The genetics of Parkinson's disease: progress and therapeutic implications.
The past 15 years has witnessed tremendous progress in our understanding of the genetic basis for Parkinson's disease (PD). Notably, whereas most mutations, such as those in SNCA, PINK1, PARK2, PARK7, PLA2G6, FBXO7, and ATP13A2, are a rare cause of disease, one particular mutation in LRRK2 has been found to be common in certain populations. There has been considerable progress in finding risk loci. To date, approximately 16 such loci exist; notably, some of these overlap with the genes known to contain disease-causing mutations. The identification of risk alleles has relied mostly on the application of revolutionary technologies; likewise, second-generation sequencing methods have facilitated the identification of new mutations in PD. These methods will continue to provide novel insights into PD. The utility of genetics in therapeutics relies primarily on leveraging findings to understand the pathogenesis of PD. Much of the investigation into the biology underlying PD has used these findings to define a pathway, or pathways, to pathogenesis by trying to fit disparate genetic defects onto the same network. This work has had some success, particularly in the context of monogenic disease, and is beginning to provide clues about potential therapeutic targets. Approaches toward therapies are also being provided more directly by genetics, notably by the reduction and clearance of alpha-synuclein and inhibition of Lrrk2 kinase activity. We believe this has been an exciting, productive time for PD genetics and, furthermore, that genetics will continue to drive the etiologic understanding and etiology-based therapeutic approaches in this disease.


Osteoblast-specific factor 2: cloning of a putative bone adhesion protein with homology with the insect protein fasciclin I.
A cDNA library prepared from the mouse osteoblastic cell line MC3T3-E1 was screened for the presence of specifically expressed genes by employing a combined subtraction hybridization/differential screening approach. A cDNA was identified and sequenced which encodes a protein designated osteoblast-specific factor 2 (OSF-2) comprising 811 amino acids. OSF-2 has a typical signal sequence, followed by a cysteine-rich domain, a fourfold repeated domain and a C-terminal domain. The protein lacks a typical transmembrane region. The fourfold repeated domain of OSF-2 shows homology with the insect protein fasciclin I. RNA analyses revealed that OSF-2 is expressed in bone and to a lesser extent in lung, but not in other tissues. Mouse OSF-2 cDNA was subsequently used as a probe to clone the human counterpart. Mouse and human OSF-2 show a high amino acid sequence conservation except for the signal sequence and two regions in the C-terminal domain in which 'in-frame' insertions or deletions are observed, implying alternative splicing events. On the basis of the amino acid sequence homology with fasciclin I, we suggest that OSF-2 functions as a homophilic adhesion molecule in bone formation.


Subxiphoid thymectomy: single-port, dual-port, and robot-assisted.
Currently, surgical techniques that are less invasive than conventional median sternotomy are used for thymectomy in the treatment of myasthenia gravis and anterior mediastinal tumors as no sternal incision is required. We reported on a subxiphoid single-port thymectomy using CO2 insufflation, which has the following advantages: (I) the field of view offered by the camera scope inserted from the midline of the body helps confirm the location of the superior pole of the thymus and bilateral phrenic nerves; (II) there is minimal pain and no intercostal neuropathy occurs as intercostal spaces are not traversed; and (III) cosmetic outcomes are excellent. However, a drawback of this approach is that it requires familiarity with the single-port surgical procedure. Various surgical modifications have been suggested for the subxiphoid approach, which we currently use for thymectomy. These include subxiphoid single-port thymectomy; subxiphoid dual-port thymectomy (DPT) wherein an additional lateral thoracic intercostal port is added, which is used for more complicated surgeries; and subxiphoid robotic thymectomy using the da Vinci Surgical System. Here we report on these techniques. A subxiphoid approach in thymectomy is advantageous to patients as it minimizes or avoids the occurrence of intercostal neuropathy. Moreover, a subxiphoid approach provides the surgeon with a good field of view of the cervical region and helps confirm the location of the bilateral phrenic nerves. Therefore, thymectomy using a subxiphoid approach should be considered an option for a minimally invasive surgery.


Chromosomal localization of genes by scanning electron microscopy using in situ hybridization with biotinylated probes: Y chromosome repetitive sequences.
The feasibility of using scanning electron microscopy (SEM) to identify the position of specific DNA sequences was examined using a Y chromosome 'specific' probe (pHY2.1). Tests were carried out on chromosome spreads hybridized in situ with biotinylated pHY2.1. Chromosomal sites of hybridization of the probe were localized by an indirect immunohistochemical procedure which resulted in a gold product which could be amplified by silver precipitation. In the SEM, the specific location of the probe was easily identified due to the enhanced signal produced by the gold-silver complex. The probe was localized both on the long arm of the Y chromosome and within interphase nuclei. It was found that SEM was more sensitive than light microscopy since the probe could be identified without silver amplification. With refinements to the technique, SEM could provide a useful method for high resolution localizing of unique DNA sequences (i.e. single copy genes).


EZH2 is a potential therapeutic target for H3K27M-mutant pediatric gliomas.
Diffuse intrinsic pontine glioma (DIPG) is an aggressive brain tumor that is located in the pons and primarily affects children. Nearly 80% of DIPGs harbor mutations in histone H3 genes, wherein lysine 27 is substituted with methionine (H3K27M). H3K27M has been shown to inhibit polycomb repressive complex 2 (PRC2), a multiprotein complex responsible for the methylation of H3 at lysine 27 (H3K27me), by binding to its catalytic subunit EZH2. Although DIPGs with the H3K27M mutation show global loss of H3K27me3, several genes retain H3K27me3. Here we describe a mouse model of DIPG in which H3K27M potentiates tumorigenesis. Using this model and primary patient-derived DIPG cell lines, we show that H3K27M-expressing tumors require PRC2 for proliferation. Furthermore, we demonstrate that small-molecule EZH2 inhibitors abolish tumor cell growth through a mechanism that is dependent on the induction of the tumor-suppressor protein p16INK4A. Genome-wide enrichment analyses show that the genes that retain H3K27me3 in H3K27M cells are strong polycomb targets. Furthermore, we find a highly significant overlap between genes that retain H3K27me3 in the DIPG mouse model and in human primary DIPGs expressing H3K27M. Taken together, these results show that residual PRC2 activity is required for the proliferation of H3K27M-expressing DIPGs, and that inhibition of EZH2 is a potential therapeutic strategy for the treatment of these tumors.


Rituxan nanoconjugation prolongs drug/cell interaction and enables simultaneous depletion and enhanced Raman detection of lymphoma cells.
Antibody therapy is a type of immunotherapy that uses monoclonal antibodies (mAbs) to specifically bind to target cells to fight cancer. These mAbs have been conjugated with a number of nanocarriers to realize the targeted delivery of chemotherapeutics. Previous data indicate that the conjugation of a mAb to a nanocarrier promoted the faster internalization (endocytosis) of the nanoconjugates. In this study, we report that the crosslinking of Rituxan, an anti-lymphoma mAb, with silver nanoparticles prevented Rituxan from entering cells and prolonged drug/cell interaction. This finding was confirmed by both imaging flow cytometric and laser scanning confocal microscopic analyses of the fluorescently labeled nanoconjugates and cells. Furthermore, nanoconjugated Rituxan resulted in the enhanced capping of CD20 molecules on the cell membrane. We were able to show that the prolonged drug/cell interaction and enhanced capping directly contributed towards improved therapeutic efficiency. This represents a significant finding indicating that the nanocarrier served not only as a simple platform for targeted delivery, but also markedly altered the antibody performance at the molecular level and mediated key biological functions. The nanoconjugates also served as a Raman probe to specifically detect single live lymphoma cells with high sensitivity via surface-enhanced Raman scattering (SERS). Via integrating the single-cell level detection sensitivity of SERS with targeted and enhanced depletion ability, this nanoconjugate can be applied as a promising tool in lymphoma theranostics.


Chemical speciation of PM2.5 collected during prescribed fires of the Coconino National Forest near Flagstaff, Arizona.
The use of prescribed fire is expected to increase in an effort to reduce the risk of catastrophic fire, particularly at urban/forest interfaces. Fire is a well-known source of particulate matter (PM) with particle sizes < or =2.5 microm (PM2.5), small diameter PM known to affect climate, visibility, and human health. In this work, PM2.5 was collected during seven first-entry burns (flaming and smoldering stages) and one maintenance burn of the Coconino National Forest. Samples were analyzed for organic and elemental carbon, cations (sodium, potassium [K+], and ammonium [NH4+]), anions (nitrate [NO3-] and sulfate), and 48 elements (with atomic weights between sodium and lead). The PM2.5 contained high organic carbon levels (typically >90% by mass), commonly observed ions (K+, NH4+, and NO3-) and elements (K+, chlorine, sulfur, and silicon), as well as titanium and chromium. Flaming produced higher K+ and NH4+ levels than smoldering, and the elemental signature was more complex (20 versus 7 elements). Average organic carbon x 1.4 mass fractions (+/-standard deviation) were lower during flaming (92+/-14%) than during smoldering (124+/-24%). The maintenance (grassland) burn produced lower particle concentrations, lower NH4+ and NO3- levels, and higher K and chlorine levels than did the first-entry fires.


Top 100 Cited articles on Radiation Exposure in Medical Imaging: A Bibliometric Analysis.
Bibliometric analyses by highest number of citations can help researchers and funding agencies in determining the most influential articles in a field. The main objective of this analysis was to identify the top 100 cited articles addressing radiation exposure from medical imaging and assess their characteristics. Relevant articles were extracted from the Scopus database after a systematic search by researchers using an iteratively defined Boolean search string. Subsequently, exclusion criteria were applied. A list of top 100 articles was prepared, and articles were ranked according to the citations they had received. No time restriction was applied. Descriptive statistics of the data were compiled. The top-cited articles were published from 1970-2013, with the most articles published in 2009 and 2010 (12 articles in each year). The citations ranged from 107-1888 with a median of 272. Manuscripts from our top-cited list originated from 20 different countries, with contributions made by 158 authors and 160 organizations. Eighty-eight percent of studies evaluated patient-related radiation exposure, 7% health care workers, and 5% both or were not specified. Thirty-two percent of studies examined adult populations, 14% pediatric, and 54% included both populations or did not specify. Seventy-two percent of studies were dedicated to Computed Tomography, 8% to radiography/fluoroscopy, 9% to interventional procedures, 4% to nuclear medicine, and 7% to a combination of 2 or more modalities. The top 100 cited articles in medical imaging related to radiation exposure are diverse, originating from many countries with numerous contributing authors. The most common topics covered involve CT and adult patients. The recent peak in the most-highly cited articles (2010) suggests that increased attention has been devoted to this field in recent years. Based on these results, it would appear that research on radiation exposure in medical imaging is poised to continue expanding.


