Synthetic hydrophobic peptides are substrates for P-glycoprotein and stimulate drug transport.
P-Glycoprotein functions as an ATP-driven active efflux pump for many natural products and chemotherapeutic drugs. Hydrophobic peptides have been shown to block drug uptake by P-glycoprotein, indicating that they might be transport substrates. The present study examines the interaction of the synthetic peptide series NAc-LnY-amide with the multidrug transporter. Several peptides in this series caused up to 3.5-fold enhancement of colchicine accumulation in membrane vesicles from multidrug resistant (MDR) cells, which suggests the existence of novel interactions between the binding sites for peptides and drug. Peptides did not stimulate vinblastine transport, which was inhibited as expected for competing substrates. These peptides displayed modest stimulatory effects on the ATPase activity of P-glycoprotein. None blocked azidopine photoaffinity labelling, showing that they probably occupy a binding site separate from that for the drug. Studies with 125I-labelled NAc-LLY-amide showed that it was transported by P-glycoprotein in both membrane vesicles and reconstituted proteoliposomes. Uptake of the peptide was rapid, saturable, osmotically sensitive and occurred against a concentration gradient. The enhancing effect of NAc-LLY-amide on colchicine transport was reciprocated, i.e. colchicine greatly increased the transport of labelled peptide by P-glycoprotein. Peptide transport was also modulated, both positively and negatively, by other MDR spectrum drugs. It is concluded that linear hydrophobic peptides are indeed transported by P-glycoprotein, and some have interactions with drug substrates that result in mutual stimulation of transport.


Evaluation of angiogenesis with the expression of VEGF and CD34 in human non-small cell lung cancer.
Angiogenesis is an essential process in the progression of malignant tumors and the most potent angiogenic factor is the vascular endothelial growth factor (VEGF). On the other hand, the CD34 is an endothelial antigen that has been used to highlight the microvasculature vessel density (MVD) as a direct marker of the degree of neoangiogenesis. In the present study we report the VEGF expression and its relationship with MVD, measured by CD34, in two lineages of non-small cell lung cancer (NSCL): low differentiated adenocarcinomas and epidermoid carcinomas, in order to consider the possibility of using the correlation between both antibodies as a prognostic factor. Tumor sections were stained by immunohistochemistry for CD34 and VEGF. The results showed that the mean value of VEGF for adenocarcinoma was significantly higher than the one for epidermoid carcinoma (p < 0.001). However, the mean of MVD did not show significant differences between both types of tumors. The conventional factors taken into consideration (age over 60, sex, and presence of lymph nodes) was not significantly related to the angiogenic factors examined. In conclusion, we could affirm that CD34 is a better prognostic marker of neoangiogenesis in NSCLC, because both types of tumors have the same clinical prognosis, and so we expected the same behaviour from both markers.


Impact of housing improvement and the socio-physical environment on the mental health of children's carers: a cohort study in Australian Aboriginal communities.
The mental health of carers is an important proximate factor in the causal web linking housing conditions to child health, as well as being important in its own right. Improved understanding of the nature of the relationships between housing conditions, carer mental health and child health outcomes is therefore important for informing the development of housing programs. This paper examines the relationship between the mental health of the carers of young children, housing conditions, and other key factors in the socio-physical environment. This analysis is part of a broader prospective cohort study of children living in Aboriginal communities in the Northern Territory (NT) of Australia at the time of major new community housing programs. Carer's mental health was assessed using two validated scales: the Affect Balance scale and the Brief Screen for Depression. The quality of housing infrastructure was assessed through detailed surveys. Secondary explanatory variables included a range of socio-environmental factors, including validated measures of stressful life events. Hierarchical regression modelling was used to assess associations between outcome and explanatory variables at baseline, and associations between change in housing conditions and change in outcomes between baseline and follow-up. There was no clear or consistent evidence of a causal relationship between the functional state of household infrastructure and the mental health of carers of young children. The strongest and most consistent associations with carer mental health were the measures of negative life events, with a dose-response relationship, and adjusted odds ratio of over 6 for carers in the highest stress exposure category at baseline, and consistent associations in the follow up analysis. The findings highlight the need for housing programs to be supported by social, behavioral and community-wide environmental programs if potential health gains are to be more fully realized, and for rigorous evaluation of such programs for the purpose of informing future housing initiatives.


Classification of children with ADHD through task-related EEG recordings via Swarm-Decomposition-based Phase Locking Value.
Attention Deficit/Hyperactivity Disorder (ADHD) is a common neurodevelopmental disorder mainly affecting children. ADHD children brain activity is reported to present alterations from neurotypically developed children, yet establishment of an EEG biomarker, which is of high importance in clinical practice and research, has not been achieved. In this work, task-related EEG recordings from 61 ADHD and 60 age-matched non-ADHD children are analyzed to examine the underlying Cross-Frequency Coupling phenomena. The proposed framework introduces personalized brain rhythm extraction in the form of oscillatory modes via Swarm Decomposition, allowing for the transition from sensor-level connectivity to source-level connectivity. Oscillatory modes are then subjected to a phase locking value-based feature extraction and the efficiency of the extracted features in separating ADHD from non-ADHD individuals is evaluated by means of a nested 5-fold cross validation scheme. The experimental results of the proposed framework (Area Under the Receiver Operating Characteristics Curve-AUROC: 0.9166) when benchmarked against the commonly used filter-based brain rhythm extraction (AUROC: 0.8361) underscore its efficiency and demonstrate its overall superiority over other state-of-the-art functional connectivity approaches in this classification task for this dataset.Clinical relevance-This framework provides novel insights about brain regions of interest that are involved in ADHD task-related function and holds promise in providing objective ADHD biomarkers by extending classic sensor-level connectivity to source-level.


Primary liver cancer and occupational exposure.
In a dynamic population-based case-referent study on primary liver cancer and occupational exposure, the work histories of 344 cases and 861 referents, derived from the follow-up of the whole Finnish population in 1976-1978 and 1981, were analyzed by industry, occupation, and agent. After adjustment for alcohol consumption, elevated odds ratios were found for the categories other agricultural workers (mainly milkmaids), clerical workers, persons exposed to welding fumes and those exposed to other inorganic dusts (mainly silica). One possible explanation for the excess among milkmaids was exposure to dust from cattle feed contaminated with aflatoxins. The excess among clerical workers was probably due to nonoccupational factors or chance because occupational exposure was generally rare. The excesses for welding fumes and inorganic dusts, although compatible with occupational etiology, contradict the results of many previous studies carried out among workers exposed to silica dust and welders.


Inhibition of HIV-1 reverse transcriptase and protease by phlorotannins from the brown alga Ecklonia cava.
The bioassay-directed isolation of a marine brown alga, Ecklonia cava, afforded four phlorotannin derivatives, eckol (1), 8,8'-bieckol (2), 8,4"'-dieckol (3), and phlorofucofuroeckol A (4). Among these compounds, 2 and 3 exhibited an inhibitory effect on human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) and protease. Specifically, they inhibited the RT more potently than the protease. The inhibitory activity of compound 2 (IC(50), 0.51 microM) against HIV-1 RT was comparable to that of nevirapine (IC(50), 0.28 microM), a reference compound. An enzyme kinetic assay showed that this compound inhibited the RNA-dependent DNA synthesis activity of HIV-1 RT noncompetitively against dUTP/dTTP with a K(i) value of 0.78 microM. With respect to the homopolymeric template/primer, (rA)n(dT)15, 8,8'-bieckol (2) displayed an uncompetitive type of inhibition (K(i), 0.23 microM).


The long-term use of enalapril and hydrochlorothiazide in two novel mutations patients with Dent's disease type 1.
Dent's disease type 1 is an X-linked tubular disease caused by mutations in the renal chloride channel CLCN-5, and it is characterized by low molecular weight proteinuria, hypercalciuria, nephrocalcinosis, and renal failure. Several cases have been described in which the only presenting symptoms were asymptomatic proteinuria, and focal segmental or global glomerulosclerosis. The renal failure in these patients may be caused by hypercalciuria and persistent proteinuria. Therefore, angiotensin converse enzyme inhibitor and thiazides could be useful. Our aim is to report the effects of these drugs in two novel mutations patients with Dent's disease type 1. In this report, no significant correlations between dosage of hydrochlorothiazide and calciuria and no significant correlations between proteinuria and dosage of enalapril were detected. This is important since these are polyuric patients and these drugs could be dangerous to their renal function.


Surgical margin status after robot assisted radical cystectomy: results from the International Robotic Cystectomy Consortium.
Positive surgical margins at radical cystectomy confer a poor prognosis. We evaluated the incidence and predictors of positive surgical margins in patients who underwent robot assisted radical cystectomy for bladder cancer. Using the International Robotic Cystectomy Consortium database we identified 513 patients who underwent robot assisted radical cystectomy, as done by a total of 22 surgeons at 15 institutions from 2003 to 2009. After stratification by age group, gender, pathological T stage, nodal status, sequential case number and institutional volume logistic regression was used to correlate variables with the likelihood of a positive surgical margin. Of the 513 patients 35 (6.8%) had a positive surgical margin. Increasing 10-year age group, lymph node positivity and higher pathological T stage were significantly associated with an increased likelihood of a positive margin (p = 0.010, <0.001 and p <0.001, respectively). Gender, sequential case number and institutional volume were not significantly associated with margin positivity. The rate of margin positive disease at cystectomy was 1.5% for pT2 or less, 8.8% for pT3 and 39% for pT4 disease. Positive surgical margin rates at robot assisted radical cystectomy for advanced bladder cancer were similar to those in open cystectomy series in a large, multi-institutional, prospective cohort. Sequential case number, a surrogate for the learning curve and institutional volume were not significantly associated with positive margins at robot assisted radical cystectomy.


A comparison of knowledge and attitudes about diet and health among 35- to 75-year-old adults in the United States and Geneva, Switzerland.
This study compared responses of US and Geneva residents to items on analogous questionnaires concerning knowledge and attitudes about diet and health. Comparable data were available from 2 population-based sample surveys: the Cancer Control Supplement of the 1987 National Health Interview Survey and the 1994 Bus Santé 2000 in Geneva, Switzerland. Samples included 10,366 US respondents and 698 Geneva respondents, aged 35 to 74 years. The analysis involved descriptive statistics, contingency tables, and linear regression models. In both the United States and Geneva, health consciousness was greater among women and more highly educated persons than among other groups. Compared with Americans, Genevans assigned more importance to avoiding salt, sugar, and overweight (odds ratio = 1.6, 2.9, and 5.9, respectively) and less importance to lowering cholesterol (odds ratio = 0.6). Genevans were more likely to recognize the relatively high fiber content of lettuce, carrots, and apples. Recognition of low-fat foods was slightly better in the United States. Knowledge and attitudes differed despite high general diet and health awareness in both populations. Identifying why generally similar dietary guidance messages are embraced to different extents across cultures may facilitate global implementation.


