Binding specificities of lectins to immobilized glycoproteins and oligosaccharides differ from those of immobilized lectins to oligosaccharides.
The carbohydrate-binding specificities of lectins in solution to glycoproteins and neoglycolipids immobilized on a solid phase were analyzed in order to establish a simple, rapid method for structural analysis of the carbohydrate moieties of small amounts of individual glycoproteins blotted on membrane. Eight glycoproteins containing typical O-linked tetrasaccharides or a series of typical N-linked oligosaccharides of the high-man-nose type, hybrid type, and complex type and 6 neoglycoproteins containing mono- or di-saccharides were dot blotted on membranes and the membranes were then reacted with 8 kinds of horseradish peroxidase-conjugated lectins before and after heat treatment. Neoglycolipids containing the glycoprotein-derived oligosaccharides immobilized on a thin layer chromatography plate were also reacted with lectins. The heat treatment of the membrane increased lectin reactivity toward the glycoproteins. The carbohydrate-binding behavior of lectins, Phaseolus vulgaris erythroagglutinin, wheat germ agglutinin, and concanavalin A in solution toward glycoproteins and neoglycolipids immobilized on a solid phase differed from that of immobilized lectins toward oligosaccharides in solution. This difference should be noted in lectin detection of specific carbohydrates of individual glycoproteins on membrane.


Depth-discrete metagenomics reveals the roles of microbes in biogeochemical cycling in the tropical freshwater Lake Tanganyika.
Lake Tanganyika (LT) is the largest tropical freshwater lake, and the largest body of anoxic freshwater on Earth's surface. LT's mixed oxygenated surface waters float atop a permanently anoxic layer and host rich animal biodiversity. However, little is known about microorganisms inhabiting LT's 1470 meter deep water column and their contributions to nutrient cycling, which affect ecosystem-level function and productivity. Here, we applied genome-resolved metagenomics and environmental analyses to link specific taxa to key biogeochemical processes across a vertical depth gradient in LT. We reconstructed 523 unique metagenome-assembled genomes (MAGs) from 34 bacterial and archaeal phyla, including many rarely observed in freshwater lakes. We identified sharp contrasts in community composition and metabolic potential with an abundance of typical freshwater taxa in oxygenated mixed upper layers, and Archaea and uncultured Candidate Phyla in deep anoxic waters. Genomic capacity for nitrogen and sulfur cycling was abundant in MAGs recovered from anoxic waters, highlighting microbial contributions to the productive surface layers via recycling of upwelled nutrients, and greenhouse gases such as nitrous oxide. Overall, our study provides a blueprint for incorporation of aquatic microbial genomics in the representation of tropical freshwater lakes, especially in the context of ongoing climate change, which is predicted to bring increased stratification and anoxia to freshwater lakes.


Should suspected cervical spinal cord injuries be immobilised? A systematic review protocol.
Cervical spinal cord injuries may result in life-threatening situations and long-term disability. Prehospital spinal immobilisation is the standard of care for patients with potential spinal cord injury (SCI). It aims to prepare patients for transport, achieve neutral spinal alignment, and reduce movement and secondary injuries in potentially unstable spines. However, there is a lack of evidence on its clinical benefits and its overall effect on patient outcomes. To identify the reported outcomes following immobilisation of suspected cervical SCI, to compare the effects of spinal immobilisation versus no immobilisation on the reported outcomes, and to provide recommendations for prehospital cervical immobilisation. A search of the literature will be conducted using relevant online databases. This will include all types of human studies that were published in English from the earliest record available to the first week of October 2013. One author will conduct the search and two independent authors will screen the titles and the abstracts identified by the search and critically appraise the selected papers. A third author will be available to resolve any disagreement. The findings will be reported according to Preferred Reporting Items for Systematic Reviews (PRISMA) guidelines. Critical appraisal as well as the level and the strength of evidence will follow the National Health and Medical Research Council (NHMRC) guidelines. Evidence-based practices should be pursued to further improve the prehospital care for suspected cervical SCI. This systematic review will contribute to the body of knowledge regarding the spinal immobilisation effects on the SCI patient's outcomes.


Role of 5p15.33 (TERT-CLPTM1L), 6p21.33 and 15q25.1 (CHRNA5-CHRNA3) variation and lung cancer risk in never-smokers.
Genome-wide association studies have provided evidence that common variation at 5p15.33 (TERT-CLPTM1L), 6p21.33 and 15q25.1 (CHRNA5-CHRNA3) influences lung cancer risk. To examine if variation at any of these loci influences the risk of lung cancer in never-smokers, we compared 5p15.33-TERT (rs2736100), 5p15.33-CLPTM1L (rs4975616), 6p21.33-BAT3 (rs3117582), 15q25.1-CHRNA3 (rs8042374) and 15q25.1-CHRNA3 (rs12914385) genotypes in a series of 239 never-smoker lung cancer cases and 553 never-smoker controls. A statistically significant association between lung cancer risk and 5p15.33 genotypes was found: rs2736100 (odds ratio = 0.78, 95% confidence interval: 0.63-0.97; P = 0.02), rs4975616 (odds ratio = 0.69, 95% confidence interval: 0.55-0.85; P = 7.95 x 10(-4)), primarily for adenocarcinoma. There was no evidence of association between 6p21.33 or 15q25.1 variation and risk of lung cancer. This analysis provides evidence that TERT-CLPTM1L variants may influence the risk of lung cancer outside the context of tobacco smoking.


Mammalian and avian embryology at Warsaw University (Poland) from XIX century to the present.
In this article, we describe the history (between the XIX century and World War II) of embryological research conducted at Warsaw University, together with current research activities being carried out at the Department of Embryology. During the partition of Poland, the Imperial (Russian) Warsaw University conducted research on avian embryology (and to a smaller extent, on reptilian embryology). When Poland regained independence in 1918, these studies were continued under the Chair of Comparative Anatomy headed by Professor Jan Tur. A new Department of Embryology created in 1954 was first headed by Professor Stanislaw Bilewicz and since 1964 by Professor Andrzej Tarkowski, who in 2003 was succeeded by Dr. Marek Maleszewski D.Sc. During the last 45 years, embryological research at Warsaw University has concentrated mainly on mammalian development with special emphasis on the regulative capabilities of early embryos and also on experimental chimaeras, nucleo-cytoplasmic interactions in oogenesis and early embryogenesis (including regulation of DNA replication and transcription), experimental parthenogenesis and fertilization.


Association of Ozone Exposure With Cardiorespiratory Pathophysiologic Mechanisms in Healthy Adults.
Exposure to ozone has been associated with cardiovascular mortality, but the underlying biological mechanisms are not yet understood. To examine the association between ozone exposure and cardiopulmonary pathophysiologic mechanisms. A longitudinal study involving 89 healthy adult participants living on a work campus in Changsha City, China, was conducted from December 1, 2014, to January 31, 2015. This unique quasiexperimental setting allowed for better characterization of air pollutant exposure effects because the participants spent most of their time in controlled indoor environments. Concentrations of indoor and outdoor ozone, along with the copollutants particulate matter, nitrogen dioxide, and sulfur dioxide, were monitored throughout the study period and then combined with time-activity information and filtration conditions of each residence and office to estimate 24-hour and 2-week combined indoor and outdoor mean exposure concentrations. Associations between each exposure measure and outcome measure were analyzed using single-pollutant and 2-pollutant linear mixed models controlling for ambient temperature, secondhand smoke exposure, and personal-level time-varying covariates. Biomarkers indicative of inflammation and oxidative stress, arterial stiffness, blood pressure, thrombotic factors, and spirometry were measured at 4 sessions. Of the 89 participants, 25 (28%) were women and the mean (SD) age was 31.5 (7.6) years. The 24-hour ozone exposure concentrations ranged from 1.4 to 19.4 parts per billion (ppb), corresponding to outdoor concentrations ranging from 4.3 to 47.9 ppb. Within this range, in models controlling for a second copollutant and other potential confounders, a 10-ppb increase in 24-hour ozone was associated with mean increases of 36.3% (95% CI, 29.9%-43.0%) in the level of platelet activation marker soluble P-selectin, 2.8% (95% CI, 0.6%-5.1%) in diastolic blood pressure, 18.1% (95% CI, 4.5%-33.5%) in pulmonary inflammation markers fractional exhaled nitric oxide, and 31.0% (95% CI, 0.2%-71.1%) in exhaled breath condensate nitrite and nitrate as well as a -9.5% (95% CI, -17.7% to -1.4%) decrease in arterial stiffness marker augmentation index. A 10-ppb increase in 2-week ozone was associated with increases of 61.1% (95% CI, 37.8%-88.2%) in soluble P-selectin level and 126.2% (95% CI, 12.1%-356.2%) in exhaled breath condensate nitrite and nitrate level. Other measured biomarkers, including spirometry, showed no significant associations with either 24-hour ozone or 2-week ozone exposures. Short-term ozone exposure at levels not associated with lung function changes was associated with platelet activation and blood pressure increases, suggesting a possible mechanism by which ozone may affect cardiovascular health.


[Renal tumors smaller that 3 cm].
To analyze the characteristics of renal adenocarcinoma less than 3 cm in size and to determine the therapeutic approach in these tumors. 286 cases with a histological diagnosis of renal adenocarcinoma that had undergone surgery at our hospital over the last 25 years were reviewed. The study showed 11 cases had renal tumors less than 3 cm in size. Six had been incidentally diagnosed. Two patients presented metastasis during the course of the disease and one patient had massive lymph node invasion (N3) at the time of diagnosis. Seven patients had a low tumor stage and grade. Renal tumors less than 3 cm in size have been considered to have a low potential for malignancy. However, the availability of more sophisticated diagnostic imaging techniques have led to earlier detection of renal masses, when the tumor is of a smaller size. Therefore determining whether the tumor is benign or malignant should be based exclusively on the histopathological findings. Treatment of this type of lesions is by surgery and performing a partial nephrectomy should be considered.