Plasmodium falciparum in vivo resistance to quinine: description of two RIII responses in French Guiana.
The resistance of Plasmodium falciparum to antimalarial drugs is one of the most worrisome problems in tropical medicine, but few clinical studies or observations have described confirmed cases of therapeutic failure. We report two cases of in vivo P. falciparum resistance (RIII response) to quinine in French Guiana, an Amazonian focal zone in which multi-resistant malaria is endemic. Both patients presented with uncomplicated malaria and were initially treated with intravenous quinine. Although absorption was normal, the treatment was not effective and the patients still had fever and significant parasitemia three days after the onset of treatment (day 3). The addition of intravenous tetracycline completely resolved the parasitemia within approximately 96 hours. These clinical reports confirm the necessity to combine quinine with tetracycline in this area, as recommended by the recent French regional antimalarial policy.


Age of greatest susceptibility to childhood lead exposure: a new statistical approach.
Susceptibility to lead toxicity is often assumed to be greatest during early childhood (e.g., 2 years of age), but recent studies suggest that blood lead concentrations (BPb) taken at 5-7 years of age are more strongly associated with IQ. We aimed to determine the age of greatest susceptibility to lead exposure using an innovative statistical approach that avoids the problem of correlated serial BPb measurements. We analyzed two cohorts of children that were followed from infancy to 6 years of age in Rochester, New York (n = 211), and Cincinnati, Ohio (n = 251). Serial BPb levels were measured and IQ tests were done when children were 6 years of age. After adjustment for relevant covariates, the ratio of 6-year BPb to 2-year BPb was added to the multiple regression model to test whether the pattern of BPb profiles during childhood had additional effect on IQ. The ratio of BPb at 6 years to the BPb at 2 years showed a strong effect on IQ (p < .001) when added to the multiple regression model that included the average childhood BPb. IQ decreased by 7.0 points for children whose BPb at 6 years of age was 50% greater than that at 2 years compared with children whose 6-year BPb was 50% less than their 2-year BPb. Similarly, criminal arrest rates were a factor of 3.35 higher for those subjects whose 6-year BPb was 50% higher than their 2-year BPb. We conclude that 6-year BPb is more strongly associated with cognitive and behavioral development than is BPb measured in early childhood.


Human Herpesviruses as Copathogens of HIV Infection, Their Role in HIV Transmission, and Disease Progression.
Of eight human herpesviruses (HHVs), often, only herpes simplex virus types 1 (HSV-1) and 2 (HSV-2) find mention in medical literature as both of these viruses are commonly associated with genital lesions and oral ulcers, commonly known as cold sores. However, role of human herpesviruses as copathogens and in aggravation and in the transmission of other human diseases, especially the Acquired immunodeficiency syndrome (HIV/AIDS) has only very recently been recognized. Therefore, screening and treating subclinical HHV infections may offer slowing of HIV infection, disease progression, and its transmission. Beside HSV-1 and HSV-2, HHV-3 a causative agent of herpes zoster remained one of the first manifestations of HIV disease before the era of highly active antiretroviral therapy (HAART). HHV-5 also known as human Cytomegalovirus infection remains a significant risk factor for HIV-associated mortality and morbidity even in HAART era. It is proposed that Cytomegalovirus viremia could be a better predictor of HIV disease progression than CD4+ T-lymphocyte count. The role of HHV-4 or Epstein-Burr virus and HHV-6, HHV-7, and HHV-8 is still being investigated in HIV disease progression. This review provides insight into the current understanding about these 8 HHVs, their co-pathogenesis, and role in HIV/AIDS disease progression. The review also covers recent literature in favor and against administering anti-HHV treatment along with HAART for slower AIDS progression and interrupted sexual transmission.


Clostridium Difficile Infection in Acute Care Hospitals: Systematic Review and Best Practices for Prevention.
OBJECTIVE Prevention of Clostridium difficile infection (CDI) in acute-care hospitals is a priority for hospitals and clinicians. We performed a qualitative systematic review to update the evidence on interventions to prevent CDI published since 2009. DESIGN We searched Ovid, MEDLINE, EMBASE, The Cochrane Library, CINAHL, the ISI Web of Knowledge, and grey literature databases from January 1, 2009 to August 1, 2015. SETTING We included studies performed in acute-care hospitals. PATIENTS OR PARTICIPANTS We included studies conducted on hospitalized patients that investigated the impact of specific interventions on CDI rates. INTERVENTIONS We used the QI-Minimum Quality Criteria Set (QI-MQCS) to assess the quality of included studies. Interventions were grouped thematically: environmental disinfection, antimicrobial stewardship, hand hygiene, chlorhexidine bathing, probiotics, bundled approaches, and others. A meta-analysis was performed when possible. RESULTS Of 3,236 articles screened, 261 met the criteria for full-text review and 46 studies were ultimately included. The average quality rating was 82% according to the QI-MQCS. The most effective interventions, resulting in a 45% to 85% reduction in CDI, included daily to twice daily disinfection of high-touch surfaces (including bed rails) and terminal cleaning of patient rooms with chlorine-based products. Bundled interventions and antimicrobial stewardship showed promise for reducing CDI rates. Chlorhexidine bathing and intensified hand-hygiene practices were not effective for reducing CDI rates. CONCLUSIONS Daily and terminal cleaning of patient rooms using chlorine-based products were most effective in reducing CDI rates in hospitals. Further studies are needed to identify the components of bundled interventions that reduce CDI rates. Infect Control Hosp Epidemiol 2017;38:476-482.


Low-income minority women at risk for cervical cancer: a process to improve adherence to follow-up recommendations.
The Houston Department of Health and Human Services (HDHHS) has a centralized process for notifying and setting up follow-up appointments for women with abnormal Pap smears who are clients of HDHHS health centers. Faculty and a student from the University of Texas School of Public Health and HDHHS personnel jointly conducted a study to evaluate the process and performance of the system. The study examined two subpopulations: women with atypical squamous cells of undetermined significance or low-grade squamous intraepithelial lesions (ASCUS/LGSIL) and women with high-grade squamous intraepithelial lesions or atypical glandular cells of undetermined significance (HGSIL/AGCUS). A retrospective study was conducted of data on women attending eight HDHHS clinics during the period from February 1996 through August 2000. Records of 1,216 women referred for evaluation of abnormal Pap smears were reviewed. Process effectiveness was measured by the number of successful contacts made and the number of appointments set up. Performance was measured by compliance with referral appointments. Predictors included race/ethnicity, age, co-existence of a sexually transmitted disease, number of prior referrals, type of patient visit, and health center attended. HDHHS staff successfully notified 95.6% of women with ASCUS/LGSIL and 97.9% of women with HGSIL/AGCUS. Using performance criteria as outcome measures, high-risk women requiring targeted interventions were identified. Overall, 84.2% of women scheduled appointments. Among those with ASCUS/LGSIL, women identified as African American were 53% less likely to accept an appointment and 45% less likely to show up for the appointment than those identified as Hispanic or "other." Age and type of patient visit appeared to be significantly associated with patient compliance behavior. The study describes the effectiveness of a centralized patient follow-up process for women at risk for cervical cancer.


The relationship between membrane fatty acid content and mitochondrial efficiency differs within- and between- omega-3 dietary treatments.
An important, but underappreciated, consequence of climate change is the reduction in crucial nutrient production at the base of the marine food chain: the long-chain omega-3 highly unsaturated fatty acids (n-3 HUFA). This can have dramatic consequences on consumers, such as fish as they have limited capacity to synthesise n-3 HUFA de novo. The n-3 HUFA, such as docosahexaenoic acid (DHA, 22:6n-3) and eicosapentaenoic acid (EPA, 20:5n-3), are critical for the structure and function of all biological membranes. There is increasing evidence that fish will be badly affected by reductions in n-3 HUFA dietary availability, however the underlying mechanisms remain obscure. Hypotheses for how mitochondrial function should change with dietary n-3 HUFA availability have generally ignored ATP production, despite its importance to a cell's total energetics capacity, and in turn, whole-animal performance. Here we (i) quantified individual variation in mitochondrial efficiency (ATP/O ratio) of muscle and (ii) examined its relationship with content in EPA and DHA in muscle membrane of a primary consumer fish, the golden grey mullet Chelon auratus, receiving either a high or low n-3 HUFA diet. Mitochondria of fish fed on the low n-3 HUFA diet had higher ATP/O ratio than those of fish maintained on the high n-3 HUFA diet. Yet, mitochondrial efficiency varied up about 2-fold among individuals on the same dietary treatment, resulting in some fish consuming half the oxygen and energy substrate to produce the similar amount of ATP than conspecific on similar diet. This variation in mitochondrial efficiency among individuals from the same diet treatment was related to individual differences in fatty acid composition of the membranes: a high ATP/O ratio was associated with a high content in EPA and DHA in biological membranes. Our results highlight the existence of interindividual differences in mitochondrial efficiency and its potential importance in explaining intraspecific variation in response to food chain changes.


Assessment of cognitive functions and adaptive behavior in children with end-stage renal disease on regular hemodialysis.
Chronic kidney disease (CKD) is defined by the Kidney Disease and Outcome Quality Initiative as a child who has kidney damage lasting for at least three months with or without decreased glomerular filtration rate. Hemodialysis (HD) means removal of waste products and extra fluid directly from the blood when the kidneys do not work properly. Studies aimed at investigating neurocognitive impairment in children with CKD have identified a wide range of delays in cognitive development. The aim of this study was to assess the cognitive functions and adaptive behavior in children with end-stage renal disease (ESRD) on regular HD. This case-controlled study was conducted on 30 children suffering from ESRD who were on treatment at the Pediatric Nephrology Unit of Tanta University Hospital. Thirty apparently healthy children served as a control group, in the period from January 2017 to January 2018. All children were subjected to full history taking, careful physical and neurological examination, specific investigations including assessment of intelligence quotient (IQ) using Stanford Binet test 5th edition; assessment of adaptive behavior, assessment of executive functions by using Wisconsin Card Sorting Test; and continuous performance test. This study showed that mean values of IQ and the Vineland test were significantly lower among patients than controls. The study suggests that children with ESRD had lower IQ, adaptive behavior and executive functions than healthy control children.


