Recognizing nursing informatics.
In our climate of economic uncertainty, readily accessible and reliable information is essential for cost effective decision making and forward planning in an efficient and effective manner. Information systems are widely used in the business industries to achieve the above. The state hospitals in South Africa have also realized the need for functional information systems. Where does this leave the division of nursing? This division usually comprises at least one third of the hospital establishment with very specific and often complex needs. The nurse managers at all levels have to plan, predict, forecast, monitor, and control in a similar manner to the business industries. What makes their decisions more critical is that they are dealing with human lives. A nursing informatics section with specially trained nurses will have to become an integral part of nursing, to enable nursing to keep abreast of the increasing demand for accurate and timely information for effective decision making. This paper will evaluate the importance of recognizing the need for an informatics section to evolve within the division of nursing, rather than within the general department of medical informatics.


Health-Promoting Lifestyle in Colorectal Cancer Survivors: A Qualitative Study on the Experiences and Perspectives of Colorectal Cancer Survivors and Healthcare Providers.
Understanding the experiences of survivors and healthcare providers about health-promoting lifestyle (HPL) in colorectal cancer (CRC) survivors is important in planning for coping with the disease, managing treatment side effects, increasing survival, and improving quality of life (QOL). This study was conducted to explore the experiences and perspectives of CRC survivors and healthcare providers about HPL in CRC survivors. This descriptive qualitative study was performed in 2020 at Omid and Imam Reza Hospitals in Mashhad, Iran. Participants were CRC survivors (n = 12) and healthcare providers (n = 33) who were selected by purposive sampling. Data were collected using in-depth semi-structured interview by face to face and then analyzed by Zhang and Wildemuth content analysis method. MaxQDA software was used to organize the data. Following the treatment of cancer, CRC survivors seek to make changes in lifestyle and they choose a HPL that maintains or improves their health. HPL in CRC survivors includes nutrition, activity and rest, health responsibility, interpersonal relations, spiritual growth, and psychological management. The results showed that HPL can lead to motivation, the ability to self-care and improve daily performance, reduce treatment complications, and increase the QOL. CRC survivors can help change their lifestyle patterns with healthy eating, treatment adherence, regular physical activity, and good sleep habits. Furthermore, effective personal and social relationships, spiritual growth, and management of psychological disorders develop health-promoting behaviors in them. CRC survivors also face challenges and limitations in their life after treatment; identifying the components of a HPL in CRC survivors can lead to desirable care, treatment, education, and counseling services.


[Axial lumbar interbody fusion: prospective monocentric study].
The aim of this prospective study was to evaluate clinical and radiographic results in the patients who underwent L5-S1 fixation using the technique of percutaneous lumbar interbody fusion (AxiaLIF). The study comprised 23 patients, 11 women and 12 men, who ranged from age of 21 to 63 years, with an average of 48.2 years. In all patients surgical posterior stabilisation involving the L5-S1 segment had previously been done. The initial indications for surgery were L5-S1 spondylolisthesis in 20 and L5-S1 spondylosis and stenosis in three patients. The AxiaLIF technique for L5-S1 fixation was indicated in overweight patients and in those after repeated abdominal or retroperitoneal surgery. A suitable position and shape of the sacrum or lumbosacral junction was another criterion. The patients were evaluated between 26 and 56 months (average, 40.4 months) after primary surgery and, on the basis of CT and radiographic findings, bone union and lumbosacral junction stability were assessed. The clinical outcome was investigated using the ODI and VAS systems and the results were statistically analysed by the Wilcoxon test for paired samples with statistical significance set at a level of 0.05. The average VAS value was 6.6 before surgery and, after surgery, 5.2 at three months, 4.2 at six months, 3.1 at one year, 2.9 at two years and 2.1 at three years (n=18). At two post-operative years, improvement in the VAS value by 56.1% was recorded. The average pre-operative ODI value was 25.1; the post-operative values were 17.0 at six months, 12.3 at one year, 10.6 at two years and 8.2 at three years (n=18). At two years after surgery the ODI value improved by 57.8%. To the question concerning their willingness to undergo, with acquired experience, surgery for the same diagnosis, 21 patients (91.3%) gave an affirmative answer. Neither screw breakage nor neurovascular damage or rectal injury was found. CT scans showed complete interbody bone fusion in 22 of the 23 patients (95.6%), In one patient the finding was not clear. Also, posterolateral fusion was achieved in all but one patients (95.6%). A stable L5-S1 segment was found in all patients at all follow-up intervals. The improvement in both VAS and ODI values was statistically significant. In addition to indications usual in degenerative disc disease, overweight patients, those who had repeated trans- or retroperitoneal surgery in the L5-S1 region or who underwent long posterior fixation to stabilise the caudal margin of instrumentation are indicated for the AxiaLIF procedure. The clinical results of our study are in agreement with the conclusions of other studies and are similar to the outcomes of surgery using other types of fusion or dynamic stabilisation for this diagnosis. The high rate of fusion in our group is affected by use of a rigid transpedicular fixator together with posterolateral arthrodesis. On the other hand, no negative effects of only synthetic bone applied to interbody space were recorded. The percutaneous axial pre-sacral approach to the L5-S1 interbody space with application of a double-treaded screw is another option for the management of this much strained segment. The technique is useful particularly when contraindications for conventional surgical procedures are present in patients with anatomical anomalies, in overweight patients or in those who have had repeated surgery in the region. Clinical outcomes and the success rate for L5-S1 bone fusion are comparable with conventional techniques. Complications are rare but their treatment is difficult.


A case of isolated plexiform neurofibroma in a patient with myasthenia gravis.
We report a case of an isolated plexiform neurofibroma occurring in a patient with myasthenia gravis. A 48-year-old man presented with asymptomatic skin-colored nodules on the tip of his 4th finger. Microscopically, a plexiform neurofibroma was identified located in the dermis that appeared to originate from small superficial nerves. He had a 20-year history of treated myasthenia gravis; otherwise, his personal and family histories were unremarkable. Given that myasthenia gravis is a disorder of the peripheral nerves, plexiform neurofibromas could be associated with myasthenia gravis. However, the development of an isolated plexiform neurofibroma in a case of myasthenia gravis has not yet been reported. The occurrence of a neurofibromas in a patient with myasthenia gravis suggests a link in the pathogenesis of these two diseases.


Asbestos removal, health hazards, and the EPA. Council on Scientific Affairs, American Medical Association.
Resolution 193 (A-90), which was adopted by the House of Delegates of the American Medical Association, called on the Council on Scientific Affairs to study the situation regarding asbestos abatement, the risks to health, and the appropriateness of Environmental Protection Agency regulations, policies, and control measures. This report reviews the current status of asbestos abatement as applied to schools and public buildings, which currently accounts for the major expenditure of public funds.


Advances in umbilical cord blood transplant: an overview of the 12th International Cord Blood Symposium, San Francisco, 5-7 June 2014.
From 5 to 7 June the 12th Annual International Cord Blood Symposium was held in San Francisco. The meeting was devoted to advances in umbilical cord blood research with a major focus on translational and clinical results in cord blood transplant and in regenerative medicine. Over 3 days, a comprehensive summary of the state of the art was provided. We have summarized the most important data, organized around the following themes: use of umbilical cord blood for tissue repair, new indications for umbilical cord blood unit stem cell transplant (CBU SCT), enhancing count recovery after CBU SCT, improving outcomes, product quality and financial and cost considerations.


Modulation of the endocrine and immune systems by well-controlled hyperthermia equipment.
Since high levels of hyperthermia induce immunosuppression to a certain extent (i.e., granulocytosis and lymphocytopenia) in patients, we applied mild hyperthermia in volunteers using equipment enabling well-controlled hyperthermia. Restricted control of rectal temperature at 39.4 (+/- 0.2) degrees C for 30 min was conducted and various parameters of the body were examined. The most prominent change observed during exposure to hyperthermia was elevated levels of pH and PO(2) in the blood, even in the venous blood. A transient elevation of ACTH, cortisol and growth hormone in the blood was also seen during this time. In parallel with this phenomenon, the number of total lymphocytes and those of its subsets (especially CD57(+) or CD56(+) NK cells and NKT cells) increased. More interestingly, the proportion of HLA-DR (MHC class II antigens) increased in NK and NKT cells, and their intensity on the surface of CD20(+) B cells increased. These results suggest that mild hyperthermia is important for modulation of the functions of the circulatory, endocrine and immune systems.


High-quality-factor Bragg onion resonators with omnidirectional reflector cladding.
We propose to approximate a spherically symmetric Bragg resonator in an onionlike geometry. We develop a transfer-matrix theory for calculation of the quality factors and analyze the effect of the onion stem on cavity Q factors. We find that it is possible to achieve significant inhibition of spontaneous emission in an onion resonator with omnidirectional cladding layers.


MR findings of spontaneous intracranial hypotension.
To present the MR features of spontaneous intracranial hypotension (SIH) and to discuss the correlation of MR features with clinical manifestations. Between 1997 and 2000, 15 patients with SIH underwent brain MR investigation. Lumbar puncture for the measurement of cerebrospinal fluid (CSF) pressure was performed in 6 patients. Follow-up MR was obtained in 8 patients after the clinical symptoms had improved. We analyzed the MR findings of SIH, and discuss them in relation to CSF pressure and clinical manifestations. Fourteen of the 15 patients with SIH had abnormal MR findings. The imaging findings included: diffuse pachymeningeal enhancement in 13 patients; descent of the brain in 6 patients; and subdural effusion/hematoma in 2 patients. Low CSF pressure was shown at lumbar puncture in 5 out of 6 patients. Follow-up MR examination in the symptoms-relieved status demonstrated a recovery of the abnormal MR findings in 6 of 8 patients. Our data revealed that MR is sensitive in diagnosing SIH. The presence of characteristic clinical manifestations and MR features are virtually diagnostic for SIH syndrome. Invasive lumbar puncture is thus reserved for the equivocal cases.