Totally asymmetric exclusion process on a ring with internal degrees of freedom.
A totally asymmetric exclusion process on a ring with ν nonconserved internal degrees of freedom, where particles hop forward with a rate that depends on their internal state, has been studied. We show, using a mapping of the model to a zero range process with ν different kinds of boxes, that steady-state weights can be written in a matrix product form and calculate the spatial correlations exactly. A comparison of the model with an equivalent conserved system reveals that unequal hopping rates of particles belonging to different internal states are responsible for the nontrivial correlations.


Modeling larval dispersal for the gilthead seabream in the northwestern Mediterranean Sea.
To investigate dispersal and connectivity between spawning and lagoon nursery habitats of the gilthead seabream, Sparus aurata, in the Gulf of Lions (northwestern Mediterranean Sea), we modeled the potential transport of the species' larvae between its supposed main spawning site in the region (the Planier Island) and two of its main local nursery areas (the coastal lagoons of Thau and Salses-Leucate). Passive larval drift simulations using a dispersal biophysical model showed a large variability in the possible trajectories from spawning to nursery areas and in the predicted ages for larvae arrival on the two nursery sites. The most common ages at arrival obtained in the simulations (20-60 days) are broadly consistent with previous modeling studies but contrast with the actual ages of the S. aurata post-larvae collected in 2016 and 2017 at time of the lagoon entrances (60-90 days, from otolith readings). The period between 25 and 70 days being critical for gilthead seabream larvae to acquire sufficient swimming, osmoregulatory, and olfactory abilities to enter coastal lagoons, we argue that ontogenic development plays a crucial role in the transport and local retention of S. aurata larvae in the studied region, explaining the discrepancy between simulation results and observed data.


Towards flexible organic thin film transistors (OTFTs) for biosensing.
We have studied parylene-N and parylene-C for their use as substrates and gate dielectrics in OTFTs. Parylene-N films with a thickness of 300 nm show the required dielectric properties, as verified by breakthrough-voltage measurements. The surface roughness measured for 300 nm thick parylene-N films is 4-5 nm. However, initial growth of parylene depends on the subjacent surface. This results in different thicknesses on Au electrodes and substrate materials for thin films. Capping of micro-patterned Au-electrodes with a thin Al layer via lift-off results in homogenous parylene film thickness on the whole sample surface. OTFTs are fabricated on glass with parylene-N as a gate dielectric and pentacene as a semiconductor. The electrodes are patterned by photolithography enabling micrometer sized features. The contact resistance is extracted by variation of the channel length. Modification of the parylene dielectric layer surface by plasma treatment with oxygen after deposition allows shifting of the threshold voltage to more positive values, however at the cost of increasing hysteresis. OTFTs fabricated on thin parylene-C films can be peeled off and could result in flexible devices employing parylene-C foil as a substrate. For a foil thickness of 3-4 μm, operational devices can be bent down to radii less than 1 mm, e.g. in the range of cannulas. Operation of such OTFTs with parylene-C as a gate dielectric in liquids is demonstrated. The OTFT current can be modulated by the potential in the electrolyte as well as by the bottom gate potential. This allows for application of such OTFTs as sensors in medical devices.


Silencing RNA-directed RNA polymerase 2 increases the susceptibility of Nicotiana attenuata to UV in the field and in the glasshouse.
RNA-directed RNA-polymerases (RdRs) are essential in small interfering RNA (siRNA) biogenesis and appear to be functionally specialized. We examined the consequences of silencing RdR2 in Nicotiana attenuata with a field release, and transcriptional, two-dimensional proteomic and metabolite analyses. NaRdR2-silenced plants (irRdR2) had large reductions (46% of wild type) in 22-24-nt small RNAs (smRNAs), and smaller reductions (35, 23 and 26% of wild type) in the 19-21, 25-27 and 28-30-nt smRNAs, respectively. When planted into their native habitats in the Great Basin Desert, irRdR2 plants had impaired growth and reproductive output, which were associated with reduced levels of leaf phenolics (rutin and 4'-chlorogenic acid) and MYB and PAL transcripts, but were unaffected in their herbivore resistance. These phenotypes were confirmed in glasshouse experiments, but only when irRdR2 plants were grown with UV-B radiation. irRdR2 plants had wild-type levels of elicited phytohormones and resistance to Manduca sexta attack, but when exposed to UV-B, had reduced growth, fitness, levels of MYB and PAL transcripts, and phenolics. Proteins related to protection against oxidative and physiological stresses, chromatin remodeling and transcription were also downregulated. Silencing the MYB gene by virus-induced gene silencing (VIGS) in wild-type plants reduced levels of PAL transcripts and phenolics, as it did in UV-exposed irRdR2 plants. Bioinformatic analysis revealed that genes involved in phenylpropanoid biosynthesis contained a large number of smRNA binding motives, suggesting that these genes are targets of smRNAs. We conclude that although NaRdR2 transcripts are upregulated in response to both UV-B and herbivore elicitation, the responses they regulate have been tailored to provide protection from UV-B radiation.


Pharmacological and toxicological aspects of 4-demethoxy-3'-deamino-3'-aziridinyl-4'-methylsulphonyl-daunorubicin (PNU-159548): a novel antineoplastic agent.
4-demethoxy-3'-deamino-3'-aziridinyl-4'-methylsulphonyl-daunorubicin (PNU-159548) belongs to a novel class of antitumor compounds (termed alkycyclines) and is currently undergoing Phase II clinical trial. In the present study, we investigated the in vitro and in vivo antitumor activity, the pharmacokinetics, and the toxicological profile of this compound. PNU-159548 showed good cytotoxic activity in murine and human cancer cells growing in vitro, with an average concentration for 50% growth inhibition of 15.8 ng/ml. The drug showed strong antitumor efficacy in vivo after i.v. and p.o. administration against rapidly proliferating murine leukemias and slowly growing transplantable human xenografts. At non-toxic doses, PNU-159548 produced complete regression and cures in ovarian, breast, and human small cell lung carcinomas. Fourteen of 16 models studied, including colon, pancreatic, gastric, and renal carcinomas, astrocytoma and melanoma, were found to be sensitive to PNU-159548. In addition, PNU-159548 was effective against intracranially implanted tumors. Toxicological studies revealed myelosuppression as the main toxicity in both mice and dogs. The maximum tolerated doses, after a single administration, were 2.5 mg/kg of body weight in mice, 1.6 mg/kg in rats, and 0.3 mg/kg in dogs. In the cyclic studies, the maximum tolerated doses were 0.18 mg/kg/day (cumulative dose/cycle: 0.54 mg/kg) in rats and 0.05 mg/kg/day (cumulative dose/cycle: 0.15 mg/kg) in dogs. PNU-159548 showed minimal cardiotoxicity, when compared with doxorubicin in the chronic rat model at a dose level inducing similar myelotoxicity. Animal pharmacokinetics, carried out in mice, rats, and dogs, was characterized by high volumes of distribution, plasma clearance of the same order of the hepatic blood flow, and short terminal half-life. These findings support the conclusion that PNU-159548 is an excellent candidate for clinical trials in the treatment of cancer.


Prospects for improving brain function in individuals with Down syndrome.
Down syndrome (DS), which results from an extra copy of chromosome 21 (trisomy 21), is the most common genetically defined cause of intellectual disability. Although no pharmacotherapy aimed at counteracting the cognitive and adaptive deficits associated with this genetic disorder has been approved at present, there have been several new promising studies on pharmacological agents capable of rescuing learning/memory deficits seen in mouse models of DS. Here, we will review the available mouse models for DS and provide a comprehensive, albeit not exhaustive review of the following preclinical research strategies: (1) SOD1 and antioxidant agents; (2) APP and γ-secretase inhibitors; (3) DYRK1A and the polyphenol epigallocatechin gallate (EGCG); (4) GIRK2 and fluoxetine; (5) adrenergic receptor agonists; (6) modulation of GABAA and GABAB receptors; (7) agonism of the hedgehog signaling pathway; (8) nerve growth factor (NGF) and other neurotrophic factors; (9) anticholinesterase (AChE) agents; and (10) antagonism of NMDA receptors. Finally, we will review briefly five different strategies in DS that have led to clinical studies that either have been concluded or are currently underway: (1) antioxidant therapy; (2) AChE therapy; (3) green tea extract therapy; (4) RG1662 therapy; and (5) memantine therapy. These are exciting times in DS research. Within a decade or so, it is well into the realm of possibility that new forms of pharmacotherapies might become valuable tools in the armamentarium of developmental clinicians, as adjutants to more traditional and proven forms of habilitative interventions aimed at improving the quality of life of individuals with DS.


Cell surface GRP78 promotes stemness in normal and neoplastic cells.
Reliable approaches to identify stem cell mechanisms that mediate aggressive cancer could have great therapeutic value, based on the growing evidence of embryonic signatures in metastatic cancers. However, how to best identify and target stem-like mechanisms aberrantly acquired by cancer cells has been challenging. We harnessed the power of reprogramming to examine GRP78, a chaperone protein generally restricted to the endoplasmic reticulum in normal tissues, but which is expressed on the cell surface of human embryonic stem cells and many cancer types. We have discovered that (1) cell surface GRP78 (sGRP78) is expressed on iPSCs and is important in reprogramming, (2) sGRP78 promotes cellular functions in both pluripotent and breast cancer cells (3) overexpression of GRP78 in breast cancer cells leads to an induction of a CD24-/CD44+ tumor initiating cell (TIC) population (4) sGRP78+ breast cancer cells are enriched for stemness genes and appear to be a subset of TICs (5) sGRP78+ breast cancer cells show an enhanced ability to seed metastatic organ sites in vivo. These collective findings show that GRP78 has important functions in regulating both pluripotency and oncogenesis, and suggest that sGRP78 marks a stem-like population in breast cancer cells that has increased metastatic potential in vivo.


