[HERS study disturbs hormonal replacement therapy].
Cardiovascular protection of hormonal replacement therapy was considered a fact. The effects of estrogens on lipid levels and vascular health gave biological support to estrogen cardioprotection. The recently published HERS study showing no protective effects of estrogen and progesterone replacement therapy on the risk of myocardial infarction or coronary deaths is provoking perplexity. These surprising results may have several causes such as the use of progesterone, the associated use of cardioprotective agents or the short observation period. However, the study results scope is restricted to secondary prevention. These cannot be extrapolated to frequent conditions of postmenopausal women such as primary prevention or sequential schemes. Likewise, this investigation underscores the importance of a critical and continued evaluation of apparently well established therapeutic approaches.


Photo-Programmed Deformations in Rigid Liquid Crystalline Polymers Triggered by Body Temperature.
Deformations triggered by body heat are desirable in the context of shape-morphing applications because, under the majority of circumstances, the human body maintains a higher temperature than that of its surroundings. However, at present, this bioenergy-triggered action is primarily limited to soft polymeric networks. Thus, herein, the programming of body temperature-triggered deformations into rigid azobenzene-containing liquid crystalline polymers (azo-LCPs) with a glass-transition temperature of 100 °C is demonstrated. To achieve this, a mechano-assisted photo-programming strategy is used to create a metastable state with room-temperature stable residual stress, which is induced by the isomerization of azobenzene. The programmed rigid azo-LCP can undergo large-amplitude body temperature-triggered shape changes within minutes and can be regenerated without any performance degradation. By changing the programming photomasks and irradiation conditions employed, various 2D to 3D shape-morphing architectures, including folded clips, inch-worm structures, spiral structures, and snap-through motions are achieved. When programmed with polarized light, the proposed strategy results in domain-selective activation, generating designed characteristics in multi-domain azo-LCPs. The reported strategy is therefore expected to broaden the applications of azo-LCPs in the fields of biomedical and flexible microelectronic devices.


3D Printed Conformal Strain and Humidity Sensors for Human Motion Prediction and Health Monitoring via Machine Learning.
Wearable sensors have garnered considerable attention due to their flexibility and lightweight characteristics in the realm of healthcare applications. However, developing robust wearable sensors with facile fabrication and good conformity remains a challenge. In this study, a conductive graphene nanoplate-carbon nanotube (GC) ink is synthesized for multi jet fusion (MJF) printing. The layer-by-layer fabrication process of MJF not only improves the mechanical and flame-retardant properties of the printed GC sensor but also bolsters its robustness and sensitivity. The direction of sensor bending significantly impacts the relative resistance changes, allowing for precise investigations of joint motions in the human body, such as those of the fingers, wrists, elbows, necks, and knees. Furthermore, the data of resistance changes collected by the GC sensor are utilized to train a support vector machine with a 95.83% accuracy rate for predicting human motions. Due to its stable humidity sensitivity, the sensor also demonstrates excellent performance in monitoring human breath and predicting breath modes (normal, fast, and deep breath), thereby expanding its potential applications in healthcare. This work opens up new avenues for using MJF-printed wearable sensors for a variety of healthcare applications.


Cardiovascular disease risk factors and n-3 fatty acid status in the adult population of James Bay Cree.
Canadian native populations, which traditionally consume large amounts of fish, have lower rates of mortality from heart disease than do Canadian nonnative populations, which have low fish intakes. Fish oils rich in n-3 fatty acids may have a protective effect against cardiovascular disease (CVD) risk factors. The purposes were to examine the profile of plasma phospholipid concentrations of the n-3 fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) among James Bay Cree and to verify the relation between these concentrations and CVD risk factors. The study population consisted of 917 subjects aged 18-74 y who participated in the 1991 Santé Québec Health Survey. Data were obtained through home interviews and clinic visits. Plasma samples were analyzed for phospholipid fatty acid composition. The mean fish consumption on the day before the survey was 60 g among the adult Cree population. Expressed as a percentage of total fatty acids, relative concentrations of EPA and DHA were 0.65% and 2.80%, respectively. n-3 Fatty acids were higher among coastal residents than among inland residents. A positive association was observed between plasma HDL and n-3 fatty acids. EPA and EPA+DHA were inversely associated with triacylglycerols. Among subjects aged 50-74 y, an inverse association between EPA and EPA:AA and total:HDL cholesterol was observed. n-3 Fatty acids may favorably influence some CVD risk factors. The Cree population must be encouraged to maintain their traditional fish-based diet, which may be one of the factors protecting them against mortality from CVD.


