Mislabeling of elutriation medium: a demonstration of the need to improve the quality control of reagents produced for cell processing.
The processing of hematopoietic cells to remove red cells, T-lymphocytes, or malignant cells has become common. However, few of the reagents used are produced specifically for marrow processing and approved by the Food and Drug Administration. A labeling error is reported in a commercially manufactured, research-grade reagent used in an elutriation procedure to deplete T cells from mononuclear cells obtained from bone marrow for allogeneic transplantation. The elutriation medium used in the procedure had been labeled as containing 0.9-percent sodium chloride, 0.1-percent dextrose, and 0.3 mM EDTA, but it actually contained 3.0 mM EDTA. When this medium was not available from the manufacturer, it was prepared in this hospital, according to the stated formulation, with 0.3 mM EDTA. When marrows from two unrelated donors were processed with this 0.3 mM EDTA-containing medium, cell loss due to aggregation occurred, and, in one recipient, the marrow did not engraft. After the error in the medium composition was discovered, marrow was collected a second time from the same donor. The marrow was processed with medium containing 3.0 mM EDTA, and no cell aggregation was observed. Food and Drug Administration-approved reagents, prepared specifically for use in cell processing and produced according to good manufacturing practices, should be used whenever possible.


Long-Term Cognitive Functioning and Psychological Well-Being in Surgically Treated Patients with Low-Grade Glioma.
The aim of this work is to provide an in-depth investigation of the impact of low-grade gliomas (LGG) and their surgery on patients' cognitive and emotional functioning and well-being, carried out via a comprehensive and multiple-measure psychological and neuropsychological assessment. Fifty surgically treated patients with LGG were evaluated 40 months after surgery on their functioning over 6 different cognitive domains, 3 core affective/emotional aspects, and 3 different psychological well-being measures to obtain a clearer picture of the long-term impact of illness and surgery on their psychological and relational world. Close relatives were also involved to obtain an independent measure of the psychological dimensions investigated. Cognitive status was satisfactory, with only mild short-term memory difficulties. The affective and well-being profile was characterized by mild signs of depression, good satisfaction with life and psychological well-being, and good personality development, with patients perceiving themselves as stronger and better persons after illness. However, patients showed higher emotional reactivity, and psychological well-being measures were negatively affected by epileptic burden. Well-being was related to positive affective/emotional functioning and unrelated to cognitive functioning. Good agreement between patients and relatives was found. In the long-term, patients operated on for LGG showed good cognitive functioning, with no significant long-term cognitive sequelae for the extensive surgical approach. Psychologically, patients appear to experience a deep psychological change and maturation, closely resembling that of so-called posttraumatic growth, which, to our knowledge, is for the first time described and quantified in patients with LGG.


The Impact of the COVID-19 Pandemic on Ophthalmic Care at an Eye-Specific Emergency Department in an Outbreak Hotspot.
During the COVID-19 pandemic, there is growing concern that patients are forgoing necessary care. Emergency departments (ED) represent an important site of eye care. We analyzed patterns of ED visits at an eye-specific ED since the declaration of the public health crisis. In this retrospective, cross-sectional single center study, medical records of 6744 patients who presented to the Massachusetts Eye and Ear ED between March 1st and April 30th in 2018, 2019, and 2020 were studied. The primary outcome measures were total volume of ED visits, proportion of urgent ED visits, and proportion of surgical visits. Overall, the median number of daily visits to the ED decreased by 18 visits per day since the declaration of public health guidelines (interquartile range, 9-24, p < 0.001). This accounted for a 32% decrease in the total volume of ED visits in 2020 compared to prior years during the study period (p < 0.001). There was a 9% increase in the proportion of primary diagnoses considered urgent (p = 0.002). The proportion of visits requiring urgent surgery increased by 39% (p = 0.004). The total number of eye-specific ED visits dropped compared to prior years while the proportion of urgent visits increased. Patients were likely more reluctant to seek eye care, deferring less urgent evaluation.


Assessment and management of upper and lower extremity impairment and disability.
The assessment and management of occupational and nonoccupational injuries and illnesses of the extremities require knowledge of the same concepts of impairment and disability that are used when evaluating or treating problems of other body systems. Dr. Talmage discusses the need for an accurate and objectively verifiable diagnosis and focuses on upper extremity pain in workers and the term tendinitis'. The concept of maximal medical improvement is considered, with examples of knee injuries to the menisci and localized articular cartilage defects. Causation controversies are explored using carpal tunnel syndrome as the example. The impairment rating process is illustrated by use of the AMA Guides system. A "checklist" to insure completeness of the examination of extremity problems is offered. Disability assessment and management is discussed, with an emphasis on the difference between abilities (capacities) and risk-based restrictions. The author concludes with thoughts on how to minimize disability.


Cultural background influences implicit but not explicit sense of agency for the production of musical tones.
The sense of agency is suggested to occur at both low and high levels by the involvement of sensorimotor processes and the contribution of retrospective inferences based on contextual cues. In the current study, we recruited western and non-western participants and examined the effect of pleasantness of action outcomes on both feeling of control ratings and intentional binding which refers to the perceived compression of the temporal delay between actions and outcomes. We found that both western and non-western groups showed greater feeling of control ratings for the consonant (pleasant) compared to dissonant (unpleasant) outcomes. The intentional binding effect, on the other hand, was stronger for the consonant compared to dissonant outcomes in the western group only. We discuss the results in relation to how cultural background might differentially influence the effect of outcome pleasantness on low and high levels of the sense of agency.


Understanding the Consumption of Antimicrobial Resistance-Related Content on Social Media: Twitter Analysis.
Antimicrobial resistance (AMR) is one of the most pressing concerns in our society. Today, social media can function as an important channel to disseminate information about AMR. The way in which this information is engaged with depends on a number of factors, including the target audience and the content of the social media post. The aim of this study is to better understand how AMR-related content is consumed on the social media platform Twitter and to understand some of the drivers of engagement. This is essential to designing effective public health strategies, raising awareness about antimicrobial stewardship, and enabling academics to effectively promote their research on social media. We took advantage of unrestricted access to the metrics associated with the Twitter bot @AntibioticResis, which has over 13,900 followers. This bot posts the latest AMR research in the format of a title and a URL link to the PubMed page for an article. The tweets do not contain other attributes such as author, affiliation, or journal. Therefore, engagement with the tweets is only affected by the words used in the titles. Using negative binomial regression models, we measured the impact of pathogen names in paper titles, academic attention inferred from publication counts, and general attention estimated from Twitter on URL clicks to AMR research papers. Followers of @AntibioticResis consisted primarily of health care professionals and academic researchers whose interests comprised mainly AMR, infectious diseases, microbiology, and public health. Three World Health Organization (WHO) critical priority pathogens-Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacteriaceae-were positively associated with URL clicks. Papers with shorter titles tended to have more engagements. We also described some key linguistic characteristics that should be considered when a researcher is trying to maximize engagement with their publication. Our finding suggests that specific pathogens gain more attention on Twitter than others and that the levels of attention do not necessarily correspond to their status on the WHO priority pathogen list. This suggests that more targeted public health strategies may be needed to raise awareness about AMR among specific pathogens. Analysis of follower data suggests that in the busy schedules of health care professionals, social media offers a fast and accessible gateway to staying abreast of the latest developments in this field.


Insights into membrane-bound presenilin 2 from all-atom molecular dynamics simulations.
Presenilins 1 and 2 (PS1 or PS2) are main genetic risk factors of familial Alzheimer's disease (AD) that produce the β-amyloid (Aβ) peptides and also have important stand-alone functions related to, e.g. calcium signaling. Most work so far has focused on PS1, but humans carry both PS1 and PS2, and mutations in both cause AD. Here, we develop a computational model of PS2 in the membrane to address the question how pathogenic PS2 mutations affect the membrane-embedded protein. The models are based on cryo-electron microscopy structures of PS1 translated to PS2, augmented with missing residues and a complete all-atom membrane-water system, and equilibrated using three independent 500-ns simulations of molecular dynamics with a structure-balanced force field. We show that the nine-transmembrane channel structure is substantially controlled by major dynamics in the hydrophilic loop bridging TM6 and TM7, which functions as a 'plug' in the PS2 membrane channel. TM2, TM6, TM7 and TM9 flexibility controls the size of this channel. We find that most pathogenic PS2 mutations significantly reduce stability relative to random mutations, using a statistical ANOVA test with all possible mutations in the affected sites as a control. The associated loss of compactness may also impair calcium affinity. Remarkably, similar properties of the open state are known to impair the binding of substrates to γ-secretase, and we thus argue that the two mechanisms could be functionally related.Communicated by Ramaswamy H. Sarma.


Impact of a public cholesterol screening program.
The National Cholesterol Education Program (NCEP) has endorsed physician case finding as the primary method to detect individuals with elevated cholesterol levels. Despite this recommendation, promotional and for-profit public screening programs have flourished. We surveyed participants of a mall-based cholesterol screening program 1 year after their screening. Sixty-four percent of those screened had not previously known their cholesterol levels. Those who were newly screened were less likely to benefit from this testing than the general public, since they were older (mean age, 55.3 years), more likely to be female (67.4%), and nonsmokers (88%). Screenees had excellent recall of their cholesterol level (mean absolute reporting error, 0.24 mmol/L [9 mg/dL]) and a good understanding of cholesterol as a coronary heart disease risk. Those with elevated cholesterol levels reported high distress from screening but no reduction in overall psychosocial well-being and an actual decrease in absenteeism. Only 53.7% of all who were advised to seek follow-up because of an elevated screening value had done so within the year following the screening program. However, of those with values greater than 6.2 mmol/L (240 mg/dL), 68% had sought follow-up. Many of those who participate in public screening programs have been previously tested, fall into low-benefit groups, or fail to comply with recommended follow-up. We therefore conclude that cholesterol screening programs of the type now commonly offered are unlikely to contribute greatly to the national efforts to further reduce coronary heart disease.