CD24 controls expansion and persistence of autoreactive T cells in the central nervous system during experimental autoimmune encephalomyelitis.
In the development of experimental autoimmune encephalomyelitis (EAE), a model for multiple sclerosis (MS), autoreactive T cells must be activated and clonally expand in the lymphoid organs, and then migrate into the central nervous system (CNS) where they undergo further activation. It is unclear whether the autoreactive T cells further expand in the CNS and if so, what interactions are required for this process. We have demonstrated previously that expression by the host cells of the heat-stable antigen (CD24), which was recently identified as a genetic modifier for MS, is essential for their susceptibility to EAE. Here we show that CD24 is essential for local clonal expansion and persistence of T cells after their migration into the CNS, and that expression of CD24 on either hematopoietic cells or nonhematopoietic antigen-presenting cells in the recipient is sufficient to confer susceptibility to EAE.


The effect of patient-managed stress dosing on electrolytes and blood pressure in acute illness in children with adrenal insufficiency.
Adrenal crises (AC) are acute episodes of adrenal insufficiency (AI). Manifestations include hypotension and electrolyte disturbances. Glucocorticoid stress dosing (SD) can prevent AC progression, but its effect on physiological parameters has not been assessed in a 'real world setting'. To assess the effect of prior self-managed glucocorticoid dose escalation on physiological markers in children with congenital adrenal hyperplasia (CAH) presenting to hospital for an acute illness. An audit of records of all children with CAH presenting to paediatric referral hospital between 2000 and 2015. Potassium, sodium and glucose levels, and hypotension were compared between children who had and had not used SD. There were 321 attendances by patients with CAH and an acute illness during the study period. Any form of SD was used by 64.2% (n = 206); intramuscular (IM) hydrocortisone was used by 22.1% (n = 71) and oral only by 41.7% (n = 134). Use of SD (oral and/or IM) was associated with a significantly lower mean potassium level (4.02 ± 0.71 vs. 4.27 ± 0.79 mmol/l, P < .05). Linear regression analysis showed that age (beta: -0.04 years (95% CI -0.06, -0.02)), diarrhoea (beta: -0.41 (95% CI -0.06, -0.02)) and any form of stress dosing (oral, IM or both) (beta: -0.29 (95% CI -0.55, -0.04)) were each independently and significantly associated with potassium levels. SD was not significantly associated with sodium or glucose concentrations or with estimates of hypotension. Patient-initiated SD resulted in a significant reduction in hyperkalaemia and lowered mean potassium levels in paediatric patients with CAH but did not alter significantly sodium and glucose concentrations or incidences of hypotension.


Toll-Like Receptor-4 Inhibitor TAK-242 Attenuates Motor Dysfunction and Spinal Cord Pathology in an Amyotrophic Lateral Sclerosis Mouse Model.
Neuroinflammation contributes to amyotrophic lateral sclerosis (ALS) progression. TLR4, a transmembrane protein that plays a central role in activation of the innate immune system, has been shown to induce microglial activation in ALS models. TLR4 is up-regulated in the spinal cords of hSOD1G93A mice. We aimed to examine the effects of specific TLR4 inhibition on disease progression and survival in the hSOD1G93A mouse model of ALS. Immunologic effect of TLR4 inhibition in vitro was measured by the effect of TAK-242 treatment on LPS-induced splenocytes proliferation. hSOD1G93A transgenic mice were treated with TAK-242, a selective TLR4 inhibitor, or vehicle. Survival, body weight, and motor behavior were monitored. To evaluate in vivo immunologic modifications associated with TAK-242 treatment, we measured serum IL-1β in the plasma, as well as IL-1β and TNF-α mRNAs in the spinal cord in wild-type mice and in TAK-242-treated and vehicle-treated early symptomatic hSOD1G93A mice. Immunohistochemical analysis of motor neurons, astrocytes, and microglial reactivity in the spinal cords were performed on symptomatic (100 days old) TAK-242-treated and vehicle-treated hSOD1G93A mice. In vitro, splenocytes taken from 100 days old hSOD1G93A mice showed significantly increased proliferation when exposed to LPS (p = 0.0002), a phenomenon that was reduced by TAK-242 (p = 0.0179). TAK-242 treatment did not attenuate body weight loss or significantly affect survival. However, TAK-242-treated hSOD1G93A mice showed temporary clinical delay in disease progression evident in the ladder test and hindlimb reflex measurements. Plasma IL-1β levels were significantly reduced in TAK-242-treated compared to vehicle-treated hSOD1G93A mice (p = 0.0023). TAK-242 treatment reduced spinal cord astrogliosis and microglial activation and significantly attenuated spinal cord motor neuron loss at early disease stage (p = 0.0259). Compared to wild-type animals, both IL-1β and TNF-α mRNAs were significantly upregulated in the spinal cords of hSOD1G93A mice. Spinal cord analysis in TAK-242-treated hSOD1G93A mice revealed significant attenuation of TNF-α mRNA (p = 0.0431), but no change in IL-1β mRNA. TLR4 inhibition delayed disease progression, attenuated spinal cord astroglial and microglial reaction, and reduced spinal motor neuron loss in the ALS hSOD1G93A mouse model. However, this effect did not result in increased survival. To our knowledge, this is the first report on TAK-242 treatment in a neurodegenerative disease model. Further studies are warranted to assess TLR4 as a therapeutic target in ALS.


Mineralisation of reconstituted collagen using polyvinylphosphonic acid/polyacrylic acid templating matrix protein analogues in the presence of calcium, phosphate and hydroxyl ions.
The complex morphologies of mineralised collagen fibrils are regulated through interactions between the collagen matrix and non-collagenous extracellular proteins. In the present study, polyvinylphosphonic acid, a biomimetic analogue of matrix phosphoproteins, was synthesised and confirmed with FTIR and NMR. Biomimetic mineralisation of reconstituted collagen fibrils devoid of natural non-collagenous proteins was demonstrated with TEM using a Portland cement-containing resin composite and a phosphate-containing fluid in the presence of polyacrylic acid as sequestration, and polyvinylphosphonic acid as templating matrix protein analogues. In the presence of these dual biomimetic analogues in the mineralisation medium, intrafibrillar and extrafibrillar mineralisation via bottom-up nanoparticle assembly based on the non-classical crystallisation pathway could be identified. Conversely, only large mineral spheres with no preferred association with collagen fibrils were observed in the absence of biomimetic analogues in the medium. Mineral phases were evident within the collagen fibrils as early as 4 h after the initially-formed amorphous calcium phosphate nanoprecursors were transformed into apatite nanocrystals. Selected area electron diffraction patterns of highly mineralised collagen fibrils were nearly identical to those of natural bone, with apatite crystallites preferentially aligned along the collagen fibril axes.


[Method of quantitative evaluation of the reactivity of the microvessels to vasoactive substances in patients with hypertensive disease].
A method developed by the authors is described that consists in a quantitative test for the evaluation of the reactivity of the microvessels judging by the degree of changes in the resorption speed of isotopes from a tissue depot. In patients with essential hypertension an increased vascular reactivity was revealed with reference to angiotensin, noradrenalin and serotonin, as well as an increased duration of the effect of angiotensin. The highest vascular reactivity was noted in patients with stage II hypertension who have retained a hyperkinetic type of circulation.


Indicators for the evaluation of diet quality.
The role of diet quality and physical activity in reducing the progression of chronic disease is becoming increasingly important. Dietary Quality Indices or Indicators (DQIs) are algorithms aiming to evaluate the overall diet and categorize individuals according to the extent to which their eating behaviour is "healthy". Predefined indexes assess dietary patterns based on current nutrition knowledge and they have been developed primarily for nutritional epidemiology to assess dietary risk factors for non-communicable diseases. There are many different types of DQIs. There are three major categories of DQIs: a) nutrient-based indicators; b) food/food group based indicators; and c) combination indexes, the vast majority of DQIs, which often include a measure of diet variety within and across food groups, a measure of adequacy i.e. nutrients (compared to requirements) or food groups (quantities or servings), a measure of nutrients/foods to consume in moderation, and an overall balance of macronutrients. The Healthy Eating Index (HEI), the Diet Quality Index (DQI), the Healthy Diet Indicator (HDI) and the Mediterranean Diet Score (MDS) are the four 'original' diet quality scores that have been referred to and validated most extensively. Several indexes have been adapted and modified from those originals. In particular, many variations on the MDS have been proposed, included different alternate MDS and Mediterranean Diet Adherence Screener (MEDAS). Primary data source of DQI's are individual dietary data collection tools, namely 24 h quantitative intake recalls, dietary records and food frequency questionnaires. Nutrients found in many scores are total fat, saturated fatty acids or the ratio of monounsaturated fatty acids to saturated fatty acids or the latter SFA to polyunsaturated fatty acids. Cholesterol, protein content and quality, complex carbohydrates, mono- and disaccharides, dietary fibre and sodium are also found in various scores. All DQIs, except those that only contain nutrients, include the components fruits and vegetables; additional attributes are legumes or pulses, nuts and seeds. Meat and meat products, namely red and processed meat, poultry, and milk and dairy products are also included in many scores. Other foods contained in some DQIs e.g. MDS are olive oil and fish. Nowadays, there is interest in defining more than DQIs, healthy life indices (HLIs), which give information on behaviours associated with specific patterns and beyond dietary habits they include physical activity, rest and selected socio-cultural habits. The Mediterranean Lifestyle (MEDLIFE) index has been recently created based on the current Spanish Mediterranean food guide pyramid and it includes both the assessment of food consumption directly related to the Mediterranean diet, physical activity and rest and other relevant cultural information. However, a global HLI should consider, based on the Iberoamerican Nutrition Foundation (FINUT) Pyramid of Healthy Lifestyles, in addition to food groups and nutrients, selected items on food safety e.g. consumption rate of proceed foods, food handling, preparation and storage and access to drinking water, selected food habits, including alcoholic beverage and salt consumption patterns, purchase of seasonal and local foods, home cooking and conviviality, as well as patterns of physical activity, sedentary and rest habits and some selected sociocultural habits, particularly those related to food selection, religious beliefs and socializing with friends.


