Surgical intensive care - current and future challenges?
Bjorn Ibsen, an anesthetist who pioneered positive pressure ventilation as a treatment option during the Copenhagen polio epidemic of 1952, set up the first Intensive Care Unit (ICU) in Europe in 1953. He managed polio patients on positive pressure ventilation together with physicians and physiologists in a dedicated ward, where one nurse was assigned to each patient. In that sense Ibsen is more or less the father of intensive care medicine as a specialty and also an advocate of the one-to-one nursing ratio for critically ill patients. Nowadays, the Surgical Intensive Care Unit (SICU) offers critical care treatment to unstable, severely, or potentially severely ill patients in the perioperative setting, who have life-threatening conditions and require comprehensive care, constant monitoring, and possible emergency interventions. Hence there is one very specific challenge in the surgical setting: the intensivist has to manage the patient flow starting from admission to the hospital through to the operating theater, in the SICU, and postoperatively for the discharge to the ward. In other words, the planning of the resources (most frequently availability of beds) has to be optimized to prevent cancellations of elective surgical procedures but also to facilitate other emergency admissions. SICU intensivists take the role of arbitrators between surgical demand and patient's interests. This means they supervise the safety, efficacy, and workability of the process with respect to all stakeholders. This notion was reported in 2007 when Stawicki and co-workers performed a small prospective study concluding that it appears safe if the dedicated intensivist takes over the role of the last arbitrator supported by a multidisciplinary team.1 However, demographic changes in many countries during the last few decades have given rise to populations which are more elderly and sicker than before. This impacts on the healthcare system in general but on the intensivist and the ICU team too. In addition, in a society with an increased life expectancy, the balance between treatable disease, outcome, and utilization of resources must be maintained. This fact gains even more importance as patients and their families claim "high end" treatment. Such a demand is reflected looking at the developments that have taken place over the last 25 years. Mainly, the focus of intensive care medicine was on technical support or even replacement of failing organ systems such as the lungs, the heart, or the kidneys by veno-venous extracorporeal membrane oxygenation (VV-ECMO), veno-arterial ECMO (VA-ECMO), and continuous veno-venous hemofiltration (CVVH) respectively. This means "technical care" became a core capability and expectation of critical care medicine. In parallel, medical treatment became more standardized. For example, lung protective ventilation strategies, early enteral feeding, and daily sedation vacation are part of modern protocols. As a consequence, ventilator time has been reduced and patients therefore develop delirium less frequently. These measures, beside others, are implemented in care bundles to improve the quality of care of patients by the whole ICU team. The importance of specialty trained teams was already pointed out 35 years ago when Li et al.,2 demonstrated in a study performed in a community hospital that the mortality was decreased if an ICU was managed 24/7 by an on-site physician. The association of improved outcomes and presence of a critical care trained physician (intensivist) has been shown in several studies since that time.3,4,5,6 A modern multidisciplinary critical care team consists at least of an intensivist, ICU nurse, pharmacist, respiratory therapist, physiotherapist, and the primary team physician. Based on clinical needs, the team can be supplemented by oncologists, cardiologists, or other specialties. Again, this approach is supported by research: a recent retrospective cohort study from the California Hospital Assessment and Reporting Taskforce (CHART) on 60,330 patients confirmed the association between improved patient outcome and such a multidisciplinary team.7 If such an intensive care team makes a difference, why do not all patients at risk receive advanced ICU-care? It was already demonstrated by Esteban et al., in a prospective study that patients with severe sepsis had a mortality rate of 26% when not admitted to an ICU in comparison to 11% when they were admitted to an ICU.8 Meanwhile, we know that early referral is particularly important, because for ischemic diseases the timing appears to make a difference in terms of full recovery. So, the following questions arise: Should intensive care be rolled out to each ward and physical admission to an ICU or be restricted to special cases only? For this purpose, the so-called "Rapid Response Teams" (RRT) or "Medical Emergency Team" (MET), which essentially are a form of an ICU outreach team, were implemented. The name, composition, or exact role of such team varies from institution to institution and country to country. Alternatively, should all ward staff be educated to recognize sick patients earlier for a timely transfer to a dedicated area? This would mean that ICU-care would be introduced in the ward. A first attempt to answer this question, whether to deploy critical care resources to deteriorating patients outside the ICU 24/7, was given by Churpek et al.9 The success of the rapid response teams could be related to decreased rates of cardiac arrest outside the ICU setting and in-hospital mortality. Interestingly, an analysis of the registry database of the RRT calls in this study showed that the lowest frequency of calls occurred between 1:00 AM to 6:59 AM time period. In contrast, the mortality was highest around 7 AM and lowest during noon hour. This indicates that not simply the availability of such a team makes a difference but also the alertness of the ward-teams is of high importance to identify deteriorating patients in a timely manner. Essentially, this would necessitate ward staff being trained to provide a higher level of care enabling them to better recognize when patients become sicker to avoid a delayed call to the ICU. Alternatively, a system in which the intensivist plays a major role in daily ward rounds could be beneficial. So, the ward doctor should become an intensivist. However, the latter means the ICU is rolled out across the whole hospital which would consume a huge amount of resources. Another option would be 24/7 remote monitoring of patients at risk that notifies the intensivist or RRT in case of need. The infrastructure, technology, and manpower to put this in place also has associated costs. As the demand for ICU care will rise further in the future, intensivists will play an even more important role in the healthcare system that itself is under enormous economic pressure to ensure the best quality of care for critically ill patients. Besides excellent knowledge and hard skills, intensivists need to be team players, communicators, facilitators, and arbitrators to achieve the best results in collaboration with all involved in patient treatment.


MISFITS: evaluating the goodness of fit between a phylogenetic model and an alignment.
As models of sequence evolution become more and more complicated, many criteria for model selection have been proposed, and tools are available to select the best model for an alignment under a particular criterion. However, in many instances the selected model fails to explain the data adequately as reflected by large deviations between observed pattern frequencies and the corresponding expectation. We present MISFITS, an approach to evaluate the goodness of fit (http://www.cibiv.at/software/misfits). MISFITS introduces a minimum number of "extra substitutions" on the inferred tree to provide a biologically motivated explanation why the alignment may deviate from expectation. These extra substitutions plus the evolutionary model then fully explain the alignment. We illustrate the method on several examples and then give a survey about the goodness of fit of the selected models to the alignments in the PANDIT database.


Magnetic resonance imaging anesthesia: new challenges and techniques.
The increasing use of magnetic resonance imaging as a diagnostic modality has led to increased demand for sedation and monitoring during the procedure. This review is to acquaint the reader with the most recent developments in magnetic resonance imaging diagnostics and to describe the evolving techniques and strategies for patient management. Many centers are meeting the challenges of increasing demand by streamlining their sedation/anesthetic protocols to achieve greater efficiency. Some have enlisted the help of nursing staff who are trained to provide sedation for certain patients. Continued experience in magnetic resonance imaging anesthesia has led to a better understanding of patient needs and decreased the number of failed procedures. The scope of magnetic resonance imaging diagnostics has expanded to include urology, otolaryngology, and neonatal evaluation. Although infants and children constitute the majority of patients, many adults also require anesthesia for magnetic resonance imaging and present their own challenges. Anesthesia and sedation during magnetic resonance imaging have a unique set of constraints. However, most of the standards of modern, safe anesthetic care can be met in this environment. The growing experience at many hospitals has demonstrated that a wide range of patients can receive safe care during magnetic resonance imaging.


Type A behaviour and pressor response in a representative sample of middle-aged men.
The hypothesis that type A behaviour is associated with greater pressor response to stress was tested in a representative sample of 114 men aged 30-65 yr. Jenkins Activity Survey, Framingham and Bortner type A scores were related to blood pressure and heart rate under rest, serial reaction time, mental arithmetic, and noise conditions. Type A scores were modestly intercorrelated (r = 0.59 to r = 0.67). Type A scores were inversely related to age and resting systolic pressure and were independent of smoking history, social class, marital status, and task performance parameters. Type A scores were not related to pressor or heart rate response to stress under any of the stress conditions. These findings do not support the importance of physiological response as an explanation of the association between type A and heart disease.


Challenges of interprofessional team training: a qualitative analysis of residents' perceptions.
Simulation-based interprofessional team training is thought to improve patient care. Participating teams often consist of both experienced providers and trainees, which likely impacts team dynamics, particularly when a resident leads the team. Although similar team composition is found in real-life, debriefing after simulations puts a spotlight on team interactions and in particular on residents in the role of team leader. The goal of the current study was to explore residents' perceptions of simulation-based interprofessional team training. This was a secondary analysis of a study of residents in the pediatric residency training program at the University of California, San Francisco (United States) leading interprofessional teams in simulated resuscitations, followed by facilitated debriefing. Residents participated in individual, semi-structured, audio-recorded interviews within one month of the simulation. The original study aimed to examine residents' self-assessment of leadership skills, and during analysis we encountered numerous comments regarding the interprofessional nature of the simulation training. We therefore performed a secondary analysis of the interview transcripts. We followed an iterative process to create a coding scheme, and used interprofessional learning and practice as sensitizing concepts to extract relevant themes. 16 residents participated in the study. Residents felt that simulated resuscitations were helpful but anxiety provoking, largely due to interprofessional dynamics. They embraced the interprofessional training opportunity and appreciated hearing other healthcare providers' perspectives, but questioned the value of interprofessional debriefing. They identified the need to maintain positive relationships with colleagues in light of the teams' complex hierarchy as a barrier to candid feedback. Pediatric residents in our study appreciated the opportunity to participate in interprofessional team training but were conflicted about the value of feedback and debriefing in this setting. These data indicate that the optimal approach to such interprofessional education activities deserves further study.