The ADP/ATP translocator is not essential for the mitochondrial permeability transition pore.
A sudden increase in permeability of the inner mitochondrial membrane, the so-called mitochondrial permeability transition, is a common feature of apoptosis and is mediated by the mitochondrial permeability transition pore (mtPTP). It is thought that the mtPTP is a protein complex formed by the voltage-dependent anion channel, members of the pro- and anti-apoptotic BAX-BCL2 protein family, cyclophilin D, and the adenine nucleotide (ADP/ATP) translocators (ANTs). The latter exchange mitochondrial ATP for cytosolic ADP and have been implicated in cell death. To investigate the role of the ANTs in the mtPTP, we genetically inactivated the two isoforms of ANT in mouse liver and analysed mtPTP activation in isolated mitochondria and the induction of cell death in hepatocytes. Mitochondria lacking ANT could still be induced to undergo permeability transition, resulting in release of cytochrome c. However, more Ca2+ than usual was required to activate the mtPTP, and the pore could no longer be regulated by ANT ligands. Moreover, hepatocytes without ANT remained competent to respond to various initiators of cell death. Therefore, ANTs are non-essential structural components of the mtPTP, although they do contribute to its regulation.


Alpha-Amylase Activity in Blood Increases after Pharmacological, But Not Psychological, Activation of the Adrenergic System.
Alpha-amylase in both blood and saliva has been used as a diagnostic parameter. While studies examining alpha-amylase activity in saliva have shown that it is sensitive to physiological and psychological challenge of the adrenergic system, no challenge studies have attempted to elucidate the role of the adrenergic system in alpha-amylase activity in blood. We set out to examine the impact of psychological and pharmacological challenge on alpha-amylase in blood in two separate studies. In study 1, healthy subjects were examined in a placebo-controlled, double-blind paradigm using yohimbine, an alpha2-adrenergic antagonist. In study 2, subjects were examined in a standardized rest-controlled psychosocial stress protocol. Alpha-amylase activity in blood was repeatedly measured in both studies. Results of study 1 showed that alpha-amylase in blood is subject to stronger increases after injection of yohimbine compared to placebo. In study 2, results showed that there was no significant effect of psychological stress compared to rest. Alpha-amylase in blood increases after pharmacological activation of the adrenergic pathways suggesting that sympathetic receptors are responsible for these changes. Psychological stress, however, does not seem to have an impact on alpha-amylase in blood. Our findings provide insight into the mechanisms underlying activity changes in alpha-amylase in blood in healthy individuals.


Feasibility of treatment of lower limb edema with calf muscle pump stimulation in chronic heart failure.
Persons with chronic heart failure may exhibit a decrease in functional ability related to lower extremity edema in spite of optimal diuretic therapy and salt restrictions. The aim of this pilot prospective clinical study was to test the feasibility of using exogenous calf muscle pump stimulation to decrease lower leg edema and thus improve functional status and quality of life. Six subjects entered into this study and agreed to use the intervention 30 min/day for one month. DXA was used to assess lower extremity composition. Device use averaged approximately 1 h/day and resulted in a reduction in the lean mass of the legs of 0.5 kg (range = 0.08-1.0 L; p = 0.03). Linear regression analysis of reduction of lower limb edema against daily usage suggests that increased utilization of calf muscle pump stimulation was associated with increased water losses, although this trend was not significant (R(2 = 0.4, p = 0.18). This pilot indicates that exogenous calf muscle pump stimulation could be a useful and safe addition to the patients' treatment regimes, but further studies testing a more typical population with heart failure is warranted.


Using the Neuroadaptagen KB200z™ to Ameliorate Terrifying, Lucid Nightmares in RDS Patients: the Role of Enhanced, Brain-Reward, Functional Connectivity and Dopaminergic Homeostasis.
Lucid Dreams are a form of dream life, during which the dreamer may be aware that he/she is dreaming, can stop/re-start the dreams, depending on the pleasantness or unpleasant nature of the dream, and experiences the dream as if he/she were fully awake. Depending on their content, they may be pleasant, un-pleasant or terrifying, at least in the context of patients, who also exhibit characteristics of Reward Deficiency Syndrome (RDS) and Posttraumatic Stress Disorder (PTSD). We present eight clinical cases, with known substance abuse, childhood abuse and diagnosed PTSD/RDS. The administration of a putative dopamine agonist, KB200Z™, was associated with the elimination of unpleasant and/or terrifying, lucid dreams in 87.5% of the cases presented, whereas one very heavy cocaine abuser showed a minimal response. These results required the continuous use of this nutraceutical. The lucid dreams themselves were distinguishable from typical, PTSD nightmares insofar as their content did not appear to reflect a symbolic rendition of an originally-experienced, historical trauma. Each of the cases was diagnosed with a form of RDS, i.e., ADHD, ADD, and/or Tourette's syndrome. They all also suffered from some form of Post-Traumatic-Stress-Disorder (PTSD) and other psychiatric diagnoses as well. The reduction or elimination of terrifying Lucid Dreams seemed to be dependent on KB220Z, whereby voluntary stopping of the agent results in reinstatement of the terrifying non-pleasant nature of the dreams. Following more required research on a much larger population we anticipate confirmation of these seemingly interesting observations. If these results in a small number of patients are indeed confirmed we may have found a frontline solution to a very perplexing and complicated symptom known as lucid dreams.


Impaired associative fear learning in mice with complete loss or haploinsufficiency of AMPA GluR1 receptors.
There is compelling evidence that l-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) glutamate receptors containing the GluR1 subunit contribute to the molecular mechanisms associated with learning. AMPA GluR1 glutamate receptor knockout mice (KO) exhibit abnormal hippocampal and amygdala plasticity, and deficits on various assays for cognition including Pavlovian fear conditioning. Here we examined associative fear learning in mice with complete absence (KO) or partial loss (heterozygous mutant, HET) of GluR1 on multiple fear conditioning paradigms. After multi-trial delay or trace conditioning, KO displayed impaired tone and context fear recall relative to WT, whereas HET were normal. After one-trial delay conditioning, both KO and HET showed impaired tone and context recall. HET and KO showed normal nociceptive sensitivity in the hot plate and tail flick tests. These data demonstrate that the complete absence of GluR1 subunit-containing receptors prevents the formation of associative fear memories, while GluR1 haploinsufficiency is sufficient to impair one-trial fear learning. These findings support growing evidence of a major role for GluR1-containing AMPA receptors in amygdala-mediated forms of learning and memory.


Intergenerational transmission of glucose intolerance and obesity by in utero undernutrition in mice.
Low birth weight (LBW) is associated with increased risk of obesity, diabetes, and cardiovascular disease during adult life. Moreover, this programmed disease risk can progress to subsequent generations. We previously described a mouse model of LBW, produced by maternal caloric undernutrition (UN) during late gestation. LBW offspring (F(1)-UN generation) develop progressive obesity and impaired glucose tolerance (IGT) with aging. We aimed to determine whether such metabolic phenotypes can be transmitted to subsequent generations in an experimental model, even in the absence of altered nutrition during the second pregnancy. We intercrossed female and male F(1) adult control (C) and UN mice and characterized metabolic phenotypes in F(2) offspring. We demonstrate that 1) reduced birth weight progresses to F(2) offspring through the paternal line (Cfemale -Cmale = 1.64 g; Cfemale -UNmale = 1.57 g, P < 0.05; UNfemale -Cmale = 1.64 g; UNfemale -UNmale = 1.60 g, P < 0.05), 2) obesity progresses through the maternal line (percent body fat: Cfemale -Cmale = 22.4%; Cfemale -UNmale = 22.9%; UNfemale -Cmale = 25.9%, P < 0.05; UNfemale -UNmale = 27.5%, P < 0.05), and 3) IGT progresses through both parental lineages (glucose tolerance test area under curve Cfemale -Cmale = 100; Cfemale -UNmale = 122, P < 0.05; UNfemale -Cmale = 131, P < 0.05; UNfemale -UNmale = 151, P < 0.05). Mechanistically, IGT in both F(1) and F(2) generations is linked to impaired beta-cell function, explained, in part, by dysregulation of Sur1 expression. Maternal undernutrition during pregnancy (F(0)) programs reduced birth weight, IGT, and obesity in both first- and second-generation offspring. Sex-specific transmission of phenotypes implicates complex mechanisms including alterations in the maternal metabolic environment (transmaternal inheritance of obesity), gene expression mediated by developmental and epigenetic pathways (transpaternal inheritance of LBW), or both (IGT).


The Better Assessment of Illness Study for Delirium Severity: Study Design, Procedures, and Cohort Description.
To describe the design, procedures, and cohort for the Better ASsessment of ILlness -(BASIL) study, which is conducted to develop and test new delirium severity measures, compare them with existing measures, and examine related clinical outcomes. Prospective cohort study with 1 year follow-up of study participants at a large teaching hospital in Boston, Massachusetts. After brief cognitive testing and the Delirium Symptom Interview, delirium and delirium severity were rated daily in the hospital using the Confusion Assessment Method (CAM) and CAM-Severity score, the Delirium Rating Scale-Revised-98 (DRS-R-98), and the Memorial Delirium Assessment Scale (MDAS). Other key study variables included comorbidity, physical function (basic and instrumental activities of daily living [ADL]), ratings of subjective health and well-being, and clinical outcomes (length of stay, 30 day rehospitalization, nursing home admission, healthcare utilization). Follow-up interviews occurred at 1- and 12-month with patients and families. In 42 patient interviews, inter-rater reliability for key variables was assessed. Of 768 eligible patients approached, 469 were screened and 352 enrolled, yielding an overall study response rate of 67% for potentially eligible participants. The mean participant was 80.3 years old (SD 6.8) and 203 (58%) were female. The majority of patients were medically complex with Charlson Comorbidity Scores ≥2 (192 patients, 55%), and 102 (29%) met criteria for dementia. Inter-rater reliability assessments (n = 42 pairs) were high for overall ratings of presence or absence of delirium by CAM (κ = 1.0), delirium severity by DRS-R-98 and MDAS (weighted kappa, κ = 1.0 for each) and for ADL impairment (κ = 1.0). For eligible participants at each time point, 278 out of 308 (90%) completed the 1-month follow-up and 132 out of 256 (53%) have completed the 12-month follow-up to date, which is still in progress. Among those who completed interviews, there was only 1-3% missing data on most major outcomes (delirium, basic ADL, and readmission). The BASIL study presents an innovative effort to advance the conceptualization and measurement of delirium severity. Unique strengths include the diverse cohort with complete high quality data and longitudinal follow-up, along with detailed collection of multiple delirium measures daily during hospitalization.