Can the impacts of cold-water pollution on fish be mitigated by thermal plasticity?
Increasingly, cold-water pollution (CWP) is being recognised as a significant threat to aquatic communities downstream of large, bottom-release dams. Cold water releases typically occur during summer when storage dams release unseasonably cold and anoxic hypolimnetic waters, which can decrease the temperature of downstream waters by up to 16°C. Depending on the release duration, these hypothermic conditions can persist for many months. The capacity of ectothermic species to tolerate or rapidly adjust to acute temperature changes may determine the nature and magnitude of the impact of CWP on affected species. This study assessed the impacts of an acute reduction in water temperature on the physiological function and locomotor performance of juvenile silver perch (Bidyanus bidyanus) and examined their capacity to thermally compensate for the depressive effects of low temperatures via phenotypic plasticity. Locomotor performance (Ucrit and Usprint) and energetic costs (routine and maximum metabolic rate) were measured at multiple points over a 10-week period following an abrupt 10°C drop in water temperature. We also measured the thermal sensitivity of metabolic enzymes from muscle samples taken from fish following the exposure period. Cold exposure had significant depressive effects on physiological traits, resulting in decreases in performance between 10% and 55%. Although there was partial acclimation of Ucrit (~35% increase in performance) and complete compensation of metabolic rate, this occurred late in the exposure period, meaning silver perch were unable to rapidly compensate for the depressive effects of thermal pollution. The results of this study have substantial implications for the management of cold water releases from large-scale dams and the conservation of native freshwater fish species, as this form of thermal pollution can act as a barrier to fish movement, cause reduced recruitment, ecological community shifts and disruptions to timing and success of reproduction.


The impact of life course exposures to neighbourhood deprivation on health and well-being: a review of the long-term neighbourhood effects literature.
In this review article, we detail a small but growing literature in the field of health geography that uses longitudinal data to determine a life course component to the neighbourhood effects thesis. For too long, there has been reliance on cross-sectional data to test the hypothesis that where you live has an effect on your health and well-being over and above your individual circumstances. We identified 53 articles that demonstrate how neighbourhood deprivation measured at least 15 years prior affects health and well-being later in life using the databases Scopus and Web of Science. We find a bias towards US studies, the most common being the Panel Study of Income Dynamics. Definition of neighbourhood and operationalization of neighbourhood deprivation across most of the included articles relied on data availability rather than a priori hypothesis. To further progress neighbourhood effects research, we suggest that more data linkage to longitudinal datasets is required beyond the narrow list identified in this review. The limited literature published to date suggests an accumulation of exposure to neighbourhood deprivation over the life course is damaging to later life health, which indicates improving neighbourhoods as early in life as possible would have the greatest public health improvement.


Acceptability and appropriateness of a risk-tailored organised melanoma screening program: Qualitative interviews with key informants.
In Australia, opportunistic screening (occurring as skin checks) for the early detection of melanoma is common, and overdiagnosis is a recognised concern. Risk-tailored cancer screening is an approach to cancer control that aims to provide personalised screening tailored to individual risk. This study aimed to explore the views of key informants in Australia on the acceptability and appropriateness of risk-tailored organised screening for melanoma, and to identify barriers, facilitators and strategies to inform potential future implementation. Acceptability and appropriateness are crucial, as successful implementation will require a change of practice for clinicians and consumers. This was a qualitative study using semi-structured interviews. Key informants were purposively selected to ensure expertise in melanoma early detection and screening, prioritising senior or executive perspectives. Consumers were expert representatives. Data were analysed deductively using the Tailored Implementation for Chronic Diseases (TICD) checklist. Thirty-six participants were interviewed (10 policy makers; 9 consumers; 10 health professionals; 7 researchers). Key informants perceived risk-tailored screening for melanoma to be acceptable and appropriate in principle. Barriers to implementation included lack of trial data, reluctance for low-risk groups to not screen, variable skill level in general practice, differing views on who to conduct screening tests, confusing public health messaging, and competing health costs. Key facilitators included the perceived opportunity to improve health equity and the potential cost-effectiveness of a risk-tailored screening approach. A range of implementation strategies were identified including strengthening the evidence for cost-effectiveness, engaging stakeholders, developing pathways for people at low risk, evaluating different risk assessment criteria and screening delivery models and targeted public messaging. Key informants were supportive in principle of risk-tailored melanoma screening, highlighting important next steps. Considerations around risk assessment, policy and modelling the costs of current verses future approaches will help inform possible future implementation of risk-tailored population screening for melanoma.


Identification of large genetic variations in the equine infectious anemia virus tat-gag genomic region.
The aetiological agent of equine infectious anaemia (EIA) is the retrovirus equine infectious anemia virus (EIAV) that infects all members of the Equidae family. The EIA is widely disseminated in the Brazilian territory with a high seroprevalence in the Brazilian Pantanal and is mainly diagnosed using agar gel immunodiffusion (AGID). There are few complete EIAV genome sequences available in GenBank, which had an impact on molecular detection studies. In this study, we conducted molecular detection and sequencing of EIAV proviral DNA from Brazilian horses. We analysed the genomic region from exon 1 of tat to gag (tat-gag). Comparative serological tests, comprising AGID and two enzyme-linked immunosorbent assays (ELISAs), were also conducted. Of the 133 samples, 58 were positive in the tat-gag PCR, and 49 nucleotide sequences of 272 bp were obtained. Using this developed tat-gag PCR EIAV proviral DNA was detected in 7% of the AGID-negative samples and 26% of the AGID-negative samples were positive in at least one of the ELISA tests used. Using phylogenetic analysis, the Brazilian Pantanal EIAV sequences grouped in a different clade of EIAV sequences from other countries. Thus, the EIAV sequences can contribute to the knowledge of the tat-gag genomic region in the circulating viruses in the Brazilian Pantanal, in addition to providing new information about the genetic diversity. In addition, the serological results demonstrate the greater sensitivity of the ELISAs used in this study compared to AGID for EIA diagnosis.


Role of renin-angiotensin system in hypotension-evoked thirst: studies with hydralazine.
Injection of rats either with diazoxide (25 mg/kg iv), isoproterenol (0.33 mg/kg sc), or hydralazine (HDZ) (10 mg/kg ip) decreased arterial blood pressure from approximately 120 to 70-80 mmHg and stimulated renin secretion. However, diazoxide and isoproterenol treatments each stimulated water ingestion, whereas HDZ treatment did not. HDZ treatment did not reduce water intake evoked by systemic injection of hypertonic saline or 20% polyethylene glycol solution or by 24-h water deprivation, suggesting that HDZ treatment did not interfere with drinking behavior. In contrast, HDZ treatment markedly reduced water intake evoked by injection of diazoxide or isoproterenol or by intravenous infusion of renin. Furthermore, a highly significant correlation was observed when plasma ANG II levels were plotted as a function of plasma renin activity after intravenous infusion of renin and after diazoxide and isoproterenol treatments. However, values obtained after HDZ treatment alone or in combination with intravenous infusion of renin did not fall near the 99% confidence interval of the regression line, suggesting that HDZ treatment blocks ANG II production and/or promotes its clearance. Thus rats apparently do not increase water intake after HDZ treatment, because this drug interferes with the renin-angiotensin system. These results provide further evidence that arterial hypotension evokes thirst in rats predominantly by activation of the renin-angiotensin system.


Differentiation of Motor Neuron-Like Cells from Tonsil-Derived Mesenchymal Stem Cells and Their Possible Application to Neuromuscular Junction Formation.
Human tonsil-derived mesenchymal stem cells (T-MSCs) are newly identified MSCs and present typical features of MSCs, including having the differentiation capacity into the three germ layers and excellent proliferation capacity. They are easily sourced and are useful for stem cell therapy in various disease states. We previously reported that T-MSCs could be differentiated into skeletal myocytes and Schwann-like cells; therefore, they are a promising candidate for cell therapies for neuromuscular disease. Motor neurons (MNs), which regulate spontaneous behavior, are affected by a wide range of MN diseases (MNDs) for which there are no effective remedies. We investigated the differentiation potential of MN-like cells derived from T-MSCs (T-MSC-MNCs) for application to therapy of MNDs. After the process of MN differentiation, the expression of MN-related markers, including Islet 1, HB9/HLXB9 (HB9), and choline acetyltransferase (ChAT), was increased when compared with undifferentiated T-MSCs. The secretion of acetylcholine to the conditioned medium was significantly increased after MN differentiation. We cocultured T-MSC-MNCs and human skeletal muscle cells, and confirmed the presence of the acetylcholine receptor clusters, which demonstrated the formation of neuromuscular junctions. The potential functional improvements afforded by these T-MSC-MNCs could be useful in the treatment of MNDs caused by genetic mutation, viral infection, or environmental problems.


Identification and analysis of function of a novel splicing variant of mouse receptor activator of NF-κB.
Receptor activator of NF-κB (RANK) is a member of the tumor necrosis factor receptor (TNFR) family expressed in osteoclast precursors, and RANK-RANK ligand (RANKL) signaling is a key system for differentiation, activation and survival of osteoclasts. Here, we report the identification of a novel alternative splicing variant of mouse RANK gene (vRANK) that contains a new intervening exon between exon 1 and exon 2 of mouse full-length RANK (fRANK) mRNA. Since this novel exon contains the stop codon, vRANK encodes truncated amino acids that have a portion of the signal peptide of fRANK and an additional 19 amino acids that show no homology to previously reported domains. By transient transfection studies with vRANK-GFP and -Flag expressing constructs, vRANK was found localized mostly in the cytoplasm and partly in the cell membrane, but was not secreted into the culture supernatant. Under the stimulation of various factors, the expression of vRANK mRNA was almost parallel to that of fRANK in RAW264.7 cells not treated with M-CSF. Overexpression of vRANK, on the other hand, decreased TRACP (a marker of osteoclasts) mRNA expression as well as the number of TRACP-positive multinucleated giant cells. While the mRNA expression levels of NFATc1 (a master transcriptional factor of the osteoclast differentiation program) were not affected, apoptotic cells increased significantly in vRAN K-transfected cells treated with sRANKL. Taken together, these results suggest that vRANK is a novel osteoclast suppressor that reduces the number of RANKL-induced mature osteoclasts mainly by negating the anti-apoptotic effect of RANKL.