New technologies - new insights into the pathogenesis of hepatic encephalopathy.
Hepatic encephalopathy (HE) is a neuropsychiatric syndrome which frequently accompanies acute or chronic liver disease. It is characterized by a variety of symptoms of different severity such as cognitive deficits and impaired motor functions. Currently, HE is seen as a consequence of a low grade cerebral oedema associated with the formation of cerebral oxidative stress and deranged cerebral oscillatory networks. However, the pathogenesis of HE is still incompletely understood as liver dysfunction triggers exceptionally complex metabolic derangements in the body which need to be investigated by appropriate technologies. This review summarizes technological approaches presented at the ISHEN conference 2014 in London which may help to gain new insights into the pathogenesis of HE. Dynamic in vivo 13C nuclear magnetic resonance spectroscopy was performed to analyse effects of chronic liver failure in rats on brain energy metabolism. By using a genomics approach, microRNA expression changes were identified in plasma of animals with acute liver failure which may be involved in interorgan interactions and which may serve as organ-specific biomarkers for tissue damage during acute liver failure. Genomics were also applied to analyse glutaminase gene polymorphisms in patients with liver cirrhosis indicating that haplotype-dependent glutaminase activity is an important pathogenic factor in HE. Metabonomics represents a promising approach to better understand HE, by capturing the systems level metabolic changes associated with disease in individuals, and enabling monitoring of metabolic phenotypes in real time, over a time course and in response to treatment, to better inform clinical decision making. Targeted fluxomics allow the determination of metabolic reaction rates thereby discriminating metabolite level changes in HE in terms of production, consumption and clearance.


Most Marketplace Plans Included At Least 25 Percent Of Local-Area Physicians, But Enrollment Disparities Remained.
The Affordable Care Act allows commercial insurers participating in the Marketplaces to vary the size of their provider networks as long as the providers are "sufficient" in numbers and types. Concerns have been growing over the increasing use of restricted-provider or narrow networks in Marketplace plans because of their implications for reduced access to care, but little is known about the breadth and stability of these networks over time or what types of enrollees choose such plans. Using national data, we found that in 2016, 60 percent of provider networks in plans offered in the federally facilitated Marketplaces included at least one-quarter of local-area physicians, and that consumers' access to broad-network plans remained stable between 2015 and 2016. Hispanic and low-income people made up a disproportionate share of enrollees in smaller-network plans (those with fewer than one-quarter of local-area physicians). It will be important to monitor the impact of narrow networks on access to and quality of care as well as on health outcomes.


Aortic Curvature Is a Predictor of Late Type Ia Endoleak and Migration After Endovascular Aneurysm Repair.
To evaluate the association between aortic curvature and other preoperative anatomical characteristics and late (>1 year) type Ia endoleak and endograft migration in endovascular aneurysm repair (EVAR) patients. Eight high-volume EVAR centers contributed 116 EVAR patients (mean age 81±7 years; 103 men) to the study: 36 patients (mean age 82±7 years; 31 men) with endograft migration and/or type Ia endoleak diagnosed >1 year after the initial EVAR and 80 controls without early or late complications. Aortic curvature was calculated from the preoperative computed tomography scan as the maximum and average curvature over 5 predefined aortic segments: the entire infrarenal aortic neck, aneurysm sac, and the suprarenal, juxtarenal, and infrarenal aorta. Other morphological characteristics included neck length, neck diameter, mural neck calcification and thrombus, suprarenal and infrarenal angulation, and largest aneurysm sac diameter. Independent risk factors were identified using backward stepwise logistic regression. Relevant cutoff values for each of the variables in the final regression model were determined with the receiver operator characteristic curve. Logistic regression identified maximum curvature over the length of the aneurysm sac (>47 m-1; p=0.023), largest aneurysm sac diameter (>56 mm; p=0.028), and mural neck thrombus (>11° circumference; p<0.001) as independent predictors of late migration and type Ia endoleak. Aortic curvature is a predictor for late type Ia endoleak and endograft migration after EVAR. These findings suggest that aortic curvature is a better parameter than angulation to predict post-EVAR failure and should be included as a hostile neck parameter.


Col1A1 production and apoptotic resistance in TGF-β1-induced epithelial-to-mesenchymal transition-like phenotype of 603B cells.
Recent studies have suggested that proliferating cholangiocytes have an important role in the induction of fibrosis, either directly via epithelial-to-mesenchymal transition (EMT), or indirectly via activation of other liver cell types. Transforming growth factor beta 1 (TGF-β1), a critical fibrotic cytokine for hepatic fibrosis, is a potent EMT inducer. This study aimed to clarify the potential contributions of TGF-β1-induced EMT-like cholangiocyte phenotype to collagen production and cell survival of cholangiocytes in vitro. Mouse cholangiocytes (603B cells) were treated with TGF-β1 and EMT-like phenotype alterations were monitored by morphological changes and expression of EMT-associated genes. Alterations in Col1A1 gene, Col1A1-associated miR-29s, and pro-apoptotic genes were measured in TGF-β1-treated 603B cells. Snail1 knockdown was achieved using shRNA to evaluate the contribution of EMT-associated changes to Col1A1 production and cell survival. We found TGF-β1 treatment induced partial EMT-like phenotype transition in 603B cells in a Snail1-dependent manner. TGF-β1 also stimulated collagen α1(I) expression in 603B cells. However, this induction was not parallel to the EMT-like alterations and independent of Snail1 or miR-29 expression. Cells undergoing EMT-like changes showed a modest down-regulation of multiple pro-apoptotic genes and displayed resistance to TNF-α-induced apoptosis. TGF-β1-induced apoptosis resistance was attenuated in Snail1 knockdown 603B cells. TGF-β1-induced Col1A1 production seems to be independent of EMT-like transition and miR-29 expression. Nevertheless, TGF-β1-induced EMT may contribute to the increased survival capacity of cholangiocytes via modulating the expression of pro-apoptotic genes.


A PDGFRα-Mediated Switch toward CD9high Adipocyte Progenitors Controls Obesity-Induced Adipose Tissue Fibrosis.
Obesity-induced white adipose tissue (WAT) fibrosis is believed to accelerate WAT dysfunction. However, the cellular origin of WAT fibrosis remains unclear. Here, we show that adipocyte platelet-derived growth factor receptor-α-positive (PDGFRα+) progenitors adopt a fibrogenic phenotype in obese mice prone to visceral WAT fibrosis. More specifically, a subset of PDGFRα+ cells with high CD9 expression (CD9high) originates pro-fibrotic cells whereas their CD9low counterparts, committed to adipogenesis, are almost completely lost in the fibrotic WAT. PDGFRα pathway activation promotes a phenotypic shift toward PDGFRα+CD9high fibrogenic cells, driving pathological remodeling and altering WAT function in obesity. These findings translated to human obesity as the frequency of CD9high progenitors in omental WAT (oWAT) correlates with oWAT fibrosis level, insulin-resistance severity, and type 2 diabetes. Collectively, our data demonstrate that in addition to representing a WAT adipogenic niche, different PDGFRα+ cell subsets modulate obesity-induced WAT fibrogenesis and are associated with loss of metabolic fitness.


Cladistic coding of genomic maps.
A new method of genomic maps analysis is described. The purpose of the method is to reconstruct phylogenetic relationships from the genomic organization of taxa. Our approach is based on gene order coding. This coding allows the description of genome topology without a prior hypothesis about evolutionary events and phylogenetic relationships. Different characters are used for each gene: (1) presence/absence, (2) orientation, and (3) relative position. The relative position of a particular gene inside the genome is the pair of genes surrounding it. The relative position character represents all the positions of a gene in the sampled genomes. It is coded as a multistate character. Our coding method has a priori variable cost implications on operators such as inversion, transposition, and gene loss/gain, which we discuss. The overall approach best fits the "duplication, random loss" evolutionary model. The coding method allows the reconstitution of a possible hypothetical common ancestor genome at each node of the tree. This reconstitution is based on the character states' optimization; it comes down to choosing, among all possible optimizations, the optimization compatible with a complete genome topology at each internal node. The multistate coding of gene relative position, which is an undeniable advantage of this method, permits this reconstitution.


Thyroid function in early and late alcohol withdrawal: relationship with aggression, family history, and onset age of alcoholism.
Thyroid dysfunction is a known finding in alcoholism. Most studies have reported the reduction in peripheral thyroid hormones in acute withdrawal and long-term abstinence periods of alcohol dependence. The aim of the present study was to investigate the alterations of free thyroid hormones in early and late withdrawal and their association with aggression, age of onset, and family history of alcoholism. Male inpatients (n = 39; mean age +/- SD: 42.55 +/- 8.02 years) in alcohol withdrawal were compared with healthy men (n = 28; mean age +/- SD: 38.31 +/- 9.26 years). Levels of free thyroxine (fT4), free triiodothyronine, (fT3) and thyrothrophin (TSH) were measured in early (first day) and late (28th day) withdrawal in the patients and only once in the controls. In early withdrawal, levels of thyroid hormones did not differ from those in the controls. In late withdrawal, fT3 and fT4 levels (2.71 +/- 0.56 and 10.80 +/- 1.86 pg/ml) were lower than those of both controls (3.32 +/- 0.41 and 11.95 +/- 1.49 pg/ml, respectively, P < 0.05 in both cases) and patients in early withdrawal (3.18 +/- 0.72 and 12.68 +/- 2.50 pg/ml, respectively, P < 0.05 in both cases). Patients were divided into subgroups according to aggression level, onset age of alcoholism, and family history. While the high-aggression group had lower serum levels of fT3 and fT4 in late withdrawal (2.49 +/- 0.41 and 10.44 +/- 2.15 pg/ml) compared with those of controls (P < 0.05 in both cases), the low-aggression group only had lower serum levels of fT3 in late withdrawal (2.90 +/- 0.62 pg/ml) compared with those of controls (P < 0.05). fT3 and fT4 values in the family history-negative group (2.67 +/- 0.56 and 10.75 +/- 1.88 pg/ml) were lower than those of controls in late withdrawal (P < 0.05 in both cases). Both fT3 and fT4 levels in late withdrawal (2.69 +/- 0.54 and 10.83 +/- 1.96 pg/ml) were decreased in early-onset group compared with those of controls (P < 0.05 in both cases). Decreased free thyroid hormone levels may be a result of heavy alcohol consumption or a trait marker of alcoholism, especially in high-aggressive, early-onset and family history-negative patients.


