Neoplastic transformation of C3H/10T1/2 cells following exposure to 120-Hz modulated 2.45-GHz microwaves and phorbol ester tumor promoter.
Some recent epidemiological studies have shown a positive association between cancer incidence and exposure to electromagnetic (EM) fields. Evidence from in vitro studies indicates that this effect could be due to synergistic interaction between EM fields and tumor promoters. However, no dose-response data related directly to carcinogenesis have been published. In this study, actively growing cultures of C3H/10T1/2 cells were exposed for 24 h to 2.45-GHz microwaves pulse-modulated at 120 Hz. Conditions of EM-field exposure were designed to simulate low-field exposures (specific absorption rate 0.1, 1, or 4.4 W/kg; the corresponding peak amplitudes were electric field 18, 56, or 120 V/m, magnetic field 0.09, 0.27, or 0.56 muT, respectively). In separate experiments, a 24-h EM-field exposure at 4.4 W/kg was preceded or followed by X irradiation at 0.5, 1, or 1.5 Gy. Cells were assayed for cell survival and neoplastic transformation with or without post-treatment administration of 0.1 micrograms/ml of 12-O-tetradecanoylphorbol-13-acetate (TPA) for the duration of the assay. The EM fields alone had no effect on cell survival or induction of neoplastic transformation. However, enhancement of transformation due to EM fields plus TPA was highly significant and ranged up to a level equivalent to that produced by 1.5 Gy of X rays. The frequency of neoplastic transformation was dependent on the level of EM exposure and was additive with doses of X rays given as a cocarcinogen.


A validation study comparing accelerator MS and liquid scintillation counting for analysis of 14C-labelled drugs in plasma, urine and faecal extracts.
A comparison has been made between accelerator mass spectrometry (AMS) analysis and liquid scintillation counting (LSC) of plasma, urine and faecal samples containing 14C-labelled drugs. In an in vitro study in which human plasma was spiked (the term spiked is used in Section 2.6) with 14C-Fluconazole (14C-FL) over a concentration range of 0.1-2.5 dpm/ml, a correlation coefficient of 0.999 was determined for AMS analysis versus extrapolated LSC data. No significant day to day (or inter-day)variation was seen (P < 0.05 by ANOVA). Coefficients of variation for these analyses ranged from 2.68 to 6.50%. In vivo studies in which rats were given a high (11.5 microCi/kg) or low (18.1 nCi/kg) radioactive dose (to model an exposure of 0.9 microSievert to man) of 14C-Fluticasone propionate(14C-FP) showed that there was also a good correspondence between AMS and LSC data. A mass balance study in a single the faeces by 96 h; less than 1% of the administered dose was excreted in the urine. The limit of reliable measurement of drug related material, above background concentrations, by AMS analysis in this study was approximately 0.1 dpm/ml for plasma, 0.01 dpm/ml for urine without any sample extraction or concentration and 0.01 dpm/ml for faecal extracts. The data reported here demonstrate that AMS is an ultrasensitive and reliable method for analysing 14C-labelled drugs in human and animal body fluids.


Human homolog of patched, a candidate gene for the basal cell nevus syndrome.
The basal cell nevus syndrome (BCNS) is characterized by developmental abnormalities and by the postnatal occurrence of cancers, especially basal cell carcinomas (BCCs), the most common human cancer. Heritable mutations in BCNS patients and a somatic mutation in a sporadic BCC were identified in a human homolog of the Drosophila patched (ptc) gene. The ptc gene encodes a transmembrane protein that in Drosophila acts in opposition to the Hedgehog signaling protein, controlling cell fates, patterning, and growth in numerous tissues. The human PTC gene appears to be crucial for proper embryonic development and for tumor suppression.


Cannabidiol causes activated hepatic stellate cell death through a mechanism of endoplasmic reticulum stress-induced apoptosis.
The major cellular event in the development and progression of liver fibrosis is the activation of hepatic stellate cells (HSCs). Activated HSCs proliferate and produce excess collagen, leading to accumulation of scar matrix and fibrotic liver. As such, the induction of activated HSC death has been proposed as a means to achieve resolution of liver fibrosis. Here we demonstrate that cannabidiol (CBD), a major non-psychoactive component of the plant Cannabis sativa, induces apoptosis in activated HSCs through a cannabinoid receptor-independent mechanism. CBD elicits an endoplasmic reticulum (ER) stress response, characterized by changes in ER morphology and the initiation of RNA-dependent protein kinase-like ER kinase-, activating transcription factor-6-, and inositol-requiring ER-to-nucleus signal kinase-1 (IRE1)-mediated signaling cascades. Furthermore, CBD induces downstream activation of the pro-apoptotic IRE1/ASK1/c-Jun N-terminal kinase pathway, leading to HSC death. Importantly, we show that this mechanism of CBD-induced ER stress-mediated apoptosis is specific to activated HSCs, as it occurs in activated human and rat HSC lines, and in primary in vivo-activated mouse HSCs, but not in quiescent HSCs or primary hepatocytes from rat. Finally, we provide evidence that the elevated basal level of ER stress in activated HSCs has a role in their susceptibility to the pro-apoptotic effect of CBD. We propose that CBD, by selectively inducing death of activated HSCs, represents a potential therapeutic agent for the treatment of liver fibrosis.


Red light interferes in UVA-induced photoaging of human skin fibroblast cells.
The possible regulation mechanism of red light was determined to discover how to retard UVA-induced skin photoaging. Human skin fibroblasts were cultured and irradiated with different doses of UVA, thus creating a photoaging model. Fibroblasts were also exposed to a subtoxic dose of UVA combined with a red light-emitting diode (LED) for five continuous days. Three groups were examined: control, UVA and UVA plus red light. Cumulative exposure doses of UVA were 25 J cm(-2), and the total doses of red light were 0.18 J cm(-2). Various indicators were measured before and after irradiation, including cell morphology, viability, β-galactosidase staining, apoptosis, cycle phase, the length of telomeres and the protein levels of photoaging-related genes. Red light irradiation retarded the cumulative low-dose UVA irradiation-induced skin photoaging, decreased the expression of senescence-associated β-galactosidase, upregulated SIRT1 expression, decreased matrix metalloproteinase MMP-1 and the acetylation of p53 expression, reduced the horizon of cell apoptosis and enhanced cell viability. Furthermore, the telomeres in UVA-treated cells were shortened compared to those of cells in the red light groups. These results suggest that red light plays a key role in the antiphotoaging of human skin fibroblasts by acting on different signaling transduction pathways.


A hybrid method for quantifying China's nitrogen footprint during urbanisation from 1990 to 2009.
In this study, we devise a national nitrogen footprint method to evaluate the life cycle nitrogen flows through the national economy of China from 1990 to 2009. To this end, we build a hybrid method based on two well-established techniques, namely material flow analysis (MFA) and input-output analysis (IOA). This integration allows for the evaluation of the effects of international trade and interdependencies among economic sectors. Our results suggest that China's nitrogen footprint (NF) has increased from 30.3Teragrams (Tg) in 1990 to 54.0Tg in 2009, whereas the NF per capita has increased from 25.9 to 39.5kgN/yr. Relationship between the world NF per capita and human development index (HDI) appears to show an inverted U curve, whilst China shows an increase both in NF per capita and HDI. We find that an increase in China's NF is largely associated with high levels of urbanisation. Although the energy NF (E_NF) has increased more drastically than the food NF (F_NF), the latter still dominates China's total NNF, with proportions of 91% in 1990 and 82% in 2009. Taking international trade into account, our results demonstrate that China was a net exporter of F_NF, whilst a net importer of E_NF over this time period. There are many measures considered to reduce China's nitrogen footprint, including improvements in N use efficiency of food systems, transformation of meat-based diets and optimisation of China's economic structure.


Assessment of microalbuminuria for early diagnosis and risk prediction in dengue infections.
Dengue is the most important arboviral infection of humans. Following an initial febrile period, a small proportion of infected patients develop a vasculopathy, with children at particular risk for severe vascular leakage and shock. Differentiation between dengue and other common childhood illnesses is difficult during the early febrile phase, and risk prediction for development of shock is poor. The presence of microalbuminuria is recognized as a useful early predictor for subsequent complications in a number of other disorders with vascular involvement. Significant proteinuria occurs in association with dengue shock syndrome and it is possible that early-phase microalbuminuria may be helpful both for diagnosis of dengue and for identification of patients likely to develop severe disease. We measured formal urine albumin to creatinine ratios (UACRs) in daily samples obtained from a large cohort of children with suspected dengue recruited at two outpatient clinics in Ho Chi Minh City, Vietnam. Although UACRs were increased in the 465 confirmed dengue patients, with a significant time trend showing peak values around the critical period for dengue-associated plasma leakage, urine albumin excretion was also increased in the comparison group of 391 patients with other febrile illnesses (OFI). The dengue patients generally had higher UACRs than the OFI patients, but microalbuminuria, using the conventional cutoff of 30 mg albumin/g creatinine discriminated poorly between the two diagnostic groups in the early febrile phase. Secondly UACRs did not prove useful in predicting either development of warning signs for severe dengue or need for hospitalization. Low-level albuminuria is common, even in relatively mild dengue infections, but is also present in many OFIs. Simple point-of-care UACR tests are unlikely to be useful for early diagnosis or risk prediction in dengue endemic areas.


The effect of low-level normobaric hypoxia on orthostatic responses.
Altitude-induced hypoxia is believed to decrease tolerance to orthostasis. This preliminary investigation assesses the interaction between low-level normobaric hypoxia and the cardiovascular responses to orthostasis. We hypothesize that low-level hypoxia at simulated altitudes up to 3350 m adversely affects cardiovascular responses to orthostasis. Six women and eight men underwent + 75 degrees head-up tilt (HUT) at 3 simulated altitudes (115 m (377 ft), 1680 m (5500 ft) and 3350 m (11,000 ft)). Minute ventilation (VE), tidal volume (VT), breathing frequency (f), heart rate (HR), and arterial blood pressure (BP) were measured non-invasively. Ventilatory parameters (VE, VT, and f) were measured during the control period to assess the respiratory response to hypoxia. Cardiovascular variables (HR and BP) during HUT were compared and analyzed by calculating the deviation from control (pre-tilt) values. Respiratory data showed no significant hypoxic ventilatory response. The average HR deviation significantly decreased (p < 0.05) with HUT from 13 +/- 2 bpm (115 m) to 9 +/- 2 bpm (3350 m). The pattern of mean arterial BP deviation with HUT was similar across the three altitudes (-6.6 +/- 2.9 mm Hg at 115 m to -6.8 +/- 2.4 mm Hg at 3350 m), with no significant differences. Changes in HR and arterial BP are fundamental aspects of the cardiovascular response to orthostatic stress. Although there were no significant changes in the BP response to orthostasis with hypoxia, the HR changes seen in this study suggest that the ability to modulate HR under orthostatic stress is reduced with exposure to low-level normobaric hypoxia.


