The roles of type 2 diabetes and obesity in disease activity and progression of non-alcoholic fatty liver disease/non-alcoholic steatohepatitis.
We examined the roles of type 2 diabetes (T2D) and obesity in disease activity and fibrosis progression/regression in patients with non-alcoholic fatty liver disease/non-alcoholic steatohepatitis (NAFLD/NASH). This multi-center, retrospective study included patients with suspected or histologically proven NAFLD/NASH from the NASH Clinical Research Network. Outcomes included disease activity and rate of fibrosis, assessed using liver-biopsy driven measures (NAFLD activity score [NAS] and fibrosis score [FS]). Logistic regression and doubly robu estimation of causal effects tested relationships among T2D, obesity, and NAFLD/NASH. The analytical sample included 870 adult patients with baseline biopsy data and 157 patients with multiple biopsy data. Patients with NAFLD/NASH and T2D had significantly higher baseline average NAS (4.52 vs. 4.13; p = 0.009) and FS (2.17 vs. 1.56; p < 0.0001); those with T2D had a significantly greater reduction in average NAS over time (-0.77/year vs. -0.17/year; p = 0.0008). Change in FS over time did not differ significantly by T2D status (-0.23/year vs. -0.04/year; p = 0.34). Baseline NAS, baseline FS, and change in average NAS over time did not differ significantly by obesity status (4.17 vs. 4.47; p = 0.16; 1.73 vs.1.92; p = 0.31; -0.40/year vs. -0.59/year; p = 0.62, respectively). Patients with obesity had a slight increase in FS but those without obesity had a reduction in average FS over time (0.07/year vs. -0.27/year; p = 0.008). Patients with NAFLD/NASH and T2D had greater baseline disease activity versus those without T2D, but there was greater regression of disease activity over time among those with T2D. Patients with NAFLD/NASH and obesity had worsening of fibrosis versus those without obesity. NCT00063622.


Should You Recommend Inhaled Corticosteroids for This Patient With Chronic Obstructive Pulmonary Disease?: Grand Rounds Discussion From Beth Israel Deaconess Medical Center.
Approximately 12 million adults in the United States receive a diagnosis of chronic obstructive pulmonary disease (COPD) each year, and it is the fourth leading cause of death. Chronic obstructive pulmonary disease refers to a group of diseases that cause airflow obstruction and a constellation of symptoms, including cough, sputum production, and shortness of breath. The main risk factor for COPD is tobacco smoke, but other environmental exposures also may contribute. The GOLD (Global Initiative for Chronic Obstructive Lung Disease) 2020 Report aims to provide a nonbiased review of the current evidence for the assessment, diagnosis, and treatment of patients with COPD. To date, no conclusive evidence exists that any existing medications for COPD modify mortality. The mainstay of treatment for COPD is inhaled bronchodilators, whereas the role of inhaled corticosteroids is less clear. Inhaled corticosteroids have substantial risks, including an increased risk for pneumonia. Here, 2 experts, both pulmonologists, reflect on the care of a woman with severe COPD, a 50-pack-year smoking history, frequent COPD exacerbations, and recurrent pneumonia. They consider the indications for inhaled corticosteroids in COPD, when inhaled corticosteroids should be withdrawn, and what other treatments are available.


Widespread Genetic Incompatibilities between First-Step Mutations during Parallel Adaptation of Saccharomyces cerevisiae to a Common Environment.
Independently evolving populations may adapt to similar selection pressures via different genetic changes. The interactions between such changes, such as in a hybrid individual, can inform us about what course adaptation may follow and allow us to determine whether gene flow would be facilitated or hampered following secondary contact. We used Saccharomyces cerevisiae to measure the genetic interactions between first-step mutations that independently evolved in the same biosynthetic pathway following exposure to the fungicide nystatin. We found that genetic interactions are prevalent and predominantly negative, with the majority of mutations causing lower growth when combined in a double mutant than when alone as a single mutant (sign epistasis). The prevalence of sign epistasis is surprising given the small number of mutations tested and runs counter to expectations for mutations arising in a single biosynthetic pathway in the face of a simple selective pressure. Furthermore, in one third of pairwise interactions, the double mutant grew less well than either single mutant (reciprocal sign epistasis). The observation of reciprocal sign epistasis among these first adaptive mutations arising in the same genetic background indicates that partial postzygotic reproductive isolation could evolve rapidly between populations under similar selective pressures, even with only a single genetic change in each. The nature of the epistatic relationships was sensitive, however, to the level of drug stress in the assay conditions, as many double mutants became fitter than the single mutants at higher concentrations of nystatin. We discuss the implications of these results both for our understanding of epistatic interactions among beneficial mutations in the same biochemical pathway and for speciation.


Statistical Analysis of Influences on the Psychosocial Status of Children With Hemangiomas and Their Families.
The psychologic stress on the child and family, which arise from hemangiomas, the most common neoplasm of childhood, cannot be overestimated. This study determined the preoperative and postoperative psychosocial status and variation among Oriental children with hemangiomas and their families by questionnaire. Thirty patients who underwent surgery for hemangiomas were assessed for preoperative and postoperative psychosocial status by questionnaire. The distribution of the total mean score and variation between the preoperative and postoperative status was estimated. Based on these results, the significance was statistically analyzed according to variable determinants. This study showed that hemangiomas have harmful effects on psychosocial status of patients and families. After corrective surgery, an improvement in psychosocial status was noted with respect to the self-esteem category or categories related to social activity, and in the following variables, women, face, and dissatisfaction with appearance. When the authors care for patients with hemangiomas and their families, the psychosocial health must be presumed to be at particular risk. Earlier surgical interventions with esthetic concerns have permitted the patient and family the opportunity to reduce the psychologic impact that the hemangioma may otherwise have.


Quantifying test-retest reliability of repeated objective attentional measures in Lewy body dementia.
Objective cognitive impairment is a feature of Lewy body dementia (LBD), and computerised attentional tasks are commonly used as outcome measures in interventional trials. However, the reliability of these measures, in the absence of interventions, are unknown. This study examined the reliability of these attentional measures at short-term and longer-term follow-up stages. LBD patients (n = 36) completed computerised attentional tasks [simple and choice reaction time, and digit vigilance (SRT, CRT, DV)] at short-term (Day 0-Day 5) and longer-term (4 and 12 weeks) follow-up. Intra-class correlations (ICCs) were calculated to assess test-retest reliability. At short-term, the reciprocal SRT, CRT and DV mean reaction time to correct answers, the reciprocal DV coefficient of variation, and reciprocal power of attention (PoA) all showed excellent levels of reliability (all ICCs > 0.90). The reciprocal PoA showed the highest level of reliability (ICC = 0.978). At longer-term follow-up, only the reciprocal PoA had excellent levels of reliability (ICC = 0.927). Reciprocal SRT, CRT and DV reaction time to correct answers, and the CRT coefficient of variation values, showed good levels of test-retest reliability (ICCs ≥ 0.85). Contrary to expectations, most attentional measures demonstrated high levels of test-retest reliability at both short-term and longer-term follow-up time points. The reciprocal PoA composite measure demonstrated excellent levels of test-retest reliability, both in the short-term and long-term. This indicates that objective attentional tasks are suitable outcome measures in LBD studies and that the composite PoA measure may offer the highest levels of reliability.


Improved watershed transform for medical image segmentation using prior information.
The watershed transform has interesting properties that make it useful for many different image segmentation applications: it is simple and intuitive, can be parallelized, and always produces a complete division of the image. However, when applied to medical image analysis, it has important drawbacks (oversegmentation, sensitivity to noise, poor detection of thin or low signal to noise ratio structures). We present an improvement to the watershed transform that enables the introduction of prior information in its calculation. We propose to introduce this information via the use of a previous probability calculation. Furthermore, we introduce a method to combine the watershed transform and atlas registration, through the use of markers. We have applied our new algorithm to two challenging applications: knee cartilage and gray matter/white matter segmentation in MR images. Numerical validation of the results is provided, demonstrating the strength of the algorithm for medical image segmentation.


Amyloid neuropathy and multiple myeloma. Ultrastructural and immunopathological study of two cases.
Report of 2 patients suffering from sensory peripheral neuropathy and multiple myeloma. In 1 case, peripheral neuropathy occurred several months before the appearance of multiple myeloma. A peripheral nerve biopsy exhibited numerous amyloid deposits within the endoneurium. Amyloid deposits have been estimated to occur in 15% of cases of multiple myeloma, but only 4 other detailed observations of amyloid deposits within the endoneurium have been reported in the literature. Other mechanisms might be involved since most peripheral neuropathies associated with multiple myeloma do not exhibit any amyloid deposits.


Identification of pararosaniline as a modifier of RNA splicing in Caenorhabditis elegans.
Posttranscriptional splicing of premessenger RNA (mRNA) is an evolutionarily conserved eukaryotic process for producing mature mRNA that is translated into proteins. Accurate splicing is necessary for normal growth and development, and aberrant splicing is increasingly evident in various human pathologies. To study environmental factors that influence RNA splicing, we employed a fluorescent Caenorhabditis elegans in vivo splicing reporter as a biomarker for splicing fidelity to screen against the US EPA ToxCast chemical library. We identified pararosaniline hydrochloride as a strong modifier of RNA splicing. Through gene expression analysis, we found that pararosaniline activates the oxidative stress response and alters the expression of key RNA splicing regulator genes. Physiological assays show that pararosaniline is deleterious to C. elegans development, reproduction, and aging. Through a targeted RNAi screen, we found that inhibiting protein translation can reverse pararosaniline's effect on the splicing reporter and provide significant protection against long-term pararosaniline toxicity. Together, this study reveals a new chemical modifier of RNA splicing and describes translation inhibition as a genetic mechanism to provide resistance.


Oscillatory activity, phase differences, and phase resetting in the inferior olivary nucleus.
The generation of temporal patterns is one of the most fascinating functions of the brain. Unlike the response to external stimuli temporal patterns are generated within the system and recalled for a specific use. To generate temporal patterns one needs a timing machine, a "master clock" that determines the temporal framework within which temporal patterns can be generated and implemented. Here we present the concept that in this putative "master clock" phase and frequency interact to generate temporal patterns. We define the requirements for a neuronal "master clock" to be both reliable and versatile. We introduce this concept within the inferior olive nucleus which at least by some scientists is regarded as the source of timing for cerebellar function. We review the basic properties of the subthreshold oscillation recorded from olivary neurons, analyze the phase relationships between neurons and demonstrate that the phase and onset of oscillation is tightly controlled by synaptic input. These properties endowed the olivary nucleus with the ability to act as a "master clock."


