The impact of fragility fracture and approaches to osteoporosis risk assessment worldwide.
Osteoporosis constitutes a major public health problem, through its association with age-related fractures, particularly of the hip, vertebrae, distal forearm and humerus. Substantial geographic variation has been noted in the incidence of osteoporotic fractures worldwide, with Western populations (North America, Europe and Oceania), reporting increases in hip fracture throughout the second half of the 20th century, with a stabilisation or decline in the last two decades. In developing populations however, particularly in Asia, the rates of osteoporotic fracture appears to be increasing. The massive global burden consequent to osteoporosis means that fracture risk assessment should be a high priority among health measures considered by policy makers. The WHO operational definition of osteoporosis, based on a measurement of bone mineral density (BMD) by dual-energy X-ray absorptiometry (DXA), has been used globally since the mid-1990s. However, although this definition identifies those at greatest individual risk of fracture, in the population overall a greater total number of fractures occur in individuals with BMD values above the threshold for osteoporosis diagnosis. A number of web-based tools to enable the inclusion of clinical risk factors, with or without BMD, in fracture prediction algorithms have been developed to improve the identification of individuals at high fracture risk, the most commonly used globally being FRAX®. Access to DXA, osteoporosis risk assessment, case finding and treatment varies worldwide, but despite such advances studies indicate that a minority of men and women at high fracture risk receive treatment. Importantly, research is ongoing to demonstrate the clinical efficacy and cost-effectiveness of osteoporosis case finding and risk assessment strategies worldwide. The huge burden caused by osteoporosis related fractures to individuals, healthcare systems and societies should provide a clear impetus for the progression of such approaches.


Potential proteolytic activity of fibronectin: fibronectin laminase and its substrate specificity.
The purified 190-kDa fibronectin fragment produced by cathepsin D can be spontaneously activated in the presence of CaCl2. This activation generates new proteolytic activities and also results in the formation of several subfragments. One of them exhibits the activity of FN-gelatinase that preferentially splits type I denatured collagen and fibronectin (see preceding paper). In this work we describe the purification and characterization of another fragment (25 kDa), issued from the same autodigest. This fragment may be activated to yield another proteinase, that splits preferentially laminin and denatured collagen type I. This enzyme will be referred as FN-laminase. Purified FN-laminase specifically reacted with antibodies against fibronectin. The specificity of bond cleavage by FN-laminase was studied with various synthetic peptides analogous to collagen repeats. FN-laminase cleaves the Ala-Gly bond in the sequence GPAGPR; the arginine residue in position P3' is important for this cleavage. The enzyme is inhibited by pepstatin A and phenylmethanesulfonyl fluoride, like retroviral aspartic proteinases. It is also inhibited by EDTA. No inhibition was obtained with 1,10-phenanthroline or 4-chloromercuribenzoate, inhibitors of Zn-metalloproteinases or cysteine proteinases, respectively.


Measuring the mechanical properties of molecular conformers.
Scanning probe-actuated single molecule manipulation has proven to be an exceptionally powerful tool for the systematic atomic-scale interrogation of molecular adsorbates. To date, however, the extent to which molecular conformation affects the force required to push or pull a single molecule has not been explored. Here we probe the mechanochemical response of two tetra(4-bromophenyl)porphyrin conformers using non-contact atomic force microscopy where we find a large difference between the lateral forces required for manipulation. Remarkably, despite sharing very similar adsorption characteristics, variations in the potential energy surface are capable of prohibiting probe-induced positioning of one conformer, while simultaneously permitting manipulation of the alternative conformational form. Our results are interpreted in the context of dispersion-corrected density functional theory calculations which reveal significant differences in the diffusion barriers for each conformer. These results demonstrate that conformational variation significantly modifies the mechanical response of even simple porpyhrins, potentially affecting many other flexible molecules.


Long-term exposure to electric fields. A cross-sectional epidemiologic investigation of occupationally exposed workers in high-voltage substations.
In the present epidemiologic study, 53 workers with a long-term (more than five years) exposure to the electric field of 400 kV substations were examined and compared with a matched reference group of 53 nonexposed workers from the same power companies. Matching variables included age, geographic location and employment time. The aim of the study was to investigate the possibility of persistent, chronic health effects in the exposed group as a consequence of exposure. The investigation indluded the nervous system (neurasthenic symptoms, psychological tests, electroencephalography), the cardiovascular system (symptoms, blood pressure, electrocardiography), and the blood (hemoglobin, red blood cells, reticulocytes, white blood cells including differential count, thrombocytes, sedimentation rate). Fertility was also assessed. The results showed no differences between the exposed and reference groups as a consequence of the long-term exposure to the electric fields. The groups differed, however, in that the exposed group had (a) consistently better results on the psychological performance tests, (b) a fewer number of children, especially boys, and (c) somewhat higher education. The differences in test results were due to the higher education among the exposed. The difference in number of children was also thought to be related to factors other than exposure since it was found to be present already 10--15 years before the work in 400 kV substations began.


Acute respiratory distress syndrome: A rare presentation of amantadine toxicity.
Male, 64 FINAL DIAGNOSIS: Acute Respiratory Distress Syndrome (ARDS) Symptoms: Generalized myoclonic jerks • impaired concentration • memory decline • visual hallucinations Amantadine HCl Clinical Procedure: - Specialty: Toxicology. Adverse events of drug therapy. Amantadine is indicated for treatment of influenza A infection, Parkinson disease and extrapyramidal reactions. Amantadine overdose affects mainly cardiovascular and central nervous systems. Amantadine-induced respiratory failure has not been described in previous case reports but it is a potential known side effect. We describe the case of a 64-year-old African American male with end stage renal disease who was prescribed amantadine at a dose for normal kidney function (300 milligrams per day) for no clear reasons. PATIENT's serum level of amantadine drawn on admission was found to be 6200 nanogram per deciliter (ng/dl) with normal range being 700-1000 ng/dl. Amantadine hydrochloride is not actively metabolized in humans; is mainly excreted unchanged in urine by glomerular filtration and tubular secretion (90% of the ingested dose). It tends to accumulate in patients with impaired renal function; poorly excreted in patients on hemodialysis and has a large volume of distribution. Our patient with impaired renal function was prescribed a much higher dose and eventually presented with high serum concentration of amantadine and neurological manifestations suggestive of amantadine toxicity. He developed sudden onset respiratory failure and pulmonary edema which is described as a potential lethal complication of amantadine toxicity. Since there is no specific etiology for his respiratory failure, this could represent the first reported case of Amantadine-induced Adult Respiratory Distress Syndrome (ARDS).


Demographic and clinical profile of substance abusing women seeking treatment at a de-addiction center in north India.
In the recent decades increasing number of women have been seeking deaddiction services. Despite that the report data is very limited from India. The present research aimed to study the demographic and clinical profile of women seeking deaddiction treatment at a tertiary care center in North India. Retrospective structured chart review of 100 women substance abusers seeking treatment at a deaddiction center between September 1978 and December 2011. A typical case was of 36.3 years age, married (65%), urban (61%), nuclear family (59%) based housewife (56%), with good to fair social support (69%). The commonest substance of abuse was tobacco (60%), followed by opioids (27%), alcohol (15%), and benzodiazepines (13%). The common reasons for initiation of substance use were to alleviate frustration or stress (49%) and curiosity (37%). Family history of drug dependence (43%), comorbidity (25%), and impairments in health (74%), family (57%), and social domains (56%) were common. Only a third of the sample paid one or more follow visit, and of those 58% were abstinent at the last follow-up. Significant predictors identified were being non-Hindu and higher educational years for abstinent status at follow-up. The common substances of abuse were tobacco, opioids, and alcohol and benzodiazepines; and family history of drug abuse and comorbidity were common. The follow-up and outcome were generally poor. This profile gives us some clues to address a hidden health problem of the community.


Stabilizing Morbidity and Predicting the Aesthetic Results of Radial Forearm Free Flap Donor Sites.
The radial forearm flap is a versatile, widely used flap. However, the possibility of donor site complications has led to concern over its use. Some surgeons prefer using other flaps whose donor sites can be closed primarily with less morbidity, including avoiding unpleasant scarring. However, in our experience, donor site stability of the radial forearm flap can be reliably achieved by using well-implemented specific procedures. Here, we present a collection of donor site cases of the radial forearm flap and investigate factors that affect the aesthetic results as the basis for a reference for selecting a radial forearm flap. In this retrospective study, we reviewed 171 cases in which a radial forearm flap was used for free tissue transfer after resecting head and neck cancer. We focused on donor site morbidity rates. Each operation involved a detailed procedure designed to minimize donor site morbidity. Moreover, statistical investigations were conducted for 22 cases to determine factors affecting the scar appearance. Only one case developed total skin graft necrosis as a major complication. Scar-related aesthetic results were acceptable, and the body-mass index, body weight, diabetes, and cardiac problems were significant factors related to the appearance of scars. Performing the radial forearm flap using a well-implemented detailed technique helps achieve acceptable donor site morbidity results. The aesthetic results were more promising for patients without excess body weight, diabetes, or cardiac problems. Therefore, anxiety about donor site morbidity should not be a reason to avoid selecting the radial forearm flap in suitable patients.


Mechanical circulatory support for advanced heart failure: effect of patient selection on outcome.
Use of wearable left ventricular assist systems (LVAS) in the treatment of advanced heart failure has steadily increased since 1993, when these devices became generally available in Europe. The aim of this study was to identify in an unselected cohort of LVAS recipients those aspects of patient selection that have an impact on postimplant survival. Data were obtained from the Novacor European Registry. Between 1993 and 1999, 464 patients were implanted with the Novacor LVAS. The majority had idiopathic (60%) or ischemic (27%) cardiomyopathy; the median age at implant was 49 (16 to 75) years. The median support time was 100 days (4.1 years maximum). Forty-nine percent of the recipients were discharged from the hospital on LVAS; they spent 75% of their time out of the hospital. For a subset of 366 recipients, for whom a complete set of data was available, multivariate analysis revealed that the following preimplant conditions were independent risk factors for survival after LVAS implantation: respiratory failure associated with septicemia (odds ratio 11.2), right heart failure (odds ratio 3.2), age >65 years (odds ratio 3.01), acute postcardiotomy (odds ratio 1.8), and acute infarction (odds ratio 1.7). For patients without any of these factors, the 1-year survival after LVAS implantation including the posttransplantation period was 60%; for the combined group with at least 1 risk factor, it was 24%. Careful selection, specifically implantation before patients become moribund, and improvement of management may result in improved outcomes of LVAS treatment for advanced heart failure.