Oxy-PAHs: occurrence in the environment and potential genotoxic/mutagenic risk assessment for human health.
Structurally modified polycyclic aromatic hydrocarbons (PAHs) such as nitrated PAHs (nitro-PAHs) and oxygenated PAHs (oxy-PAHs) can be incriminated in the total toxicity of polycyclic aromatic compounds (PACs) fraction in the environment. Compared to nitro-PAHs, oxy-PAHs have been poorly studied. Oxy-PAHs covers compounds with different moieties such as polycyclic aromatic ketones (PAKs) and polycyclic aromatic quinones (PAQs). In this review, we have compiled exhaustively all the data available on the sources, the fate, and the occurrence of oxy-PAHs focusing on the most ubiquitous ones in the environment, ie PAKs and PAQs. Data concerning their genotoxicity, mutagenicity and tumor promotion potential for humans are also provided based on the mode-of-action analysis framework. Mutagenicity results based on the limited number of oxy-PAHs tested, are unequivocal on the concern they represent. Their omission in mutagenic/carcinogenic risk has caused a dramatic underestimation of cancer risk. On the basis of environmental and genotoxicological data, we suggest prioritized 4 major oxy-PAHs molecules in ecotoxicological and toxicological studies, namely 6 H-benzo[cd]pyren-6-one (BPO), 7,12-benz[a]anthracenequinone (BAQ), 5,12-naphthacenequinone (NCQ) and 11 H-benzo[b]fluoren-11-one (B[b]FO). We also propose to develop biomarkers of exposure and/or risk for these compounds, for example by quantification of DNA adducts.


Synthesis of 6-aziridinylbenzimidazole derivatives and their in vitro antitumor activities.
In search for new antitumor agents, twelve 6-aziridinylbenzimidazole derivatives were synthesized and their cytotoxicities were tested against three cancer cell lines (mouse lymphocytic leukemia P388 and B16, and human gastric carcinoma SNU-16). From 4-amino-3-nitrotoluene as the starting material, 2-(acetoxymethyl)benzimidazoles (5a-d) were obtained by Phillips reaction. These benzimidazoles were then reacted with Fremy's salt to give a mixture of three 2-(acetoxymethyl) (8a-c) and four 2-(hydroxymethyl)benzimidazole-4,7-diones (9a-d). Addition of these quinones with aziridine afforded 6-aziridinyl-2-(acetoxymethyl) (10a-c) and 6-aziridinyl-2-(hydroxymethyl)benzimidazole-4,7-diones (11a-d). Utilizing 2-(hydroxymethyl)benzimidazole-4,7-diones (9b,d), esters 10d and 13e-h were prepared by the sequential reactions of esterification and addition. The synthesized compounds show potent cytotoxicity against all of three cell lines tested. The cytotoxicities of 10a-d or 11a-d against SNU-16 were superior to those of 13e-h, and were equal to or slightly higher than that of mitomycin C. Compounds 11a-d were slightly more cytotoxic than 10a-d in all cell lines tested.


De Novo Truncating Mutations in the Last and Penultimate Exons of PPM1D Cause an Intellectual Disability Syndrome.
Intellectual disability (ID) is a highly heterogeneous disorder involving at least 600 genes, yet a genetic diagnosis remains elusive in ∼35%-40% of individuals with moderate to severe ID. Recent meta-analyses statistically analyzing de novo mutations in >7,000 individuals with neurodevelopmental disorders highlighted mutations in PPM1D as a possible cause of ID. PPM1D is a type 2C phosphatase that functions as a negative regulator of cellular stress-response pathways by mediating a feedback loop of p38-p53 signaling, thereby contributing to growth inhibition and suppression of stress-induced apoptosis. We identified 14 individuals with mild to severe ID and/or developmental delay and de novo truncating PPM1D mutations. Additionally, deep phenotyping revealed overlapping behavioral problems (ASD, ADHD, and anxiety disorders), hypotonia, broad-based gait, facial dysmorphisms, and periods of fever and vomiting. PPM1D is expressed during fetal brain development and in the adult brain. All mutations were located in the last or penultimate exon, suggesting escape from nonsense-mediated mRNA decay. Both PPM1D expression analysis and cDNA sequencing in EBV LCLs of individuals support the presence of a stable truncated transcript, consistent with this hypothesis. Exposure of cells derived from individuals with PPM1D truncating mutations to ionizing radiation resulted in normal p53 activation, suggesting that p53 signaling is unaffected. However, a cell-growth disadvantage was observed, suggesting a possible effect on the stress-response pathway. Thus, we show that de novo truncating PPM1D mutations in the last and penultimate exons cause syndromic ID, which provides additional insight into the role of cell-cycle checkpoint genes in neurodevelopmental disorders.


Financial Toxicity.
to identify evidence in the literature about financial toxicity in cancer patients during chemotherapy treatment. this is a mini review with search in the following databases: Cumulative Index to Nursing and Allied Health Literature, Latin American and Caribbean Health Sciences Literature, National Library of Medicine and Scopus. Articles were surveyed from July 2018 to February 2019 using the following descriptors and Boolean operators: "financial toxicity" OR "financial difficulties" AND "cancer" AND "chemotherapy". the search totaled 64 articles. After exclusion of duplicates that did not meet the inclusion criteria, the final sample was composed of five articles published in 2013, 2015, 2017 and 2018. Financial difficulty caused by cancer treatment and its impact on quality of life was a frequent expression used in the studies. studies show that financial toxicity damages the quality of life of patients. A scarcity of studies on the subject was also observed, possibly because this is a new concept.


Effectiveness of Folic Acid Fortified Flour for Prevention of Neural Tube Defects in a High Risk Region.
Despite efforts to tackle folate deficiency and Neural Tube Defects (NTDs) through folic acid fortification, its implementation is still lacking where it is needed most, highlighting the need for studies that evaluate the effectiveness of folic acid fortified wheat flour in a poor, rural, high-risk, NTD region of China. One of the most affected regions, Shanxi Province, was selected as a case study. A community intervention was carried out in which 16,648 women of child-bearing age received fortified flour (eight villages) and a control group received ordinary flour (three villages). NTD birth prevalence and biological indicators were measured two years after program initiation at endline only. The effect on the NTD burden was calculated using the disability-adjusted life years (DALYs) method. In the intervention group, serum folate level was higher than in the control group. NTDs in the intervention group were 68.2% lower than in the control group (OR = 0.313, 95% CI = 0.207-0473, p < 0.001). In terms of DALYs, burden in intervention group was approximately 58.5% lower than in the control group. Flour fortification was associated with lower birth prevalence and burden of NTDs in economically developing regions with a high risk of NTDs. The positive findings confirm the potential of fortification when selecting an appropriate food vehicle and target region. As such, this study provides support for decision makers aiming for the implementation of (mandatory) folic acid fortification in China.


Struggling to return to everyday life-The experiences of quality of life 1 year after delirium in the intensive care unit.
Many critically ill patients report a change in their health-related quality of life after intensive care unit (ICU) discharge. Patients who experience delirium during their ICU stay are perceived as a fragile group of ICU survivors, and the 'quality of life' phenomenon needs to be studied among these patients. To explore everyday life experiences of critically ill patients with delirium during the ICU stay, from ICU discharge until 1-year follow-up, focusing on their health-related quality of life and cognitive function. We used a descriptive qualitative research design and interviewed patients 1 year after ICU admission. The participants were recruited from a pre-planned one-year follow-up study of 'Agents Intervening against Delirium for patients in the Intensive Care Unit trial'. Data were analysed using Framework Analysis Method and content analysis. Nine women and eight men participated and reported a struggle when returning to everyday life or adapting to a new normality from hospital discharge to 1 year later. None of the participants had been aware of the challenges they would face after hospital discharge. They described a need for more information about these challenges to themselves and about primary care to better understand their situation and the struggles they experience during recovery. One overall theme emerged from the analysis 'From enduring to adapting' with three subthemes: 'Struggling to regain a functional life', 'Struggling to regain normal cognition' and 'Distressing manifestations from the ICU'. To improve recovery and the quality of rehabilitation for critically ill patients suffering from delirium, it is essential to understand the phenomenon of ICU survivorship and what this fragile group of patients is going through. It is necessary to bridge the gap between secondary and primary care so patients can receive optimal training and support when needed. Bridging the gap between primary and secondary healthcare services is urgently needed to improve rehabilitation for ICU survivors after critical illness.


Uniqueness and estimation of three-dimensional motion parameters of rigid objects with curved surfaces.
Two main results are established in this paper. First, we show that seven point correspondences are sufficient to uniquely determine from two perspective views the three-dimensional motion parameters (within a scale factor for the translations) of a rigid object with curved surfaces. The seven points should not be traversed by two planes with one plane containing the origin, nor by a cone containing the origin. Second, a set of ``essential parameters'' are introduced which uniquely determine the motion parameters up to a scale factor for the translations, and can be estimated by solving a set of linear equations which are derived from the correspondences of eight image points. The actual motion parameters can subsequently be determined by computing the singular value decomposition (SVD) of a 3×3 matrix containing the essential parameters.


Antenatal Periviability Counseling and Decision Making: A Retrospective Examination by the Investigating Neonatal Decisions for Extremely Early Deliveries Study Group.
To describe periviability counseling practices and decision making. This is a retrospective review of mothers and newborns delivering between 22 and 24 completed weeks from 2011 to 2015 at six U.S. centers. Maternal and fetal/neonatal clinical and maternal sociodemographic data from medical records and geocoded sociodemographic information were collected. Separate analyses examined characteristics surrounding receiving neonatology consultation; planning neonatal resuscitation; and centers' planned resuscitation rates. Neonatology consultations were documented for 40, 63, and 72% of 498 mothers delivering at 22, 23, and 24 weeks, respectively. Consult versus no-consult mothers had longer median admission-to-delivery intervals (58.7 vs. 8.7 h, p < 0.001). Consultations were seen more frequently when parental decision making was evident. In total, 76% of mothers had neonatal resuscitation planned. Resuscitation versus no-resuscitation newborns had higher mean gestational ages (24.0 vs. 22.9 weeks, p < 0.001) and birthweights (618 vs. 469 g, p < 0.001). Planned resuscitation rates differed at higher (HR) versus lower (LR) rate centers at 22 (43 vs. 7%, p < 0.001) and 23 (85 vs. 58%, p < 0.001) weeks. HR versus LR centers' populations had more socioeconomic hardship markers but fewer social work consultations (odds ratio: 0.31; confidence interval: 0.15-0.59, p < 0.001). Areas requiring improvement included delivery/content of neonatology consultations, social work support, consideration of centers' patient populations, and opportunities for shared decisions.