Targeting the perpetrator: breast cancer stem cell therapeutics.
The hypothesis that tumors may originate from a rare population of cancer stem cells (CSCs) has gained tremendous popularity in recent years and is supported extensively by several pioneering works. Cancer therapies targeting CSCs have unlimited potential for relapse free survival of cancer patients. As a result, knowledge of biological pathways that govern CSCs is very important and this review is focused on the biology of CSCs with special emphasis on breast CSCs, and recent advances in therapeutic approaches targeting them.


Comparison of internal anterior chamber diameter imaging modalities: 35-MHz ultrasound biomicroscopy, Visante optical coherence tomography, and Pentacam.
To compare the internal diameter of the anterior chamber among three different imaging modalities: 35-MHz ultrasound biomicroscopy (UBM), Visante optical coherence tomography (OCT), and Pentacam, and to compare the internal diameters with white-to-white (WTW) distance measured by Orbscan. The internal horizontal anterior chamber diameter was measured in 20 normal eyes using 35-MHz UBM (Optikon), Visante OCT (Carl Zeiss Meditec), and Pentacam (Oculus Inc). The mean horizontal anterior chamber diameters were compared, and the correlations between each pair of two different methods were evaluated. Horizontal WTW distance was measured by Orbscan IIz (Bausch & Lomb-Orbtek Inc) and then correlated with the measurements of internal anterior chamber diameter. The mean horizontal anterior chamber diameter was 10.99+/-0.47 mm with UBM, 11.70+/-0.47 mm with OCT, and 11.86+/-0.45 mm with Pentacam. Ultrasound biomicroscopy measurements were significantly different than those of OCT and Pentacam (P<.001 for both). However, no significant difference was found between OCT and Pentacam (P=.19). Regarding the correlations between each pair of two different methods, UBM and OCT showed significant correlation (r=0.848, P<.001), but there was no significant correlation between OCT and Pentacam (r=0.403, P=.078), even though the mean values were similar. In the Bland-Altman plots, the range of agreement was 0.51 mm between OCT and UBM, and 0.99 mm between Pentacam and OCT. Although the internal anterior chamber diameters measured by 35-MHz UBM were significantly smaller than those measured by Visante OCT and Pentacam, the measurements of UBM and Visante OCT showed significant correlation and good agreement with each other. Pentacam showed poor correlation with other measurements.


Tuning Liposome Membrane Permeability by Competitive Peptide Dimerization and Partitioning-Folding Interactions Regulated by Proteolytic Activity.
Membrane active peptides are of large interest for development of drug delivery vehicles and therapeutics for treatment of multiple drug resistant infections. Lack of specificity can be detrimental and finding routes to tune specificity and activity of membrane active peptides is vital for improving their therapeutic efficacy and minimize harmful side effects. We describe a de novo designed membrane active peptide that partition into lipid membranes only when specifically and covalently anchored to the membrane, resulting in pore-formation. Dimerization with a complementary peptide efficiently inhibits formation of pores. The effect can be regulated by proteolytic digestion of the inhibitory peptide by the matrix metalloproteinase MMP-7, an enzyme upregulated in many malignant tumors. This system thus provides a precise and specific route for tuning the permeability of lipid membranes and a novel strategy for development of recognition based membrane active peptides and indirect enzymatically controlled release of liposomal cargo.


The induction and inhibition of differentiation in normal and leukaemic cells.
For granulocytic-macrophage progenitor populations and their progeny, five glycoproteins have been identified: GM-CSF, G-CSF, multi-CSF, M-CSF and IL-6 that can regulate their proliferative activity, maturation and functional activities. The same glycoproteins also have a capacity to induce irreversible differentiation commitment in normal bipotential granulocyte-macrophage progenitors and in some myeloid leukaemic cell lines, which suggests that common cellular processes exist in both situations. The leukaemia inhibitory factor (LIF) is a glycoprotein, with intriguing properties, which can either induce differentiation in some myeloid leukaemic cell lines or prevent differentiation in normal totipotential embryonic stem cells. The data from the LIF studies suggest a genetic mechanism controlling self-generation that is relatively simple and may be common to all cells. However, the actual cellular response observed appears to depend on the nature of the responding cell.


Efficacy of an Acute Pain Titration Protocol Driven by Patient Response to a Simple Query: Do You Want More Pain Medication?
We assess the efficacy of a simple pain titration protocol of 1-mg increments of intravenous hydromorphone, given at fixed intervals, driven solely by patient response to a yes/no question. This was a prospective interventional cohort study of nonelderly adults with acute severe pain defined as requiring intravenous opioids in the judgment of the attending emergency physician. All patients received 1 mg intravenous hydromorphone and 30 minutes later were asked, "Do you want more pain medication?" Patients responding yes received an additional 1 mg of intravenous hydromorphone and were asked the same question 30 minutes after receiving it. Those responding no did not receive additional opioid and were asked the question again 30 minutes later. Each patient was queried 4 times. The primary endpoint was the proportion of patients achieving satisfactory pain control, defined as declining additional pain medication on 1 or more occasions. Of 215 patients enrolled, there were 8 protocol violations, leaving 207 patients with analyzable data; 205 of 207 patients (99%; 95% confidence interval 97% to 100%) achieved satisfactory analgesia at 1 or more points during the study. Nine patients desaturated below 95% on room air, 2 had respiratory rates less than 10 breaths/min, and 2 had pulse rates less than 50 beats/min. No adverse events were associated with amount of hydromorphone received. A pain protocol, based on titration of 1 mg intravenous hydromorphone, driven solely by patient response to a simple standardized question repeated at intervals, resulted in achievement of satisfactory analgesia on at least 1 occasion in 99% of patients.


Pay for Performance in Medicaid: Evidence from Three Natural Experiments.
To examine the impact of pay for performance in Medicaid on the quality and utilization of care. Medicaid claims and encounter data in three intervention states (Pennsylvania, Minnesota, and Alabama) and three comparison states. Difference-in-difference analysis with propensity score-matched comparison group. Primary outcomes of interest were Healthcare Effectiveness Data and Information Set (HEDIS)-like process measures of quality, utilization by service category, and ambulatory care-sensitive admissions and emergency department visits. In Pennsylvania, there was a statistically significant reduction of 88 ambulatory visits per 1,000 enrollee months compared with Florida. In Minnesota, there was a significant decrease of 7.2 hospital admissions per thousand enrollee months compared with Wisconsin. In Alabama, where incentives were not paid out until the end of a 2-year waiver period, there was a decline of 1.6 hospital admissions per thousand member months, and an increase of 59 ambulatory visits per 1,000 enrollees compared with Georgia. No significant quality improvements in intervention relative to control states. Our findings are mixed, with no measurable quality improvements across the three states, but reductions in hospital admissions in two programs. As states move to value-based payment for patient-centered medical homes and Accountable Care Organizations, lessons learned from these pioneering states should inform program design.


The preparation of monoclonal antibodies to human bone and liver alkaline phosphatase and their use in immunoaffinity purification and in studying these enzymes when present in serum.
1. Liver and bone alkaline phosphatase isoenzymes were solubilized with the zwitterionic detergent sulphobetaine 14, and purified to homogeneity by using a monoclonal antibody previously raised against a partially-purified preparation of the liver isoenzyme. Both purified isoenzymes had a specific activity in the range 1100-1400 mumol/min per mg of protein with a subunit Mr of 80,000 determined by SDS/polyacrylamide gel electrophoresis. Butanol extraction instead of detergent solubilization, before immunoaffinity purification of the liver enzyme, resulted in the same specific activity and subunit Mr. The native Mr of the sulphobetaine 14-solubilized enzyme was consistent with the enzyme being a dimer of two identical subunits and was higher than that of the butanol-extracted enzyme, presumably due to the binding of the detergent micelle. 2. Pure bone and liver alkaline phosphatase were used to raise further antibodies to the two isoenzymes. Altogether, 27 antibody-producing cell lines were cloned from 12 mice. Several of these antibodies showed a greater than 2-fold preference for bone alkaline phosphatase in the binding assay used for screening. No antibodies showing a preference for liver alkaline phosphatase were successfully cloned. None of the antibodies showed significant cross-reaction with placental or intestinal alkaline phosphatase. Epitope analysis of the 27 antibodies using liver alkaline phosphatase as antigen gave rise to six groupings, with four antibodies unclassified. The six major epitope groups were also observed using bone alkaline phosphatase as antigen. 3. Serum from patients with cholestasis contains soluble and particulate forms of alkaline phosphatase. The soluble serum enzyme had the same size and charge as butanol-extracted liver enzyme on native polyacrylamide-gel electrophoresis. Cellulose acetate electrophoresis separated the soluble and particulate serum alkaline phosphatases as slow- and fast-moving forms respectively. In the presence of sulphobetaine 14 all the serum enzyme migrated as the slow-moving form on cellulose acetate electrophoresis. Monoclonal anti-(alkaline phosphatase) immunoadsorbents did not bind the particulate form of alkaline phosphatase in cholestatic serum but bound the soluble form. In the presence of sulphobetaine 14 all the cholestatic serum alkaline phosphatase bound to the immunoadsorbents. 4. The electrophoretic and immunological data are consistent with both particulate and soluble forms of alkaline phosphatase in cholestatic serum being derived from the hepatocyte membrane.