Frequency, cost and impact of inter-island referrals in the Solomon Islands.
Providing quality health services to people living in remote areas is central to global efforts to achieve universal access to health care. Effective referral systems are especially critical in resource-limited countries where small populations are separated by considerable distances, geographic challenges and the limitations of human resources for health. This study aimed to build an evidence base on inter-island referrals in the Solomon Islands, in particular regarding the number of referrals, reasons for referrals, and cost, to ultimately provide recommendations regarding referral practice effectiveness and efficiency. Data were taken from the referral database collected and maintained by the National Referral Hospital (NRH) in the capital, Honiara. Data included age, sex, ward or department visited, date of travel back to home port, home port and province. Data were available and included for 2008, 6 months of 2009, all of 2010 and 1 month of 2011; a total of 31 months. Travel costs were taken from NRH administrative information and included in the analysis. In addition, 10 qualitative interviews were conducted with clinicians and policy-makers in the tertiary hospital and one provincial hospital to gather information regarding inter-island referrals, their appropriateness and challenges faced. In the Solomon Islands, referrals from outer islands to the NRH are substantial and are gradually increasing over time. The two most populous provinces outside of the capital, Western and Malaita, represented 51% of all referrals in the study period. Of those referred, 21% were less than 15 years of age - even though 40% of the country's population is under 15 - with 30% being young adults of 15-24 years. Orthopaedic conditions comprised the largest number of referrals, with obstetric and gynaecological conditions a close second. The cost of referrals is rapidly increasing and was almost US$350,000 per year for the NRH alone. The amount budgeted for patient travel from the provinces to the NRH was a fraction of what is needed to cover the current number of referrals leading to a substantial budget shortfall. There did not appear to be a clear link between number of doctors in each province and the rate of referrals. Improving the appropriateness of referrals can have a substantial impact on access, quality of care and costs. Improvements in equipment in remote facilities, in human resources for health and in information technology can strengthen the quality of care in outer islands. Reducing the burden on referral facilities will allow them to provide appropriate care to those most in need while building public trust in all layers of the health system.


Evidence for a vascular contribution to diffusion FMRI at high b value.
Recent work has suggested that diffusion-weighted functional magnetic resonance imaging (FMRI) with strong diffusion weighting (high b value) detects neuronal swelling that is directly related to neuronal firing. This would constitute a much more direct measure of brain activity than current methods and represent a major advance in neuroimaging. However, it has not been firmly established that the observed signal changes do not reflect residual vascular effects, which are known to exist at low b value. This study measures the vascular component of diffusion FMRI directly by using hypercapnia, which induces blood flow changes in the absence of a change in neuronal firing. Hypercapnia elicits a similar diffusion FMRI response to a visual stimulus including a rise in percent signal change with increasing b value, which was reported for visual activation. Analysis of the response timing found no evidence for an early response at high b value, which has been reported as evidence for a nonhemodynamic response. These results suggest that a large component of the diffusion FMRI signal at high b value is vascular rather than neuronal.


Primary care and public health services integration in Brazil's unified health system.
We examined associations between transdisciplinary collaboration, evidence-based practice, and primary care and public health services integration in Brazil's Family Health Strategy. We aimed to identify practices that facilitate service integration and evidence-based practice. We collected cross-sectional data from community health workers, nurses, and physicians (n = 262). We used structural equation modeling to assess providers' service integration and evidence-based practice engagement operationalized as latent factors. Predictors included endorsement of team meetings, access to and consultations with colleagues, familiarity with community, and previous research experience. Providers' familiarity with community and team meetings positively influenced evidence-based practice engagement and service integration. More experienced providers reported more integration and engagement. Physicians reported less integration than did community health workers. Black providers reported less evidence-based practice engagement than did Pardo (mixed races) providers. After accounting for all variables, evidence-based practice engagement and service integration were moderately correlated. Age and race of providers, transdisciplinary collaboration, and familiarity with the community are significant variables that should inform design and implementation of provider training. Promising practices that facilitate service integration in Brazil may be used in other countries.


Associations between the Home Environment, Feeding Practices and Children's Intakes of Fruit, Vegetables and Confectionary/Sugar-Sweetened Beverages.
Within the home environment, parents influence their children's dietary intakes through their parenting and dietary practices, and the foods they make available/accessible. The aim of this cross-sectional study was to examine the associations between home environmental characteristics and children's dietary intakes. Three hundred and thirty-two children aged three-five years and their parents participated in the study. Home environmental characteristics, including parental control feeding practices, were explored using validated and standardized questionnaires such as the Child Feeding Questionnaire (CFQ), the Physical and Nutritional Home Environment Inventory (PNHEI) and the Healthy Home Survey (HHS). Parent and child food consumption was also measured. Pressure to eat from parents was associated with lower fruit intake in children (OR 0.67, 95% CI 0.47-0.96, p = 0.032). Greater variety of fruit available in the home increased the likelihood of fruit consumption in children (OR 1.35 95% CI 1.09-1.68, p = 0.005). Watching television for ≥1 h per day was associated with a decreased probability of children eating vegetables daily (OR 0.38, 95% CI 0.20-0.72, p = 0.003) and doubled their likelihood of consuming confectionary/sugar-sweetened beverages more than once weekly (OR 2.15, 95% CI 1.06-4.38, p = 0.034). Children whose parents had lower vegetable consumption were 59% less likely to eat vegetables daily. This study demonstrates that modifiable home environmental characteristics are significantly associated with children's dietary intakes.


Resolution of Laryngeal Oedema in a Patient with Acquired C1-Inhibitor Deficiency. a Case Report.
Laryngeal oedema caused by acquired angioedema due to C1-inhibitor deficiency (C1-INH-AAE) is a life-threatening condition. The swelling is bradykinin mediated and will not respond to the usual treatment with antihistamines, corticosteroids, or epinephrine. Instead, kallikrein-bradykinin-targeted therapies should be used promptly to prevent asphyxiation. A 43 years old female presented at the Hereditary Angioedema Centre reporting a one-year history of peripheral, facial, and neck oedema. Treatment with antihistamines and corticosteroids had been ineffective. Laboratory results showed complement level deficiencies and monoclonal gammopathy characterised as immunoglobulin M. An abdominal ultrasound revealed splenomegaly. A bone marrow biopsy was normal. Based on these data, the diagnosis of C1-INH-AAE associated with monoclonal gammopathy of uncertain significance (MGUS) was made. As C1-INH-AAE can present with life-threatening, standard treatment-resistant laryngeal oedema, an emergency care treatment plan was proposed, and the patient was advised to present to the emergency department (ED) with this medical letter. Based on these recommendations, three laryngeal attacks were successfully treated in the ED with recombinant human C1-inhibitor (two attacks) and fresh frozen plasma (one attack). After these episodes, the patient was prescribed prophylactic treatment with antifibrinolytics. No further angioedema attacks were reported by the patient at the 18 months follow-up visit. Because angioedema of the upper airways is a life-threatening condition, recognising the specific type of swelling by the emergency physician is critical in providing immediate and effective treatment to reduce the associated risk of asphyxiation. C1-INH-AAE being a rare disorder, patients should have available an emergency care treatment plan with recommendations of acute treatment possibilities.


Plasma Glial Fibrillary Acidic Protein Levels Differ Along the Spectra of Amyloid Burden and Clinical Disease Stage.
Measuring plasma glial fibrillary acidic protein (GFAP) alongside cortical amyloid-β (Aβ) may shed light on astrocytic changes in aging and Alzheimer's disease (AD). To examine associations between plasma GFAP and cortical Aβ deposition in older adults across the typical aging-to-AD dementia spectrum. We studied two independent samples from UCSF (Cohort 1, N = 50; Cohort 2, N = 37) covering the spectra of clinical severity (CDR Sum of Boxes; CDR-SB) and Aβ-PET burden. Aβ-PET was completed with either florbetapir or Pittsburgh Compound B and standardized uptake value ratios were converted to the Centiloid (CL) scale for analyses. All participants with CDR-SB > 0 were Aβ-PET positive, while clinically normal participants (CDR-SB = 0) were a mix of Aβ-PET positive and negative. Regression analyses evaluated main effect and interaction associations between plasma GFAP, Aβ-PET, and clinical severity. In both cohorts, plasma GFAP increased linearly with Aβ-PET CLs in clinically normal older adults. In Cohort 2, which included participants with more severe clinical dysfunction and Aβ-PET burden, the association between Aβ and GFAP became curvilinear (inverted U-shape; quadratic model R2 change = 0.165, p = 0.009), and Aβ-PET interacted with CDR-SB (R2 change = 0.164, p = 0.007): older adults with intermediate functional impairment (CDR-SB = 0.5-4.0) showed a weak (negative) association between Aβ-PET CLs and plasma GFAP, while older adults with dementia (CDR-SB > 4.0) showed a strong, negative association of higher Aβ-PET CLs with lower plasma GFAP. The relationship between astrocytic integrity and cortical Aβ may be highly dynamic, with linear, positive associations early in disease that diverge in more severe disease stages.


A randomised controlled trial of cognitive-behaviour therapy for clinical perfectionism: a preliminary study.
Perfectionism can be a problem in its own right and it can impede the progress of treatment of Axis I disorders. This study reports on a preliminary randomised controlled trial of cognitive-behaviour therapy (CBT) for "clinical perfectionism". Twenty participants were randomly assigned to either immediate treatment (IT) (n=10) or a wait list (NL) (n=10). Treatment consisted of ten sessions of CBT over eight weeks. Two participants did not complete the follow-up assessments (10%). Fifteen of the original 20 participants (75%) were clinically significantly improved after treatment and the effect size was large (1.8). Treatment gains were maintained at 8-week and 16-week follow-up.