DNA as patentable subject matter and a narrow framework for addressing the perceived problems caused by gene patents.
Concerns about the alleged harmful effects of gene patents--including hindered research and innovation and impeded patient access to high-quality genetic diagnostic tests--have resulted in overreactions from the public and throughout the legal profession. These overreactions are exemplified by Association for Molecular Pathology v. U.S. Patent and Trademark Office, a 2010 case in the Southern District of New York that held that isolated DNA is unpatentable subject matter under 35 U.S.C. § 101. The problem with these responses is that they fail to adequately consider the role that gene patents and patents on similar biomolecules play in facilitating investment in the costly and risky developmental processes required to transform the underlying inventions into marketable products. Accordingly, a more precisely refined solution is advisable. This Note proposes a narrowly tailored set of solutions to address the concerns about gene patents without destroying the incentives for companies to create and commercialize inventions derived from these and similar patents.


Surgeons' Perspectives of Contralateral Prophylactic Mastectomy.
Contralateral prophylactic mastectomy (CPM) is commonly performed for the treatment of breast cancer, despite its limited oncologic benefit. Little is known about surgeons' perceptions of performing CPM. We hypothesized that a proportion of surgeons would report discomfort with performing CPM, particularly when there is discordance between patients' perceived benefit from CPM and the expected oncologic benefit. A survey was sent to members of the American Society of Breast Surgeons seeking self-reports of surgeons' practice patterns, perceptions, and comfort levels with CPM. Of the 2436 members surveyed, 601 responded (response rate = 24.7 %). The median age of respondents was 52 years, and 59 % of responders were women. The majority (58 %) reported that 80 % of their practice was devoted to the treatment of breast disease. Fifty-seven percent (n = 326) of respondents reported discomfort with performing CPM at some point in their practice. While most surgeons (95 %) were comfortable with CPM on a patient with a deleterious BRCA mutation, only 34 % were comfortable performing CPM on an average-risk patient. The most common reasons reported for surgeon discomfort with CPM were a concern for overtreatment, an unfavorable risk/benefit ratio, and inadequate patient understanding of the anticipated risks and benefits of CPM. Despite the increasing use of CPM for the treatment of breast cancer, many surgeons report discomfort with CPM. Concerns with performing CPM predominantly focus on ambiguities surrounding the oncologic benefit and relative risk of this procedure. Further research is needed to define optimal shared decision-making practices in this area.


Computational correction of copy number effect improves specificity of CRISPR-Cas9 essentiality screens in cancer cells.
The CRISPR-Cas9 system has revolutionized gene editing both at single genes and in multiplexed loss-of-function screens, thus enabling precise genome-scale identification of genes essential for proliferation and survival of cancer cells. However, previous studies have reported that a gene-independent antiproliferative effect of Cas9-mediated DNA cleavage confounds such measurement of genetic dependency, thereby leading to false-positive results in copy number-amplified regions. We developed CERES, a computational method to estimate gene-dependency levels from CRISPR-Cas9 essentiality screens while accounting for the copy number-specific effect. In our efforts to define a cancer dependency map, we performed genome-scale CRISPR-Cas9 essentiality screens across 342 cancer cell lines and applied CERES to this data set. We found that CERES decreased false-positive results and estimated sgRNA activity for both this data set and previously published screens performed with different sgRNA libraries. We further demonstrate the utility of this collection of screens, after CERES correction, for identifying cancer-type-specific vulnerabilities.


Fusion order controls expression level and activity of elastin-like polypeptide fusion proteins.
We have previously developed a method to purify recombinant proteins, termed inverse transition cycling (ITC) that eliminates the need for column chromatography. ITC exploits the inverse solubility phase transition of an elastin-like polypeptide (ELP) that is fused to a protein of interest. In ITC, a recombinant ELP fusion protein is cycled through its phase transition, resulting in separation of the ELP fusion protein from other Escherichia coli contaminants. Herein, we examine the role of the position of the ELP in the fusion protein on the expression levels and yields of purified protein for four recombinant ELP fusion proteins. Placing the ELP at the C-terminus of the target protein (protein-ELP) results in a higher expression level for the four ELP fusion proteins, which also translates to a greater yield of purified protein. The position of the fusion protein also has a significant impact on its specific activity, as ELP-protein constructs have a lower specific activity than protein-ELP constructs for three out of the four proteins. Our results show no difference in mRNA levels between protein-ELP and ELP-protein fusion constructs. Instead, we suggest two possible explanations for these results: first, the translational efficiency of mRNA may differ between the fusion protein in the two orientations and second, the lower level of protein expression and lower specific activity is consistent with a scenario that placement of the ELP at the N-terminus of the fusion protein increases the fraction of misfolded, and less active conformers, which are also preferentially degraded compared to fusion proteins in which the ELP is present at the C-terminal end of the protein.


RON Nuclear Translocation under Hypoxia Potentiates Chemoresistance to DNA Double-Strand Break-Inducing Anticancer Drugs.
Tumor hypoxia is associated with radioresistance, chemoresistance, and metastasis, which eventually lead to cancer progression and a poor patient prognosis. RON [also known as macrophage-stimulating protein receptor (MST1R)] belongs to the c-MET [also known as hepatocyte growth factor receptor (HGFR)] receptor tyrosine kinase (RTK) superfamily. To identify the interaction partners of RON nuclear translocation in response to hypoxia, the nuclear extract of TSGH8301 bladder cancer cells was immunoprecipitated for tandem mass profiling analysis. Nuclear RON interacted with adenosine triphosphate (ATP)-dependent DNA helicase 2 (Ku70) and DNA-dependent protein kinase catalytic subunit (DNA-PKcs) to activate nonhomologous end joining (NHEJ) DNA repair. The interaction was time dependent, extending 3 to 24 hours posthypoxia or until the components had been exposed to the chemotherapeutic drugs doxorubicin and epirubicin. Stable knockdown experiments in vitro suggest the importance of RON for the chemoresistance of cancer cells under hypoxia. In addition, the tyrosine kinase domain of nuclear RON is crucial for interaction with Ku70 under hypoxia. J82 cells transfected with RON showed a survival advantage in the presence of epirubicin and hypoxia. This suggests that nuclear RON activates NHEJ repair by interacting with Ku70/DNA-PKcs and inhibiting RON activity to increase cancer cell chemosensitivity. Mol Cancer Ther; 15(2); 276-86. ©2016 AACR.


Spatio-temporal dynamics of dengue in Brazil: Seasonal travelling waves and determinants of regional synchrony.
Dengue continues to be the most important vector-borne viral disease globally and in Brazil, where more than 1.4 million cases and over 500 deaths were reported in 2016. Mosquito control programmes and other interventions have not stopped the alarming trend of increasingly large epidemics in the past few years. Here, we analyzed monthly dengue cases reported in Brazil between 2001 and 2016 to better characterise the key drivers of dengue epidemics. Spatio-temporal analysis revealed recurring travelling waves of disease occurrence. Using wavelet methods, we characterised the average seasonal pattern of dengue in Brazil, which starts in the western states of Acre and Rondônia, then travels eastward to the coast before reaching the northeast of the country. Only two states in the north of Brazil (Roraima and Amapá) did not follow the countrywide pattern and had inconsistent timing of dengue epidemics throughout the study period. We also explored epidemic synchrony and timing of annual dengue cycles in Brazilian regions. Using gravity style models combined with climate factors, we showed that both human mobility and vector ecology contribute to spatial patterns of dengue occurrence. This study offers a characterization of the spatial dynamics of dengue in Brazil and its drivers, which could inform intervention strategies against dengue and other arboviruses.


Effects of shock waves on oxidative stress, antioxidant enzyme and element levels in kidney of rats.
The aim of this study was to investigate the effect of extracorporeal shock wave lithotripsy (ESWL) on kidney oxidative stress and trace element levels of adult rats. Twelve male Wistar albino rats were divided equally into two groups. First group was used as control. The right-side kidneys of animals in second group were treated with 2,000 18-kV shock waves under anesthesia. Localization of the right kidney was achieved following contrast medium injection through a tail vein under flouroscopy control. The animals were sacrificed 72 h after the ESWL treatment, and the kidneys were taken. Malondialdehyde level was higher in the ESWL group than in the control. Reduced glutathione levels, superoxide dismutase, and glutathione peroxidase activities were lower in the ESWL group than those of the control. Fe, Cu, Pb, Mn, Cd, and Ni levels were lower in the ESWL group than in the control, although Mg level was higher in the ESWL group than in the control. In conclusion, the result of the present study indicated that ESWL treatment produced oxidative stress in the kidney and caused impairments on the antioxidant and trace element levels in the kidneys of rats.


Selection and microevolution of coat pattern are cryptic in a wild population of sheep.
Understanding the maintenance of genetic variation in natural populations is a core aim of evolutionary genetics. Insight can be gained by quantifying selection at the level of the genotype, as opposed to the phenotype. Here, we show that in a natural population of Soay sheep which is polymorphic for coat pattern, recessive genetic variants at the causal gene, agouti signalling protein (ASIP) are associated with reduced lifetime fitness. This was due primarily to a reduction in juvenile survival of uniformly coloured (self-type) sheep, which are homozygous recessive, and occurs despite significantly higher reproductive success in surviving self-type adults. Consistent with their relatively low fitness, we show that the frequency of self-type individuals has declined from 1985 to 2008. Remarkably though, the frequency of the underlying self-allele has increased, because the frequency of heterozygous individuals (who harbour the majority of all self-alleles) has increased. Indeed, the ratio of observed/expected heterozygous individuals has increased during the study, such that there is now a significant excess of heterozygotyes. By employing gene-dropping simulations, we show that microevolutionary trends in the frequency and excess of ASIP heterozygotes are too pronounced to be caused by genetic drift. Studying this polymorphism at the level of phenotype rather than underlying genotype would have failed to detect cryptic fitness differences. We would also have been unable to rule out genetic drift as an evolutionary force driving genetic change. This highlights the importance of resolving the underlying genetic basis of phenotypic variation in explaining evolutionary dynamics.


