovolemic, lethargic and estimated to be more than 5 percent dehydrated, initial treatment with intravenous boluses of isotonic saline or Ringer's lactate may be required. Children with severe diarrhea need nutrition to restore digestive function and, generally, food should not be withheld.


The Role of Culex pipiens L. (Diptera: Culicidae) in Virus Transmission in Europe.
Over the past three decades, a range of mosquito-borne viruses that threaten public and veterinary health have emerged or re-emerged in Europe. Mosquito surveillance activities have highlighted the Culex pipiens species complex as being critical for the maintenance of a number of these viruses. This species complex contains morphologically similar forms that exhibit variation in phenotypes that can influence the probability of virus transmission. Critical amongst these is the choice of host on which to feed, with different forms showing different feeding preferences. This influences the ability of the mosquito to vector viruses and facilitate transmission of viruses to humans and domestic animals. Biases towards blood-feeding on avian or mammalian hosts have been demonstrated for different Cx. pipiens ecoforms and emerging evidence of hybrid populations across Europe adds another level of complexity to virus transmission. A range of molecular methods based on DNA have been developed to enable discrimination between morphologically indistinguishable forms, although this remains an active area of research. This review provides a comprehensive overview of developments in the understanding of the ecology, behaviour and genetics of Cx. pipiens in Europe, and how this influences arbovirus transmission.


Yleaf: Software for Human Y-Chromosomal Haplogroup Inference from Next-Generation Sequencing Data.
Next-generation sequencing (NGS) technologies offer immense possibilities given the large genomic data they simultaneously deliver. The human Y-chromosome serves as good example how NGS benefits various applications in evolution, anthropology, genealogy, and forensics. Prior to NGS, the Y-chromosome phylogenetic tree consisted of a few hundred branches, based on NGS data, it now contains many thousands. The complexity of both, Y tree and NGS data provide challenges for haplogroup assignment. For effective analysis and interpretation of Y-chromosome NGS data, we present Yleaf, a publically available, automated, user-friendly software for high-resolution Y-chromosome haplogroup inference independently of library and sequencing methods.


ATP regulation of type-1 inositol 1,4,5-trisphosphate receptor activity does not require walker A-type ATP-binding motifs.
ATP is known to increase the activity of the type-1 inositol 1,4,5-trisphosphate receptor (InsP3R1). This effect is attributed to the binding of ATP to glycine rich Walker A-type motifs present in the regulatory domain of the receptor. Only two such motifs are present in neuronal S2+ splice variant of InsP3R1 and are designated the ATPA and ATPB sites. The ATPA site is unique to InsP3R1, and the ATPB site is conserved among all three InsP3R isoforms. Despite the fact that both the ATPA and ATPB sites are known to bind ATP, the relative contribution of these two sites to the enhancing effects of ATP on InsP3R1 function is not known. We report here a mutational analysis of the ATPA and ATPB sites and conclude neither of these sites is required for ATP modulation of InsP3R1. ATP augmented InsP3-induced Ca2+ release from permeabilized cells expressing wild type and ATP-binding site-deficient InsP3R1. Similarly, ATP increased the single channel open probability of the mutated InsP3R1 to the same extent as wild type. ATP likely exerts its effects on InsP3R1 channel function via a novel and as yet unidentified mechanism.


Changes in Frailty Status and Risk of Depression: Results From the Progetto Veneto Anziani Longitudinal Study.
To evaluate whether prefrailty was associated with the risk of developing depression and if longitudinal changes in frailty status corresponded to changes in incident depression during follow up. A population-based, prospective cohort study was conducted for 4.4 years in two separate geographic areas near the city of Padua in the Veneto Region of Northern Italy. In 891 nondepressed, nonfrail, community-dwelling Italian subjects aged ≥ 65 (46.6% men) belonging to the Progetto Veneto Anziani study, depression was defined according to the Geriatric Depression Scale and was confirmed by geriatricians skilled in psychogeriatric medicine. Prefrailty was defined by the presence of one or two criteria among the Fried criteria. The incidence rate of depression was 13.3% among subjects improving their frailty status at follow-up (N = 15), 15.0% in those who remained stable (N = 79), and 26.7% among worsening participants (N = 67) (p = 0.001). Prefrailty at baseline did not predict the onset of depression (HR: 0.82; 95% CI: 0.55-1.21; Wald χ2 = 0.73; df = 1; p = 0.43), but a deterioration during follow-up in at least one additional frailty criteria was associated with a significantly higher risk (HR: 1.95; 95% CI: 1.32-2.89; Wald χ2 = 5.78; df = 2; p = 0.01). Improvement in frailty status was not associated with the risk of incident depression (HR: 0.71; 95% CI: 0.35-1.42; Wald χ2 = 0.47; df = 2; p = 0.28). Our data did not offer evidence that prefrailty per se predisposes to the onset of depression, but worsening in frailty status is associated with an almost twofold increased risk of incident depression, irrespective from the initial level of impairment.