Evaluation of an On-Farm Culture System (Accumast) for Fast Identification of Milk Pathogens Associated with Clinical Mastitis in Dairy Cows.
The present study aimed evaluate an on-farm culture system for identification of milk pathogens associated with clinical mastitis in dairy cows using two different gold standard approaches: standard laboratory culture in study 1 and 16S rRNA sequencing in study 2. In study 1, milk from mastitic quarters (i.e. presence of flakes, clots, or serous milk; n = 538) was cultured on-farm using a single plate containing three selective chromogenic media (Accumast-FERA Animal Health LCC, Ithaca, NY) and in a reference laboratory using standard culture methods, which was considered the gold standard. In study 2, mastitic milk was cultured on-farm and analyzed through 16S rRNA sequencing (n = 214). In both studies, plates were cultured aerobically at 37°C for 24 h and read by a single technician masked to gold standard results. Accuracy, sensitivity, specificity, positive (PPV) and negative predictive value (NPV) were calculated based on standard laboratory culture in study 1, and PPV was calculated based on sequencing results in study 2. Overall accuracy of Accumast was 84.9%. Likewise, accuracy for identification of Gram-negative bacteria, Staphylococcus sp., and Streptococcus sp. was 96.4%, 93.8%, and 91.5%, respectively. Sensitivity, specificity, PPV, and NPV were 75.0%, 97.9%, 79.6%, and 97.3% for identification of E. coli, 100.0%, 99.8%, 87.5%, and 100.0% for S. aureus, 70.0%, 95.0%, 45.7%, and 98.1% for other Staphylococcus sp., and 90.0%, 92.9%, 91.8%, and 91.2% for Streptococcus sp. In study 2, Accumast PPV was 96.7% for E. coli, 100.0% for Enterococcus sp., 100.0% for Other Gram-negatives, 88.2% for Staphylococcus sp., and 95.0% for Streptococcus sp., respectively. In conclusion, Accumast is a unique approach for on-farm identification pathogens associated with mastitis, presenting overall sensitivity and specificity of 82.3% and 89.9% respectively.


Care arrangements for community-dwelling people with dementia in Germany as perceived by informal carers - a cross-sectional pilot survey in a provincial-rural setting.
The majority of people with dementia live at home, and informal carers assume the role of key care providers, often supported by formal services. The purpose of this pilot study was to assess home-based care arrangements, to illustrate utilisation of formal services over time and to identify factors associated with perceived stability of the care situation from the informal carer's perspective. A self-administered questionnaire (D-IVA 'Instrument for Assessing Home-Based Care Arrangements for People with Dementia') was developed and distributed in a provincial-rural setting in Germany as a cross-sectional survey. Data analysis used descriptive statistics, unbiased conditional inference trees and thematic analysis for open-ended questions. In total, 84 care arrangements were assessed. The majority of participants were direct relatives of the care-dependent person [mostly adult children (48.8%) or spouses (27.4%)]. Formal services were already sought in the first year after onset of memory problems. The most frequently used formal services were home care nursing services (53.0%), day care (49.4%) and respite care (29.6%), whereas 15.5% did not use any type of formal support. Companion home visit, home care nursing service and day care were used over the longest periods of time. The recruitment strategy used in this study may have recruited persons who were relatively more dependent on their informal carers. In this small sample, carers' perceived stability of the care situation was high, and this was associated with the country of origin and sex of the person with dementia (P = 0.004 and 0.023 respectively). Most care arrangements consisted of a mix of informal and formal services. However, informal carers assumed prime responsibility. The questionnaire D-IVA proved to be suitable. It remains a challenge to further examine factors associated with perceived stability and to explain the phenomenon in its whole complexity. Further research using the D-IVA should consider applying complementing quantitative measures as well as qualitative methods.


Repeat kidney biopsies fail to detect differences between azathioprine and mycophenolate mofetil maintenance therapy for lupus nephritis: data from the MAINTAIN Nephritis Trial.
In the MAINTAIN Nephritis Trial, azathioprine (AZA) and mycophenolate mofetil (MMF) were compared as maintenance immunosuppressive treatment of proliferative lupus nephritis (LN) after a short-course of intravenous cyclophosphamide. Here, we compare the pathological findings on repeat kidney biopsies between the two groups. Per protocol, repeat renal biopsies were obtained in 30 patients (16 AZA and 14 MMF) at 2 years (±6 months). Baseline and follow-up biopsies were graded according to the International Society of Nephrology/Renal Pathological Society (ISN/RPS) classification. The activity and chronicity indices (AI, CI) were calculated using two different semiquantitative scoring systems (Morel-Maroger and National Institutes of Health). Statistics were performed by non-parametric tests. The clinical characteristics of the 30 re-biopsied patients only marginally differ from the entire MAINTAIN cohort (105 patients). Clinical baseline and follow-up characteristics of AZA- and MMF-treated re-biopsied patients did not differ. Time (SD) to repeat renal biopsy was 25.0 (2.0) and 26.5 (3.3) months in AZA and MMF patients, respectively. More patients had normal renal biopsies or Classes I/II/V LN at follow-up compared to baseline and conversely, less patients had Class IV LN at follow-up. In both groups, the AI statistically decreased at follow-up compared to baseline, while the CI slightly, but significantly, increased. No differences could be detected between the groups. Centralized pathological analyses, including ISN/RPS classification and comparisons of AI/CI, failed to find differences between MMF and AZA at 2 years, a result well in line with the absence of difference in long-term clinical outcome reported elsewhere.


A claims-based view of health care charges and utilization for commercially insured patients with osteoarthritis.
To evaluate managed care medical and pharmacy claims data from commercially insured patients with osteoarthritis and to examine cost and resource utilization patterns across inpatient care, outpatient care, emergency department, and pharmaceuticals. Retrospective claims-based analysis. Data were obtained from the PharMetrics/IMS Integrated Patient-Centric Database which contains medical and pharmacy claims for over 60 million Americans. Patients were selected by the presence of one or more Episode Treatment Groups (721* and 722*) associated with joint degeneration and then further segmented by ICD-9 codes (715*) to create a homogeneous group of patients with diagnosed osteoarthritis. Patients were then further stratified into groups for evaluation based on demographics and clinical variables and their effect on cost and utilization patterns. Patients were eligible for inclusion if they fulfilled the ETG and ICD-9 code criteria from Jan. 1, 2007 through Dec. 31, 2007 and possessed 12 months of plan eligibility. Over 1.1 million patients were included in the analysis. The average age was 54 years, and over 68% of the patients were between ages 36 and 64. Average annual charges associated with osteoarthritis treatment were $5,938, with about 40% from the inpatient environment, 53% from outpatient care, and only 6.3% from pharmaceuticals. The most commonly utilized pharmacotherapies were narcotic analgesics, NSAIDs, and corticosteroids. There was also significant utilization of prescription proton-pump inhibitors and H2 antagonists. These osteoarthritis-specific, population-based measurements are an exercise in developing data that are easily reproducible in a managed care environment. These data offer a starting point and comparator to further investigate and contrast the charges associated with osteoarthritis and the utilization of ancillary pharmacotherapeutic agents.


Maternal occupational exposures and risk of spontaneous abortion in veterinary practice.
To examine the relationship between occupational exposures and spontaneous abortion in female veterinarians. The Health Risks of Australian Veterinarians project (HRAV) was a questionnaire-based survey of all graduates from Australian veterinary schools from 1960 to 2000. Of 5748 eligible veterinarians sent the questionnaires, 2800 replied including 1197 females (42.8%). The response rate was 59% of women veterinarians eligible to participate. The pregnancy of women was defined as the unit of analysis. We restricted analyses to pregnancies of those women who reported being employed when the pregnancy began and were working only in clinical practice. Of 1355 pregnancies, 940 were eligible for the final analysis. Self-reported occupational exposures to anaesthetic gases, x rays, pesticides and long working hours in relation to spontaneous abortion were examined. In a multiple logistic regression controlling for 12 potential confounders, there was a more than twofold significant increase (OR 2.49, 95% CI 1.02 to 6.04) in the risk of spontaneous abortion in women exposed to unscavenged anaesthetic gases for > or =1 h per week. Veterinarians who reported performing more than five radiographic examinations per week had a statistically significant elevated risk of spontaneous abortion compared to those who performed five or less (OR 1.82, 95% CI 1.17 to 2.82). There was also approximately a twofold significant increased risk of spontaneous abortion in women who used pesticides at work (OR 1.88, 95% CI 1.18 to 3.00). Female veterinarians, particularly those of childbearing age, should be fully informed of the possible reproductive effects of unscavenged anaesthetic gases, ionising radiation and pesticide exposure and reduce their exposure by using protective devices when they are planning to become pregnant and during pregnancy.


Creating new evolutionary pathways through bioinvasion: the population genetics of brushtail possums in New Zealand.
Rapid increases in global trade and human movement have created novel mixtures of organisms bringing with them the potential to rapidly accelerate the evolution of new forms. The common brushtail possum (Trichosurus vulpecula), introduced into New Zealand from Australia in the 19th century, is one such species having been sourced from multiple populations in its native range. Here, we combine microsatellite DNA- and GIS-based spatial data to show that T. vulpecula originating from at least two different Australian locations exhibit a population structure that is commensurate with their introduction history and which cannot be explained by landscape features alone. Most importantly, we identify a hybrid zone between the two subspecies which appears to function as a barrier to dispersal. When combined with previous genetic, morphological and captive studies, our data suggest that assortative mating between the two subspecies may operate at a behavioural or species recognition level rather than through fertilization, genetic incompatibility or developmental inhibition. Nevertheless, hybridization between the two subspecies of possum clearly occurs, creating the opportunity for novel genetic combinations that would not occur in their natural ranges and which is especially likely given that multiple contact zones occur in New Zealand. This discovery has implications for wildlife management in New Zealand because multiple contact zones are likely to influence the dispersal patterns of possums and because differential susceptibility to baiting with sodium fluoroacetate between possums of different origins may promote novel genetic forms.