Towards Horizon 2020: challenges and advances for clinical mental health research - outcome of an expert survey.
The size and increasing burden of disease due to mental disorders in Europe poses substantial challenges to its population and to the health policy of the European Union. This warrants a specific research agenda concerning clinical mental health research as one of the cornerstones of sustainable mental health research and health policy in Europe. The aim of this research was to identify the top priorities needed to address the main challenges in clinical research for mental disorders. The research was conducted as an expert survey and expert panel discussion during a scientific workshop. Eighty-nine experts in clinical research and representing most European countries participated in this survey. Identified top priorities were the need for new intervention studies, understanding the diagnostic and therapeutic implications of mechanisms of disease, and research in the field of somatic-psychiatric comorbidity. The "subjectivity gap" between basic neuroscience research and clinical reality for patients with mental disorders is considered the main challenge in psychiatric research, suggesting that a shift in research paradigms is required. Innovations in clinical mental health research should bridge the gap between mechanisms underlying novel therapeutic interventions and the patient experience of mental disorder and, if present, somatic comorbidity. Clinical mental health research is relatively underfunded and should receive specific attention in Horizon 2020 funding programs.


Occupational exposure to radio frequency/microwave radiation and the risk of brain tumors: Interphone Study Group, Germany.
It is still under debate whether occupational exposure to radio frequency/microwave electromagnetic fields (RF/MW-EMF) contributes to the development of brain tumors. This analysis examined the role of occupational RF/MW-EMF exposure in the risk of glioma and meningioma. A population-based, case-control study including 381 meningioma cases, 366 glioma cases, and 1,494 controls aged 30-69 years was performed in three German regions in 2000-2003. An exposure matrix for occupational activity was constructed by using information on RF/MW-EMF exposure collected in a computer-assisted personal interview. "High" exposure was defined as an occupational exposure that may exceed the RF/MW-EMF exposure limits for the general public recommended by the International Commission on Non-Ionizing Radiation Protection. Multiple conditional logistic regressions were performed separately for glioma and meningioma. No significant association between occupational exposure to RF/MW-EMF and brain tumors was found. For glioma, the adjusted odds ratio for highly exposed persons compared with persons not highly exposed was 1.21 (95% confidence interval: 0.69, 2.13); for meningioma, it was 1.34 (95% confidence interval: 0.64, 2.81). However, the slight increase in risk observed with increasing duration of exposure merits further research with larger sample sizes.


Enhanced Neural Responses to Imagined Primary Rewards Predict Reduced Monetary Temporal Discounting.
The pervasive tendency to discount the value of future rewards varies considerably across individuals and has important implications for health and well-being. Here, we used fMRI with human participants to examine whether an individual's neural representation of an imagined primary reward predicts the degree to which the value of delayed monetary payments is discounted. Because future rewards can never be experienced at the time of choice, imagining or simulating the benefits of a future reward may play a critical role in decisions between alternatives with either immediate or delayed benefits. We found that enhanced ventromedial prefrontal cortex response during imagined primary reward receipt was correlated with reduced discounting in a separate monetary intertemporal choice task. Furthermore, activity in enhanced ventromedial prefrontal cortex during reward imagination predicted temporal discounting behavior both between- and within-individual decision makers with 62% and 73% mean balanced accuracy, respectively. These results suggest that the quality of reward imagination may impact the degree to which future outcomes are discounted. Significance statement: We report a novel test of the hypothesis that an important factor influencing the discount rate for future rewards is the quality with which they are imagined or estimated in the present. Previous work has shown that temporal discounting is linked to individual characteristics ranging from general intelligence to the propensity for addiction. We demonstrate that individual differences in a neurobiological measure of primary reward imagination are significantly correlated with discounting rates for future monetary payments. Moreover, our neurobiological measure of imagination can be used to accurately predict choice behavior both between and within individuals. These results suggest that improving reward imagination may be a useful therapeutic target for individuals whose high discount rates promote detrimental behaviors.


Unraveling the anxious mind: anxiety, worry, and frontal engagement in sustained attention versus off-task processing.
Much remains unknown regarding the relationship between anxiety, worry, sustained attention, and frontal function. Here, we addressed this using a sustained attention task adapted for functional magnetic resonance imaging. Participants responded to presentation of simple stimuli, withholding responses to an infrequent "No Go" stimulus. Dorsolateral prefrontal cortex (DLPFC) activity to "Go" trials, and dorsal anterior cingulate (dACC) activity to "No Go" trials were associated with faster error-free performance; consistent with DLPFC and dACC facilitating proactive and reactive control, respectively. Trait anxiety was linked to reduced recruitment of these regions, slower error-free performance, and decreased frontal-thalamo-striatal connectivity. This indicates an association between trait anxiety and impoverished frontal control of attention, even when external distractors are absent. In task blocks where commission errors were made, greater DLPFC-precuneus and DLPFC-posterior cingulate connectivity were associated with both trait anxiety and worry, indicative of increased off-task thought. Notably, unlike trait anxiety, worry was not linked to reduced frontal-striatal-thalamo connectivity, impoverished frontal recruitment, or slowed responding during blocks without commission errors, contrary to accounts proposing a direct causal link between worry and impoverished attentional control. This leads us to propose a new model of the relationship between anxiety, worry and frontal engagement in attentional control versus off-task thought.


Ecosystem nitrogen fixation throughout the snow-free period in subarctic tundra: effects of willow and birch litter addition and warming.
Nitrogen (N) fixation in moss-associated cyanobacteria is one of the main sources of available N for N-limited ecosystems such as subarctic tundra. Yet, N2 fixation in mosses is strongly influenced by soil moisture and temperature. Thus, temporal scaling up of low-frequency in situ measurements to several weeks, months or even the entire growing season without taking into account changes in abiotic conditions cannot capture the variation in moss-associated N2 fixation. We therefore aimed to estimate moss-associated N2 fixation throughout the snow-free period in subarctic tundra in field experiments simulating climate change: willow (Salix myrsinifolia) and birch (Betula pubescens spp. tortuosa) litter addition, and warming. To achieve this, we established relationships between measured in situ N2 fixation rates and soil moisture and soil temperature and used high-resolution measurements of soil moisture and soil temperature (hourly from May to October) to model N2 fixation. The modelled N2 fixation rates were highest in the warmed (2.8 ± 0.3 kg N ha-1 ) and birch litter addition plots (2.8 ± 0.2 kg N ha-1 ), and lowest in the plots receiving willow litter (1.6 ± 0.2 kg N ha-1 ). The control plots had intermediate rates (2.2 ± 0.2 kg N ha-1 ). Further, N2 fixation was highest during the summer in the warmed plots, but was lowest in the litter addition plots during the same period. The temperature and moisture dependence of N2 fixation was different between the climate change treatments, indicating a shift in the N2 fixer community. Our findings, using a combined empirical and modelling approach, suggest that a longer snow-free period and increased temperatures in a future climate will likely lead to higher N2 fixation rates in mosses. Yet, the consequences of increased litter fall on moss-associated N2 fixation due to shrub expansion in the Arctic will depend on the shrub species' litter traits.


Establishing insulin 1 and insulin 2 knockout congenic strains on NOD genetic background.
As insulin is a major autoantigen in autoimmune diabetes and because the insulin gene region locus in humans has been linked to diabetes risk, we have bred insulin gene knockouts onto the NOD mouse. Mice differ from humans in terms that they express two nonallelic genes of insulin. Insulin 2 is the murine homologue of the human insulin gene and is located on mouse chromosome 7. Insulin 1 is thought to have evolved by a gene duplication event, lacks the second intron of the insulin 2 gene, and is located on mouse chromosome 19. The differential thymic expression of the insulin gene may be important for central tolerance induction. Here, we present the initial establishment of congenic knockouts and characterization of the congenic intervals corresponding to insulin 1 and insulin 2 knockout genes on mouse chromosome 19 and 7, respectively.


Mechanisms of Argon Retention in Clays Revealed by Laser 40Ar-39Ar Dating.
A method for dating clays is important for studies of weathering, diagenesis, hydrocarbon migration, and the formation of major metalliferous deposits. However, many attempts have produced imprecise or inaccurate results. Data from shales show that, contrary to expectations, the (40)Ar-(39)Ar dating technique can be successfully used to determine the diagenetic age of ancient sediments because (39)Ar losses during irradiation are controlled by release from low retentivity sites in illite equivalent to those that have lost radiogenic (40)Ar in nature, rather than by direct recoil as is generally assumed.


Signals and structural features involved in integral membrane protein targeting to the inner nuclear membrane.
We have examined transfected cells by immunofluorescence microscopy to determine the signals and structural features required for the targeting of integral membrane proteins to the inner nuclear membrane. Lamin B receptor (LBR) is a resident protein of the nuclear envelope inner membrane that has a nucleoplasmic, amino-terminal domain and a carboxyl-terminal domain with eight putative transmembrane segments. The amino-terminal domain of LBR can target both a cytosolic protein to the nucleus and a type II integral protein to the inner nuclear membrane. Neither a nuclear localization signal (NLS) of a soluble protein, nor full-length histone H1, can target an integral protein to the inner nuclear membrane although they can target cytosolic proteins to the nucleus. The addition of an NLS to a protein normally located in the inner nuclear membrane, however, does not inhibit its targeting. When the amino-terminal domain of LBR is increased in size from approximately 22.5 to approximately 70 kD, the chimeric protein cannot reach the inner nuclear membrane. The carboxyl-terminal domain of LBR, separated from the amino-terminal domain, also concentrates in the inner nuclear membrane, demonstrating two nonoverlapping targeting signals in this protein. Signals and structural features required for the inner nuclear membrane targeting of proteins are distinct from those involved in targeting soluble polypeptides to the nucleoplasm. The structure of the nucleocytoplasmic domain of an inner nuclear membrane protein also influences targeting, possibly because of size constraints dictated by the lateral channels of the nuclear pore complexes.