Joint learning framework of cross-modal synthesis and diagnosis for Alzheimer's disease by mining underlying shared modality information.
Alzheimer's disease (AD) is one of the most common neurodegenerative disorders presenting irreversible progression of cognitive impairment. How to identify AD as early as possible is critical for intervention with potential preventive measures. Among various neuroimaging modalities used to diagnose AD, functional positron emission tomography (PET) has higher sensitivity than structural magnetic resonance imaging (MRI), but it is also costlier and often not available in many hospitals. How to leverage massive unpaired unlabeled PET to improve the diagnosis performance of AD from MRI becomes rather important. To address this challenge, this paper proposes a novel joint learning framework of unsupervised cross-modal synthesis and AD diagnosis by mining underlying shared modality information, improving the AD diagnosis from MRI while synthesizing more discriminative PET images. We mine underlying shared modality information in two aspects: diversifying modality information through the cross-modal synthesis network and locating critical diagnosis-related patterns through the AD diagnosis network. First, to diversify the modality information, we propose a novel unsupervised cross-modal synthesis network, which implements the inter-conversion between 3D PET and MRI in a single model modulated by the AdaIN module. Second, to locate shared critical diagnosis-related patterns, we propose an interpretable diagnosis network based on fully 2D convolutions, which takes either 3D synthesized PET or original MRI as input. Extensive experimental results on the ADNI dataset show that our framework can synthesize more realistic images, outperform the state-of-the-art AD diagnosis methods, and have better generalization on external AIBL and NACC datasets.


Spatial model of Dengue Hemorrhagic Fever (DHF) risk: scoping review.
Creating a spatial model of dengue fever risk is challenging duet to many interrelated factors that could affect dengue. Therefore, it is crucial to understand how these critical factors interact and to create reliable predictive models that can be used to mitigate and control the spread of dengue. This scoping review aims to provide a comprehensive overview of the important predictors, and spatial modelling tools capable of producing Dengue Haemorrhagic Fever (DHF) risk maps. We conducted a methodical exploration utilizing diverse sources, i.e., PubMed, Scopus, Science Direct, and Google Scholar. The following data were extracted from articles published between January 2011 to August 2022: country, region, administrative level, type of scale, spatial model, dengue data use, and categories of predictors. Applying the eligibility criteria, 45 out of 1,349 articles were selected. A variety of models and techniques were used to identify DHF risk areas with an arrangement of various multiple-criteria decision-making, statistical, and machine learning technique. We found that there was no pattern of predictor use associated with particular approaches. Instead, a wide range of predictors was used to create the DHF risk maps. These predictors may include climatology factors (e.g., temperature, rainfall, humidity), epidemiological factors (population, demographics, socio-economic, previous DHF cases), environmental factors (land-use, elevation), and relevant factors. DHF risk spatial models are useful tools for detecting high-risk locations and driving proactive public health initiatives. Relying on geographical and environmental elements, these models ignored the impact of human behaviour and social dynamics. To improve the prediction accuracy, there is a need for a more comprehensive approach to understand DHF transmission dynamics.


Diversity versus concentration in antipsychotic drug selection.
It is unknown to what extent providers utilize the full range of available antipsychotic agents, or tend to prescribe just a few with which they are familiar. This study uses the Herfindahl index to measure the degree to which antipsychotic prescribing is diverse or concentrated within medical centers in the Department of Veterans Affairs. Patients with schizophrenia who received prescriptions for antipsychotic medications from the Department of Veterans Affairs in fiscal year 2005 were identified. For each facility, the proportion of prescriptions that were written for each antipsychotic medication was computed. The Herfindahl index, which is defined as the sum of the square of the proportionate shares of each medication (i.e., 50% squared = 0.25), was computed for each facility, and its distribution across facilities was examined. We identified 785,485 prescriptions for antipsychotic medications across 76,787 patients within 128 facilities. With 7 drugs to choose from (first-generation antipsychotics were not differentiated as separate drugs), the Herfindahl could range from 0.14 (if the drugs were prescribed equally) to 1 (if only 1 drug was prescribed to all patients). The Herfindahl averaged 0.20 to 0.21 and ranged from 0.16 to 0.31 across facilities. The Herfindahl index is a useful metric for describing the degree to which providers use a range of antipsychotic medications in treating patients with schizophrenia and suggests that VA facilities use a diversity of agents, thus taking advantage of the potentially unique drug-patient matches. The extent to which providers tailor their choice of antipsychotic medication to the particular characteristics of the patient may lead to better quality of care. The fact that providers do use a range of antipsychotic medications suggests that formulary policies should not reduce access to these drugs. Further research is needed to construct physician-level measures of prescribing diversity and to examine whether clinical outcomes are improved when providers use a wider range of medications in their practice.


Avian influenza monitoring in migrating birds in Taiwan during 1998-2007.
Avian influenza virus (AIV) monitoring in migratory birds has been performed in Taiwan since 1998. From 1998 to 2007, 29,287 samples were collected from wild ducks, shorebirds, and other wild birds in the four wetlands around Taiwan and at two outside islets, Penghu and Kinmen. Virus isolation was performed for all collected samples by inoculating chicken embryos. The AIV in the allantoic fluid was identified using hemagglutination and reverse transcription PCR. The AIV prevalence from those samples was 0.81% (237/29,287). The peak prevalence reached 1.06% (186/17,493) from September to December, during which time migrating ducks came from the North. The prevalence from January to April was 0.51%. However, no virus was isolated from May to August. The partial HA genes of 28 H4 AIVs were sequenced and analyzed. The phylogenetic tree showed that a correlation existed between the isolation years and the evolutional distances. The pathogenicity of the isolated H5 and H7 AIVs was determined by intravenous pathogenicity index (IVPI) testing in specific-pathogen-free chickens and by HA cleavage sequencing. Using the IVPI test and the HA cleavage sequences, all of the H5 or H7 AIVs isolated were determined to be low pathogenicity AIVs.


Induced cardiac pacemaker cells survive metabolic stress owing to their low metabolic demand.
Cardiac pacemaker cells of the sinoatrial node initiate each and every heartbeat. Compared with our understanding of the constituents of their electrical excitation, little is known about the metabolic underpinnings that drive the automaticity of pacemaker myocytes. This lack is largely owing to the scarcity of native cardiac pacemaker myocytes. Here, we take advantage of induced pacemaker myocytes generated by TBX18-mediated reprogramming (TBX18-iPMs) to investigate comparative differences in the metabolic program between pacemaker myocytes and working cardiomyocytes. TBX18-iPMs were more resistant to metabolic stresses, exhibiting higher cell viability upon oxidative stress. TBX18-induced pacemaker myocytes (iPMs) expensed a lower degree of oxidative phosphorylation and displayed a smaller capacity for glycolysis compared with control ventricular myocytes. Furthermore, the mitochondria were smaller in TBX18-iPMs than in the control. We reasoned that a shift in the balance between mitochondrial fusion and fission was responsible for the smaller mitochondria observed in TBX18-iPMs. We identified a mitochondrial inner membrane fusion protein, Opa1, as one of the key mediators of this process and demonstrated that the suppression of Opa1 expression increases the rate of synchronous automaticity in TBX18-iPMs. Taken together, our data demonstrate that TBX18-iPMs exhibit a low metabolic demand that matches their mitochondrial morphology and ability to withstand metabolic insult.


[Functional and ultrastructural changes in the adenohypophysis in obesity].
The indices of some tropic adenohypophyseal functions of obese patients were compared with ultrastructural reaction of the adequate adenocytes of rats with experimental hypothalamic obesity. It was shown that in obesity significant functional and structural changes occur in adenohypophyseal cells, their extent being considerably associated with the excessive body mass. In pronounced obesity, a decrease in somato- and thyrotropic- and an increase in gonado- and corticotropic adenohypophyseal functions are seen, manifesting in ultrastructural changes of appropriate adenocytes. The results obtained allow a conclusion, that one of the mechanisms of obesity development is the changed secretory potency of adenohypophyseal cells, producing hormones, which exert a substantial influence upon lipogenesis and lipolysis processes.


Data-Driven Sparse Skin Stimulation Can Convey Social Touch Information to Humans.
During social interactions, people use auditory, visual, and haptic cues to convey their thoughts, emotions, and intentions. Due to weight, energy, and other hardware constraints, it is difficult to create devices that completely capture the complexity of human touch. Here we explore whether a sparse representation of human touch is sufficient to convey social touch signals. To test this we collected a dataset of social touch interactions using a soft wearable pressure sensor array, developed an algorithm to map recorded data to an array of actuators, then applied our algorithm to create signals that drive an array of normal indentation actuators placed on the arm. Using this wearable, low-resolution, low-force device, we find that users are able to distinguish the intended social meaning, and compare performance to results based on direct human touch. As online communication becomes more prevalent, such systems to convey haptic signals could allow for improved distant socializing and empathetic remote human-human interaction.


Pedunculated inverted hyperplastic polyp of the sigmoid colon treated with endoscopic polypectomy.
A case of inverted hyperplastic polyp of the sigmoid colon is reported. The patient was a 67-year-old woman who visited our hospital for further evaluation of constipation. Colonoscopy revealed a pedunculated polyp with linear central depression, about 12 mm in diameter, in the sigmoid colon. Excluding the polyp, there was no lesion in the colorectum. Endoscopic polypectomy was performed. Histological examination of the specimen revealed a stalked polyp that had scattered goblet cells and elongated tubules with serrated profiles in the superficial portion. The nodules of deep tubules impinged on the muscularis mucosae. Proliferation of fibromuscular tissue was not seen in the polyp. There was no evidence of malignancy. This polyp was diagnosed as an inverted hyperplastic polyp. Pedunculated-type inverted hyperplastic polyp of the colon is rare.


The "Nouvelle Vague" of surgeons in pancreatic surgery: can they rise to the legacy of current surgeons?
Pancreatic surgery is still thought as a challenging field even for experienced hepatobilliary (HPB) surgeons and high volume tertiary centers. The purpose of this study was to present the results (mortality and morbidity) of pancreatic surgery in a high volume center, in operations performed solely by inexperienced surgeons (two 6th year residents and a HPB fellow) under the supervision of expert surgeons on the field. Forty-one consecutive patients who underwent curative-intent pancreatic resection with a modified pancreaticojejunostomy between January 2010 and December 2014 at Asklepios Hospital Barmbek, Germany, were identified from our institutional computer-based database. Two 6th year residents and an HPB-fellow performed all pancreatic anastomoses under the instructions of an experienced surgeon. Perioperative outcomes were recorded and analyzed. Median postoperative length of stay for all patients was 15 days (IQR:7-31). In the first 90 postoperative days, the postoperative mortality rate was 0% and morbidity rate reached 39%. Reoperation was required in 1 patient (2.44%). However, no reoperation was performed for pancreatic anastomotic failure. No postoperative hemorrhage requiring interventional procedure or reoperation occurred in any patient. The outcomes of pancreatic surgery performed by less experienced surgeons are satisfactory. The instructions of an expert surgeon in a high volume hospital definitely secures a favorable outcome after pancreatic surgery with lower mortality and morbidity rates compared with current literature trends.