Shock waves on complex networks.
Power grids, road maps, and river streams are examples of infrastructural networks which are highly vulnerable to external perturbations. An abrupt local change of load (voltage, traffic density, or water level) might propagate in a cascading way and affect a significant fraction of the network. Almost discontinuous perturbations can be modeled by shock waves which can eventually interfere constructively and endanger the normal functionality of the infrastructure. We study their dynamics by solving the Burgers equation under random perturbations on several real and artificial directed graphs. Even for graphs with a narrow distribution of node properties (e.g., degree or betweenness), a steady state is reached exhibiting a heterogeneous load distribution, having a difference of one order of magnitude between the highest and average loads. Unexpectedly we find for the European power grid and for finite Watts-Strogatz networks a broad pronounced bimodal distribution for the loads. To identify the most vulnerable nodes, we introduce the concept of node-basin size, a purely topological property which we show to be strongly correlated to the average load of a node.


Evaluation of finger millet incorporated noodles for nutritive value and glycemic index.
The present study was undertaken to develop finger millet incorporated noodles for diabetic patients. Finger millet variety VL-149 was taken. The finger millet flour and refined wheat flour (RWF) were evaluated for nutrient composition. The finger millet flour (FMF) was blended in various proportions (30 to 50%) in refined wheat flour and used for the preparation of noodles. Control consisted of RWF noodles. Sensory quality and nutrient composition of finger millet noodles was evaluated. The 30% finger millet incorporated noodles were selected best on the basis of sensory evaluation. Noodles in that proportion along with control were evaluated for glycemic response. Nutrient composition of noodles showed that 50% finger millet incorporated noodles contained highest amount of crude fat (1.15%), total ash (1.40%), crude fiber (1.28%), carbohydrate (78.54%), physiological energy (351.36 kcal), insoluble dietary fiber (5.45%), soluble dietary fiber (3.71%), iron (5.58%) and calcium (88.39%), respectively. However, control RWF noodles contained highest amount of starch (63.02%), amylose (8.72%) and amylopectin (54.29%). The glycemic index (GI) of 30% finger millet incorporated noodles (best selected by sensory evaluation) was observed significantly lower (45.13) than control noodles (62.59). It was found that finger millet flour incorporated noodles were found nutritious and showed hypoglycemic effect.


Evaluation of an enzyme immunoassay for detection of dengue virus NS1 antigen in human serum.
We evaluated a one-step sandwich-format microplate enzyme immunoassay for detecting dengue virus NS1 antigen (Ag) in human serum by use of Platelia Dengue NS1 Ag kits (Bio-Rad Laboratories, Marnes La Coquette, France). We collected 299 serum samples from patients with dengue disease and 50 serum samples from patients not infected with dengue virus. For the 239 serum samples from patients with acute infections testing positive by reverse transcription-PCR and/or virus isolation for one of the four dengue virus serotypes, the sensitivity of the Platelia Dengue NS1 Ag kit was 88.7% (95% confidence interval, 84.0% to 92.4%). None of the serum samples from patients not infected with dengue virus tested positive with the Platelia Dengue NS1 Ag kit. A diagnostic strategy combining the Platelia Dengue NS1 Ag test for acute-phase sera and immunoglobulin M capture enzyme-linked immunosorbent assay for early-convalescent-phase sera increased sensitivity only from 88.7% to 91.9%. Thus, NS1 antigen detection with the Platelia Dengue NS1 Ag kit could be used for first-line testing for acute dengue virus infection in clinical diagnostic laboratories.


Blue light-excited LOV1 and LOV2 domains cooperatively regulate the kinase activity of full-length phototropin2 from Arabidopsis.
Phototropin2 (phot2) is a blue-light (BL) receptor that regulates BL-dependent activities for efficient photosynthesis in plants. phot2 comprises two BL-receiving light-oxygen-voltage-sensing domains (LOV1 and LOV2) and a kinase domain. BL-excited LOV2 is thought to be primarily responsible for the BL-dependent activation of the kinase. However, the molecular mechanisms by which small BL-induced conformational changes in the LOV2 domain are transmitted to the kinase remain unclear. Here, we used full-length wild-type and mutant phot2 proteins from Arabidopsis to study their molecular properties in the dark and under BL irradiation. Phosphorylation assays and absorption measurements indicated that the LOV1 domain assists the thermal relaxation of BL-excited LOV2 and vice versa. Using small-angle X-ray scattering and electron microscopy, we observed that phot2 forms a dimer and has a rod shape with a maximum length of 188 Å and a radius of gyration of 44 Å. Under BL, phot2 displayed large conformational changes that bent the rod shape. By superimposing the crystal structures of the LOV1 dimer, LOV2, and a homology model of the kinase to the observed changes, we inferred that the BL-dependent change consisted of positional shifts of both LOV2 and the kinase relative to LOV1. Furthermore, phot2 mutants lacking the photocycle in LOV1 or LOV2 still exhibited conformational changes under BL, suggesting that LOV1 and LOV2 cooperatively contribute to the conformational changes that activate the kinase. These results suggest that BL-activated LOV1 contributes to the kinase activity of phot2. We discuss the possible intramolecular interactions and signaling mechanisms in phot2.


A potent greenhouse gas identified in the atmosphere: SF(5)CF(3)
We detected a compound previously unreported in the atmosphere, trifluoromethyl sulfur pentafluoride (SF(5)CF(3)). Measurements of its infrared absorption cross section show SF(5)CF(3) to have a radiative forcing of 0.57 watt per square meter per parts per billion. This is the largest radiative forcing, on a per molecule basis, of any gas found in the atmosphere to date. Antarctic firn measurements show it to have grown from near zero in the late 1960s to about 0.12 part per trillion in 1999. It is presently growing by about 0.008 part per trillion per year, or 6% per year. Stratospheric profiles of SF(5)CF(3) suggest that it is long-lived in the atmosphere (on the order of 1000 years).


Evolutionary trends in the pharmacotherapeutic management of depression.
The past decade has brought an exponential increase in our knowledge of the pharmacotherapeutic management of depression. Much has been learned about the prevalence, risks, and course of depression in the general population, the elderly, and patients with comorbid medical illnesses and about the biological basis of depression. This article reviews evolving trends in the diagnosis and management of depression and evaluates the main classes of antidepressants. Although depression carries a high risk of morbidity and mortality, it is very treatable, and early diagnosis and early treatment are now emphasized. Antidepressant medication is continued after the patients' acute depressive symptoms resolve, sometimes for as long as 1 to 5 years to prevent relapse and recurrence of depression; in addition, full doses, rather than lower doses of antidepressant are prescribed for maintenance therapy. The armamentarium of antidepressants too has changed. In addition to monoamine oxidase inhibitors and tricyclic and tetracyclic antidepressants, the serotonin reuptake inhibitors fluoxetine, sertraline, and paroxetine are now available, as well as a group of antidepressants with atypical mechanisms of action that includes bupropion, trazodone, venlafaxin, and nefazodone. Although comparable in efficacy to the tricyclic antidepressants, these new drugs are safer and better tolerated because they are believed to act selectively on specific neurotransmitter systems.


Adaptation to sensory-motor reflex perturbations is blind to the source of errors.
In the study of visual-motor control, perhaps the most familiar findings involve adaptation to externally imposed movement errors. Theories of visual-motor adaptation based on optimal information processing suppose that the nervous system identifies the sources of errors to effect the most efficient adaptive response. We report two experiments using a novel perturbation based on stimulating a visually induced reflex in the reaching arm. Unlike adaptation to an external force, our method induces a perturbing reflex within the motor system itself, i.e., perturbing forces are self-generated. This novel method allows a test of the theory that error source information is used to generate an optimal adaptive response. If the self-generated source of the visually induced reflex perturbation is identified, the optimal response will be via reflex gain control. If the source is not identified, a compensatory force should be generated to counteract the reflex. Gain control is the optimal response to reflex perturbation, both because energy cost and movement errors are minimized. Energy is conserved because neither reflex-induced nor compensatory forces are generated. Precision is maximized because endpoint variance is proportional to force production. We find evidence against source-identified adaptation in both experiments, suggesting that sensory-motor information processing is not always optimal.


A mouse model for MeCP2 duplication syndrome: MeCP2 overexpression impairs learning and memory and synaptic transmission.
Rett syndrome and MECP2 duplication syndrome are neurodevelopmental disorders that arise from loss-of-function and gain-of-function alterations in methyl-CpG binding protein 2 (MeCP2) expression, respectively. Although there have been studies examining MeCP2 loss of function in animal models, there is limited information on MeCP2 overexpression in animal models. Here, we characterize a mouse line with MeCP2 overexpression restricted to neurons (Tau-Mecp2). This MeCP2 overexpression line shows motor coordination deficits, heightened anxiety, and impairments in learning and memory that are accompanied by deficits in long-term potentiation and short-term synaptic plasticity. Whole-cell voltage-clamp recordings of cultured hippocampal neurons from Tau-Mecp2 mice reveal augmented frequency of miniature EPSCs with no change in miniature IPSCs, indicating that overexpression of MeCP2 selectively impacts excitatory synapse function. Moreover, we show that alterations in transcriptional repression mechanisms underlie the synaptic phenotypes in hippocampal neurons from the Tau-Mecp2 mice. These results demonstrate that the Tau-Mecp2 mouse line recapitulates many key phenotypes of MECP2 duplication syndrome and support the use of these mice to further study this devastating disorder.


Clinical management practices adopted by physiotherapists in India for chronic obstructive pulmonary disease: A national survey.
Evidence supports the use of pulmonary rehabilitation in the treatment of chronic obstructive pulmonary disease (COPD) patients both during acute exacerbation and at later stages. It is used in India; but, to date, there has been no study that has investigated the structure of pulmonary rehabilitation programs for COPD patients in India. The recent study aims to determine the current practice patterns of Indian Physiotherapists for COPD patients admitted in Intensive Care Units (ICUs) and wards in terms of assessment and treatment. A questionnaire-based survey was conducted across India. Questionnaires were distributed to around 800 physiotherapists via E-mail. Physiotherapists with a Master Degree and a specialization in cardiopulmonary science or a minimum of 1 year of experience in treating cardiopulmonary patients were included. The questionnaires addressed assessment measures and treatment techniques given to COPD patients. A total of 342 completed questionnaires were received, yielding a response rate of 43.8%, with a majority of responses from Karnataka, Maharashtra and Gujarat. The assessment and treatment techniques used were almost similar between ICUs and wards. More than 80% of the responders carried out the assessment of certain respiratory impairments in both ICUs and wards. An objective measure of dyspnea was taken by less than 40% of the responders, with little attention given to functional exercise capacity and health-related quality of life. Eighty-five percent of the responders used Dyspnea-relieving strategies and traditional airway clearance techniques in both ICUs and wards. Eighty-three percent of the responders were giving patients in the wards training for upper and lower extremity. Fifty percent were giving strength training in the wards. Whether patients are admitted in ICUs or Wards, the practice pattern adopted by Physiotherapists to treat them vary very little with respect to certain measures taken. Assessment predominantly focused on respiratory impairment measures, followed by dyspnea-quantifying measures, with little attention given to functional exercise capacity and health-related quality of life measures. Treatment techniques given were concentrated on dyspnea-relieving strategies, airway clearance techniques and upper and lower extremity exercises, with little attention given to strength training.