Parenteral anticoagulants: American College of Chest Physicians Evidence-Based Clinical Practice Guidelines (8th Edition).
This chapter describes the pharmacology of approved parenteral anticoagulants, including the indirect anticoagulants, unfractionated heparin (UFH), low-molecular-weight heparins (LMWHs), fondaparinux, and danaparoid as well as the direct thrombin inhibitors hirudin, bivalirudin, and argatroban. UFH is a heterogeneous mixture of glycosaminoglycans that bind to antithrombin via a unique pentasaccharide sequence and catalyze the inactivation of thrombin factor Xa and other clotting factors. Heparin also binds to cells and other plasma proteins, endowing it with unpredictable pharmacokinetic and pharmacodynamic properties, and can lead to nonhemorrhagic side effects, such as heparin-induced thrombocytopenia (HIT) and osteoporosis. LMWHs have greater inhibitory activity against factor Xa than thrombin and exhibit less binding to cells and proteins than heparin. Consequently, LMWH preparations have more predictable pharmacokinetic and pharmacodynamic properties, have a longer half-life than heparin, and have a lower risk of nonhemorrhagic side effects. LMWHs can be administered once or twice daily by subcutaneous injection, without anticoagulant monitoring. Based on their greater convenience, LMWHs have replaced UFH for many clinical indications. Fondaparinux, a synthetic pentasaccharide, catalyzes the inhibition of factor Xa, but not thrombin, in an antithrombin-dependent fashion. Fondaparinux binds only to antithrombin; therefore, HIT and osteoporosis are unlikely to occur. Fondaparinux has excellent bioavailability when administered subcutaneously, has a longer half-life than LMWHs, and is given once daily by subcutaneous injection in fixed doses, without anticoagulant monitoring. Three parenteral direct thrombin inhibitors and danaparoid are approved as alternatives to heparin in HIT patients.


OLAND is feasible to treat sewage-like nitrogen concentrations at low hydraulic residence times.
Energy-positive sewage treatment can, in principle, be obtained by maximizing energy recovery from concentrated organics and by minimizing energy consumption for concentration and residual nitrogen removal in the main stream. To test the feasibility of the latter, sewage-like nitrogen influent concentrations were treated with oxygen-limited autotrophic nitrification/denitrification (OLAND) in a lab-scale rotating biological contactor at 25°C. At influent ammonium concentrations of 66 and 29 mg N L(-1) and a volumetric loading rate of 840 mg N L(-1) day(-1) yielding hydraulic residence times (HRT) of 2.0 and 1.0 h, respectively, relatively high nitrogen removal rates of 444 and 383 mg N L(-1) day(-1) were obtained, respectively. At low nitrogen levels, adapted nitritation and anammox communities were established. The decrease in nitrogen removal was due to decreased anammox and increased nitratation, with Nitrospira representing 6% of the biofilm. The latter likely occurred given the absence of dissolved oxygen (DO) control, since decreasing the DO concentration from 1.4 to 1.2 mg O(2) L(-1) decreased nitratation by 35% and increased anammox by 32%. Provided a sufficient suppression of nitratation, this study showed the feasibility of OLAND to treat low nitrogen levels at low HRT, a prerequisite to energy-positive sewage treatment.


Transforming an industrial giant. Interview by Thomas A. Stewart and Louise O'Brien.
In his 12 years at the helm of Siemens, CEO Heinrich von Pierer designed and directed a major transformation. Taking this German icon from a technically superb but slow-moving industrial giantto a disciplined yet nimble multinational has posed enormous challenges. Since 1992, Siemens has revamped its portfolio of businesses, expanded its reach into 192 countries, and created a more local-market-driven culture, gaining recognition as one of the best-managed and most competitive companies in the world. In this edited interview with HBR editor Thomas A. Stewart and consulting editor Louise O'Brien, von Pierer describes the requirements for transformation and culture change and how he broke down historical barriers at Siemens. He shares his insights about portfolio restructuring, his lessons from competing with GE, and the pros and cons of being based in Europe versus America. He reflects on the true start of globalization after the fall of the Berlin wall and on how dramatically the company needed to change in order to counter the resulting pricing pressures across all of its businesses. He talks, too, about the biggest challenge on his successor's desk-"the particular challenge of China;" he says. Amid all these topics, von Pierer reiterates the importance of people: "We all talk about people as our most important resource, but as a matter of fact, who's really taking care of people?... We need [their] backing. We can't afford to run into a situation where people no longer accept what we do."


Speech telepractice: installing a speech therapy upgrade for the 21st century.
Much of speech therapy involves the clinician guiding the therapeutic process (e.g., presenting stimuli and eliciting client responses). However, this Brief Communication describes a different approach to speech therapy delivery. Clinicians at Waldo County General Hospital (WCGH) use high definition audio and video to engage clients in telepractice using interactive web-based virtual environments. This technology enables clients and their clinicians to co-create salient treatment activities using authentic materials captured via digital cameras, video and/or curricular materials. Both therapists and clients manipulate the materials and interact online in real-time. The web-based technology engenders highly personalized and engaging activities, such that clients' interactions with these high interest tasks often continue well beyond the therapy sessions.


Amyloid-dependent triosephosphate isomerase nitrotyrosination induces glycation and tau fibrillation.
Alzheimer's disease neuropathology is characterized by neuronal death, amyloid beta-peptide deposits and neurofibrillary tangles composed of paired helical filaments of tau protein. Although crucial for our understanding of the pathogenesis of Alzheimer's disease, the molecular mechanisms linking amyloid beta-peptide and paired helical filaments remain unknown. Here, we show that amyloid beta-peptide-induced nitro-oxidative damage promotes the nitrotyrosination of the glycolytic enzyme triosephosphate isomerase in human neuroblastoma cells. Consequently, nitro-triosephosphate isomerase was found to be present in brain slides from double transgenic mice overexpressing human amyloid precursor protein and presenilin 1, and in Alzheimer's disease patients. Higher levels of nitro-triosephosphate isomerase (P < 0.05) were detected, by Western blot, in immunoprecipitates from hippocampus (9 individuals) and frontal cortex (13 individuals) of Alzheimer's disease patients, compared with healthy subjects (4 and 9 individuals, respectively). Triosephosphate isomerase nitrotyrosination decreases the glycolytic flow. Moreover, during its isomerase activity, it triggers the production of the highly neurotoxic methylglyoxal (n = 4; P < 0.05). The bioinformatics simulation of the nitration of tyrosines 164 and 208, close to the catalytic centre, fits with a reduced isomerase activity. Human embryonic kidney (HEK) cells overexpressing double mutant triosephosphate isomerase (Tyr164 and 208 by Phe164 and 208) showed high methylglyoxal production. This finding correlates with the widespread glycation immunostaining in Alzheimer's disease cortex and hippocampus from double transgenic mice overexpressing amyloid precursor protein and presenilin 1. Furthermore, nitro-triosephosphate isomerase formed large beta-sheet aggregates in vitro and in vivo, as demonstrated by turbidometric analysis and electron microscopy. Transmission electron microscopy (TEM) and atomic force microscopy studies have demonstrated that nitro-triosephosphate isomerase binds tau monomers and induces tau aggregation to form paired helical filaments, the characteristic intracellular hallmark of Alzheimer's disease brains. Our results link oxidative stress, the main etiopathogenic mechanism in sporadic Alzheimer's disease, via the production of peroxynitrite and nitrotyrosination of triosephosphate isomerase, to amyloid beta-peptide-induced toxicity and tau pathology.


Identification of a non-purple tartrate-resistant acid phosphatase: an evolutionary link to Ser/Thr protein phosphatases?
Tartrate-resistant acid phosphatases (TRAcPs), also known as purple acid phosphatases (PAPs), are a family of binuclear metallohydrolases that have been identified in plants, animals and fungi. The human enzyme is a major histochemical marker for the diagnosis of bone-related diseases. TRAcPs can occur as a small form possessing only the ~35 kDa catalytic domain, or a larger ~55 kDa form possessing both a catalytic domain and an additional N-terminal domain of unknown function. Due to its role in bone resorption the 35 kDa TRAcP has become a promising target for the development of anti-osteoporotic chemotherapeutics. A new human gene product encoding a metallohydrolase distantly related to the ~55 kDa plant TRAcP was identified and characterised. The gene product is found in a number of animal species, and is present in all tissues sampled by the RIKEN mouse transcriptome project. Construction of a homology model illustrated that six of the seven metal-coordinating ligands in the active site are identical to that observed in the TRAcP family. However, the tyrosine ligand associated with the charge transfer transition and purple color of TRAcPs is replaced by a histidine. The gene product identified here may represent an evolutionary link between TRAcPs and Ser/Thr protein phosphatases. Its biological function is currently unknown but is unlikely to be associated with bone metabolism.


Ketamine for Sickle Cell Vaso-Occlusive Crises: A Systematic Review.
Vaso-occlusive crisis (VOC) is one of the main causes of hospital admission in patients with sickle cell disease (SCD). Ketamine is often used as an adjuvant to opioids to control sickle cell crisis; however, there is a lack of evidence about its safety and efficacy for VOC in SCD patients. To synthesize evidence from published reports about the efficacy and safety of ketamine in the management of acute painful VOC in both pediatric and adult SCD patients. A systematic literature search of PubMed, Scopus, Web of Science, EBSCO and Cochrane Library was conducted, up to March 2019. Studies reporting the analgesic effects and side effects of ketamine in the management of acute painful VOC in pediatric and adult SCD patients were included. The primary outcome measure was improvement in pain scale, and the secondary outcomes were reduction in opioid utilization and side effects. Studies were narratively summarized in this review. Fourteen studies (with a total of 604 patients) were included in the final analysis. Several case reports and case series showed that ketamine significantly reduced pain scales and opioid utilization in both populations. The only randomized controlled trial available showed that ketamine was noninferior to morphine in reducing pain scores, but had a higher incidence of nonlife-threatening, reversible adverse effects. However, a retrospective study of 33 patients showed a higher pain score in the ketamine group with an acceptable short-term adverse effect. Ketamine has a potentially comparable efficacy with other opioids in reducing the pain during VOC in SCD patients. However, it also likely has a higher rate of transient adverse events. Owing to the lack of published randomized controlled trials, current evidence is not sufficient to confirm the safety and efficacy of ketamine. Future well-designed randomized controlled trials are strongly recommended.


31 P magnetic resonance spectroscopy in skeletal muscle: Experts' consensus recommendations.
Skeletal muscle phosphorus-31 31 P MRS is the oldest MRS methodology to be applied to in vivo metabolic research. The technical requirements of 31 P MRS in skeletal muscle depend on the research question, and to assess those questions requires understanding both the relevant muscle physiology, and how 31 P MRS methods can probe it. Here we consider basic signal-acquisition parameters related to radio frequency excitation, TR, TE, spectral resolution, shim and localisation. We make specific recommendations for studies of resting and exercising muscle, including magnetisation transfer, and for data processing. We summarise the metabolic information that can be quantitatively assessed with 31 P MRS, either measured directly or derived by calculations that depend on particular metabolic models, and we give advice on potential problems of interpretation. We give expected values and tolerable ranges for some measured quantities, and minimum requirements for reporting acquisition parameters and experimental results in publications. Reliable examination depends on a reproducible setup, standardised preconditioning of the subject, and careful control of potential difficulties, and we summarise some important considerations and potential confounders. Our recommendations include the quantification and standardisation of contraction intensity, and how best to account for heterogeneous muscle recruitment. We highlight some pitfalls in the assessment of mitochondrial function by analysis of phosphocreatine (PCr) recovery kinetics. Finally, we outline how complementary techniques (near-infrared spectroscopy, arterial spin labelling, BOLD and various other MRI and 1 H MRS measurements) can help in the physiological/metabolic interpretation of 31 P MRS studies by providing information about blood flow and oxygen delivery/utilisation. Our recommendations will assist in achieving the fullest possible reliable picture of muscle physiology and pathophysiology.