Spatial and temporal assessment of orofacial somatosensory sensitivity: a methodological study.
To evaluate the sensitivity and reproducibility of a multimodal psychophysical technique for the assessment of both spatial and temporal changes in somatosensory function after an infraorbital nerve block. Sixteen healthy volunteers with a mean (+/- SD) age of 22.5 +/- 3.4 years participated in 2 identical experimental sessions separated by 2 weeks. The subjects rated the perceived intensity of standardized nonpainful tactile, painful pinprick, warm, and cold stimuli applied to 25 points in 5 x 5 matrices in the infraorbital region of each side. The reproducibility of single points was tested, and a mean difference of 1.4 +/- 0.5 was found. A 0-50-100 numerical rating scale (NRS) with 50 denoting "just barely painful" was used. A modified ice hockey mask with adjustable settings was developed as a template to allow stimulation of the same points in the 2 sessions. Assessment of somatosensory function was carried out before the injection (baseline) and after 30 and 60 minutes on both the anesthetized and contralateral (control) side. In addition, the applicability of the psychophysical techniques was tested in pilot experiments in 2 patients before maxillary osteotomy and 3 months afterward. The overall analysis of mean NRS scores, number of points, and center-of-gravity coordinates for all stimulus modalities showed no significant main effects of session. Post-hoc tests for all stimulus modalities demonstrated significantly lower mean NRS scores and significantly more points (hyposensitivity) at 30 and 60 minutes postinjection compared to baseline values on the injection side (Tukey tests: P < .002). In the 2 maxillary osteotomy patients, the psychophysical techniques could successfully be applied, and bilateral hyposensitivity to all stimulus modalities was demonstrated at the 3-month follow-up. The present findings indicate that the psychophysical method is sufficiently reproducible, with no major differences between sessions in healthy subjects. All stimulus modalities demonstrated adequate sensitivity. Furthermore, measurement of points in 5 x 5 matrices allowed a spatial description of somatosensory sensitivity. This method may be valuable for studies on changes in somatosensory sensitivity following trauma or orthognathic surgery on the maxilla.


[Tramadol hydrochloride in the treatment of postoperative shivering].
A double blind study of the therapeutic effect of the opioid tramadol hydrochloride (Gruenenthal, Germany) in the treatment of postoperative trembling was undertaken in 2 groups of patients (50 patients in each) versus that of placebo. The results obtained denoted that tramadol at 1-2 mg/kg arrested completely the postoperative trembling or cut significantly its intensity in 49 (98%) patients. Such high efficiency of tramadol as compared to that of other opioids can be explained by its dual mechanism of action. The dependence of an effective tramadol dose on intensity of shivering and on degree of impaired temperature hemostasis was demonstrated. A comparison of hemodynamic parameters observed before and after the administration of tramadol did not reveal any valuable changes in arterial pressure or cardiac beat rate. Mildly intensified sedation was registered in 17 patients, which is typical of all opioids.


'My neighbourhood is fuzzy, not hard and fast': Individual and contextual associations with perceived residential neighbourhood boundaries among ageing Americans.
Neighborhoods are fluid social and spatial constructs that vary by person and place. How do residential neighborhoods shift as people age? This mixed-method study investigates how perceived neighborhood boundaries and size vary by individual and contextual characteristics. Semi-structured interviews with 125 adults aged 55-92 living in the Minneapolis (Minnesota) metropolitan area suggested that neighborhood boundaries are "fuzzy". Qualitative thematic analysis identified duration of residence and housing stability, race, life-space mobility, social capital, sense of safety, and the built and social environment as key neighborhood determinants. This informed quantitative analyses among 7,811 respondents (mean age 72) from the REasons for Geographic And Racial Differences in Stroke (REGARDS) Study who self-reported how many blocks composed their neighborhoods. We tested individual and contextual factors identified in the qualitative results as related to perceived neighborhood size. Three-level gamma regression models showed that being older, white, less educated, lower income, less physically and cognitively healthy, less active, less socially supported, and feeling unsafe were significantly associated with smaller self-reported neighborhood sizes. Further, living in less racially diverse, less dense, and less affluent areas were significantly associated with smaller neighborhoods. The mixed-methods findings deepen understanding of scale in neighborhood-based research, inform urban planning interventions, and help understand what "neighborhood" means among diverse aging Americans.


IQN path ASBL report from the first European cfDNA consensus meeting: expert opinion on the minimal requirements for clinical ctDNA testing.
Liquid biopsy testing is a new laboratory-based method that detects tumour mutations in circulating free DNA (cfDNA) derived from minimally invasive blood sampling techniques. Recognising the significance for clinical testing, in 2017, IQN Path provided external quality assessment for liquid biopsy testing. Representatives of those participating laboratories were invited to attend a workshop to discuss the findings and how to achieve quality implementation of cfDNA testing in the clinical setting, the discussion and outcomes of this consensus meeting are described below. Predictive molecular profiling using tumour tissue in order to select cancer patients eligible for targeted therapy is now routine in diagnostic pathology. If insufficient tumour tissue material is available, in some circumstances, recent European Medicines Agency (EMA) guidance recommends mutation testing with plasma cfDNA. Clinical applications of cfDNA include treatment selection based on clinically relevant mutations derived from pre-treatment samples and the detection of resistant mutations upon progression of the disease. In order to identify tumour-related mutations in amongst other nucleic acid material found in plasma samples, highly sensitive laboratory methods are needed. In the workshop, we discussed the variable approaches taken with regard to cfDNA extraction methods, the tests, and considered the impact of false-negative test results. We explored the lack of standardisation of complex testing procedures ranging from plasma collection, transport, processing and storage, cfDNA extraction, and mutation analysis, to interpretation and reporting of results. We will also address the current status of clinical validation and clinical utility, and its use in current diagnosis. This workshop revealed a need for guidelines on with standardised procedures for clinical cfDNA testing and reporting, and a requirement for cfDNA-based external quality assessment programs.


Contributions of polyclonal malaria, gametocytemia, and pneumonia to infant severe anemia incidence in malaria hyperendemic Pemba, Tanzania.
The causative factors for severe anemia incidence in sub-Saharan Africa are multifactorial. In an observational, longitudinal study of two cohorts of about 300 infants followed-up for six months in a malaria hyperendemic area, the risk factors for severe anemia incidence were clinical malaria and pneumonia, which outweighed nutritional and sociodemographic factors. Severe anemia incidence was 1-2/year at age 2 months, peaked around 6-7/year at age 7-12 months, and decreased back to 1-2/year at age 16-22 months. The age-dependent increase of severe anemia incidence was shown to be parallel to the age-dependent increase of clinical malaria. Previous clinical malaria episodes increased the severe anemia risk by 80%, and gametocyte carriage and pneumonia at prior visit was associated with a six-fold increase and a > 10-fold increase, respectively. The role of pneumonia and malaria as risk factors, and areas for interventions for severe anemia, should not be underestimated.


The role of interstitial collagens in cleft formation of mouse embryonic submandibular gland during initial branching.
An interstitial collagenase was purified from the explant medium of bovine dental pulp and was shown to degrade collagens I and III but not IV and V. The enzyme halted cleft initiation in the epithelium of 12-day mouse embryonic submandibular glands in vitro, indicating the active involvement of interstitial collagens in the branching morphogenesis. Transmission electron microscopic observation of the intact 12-day gland without any clefts showed the scattered localization of a few collagen fibrils at the epithelial-mesenchymal interface of the bulb and also revealed the presence of numerous microfibrils around the stalk. Collagen bundles were regularly seen close to the wavy basal lamina at the bottom of clefts of the intact 13-day gland and 12-day gland cultured for 17 h under normal conditions. Mesenchymal cells were found in the clefts together with the frequent localization of peripheral nerve fibres and capillary endothelial cells. The collagen bundles were more often observed in the 12-day gland cultured in the presence of bovine dental pulp collagenase inhibitor, which had been shown to enhance cleft formation. In contrast, collagen fibrils were rarely found at the epithelial-mesenchymal interface of the 12-day gland cultured in the presence of Clostridial or bovine dental pulp collagenase. The findings indicated that the formation of interstitial collagen bundles is essential to form clefts in the epithelium both in vivo and in vitro.


Microemulsion-assisted preparation of a mesoporous ferrihydrite/SiO2 composite for the efficient removal of formaldehyde from air.
Mesoporous ferrihydrite/SiO2 composites were synthesized according to a water-in-oil microemulsion method and characterized by X-ray diffraction, scanning electron microscopy, transmission electron microscopy, Fourier transform infrared spectroscopy, thermogravimetry, nitrogen-adsorption/desorption, and by X-ray photoelectron spectroscopy. The as-prepared porous ferrihydrite/SiO2 composites showed an excellent adsorption performance for formaldehyde (HCHO) removal from indoor air at ambient temperature. It was found that the aging time during the synthesis had a significant impact on the pore structure, surface area, and HCHO adsorption of these materials. The ferrihydrite/SiO2 composite that was aged for 3 h in the presence of tetraethyl orthosilicate (TEOS) exhibited a relatively high HCHO adsorption capacity, as well as good recyclability, which was attributed to a relatively large BET surface area, optimal pore size, a suitable Si/Fe atomic ratio, and a synergistic effect between ferrihydrite and SiO2. This work not only demonstrates that porous ferrihydrite/SiO2 composites can act as an efficient adsorbent toward HCHO, but suggests a new route for the rational design of cost-effective and environmentally benign adsorbents with high performance for indoor air purification.


Follow that fish: Uncovering the hidden blue economy in coral reef fisheries.
Despite their importance for human well-being, nearshore fisheries are often data poor, undervalued, and underappreciated in policy and development programs. We assess the value chain for nearshore Hawaiian coral reef fisheries, mapping post-catch distribution and disposition, and quantifying associated monetary, food security, and cultural values. We estimate that the total annual value of the nearshore fishery in Hawai'i is $10.3-$16.4 million, composed of non-commercial ($7.2-$12.9 million) and commercial ($2.97 million licensed + $148,500-$445,500 unlicensed) catch. Hawaii's nearshore fisheries provide >7 million meals annually, with most (>5 million) from the non-commercial sector. Over a third (36%) of meals were planktivores, 26% piscivores, 21% primary consumers, and 18% secondary consumers. Only 62% of licensed commercial catch is accounted for in purchase reports, leaving 38% of landings unreported in sales. Value chains are complex, with major buyers for the commercial fishery including grocery stores (66%), retailers (19%), wholesalers (14%), and restaurants (<1%), who also trade and sell amongst themselves. The bulk of total nearshore catch (72-74%) follows a short value chain, with non-commercial fishers keeping catch for household consumption or community sharing. A small amount (~37,000kg) of reef fish-the equivalent of 1.8% of local catch-is imported annually into Hawai'i, 23,000kg of which arrives as passenger luggage on commercial flights from Micronesia. Evidence of exports to the US mainland exists, but is unquantifiable given existing data. Hawaiian nearshore fisheries support fundamental cultural values including subsistence, activity, traditional knowledge, and social cohesion. These small-scale coral reef fisheries provide large-scale benefits to the economy, food security, and cultural practices of Hawai'i, underscoring the need for sustainable management. This research highlights the value of information on the value chain for small-scale production systems, making the hidden economy of these fisheries visible and illuminating a range of conservation interventions applicable to Hawai'i and beyond.