Assessment of exposure to pesticides: residues in 24 h duplicate diets versus their metabolites in 24 h urine using suspect screening and target analysis.
Human biomonitoring can add value to chemical risk assessment by reducing the assumptions regarding consumption rates, residue occurrence, and processing effects and by integrating exposures from different sources (diet, household use, environmental). However, the relationship between exposure and concentration in human matrices is unknown for most pesticides. Therefore, we conducted a pilot study to gain more insight into the qualitative and quantitative relationship between dietary intake of pesticides (external exposure) and urinary excretion (reflecting internal exposure). In this cross-sectional observational study, 35 healthy consumers aged 18-65 years from the region of Wageningen, Netherlands, collected an exact duplicate portion of their diets during 24 h. On the same day, they also collected all their urine. The duplicate diets were analyzed using target screening by GC- and LC-HRMS; each duplicate diet contained at least five, up to 21, pesticide residues. The 24 h urine samples were analyzed using LC-HRMS in a suspect screening workflow. Metabolites were tentatively detected in all 24 h urine samples, ranging from six metabolites corresponding to four pesticides up to 40 metabolites originating from 16 pesticides in a single urine sample. In total, 65 metabolites originating from 28 pesticides were tentatively detected. After prioritization and additional confirmation experiments, 28 metabolites originating from 10 pesticides were identified with confidence level 1 or 2b. Next, quantitative analysis was performed for a selection of pesticides in duplicate diets and their metabolites in 24 h urine to assess quantitative relationships. In the quantitative comparisons between duplicate diet and 24 h urine, it was found that some metabolites were already present in the duplicate diet, which may give an overestimation of exposure to the parent pesticide based on measurement of the metabolites in urine. Additionally, the quantitative comparisons suggest a background exposure through other exposure routes. We conclude that suspect screening of 24 h urine samples can disclose exposure to mixtures of pesticide on the same day in the general population. However, more research is needed to obtain quantitative relationships between dietary intake and exposure.


MHC-restricted recognition of autologous melanoma by tumor-specific cytotoxic T cells. Evidence for restriction by a dominant HLA-A allele.
Autologous melanoma-specific CTL recognize a common tumor-associated Ag (TAA) in the context of HLA class I antigens. We have demonstrated that HLA-A2 can be a restricting Ag and, in T cell lines homozygous for HLA-A2, that CTL can be generated by stimulation with HLA-A2 allogeneic melanomas. In the current study, we have investigated T cell lines from patients who are heterozygous at HLA-A region locus, to determine the relative importance of each A-region allele in this MHC-restricted recognition of tumor. We have shown that HLA-A1 can be a restricting Ag, and that allogeneic melanomas expressing HLA-A1 can substitute for the autologous tumor in the generation of HLA-A1-restricted CTL. However, when T cell lines express both HLA-A1 and HLA-A2, the HLA-A2 allele governed restriction of the melanoma TAA. Three autologous-stimulated HLA-A1, A2 CTL lines all demonstrated restriction by the HLA-A2 allele, when examined in cytotoxicity assays, cold-competition assays, and proliferation assays. There was no evidence of restriction by the second HLA-allele, HLA-A1. Although the autologous-stimulated CTL use a single A-region allele for tumor recognition, the autologous HLA-A1, A2 tumors are lysed by both HLA-A1-restricted and HLA-A2-restricted CTL. The dominance of restricting alleles was further demonstrated when HLA-matched allogeneic melanomas were used as the stimulating tumor to generate tumor-specific CTL. Stimulation of the heterozygous (HLA-A1, A2) lymphocytes with HLA-A2-matched allogeneic melanomas resulted in CTL specific for the autologous tumor, and restricted by the HLA-A2 Ag. However, stimulation with an HLA-A1-matched allogeneic melanoma failed to induce tumor-specific CTL restricted by the HLA-A1 Ag. The data suggest there is a dominance of HLA-A region Ag at the level of the T cell, such that only one is restricting in the recognition of the autologous melanoma. At the level of the tumor, however, the TAA is expressed in the context of both HLA-A region alleles. We can generate specific CTL from lymph node cells or PBL and HLA-A region matched allogeneic melanomas; however, because most patients are heterozygous at the HLA-A region locus, an understanding of the dominant restricting alleles must be obtained so that an appropriately matched allogeneic melanoma can be selected.


Construction of a hybrid quadrupole/Fourier transform ion cyclotron resonance mass spectrometer for versatile MS/MS above 10 kDa.
Technological advancements including an open-cylindrical Penning trap with capacitively coupled ICR cell, selective ion accumulation with a resolving quadrupole, and a voltage gradient used during ion extraction from an octopole ion trap, have individually improved dynamic range and sensitivity in Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (FT-ICR MS). Documented here is a new instrument utilizing these technologies toward the robust detection and fragmentation of biomolecules >10 kDa. Up to 55-fold enhancement in ion population by selective ion accumulation combined with 10- to 20- fold signal-to-noise improvement by application of a DC voltage gradient to an accumulation octopole during the ion transfer event offers improved signal-to-noise (or speed) of MS/MS experiments, for proteins from Methanococcus jannaschii and Saccharomyces cerevisiae whole cell lysates. After external quadrupole filtering with a 40 m/z window, three proteins were fragmented (and identified) in parallel from the database of Methanococcus jannaschii. Electron capture dissociation (ECD) of an intact yeast protein provides extensive sequence information resulting in a high degree of localization for an N-terminal acetylation. Hybrid fragmentation, infrared multiphoton dissociation (IRMPD) followed by low energy electrons (ECD), with the electron source located laterally off the z-axis and external to the magnet bore, presents a strategy for identification of proteins by means of the sequence tag approach. Automated implementation of diverse MS(n) approaches in a Q-FTMS instrument promises to help realize "top-down" proteomics in the future.


Antioxidant and Antiinflammatory Activity of Vitex negundo.
Reactive oxygen species are implicated in various inflammatory disorders. Vitex negundo is mentioned in Ayurveda as useful in treating arthritic disorders. The present work was undertaken to evaluate the antioxidant potential and anti-inflammatory activity of the plant. The total methanol extract of the plant was standardized in terms of total polyphenols. The standardized extract in a dose of 100 mg/kg caused a comparable reduction in edema with that of diclofenac sodium (25 mg/kg) when evaluated for antiinflammatory activity by carrageenan-induced rat paw edema method. The extract also exhibited a strong free radical scavenging activity by 1,1-diphenyl-2-picrylhydrazyl method and caused a significant reduction in the formation of thiobarbituric acid reacting substances when evaluated for its lipid peroxidation inhibitory activity. The results strongly suggest that radical quenching may be one of the mechanisms responsible for its antiinflammatory activity.


Comparative costs of diagnosis and treatment in acute pharyngitis.
Throat cultures from 86 consecutive patients with a clinical diagnosis of pharyngitis yielded 23 (27%) that were positive for beta-hemolytic streptococci. The cost of using oral antibiotics and of obtaining and processing throat cultures was determined. Using these data the following two theoretic strategies were cost accounted: Strategy A: Empiric Treatment. Obtain no cultures but treat all patients with oral antibiotics for ten days. Total cost = the cost of antibiotics for 86 patients. Strategy B? Selective Treatment. Obtain throat cultures on all patients. Treat patients have cultures positive for beta-hemolytic streptococci with a ten-day course of oral antibiotics. Costs were similar for the two strategies. Culture costs are itemized, and methods for cost reduction are suggested. This study presents economic justification for selective treatment as the appropriate approach in the majority of cases of acute pharyngitis of undetermined cause.


AIDS and sex education for young people in China.
Although China has had a rich sexual culture for thousands of years, Chinese people are usually unwilling to openly discuss issues of sex. Some parents are quite ignorant of the change in their children's sexual attitude and behaviour. In China today, adolescents are becoming much more sexually liberated. Premarital sex and unplanned pregnancies among teenagers are increasing. Sexually transmitted diseases (STD) including HIV/AIDS are also spreading rapidly. However, young people lack basic information on AIDS/STD and do not know how to protect themselves from these diseases or how to avoid unintended pregnancies. Several major youth peer education programmes in China are mentioned in this paper. Among them, a four-year programme entitled the Australian-Chinese AIDS/STD/Safer Sex Peer Education Programme for Youth, is discussed in some detail. The programme has so far reached over 40000 university and school students. Evaluation results show that the programme is effective in both significantly increasing students' knowledge about AIDS/STDs and changing their attitude towards AIDS patients. In addition, the programme is highly praised by the students.


Western blot analysis of immune response to Legionella bozemanii antigens.
The immune response to individual major antigens of Legionella bozemanii was studied in 67 sera from 26 inpatients with febrile disease using immunoblotting techniques. All the patients had fever of unknown origin and showed strong serological reactions to L bozemanii that cross-reacted with Rickettsia typhi, as determined by a standard indirect microimmunofluorescence assay. Sera analysed by western blotting reacted with 12-14 molecular components of L bozemanii with apparent molecular weights ranging from 14,000 to 94,000 daltons. These reactions compared well with a reference system using antisera of rabbits immunised with the same strain of Legionella. The three major cross-reactive components of R typhi were polypeptide antigens of 94,000, 67,000 and 43,000 daltons. It is concluded that western blotting can help in the differential diagnosis of patients with fever of unknown origin whose sera cross-react to L bozemanii and R typhi.


Characterisation of Textile Embedded Electrodes for Use in a Neonatal Smart Mattress Electrocardiography System.
Heart rate monitoring is the predominant quantitative health indicator of a newborn in the delivery room. A rapid and accurate heart rate measurement is vital during the first minutes after birth. Clinical recommendations suggest that electrocardiogram (ECG) monitoring should be widely adopted in the neonatal intensive care unit to reduce infant mortality and improve long term health outcomes in births that require intervention. Novel non-contact electrocardiogram sensors can reduce the time from birth to heart rate reading as well as providing unobtrusive and continuous monitoring during intervention. In this work we report the design and development of a solution to provide high resolution, real time electrocardiogram data to the clinicians within the delivery room using non-contact electric potential sensors embedded in a neonatal intensive care unit mattress. A real-time high-resolution electrocardiogram acquisition solution based on a low power embedded system was developed and textile embedded electrodes were fabricated and characterised. Proof of concept tests were carried out on simulated and human cardiac signals, producing electrocardiograms suitable for the calculation of heart rate having an accuracy within ±1 beat per minute using a test ECG signal, ECG recordings from a human volunteer with a correlation coefficient of ~ 87% proved accurate beat to beat morphology reproduction of the waveform without morphological alterations and a time from application to heart rate display below 6 s. This provides evidence that flexible non-contact textile-based electrodes can be embedded in wearable devices for assisting births through heart rate monitoring and serves as a proof of concept for a complete neonate electrocardiogram monitoring system.