Isolation and molecular characterisation of chicken parvovirus from Brazilian flocks with enteric disorders.
1. The presence of parvovirus in chickens with enteric disease was investigated in commercial flocks in Brazil. 2. The intestinal contents of chickens exhibiting clinical signs of diarrhoea, weight loss or mortality were examined, and chicken parvovirus (chPV) was identified using a polymerase chain reaction (PCR) assay. The samples were sequenced and inoculated into specific-pathogen-free (SPF) embryonated eggs to isolate the virus. 3. Necropsies showed that the embryos were dwarfish, haemorrhagic and oedematous. The presence of chPV was confirmed by PCR and DNA sequencing. 4. The molecular characterisation of chPV strains circulating in the Brazilian flocks showed that they were genetically related to sequences from North America, Europe and Asia. Phylogenetic analyses clustered the Brazilian chPV sequences with those from Europe (Croatia, Hungary) and Asia (South Korea). 5. This study is the first report of the molecular characterisation of chPV circulating in the commercial flocks in Brazil and indicates high genetic similarity with chPV sequences from around the world.


A systematic review and meta-analysis on the prevalence of Dupuytren disease in the general population of Western countries.
Dupuytren disease is a fibroproliferative disease of palmar fascia of the hand. Its prevalence has been the subject of several reviews; however, an accurate description of the prevalence range in the general population--and of the relation between age and disease--is lacking. Embase and PubMed were searched using database-specific Medical Subject Headings; titles and abstracts were searched for the words "Dupuytren," "incidence," and "prevalence." Two reviewers independently assessed the articles using inclusion and exclusion criteria, and rated the included studies with a quality assessment instrument. In a meta-analysis, the median prevalence, as a function of age by sex, was estimated, accompanied by 95 percent prediction intervals. The observed heterogeneity in prevalence was investigated with respect to study quality and geographic location. Twenty-three of 199 unique identified articles were included. The number of participants ranged from 37 to 97,537, and age ranged from 18 to 100 years. Prevalence varied from 0.6 to 31.6 percent. The quality of studies differed but could not explain the heterogeneity among studies. Mean prevalence was estimated as 12, 21, and 29 percent at ages 55, 65, and 75 years, respectively, based on the relation between age and prevalence determined from 10 studies. The authors describe a prevalence range of Dupuytren disease in the general population of Western countries. The relation between age and prevalence of Dupuytren disease is given according to sex, including 95 percent prediction intervals. It is possible to determine disease prevalence at a certain age for the total population, and for men and women separately.


Effective Inhibition of TDP-43 Aggregation by Native State Stabilization.
Preventing the misfolding or aggregation of transactive response DNA binding protein with 43 kDa (TDP-43) is the most actively pursued disease-modifying strategy to treat amyotrophic lateral sclerosis and other neurodegenerative diseases. In this work, we provide proof of concept that native state stabilization of TDP-43 is a viable and effective strategy for treating TDP-43 proteinopathies. Firstly, we leveraged the Cryo-EM structures of TDP-43 fibrils to design C-terminal substitutions that disrupt TDP-43 aggregation. Secondly, we showed that these substitutions (S333D/S342D) stabilize monomeric TDP-43 without altering its physiological properties. Thirdly, we demonstrated that binding native oligonucleotide ligands stabilized monomeric TDP-43 and prevented its fibrillization and phase separation in the absence of direct binding to the aggregation-prone C-terminal domain. Fourthly, we showed that the monomeric TDP-43 variant could be induced to aggregate in a controlled manner, which enabled the design and implementation of a high-throughput screening assay to identify native state stabilizers of TDP-43. Altogether, our findings demonstrate that different structural domains in TDP-43 could be exploited and targeted to develop drugs that stabilize the native state of TDP-43 and provide a platform to discover novel drugs to treat TDP-43 proteinopathies.