Prioritizing Parental Worry Associated with Duchenne Muscular Dystrophy Using Best-Worst Scaling.
Duchenne muscular dystrophy (DMD) is a progressive, fatal pediatric disorder with significant burden on parents. Assessing disease impact can inform clinical interventions. Best-worst scaling (BWS) was used to elicit parental priorities among 16 short-term, DMD-related worries identified through community engagement. Respondents viewed 16 subsets of worries, identified using a balanced, incomplete block design, and identified the most and least worrying items. Priorities were assessed using best-worst scores (spanning +1 to -1) representing the relative number of times items were endorsed as most and least worrying. Independent-sample t-tests compared prioritization of parents with ambulatory and non-ambulatory children. Participants (n = 119) most prioritized worries about weakness progression (BW score = 0.64) and getting the right care over time (BW = 0.25). Compared to parents of non-ambulatory children, parents of ambulatory children more highly prioritized missing treatments (BW = 0.31 vs. 0.13, p < 0.001) and being a good enough parent (BW = 0.06 vs. -0.08, p = 0.010), and less prioritized child feeling like a burden (BW = -0.24 vs. -0.07, p < 0.001). Regardless of child's disease stage, caregiver interventions should address the emotional impact of caring for a child with a progressive, fatal disease. We demonstrate an accessible, clinically-relevant approach to prioritize disease impact using BWS, which offers an alternative to the use of traditional rating/ranking scales.


The Predictive Value of the Syntax Score in Patients With Chronic Coronary Artery Disease Undergoing Percutaneous Coronary Intervention or Coronary Artery Bypass Grafting: A Pilot Study.
To evaluate the usefulness of the SYNTAX score (SS) in predicting 1-year clinical outcomes in a population of patients with chronic coronary artery disease (CAD) undergoing percutaneous coronary intervention (PCI) or coronary artery bypass grafting (CABG). Despite the proven prognostic value of the SS in patients with multivessel and/or left main (LM) CAD, its usefulness in other patient subsets remains uncertain. This was a prospective single centre cohort study conducted from September 2012 to November 2014 at the Nicosia General Hospital, Cyprus. Patients (n=140; 94% men and 6% women) with chronic CAD undergoing revascularization with either PCI or CABG were evaluated. At 1-year, angina occurred in 20 patients (14.3%), myocardial infarction (MI) in 3 patients (2.1%), repeat revascularization procedures in 9 patients (6.4%) and death in 12 patients (8.6%). The SS independently predicted angina (p=0.024) but was not predictive of MI (p=0.964), death (p=0.292) or repeat revascularization (p=0.069). In this patient population, the SS predicted angina in the year following revascularization but was not predictive of MI, death or repeat revascularization.


Small molecule inhibitors of Hsp90 potently affect inflammatory disease pathways and exhibit activity in models of rheumatoid arthritis.
To evaluate the ability of SNX-7081, a novel small molecule inhibitor of Hsp90, to block components of inflammation, including cytokine production, protein kinase activity, and angiogenic signaling. A close analog was evaluated in preclinical in vivo models of rheumatoid arthritis (RA). SNX-7081 binding to Hsp90 was characterized in Jurkat cells and RA synovial fibroblasts (RASFs). Inhibition of NF-kappaB nuclear translocation was evaluated in cellular systems, using lipopolysaccharide (LPS), tumor necrosis factor alpha, or interleukin-1beta stimulation. Suppression of cytokine production in THP-1 cells, human umbilical vein endothelial cells, and RASFs was studied. Disruption of MAPK signaling cascades by SNX-7081 following growth factor stimulation was assessed. SNX-7081 was tested in 2 relevant angiogenesis assays: platelet-derived growth factor activation of fibroblasts and LPS-induced nitric oxide (NO) release in J774 macrophages. A close analog, SNX-4414, was evaluated in rat collagen-induced arthritis and adjuvant-induced arthritis, following oral treatment. SNX-7081 showed strong binding affinity to Hsp90 and expected induction of Hsp70. NF-kappaB nuclear translocation was blocked by SNX-7081 at nanomolar concentrations, and cytokine production was potently inhibited. Growth factor activation of ERK and JNK signaling was significantly reduced by SNX-7081. NO production was also sharply inhibited. In animal models, SNX-4414 fully inhibited paw swelling and improved body weight. Scores for inflammation, pannus formation, cartilage damage, and bone resorption returned to normal. The present results demonstrate that a small molecule Hsp90 inhibitor can impact inflammatory disease processes. The strong in vivo efficacy observed with SNX-4414 provides preclinical validation for consideration of Hsp90 inhibitors in the treatment of RA.


Extrinsic Repair of Injured Dendrites as a Paradigm for Regeneration by Fusion in Caenorhabditis elegans.
Injury triggers regeneration of axons and dendrites. Research has identified factors required for axonal regeneration outside the CNS, but little is known about regeneration triggered by dendrotomy. Here, we study neuronal plasticity triggered by dendrotomy and determine the fate of complex PVD arbors following laser surgery of dendrites. We find that severed primary dendrites grow toward each other and reconnect via branch fusion. Simultaneously, terminal branches lose self-avoidance and grow toward each other, meeting and fusing at the tips via an AFF-1-mediated process. Ectopic branch growth is identified as a step in the regeneration process required for bypassing the lesion site. Failure of reconnection to the severed dendrites results in degeneration of the distal end of the neuron. We discover pruning of excess branches via EFF-1 that acts to recover the original wild-type arborization pattern in a late stage of the process. In contrast, AFF-1 activity during dendritic auto-fusion is derived from the lateral seam cells and not autonomously from the PVD neuron. We propose a model in which AFF-1-vesicles derived from the epidermal seam cells fuse neuronal dendrites. Thus, EFF-1 and AFF-1 fusion proteins emerge as new players in neuronal arborization and maintenance of arbor connectivity following injury in Caenorhabditis elegans Our results demonstrate that there is a genetically determined multi-step pathway to repair broken dendrites in which EFF-1 and AFF-1 act on different steps of the pathway. EFF-1 is essential for dendritic pruning after injury and extrinsic AFF-1 mediates dendrite fusion to bypass injuries.


ZNF469 frequently mutated in the brittle cornea syndrome (BCS) is a single exon gene possibly regulating the expression of several extracellular matrix components.
Brittle cornea syndrome (BCS; MIM 229200) is an autosomal recessive generalized connective tissue disorder caused by mutations in ZNF469 and PRDM5. It is characterized by extreme thinning and fragility of the cornea that may rupture in the absence of significant trauma leading to blindness. Keratoconus or keratoglobus, high myopia, blue sclerae, hyperelasticity of the skin without excessive fragility, and hypermobility of the small joints are additional features of BCS. Transcriptional regulation of extracellular matrix components, particularly of fibrillar collagens, by PRDM5 and ZNF469 suggests that they might be part of the same pathway, the disruption of which is likely to cause the features of BCS. In the present study, we have performed molecular analysis of a cohort of 23 BCS affected patients on both ZNF469 and PRDM5, including those who were clinically reported previously [1]; the clinical description of three additional patients is reported in detail. We identified either homozygous or compound heterozygous mutations in ZNF469 in 18 patients while, 4 were found to be homozygous for PRDM5 mutations. In one single patient a mutation in neither ZNF469 nor PRDM5 was identified. Furthermore, we report the 12 novel ZNF469 variants identified in our patient cohort, and show evidence that ZNF469 is a single exon rather than a two exon gene.


Leakage-free polypyrrole-Au nanostructures for combined Raman detection and photothermal cancer therapy.
A bifunctional gold-coated polypyrrole (PPy-Au) nanostructure with Raman and photothermal activity was developed, in which polypyrrole spheres (PPy NSs) were used as the core and coated with small Au NPs to form polypyrrole-Au (PPy-Au) nanostructures. As a result of the contribution from the coated Au nanoparticles (Au NPs), coupling occurred between the Au NPs and the PPy NSs, leading to an enhanced pyrrole Raman signal. PPy-Au nanostructures were thus used as Raman tags without requiring additional Raman dyes labeling and leaking the cargo dyes even in the absence of a complicated encapsulation process. When conjugated with aptamer S2.2, the nanostructures could specifically bind with the membrane protein MUC1, a kind of tumor-related biomarker that is overexpressed on the surface of human breast cancer cells (MCF-7). The specific detection of MCF-7 cells was thus achieved based on the pyrrole Raman signal. Moreover, the capacity of the PPy-Au nanostructures for transforming NIR light into heat was also greatly enhanced because of the coupling between the Au NPs and the PPy NSs, as indicated by the efficiency of the photothermal conversion up to 70.0% compared to 44.7% for the pure PPy NSs. Photothermal experiments proved that MCF-7 cells incubated with the nanostructures nearly died under 808 nm irradiation (0.5 W cm-2) for 20 min, and the tumors bearing on the mice were also effectively inhibited after the PTT.


Regulation by heme of mitochondrial protein transport through a conserved amino acid motif.
A conserved motif, termed the heme regulatory motif (HRM), was identified in the presequences of the erythroid delta-aminolevulinate synthase precursors and was shown to be involved in hemin inhibition of transport of these proteins into mouse mitochondria in vitro. When the HRM was inserted into the presequence of the ornithine transcarbamoylase precursor, a normally unregulated mitochondrial protein, it conferred hemin inhibition on the transport of the chimeric protein. The conserved cysteine within the HRM was shown by site-directed mutagenesis to be required for hemin inhibition.


The epidemiology of chronic pain in the community.
Chronic pain is recognised as an important problem in the community but our understanding of the epidemiology of chronic pain remains limited. We undertook a study designed to quantify and describe the prevalence and distribution of chronic pain in the community. A random sample of 5036 patients, aged 25 and over, was drawn from 29 general practices in the Grampian region of the UK and surveyed by a postal self-completion questionnaire. The questionnaire included case-screening questions, a question on the cause of the pain, the chronic pain grade questionnaire, the level of expressed needs questionnaire, and sociodemographic questions. 3605 questionnaires were returned completed. 1817 (50.4%) of patients self reported chronic pain, equivalent to 46.5% of the general population. 576 reported back pain and 570 reported arthritis; these were the most common complaints and accounted for a third of all complaints. Backward stepwise logistic-regression modelling identified age, sex, housing tenure, and employment status as significant predictors of the presence of chronic pain in the community. 703 (48.7%) individuals with chronic pain had the least severe grade of pain, and 228 (15.8%) the most severe grade. Of those who reported chronic pain, 312 (17.2%) reported no expressed need, and 509 (28.0%) reported the highest expressed need. Chronic pain is a major problem in the community and certain groups within the population are more likely to have chronic pain. A detailed understanding of the epidemiology of chronic pain is essential for efficient management of chronic pain in primary care.