C5aR2 Regulates STING-Mediated Interferon Beta Production in Human Macrophages.
The complement system mediates diverse regulatory immunological functions. C5aR2, an enigmatic receptor for anaphylatoxin C5a, has been shown to modulate PRR-dependent pro-inflammatory cytokine secretion in human macrophages. However, the specific downstream targets and underlying molecular mechanisms are less clear. In this study, CRISPR-Cas9 was used to generate macrophage models lacking C5aR2, which were used to probe the role of C5aR2 in the context of PRR stimulation. cGAS and STING-induced IFN-β secretion was significantly increased in C5aR2 KO THP-1 cells and C5aR2-edited primary human monocyte-derived macrophages, and STING and IRF3 expression were increased, albeit not significantly, in C5aR2 KO cell lines implicating C5aR2 as a regulator of the IFN-β response to cGAS-STING pathway activation. Transcriptomic analysis by RNAseq revealed that nucleic acid sensing and antiviral signalling pathways were significantly up-regulated in C5aR2 KO THP-1 cells. Altogether, these data suggest a link between C5aR2 and nucleic acid sensing in human macrophages. With further characterisation, this relationship may yield therapeutic options in interferon-related pathologies.


Effect of bacteriophage application on Campylobacter jejuni loads in commercial broiler flocks.
Campylobacteriosis is the most frequent food-borne human enteritis. The major source for infection with Campylobacter spp. is broiler meat. Risk assessments consider the reduction of Campylobacter in primary production to be most beneficial for human health. The aim of this study was to test the efficacy of a bacteriophage application under commercial conditions which had proved to be effective in previous noncommercial studies under controlled experimental conditions. A phage cocktail for Campylobacter reduction was tested on three commercial broiler farms each with a control and an experimental group. Colonization of Campylobacter was confirmed prior to phage application in fecal samples. Subsequently, a phage cocktail was applied via drinking water in the experimental group (log10 5.8 to 7.5 PFU/bird). One day after phage application, Campylobacter counts of one experimental group were reduced under the detection limit (<50 CFU/g, P=0.0140) in fecal samples. At slaughter, a significant reduction of >log10 3.2 CFU/g cecal content compared to the control was still detected (P=0.0011). No significant reduction was observed in the experimental groups of the other trials. However, a significant drop in cecal Campylobacter counts occurred in a phage-contaminated control. These results suggest that maximum reduction of Campylobacter at the slaughterhouse might be achieved by phage application 1 to 4 days prior to slaughter.


Hepatitis B virus S mutants in liver transplant recipients who were reinfected despite hepatitis B immune globulin prophylaxis.
Long-term hepatitis B immune globulin (HBIG) has been shown to reduce hepatitis B virus (HBV) reinfection in patients transplanted for hepatitis B. The aim of this study was to determine the prevalence of HBV S gene mutations in liver transplant recipients who developed recurrent hepatitis B despite HBIG prophylaxis, and to determine if these mutations can revert after withdrawal of HBIG. The entire S gene sequences in pre- and posttransplant sera from 20 patients who developed recurrent hepatitis B despite HBIG prophylaxis were compared. Ten (50%) patients had 18 amino acid substitutions involving the 'a' determinant in the posttransplant samples. These mutations were absent in 93% of the pretransplantation clones analyzed. There was a significant correlation between the development of mutations in the 'a' determinant region and the duration of HBIG therapy. Most of the mutations result in changes in predicted antigenicity of the S protein. During follow-up, mutations in 14 (78%) of 18 affected codons in the 'a' determinant region reverted back to the pretransplantation sequences; only 1 codon had a de novo change after the withdrawal of HBIG. Two control patients who did not receive HBIG had no change in the 'a' determinant in their posttransplantation samples. These data support the hypothesis that mutations in the S gene were induced or selected by immune pressure exerted by HBIG. HBV S mutants may play a role in HBV reinfection in liver transplant recipients who received HBIG prophylaxis.


[Effect of time of day and fatigue on psychophysiological functions of an operator].
Time of the day has been found to have an important effect on the intersystemic relations of the psychophysiological functions of an operator, while the activity of the cardiovascular system (CVS) depended not only on the time of the day but also, to a large extent, on fatigue. The morning time potentiated the manifestations of staying awake but the evening time--those of fatigue. Relatively less pronounced changes in the functioning of an organism in the morning, induced by fatigue, gave evidence for both a higher level of activation of the organism and relatively higher level of the central interactions in it. Accumulated fatigue potentiated the manifestations of daily variations of the functional activity of an operator. The second subsequent night shift has been shown to have a higher initial activity of the CVS, as compared to the second day shift, and it was characterised by an increased difference between systolic and diastolic blood pressure as compared to the first night shift. Effect of overshift fatigue shows an increase in the vascular component and a decrease in the cardiac one in the regulation of blood circulation. Mental performance was shown to be less dependent on time of the day and fatigue as compared to the CVS activity. Short-term memory was more sensitive to fatigue and more often correlated with the parameters of the CVS than attention did.


Duration of indwelling drain following instrumented posterolateral fusion of the lumbar spine does not predict surgical site infection requiring reoperation.
The objective of this study was to determine the incidence and predictors of reoperation for surgical site infections (SSI) among patients whose lumbar, closed wound suction drains were removed in the inpatient setting prior to hospital discharge (pre-discharge cohort) versus after inpatient discharge during the first follow up visit (post-discharge cohort). All patients who were admitted for first-time, posterolateral decompression and fusion for degenerative lumbar spine disease were retrospectively reviewed at a single institution. In order to eliminate biases, neither the pre-discharge nor post-discharge cohorts experienced any intra-/postoperative sentinel events other than the primary outcome measure: reoperation for SSI. Of 209 patients in the pre-discharge (n=130) and post-discharge (n=79) cohorts, 15 patients required reoperation for SSI. Although time to drain discontinuation was significantly longer in the post-discharge (8.28days) than the pre-discharge (4.65days) cohorts (p<0.001), the incidences of reoperation for SSI did not significantly differ (6.33 vs 7.69%, respectively, p=0.711). In a multivariable regression, only smoking (OR=5.75, p=0.007) and depression (OR=4.11, p=0.040) predicted reoperation for SSI. Neither time to drain removal nor setting of drain removal was a predictor of reoperation for SSI. Although time to drain discontinuation was expectedly longer in the post-discharge versus pre-discharge cohorts, the incidences of reoperation for SSI did not significantly differ. Neither time to drain removal nor setting of drain removal predicted reoperation for SSI. These results suggest that patients may be safely discharged from the hospital with the surgical drain in place.


Acute hepatitis in an opossum (Didelphis virginiana) infected with Salmonella turnidorp.
Salmonella turnidorp was isolated in pure culture from the liver of an opossum that died of acute hepatitis. Microscopically, there were random foci of hepatic coagulation necrosis and gram negative bacteria within hepatocytes. The Salmonella turnidorp isolate was aberrant in that it did not utilize citrate and did not agglutinate by a commercial Salmonella polyvalent antiserum. Additional Salmonella serotypes, including Salmonella mbandaka, Salmonella rubislaw, and Salmonella anatum were isolated from three of five healthy opossums caught in Texas.


[Clinical aspects of dilated cardiomyopathy in pediatric age].
Natural history and prognostic factors of dilated cardiomyopathy in pediatric age are not well identified so far. The course of the disease, often, presents "odd" characteristics: some of the patients show clinical improvement, someone remains unchanged and some of them have a negative evolution in a very short time. It is necessary to identify the group of patients having a poor outcome in order to define prognostic factors of impending death so that a cardiac transplant could be offered to them as a reasonable therapeutic choice. Twenty-two children, 9 males and 13 females (median age 5 +/- 5 years) with dilated cardiomyopathy were studied. Their natural history was followed for a period of 40 +/- 30 months. Clinical data, laboratory studies (electrocardiogram, Holter monitoring, echocardiogram M-mode, 2-D, Doppler and chest X-ray) were taken into consideration. At the end of follow-up period 7 patients showed improvement in cardiac status (31.8%), 7 remained unchanged (31.8%) and 8 (36%) got worse; 9 died. The survival curve showed a rate of 72% at 12 months and of 59.3% at 60 months. Of 9 died patients, 2 died suddenly and seven for congestive heart failure. Clinical findings and laboratory investigations of 13 survived patients (Group I) and 9 died patients (Group II) were compared to establish the risk factors of poor outcome. The fractional shortening is the only significant factor of poor outcome (p less than 0.025). Other negative factors were identified more in Group II than in Group I.(ABSTRACT TRUNCATED AT 250 WORDS)


Various solvent systems for solubility enhancement of enrofloxacin.
Solubility enhancement of antimicrobial drug enrofloxacin has been studied using a series of co-solvents and surfactants. Aqueous solubility of enrofloxacin could be increased up to 26 times. Co-solvents alone produced only small increase in solubility. However, the combined effect of co-solvents and buffer was synergistic and a large increase in solubility could be attained. Ionic surfactants were found to be much better solubilizing agents than non-ionic surfactant. Amongst ionic surfactants, solubility was found to be very high in anionic surfactant, sodium dodecylsulphate as compared to the cationic surfactant, cetyltrimethylammonium bromide. Up to 3.8 mg/ml of enrofloxacin could be dissolved in sodium dodecylsulphate. Mechanism of solubilization has been proposed and surfactant solubilization parameters have been calculated.


Birthweight and risk of type 1 diabetes in children and young adults: a population-based register study.
We investigated the association between type 1 diabetes and birthweight by age at disease onset. This population-based case-referent study used data from two nationwide case registers that are linked to the Swedish Medical Birth Registry and cover incident cases of type 1 diabetes in the 0- to 14-year (since 1 July 1977) and 15- to 34-year age groups (since 1 January 1983). Of the cases linked to the Medical Birth Registry, a total of 9,283 cases with onset before 15 years of age was recorded before 1 January 2003, and 1,610 cases were recorded with onset before 30 years of age and born after 1973 (together 95% of eligible cases). Multiple births and babies of diabetic mothers were excluded. Sex-specific birthweight by gestational week is expressed as multiples of the standard deviation (SDS) and adjusted for year of birth, maternal age and parity. Cases with onset before 10 years of age (n = 5,792) showed a significant linear trend in odds ratio (OR) by SDS of adjusted birthweight (OR by SDS: 0.062; 95% CI: 0.037-0.086; p < 0.0001), while cases with onset at the age of 10-29 years showed no significant trend (OR by SDS: 0.004; 95% CI: -0.007 to 0.0014; p = 0.22). The association between type 1 diabetes risk and birthweight seems to be limited to cases with disease onset in younger years.