Precision proteomics: the case for high resolution and high mass accuracy.
Proteomics has progressed radically in the last 5 years and is now on par with most genomic technologies in throughput and comprehensiveness. Analyzing peptide mixtures by liquid chromatography coupled to high-resolution mass spectrometry (LC-MS) has emerged as the main technology for in-depth proteome analysis whereas two-dimensional gel electrophoresis, low-resolution MALDI, and protein arrays are playing niche roles. MS-based proteomics is rapidly becoming quantitative through both label-free and stable isotope labeling technologies. The latest generation of mass spectrometers combines extremely high resolving power, mass accuracy, and very high sequencing speed in routine proteomic applications. Peptide fragmentation is mostly performed in low-resolution but very sensitive and fast linear ion traps. However, alternative fragmentation methods and high-resolution fragment analysis are becoming much more practical. Recent advances in computational proteomics are removing the data analysis bottleneck. Thus, in a few specialized laboratories, "precision proteomics" can now identify and quantify almost all fragmented peptide peaks. Huge challenges and opportunities remain in technology development for proteomics; thus, this is not "the beginning of the end" but surely "the end of the beginning."


Association of emergency allotment discontinuation with household food insufficiency in Supplemental Nutrition Assistance Program participants: A quasi-experimental study.
The Supplemental Nutrition Assistance Program (SNAP) was modified to mitigate food insecurity faced by low-income families during the pandemic. These changes included enhancement of SNAP benefits via 2020 emergency allotments (EA). Despite the high food price inflation in 2022, 17 states ceased providing EA benefits by end of 2022. The objective of this research is to examine the impact of EA discontinuation on food insufficiency. Using Household Pulse Survey data from December 2021-January 2023, SNAP participants (n = 57,556) from states that discontinued EA at some point during the study period and those from states that did not discontinue EA were included. The main analyses examined the impact of EA discontinuation on food insufficiency via staggered difference-in-difference models using two-way fixed effects (TWFE) regression. Additional analyses examined this association using Callaway-Sant'Anna approach for difference-in-difference analyses. Using TWFE, EA discontinuation was significantly associated with increased food insufficiency in the overall sample, as well as in individuals who were non-Hispanic White and Hispanic, and who had annual family incomes of less than $25,000. Investigation of time-varying association of EA termination with food insufficiency suggested that EA discontinuation was significantly associated with greater food insufficiency in the second week following rollback. DiD analyses using Callaway-Sant'Anna approach suggested that states' rollback of EA was significantly associated with higher food insufficiency in non-Hispanic White individuals. Findings imply that EA discontinuation, amidst the corresponding surge in inflation, could have contributed to increased rates of household food insufficiency.


Studies of phagocytosis of group A streptococci by polymorphonuclear leucocytes in vitro.
Studies have been made on phagocytosis and killing of Group A streptococci during mixing with suspensions of leucocytes in vitro. Under appropriate test conditions an anti-phagocytic effect can be demonstrated for the streptococcal hyaluronic acid capsule as well as for its M protein. The results obtained suggest an explanation for the suitability of human, but not rabbit, blood for opsonophagocytic tests designed to measure type-specific streptococcal antibodies. Human sera contain a factor which counteracts the anti-phagocytic effects of streptococcal hyaluronic acid capsules, and hence human blood serves well for detection of antibodies which combine with the only other phagocytosis-resisting component of this microorganism, namely M protein. In contrast, rabbit sera contain none of this factor, and addition of antibody to M protein to phagocytic test systems employing rabbit serum does not necessarily render the streptococci susceptible to engulfment by white cells, since the hyaluronic acid capsule may continue to interfere with phagocytosis. The nature of the human serum factor which opsonizes encapsulated streptococci is unknown. It does not appear to be an antibody or an enzyme capable of depolymerizing hyaluronic acid.


Quantification of polymerase chain reaction products by affinity-based hybrid collection.
We have used oligonucleotides modified with biotin in the 5'-end as primers in the polymerase chain reaction (PCR)-amplification. This results in the synthesis of 5'-biotinylated DNA molecules, which are detected by hybridization to a labelled probe in solution. The formed hybrids are collected on an avidin-matrix by mediation of the biotin residue of the target molecules. The affinity-based hybrid collection method is quantitative and makes it possible to measure the amount of DNA produced in the PCR-amplification. At low concentrations of template the efficiency of the process is close to 100%, making it possible to detect the presence of a few molecules of target DNA in 25 cycles. With high template concentrations the efficiency of the process is low.


Assessing the impact of policy changes in the Icelandic cod fishery using a hybrid simulation model.
Most of the Icelandic cod is caught in bottom trawlers or longliners. These two fishing methods are fundamentally different and have different economic, environmental, and even social effects. In this paper we present a hybrid-simulation framework to assess the impact of changing the ratio between cod quota allocated to vessels with longlines and vessels with bottom trawls. It makes use of conventional bioeconomic models and discrete event modelling and provides a framework for simulating life cycle assessment (LCA) for a cod fishery. The model consists of two submodels, a system dynamics model describing the biological aspect of the fishery and a discrete event model for fishing activities. The model was run multiple times for different quota allocation scenarios and results are presented where different scenarios are presented in the three dimensions of sustainability: environmental, social, and economic. The optimal allocation strategy depends on weighing the three different factors. The results were encouraging first-steps towards a useful modelling method but the study would benefit greatly from better data on fishing activities.


Reducing major cause-specific hospitalization rates and shortening hospital stays after influenza vaccination.
The impact of influenza vaccination on major cause-specific hospitalization and the duration of hospital stay is rarely reported. Our purpose was to study the effect of vaccine efficacy on major disease-specific hospitalization and the duration of hospital stays among elderly persons. From 1 January through 30 June 2001, we prospectively observed 35,637 vaccinated elderly persons (age, >or=65 years) and 53,094 unvaccinated elderly persons in Kaohsiung County, Taiwan, by computerized linkage to the National Health Insurance database. Of these persons, 21,347 had been assigned a high-risk status by the Department of Health, Taiwan. Univariate and multivariate logistic regression were used for determining vaccine efficacy in hospitalization. Multiple linear regression analyses were performed for determining the length of hospital stays. In both high-risk and low-risk groups, vaccination was associated with reducing the rates of hospitalization for all causes (20% vs. 23%), lung diseases, congestive heart failure (43% vs. 32%), renal disease, and liver disease (P<.05). It was also significant for stroke, hypertension, diabetes, neoplasm, and injury in low-risk patients (P<.05). Multivariate logistic regression showed that vaccination was significantly associated with reducing the rate of hospitalization (odds ratio [OR], 0.89; 95% confidence interval [CI], 0.86-0.92), but those with high-risk status had an increased risk of hospitalization (OR, 3.69; 95% CI, 3.56-3.82). Multiple linear regression analysis showed that vaccination decreased the duration of all-cause hospital stays (coefficient, -2.4 days; 95% CI, -2.7 to -2.1 days) and of hospitalization due to lung disease (coefficient, -4.9 days; 95% CI, -6.0 to -3.8 days). Influenza vaccination may reduce hospitalization rates and shorten hospital stays not only for lung diseases but also for other common diseases in high-risk and low-risk elderly populations.


[Efficacy and safety in the new regulation of pharmaceutical products in Peru].
Internationally, pharmaceutical products (FP) are authorized after their risk-benefit profile has been assessed, taking into account efficacy, safety and quality parameters. In 2009, it was established in Peru that all pharmaceutical products to be commercialized should show proof of their efficacy and safety. According to the new regulation, in effect as of 2012, the pharmaceutical specialties (FS) have been grouped into three categories: category 1, if included in the National List of Essential Medicines; category 2, if authorized in countries with high health surveillance; and category 3, if not included in categories 1 or 2. The scientific documentation to be submitted for the registration or re-registration of the product in the regulatory entity will depend on its FS category.


Effects of alcohol and extended practice on divided-attention performance.
The effects of alcohol and extended practice on divided attention were investigated using a visual tracking task and an auditory detection task. Subjects performed the tasks with and without alcohol, under single-task (S) and dual-task (D) conditions, both before and after they had received extended practice under single-task conditions without alcohol. Tracking accuracy improved with practice and was impaired under divided-attention conditions but was not affected by alcohol. Speed of detection was impaired by alcohol, improved by practice, and impaired by divided attention. Extended practice did not reduce the influence of alcohol. The effects of both alcohol and practice on speed of detection were significantly greater under dual-task conditions than under single-task conditions. Analysis of detection-task reaction times in terms of relative divided-attention costs, (D-S)/S, showed no effect of alcohol, but a highly significant reduction in costs with extended practice. It is concluded that (1) alcohol and practice can have quantitatively, but not qualitatively, similar effects on speeded performance, and (2) the effect of alcohol is not influenced by the attentional requirements of the task.


Quality indicators for depression care in HIV patients.
HIV patients are at increased risk for depression. However, a comprehensive set of quality of care indicators for depression in the context of HIV does not exist. We report the results of a recent expert panel convened to develop a comprehensive set of HIV depression quality indicators. We systematically reviewed the depression and HIV depression literature to provide the evidence for the quality indicators. The HIV depression quality indicators were divided into six chapters: general, bereavement, substance abuse, viral hepatitis, cognitive impairment and drug interactions. A modified Delphi expert panel method was used to evaluate the validity and feasibility of the candidate quality indicators. The expert panel included HIV physicians, psychiatrists and a pharmacist, all with experience with HIV depression. The in-person eleven-member panel rated 80 candidate HIV-depression quality indicators. Indicators were evaluated using main and sensitivity analysis criteria for validity and feasibility. Seventy-four percent (59/80) of the quality indicators met a priori main analysis criteria for validity and feasibility and 61% met sensitivity analysis criteria. Approved indicators that are more specific to HIV depression include: depression screening frequency; depression screening and treatment in the context of hepatitis C; optimizing depression and HIV treatment in the context of cognitive impairment; and starting antidepressants at lower doses but advancing to doses typical for the general population. Most adverse medication interaction indicators did not meet main analysis validity criteria. The quality indicators identified in this study provide a useful tool for measuring and informing the quality of HIV-depression care.