Spectral Properties of Brain Activity Under Two Anesthetics and Their Potential for Inducing Natural Sleep in Birds.
Both mammals and birds exhibit two sleep states, slow wave sleep (SWS) and rapid eye movement (REM) sleep. Studying certain aspects of sleep-related electrophysiology in freely behaving animals can present numerous methodological constraints, particularly when even fine body movements interfere with electrophysiological signals. Interestingly, under light general anesthesia, mammals and birds also exhibit slow waves similar to those observed during natural SWS. For these reasons, slow waves occurring under general anesthesia are commonly used in the investigation of sleep-related neurophysiology. However, how spectral properties of slow waves induced by anesthesia correspond to those occurring during natural SWS in birds has yet to be investigated systematically. In this study, we systematically analyzed spectral properties of electroencephalographic (EEG) patterns of pigeons (Columba livia) occurring under two commonly used anesthetics, isoflurane and urethane. These data were compared with EEG patterns during natural sleep. Slow waves occurring during spontaneous SWS, and those induced with isoflurane and urethane all showed greatest absolute power in the slowest frequencies (<3 Hz). Isoflurane and urethane-induced slow waves had near-identical power spectra, and both had higher mean power than that observed during SWS for all frequencies examined (0-25 Hz). Interestingly, burst suppression EEG activity observed under deeper planes of isoflurane anesthesia could occur bihemispherically or unihemispherically. Electrophysiological patterns while under isoflurane and urethane share phenomenological and spectral similarities to those occurring during SWS, notably the generation of high amplitude, slow waves, and peak low-frequency power. These results build upon other studies which suggest that some anesthetics exert their effects by acting on natural sleep pathways. As such, anesthesia-induced slow waves appear to provide an acceptable model for researchers interested in investigating sleep-related slow waves utilizing electrophysiological methods not suitable for use in freely behaving birds.


Multiscale mapping of species diversity under changed land use using imaging spectroscopy.
Land use changes are one of the most important factors causing environmental transformations and species diversity alterations. The aim of the current study was to develop a geoinformatics-based framework to quantify alpha and beta diversity indices in two sites in Israel with different land uses, i.e., an agricultural system of fruit orchards, an afforestation system of planted groves, and an unmanaged system of groves. The framework comprises four scaling steps: (1) classification of a tree species distribution (SD) map using imaging spectroscopy (IS) at a pixel size of 1 m; (2) estimation of local species richness by calculating the alpha diversity index for 30-m grid cells; (3) calculation of beta diversity for different land use categories and sub-categories at different sizes; and (4) calculation of the beta diversity difference between the two sites. The SD was classified based on a hyperspectral image with 448 bands within the 380-2500 nm spectral range and a spatial resolution of 1 m. Twenty-three tree species were classified with high overall accuracy values of 82.57% and 86.93% for the two sites. Significantly high values of the alpha index characterize the unmanaged land use, and the lowest values were calculated for the agricultural land use. In addition, high values of alpha indices were found at the borders between the polygons related to the "edge-effect" phenomenon, whereas low alpha indices were found in areas with high invasion species rates. The beta index value, calculated for 58 polygons, was significantly lower in the agricultural land use. The suggested framework of this study succeeded in quantifying land use effects on tree species distribution, evenness, and richness. IS and spatial statistics techniques offer an opportunity to study woody plant species variation with a multiscale approach that is useful for managing land use, especially under increasing environmental changes.


Clinical pathology reference intervals for an in-water population of juvenile loggerhead sea turtles (Caretta caretta) in Core Sound, North Carolina, USA.
The loggerhead sea turtle (Caretta caretta) is found throughout the waters of the Atlantic, Pacific, and Indian Oceans. It is a protected species throughout much of its range due to threats such as habitat loss, fisheries interactions, hatchling predation, and marine debris. Loggerheads that occur in the southeastern U.S. are listed as "threatened" on the U.S. Endangered Species List, and receive state and federal protection. As part of an on-going population assessment conducted by the National Marine Fisheries Service, samples were collected from juvenile loggerhead sea turtles in Core Sound, North Carolina, between 2004 and 2007 to gain insight on the baseline health of the threatened Northwest Atlantic Ocean population. The aims of the current study were to establish hematologic and biochemical reference intervals for this population, and to assess variation of the hematologic and plasma biochemical analytes by season, water temperature, and sex and size of the turtles. Reference intervals for the clinical pathology parameters were estimated following Clinical Laboratory Standards Institute guidelines. Season, water temperature, sex, and size of the turtles were found to be significant factors of variation for parameter values. Seasonal variation could be attributed to physiological effects of decreasing photoperiod, cooler water temperature, and migration during the fall months. Packed cell volume, total protein, and albumin increased with increasing size of the turtles. The size-related differences in analytes documented in the present study are consistent with other reports of variation in clinical pathology parameters by size and age in sea turtles. As a component of a health assessment of juvenile loggerhead sea turtles in North Carolina, this study will serve as a baseline aiding in evaluation of trends for this population and as a diagnostic tool for assessing the health and prognosis for loggerhead sea turtles undergoing rehabilitation.


[An efficient high-throughput screening assay against nuclear transport].
Nuclear transport, an essential cellular process of eukaryotic cells, plays critical roles in differentiation , development, as well as viral disease and oncogenesis, making the small molecules that target this process very intriguing not only in fundamental research but also in disease treatment. However, only one compound, Leptomycin B, is commercially available to inhibit nuclear transport. Therefore, it will be a great advantage to establish an assay that targets the whole nuclear transport pathway for screening and obtaining small molecule that regulates nuclear transport. In this study, we established an efficient nuclear transport assay based on the reconstitution of GFP by NZGFP and CZGFP. We constructed NZGFP-NES and CZGFP-NLS, making them locate in cytoplasm and nucleus separately. Their distribution will be changed when nuclear transport is interfered, resulting in co-localization of NZGFP-NES and CZGFP-NLS and subsequent reconstitution of fluorescent GFP. The inhibiting effect of Leptomycin B on nuclear transport can be sensitively detected by NZGFP-NES/CZGFP-NLS report system, providing an efficient assay for high-throughput screening of small molecule against nuclear transport.


Relationships between sleep quality and brain volume, metabolism, and amyloid deposition in late adulthood.
Recent studies in mouse models of Alzheimer's disease (AD) and in humans suggest that sleep disruption and amyloid-beta (Aβ) accumulation are interrelated, and may, thus, exacerbate each other. We investigated the association between self-reported sleep variables and neuroimaging data in 51 healthy older adults. Participants completed a questionnaire assessing sleep quality and quantity and underwent positron emission tomography scans using [18F]florbetapir and [18F]fluorodeoxyglucose and an magnetic resonance imaging scan to measure Aβ burden, hypometabolism, and atrophy, respectively. Longer sleep latency was associated with greater Aβ burden in prefrontal areas. Moreover, the number of nocturnal awakenings was negatively correlated with gray matter volume in the insular region. In asymptomatic middle-aged and older adults, lower self-reported sleep quality was associated with greater Aβ burden and lower volume in brain areas relevant in aging and AD, but not with glucose metabolism. These results highlight the potential relevance of preserving sleep quality in older adults and suggest that sleep may be a factor to screen for in individuals at risk for AD.


Surgical manipulation of mammalian embryos in vitro.
Whole-embryo culture systems are useful in the fields of not only embryology but also teratology, toxicology, pharmacology, and physiology. Of the many advantages of whole-embryo culture, we focus here on the surgical manipulation of mammalian embryos. Whole-embryo culture allows us to manipulate mammalian embryos, similarly to fish, amphibian and avian embryos. Many surgical experiments have been performed in mammalian embryos in vitro. Such surgical manipulation alters the destiny of morphogenesis of the embryos and can answer many questions concerning developmental issues. As an example of surgical manipulation using whole-embryo culture systems, one of our experiments is described. Microsurgical electrocauterization of the deep preaxial mesodermal programmed cell death zone (fpp) in the footplate prevented the manifestation of polydactyly in genetic polydactyly mouse embryos (Pdn/Pdn), in which fpp was abolished.


Synthesis of mixed phosphorotrithioates via thiol coupling with bis(diisopropylamino)chlorophosphine and sulphenyl chloride.
In this study, we report a novel and efficient one-pot synthesis of mixed phosphorotrithioates under mild conditions at ambient temperature, obviating the requirement for supplementary additives. The method's versatility stems from its utilization of diverse thiols as nucleophilic reactants, 1-chloro-N,N,N',N'-tetraisopropylphosphanediamine [bis(diisopropylamino)chlorophosphine] as the phosphorus precursor, and various sulphenyl chlorides as sources of electrophilic sulfur. Notably, our investigation extends beyond mixed phosphorotrithioates to encompass the synthesis of phosphoroselenodithioates, underscoring the broad applicability of this synthetic protocol.


Physical activity-key issues in treatment of childhood obesity.
Changes in physical activity with the aim of increasing energy expenditure are usually an important component of childhood obesity treatment. Physical activity also has several other aspects that are positive for the obese child's health, such as improving the metabolic profile and psychological well being. The aim of this paper is to give a short review of what we know about physical activity in paediatric obesity treatment. In addition, practical recommendations will be presented which a health care provider can suggest to obese children and their families with a special focus on daily activity, participation in physical education classes and sports, sedentary behaviours, active commuting to school and how to get family and friends involved in supporting the child.