Human Goodpasture anti-alpha3(IV)NC1 autoantibodies share structural determinants.
To examine the structural relationship among autoantibodies produced by individuals with anti-GBM antibody-mediated disease, a polyclonal anti-idiotype directed against human anti-alpha3(IV)NC1 antibodies was produced and then used to study autoantibodies from other patients. For this purpose, anti-alpha3(IV)NC1 antibodies (anti-GBM), derived from a single patient (LL) with high titer and typical anti-GBM antibody specificity, were isolated using recombinant alpha3(IV)NC1-sepharose affinity chromatography. Following hyperimmunization of rabbits with anti-GBM IgG, irrelevant rabbit anti-human IgG antibodies were removed from the antiserum using a human IgG-sepharose column. The rabbit anti-alpha3(IV)NC1 antibodies (anti-Id GBM) effluent bound to human anti-GBM antibodies, but it did not bind to either normal human IgG or recombinant alpha3(IV)NC1 protein. The Id-anti-Id interaction was conformationally dependent on intact heavy and light chains of the anti-alpha3(IV)NC1 antibodies (ELISA and Western blotting). A competitive immunoassay was developed to evaluate structural and potential genetic relationships among anti-alpha3(IV)NC1 antibodies from different patients. All patients tested (9 of 9) had a substantial fraction (producing > 50% inhibition) of anti-GBM antibodies expressing Id-GBM. The results indicate that shared determinants are expressed by anti-GBM antibodies from different individuals, and they raise the possibility that common genetic elements are used to encode them. These regions are potential targets for design of reagents to regulate autoreactive B cells and/or interfere with pathogenic antibody-GBM interactions, in individuals with anti-GBM antibody mediated diseases.


The teaching of clinical skills in the context of children's nursing: a UK survey.
The teaching of clinical skills within nursing is currently enjoying a resurgence following the implementation of the 'Fitness for Practice Curriculum'. However, the teaching of clinical skills specifically within children's nursing has received little attention within the research arena. This article presents the data of a national postal survey sent to United Kingdom institutions providing pre-registration child branch studies, to ascertain the current provision of clinical skills teaching. The findings conclude that very few institutions have invested in specific skills laboratories to teach children's nursing students. It was also identified that a wide range of clinical skills are taught to child branch students; however, the resources and realism to clinical practice is limited by the lack of specialist children's skills laboratories or equipment. In conclusion the authors recommend the development of child specific skills laboratories, to augment child branch skills teaching, in order to enhance the realistic simulation of clinical practice.