Development and Assessment of a Predictive Score for Vertebral Compression Fracture After Stereotactic Body Radiation Therapy for Spinal Metastases.
Vertebral compression fracture (VCF) is a potential adverse effect following treatment with stereotactic body radiation therapy (SBRT) for spinal metastases. To develop and assess a risk stratification model for VCF after SBRT. This retrospective cohort study conducted at a high-volume referral center included 331 patients who had undergone 464 spine SBRT treatments from December 2007 through October 2019. Data analysis was conducted from November 1, 2020, to August 17, 2021. Exclusions included proton therapy, prior surgical intervention, vertebroplasty, or missing data. One and 3 fraction spine SBRT treatments were most commonly delivered. Single-fraction treatments generally involved prescribed doses of 16 to 24 Gy (median, 20 Gy; range, 16-30 Gy) to gross disease compared with multifraction treatment that delivered a median of 30 Gy (range, 21-50 Gy). The VCF and radiography components of the spinal instability neoplastic score were determined by a radiologist. Recursive partitioning analysis was conducted using separate training (70%), internal validation (15%), and test (15%) sets. The log-rank test was the criterion for node splitting. Of the 331 participants, 88 were women (27%), and the mean (IQR) age was 63 (59-72) years. With a median follow-up of 21 months (IQR, 11-39 months), we identified 84 VCFs (18%), including 65 (77%) de novo and 19 (23%) progressive fractures. There was a median of 9 months (IQR, 3-21 months) to developing a VCF. From 15 candidate variables, 6 were identified using the backward selection method, feature importance testing, and a correlation heatmap. Four were selected via recursive partitioning analysis: epidural tumor extension, lumbar location, gross tumor volume of more than 10 cc, and a spinal instability neoplastic score of more than 6. One point was assigned to each variable, and the resulting multivariable Cox model had a concordance of 0.760. The hazard ratio per 1-point increase for VCF was 1.93 (95% CI, 1.62-2.30; P < .001). The cumulative incidence of VCF at 2 years (with death as a competing risk) was 6.7% (95% CI, 4.2%-10.7%) for low-risk (score, 0-1; 273 [58.3%]), 17.0% (95% CI, 10.8%-26.7%) for intermediate-risk (score, 2; 99 [21.3%]), and 35.4% (95% CI, 26.7%-46.9%) for high-risk cases (score, 3-4; 92 [19.8%]) (P < .001). Similar results were observed for freedom from VCF using stratification. The results of this cohort study identify a subgroup of patients with high risk for VCF following treatment with SBRT who may potentially benefit from undergoing prophylactic spinal stabilization or vertebroplasty.


Changes in social inequalities in health in the Basque Country.
To determine the extent of the inequalities in self reported health between socioeconomic groups and its changes over time in the Basque Country (Spain). Cross sectional data on the association between occupation, education and income and three health indicators was obtained from the Basque Health Surveys of 1986 and 1992. Representative population samples were analysed. In 1986 the number of respondents was 24 657 and in 1992, 13 277. Basque Country, Spain. The effect of socioeconomic position on health and the magnitude of social inequalities in health were quantified using the odds ratios based on logistic regression analysis, and the Relative Index of Inequality. As was expected, social inequalities in self reported health existed in both surveys, but the social gradient was greater in 1992. Social differences varied according to gender and health indicator. According to education an increase in social inequalities was observed consistently in all the health indicators except long term conditions in women. A consistent increase in inequalities in limiting longstanding illness was also observed according to all socioeconomic indicators. These results agree to a large extent with those of previous studies in other countries. In this context the unequal distribution of material circumstances and working conditions between socioeconomic groups seem to play a major part in health inequalities. The worsening of the labour market during this period and the onset of a new economic recession may explain the increase in social inequalities over time.


Statistical assessment of the stability of neural movement representations.
In systems neuroscience, neural activity that represents movements or sensory stimuli is often characterized by spatial tuning curves that may change in response to training, attention, altered mechanics, or the passage of time. A vital step in determining whether tuning curves change is accounting for estimation uncertainty due to measurement noise. In this study, we address the issue of tuning curve stability using methods that take uncertainty directly into account. We analyze data recorded from neurons in primary motor cortex using chronically implanted, multielectrode arrays in four monkeys performing center-out reaching. With the use of simulations, we demonstrate that under typical experimental conditions, the effect of neuronal noise on estimated preferred direction can be quite large and is affected by both the amount of data and the modulation depth of the neurons. In experimental data, we find that after taking uncertainty into account using bootstrapping techniques, the majority of neurons appears to be very stable on a timescale of minutes to hours. Lastly, we introduce adaptive filtering methods to explicitly model dynamic tuning curves. In contrast to several previous findings suggesting that tuning curves may be in constant flux, we conclude that the neural representation of limb movement is, on average, quite stable and that impressions to the contrary may be largely the result of measurement noise.


Want climate-change adaptation? Evolutionary theory can help.
The idea of adaptation, in which an organism or population becomes better suited to its environment, is used in a variety of disciplines. Originating in evolutionary biology, adaptation has been a central theme in biological anthropology and human ecology. More recently, the study of adaptation in the context of climate change has become an important topic of research in the social sciences. While there are clearly commonalities in the different uses of the concept of adaptation in these fields, there are also substantial differences. We describe these differences and suggest that the study of climate-change adaptation could benefit from a re-integration with biological and evolutionary conceptions of human adaptation. This integration would allow us to employ the substantial theoretical tools of evolutionary biology and anthropology to understand what promotes or impedes adaptation. The evolutionary perspective on adaptation focuses on diversity because diversity drives adaptive evolution. Population structures are also critical in facilitating or preventing adaptation to local environmental conditions. This suggests that climate-change adaptation should focus on the sources of innovation and social structures that nurture innovations and allow them to spread. Truly innovative ideas are likely to arise on the periphery of cohesive social groups and spread inward. The evolutionary perspective also suggests that we pay careful attention to correlated traits, which can distort adaptive trajectories, as well as to the importance of risk management in adaptations to variable or uncertain environments. Finally, we suggest that climate-change adaptation could benefit from a broader study of how local groups adapt to their dynamic environments, a process we call "autochthonous adaptation."


Fourier-transform resonance shear measurement for studying confined liquids.
The resonance shear measurement we recently developed is an efficient tool for investigating the rheological and tribological properties of liquid nanofilms confined between solid surfaces with varying film thicknesses. However, the previously employed resonance shear measurement measured shear responses at various oscillation frequencies for one film thickness (the frequency scanning method), so it required at least several minutes to obtain a reliable resonance curve. The fast Fourier-transform resonance shear method was developed to rapidly acquire the resonance curves. The obtained curves were very similar to those obtained by the frequency scanning method. The time necessary for the FFT method was 2-10 s, much shorter than the time required for the scanning method. This technique is easy to use and extends the applicability of the method to volatile liquids and unstable samples.


Endurance exercise with and without a thermal clamp: effects on leukocytes and leukocyte subsets.
To test how leukocyte responses to endurance exercise were modified by clamping body temperature, nine men (27.3 +/- 6.0 yr) completed four 80-min immersions to midchest at water temperatures of 23 or 39 degrees C; two tests included 40-min of cycle ergometer exercise at 65% of aerobic power. When the subjects were exercising, rectal temperature peaked at 39.1 +/- 0.4 degrees C in the warm water and 37.8 +/- 0.3 degrees C in the cool water. When the subjects were sitting in warm water, rectal temperature closely matched the core temperature during exercise in cool water, whereas when they were sitting in cool water, rectal temperatures decreased to 36.4 +/- 0.6 degrees C. Total and differential white cell counts were determined by using a Coulter counter, and cortisol and growth hormone concentrations were determined by radioimmunoassay; all data were adjusted for changes of blood and plasma volumes. Heat clamping during exercise substantially reduced the rise in white cell, lymphocyte, and granulocyte counts but not the increase in monocyte count. Clamping also abolished previously observed associations between cell counts and cortisol and weakened associations with growth hormone concentrations (D. A. McCarthy and M. M. Dale. Sports Med. 6: 333-363, 1988). We conclude that both exercise and a rise of core temperature contribute to the changes in white cell and subset counts during and immediately after moderate exercise. Both cortisol and growth hormone concentrations appear to be mediators of these responses.


Integration of Preferences in Decomposition Multiobjective Optimization.
Rather than a whole Pareto-optimal front, which demands too many points (especially in a high-dimensional space), the decision maker (DM) may only be interested in a partial region, called the region of interest (ROI). In this case, solutions outside this region can be noisy to the decision-making procedure. Even worse, there is no guarantee that we can find the preferred solutions when tackling problems with complicated properties or many objectives. In this paper, we develop a systematic way to incorporate the DM's preference information into the decomposition-based evolutionary multiobjective optimization methods. Generally speaking, our basic idea is a nonuniform mapping scheme by which the originally evenly distributed reference points on a canonical simplex can be mapped to new positions close to the aspiration-level vector supplied by the DM. By this means, we are able to steer the search process toward the ROI either directly or interactively and also handle many objectives. Meanwhile, solutions lying on the boundary can be approximated as well given the DM's requirements. Furthermore, the extent of the ROI is intuitively understandable and controllable in a closed form. Extensive experiments on a variety of benchmark problems with 2 to 10 objectives, fully demonstrate the effectiveness of our proposed method for approximating the preferred solutions in the ROI.


Catheter-directed therapy for acute pulmonary embolism in children.
Acute pulmonary embolism is a life-threatening condition and rarely occurs in children. In adults, catheter-directed therapy emerges as a potentially safer and effective therapeutic option. However, there is a paucity of data on the safety and efficacy of catheter-directed therapy for pulmonary embolism in children. We report a single-centred experience of catheter-directed therapy for acute pulmonary embolism in children. This is a retrospective study of children who had no CHD and underwent catheter-directed therapy at Detroit Medical Center during a 12-year period from 2005 to 2017. Demographic and clinical data associated with pulmonary embolism were collected along with the outcome. A total of nine patients of median age 16 years with the range from 12 to 20 received catheter-directed therapy for sub-massive (n = 6) and massive pulmonary embolism (n = 3). Among nine patients, one patient received Angiojet thrombectomy and balloon angioplasty, whereas eight patients received catheter-directed thrombolysis using tissue plasminogen activator through infusion catheters (n = 3) or EkoSonic ultrasound-accelerated thrombolysis system (n = 5). In four out of five patients treated with EkoSonic, significant clinical improvement was noticed within 24 hours. Among seven patients who survived, two patients had minor gastrointestinal bleeding with median hospital stay of 8 days with the range from 5 to 24 days, and two patients with massive pulmonary embolism died possibly due to delayed institution of catheter-directed therapy. Catheter-directed therapy with/without EkoSonic is an emerging alternative therapy for sub-massive and massive pulmonary embolism in children. A timely institution of catheter-directed therapy appeared important to improve the outcome.