Vasoactive prostanoids are generated from arachidonic acid by COX-1 and COX-2 in the mouse.
Generation of vasoactive prostanoids from arachidonic acid by cyclooxygenase (COX)-1 and COX-2 was investigated in anesthetized mice. Intravenous injections of the prostanoid precursor arachidonic acid increased pulmonary arterial pressure and decreased systemic arterial pressure. Pulmonary pressor and systemic depressor responses were attenuated by SC-560 and nimesulide, inhibitors of COX-1 and COX-2, in doses that did not alter responses to injected prostanoids. Pulmonary pressor responses to arachidonic acid were blocked and a depressor response was unmasked, whereas systemic depressor responses were not altered, by a thromboxane receptor antagonist. Pulmonary and systemic pressor responses to angiotensin II injections and systemic pressor responses to angiotensin II infusion were not modified by COX-1 or COX-2 inhibitors but were attenuated by losartan. Systemic depressor responses to arachidonic acid were smaller in COX-1 and COX-2 knockout mice, whereas responses to angiotensin II, norepinephrine, U-46619, endothelin-1, and PGE(1) were not different in COX-1 and COX-2 knockout and wild-type control mice. These results suggest that vasoactive prostanoids with pulmonary pressor and systemic vasodepressor activity are formed by COX-1 and COX-2 and are consistent with Western blot analysis and immunostaining showing the presence of COX-1 and COX-2. These data suggest that thromboxane A(2) (TxA(2)) is formed from the precursor by COX-1 and COX-2 in the lung and are in agreement with immunofluorescence studies showing thromboxane synthase. The present data suggest that COX-1- or COX-2-derived prostanoids do not modulate responses to angiotensin II or other vasoactive agents and that prostanoid responses are similar in CD-1 and C57BL/6 and in male and female mice.


Characterization of Drug-Related Problems Occurring in Patients Receiving Outpatient Antimicrobial Therapy.
Despite the numerous benefits of outpatient parenteral antimicrobial therapy (OPAT), appreciable risks of drug-related problems (DRPs) exist. No studies to date comprehensively assess DRPs in this population. Objectives of this study were to (1) characterize the frequency and types of DRPs experienced by patients discharged on OPAT and (2) determine the fraction of adverse drug reactions (ADRs) resulting in hospital readmission or emergency department (ED) presentation and changes in therapy. This was a retrospective chart analysis evaluating consecutive adult patients discharged on OPAT between May 2015 and October 2015. Patients were assessed for the presence of DRPs until the cessation of antimicrobial treatment, including oral step-down therapy. The outcome of each ADR was recorded, including those resulting in hospital readmissions, presentation to the ED, or changes in antimicrobials. Among 144 patients discharged on OPAT, 199 DRPs occurred in 91 (63.2%) patients. Harm and potential impaired efficacy occurred in 76.9% and 23.1%, respectively. The ADRs comprised 59% of DRPs, occurring in 44.4% of patients. The second most common DRP type was drug interactions (DIs), accounting for 22.6% of DRPs. Rifampin, fluoroquinolones, and daptomycin had the highest frequencies of preventable DRPs in the form of DIs, whereas cephalosporins had the fewest DRPs. Approximately 26% of ADRs caused changes in therapy and 9% resulted in hospital readmission or ED utilization. DRPs with the potential to cause patient harm or impair treatment efficacy often occur with OPAT, most commonly ADRs and DIs. Enhanced monitoring and transitions of care management may reduce the incidence of these DRPs.


HCFA tightens eligibility requirements for provider-based status.
On October 10, 2000, HCFA will implement new eligibility standards for all off-campus entities with a provider-based designation. Providers that wish to retain the provider-based status of their ambulatory care facilities need to take action to more fully integrate the ambulatory care facilities with their main facilities. In addition, the provider will need to show that 75 percent of the patient population served by the ambulatory care facility also is served by the provider's main facility.


Antitumor effect of matrine in human hepatoma G2 cells by inducing apoptosis and autophagy.
To study the antitumor effect of matrine in human hepatoma G2 (HepG2) cells and its molecular mechanism involved in antineoplastic activities. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to detect viability of HepG2 cells. The effect of matrine on cell cycle was detected by fl ow cytometry. Annexin-V-FITC/PI double staining assay was used to detect cellular apoptosis. Cellular morphological changes were observed under an inverted phase contrast microscope. Transmission electron microscopy was performed to further examine ultrastructural structure of the cells treated with matrine. Monodansylcadaverine (MDC) staining was used to detect autophagy. Whether autophagy is blocked by 3-methyladenine (3-MA), an autophagy inhibitor, was evaluated. Expression levels of Bax and Beclin 1 in HepG2 cells were measured by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR). Matrine significantly inhibited the proliferation of HepG2 cells in a dose- and time-dependent manner, and induced G1-phase cell cycle arrest and apoptosis of HepG2 cells in a dose-dependent manner. The total apoptosis rate was 0.14% for HepG2 cells not treated with matrine. In contrast, the apoptosis rate was 28.91%, 34.36% and 38.80%, respectively, for HepG2 cells treated with matrine at the concentration of 0.5, 1.0 and 2.0 mg/mL. The remarkable morphological changes were observed under an inverted phase contrast microscope. Abundant cytoplasmic vacuoles with varying sizes were observed in HepG2 cells treated with matrine. Furthermore, vacuolization in cytoplasm progressively became larger and denser when the concentration of matrine was increased. Electron microscopy demonstrated formation of abundant autophagic vacuoles in HepG2 cells after matrine treatment. When the specific autophagic inhibitor, 3-MA, was applied, the number of autophagic vacuoles greatly decreased. MDC staining showed that the fluorescent density was higher and the number of MDC-labeled particles in HepG2 cells was greater in matrine treatment group than in control group. Fewer autophagic vacuoles were observed in the combined 3-MA and matrine treatment group when 3-MA was added before matrine treatment, indicating that both autophagy and apoptosis are activated when matrine-induced death of hepatoma G2 cells occurs. Real-time quantitative RT-PCR revealed that the expression levels of Bax gene, an apoptosis-related molecule, and Beclin 1 gene which plays a key role in autophagy were higher in matrine treatment group than in control group, indicating that Beclin 1 is involved in matrine-induced autophagy and the pro-apoptotic mechanism of matrine may be related to its upregulation of Bax expression. Matrine has potent antitumor activities in HepG2 cells and may be used as a novel effective reagent in treatment of hepatocellular carcinoma.


Steroid induction of therapy-resistant cytokeratin-5-positive cells in estrogen receptor-positive breast cancer through a BCL6-dependent mechanism.
Therapy resistance remains a major problem in estrogen receptor-α (ERα)-positive breast cancer. A subgroup of ERα-positive breast cancer is characterized by mosaic presence of a minor population of ERα-negative cancer cells expressing the basal cytokeratin-5 (CK5). These CK5-positive cells are therapy resistant and have increased tumor-initiating potential. Although a series of reports document induction of the CK5-positive cells by progestins, it is unknown if other 3-ketosteroids share this ability. We now report that glucocorticoids and mineralocorticoids effectively expand the CK5-positive cell population. CK5-positive cells induced by 3-ketosteroids lacked ERα and progesterone receptors, expressed stem cell marker, CD44, and displayed increased clonogenicity in soft agar and broad drug-resistance in vitro and in vivo. Upregulation of CK5-positive cells by 3-ketosteroids required induction of the transcriptional repressor BCL6 based on suppression of BCL6 by two independent BCL6 small hairpin RNAs or by prolactin. Prolactin also suppressed 3-ketosteroid induction of CK5+ cells in T47D xenografts in vivo. Survival analysis with recursive partitioning in node-negative ERα-positive breast cancer using quantitative CK5 and BCL6 mRNA or protein expression data identified patients at high or low risk for tumor recurrence in two independent patient cohorts. The data provide a mechanism by which common pathophysiological or pharmacologic elevations in glucocorticoids or other 3-ketosteroids may adversely affect patients with mixed ERα+/CK5+ breast cancer. The observations further suggest a cooperative diagnostic utility of CK5 and BCL6 expression levels and justify exploring efficacy of inhibitors of BCL6 and 3-ketosteroid receptors for a subset of ERα-positive breast cancers.


Millimeter-wave absorption by cutaneous blood vessels: a computational study.
The aims of the present study were to calculate the specific absorption rate (SAR) and E-field distributions inside cutaneous blood vessels and in surrounding tissues (dermis and fat) depending on the frequency of millimeter wave exposure. Most calculations were performed using the finite-difference time-domain (FDTD) technique. A rectangular block of homogeneous or multilayer tissue with blood vessels located in the center of the block was used as the basic geometry. We found that the SAR reached its maximal value in a long blood vessel oriented parallel to the E-field. It exceeded the SAR in the surrounding dermis by 40%-42% at 42.25 GHz. However, in the same blood vessel oriented perpendicularly to the E-field, the SAR was lower than that of the surrounding dermis. Absorption of millimeter waves in a cutaneous blood vessel was higher at 61.22 GHz than at 42.25 GHz. The SAR distribution in a blood vessel was nearly uniform. Because of the small sizes of cutaneous blood vessels relative to the wavelength, the SAR distributions in these blood vessels can be calculated by using quasi-static theory.


Dominant collagen VI mutations are a common cause of Ullrich congenital muscular dystrophy.
Mutations in the three collagen VI genes COL6A1, COL6A2 and COL6A3 cause Bethlem myopathy and Ullrich congenital muscular dystrophy (UCMD). UCMD, a severe disorder characterized by congenital muscle weakness, proximal joint contractures and marked distal joint hyperextensibility, has been considered a recessive condition, and homozygous or compound heterozygous mutations have been defined in COL6A2 and COL6A3. In contrast, the milder disorder Bethlem myopathy shows clear dominant inheritance and is caused by heterozygous mutations in COL6A1, COL6A2 and COL6A3. This model, where dominant mutations cause mild Bethlem myopathy and recessive mutations cause severe UCMD was recently challenged when a patient with UCMD was shown to have a heterozygous in-frame deletion in COL6A1. We have studied five patients with a clinical diagnosis of UCMD. Three patients had heterozygous in-frame deletions in the N-terminal region of the triple helical domain, one in the alpha1(VI) chain, one in alpha2(VI) and one in alpha3(VI). Collagen VI protein biosynthesis and assembly studies showed that these mutations act in a dominant negative fashion and result in severe collagen VI matrix deficiencies. One patient had recessive amino acid changes in the C2 subdomain of alpha2(VI), which prevented collagen VI assembly. No collagen VI mutations were found in the fifth patient. These data demonstrate that rather than being a rare cause of UCMD, dominant mutations are common in UCMD, now accounting for four of the 14 published cases. Mutation detection in this disorder remains critical for accurate genetic counseling of patients and their families.