Characterization of doped amorphous silicon thin films through the investigation of dopant elements by glow discharge spectrometry: a correlation of conductivity and bandgap energy measurements.
The determination of optical parameters, such as absorption and extinction coefficients, refractive index and the bandgap energy, is crucial to understand the behavior and final efficiency of thin film solar cells based on hydrogenated amorphous silicon (a-Si:H). The influence of small variations of the gas flow rates used for the preparation of the p-a-SiC:H layer on the bandgap energy, as well as on the dopant elements concentration, thickness and conductivity of the p-layer, is investigated in this work using several complementary techniques. UV-NIR spectrophotometry and ellipsometry were used for the determination of bandgap energies of four p-a-SiC:H thin films, prepared by using different B(2)H(6) and SiH(4) fluxes (B(2)H(6) from 12 sccm to 20 sccm and SiH(4) from 6 sccm to 10 sccm). Moreover, radiofrequency glow discharge optical emission spectrometry technique was used for depth profiling characterization of p-a-SiC:H thin films and valuable information about dopant elements concentration and distribution throughout the coating was found. Finally, a direct relationship between the conductivity of p-a-SiC:H thin films and the dopant elements concentration, particularly boron and carbon, was observed for the four selected samples.


Profibrogenic chemokines and viral evolution predict rapid progression of hepatitis C to cirrhosis.
Chronic hepatitis C may follow a mild and stable disease course or progress rapidly to cirrhosis and liver-related death. The mechanisms underlying the different rates of disease progression are unknown. Using serial, prospectively collected samples from cases of transfusion-associated hepatitis C, we identified outcome-specific features that predict long-term disease severity. Slowly progressing disease correlated with an early alanine aminotransferase peak and antibody seroconversion, transient control of viremia, and significant induction of IFN-γ and MIP-1β, all indicative of an effective, albeit insufficient, adaptive immune response. By contrast, rapidly progressive disease correlated with persistent and significant elevations of alanine aminotransferase and the profibrogenic chemokine MCP-1 (CCL-2), greater viral diversity and divergence, and a higher rate of synonymous substitution. This study suggests that the long-term course of chronic hepatitis C is determined early in infection and that disease severity is predicted by the evolutionary dynamics of hepatitis C virus and the level of MCP-1, a chemokine that appears critical to the induction of progressive fibrogenesis and, ultimately, the ominous complications of cirrhosis.


Intrauterine insemination or intracervical insemination with cryopreserved donor sperm in the natural cycle: a cohort study.
Does intrauterine insemination in the natural cycle lead to better pregnancy rates than intracervical insemination (ICI) in the natural cycle in women undergoing artificial insemination with cryopreserved donor sperm. In a large cohort of women undergoing artificial insemination with cryopreserved donor sperm, there was no substantial beneficial effect of IUI in the natural cycle over ICI in the natural cycle. At present, there are no studies comparing IUI in the natural cycle versus ICI in the natural cycle in women undergoing artificial insemination with cryopreserved donor sperm. We performed a retrospective cohort study among all eight sperm banks in the Netherlands. We included all women who underwent artificial insemination with cryopreserved donor sperm in the natural cycle between January 2009 and December 2010. We compared time to ongoing pregnancy in the first six cycles of IUI and ICI, after which controlled ovarian stimulation was commenced. Ongoing pregnancy rates (OPRs) over time were compared using life tables. A Cox proportional hazard model was used to compare the chances of reaching an ongoing pregnancy after IUI or ICI adjusted for female age and indication. We included 1843 women; 1163 women underwent 4269 cycles of IUI and 680 women underwent 2345 cycles of ICI with cryopreserved donor sperm. Baseline characteristics were equally distributed (mean age 34.0 years for the IUI group versus 33.8 years for the ICI group), while in the IUI group, there were more lesbian women than in the ICI group (40.6% for IUI compared with 31.8% for ICI). Cumulative OPRs up to six treatment cycles were 40.5% for IUI and 37.9% for ICI. This corresponds with a hazard rate ratio of 1.02 [95% confidence interval (CI) 0.84-1.23] after controlling for female age and indication. Increasing female age was associated with a lower OPR, in both the IUI and ICI groups with a hazard ratio for ongoing pregnancy of 0.94 per year (95% CI 0.93-0.97). This study is prone to selection bias due to its retrospective nature. As potential confounders such as parity and duration of subfertility were not registered, the effect of these potential confounders could not be evaluated. In women inseminated with cryopreserved donor sperm in the natural cycle, we found no substantial benefit of IUI over ICI. A randomized controlled trial with economic analysis alongside, it is needed to allow a more definitive conclusion on the cost-effectiveness of insemination with cryopreserved donor sperm. No funding was used and no conflicts of interest are declared.


Enhancing Gamma Oscillations Restores Primary Motor Cortex Plasticity in Parkinson's Disease.
In humans, γ oscillations in cortical motor areas reflect asynchronous synaptic activity and contribute to plasticity processes. In Parkinson's disease (PD), γ oscillatory activity in the basal ganglia-thalamo-cortical network is altered and the LTP-like plasticity elicited by intermittent theta burst stimulation (iTBS) is reduced in the primary motor cortex (M1). In this study, we tested whether transcranial alternating current stimulation (tACS) delivered at γ frequency promotes iTBS-induced LTP-like plasticity in M1 in PD patients. Sixteen patients (OFF condition) and 16 healthy subjects (HSs) underwent iTBS during γ-tACS (iTBS-γ tACS) and during sham-tACS (iTBS-sham tACS) in two sessions. Motor-evoked potentials (MEPs) evoked by single-pulse transcranial magnetic stimulation and short-interval intracortical inhibition (SICI) were recorded before and after the costimulation. A subgroup of patients also underwent iTBS during β tACS. iTBS-sham tACS facilitated single-pulse MEPs in HSs, but not in patients. iTBS-γ tACS induced a larger MEP facilitation than iTBS-sham tACS in both groups, with similar values in patients and HSs. In patients, SICI improved after iTBS-γ tACS. The effect produced by iTBS-γ tACS on single-pulse MEPs correlated with disease duration, while changes in SICI correlated with Unified Parkinson's Disease Rating Scale Part III scores. The effect of iTBS-β tACS on both single-pulse MEPs and SICI was similar to that obtained in the iTBS-sham tACS session. Our data suggest that γ oscillations have a role in the pathophysiology of the abnormal LTP-like plasticity in PD. Entraining M1 neurons at the γ rhythm through tACS may be an effective method to restore impaired plasticity.SIGNIFICANCE STATEMENT In Parkinson's disease, the LTP-like plasticity of the primary motor cortex is impaired, and γ oscillations are altered in the basal ganglia-thalamo-cortical network. Using a combined transcranial magnetic stimulation-transcranial alternating current stimulation approach (iTBS-γ tACS costimulation), we demonstrate that driving γ oscillations restores the LTP-like plasticity in patients with Parkinson's disease. The effects correlate with clinical characteristics of patients, being more evident in less affected patients and weaker in patients with longer disease duration. These findings suggest that cortical γ oscillations play a beneficial role in modulating the LTP-like plasticity of M1 in Parkinson's disease. The iTBS-γ tACS approach may be potentially useful in rehabilitative settings in patients.


In Vivo Reporter Assays Uncover Changes in Enhancer Activity Caused by Type 2 Diabetes-Associated Single Nucleotide Polymorphisms.
Many single nucleotide polymorphisms (SNPs) associated with type 2 diabetes overlap with putative endocrine pancreatic enhancers, suggesting that these SNPs modulate enhancer activity and, consequently, gene expression. We performed in vivo mosaic transgenesis assays in zebrafish to quantitatively test the enhancer activity of type 2 diabetes-associated loci. Six out of 10 tested sequences are endocrine pancreatic enhancers. The risk variant of two sequences decreased enhancer activity, while in another two incremented it. One of the latter (rs13266634) locates in an SLC30A8 exon, encoding a tryptophan-to-arginine substitution that decreases SLC30A8 function, which is the canonical explanation for type 2 diabetes risk association. However, other type 2 diabetes-associated SNPs that truncate SLC30A8 confer protection from this disease, contradicting this explanation. Here, we clarify this incongruence, showing that rs13266634 boosts the activity of an overlapping enhancer and suggesting an SLC30A8 gain of function as the cause for the increased risk for the disease. We further dissected the functionality of this enhancer, finding a single nucleotide mutation sufficient to impair its activity. Overall, this work assesses in vivo the importance of disease-associated SNPs in the activity of endocrine pancreatic enhancers, including a poorly explored case where a coding SNP modulates the activity of an enhancer.


The Role of NADPH Oxidases (NOXs) in Liver Fibrosis and the Activation of Myofibroblasts.
Chronic liver injury, resulted from different etiologies (e.g., virus infection, alcohol abuse, nonalcoholic steatohepatitis (NASH) and cholestasis) can lead to liver fibrosis characterized by the excess accumulation of extracellular matrix (ECM) proteins (e.g., type I collagen). Hepatic myofibroblasts that are activated upon liver injury are the key producers of ECM proteins, contributing to both the initiation and progression of liver fibrosis. Hepatic stellate cells (HSCs) and to a lesser extent, portal fibroblast, are believed to be the precursor cells that give rise to hepatic myofibroblasts in response to liver injury. Although, much progress has been made toward dissecting the lineage origin of myofibroblasts, how these cells are activated and become functional producers of ECM proteins remains incompletely understood. Activation of myofibroblasts is a complex process that involves the interactions between parenchymal and non-parenchymal cells, which drives the phenotypic change of HSCs from a quiescent stage to a myofibroblastic and active phenotype. Accumulating evidence has suggested a critical role of NADPH oxidase (NOX), a multi-component complex that catalyzes reactions from molecular oxygen to reactive oxygen species (ROS), in the activation process of hepatic myofibroblasts. NOX isoforms, including NOX1, NOX2 and NOX4, and NOX-derived ROS, have all been implicated to regulate HSC activation and hepatocyte apoptosis, both of which are essential steps for initiating liver fibrosis. This review highlights the importance of NOX isoforms in hepatic myofibroblast activation and the progression of liver fibrosis, and also discusses the therapeutic potential of targeting NOXs for liver fibrosis and associated hepatic diseases.