Blebbistatin protects iPSC-CMs from hypercontraction and facilitates automated patch-clamp based electrophysiological study.
Recently, there have been great advances in cardiovascular channelopathy modeling and drug safety pharmacology using human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs). The automated patch-clamp (APC) technique overcomes the disadvantages of the manual patch-clamp (MPC) technique, which is labor intensive and gives low output. However, the application of the APC platform is still limited in iPSC-CM based research, due to the difficulty in maintaining the high quality of single iPSC-CMs during dissociation and recording. In this study, we improved the method for single iPSC-CM preparation by applying 2.5 µM blebbistatin (BB, an excitation-contraction coupling uncoupler) throughout APC procedures (dissociation, filtration, storage, and recording). Under non-BB buffered condition, iPSC-CMs in suspension showed a severe bleb-like morphology. However, BB-supplement led to significant improvements in morphology and INa recording, and we even obtained several CMs that showed spontaneous action potentials with typical morphology. Furthermore, APC faithfully recapitulated the single-cell electrophysiological phenotypes of iPSC-CMs derived from Brugada syndrome patients, as detected with MPC. Our study indicates that APC is capable of replacing MPC in the modeling of cardiac channelopathies using human iPSC-CMs by providing high-quality data with higher throughput.


Covalently bonded multimers of Au25(SBut)18 as a conjugated system.
Aromatic dithiol linkers were used to prepare aggregates of Au25(SR)18 clusters (SR: thiolate) via ligand exchange reactions. Fractions of different aggregate sizes were separated by size exclusion chromatography (SEC). The aggregates were characterized by UV-vis absorption spectroscopy, matrix assisted laser desorption ionization (MALDI) mass spectrometry, nuclear magnetic resonance (NMR) spectroscopy (including diffusion-ordered spectroscopy, DOSY) and small angle X-ray scattering (SAXS). At a 2 : 1 cluster : dithiol ratio, small aggregates (dimers, trimers) and also larger aggregates consisting of 10-20 Au25 clusters were formed, according to DOSY, besides unreacted (monomeric) Au25(SR)18. MALDI mass spectrometry shows signals consistent with dimers and trimers (doubly charged). The SAXS curves for the small aggregates can be well fitted by a pearl-necklace model. For the bigger aggregates the SAXS curves evidence a characteristic separation distance between the clusters within the aggregates, which is imposed by the length of the linker. The SAXS curves of these larger aggregates can be well fitted with a core-shell sphere model with a sticky hard-sphere structure factor, in agreement with closely packed aggregates. The absorption spectra of smaller aggregates resemble those of individual Au25(SR)18 clusters; however, and most importantly, the larger aggregates show completely different, less structured spectra with a new band emerging at 840 nm. We assign this drastic change in the absorption spectra and the new band to the electronic coupling between the clusters through the all aromatic linker. In accordance with this view, the aggregates formed with a linker containing methylene groups, thus breaking conjugation, do not show the band at 840 nm. By the addition of monothiols to the larger aggregates their size can be reduced through an "unlinking" reaction. This reaction also affects the band at 840 nm, which moves to higher energy when reducing the aggregate size, as would be expected within a particle in a box model. The electronic coupling between the clusters through the linker is the basis for future applications in nanoelectronics.


Design and self-assembly of simple coat proteins for artificial viruses.
Viruses are among the simplest biological systems and are highly effective vehicles for the delivery of genetic material into susceptible host cells. Artificial viruses can be used as model systems for providing insights into natural viruses and can be considered a testing ground for developing artificial life. Moreover, they are used in biomedical and biotechnological applications, such as targeted delivery of nucleic acids for gene therapy and as scaffolds in material science. In a natural setting, survival of viruses requires that a significant fraction of the replicated genomes be completely protected by coat proteins. Complete protection of the genome is ensured by a highly cooperative supramolecular process between the coat proteins and the nucleic acids, which is based on reversible, weak and allosteric interactions only. However, incorporating this type of supramolecular cooperativity into artificial viruses remains challenging. Here, we report a rational design for a self-assembling minimal viral coat protein based on simple polypeptide domains. Our coat protein features precise control over the cooperativity of its self-assembly with single DNA molecules to finally form rod-shaped virus-like particles. We confirm the validity of our design principles by showing that the kinetics of self-assembly of our virus-like particles follows a previous model developed for tobacco mosaic virus. We show that our virus-like particles protect DNA against enzymatic degradation and transfect cells with considerable efficiency, making them promising delivery vehicles.


Long-term followup comparing two treatment dosing strategies of (125) I plaque radiotherapy in the management of small/medium posterior uveal melanoma.
Objective. To investigate the efficacy of two different dosing strategies of radioactive iodine-125 ((125)I) in the management of small- and medium-sized posterior uveal melanoma. Patients and Methods. The medical records of consecutive patients with choroidal melanomas between 1.5 and 5.0 mm in apical height treated initially with (125)I plaque radiotherapy were reviewed. Patients were treated with one of the following two treatment dosing strategies: (1) 85 Gy to the apical height of the tumor (group 1) or (2) 85 Gy to a prescription point of 5.0 mm (group 2). Results. Of 95 patients, 55 patients were treated to the apical height of the tumor, and 40 were treated to a prescription point of 5.0 mm. Comparative analysis of the incidence rates of specific complications between the two groups demonstrates that group 2 had a significantly higher incidence of radiation retinopathy, radiation optic neuropathy, and/or visually significant cataract formation than group 1 (P = 0.028). Conclusion. Treatment of choroidal melanomas less than 5 mm in apical height with (125)I brachytherapy to the true apical height is equally effective when compared to treatment with 85 Gy to 5.0 mm. Treatment to the apical height of the tumor may result in lower incidence of radiation-related complications.


A profile of substance abusers using the emergency services in a tertiary care hospital in Sikkim.
Sikkim, a state in Northeast India with a population of more than 500,000 and inhabited by indigenous population of Lepchas, Bhutias and Nepalis, lies in the foothills of the Himalayas sharing borders with Tibet, Nepal and Bhutan. Northeast India is a major source of injection drug users (IDUs) and associated HIV/AIDS. Alcohol use is traditionally prevalent in Sikkim and recently, IDU behaviour has also been reported, although systematic information on epidemiology and treatment availability of substance abuse in Sikkim is not available. To study the sociodemographic and drug use profile of substance abusers using the emergency services in a tertiary care hospital. A retrospective chart review was used. Patients with history of current drug use seeking emergency services for any medical or surgical consequence incident to substance abuse from July 2000 to June 2005 (60 months) were included in the study. Data were generated from emergency case register, hospital records and case sheets. SPSS 10.0 was used for data analysis. Out of 54 patients seeking emergency services with substance abuse (1.16% of all psychiatric consultations), alcohol abusers were 77.8% and other opioid abusers 14.8%. Prevalence of IDU was 16.66%. Common opioids abused were dextrpropoxyphene and pentazocine, both analgesics. A significant number of patients (46.3%) had a history of >20 days/month frequency of abuse. Median of duration of abuse with all drugs was 12 years, while that with IDU population was 3 years. Alcohol withdrawal was the commonest cause (57.4%) of reporting to the emergency. Psychiatric comorbidity was found among 7.4%. Commonest medications used were chlordiazepoxide and clonidine, for withdrawal and naltrexone, for substitution. No standardized treatment protocol for substitution treatment was available. This is an initial attempt to study the sociodemographic and drug use profile of substance abusers in Sikkim. Demographic and socioeconomic characteristics of substance abusers seeking emergency services are not significantly different from treatment-seeking substance abusers in other parts of India. IDU behaviour has been detected and low median duration of use suggests an emerging problem and need for urgent harm reduction. Alcohol withdrawal was the commonest cause of seeking emergency services, which is related to high prevalence of alcohol abuse in Sikkim. No standardized substitution treatment is available for substance abusers, which may lead to higher rates of relapse.


An alternative histochemical method to simultaneously demonstrate muscle nuclei and muscle fibre type.
We present a modified histochemical method to examine, simultaneously, nuclei and fibre type in human skeletal muscle. The new procedure (Haem-ATPase) is based on two previously used histochemical protocols. Biopsies were obtained from the rectus abdominis muscle of patients undergoing elective abdominal surgery. Fibre type composition, cross-sectional area (CSA) and nuclei to fibre ratio (N:F) were determined from frozen sections of each biopsy. To test the validity of the new method, serial sections of each biopsy were stained separately using the standard and modified methods. No differences were found in fibre type distribution, mean-weighted CSA and N:F when comparing the modified method with the standard methods. The Haem-ATPase method was found to shrink fibre size by at least 3% ( P>0.05) compared with the established myosin acid labile method. We propose that this modified technique is suitable for initial examination of both the nuclei and fibre type in the same frozen sections of human skeletal muscle.


Pediatric laparoscopic surgery; initial experience from Pakistan; first 100 cases in single center.
In a developing country like Pakistan, laparoscopic surgeries are not considered favourable by many, possibly because of high costs or a lack of expertise. It is an established fact that laparoscopic surgery offers better surgical treatments with a shorter hospital stay and fewer complications. The current retrospective study was conducted at a tertiary care hospital in Karachi and comprised of laparoscopy cases performed by a single surgeon from March 2012 to September of 2014. A total of 100laparoscopic surgeries were performed; mostly appendectomies 49(49%) and undescended testes (UDTs) 34(34%). Overall, there were 70(70%) male patients. The mean age of the patients was 7.1 years and standard deviation (SD) of 2. Four (4%) patients had cellulitis. Laparoscopy paediatric surgery offered advantages of fewer wound-associated complications, less incisional pain, a shorter recovery time, and improved cosmesis.


Whole-body CT in polytrauma patients: effect of arm positioning on thoracic and abdominal image quality.
The purpose of this study is to assess the influence of different arm positioning techniques on thoracic and abdominal image quality and radiation dose of whole-body trauma CT (wbCT). One hundred and fifty polytrauma patients (104 male, mean age 47 ± 19) underwent wbCT with arms elevated above the head (group A, n=50), alongside the abdomen (group B, n=50), and on a pillow ventrally to the chest with both arms flexed (group C, n=50). Two blinded, independent observers measured image noise and rated image quality (scores 1-3) of the liver, aorta, spleen, spine, and lower lungs. Radiation dose parameters were noted, and the abdomens' anterior-posterior diameter and scan lengths were measured. Interreader agreements for image noise (r=0.86; p<0.001) and subjective image quality (k=0.71-0.84) were good. Noise was lower (p<0.05), image quality of the liver, aorta, spleen, and spine was higher, and radiation dose lower in group A than in groups B and C (p<0.001, each). Image quality of the spleen, liver, and aorta were higher in group C than in group B (p<0.05, each). No significant differences in scan length (p=0.61) were found among groups. Abdominal anterior-posterior diameter correlated significantly with noise (r=0.82; p<0.01) and dose (r=0.47; p<0.001). Estimated effective radiation doses were significantly (p<0.001) higher in groups B (21.2 mSv) and C (21.9 mSv) as compared to A (16.1 mSv). In wbCT for polytrauma patients, positioning of the arms above the head results in better image quality and lower radiation dose. Placing the flexed arms on a large pillow ventrally to the chest significantly improves image quality as compared to positioning alongside the abdomen.