Chromosomal abnormalities in the human population: estimation of rates based on New Haven newborn study.
The incidence of gross chromosomal abnormality was measured in a large (4500), relatively unbiased sample of New Haven infants born during 1 year. The frequency of infants with abnormal chromosomal constitutions was 0.5 percent. For mothers over age 34, 1.5 percent of newborns were chromosomally abnormal. Only one in four of these infants could have been detected by phenotypic criteria alone. Methods are discussed whereby this fraction of the newborn population might be detected and possibly reduced.


Endoscopic ultrasound of pancreatic lesions.
Endoscopic ultrasound (EUS) is a well-established tool for the evaluation of pancreatic lesions. Due to the closer proximity of EUS to the pancreas, EUS offers a high sensitivity for detection of small pancreatic mass and is the preferred modality for obtaining tissue for diagnosis of pancreatic mass. Contrast-enhanced EUS and/or elastography provide additional information to the fundamental B-mode ultrasound images, leading to more accurate diagnosis. The aim of this video-article is to show the different steps in performing EUS on pancreatic lesions and to provide some tips and tricks to improve and facilitate the execution of EUS on pancreatic lesions.


Chromosomal influence on meiotic spindle assembly: abnormal meiosis I in female Mlh1 mutant mice.
In mouse oocytes, the first meiotic spindle is formed through the action of multiple microtubule organizing centers rather than a pair of centrosomes. Although the chromosomes are thought to play a major role in organizing the meiotic spindle, it remains unclear how a stable bipolar spindle is established. We have studied the formation of the first meiotic spindle in murine oocytes from mice homozygous for a targeted disruption of the DNA mismatch repair gene, Mlh1. In the absence of the MLH1 protein meiotic recombination is dramatically reduced and, as a result, the vast majority of chromosomes are present as unpaired univalents at the first meiotic division. The orientation of these univalent chromosomes at prometaphase suggests that they are unable to establish stable bipolar spindle attachments, presumably due to the inability to differentiate functional kinetochore domains on individual sister chromatids. In the presence of this aberrant chromosome behavior a stable first meiotic spindle is not formed, the spindle poles continue to elongate, and the vast majority of cells never initiate anaphase. These results suggest that, in female meiotic systems in which spindle formation is based on the action of multiple microtubule organizing centers, the chromosomes not only promote microtubule polymerization and organization but their attachment to opposite spindle poles acts to stabilize the forming spindle poles.


A DNA repair system specific for thermophilic Archaea and bacteria predicted by genomic context analysis.
During a systematic analysis of conserved gene context in prokaryotic genomes, a previously undetected, complex, partially conserved neighborhood consisting of more than 20 genes was discovered in most Archaea (with the exception of Thermoplasma acidophilum and Halobacterium NRC-1) and some bacteria, including the hyperthermophiles Thermotoga maritima and Aquifex aeolicus. The gene composition and gene order in this neighborhood vary greatly between species, but all versions have a stable, conserved core that consists of five genes. One of the core genes encodes a predicted DNA helicase, often fused to a predicted HD-superfamily hydrolase, and another encodes a RecB family exonuclease; three core genes remain uncharacterized, but one of these might encode a nuclease of a new family. Two more genes that belong to this neighborhood and are present in most of the genomes in which the neighborhood was detected encode, respectively, a predicted HD-superfamily hydrolase (possibly a nuclease) of a distinct family and a predicted, novel DNA polymerase. Another characteristic feature of this neighborhood is the expansion of a superfamily of paralogous, uncharacterized proteins, which are encoded by at least 20-30% of the genes in the neighborhood. The functional features of the proteins encoded in this neighborhood suggest that they comprise a previously undetected DNA repair system, which, to our knowledge, is the first repair system largely specific for thermophiles to be identified. This hypothetical repair system might be functionally analogous to the bacterial-eukaryotic system of translesion, mutagenic repair whose central components are DNA polymerases of the UmuC-DinB-Rad30-Rev1 superfamily, which typically are missing in thermophiles.


Occurrence, fate, behavior and ecotoxicological state of phthalates in different environmental matrices.
Because of their large and widespread application, phthalates or phthalic acid esters (PAEs) are ubiquitous in all the environmental compartements. They have been widely detected throughout the worldwide environment. Indoor air where people spend 65-90% of their time is also highly contaminated by various PAEs released from plastics, consumer products as well as ambient suspended particulate matter. Because of their widespread application, PAEs are the most common chemicals that humans are in contact with daily. Based on various exposure mechanisms, including the ingestion of food, drinking water, dust/soil, air inhalation and dermal exposure the daily intake of PAEs may reach values as high as 70 μg/kg/day. PAEs are involved in endocrine disrupting effects, namely, upon reproductive physiology in different species of fish and mammals. They also present a variety of additional toxic effects for many other species including terrestrial and aquatic fauna and flora. Therefore, their presence in the environment has attracted considerable attention due to their potential impacts on ecosystem functioning and on public health. This paper is a synthesis of the extensive literature data on behavior, transport, fate and ecotoxicological state of PAEs in environmental matrices: air, water, sediment, sludge, wastewater, soil, and biota. First, the origins and physicochemical properties of PAEs that control the behavior, transport and fate in the environment are reviewed. Second, the compilation of data on transport and fate, adverse environmental and human health effects, legislation, restrictions, and ecotoxicological state of the environment based on PAEs is presented.


Deep Learning for Chondrogenic Tumor Classification through Wavelet Transform of Raman Spectra.
The grading of cancer tissues is still one of the main challenges for pathologists. The development of enhanced analysis strategies hence becomes crucial to accurately identify and further deal with each individual case. Raman spectroscopy (RS) is a promising tool for the classification of tumor tissues as it allows us to obtain the biochemical maps of the tissues under analysis and to observe their evolution in terms of biomolecules, proteins, lipid structures, DNA, vitamins, and so on. However, its potential could be further improved by providing a classification system which would be able to recognize the sample tumor category by taking as input the raw Raman spectroscopy signal; this could provide more reliable responses in shorter time scales and could reduce or eliminate false-positive or -negative diagnoses. Deep Learning techniques have become ubiquitous in recent years, with models able to perform classification with high accuracy in most diverse fields of research, e.g., natural language processing, computer vision, medical imaging. However, deep models often rely on huge labeled datasets to produce reasonable accuracy, otherwise occurring in overfitting issues when the training data is insufficient. In this paper, we propose a chondrogenic tumor CLAssification through wavelet transform of RAman spectra (CLARA), which is able to classify with high accuracy Raman spectra obtained from bone tissues. CLARA recognizes and grades the tumors in the evaluated dataset with 97% accuracy by exploiting a classification pipeline consisting of the division of the original task in two binary classification steps, where the first is performed on the original RS signals while the latter is accomplished through the use of a hybrid temporal-frequency 2D transform.


Rayleigh-Ritz optimization of the structure function for a Kolmogorov power density and an approximate analytical solution for the two-point electric field correlation function.
Beginning with the exact Kolmogorov power spectrum for an extended random medium (such as interstellar space), the structure function is first expressed as an inverse Fourier-Bessel transform. An approximate series representation involving fractional powers of the transverse coordinate is then optimized by an equivalent Gaussian expansion up to two terms using the Rayleigh-Ritz technique. The domains of validity as well as the coherence profile of the resulting analytical solution of the two-point electric field correlation function are examined. It is shown that the correlation in both the transverse and longitudinal directions persists over distances up to a few orders of magnitude greater than that for a medium described by a Gaussian structure function.


The Evolution of the Opiate/Opioid Crisis in Cuyahoga County.
The United States continues to grapple with an epidemic of opiate/opioid drugs. This crisis initially manifested itself in the use and abuse of opioid pain relievers and has since seen an increase in illicit opiate/opioid drug use mortality. Cuyahoga County (metropolitan Cleveland) has been an area where the crisis has been particularly acute; this paper updates our previous experience. Most notable in the evolution of the drug epidemic has been an increase in mortality associated with fentanyl and an alarming rise in overall deaths, largely attributable to the emergence of fentanyl (a 64% increase in total overdose deaths from 2015 to 2016, with fentanyl increasing 324%). Fentanyl is a synthetic opioid with use in medical analgesia and anesthesia; however, most of the current supply is of clandestinely manufactured origin. Also of concern is the recent appearance of illicit fentanyl analogues, which are briefly described in this report. White males continue to be the most frequent overdose victims in the current crisis. A decrease of age appears to have taken place with the emergence of fentanyl with the most common age group being between 30 and 44 years of age. The majority of decedents are nonurban residents. Educationally, most of these decedents have a high school diploma or less schooling and a significant percentage consists of manual laborers. Medical examiners are an important source of information necessary to develop prevention and interdiction strategies. Challenges faced regarding adequate funding, instrumentation, and staffing are being felt.


Sensitive and Specific Detection of Xanthomonas oryzae pv. oryzae by Real-Time Bio-PCR Using Pathovar-Specific Primers Based on an rhs Family Gene.
The present study describes bio-polymerase chain reaction (PCR) assays to detect bacterial leaf blight caused by Xanthomonas oryzae pv. oryzae in rice. Successful control of X. oryzae. pv. oryzae requires a specific and reliable diagnostic tool. However, other X. oryzae pathovars are detected by currently available molecular and serological methods. In this study, SYBR Green real-time and conventional PCR primer sets were designed based on an rhs family gene of X. oryzae pv. oryzae KACC10331 because these genes are structurally diverse. The specificity of the primers was evaluated using purified DNA from 11 isolates of two X. oryzae pathovars, 21 other Xanthomonas species, and 4 other reference phytopathogenic bacteria and fungi. The assay was also able to detect at least two genome equivalents of cloned amplified target DNA using purified DNA. Thus, the SYBR Green real-time PCR-based method can be used for the rapid and specific detection of X. oryzae pv. oryzae and will potentially simplify and facilitate diagnosis and monitoring of this pathogen and guide plant disease management.