Effects of D-cycloserine on craving to alcohol cues in problem drinkers: preliminary findings.
It has been shown that the partial N-methyl-d-aspartate (NMDA) agonist d-cycloserine (DCS) facilitates exposure-based learning in humans with anxiety disorders. However, the effects of DCS on exposure to substance cues are still uncertain. The primary aim of the study was to examine the effects of DCS on exposure sessions to alcohol cues. Twenty non-treatment-seeking problem drinkers were randomly assigned to receive 50 mg of DCS or placebo 50 minutes prior to each of 3 alcohol cue exposure sessions consisting of 5 exposure trials within an 8-day period. Following this, participants underwent two test sessions without the administration of any medication. The test sessions occurred 3 and 7 days after the last cue exposure session, respectively. Dependent measures included drinking urge and heart rate and drinking urge during the test sessions. Individuals who received DCS showed increased craving to alcohol cues as compared with individuals who received placebo during the first test session. No group difference in drinking urge was found during the second test session. Furthermore, the groups did not differ in heart rate at any of the assessment points. The results suggest that DCS may temporarily augment cue-elicited craving for alcohol. As in an earlier study with cocaine-dependent individuals, DCS appears to exhibit a different profile in problem drinkers to those with anxiety disorders. Dose, timing, arousal, and treatment motivation as considerations are discussed, as are methodological considerations and the need for additional studies in this area.


The gene encoding the transcription factor SCIP has features of an expressed retroposon.
SCIP is a POU domain transcription factor expressed by glial progenitor cells in the peripheral and central nervous systems (dividing Schwann cells and O-2A cells, respectively), where it appears to act as a repressor of myelin-specific genes. We have isolated genomic clones encoding the rat SCIP gene. Comparison of the structure of these clones with genomic Southern blots and SCIP cDNAs demonstrates that SCIP is encoded in a single-copy, intronless gene that has the general features of an expressed retroposon. This gene contributes to an extended CpG island. It is transcribed to produce a 3.1-kb mRNA that encodes a 451-amino-acid protein with a predicted molecular mass of 45 kDa. Immunopurified SCIP antibodies specifically recognize a nuclear protein of this size in cultured proliferating Schwann cells, and gel shift analyses demonstrate that this protein is the predominant octamer-binding protein in these cells.


Ions and IVs: are we still using the Henry Ford model?
Intravenous fluid therapy matched to fluid losses provides the patient with appropriate and cost-effective care. While the physiognomies of many of our patients are robust enough to handle nearly any type of intravenous perioperative replacement, we shall all see patients with varying fluid and electrolyte disturbances. Using the tables and formulae in this article will assist the anesthesia provider in tailoring fluids to both tonicity and electrolyte needs.


Pathological Significance and Prognostic Value of Surfactant Protein D in Cancer.
Surfactant protein D (SP-D) is a pattern recognition molecule belonging to the Collectin (collagen-containing C-type lectin) family that has pulmonary as well as extra-pulmonary existence. In the lungs, it is a well-established opsonin that can agglutinate a range of microbes, and enhance their clearance via phagocytosis and super-oxidative burst. It can interfere with allergen-IgE interaction and suppress basophil and mast cell activation. However, it is now becoming evident that SP-D is likely to be an innate immune surveillance molecule against tumor development. SP-D has been shown to induce apoptosis in sensitized eosinophils derived from allergic patients and a leukemic cell line via p53 pathway. Recently, SP-D has been shown to suppress lung cancer progression via interference with the epidermal growth factor signaling. In addition, a truncated form of recombinant human SP-D has been reported to induce apoptosis in pancreatic adenocarcinoma via Fas-mediated pathway in a p53-independent manner. To further establish a correlation between SP-D presence/levels and normal and cancer tissues, we performed a bioinformatics analysis, using Oncomine dataset and the survival analysis platforms Kaplan-Meier plotter, to assess if SP-D can serve as a potential prognostic marker for human lung cancer, in addition to human gastric, breast, and ovarian cancers. We also analyzed immunohistochemically the presence of SP-D in normal and tumor human tissues. We conclude that (1) in the lung, gastric, and breast cancers, there is a lower expression of SP-D than normal tissues; (2) in ovarian cancer, there is a higher expression of SP-D than normal tissue; and (3) in lung cancer, the presence of SP-D could be associated with a favorable prognosis. On the contrary, at non-pulmonary sites such as gastric, breast, and ovarian cancers, the presence of SP-D could be associated with unfavorable prognosis. Correlation between the levels of SP-D and overall survival requires further investigation. Our analysis involves a large number of dataset; therefore, any trend observed is reliable. Despite apparent complexity within the results, it is evident that cancer tissues that produce less levels of SP-D compared to their normal tissue counterparts are probably less susceptible to SP-D-mediated immune surveillance mechanisms via infiltrating immune cells.


3D indoor modeling and game theory based navigation for pre and post COVID-19 situation.
The COVID-19 pandemic has greatly affected human behavior, creating a need for individuals to be more cautious about health and safety protocols. People are becoming more aware of their surroundings and the importance of minimizing the risk of exposure to potential sources of infection. This shift in mindset is particularly important in indoor environments, especially hospitals, where there is a greater risk of virus transmission. The implementation of route planning in these areas, aimed at minimizing interaction and exposure, is crucial for positively influencing individual behavior. Accurate maps of buildings help provide location-based services, prepare for emergencies, and manage infrastructural facilities. There aren't any maps available for most installations, and there are no proven techniques to categorize features within indoor areas to provide location-based services. During a pandemic like COVID-19, the direct connection between the masses is one of the significant preventive steps. Hospitals are the main stakeholders in managing such situations. This study presents a novel method to create an adaptive 3D model of an indoor space to be used for localization and routing purposes. The proposed method infuses LiDAR-based data-driven methodology with a Quantum Geographic Information System (QGIS) model-driven process using game theory. The game theory determines the object localization and optimal path for COVID-19 patients in a real-time scenario using Nash equilibrium. Using the proposed method, comprehensive simulations and model experiments were done using QGIS to identify an optimized route. Dijkstra algorithm is used to determine the path assessment score after obtaining several path plans using dynamic programming. Additionally, Game theory generates path ordering based on the custom scenarios and user preference in the input path. In comparison to other approaches, the suggested way can minimize time and avoid congestion. It is demonstrated that the suggested technique satisfies the actual technical requirements in real-time. As we look forward to the post-COVID era, the tactics and insights gained during the pandemic hold significant value. The techniques used to improve indoor navigation and reduce interpersonal contact within healthcare facilities can be applied to maintain a continued emphasis on safety, hygiene, and effective space management in the long term. The use of three-dimensional (3D) modeling and optimization methodologies in the long-term planning and design of indoor spaces promotes resilience and flexibility, encouraging the adoption of sustainable and safe practices that extend beyond the current pandemic.


High opportunity for postcopulatory sexual selection under field conditions.
In polygamous systems, male fitness is determined not only by mating success but also by fertilization success. Despite the growing interest over the past several decades in postcopulatory sexual selection, its relative importance compared to precopulatory sexual selection remains a subject of debate. Here, we use extensive behavioral observations of a seminatural population of Hawaiian swordtail crickets, Laupala cerasina, and molecular paternity assignment to measure the opportunities for pre- and postcopulatory selection. Because postcopulatory selection has the potential to operate at multiple stages, we also separately attribute its effects to factors specific to mating events versus factors specific to males. We find that variance in postcopulatory success is over four times as great as variance in precopulatory success, with most of it unexplained by male mating order or the number of nuptial gifts given. Surprisingly, we also find that male singing effort is under postcopulatory selection, suggesting that males who sing more frequently also have more competitive ejaculates. Our results are consistent with the hypothesis that high polyandry levels promote greater relative postcopulatory selection. They also highlight the need for detailed behavioral observations under conditions as natural as possible when measuring mating and reproductive success.


The chromatin-modifying enzyme Ezh2 is critical for the maintenance of regulatory T cell identity after activation.
Regulatory T cells (Treg cells) are required for immune homeostasis. Chromatin remodeling is essential for establishing diverse cellular identities, but how the epigenetic program in Treg cells is maintained throughout the dynamic activation process remains unclear. Here we have shown that CD28 co-stimulation, an extracellular cue intrinsically required for Treg cell maintenance, induced the chromatin-modifying enzyme, Ezh2. Treg-specific ablation of Ezh2 resulted in spontaneous autoimmunity with reduced Foxp3(+) cells in non-lymphoid tissues and impaired resolution of experimental autoimmune encephalomyelitis. Utilizing a model designed to selectively deplete wild-type Treg cells in adult mice co-populated with Ezh2-deficient Treg cells, Ezh2-deficient cells were destabilized and failed to prevent autoimmunity. After activation, the transcriptome of Ezh2-deficient Treg cells was disrupted, with altered expression of Treg cell lineage genes in a pattern similar to Foxp3-deficient Treg cells. These studies reveal a critical role for Ezh2 in the maintenance of Treg cell identity during cellular activation.


Predictors of subjective age before and after cataract surgery: conscientiousness makes a difference.
This study contributes to research on subjective age by examining the interplay of felt age with health, functional limitations, and personality. Individuals undergoing cataract surgery (N=134; 38 to 92 years of age) provided data for an assessment period of 6 weeks surrounding their scheduled surgery. Conscientiousness and repeated measurements of health indicators, functional limitations, and felt age were included in the analyses. Results indicated that functional limitations may be more important to the construction of felt age than their underlying health-related causes. Moreover, conscientious participants felt younger before and after surgery. Their functional status was less dependent on health than the functional status of less conscientious participants. This moderator effect is discussed along with health-related pathways leading to felt age.


Gross motor function is an important predictor of daily physical activity in young people with bilateral spastic cerebral palsy.
The aim of the study was to describe daily physical activity levels of adolescents and young adults with bilateral spastic cerebral palsy (CP) and to identify factors that help predict these levels. Daily physical activity was measured using an accelerometer-based activity monitor in 45 young people with bilateral spastic CP (23 males, 22 females; mean age 18y 6mo [SD 2y 5mo] range 16y 1mo-20y 11mo); classified as Gross Motor Function Classification System (GMFCS) level II or III and with contractures of <20° at hip and knee. Predictor variables included demographic characteristics (age, sex, weight) and physical characteristics (gross motor function, lower limb muscle strength, 6min walk distance). Data were analyzed using the information-theoretic approach, using the Akaike information criterion (AIC) and linear regression. Daily activity levels were low compared with published norms. Gross Motor Function Measure Dimension-E (GMFM-E; walking, running, and jumping) was the only common predictor variable in models that best predicted energy expenditure, number of steps, and time spent sitting/lying. GMFM Dimension-D (standing) and bilateral reverse leg press strength contributed to the models that predicted daily physical activity. Adolescents and young adults with bilateral spastic CP and mild to moderate walking disabilities have low levels of daily activity. The GMFM-E was an important predictor of daily physical activity.