Modelling head losses in granular bed anaerobic baffled reactors at high flows during start-up.
Anaerobic treatment of low strength, high flow wastewaters can only be effective if the technology employed can meet key hydrodynamic requirements: maximising the contact surface area and contact period between the influent substrate and the biomass solids, minimising solid washout from the reactor and minimising the backpressure across the system. Backpressure or head loss is an important hydrodynamic property of gravity-flow packed bed reactors, where the flow is the resultant of frictional forces between the incoming fluid and the solid packing material through which the wastewater percolates. Excessive backpressure caused by high influent flow-rates can reduce the contact surface area and increase the influent head on the upstream side of the biomass bed leading to overflow spills, unstable performance and process failure. This study investigates the factors affecting backpressure across a Granular bed baffled reactor (GRABBR) with variable baffle positions. Experimental results were used to develop a mathematical model to quantify backpressure based on physical characteristics of the seed biomass, fluid-flow conditions and reactor geometry. Results have shown that for a constant flow rate the anaerobic baffled reactor exhibits the least backpressure characteristics when both the upflow and downflow areas are roughly 50% of the total compartmental width.


A neural network to improve dim-light vision? Dendritic fields of first-order interneurons in the nocturnal bee Megalopta genalis.
Using the combined Golgi-electron microscopy technique, we have determined the three-dimensional dendritic fields of the short visual fibres (svf 1-3) and first-order interneurons or L-fibres (L1-4) within the first optic ganglion (lamina) of the nocturnal bee Megalopta genalis. Serial cross sections have revealed that the svf type 2 branches into one adjacent neural unit (cartridge) in layer A, the most distal of the three lamina layers A, B and C. All L-fibres, except L1-a, exhibit wide lateral branching into several neighbouring cartridges. L1-b shows a dendritic field of seven cartridges in layers A and C, dendrites of L2 target 13 cartridges in layer A, L3 branches over a total of 12 cartridges in layer A and three in layer C and L4 has the largest dendritic field size of 18 cartridges in layer C. The number of cartridges reached by the respective L-fibres is distinctly greater in the nocturnal bee than in the worker honeybee and is larger than could be estimated from our previous Golgi-light microscopy study. The extreme dorso-ventrally oriented dendritic field of L4 in M. genalis may, in addition to its potential role in spatial summation, be involved in edge detection. Thus, we have shown that the amount of lateral spreading present in the lamina provides the anatomical basis for the required spatial summation. Theoretical and future physiological work should further elucidate the roles that this lateral spreading plays to improve dim-light vision in nocturnal insects.


Approaching the Transition to Adulthood: Distinctive Profiles of Adolescents Aging out of the Child Welfare System.
The transition to adulthood is marked by new roles and responsibilities in such interrelated domains as education, employment, and family formation. This study investigates the capacity of adolescents on the verge of emancipation from the child welfare system to navigate this transition. To explore heterogeneity in adolescents' preparation for independent living, person-oriented methods are applied to a large, representative sample of youth about to exit foster care. The analysis suggests four subpopulations defined by distinctive profiles on indicators reflecting multiple domains of life experience. Identifying the particular needs and challenges of subpopulations has implications for efforts to match adolescents aging out of the child welfare system with appropriate services.


Reverse-micelle model: pH, electromagnetic field and inhibitor enzyme interaction.
The reverse micelle is one of many models thought to have properties more nearly resembling the biological cellular environment, than does the traditional dilute-solution biochemical reaction system. In order to evaluate the results of EMF perturbation of enzyme-catalyzed reactions, the description of the AOT reverse-micelle model, with respect to its internal pH, effect of chemical inhibitors, temperature, and electromagnetic-field perturbation has herein been extended. Acetylcholinesterase and NADPH cytochrome-P450 reductase, reacting within the AOT reverse-micelle, exhibit a temperature vs. activity profile equivalent to the same reaction in a buffered dilute-solution environment. In reverse micelles, some inhibitors of AChE (propidium, and d-tubocurarine) have much less effect upon indophenol-acetate hydrolysis than they do in a dilute solution environment. Other inhibitors act in the same manner within the structured environment of the reverse micelle as in the conventional dilute solution reaction model. These differences are explicable in terms of mechanism of action of the individual inhibitors. Perturbation by low-intensity microwave fields has a similar inhibitory effect upon dilute-solution reactions, as those in the 'low-water-activity' environment of the reverse micelle. However, the interactions between physical and chemical perturbants are differently limited by the structure of the aqueous phase of the reverse micelle. pH of the 'internal' reverse-micelle environment is a function of the availability of H-ions supplied by system components. Use of indicator dyes show that the low-molarity buffers which are compatible with reverse-micelle stability, are often insufficient to maintain a constant pH. Too, in the reverse micelle, reaction rate, for proton yielding reactions, is dramatically greater than the rate of the same reaction in dilute solution at the same acidic pH.


Controlled Cre/loxP site-specific recombination in the developing brain in medaka fish, Oryzias latipes.
Genetic mosaic techniques have been used to visualize and/or genetically modify a neuronal subpopulation within complex neural circuits in various animals. Neural populations available for mosaic analysis, however, are limited in the vertebrate brain. To establish methodology to genetically manipulate neural circuits in medaka, we first created two transgenic (Tg) medaka lines, Tg (HSP:Cre) and Tg (HuC:loxP-DsRed-loxP-GFP). We confirmed medaka HuC promoter-derived expression of the reporter gene in juvenile medaka whole brain, and in neuronal precursor cells in the adult brain. We then demonstrated that stochastic recombination can be induced by micro-injection of Cre mRNA into Tg (HuC:loxP-DsRed-loxP-GFP) embryos at the 1-cell stage, which allowed us to visualize some subpopulations of GFP-positive cells in compartmentalized regions of the telencephalon in the adult medaka brain. This finding suggested that the distribution of clonally-related cells derived from single or a few progenitor cells was restricted to a compartmentalized region. Heat treatment of Tg(HSP:Cre x HuC:loxP-DsRed-loxP-GFP) embryos (0-1 day post fertilization [dpf]) in a thermalcycler (39°C) led to Cre/loxP recombination in the whole brain. The recombination efficiency was notably low when using 2-3 dpf embyos compared with 0-1 dpf embryos, indicating the possibility of stage-dependent sensitivity of heat-inducible recombination. Finally, using an infrared laser-evoked gene operator (IR-LEGO) system, heat shock induced in a micro area in the developing brains led to visualization of clonally-related cells in both juvenile and adult medaka fish. We established a noninvasive method to control Cre/loxP site-specific recombination in the developing nervous system in medaka fish. This method will broaden the neural population available for mosaic analyses and allow for lineage tracing of the vertebrate nervous system in both juvenile and adult stages.


Structure and dynamics of a human myelin protein P2 portal region mutant indicate opening of the β barrel in fatty acid binding proteins.
Myelin is a multilayered proteolipid sheath wrapped around selected axons in the nervous system. Its constituent proteins play major roles in forming of the highly regular membrane structure. P2 is a myelin-specific protein of the fatty acid binding protein (FABP) superfamily, which is able to stack lipid bilayers together, and it is a target for mutations in the human inherited neuropathy Charcot-Marie-Tooth disease. A conserved residue that has been proposed to participate in membrane and fatty acid binding and conformational changes in FABPs is Phe57. This residue is thought to be a gatekeeper for the opening of the portal region upon ligand entry and egress. We performed a structural characterization of the F57A mutant of human P2. The mutant protein was crystallized in three crystal forms, all of which showed changes in the portal region and helix α2. In addition, the behaviour of the mutant protein upon lipid bilayer binding suggested more unfolding than previously observed for wild-type P2. On the other hand, membrane binding rendered F57A heat-stable, similarly to wild-type P2. Atomistic molecular dynamics simulations showed opening of the side of the discontinuous β barrel, giving important indications on the mechanism of portal region opening and ligand entry into FABPs. The results suggest a central role for Phe57 in regulating the opening of the portal region in human P2 and other FABPs, and the F57A mutation disturbs dynamic cross-correlation networks in the portal region of P2. Overall, the F57A variant presents similar properties to the P2 patient mutations recently linked to Charcot-Marie-Tooth disease. Our results identify Phe57 as a residue regulating conformational changes that may accompany membrane surface binding and ligand exchange in P2 and other FABPs.


Viruses of the Nahant Collection, characterization of 251 marine Vibrionaceae viruses.
Viruses are highly discriminating in their interactions with host cells and are thought to play a major role in maintaining diversity of environmental microbes. However, large-scale ecological and genomic studies of co-occurring virus-host pairs, required to characterize the mechanistic and genomic foundations of virus-host interactions, are lacking. Here, we present the largest dataset of cultivated and sequenced co-occurring virus-host pairs that captures ecologically representative fine-scale diversity. Using the ubiquitous and ecologically diverse marine Vibrionaceae as a host platform, we isolate and sequence 251 dsDNA viruses and their hosts from three time points within a 93-day time-series study. The virus collection includes representatives of the three Caudovirales tailed virus morphotypes, a novel family of nontailed viruses, and the smallest (10,046 bp) and largest (348,911 bp) Vibrio virus genomes described. We provide general characterization and annotation of the viruses and describe read-mapping protocols to standardize genome presentation. The rich ecological and genomic contextualization of hosts and viruses make the Nahant Collection a unique platform for high-resolution studies of environmental virus-host infection networks.


Simple contact mechanics model of the vertebrate cartilage.
We study a simple contact mechanics model of the vertebrate cartilage, which includes (bulk) osmotic effects. The surface roughness power spectrum of a pig cartilage is obtained from the measured surface topography. Using the Reynolds equations with fluid flow factors, calculated using the Persson contact mechanics theory and the Bruggeman effective medium theory, we show how the area of contact and the average interfacial separation change with time. We found that in most cases the contact area percolates, resulting in islands of confined fluid which carry most of the external load. Most importantly, we find that the pressure in the area of real contact is nearly independent of the external load, and well below 1 MPa. This allows the surfaces in the area of "real contact", to be separated (at nanometer range separation distance) by osmotic repulsion, resulting in a very small (breakloose) friction force observed even after a long time of stationary contact.