Antibody Attributes that Predict the Neutralization and Effector Function of Polyclonal Responses to SARS-CoV-2.
While antibodies provide significant protection from SARS-CoV-2 infection and disease sequelae, the specific attributes of the humoral response that contribute to immunity are incompletely defined. In this study, we employ machine learning to relate characteristics of the polyclonal antibody response raised by natural infection to diverse antibody effector functions and neutralization potency with the goal of generating both accurate predictions of each activity based on antibody response profiles as well as insights into antibody mechanisms of action. To this end, antibody-mediated phagocytosis, cytotoxicity, complement deposition, and neutralization were accurately predicted from biophysical antibody profiles in both discovery and validation cohorts. These predictive models identified SARS-CoV-2-specific IgM as a key predictor of neutralization activity whose mechanistic relevance was supported experimentally by depletion. Validated models of how different aspects of the humoral response relate to antiviral antibody activities suggest desirable attributes to recapitulate by vaccination or other antibody-based interventions.


Melanonychia striata longitudinalis. A case report.
Melanonychia strata longitudinalis is a pigmented (dark brown-tan-black) band running in the long axis of the nail. This abnormal color of the nail plate is caused by a focal increase of melanin because of increased number and/or function of normal or abnormal melanocytes in the nail matrix. Pigmented streaks in nails due to melanocytic hyperplasia are said to be so exceedingly common in black persons as to be normal and the phenomenon is benign in the majority of cases. In our white patient reported here the pigmentation is due to epithelial hyperpigmentation without melanocytic hyperplasia or evidence of malignancy, hormonal abnormality, or other adventitious influence.


Paracrine-mediated neuroprotection and neuritogenesis of axotomised retinal ganglion cells by human dental pulp stem cells: comparison with human bone marrow and adipose-derived mesenchymal stem cells.
We have investigated and compared the neurotrophic activity of human dental pulp stem cells (hDPSC), human bone marrow-derived mesenchymal stem cells (hBMSC) and human adipose-derived stem cells (hAMSC) on axotomised adult rat retinal ganglion cells (RGC) in vitro in order to evaluate their therapeutic potential for neurodegenerative conditions of RGC. Using the transwell system, RGC survival and length/number of neurites were quantified in coculture with stem cells in the presence or absence of specific Fc-receptor inhibitors to determine the role of NGF, BDNF, NT-3, VEGF, GDNF, PDGF-AA and PDGF-AB/BB in stem cell-mediated RGC neuroprotection and neuritogenesis. Conditioned media, collected from cultured hDPSC/hBMSC/hAMSC, were assayed for the secreted growth factors detailed above using ELISA. PCR array determined the hDPSC, hBMSC and hAMSC expression of genes encoding 84 growth factors and receptors. The results demonstrated that hDPSC promoted significantly more neuroprotection and neuritogenesis of axotomised RGC than either hBMSC or hAMSC, an effect that was neutralized after the addition of specific Fc-receptor inhibitors. hDPSC secreted greater levels of various growth factors including NGF, BDNF and VEGF compared with hBMSC/hAMSC. The PCR array confirmed these findings and identified VGF as a novel potentially therapeutic hDPSC-derived neurotrophic factor (NTF) with significant RGC neuroprotective properties after coculture with axotomised RGC. In conclusion, hDPSC promoted significant multi-factorial paracrine-mediated RGC survival and neurite outgrowth and may be considered a potent and advantageous cell therapy for retinal nerve repair.


Development of an enzyme-linked immunosorbent assay system to determine the presence of antibodies specific for taxane structures.
Taxanes, which are widely used in treatment of numerous cancer types, are well-known to induce hypersensitivity reactions (HSR), especially in the case of paclitaxel. Although the cause of the HSR is commonly thought to be a non-immunological direct effect of the diluent which is used to dissolve paclitaxel, some reports suggest the possibility of the presence of an immunological reaction to the common taxane structure. The aim of this study was to establish a method to determine the presence of anti-taxane antibodies in body fluids of patients who have previously received paclitaxel, in order to estimate the risk of the occurrence of HSR to other taxane compounds, such as docetaxel. To prepare an enzyme-linked immunosorbent assay (ELISA) plate for determining taxanes, 10-deacetylbaccatin III (DAB) was first succinylated by use of dimethylaminopyridine and succinic anhydride in dried pyridine. After the succinylation reaction, three different products were obtained, and these were confirmed as 7-succinoyl DAB (7-DAB), 10-succinoyl DAB (10-DAB), and 7,10-disuccinoyl DAB (7,10-DAB) by (1)H-NMR analysis. Each of these three products was conjugated with bovine serum albumin (BSA), and adsorbed on an ELISA plate. By using a commercially available anti-taxane monoclonal antibody as a model antibody, the detection limit of the anti-taxane antibodies on the 7-DAB-BSA-, 10-DAB-BSA-, and 7,10-DAB-BSA-conjugated ELISA plate was estimated as 0.3, 1 and 10 ng/ml, respectively. The ELISA system established in this study may therefore be useful for estimating the risk of HSR to taxanes in a patient prior to the use of these drugs.


Genetic evaluation of oocyte donors: recipient couple preferences and outcome of testing.
To report preferences of recipient couples for genetic testing of their oocyte donors. Observational report of results from a genetic testing-options form implemented as part of routine care. Private practice. Recipients and oocyte donors. Recipient couples completed a form before screening of their oocyte donor that outlined required screening and recommended tests that the couple could accept or decline. Couples were given information about carrier frequency, risk to their child if results were abnormal, and cost. Percentage of couples accepting optional testing. Of the 63 couples with data available from their first testing-options form, 42 (67%) accepted and 21 (33%) declined fragile X testing, whereas 34 (54%) accepted and 29 (46%) declined karyotyping. When asked whether they would accept additional testing of their donor if it was recommended by a genetic counselor, 15 (24%) said that they would accept additional testing regardless of cost, 35 (56%) declined, and 13 (20%) indicated that their decision would depend on the cost. In many cases, history was elicited by the genetic counselor or test results were obtained that influenced further testing, decisions to proceed, or provided information important for the child. Recipient couples sometimes chose to decline tests that we recommended but did not require, despite the relatively low cost of this testing compared with the total cost of the oocyte donation cycle.


Financing health care for disabled children.
Information about health care use, charges, and out-of-pocket expenses is critical to the development of an equitable and efficient treatment system for disabled children. Data from the 1980 National Medical Care Utilization and Expenditure Survey were used, and differences in use, charges, and out-of-pocket expenses for children with and without limitations in their activities due to chronic health problems are described. The results indicate that children limited in their activities used more medical services than other children, especially hospital-based services and services provided by health professionals other than physicians. Charges and out-of-pocket expenses were two to three times higher on average for disabled children, compared with other children. Charges and out-of-pocket expenses were also skewed; 10% of the sample children accounted for more than 60% of total charges and out-of-pocket expenses for the disabled population. The skewed distribution of out-of-pocket expenses suggests that financial burdens are unevenly shared by families of disabled children. Several public policy options designed to result in a more equitable distribution of financial risks are discussed.


Primary lymphoma of the liver - a case-report and a review of the literature.
A case of primary lymphoma of the liver in a 72-year-old man is reported. The patient showed a three to four month history of abdominal ache, low grade fever, and weight loss. The presence of a large mass occupying the V and VT hepatic segments was demonstrated; no other organs were involved and no evidence of lymphoadenopathy was found. Laboratory tests showed a remarkable increase in ESR, LDH and alkaline phosphatase while CEA and alpha-1 fetoprotein were within the normal values. Two ultrasound-guided needle biopsies on the hepatic lesion did not produce any significant tissue for diagnosis. Therefore the V and VI hepatic segments were excised; a hard, greyish mass with the largest diameter of 6.5 cm, was found at laparatomy. Histology of the surgical specimen showed the presence of a malignant non-Hodgking's B-cell lymphoma, centroblastic-centrocytic, diffuse type (working formulation: G) of intermediate grade of malignancy. Staging was completed with lymphography, and bone marrow biopsy; both proved normal. After the operation adjuvant chemotherapy was commenced (COP for six cycles). Follow-up has so far shown no signs of recurrence after 24 months. In conclusion, the diagnosis of primary lymphoma of the liver is very difficult and is often reached only after histologic evaluation of the surgical specimen. Moreover, this disease has a much more favourable prognosis than the hepatocellular carcinoma and other primary or secondary liver cancers; therefore accurate investigations are important to reach the correct diagnosis.


Zoster vaccine (Zostavax): a review of its use in preventing herpes zoster and postherpetic neuralgia in older adults.
Individuals who have been infected with varicella zoster virus (VZV) are at risk for developing herpes zoster and this risk appears to be related to a decline in VZV-specific cell-mediated immunity (CMI). Zostavax (zoster vaccine) is a one-dose, high-potency, live, attenuated VZV vaccine that boosts VZV-specific CMI and this is its presumed mechanism of action. Zoster vaccine is registered in the EU for use in adults aged >or=50 years for the prevention of herpes zoster and herpes zoster-related postherpetic neuralgia. In the Shingles Prevention Study, a placebo-controlled trial in adults aged >or=60 years (n = 38 546), zoster vaccine led to a sustained boost of VZV-specific CMI. Over a mean herpes zoster surveillance period of 3.1 years, zoster vaccine reduced the herpes zoster-related burden of illness by 61%, reduced the incidence of herpes zoster by 51% and reduced the incidence of postherpetic neuralgia by 67%. Zoster vaccine recipients who developed herpes zoster had a shorter illness duration and severity than placebo recipients who developed herpes zoster. Zoster vaccine had continuing efficacy in a Shingles Prevention Study subpopulation followed for 7 years post-vaccination. Zoster vaccine was generally well tolerated in older adults. While cost-effectiveness estimates in pharmacoeconomic analyses varied widely according to vaccine and herpes zoster parameter cost/benefit estimates, an analysis from a UK perspective found a zoster vaccine immunization programme in adults aged 65 years to be cost effective. In older adults, the zoster vaccine has the potential to significantly reduce the herpes zoster burden of illness by decreasing the incidence of herpes zoster or reducing its severity.