Extended-Spectrum Beta-Lactamase-/AmpC Beta-Lactamase-Producing Enterobacteriaceae in Broiler Farms: Transmission Dynamics at Farm Level.
The occurrence of extended-spectrum beta-lactamase- (ESBL) and/or AmpC beta-lactamase- (AmpC) producing Enterobacteriaceae in livestock, especially in broiler fattening flocks, has been demonstrated in previous studies. Nevertheless, data on transmission routes of these resistant bacteria into the fattening farms are rare. Therefore, seven broiler fattening flocks were investigated for the occurrence of ESBL-/AmpC-producing Enterobacteriaceae during the course of the fattening period with the special focus on horizontal transmission routes. ESBL-/AmpC-producing Enterobacteriaceae from both individual animals and their housing environment were isolated at different time points and the housing environment was even sampled before the arrival of the chickens. All obtained ESBL-/AmpC-producing Enterobacteriaceae were examined for their bacterial species, Escherichia coli phylogroup, and occurrence of resistance genes. Selected isolates were further analyzed via whole-genome sequencing. All seven investigated flocks were tested positive for ESBL-/AmpC-producing Enterobacteriaceae with widely varying prevalence between the flocks. In one flock, the ESBL-/AmpC-producing Enterobacteriaceae were already detected in the housing environment before the arrival of the animals. In general, among the different types of ESBL-/AmpC-producing Enterobacteriaceae determined E. coli harboring a blaCMY-2 gene was the most frequent. Using whole-genome analyses we observed a horizontal transmission of ESBL-/AmpC-producing Enterobacteriaceae through contaminated housing environment as two flocks consecutively fattened in the same farm harbored closely related ESBL-producing isolates. This demonstrates the influence of a previous fattened flock on the ESBL-/AmpC-status of a following broiler flock and, therefore, the importance of hygiene measures on farm level.


Clinical factors associated with autologous stem cell transplantation outcomes in multiple myeloma: upfront transplant with MEL200 remains the standard of care.
Autologous stem cell transplantation (ASCT) remains the mainstay of the treatment in newly diagnosed transplant-eligible multiple myeloma (MM) patients. This retrospective study was performed to investigate the potential prognostic markers which may modify transplant course in a total of 256 ASCT recipients [median age: 58 (30-74) years; male/female: 138/118], including pretransplant (PET0) and day + 60 (PET2) PET/CT assessments and comparative analysis of melphalan (Mel) dose. Better responses with significantly higher complete response/very good partial response rates were achieved in patients who proceeded to transplant within 301 days from diagnosis (p < 0.001). Patients who had received < 1.5 lines of treatment prior to transplant had significantly higher probability of overall survival (OS) (p = 0.004) and progression-free survival (PFS) (p < 0.001). The probability of OS was significantly higher in patients with low Eastern Cooperative Oncology Group (ECOG) performance score (PS = 0-1) (p = 0.003) and HCT-Comorbidity Index (HCT-CI = 0) (p = 0.011). The number of involved areas (p = 0.028) and maximum standardized uptake value (SUVmax) (p = 0.021) in PET0 represented significant impact on OS. The probabilities of OS (p < 0.001) and PFS (p = 0.01) were significantly better with Mel200 mg/m2 conditioning compared to Mel140 mg/m2. Conditioning with Mel200 mg/m2, early and upfront ASCT and low pretransplant treatment burden were found to be significantly associated with ASCT outcome in MM patients. Despite its predictor impact on survival and prognosis, further studies are warranted to standardize PET/CT-based response assessments before being used as a guide for treatment decisions in clinical practice.


Co-occurring SAD symptomatology and schizophrenia at high latitude: a pilot study.
Recent research investigates major depression with seasonal pattern, also called seasonal affective disorder (SAD), depression in schizophrenia and seasonality in schizophrenia, but there exits limited research investigating SAD in schizophrenia. This study documents co-occurring SAD symptomatology in patients with schizophrenia at high latitude. Clinical cross-sectional study of patients with schizophrenia attending treatment centres in Alaska. Twenty-eight patients completed a structured interview assessing seasonal patterns in mood, depression, negative symptoms of schizophrenia and alcohol use. Thirty-six percent of patients with schizophrenia met the criteria for SAD used in previous general population research on SAD in Alaska. When a presence of major depressive episode was confirmed using a structured clinical interview for depression in schizophrenia, the rate was 25%. Severity of SAD symptoms was greatest among patients with alcohol-abuse history. Co-occurring SAD symptomatology was identified in this extreme latitude sample of patients with schizophrenia. The frequency and severity of symptomatology was greater than found in a general population study of SAD conducted in Alaska using identical criteria. SAD may be under-diagnosed in schizophrenia at moderate and extreme latitudes, highlighting clinical assessment considerations, potential utility of bright light therapy and the need for additional research.


Shifting Medication Treatment Practices in the COVID-19 Pandemic: A Statewide Survey of Pennsylvania Opioid Treatment Programs.
We sought to understand how opioid treatment programs (OTPs) adapted OTP operations to the COVID-19 pandemic and new federal regulations around methadone and buprenorphine. In fall 2020, we conducted an online survey of all 103 OTPs licensed by the Pennsylvania Department of Drug and Alcohol Programs, including clinical directors. Survey domains included changes to methadone take-home and telehealth practices; overdose and diversion prevention tactics; perceptions regarding how such changes influence patient well-being; and financial/operational concerns related to the new policies and practices. We calculated descriptive statistics and conducted Chi-square test to test for differences between not-for-profit versus for-profit and large versus small OTPs. Forty-seven percent (46%) OTPs responded to the survey. 10% and 25%, respectively, endorsed offering telephone and video-based telemedicine buprenorphine induction. Sixty-six percent endorsed extending take-home supplies of methadone, but most indicated that these extensions applied to a minority of their patients. Most respondents agreed that provision of buprenorphine via telehealth and extended take-home methadone reduced patient burden in accessing medications and prevented exposure to COVID-19, while not significantly increasing risk of overdose. We did not find major differences in COVID-19 practice modifications by nonprofit status or size of OTP. In Pennsylvania, the COVID-19 pandemic led to rapid changes in provision of opioid treatment services. Findings on relatively low uptake of longer methadone take-home regimens and virtual buprenorphine initiation despite general support for these practices imply a need to further develop guidelines for best clinical practices and understand/address barriers to their implementation.


Ectopic Expression of Innate Immune Protein, Lipocalin-2, in Lactococcus lactis Protects Against Gut and Environmental Stressors.
Lipocalin-2 (Lcn2) is a multifunctional innate immune protein that exhibits antimicrobial activity by the sequestration of bacterial siderophores, regulates iron homeostasis, and augments cellular tolerance to oxidative stress. Studies in the murine model of colitis have demonstrated that Lcn2 deficiency exacerbates colitogenesis; however, the therapeutic potential of Lcn2 supplementation has yet to be elucidated. In light of its potential mucoprotective functions, we, herein, investigated whether expression of Lcn2 in the probiotic bacterium can be exploited to alleviate experimental colitis. Murine Lcn2 was cloned into the pT1NX plasmid and transformed into Lactococcus lactis to generate L. lactis-expressing Lcn2 (Lactis-Lcn2) or the empty plasmid (Lactis-Con). Lactis-Lcn2 was characterized by immunoblot and enzyme-linked immunosorbent assay and tested for its antimicrobial efficacy on Escherichia coli. The capacity of Lactis-Lcn2 and Lactis-Con to withstand adverse conditions was tested using in vitro viability assays. Dextran sodium sulfate colitis model was used to investigate the colonization ability and therapeutic potential of Lactis-Lcn2 and Lactis-Con. Lcn2 derived from Lactis-Lcn2 inhibited the growth of E. coli and reduced the bioactivity of enterobactin (E. coli-derived siderophore) in vitro. Lactis-Lcn2 displayed enhanced tolerance to adverse pH, high concentration of bile acids, and oxidative stress in vitro and survived better in the inflamed gut than Lactis-Con. Consistent with these features, Lactis-Lcn2 displayed better mucoprotection against intestinal inflammation than Lactis-Con when administered into mice with dextran sulfate sodium-induced acute colitis. Our findings suggest that Lcn2 expression can be exploited to enhance the survivability of probiotic bacteria during inflammation, which could further improve its efficacy to treat experimental colitis.


Contrast resolution of line-scanned digital radiography.
Contrast resolution of a prototype system for line-scanned digital radiography was evaluated using a phantom simulating low contrast tumors within the body. Simulated tumors having radiographic contrast as low as 0.5% were detectable in these studies. The results of these digital radiographic studies compare favorably to those of computed tomography in terms of both low contrast detectability and radiation dosage. Dual energy projection radiographic techniques show promise for eliminating obscuring shadows caused by overlying structures from digital radiographs, thus permitting clinical utilization of the excellent contrast resolution of the line scanned projection system.


Dabrafenib plus trametinib in unselected advanced BRAF V600-mut melanoma: a non-interventional, multicenter, prospective trial.
The efficacy of combined BRAF and MEK inhibition for BRAF V600-mutant melanoma in a broad patient population, including subgroups excluded from phase 3 trials, remains unanswered. This noninterventional study (DATUM-NIS) assessed the real-world efficacy, safety and tolerability of dabrafenib plus trametinib in Austrian patients with unresectable/metastatic melanoma. This multicenter, open-label, non-interventional, post-approval, observational study investigated the effectiveness of dabrafenib plus trametinib prescribed in day-to-day clinical practice to patients (N = 79) with BRAF V600-mutant unresectable/metastatic melanoma with M1c disease (American Joint Committee on Cancer staging manual version 7), ECOG > 1, and elevated serum lactate dehydrogenase (LDH). The primary endpoint was 6-, 12- and 18-month progression-free survival (PFS) rates. Secondary endpoints were median PFS, disease control rate and overall survival (OS). The 6-, 12- and 18-month PFS rates were 76%, 30.6% and 16.2%, respectively. Subgroup analysis showed a significant PFS benefit in the absence of lung metastasis. The median PFS and OS were 9.1 (95% CI, 7.1-10.3) months and 17.9 (95% CI, 12.7-27.8) months, respectively. The 12- and 24-month OS rates were 62.7% and 26.8%, respectively. Subgroup analyses showed significant OS benefits in the absence of bone or lung metastasis and the presence of other metastases (excluding bone, lung, brain, liver and lymph nodes). Furthermore, S100 and Eastern Cooperative Oncology Group performance status (ECOG PS) showed a significant impact on survival. No new safety signals were observed. Despite an unselected population of melanoma patients with higher M1c disease, ECOG PS > 1 and elevated LDH, this real-world study demonstrated comparable efficacy and safety with the pivotal phase 3 clinical trials for dabrafenib-trametinib.