[3Fe-4S] <--> [4Fe-4S] cluster interconversion in Desulfovibrio africanus ferredoxin III: properties of an Asp14 --> Cys mutant.
The 8Fe ferredoxin III from Desulfovibrio africanus is a monomeric protein which contains two [4Fe-4S]2+/1+ clusters, one of which is labile and can readily and reversibly lose one Fe under oxidative conditions to yield a [3Fe-4S]1+/0 cluster. This 4Fe cluster has an S = 3/2 ground sping state insteaed of S = 1/2 in the reduced +1 state [George, Armstrong, Hatchikian and Thomson (1989) Biochem. J. 264, 275-284]. The co-ordination to this cluster is unusual in that an aspartate (Asp14, D14, is found where a cysteine residue normally occurs. Using a mutant protein obtained from the overexpression in Escherichia coli of a synthetic gene in which Asp14, the putative ligand to the removable Fe, has been changed to Cys, we have studied the cluster interconversion properties of the labile cluster. Analysis by EPR and magnetic-circular-dichroism spectroscopies showed that the Asp14 --> Cys (D14C) mutant contains two [4Fe-4S]2+/1+ clusters, both with S = 1/2 in the reduced state. Also, unlike in native 8Fe D. africanus ferredoxin III, the 4Fe <--> 3Fe cluster interconversion reaction was found to be sluggish and did not go to completion. It is inferred that the reversibility of the reaction in the native protein is due to the presence of the aspartate residue at position 14 and that this residue might protect the [3Fe-4S] cluster from further degradation.


Duplex Stem Replacement with bPNA+ Triplex Hybrid Stems Enables Reporting on Tertiary Interactions of Internal RNA Domains.
We report herein the synthesis and DNA/RNA binding properties of bPNA+, a new variant of bifacial peptide nucleic acid (bPNA) that binds oligo T/U nucleic acids to form triplex hybrids. By virtue of a new bivalent side chain on bPNA+, similar DNA affinity and hybrid thermostability can be obtained with half the molecular footprint of previously reported bPNA. Lysine derivatives bearing two melamine bases (K2M) can be prepared on multigram scale by double reductive alkylation with melamine acetaldehyde, resulting in a tertiary amine side chain that affords both peptide solubility and selective base-triple formation with 4 T/U bases; the Fmoc-K2M derivative can be used directly in solid phase peptide synthesis, rendering bPNA+ conveniently accessible. A compact bPNA+binding site of two U6 domains can be genetically encoded to replace existing 6 bp stem elements at virtually any location within an RNA transcript. We thus replaced internal 6 bp RNA stems that supported loop regions with 6 base-triple hybrid stems using fluorophore-labeled bPNA+. As the loop regions engaged in RNA tertiary interactions, the labeled hybrid stems provided a fluorescent readout; bPNA+ enabled this readout without covalent chemical modification or introduction of new structural elements. This strategy was demonstrated to be effective for reporting on widely observed RNA tertiary interactions such as intermolecular RNA-RNA kissing loop dimerization, RNA-protein binding, and intramolecular RNA tetraloop-tetraloop receptor binding, illustrating the potential general utility of this method. The modest 6 bp stem binding footprint of bPNA+ makes the hybrid stem replacement method practical for noncovalent installation of synthetic probes of RNA interactions. We anticipate that bPNA+ structural probes will be useful for the study of tertiary interactions in long noncoding RNAs.


Effect of Sugammadex on Postoperative Myasthenic Crisis in Myasthenia Gravis Patients: Propensity Score Analysis of a Japanese Nationwide Database.
In myasthenia gravis (MG) patients, postoperative myasthenic crisis, and residual neuromuscular blocking agent (NMBA) can cause respiratory failure that requires mechanical ventilation. However, it remains unclear whether the use of sugammadex for NMBA reversal reduces postoperative myasthenic crisis in MG patients undergoing surgery. We analyzed the association between use of sugammadex and postoperative myasthenic crisis in patients with MG using a national inpatient database. Adult patients with MG who received thymectomy under general anesthesia were identified in the Japanese Diagnosis Procedure Combination database from July 1, 2010 to March 31, 2016. Patients who received sugammadex (sugammadex group) were compared with those who did not receive sugammadex (control group). The primary outcome was postoperative myasthenic crisis, and the secondary outcomes were postoperative pneumonia, tracheostomy, 28-day mortality, total hospitalization costs, and length of stay after surgery. Propensity scores were estimated by logistic regression based on the following variables: age; sex; body mass index (BMI); smoking index; history of cancer; Charlson comorbidity index (CCI); type of thymectomy; time from hospital admission to surgery; use of plasma exchange, immunosuppressants, corticosteroids, anticholinesterase, and oral benzodiazepine before surgery; type of hospital; and treatment year. The outcomes were compared using stabilized inverse probability of treatment weighting (IPTW) analyses to obtain good between-group balance. Of 795 patients identified, 506 patients received sugammadex and 289 patients did not. After stabilized IPTW, the sugammadex group was associated with a decrease in postoperative myasthenic crisis (22/507 [4.3%] vs 25/288 [8.7%]; odds ratio [OR], 0.48; 95% confidence interval [CI], 0.25-0.91), but not associated with a decrease in postoperative pneumonia (5/507 [1.0%] vs 7/288 [2.4%]; OR, 0.44; 95% CI, 0.17-1.14) or tracheostomy (7/507 [1.4%] vs 10/288 [3.5%]; OR, 0.38; 95% CI, 0.12-1.22) compared with the control group. The sugammadex group had significantly lower median (interquartile range) total hospitalization costs ($13,186 [$11,250-$16,988] vs $14,119 [$11,713-$20,207]; P < .001) and median length of stay after surgery (10 [8-15] vs 11 [8-18] days; P < .001), compared with the control group. In this retrospective observational study, sugammadex was associated with reductions in postoperative myasthenic crisis and total hospitalization costs in adult patients with MG who received thymectomy. Given the present findings, sugammadex should be routinely administered for MG patients undergoing thymectomy.


Three dimensional magnetic abacus memory.
Stacking nonvolatile memory cells into a three-dimensional matrix represents a powerful solution for the future of magnetic memory. However, it is technologically challenging to access the data in the storage medium if large numbers of bits are stacked on top of each other. Here we introduce a new type of multilevel, nonvolatile magnetic memory concept, the magnetic abacus. Instead of storing information in individual magnetic layers, thereby having to read out each magnetic layer separately, the magnetic abacus adopts a new encoding scheme. It is inspired by the idea of second quantisation, dealing with the memory state of the entire stack simultaneously. Direct read operations are implemented by measuring the artificially engineered 'quantised' Hall voltage, each representing a count of the spin-up and spin-down layers in the stack. This new memory system further allows for both flexible scaling of the system and fast communication among cells. The magnetic abacus provides a promising approach for future nonvolatile 3D magnetic random access memory.


Fibre typing of intrafusal fibres.
The first descriptions of muscle spindles with intrafusal fibres containing striated myofibrils and nervous elements were given approximately 150 years ago. It took, however, another 100 years to establish the presence of two types of intrafusal muscle fibres: nuclear bag and nuclear chain fibres. The present paper highlights primarily the contribution of Robert Banks in fibre typing of intrafusal fibres: the confirmation of the principle of two types of nuclear bag fibres in mammalian spindles and the variation in occurrence of a dense M-band along the fibres. Furthermore, this paper summarizes how studies from the Umeå University group (Laboratory of Muscle Biology in the Department of Integrative Medical Biology) on fibre typing and the structure and composition of M-bands have contributed to the current understanding of muscle spindle complexity in adult humans as well as to muscle spindle development and effects of ageing. The variable molecular composition of the intrafusal sarcomeres with respect to myosin heavy chains and M-band proteins gives new perspectives on the role of the intrafusal myofibrils as stretch-activated sensors influencing tension/stiffness and signalling to nuclei.


Prenatal toxoplasmosis antibody and childhood autism.
There is evidence that some maternal infections during the prenatal period are associated with neurodevelopmental disorders, such as childhood autism. However, the association between autism and Toxoplasma gondii (T. gondii), an intracellular parasite, remains unclear. The authors examined whether serologically confirmed maternal antibodies to T. gondii are associated with odds of childhood autism in offspring. The study is based on a nested case-control design of a large national birth cohort (N = 1.2 million) and the national psychiatric registries in Finland. There were 874 cases of childhood autism and controls matched 1:1 on date of birth, sex, birthplace and residence in Finland. Maternal sera were prospectively assayed from a national biobank for T. gondii IgM and IgG antibodies; IgG avidity analyses were also performed. High maternal T. gondii IgM antibody was associated with a significantly decreased odds of childhood autism. Low maternal T. gondii IgG antibody was associated with increased offspring odds of autism. In women with high T. gondii IgM antibodies, the IgG avidity was high for both cases and controls, with the exception of three controls. The findings suggest that the relationship between maternal T. gondii antibodies and odds of childhood autism may be related to the immune response to this pathogen or the overall activation of the immune system. Autism Res 2017, 10: 769-777. © 2016 International Society for Autism Research, Wiley Periodicals, Inc.


Conformation and orientation of the retinyl chromophore in rhodopsin: a critical evaluation of recent NMR data on the basis of theoretical calculations results in a minimum energy structure consistent with all experimental data.
In the absence of a high-resolution diffraction structure, the orientation and conformation of the protonated Schiffs base retinylidinium chromophore of rhodopsin within the opsin matrix has been the subject of much speculation. There have been two recent reliable and precise NMR results that bear on this issue. One involves a determination of the C20-C10 and C20-C11 distances by Verdegem et al. [Biochemistry 38, 11316-11324 (1999)]. The other is the determination of the orientation of the methine C to methyl group vectors C5-C18, C9-C19, and C13-C20 relative to the membrane normal by Gröbner et al. [Nature 405 (6788), 810-813 (2000)]. Using molecular orbital methods that include extensive configuration interaction, we have determined what we propose to be the minimum energy conformation of this chromophore. The above NMR results permit us to check this structure in the C10-C11=C12-C13 region and then to check the global structure via the relative orientation of the three C18, C19, and C20 methyl groups. This method provides a detailed structure and also the orientation for the retinyl chromophore relative to the membrane normal and argues strongly that the protein does not appreciably alter the chromophore geometry from its minimum energy configuration that is nearly planar s-trans at the 6-7 bond. Finally, the chromophore structure and orientation presented in the recently published X-ray diffraction structure is compared with our proposed structure and with the deuterium NMR results.