Pharmacokinetics of ε-Aminocaproic Acid in Neonates Undergoing Cardiac Surgery with Cardiopulmonary Bypass.
Antifibrinolytic medications such as ε-aminocaproic acid (EACA) are used in pediatric heart surgery to decrease surgical bleeding and transfusion. Dosing schemes for neonates are often based on adult regimens, or are simply empiric, in part due to the lack of neonatal pharmacokinetic information. The authors sought to determine the pharmacokinetics of EACA in neonates undergoing cardiac surgery and to devise a dosing regimen for this population. Ten neonates undergoing cardiac surgery with cardiopulmonary bypass were given EACA according to standard practice, and blood was drawn at 10 time points to determine drug concentrations. Time-concentration profiles were analyzed using nonlinear mixed effects models. Parameter estimates (standardized to a 70-kg person) were used to develop a dosing regimen intended to maintain a target concentration shown to inhibit fibrinolysis in neonatal plasma (50 mg/l). Pharmacokinetics were described using a two-compartment model plus an additional compartment for the cardiopulmonary bypass pump. First-order elimination was described with a clearance of 5.07 l/h × (WT/70). Simulation showed a dosing regimen with a loading dose of 40 mg/kg and an infusion of 30 mg · kg · h, with a pump prime concentration of 100 mg/l maintained plasma concentrations above 50 mg/l in 90% of neonates during cardiopulmonary bypass surgery. EACA clearance, expressed using allometry, is reduced in neonates compared with older children and adults. Loading dose and infusion dose are approximately half those required in children and adults.


Integrating data and knowledge to identify functional modules of genes: a multilayer approach.
Characterizing the modular structure of cellular network is an important way to identify novel genes for targeted therapeutics. This is made possible by the rising of high-throughput technology. Unfortunately, computational methods to identify functional modules were limited by the data quality issues of high-throughput techniques. This study aims to integrate knowledge extracted from literature to further improve the accuracy of functional module identification. Our new model and algorithm were applied to both yeast and human interactomes. Predicted functional modules have covered over 90% of the proteins in both organisms, while maintaining a comparable overall accuracy. We found that the combination of both mRNA expression information and biomedical knowledge greatly improved the performance of functional module identification, which is better than those only using protein interaction network weighted with transcriptomic data, literature knowledge, or simply unweighted protein interaction network. Our new algorithm also achieved better performance when comparing with some other well-known methods, especially in terms of the positive predictive value (PPV), which indicated the confidence of novel discovery. Higher PPV with the multiplex approach suggested that information from both sources has been effectively integrated to reduce false positive. With protein coverage higher than 90%, our algorithm is able to generate more novel biological hypothesis with higher confidence.


Chickenpox in adults - clinical management.
Acute varicella zoster virus (VZV) infection, or chickenpox, is still perceived by many as a mild infection of childhood. However, chickenpox is increasingly common in adults and adolescents who together with immunosuppressed individuals are at a higher risk of severe infection. Antiviral therapy is available which both ameliorates symptoms and decreases the severity of chickenpox if administered early in the course of the infection. Passive immunisation with varicella zoster immunoglobulin (VZIG) may prevent or attenuate infection following exposure to varicella of an immunocompromised or pregnant individual or a neonate. Active immunisation is available and is universal in many developed countries. This review reflects current best practice in management of chickenpox in adults by specialist physicians in the UK. The accompanying flowchart has been formulated to guide emergency physicians and general practitioners through the decision-making process regarding treatment and admission for specialist care.


Time, action and psychosis: using subjective time to investigate the effects of ketamine on sense of agency.
Sense of agency refers to the experience of initiating and controlling actions in order to influence events in the outside world. A disturbed sense of agency is found in certain psychiatric and neurological disorders, most notably schizophrenia. Sense of agency is associated with a subjective compression of time: actions and their outcomes are perceived as bound together in time. This is known as 'intentional binding' and, in healthy adults, depends partly on advance prediction of action outcomes. Notably, this predictive contribution is disrupted in patients with schizophrenia. In the present study we aimed to characterise the psychotomimetic effect of ketamine, a drug model for psychosis, on the predictive contribution to intentional binding. It was shown that ketamine produced a disruption that closely resembled previous data from patients in the early, prodromal, stage of schizophrenic illness. These results are discussed in terms of established models of delusion formation in schizophrenia. The link between time and agency, more generally, is also considered.


The immunotherapy candidate TNFSF4 may help the induction of a promising immunological response in breast carcinomas.
Immune checkpoint blockade, an immunotherapy, has been applied in multiple systemic malignancies and has improved overall survival to a relatively great extent; whether it can be applied in breast cancer remains unknown. We endeavored to explore possible factors that may influence immunotherapy outcomes in breast cancer using several public databases. The possible treatment target TNF superfamily member 4 (TNFSF4) was selected from many candidates based on its abnormal expression profile, survival-associated status, and ability to predict immune system reactions. For the first time, we identified the oncogenic features of TNFSF4 in breast carcinoma. TNFSF4 was revealed to be closely related to treatment that induced antitumor immunity and to interact with multiple immune effector molecules and T cell signatures, which was independent of endocrine status and has not been reported previously. Moreover, the potential immunotherapeutic approach of TNFSF4 blockade showed underlying effects on stem cell expansion, which more strongly and specifically demonstrated the potential effects of applying TNFSF4 blockade-based immunotherapies in breast carcinomas. We identified potential targets that may contribute to breast cancer therapies through clinical analysis and real-world review and provided one potential but crucial tool for treating breast carcinoma that showed effects across subtypes and long-term effectiveness.


The Complement System in the Pathophysiology of Pregnancy and in Systemic Autoimmune Rheumatic Diseases During Pregnancy.
The complement system plays a double role in pregnancy exerting both protective and damaging effects at placental level. Complement activation at fetal-maternal interface participates in protection against infectious agents and helps remove apoptotic and necrotic cells. Locally synthesized C1q contributes to the physiologic vascular remodeling of spiral arteries characterized by loss of smooth muscle cells and transformation into large dilated vessels. Complement activation triggered by the inflammatory process induced by embryo implantation can damage trophoblast and other decidual cells that may lead to pregnancy complications if the cells are not protected by the complement regulators CD55, CD46, and CD59 expressed on cell surface. However, uncontrolled complement activation induces placental alterations resulting in adverse pregnancy outcomes. This may occur in pathological conditions characterized by placental localization of complement fixing antibodies directed against beta2-glycoprotein 1, as in patients with anti-phospholipid syndrome, or circulating immune complexes deposited in placenta, as in patients with systemic lupus erythematosus. In other diseases, such as preeclampsia, the mechanism of complement activation responsible for complement deposits in placenta is unclear. Conflicting results have been reported on the relevance of complement assays as diagnostic and prognostic tools to assess complement involvement in pregnant patients with these disorders.


Conservation of resources: indications for intensive care monitoring after upper airway surgery on patients with obstructive sleep apnea.
Although significant complications can result after upper airway surgery for obstructive sleep apnea (OSA), there is a lack of consensus regarding the most appropriate level of monitoring in the perioperative period. A retrospective analysis was performed on the operative records of 109 adult patients who underwent 125 surgical procedures from January 1, 1991, to May 31, 1996, with particular emphasis on complications that would have mandated intensive care monitoring and management. Airway complications occurred in one patient (0.8%), who became obstructed immediately after surgery; he responded to naloxone and suctioning. Five other patients (4%) suffered oxygen desaturation to levels below 90% (none fell below 80%, and in only one case was it below the lowest preoperative oxygen saturation level). Cardiac complications, primarily significant hypertension, were the most common adverse events. Four (3.2%) bleeding complications were encountered; all occurred after discharge from the hospital. Routine postoperative intensive care monitoring for all adult patients undergoing upper airway surgery for OSA is unnecessary. Although high-risk patients cannot always be identified preoperatively, significant complications generally emerge within 2 hours after surgery. Therefore a decision regarding the level of postoperative monitoring needed may be made with confidence during the period of time that the patient is in the recovery room.


Strictly Lobar Cerebral Microbleeds Are Associated With Cognitive Impairment.
Different distributions of cerebral microbleeds (CMBs) are associated with distinct pathological mechanisms. Lobar CMBs are thought to be related to cerebral amyloid angiopathy, whereas deep or infratentorial CMBs are related to hypertensive vasculopathy. The present study aimed to evaluate the effects of CMBs and their locations on a variety of cognitive domains. Study subjects were selected from the community-based I-Lan Longitudinal Aging Study. We assessed cognitive domains, including verbal memory, language, visuospatial executive function, and verbal executive function. CMBs were evaluated using 3T susceptibility-weighted magnetic resonance imaging. We studied 959 subjects (mean±SD, 62.5±8.6 years; 425 [44.3%] men). CMBs were found in 14.2% of the population. We classified subjects with CMBs into 2 different groups based on the locations of their CMBs: (1) deep or infratentorial (85 subjects, 8.8% of population) and (2) strictly lobar (49, 5.1%). Multivariate linear analysis showed that strictly lobar CMBs were significantly associated with deficits in global cognitive function (Mini-Mental State Examination) and visuospatial executive function, as determined by the copy test of the Taylor complex figure test and the clock drawing test. We adjusted our results for age, sex, years of education, cardiovascular risk factors, and other markers of cerebral small vessel disease, lacunes, and white matter hyperintensity. Deep or infratentorial CMBs were not associated with changes in cognitive function in our population. Strictly lobar, but not deep or infratentorial, CMBs are associated with changes in cognitive function, especially in visuospatial executive functions. Cerebral amyloid angiopathy may be the underlying pathology associated with CMB-related cognitive impairment.