Effects of myo-inositol supplementation on oocyte's quality in PCOS patients: a double blind trial.
Polycystic ovary syndrome is the most common cause of chronic anovulation infertility in women in fertile period, and it's characterized by an increased production of androgens and estrogens. The administration of myo-inositol, a B complex vitamin, was associated with a decreased of serum testosterone and simultaneously, due to its ability to increase insulin sensitivity, women who received myo-inositol showed a great improvement of the ovulary function. Besides, the supplementation of inositol improves the oocytes' quality and increase the number of oocytes collected after ovarian stimulation in patients undergoing IVF (in vitro fertilization). The aim of this study is to determine the effects of myo-inositol on oocyte's quality on a sample of women with polycystic ovary syndrome. The patients were divided into two groups: patients of Group A in-took 2 g of myo-inositol + 200 microg of folic acid (Inofolic, LO.LI. Pharma, Rome, Italy) while Group B only 200 microg of folic acid, both groups took the treatment twice a day, continuously for 3 months. At the end of treatment, the number of follicles of diameter > 15 mm, visible at ultrasound during stimulation, and the number of oocytes recovered at the time of pick-ups were found to be significantly greater in the group treated with myo-inositol, so as the aver-age number of embryos transferred and embryo Score S1. Significantly reduced was the average number of immature oocytes (vesicles germ and degenerated oocytes) too. These data suggest that myoinositol may be useful in the treatment of PCOS patients undergoing ovulation induction, both for its insulin-sensitizing activity, and its role in oocyte maturation.


Integrated preclinical and clinical development of mTOR inhibitors in pancreatic cancer.
The purpose of this work was to determine the efficacy of inhibiting mammalian target of rapamycin (mTOR) in pancreatic cancer preclinical models and translate preclinical observations to the clinic. Temsirolimus (20 mg Kg(-1) daily) was administered to freshly generated pancreatic cancer xenografts. Tumour growth inhibition was determined after 28 days. Xenografts were characterised at baseline by gene expression and comparative genomic hybridisation. Patients with advanced, gemcitabine-resistant pancreatic cancer were treated with sirolimus (5 mg daily). The primary end point was 6-month survival rate (6mSR). Correlative studies included immunohistochemistry assessment of pathway expression in baseline tumours, drug pharmacokinetics (PKs), response assessment by FDG-PET and pharmacodynamic effects in peripheral-blood mononuclear cells (PBMCs). In all, 4 of 17 xenografts (23%) responded to treatment. Sensitive tumours were characterised by gene copy number variations and overexpression of genes leading to activation of the PI3K/Akt/mTOR pathway. Activation of p70S6K correlated with drug activity in the preclinical studies. Sirolimus was well tolerated in the clinic, showed predictable PKs, exerted pathway inhibition in post-treatment PBMCs and resulted in a 6mSR of 26%. No correlation, however, was found between activated p70S6K in tumour tissues and anti-tumour effects. Sirolimus activity in pancreatic cancer was marginal and not predicted by the selected biomarker.


Single-Gaussian-beam interaction with a dielectric microsphere: radiation forces, multiple internal reflections, and caustic structures.
We analyze the generation of radiation forces in the various internal reflections that occur when a tightly focused, unpolarized Gaussian beam interacts with a dielectric microsphere. The Euler transformation is used to compute the relevant axial-force vectors that are associated with the internal reflections rapidly. The net force is computed as the average of the forces generated by the s- and p-polarization components of the electric-field vector. The force characteristics are strongly dependent on the algebraic value of the index mismatch Δn at the sphere interface. For the two cases of Δn = 0.65, 0.32 with a sphere index n(2) = 1.65, the inclusion of the contributions from the second and third internal reflections after the magnitude of the effective force by at least 10%. For Δn < 0, the individual force contributions from internal reflections beyond the first are at least 4 orders of magnitude less than those of the first interaction. The caustic structures produced by the internal reflections are also profiled for both cases of Δn = ±0.32.


Surrogate testing suggests that chlorine dioxide gas exposure would not inactivate Ebola virus contained in environmental blood contamination.
The ability to decontaminate a room potentially containing the Ebola virus is important to healthcare facilities in the United States. Ebola virus remains viable in body fluids, a room that has housed a patient with Ebola virus disease must have all surfaces manually wiped with an approved disinfectant, which increases occupational exposure risk. This study evaluated the efficacy of gaseous chlorine dioxide inactivation of bacterial organisms in blood as Ebola virus surrogates and as the organisms used by the Nebraska Biocontainment Unit to provide the margin of safety for decontamination. Bacillus anthracis, Escherichia coli, Enterococcus faecalis, and Mycobacterium smegmatis blood suspensions that were exposed to ClO2 gas concentrations and exposure limits. The log reduction in Colony Forming Units (CFU) was determined for each bacterial blood suspension. Exposure parameters approximating industry practices for ClO2 environmental decontamination (360ppm concentration to 780 ppm-hrs exposure, 65% relative humidity) as well as parameters exceeding current practice (1116 ppm concentration to 1400 ppm-hrs exposure; 1342ppm concentration to 1487 ppm-hrs exposure) were evaluated. Complete inactivation was not achieved for any of the bacterial blood suspensions tested. Reductions were observed in concentrations of B. anthracis spores (1.3 -3.76 log) and E. faecalis vegetative cells (1.3 log) whereas significant reductions in vegetative cell concentrations for E. coli and M. smegmatis blood suspensions were not achieved. Our results showed that bacteria in the presence of blood were not inactivated using gaseous ClO2 decontamination. ClO2 decontamination alone should not be used for Ebola virus, but decontamination processes should first include manual wiping of potentially contaminated blood; especially for microorganisms as infectious as the Ebola virus.


Anti-PD-L1 Efficacy Can Be Enhanced by Inhibition of Myeloid-Derived Suppressor Cells with a Selective Inhibitor of PI3Kδ/γ.
Checkpoint inhibitors are relatively inefficacious in head and neck cancers, despite an abundance of genetic alterations and a T-cell-inflamed phenotype. One significant barrier to efficacy may be the recruitment of myeloid-derived suppressor cells (MDSC) into the tumor microenvironment. Here we demonstrate functional inhibition of MDSC with IPI-145, an inhibitor of PI3Kδ and PI3Kγ isoforms, which enhances responses to PD-L1 blockade. Combination therapy induced CD8+ T lymphocyte-dependent primary tumor growth delay and prolonged survival only in T-cell-inflamed tumor models of head and neck cancers. However, higher doses of IPI-145 reversed the observed enhancement of anti-PD-L1 efficacy due to off-target suppression of the activity of tumor-infiltrating T lymphocytes. Together, our results offer a preclinical proof of concept for the low-dose use of isoform-specific PI3Kδ/γ inhibitors to suppress MDSC to enhance responses to immune checkpoint blockade. Cancer Res; 77(10); 2607-19. ©2017 AACR.


Is there an association between metabolic syndrome and cognitive function in very old adults? The Newcastle 85+ Study.
To determine, using data from the Newcastle 85+ Study, whether there is an association between modern diagnostic criteria for metabolic syndrome (MetS) and cognitive function in very old adults (≥85) and whether inflammation, physical activity, or diabetes mellitus status affects this association. Longitudinal, population-based cohort study. Newcastle and North Tyneside, United Kingdom. Community-dwelling and institutionalized men and women recruited through general practices (N = 845). MetS was defined according to the National Cholesterol Education Program (NCEP) Adult Treatment Panel III (ATP III) criteria. Cross-sectional and prospective (up to 5 years of follow-up) associations between MetS and global cognitive function (assessed using the Mini-Mental State Examination (MMSE)) and between MetS and attention and episodic memory (assessed using the Cognitive Drug Research battery) were performed. MetS was not associated with cognitive function at baseline or cognitive change over time. Lack of association was not because MetS was predictive of subsequent mortality. Of the individual components of the MetS criteria, high blood pressure was associated with better cognitive function at baseline (MMSE: β (standard error (SE)) = -0.716 (0.152), P < .001), and low high-density lipoprotein cholesterol was associated with poorer global cognitive function at baseline (MMSE: 0.436 (0.131), P = .001). The association between MetS and cognitive decline, which has been described in younger populations (<75), was not apparent in this population of individuals aged 85 and older at baseline.


The Tudor domain protein Spindlin1 is involved in intrinsic antiviral defense against incoming hepatitis B Virus and herpes simplex virus type 1.
Hepatitis B virus infection (HBV) is a major risk factor for the development of hepatocellular carcinoma. HBV replicates from a covalently closed circular DNA (cccDNA) that remains as an episome within the nucleus of infected cells and serves as a template for the transcription of HBV RNAs. The regulatory protein HBx has been shown to be essential for cccDNA transcription in the context of infection. Here we identified Spindlin1, a cellular Tudor-domain protein, as an HBx interacting partner. We further demonstrated that Spindlin1 is recruited to the cccDNA and inhibits its transcription in the context of infection. Spindlin1 knockdown induced an increase in HBV transcription and in histone H4K4 trimethylation at the cccDNA, suggesting that Spindlin1 impacts on epigenetic regulation. Spindlin1-induced transcriptional inhibition was greater for the HBV virus deficient for the expression of HBx than for the HBV WT virus, suggesting that HBx counteracts Spindlin1 repression. Importantly, we showed that the repressive role of Spindlin1 is not limited to HBV transcription but also extends to other DNA virus that replicate within the nucleus such as Herpes Simplex Virus type 1 (HSV-1). Taken together our results identify Spindlin1 as a critical component of the intrinsic antiviral defense and shed new light on the function of HBx in HBV infection.


Fucoidan Derived from Fucus vesiculosus Inhibits the Development of Human Ovarian Cancer via the Disturbance of Calcium Homeostasis, Endoplasmic Reticulum Stress, and Angiogenesis.
Marine organisms are sources of several natural compounds with potential clinical use. However, only a few marine-based pharmaceuticals have been approved for use due to limited knowledge on their biological activities. Here, we identified the functional role of fucoidan extracted from Fucus vesiculosus on ovarian cancer. Fucoidan increased the death of ES-2 and OV-90 cells, through a reduction in proliferation, cell cycle arrest, releases of cytochrome c, reactive oxygen species (ROS) generation, and endoplasmic reticulum (ER) stress. Additionally, fucoidan increased the concentration of cytosolic and mitochondrial calcium in both cells. The decrease of cell proliferation was controlled by the inactivation of PI3K and MAPK signaling cascades in ES-2 and OV-90 cells. In a toxicity assay with normal zebrafish larvae, fucoidan did not induce toxicity, cardiotoxicity, development, kinesis, and apoptosis at different concentrations. However, it disrupted tumor formation and vascular development in a zebrafish xenograft model and angiogenesis transgenic (Tg, fli1-eGFP) model, respectively. Collectively, the results indicate that fucoidan may be a novel pharmaceutical for the management of human ovarian cancer.