Immediate cortical adaptation in visual and non-visual areas functions induced by monovision.
Monovision is an optical correction for presbyopes that consists of correcting one eye for far distance and the other for near distance, creating a superimposition of an in-focus with a blurred image. Brain adaptation to monovision was studied in unexperienced observers by measuring visual evoked potentials from 64-channels. The first clear effect of monovision on visual evoked potentials was the C1 amplitude reduction, indicating that the unilateral blurring induced by monovision reduces feed-forward activity in primary visual area. Monovision led also to an increased amplitude of the P1 and pP1 components, with the latter originating in prefrontal regions. This effect probably works as an attentional compensatory activity used to compensate for the degraded V1 signal. A common and often successful option to correct presbyopia with contact lenses is monovision. This is an unbalanced correction across the two eyes where one eye is corrected for far vision and the other eye is corrected for near vision. Monovision is therefore a form of acquired anisometropia that causes a superimposition of an in-focus image with a blurred image. In spite of this visual anisometropia, monovision has been successfully used for many decadesl however the brain mechanism supporting monovision is not well understood. The present study aimed to measure the visual evoked potentials with a high-density electrode array (64-channel) in a group of presbyopes and to provide a detailed spatiotemporal analysis of the cortical activity after a short period of adaptation to monovision with contact lenses. When compared with a balanced eye near correction, monovision produced both a clear reduction of the earliest visual evoked potential components, the C1 and the N1, and an amplitude increase of the P1 and pP1. These results indicate that the unilateral blurring induced by wearing monovision contact lenses reduces feed-forward activity in the primary visual area and feedback activity in extrastriate areas (C1 and N1 reduction). Interestingly, other brain activities in both extrastriate visual areas (the P1 component) and in the anterior insula (the pP1 component) appear to compensate for this dysfunction, increasing their activity during monovision. These changes confirm the presence of fluid brain adaptation in visual and non-visual areas during monocular interferences.


Antiglycation and antioxidation properties of Juglans regia and Calendula officinalis: possible role in reducing diabetic complications and slowing down ageing.
Accumulation of advanced glycation end products (AGEs) in the body due to the non-enzymatic glycation of proteins and oxidation is associated with aging and diabetes mellitus. In this study we wanted to investigate the antiglycation and antioxidation potential of two medicinal plants: Juglans regia and Calendula officinalis. In-vitro investigation was carried out to discover the antiglycation and antioxidation potential of J. regia and C. officinalis. Using an Ultraviolet Double-beam Spectrophotometer, we evaluated the antiglycation property of the crude methanolic extracts of J. regia and C. officinalis by assessing their ability to inhibit the Maillard reaction. Employing the same instrument we also measured the antioxidation potential of these plant extracts using the nitric oxide (NO) free radical-scavenging assay. J. regia had greater antiglycation ability, with a minimum inhibitory concentration (MIC50) of 28 microg/mL as compared with that of C. officinalis (270 microg/mL). C. officinalis had greater antioxidation potential (26.10, 22.07 and 16.06% at 0.5 mg, 0.25 mg and 0.125 mg, respectively, as compared with 18.15, 16.50 and 16.06% of J. regia, respectively). J. regia and C. officinalis inhibited the Maillard reaction and prevented oxidation in-vitro. Hence, the extracts of these plants could have therapeutic uses in curbing chronic diabetic complications and slowing down aging.


Platelet kinetics in the pulmonary microcirculation in vivo assessed by intravital microscopy.
Growing evidence supports the substantial pathophysiological impact of platelets on the development of acute lung injury. Methods for studying these cellular mechanisms in vivo are not present yet. The aim of this study was to develop a model enabling the quantitative analysis of platelet kinetics and platelet-endothelium interaction within consecutive segments of the pulmonary microcirculation in vivo. New Zealand White rabbits were anesthetized and ventilated. Autologous platelets were separated from blood and labeled ex vivo with rhodamine 6G. After implantation of a thoracic window, microhemodynamics and kinetics of platelets were investigated by intravital microscopy. Velocities of red blood cells (RBCs) and platelets were measured in arterioles, capillaries and venules, and the number of platelets adhering to the microvascular endothelium was counted. Kinetics of unstimulated platelets was compared with kinetics of thrombin-activated platelets. Velocity of unstimulated platelets was comparable to RBC velocity in all vessel segments. Unstimulated platelets passed the pulmonary microcirculation without substantial platelet-endothelial interaction. In contrast, velocity of activated platelets was decreased in all vascular segments indicating platelet margination and temporal platelet-endothelium interaction. Thrombin-activated platelets adhered to arteriolar endothelium; in capillaries and venules adherence of platelets was increased 8-fold and 13-fold, respectively. In conclusion, using intravital microscopy platelet kinetics were directly analyzed in the pulmonary microcirculation in vivo for the first time. In contrast to leukocytes, no substantial platelet-endothelium interaction occurs in the pulmonary microcirculation without any further stimulus. In response to platelet activation, molecular mechanisms enable adhesion of platelets in arterioles and venules as well as retention of platelets within capillaries.