Effects of feline leukemia virus infection on neutrophil chemotaxis in vitro.
A procedure for measuring in vitro feline neutrophil chemotaxis was developed, using a modified Boyden chamber apparatus and 3-microns-pore polycarbonate filters. A pooled feline serum sample was used as the chemoattractant. Chemotaxis was evaluated in 5 groups of cats: group 1-specific-pathogen-free cats that had not been exposed to feline leukemia virus (FeLV); group 2-previremic, FeLV-infected, specific-pathogen-free cats; group 3-FeLV-viremic, subclinically affected cats; group 4-FeLV-viremic, clinically affected cats; and group 5-sick cats that were not infected with FeLV. Neutrophils from the viremic, clinically affected cats had significantly lower (P less than 0.025) chemotactic responses than did those from subclinically affected, viremic cats. Conversely, neutrophils from cats that were ill due to causes other than FeLV had the highest mean chemotactic values. Among the viremic, subclinically affected cats, a linear relationship was found between age and chemotaxis, indicating that impairment of neutrophil function may be greater in younger viremic cats. However, FeLV-infected cats can not be identified on the basis of neutrophil chemotaxis.


The interactive effects of competition and predation risk on dispersal in an insect.
Dispersal dynamics have significant consequences for ecological and evolutionary processes. Previous work has demonstrated that dispersal can be context-dependent. However, factors affecting dispersal are typically considered in isolation, despite the probability that individuals make dispersal decisions in response to multiple, possibly interacting factors. We examined whether two ecological factors, predation risk and intraspecific competition, have interactive effects on dispersal dynamics. We performed a factorial experiment in mesocosms using backswimmers (Notonecta undulata), flight-capable, semi-aquatic insects. Emigration rates increased with density, and increased with predation risk at intermediate densities; however, predation had minimal effects on emigration at high and low densities. Our results indicate that factorial experiments may be required to understand dispersal dynamics under realistic ecological conditions.


[Correlation between 99m Tc-tetrofosmin uptake and P-glycoprotein expression in non-small-cell lung cancer].
We used thoracic SPECT to study the 99mTc-tetrofosmin (TF) uptake in patients with non-small-cell-lung-carcinoma (NSCLC). The results were compared with the percentage of P-glycoprotein (Pgp) found in flow cytometric analysis (FC) of samples of surgically-resected tumor tissue. A total of 21 patients with NSCLC were studied by means 99mTC-TF and thoracic SPECT. Image analysis included the determination of the TF uptake rate in the lung mass with respect to that of healthy tissue of the contralateral lung. These rates were compared with the percentage of Pgp expression according to FC. FC analysis was also carried out in 16 samples of healthy lung tissue obtained from the patients. In healthy lung tissue, the mean Pgp expression according to FC was 4.58 +/- 1.87%. The cutoff value used to differentiate between Pgp positive and Pgp negative tumors was considered to be the mean plus two standard deviations (8.32). The Pgp-positive tumors (> 8.32%) presented significantly lower uptake levels (1.28 +/- 0.39) than the Pgp-negative lesions (1.66 +/- 0.33) (p = 0.029). There is a inverse correlation between the Pgp expression as determined by FC analysis and 99mTc-TF in NSCLC. Thus, this radiopharmaceutical provides rapid and non-invasive information on Pgp expression in these lesions.


Problems with Centrality Measures in Psychopathology Symptom Networks: Why Network Psychometrics Cannot Escape Psychometric Theory.
Understanding patterns of symptom co-occurrence is one of the most difficult challenges in psychopathology research. Do symptoms co-occur because of a latent factor, or might they directly and causally influence one another? Motivated by such questions, there has been a surge of interest in network analyses that emphasize the putatively direct role symptoms play in influencing each other. In this critical paper, we highlight conceptual and statistical problems with using centrality measures in cross-sectional networks. In particular, common network analyses assume that there are no unmodeled latent variables that confound symptom co-occurrence. The traditions of clinical taxonomy and test development in psychometric theory, however, greatly increase the possibility that latent variables exist in symptom data. In simulations that include latent variables, we demonstrate that closeness and betweenness are vulnerable to spurious covariance among symptoms that connect subgraphs (e.g., diagnoses). We further show that strength is redundant with factor loading in several cases. Finally, if a symptom reflects multiple latent causes, centrality metrics reflect a weighted combination, undermining their interpretability in empirical data. Our results suggest that it is essential for network psychometric approaches to examine the evidence for latent variables prior to analyzing or interpreting patterns at the symptom level. Failing to do so risks identifying spurious relationships or failing to detect causally important effects. Altogether, we argue that centrality measures do not provide solid ground for understanding the structure of psychopathology when latent confounding exists.


The ABC transporters in Candidatus Liberibacter asiaticus.
Candidatus Liberibacter asiaticus (Ca. L. asiaticus) is a Gram-negative bacterium and the pathogen of Citrus Greening disease (Huanglongbing, HLB). As a parasitic bacterium, Ca. L. asiaticus harbors ABC transporters that play important roles in exchanging chemical compounds between Ca. L. asiaticus and its host. Here, we analyzed all the ABC transporter-related proteins in Ca. L. asiaticus. We identified 14 ABC transporter systems and predicted their structures and substrate specificities. In-depth sequence and structure analysis including multiple sequence alignment, phylogenetic tree reconstruction, and structure comparison further support their function predictions. Our study shows that this bacterium could use these ABC transporters to import metabolites (amino acids and phosphates) and enzyme cofactors (choline, thiamine, iron, manganese, and zinc), resist to organic solvent, heavy metal, and lipid-like drugs, maintain the composition of the outer membrane (OM), and secrete virulence factors. Although the features of most ABC systems could be deduced from the abundant experimental data on their orthologs, we reported several novel observations within ABC system proteins. Moreover, we identified seven nontransport ABC systems that are likely involved in virulence gene expression regulation, transposon excision regulation, and DNA repair. Our analysis reveals several candidates for further studies to understand and control the disease, including the type I virulence factor secretion system and its substrate that are likely related to Ca. L. asiaticus pathogenicity and the ABC transporter systems responsible for bacterial OM biosynthesis that are good drug targets.


A 6-month follow-up study of posttraumatic stress and anxiety/depressive symptoms in Korean children after direct or indirect exposure to a single incident of trauma.
The aims of this study were to examine the symptoms of posttraumatic stress and anxiety/depression in Korean children after direct or indirect exposure to a single incident of trauma during a fire-escape drill and to assess the incidence of psychiatric disorders in this population. A total of 1,394 students who attended the elementary school at which the traumatic event took place were evaluated using self-administered questionnaires (the Child Posttraumatic Stress Disorder-Reaction Index [CPTSD-RI], State Anxiety Scale of the State-Trait Anxiety Inventory for Children [STAIC], and Children's Depression Inventory [CDI]), as well as structured diagnostic interviews (Diagnostic Interview Schedule for Children, Version-IV [DISC-IV]) at 2 days (time point 1), 2 months (time point 2), and 6 months (time point 3) after the incident. The 335 students who witnessed the accident were defined as the direct-exposure group, and the remaining students (n = 1,059) were defined as the indirect-exposure group. The study was conducted from May to November 2007. At time point 1, the prevalence of severe posttraumatic stress disorder (PTSD), anxiety, and depressive symptoms was 18.2%, 5.5%, and 3.4%, respectively. The prevalence of severe PTSD symptoms, as measured by the CPTSD-RI, was significantly higher in the direct-exposure group than in the indirect-exposure group (36.6% vs 12.7%, respectively; P < .001). At time point 2, the prevalence of severe PTSD symptoms was 7.4% (14.0% in the direct-exposure group and 4.9% in the indirect-exposure group, P < .001). The mean total CPTSD-RI score was significantly higher (P < .001) in the direct-exposure group than in the indirect-exposure group. At time point 3, thirty-eight of the 58 subjects (65.5%) evaluated with the DISC-IV in the direct-exposure group had 1 or more of the 7 anxiety/depressive disorders assessed, including subthreshold diagnoses. Among the diagnoses meeting full DSM-IV criteria for each disorder, agoraphobia was the most prevalent (22.4%), followed by generalized anxiety disorder (13.8%), separation anxiety disorder (6.9%), PTSD (5.2%), and social phobia (5.2%). When the subthreshold diagnoses were considered along with the full syndrome diagnoses, separation anxiety disorder was the most common diagnosis (41.4%), followed by agoraphobia (34.5%), obsessive-compulsive disorder (22.4%), PTSD (20.7%), and social phobia (20.7%). The results of this study provide important evidence that various anxiety/depressive disorders, in addition to PTSD, might follow after direct or indirect exposure to trauma. Our findings highlight the importance of comprehensive screening for psychiatric problems in children exposed to trauma of any scale.


Fate of the hippocampal mossy fiber projection after destruction of its postsynaptic targets with intraventricular kainic acid.
Intraventricular injections of kainic acid were used to create a model of selective cell death in order to study the fate of afferent projections that are deprived of their postsynaptic targets. This treatment rapidly destroyed hippocampal CA3 pyramidal cells, but not those neurons that give rise to their mossy fiber and entorhinal afferents. Light microscopic studies with the Timm's sulfide silver stain indicated that half or more of the mossy fiber boutons in area CA3b were lost within the first 1-3 days after kainic acid administration. This finding was confirmed by electron microscopy. Electron-dense, usually vacuolated mossy fiber boutons accounted for about 10-20% of the total population present at a 4-hour survival time, but were not encountered in control rats nor at survival times longer than 1 day. Other mossy fiber boutons remained electron lucent, but enlarged, became more rounded in shape, and suffered an apparent loss of synaptic vesicles. It is suggested that degeneration of some mossy fiber boutons and resorption of others into the axon may have accounted for the precipitous decline in their number. The dendritic excrescences contacted by these boutons were nearly all undergoing electron-dense degeneration 4 hours after kainic acid administration. In rats that survived 6-8 weeks mossy fiber boutons remained somewhat scarce, individual boutons appeared relatively small, and only one-third the normal percentage were observed to be engaged in more than one synaptic contact within a single cross section. A qualitative electron microscopic study of the entorhinal projection to area CA3 suggested a response to kainic acid treatment similar to that of the mossy fiber projection, except that no entorhinal boutons were seen to become electron dense. These findings suggest that presynaptic fibers in the mature hippocampus adjust the size of their terminal arborizations and number of synaptic contacts to accommodate a reduction in the target cell population. The rapid loss of mossy fiber boutons may be attributable to an unusual fragility of these structures when they are deprived of the mechanical support normally provided by the pyramidal cell. Finally, the ability of kainic acid administration to alter the number and distribution of presynaptic elements must be taken into account whenever this toxin is used to make selective lesions of postsynaptic cells.