A practical algorithm for reconstructing level-1 phylogenetic networks.
Recently, much attention has been devoted to the construction of phylogenetic networks which generalize phylogenetic trees in order to accommodate complex evolutionary processes. Here, we present an efficient, practical algorithm for reconstructing level-1 phylogenetic networks--a type of network slightly more general than a phylogenetic tree--from triplets. Our algorithm has been made publicly available as the program LEV1ATHAN. It combines ideas from several known theoretical algorithms for phylogenetic tree and network reconstruction with two novel subroutines. Namely, an exponential-time exact and a greedy algorithm both of which are of independent theoretical interest. Most importantly, LEV1ATHAN runs in polynomial time and always constructs a level-1 network. If the data are consistent with a phylogenetic tree, then the algorithm constructs such a tree. Moreover, if the input triplet set is dense and, in addition, is fully consistent with some level-1 network, it will find such a network. The potential of LEV1ATHAN is explored by means of an extensive simulation study and a biological data set. One of our conclusions is that LEV1ATHAN is able to construct networks consistent with a high percentage of input triplets, even when these input triplets are affected by a low to moderate level of noise.


Methods to Succeed in Effective Knowledge Translation in Clinical Practice.
To explore the evidence around facilitation as an intervention for the successful implementation of new knowledge into clinical practice. The revised version of the Promoting Action on Research Implementation in Health Services (PARIHS) framework, called the integrated or i-PARIHS framework, is used as the explanatory framework. This framework posits that evidence is a multidimensional construct embedded within innovation and operationalized by clinicians (individuals and within teams), working across multiple layers of context. Facilitation is the active ingredient that promotes successful implementation. An emerging body of evidence supports facilitation as a mechanism to getting new knowledge into clinical practice. Facilitation roles are divided into beginner, experienced, and expert facilitators. Facilitators can be internal or external to the organization they work in, and their skills and attributes complement other knowledge translation (KT) roles. Complex KT projects require facilitators who are experienced in implementation methods. Facilitation is positioned as the active ingredient to effectively introduce new knowledge into a clinical setting. Levels of facilitation experience are assessed in relation to the complexity of the KT task. Three core facilitation roles are identified, and structured interventions are established taking into account the nature and novelty of the evidence, the receptiveness of the clinicians, and the context or setting where the new evidence is to be introduced. Roles such as novice, experienced, and expert facilitators have important and complementary parts to play in enabling the successful translation of evidence into everyday practice in order to provide effective care for patients.


Molecular Mechanisms of Colistin Resistance in Klebsiella pneumoniae Causing Bacteremia from India-A First Report.
Colistin has long been a reserve drug used for the treatment of carbapenem resistant Klebsiella pneumoniae. Carbapenem resistance in K. pneumoniae has been increasing and is as high as 44% in India. Although a reserve agent, with rise in rates of resistance to carbapenems, the usage of colistin has increased over the years leading to slow emergence of resistance. Colistin resistance is mainly mediated by the alteration in the LPS of bacterial outer membrane with the addition of L-Ara4-N and PEtN molecules. These alterations are mediated by mutations in several genes involved in lipidA modifications and most commonly mutations in mgrB gene has been reported. Recently there is emergence of plasmid mediated resistance due to mcr-1 and mcr-2 genes which poses a threat for the rapid global spread. This study aims at characterizing eight colistin resistant K. pneumoniae from bacteremia by whole genome sequencing. Eight K. pneumoniae were isolated from blood culture during 2013 and 2014 at the Department of Clinical Microbiology, Christian Medical College, India. Antimicrobial susceptibility testing was performed and minimum inhibitory concentration (MIC) was determined for colistin and polymyxin B by broth-micro dilution method. Whole genome sequencing was performed using Ion Torrent and the genome of all eight isolates was analyzed. The eight isolates were resistant to all the antimicrobials expect tigecycline. MIC of colistin and polymyxin B were ranged from 4 to 1024 μg/ml and 0.5 to 2048 μg/ml respectively. Multiple mutations were observed in the chromosomal genes involved in lipid A modifications. mcr-1 and mcr-2 gene was absent in all the isolates. The most significant were mutations in mgrB gene. Among the eight isolates, four, three and one were belonged to sequence types ST 231, ST14 and ST147 respectively. Seven isolates had blaOXA-48 like, one co-expressed blaNDM-1 and blaOXA-48 like genes leading to carbapenem resistance. Overall, multiple numbers of alterations have been observed. This includes silent mutations, point mutations, insertions and/or deletions. Mutations in mgrB gene is responsible for resistance to colistin in this study. Due to emergence of resistance to reserve drugs, there is a need for combination therapies for carbapenem resistant K. pneumoniae and colistin must be judiciously used.


Endodontic and aesthetic management of a conoid tooth with type IIIA dens invaginatus.
Dens invaginatus is a dental developmental abnormality caused by an infolding of the enamel organ into the adjacent dental papilla before the calcification of the dental tissues. The clinical presentation of dens invaginatus varies according to its severity, from mild forms, in which we might find a deeper than normal cingulum pit, to severe cases, in which there is a pronounced infolding reaching the apical foramen. The very nature of the invagination favours the contamination from the oral environment to the pulp space and the onset of caries. Therefore, patients might present with symptoms or signs of pulpitis and/or apical periodontitis. The aim of this paper is to present the successful endodontic and restorative management of a conoid shaped upper lateral incisor, affected by dens invaginatus type IIIA. A 13-year-old girl was referred to our office for endodontic and aesthetic treatment of the right upper lateral incisor, which was asymptomatic. Clinical, radiographic and photographic examinations were performed. The diagnosis of this tooth was dens invaginatus, Oehlers type IIIA with chronic apical periodontitis. Full isolation and magnification were used to achieve correct endodontic management of the aberrant internal anatomy of the tooth. Ultrasonic tips and a combination of manual and mechanical instruments were used for cleaning and shaping. Warm vertical condensation technique was used for sealing the canal system. As the final restoration it was chosen an indirect composite crown bonded to the tooth surface with minor preparation and using an adhesive technique. Radiographic healing and satisfaction of the aesthetic needs was achieved up to 5 years from completion of treatment. This case report shows the successful outcomes of a complex case based on proper diagnosis and correct treatment strategies for arresting the progression of apical periodontitis in challenging internal anatomies, as well as the fulfillment of a young patient aesthetic needs.


Molecular epidemiology and clinical spectrum of hereditary spastic paraplegia in the Japanese population based on comprehensive mutational analyses.
Hereditary spastic paraplegia (HSP) is one of the most genetically heterogeneous neurodegenerative disorders characterized by progressive spasticity and pyramidal weakness of lower limbs. Because >30 causative genes have been identified, screening of multiple genes is required for establishing molecular diagnosis of individual patients with HSP. To elucidate molecular epidemiology of HSP in the Japanese population, we have conducted mutational analyses of 16 causative genes of HSP (L1CAM, PLP1, ATL1, SPAST, CYP7B1, NIPA1, SPG7, KIAA0196, KIF5A, HSPD1, BSCL2, SPG11, SPG20, SPG21, REEP1 and ZFYVE27) using resequencing microarrays, array-based comparative genomic hybridization and Sanger sequencing. The mutational analysis of 129 Japanese patients revealed 49 mutations in 46 patients, 32 of which were novel. Molecular diagnosis was accomplished for 67.3% (33/49) of autosomal dominant HSP patients. Even among sporadic HSP patients, mutations were identified in 11.1% (7/63) of them. The present study elucidated the molecular epidemiology of HSP in the Japanese population and further broadened the mutational and clinical spectra of HSP.


Enhanced efficacy with reduced toxicity of chemotherapy drug 5-fluorouracil by synergistic treatment with Abnormal Savda Munziq from Uyghur medicine.
Abnormal Savda Munziq (ASMq) is a traditional prescription in Uyghur Medicine, and its treatment of complex diseases such as tumors and asthma has been proven to be effective in Uyghur medical clinical practice. The efficacy-enhancing and toxicity-reducing properties of ASMq were studied on mice with transplanted cervical cancer (U27) tumors, which were treated with 5-fluorouracil (5-FU) in this work. To investigate the synergistic effect of ASMq and 5-FU on U27 cells, inhibitory effects on cell proliferation were determined through a MTT assay. 48 Kunming mice which were randomly divided in to 6 groups: control group, model group, 5-FU group, 5-FU combine with ASMq low-dose group, 5-FU combine with ASMq medium-dose group, and 5-FU combine with ASMq high- dose group, the inhibition rate of the tumor, the viscera indexes, and the content of serum tumor necrosis factor-α (TNF-α), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were determined. The expression levels of transforming growth factor-β1 (TGF-β1) and human papillomavirus type 16 E2 (HPV16 E2) protein were assessed by Western blot. Pathological changes in the liver were observed. The inhibition rates of tumors, the 5-FU + ASMq.H group(80.64%), 5-FU + ASMq.M group (90.67%), 5-FU + ASMq.L group (72.03%) and 5-FU group (66.89%), clearly indicated that the effects of tumor inhibition. The thymus index and spleen index were increased, and the serum concentration of TNF-α increased while ALT and AST concentrations were decreased, and TNF-α protein expression were increased while TGF-β1 and HPV16 E2 were decreased. ASMq might can improve livers central vein hyperemia and interstitial edema, and preserve the radial structure of the hepatic cords. The results suggested that ASMq might reduce toxicity and enhance the efficacy of the chemotherapeutic drug 5-fluorouracil in the treatment of cervical carcinoma.


Immunohistochemistry of p63 in primary and secondary vulvar Paget's disease.
Vulvar Paget's disease (VPD) is classified into primary and secondary types. Differentiation of these subsets in biopsy specimen is important for appropriate therapy. Expression profile of cytokeratin (CK) 7 and CK20, gross cystic disease fluid protein-15 and uroplakin III has been reported as a differentiation marker of primary and secondary VPD. To examine the role of p63 immunostaining in differential diagnosis between primary VPD and VPD secondary to urothelial carcinoma (VPD-UC), expression of p63 was examined in nine cases of VPD. Paget cells in seven cases of VPD without UC did not express p63. In two cases of VPD associated with UC, Paget cells and UC cells had identical CK expression profile. UC cells were positive for p63 in both cases. In one case, Paget cells were positive for p63 and examination of the resected specimen showed that VPD was secondary to UC. In another case, Paget cells were negative for p63 and this was diagnosed as primary VPD independent of UC. This indicates that p63 is absent in Paget cells in primary VPD and is therefore useful in differentiating primary VPD from VPD-UC.