[Eikenella corrodens: five year experience in a general hospital].
Eikenella corrodens is a Gram negative bacillus that colonizes oropharyngeal and digestive mucous membranes. It is difficult to isolate in the laboratory. This might be the reason why its implication in different types of infection is not well known. We report a five year experience in a general hospital. Charts of 125 infections have been retrospectively reviewed. No selective medium was employed for the isolation. Infections were related to oral cavity in 68 patients (54%) and to gastrointestinal tract in 45 (36%); in 12 no clear source could be found. The prevailing infections were: postsurgical 32, apendiceal 22, abscesses 22, and traumatic wounds 12. Seventy two percent of the patients were adults and 93% were hospitalized. Eighteen patients had a neoplasia, 6 were diabetic, and 3 had some kind of immunodeficiency. Etiology was polymicrobial in 91% of the cases, the prevailing association being with Streptococcus spp. (74%). Only 15 patients had a pure culture, 11 of which had received previous antibiotic treatment. The most serious infections were a subdural empyema secondary to sinusitis, a mediastinitis following a mouth floor abscess, and a peritonitis with pleural empyema preceded by an hepatic abscess. The three were polymicrobial. Outcome was always satisfactory with no associated death. The origin of these infections is almost always clearly related with its habitat, but their spectrum is wide. Polymicrobial condition of most of them makes diagnosis as well as interpretation of its clinical meaning difficult. Nevertheless, association of E. corrodens with serious or slow healing infections fully justifies the efforts towards its isolation and the choice of antibiotics to which it is susceptible.


MONICA quality assessments.
The authors give an overview of the quality assessments in the WHO MONICA project and compare the quality of the data from Northern Sweden with other reporting units. Standardized measurement procedures and routine checks were used to ensure good quality of the data. The quality has been evaluated by a scoring system. The results show that the data from Northern Sweden have good quality for all variables except total cholesterol in the initial survey. In the subsequent surveys, the quality of cholesterol data was good. Great effort was put in to ensure good data quality and Northern Sweden is one of the MONICA units with very good quality of data.


Pharmacists' experiences with dispensing opioids: provincial survey.
To explore pharmacists' beliefs, practices, and experiences regarding opioid dispensing. Mailed survey. The province of Ontario. A total of 1011 pharmacists selected from the Ontario College of Pharmacists' registration list. Pharmacists' experiences with opioid-related adverse events (intoxication and aberrant drug-related behaviour) and their interactions with physicians. A total of 652 pharmacists returned the survey, for a response rate of 64%. Most (86%) reported that they were concerned about several or many of their patients who were taking opioids; 36% reported that at least 1 patient was intoxicated from opioids while visiting their pharmacies within the past year. Reasons for opioid intoxication included the patient taking more than prescribed (84%), the patient using alcohol or sedating drugs along with the opioid (69.9%), or the prescribed dose being too high (34%). Participants' most common concerns in the 3 months before the survey were patients coming in early for prescription refills, suspected double-doctoring, and requests for replacement doses for lost medication (reported frequently by 39%, 12%, and 16% of respondents, respectively). Pharmacists were concerned about physician practices, such as prescribing benzodiazepines along with opioids. Pharmacists reported difficulty in reaching physicians directly by telephone (43%), and indicated that physicians frequently did not return their calls promptly (28%). The strategies rated as most helpful for improving opioid dispensing were a provincial prescription database and opioid prescribing guidelines. Pharmacists commonly observe opioid intoxication and aberrant drug-related behaviour in their patients but have difficulty communicating their concerns to physicians. System-wide strategies are urgently needed to improve the safety of opioid prescribing and to enhance communication between physicians and pharmacists.


Factors associated with racial/ethnic differences in colorectal cancer screening.
Racial/ethnic differences in colorectal cancer (CRC) screening rates are thought to account, in part, for the racial/ethnic differences in CRC disease burden. The purpose of this study was to examine which factors mediate racial/ethnic differences in CRC screening. Five hundred sixty participants attending a primary care clinic, aged 50 to 80 years, and of African-American, Hispanic, or non-Hispanic white race/ethnicity were interviewed. The goal was to assess the contribution of sociodemographic characteristics, knowledge, beliefs about CRC, and the health care experience with their primary care doctor to racial/ethnic differences in CRC screening. The outcome variable was self-reported screening. All analyses were weighted; bivariate testing and multivariate logistic regression was conducted. The response rate was 55.7%, with no sociodemographic differences noted between respondents and non-respondents. Respondents were African-American (n = 194), Hispanic (n = 162), and non-Hispanic white (n = 204); 64.5% were aged 50 to 64 years; 63.1% were women; 96.9% were insured; and over half reported a total annual income of less than $25,000. Overall 62.5% were current with CRC screening: 67.5% of non-Hispanic whites, 54.3% of African-Americans, and 48.6% of Hispanics (P < .001). A doctor's recommendation (odds ratio, 3.86); awareness of screening (odds ratio, 3.32); older age (odds ratio, 2.88); greater education (odds ratio, 2.02); and perceived susceptibility (odds ratio, 1.74) contributed to racial/ethnic differences in CRC screening. Interventions to address CRC screening disparities among racial/ethnic groups should focus on the health care setting and patient education about CRC screening; differences in attitudes and beliefs seem to be less important.


Filamentation in Atmospheric Air with Tunable 1100-2400 nm Near-Infrared Femtosecond Laser Source.
Intense femtosecond pulse filamentation in open-air has been utilized for long distance optical communication and remote sensing, but it results in nonlinear-effect driven eye hazards which are not addressed by current eye safety standards. A systematic study of filamentation in atmospheric air was performed using a tunable 100 fs near-infrared laser (1100 nm-2400 nm). While undergoing filamentation, each source wavelength was spectrally broadened resulting in supercontinuum and third harmonic generation in the visible and near-IR spectrum. We record the spectra at the center and fringes of the supercontinuum as it is imaged onto a planar surface. In a full beam collection regime, we report the energy of the sub-1000 nm light generation for source wavelengths from 1100 nm to 1600 nm and compare the energy density to the maximum permissible exposure values under the ANSI Z136.1 laser safety standard.


Enabling scale-up of mesoporous silicon for lithium-ion batteries: a systematic study of a thermal moderator.
The volume expansion of silicon during cycling of a lithium-ion battery (LIB) leads to degradation and capacity loss. Porous silicon can address many of the issues faced by silicon active materials and has previously been shown to have excellent cyclability. Recently we have uncovered the mechanisms underpinning the pore evolution in magnesiothermic reduction (MgTR) of silica and further demonstrated that it has the potential to produce porous silicon in a scalable and economic manner [J. Mater. Chem. A, 2020, 8, 4938]. However, the scalability of MgTR is affected by the large excess heat produced during reaction. Although previous studies have shown that NaCl can be used as a thermal moderator to mitigate this issue, this has not been systematically investigated, leading to a lack of knowledge on scalability of MgTR. Here, by carefully investigating the roles of NaCl, we show that the NaCl ratio and reduction temperature are the critical factors for controlling scale-up and the product properties. We identified the upper temperature limit of NaCl as a thermal moderator. Further, we systematically showed how the amount of NaCl and the reduction temperature affect the porous properties of the product silicon. Our results have established pathways for scaling-up this method such that it can now be taken forward to target specific porous silicon properties.


The thermoelastic properties of monolayer covalent organic frameworks studied by machine-learning molecular dynamics.
Two-dimensional (2D) covalent organic frameworks (COFs) are emerging as promising 2D polymeric materials with broad applications owing to their unique properties, among which the mechanical properties are quite important for various applications. However, the mechanical properties of 2D COFs have not been systematically studied yet. Herein, a machine-learned neuroevolution potential (NEP) was developed to study the elastic properties of two representative monolayer 2D COFs, namely COF-1 and COF-5. The trained NEP enables one to study the elastic properties of 2D COFs in realistic situations (e.g., finite size and temperature) and possesses greatly improved computational efficiency when compared with density functional theory calculations. With the aid of the obtained NEP, molecular dynamics (MD) simulations together with a strain-fluctuation method were employed to evaluate the elastic constants of the considered 2D COFs at different temperatures. The elastic constants of COF-1 and COF-5 monolayers were found to decrease with an increase in the temperature, though they were almost isotropic irrespective of the temperature. The thermally induced softening of 2D COFs below a critical temperature was observed, which is mainly attributed to their inherent ripple configurations at finite temperatures, while above the critical temperature, the damping effect of anharmonic vibrations became the dominant factor. Based on the proposed mechanisms, analytical models were developed for capturing the temperature dependence of elastic constants, which were found to agree with the MD simulation results well. This work provides an in-depth insight into the thermoelastic properties of monolayer COFs, which can guide the development of 2D COF materials with tailored mechanical behaviors for enhancing their performance in various applications.


Infill Strategy in 3D Printed PLA Carbon Composites: Effect on Tensile Performance.
Tuning the infill pattern is one of the key features in additive manufacturing to optimise part weight. In this work, the effect of the infill strategy, including rate and pattern type, is studied on the mechanical performance of polylactic acid (PLA)-carbon composite. In particular, three types of patterns and four filling levels are combined. These combinations are evaluated by tensile loading applied on dogbone specimens. In addition, the underlined deformation mechanisms are further explored using filament-based finite element model. The numerical simulation is built from sliced models and converted into 3D meshes to predict tensile performance. The results show that the infill rate has a nonlinear effect on the density of PLA-carbon composites, and its magnitude depends on the complexity of the generated pattern. In addition, tensile loading is found to activate varied modes of shearing and uniaxial deformations depending on the pattern type. This leads to different profiles and rankings of the tensile performance and allows the infill strategy to significantly affect the part performance, along with its density.


Autophagy, Apoptosis, and Cell Proliferation in Exstrophy-Epispadias Complex.
To investigate the state of autophagy and its interactions with apoptosis and cell proliferation in patients who underwent successful early closure or delayed closure of exstrophy. They compared those outcomes with cell culture samples from patients with vesicoureteral reflux as control. Primary cultures of bladder smooth muscle cells (SMCs) were established from patients with successful neonatal bladder closure (group 1, N = 5), delayed closure because of small bladder template (group 2, N = 5), and vesicoureteral reflux as control (group 3, N = 5). The myogenicity of the cultures was determined using anti-Desmin antibody. Immunostainings for LC3 to assess autophagy and Ki67 to assess cell proliferation were applied. Apoptosis was assessed by the terminal deoxynucleotidyl transferase-mediated dUTP digoxigenin nick-end labeling assay. Autophagy marker (LC3) expression was significantly higher in the delayed closure group than in the other groups, whereas no significant difference was noted between the neonatal closure and the control groups. Apoptotic indices of the SMCs were remarkably higher in SMC cultures from the delayed closures than in the neonatal closure and the control groups. A significantly lower expression of proliferation marker (Ki67) in the delayed closure group compared with the control and the neonatal closure group was also of note. Patients with small bladder template and delayed closure showed upregulated autophagic process and increased apoptotic indices while experiencing a dramatic decrease in the proliferation of their bladder SMCs. Finally, the concept of manipulating autophagy may lead to promising outcomes for patients with bladder exstrophy in the future.