Lymphatic territories (lymphosomes) in a canine: an animal model for investigation of postoperative lymphatic alterations.
Lymph node dissection is often performed as a part of surgical treatment for breast cancer and malignant melanoma to prevent malignant cells from traveling via the lymphatic system. Currently little is known about postoperative lymphatic drainage pattern alterations. This knowledge may be useful for management of recurrent cancer and prevention of breast cancer related lymphedema. We mapped the complete superficial lymphatic system of a dog and used this canine model to perform preliminary studies of lymphatic architectural changes in postoperative condition. Lymphatic territories (lymphosomes) were mapped with 4 female mongrel carcasses using an indocyanine green (ICG) fluorescent lymphography and a radiographic microinjection technique. Two live dogs were then subjected to unilateral lymph node dissection of lymph basins of the forelimb, and ICG lymphography and lymphangiogram were performed 6 months after the surgery to investigate lymphatic changes. Lymphatic patterns in the carcass were then compared with postoperative lymphatic patterns in the live dogs. Ten lymphosomes were identified, corresponding with ten lymphatic basins. Postoperative fluorescent lymphographic images and lymphangiograms in the live dogs revealed small caliber lymphatic network fulfilling gaps in the surgical area and collateral lymphatic vessels arising from the network connecting to lymph nodes in the contralateral and ipsilateral neck in one dog and the ipsilateral subclavicular vein in another dog. Our canine lymphosome map allowed us to observe lymphatic collateral formations after lymph node dissection in live dogs. This canine model may help clarify our understanding of postoperative lymphatic changes in humans in future studies.


Risk of intra-articular injection with longitudinal ultrasound-guided injection of collateral ligaments of the equine distal interphalangeal joint.
Desmopathy of the collateral ligaments of the distal interphalangeal joint is a common cause of equine foot lameness and carries a poor prognosis with conservative management. Intralesional injections may improve healing, although accuracy of radiographically guided injections is significantly less than when guided by MRI, which requires special needles. Longitudinal ultrasound-guided injection of the distal collateral ligament has not been evaluated objectively. In this prospective, anatomic study, seven equine cadaver limbs (14 collateral ligaments) were injected with methylene blue dye and radiographic contrast medium using ultrasound to guide the needle longitudinally into the collateral ligaments until contacting bone. The insertion site of the needle proximal to the coronary band was measured on the limb and the needles left in place for radiography and CT to evaluate the needle angulation, location of the contrast medium, and whether the contrast entered the distal interphalangeal joint. The limbs were frozen and sectioned with a band saw to identify the location of the dye. Fifty percentage of injections were in or around the collateral ligaments. However, the percentage of "successful" injections, defined as in the collateral ligament but not in the joint, was only 36%. All legs had dye and contrast in the joint after both ligaments had been injected. There were no significant differences between the needle angle and entry site for "successful" and "unsuccessful" injections. Findings from this study indicates that the success rate is low for injecting the distal portions of the distal interphalangeal joint collateral ligaments using ultrasound guidance alone.


The importance of iron supply during repetitive harvesting of Aster tripolium.
Aster tripolium L. is a salt marsh halophyte that has recently gained interest as a cash crop vegetable. Leaf yield and quality were investigated in plants grown with salinity in experiments with Perlite in pots and in plots on dune sand. Plants were repetitively harvested in a 14-day cycle. A. tripolium irrigated with 50mM NaCl exhibited the highest yield when grown in pots, whereas in the plot experiment no significant differences in biomass accumulation occurred up to 80mM NaCl in the irrigation water. Chemical leaf composition changed with salinity, exhibiting higher levels of electrical conductivity, total soluble solutes and the non-enzymatic antioxidant compounds ascorbic acid and polyphenols compared with control plants grown without NaCl supplementation. Using the repetitive harvest regime, leaf chlorosis occurred, a symptom shared by deficiencies in either nitrogen or iron. Comparative applications of five iron chelate formulations in plants grown with 50mM NaCl in pots revealed improved leaf colour and chlorophyll content for only two of the applied Fe-chelates. Concomitantly with leaf colour restoration, the activity of nitrate reductase, the first enzyme during nitrate assimilation, which requires heme-iron for its proper function, increased 3-fold as a result of the iron treatment in the plot experiment. Importantly, the enhancement of nitrate reductase activity was associated with a considerable decrease in the leaf nitrate concentration. Therefore, we concluded that iron deficiency, in addition to leaf chlorosis, reduces A. tripolium leaf quality as a vegetable by increasing the leaf nitrate content. Furthermore, nitrate reductase (NR) activity levels in A. tripolium leaves may act as an indicator of iron deficiency that manifests itself as reduced nitrate content owing to the higher NR activity upon proper iron nutrition. These results demonstrate the importance of salinity level and the application of an appropriate iron-chelating formulation to generate marketable yields of Aster tripolium leafy vegetable when grown commercially on dune sand.


Atezolizumab in Combination With Carboplatin and Survival Outcomes in Patients With Metastatic Triple-Negative Breast Cancer: The TBCRC 043 Phase 2 Randomized Clinical Trial.
Agents targeting programmed death ligand 1 (PD-L1) have demonstrated efficacy in triple-negative breast cancer (TNBC) when combined with chemotherapy and are now the standard of care in patients with PD-L1-positive metastatic disease. In contrast to microtubule-targeting agents, the effect of combining platinum compounds with programmed cell death 1 (PD-1)/PD-L1 immunotherapy has not been extensively determined. To evaluate the efficacy of atezolizumab with carboplatin in patients with metastatic TNBC. This phase 2 randomized clinical trial was conducted in 6 centers from August 2017 to June 2021. Patients with metastatic TNBC were randomized to receive carboplatin area under the curve (AUC) 6 alone or with atezolizumab, 1200 mg, every 3 weeks until disease progression or unacceptable toxic effects with a 3-year duration of follow-up. The primary end point was investigator-assessed progression-free survival (PFS). Secondary end points included overall response rate (ORR), clinical benefit rate (CBR), and overall survival (OS). Other objectives included correlation of response with tumor PD-L1 levels, tumor-infiltrating lymphocytes (TILs), tumor DNA- and RNA-sequenced biomarkers, TNBC subtyping, and multiplex analyses of immune markers. All 106 patients with metastatic TNBC who were enrolled were female with a mean (range) age of 55 (27-79) years, of which 12 (19%) identified as African American/Black, 1 (1%) as Asian, 73 (69%) as White, and 11 (10%) as unknown. Patients were randomized and received either carboplatin (n = 50) or carboplatin and atezolizumab (n = 56). The combination improved PFS (hazard ratio [HR], 0.66; 95% CI, 0.44-1.01; P = .05) from a median of 2.2 to 4.1 months, increased ORR from 8.0% (95% CI, 3.2%-18.8%) to 30.4% (95% CI, 19.9%-43.3%), increased CBR at 6 months from 18.0% (95% CI, 9.8%-30.1%) to 37.5% (95% CI, 26.0%-50.6%), and improved OS (HR, 0.60; 95% CI, 0.37-0.96; P = .03) from a median of 8.6 to 12.6 months. Subgroup analysis showed PD-L1-positive tumors did not benefit more from adding atezolizumab (HR, 0.62; 95% CI, 0.23-1.65; P = .35). Patients with high TILs (HR, 0.12; 95% CI, 0.30-0.50), high mutation burden (HR, 0.50; 95% CI, 0.23-1.06), and prior chemotherapy (HR, 0.59; 95% CI, 0.36-0.95) received greater benefit on the combination. Patients with obesity and patients with more than 125 mg/dL on-treatment blood glucose levels were associated with better PFS (HR, 0.35; 95% CI, 0.10-1.80) on the combination. TNBC subtypes benefited from adding atezolizumab, except the luminal androgen receptor subtype. In this randomized clinical trial, the addition of atezolizumab to carboplatin significantly improved survival of patients with metastatic TNBC regardless of PD-L1 status. Further, lower risk of disease progression was associated with increased TILs, higher mutation burden, obesity, and uncontrolled blood glucose levels. ClinicalTrials.gov Identifier: NCT03206203.


Multiple influences of blood flow on cardiomyocyte hypertrophy in the embryonic zebrafish heart.
Cardiomyocyte hypertrophy is a complex cellular behavior involving coordination of cell size expansion and myofibril content increase. Here, we investigate the contribution of cardiomyocyte hypertrophy to cardiac chamber emergence, the process during which the primitive heart tube transforms into morphologically distinct chambers and increases its contractile strength. Focusing on the emergence of the zebrafish ventricle, we observed trends toward increased cell surface area and myofibril content. To examine the extent to which these trends reflect coordinated hypertrophy of individual ventricular cardiomyocytes, we developed a method for tracking cell surface area changes and myofibril dynamics in live embryos. Our data reveal a previously unappreciated heterogeneity of ventricular cardiomyocyte behavior during chamber emergence: although cardiomyocyte hypertrophy was prevalent, many cells did not increase their surface area or myofibril content during the observed timeframe. Despite the heterogeneity of cell behavior, we often found hypertrophic cells neighboring each other. Next, we examined the impact of blood flow on the regulation of cardiomyocyte behavior during this phase of development. When blood flow through the ventricle was reduced, cell surface area expansion and myofibril content increase were both dampened, and the behavior of neighboring cells did not seem coordinated. Together, our studies suggest a model in which hemodynamic forces have multiple influences on cardiac chamber emergence: promoting both cardiomyocyte enlargement and myofibril maturation, enhancing the extent of cardiomyocyte hypertrophy, and facilitating the coordination of neighboring cell behaviors.


Hypertension and arterial disease.
There is abundant evidence that the height of an individual's systemic arterial pressure is related, in a graded way, to his chances of developing ischaemic heart disease or cerebrovascular disease. The way in which the link operates in respect of coronary disease is not yet known, and further evidence is required before we can accept that the hypertension is necessarily a direct causal factor rather than a marker of some feature in the individual's make-up which is itself linked to the onset of arterial thrombosis (the ;series' or ;parallel' wiring dilemma). In respect of haemorrhagic stroke not only have we a link but we also have a mechanism of action plus evidence that pressure reduction can modify the risk. We have a ;series' wired circuit, and the problem now facing us is to acquire the wisdom which will allow us to decide when to seek out hypertension and what we should do when we find it.