Susceptible stages in Schwann cells for NF1-associated plexiform neurofibroma development.
Stem cells are under strict regulation by both intrinsic factors and the microenvironment. There is increasing evidence that many cancers initiate through acquisition of genetic mutations (loss of intrinsic control) in stem cells or their progenitors, followed by alterations of the surrounding microenvironment (loss of extrinsic control). In neurofibromatosis type 1 (NF1), deregulation of Ras signaling results in development of multiple neurofibromas, complex tumors of the peripheral nerves. Neurofibromas arise from the Schwann cell lineage following loss of function at the NF1 locus, which initiates a cascade of interactions with other cell types in the microenvironment and additional cell autonomous modifications. In this study, we sought to identify whether a temporal "window of opportunity" exists during which cells of the Schwann cell lineage can give rise to neurofibromas following loss of NF1. We showed that acute loss of NF1 in both embryonic and adult Schwann cells can lead to neurofibroma formation. However, the embryonic period when Schwann cell precursors and immature Schwann cells are most abundant coincides with enhanced susceptibility to plexiform neurofibroma tumorigenesis. This model has important implications for understanding early cellular events that dictate neurofibroma development, as well as for the development of novel therapies targeting these tumors.


Surgery for T4 breast carcinoma: implications for local control.
Despite increasing public awareness and widespread availability of mammography, many patients will present with locally advanced breast cancers. The role of surgery remains controversial. Between 1993 and 1998, 47 of 393 (11.9%) breast cancer patients presented with T4 (inflammatory or locally advanced) carcinoma. We reviewed multimodality management, clinical response to neoadjuvant therapy, perioperative course and complications, and local control. Forty-six women and one man were diagnosed with clinical T4 breast cancer. There were 24 white and 23 African-American patients. Mean age at presentation was 54.5 (range, 31-88) years. Twenty-three patients had clinical metastases to axillary nodes, and five had distant metastases at the time of diagnosis. For these women, intent was for personal hygiene and control of pain. Neoadjuvant chemotherapy was given for 34 of 47 (72%) with 25 of 34 (73.5%) having partial or complete clinical response. There was no response or progression of disease in 9 of 34(26.5%). Forty-six patients underwent radical or modified radical mastectomy, whereas a single patient underwent breast conservation treatment. Twelve required tissue transfer for wound coverage. Although eight developed minor wound complications (cellulitis/flap separation), there were no major wound complications. Pathologically negative margins were achieved in all but one patient. To date, five women have developed local recurrence in either the chest wall (three) or axilla (two). Average time to local recurrence was 7.8 months. There is no evidence of local failure in the remaining 87 per cent. Locally advanced breast cancer is a common occurrence in certain populations. Multimodality management remains the standard of care. Surgical resection may allow for successful local control and, in certain situations, long-term cure.


Quality Use of the Pathology Data in Home Medicines Reviews: A Retrospective Evaluation.
Laboratory tests can be important tools for the assessment of pharmacotherapy. Nonetheless, there are no previous studies that have explicitly focused on the role of pathology data in Home Medicines Reviews (HMR), an Australian medication review program. Evaluate pharmacists' recommendations regarding laboratory testing in the medication review process. This retrospective review of HMRs assessed the prevalence of the pathology data provided by general practitioners. Additionally, the pharmacists' recommendations based on these laboratory data were compared with national and international guidelines. In total, 580 reports were evaluated. Of these, 179 reports did not contain any pathology data. Pharmacists commented on provided laboratory values in 324 reports and recommended further testing in 473 reports. Not all suggestions were related to previous values or were in line with guidelines. Most recommendations were regarding vitamin D and lipids (69% and 62% of medication review reports, respectively). Particularly, regarding renal impairment, pharmacists used their knowledge on dose adjustments and contraindications. In relation to full blood count, vitamin B12, and thyroid function, unjustified screenings were often recommended. In 26% of all reports, the pharmacists requested an array of tests without explaining the necessity for these tests. Conclusion and Relevance: Pharmacists provided useful advice based on the pathology data, which was concordant with national and international guidelines; however, in some cases, there was no rationale for the test recommendations provided. The outcome of the HMR program might be further enhanced if pharmacists had direct access to the patients' pathology data.


MESOMARK: a potential test for malignant pleural mesothelioma.
Soluble mesothelin-related peptides (SMRP)have been reported to be potential biomarkers for malignant pleural mesothelioma (MPM). We report analytical and preliminary clinical studies of MESOMARK, a quantitative assay for SMRP. The MESOMARK assay is a 2-step immunoenzymatic assay in an ELISA format with a 6-point calibration curve (0-32 nmol/L). We assessed analytical imprecision, analyte stability, and analytical interferences. We measured SMRP by this assay in 409 apparently healthy individuals (reference interval study), 177 patients with nonmalignant conditions, and 500 cancer patients, including 88 with MPM. The limit of detection was 0.16 nmol/L. At 2-19 nmol/L, intraassay imprecision (CV) was 1.1%-5.3%, and total imprecision was 4.0%-11.0%. The mean dilution recovery for 5 samples was 109% (range, 99%-113%). No interference was seen from added bilirubin (200 mg/L), hemoglobin (500 mg/L), triglycerides (30 g/L), chemotherapeutic agents, or other tested substances. Recombinant mesothelin was stable in serum upon freeze/thaw at -70 degrees C and upon storage for at least 7 days at 2-8 degrees C. The 99(th) percentile of the reference group was 1.5 nmol/L [95% confidence interval (CI), 1.2-1.6 nmol/L; n = 409], and mean SMRP was significantly higher in sera from patients with MPM (7.5 nmol/L; 95% CI, 2.8-12.1 nmol/L; n = 88). SMRP was increased in 52% and 5% of MPM patients and asbestos-exposed individuals, respectively. Concentrations in other nonmalignant and malignant conditions were similar to those in healthy controls. The MESOMARK assay is analytically robust and may be useful for the detection and management of mesothelioma.


Amerindian Ancestry Influences Genetic Susceptibility to Chronic Obstructive Pulmonary Disease.
The contribution of genetic ancestry on chronic obstructive pulmonary disease (COPD) predisposition remains unclear. To explore this relationship, we analyzed the associations between 754,159 single nucleotide polymorphisms (SNPs) and risk of COPD (n = 214 cases, 193 healthy controls) in Talca, Chile, considering the genetic ancestry and established risk factors. The proportion of Mapuche ancestry (PMA) was based on a panel of 45 Mapuche reference individuals. Five PRDM15 SNPs and two PPP1R12B SNPs were associate with COPD risk (p = 0.05 to 5×10-4) in those individuals with lower PMA. Based on linkage disequilibrium and sliding window analyses, an adjacent PRDM15 SNPs were associated with COPD risk in the lower PMA group (p = 10-3 to 3.77×10-8). Our study is the first to report an association between PPP1R12B and COPD risk, as well as effect modification between ethnicity and PRDM15 SNPs in determining COPD risk. Our results are biologically plausible given that PPP1R12B and PRDM15 are involved in immune dysfunction and autoimmunity, providing mechanistic evidence for COPD pathogenesis and highlighting the importance to conduct more genome wide association studies (GWAS) in admixed populations with Amerindian descent.


Ocular manifestations of the congenital varicella syndrome.
Three children developed the congenital varicella syndrome following a maternal varicella infection during the second trimester of pregnancy. Systemic findings included a bulbar palsy, mild hemiparesis, cicatricial skin lesions, developmental delay, and learning difficulties. Ocular findings included chorioretinitis, atrophy and hypoplasia of the optic discs, a congenital cataract, and Horner's syndrome. Electroretinograms and pattern-reversal visual-evoked potentials recorded from the four eyes with chorioretinitis were attenuated and degraded. Two of the children had been misdiagnosed as having congenital toxoplasmosis. Chorioretinal scars probably occur more frequently with the congenital varicella syndrome than has been previously recognized and should be included in the differential diagnosis of congenital chorioretinal scars. Hypoplasia of the optic disc may develop after an intrauterine varicella infection during the 16th week of gestation. Children with the congenital varicella syndrome should be examined by an ophthalmologist to exclude ocular abnormalities. These abnormalities are often subtle and difficult to diagnose correctly.


Human Papillomavirus Vaccination and Cervical Cytology Outcomes Among Urban Low-Income Minority Females.
The quadrivalent human papillomavirus (HPV) vaccine was licensed for use in 9- through 26-year-old females in 2006. Postlicensure studies in Australia, Denmark, and Canada have demonstrated vaccine effectiveness against abnormal cervical cytology results. However, there are limited data describing postlicensure effectiveness in the United States, particularly among minority females at higher risk for HPV infection and cervical cancer. To examine the effect of HPV vaccination on abnormal cervical cytology results among minority females. Retrospective cohort study conducted between January 2007 and January 2014 at 16 academically affiliated community clinics serving a low-income minority population. Included in this study was a population-based sample of 16 266 females aged 11 through 20 years as of January 1, 2007, who received care at a participating clinic on or after that date. Human papillomavirus vaccination, stratified by the number of doses. Cervical cytology abnormality following either HPV vaccination or, if unvaccinated, the first missed opportunity for HPV vaccination after January 1, 2007. Abnormalities were defined as atypical glandular cells, atypical squamous cells of undetermined significance, atypical squamous cells, cannot exclude a high-grade squamous intraepithelial lesion, low-grade squamous intraepithelial lesions, or high-grade squamous intraepithelial lesions. There were 4127 female patients who initiated quadrivalent HPV vaccination or had their first missed HPV vaccination opportunity from 11 through 20 years of age and underwent subsequent cervical cytology screening. The patients were primarily Spanish speaking (n = 2297; 58.3%) and publicly insured (n = 3801; 92.1%). The detection rate for an abnormal cervical cytology result during the observation period was lower among vaccinated (≥1 dose) (79.1 per 1000 person-years) vs unvaccinated (125.7 per 1000 person-years) females. The risk for an abnormal cervical cytology result was lower among vaccinated vs unvaccinated females (hazard ratio [HR], 0.64; 95% CI, 0.57-0.73), particularly if the 3-dose series was completed (HR, 0.48; 95% CI, 0.41-0.56) or if the vaccine was administered from 11 through 14 years of age (≥1 dose: HR, 0.36; 95% CI, 0.16-0.79; 3 doses: HR, 0.27; 95% CI, 0.12-0.63). This protective effect remained after adjusting for demographics, clinic type, abnormal baseline cervical cytology result, and baseline Chlamydia screening (as proxy for sexual experience). This study demonstrated the HPV vaccine is effective in a real-world setting of high-risk patients with variable HPV vaccination patterns.