Disturbance induced dynamics of a tritrophic novel ecosystem.
Novel ecosystems formed by invasive plants provide a good opportunity to get insight into early dynamics and pattern formation of these ecosystems. The invasive black locust as host plant, Bruchophagus robiniae as host-specific seed predator and its parasitoids were the components of the studied tritrophic system. To investigate disturbance-driven dynamics of this system we created seed-vacated host plant patches in a field experiment. We removed all pods from selected patches of black locust resulting in an induced local extinction of seed predators and their parasitoids. We hypothesized that disturbance enhances top-down control by parasitoids; this enhanced top-down control decreases seed predation, facilitating the host plant's spread. We found that disturbance modified only parasitism after controlling with year effect: in vacated patches median parasitism was higher than in control patches. Seed predation exceeded its initial level in vacated patches in the third year after the disturbance, but in the fourth year it dropped again presumably due to the strong top-down control. Our findings also suggested that the seed predator was also affected by the bottom-up control of its host plant's density. We found that in the studied new ecosystem the top-down control was strengthened by the disturbance. Since the host plant of the tritrophic system is an invasive species, partial habitat disturbance of such species may increase the severity of parasitoid top-down control, which may reduce seed predation by the herbivores.


[Treatment of acute pancreatitis with 5-fluorouracil].
The authors developed an outline for the treatment of acute pancreatitis with 5-fluorouracil which consists in a single or double injection of 250 mg of the drug. The method was used in 54 acute pancreatitis cases. A single intravenous injection of 250 mg of the drug proved to be sufficient for arresting the acute process in edematous and edemo-hemorrhagic pancreatitis. This method helps to shorten the treatment period. Clinical data were confirmed by experimental findings in 12 dogs.


Roles of cytomegalovirus and Chlamydia trachomatis in the induction of cervical neoplasia in the mouse.
Cytomegalovirus and Chlamydia trachomatis are prevalent sexually transmissible pathogens. They produce persistent infections of the cervix and have been associated with cervical neoplasia. Cytomegalovirus has also been shown to induce transformation of cells in culture. Because of the high prevalence of genital infections with these pathogens and evidence that they may have oncogenic effects on the cervix, cytomegalovirus (strain AD-169) and C. trachomatis (serovar LGV-2) were tested for oncogenicity in a mouse model in which induction of cervical neoplasia by repeated exposure to inactivated herpes simplex viruses has been demonstrated previously. Cotton tampons, saturated with UV-inactivated cytomegalovirus, C. trachomatis, or corresponding control fluids, were inserted into the vaginas of virgin C57 mice 3 times a week. Smears of vaginal aspirates for cytological examination were obtained every 5 weeks. After 75-90 weeks of exposure, the mice were sacrificed and serial sections of their reproductive tracts were examined. Cervical dysplasia was detected by histological examination in 51% and cervical carcinoma in 10% of mice exposed to cytomegalovirus. In control mice, in contrast, dysplasia developed in 3% and carcinoma in none. The progression from normal cervical epithelium to dysplasia to carcinoma observed with cytomegalovirus exposure was similar to that observed previously in this model after exposure of mice to herpes simplex virus types 1 and 2. The frequencies of cervical abnormalities in mice exposed to C. trachomatis or corresponding control fluid were low, and differences between the two groups were not statistically significant. These data indicate that strain AD-169 of cytomegalovirus is oncogenic for the mouse cervix and suggest that the LGV-2 serovar of C. trachomatis is not.


BK virus as a mediator of graft dysfunction following kidney transplantation.
BK virus is a significant risk factor for kidney allograft dysfunction and loss among renal transplant recipients. Currently, there is no proven effective treatment except for the reduction of immunosuppression. In this review, we discuss diagnostic challenges and current treatment options for BK in kidney transplant recipients. Antiviral and antibiotic therapies have been employed for BK viraemia with variable efficacy. In addition, novel therapeutic regimens such as adoptive transfer of targeted T cells have been described as possible treatment options for recipients with BK nephropathy. BK can also be seen in the native kidneys of pancreas, heart, lung and liver transplant recipients, suggesting that BK screening measures should be employed to other solid organ transplant recipients. Early screening for BK combined with reduction of immunosuppression remains the mainstay of treatment for BK viraemia. New therapeutic advances demonstrate promise in vitro; however, the in-vivo efficacy will be demonstrated by future studies.


Indices for detecting differences in species composition: some simplifications of RDA and CCA.
We provide algebraic simplifications for the redundancy analysis (RDA) eigenvalue and the canonical correspondence analysis (CCA) eigenvalue in the special case of permanent plots sampled twice. The indices for RDA and CCA are interrelated and are intuitively interpretable. These simplifications also apply to simple split-plot designs and to a balanced design with two independent samples.


A critical analysis of Peru's HIV grant proposals to the Global Fund.
Peru has applied to six of the Global Fund to Fight AIDS, Tuberculosis and Malaria (the Global Fund) rounds for funding, achieving success on four occasions. The process of proposal development has, however, been criticised, especially concerning the use of evidence, relevance/consistency and performance indicators. We aimed to analyse the Peruvian Global Fund proposals according to those dimensions, providing feedback to improve future local efforts and inform global discussions around Global Fund procedures. We analysed the content of four HIV-focused proposals (rounds 2, 5, 6 and 8) regarding epidemic context, needs identification and prioritisation and monitoring and evaluation systems. Peruvian proposals submitted after round 1 were described as resulting from collaborative inputs involving formerly unrepresented sectors, principally 'vulnerable populations'. However, difficulties arose regarding the amount and quality of evidence about the epidemiological context; limited consideration of social determinants of the epidemic; lack of theory-driven interventions, and little synergy across projects and the inclusion of weak monitoring and evaluation systems, with poor indicators and measurement procedures. Prioritising the development of analytical and technical skills to generate Global Fund proposals would enhance the country's capacity to produce and utilise evidence, improve the technical-political interface, strengthen information systems and lead to more informed decision making and accountability.


Selectivity of 1-phenylimidazole as a ligand for cytochrome P-450 and as an inhibitor of microsomal oxidation.
Equilibrium dialysis studies established that 1-[4'-(3H)-phenyl]imidazole (PI) was bound to hepatic microsomal suspensions from control, phenobarbital (PB)- and 3-methylcholanthrene (3MC)-treated rats and that the binding was directly related to the cytochrome P-450 content. Computer-assisted Scatchard plot analysis of the binding data indicated the existence of two major types of microsomal binding sites in both control and induced rats, one with a high affinity (Ka approximately 1.5 X 10(7) M-1) and the other with a low affinity (Ka approximately 5 X 10(5) M-1) for PI. The binding of PI to the highly purified, individual cytochrome P-450s that constituted the major forms from the PB- and beta-naphthoflavone (beta NF)-induced rats exhibited affinities similar to the high and low affinity binding sites observed in microsomal suspensions. The two types of PI binding sites were characteristic of two classes of cytochrome P-450, and the major cytochrome induced by PB and 3MC (or beta NF) were each associated with one of these two classes. In concurrence with this, it was shown that, although PI was an excellent inhibitor of aromatic hydrocarbon hydroxylase (AHH) activity in PB-induced rats, it exhibited little or no inhibitory activity towards AHH activity in 3MC-induced animals.


Parathyroid hormone stimulates ammoniagenesis in canine renal proximal tubular segments.
To determine whether parathyroid hormone (PTH) affects ammoniagenesis in renal proximal tubule, we measured ammonia production in suspensions of canine renal proximal tubular segments incubated with 10 mM L-glutamine in the absence or presence of 10(-7) M PTH. Productions of ammonia were linear functions of time for 120 min and averaged 231 +/- 55 and 311 +/- 67 mumol ammonia/g protein in the absence and presence of PTH, respectively. When measured over the range of 10(-11)-10(-7) M PTH, half-maximal stimulation of ammonia production occurred between 10(-10) and 10(-9) M PTH. Maximal production of ammonia was observed at 10(-8) M PTH. Incubation of proximal tubular segments with PTH increased levels of adenosine 3',5'-cyclic monophosphate (cAMP) in vitro. Ammonia production was significantly enhanced by incubation of segments with the cAMP analogue, 8-bromoadenosine 3',5'-cyclic monophosphate. PTH also increased ammonia production in segments incubated with 1 mM L-glutamine. We conclude that PTH stimulates ammonia production in canine renal proximal tubular segments. This effect appears to be mediated, at least in part, through cAMP in vitro. Such stimulation could reflect a direct action of PTH on the proximal tubule to enhance ammoniagenesis in vivo.


[Relations between grandparents and grandchildren].
In spite of the steadily increasing number of grandparents there is only few research on this subject. In four studies we tried to elaborate forms of interaction between grandparents and grandchildren of different age groups. The samples comprised grandparent-grandchildren pairs, which were explored separately in an extensive way (altogether 232 subjects). Comparative results on contact-frequency, styles of grandparenting, functions of grandparents, meaning of the parent-generation and--only for the elder people--perception of own grandparents as well as satisfaction with the grandparent role are reported and discussed.


Simple HPLC-UV Method for Therapeutic Drug Monitoring of 12 Antiepileptic Drugs and Their Main Metabolites in Human Plasma.
The objective of the present report was to develop and validate a simple, selective, and reproducible high-performance liquid chromatography method with UV detection suitable for routine therapeutic drug monitoring of the most commonly used antiepileptic drugs and some of their metabolites. Simple precipitation of plasma proteins with acetonitrile was used for sample preparation. 10,11-dihydrocarbamazepine was used as an internal standard. Chromatographic separation of the analytes was achieved by gradient elution on a Phenyl-Hexyl column at 40 °C, using methanol and potassium phosphate buffer (25 mM; pH 5.1) as a mobile phase. The method was validated according to the FDA guidelines for bioanalytical method validation. It showed to be selective, accurate, precise, and linear over the concentration ranges of 1-50 mg/L for phenobarbital, phenytoin, levetiracetam, rufinamide, zonisamide, and lacosamide; 0.5-50 mg/L for lamotrigine, primidone, carbamazepine and 10-monohydroxycarbazepine; 0.2-10 mg/L for carbamazepine metabolites: 10,11-trans-dihydroxy-10,11-dihydrocarbamazepine and carbamazepine-10,11-epoxide; 0.1-10 mg/L for oxcarbazepine; 2-100 mg/L for felbamate and 3-150 mg/L for ethosuximide. The suitability of the validated method for routine therapeutic drug monitoring was confirmed by quantification of the analytes in plasma samples from patients with epilepsy on combination antiepileptic therapy.