A new genus of Rhinebothriidea from species of Psammobatis (Rajiformes: Arhynchobatidae) off Argentina.
Two species of a new genus of Rhinebothriidea have been collected from species of Psammobatis Günther along the coast of Argentina. Semiorbiseptum gen. nov. is considered a rhinebothriidean for the presence of facially loculated bothridia borne on stalks and cirrus covered with spinitriches. The genus is unique in the arrangement of loculi and septa on the distal bothridial surface, including semicircular septa in the middle region. Despite the anterior-posterior orientation of bothridia, Semiorbiseptum gen. nov. fits well in the description of the family Rhinebothriidae by lacking a myzorhynchus at the adult stage and postvaginal testes, and by having vitelline follicles that are not interrupted at the level of the ovary. The two new species differ from each other in the number of facial bothridial loculi and in the microthrix pattern on the proximal bothridial surface. Unlike the majority of rhinebothrideans in the marine environment, which show high specificity for their definitive host, Semiorbiseptum mariae sp. nov. was found in two species of Psammobatis, the smallthorn sand skate Psammobatis rudis Günther and shortfin sand skate Psammobatis normani McEachran. These species have a sympatric distribution, similar to the hosts of most rhinebothrideans that parasitize more than one host. On the contrary, Semiorbiseptum alfredoi sp. nov. was found only in P. normani. This study shows that some of the characters previously proposed as possible synapomorphies for the order have interspecific variability or can be equivocally interpreted (e.g. the entrance of the vas deferens into the cirrus sac, bothridial stalk shape).


Financial Considerations Associated With a Fourth Year of Residency Training in Family Medicine: Findings From the Length of Training Pilot Study.
The feasibility of funding an additional year of residency training is unknown, as are perspectives of residents regarding related financial considerations. We examined these issues in the Family Medicine Length of Training Pilot. Between 2013 and 2019, we collected data on matched 3-year and 4-year programs using annual surveys, focus groups, and in-person and telephone interviews. We analyzed survey quantitative data using descriptive statistics, independent samples t test, Fisher's Exact Test and χ2. Qualitative analyses involved identifying emergent themes, defining them and presenting exemplars. Postgraduate year (PGY)-4 residents in 4-year programs were more likely to moonlight to supplement their resident salaries compared to PGY-3 residents in three-year programs (41.6% vs 23.0%; P=.002), though their student debt load was similar. We found no differences in enrollment in loan repayment programs or pretax income. Programs' descriptions of financing a fourth year as reported by the program director were limited and budget numbers could not be obtained. However, programs that required a fourth year typically reported extensive planning to determine how to fund the additional year. Programs with an optional fourth year were budget neutral because few residents chose to undertake an additional year of training. Resources needed for a required fourth year included resident salaries for the fourth year, one additional faculty, and one staff member to assist with more complex scheduling. Residents' concerns about financial issues varied widely. Adding a fourth year of training was financially feasible but details are local and programs could not be compared directly. For programs that had a required rather than optional fourth year much more financial planning was needed.


Tilted seat position for non-ambulant individuals with neurological and neuromuscular impairment: a systematic review.
To determine the effects of tilt-in-space seating on outcomes for people with neurological or neuromuscular impairment who cannot walk. Search through electronic databases (MEDLINE, Embase, CINAHL, AMED). Discussions with researchers who are active in field. Selection criteria included interventional studies that investigated the effects of seat tilt on outcome or observational studies that identified outcomes for those who had used tilt-in-space seating in populations with neurological or neuromuscular impairments. Two reviewers independently selected trials for inclusion, assessed quality and extracted data. Nineteen studies were identified which fulfilled the selection criteria. Seventeen of these were essentially before-after studies investigating the immediate effects of tilting the seating. All studies looked at populations with neurological impairment, and most were on children with cerebral palsy (n=8) or adults with spinal cord injury (n=8). REVIEWER'S CONCLUSION: Posterior tilt can reduce pressures at the interface under the pelvis.


Birth weight, gestational age, and blood pressure: Early life management strategy and population health perspective.
The incidence of hypertension (HTN) is rising worldwide with an estimated prevalence of 22%, 7.5 million deaths (12.8%). It is a major risk factor for coronary heart diseases and hemorrhagic strokes. In Oman, the crude prevalence of HTN was 33.1%, whereas the age-adjusted prevalence was 38.3%. Among Gulf Cooperation Countries, 47.2% of the individuals were hypertensive, and women were more likely to have HTN than men. Similarly, the prevalence of low-birth-weight (LBW) is also rising globally with the more prevalent incidence in developing countries reaching almost a rate just lower than 20.0/100 births. In Oman, the prevalence of LBW was 4.2% in 1980, which doubled (8.1%) in 2000 and has shown a slow but steady increase reaching 10.2% in 2013. LBW term is the most commonly used surrogate measure of intrauterine growth retardation and has been related to increased cardiovascular mortality, due to increased risk of cardiovascular risk factors, including blood pressure (BP), diabetes, cholesterol level, and other risk factors. The epidemiologic evidence clearly points to an inverse association between birth weight and many hemodynamic cardiovascular risk markers. Possible mechanisms operating in fetal life that might determine BP include the structural development of resistance arteries, the setting of hormone levels, and nephron endowment. Retarded fetal growth leads to permanently reduced cell numbers in the kidney. Patients with high BP had almost 50% less number of glomeruli compared to that of the normotensive individuals, and subsequent accelerated growth may lead to excessive metabolic demand on this limited cell mass. It is not merely a reduced nephron number that is responsible for HTN, but compensatory maladaptive changes that occur internally when nephrogenesis is compromised. The likelihood of an adverse outcome is greatly amplified in those born with LBW who later develop obesity or an increased ponderal index.


Nosema cuiculi: in vitro isolation.
A mammalian microsporidian was propagated for the first time in vitro with rabbit choroid plexus monolayer cultures. Tentative classification as Nosema cuniculi (Levaditi, 1923) was established by morphologic and tinctorial characteristics and by extrusion of sintgle polar filaments with attached sporoplasin from living spores. The method permits propagation of large numbers of organisms in putre cultutre.


"That Can't Be!": Perceptions of HIV and Hepatitis C Screening during Admission to an Acute Care Surgery Service.
A large number of patients live with undiagnosed HIV and/or hepatitis C despite broadened national screening guidelines. European studies, however, suggest many patients falsely believe they have been screened during a prior hospitalization. This study aims to define current perceptions among trauma and emergency general surgery (EGS) patients regarding HIV and hepatitis C screening practices. Prospective survey administered to adult (>18 years old) acute care surgery service (trauma and EGS) patients at a Level 1 academic trauma center. The survey consisted of 13 multiple choice questions: demographics, whether admission tests included HIV and hepatitis C at index and prior hospital visits and whether receiving no result indicated a negative result, prior primary care screening. Response percentages calculated in standard fashion. One hundred and twenty-five patients were surveyed: 80 trauma and 45 EGS patients. Overall, 32% and 29.6% of patients believed they were screened for HIV and hepatitis C at admission. There was no significant difference in beliefs between trauma and EGS. Sixty-eight percent of patients had a hospital visit within 10 years of these, 49.3% and 44.1% believe they had been screened for HIV and hepatitis C. More EGS patients believed they had a prior screen for both conditions. Among patients who believed they had a prior screen and did not receive any results, 75.9% (HIV) and 80.8% (hepatitis C) believed a lack of results meant they were negative. Only 28.9% and 23.6% of patients had ever been offered outpatient HIV and hepatitis C screening. A large portion of patients believe they received admission or prior hospitalization HIV and/or hepatitis C screening and the majority interpreted a lack of results as a negative diagnosis. Due to these factors, routine screening of trauma/EGS patients should be considered to conform to patient expectations and national guidelines, increase diagnosis and referral for medical management, and decrease disease transmission.


An Examination of the Differential Effects of the Modern Epidemiological Transition on Cranial Morphology in the United States and Portugal.
This research examines the pattern of secular change in the cranial morphology of two populations experiencing the epidemiological transition associated with decreased mortality rates in children, followed by declines in infant mortality and subsequent increases in adult longevity. The two samples examined in this study come from US and Portuguese individuals. The epidemiological transition occurred at different times in the United States and Portugal, with Portugal entering into the transition later than the United States. The results of the study show that the US and Portuguese samples experienced significant changes in cranial morphology during the approximately 150 years under study. In all of the samples the cranial base morphology changes significantly over time. However, the pattern of change in the US and Portuguese samples varies in the other regions of the crania. The US samples exhibit significant changes associated with the posterior cranial fossa, which experiences the greatest growth during the fetal period and the first year of life. Conversely, in the Portuguese samples the region of the cranium that shows the greatest change is in the face and lateral cranial base, which experiences the greatest growth from three to nine years. This differential pattern may reflect differences in changing mortality patterns in the two countries. During the period under study the United States had already proceeded through the early stages of the epidemiological transition, and improvements in the juvenile mortality and juvenile growth had occurred previously. Subsequently, the United States experienced significant declines in infant mortality, and the regions of the crania that exhibit the greatest changes occur in area with maximum growth velocity under one year. However, Portugal entered into the epidemiological transition later than the United States and therefore the greatest changes in growth occurred during the juvenile period, which is reflected in the adult morphology in this group. This study demonstrates the utility of variation in growth patterns in different cranial regions to document changes in the demographic parameters in two different populations.


Evolution of the R2 retrotransposon ribozyme and its self-cleavage site.
R2 is a non-long terminal repeat retrotransposon that inserts site-specifically in the tandem 28S rRNA genes of many animals. Previously, R2 RNA from various species of Drosophila was shown to self-cleave from the 28S rRNA/R2 co-transcript by a hepatitis D virus (HDV)-like ribozyme encoded at its 5' end. RNA cleavage was at the precise 5' junction of the element with the 28S gene. Here we report that RNAs encompassing the 5' ends of R2 elements from throughout its species range fold into HDV-like ribozymes. In vitro assays of RNA self-cleavage conducted in many R2 lineages confirmed activity. For many R2s, RNA self-cleavage was not at the 5' end of the element but at 28S rRNA sequences up to 36 nucleotides upstream of the junction. The location of cleavage correlated well with the types of endogenous R2 5' junctions from different species. R2 5' junctions were uniform for most R2s in which RNA cleavage was upstream in the rRNA sequences. The 28S sequences remaining on the first DNA strand synthesized during retrotransposition are postulated to anneal to the target site and uniformly prime second strand DNA synthesis. In species where RNA cleavage occurred at the R2 5' end, the 5' junctions were variable. This junction variation is postulated to result from the priming of second strand DNA synthesis by chance microhomologies between the target site and the first DNA strand. Finally, features of R2 ribozyme evolution, especially changes in cleavage site and convergence on the same active site sequences, are discussed.