Field studies on the environmental fate of the Cry1Ab Bt-toxin produced by transgenic maize (MON810) and its effect on bacterial communities in the maize rhizosphere.
Field studies were done to assess how much of the transgenic, insecticidal protein, Cry1Ab, encoded by a truncated cry1Ab gene from Bacillus thuringiensis (Bt), was released from Bt-maize MON810 into soil and whether bacterial communities inhabiting the rhizosphere of MON810 maize were different from those of the rhizosphere of nontransgenic maize cultivars. Bacterial community structure was investigated by SSCP (single-strand conformation polymorphism) of PCR-amplified 16S rRNA genes from community DNA. Using an improved extraction and detection protocol based on a commercially available ELISA, it was possible to detect Cry1Ab protein extracted from soils to a threshold concentration of 0.07 ng/g soil. From 100 ng of purified Cry1Ab protein added per gram of soil, only an average of 37% was extractable. At both field sites investigated, the amount of Cry1Ab protein in bulk soil of MON810 field plots was always lower than in the rhizosphere, the latter ranging from 0.1 to 10 ng/g soil. Immunoreactive Cry1Ab protein was also detected at 0.21 ng/g bulk soil 7 months after harvesting, i.e. in April of the following year. At this time, however, higher values were found in residues of leaves (21 ng/g) and of roots (183 ng/g), the latter corresponding to 12% of the Cry1Ab protein present in intact roots. A sampling 2 months later indicated further degradation of the protein. Despite the detection of Cry1Ab protein in the rhizosphere of MON810 maize, the bacterial community structure was less affected by the Cry1Ab protein than by other environmental factors, i.e. the age of the plants or field heterogeneities. The persistence of Cry1Ab protein emphasizes the importance of considering post-harvest effects on nontarget organisms.


Routine abdominal drainage for uncomplicated liver resection.
The main reasons for inserting a drain after elective liver resections are (i) prevention of sub-phrenic or sub-hepatic fluid collection; (ii) identification and monitoring of post-operative bleeding; (iii) identification and drainage of any bile leak; and (iv) prevent the accumulation of ascitic fluid in cirrhotics. However, there are reports that drain use increases the complication rates. To assess the benefits and harms of routine abdominal drainage in elective liver resections. We searched The Cochrane Hepato-Biliary Group Controlled Trials Register, the Cochrane Central Register of Controlled Trials (CENTRAL) in The Cochrane Library, MEDLINE, EMBASE, and Science Citation Index Expanded until March 2007. We included all randomised trials comparing abdominal drainage and no drainage in adults undergoing elective liver resection. We also included randomised trials comparing different types of drain in adults undergoing elective liver resection. We collected the data on the characteristics of the trial, methodological quality of the trials, mortality, morbidity, conversion rate, operating time, and hospital stay from each trial. We analysed the data with both the fixed-effect and the random-effects models using the Cochrane Collaboration statistical software RevMan Analysis. For each outcome we calculated the odds ratio (OR) with 95% confidence intervals (CI) (based on intention-to-treat analysis) by combining the trial data sets using fixed-effect model or random-effects model, as appropriate. Drain versus no drain: We included five trials with 465 patients randomised: 234 to the drain group and 231 to the no drain group. Three of the five trials were of high methodological quality. There was no statistically significant difference between the two groups for any of the outcomes (mortality, intra-abdominal collections requiring re-operation, infected intra-abdominal collections, wound infection, ascitic leak, and hospital stay, when the random-effects model was adopted. Open drain versus closed drain: One randomised clinical trial of low methodological quality comparing open with closed drainage (186 patients) showed a lower incidence of infected intra-abdominal collections, chest complications, and hospital stay in the closed drain group. There is no evidence to support routine drain use after uncomplicated liver resections.


AtRabF2b (Ara7) acts on the vacuolar trafficking pathway in tobacco leaf epidermal cells.
Rab GTPases are universal key regulators of intracellular secretory trafficking events. In particular, Rab 5 homologues have been implicated in endocytic events and in the vacuolar pathway. In this study, we investigate the location and function of a member of this family, AtRabF2b (Ara7) in tobacco (Nicotiana tabacum) leaf epidermal cells using a live cell imaging approach. Fluorescent-tagged AtRabF2b[wt] localized to the prevacuolar compartment and Golgi apparatus, as determined by coexpression studies with fluorescent markers for these compartments. Mutations that impair AtRabF2b function also alter the subcellular location of the GTPase. In addition, coexpression studies of the protein with the vacuole-targeted aleurain-green fluorescent protein (GFP) and rescue experiments with wild-type AtRabF2b indicate that the dominant-negative mutant of AtRabF2b causes the vacuolar marker to be secreted to the apoplast. Our results indicate a clear role of AtRabF2b in the vacuolar trafficking pathway.


The role of motor intention in motor awareness: an experimental study on anosognosia for hemiplegia.
Recent theories propose that anosognosia for hemiplegia (AHP) results from specific impairments in motor planning. However, no study has hitherto directly investigated the role of motor intention in the observed non-veridical awareness of action in AHP. We developed the following paradigm to investigate the role of motor planning in awareness in patients with AHP: Four hemiplegic patients with and four without anosognosia were provided with false visual feedback of movement in their left paralysed arm through a prosthetic rubber hand. We examined whether the ability to detect presence or absence of movement based on visual evidence varied according to whether the patient had planned to move their limb or not. Motor intention had a selective effect on patients with AHP; they were more likely than controls (U = 16, P < 0.001) to ignore the visual feedback of a motionless hand and claim that they moved it when they had the intention to do so (self-generated movement) than when they expected an experimenter to move their own hand (externally generated movement), or there was no expectation of movement. By contrast, patients without AHP were not influenced by these manipulations, and did not claim they moved their hand when the hand remained still. This is the first direct demonstration that altered awareness of action in AHP reflects a dominance of motor intention prior to action over sensory information about the actual effects of movement.


Oil droplet ingestion and oil fouling in the copepod Calanus finmarchicus exposed to mechanically and chemically dispersed crude oil.
The rates of ingestion of oil microdroplets and oil fouling were investigated in the zooplankton filter-feeder Calanus finmarchicus (Gunnerus, 1770) at 3 concentrations of oil dispersions ranging from 0.25 mg/L to 5.6 mg/L. To compare responses to mechanically and chemically dispersed oil, the copepods were exposed to comparable dispersions of micron-sized oil droplets made with and without the use of a chemical dispersant (similar oil droplet size range and oil concentrations) together with a constant supply of microalgae for a period of 4 d. The filtration rates as well as accumulation of oil droplets decreased with increasing exposure concentration. Thus the estimated total amount of oil associated with the copepod biomass for the 2 lowest exposures in the range 11 mL/kg to 17 mL/kg was significantly higher than the approximately 6 mL/kg found in the highest exposure. For the 2 lowest concentrations the filtration rates were significantly higher in the presence of chemical dispersant. Furthermore, a significant increase in the amount of accumulated oil in the presence of dispersant was observed in the low exposure group.


Expression patterns of aurora kinase B, heat shock protein 47, and periostin in esophageal squamous cell carcinoma.
To elucidate the proteins related to the development and progress of esophageal squamous cell carcinoma (ESCC), we performed gene expression profile analysis of 14 ESCC tissues. We identified 182 genes that were commonly upregulated (p < 10(-5)) and 54 genes that were downregulated in ESCC tissues (p < 10(-6)). In order to validate the gene expression profiles, we did semiquantitative RT-PCR analysis and found 11 genes upregulated in greater than 70% of 19 tissue samples. We further examined the protein expression level and the distribution patterns of four genes: aurora kinase B (AURKB), periostin (POSTN), heat shock protein 47 (HSP47), and matrix metalloprotease 1 (MMP1). Ultimately, three genes (AURKB, HSP47, POSTN) were verified to be increased at both the mRNA and protein levels. Among them, AURKB and HSP47 were found to increase in the early development stage of ESCC and POSTN might be considered to function in the cell-cell interactions between cancer cell and adjacent stromal cells. Accordingly, these studies may provide valuable information for the identification of the genes that are necessary to study further their functions. Moreover, these genes might be used as potential diagnostic or therapeutic target molecules for ESCC patients.


Assessing cortical bone mechanical properties using collagen proton fraction from ultrashort echo time magnetization transfer (UTE-MT) MRI modeling.
Cortical bone shows as a signal void when using conventional clinical magnetic resonance imaging (MRI). Ultrashort echo time MRI (UTE-MRI) can acquire high signal from cortical bone, thus enabling quantitative assessments. Magnetization transfer (MT) imaging combined with UTE-MRI can indirectly assess protons in the organic matrix of bone. This study aimed to examine UTE-MT MRI techniques to estimate the mechanical properties of cortical bone. A total of 156 rectangular human cortical bone strips were harvested from the tibial and femoral midshafts of 43 donors (62 ± 22 years old, 62 specimens from females, 94 specimens from males). Bone specimens were scanned using UTE-MT sequences on a clinical 3 T MRI scanner and on a micro-computed tomography (μCT) scanner. A series of MT pulse saturation powers (400°, 600°, 800°) and frequency offsets (2, 5, 10, 20, 50 kHz) was used to measure the macromolecular fraction (MMF) utilizing a two-pool MT model. Failure mechanical properties of the bone specimens were measured using 4-point bending tests. MMF from MRI results showed significant strong correlations with cortical bone porosity (R = -0.72, P < 0.01) and bone mineral density (BMD) (R = +0.71, P < 0.01). MMF demonstrated significant moderate correlations with Young modulus, yield stress, and ultimate stress (R = 0.60-0.61, P < 0.01). These results suggest that the two-pool UTE-MT model focusing on the organic matrix of bone can potentially serve as a novel tool to detect the variations of bone mechanical properties and intracortical porosity.