Metropolitan lizards? Urbanization gradient and the density of lagartixas (Tropidurus hispidus) in a tropical city.
Urbanization, with its cohort of environmental stressors, has a dramatic effect on wildlife, causing loss of biodiversity and decline in population abundance customarily associated with increasing levels of impervious surface and fragmentation of native habitats. Some studies suggest that faunal species from open habitats, and with higher abundance in natural environments, seem more likely to tolerate and live in urban environments. Here I evaluate how the level of urbanization affects lagartixas (Tropidurus hispidus) one of the most common lizards found in open vegetation ecosystems in NE Brazil. I surveyed a total of 47 transects across sites that differed in proportion of impervious surface (high, mild, peri-urban, and rural). I also collected specific biotic (herbaceous cover, tree, and arthropod abundance) and abiotic (amount of shelters and impervious surfaces) factors that could affect lagartixas abundance. Ants were the most common arthropod both in the rural and urban environment. Lagartixas thrive in urban environments, and trees and shelter were key predictors of their abundance. Lagartixas show a clear association with use of artificial structures. The low densities of lagartixas in rural areas and higher density in urbanized areas suggest that they colonized urban areas due to the hard surfaces and they probably are not exploiting a novel habitat, but somewhat responding to conditions resembling those in which they evolved. Finally, lagartixas are extremely common in tropical cities, they have a suite of features that are associated with selective pressures in cities and they might play a key functional role in urban ecosystems making this lizard an excellent system for the study of ecology and adaptation to the urban environments.


[Angiogenesis of gynecologic tumors studied with color Doppler].
Angiogenesis is obligatory event in solid tumors growth. Weak muscular layer in the newly formed tumoral vessels results in decreased resistance to blood flow and characteristic Doppler signal. The number and arrangement of the blood vessels differ between benign and malignant lesions as well. Parameters mentioned above enable a precise ultrasound differential diagnosis between benign and malignant tumors. Malignant uterine tumors are better perfused and have lower resistance to blood flow than benign ones. Benign lesions with secondary changes (necrosis) may have lowered resistance indices. The finding of pathological perfusion makes the diagnosis of endometrial carcinoma also very accurate. High sensitivity of the transvaginal color Doppler in detecting newly formed blood vessels in an early stages of ovarian cancer may establish this technique as a screening method for this still very high lethality disease.


Development of a teletechnology protocol for in-home rehabilitation.
Our ability to provide in-home rehabilitation is limited by distance and available personnel. We may be able to meet some rehabilitation needs with videoconferencing technology. This article describes the feasibility of teletechnology for delivering multifactorial, in-home rehabilitation interventions to community-dwelling adults recently prescribed a mobility aid. We used standard telephone lines to provide two-way video and audio interaction. The interventions included prescription of and/or training in functionally based exercises, home-hazard assessment, assistive technology, environmental modifications, and adaptive strategies. Patients were evaluated in three transfer and three mobility tasks, and appropriate treatment was provided over the course of four visits. To date, 13 of the 14 subjects enrolled in the rehabilitation study have completed all four visits (56 visits total). Equipment-related problems were most common early in the study, particularly on the initial visit to a subject's house. We identified (mean +/- standard deviation [SD]) 13.1 +/- 7.9 mobility/self-care problems per subject and made 12.5 +/- 8.3 recommendations per subject to address those problems. At 6-week follow-up, 60.1 percent of our recommendations had been implemented. The greatest number of problems was identified for tub transfers (mean +/- SD = 3.4 +/- 1.4), the greatest number of recommendations was made for toilet transfers (mean +/- SD = 3.1 +/- 3.4), and the most frequently implemented recommendations were for transition between locations. Overall, our results show promise that both the telerehabilitation technology and intervention procedures are feasible.


Reversal of impaired wound repair in iNOS-deficient mice by topical adenoviral-mediated iNOS gene transfer.
Most evidence indicates that nitric oxide plays a role in normal wound repair; however, involvement of inducible nitric oxide synthase (iNOS) has not been established. Experiments were carried out to determine the requirement for iNOS in closing excisional wounds. Wound closure was delayed by 31% in iNOS knockout mice compared with wild-type animals. An identical delay in wound closure was observed in wild-type mice given a continuous infusion of the partially selective iNOS inhibitor N6-(iminoethyl)-L-lysine. Delayed wound healing in iNOS-deficient mice was completely reversed by a single application of an adenoviral vector containing human iNOS cDNA (AdiNOS) at the time of wounding. Reverse transcription PCR identified iNOS mRNA expression in wild-type mice peaking 4-6 d after wounding, and confirmed expression of human iNOS in the adenoviral vector containing human iNOS cDNA-treated animals. These results establish the key role of iNOS in wound closure, and suggest a gene therapy strategy to improve wound healing in iNOS-deficient states such as diabetes, and during steroid treatment.


Minimally Invasive Posterior Decompression Combined With Percutaneous Pedicle Screw Fixation for the Treatment of Thoracolumbar Fractures With Neurological Deficits: A Prospective Randomized Study Versus Traditional Open Posterior Surgery.
Prospective randomized cohort study. To compare the surgical results of minimally invasive posterior decompression combined with percutaneous pedicle screws fixation (minimally invasive surgery [MIS]) and posterior open surgery (OS) for the treatment of thoracolumbar fracture with neurological deficits. Thoracolumbar fracture with neurological deficits usually undergoes surgical intervention. OS can achieve satisfied results, but the main disadvantage is approach-related complications. No study, however, focused on the treatment of this disease by MIS through posterior approach. Sixty consecutive cases of thoracolumbar fractures with neurological deficits were randomized into MIS group and OS group. Incision length, blood loss, postoperative drainage volume, hospitalization days, blood transfusion rate, analgesic use rate, and x-ray exposure time were used to evaluate the perioperative information and Visual Analog Scale (VAS), Japanese Orthopedics Association (JOA) score, and American Spinal Injury Association grade for patients' symptom. For radiological assessment, sagittal Cobb angle, percentage of vertebral height, and vertebral wedging angle were measured. Fifty-nine of sixty patients were followed-up for at least 12 months. MIS group was superior in perioperative information (P < 0.05), except in the operative time (P = 0.165) and x-ray time (P = 0.000). The operative time seemed longer in MIS group, but no significant difference was found. The x-ray time was significantly higher in MIS group. The mean Visual Analog Scale and Japanese Orthopedics Association scores of the final follow-up in MIS group were better than that in OS group (P < 0.05). Patients in both group achieved a similar neurological recovery according to American Spinal Injury Association grade (P = 0.760). A broken screw was found in one patient in MIS group and a broken rod in one patient in OS group. MIS group has achieved the similar effect of OS group and it can minimize the approach-related complication. It also faced with some shortages, such as larger radiation dose and longer learning curve. 2.


Prevalence of mental health conditions and pain in adults with skeletal dysplasia.
We sought to examine the prevalence of depression and anxiety in adults with skeletal dysplasias, and to assess any correlations with pain. Participation was via an anonymous REDCap survey, which consisted of sociodemographic questions followed by the brief pain inventory-short form (BPI-SF), patient health questionnaire-8 (PHQ-8), and generalized anxiety disorder-7 (GAD-7) questionnaires. These assessed pain, depression, and anxiety respectively. Of the 336 usable responses, 16.1% scored 10 or greater on the PHQ-8 consistent with current depression while 17.3% scored 10 or greater on the GAD-7 consistent with current anxiety. The majority of participants (76.2%) experienced pain, which was significantly associated with prior mental health diagnoses (p < 0.05). A total of 34% reported either a prior diagnosis of depression or scored 10 or greater on the PHQ-8, and 31% reported either a prior diagnosis of anxiety or scored 10 or greater on the GAD-7. This study identified a substantial percentage of individuals with mental health concerns as well as pain in the adult skeletal dysplasia population. Further research is warranted to investigate barriers to service or treatment of mental health disorders as well as pain management.


Severe electrolyte disorders following cardiac surgery: a prospective controlled observational study.
Electrolyte disorders are an important cause of ventricular and supraventricular arrhythmias as well as various other complications in the intensive care unit. Patients undergoing cardiac surgery are at risk for development of tachyarrhythmias, especially in the period during and immediately after surgical intervention. Preventing electrolyte disorders is thus an important goal of therapy in such patients. However, although levels of potassium are usually measured regularly in these patients, other electrolytes such as magnesium, phosphate and calcium are measured far less frequently. We hypothesized that patients undergoing cardiac surgical procedures might be at risk for electrolyte depletion, and we therefore conducted the present study to assess electrolyte levels in such patients. Levels of magnesium, phosphate, potassium, calcium and sodium were measured in 500 consecutive patients undergoing various cardiac surgical procedures who required extracorporeal circulation (group 1). A total of 250 patients admitted to the intensive care unit following other major surgical procedures served as control individuals (group 2). Urine electrolyte excretion was measured in a subgroup of 50 patients in both groups. All cardiac patients received 1 l cardioplegia solution containing 16 mmol potassium and 16 mmol magnesium. In addition, intravenous potassium supplementation was greater in cardiac surgery patients (mean +/- standard error: 10.2 +/- 4.8 mmol/hour in cardiac surgery patients versus 1.3 +/- 1.0 in control individuals; P < 0.01), and most (76% versus 2%; P < 0.01) received one or more doses of magnesium (on average 2.1 g) for clinical reasons, mostly intraoperative arrhythmia. Despite these differences in supplementation, electrolyte levels decreased significantly in cardiac surgery patients, most of whom (88% of cardiac surgery patients versus 20% of control individuals; P < 0.001) met criteria for clinical deficiency in one or more electrolytes. Electrolyte levels were as follows (mmol/l [mean +/- standard error]; cardiac patients versus control individuals): phosphate 0.43 +/- 0.22 versus 0.92 +/- 0.32 (P < 0.001); magnesium 0.62 +/- 0.24 versus 0.95 +/- 0.27 (P < 0.001); calcium 1.96 +/- 0.41 versus 2.12 +/- 0.33 (P < 0.001); and potassium 3.6 +/- 0.70 versus 3.9 +/- 0.63 (P < 0.01). Magnesium levels in patients who had not received supplementation were 0.47 +/- 0.16 mmol/l in group 1 and 0.95 +/- 0.26 mmol/l in group 2 (P < 0.001). Urinary excretion of potassium, magnesium and phosphate was high in group 1 (data not shown), but this alone could not completely account for the observed electrolyte depletion. Patients undergoing cardiac surgery with extracorporeal circulation are at high risk for electrolyte depletion, despite supplementation of some electrolytes, such as potassium. The probable mechanism is a combination of increased urinary excretion and intracellular shift induced by a combination of extracorporeal circulation and decreased body temperature during surgery (hypothermia induced diuresis). Our findings may partly explain the high risk of tachyarrhythmia in patients who have undergone cardiac surgery. Prophylactic supplementation of potassium, magnesium and phosphate should be seriously considered in all patients undergoing cardiac surgical procedures, both during surgery and in the immediate postoperative period. Levels of these electrolytes should be monitored frequently in such patients.