[The winter projects grey from the eyes].
Selected texts derived from the edited diaries of an eighty year old man shed a light on the troubles of old age, declining health and wellbeing in a year prior to death. The author, Hans Warren (1921-2001) is a well known Dutch poet. Despite his poor physical condition and his mental frailty he confides his daily anguish and his lonely sufferings to his diary, resulting in an upsetting story of unhappiness and frustration. Although quite a few doctors are consulted, their advice seems inadequate or futile. The author's relationship with his supporting partner, who is nearly forty years his junior, suffers under the strain of daily conflicts and humiliating dependency. Despite all this they share moments of love and friendship until the end. The author expresses thankfulness for all things of beauty he can still enjoy.


Migration up and down the urban hierarchy and across the life course.
In this article, we begin by reviewing the concept of step migration that originated in E. G. Ravenstein's seminal papers "The Laws of Migration" (1885, 1889). As a result of the forces of the Industrial Revolution underway in 19th century Great Britain, migrants moved from farms to villages, from villages to towns, from towns to county seats, and thence to large cities. Throughout much of the industrialization era in the United States, net population movements similarly were upward within the urban hierarchy, and step migration today remains widespread throughout much of the still developing world. Our investigations of recent data and trends, however, suggest that the latest U.S. migration-pattern regime is a strongly contrasting one. Many of the major movements in the system of internal (or domestic) migration are flows down the urban hierarchy, although we note highly differentiated patterns for persons and households at specific stages of the life course. We make use of the newly defined metropolitan and micropolitan Core-Based Statistical Areas (CBSAs) and a seven-level size typology to tabulate origin-destination-specific migration flow data from both Census 2000 and IRS tax-return administrative records for the period 1995-2000. We discuss the causes for net movements being either upward or downward in the national urban hierarchy, including the effects of spatially focused immigration, and movement preferences at various ages, including migration in young adulthood associated with entering and leaving college and the military, as well as moves characteristic of the stages of family formation, childrearing, and retirement.


The role of the human papilloma virus in esophageal cancer.
Esophageal squamous cell carcinoma (ESCC) demonstrates wide regional variation in incidence and causal associations. Human papillomavirus (HPV) has been implicated in ESCC, particularly the sub-types 16 and 18. Transforming proteins E6 and E7 from these high risk sub-types, interact with p53 protein and Rb protein respectively, leading to loss of function of these tumor suppressor gene products. These interactions further lead to inactivation of the growth suppressive effects of the p53 and Rb proteins, resulting in abnormal proliferative states. p53 protein expression has been found in both HPV-positive and -negative tumors, indicating that HPV and p53 protein expression are not mutually exclusive and can occur together in the same tumor. It has been observed that HPV plays a more significant role in esophageal carcinogenesis in geographic areas with a high prevalence of the disease. A variation in the association between HPV and ESCC worldwide may be due to environmental and geographic factors, or to genetic susceptibility to esophageal HPV infections. Variations in the sensitivity of techniques used in the detection of the virus and in the methodology for processing the tumor tissues, may also be responsible for global differences. Esophageal carcinogenesis is a complex multistep process with a multifactorial etiology. Infection with oncogenic HPV types may be an integral part in a multistep process that leads to ESCC.


Rest-activity rhythms in emerging adults: implications for cardiometabolic health.
Emerging adulthood (18-25 years) represents a window of opportunity to modify the trajectory of cardiometabolic disease risk into older adulthood. Not known is the extent to which rest-activity rhythms (RAR) may be related to biomarkers of cardiometabolic health in this population. In this cross-sectional, observational study, 52 healthy emerging adults wore wrist accelerometers (14 consecutive days; 24 h/day) for assessment of nonparametric RAR metrics, including interdaily stability (IS; day-to-day RAR consistency), intradaily variability (IV; within-day RAR fragmentation), and relative amplitude (RA; robustness of RAR), as well as autocorrelation (correlation of rest/activity levels at 24-h lag-times). Cardiometabolic biomarkers, including body mass index (BMI), body fat percentage, blood pressure (BP), fasting lipids, glucose, and C-reactive protein (CRP) were assessed. Additional measures including physical activity, sleep duration, and habitual caffeine and alcohol consumption were also evaluated. A series of multivariable regression models of cardiometabolic biomarkers were used to quantify associations with RAR metrics. On average, participants were 20 ± 1 years of age (21 males, 31 females), non-obese, and non-hypertensive. All were nonsmokers and free of major diseases or conditions. In separate models, which adjusted for sex, BMI, moderate-vigorous physical activity, sleep duration, caffeine, and alcohol consumption, IS was inversely associated with total cholesterol (p ≤ 0.01) and non-HDL cholesterol (p < .05), IV was positively associated with CRP (p < .05), and autocorrelation was inversely associated with total cholesterol (p < .05) and CRP (p < .05). Conversely, associations between RA and cardiometabolic biomarkers were nonsignificant after adjustment for BMI, alcohol, and caffeine consumption. In conclusion, RAR metrics, namely, a higher IS, lower IV, and higher autocorrelation, emerged as novel biomarkers associated with more favorable indices of cardiometabolic health in this sample of apparently healthy emerging adults.


Brain volume change and cognitive trajectories in aging.
Examine how longitudinal cognitive trajectories relate to brain baseline measures and change in lobar volumes in a racially/ethnically and cognitively diverse sample of older adults. Participants were 460 older adults enrolled in a longitudinal aging study. Cognitive outcomes were measures of episodic memory, semantic memory, executive function, and spatial ability derived from the Spanish and English Neuropsychological Assessment Scales (SENAS). Latent variable multilevel modeling of the four cognitive outcomes as parallel longitudinal processes identified intercepts for each outcome and a second order global change factor explaining covariance among the highly correlated slopes. We examined how baseline brain volumes (lobar gray matter, hippocampus, and white matter hyperintensity) and change in brain volumes (lobar gray matter) were associated with cognitive intercepts and global cognitive change. Lobar volumes were dissociated into global and specific components using latent variable methods. Cognitive change was most strongly associated with brain gray matter volume change, with strong independent effects of global gray matter change and specific temporal lobe gray matter change. Baseline white matter hyperintensity and hippocampal volumes had significant incremental effects on cognitive decline beyond gray matter change. Baseline lobar gray matter was related to cognitive decline, but did not contribute beyond gray matter change. Cognitive decline was strongly influenced by gray matter volume change and, especially, temporal lobe change. The strong influence of temporal lobe gray matter change on cognitive decline may reflect involvement of temporal lobe structures that are critical for late life cognitive health but also are vulnerable to diseases of aging. (PsycINFO Database Record


BIOLOGICAL FORMATION OF MOLECULAR HYDROGEN.
From a general standpoint, the formation of molecular hydrogen can be considered a device for disposal of electrons released in metabolic oxidations. We presume that this means of performing anaerobic oxidations is of ancient origin and that the hydrogen-evolving system of strict anaerobes represents a primitive form of cytochrome oxidase, which in aerobes effects the terminal step of respiration, namely the disposal of electrons by combination with molecular oxygen. We further assume that the original pattern of reactions leading to H(2) production has become modified in various ways (with respect to both mechanisms and functions) during the course of biochemical evolution, and we believe that this point of view suggests profitable approaches for clarifying a number of problems in the intermediary metabolism of microorganisms which produce or utilize H(2). Of special general importance in this connection is the basic problem of defining more precisely the fundamental elements in the regulatory control of anaerobic energy metabolism. Among the more specific aspects awaiting further elucidation are: the relations between formation of H(2) and use of H(2) as a primary reductant for biosynthetic purposes; the various forms of direct and indirect interactions between hydrogenase and N(2) reduction systems; and the transitional stages between anaerobic and aerobic energy-metabolism patterns of facultative organisms.


[Evaluation of coverage by a neonatal screening program for hemoglobinopathies in the Recôncavo region of Bahia, Brazil].
In 2001, government ruling n. MS 822/01 by the Brazilian Ministry of Health made neonatal screening mandatory for hemoglobinopathies, with special focus on sickle cell disease, the most common hemoglobinopathy in Brazil. Bahia is the State of Brazil with the highest prevalence of this disease. The current paper reports on the prevalence of hemoglobin variants HbS and HbC, which cause sickle cell disease, in the cities of Cachoeira, Maragogipe, and São Félix, Bahia State. The overall proportion of carriers for the two forms of hemoglobin was 13%. From 2001 to 2003, the neonatal screening rate decreased from 82.6% to 46.4% in Cachoeira and increased from 37.0% to 56.2% in Maragogipe. Thus, only about one-half of children are being tested in these cities. The findings show that the public health care system in these cities is poorly prepared to screen for sickle cell disease in newborns and that there was a lack of health care personnel to follow up on newly diagnosed sickle cell patients or carriers of the sickle cell trait.


Receptor for activated protein kinase C1 regulates cell proliferation by modulating calcium signaling.
Receptor for activated protein kinase C1 (RACK1) is an intracellular scaffolding protein known to interact with the inositol-1,4,5-trisphosphate receptor and thereby enhance calcium release from the sarcoplasmic reticulum. Because calcium signaling may affect vascular smooth muscle cell proliferation, we investigated whether RACK1 regulates proliferation of rat preglomerular microvascular smooth muscle cells. Western blot analysis indicated that preglomerular microvascular smooth muscle cells robustly express RACK1 protein, and coimmunoprecipitation experiments demonstrated that RACK1 binds the inositol-1,4,5-trisphosphate receptor. RACK1 small interfering RNA (siRNA) decreased RACK1 mRNA and protein expression, significantly (P=0.0225) reduced steady-state basal levels of intracellular calcium (6712±156 versus 7408±248, arbitrary fluorescence units in RACK1 siRNA-treated versus control cells, respectively) and significantly (P<0.0001) decreased cell proliferation by ≈50%. Xestospongin C and 2-aminoethoxydiphenyl borate (antagonists of inositol-1,4,5-trisphosphate receptors), cyclopiazonic acid (sarcoplasmic reticulum Ca(2+)-ATPase inhibitor), and calmidazolium (calmodulin inhibitor) mimicked the effects of RACK1 siRNA on proliferation, and RACK1 siRNA had no additional effects on proliferation in the presence of these agents. RACK1 siRNA did not affect the expression of cyclin D1/2 or phosphorylation of retinoblastoma protein (progrowth cell cycle regulators), yet it caused compensatory decreases in the expression of p21(Cip1/Waf1) and p27(Kip1) (antigrowth cell cycle regulators). Like preglomerular microvascular smooth muscle cells, glomerular mesangial cells also expressed high levels of RACK1, and RACK1 siRNA inhibited their proliferation. In conclusion, RACK1 modulates proliferation of preglomerular microvascular smooth muscle cells and glomerular mesangial cells, likely via the inositol-1,4,5-trisphosphate receptor/calcium/calmodulin pathway. RACK1 may represent a novel druggable target for treating renal diseases, such as glomerulosclerosis.