SingleScan: a comprehensive resource for single-cell sequencing data processing and mining.
Single-cell sequencing has shed light on previously inaccessible biological questions from different fields of research, including organism development, immune function, and disease progression. The number of single-cell-based studies increased dramatically over the past decade. Several new methods and tools have been continuously developed, making it extremely tricky to navigate this research landscape and develop an up-to-date workflow to analyze single-cell sequencing data, particularly for researchers seeking to enter this field without computational experience. Moreover, choosing appropriate tools and optimal parameters to meet the demands of researchers represents a major challenge in processing single-cell sequencing data. However, a specific resource for easy access to detailed information on single-cell sequencing methods and data processing pipelines is still lacking. In the present study, an online resource called SingleScan was developed to curate all up-to-date single-cell transcriptome/genome analyzing tools and pipelines. All the available tools were categorized according to their main tasks, and several typical workflows for single-cell data analysis were summarized. In addition, spatial transcriptomics, which is a breakthrough molecular analysis method that enables researchers to measure all gene activity in tissue samples and map the site of activity, was included along with a portion of single-cell and spatial analysis solutions. For each processing step, the available tools and specific parameters used in published articles are provided and how these parameters affect the results is shown in the resource. All information used in the resource was manually extracted from related literature. An interactive website was designed for data retrieval, visualization, and download. By analyzing the included tools and literature, users can gain insights into the trends of single-cell studies and easily grasp the specific usage of a specific tool. SingleScan will facilitate the analysis of single-cell sequencing data and promote the development of new tools to meet the growing and diverse needs of the research community. The SingleScan database is publicly accessible via the website at http://cailab.labshare.cn/SingleScan .


Investigating Recovery After Subarachnoid Hemorrhage With the Imaging, Cognition and Outcome of Neuropsychological Functioning After Subarachnoid Hemorrhage (ICONS) Study: Protocol for a Longitudinal, Prospective Cohort Study.
A subarachnoid hemorrhage is a hemorrhage in the subarachnoid space that is often caused by the rupture of an aneurysm. Patients who survive a subarachnoid hemorrhage have a high risk of complications and a negative long-term outcome. The aim of the Imaging, Cognition and Outcome of Neuropsychological functioning after Subarachnoid hemorrhage (ICONS) study is to investigate whether and to what extent deficits exist in multiple domains after subarachnoid hemorrhage, including cognition, emotion and behavior, and to investigate whether brain damage can be detected in patients with subarachnoid hemorrhage. We aim to determine which early measures of cognition, emotion and behavior, and brain damage in the subacute stage play a role in long-term recovery after subarachnoid hemorrhage. Recovery is defined as functioning at a societal participation level, with a focus on resuming and maintaining work, leisure activities, and social relationships over the long term. The ICONS study is an observational, prospective, single-center cohort study. The study includes patients with subarachnoid hemorrhage admitted to the Neurosurgery Unit of the University Medical Centre Groningen in the Netherlands. The inclusion criteria include diagnosis of an aneurysmal subarachnoid hemorrhage or an angiographically negative subarachnoid hemorrhage, sufficient ability in the Dutch language, and age older than 18 years. Patients will undergo neuropsychological assessment and magnetic resonance imaging 6 months after the subarachnoid hemorrhage. Furthermore, patients will be asked to fill in questionnaires on multiple psychosocial measures and undergo a structured interview at 6 months, 1 year, and 2 years after the subarachnoid hemorrhage. The primary outcome measure of the ICONS study is societal participation 1 year after the subarachnoid hemorrhage, measured with the Dutch version of the Impact on Participation and Autonomy questionnaire. The study was launched in December 2019 and recruitment is expected to continue until June 2023. At the time of the acceptance of this paper, 76 patients and 69 healthy controls have been included. The first results are expected in early 2023. The ICONS study is the first to collect and combine data after subarachnoid hemorrhage in a variety of domains, including cognition, emotion and behavior, and brain damage. The results will contribute to a more comprehensive understanding of the consequences of both aneurysmal subarachnoid hemorrhage and angiographically negative subarachnoid hemorrhage, which may ultimately optimize timely treatment for this patient group by setting realistic and attainable goals to improve daily functioning. Netherlands Trial Register NL7803; https://trialsearch.who.int/Trial2.aspx?TrialID=NL7803. DERR1-10.2196/38190.


Evolution of the thyroid hormone-binding protein, transthyretin.
Transthyretin (TTR) belongs to a group of proteins, which includes thyroxine-binding globulin and albumin, that bind to and transport thyroid hormones in the blood. TTR is also indirectly implicated in the carriage of vitamin A through the mediation of retinol-binding protein (RBP). It was first identified in 1942 in human serum and cerebrospinal fluid and was formerly called prealbumin for its ability to migrate faster than serum albumin on electrophoresis of whole plasma. It is a single polypeptide chain of 127 amino acids (14,000 Da) and is present in the plasma as a tetramer of noncovalently bound monomers. The major sites of synthesis of TTR in eutherian mammals, marsupials, and birds are the liver and choroid plexus but in reptiles it is synthesised only in the choroid plexus. The observation that TTR is strongly expressed in the choroid plexus but not in the liver of the stumpy-tailed lizard and the strong conservation of expression in the choroid plexus from reptiles to mammals have been taken as evidence to suggest that extrahepatic synthesis of TTR evolved first. The identification and cloning of TTR from the liver of an amphibian, Rana catesbeiana, and a teleost fish, Sparus aurata, and its absence from the choroid plexus of both species suggest an alternative model for its evolution. Protein modelling studies are presented that demonstrate differences in the electrostatic characteristics of the molecule in human, rat, chicken, and fish, which may explain why, in contrast to TTR from human and rat, TTR from fish and birds preferentially binds triiodo-l-thyronine.


Brassica cover crops for nitrogen retention in the Mid-Atlantic coastal plain.
Brassica cover crops are new to the mid-Atlantic region, and limited information is available on their N uptake capabilities for effective N conservation. Forage radish (Raphanus sativus L. cv. Daikon), oilseed radish (Raphanus sativus L. cv. Adagio), and rape (Brassica napus L. cv. Dwarf Essex) were compared with rye (Secale cereale L. cv. Wheeler), a popular cover crop in the region, with regard to N uptake ability and potential to decrease N leaching at two sites in Maryland. Plants were harvested in fall and spring for dry matter and N analysis. Soil samples from 0 cm to 105 to 180 cm depth were obtained in fall and spring for NH(4)-N and NO(3)-N analyses. Ceramic cup tension lysimeters were installed at depths of 75 to 120 cm to monitor NO(3)-N in soil pore water. Averaged across 3 site-years, forage radish and rape shoots had greater dry matter production and captured more N in fall than rye shoots. Compared with a weedy fallow control, rape and rye caused similar decreases in soil NO(3)-N in fall and spring throughout the sampled profile. Cover crops had no effect on soil NH(4)-N. During the spring on coarse textured soil, pore water NO(3)-N concentrations in freeze-killed Brassica (radish) plots were greater than in control and overwintering Brassica (rape) and rye plots. On fine textured soil, all cover crops provided a similar decrease in pore water NO(3)-N concentration compared with control. On coarse textured soils, freeze-killed Brassica cover crops should be followed by an early-planted spring main crop.


[Reutilization of disposable syringes: frequency and costs for the administration of insulin at home].
The present survey has the purpose to analyze the frequency and costs of the re-utilization of disposable syringes for insulin administration at home. One hundred and thirteen patients with diabetes assisted by a public hospital in a city in the interior São Paulo State, Brazil, were interviewed. The findings showed that regarding the studied population, the frequency of needles re-utilization varied from 1 to 20 times, and syringes re-utilization from 1 to 60 times. The greatest frequencies in the use of the same syringe or needle were of four times followed by three times. Re-utilizing the same syringe four times resulted in a cost reduction of 74.68%. The results pointed out the need for a legislation review as well as for new studies on the risks and benefits arising from this practice.


Generation of rat pancreas in mouse by interspecific blastocyst injection of pluripotent stem cells.
The complexity of organogenesis hinders in vitro generation of organs derived from a patient's pluripotent stem cells (PSCs), an ultimate goal of regenerative medicine. Mouse wild-type PSCs injected into Pdx1(-/-) (pancreatogenesis-disabled) mouse blastocysts developmentally compensated vacancy of the pancreatic "developmental niche," generating almost entirely PSC-derived pancreas. To examine the potential for xenogenic approaches in blastocyst complementation, we injected mouse or rat PSCs into rat or mouse blastocysts, respectively, generating interspecific chimeras and thus confirming that PSCs can contribute to xenogenic development between mouse and rat. The development of these mouse/rat chimeras was primarily influenced by host blastocyst and/or foster mother, evident by body size and species-specific organogenesis. We further injected rat wild-type PSCs into Pdx1(-/-) mouse blastocysts, generating normally functioning rat pancreas in Pdx1(-/-) mice. These data constitute proof of principle for interspecific blastocyst complementation and for generation in vivo of organs derived from donor PSCs using a xenogenic environment.


Exertional desaturation in patients with chronic obstructive pulmonary disease.
Although the Centers for Medicare and Medicaid Services oxygen prescription guidelines utilize a threshold arterial oxygen tension <or=55 mmHg or an oxygen saturation <or=88%, a range of oxygen levels and relative declines have been used in investigations of exertional desaturation in patients with chronic obstructive pulmonary disease (COPD). There is no uniform definition of exertional hypoxemia or standardized exercise protocol to elicit decreases in oxygen levels in individuals with COPD. The causes for exertional desaturation in patients with COPD are multifactorial with ventilation-perfusion mismatching, diffusion-type limitation, shunting and reduced oxygen content of mixed venous blood all contributing to some degree. Neither resting oxygen saturation nor pulmonary function studies can reliably predict which patients with COPD will develop exertional desaturation. However, preserved pulmonary function, especially diffusing capacity, reliably predicts which patients with COPD will sustain oxygenation during exercise. Although exertional desaturation in patients with COPD appears to portend a poor prognosis, there is no evidence that maintenance of normoxemia during exercise improves the survival of these patients. Studies of the effect of supplemental oxygen on exercise performance in individuals with COPD who desaturate with exertion have yielded conflicting results. The use of short-term or "burst" oxygen either prior to or after exertion may not have significant clinical benefit. Differences in the definition of desaturation, mode of exercise, and characteristics of the patient population make it difficult to compare studies of exertional desaturation and its treatment and to determine their applicability to clinical practice.


[Plasma lipoprotein absorption by steroid-producing organs in the rat].
A study was made of the blood plasma iodinated lipoprotein (LP) absorption by the organs and tissues of rats following i.v. injection. The adrenals and ovaries were shown to effectively absorb high density lipoproteins (HDL) as compared to the liver, spleen, kidneys, heart and lungs. HDL absorption reached its maximum 30 min after LP injection. Six hours later the adrenal tissue radioactivity level decreased by 30% and after 12 h 3-fold. The time-course of HDL absorption by the ovaries was characterized by a slow increase in radioactivity after 6 h and a gradual decrease of the label index after 12 h. Differences in HDL absorption by the adrenals and ovaries reflected some functional features of these steroid producing organs.