[Short-term prognosis in cryptogenetic hemoptysis].
149 patients suffering from hemoptysis of unknown origin who presented a normal chest X-ray were followed up, in order to evaluate their prognosis, during a mean period of 12.6 months after the initial study which included fiberoptic bronchoscopy. During this time, 24 patients (16.1%) presented new episodes of hemoptysis without radiological modifications; a second fiberoptic bronchoscopy carried out in 13 of these patients did not contribute with any new diagnostic data. One patient was diagnosed of pulmonary thromboembolism after persistent hemoptysis and two others of bronchial carcinoma 4 and 30 months after the initial fiberoptic bronchoscopy when the chest X-ray had already suffered significant changes. These data confirm that short term prognosis of cryptogenetic hemoptysis is generally favourable when the fiberoptic bronchoscopy is negative. The fact that a high percentage of these patients (37%, 55/149) had previously suffered pulmonary tuberculosis and/or inflammatory-scar type localized endobronchiolar lesions suggests that sequelae of these diseases, not observed in the X-ray, could play an etiological role in the bleeding.


Directional budding of human immunodeficiency virus from monocytes.
Time-lapse cinematography revealed that activated human immunodeficiency virus (HIV)-infected monocytes crawl along surfaces, putting forward a leading pseudopod. Scanning electron micrographs showed monocyte pseudopods associated with spherical structures the size of HIV virions, and transmission electron micrographs revealed HIV virions budding from pseudopods. Filamentous actin (F-actin) was localized by electron microscopy in the pseudopod by heavy meromyosin decoration. Colocalization of F-actin and p24 viral antigen by light microscopy immunofluorescence indicated that F-actin and virus were present on the same pseudopod. These observations indicate that monocytes produce virus from a leading pseudopod. We suggest that HIV secretion at the leading edges of donor monocytes/macrophages may be an efficient way for HIV to infect target cells.


Epidermal growth factor increases coactivation of the androgen receptor in recurrent prostate cancer.
Growth of normal and neoplastic prostate is mediated by the androgen receptor (AR), a ligand-dependent transcription factor activated by high affinity androgen binding. The AR is highly expressed in recurrent prostate cancer cells that proliferate despite reduced circulating androgen. In this report, we show that epidermal growth factor (EGF) increases androgen-dependent AR transactivation in the recurrent prostate cancer cell line CWR-R1 through a mechanism that involves a post-transcriptional increase in the p160 coactivator transcriptional intermediary factor 2/glucocorticoid receptor interacting protein 1 (TIF2/GRIP1). Site-specific mutagenesis and selective MAPK inhibitors linked the EGF-induced increase in AR transactivation to phosphorylation of TIF2/GRIP1. EGF signaling increased the coimmunoprecipitation of TIF2 and AR. AR transactivation and its stimulation by EGF were reduced by small interfering RNA inhibition of TIF2/GRIP1 expression. The data indicate that EGF signaling through MAPK increases TIF2/GRIP1 coactivation of AR transactivation in recurrent prostate cancer.


Why did ancient people have atherosclerosis?: from autopsies to computed tomography to potential causes.
Computed tomographic findings of atherosclerosis in the ancient cultures of Egypt, Peru, the American Southwest and the Aleutian Islands challenge our understanding of the fundamental causes of atherosclerosis. Could these findings be true? Is so, what traditional risk factors might be present in these cultures that could explain this apparent paradox? The recent computed tomographic findings are consistent with multiple autopsy studies dating as far back as 1852 that demonstrate calcific atherosclerosis in ancient Egyptians and Peruvians. A nontraditional cause of atherosclerosis that could explain this burden of atherosclerosis is the microbial and parasitic inflammatory burden likely to be present in ancient cultures inherently lacking modern hygiene and antimicrobials. Patients with chronic systemic inflammatory diseases of today, including systemic lupus erythematosus, rheumatoid arthritis, and human immunodeficiency virus infection, experience premature atherosclerosis and coronary events. Might the chronic inflammatory load of ancient times secondary to infection have resulted in atherosclerosis? Smoke inhalation from the use of open fires for daily cooking and illumination represents another potential cause. Undiscovered risk factors could also have been present, potential causes that technologically cannot currently be measured in our serum or other tissue. A synthesis of these findings suggests that a gene-environmental interplay is causal for atherosclerosis. That is, humans have an inherent genetic susceptibility to atherosclerosis, whereas the speed and severity of its development are secondary to known and potentially unknown environmental factors.