Human papillomavirus types in women with invasive cervical carcinoma by HIV status in Kenya.
To evaluate the fraction of invasive cervical carcinoma (ICC) that could be prevented in HIV-infected women by vaccines currently available against human papillomavirus (HPV)16 and 18, we conducted a cross-sectional study in women with ICC in Nairobi, Kenya. Fifty-one HIV-positive women were frequency-matched by age to 153 HIV-negative women. Cervical cells were tested for HPV DNA using polymerase chain reaction-based assays (SPF10-INNO-LiPA). Comparisons were adjusted for multiplicity of HPV types. As expected, multiple-type infections were much more frequent in HIV-positive (37.2%) than in HIV-negative (13.7%) women, but the distribution of HPV types was similar. HPV16 was detected in 41.2% versus 43.8% and HPV16 and/or 18 in 64.7% versus 60.1% of HIV-positive versus HIV-negative women, respectively. The only differences of borderline statistical significance were an excess of HPV52 (19.6% versus 5.2%) and a lack of HPV45 (7.8% versus 17.0%) in HIV-positive women compared to HIV-negative women, respectively. We have been able to assess an unprecedented number of ICCs in HIV-positive women, but as we did not know the age of HIV acquisition, we cannot exclude that it had occurred too late in life to affect the type of HPV involved in cervical carcinogenesis. However, if our findings were confirmed, they would suggest that the efficacy of current vaccines against HPV16 and 18 to prevent ICC is similar in HIV-positive and HIV-negative women, provided vaccination is administered before sexual debut, as recommended.


Fiber optics for spectroscopic illumination.
Absolute and angular transmission measurements were made on a single sheathed fiber 0.15 mm in diameter and 2.1 m long, and an optical angular transfer function was defined and calculated. The effective transmission of the source brightness was calculated as a function of the f numbers of the entrance and output optics of the fiber, and these results were used to choose a suitable optical system for illumination of a spectrometer.


Patatin-like phospholipase domain-containing 3/adiponutrin deficiency in mice is not associated with fatty liver disease.
PNPLA3 (adiponutrin), a novel patatin-like phospholipase domain-containing enzyme, is expressed at high level in fat, but also in other tissues including liver. Polymorphisms in PNPLA3 have been linked to obesity and insulin sensitivity. Notably, a nonsynonymous variant rs738409(G) allele of the PNPLA3 gene was found to be strongly associated with both nonalcoholic and alcoholic fatty liver disease. We have generated Pnpla3(-/-) mice by gene targeting. Loss of Pnpla3 has no effect on body weight or composition, adipose mass, or development, whether the mice were fed regular chow or high-fat diet or bred into the genetic obese Lep(ob/ob) background. Plasma and liver triglyceride content and plasma aspartate aminotransferase and alanine aminotransferase levels were not different between Pnpla3(+/+) and Pnpla3(-/-) mice while they were on regular chow, fed three different fatty liver-inducing diets, or after they were bred into Lep(ob/ob) background. Hepatic Pnpla5 messenger RNA (mRNA) levels were similar in wild-type and Pnpla3(-/-) mice, although adipose Pnpla5 mRNA level was increased in Pnpla3(-/-) mice. A high-sucrose lipogenic diet stimulated hepatic Pnpla3 and Pnpla5 mRNA levels to a similar degree, but it did not affect adipose or liver triglyceride lipase (ATGL, known also as Pnpla2) mRNA in Pnpla3(+/+) and Pnpla3(-/-) mice. Finally, Pnpla3(+/+) and Pnpla3(-/-) mice displayed similar glucose tolerance and insulin tolerance tests while on regular chow or three different fatty liver-inducing diets. Loss of Pnpla3 does not cause fatty liver, liver enzyme elevation, or insulin resistance in mice.


A direct role for serum albumin in the cellular uptake of long-chain fatty acids.
The interaction of long-chain fatty acids with cells is important for their uptake and metabolism, as well as their involvement in signalling processes. The majority of long-chain fatty acids circulating in plasma exist as complexes with serum albumin. Thus an understanding of the involvement of serum albumin in these processes is vitally important. The effect of serum albumin on the uptake of long-chain fatty acids was studied in 3T3-L1 adipocytes. Serum albumin had a stimulatory effect on oleate uptake at all ratios of oleate: serum albumin tested. Furthermore, the rate of oleate uptake was saturable with increasing concentrations of serum albumin when the oleate: serum albumin ratio, and therefore the concentration of uncomplexed oleate, remained constant. This was not due to uptake being limited by dissociation of oleate from serum albumin, because oleate did not appear to be limiting. Furthermore, at very high ratios of oleate: serum albumin, when the concentration of uncomplexed oleate was predicted to be large relative to the amount of oleate taken up by cells, the rate of oleate uptake was still dependent on the albumin concentration. Serum albumin, covalently labelled with the photoreactive fatty acid 11-m-diazirinophenoxy[11-3H]undecanoate, bound to cells in a manner exhibiting both saturable (Kd 66.7 microM) and non-saturable processes. These results indicate that the stimulatory effect of serum albumin on the rate of oleate uptake is due to a direct interaction of serum albumin with the cells and point to an involvement of albumin binding sites in the cell surface in the cellular uptake of long-chain fatty acids.


Critical Review on Zeolite Clinoptilolite Safety and Medical Applications in vivo.
Unique and outstanding physical and chemical properties of zeolite materials make them extremely useful in a variety of applications including agronomy, ecology, manufacturing, and industrial processes. Recently, a more specific application of one naturally occurring zeolite material, clinoptilolite, has been widely studied in veterinary and human medicine. Due to a number of positive effects on health, including detoxification properties, the usage of clinoptilolite-based products in vivo has increased enormously. However, concerns have been raised in the public about the safety of clinoptilolite materials for in vivo applications. Here, we review the scientific literature on the health effects and safety in medical applications of different clinoptilolite-based materials and propose some comprehensive, scientifically-based hypotheses on possible biological mechanisms underlying the observed effects on the health and body homeostasis. We focus on the safety of the clinoptilolite material and the positive medical effects related to detoxification, immune response, and the general health status.


Peruvian mothers' knowledge and recognition of pneumonia in children under 5 years of age.
To assess Peruvian mothers' knowledge and recognition of pneumonia in children under 5 years of age, the mothers' attitude toward seeking medical help if they had a child with signs of pneumonia, and their perception of a Government of Peru pneumonia campaign. In this cross-sectional study, 501 mothers were selected randomly from 20 low-income communities of the metropolitan area of Lima, Peru, and were interviewed between June and August 2000. Using SPSS software, descriptive statistics were applied to summarize the demographic data and the data regarding the mothers' knowledge of pneumonia and recognition of signs of the disease. Cross-tabulations and chi-squares were done to assess relationships between variables and to make comparisons. About 84% of the mothers said that they knew what pneumonia is. Most believed that pneumonia is dangerous. A majority (58.7%) indicated that pneumonia is caused by lack of parental care. Only 28.9% believed that a virus causes the disease. More than 80% correctly picked rapid breathing and/or chest retraction from a list of possible signs and symptoms of pneumonia, and 94.6% said they were ready to take their child to the closest health center if they thought their child had pneumonia. Although 57.1% said they had heard about the Government of Peru pneumonia campaign, 69.3% of these mothers said they could not recall the motto of the campaign. Mothers who reported having heard of the campaign through TV were more likely than other mothers to correctly recognize the two major signs of pneumonia presented in the campaign. Although the percentage of mothers believing they can recognize pneumonia through rapid breathing and chest retraction seems to have increased in recent years, there is still a sizable percentage of mothers who remain uninformed about pneumonia and its possible fatal consequences. Efforts need to continue to educate Peruvian mothers about the causes, recognition of the signs, and treatment of pneumonia. The results suggest that the Government of Peru pneumonia campaign should use television much more, as well as the health centers, where most of the mothers receive medical attention and health information.


Chrysin, a natural and biologically active flavonoid suppresses tumor growth of mouse B16F10 melanoma cells: In vitro and In vivo study.
Chrysin (5,7-dihydroxyflavone) is a natural and biologically active compound which has many biological activities as an anticancer agent. The current report is aimed at finding out whether the antitumor potential of chrysin, evidenced in vitro and in vivo, is linked or not to its effect on immunological mechanisms of melanoma-bearing mice. Chrysin-treated B16F10 cells were analyzed for their metabolic rate and apoptotic potentials. In vivo, BALB/c mice received a subcutaneous injection of B16F10 melanoma cells prior to antitumor treatments with chrysin (50 mg/kg b.w) for 14 days and 21 days. The results showed that chrysin inhibited cancer cell growth at a dose-dependent manner by inducing apoptosis and cell cycle arrest at G2/M phase. Moreover, chrysin suppressed melanoma tumor growth at an average of 60% (after 14 days of treatment) and 71% (after 21 days of treatment) compared to the tumor-bearing group. Furthermore, chrysin treatment increased the cytotoxic activity of NK, CTL and macrophages. The findings showed that chrysin antitumor action on the murine melanoma model was very promising, suggesting that chrysin could be a potentially good candidate for future use in alternative anti-melanoma treatments.