Sensitivity of quantitative microbial risk assessments to assumptions about exposure to multiple consumption events per day.
Quantitative microbial risk assessments (QMRAs) of contaminated drinking water usually assume the daily intake volume is consumed once a day. However, individuals could consume water at multiple time points over 1 day, so the objective was to determine if the number of consumption events per day impacted the risk of infection from Campylobacter jejuni during short-term contamination events. A probabilistic hydraulic and risk model was used to evaluate the impact of multiple consumption events as compared to one consumption event on the health risk from the intake of contaminated tap water. The fraction of the population that experiences greater than 10(-4) risk of infection per event at the median dose was 6.8% (5th-95th percentile: 6.5-7.2%) for one consumption event per day, 18.2% (5th-95th: 17.6-18.7%) for three consumption events per day, and 19.8% (5th-95th: 14.0-24.4%) when the number of consumption events varied around 3.49 events/day. While the daily intake volume remained consistent across scenarios, the results suggest that multiple consumption events per day increases the probability of infection during short-term, high level contamination events due to the increased coincidence of a consumption event during the contamination peak. Therefore, it will be important to accurately characterize this parameter in drinking water QMRAs.


In situ studies of materials for high temperature CO2 capture and storage.
Carbon capture and storage (CCS) offers a possible solution to curb the CO2 emissions from stationary sources in the coming decades, considering the delays in shifting energy generation to carbon neutral sources such as wind, solar and biomass. The most mature technology for post-combustion capture uses a liquid sorbent, amine scrubbing. However, with the existing technology, a large amount of heat is required for the regeneration of the liquid sorbent, which introduces a substantial energy penalty. The use of alternative sorbents for CO2 capture, such as the CaO-CaCO3 system, has been investigated extensively in recent years. However there are significant problems associated with the use of CaO based sorbents, the most challenging one being the deactivation of the sorbent material. When sorbents such as natural limestone are used, the capture capacity of the solid sorbent can fall by as much as 90 mol% after the first 20 carbonation-regeneration cycles. In this study a variety of techniques were employed to understand better the cause of this deterioration from both a structural and morphological standpoint. X-ray and neutron PDF studies were employed to understand better the local surface and interfacial structures formed upon reaction, finding that after carbonation the surface roughness is decreased for CaO. In situ synchrotron X-ray diffraction studies showed that carbonation with added steam leads to a faster and more complete conversion of CaO than under conditions without steam, as evidenced by the phases seen at different depths within the sample. Finally, in situ X-ray tomography experiments were employed to track the morphological changes in the sorbents during carbonation, observing directly the reduction in porosity and increase in tortuosity of the pore network over multiple calcination reactions.


Role of stereotactic radiosurgery for multiple (>4) brain metastases.
Stereotactic radiosurgery (SRS) is a very important treatment option for patients with brain metastases. Prospective data supports the use of SRS for patients with 1-4 brain metastases. Although whole brain radiation therapy (WBRT) has been an effective adjunct to SRS and surgery and has also provided effective palliation for many patients with brain metastases, the potential side effects especially neurocognitive function decline has increased the use of SRS alone even for patients with multiple (>4) brain metastases despite data that suggests that tumor progression is worse than the potential neurocognitive effects of WBRT. In addition, current stereotactic radiosurgery machines and techniques allow the delivery of SRS to multiple lesions in an efficient manner. As a result, the optimal management of multiple brain metastases (>4) is becoming more contentious given the lack of prospective data. Until further data is available, a multidisciplinary team of neurosurgeons, radiation oncologists, medical oncologists and medical physicists should work closely together to implement individualized treatment for patients with multiple brain metastases. This paper will review some of the institutional, multi-institutional and randomized trials of SRS for patients with brain metastases, and review the outcomes for patients with multiple (>4) brain metastases treated by SRS and the associated costs.


Ispagula powder: an allergen in the work environment.
Bulk laxatives based on ispagula powder have given no known allergic side effects when administered perorally. Twenty-seven out of 64 workers in a pharmaceutical factory developed rhinitis, conjunctivitis or asthma when engaged in the packing of ispagula husks, an operation which gives rise to dust. The symptoms are accounted for by a type 1 allergy acquired from ispagula husks. This allergy can be demonstrated by intracutaneous and provocation tests. Occupational health measures were taken which reduced the dust content to a minimum; thereafter only one new case of sensitization was recorded during the next 24 months of observation. Five similar cases were reported among nursing staff exposed to the dust of ispagula while administering doses of bulk laxative.


Tailoring lanthanide doping in perovskite CaTiO3 for luminescence applications.
Perovskite oxide materials have been attracting significant attention due to their rich physical and chemical properties. With its proven stability and bio-compatibility, we suggest the lanthanide-doped perovskite to be a promising material for biological luminescence applications. Here, taking CaTiO3 as a concrete example, we systematically investigate its doping properties using first-principles computational methods. We determine the conditions allowing the growth of CaTiO3 against various competing phases. We obtain the formation energies of various intrinsic point defects in the material. The doping configuration and the charge state of the lanthanide dopants are determined. We find that for heavier elements in the lanthanide family, the substitution at the Ca site is favored under p-type growth conditions and tends to be trivalent, whereas the substitution at the Ti site is favored under n-type growth conditions and tends to be divalent. And for lighter elements in the family, the substitution at the Ca site is more favored for most cases and the dopant is more likely to be trivalent. By tuning the growth conditions, one could control the valence state of the lanthanide dopant, which in turn controls the luminescence spectra. We collect and identify the emission peaks in the infrared biological window, based on which possible doping schemes are suggested for bio-labeling and imaging applications.


[Active interventions in hypercholesteroloemia patients with high cardivascular risk in primary care].
Hypercholesterolemia is a major modifiable risk factors for cardiovascular disease (CVD). Its reduction reduces morbidity and mortality from ischemic heart disease and CVD in general, primary prevention and secondary prevention especially. To determine whether a notarized and intensive clinical practice can overcome inertia and achieve the therapeutic goal (OT) LDL-C <100 mg / dL in high-risk patients attended in Primary Care (PC) in our country. epidemiological, prospective, multicenter study conducted in centers of different ACs By AP consecutive sampling 310 patients at high cardiovascular risk (diabetic or established CVD) previously treated with statins, which did not reach the OT included c-LDL. The study subjects had a mean age of 65.2 years, of which 60.32% were male. The 41.64% had a previous EVC, acute myocardial infarction (20.33%), angina (16.07%), stroke / TIA (9.19%), arthropathy (5.25%), diabetes (70 , 87%), hypertension (71.01%), and abdominal obesity (69.62%). The 43.57% (95% CI: 37,21; 50,08) of patients who performed the 2nd visit (241) got the OT. 62.50% (95% CI: 55.68, 68.98) of those who took the 3rd (216) got the OT. Finally, 77.56% (95% CI: 72.13, 83.08) patients who performed the last visit (205) got the OT. Throughout the study there was a reduction in LDL-C levels from 135.6 mg / dL at baseline, 107.4 mg / dL in the 2nd visit, 97.3 mg / dL in the 3rd visit, up to 90.7 mg / dL at the final visit (p <0.0001) The increase in HDL-C from baseline (50.9 mg / dL) and final (53.6 mg / dL) was also significant (p = 0.013). The reassessment and intensification of treatment in patients at high cardiovascular risk treated in primary care, applying the indications of the guides, achieves the OT in more than three quarters of the previously uncontrolled within half a year. These results should encourage us to overcome the therapeutic inertia in the control of CVD by early and energetic performance against hypercholesterolemia.


Plant-parasitic Nematodes in the Waimanalo, Hawaii Irrigation System from Watershed to Farm.
Nematode occurrence at specific locations throughout a water catchment-irrigation system was determined. Soil samples were collected from five water source locations on the slopes of Olomana Mountain and Maunawili Valley and from about 40 plant species on 18 farms (56 ha of 480 ha irrigated by the reservoir). Water was sampled from the catchment reservoir at 0.3 m, 9 m, and 18 m (bottom). A farm irrigated with potable water was sampled and compared to areas of the same farm irrigated from the reservoir. Nematodes present in soil from the mountain and farms were root-knot (Meloidogyne spp.), lesion (Pratylenchus spp.), reniform (Rotylenchulus reniformis), stunt (Tylenchorhynchus sp.), ring (Criconema spp.), dagger (Xiphinema sp.), spiral (Helicotylenchus sp.), Tylenchus sp., Aphelenchus sp., and pin (Paratylenchus sp.) nematodes. The economically important genera Rotylenchulus, Meloidogyne, and Pratylenchus occurred in very low numbers (10, 41, and 10/250 cm(3) soil, respectively) and in low frequency (10%, 25%, and 8% of the samples, respectively) in the mountain samples compared with high numbers (170-895/250 cm(3) soil) from farms. Frequency of occurrence over all farms was near 40% for Meloidogyne and 80% for Rotylenchulus. No nematodes were detected in water from the reservoir. One sample from the outlets contained two specimens of plant-parasitic nematodes. The population densities of nematodes were not different between the soil samples collected from crops irrigated by potable or reservoir water.


Split-EGFP screens for the detection and localisation of protein-protein interactions in living yeast cells.
Proteomics aims to identify and classify the proteins present in a particular cell or tissue. However, we know that proteins rarely function alone and knowledge of which proteins interact with which other proteins is vital if we wish to understand how cells work. The budding yeast, Saccharomyces cerevisiae, is a well-established model for studying protein-protein interactions, and a number of methods have been developed to do this. A method for the in vivo detection and localisation of interacting pairs of proteins in living yeast cells is presented. The method relies on the ability of fragments of enhanced green fluorescent protein (EGFP) to reassemble if brought into close proximity. The reassembled EGFP regains the ability to fluoresce, and this fluorescence can be detected providing evidence of interaction and information about its location. S. cerevisiae is an ideal organism to apply this method to due to the relative ease with which its genome can be manipulated. The method described enables the modification of S. cerevisiae genes at the 3'-end with DNA encoding fragments of EGFP. Consequently, the expression levels of the proteins are unlikely to be affected and thus the method is unlikely to result in false positives. In addition to the protocol for labelling and detection of interacting pairs of yeast proteins, methods for simple tests for the effects of the labelling on the organism's function are presented.