Mineralocorticoid action and sodium-hydrogen exchange: studies in experimental cardiac fibrosis.
There is increasing evidence that the trigger for cardiac fibrosis in response to mineralocorticoid/salt administration is coronary vasculitis and that effects can be seen within days of deoxycorticosterone acetate (DOCA) administration. Furthermore, rapid, nongenomic mineralocorticoid effects on the sodium-hydrogen exchanger (NHE-1) in vascular smooth muscle cells have recently been described. That this mechanism may act as an inflammatory or profibrotic signal was tested by comparing the specific NHE-1 antagonist cariporide and the mineralocorticoid receptor antagonist K canrenoate in the rat model of mineralocorticoid/salt perivascular fibrosis over 8 d of DOCA/salt administration. Interstitial collagen, inflammatory cell infiltration, and inflammatory markers were determined. DOCA elevated blood pressure above control, cariporide +DOCA, or K canrenoate +DOCA rats, without cardiac hypertrophy. At 8 d interstitial collagen was significantly elevated in the DOCA-alone group, with levels in cariporide- and K canrenoate-treated rats not different from control. Expression of osteopontin, cyclooxygenase-2, and ED-1 were elevated by DOCA treatment, blocked by potassium canrenoate, and (for ED-1 and osteopontin) partially reduced by cariporide. These results suggest mineralocorticoid/salt-induced cardiac fibrosis may involve coronary vascular smooth muscle cell NHE-1 activity as a possible contributor to the cascade of transcriptional events that produce the characteristic coronary vasculitis seen with excess mineralocorticoid and salt.


Biochemical aspects of Salter's osteotomy for treatment of acetabular dysplasia.
The center-edge (CE) angle was measured on serial antero-posterior radiographs of 44 hips treated by Salter's innominate osteotomy for developmental dysplasia of the hip. Radiographic examination according to Severin was performed and the peak hip joint pressure was calculated using geometrical parameters determined from radiographs at the most recent follow-up examination, 7 to 13 years postoperatively. At the latest follow-up assessment, there was a positive correlation between the postoperative CE angle and radiographic results. Negative correlation was found between the postoperative CE angle and peak hip joint pressure at the latest follow up. In conclusion, Salter's osteotomy performed to treat developmental dysplasia of the hip should involve radical acetabular correction resulting in maximum increase of the CE angle.


Ethnic variation in the clinical manifestations of rheumatoid arthritis: role of HLA-DRB1 alleles.
To assess the extent of ethnic variation in the clinical expression of rheumatoid arthritis (RA) and the role of HLA-DRB1 alleles in this variation. We assessed consecutive RA patients for joint findings, subcutaneous nodules, laboratory and radiographic findings, and treatment. We typed HLA-DRB1 alleles to identify those that contain the shared epitope (SE). We adjusted ethnic comparisons for age and sex, and tested for ethnic heterogeneity in the effect of the SE. We studied 777 RA patients, 498 of whom were women (64%), 432 were Hispanic (56%), 272 were non-Hispanic white (NHW; 35%), 53 were African American (AA; 7%), and 20 were Asian (3%). Compared with NHW, Hispanics had significantly more tender joints (17 versus 11), more swollen joints (8 versus 7), more frequent rheumatoid factor (RF) positivity (93% versus 84%), higher erythrocyte sedimentation rate (ESR; 45 versus 36 mm/hr), and a lower number of lifetime disease-modifying antirheumatic drugs (1.9 versus 2.5). AA were older at onset (46 versus 44 years), had less frequent subcutaneous nodules (18% versus 28%), and higher ESR (42 versus 36 mm/hour) than did NHW. Hispanics and AA were more likely than NHW to be null for the SE (odds ratio [OR] = 4.59 for AA; and OR = 1.61 for Hispanics), and less likely to have 2 SE-carrying alleles (OR = 0.59 for Hispanics and OR = 0.25 for AA). The number of SE copies was associated with subcutaneous nodules, ESR, RF, and radiographic changes. Ethnic heterogeneity in the effect of the SE was modest. There is ethnic variation in the clinical expression of RA and in both the frequency and types of SE-carrying HLA-DRB1 alleles. Some ethnic variation in clinical findings is associated with differences in SE frequency. However, we found that the effect of the SE on the clinical features of RA varies little between ethnic groups.


Occupational Exposure to Dust Produced when Milling Thermally Modified Wood.
During production, thermally modified wood is processed using the same machining operations as unmodified wood. Machining wood is always accompanied with the creation of dust particles. The smaller they become, the more hazardous they are. Employees are exposed to a greater health hazard when machining thermally modified wood because a considerable amount of fine dust is produced under the same processing conditions than in the case of unmodified wood. The International Agency for Research on Cancer (IARC) states that wood dust causes cancer of the nasal cavity and paranasal sinuses and of the nasopharynx. Wood dust is also associated with toxic effects, irritation of the eyes, nose and throat, dermatitis, and respiratory system effects which include decreased lung capacity, chronic obstructive pulmonary disease, asthma and allergic reactions. In our research, granular composition of particles resulting from the process of longitudinal milling of heat-treated oak and spruce wood under variable conditions (i.e., the temperature of modification of 160, 180, 200 and 220 °C and feed rate of 6, 10 and 15 m.min-1) are presented in the paper. Sieve analysis was used to determine the granular composition of particles. An increase in fine particle fraction when the temperature of modification rises was confirmed by the research. This can be due to the lower strength of thermally modified wood. Moreover, a different effect of the temperature modification on granularity due to the tree species was observed. In the case of oak wood, changes occurred at a temperature of 160 °C and in the case of spruce wood, changes occurred at the temperatures of 200 and 220 °C. At the temperatures of modification of 200 and 220 °C, the dust fraction (i.e., that occurred in the mesh sieves, particles with the size ≤ 0.08 mm) ranged from 2.99% (oak wood, feed rate of 10 m.min-1) to 8.07% (spruce wood, feed rate of 6 m.min-1). Such particles might have a harmful effect on employee health in wood-processing facilities.


[Acute primary pyelonephritis or active phase of a latent form of chronic pyelonephritis?].
Analyzing the data of a comprehensive examination of 372 patients admitted to hospital with a distinct clinical picture of acute pyelonephritis, the author shows that out of these patients' population, 278 (74.8%) had an active form of chronic disease running a latent course. At the same time 40.4% of cases were diagnosed to have chronic renal failure (CRF), stages IB-IIB. To determine the genuine character of renal injury, the author thinks it necessary to regard the symptom-complex of acute pyelonephritis as a consequence of the pyelonephritic process itself. In complicated cases of the diagnosis, biopsy of the kidney concerned is required. In 176 patients the final diagnosis was established by uroradiological examination, in 59 by means of the use of laboratory methods, and in 137 by biopsy of the involved kidney. It is recommended that patients with renal pathology should be kept under prolonged dispensary observation, particularly those with an initial stage of CRF.


[Smoking in schools. A cross-sectional epidemiologic study of a population of 4,281 school children].
Smokers begin to acquire their habit early. The objective of this study was therefore to determine real smoking habits among young people in Spain and create a profile of the adolescent smoker. A questionnaire was administered to a cross-section of the population. The results obtained coincide with reports by other authors in the Spanish and international literature. We found that the prevalence of smoking among young people between 9 and 19 years of age is approximately 25% and tobacco use increases with age. Young smokers participate less in sports and have a greater tendency to approve tobacco advertising. Young smokers' reasons for starting to smoke are diverse and the notion of "being grown-up" did not appear to apply to our cohort. We discuss, in the light of recent findings, the role of parents and social context in encouraging young people to smoke. The age of 14 is a key moment for taking up and establishing the habit of smoking.


[Study of pesticide poisoning during a nursing course: report of the experience].
The aim of this paper is to describe the experience of nursing students experience in the development of a research project. It began through the identification of a public health problem in a farm, when 1,200 people camped near a storehouse containing organocloride pesticides. The group decided for a descriptive study. The students describe their participation in the project, with specialists consultation, bibliographic review, development of a research tool, capacitate people to collect data and analysis with statistical help. As a conclusion, they point out the limitations encountered, as well as the benefits obtained in this experience during undergraduate course.


Differentiation of mesenchymal stem cells under hypoxia and normoxia: lipid profiles revealed by time-of-flight secondary ion mass spectrometry and multivariate analysis.
Mesenchymal stem cells (MSC) have the ability to self-renew and differentiate into multiple cell types valuable for clinical treatment of rheumatic pathologies. To study the chondrogenic potential of MSC and identify the conditions that recreate the native cartilage environment, we used time-of-flight secondary ion mass spectrometry (TOF-SIMS) for label-free detection of cell-type- and environmental-condition-specific molecular profiles. We observed that coculture of human MSC and chondrocytes under standard culture conditions leads to improved extracellular matrix (ECM) deposition. In marked contrast, this effect was lost under low oxygen tension. This improved extracellular matrix deposition was associated with a significant decrease in lipids and in particular cholesterol under low oxygen tension as revealed by TOF-SIMS coupled to principal component analysis and discriminant analysis. We furthermore demonstrate that the higher cholesterol levels under normoxia might regulate fibroblast growth factor 1 (FGF-1) gene expression which was previously implemented in increased ECM production in the cocultures. In conclusion, our study shows an unexpected role of lipids as orchestrators of chondrogenesis in response to oxygen tension which is, at least in part, mediated through FGF-1.


Effect of wheelchair design on wheeled mobility and propulsion efficiency in less-resourced settings.
Wheelchair research includes both qualitative and quantitative approaches, primarily focuses on functionality and skill performance and is often limited to short testing periods. This is the first study to use the combination of a performance test (i.e. wheelchair propulsion test) and a multiple-day mobility assessment to evaluate wheelchair designs in rural areas of a developing country. Test the feasibility of using wheel-mounted accelerometers to document bouts of wheeled mobility data in rural settings and use these data to compare how patients respond to different wheelchair designs. A quasi-experimental, pre- and post-test design was used to test the differences between locally manufactured wheelchairs (push rim and tricycle) and an imported intervention product (dual-lever propulsion wheelchair). A one-way repeated measures analysis of variance was used to interpret propulsion and wheeled mobility data. There were no statistical differences in bouts of mobility between the locally manufactured and intervention product, which was explained by high amounts of variability within the data. With regard to the propulsion test, push rim users were significantly more efficient when using the intervention product compared with tricycle users. Use of wheel-mounted accelerometers as a means to test user mobility proved to be a feasible methodology in rural settings. Variability in wheeled mobility data could be decreased with longer acclimatisation periods. The data suggest that push rim users experience an easier transition to a dual-lever propulsion system.