Non-viral opportunistic infections in new users of tumour necrosis factor inhibitor therapy: results of the SAfety Assessment of Biologic ThERapy (SABER) study.
To determine among patients with autoimmune diseases in the USA whether the risk of non-viral opportunistic infections (OI) was increased among new users of tumour necrosis factor α inhibitors (TNFI), when compared to users of non-biological agents used for active disease. We identified new users of TNFI among cohorts of rheumatoid arthritis (RA), inflammatory bowel disease and psoriasis-psoriatic arthritis-ankylosing spondylitis patients during 1998-2007 using combined data from Kaiser Permanente Northern California, two pharmaceutical assistance programmes for the elderly, Tennessee Medicaid and US Medicaid/Medicare programmes. We compared incidence of non-viral OI among new TNFI users and patients initiating non-biological disease-modifying antirheumatic drugs (DMARD) overall and within each disease cohort. Cox regression models were used to compare propensity-score and steroid- adjusted OI incidence between new TNFI and non-biological DMARD users. Within a cohort of 33 324 new TNFI users we identified 80 non-viral OI, the most common of which was pneumocystosis (n=16). In the combined cohort, crude rates of non-viral OI among new users of TNFI compared to those initiating non-biological DMARD was 2.7 versus 1.7 per 1000-person-years (aHR 1.6, 95% CI 1.0 to 2.6). Baseline corticosteroid use was associated with non-viral OI (aHR 2.5, 95% CI 1.5 to 4.0). In the RA cohort, rates of non-viral OI among new users of infliximab were higher when compared to patients newly starting non-biological DMARD (aHR 2.6, 95% CI 1.2 to 5.6) or new etanercept users (aHR 2.9, 95% CI 1.5 to 5.4). In the USA, the rate of non-viral OI was higher among new users of TNFI with autoimmune diseases compared to non-biological DMARD users.


The hypoglossal canal and the origin of human vocal behavior.
The mammalian hypoglossal canal transmits the nerve that supplies the muscles of the tongue. This canal is absolutely and relatively larger in modern humans than it is in the African apes (Pan and Gorilla). We hypothesize that the human tongue is supplied more richly with motor nerves than are those of living apes and propose that canal size in fossil hominids may provide an indication about the motor coordination of the tongue and reflect the evolution of speech and language. Canals of gracile Australopithecus, and possibly Homo habilis, fall within the range of extant Pan and are significantly smaller than those of modern Homo. The canals of Neanderthals and an early "modern" Homo sapiens (Skhul 5), as well as of African and European middle Pleistocene Homo (Kabwe and Swanscombe), fall within the range of extant Homo and are significantly larger than those of Pan troglodytes. These anatomical findings suggest that the vocal capabilities of Neanderthals were the same as those of humans today. Furthermore, the vocal abilities of Australopithecus were not advanced significantly over those of chimpanzees whereas those of Homo may have been essentially modern by at least 400,000 years ago. Thus, human vocal abilities may have appeared much earlier in time than the first archaeological evidence for symbolic behavior.


Cellular biology of bone resorption.
Past knowledge and the recent developments on the formation, activation and mode of action of osteoclasts, with particular reference to the regulation of each individual step, have been reviewed. The following conclusions of consensus have emerged. 1. The resorption of bone is the result of successive steps that can be regulated individually. 2. Osteoclast progenitors are formed in bone marrow. This is followed by their vascular dissemination and the generation of resting preosteoclasts and osteoclasts in bone. 3. The exact pathways of differentiation of the osteoclast progenators to mature osteoclasts are debatable, but there is clear evidence that stromal cells support osteoclast generation. 4. Osteoclasts are activated following contact with mineralized bone. This appears to be controlled by osteoblasts that expose mineral to osteoclasts and/or release a factor that activates these cells. 5. Activated osteoclasts dissolve the bone mineral and digest the organic matter of bone by the action of agents secreted in the segregated microcompartments underlying their ruffled borders. The mineral is solubilized by protons generated from CO2 by carbonic anhydrase and secreted by an ATP-driven vacuolar H(+)-K(+)-ATPase located at the ruffled border. The organic matrix of the bone is removed by acid proteinases, particularly cysteine-proteinases that are secreted together with other lysosomal enzymes in the acid environment of the resorption zone. 6. Osteoclastic bone resorption is directly regulated by a polypeptide hormone, calcitonin (CT), and locally, by ionized calcium (Ca2+) generated as a result of osteoclastic bone resorption. 7. There is new evidence that osteoclast activity may also be influenced by the endothelial cells via generation of products including PG, NO and endothelin.


The Tyr-265-to-Cys mutator mutant of DNA polymerase beta induces a mutator phenotype in mouse LN12 cells.
DNA polymerase beta functions in both base excision repair and meiosis. Errors committed by polymerase beta during these processes could result in mutations. Using a complementation system, in which rat DNA polymerase beta substitutes for DNA polymerase I of Escherichia coli, we previously isolated a DNA polymerase beta mutant in which Tyr-265 was altered to Cys (Y265C). The Y265C mutant is dominant to wild-type DNA polymerase beta and possesses an intrinsic mutator activity. We now have expressed the wild-type DNA polymerase and the Y265C mutator mutant in mouse LN12 cells, which have endogenous DNA polymerase beta activity. We demonstrate that expression of the Y265C mutator mutant in the LN12 cells results in an 8-fold increase in the spontaneous mutation frequency of lambdacII mutants compared with expression of the wild-type protein. Expression of Y265C results in at least a 40-fold increase in the frequency of deletions of three bases or more and a 7-fold increase in point mutations. Our results suggest that the mutations we observe in vivo result directly from the action of the mutator polymerase. To our knowledge, this is the first demonstration of a mutator phenotype resulting from expression of a DNA polymerase mutator mutant in mammalian cells. This work raises the possibility that variant polymerases may act in a dominant fashion in human cells, leading to genetic instability and carcinogenesis.


Embryonic modulation of maternal steroids in European starlings (Sturnus vulgaris).
In birds, maternally derived yolk steroids are a proposed mechanism by which females can adjust individual offspring phenotype to prevailing conditions. However, when interests of mother and offspring differ, parent-offspring conflict will arise and embryonic interests, not those of the mother, should drive offspring response to maternal steroids in eggs. Because of this potential conflict, we investigated the ability of developing bird embryos to process maternally derived yolk steroids. We examined how progesterone, testosterone and oestradiol levels changed in both the yolk/albumen (YA) and the embryo of European starling eggs during the first 10 days of development. Next, we injected tritiated testosterone into eggs at oviposition to characterize potential metabolic pathways during development. Ether extractions separated organic and aqueous metabolites in both the embryo and YA homogenate, after which major steroid metabolites were identified. Results indicate that the concentrations of all three steroids declined during development in the YA homogenate. Exogenous testosterone was primarily metabolized to an aqueous form of etiocholanolone that remained in the YA. These results clearly demonstrate that embryos can modulate their local steroid environment, setting up the potential for parent-offspring conflict. Embryonic regulation must be considered when addressing the evolutionary consequences of maternal steroids in eggs.


Reliability and accuracy of MRI scanogram in the evaluation of limb length discrepancy.
The purpose of this study was to compare MRI scanogram with traditional radiographic methods for measurement of limb length. The authors hypothesized that MRI scanogram would be as reliable and accurate as radiographic scanogram in measurement of limb length without exposing patients to ionizing radiation. Twelve cadaveric femurs were measured using AP conventional radiographic scanogram, CT scanogram, MRI scanogram, and electronic caliper. Three orthopaedists performed two separate measurements using each technique. Intraobserver and interobserver variability was assessed for each of the three radiographic techniques. Accuracy was assessed by comparison of radiographic measurements to electronic caliper measurements of femur length. The reliability of all three radiographic limb length measurement techniques was excellent (ICC > 0.99). The accuracy of plain radiographic scanogram was slightly superior to CT scanogram and MRI scanogram. The mean absolute differences for radiographic, CT, and MRI scanograms compared with the gold standard, direct caliper measurement, were 0.52 mm, 0.68 mm, and 2.90 mm, respectively. All three scanogram techniques showed excellent reliability and accuracy. Radiographic scanogram remains the gold standard for leg length measurement. MRI scanogram is slightly less accurate compared with radiographic scanogram, but it does not use ionizing radiation. MRI scanogram merits clinical study and comparison with the traditional radiographic scanogram method for measurement of limb length.


Thermodynamic Separation of 1-Butene from 2-Butene in Metal-Organic Frameworks with Open Metal Sites.
Most C4 hydrocarbons are obtained as byproducts of ethylene production or oil refining, and complex and energy-intensive separation schemes are required for their isolation. Substantial industrial and academic effort has been expended to develop more cost-effective adsorbent- or membrane-based approaches to purify commodity chemicals such as 1,3-butadiene, isobutene, and 1-butene, but the very similar physical properties of these C4 hydrocarbons make this a challenging task. Here, we examine the adsorption behavior of 1-butene, cis-2-butene, and trans-2-butene in the metal-organic frameworks M2(dobdc) (M = Mn, Fe, Co, Ni; dobdc4- = 2,5-dioxidobenzene-1,4-dicarboxylate) and M2(m-dobdc) (m-dobdc4- = 4,6-dioxidobenzene-1,3-dicarboxylate), which all contain a high density of coordinatively unsaturated M2+ sites. We find that both Co2(m-dobdc) and Ni2(m-dobdc) are able to separate 1-butene from the 2-butene isomers, a critical industrial process that relies largely on energetically demanding cryogenic distillation. The origin of 1-butene selectivity is traced to the high charge density retained by the M2+ metal centers exposed within the M2(m-dobdc) structures, which results in a reversal of the cis-2-butene selectivity typically observed at framework open metal sites. Selectivity for 1-butene adsorption under multicomponent conditions is demonstrated for Ni2(m-dobdc) in both the gaseous and the liquid phases via breakthrough and batch adsorption experiments.


Psychological stress and cardiovascular disease: empirical demonstration of bias in a prospective observational study of Scottish men.
To examine the association between self perceived psychological stress and cardiovascular disease in a population where stress was not associated with social disadvantage. Prospective observational study with follow up of 21 years and repeat screening of half the cohort 5 years from baseline. Measures included perceived psychological stress, coronary risk factors, self reported angina, and ischaemia detected by electrocardiography. 27 workplaces in Scotland. 5606 men (mean age 48 years) at first screening and 2623 men at second screening with complete data on all measures. Prevalence of angina and ischaemia at baseline, odds ratio for incident angina and ischaemia at second screening, rate ratios for cause specific hospital admission, and hazard ratios for cause specific mortality. Both prevalence and incidence of angina increased with increasing perceived stress (fully adjusted odds ratio for incident angina, high versus low stress 2.66, 95% confidence interval 1.61 to 4.41; P for trend <0.001). Prevalence and incidence of ischaemia showed weak trends in the opposite direction. High stress was associated with a higher rate of admissions to hospital generally and for admissions related to cardiovascular disease and psychiatric disorders (fully adjusted rate ratios for any general hospital admission 1.13, 1.01 to 1.27, cardiovascular disease 1.20, 1.00 to 1.45, and psychiatric disorders 2.34, 1.41 to 3.91). High stress was not associated with increased admission for coronary heart disease (1.00, 0.76-1.32) and showed an inverse relation with all cause mortality, mortality from cardiovascular disease, and mortality from coronary heart disease, that was attenuated by adjustment for occupational class (fully adjusted hazard ratio for all cause mortality 0.94, 0.81 to 1.11, cardiovascular mortality 0.91, 0.78 to 1.06, and mortality from coronary heart disease 0.98, 0.75 to 1.27). The relation between higher stress, angina, and some categories of hospital admissions probably resulted from the tendency of participants reporting higher stress to also report more symptoms. The lack of a corresponding relation with objective indices of heart disease suggests that these symptoms did not reflect physical disease. The data suggest that associations between psychosocial measures and disease outcomes reported from some other studies may be spurious.