Patients' preferences for labor analgesic counseling: A qualitative analysis.
The decision to use, or not use, neuraxial labor analgesia is complex, with both maternal and fetal considerations. Fears and concerns about neuraxial analgesia may influence analgesic decision-making. Little is known about patients' preferences for analgesic counseling. Therefore, the objectives of this qualitative study were to evaluate the sources of information used by patients, the timing and content of antepartum counseling about labor analgesia, and patients' preferences for such counseling. A semi-structured interview guide was developed. Survey domains included the sources of information used by patients, the timing and content of antenatal counseling, patients' perceptions of intrapartum analgesic counseling, and their preferences for analgesic counseling. Interviews occured on postpartum day 1. Interview transcripts were analyzed using content analysis. The interviews of 45 women were analyzed. The Internet was the most frequently mentioned source of information (44%). The majority of women (78%) discussed analgesic options with their obstetric providers before delivery. All women were counseled intrapartum by an anesthesiologist, but several women commented on the difficulty of concentrating while laboring. Seventy-three percent of women stated that they would want counseling on analgesic options from their obstetric provider during the second or third trimester. Women prefer to be counseled about labor analgesia by their obstetric providers, as they have an established relationship. Patients would prefer the counseling to occur during pregnancy, before the onset of labor. However, given the frequent use of the Internet, the content and quality of online materials should also be evaluated.


Identifying geographic disparities in the early detection of breast cancer using a geographic information system.
Identifying communities with lower rates of mammography screening is a critical step to providing targeted screening programs; however, population-based data necessary for identifying these geographic areas are limited. This study presents methods to identify geographic disparities in the early detection of breast cancer. Data for all women residing in Dane County, Wisconsin, at the time of their breast cancer diagnosis from 1981 through 2000 (N = 4769) were obtained from the Wisconsin Cancer Reporting System (Wisconsin's tumor registry) by ZIP code of residence. Hierarchical logistic regression models for disease mapping were used to identify geographic differences in the early detection of breast cancer. The percentage of breast cancer cases diagnosed in situ (excluding lobular carcinoma in situ) increased from 1.3% in 1981 to 11.9% in 2000. This increase, reflecting increasing mammography use, occurred sooner in Dane County than in Wisconsin as a whole. From 1981 through 1985, the proportion of breast cancer diagnosed in situ in Dane county was universally low (2%-3%). From 1986 through 1990, urban and suburban ZIP codes had significantly higher rates (10%) compared with rural ZIP codes (5%). From 1991 through 1995, mammography screening had increased in rural ZIP codes (7% of breast cancer diagnosed in situ). From 1996 through 2000, mammography use was fairly homogeneous across the entire county (13%-14% of breast cancer diagnosed in situ). The percentage of breast cancer cases diagnosed in situ increased in the state and in all areas of Dane County from 1981 through 2000. Visual display of the geographic differences in the early detection of breast cancer demonstrates the diffusion of mammography use across the county over the 20-year period.


Demographic and social characteristics of adults with cystic fibrosis in the United Kingdom.
To obtain information about social and demographic characteristics and lifestyle of adult patients with cystic fibrosis, including those who do not attend major specialist clinics. Confidential self completion postal questionnaire to adult patients with cystic fibrosis, asking about social and demographic characteristics, social class and occupation, employment, education, insurance and social security benefits, symptom severity, and medical care. National association for adults with cystic fibrosis. 1052 adult members of the Association of Cystic Fibrosis Adults UK, accounting for 68% of those with cystic fibrosis in the United Kingdom population over 16 years of age and over 80% of those over 25 in June 1990. The response rate was 82% (397 women, 423 men). Most adults with cystic fibrosis were found to be living fulfilling lives into adulthood. Significantly fewer men were married or cohabiting than women (110 (26%) men, 175 (44%) women). 420 (55%) responders were working, and of these 235 (56%) had less than two weeks' sick leave a year. Half of those not employed gave ill health as the reason. Revealing that they had cystic fibrosis at job interviews reduced likelihood of being employed for those with mild to moderate disease. People with cystic fibrosis had been less successful than the general population in achieving O level or equivalent qualifications, but more successful in achieving A level or higher qualifications. Achievement of any qualifications enhanced employment prospects irrespective of disease severity. Contrary to an image of chronic ill health and disability, a high proportion of adults with cystic fibrosis are living full and productive lives.


Systemic RALA/iNOS Nanoparticles: A Potent Gene Therapy for Metastatic Breast Cancer Coupled as a Biomarker of Treatment.
This study aimed to determine the therapeutic benefit of a nanoparticular formulation for the delivery of inducible nitric oxide synthase (iNOS) gene therapy in a model of breast cancer metastasis. Nanoparticles comprising a cationic peptide vector, RALA, and plasmid DNA were formulated and characterized using a range of physiochemical analyses. Nanoparticles complexed using iNOS plasmids and RALA approximated 60 nm in diameter with a charge of 25 mV. A vector neutralization assay, performed to determine the immunogenicity of nanoparticles in immunocompetent C57BL/6 mice, revealed that no vector neutralization was evident. Nanoparticles harboring iNOS plasmids (constitutively active cytomegalovirus [CMV]-driven or transcriptionally regulated human osteocalcin [hOC]-driven) evoked iNOS protein expression and nitrite accumulation and impaired clonogenicity in the highly aggressive MDA-MB-231 human breast cancer model. Micrometastases of MDA-MB-231-luc-D3H1 cells were established in female BALB/c SCID mice by intracardiac delivery. Nanoparticulate RALA/CMV-iNOS or RALA/hOC-iNOS increased median survival in mice bearing micrometastases by 27% compared with controls and also provoked elevated blood nitrite levels. Additionally, iNOS gene therapy sensitized MDA-MB-231-luc-D3H1 tumors to docetaxel treatment. Studies demonstrated that systemically delivered RALA-iNOS nanoparticles have therapeutic potential for the treatment of metastatic breast cancer. Furthermore, detection of nitrite levels in the blood serves as a reliable biomarker of treatment.


Review of the molecular modelling studies of the cytochrome P-450 estrogen synthetase enzyme, aromatase.
The biosynthesis of the family of female hormones, the estrogens, is mediated by the cytochrome P-450 based enzyme Aromatase (AR), an enzyme which has been the focus of extensive research for some time and has resulted in the synthesis of numerous inhibitory compounds with varying structural properties. The crystal structure of this membrane-bound enzyme has not been determined, although that of the bacterial P-450 camphor hydroxylase enzyme has, and the latter has been used for homology modelling of AR and other related enzymes. Several workers have attempted to obtain an approximate model of the active site of AR through the use of various molecular modelling techniques using inhibitors of this enzyme, as well as a combination of experiments involving active site mutagenesis of specific areas of the P-450 backbone together with molecular modelling. In this report these studies are reviewed. In summary, although the crystal structure of AR has yet to be determined, the use of molecular modelling studies has provided useful information in the design and synthesis of its inhibitors.


Once-daily fluticasone furoate (FF)/vilanterol reduces risk of severe exacerbations in asthma versus FF alone.
Combination therapy with an inhaled corticosteroid (ICS) and long-acting β2 agonist (LABA) is recommended for patients with asthma symptomatic on ICS alone. However, there is ongoing debate regarding the risk-benefit ratio of using LABA in asthma. To evaluate the effect of the addition of a novel LABA, vilanterol (VI), to a once-daily ICS, fluticasone furoate (FF), on the risk of severe asthma exacerbations in patients with uncontrolled asthma. This randomised double-blind comparative study of variable duration (≥ 24-78 weeks) was designed to finish after 330 events (each patient's first on-treatment severe asthma exacerbation). 2019 patients with asthma aged ≥ 12 years with ≥ 1 recorded exacerbation within 1 year were randomised and received FF/VI 100/25 μg or FF 100 μg, administered once daily in the evening. The primary endpoint was time to first severe exacerbation; secondary endpoints were rate of severe asthma exacerbations per patient per year and change in trough evening forced expiratory volume in 1 s (FEV1) from baseline. Compared with FF, FF/VI delayed the time to first severe exacerbation (HR 0.795, 95% CI 0.642 to 0.985) and reduced the annualised rate of severe exacerbations (rate reduction 25%, 95% CI 5% to 40%). Significantly greater improvements in trough FEV1 (p<0.001) were observed with FF/VI than with FF at weeks 12, 36, 52 and at endpoint. Both treatments were well tolerated with similar rates of treatment-related adverse events and on-treatment serious adverse events. Once-daily FF/VI reduced the risk of severe asthma exacerbations and improved lung function compared with FF alone, with good tolerability and safety profile in adolescents and adults with asthma currently receiving ICS. NCT01086384.


What the brain 'Likes': neural correlates of providing feedback on social media.
Evidence increasingly suggests that neural structures that respond to primary and secondary rewards are also implicated in the processing of social rewards. The 'Like'-a popular feature on social media-shares features with both monetary and social rewards as a means of feedback that shapes reinforcement learning. Despite the ubiquity of the Like, little is known about the neural correlates of providing this feedback to others. In this study, we mapped the neural correlates of providing Likes to others on social media. Fifty-eight adolescents and young adults completed a task in the MRI scanner designed to mimic the social photo-sharing app Instagram. We examined neural responses when participants provided positive feedback to others. The experience of providing Likes to others on social media related to activation in brain circuity implicated in reward, including the striatum and ventral tegmental area, regions also implicated in the experience of receiving Likes from others. Providing Likes was also associated with activation in brain regions involved in salience processing and executive function. We discuss the implications of these findings for our understanding of the neural processing of social rewards, as well as the neural processes underlying social media use.