[Thrombolysis with rt-PA in children with arterial and venous thromboses--a new therapy concept].
Thrombolytic therapy usually used for thrombosis in the adult has been administered as a therapeutic regiment in pediatric patients (parental consent was sought prior to the treatment with rt-PA). We report our experience with rt-PA in 17 children and adolescents suffering from arterial (n = 4) or venous thrombosis (n = 13) due to local rhabdomyosarcoma, acute lymphoblastic leukemia, chronic myeloblastosis, sickle cell anaemia, parenteral nutrition, haemolytic uremic syndrome, central arterial and venous catheters and septicemia Thrombotic diseases have been diagnosed by Doppler ultrasound, computed tomography, angiography and phlebography. Rt-PA therapy was started immediately after diagnostic procedures had been performed. Rt-PA dose varied from 0.2 mg as a single dose to 0.8 mg/kg bw/d over a three day period in children local thrombolysis was performed. In patients requiring systemic thrombolytic therapy rt-PA was administered from 0.8 mg/kg bw/d in three days to 2.0 mg/kg bw/d over a whole period of three weeks in both groups during thrombolysis low dose heparin was added. When rt-PA infusion was terminated heparin (70 IU - 400 IU/kg bw/d) was administered for 7 to 14 days in order to prevent reocclusion. Later prophylaxis with coumarin derivatives in venous thrombosis and antiplatelet agents in arterial occlusive diseases was performed. In no patient did we see a decrease of fibrinogen and plasminogen during rt-PA therapy.(ABSTRACT TRUNCATED AT 250 WORDS)


Surface Water Microbial Community Response to the Biocide 2,2-Dibromo-3-Nitrilopropionamide, Used in Unconventional Oil and Gas Extraction.
Production of unconventional oil and gas continues to rise, but the effects of high-density hydraulic fracturing (HF) activity near aquatic ecosystems are not fully understood. A commonly used biocide in HF, 2,2-dibromo-3-nitrilopropionamide (DBNPA), was studied in microcosms of HF-impacted (HF+) versus HF-unimpacted (HF-) surface water streams to (i) compare the microbial community response, (ii) investigate DBNPA degradation products based on past HF exposure, and (iii) compare the microbial community response differences and similarities between the HF biocides DBNPA and glutaraldehyde. The microbial community responded to DBNPA differently in HF-impacted versus HF-unimpacted microcosms in terms of the number of 16S rRNA gene copies quantified, alpha and beta diversity, and differential abundance analyses of microbial community composition through time. The differences in microbial community changes affected degradation dynamics. HF-impacted microbial communities were more sensitive to DBNPA, causing the biocide and by-products of the degradation to persist for longer than in HF-unimpacted microcosms. A total of 17 DBNPA by-products were detected, many of them not widely known as DBNPA by-products. Many of the brominated by-products detected that are believed to be uncharacterized may pose environmental and health impacts. Similar taxa were able to tolerate glutaraldehyde and DBNPA; however, DBNPA was not as effective for microbial control, as indicated by a smaller overall decrease of 16S rRNA gene copies/ml after exposure to the biocide, and a more diverse set of taxa was able to tolerate it. These findings suggest that past HF activity in streams can affect the microbial community response to environmental perturbation such as that caused by the biocide DBNPA.IMPORTANCE Unconventional oil and gas activity can affect pH, total organic carbon, and microbial communities in surface water, altering their ability to respond to new environmental and/or anthropogenic perturbations. These findings demonstrate that 2,2-dibromo-3-nitrilopropionamide (DBNPA), a common hydraulic fracturing (HF) biocide, affects microbial communities differently as a consequence of past HF exposure, persisting longer in HF-impacted (HF+) waters. These findings also demonstrate that DBNPA has low efficacy in environmental microbial communities regardless of HF impact. These findings are of interest, as understanding microbial responses is key for formulating remediation strategies in unconventional oil and gas (UOG)-impacted environments. Moreover, some DBNPA degradation by-products are even more toxic and recalcitrant than DBNPA itself, and this work identifies novel brominated degradation by-products formed.


New developments in the use of peptide gonadotropin-releasing hormone antagonists versus agonists.
Gonadotropin-releasing hormone (GnRH) stimulates the pituitary secretion of both luteinising hormone (LH) and follicle-stimulating hormone (FSH), and thus controls the hormonal and reproductive functions of the gonads. The blockade of the effects of GnRH may be sought for a variety of reasons; for example, to control premature LH surges and to reduce the cancellation rate with the aim of improving the pregnancy rate per treatment cycle or in the treatment of sex hormone-dependent disorders. Selective blockade of LH/FSH secretion and subsequent chemical castration have previously been achieved by desensitising the pituitary to continuously administered GnRH or by giving long-acting GnRH agonists. GnRH analogues are indicated for clinical situations in which the suppression of endogenous gonadotropins (precocious puberty, contraception and controlled ovarian hyperstimulation) or sexual steroids (endometriosis, prostate hyperplasia, cancer and uterine fibroids) is desired. The immediate suppression of the pituitary that is achieved by GnRH antagonists without an initial stimulatory effect is the main advantage of these compounds over the agonists. GnRH antagonists have been developed for clinical use with acceptable pharmacokinetic, safety and commercial profiles. In assisted reproduction, these compounds seem to be as effective as established therapy, but with shorter treatment times, less use of gonadotropic hormones, improved patient acceptance, and fewer follicles and oocytes. All of the current indications for GnRH agonist desensitisation may prove to be indications for a GnRH antagonist, including endometriosis, leiomyoma and breast cancer in women, benign prostatic hypertrophy and prostatic carcinoma in men, and central precocious puberty in children. However, the best clinical evidence has been in assisted reproduction and prostate cancer.


Novel layer-by-layer interfacial [Ni(salen)]-polyelectrolyte hybrid films.
A novel multilayer film containing a cationic phosphonium-derivatized Ni(salen)-type complex and poly(sodium-4-styrenesulfonate (NaPSS) was assembled onto quartz, mica, and metal surfaces using the layer-by-layer (LbL) technique. Spectroscopic (UV-vis) and gravimetric (QCM) responses for the multilayer films show regular stepwise growth and the signature of strong electrostatic interactions between the component layers. The gravimetric responses indicate the presence of substantial additional (net neutral) material in the PSS layers, which XPS shows is not polyelectrolyte or salt, so charge compensation is intrinsic; we deduce the presence of space-filling solvent. Direct electrostatic interaction of the two-component layers is enhanced by a secondary noncovalent interaction between the delocalized pi-systems of the two components. Permeability of the film to the redox probe [Fe(CN)(6)](3-/4-) was studied by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Qualitatively similar results were obtained in the absence and presence of a precursor PSS/PAH multilayer, but with a general shift in kinetic and diffusional processes to longer time scales (lower frequencies) in the presence of the precursor layer and with increasing numbers of PSS/[Ni(salen)] bilayers. Quantitatively, the EIS data were interpreted using a capillary membrane model (CMM) to yield values of coverage, apparent charge transfer resistance, double-layer capacitance, pore size, and diffusion coefficient. The coverage values were consistent with a model in which there are no preferential growth sites and the surface charge density is independent of the number of bilayers.


Psychometric functions and temporal integration in electric hearing.
Temporal-integration functions and psychometric functions for detection were obtained in eight users of the Nucleus 22-electrode cochlear implant. Stimuli were 100-Hz, 200-microseconds/phase trains of biphasic pulses with durations ranging from 0.44 to 630.4 ms (1 to 64 pulses). Temporal-integration functions were measured for 21 electrodes. Slopes of these functions were considerably shallower than the 2.5 dB/doubling slopes typically observed in acoustic hearing. They varied widely across subjects and for different electrodes in a given subject, ranging from 0.06 to 1.94 dB/doubling of stimulus pulses, with a mean [standard deviation (s.d.)] value of 0.42 (0.38). Psychometric functions were measured for 11 of the same 21 electrodes. Slopes of psychometric functions also varied across subjects and electrodes, and were 2-20 times steeper than those reported by other investigators for normal-hearing and cochlear-impaired acoustic listeners. Slopes of individual psychometric functions for 1-, 2-, 4-, and 8-pulse stimuli ranged from 0.20 to 1.84 log d'/dB with a mean (s.d.) value of 0.77 (0.45). Psychometric-function slopes did not vary systematically with stimulus duration in most cases. A clear inverse relation between slopes of psychometric functions and slopes of temporal-integration functions was observed. This relation was reasonably well described by a hyperbolic function predicted by the multiple-looks model of temporal integration [Viemeister and Wakefield, J. Acoust. Soc. Am. 90, 858-865 (1991)]. Psychometric-function slopes tended to increase with absolute threshold and were inversely correlated with dynamic range, suggesting that observed differences in psychometric-function slopes across subjects and electrodes may reflect underlying differences in neural survival.


CD45 ligation induces programmed cell death in T and B lymphocytes.
Studies have shown an essential role for the CD45 protein tyrosine phosphatase in regulating Ag receptor-derived signals in lymphocytes. Co-ligating CD45 with the Ag receptor, however, can also inhibit receptor signaling, as manifest in T cells by the lack of IL-2 production and proliferation. We report that CD45 ligation alone induces apoptosis in normal T cells, and that death was greatly potentiated by cross-linking CD3. Normal B cells and T and B cell lines were also induced to die with insoluble CD45 mAb. CD45-induced cell death was blocked by inhibitors of protein tyrosine kinases and protein tyrosine phosphatases, but not by inhibitors of RNA or protein synthesis or by cyclosporin A. Morphologically, CD45-mediated apoptosis resembled death induced via CD95 (Fas), as evidenced by nuclear condensation and membrane blebbing, but did not cause DNA fragmentation into oligonucleosomes. Co-ligating CD45 and CD95 either enhanced or inhibited CD45-induced cell death, depending on the degree of CD45 and CD95 cross-linking. Finally, CD45 cross-linking induced its rapid association with the detergent-insoluble cell fraction, suggesting that it becomes linked to the cytoskeleton during CD45-induced apoptosis. These data show a novel role for CD45 in regulating lymphocyte death as well as proliferation.