Crystal structure of human butyrylcholinesterase and of its complexes with substrate and products.
Cholinesterases are among the most efficient enzymes known. They are divided into two groups: acetylcholinesterase, involved in the hydrolysis of the neurotransmitter acetylcholine, and butyrylcholinesterase of unknown function. Several crystal structures of the former have shown that the active site is located at the bottom of a deep and narrow gorge, raising the question of how substrate and products enter and leave. Human butyrylcholinesterase (BChE) has attracted attention because it can hydrolyze toxic esters such as cocaine or scavenge organophosphorus pesticides and nerve agents. Here we report the crystal structures of several recombinant truncated human BChE complexes and conjugates and provide a description for mechanistically relevant non-productive substrate and product binding. As expected, the structure of BChE is similar to a previously published theoretical model of this enzyme and to the structure of Torpedo acetylcholinesterase. The main difference between the experimentally determined BChE structure and its model is found at the acyl binding pocket that is significantly bigger than expected. An electron density peak close to the catalytic Ser(198) has been modeled as bound butyrate.


Size-specific Dose Estimates for Chest, Abdominal, and Pelvic CT: Effect of Intrapatient Variability in Water-equivalent Diameter.
To develop software to automatically calculate size-specific dose estimates (SSDEs) and to assess the effect of variations in water-equivalent diameter (Dw) along the z-axis on SSDE for computed tomographic (CT) examinations of the torso. In this institutional review board-approved, HIPAA-compliant, retrospective study, a software program was used to calculate Dw at each image position in 102 consecutive CT examinations of the combined chest, abdomen, and pelvis. SSDE was calculated by multiplying the size-dependent conversion factor and volume CT dose index (CTDIvol) at each image position. The variations in Dw along the z-axis were determined for six hypothetical scanning ranges: chest alone; abdomen alone; pelvis alone; chest and abdomen; abdomen and pelvis; and chest, abdomen, and pelvis. Mean SSDE was calculated in two ways: (a) from the SSDE at each position and (b) from the mean CTDIvol over each scan range and the conversion factor corresponding to Dw at the middle of the scan range. Linear regression analysis was performed to determine the correlation between SSDE values calculated in these two ways. Across patients, for scan ranges 1-6, the mean of the difference between maximal and minimal Dw within a given patient was 5.2, 4.9, 2.5, 6.0, 5.6, and 6.5 cm, respectively. The mean SSDE values calculated by using the two methods were in close agreement, with root mean square differences of 0.9, 0.5, 0.5, 1.4, 1.0, and 1.1 mGy or 6%, 3%, 2%, 9%, 4%, and 6%, for the scan ranges of chest; abdomen; pelvis; chest and abdomen; abdomen and pelvis; and chest, abdomen, and pelvis, respectively. Using the mean CTDIvol from the whole scan range and Dw from the image at the center of the scan range provided an easily obtained estimate of SSDE for the whole scan range that agreed well with values from an image-by-image approach, with a root mean square difference less than 1.4 mGy (9%).


Ubiquitin system: direct effects join the signaling.
Ubiquitylation, a widespread and important posttranslational modification of eukaryotic proteins, regulates a multitude of critical cellular processes, both in normal and pathological conditions. A classical view of how ubiquitylation regulates protein function involves recognition of ubiquitin-encoded signals by specific ubiquitin-binding domains. However, evidence suggests the existence of direct effects of ubiquitylation, which occur through its impact on protein-protein interactions that do not involve specific ubiquitin receptors. Ubiquitin attachment may cause steric limitations that influence interaction of the modified protein with other proteins. Here, we present examples of this direct effect of ubiquitylation and propose how a two-level ubiquitin-mediated regulatory mechanism may provide flexibility.


Neonatal lung side population cells demonstrate endothelial potential and are altered in response to hyperoxia-induced lung simplification.
Lung side population (SP) cells are resident lung precursor cells with both epithelial and mesenchymal potential that are believed to play a role in normal lung development and repair. Neonatal hyperoxic exposure impairs lung development leading to a long-term decrease in gas exchange surfaces. The hypothesis that lung SP cells are altered during impaired lung development has not been studied. To address this issue, we characterized the endothelial potential of neonatal lung SP and subsets of lung SP from neonatal mice following hyperoxic exposure during room air recovery. Lung SP cells were isolated and sorted on the basis of their capacity to efflux Hoechst 33342. The lung SP was further sorted based on expression of Flk-1 and CD45. In vitro, both CD45(pos)/Flk-1(pos) and CD45(neg)/Flk-1(pos) bind isolectin B4 and incorporate LDL and form networks in matrigel, indicating that these populations have endothelial cell characteristics. Hyperoxic exposure of neonatal mice resulted in subtle changes in vascular and alveolar density on P13, which persisted with room air recovery to P41. During room air recovery, a decrease in lung SP cells was detected in the hyperoxic-exposed group on postnatal day 13 followed by an increase on day 41. Within this group, the lung SP subpopulation of cells expressing CD45 increased on day 21, 41, and 55. Here, we show that lung SP cells demonstrate endothelial potential and that the population distribution changes in number as well as composition following hyperoxic exposure. The hyperoxia-induced changes in lung SP cells may limit their ability to effectively contribute to tissue morphogenesis during room air recovery.


Randomized Phase III Study of Ganetespib, a Heat Shock Protein 90 Inhibitor, With Docetaxel Versus Docetaxel in Advanced Non-Small-Cell Lung Cancer (GALAXY-2).
Ganetespib, a highly potent heat shock protein 90 inhibitor, blocks multiple oncogenic pathways, resulting in antitumor activity. We evaluated the combination of ganetespib and docetaxel for second-line therapy of patients with advanced adenocarcinoma of the lung. In this international phase III trial, patients with stage IIIB or IV adenocarcinoma diagnosed > 6 months before study entry and 1 prior systemic therapy were randomly assigned (1:1) to ganetespib 150 mg/m2 on days 1 and 15 with docetaxel 75 mg/m2 on day 1 of a 21-day cycle or to docetaxel alone. The primary end point was overall survival (OS). Of 677 enrolled patients, 335 were randomly assigned to ganetespib and docetaxel and 337 were assigned to docetaxel. The trial was stopped early as a result of futility at a planned interim analysis. The median OS time was 10.9 months (95% CI, 9.0 to 12.3 months) in the ganetespib and docetaxel arm compared with 10.5 months (95% CI, 8.6 to 12.2 months) in docetaxel arm (hazard ratio [HR], 1.11; 95% CI, 0.899 to 1.372; P = .329). Median progression-free survival was 4.2 months in the ganetespib and docetaxel arm and 4.3 months in the docetaxel arm (HR, 1.16; 95% CI, 0.96 to 1.403; P = .119). The addition of ganetespib did not improve outcomes compared with docetaxel alone for any secondary end point, including survival in the elevated lactate dehydrogenase or EGFR and ALK wild-type populations. The most common grade 3 or 4 adverse event in both arms was neutropenia (30.9% with ganetespib and docetaxel v 25% with docetaxel). The addition of ganetespib to docetaxel did not result in improved survival for salvage therapy of patients with advanced-stage lung adenocarcinoma.


The crossbridge mechanism of muscular contraction and its implications.
The basic features of the sliding-filament crossbridge mechanism are reviewed briefly, and some recent objections involving supposed changes in A-filament lengths are discussed. X-ray diffraction studies on live muscles show no sign of a decrease in axial spacing during contraction, and it is unlikely that a stepwise shortening or depolymerization of A-filaments would provide a plausible contraction mechanism. Thus electron microscope observations which occasionally are reported to show such length changes probably arise from experimental artefact, of which there are many sources. The factors which govern tension and speed in muscle contraction are described. Since all vertebrate striated muscles which have been studied have A-bands of at least approximately the same length, they are likely to have rather similar maximum isometric tensions. The design probably matches this tension to the strength of the filaments themselves. The large variations in shortening speeds between different muscles and different animals arise because of corresponding variations in the rates of particular steps in the crossbridge cycle and in the rate of ATP splitting by the actin-myosin complex involved. Questions concerning the nature and the speed of the activation mechanism are also discussed.


M2 Kupffer cells promote M1 Kupffer cell apoptosis: a protective mechanism against alcoholic and nonalcoholic fatty liver disease.
Alcoholic and nonalcoholic fatty liver disease (ALD and NAFLD) are the predominant causes of liver-related mortality in Western countries. We have shown that limiting classical (M1) Kupffer cell (KC) polarization reduces alcohol-induced liver injury. Herein, we investigated whether favoring alternatively activated M2 KCs may protect against ALD and NAFLD. Ongoing alcohol drinkers and morbidly obese patients, with minimal hepatic injury and steatosis, displayed higher hepatic expression of M2 genes, as compared to patients with more severe liver lesions; individuals with limited liver lesions showed negligible hepatocyte apoptosis but significant macrophage apoptosis. Experiments in mouse models of ALD or NAFLD further showed that BALB/c or resveratrol-treated mice fed alcohol or a high-fat diet displayed preponderant M2 KC polarization, M1 KC apoptosis, and resistance to hepatocyte steatosis and apoptosis, as compared to control C57BL6/J mice. In vitro experiments in isolated KC, peritoneal, and Raw264.7 macrophages demonstrated that M1 macrophage apoptosis was promoted by conditioned medium from macrophages polarized into an M2 phenotype by either interleukin (IL)4, resveratrol, or adiponectin. Mechanistically, IL10 released from M2 cells promoted M1 death, and anti-IL10 antibodies blunted the proapoptic effects of M2-conditioned media. IL10 secreted by M2 KCs promoted selective M1 death by a mechanism involving activation of arginase in high inducible nitric oxide synthase-expressing M1 KCs. In alcohol-exposed mice, neutralization of IL10 impaired M1 apoptosis. These data uncover a novel mechanism regulating the M1/M2 balance that relies on apoptotic effects of M2 KCs towards their M1 counterparts. They suggest that promoting M2-induced M1 KC apoptosis might prove a relevant strategy to limit alcohol- and high fat-induced inflammation and hepatocyte injury.