In vivo binding and hierarchy of assembly of the yeast RNA polymerase I transcription factors.
Transcription by RNA polymerase I in Saccharomyces cerevisiae requires a series of transcription factors that have been genetically and biochemically identified. In particular, the core factor (CF) and the upstream activation factor (UAF) have been shown in vitro to bind the core element and the upstream promoter element, respectively. We have analyzed in vivo the DNAse I footprinting of the 35S promoter in wild-type and mutant strains lacking one specific transcription factor at the time. In this way we were able to unambiguously attribute the protections by the CF and the UAF to their respective putative binding sites. In addition, we have found that in vivo a binding hierarchy exists, the UAF being necessary for CF binding. Because the CF footprinting is lost in mutants lacking a functional RNA polymerase I, we also conclude that the final step of preinitiation-complex assembly affects binding of the CF, stabilizing its contact with DNA. Thus, in vivo, the CF is recruited to the core element by the UAF and stabilized on DNA by the presence of a functional RNA polymerase I.


Green analytical method for the simultaneous analysis of cytochrome P450 probe substrates by poly(N-isopropylacrylamide)-based temperature-responsive chromatography.
High-performance liquid chromatography (HPLC) is the most common analytical method practiced in various fields and used for analysis of almost all drug compounds in the pharmaceutical industries. During drug development, an evaluation of potential drug interaction with cytochrome P450 (CYP) is essential. A "cocktail" approach is often used in drug development to evaluate the effect of a drug candidate on multiple CYP enzymes in a single experiment. So far, simultaneous analysis of multiple CYP substrates, which have greatly different structure and physicochemical properties, has required organic solvents and mobile phase gradient methods. However, despite the recent emphasis on environmental protection, analytical methods that use only aqueous solvents without the use of organic solvents for separation have not been studied well. This study sought to develop the simultaneous analysis of multiple CYP substrates by using poly(N-isopropylacrylamide) (PNIPAAm)-based temperature-responsive chromatography with only aqueous solvents and isocratic methods. Good separation of multiple CYP substrates was achieved without using organic solvents and any gradient methods by temperature-responsive chromatography utilizing a P(NIPAAm-co-n-butyl methacrylate (BMA))- and P(NIPAAm-co-N-acryloyl L-tryptophan methyl ester (L-Trp-OMe))-grafted silica column. Overall, PNIPAAm-based temperature-responsive chromatography represents a remarkably simple, versatile, and environmentally friendly bioanalytical method for CYP substrates and their metabolites.


Heterogeneous spectrum of coreceptor usage among variants within a dualtropic human immunodeficiency virus type 1 primary-isolate quasispecies.
Human immunodeficiency virus type 1 (HIV-1) variants that use the coreceptor CCR5 for entry (R5; macrophage tropic) predominate in early infection, while variants that use CXCR4 emerge during disease progression. Some late-stage variants use CXCR4 alone (X4; T-cell tropic), while others use both CXCR4 and CCR5 (R5X4; dualtropic). It has been proposed that dualtropic R5X4 strains are intermediates in the evolution from R5 to X4, and we hypothesized that a dualtropic primary-isolate quasispecies might contain variants that represent the spectrum of coreceptor use in vivo. We generated a panel of 35 functional full-length env clones from the primary-isolate quasispecies of a dualtropic prototype strain, HIV-1 89.6(PI). Thirty of the functional env clones (86%) were R5X4, four (11%) were R5, and one (3%) was predominantly X4. V3 to V5 sequences did not reveal clustering by coreceptor usage, and no specific sequence motif or V3 charge pattern corresponded to coreceptor utilization. Complete sequencing of seven functionally divergent Env proteins revealed > or =98.7% homology and conservation of structurally important domains. Chimeras between the R5X4 89.6 prototype and an R5 variant indicated that multiple regions contributed to the use of CXCR4, while chimeras with the X4 variant implicated a single residue in V4 in CCR5 use. These results confirm, at the molecular level, both that dualtropic variants are a predominant component of late-stage syncytium-inducing isolates and that variants restricted to each coreceptor coexist with dualtropic species in vivo. Coreceptor-restricted minority variants may reflect residual R5 species from earlier in disease as well as emerging X4 variants.


The repressor which binds the -75 GATA motif of the GPB promoter contains Ku70 as the DNA binding subunit.
Glycophorin B (GPB) is an abundant cell surface glycoprotein which is only expressed in human erythroid cells. Previous functional analysis demonstrated that the repression of the GPB promoter is determined by the binding of a ubiquitous factor which recognizes a GATA motif centered at position -75. In erythroid cells this ubiquitous factor is displaced by the binding of the erythroid-specific factor hGATA1. Here, we have identified the Ku70 protein as a candidate GPB repressor DNA binding subunit through the screening of a human HeLa expression library using the -75 GATA sequence as bait (one-hybrid method). Electrophoretic mobility shift assays demonstrated that the ubiquitous factor that binds the -75 GATA sequence was the Ku70-Ku80 (Ku) heterodimer. Co-transfection experiments demonstrated that overexpression of Ku70 in the K562 erythroleukeamic cell line resulted in transcriptional repression of the chloramphenicol acetyltransferase reporter gene when placed under the control of the wild-type GPB promoter. Conversely, no repression was observed when a mutation that abolished the binding of Ku was introduced in the GPB promoter construct. Altogether, these results indicate that Ku binds in vivo to the -75 WGATAR motif and is involved in negative regulation of the GPB promoter. These findings suggest that, besides its role in many functions, Ku is also involved in transcriptional regulation of erythroid genes.


A prescription for the Epley maneuver: www.youtube.com?
Video-sharing Web sites are being used for information about common conditions including dizziness. The Epley maneuver (EM) is a simple and effective treatment for benign paroxysmal positional vertigo (BPPV) of the posterior canal. However, the maneuver is underused in routine care. In this study, we aimed to describe and analyze the available information about the EM on youtube.com. A YouTube search was performed on August 31, 2011, for videos that demonstrated the entire EM. Detailed data were abstracted from each video and corresponding Web site. Videos were rated on the accuracy of the maneuver by 2 authors, with differences resolved by adjudication. Comments posted by viewers were assessed for themes regarding video use. Of the 3,319 videos identified, 33 demonstrated the EM. The total number of hits for all videos was 2,755,607. The video with the most hits (802,471) was produced by the American Academy of Neurology. Five of the videos accounted for 85% of all the hits. The maneuver demonstration was rated as accurate in 64% (21) of the videos. Themes derived from the 424 posted comments included patients self-treating with the maneuver after reviewing the videos, and providers using the videos as a prescribed treatment or for educational purposes. Accurate video demonstration of the Epley maneuver is available and widely viewed on YouTube. Video-sharing media may be an important way to disseminate effective interventions such as the EM. The impact of video Web sites on outcomes and costs of care is not known and warrants future study.


Radiographic Evaluation of Valvular Heart Disease With Computed Tomography and Magnetic Resonance Correlation.
Valvular heart disease is a group of complex entities with varying etiologies and clinical presentations. There are a number of imaging tools available to supplement clinical evaluation of suspected valvular heart disease, with echocardiography being the most common and clinically established, and more recent emergence of computed tomography and magnetic resonance imaging as additional supportive techniques. Yet even with these newer and more sophisticated modalities, chest radiography remains one of the earliest and most common diagnostic examinations performed during the triage of patients with suspected cardiac dysfunction. Recognizing the anatomic and pathologic features of cardiac radiography including the heart's adaptation to varying hemodynamic changes can provide clues to the radiologist regarding the underlying etiology. In this article, we will elucidate several principles relating to chamber modifications in response to pressure and volume overload as well as radiographic appearances associated with pulmonary fluid status and cardiac dysfunction. We will also present a pattern approach to optimize analysis of the chest radiograph for valvular heart disease, which will help guide the radiologist down a differential diagnostic pathway and create a more meaningful clinical report.


Examination of the Effects of Domestic Water Buffalo (Bubalus bubalis) Grazing on Wetland and Dry Grassland Habitats.
In nature conservation today, there is a global problem with the aggressive expansion of invasive plant species and the conservation of valuable grassland vegetation. Based on this, the following question has been formed: Is the domestic water buffalo (Bubalus bubalis) appropriate for managing various habitat types? How does grazing by water buffalo (Bubalus bubalis) affect on grassland vegetation? This study was carried out in four areas of Hungary. One of the sample areas was in the Mátra Mountains, on dry grassland areas where grazing had been applied for two, four and six years. The other sample areas were in the Zámolyi Basin, where wet fens with a high risk of Solidago gigantea and in a typic Pannonian dry grassland were investigated. In all areas, grazing was carried out with domestic water buffalo (Bubalus bubalis). During the study, we carried out a coenological survey, examining the change of cover of plant species, their feed values and the biomass of the grassland. According to the results, both the number and cover of economically important grasses (from 28% to 34.6%) and legumes (from 3.4% to 25.4%) increased in Mátra as well as the high proportion of shrubs (from 41.8% to 4.4%) shifted toward grassland species. In the areas of the Zámolyi Basin, invasive Solidago has been suppressed completely, the pasture has been converted completely (from 16% to 1%) and the dominant species has become Sesleria uliginosa. Thus, we have found that grazing with buffalo is suitable as a habitat management method in both dry grasslands and wet grasslands. Therefore, in addition to its effectiveness in the control of Solidago gigantea, grazing with buffalo is successful in both nature conservation and economic aspects of grassland vegetation.


Towards new indices of arterial stiffness using systolic pulse contour analysis: a theoretical point of view.
Total arterial stiffness plays a contributory role throughout aging and in numerous cardiovascular diseases, including hypertension. Aortic stiffening is responsible for an increased characteristic impedance (ie, the impedance to the left ventricular pulsatile flow), thus increasing the forward pressure-wave amplitude that contributes to pulse pressure elevation. Aortic stiffening also increases pulse wave velocity, and this results in anticipated and enhanced wave reflections, further augmenting central pulse pressure. Unfortunately, there is no simple time-domain estimate of characteristic impedance. Furthermore, recent guidelines have reviewed the limitations of diastolic pulse contour analysis to estimate arterial stiffness in the time domain. The present theoretical article proposes that systolic pulse contour analysis may provide new, simple time-domain indices quantifying pulsatile load in resting humans. Our proposal was mainly based on 2 simple, validated assumptions: (1) a linear aortic pressure-flow relationship in early systole and (2) a triangular aortic flow wave during systole. This allowed us to describe new time-domain estimates of characteristic impedance, pulsatile load (waveguide ratio), total arterial compliance, and total arterial stiffness. It is demonstrated that total arterial stiffness may be estimated by the following formula: [(Pi - DAP) x ST] / (SV x Deltat), where Pi is the aortic pressure at the inflection point (peak forward pressure wave), DAP is diastolic aortic pressure, ST is systolic ejection time, SV is stroke volume, and Deltat is the time-to-Pi. A mathematical relationship among time intervals and indices of pulsatile load is demonstrated, and the clinical implications are discussed in terms of cardiovascular risk and stroke volume prediction.