What therapies are on the horizon for HER2 positive breast cancer?
Introduction: Despite dramatic improvements in survival achieved with currently available anti-HER2 agents, HER2-positive metastatic breast cancer remains an almost invariably deadly disease, with primary or acquired resistance to HER2-directed agents developing during treatment. Many efforts are focused on identifying new agents that may more effectively inhibit HER2 signaling and on possible combination strategies. Areas covered: This review summarizes the landscape of drugs under development for HER2-positive metastatic breast cancer, as antibody-drug conjugates, monoclonal anti-HER2 antibodies, bispecific antibodies, or novel tyrosine kinase inhibitors. Moreover, available data for possible combination of anti-HER2 drugs and different agents, as immunotherapy, PI3K/mTOR inhibitors, CDK4/6 inhibitors currently under evaluation are reviewed. These strategies may overcome mechanisms of resistance and further improve patient outcomes. Expert opinion: Identification of valuable predictive biomarkers is needed to better inform choice of treatment sequence for the individual patient and limit the financial toxicity of these agents.


Disruption of the A-kinase anchoring domain in flagellar radial spoke protein 3 results in unregulated axonemal cAMP-dependent protein kinase activity and abnormal flagellar motility.
Biochemical studies of Chlamydomonas flagellar axonemes revealed that radial spoke protein (RSP) 3 is an A-kinase anchoring protein (AKAP). To determine the physiological role of PKA anchoring in the axoneme, an RSP3 mutant, pf14, was transformed with an RSP3 gene containing a mutation in the PKA-binding domain. Analysis of several independent transformants revealed that the transformed cells exhibit an unusual phenotype: a fraction of the cells swim normally; the remainder of the cells twitch feebly or are paralyzed. The abnormal/paralyzed motility is not due to an obvious deficiency of radial spoke assembly, and the phenotype cosegregates with the mutant RSP3. We postulated that paralysis was due to failure in targeting and regulation of axonemal cAMP-dependent protein kinase (PKA). To test this, reactivation experiments of demembranated cells were performed in the absence or presence of PKA inhibitors. Importantly, motility in reactivated cell models mimicked the live cell phenotype with nearly equal fractions of motile and paralyzed cells. PKA inhibitors resulted in a twofold increase in the number of motile cells, rescuing paralysis. These results confirm that flagellar RSP3 is an AKAP and reveal that a mutation in the PKA binding domain results in unregulated axonemal PKA activity and inhibition of normal motility.


Granularity impact on hotspot formation and local chemistry in shocked nanostructured RDX.
The nanostructuration of energetic materials results in interesting properties. In particular, the detonation of carbon-rich explosives leads to the formation of nanodiamonds, the size of which is linked to the initial size of the explosive nanograins. This correlation could come from the role played by the granularity in the shock properties, especially the local temperature, which could be enhanced when the shock front crosses the various interfaces and nanoporosities of the material. More generally, the granularity-dependent reactivity also concerns some aspects of the sensitivity of energetic materials and subsequent inhibition or safety issues. In order to investigate this hypothesis, we perform classical and reactive molecular dynamics shock-simulations on cyclotrimethylene trinitramine (RDX), a common high-explosive. We design various nanogranular structures using the body-centered-cubic stacking of nanoparticles shaped as Kelvin cells (truncated octahedra) with slightly-bumped faces. The nanograin size and the bump radius allow to independently control the porosity value and its nanostructure. We show that the global shock properties, especially the temperature, are sensitive to the porosity value but not to the nanograin size. The porosity nanostructure has a local impact, enhancing the temperature heterogeneities between the inter- and intra-grain regions by a few hundred degrees and slowing down the thermal homogenization. For a given porosity, the larger the nanograins, the larger and the hotter the hotspots. In such hotspots, the local chemistry is significantly modified, resulting in a larger reactivity with a quicker formation of some final products. We suggest that the quicker consumption of heteroatoms (namely, H, O, and N) along with higher local temperatures is likely to impact the formation process of solid carbonaceous phases.


Stachyflin and acetylstachyflin, novel anti-influenza A virus substances, produced by Stachybotrys sp. RF-7260. I. Isolation, structure elucidation and biological activities.
Two novel compounds, stachyflin and acetylstachyflin, have been isolated by solid-state fermentation of Stachybotrys sp. RF-7260. The structures of both metabolites, determined by detailed NMR analyses and X-ray crystallographic analysis, are novel with a pentacyclic moiety including cis-fused decalin. The absolute stereochemistry of stachyflins was determined by circular dichroism analysis. Stachyflin showed antiviral activity against influenza A virus (H1N1) in vitro with an IC50 value of 0.003 microM. Acetylstachyflin was about 77-fold less active than stachyflin.


[Interaction of molecular forms of ceruloplasmin with specific membrane receptors from healthy persons and patients with hepatolenticular degeneration].
The ceruloplasmin (CP) interaction with the specific receptor localized on the erythrocyte membrane of healthy donors and of patients with hepatolenticular degeneration (HLD) was studied by using Scatchard analysis. It was found that in patients with the Wilson's disease the CP receptor is unaffected by the pathological process. The molecular mass of the erythrocyte receptor of CP is about 130 kDa. The CP binding to the specific receptor is followed by limited proteolysis of the latter. No CP binding to the receptor occurs in the presence of serine protease inhibitors. The CP-like protein previously detected in the sera of Wilson gene carriers can specifically interact with the CP receptor.


Signs and symptoms of acute mania: a factor analysis.
The major diagnostic classifications consider mania as a uni-dimensional illness. Factor analytic studies of acute mania are fewer compared to schizophrenia and depression. Evidence from factor analysis suggests more categories or subtypes than what is included in the classification systems. Studies have found that these factors can predict differences in treatment response and prognosis. The sample included 131 patients consecutively admitted to an acute psychiatry unit over a period of one year. It included 76 (58%) males. The mean age was 44.05 years (SD = 15.6). Patients met International Classification of Diseases-10 (ICD-10) clinical diagnostic criteria for a manic episode. Patients with a diagnosis of mixed bipolar affective disorder were excluded. Participants were evaluated using the Young Mania Rating Scale (YMRS). Exploratory factor analysis (principal component analysis) was carried out and factors with an eigenvalue > 1 were retained. The significance level for interpretation of factor loadings was 0.40. The unrotated component matrix identified five factors. Oblique rotation was then carried out to identify three factors which were clinically meaningful. Unrotated principal component analysis extracted five factors. These five factors explained 65.36% of the total variance. Oblique rotation extracted 3 factors. Factor 1 corresponding to 'irritable mania' had significant loadings of irritability, increased motor activity/energy and disruptive aggressive behaviour. Factor 2 corresponding to 'elated mania' had significant loadings of elevated mood, language abnormalities/thought disorder, increased sexual interest and poor insight. Factor 3 corresponding to 'psychotic mania' had significant loadings of abnormalities in thought content, appearance, poor sleep and speech abnormalities. Our findings identified three clinically meaningful factors corresponding to 'elated mania', 'irritable mania' and 'psychotic mania'. These findings support the multidimensional nature of manic symptoms. Further evidence is needed to support the existence of corresponding clinical subtypes.


Hydrolysis of transforming growth factor-alpha by cell-surface peptidases in vitro.
Human transforming growth factor-alpha (h-TGF alpha), a 50-amino acid residue peptide, was incubated with some purified cell-surface peptidases and with renal microvillar membranes prepared from pig and rat. Hydrolysis was monitored by h.p.l.c. and activity by a biological assay. Prolonged incubation with relatively large amounts of endopeptidase-24.11, aminopeptidase N and peptidyl dipeptidase A (angiotensin-converting enzyme) caused no observable hydrolysis and no detectable loss of biological activity. Incubation with pig renal microvilli also failed to degrade the peptide. In contrast, rat renal microvilli readily degraded h-TGF alpha, as did endopeptidase-2, which is located in rat renal and intestinal brush borders, but is absent from pig kidneys. This enzyme degraded about 30 nmol of h-TGF alpha/h per mg of protein. The physiological significance of these results is discussed.


Guidelines for the Use of Deuterium Oxide (D2O) in 1H NMR Metabolomics.
In metabolomics, nuclear magnetic resonance (NMR) spectroscopy allows to identify and quantify compounds in biological samples. The sample preparation generally requires only few steps; however, an indispensable factor is the addition of a locking substance into the biofluid sample, such as deuterium oxide (D2O). While creatinine loss in pure D2O is well-described, the effects of different D2O concentrations on the signal profile of biological samples are unknown. In this work, we investigated the effect of D2O levels in the NMR buffer system in urine samples, in dependence on dwell time and temperature exposition. We reveal a decrease of the urinary creatinine peak area up to 35% after 24 h of dwell time at room temperature (RT) using 25% (v/v) D2O, but only 4% loss using 2.5% D2O. 1H, inverse-gated (IG) 13C, DEPT-HSQC NMR, and mass spectrometry (MS) experiments confirmed a proton-deuterium (H/D) exchange at the CH2. This leads to underestimation of creatinine levels and has an extensive effect when creatinine is used for normalization. This work offers a sample stability examination, depending on the D2O concentration, dwell time, and temperature and enables a method to correct for the successive loss. We propose an equation to correct the creatinine loss for samples prepared with various D2O concentrations and storage temperatures for dwell times up to 24 h. The correction function was validated against an external data set with n = 26 samples. To ensure sufficient creatinine stability in future studies, we suggest that a maximum of 10% D2O should be used at 4 °C or 2.5% D2O at RT, respectively.


HIF1α and HIF2α immunoreactivity in epithelial tissue of primary and recurrent pterygium by immunohistochemical analysis.
Hypoxia-inducible factors (HIFs) are considered to play a significant role in the pathogenesis of pterygium. The aim of this study was to investigate the relative expression or immunoreactivity of HIF1α and HIF2α in the epithelium of primary pterygium, recurrences and healthy conjunctiva. Immunohistochemical staining was performed with antibodies against HIF1α and HIF2α, respectively, on 55/84 primary pterygium specimens, 6/28 recurrences and 20/20 control tissues (healthy conjunctiva). Immunohistochemical staining revealed lower epithelial immunoreactivity of HIF1α and HIF2α in both primary pterygium (11% and 38%) and recurrences (18% and 21%) when compared to healthy conjunctival tissue (46% and 66%). Differences between immunoreactivity of HIF1α and of HIF2α in primary pterygium and controls were each highly significant (p < .001). Within the group of primary pterygium, epithelial immunoreactivity of HIF2α (38%) was significantly higher than that of HIF1α (11%). In recurrent pterygium and healthy conjunctiva, immunoreactivity levels of HIF2α were higher than those of HIF1α as well; however, differences between both isoforms were not significant. Our study shows evidence that the higher expressed epithelial HIF2α, rather than HIF1α, and the balance between both HIF isoforms might be relevant factors associated with pathogenesis of primary pterygium. Modulation of HIF2α levels and activity may thus offer a new therapeutic approach to the treatment of advancing pterygium where the initial stage with its HIF1-peak has already passed.