Natural course of subjects with elevated liver tests and normal liver histology.
Liver biopsy (LB) is performed if non-invasive work-up of liver disease is inconclusive. The examination of liver tissue occasionally reveals normal histology. Long-term follow-up of such patients has not been performed. We identified a total 70 subjects from our LB database with elevated liver tests and normal liver histology after a mean of 90.5 ± 52.3 (range 15-216) months and conducted reassessment of medical history, physical examination, laboratory testing, ultrasound, transient elastography and LB if indicated. At follow-up examination, 15 (7 females (f)/8 males (m); 21.4%) subjects had normal liver tests and no further evidence of liver disease. A subset of 37 (29 f/8 m; 52.9%) subjects had persistently elevated liver tests without evidence indicating progressive liver disease but the cause thereof remained unexplained also at the follow-up visit. Three (0 f/3 m; 4.3%) subjects had consumed excessive alcohol with indicators of alcoholic liver disease. Eleven subjects (4 f/7 m; 15.7%) had developed steatosis on ultrasound examination along with weight gain and/or biochemical features of the metabolic syndrome. In addition, three (2 f/1 m) patients developed autoimmune hepatitis, one female presented with primary biliary cirrhosis. One male was diagnosed with cholangiocellular carcinoma 3 months after the initial evaluation. The clinical course of most patients was benign, but in approximately 20% of the subjects a liver disease developed. Particular attention should be given to autoimmune liver diseases in subjects with positive autoantibodies. In addition, lifestyle factors such as weight gain and alcohol consumption were associated with the manifestation of liver diseases.


The Interrater Reliability of the Autism Diagnostic Interview-Revised (ADI-R) in Clinical Settings.
The Autism Diagnostic Interview-Revised (ADI-R) is considered a first choice assessment tool in autism spectrum disorder. Nevertheless, despite its wide use in psychiatric practice and recommendations by various clinical guidelines, its interrater reliability has predominantly been confirmed in research settings by specially trained, research reliability interviewers. The reliability of ADI-R assessments among clinicians has not yet been established. Therefore, this study examined the spontaneous interrater reliability of the ADI-R in a naturalistic clinical multicenter setting. Ten video-recorded ADI-R administrations were rated by 5 different raters each from a pool of 11 raters affiliated to 8 different clinical sites. The interrater reliability for the 12 diagnostic criteria operationalizing autism spectrum disorders according to DSM-IV/ICD-10 in the ADI-R algorithms ranged between G(q,k) (analogous to intraclass correlations) = 0.96 and 0.99 for reciprocal social interaction, 0.96 and 1.00 for communication, and 0.91 and 0.97 for repetitive and restricted behavior. Reliability of diagnostic classification was ĸCohen 0.83. The findings endorse the psychometric properties of ADI-R in terms of interrater reliability previously reported from research settings and support their generalization to common clinical settings. Limitations of this study include an unbalanced sample composition.


Lp(a) glycoprotein phenotypes. Inheritance and relation to Lp(a)-lipoprotein concentrations in plasma.
The Lp(a) lipoprotein represents a quantitative genetic trait. It contains two different polypeptide chains, the Lp(a) glycoprotein and apo B-100. We have demonstrated the Lp(a) glycoprotein directly in human sera by sodium dodecyl sulfate-gel electrophoresis under reducing conditions after immunoblotting using anti-Lp(a) serum and have observed inter- and intraindividual size heterogeneity of the glycoprotein with apparent molecular weights ranging from approximately 400,000-700,000 D. According to their relative mobilities compared with apo B-100 Lp(a) patterns were categorized into phenotypes F (faster than apo B-100), B (similar to apo B-100), S1, S2, S3, and S4 (all slower than apo B-100), and into the respective double-band phenotypes. Results from neuraminidase treatment of isolated Lp(a) glycoprotein indicate that the phenotypic differences do not reside in the sialic acid moiety of the glycoprotein. Family studies are compatible with the concept that Lp(a) glycoprotein phenotypes are controlled by a series of autosomal alleles (Lp[a]F, Lp[a]B, Lp[a]S1, Lp[a]S2, Lp[a]S3, Lp[a]S4, and Lp[a]0) at a single locus. Comparison of Lp(a) plasma concentrations in different phenotypes revealed a highly significant association of phenotype with concentration. Phenotypes B, S1, and S2 are associated with high and phenotypes S3 and S4 with low Lp(a) concentrations. This suggests that the same gene locus is involved in determining Lp(a) glycoprotein phenotypes and Lp(a) lipoprotein concentrations in plasma and is the first indication for structural differences underlying the quantitative genetic Lp(a)-trait.


Nucleotide sequence of the gelatinase gene (gelE) from Enterococcus faecalis subsp. liquefaciens.
The gene coding for gelatinase (also called metalloendopeptidase II; microbial proteinase, EC 22.214.171.124) of Enterococcus faecalis subsp. liquefaciens strain OG1-10 was cloned in an Escherichia coli-Enterococcus shuttle vector, and its nucleotide sequence was determined. The DNA sequence encodes one large open reading frame (ORF) with 509 amino acid residues. The ORF contains a signal sequence in its N-terminal region, whereas the N-terminal amino acid sequence determined from the purified extracellular proteinase starts at residue 192 deduced from the ORF. This implies that the gelatinase is synthesized as a prepropolypeptide or prezymogen. The mature gelatinase contains 318 amino acid residues (molecular weight, 34,582) and has significant homology with neutral proteinases from Bacillus species and elastase from Pseudomonas aeruginosa.


Improving foodborne illness prevention among transplant recipients.
The U.S. Department of Agriculture developed a food safety brochure to educate transplant recipients and their caregivers about the risk of contracting foodborne illnesses and safe food-handling prevention practices. Qualitative research was conducted with transplant recipients and caregivers to collect information on participants' food safety concerns, knowledge, and practices; changes in food safety knowledge and practices after receiving the food safety brochure; preferred communication channels and dissemination strategies for delivering the brochure; and evaluation of the brochure. Many participants received food safety information from a health care provider, but the content and format varied by institution. Most participants are not following recommended practices to cook and chill food safely, and many participants consume high-risk foods associated with foodborne illnesses. After reading the brochure, many participants made or plan to make at least one food safety recommendation. The research findings were used to refine the brochure before nationwide distribution.


Medical care use and expenditure among children and youth in the United States: analysis of a national probability sample.
Good access to health care for all US children and youth remains an important social policy goal. Recent patterns of access as reflected in the presence of regular care sources, health care use, health insurance coverage, and expenditures for medical care are described and analyzed using the subsample of all children 0 to 18 years of age from the 1980 National Medical Care Utilization and Expenditure Survey. Data from the survey indicate that in 1980, 92% of US children and youth had a regular care source and the same percentage were covered for the full year or part of the year by some form of public or private health insurance. However, use rates and patterns of expenditure continued to differ dramatically according to family background factors, particularly race, ethnicity, poverty status, and location of residence. These differences are analyzed and comparative data are presented for groups of children from various sociodemographic groups. Minority-group and near-poor children were found to be at highest risk for limited utilization of services and inadequate insurance coverage.


Role of ROS and RNS Sources in Physiological and Pathological Conditions.
There is significant evidence that, in living systems, free radicals and other reactive oxygen and nitrogen species play a double role, because they can cause oxidative damage and tissue dysfunction and serve as molecular signals activating stress responses that are beneficial to the organism. Mitochondria have been thought to both play a major role in tissue oxidative damage and dysfunction and provide protection against excessive tissue dysfunction through several mechanisms, including stimulation of opening of permeability transition pores. Until recently, the functional significance of ROS sources different from mitochondria has received lesser attention. However, the most recent data, besides confirming the mitochondrial role in tissue oxidative stress and protection, show interplay between mitochondria and other ROS cellular sources, so that activation of one can lead to activation of other sources. Thus, it is currently accepted that in various conditions all cellular sources of ROS provide significant contribution to processes that oxidatively damage tissues and assure their survival, through mechanisms such as autophagy and apoptosis.


Bot fly parasitism (Rogenhofera bonaerensis) (Diptera, Cuterebridae) in the pampean grassland mouse (Akodon azarae), in Argentina.
Seasonality and impact of parasitism by the larvae of Rogenhofera bonaerensis (Diptera: Cuterebridae) in pampean grassland mouse (Akodon azarae, Rodentia, Cricetidae) populations were studied in grasslands and cropfield borders near Diego Gaynor (Buenos Aires Province, Argentina), from November 1985 to December 1986. Trapped mice infections ranged from one larva (77%) to four (4%) per host; larvae occurred on tails (75.7%), backs (17.1%), thighs (5.7%), and abdomen (1.4%). Prevalence of larvae occurred in late spring (November and December) (35%), and in fall (March to June) (20%), with a peak of 25% during April and May. In spring these larvae affected 46% of resident and 19% of transient individuals; however, no differences were found between residents and transients in spring or fall. There was no significant relationship between mouse population density and prevalence of parasitism (r = -0.56). The annual percentage of infected rodents changed with age (body size classes): adults (37%) greater than subadults (25%) greater than juveniles (16%); with no differences between the sexes. Similar results were recorded in the spring, but there were no differences in the fall. Significant differences were not detected between stable (22% infected) and disturbed habitats (14% infected). During winter uninfected mice survived better than those infected during the previous fall. In contrast, the summer survival among spring infected mice and noninfected mice was similar. Parasitism showed no effects on reproductive activity and on home range size. Larvae showed strong host specificity for A. azarae (99%). Only one case was recorded in the long-tailed mouse (Oligoryzomys flavescens), and none in the white paunch mouse (Calomys laucha), both species sympatric with A. azarae.(ABSTRACT TRUNCATED AT 250 WORDS)


Contribution of the infection-associated complement regulator-acquiring surface protein 4 (ErpC) to complement resistance of Borrelia burgdorferi.
Borrelia burgdorferi evades complement-mediated killing by interacting with complement regulators through distinct complement regulator-acquiring surface proteins (CRASPs). Here, we extend our analyses to the contribution of CRASP-4 in mediating complement resistance of B. burgdorferi and its interaction with human complement regulators. CRASP-4 (also known as ErpC) was immobilized onto magnetic beads and used to capture proteins from human serum. Following Western blotting, factor H (CFH), CFH-related protein 1 (CFHR1), CFHR2, and CFHR5 were identified as ligands of CRASP-4. To analyze the impact of native CRASP-4 on mediating survival of serum-sensitive cells in human serum, a B. garinii strain was generated that ectopically expresses CRASP-4. CRASP-4-producing bacteria bound CFHR1, CFHR2, and CFHR5 but not CFH. In addition, transformed spirochetes deposited significant amounts of lethal complement components on their surface and were susceptible to human serum, thus indicating that CRASP-4 plays a subordinate role in complement resistance of B. burgdorferi.