mRNA-specific reverse transcription-polymerase chain reaction from human tissue extracts.
Reverse transcription-polymerase chain reaction (RT-PCR) has become the method of choice for detection of mRNA transcripts, including those of low abundance obtained from small precious samples of human tissue. A major confounding problem for standard reverse-transcription-priming strategies is the presence of contaminating genomic DNA (gDNA) carried over from the original "RNA" extract into the RT and PCR steps. The contaminating gDNA contains a processed pseudogene sequence-which lacks introns but contains a poly(A) tail-for commonly studied internal reference genes beta-actin and GAPDH, and target genes GSTM1, GSTP1, and others. These pseudogene sequences therefore confound standard-design "RNA-specific" PCR primer pairs which rely, for cDNA versus gDNA specificity, on the pair-spanning introns, or one of the individual primer oligos spanning an exon/exon splice site, because these features are lacking in processed pseudogene sequences. The result is false RT-PCR positives for these "housekeeper" genes in total RNA extracts; the gDNA processed pseudogene is mistaken for mRNA gene transcript. A universal RT primer has been designed that targets the poly(A) tail of mRNA and adds a unique tag sequence not otherwise existing in the human genome. Genomic DNA does not incorporate this RT-inserted unique tag. PCR is then performed using a transcript-specific forward primer and a reverse primer that is identical to the unique tag incorporated at RT. Only cDNA made with this RT primer is compatible with this reverse PCR primer, thus eliminating confounding signal from contaminating gDNA. This method performs RNA-specific qualitative and quantitative evaluation of gene expression, while preserving the sensitivity of standard RT-PCR techniques. Applications to low-copy transcripts in human samples are demonstrated.


A review of residential radon case-control epidemiologic studies performed in the United States.
Lung cancer is the leading cause of cancer death in the United States for both men and women. Although most lung cancer deaths are attributable to tobacco usage, even secondary causes of lung cancer are important because of the magnitude of lung cancer incidence and its poor survival rate. This review summarizes the basic features and major findings from the published U.S. large-scale residential radon case-control studies performed in New Jersey, Iowa, and Missouri (two studies). The methodology from an unpublished study covering Connecticut, Utah, and Southern Idaho is also presented. Overall, the higher categorical risk estimates for these published studies produced a positive association between prolonged radon exposure and lung cancer. Two studies (Missouri-II and Iowa) that incorporated enhanced dose estimates produced the most compelling evidence suggesting an association between prolonged residential radon exposure and lung cancer. The prevailing evidence suggests that the statistically significant findings may be related to improved retrospective radon exposure estimates. The general findings from the U.S. studies, along with extrapolations from radon-exposed underground miners, support the conclusion that after cigarette smoking, prolonged residential radon exposure is the second leading cause of lung cancer in the general population.


Hepatitis C virus core protein interacts with the cytoplasmic tail of lymphotoxin-beta receptor.
Hepatitis C virus (HCV) core protein is a multifunctional protein. We examined whether it can interact with cellular proteins, thus contributing to viral pathogenesis. Using the HCV core protein as a bait to screen a human liver cDNA library in a yeast two-hybrid screening system, we have isolated several positive clones encoding cellular proteins that interact with the HCV core protein. Interestingly, more than half of these clones encode the cytoplasmic domain of lymphotoxin-beta receptor (LT betaR), which is a member of the tumor necrosis factor receptor family. Their binding was confirmed by in vitro glutathione S-transferase fusion protein binding assay and protein-protein blotting assay to be direct and specific. The binding sites were mapped within a 58-amino-acid region of the cytoplasmic tail of LT betaR. The binding site in the HCV core protein was localized within amino acid residues 36 to 91 from the N terminus, corresponding to the hydrophilic region of the protein. In mammalian cells, the core protein was found to be associated with the membrane-bound LT betaR. Since the LT betaR is involved in germinal center formation and developmental regulation of peripheral lymphoid organs, lymph node development, and apoptotic signaling, the binding of HCV core protein to LT betaR suggests the possibility that this viral protein has an immunomodulating function and may explain the mechanism of viral persistence and pathogenesis of HCV.