Posttraumatic Haematuria with Pseudorenal Failure: A Diagnostic Lead for Intraperitoneal Bladder Rupture.
Bladder rupture is a very morbid injury following blunt or penetrating lower abdominal trauma. Prompt diagnosis is crucial to initiate optimal treatment. Intraperitoneal bladder rupture is associated with haematuria and biochemical features of renal failure. Cystogram is diagnostic. Immediate open surgical repair is the main stay of treatment. A case of intraperitoneal rupture diagnosed preoperatively by the presence of haematuria and pseudorenal failure is presented to highlight the association of posttraumatic haematuria and pseudorenal failure in such injuries.


[AbstractA woman with knee pain after a fall in childhood].
A 32-year-old woman presented with pain in her left knee ever since she had fallen down the stairs as a child and had landed on this knee. On the X-ray we saw two large patella fragments. We made the diagnosis of dislocated and subluxated fracture of the patella without consolidation. The previously missed fracture was treated conservatively.


[Gastroenterology in the G-DRG-System 2004].
After a year of preliminary voluntarily introduction of casemix funding in hospitals in 2003 nearly every German hospital will be confronted with lump sump payments on the basis of the G-DRG system for their inpatient care starting from January 2004. To analyse weaknesses referring to gastroenterology services within the G-DRG version 1.0 the German Association for Disorders of the Digestive System and Metabolism (DGVS) and the DRG-Research-Group from the University of Muenster conducted a DRG evaluation project. In the analysis patient data from 16 hospitals were included. As a result of the project recommendations for G-DRG adjustments were generated. Those recommendations were implemented in the advancement to G-DRG version 2004. Also the International Classification of Diseases (ICD-10) was modified to ICD-10 German Modification. The classification of procedures OPS-301 was revised. The main adjustments to the G-DRG system and both classifications will be presented in this paper.


Primary antifungal prophylaxis in hematopoietic stem cell transplant recipients: clinical implications of recent studies.
Infectious complications after hematopoietic stem cell transplant, especially those caused by fungi, contribute to poor outcomes. Availability of new antifungal drugs has led to creative attempts to prevent infections caused by both yeasts and molds with new antifungal prophylaxis strategies; these studies will be reviewed. Trials evaluating mold-active azoles, posaconazole and voriconazole, demonstrate that invasive aspergillosis infections can be prevented by prophylactic therapy in patients with hematologic malignancies and after allogeneic hematopoietic stem cell transplant. However, issues arise regarding appropriate dosing and levels, drug interactions, and the importance of breakthrough infections caused by organisms that demonstrate microbiologic resistance. Other recent studies evaluating efficacy and safety of aerosolized lipid formulations of amphotericin B demonstrate promise with directed delivery to the lungs; more long-term follow-up safety information is needed to be assured that this strategy is safe late after allogeneic hematopoietic stem cell transplant, in the setting of acute and chronic graft vs. host disease. Clinicians should consider using new mold-active azole drugs for prophylaxis instead of fluconazole in allogeneic hematopoietic stem cell transplant recipients that have high risks due to prolonged neutropenia and severe graft vs. host disease. More investigations are needed to support other preventive strategies that utilize diagnostic tests and aerosolized delivery of amphotericin B formulations.


Metabolism of leukotriene D4 by porcine alveolar macrophages.
Porcine alveolar macrophages (AM), as we reported in 1981, inactivate slow-reacting substance of anaphylaxis (SRS-A) in a time- and cell-number-dependent fashion. In the present study, metabolism of synthetic leukotriene D4 (LTD4) to leukotriene E4 (LTE4) by AM was demonstrated (mean conversion, 216 pmoles LTD/10(7) AM/60 min). The metabolism was inhibited by cooling or by removal of calcium ions plus addition of EDTA but not by dinitrophenol or puromycin, lipoxygenase inhibitors, the peroxidase inhibitor aminotriazole, or the hydroxyl ion scavenger benzoic acid. Similarly, although the addition of catalase plus superoxide dismutase significantly augmented (169 +/- 25% control) SRS release from dispersed porcine lung cells, the presence of these enzymes did not prevent LTD4 metabolism by AM. Inhibitors of gamma-glutamyl transpeptidase were also without effect but the dipeptidase inhibitors L-cysteine and dithiothreitol significantly reduced the conversion, as did pretreatment of AM with thermolysin (100 micrograms/ml). These data indicate that an AM dipeptidase, which may at least partly be located on the cell surface, converts LTD4 to the less bioactive LTE4.


Epitope-specific antibody and suppression of autoantibody responses against a hybrid self protein.
This study addresses the relationship of epitope-specific Ab responses and alternative autoantibody responses in a model system in which an antigenized self protein serves as the carrier for a defined heterologous B cell epitope. Ubiquitin, a nonimmunogenic self protein, was engineered to present heterologous B and T cell epitopes in the recombinant molecule. Fusion to the C terminus introduced a universal T cell epitope from a Mycobacterium tuberculosis Ag. The B cell epitope was created by inserting a 12-residue loop sequence of HIV-1 gp120 at a surface-exposed position of ubiquitin. These modifications preserved the ubiquitin fold, allowing a new conformational epitope to be presented among native self epitopes. Mice immunized with the hybrid protein bearing only the mycobacterial T cell epitope elicited a strong autoantibody response to native ubiquitin. In contrast, antisera elicited against hybrid ubiquitin presenting the HIV B cell epitope reacted specifically with the foreign epitope but not with native ubiquitin. Absence of autoantibody in the response was attributed to poor competition of autoreactive B cells for limiting T cell help. Both types of responses were associated with Th responses to defined epitopes of the ubiquitin hybrid protein. These results may have implications for a tolerance mechanism dependent on B-T cell cooperation.


Has the recessive hypothesis for susceptibility to insulin-dependent diabetes mellitus been firmly and unequivocally rejected?
An analysis of an implied assumption, that of Hardy-Weinberg equilibrium, reveals that the single locus, recessive allele hypothesis for susceptibility to insulin-dependent diabetes mellitus has not been firmly and unequivocally rejected by the method of Rotter et al.


Relationship between mean daily energy intake and frequency of consumption of out-of-home meals in the UK National Diet and Nutrition Survey.
Out-of-home meals have been characterised as delivering excessively large portions that can lead to high energy intake. Regular consumption is linked to weight gain and diet related diseases. Consumption of out-of-home meals is associated with socio-demographic and anthropometric factors, but the relationship between habitual consumption of such meals and mean daily energy intake has not been studied in both adults and children in the UK. We analysed adult and child data from waves 1-4 of the UK National Diet and Nutrition Survey using generalized linear modelling. We investigated whether individuals who report a higher habitual consumption of meals out in a restaurant or café, or takeaway meals at home had a higher mean daily energy intake, as estimated by a four-day food diary, whilst adjusting for key socio-demographic and anthropometric variables. Adults who ate meals out at least weekly had a higher mean daily energy intake consuming 75-104 kcal more per day than those who ate these meals rarely. The equivalent figures for takeaway meals at home were 63-87 kcal. There was no association between energy intake and frequency of consumption of meals out in children. Children who ate takeaway meals at home at least weekly consumed 55-168 kcal more per day than those who ate these meals rarely. Additionally, in children, there was an interaction with socio-economic position, where greater frequency of consumption of takeaway meals was associated with higher mean daily energy intake in those from less affluent households than those from more affluent households. Higher habitual consumption of out-of-home meals is associated with greater mean daily energy intake in the UK. More frequent takeaway meal consumption in adults and children is associated with greater daily energy intake and this effect is greater in children from less affluent households. Interventions seeking to reduce energy content through reformulation or reduction of portion sizes in restaurants, cafés and takeaways could potentially lead to reductions in mean daily energy intake, and may reduce inequalities in health in children.


Phenotypic characterization of murine tumor-infiltrating T lymphocytes.
Tumor infiltrating lymphocytes (TIL) can be isolated from solid tumors and selectively expanded in long term culture with IL-2 and autologous irradiated tumor. Such long term cultured cells express anti-tumor activity in vitro, mediate the regression of established tumor in murine models of cancer, and have been used for the treatment of cancer in humans. We have characterized freshly isolated mouse Thy-1+ TIL populations, as well as long term TIL cultures, from several different C57BL/6 (B6) tumors. Freshly isolated Thy-1+ TIL include both CD4+ and CD8+ cells, as well as cells bearing NK markers. These cells are predominantly TCR alpha beta+, with a smaller population of TCR gamma delta+ cells. The TCR alpha beta+ cells expressed a broad distribution of V beta phenotypes that was statistically different from that expressed in normal B6 splenic Thy-1+ cells or CD8+ cells, presumably reflecting in vivo selection in the host anti-tumor response. NK cells are present in these tumors at a greater frequency than noted in splenic T cells. Cultured TIL populations rapidly became exclusively Thy-1+/CD8+/CD4- and TCR alpha beta+/gamma delta-. Individual long term TIL populations initially expressed multiple V beta products, but rapidly restricted their V beta expression, frequently expressing a single dominant V beta. The identity of this dominant V beta varied among different TIL lines, but the overall representation of V beta phenotypes in these cultures was statistically different from that seen in Thy-1+ or CD8+ splenocytes. No statistical difference was noted between lines derived from antigenically distinct tumors. The selection of tumor specific T cells in vitro is therefore not reflected in any simple predominance of V beta usage. The complexity of TCR usage in the anti-tumor response may result from the involvement of multiple alpha- and beta-chain regions in the response to a single antigenic determinant, or may reflect multiple antigenic determinants expressed on a single syngeneic tumor.


An experimental protocol of the model to quantify traction applied to the retina by vitreous cutters.
The purpose of this study was to use a novel method to quantify the traction applied to the retina during vitrectomy. Five 20-gauge electric cutters were used. Fresh porcine eyes were positioned in a specially developed holder and transfixed to the retinal layers with a microwire, and the other end was fixed to the load cell of a strain gauge. The cutter to be assessed was introduced into the eye by a micromanipulator at a 45 degrees angle adjacent to the retina. The traction force was evaluated when the cutter was at 3 and 5 mm from the retina. As control, the experiment was repeated in eyes filled with water, and the results were compared to those when the eyes had vitreous gel. Results from the eyes with vitreous gel indicate that retinal traction increased with increasing aspiration vacuum (7.90 dyn for each 100 mm Hg increased; P < 0.05) and proximity to the retina (2.17 dyn; P < 0.05) and decreased with increasing cut rate (2.51 dyn for each 500 cuts per minute increased; P < 0.05). In all eyes filled with water, traction was not observed. The present study establishes a new reproducible technique to quantify vitreoretinal traction during vitrectomy and demonstrates that the effects of aspiration, distance from the retina, and cut rate are crucial factors in the amount of retinal traction created by vitreous cutters.