Monolithic 626 nm single-mode AlGaInP DBR diode laser.
Single-mode lasers below 630 nm are still realized using complex laser systems. We present distributed Bragg reflector (DBR) ridge waveguide lasers (RWL) based on AlGaInP. When packaged into sealed TO-3 housings and cooled internally to about 0°C the DBR-RWL emit more than 50 mW at a wavelength of 626.0 nm into a nearly diffraction-limited single longitudinal mode with a spectral width below 1 MHz. These new monolithic diode lasers have the potential to drastically miniaturize existing set-ups e.g. for quantum information processing.


Is there any value to hepatitis B virus genotype analysis?
Hepatitis B may cause a varying spectrum of diseases ranging from an asymptomatic or mild anicteric acute illness, to severe or fulminant hepatitis. Similarly, the outcome of chronic hepatitis B is variable. Viral factors associated with outcome of chronic hepatitis B virus (HBV) infection include hepatitis B e antigen status, HBV DNA, genotype, and HBV variants. HBV genotypes and subgenotypes have been associated with differences in clinical and virological characteristics, indicating that they may play a role in the virus-host relationship. A total of ten hepatitis B virus genotypes have been defined with a distinct geographical distribution. Hitherto, genotypes A, B, C and D have been studied most extensively. The HBV genotype appears to influence not only the natural history of HBV related liver disease but also the response to HBV treatment. HBV genotypes are also linked with both core promoter and BCP mutations. Progression to chronic infection appears to occur more frequently following acute infection with genotypes A and D than with the other studied genotypes. Genotypes A and B appear to have higher rates of spontaneous HBeAg seroconversion. More advanced liver disease and progression to HCC is more often seen in chronic infection with genotypes C and D in contrast to genotypes A and B. More specifically, genotypes A1, C, B2-B5 and H appear to be associated with more serious complications than genotypes A2, B1 and B6. These observations suggest important pathogenic differences between HBV genotypes. Genotypes A and B have higher response rates to interferon based therapy than genotypes C and D. Knowledge of HBV genotype enables clinicians to identify those patients at increased risk of disease progression whilst aiding the selection of appropriate antiviral therapy. Genotyping and monoclonal subtyping can provide useful information for epidemiological studies. In conclusion, genotyping of chronic HBV infections can help practicing physicians identify those at risk of disease progression and determine optimal anti-viral therapy.


Power loss, modal noise and distortion due to microbending of optical fibers.
Theoretical and experimental investigations are described for determining the transmission characteristics of a multimode fiber with microbending for coherent and partially coherent illumination. The measured values of the average excess power loss are shown to be in close agreement with the theory. Also, an estimate of the excess transient loss due to mode coupling is found to be in good agreement with previously published data. Mode-mode interference is shown to be the cause of temporal fluctuations in the microbending loss, from which expressions for modal noise and baseband/subcarrier nonlinearity are derived on a statistical basis. For a given overall loss, the results show that many uniformly distributed small amplitude microbends cause much less modal noise and distortion than a few large amplitude microbends.


Humans can discriminate more than 1 trillion olfactory stimuli.
Humans can discriminate several million different colors and almost half a million different tones, but the number of discriminable olfactory stimuli remains unknown. The lay and scientific literature typically claims that humans can discriminate 10,000 odors, but this number has never been empirically validated. We determined the resolution of the human sense of smell by testing the capacity of humans to discriminate odor mixtures with varying numbers of shared components. On the basis of the results of psychophysical testing, we calculated that humans can discriminate at least 1 trillion olfactory stimuli. This is far more than previous estimates of distinguishable olfactory stimuli. It demonstrates that the human olfactory system, with its hundreds of different olfactory receptors, far outperforms the other senses in the number of physically different stimuli it can discriminate.


Mouse/human chimeric Me1-14 antibody: genomic cloning of the variable region genes, linkage to human constant region genes, expression, and characterization.
Murine monoclonal antibody Me1-14, which recognizes an epitope on chondroitin proteoglycan sulfate expressed in malignant glioma and melanoma, has been used for radioimmunolocalization and therapy both in animal models and in patients. Here, we report the generation, characterization, and in vivo biodistribution of mouse/human chimeric Me1-14. Rearranged immunoglobulin genes from the Me1-14 hybridoma were identified by Southern blot analysis. Putative rearranged light- and heavy-chain genes were cloned from Lambda-ZapII Me1-14 genomic libraries and sequenced for nucleotide analysis. One of the putative heavy-chain Eco RI fragments (3.5 kb) had all the features of an intact variable region, including a functional leader sequence, in-frame V-D and D-J junctions, and cysteines 22 and 92. The deduced amino acid sequence from the heavy-chain variable region gene showed considerable homology with the invariant protein sequence of the mouse heavy-chain subgroup IIIB. Like the heavy-chain gene, one of the putative rearranged kappa-chain Hind III fragments (4 kb) had all of the characteristics of the functional variable region, and the deduced amino acid sequence showed homology to the invariant sequence of kappa-chain group V. The variable region genes for heavy- and light-chains were linked to human constant region exons in the expression vectors at the unique sites and cotransfected into mouse SP2/0 cells. The production level of chimeric Me1-14 from ascites in the highest expressing transfectoma was 1.8 mg/ml. The chimeric Me1-14 antibody exhibited the same specificity and similar affinity as that of parent Me1-14. Direct comparison of radioiodinated chimeric and murine Me1-14 in paired-label biodistribution analysis in subcutaneous xenograft-bearing mice showed higher tumor-to-normal organ ratios for chimeric Me1-14 IgG2, suggesting that this chimeric Me1-14 may be potentially useful in vivo for diagnostic and therapeutic purposes in patients.


Sediment and turbidity associated with offshore dredging increase coral disease prevalence on nearby reefs.
In recent decades, coral reef ecosystems have declined to the extent that reefs are now threatened globally. While many water quality parameters have been proposed to contribute to reef declines, little evidence exists conclusively linking specific water quality parameters with increased disease prevalence in situ. Here we report evidence from in situ coral health surveys confirming that chronic exposure to dredging-associated sediment plumes significantly increase the prevalence of white syndromes, a devastating group of globally important coral diseases. Coral health surveys were conducted along a dredging-associated sediment plume gradient to assess the relationship between sedimentation, turbidity and coral health. Reefs exposed to the highest number of days under the sediment plume (296 to 347 days) had two-fold higher levels of disease, largely driven by a 2.5-fold increase in white syndromes, and a six-fold increase in other signs of compromised coral health relative to reefs with little or no plume exposure (0 to 9 days). Multivariate modeling and ordination incorporating sediment exposure level, coral community composition and cover, predation and multiple thermal stress indices provided further confirmation that sediment plume exposure level was the main driver of elevated disease and other compromised coral health indicators. This study provides the first evidence linking dredging-associated sedimentation and turbidity with elevated coral disease prevalence in situ. Our results may help to explain observed increases in global coral disease prevalence in recent decades and suggest that minimizing sedimentation and turbidity associated with coastal development will provide an important management tool for controlling coral disease epizootics.


Structural studies of the cell wall polysaccharides from three strains of Lactobacillus helveticus with different autolytic properties: DPC4571, BROI, and LH1.
Lactobacillus helveticus is traditionally used in dairy industry as a starter or an adjunct culture for manufacture of cheese and some types of fermented milk. Its autolysis releases intracellular enzymes which is a prerequisite for optimum cheese maturation, and is known to be strain dependent. Autolysis is caused by an enzymatic hydrolysis of the cell wall peptidoglycan (PG) by endogenous peptidoglycan hydrolases (PGHs) or autolysins. Origins of differences in autolytic properties of different strains are not fully elucidated. Regulation of autolysis possibly depends on the structure of the cell wall components other than PG, particularly polysaccharides. In the present work, we screened six L. helveticus strains with different autolytic properties: DPC4571, BROI and LH1. We established, for the first time, that cell walls (CWs) of these strains contained polysaccharides, different from their CW teichoic acids. Cell wall polysaccharides of three strains were purified, and their chemical structures were established by 2D NMR spectroscopy and methylation analysis. The structures of their repeating units are presented.


Measuring mining safety with injury statistics: lost workdays as indicators of risk.
Mining in the United States remains one of the most hazardous industries, despite significant reductions in fatal injury rates over the last century. Coal mine fatality rates, for example, have dropped almost a thousand-fold since their peak in 1908. While incidence rates are very important indicators, lost worktime measures offer an alternative metric for evaluating job safety and health performance. The first objective of this study examined the distributions and summary statistics of all injuries reported to the Mine Safety and Health Administration from 1983 through 2004. Over the period studied (1983-2004), there were 31,515,368 lost workdays associated with mining injuries, for an equivalent of 5,700 person-years lost annually. The second objective addressed the problem of comparing safety program performance in mines for situations where denominator data were lacking. By examining the consequences of injuries, comparisons can be made between disparate operations without the need for denominators. Total risk in the form of lost workday sums can help to distinguish between lower- and higher-risk operations or time periods. Our method was to use a beta distribution to model the losses and to compare underground coal mining to underground metal/nonmetal mining from 2000 to 2004. Our results showed the probability of an injury having 10 or more lost workdays was 0.52 for coal mine cases versus 0.35 for metal/nonmetal mine cases. In addition, a comparison of injuries involving continuous mining machines over 2001-2002 versus 2003-2004 showed that the ratio of average losses in the later period to those in the earlier period was approximately 1.08, suggesting increasing risks for such operations. This denominator-free safety measure will help the mining industry more effectively identify higher-risk operations and more realistically evaluate their safety improvement programs. Attention to a variety of metrics concerning the performance of a job safety and health program will enhance industry's ability to manage these programs and reduce risk.