Switching rates of multistep reactions.
We consider the switching rate of a metastable reaction scheme, which includes reactions with arbitrary steps, e.g., kA<-->(k+r)A (both forward and reverse reaction steps are allowed to happen). Employing a WKB approximation, controlled by a large system size, we evaluate both the exponent and the preexponential factor for the rate. The results are illustrated on a number of examples.


Conscious awareness is required for the perceptual discrimination of threatening animal stimuli: A visual masking and continuous flash suppression study.
We investigated if the subliminal processing of threatening animal (snakes and spiders) and neutral object (cars and houses) stimuli can influence the discrimination of a subsequent visible stimulus. The prime and target pair were either identical, of the same category but with different physical features, or different in category and physical features. In two experiments, participants discriminated the basic level category (e.g. snake vs. spider) of a visible target stimulus that had been preceded by a visible or perceptually invisible prime stimulus. One experiment used visual masking to render prime stimuli perceptually invisible and the other used continuous flash suppression (CFS). Priming effects were demonstrated in both experiments when the prime was visible but not when the prime was rendered perceptually invisible. These findings demonstrate that conscious awareness could be required in the perceptual discrimination of threatening animal and neutral object images at their specific basic level category.


Overweight adolescents eat what? And when? Analysis of consumption patterns to guide dietary message development for intervention.
Little is known about overweight adolescent dietary consumption patterns, with challenges to collecting meaningful data making it difficult to develop targeted obesity interventions. The present study aimed to examine the timing and consumption of fruit, vegetables and junk food by time of the day and day of the week. Overweight adolescents (n = 61), aged 12-16 years, completed 3-day food records. Negative binomial and binary logistic regression using generalised estimating equations were used to compare the amount and likelihood of the consumption of each food group between time periods. Overweight adolescent girls were more likely to eat fruit on weekdays than weekends [odds ratio (OR) = 5.0. P < 0.001], as were boys (OR = 2.5, P = 0.034). Adolescents consumed more fruit at school than other meals [girls: incident rate ratio (IRR) = 7.5, P < 0.001; boys: IRR = 4.0, P = 0.050]. Weekday dinner was the meal where girls were most likely to consume vegetables (OR = 3.0, P = 0.009) and when boys consumed the most vegetables (IRR = 30.9, P = 0.006). Fast food consumption was most likely for girls at dinner on the weekend (OR = 9.6, P = 0.042), whereas fast food intake for boys increased overall on the weekend (IRR = 3.6, P = 0.001). Intake of 'other junk' (e.g. crisps) peaked during school hours for girls (IRR = 7.2, P < 0.001) and sugar-sweetened beverage consumption increased for boys on the weekend (IRR = 3.3, P = 0.001). Overall, trends in fruit intake showed opposing times for high and low consumption compared to vegetable intake. These results represent the next step in using time of day and day of week consumption patterns to develop targeted, evidence-based dietary messages for interventions in overweight adolescents.


Anticancer activity of Nigella sativa (black seed) - a review.
Nigella sativa (N. sativa) seed has been an important nutritional flavoring agent and natural remedy for many ailments for centuries in ancient systems of medicine, e.g. Unani, Ayurveda, Chinese and Arabic Medicines. Many active components have been isolated from N. sativa, including thymoquinone, thymohydroquinone, dithymoquinone, thymol, carvacrol, nigellimine-N-oxide, nigellicine, nigellidine and alpha-hederin. In addition, quite a few pharmacological effects of N. sativa seed, its oil, various extracts and active components have been identified to include immune stimulation, anti-inflammation, hypoglycemic, antihypertensive, antiasthmatic, antimicrobial, antiparasitic, antioxidant and anticancer effects. Only a few authors have reviewed the medicinal properties of N. sativa and given some description of the anticancer effects. A literature search has revealed that a lot more studies have been recently carried out related to the anticancer activities of N. sativa and some of its active compounds, such as thymoquinone and alpha-hederin. Acute and chronic toxicity studies have recently confirmed the safety of N. sativa oil and its most abundant active component, thymoquinone, particularly when given orally. The present work is aimed at summarizing the extremely valuable work done by various investigators on the effects of N. sativa seed, its extracts and active principles against cancer. Those related to the underlying mechanism of action, derivatives of thymoquinone, nano thymoquinone and combinations of thymoquinone with the currently used cytotoxic drugs are of particular interest. We hope this review will encourage interested researchers to conduct further preclinical and clinical studies to evaluate the anticancer activities of N. sativa, its active constituents and their derivatives.


A Perspective on Accelerated Aging Caused by the Genetic Deficiency of the Metabolic Protein, OPA1.
Autosomal Dominant Optic Atrophy (ADOA) is an ophthalmological condition associated primarily with mutations in the OPA1 gene. It has variable onset, sometimes juvenile, but in other patients, the disease does not manifest until adult middle age despite the presence of a pathological mutation. Thus, individuals carrying mutations are considered healthy before the onset of clinical symptoms. Our research, nonetheless, indicates that on the cellular level pathology is evident from birth and mutant cells are different from controls. We argue that the adaptation and early recruitment of cytoprotective responses allows normal development and functioning but leads to an exhaustion of cellular reserves, leading to premature cellular aging, especially in neurons and skeletal muscle cells. The appearance of clinical symptoms, thus, indicates the overwhelming of natural cellular defenses and break-down of native protective mechanisms.


Mycobacterium tuberculosis Rv3628 drives Th1-type T cell immunity via TLR2-mediated activation of dendritic cells and displays vaccine potential against the hyper-virulent Beijing K strain.
Identification of vaccine target antigens (Ags) that induce Ag-specific Th1 immunity is the first step toward the development of a tuberculosis vaccine. Here, we evaluated the Mycobacterium tuberculosis (Mtb) protein Rv3628, a soluble inorganic pyrophosphatase, as a vaccine target and characterized the molecular details of its interaction with dendritic cells (DCs). Rv3628 activated DCs, increasing their expression of cell surface molecules and augmenting their production of TNF-α, IL-1β, IL-6, and IL-12p70. Rv3628 mediated these effects by binding to TLR2 and activating downstream MyD88-, MAPK- and NF-κB-dependent signaling pathways. Rv3628-stimulated DCs induced the expansion of OVA-specific CD4+ and CD8+ T cells, which secreted IFN-γ and IL-2. Rv3628-specific effector/memory T cells expanded to a similar extent as those stimulated with ESAT-6 Ag in samples of lung and spleen cells collected from Mtb-infected mice. Finally, an Rv3628 subunit vaccine adjuvanted with dimethyldioctadecylammonium liposomes containing monophosphoryl lipid-A caused significant reductions in bacterial counts and lung inflammation after challenge with the hyper-virulent Mtb K strain. Importantly, protective efficacy was correlated with the generation of Rv3628-specific CD4+ T cells co-producing IFN-γ, TNF-α and IL-2 and exhibiting an elevated IFN-γ recall response. Thus, Rv3628 polarizes DCs toward a Th1 phenotype and promotes protective immunity against Mtb infection.


Beliefs about optimal age and screening frequency predict breast screening adherence in a prospective study of female relatives from the Ontario site of the Breast Cancer Family Registry.
Although few studies have linked cognitive variables with adherence to mammography screening in women with family histories of breast and/or ovarian cancer, research studies suggest cognitive phenomena can be powerful adherence predictors. This prospective study included 858 women aged 30 to 71 years from the Ontario site of the Breast Cancer Family Registry with at least one first-degree relative diagnosed with breast and/or ovarian cancer. Data on beliefs about breast cancer screening and use of mammography were obtained from annual telephone interviews spanning three consecutive years. Self-reported mammogram dates were confirmed with medical imaging reports. Associations between beliefs about breast cancer screening and adherence with annual mammography were estimated using polytomous logistic regression models corrected for familial correlation. Models compared adherers (N = 329) with late-screeners (N = 382) and never-screeners (N = 147). Women who believed mammography screening should occur annually were more likely to adhere to annual screening recommendations than women who believed it should happen less often (OR: 5.02; 95% CI: 2.97-8.49 for adherers versus late-screeners; OR: 6.82; 95% CI: 3.29-14.16 for adherers versus never-screeners). Women who believed mammography screening should start at or before age 50 (rather than after) (OR: 9.72; 95% CI: 3.26-29.02) were significantly more likely to adhere when compared with never-screeners. Study results suggest that women with a family history of breast cancer should be strongly communicated recommendations about initial age of screening and screening intervals as related beliefs significantly predict adequate adherence.


Response of phenylketonuria to tetrahydrobiopterin.
A favorable response, indicated by decline of blood phenylalanine (Phe) in patients with phenylketonuria (PKU), to orally administered 6-R-L-erythro-5, 6, 7, 8-tetrahydrobiopterin (BH4) has been reported in many countries following the first publication in 1999. In this review, we describe the experience in the United States with PKU patients and their response to BH4. A significant response to BH4 is arbitrarily considered as a decrease of 30% or greater of blood Phe concentration 24 h after administration of BH4. In our studies, 18 of 37 patients with PKU (49%) responded to oral BH4 by >30% decrease in blood Phe concentration. Four PKU patients responded with a decrease of blood Phe concentration between 17.3 and 26.3%. It is suggested that patients with sufficient response to BH4 are candidates who will benefit from BH4 as it becomes available for PKU management. In a separate trial, 20 patients with PKU were screened with ascending doses of BH4: 10, 20, and 40 mg/kg. A favorable response was found in 10 subjects (50%) after 10 mg/kg BH4 and 14 subjects (70%) after 20 mg/kg BH4. There was no additional advantage to 40 mg/kg BH4. A 1-wk trial with 10 and 20 mg/kg BH4 in the same 20 patients showed blood Phe concentrations lowest after 7 d of BH4. The BH4-responsive patients were genotyped and most were compound heterozygotes with 1 mild mutation on 1 allele, responsible for the increase of the residual activity of Phe hydroxylase when BH4 was added. Individuals with the same genotype exhibit different responses upon administration of BH4, attributed to epigenetic factors, such as the metabolic makeup of the individual. Patients with PKU, regardless of their genotype or classification, need to be screened for response to BH4. The majority of patients are identified by 10 mg/kg BH4.