Cardiovascular death in rheumatoid arthritis: a population-based study.
To determine whether systemic inflammation confers any additional risk for cardiovascular death among patients with rheumatoid arthritis (RA), after adjusting for traditional cardiovascular risk factors and comorbidities. Using the population-based data resources of the Rochester Epidemiology Project, we assembled an incidence cohort of all Rochester, Minnesota residents ages >or=18 years who first fulfilled the American College of Rheumatology 1987 criteria for RA between January 1, 1955 and January 1, 1995. All subjects were followed up longitudinally through their complete (inpatient, outpatient) medical records, beginning at age 18 years and continuing until death, migration, or January 1, 2001. Detailed information on the occurrence of various cardiovascular risk factors (personal history of coronary heart disease [CHD], congestive heart failure, smoking, hypertension, dyslipidemia, body mass index [BMI], diabetes mellitus, menopausal status) as well as indicators of systemic inflammation and RA disease severity (rheumatoid factor [RF] seropositivity, erythrocyte sedimentation rate [ESR], joint swelling, radiographic changes, RA nodules, RA complications, RA treatments, disease duration) and comorbidities were collected on all subjects. Causes of death were ascertained from death certificates and medical records. Cox regression models were used to estimate the independent predictors of cardiovascular death. This inception cohort comprised a total of 603 RA patients whose mean age was 58 years, of whom 73% were women. During a mean followup of 15 years, 354 patients died and cardiovascular disease was the primary cause of death in 176 patients. Personal history of CHD, smoking, hypertension, low BMI, and diabetes mellitus, as well as comorbidities, including peripheral vascular disease, cerebrovascular disease, chronic pulmonary disease, dementia, ulcers, malignancies, renal disease, liver disease, and history of alcoholism, were all significant risk factors for cardiovascular death (P < 0.01 for each). Multivariable Cox regression analyses, controlled for cardiovascular risk factors and comorbidities, revealed that the risk of cardiovascular death was significantly higher among RA patients with at least 3 ESR values of >or=60 mm/hour (hazard ratio [HR] 2.03, 95% confidence interval [95% CI] 1.45-2.83), RA vasculitis (HR 2.41, 95% CI 1.00-5.81), and RA lung disease (HR 2.32, 95% CI 1.11-4.84). These results indicate that markers of systemic inflammation confer a statistically significant additional risk for cardiovascular death among patients with RA, even after controlling for traditional cardiovascular risk factors and comorbidities.


The detection of microRNA associated with Alzheimer's disease in biological fluids using next-generation sequencing technologies.
Diagnostic tools for neurodegenerative diseases such as Alzheimer's disease (AD) currently involve subjective neuropsychological testing and specialized brain imaging techniques. While definitive diagnosis requires a pathological brain evaluation at autopsy, neurodegenerative changes are believed to begin years before the clinical presentation of cognitive decline. Therefore, there is an essential need for reliable biomarkers to aid in the early detection of disease in order to implement preventative strategies. microRNAs (miRNA) are small non-coding RNA species that are involved in post-transcriptional gene regulation. Expression levels of miRNAs have potential as diagnostic biomarkers as they are known to circulate and tissue specific profiles can be identified in a number of bodily fluids such as plasma, CSF and urine. Recent developments in deep sequencing technology present a viable approach to develop biomarker discovery pipelines in order to profile miRNA signatures in bodily fluids specific to neurodegenerative diseases. Here we review the potential use of miRNA deep sequencing in biomarker identification from biological fluids and its translation into clinical practice.


Reduction of plasma glycosphingolipid levels has no impact on atherosclerosis in apolipoprotein E-null mice.
Glycosphingolipids (GSLs) have been implicated as potential atherogenic lipids. Studies in apolipoprotein E-null (apoE(-/-)) mice indicate that exacerbated tissue GSL accumulation resulting from alpha-galactosidase deficiency promotes atherosclerosis, whereas the serine palmitoyl transferase inhibitor myriocin (which reduces plasma and tissue levels of several sphingolipids, including sphingomyelin, ceramide, sphingosine-1-phosphate, and GSLs) inhibits atherosclerosis. It is not clear whether GSL synthesis inhibition per se has an impact on atherosclerosis. To address this issue, apoE(-/-) mice maintained on a high-fat diet were treated with a potent glucosylceramide synthesis inhibitor, d-threo-1-ethylendioxyphenyl-2-palmitoylamino-3-pyrrolidino-propanol (EtDO-P4), 10 mg/kg/day for 94 days, and lesion development was compared in mice that were treated with vehicle only. EtDO-P4 reduced plasma GSL concentration by approximately 50% but did not affect cholesterol or triglyceride levels. Assessment of atherosclerotic lesions at four different sites indicated that EtDO-P4 had no significant impact on lesion area. Thus, despite the previously observed positive correlations between plasma and aortic GSL concentrations and the development of atherosclerosis, and the in vitro evidence implying that GSLs may be pro-atherogenic, our current data indicate that inhibition of GSL synthesis does not inhibit atherosclerosis in vivo.


Does price-cap regulation work for increasing access to contraceptives? Aggregate- and pharmacy-level evidence from Colombia.
Price caps through international reference pricing are widely used worldwide but not so commonly in over-the-counter markets. We study the short-term effects of a price cap regulation for oral contraceptives in Colombia, a market dominated by the presence of several branded generics with multiple active ingredients. Most of the regulated products were fourth-generation contraceptives, and the Colombian health benefits plan only covers second-generation ones, resulting in a de facto over-the-counter market. Our aim is to establish whether the regulation triggered a competitive response within and across product categories, by price levels and regulatory status. The panel data analysis of quarterly level data for 52 drugs (and 79 drugs in an expanded sample without transactions for some quarters) reveals a massive expansion of transactions, for the directly regulated products that were formerly the most expensive, and for the indirectly regulated (i.e., a regulated ingredient) among those with an intermediate price. Although this price reduction could have led to a crowd out of the publicly provided contraceptives, we show that this is not the case. Since the information system cannot trace the final consumers' purchases, we complement our analysis with an audit study involving 213 pharmacies in Bogota. We find that the price reduction was effectively transmitted to the final consumers.


Delay of morphine tolerance by palmitoylethanolamide.
In spite of the potency and efficacy of morphine, its clinical application for chronic persistent pain is limited by the development of tolerance to the antinociceptive effect. The cellular and molecular mechanisms underlying morphine tolerance are complex and still unclear. Recently, the activation of glial cells and the release of glia-derived proinflammatory mediators have been suggested to play a role in the phenomenon. N-palmitoylethanolamine (PEA) is an endogenous compound with antinociceptive effects able to reduce the glial activation. On this basis, 30 mg kg(-1) PEA was subcutaneously daily administered in morphine treated rats (10 mg kg(-1) intraperitoneally, daily). PEA treatment significantly attenuated the development of tolerance doubling the number of days of morphine antinociceptive efficacy in comparison to the vehicle + morphine group. PEA prevented both microglia and astrocyte cell number increase induced by morphine in the dorsal horn; on the contrary, the morphine-dependent increase of spinal TNF-α levels was not modified by PEA. Nevertheless, the immunohistochemical analysis revealed significantly higher TNF-α immunoreactivity in astrocytes of PEA-protected rats suggesting a PEA-mediated decrease of cytokine release from astrocyte. PEA intervenes in the nervous alterations that lead to the lack of morphine antinociceptive effects; a possible application of this endogenous compound in opioid-based therapies is suggested.


Stress induction of the mammalian GRP78/BiP protein gene: in vivo genomic footprinting and identification of p70CORE from human nuclear extract as a DNA-binding component specific to the stress regulatory element.
GRP78, also known as BiP, is one of the better-characterized molecular chaperones. It has been implicated in protein folding and also calcium sequestration in the endoplasmic reticulum. When the cells are subjected to endoplasmic reticulum stress, in particular the depletion of stored calcium and/or the accumulation of abnormal proteins, the rate of transcription of grp78 is enhanced. Previous studies have shown that the core region of the rat grp78 promoter (-170 to -135), which is 95% conserved with the human grp78 core (-133 to -98), is one of the key regulatory elements. Using ligation-mediated PCR, we have found that there are specific changes in factor occupancy after stress induction and the major changes occur within a cluster of bases located in the 3' half of the grp core, whereas other regulatory elements are constitutively occupied. This inducible binding to the 3' half of the human grp78 core region is observed under diverse stress signals, suggesting a common mechanism for the grp stress response. Nonetheless, the lack of constitutive in vivo protection at this region is not due to the absence of a binding factor in nuclear extracts. Using in vitro gel mobility shift assays, we detected a constitutive binding activity which exhibits specificity and affinity to the stress-inducible region. Through sodium dodecyl sulfate-polyacrylamide gel electrophoresis size fractionation and renaturation analysis, the activity is found in polypeptides with molecular sizes of 65 to 75 kDa. After a three-step purification scheme including core affinity column chromatography, we purified p70CORE, which is about 70 kDa in its monomeric form. The purified p70CORE is sufficient to form a complex specific to the stress-inducible region.


Pathways to psychiatric care in Ibadan, Nigeria.
Pathways patients take to psychiatric care will reflect the nature of the services available and the popular beliefs about mental illness. Studying the pathways may help in the identification of sources of delay in the receipt of care and suggest possible improvements. By studying the pathways of 159 patients to a tertiary psychiatric service in Nigeria, we show that traditional and religious healers are consulted at some stage by many patients with mental illness. Such healers are the first carers to a large proportion of the patients. These patients are not different from those who consult orthodox medical practitioners either in demographic features, presenting complaints, or nearness to service. Patients who consult traditional healers first tended to arrive at a tertiary psychiatric service much later than those who consult other carers. Our observations suggest that attempts to incorporate traditional medical care into the health care system must seek to improve their referral skill.