Influence of genetic susceptibility on the urinary excretion of 8-hydroxydeoxyguanosine of firefighters.
Oxidative DNA damage has been implicated in carcinogenesis. The DNA damage can be assessed from the urinary excretion of the DNA-repair product 8-hydroxydeoxyguanosine (8-OH-dG). The factors were investigated that influenced the excretion of urinary 8-OH-dG in 78 firefighters. 53 Out of 78 firefighters were exposed to fire within 5 days of the study and 25 were not. 8-OH-dG was measured by ELISA and the distribution of the genotypes of CYP1A1, CYP2E1, GSTM1, and GSTT1 was measured by polymerase chain reaction. The homozygous wild type frequencies of CYP1A1 MspI, CYP1A1 ile-val, CYP2E1, GSTM1, and GSTT1 were 31.5%, 56.2%, 60.3%, 50.7%, and 53.4%, respectively. The geometric mean of urinary 8-OH-dG was 14.1 ng/mg creatinine in more active firefighters and 12.3 ng/mg creatinine in non-exposed and less active subjects. Significantly increased concentrations of urinary 8-OH-dG were found to be associated with cigarette smoking, and 14% of the variation of 8-OH-dG was explained by cigarettes smoked per day. The CYP1A1 MspI, CYP1A1 ile-val, GSTM1, and GSTT1 genetic polymorphisms were not found to be significantly associated with the urinary excretion of 8-OH-dG. However, the subjects carrying the CYP2E1 mutant type excreted higher concentrations of 8-OH-dG and there was a marginally significant interaction of GSTT1 with firefighting activity. Multiple regression analysis confirmed that smoking was the strongest predictor of excretion of 8-OH-dG. Age, body mass index, and firefighting activity were not significant predictive factors for urinary 8-OH-dG. Smoking and CYP2E1 gene polymorphism may be important factors in carcinogenesis and the GSTT1 positive genotype may be a genetic susceptibility factor in firefighters who are exposed regularly to various chemical carcinogens.


Novel immune targets for the treatment of triple-negative breast cancer.
To overcome mechanisms of primary and secondary resistance to the anti-tumor immune response, novel targets such as ICOS, LAG3, and TIM3 are currently being explored at preclinical and early-phase clinical levels. This article examines the landscape of the immune therapeutics investigated in early-phase clinical trials for TNBC. Preclinical rationale is provided for each immune target, predominant expression, and function. Clinical implications and preliminary available trial results are discussed and finally, we reflect on aspects of future expectations and challenges in this field. Several immune strategies have been investigated in TNBC, including co-inhibitory molecules beyond PD1-PD-L1 axis, co-stimulatory checkpoints, cancer vaccines, adoptive cell transfer, combination therapies, as well as different routes of administration. Most of approaches showed signs of anti-cancer activity and a good safety profile in early-phase clinical trials. Since IO provided benefit only to a small subgroup of TNBC patients so far, identifying predictive biomarkers is a priority to refine patient-selection. Data from ongoing clinical trials, with the gradually improving interpretation of the breast tumor immune environment, will hopefully refine the role of new immune targets for the treatment of TNBC.


Gambling in Brazil: a call for an open debate.
To provide an overview of gambling in Brazil, including historical background, past and current legislation, gamblers' profile, treatment and research initiatives. Review of the published literature and research reports. Gambling is deeply rooted in Brazilian history and culture, but gambling regulation historically alternates between legalization and banning. Currently, only state lotteries, horse betting and poker-playing are allowed. There is pressure to widen the games repertoire, but efforts at full legalization and commercial exploration of gambling have receded. Despite the barriers to gambling access, Brazil has prevalence rates similar to other countries: 1.0 and 1.3% life-time prevalence for pathological and problem gambling. A faster progression from regular to problem gambling was found among middle-aged women in clinical samples and for adolescent male gamblers in a population-based sample. Youth gambling is associated with severe forms of gambling and other risk-taking behaviours. Treatment options are scarce, and the public health system is unprepared to support gambling patients. Ongoing incipient efforts are being made to establish a treatment model combining psychotherapy and psychiatric comorbidity treatment with promotion of quality of life. Life-time incidence of pathological gambling in Brazil appears similar to many other countries, at approximately 1%. Arguably, there is a need to develop a more coherent regulatory and treatment response to this societal problem.


Safety, pharmacokinetics, and antitumor activity of the anti-CEACAM5-DM4 antibody-drug conjugate tusamitamab ravtansine (SAR408701) in patients with advanced solid tumors: first-in-human dose-escalation study.
Tusamitamab ravtansine (SAR408701) is an antibody-drug conjugate composed of a humanized monoclonal antibody that binds carcinoembryonic antigen-related cell adhesion molecule-5 (CEACAM5) and a cytotoxic maytansinoid that selectively targets CEACAM5-expressing tumor cells. In this phase I dose-escalation study, we evaluated the safety, pharmacokinetics, and preliminary antitumor activity of tusamitamab ravtansine in patients with solid tumors. Eligible patients were aged ≥18 years, had locally advanced/metastatic solid tumors that expressed or were likely to express CEACAM5, and had an Eastern Cooperative Oncology Group Performance Status of 0 or 1. Patients were treated with ascending doses of tusamitamab ravtansine intravenously every 2 weeks (Q2W). The first three dose levels (5, 10, and 20 mg/m2) were evaluated using an accelerated escalation protocol, after which an adaptive Bayesian procedure was used. The primary endpoint was the incidence of dose-limiting toxicities (DLTs) during the first two cycles, graded using National Cancer Institute Common Terminology Criteria for Adverse Events (NCI CTCAE) v4.03 criteria. Thirty-one patients received tusamitamab ravtansine (range 5-150 mg/m2). The DLT population comprised 28 patients; DLTs (reversible grade 3 microcystic keratopathy) occurred in three of eight patients treated with tusamitamab ravtansine 120 mg/m2 and in two of three patients treated with 150 mg/m2. The maximum tolerated dose was identified as 100 mg/m2. Twenty-two patients (71%) experienced ≥1 treatment-related treatment-emergent adverse event (TEAE), seven patients (22.6%) experienced ≥1 treatment-related grade ≥3 TEAE, and three patients (9.7%) discontinued treatment due to TEAEs. The most common TEAEs were asthenia, decreased appetite, keratopathy, and nausea. Three patients had confirmed partial responses. The mean plasma exposure of tusamitamab ravtansine increased in a dose-proportional manner from 10 to 150 mg/m2. Tusamitamab ravtansine had a favorable safety profile with reversible, dose-related keratopathy as the DLT. Based on the overall safety profile, pharmacokinetic data, and Bayesian model recommendations, the maximum tolerated dose of tusamitamab ravtansine was defined as 100 mg/m2 Q2W.


Interactive effects of global climate change and pollution on marine microbes: the way ahead.
Global climate change has the potential to seriously and adversely affect marine ecosystem functioning. Numerous experimental and modeling studies have demonstrated how predicted ocean acidification and increased ultraviolet radiation (UVR) can affect marine microbes. However, researchers have largely ignored interactions between ocean acidification, increased UVR and anthropogenic pollutants in marine environments. Such interactions can alter chemical speciation and the bioavailability of several organic and inorganic pollutants with potentially deleterious effects, such as modifying microbial-mediated detoxification processes. Microbes mediate major biogeochemical cycles, providing fundamental ecosystems services such as environmental detoxification and recovery. It is, therefore, important that we understand how predicted changes to oceanic pH, UVR, and temperature will affect microbial pollutant detoxification processes in marine ecosystems. The intrinsic characteristics of microbes, such as their short generation time, small size, and functional role in biogeochemical cycles combined with recent advances in molecular techniques (e.g., metagenomics and metatranscriptomics) make microbes excellent models to evaluate the consequences of various climate change scenarios on detoxification processes in marine ecosystems. In this review, we highlight the importance of microbial microcosm experiments, coupled with high-resolution molecular biology techniques, to provide a critical experimental framework to start understanding how climate change, anthropogenic pollution, and microbiological interactions may affect marine ecosystems in the future.


Spectral and temporal detection of blood bilirubin level using a point-of-care device.
Bilirubin is a biomarker for liver inflammation used to assess liver functions. Its concentration in the blood has been measured using a range of techniques both in clinical and point-of-care settings. Existing point-of-care devices utilize a spectral approach, namely dual-wavelength absorption measurement, to assess the blood bilirubin concentration. This work examines a novel temporal approach based on the photodegradation of bilirubin in the blood sample. It demonstrates that combining photodegradation characteristics with dual-wavelength measurement produces a more accurate technique for measuring blood bilirubin concentration. Tracking the evolution of absorbed light as a function of time represents a low-cost and simple way of improving the accuracy of point-of-care devices for bilirubin measurements.Clinical Relevance - This work demonstrates a facile and cheap bilirubin monitoring approach that may allow bilirubin monitoring applications in homes after a patient is discharged from a hospital, which may decrease the burden on patients, families, and clinicians.