Hierarchical role for transcription factors and chromatin structure in genome organization along adipogenesis.
The three dimensional folding of mammalian genomes is cell type specific and difficult to alter suggesting that it is an important component of gene regulation. However, given the multitude of chromatin-associating factors, the mechanisms driving the colocalization of active chromosomal domains and the role of this organization in regulating the transcription program in adipocytes are not clear. Analysis of genome-wide chromosomal associations revealed cell type-specific spatial clustering of adipogenic genes in 3T3-L1 cells. Time course analysis demonstrated that the adipogenic 'hub', sampled by PPARγ and Lpin1, undergoes orchestrated reorganization during adipogenesis. Coupling the dynamics of genome architecture with multiple chromatin datasets indicated that among all the transcription factors (TFs) tested, RXR is central to genome reorganization at the beginning of adipogenesis. Interestingly, at the end of differentiation, the adipogenic hub was shifted to an H3K27me3-repressive environment in conjunction with attenuation of gene transcription. We propose a stage-specific hierarchy for the activity of TFs contributing to the establishment of an adipogenic genome architecture that brings together the adipogenic genetic program. In addition, the repositioning of this network in a H3K27me3-rich environment at the end of differentiation may contribute to the stabilization of gene transcription levels and reduce the developmental plasticity of these specialized cells. All sequence data reported in this paper have been deposited at GEO (http://www.ncbi.nlm.nih.gov/geo/) (GSE92475).


Recovery Following Acute Exacerbations of Chronic Obstructive Pulmonary Disease - A Review.
Acute exacerbations are associated with disease progression, hospital admission and death in people with chronic obstructive pulmonary disease (COPD). The detrimental outcomes associated with acute exacerbations highlights a need to understand the time course of recovery following acute exacerbation of COPD (AECOPD) so that effective and timely interventions can be provided. The aim of this narrative review was to describe the natural recovery in physiology, symptoms and function following AECOPD. Substantial recovery of lung function and airway inflammation occurs in the first week after onset of an AECOPD, whilst systemic inflammatory markers may take up to two weeks to recover. Symptoms generally improve over the first 14 days, however marked variation is evident between studies and individuals. There are limited data regarding the time course of recovery for functional capacity, quality of life and strength. In a small number of patients (<10%) recovery of lung function and symptoms has not occurred by three months. Features of patients at risk of a prolonged recovery following AECOPD include older age, more severe lung disease, presence of chronic bronchitis, lower body mass index and more chronic dyspnoea. Exacerbation features associated with prolonged recovery are symptoms of the common cold at exacerbation onset, evidence of viral infection, more severe dyspnoea during the exacerbation and persistent systemic inflammation. In clinical practice efforts should be made to recognise prolonged recovery, which puts patients at risk of poor outcomes, and to address the consequences of AECOPD including physical inactivity and skeletal muscle weakness. Whether delivery of specific interventions at distinct time points in the recovery process can enhance recovery remains to be determined.


Fetal hypothalamic-pituitary adrenal (HPA) development and activation as a determinant of the timing of birth, and of postnatal disease.
Birth in most animal species is triggered by the fetus through activation of the fetal hypothalamic-pituitary-adrenal (HPA) axis. Preterm birth, may be associated with precocious activation of fetal HPA function, reflecting the fetal response to an adverse intrauterine environment. There is a progressive and concurrent increase of ACTH1-39 and cortisol (F) in the circulation of fetal sheep during the last 15-20 days of pregnancy (term, day 145-150) associated with increased expression of hypothalamic CRH pituitary POMC and adrenal ACTH receptor and steroidogenic enzymes, particularly P450 C17. Similar changes occur with fetal hypoxemia. Negative feedback is ameliorated by decreased pituitary and hypothalamic glucocorticoid receptor, increased CBG, and altered fetal pituitary 11B-hydroxysteroid dehydrogenase type 1. Repeated fetal hypoxemia, diminishes the fetal-pituitary ACTH response, but increases fetal adrenal responsiveness. Fetuses exposed to maternal glucocorticoid in late gestation are growth restricted with altered postnatal HPA responsiveness and glycemic responses that reproduce the insulin resistance of type 2 diabetes. We conclude that the level of fetal HPA activity is crucial not only for determining gestation length, but also predicts pathophysiologic adjustment in later life.


Improved flow cytometric analysis of leukocyte subsets: simultaneous identification of five cell subsets using two-color immunofluorescence.
Flow cytometric analysis of human peripheral blood leukocytes has typically been achieved by staining multiple aliquots of the same sample with fluorescent reagents specific for cell subsets of interest. Spectrally discrete fluorochrome tags have been developed for applications in which identification of multiple subsets (e.g., T and B cells) or of subsets not uniquely identified by a single reagent (e.g., activated T cells) requires use of multiple reagents per aliquot. Extension of this approach to more than two reagents per aliquot has led to multicolor methods requiring dual laser excitation and complex instrumentation. We describe an alternative two-color method using commercially available reagents that allows simultaneous identification of five discrete immune cell subsets using only a single excitation source. The technique uses dilution of commercial fluorochrome-labeled reagents with competing unlabeled reagents to selectively produce discrete fluorescence intensity profiles for cell subsets that would otherwise display overlapping or indistinguishable profiles when stained with reagents bearing the same fluorochrome. For example, the fluorescence intensity of phycoerythrin-labeled helper T (Th) cells can be adjusted to be distinct from that of phycoerythrin-labeled suppressor T (Ts) cells. Extending this technique to two colors, we have used a combination of seven different monoclonal antibodies to simultaneously quantify Th, Ts, B cells, natural killer cells, and monocytes in a single aliquot. An additional advantage of this approach is the ability to more accurately quantify "null" cells. Adjustment of fluorescence intensity profiles of different cell subsets by this method is applicable to flow cytometric analysis of a wide variety of cell types. The technique significantly extends the analytical capacity of flow cytometry without significantly increasing the complexity of the instrumentation required.


Caries experience and treatment response among groups of Finnish, German and Russian children. A retrospective study between 1995 and 1989.
This study was designed to describe caries experience and treatment response in groups of children in three countries--Finland, Germany and Russia--with different systems of public dental care. Information concerning permanent teeth (carious and filled surfaces, type of restorative treatment and applied sealants) was collected retrospectively from existing dental records of 12-year-old children who had been treated in dental practices over six years, retrospectively between 1995 and 1989. Prior to conducting the study, research meetings were held among all participating personnel to standardise data collection and terminology as much as possible. The analysis of variance (ANOVA) was used to test the difference between the means of the various types of data derived from the three countries. Sealants were most commonly used in Finland at a rate of 5.0/child and 3.3 in Germany, but only for a few children in Russia (0.1). Caries increments (cumulative figure of primary and secondary caries surfaces over the six follow-up years of the study) differed very much between these countries. It was very high in Russia (7.4) and in Germany (5.3), but low in Finland (1.6). All the caries surfaces were filled in Finland and in Germany; in Russia, the cumulated number of filled surfaces was smaller than the respective number of caries surfaces (6.4/7.4). The most commonly used restorative filling material was amalgam in Germany and in Russia, but composite or glass ionomer cements in Finland. The cost effectiveness of sealant application programs should be considered as exemplified by the Finnish dental care system. The data showed a need for preventive programs in Germany, and this has actually been emphasised since 1993. In Russia, there is an urgent need to implement preventive programs for 6-year-old children and also to provide the necessary restorative treatment.


The challenges of antimicrobial resistance in Brazil.
Brazil is a country with continental proportions with high geographic and economic diversity. Despite its medical centers of excellence, antimicrobial resistance poses a major therapeutic challenge. Rates of methicillin-resistant Staphylococcus aureus are up to 60% and are related to an endemic Brazilian clone. Local resistance to vancomycin in Enterococci was first related to Enterococcus faecalis, which differs from European and American epidemiology. Also, local Klebsiella pneumoniae and Escherichia coli isolates producing extended-spectrum β-lactamases have a much higher prevalence (40%-50% and 10%-18%, respectively). Carbapenem resistance among the enterobacteriaceae group is becoming a major problem, and K. pneumoniae carbapenemase isolates have been reported in different states. Among nonfermenters, carbapenem resistance is strongly related to SPM-1 (Pseudomonasaeruginosa) and OXA-23 (Acinetobacter baumannii complex) enzymes, and a colistin-only susceptible phenotype has also emerged in these isolates, which is worrisome. Local actions without loosing the global resistance perspective will demand multidisciplinary actions, new policies, and political engagement.


On the origin of self-incompatibility haplotypes: transition through self-compatible intermediates.
Self-incompatibility (SI) in flowering plants entails the inhibition of fertilization by pollen that express specificities in common with the pistil. In species of the Solanaceae, Rosaceae, and Scrophulariaceae, the inhibiting factor is an extracellular ribonuclease (S-RNase) secreted by stylar tissue. A distinct but as yet unknown gene (provisionally called pollen-S) appears to determine the specific S-RNase from which a pollen tube accepts inhibition. The S-RNase gene and pollen-S segregate with the classically defined S-locus. The origin of a new specificity appears to require, at minimum, mutations in both genes. We explore the conditions under which new specificities may arise from an intermediate state of loss of self-recognition. Our evolutionary analysis of mutations that affect either pistil or pollen specificity indicates that natural selection favors mutations in pollen-S that reduce the set of pistils from which the pollen accepts inhibition and disfavors mutations in the S-RNase gene that cause the nonreciprocal acceptance of pollen specificities. We describe the range of parameters (rate of receipt of self-pollen and relative viability of inbred offspring) that permits the generation of a succession of new specificities. This evolutionary pathway begins with the partial breakdown of SI upon the appearance of a mutation in pollen-S that frees pollen from inhibition by any S-RNase presently in the population and ends with the restoration of SI by a mutation in the S-RNase gene that enables pistils to reject the new pollen type.