Elevation of serum gonadotropins establishes the diagnosis of anorchism in prepubertal boys with bilateral cryptorchidism.
Of more than 500 boys with bilateral cryptorchidism who presented during a 10-year period 28 prepubertal patients less than 11 years old who had no palpable testes after human chorionic gonadotropin therapy were studied to assess the ability of serum gonadotropin levels to identify patients with anorchism. Of the boys 21 had a normal testosterone response to human chorionic gonadotropin therapy and all of them had testes at exploration. The serum luteinizing hormone levels ranged from 2 to 6 mIU per ml., with a mean of 3.7 mIU per ml., and the serum follicle-stimulating hormone levels ranged from 1.6 to 6.2 mIU per ml., with a mean of 3.7 mIU per ml. Seven patients showed no testosterone response to human chorionic gonadotropin and all but 1 underwent exploration, at which time no testes were found. Of these 7 patients 6 had elevated gonadotropin levels that averaged 3 standard deviations above the mean. For comparison, 2 pubertal patients with nonpalpable gonads and 3 castrated prepubertal boys also were studied. From the study we concluded that in boys with nonpalpable gonads 1) abnormally elevated serum gonadotropin levels before puberty are indicative of anorchism, 2) neither exploration nor human chorionic gonadotropin stimulation tests are essential for diagnosis in these select patients, 3) serum gonadotropin levels alone are not sufficient for a definitive diagnosis after puberty and 4) all boys with normal serum gonadotropin levels must undergo exploration regardless of the outcome of a human chorionic gonadotropin stimulation test.


Effects of dietary fat on the growth of normal, preneoplastic and neoplastic mammary epithelial cells in vivo and in vitro.
In order to determine: (1) whether there is a growth-regulating interaction between the mammary fat pad and mammary epithelium; (2) whether this interaction could be modified by dietary fats; and (3) whether these effects could be demonstrated in vitro, the following experiments were performed. Virgin Balb/c mice had the left inguinal mammary fat pad cleared of epithelium and were then maintained on one of four fully defined diets. These diets contained the following proportions of fat by weight: 5% or 10% mixed fats; 20% saturated fat plus cholesterol; or 20% polyunsaturated fat. To test for effects in vivo, animals received subcutaneous injections into the cleared fat pad of tumorigenic mammary cells (WAZ-2T(+SA) or WAZ-2T(-SA)) or preneoplastic mammary cells (CL-S1). Dietary fat had little effect on the latent period of tumour formation, but a low-fat diet increased the invasive/metastatic potential of both tumorigenic cell lines. A high-saturated-fat diet inhibited the growth of normal and preneoplastic epithelium in vivo. To test for effects in vitro, CL-S1 cells were co-cultured with explants of cleared mammary fat pad embedded within collagen gels. CL-S1 cells co-cultured with adipose explants obtained from mice fed on a diet containing 20% polyunsaturated fat showed a threefold increase in incorporation of [3H]thymidine into trichloroacetic acid-precipitable material. These results imply that dietary fats may affect the growth of mammary epithelium in two ways: the inhibition of growth caused by the high-saturated-fat diet may be due to systemic effects as it was not apparent in vitro; the increase in growth seen in vitro and caused by a high-polyunsaturated-fat diet is due to a direct interaction between the mammary fat pad and mammary epithelial cells. This interaction may be masked by systemic effects in vivo.


Mental health workers' views on addressing tobacco use.
The aims of this study were to assess mental health workers' attitudes to addressing tobacco dependence with patients, to identify barriers that may prevent people with mental illness from receiving support, and determine workers' recommendations for tobacco policy and practice change. Questionnaires were sent to government and non-government mental health services in Adelaide, South Australia. The questionnaires asked workers to rate on a Likert rating scale their attitudes to addressing tobacco use with patients and to respond to qualitative questions about managing tobacco use within mental health services. Quantitative data were analysed using SPSS version 15.0 and qualitative data was coded by three researchers into categories or themes and then analysed. Questionnaires (324) were received from staff across 45 organizations providing clinical and psychosocial services. More than two thirds of the participants felt it was important for mental health services to be involved in assisting patients to quit smoking; however, only 26% said they raised the issue of tobacco use with patients, often or as part of the assessment. Respondents on average felt approximately one third of their patients could quit or reduce tobacco use if they received nicotine replacement therapy, information and support. Most workers suggested changes to policy and practice within mental health services including smoke free areas and the provision of information and support. The rate of tobacco use amongst people with mental illness is still very high with serious social and health consequences. Mental health workers are uniquely placed to assist patients to quit or reduce tobacco use. This study has shown many workers believe that it is important to address tobacco use with their patients as part of routine care and that mental health services should implement significant tobacco policy and practice change.


Psychotropic Medication Effects on Seizure Threshold and Seizure Duration During Electroconvulsive Therapy Stimulus Titration.
Decisions about psychotropic medication administration before electroconvulsive therapy (ECT) are central to management of a very psychiatrically ill patient population. Given that many psychotropic medications are thought to either promote or prevent seizures, there is ongoing concern about concurrent psychotropic medication and ECT administration. This study examined the effect of psychotropic medications on seizure threshold and duration during ECT stimulus titration. The study sample consisted of 550 patients receiving ECT stimulus titration at a single site during a 27-month period. Systematic chart review provided clinical data, including patients' demographics, psychiatric diagnoses, medications administered in the 48 hours before ECT, and information on the ECT procedure. Referring psychiatrists were advised to discontinue lithium before ECT but otherwise managed psychotropic medications as clinically indicated. A fixed charge titration schedule was used to estimate seizure threshold. Electroconvulsive therapy motor seizure duration was estimated by the cuff method, and electroencephalogram seizure duration was estimated by review of a 2-lead strip. Administration of psychotropic medications, including benzodiazepines, antiepileptics, selective serotonin reuptake inhibitors, tricyclic and tetracyclic antidepressants, bupropion, and stimulants, was not associated with seizure threshold as estimated by electrical charge eliciting a generalized seizure or duration during the initial ECT titration. Tricyclic and tetracyclic antidepressant dosage was associated with seizure threshold. Psychotropic medications may have little effect on seizure threshold and duration during titration of electrical dose at ECT initiation. Integrating this work with other literature supports making recommendations for medication discontinuation before ECT on an individual basis.


Diversity of begomoviruses associated with mosaic disease of cultivated cassava (Manihot esculenta Crantz) and its wild relative (Manihot glaziovii Mull. Arg.) in Uganda.
Cassava (Manihot esculenta) growing in Uganda during 2001-2002 has been screened for the presence of begomoviruses using PCR-RFLP, cloning full-length genomic components and nucleotide sequence analysis. In contrast with a recent survey in neighbouring Kenya, which identified three distinct strains of East African cassava mosaic virus (EACMV, EACMV-UG and EACMV-KE2) as well as East African cassava mosaic Zanzibar virus and the new species East African cassava mosaic Kenya virus, only EACMV-UG and, to a lesser extent, African cassava mosaic virus (ACMV) were found associated with cassava in Uganda. The integrity of the cloned genomic components of representative virus isolates was confirmed by demonstrating their infectivity in Nicotiana benthamiana and cassava using biolistic inoculation, providing a convenient means to screen cassava varieties for disease resistance. Both EACMV-UG and ACMV were also associated with Manihot glaziovii. Infectivity studies using cloned components confirmed that viruses from one host could infect the other, suggesting that this wild relative of cassava might be a reservoir host for the disease. The relatively low level of diversity of begomoviruses associated with cassava mosaic disease in Uganda is consistent with reports that EACMV-UG has displaced other begomovirus species and strains during the recent epidemic that swept through the country.


Serotonin transporter polymorphisms (SLC6A4 insertion/deletion and rs25531) do not affect the availability of 5-HTT to [11C] DASB binding in the living human brain.
Studies in vitro suggest that the expression of the serotonin transporter (5-HTT) is regulated by polymorphic variation in the promoter region of the 5-HTT gene (5-HTTLPR); however, results from human brain imaging studies examining the relation between 5-HTT genotype and 5-HTT radioligand binding in vivo have been inconsistent. This inconsistency could reflect small participant numbers or the use of sub-optimal radiotracer for measuring the 5-HTT. We used positron emission tomography in conjunction with the selective 5-HTT ligand [(11)C] DASB to examine the availability of the 5-HTT in seven brain regions in 63 healthy European caucasian volunteers who were genotyped for short (S) and long (L) variants (SLC6A4 and rs25531) of the 5-HTTLPR. [(11)C] DASB binding potential was not influenced by the allelic status of participants whether classified on a biallelic or triallelic basis in any of the regions studied. Our PET findings, in a relatively large sample with a near optimal radiotracer, suggest that 5-HTTLPR polymorphic variation does not affect the availability of 5-HTT to [(11)C] DASB binding in adult human brain. The reported impact of 5-HTTLPR polymorphic variation on emotional processing and vulnerability to depression are more likely therefore to be expressed through effects exerted during neurodevelopment.