Effects of lorcaserin on reinstatement of responding previously maintained by cocaine or remifentanil in rhesus monkeys.
Drug abuse remains a serious public health issue, underscoring the need for additional treatment options. Agonists at serotonin (5-HT)2C receptors, particularly lorcaserin, are being considered as pharmacotherapies for abuse of a variety of drugs, including cocaine and opioids. The current study compared the capacity of lorcaserin to attenuate reinstatement of extinguished responding previously maintained by either cocaine or an opioid; this type of procedure is thought to model relapse, an important aspect of drug abuse. Six rhesus monkeys responded under a fixed-ratio schedule for cocaine (0.032 mg/kg/infusion) or remifentanil (0.00032 mg/kg/infusion). Reinstatement of extinguished responding was examined following administration of noncontingent infusions of cocaine (0.32 mg/kg) or heroin (0.0032-0.1 mg/kg) combined with response-contingent presentations of the drug-associated stimuli, or heroin alone without presentation of drug-associated stimuli. When combined with drug-associated stimuli, cocaine and heroin increased extinguished responding. On average, monkeys emitted fewer reinstated responses following 0.32 mg/kg cocaine, compared with the number of responses emitted when cocaine was available for self-administration or when extinguished responding was reinstated by 0.032 mg/kg heroin. When drug-associated stimuli were not presented, heroin did not increase responding. Lorcaserin dose dependently attenuated reinstated responding, and its potency was similar regardless of whether cocaine or heroin was given before reinstatement sessions. The generality of this effect of lorcaserin across pharmacological classes of abused drugs might make it particularly useful for reducing relapse-related behaviors in polydrug abusers. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


Absolute depth estimation from a single defocused image.
Shape from defocus (SFD) is one of the most popular techniques in monocular 3D vision. While most SFD approaches require two or more images of the same scene captured at a fixed view point, this paper presents an efficient approach to estimate absolute depth from a single defocused image. Instead of directly measuring defocus level of each pixel, we propose to design a sequence of aperture-shape filters to segment a defocused image by defocus level. A boundary-weighted belief propagation algorithm is employed to obtain a smooth depth map. We also give an estimation of depth error. Extensive experiments show that our approach outperforms the state-of-the-art single-image SFD approaches both in precision of the estimated absolute depth and running time.


[Epidural haematoma in a newborn].
Birth injuries occur occasionally but, in contrast, perinatally acquired epidural and intracerebral hematomas as well as neonatal skull fractures are extremely rare. The appropriate diagnostic and therapeutic modalities depend on the size and site of the injury as well as any accompanying diseases. We report the case of a neonatal epidural and intracerebral haematoma with skull fracture following secondary caesarean section. There was no evidence for trauma during pregnancy, birth injury or other possible causes of bleeding. The haematoma was decompressed surgically. One year after surgery the boy suffers from moderate neurological compromise and focal seizures.


Tandem metalloenzymes gate plant cell entry by pathogenic fungi.
Global food security is endangered by fungal phytopathogens causing devastating crop production losses. Many of these pathogens use specialized appressoria cells to puncture plant cuticles. Here, we unveil a pair of alcohol oxidase-peroxidase enzymes to be essential for pathogenicity. Using Colletotrichum orbiculare, we show that the enzyme pair is cosecreted by the fungus early during plant penetration and that single and double mutants have impaired penetration ability. Molecular modeling, biochemical, and biophysical approaches revealed a fine-tuned interplay between these metalloenzymes, which oxidize plant cuticular long-chain alcohols into aldehydes. We show that the enzyme pair is involved in transcriptional regulation of genes necessary for host penetration. The identification of these infection-specific metalloenzymes opens new avenues on the role of wax-derived compounds and the design of oxidase-specific inhibitors for crop protection.


Inhibition kinetics of digestive proteases for Anticarsia gemmatalis.
Anticarsia gemmatalis Hübner, 1818 (Lepidoptera) is a major pest of soybean in the Brazil. It is known that the reduction of proteolytic activity by the ingestion of protease inhibitors reduces digestion and larval development of the insects. Control via inhibition of the digestive enzymes necessitates deeper knowledge of the enzyme kinetics and the characterization of the inhibition kinetics of these proteases, for better understanding of the active centers and action mechanisms of this enzyme. Trypsin-like proteases found in the gut of Anticarsia gemmatalis were purified in a p-aminobenzamidine agarose column. Kinetic characterization showed KM 0.503 mM for the L-BApNA substrate; Vmax= 46.650 nM s-1; Vmax/[E]= 9.256 nM s-1 mg L-1 and Vmax/[E]/KM= 18.402 nM s-1 mg L-1 mM. The Ki values for the inhibitors benzamidine, berenil, SKTI and SBBI were 11.2 µM, 32.4 µM, 0.25 nM and 1.4 nM, respectively, and all revealed linear competitive inhibition. The SKTI showed the greatest inhibition, which makes it a promising subject for future research to manufacture peptide mimetic inhibitors.


Intake energy, energy retention and heat production in lambs from birth to 24 weeks of age.
Intake energy (IE), metabolizable energy intake (MEI), energy retention (ER) and heat production (HP) were estimated in twelve male and six female suckling lambs from birth to 4 wk of age and in nine of these male lambs after weaning from 9 to 24 wk of age. Intake energy and MEI were estimated from the milk intake and combustible energy of the milk in the suckling lambs and from digestibility trials and energy content of feed and feces in the weaned lambs. Energy retention was estimated from body composition changes and HP was calculated from MEI - ER and from the rate of O2 consumption. The O2 consumption of the ewes was also measured during late pregnancy and during lactation. In the suckling lambs, daily MEI was 277 kcal/kg.75 and ER was 112 kcal/kg.75 and as calculated from O2 consumption was 168 kcal/kg.75. Daily HP as calculated by MEI - ER was 165 kcal/kg.75. Daily maintenance HP was equal to 121 kcal/kg.75 and the efficiency of utilization of energy for ER was 70.7%. In the weaned lambs, HP as measured from O2 consumption was 10% lower than that estimated using MEI and ER. Comparison of 9- to 16-wk-old with 17- to 24-wk-old lambs, showed daily increases in MEI from 212 to 228 kcal/kg.75, in ER from 54 to 95 kcal/kg.75 and in HP from 158 to 194 kcal/kg.75, whereas there was a daily decrease in maintenance HP from 101 to 90 kcal/kg.75. Efficiency of utilization of energy for production was similar in the two postweaning periods (48.8 and 46.5%, respectively). The O2 consumption of the ewes (/kg.75) was increased over that of nonpregnant, nonlactating controls by 30% in late pregnancy, by 62% in the first month of lactation and by 50% in the second month of lactation.


Selective alkaline stripping of metal ions after solvent extraction by base-stable 1,2,3-triazolium ionic liquids.
Novel 1,2,3-triazolium ionic liquids with a high base stability were synthesized for use in solvent extraction of first-row transition elements and rare earths from chloride media. The synthesis of these ionic liquids makes use of a recently reported, metal-free multicomponent reaction that allows full substitution of the 1,2,3-triazolium skeleton. The physical and chemical properties of these ionic liquids are compared with those of a trisubstituted analog. Peralkylation of the 1,2,3-triazolium skeleton leads to ionic liquids with superior properties, such as low viscosity, low solubility in water and higher thermal and base stability. Iodide and thiocyanate ionic liquids with peralkylated cations were applied to the solvent extraction of metal ions, and their stability in alkaline media was exploited in the selective stripping of the metals from the loaded ionic liquid phase by alkaline solutions. EXAFS and Raman spectroscopy were performed to gain insight into the extraction mechanism. The applicability of these extraction systems was demonstrated in separations relevant for the recovery of metals from ores and end-of-life products: Fe(iii)/Cu(ii)/Zn(ii) (copper ores, brass scraps) and Fe(iii)/Nd(iii) (rare earth magnets).


Novel PFAS-specific monitoring approach for highly impacted surface waters.
In regulatory environmental monitoring programs, only a very small fraction of the vast number of per- and polyfluoroalkyl substances (PFAS) are investigated by target analysis. Therefore, non-target analysis (NTA) studies are increasingly conducted to detect unknown or unnoticed PFAS. These studies are often based on a few grab samples. Thus, discontinuously emitted PFAS from industrial batch processes might be easily overlooked. To address this deficiency and obtain in-depth information on the occurrence and temporal trend of PFAS in surface water impacted by treated industrial waste water, a comprehensive target and NTA study was implemented for 29 months. Elevated PFAS concentrations with up to 10.8 μg L-1 were detected in the river water by target analysis. In addition to PFAS target analysis, the water samples were analyzed by liquid chromatography-high-resolution tandem mass spectrometry (LC-HRMS/MS). Data processing strategies and various filtering steps were applied to prioritize PFAS. Substances were identified by comparing data to available internal and external PFAS suspect lists, a fragment ion and neutral loss list, and spectral libraries. Several compounds were unequivocally identified based on reference standards. Fifty-five PFAS were (tentatively) identified using NTA. Of those, 43 could be assigned to 13 different homologous series. Partly fluorinated short-chain carboxylic acids (H-PFCA) and sulfonic acids (H-PFSA) were predominantly found in addition to perfluoroalkyl carboxylic acids (PFCA) and the alkyl ether carboxylic acid DONA. To the best of our knowledge, 12 PFAS were reported in surface water for the first time. Signal intensities of individual PFAS and signal ratios varied widely over time, which may indicate batch operations leading to discontinuous emission. Results and insights from this screening approach on PFAS can be used to optimize forthcoming surface water monitoring programs by including newly identified PFAS and selecting appropriate sampling intervals.


Association Between Race/Ethnicity and COVID-19 Outcomes in Systemic Lupus Erythematosus Patients From the United States: Data From the COVID-19 Global Rheumatology Alliance.
To determine the association between race/ethnicity and COVID-19 outcomes in individuals with systemic lupus erythematosus (SLE). Individuals with SLE from the US with data entered into the COVID-19 Global Rheumatology Alliance registry between March 24, 2020 and August 27, 2021 were included. Variables included age, sex, race, and ethnicity (White, Black, Hispanic, other), comorbidities, disease activity, pandemic time period, glucocorticoid dose, antimalarials, and immunosuppressive drug use. The ordinal outcome categories were: not hospitalized, hospitalized with no oxygenation, hospitalized with any ventilation or oxygenation, and death. We constructed ordinal logistic regression models evaluating the relationship between race/ethnicity and COVID-19 severity, adjusting for possible confounders. We included 523 patients; 473 (90.4%) were female and the mean ± SD age was 46.6 ± 14.0 years. A total of 358 patients (74.6%) were not hospitalized; 40 patients (8.3%) were hospitalized without oxygen, 64 patients (13.3%) were hospitalized with any oxygenation, and 18 (3.8%) died. In a multivariable model, Black (odds ratio [OR] 2.73 [95% confidence interval (95% CI) 1.36-5.53]) and Hispanic (OR 2.76 [95% CI 1.34-5.69]) individuals had higher odds of more severe outcomes than White individuals. Black and Hispanic individuals with SLE experienced more severe COVID-19 outcomes, which is consistent with findings in the US general population. These results likely reflect socioeconomic and health disparities and suggest that more aggressive efforts are needed to prevent and treat infection in this population.