Fatal haemorrhagic infarct in an infant with homocystinuria.
Thrombotic and thromboembolic complications are the main causes of morbidity and mortality in patients with homocystinuria. However, it is unusual for thrombosis and infarction to be the presenting feature leading to investigation for homocystinuria and cerebrovascular lesions in the first year of life. We describe a previously healthy 6-month-old infant who presented with a large middle-cerebral-artery territory infarction and died of massive brain swelling. Homocystinuria due to cystathionine beta-synthase (CBS) deficiency was diagnosed by metabolite analysis and confirmed by enzymatic activity measurement in a postmortem liver biopsy. Homocystinuria should be considered in the differential diagnosis of venous or arterial thrombosis, regardless of age, even in the absence of other common features of the disease. We recommend systematic metabolic screening for hyperhomocysteinemia in any child presenting with vascular lesions or premature thromboembolism.


Thymine metabolism in Pseudomonas aeruginosa strain 1: the presence of a salvage pathway.
Exogenous thymine was found to be taken up very slowly by Pseudomonas aeruginosa in comparison to other pyrimidines, and most of it was catabolized by the cell. The existence of a functional, although inefficient, thymine salvage pathway was demonstrated and this pathway operated more effectively when de novo thymidine nucleotide biosynthesis was inhibited by trimethoprim or methotrexate. The mechanism of thymine salvage by P. aeruginosa appears to be different from that of Escherichia coli and Pseudomonas acidovorans as thymidine was not incorporated into the DNA. Like P. acidovorans, P. aeruginosa lacked thymidine phosphorylase activity. Unsuccessful attempts were made to isolate thymine auxotrophs.


Ad libitum feeding triggers puberty onset associated with increases in arcuate Kiss1 and Pdyn expression in growth-retarded rats.
Increasing evidence shows that puberty onset is largely dependent on body weight rather than chronological age. To investigate the mechanism involved in the energetic control of puberty onset, the present study examined effects of chronic food restriction during the prepubertal period and the resumption of ad libitum feeding for 24 and 48 h on estrous cyclicity, Kiss1 (kisspeptin gene), Tac3 (neurokinin B gene) and Pdyn (dynorphin A gene) expression in the hypothalamus, luteinizing hormone (LH) secretion and follicular development in female rats. When animals weighed 75 g, they were subjected to a restricted feeding to retard growth to 70-80 g by 49 days of age. Then, animals were subjected to ad libitum feeding or remained food-restricted. The growth-retarded rats did not show puberty onset associated with suppression of both Kiss1 and Pdyn expression in the arcuate nucleus (ARC). 24-h ad libitum feeding increased tonic LH secretion and the number of Graafian and non-Graafian tertiary follicles with an increase in the numbers of ARC Kiss1- and Pdyn-expressing cells. 48-h ad libitum feeding induced the vaginal proestrus and a surge-like LH increase with an increase in Kiss1-expressing cells in the anteroventral periventricular nucleus (AVPV). These results suggest that the negative energy balance causes pubertal failure with suppression of ARC Kiss1 and Pdyn expression and then subsequent gonadotropin secretion and ovarian function, while the positive energetic cues trigger puberty onset via an increase in ARC Kiss1 and Pdyn expression and thus gonadotropin secretion and follicular development in female rats.


Reduced Proteolipid Protein 2 promotes endoplasmic reticulum stress-related apoptosis and increases drug sensitivity in acute myeloid leukemia.
The Proteolipid Protein 2 (PLP2), a protein in the Endoplasmic Reticulum (ER) membrane, has been reported to be highly expressed in various tumors. Previous studies have demonstrated that the reduced PLP2 can induce apoptosis and autophagy through ER stress-related pathways, leading to a decreased proliferation and aggressiveness. However, there is no research literature on the role of PLP2 in Acute Myeloid Leukemia (AML). PLP2 expression, clinical data, genetic mutations, and karyotype changes from GEO, TCGA, and timer2.0 databases were analyzed through the R packages. The possible functions and pathways of cells were explored through GO, KEGG, and GSEA enrichment analysis using the clusterProfiler R package. Immuno-infiltration analysis was conducted using the Cibersort algorithm and the Xcell R package. RT-PCR and western blot techniques were employed to identify the PLP2 expression, examine the knockdown effects in THP-1 cells, and assess the expression of genes associated with endoplasmic reticulum stress and apoptosis. Flow cytometry was utilized to determine the apoptosis and survival rates of different groups. PLP2 expression was observed in different subsets of AML and other cancers. Enrichment analyses revealed that PLP2 was involved in various tumor-related biological processes, primarily apoptosis and lysosomal functions. Additionally, PLP2 expression showed a strong association with immune cell infiltration, particularly monocytes. In vitro, the knockdown of PLP2 enhanced endoplasmic reticulum stress-related apoptosis and increased drug sensitivity in THP-1 cells. PLP2 could be a novel therapeutic target in AML, in addition, PLP2 is a potential endoplasmic reticulum stress regulatory gene in AML.