Flow dependence of chloride transport in rat S1 proximal tubules.
These studies examined whether the luminal flow dependency of chloride absorption in the S1 proximal tubule during glomerulotubular balance was due to change in active and/or passive transport of chloride. Using in vivo microperfusion in the Munich-Wistar rat and an essentially pure sodium chloride perfusate (devoid of bicarbonate and organic solutes), we found that an increase in luminal perfusion rate from 30 to 45 nl/min caused stimulation of total chloride absorption (active plus passive) by 87 peq.mm-1.min-1 (632 +/- 17 to 719 +/- 11, P < 0.001). When cyanide was added to this perfusate to eliminate active transport, the flow-induced change in passive transport was 58 peq.mm-1.min-1 (479 +/- 9 to 537 +/- 11, P < 0.001). The cyanide-inhibitable active transport component was therefore 29 peq.mm-1.min-1. With elimination of the transepithelial chloride gradient and, hence, passive transport by isethionate substitution, active transport increased by 63 peq.mm-1.min-1 (121 +/- 4 to 184 +/- 7, P < 0.001) as flow rate rose from 30 to 45nl/min. Removal of organic solutes from a glomerular ultrafiltrate-like perfusate had a minimal effect on flow-induced change in chloride transport (190 vs. 207 peq.mm-1.min-1). In conclusion, flow-dependent active and passive chloride transport in the S1 proximal tubule may both contribute to normal glomerulotubular balance.


Center of pressure measures to assess standing performance.
Functional neuromuscular stimulation (FNS) systems have provided individuals with spinal cord injury (SCI) varying levels of standing capability. Our long-term goal is to enhance an FNS user's ability to perform activities of daily living by providing sufficient standing stability. However, control of standing posture with complete SCI is compromised by a lack of sensory feedback from receptors in the feet. The goals of this study were: a) to document a variety of center of pressure (COP) measures during standing, b) to compare able-bodied and FNS standing, and c) to relate various measures to overall quality of standing performance. Two subjects with complete SCI that stand with FNS (one with a surface system and one with an implanted stimulation system) and five able-bodied subjects stood quietly on force platforms. Reduced hand support forces were coincident with anterior/posterior (A/P) mean COPs that were further posterior in the base of support and medial/lateral (M/L) mean COPs that were more centered between the feet. In addition, M/L COP excursions, maximum A/P COP velocities, and maximum M/L COP velocities were all significantly greater for FNS users than for able-bodied standers. Furthermore, maximum A/P COP velocities and mean A/P COP frequencies were significantly greater during FNS standing trails that required increased support from the hands. COP measures such as these may be useful tools to assess standing performance and to determine strategies for adjusting posture that can reduce an FNS user's reliance on upper extremity support while maintaining stable balance.


Brownian motion as a new probe of wettability.
Understanding wettability is crucial for optimizing oil recovery, semiconductor manufacturing, pharmaceutical industry, and electrowetting. In this letter, we study the effects of wettability on Brownian motion. We consider the cases of a sphere in an unbounded fluid medium, as well as a sphere placed in the vicinity of a plane wall. For the first case, we show the effects of wettability on the statistical properties of the particles' motion, such as velocity autocorrelation, velocity, and thermal force power spectra over a large range of time scales. We also propose a new method to measure wettability based on the particles' Brownian motion. In addition, we compare the boundary effects on Brownian motion imposed by both no-slip and perfect-slip flat walls. We emphasize the surprising boundary effects on Brownian motion imposed by a perfect-slip wall in the parallel direction, such as a higher particle mobility parallel to a perfect flat wall compared to that in the absence of the wall, as well as compared to a particle near a no-slip flat wall.


Rapid purification and molecular modeling of AaIT peptides from venom of Androctonus australis.
As recombinant viruses expressing scorpion toxins are moving closer toward the market, it is important to obtain large amounts of pure toxin for biochemical characterization and the evaluation of biological activity in nontarget organisms. In the past, we purified a large amount of Androctonus australis anti-insect toxin (AaIT) present in the venom of A. australis with an analytical reversed-phase column by repeated runs of crude sample. We now report 20 times improved efficiency and speed of the purification by employing a preparative reversed-phase column. In just two consecutive HPLC steps, almost 1 mg of AaIT was obtained from 70 mg crude venom. Furthermore, additional AaIT was obtained from side fractions in a second HPLC run. Recently discovered insect selective toxin, AaIT5, was isolated simultaneously from the same venom batch. It shows different biological toxicity symptoms than the known excitatory and depressant insect toxins. AaIT5 gave 100% mortality with a dose of less than 1.3 micrograms against fourth-instar tobacco budworms Heliothis virescens 24 h after injection. During the purification process, we implemented mass spectrometry in addition to bioassays to monitor the presence of AaIT and AaIT5 in the HPLC fractions. Mass spectrometric screening can unambiguously follow the purification process and can greatly facilitate and expedite the downstream purification of AaIT and AaIT5 eliminating the number of bioassays required. Further, electrospray ionization was compared with matrix-assisted desorption/ionization and evaluated as a method of choice for mass spectrometric characterization of fractions from the venom purification for it provided higher mass accuracy and relative quantitation capability. Molecular models were built for AaIT5, excitatory toxin AaIT4, and depressant toxin LqhIT2. Three-dimensional structure of AaIT5 was compared with structures of the other two toxins, suggesting that AaIT5 is similar to depressant toxins.


Extrapolation of Metal Toxicity Data for the Rotifer Brachionus calyciflorus Using an Individual-Based Population Model.
Ecological risk assessment (ERA) of metals typically starts from standardized toxicity tests, the data from which are then extrapolated to derive safe concentrations for the envisioned protection goals. Because such extrapolation in conventional ERA lacks ecological realism, ecological modeling is considered as a promising new approach for extrapolation. Many published population models are complex, that is, they include many processes and parameters, and thus require an extensive dataset to calibrate. In the present study, we investigated how individual-based models based on a reduced version of the Dynamic Energy Budget theory (DEBkiss IBM) could be applied for metal effects on the rotifer Brachionus calyciflorus. Data on survival over time and reproduction at different temperatures and food conditions were used to calibrate and evaluate the model for copper effects. While population growth and decline were well predicted, the underprediction of population density and the mismatch in the onset of copper effects were attributed to the simplicity of the approach. The DEBkiss IBM was applied to toxicity datasets for copper, nickel, and zinc. Predicted effect concentrations for these metals based on the maximum population growth rate were between 0.7 and 3 times higher in all but one case (10 times higher) than effect concentrations based on the toxicity data. The size of the difference depended on certain characteristics of the toxicity data: both the steepness of the concentration-effect curve and the relative sensitivity of lethal and sublethal effects played a role. Overall, the present study is an example of how a population model with reduced complexity can be useful for metal ERA. Environ Toxicol Chem 2023;00:1-14. © 2023 SETAC.


Semiring chemistry of Au25(SR)18: fragmentation pathway and catalytic active site.
The semiring chemistry of the Au25(SR)18, particularly its fragmentation mechanism and catalytic active site, is explored using density functional theory (DFT) calculations. Our calculations show that the magically stable fragmental cluster, Au21(SR)14(-), as detected in several mass spectrometry (MS) measurements of fragmentation of the Au25(SR)18(-), contains a quasi-icosahedral Au13-core fully protected by four -SR-Au-SR- and two -SR-Au-SR-Au-SR- staple motifs. A stepwise fragmentation mechanism of the semiring staple motifs on the surface of Au25(SR)18(-) is proposed for the first time. Initially, the Au25(SR)18(-) transforms into a metastable structure with all staple motifs binding with two neighboring vertex Au-atoms of the Au-core upon energy uptake. Subsequently, a 'step-by-step' detachment and transfer of [Au(SR)]x (x = 1-4) units occurs, which leads to the formation of highly stable products including Au21(SR)14(-) and a cyclic [Au(SR)]4 unit. The continued fragmentation of Au21(SR)14(-) to Au17(SR)10(-) is observed as well, which shows same stepwise fragmentation mechanism. The proposed mechanism well explains the favorable formation of Au21(SR)14(-) and Au17(SR)10(-) from Au25(SR)18(-) as observed from experimental abundance. Taking the Au21(SR)14 and its parent cluster Au25(SR)18 as the benchmark model systems, the catalytic active site of the thiolate protected gold clusters toward the styrene oxidation and the associated reaction mechanism are further investigated. We show that the Au atom in the staple motifs is the major active site for the styrene oxidation in presence of TBHP as oxidant or initiator. The Au atom in the staple motifs can change from Au(I) (bicoordinated) to Au(III) (tetracoordinated). The O2 activation is achieved during this process.


Extracellular vesicles from the apoplastic fungal wheat pathogen Zymoseptoria tritici.
The fungal pathogen Zymoseptoria tritici is a significant constraint to wheat production in temperate cropping regions around the world. Despite its agronomic impacts, the mechanisms allowing the pathogen to asymptomatically invade and grow in the apoplast of wheat leaves before causing extensive host cell death remain elusive. Given recent evidence of extracellular vesicles (EVs)-secreted, membrane-bound nanoparticles containing molecular cargo-being implicated in extracellular communication between plants and fungal pathogen, we have initiated an in vitro investigation of EVs from this apoplastic fungal wheat pathogen. We aimed to isolate EVs from Z. tritici broth cultures and examine their protein composition in relation to the soluble protein in the culture filtrate and to existing fungal EV proteomes. Zymoseptoria tritici EVs were isolated from broth culture filtrates using differential ultracentrifugation (DUC) and examined with transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA). Z. tritici EVs were observed as a heterogeneous population of particles, with most between 50 and 250 nm. These particles were found in abundance in the culture filtrates of viable Z. tritici cultures, but not heat-killed cultures incubated for an equivalent time and of comparable biomass. Bottom-up proteomic analysis using LC-MS/MS, followed by stringent filtering revealed 240 Z. tritici EV proteins. These proteins were distinct from soluble proteins identified in Z. tritici culture filtrates, but were similar to proteins identified in EVs from other fungi, based on sequence similarity analyses. Notably, a putative marker protein recently identified in Candida albicans EVs was also consistently detected in Z. tritici EVs. We have shown EVs can be isolated from the devastating fungal wheat pathogen Z. tritici and are similar to protein composition to previously characterised fungal EVs. EVs from human pathogenic fungi are implicated in virulence, but the role of EVs in the interaction of phytopathogenic fungi and their hosts is unknown. These in vitro analyses provide a basis for expanding investigations of Z. tritici EVs in planta, to examine their involvement in the infection process of this apoplastic wheat pathogen and more broadly, advance understanding of noncanonical secretion in filamentous plant pathogens.