The immunoglobulin octanucleotide: independent activity and selective interaction with enhancers.
The thymidine kinase (tk) promoter of herpes simplex virus includes an octanucleotide sequence motif (ATTTGCAT) that is also an essential component of immunoglobulin kappa gene promoters. In the absence of an enhancer, tk promoter derivatives that contain this element support a higher rate of transcription than those that lack it. The action of the kappa enhancer augments that of the octanucleotide in B lymphoid cells; when both elements are present, tk promoter activity is increased by more than an order of magnitude. In contrast, the presence of the octanucleotide in this promoter markedly reduces its response to a nonimmunoglobulin enhancer. These results suggest that the octanucleotide may mediate a selective interaction among promoters and enhancers.


Hierarchical bayesian modeling of pharmacophores in bioinformatics.
One of the key ingredients in drug discovery is the derivation of conceptual templates called pharmacophores. A pharmacophore model characterizes the physicochemical properties common to all active molecules, called ligands, bound to a particular protein receptor, together with their relative spatial arrangement. Motivated by this important application, we develop a Bayesian hierarchical model for the derivation of pharmacophore templates from multiple configurations of point sets, partially labeled by the atom type of each point. The model is implemented through a multistage template hunting algorithm that produces a series of templates that capture the geometrical relationship of atoms matched across multiple configurations. Chemical information is incorporated by distinguishing between atoms of different elements, whereby different elements are less likely to be matched than atoms of the same element. We illustrate our method through examples of deriving templates from sets of ligands that all bind structurally related protein active sites and show that the model is able to retrieve the key pharmacophore features in two test cases.


Anatomic and pathologic spectrum of pituitary infundibulum lesions.
The pathologic spectrum of pituitary infundibulum disease is diverse. We reviewed 65 infundibular lesions in 44 adult and 21 pediatric patients and summarized their imaging features and clinical presentation. The spectrum of pathology involving the pituitary infundibulum is broad yet distinct from other pathology in the sella and parasellar region. Pituitary stalk lesions can be grouped into three categories: congenital and developmental, inflammatory and infectious, and neoplastic. Knowledge of the imaging appearance of diseases specific to adults and to children is important for accurate diagnosis and treatment.


[Technical recommendations for the electroencephalogram (EEG) recording in suspected brain death].
Brazilian Clinical Neurophysiology Society guidelines and pertaining comments concerning electroencephalogram (EEG) recording in suspected brain death are presented. EEG is not intended as a substitute, rather as a complement to neurologic evaluation.


CellCharter reveals spatial cell niches associated with tissue remodeling and cell plasticity.
Tissues are organized in cellular niches, the composition and interactions of which can be investigated using spatial omics technologies. However, systematic analyses of tissue composition are challenged by the scale and diversity of the data. Here we present CellCharter, an algorithmic framework to identify, characterize, and compare cellular niches in spatially resolved datasets. CellCharter outperformed existing approaches and effectively identified cellular niches across datasets generated using different technologies, and comprising hundreds of samples and millions of cells. In multiple human lung cancer cohorts, CellCharter uncovered a cellular niche composed of tumor-associated neutrophil and cancer cells expressing markers of hypoxia and cell migration. This cancer cell state was spatially segregated from more proliferative tumor cell clusters and was associated with tumor-associated neutrophil infiltration and poor prognosis in independent patient cohorts. Overall, CellCharter enables systematic analyses across data types and technologies to decode the link between spatial tissue architectures and cell plasticity.


A neural network approach for bone fracture healing assessment.
An approach based on auscultatory percussion, a technique used by some orthopedists both for bone fracture detection and bone fracture healing assessment, is described. Low-frequency, low-intensity mechanical power, very much like the finger tap of orthopedists, is used to evaluate the vibrational response of the bone. The novel element is the data processing, which incorporates specialized preprocessing and a neural network for estimating fractured bone strength. In addition, a new mathematical model for the vibrational response of a fractured limb, which provides data to design and test the neural network processing scheme, is presented. An experimental procedure is described for acquiring real data from animal and human fractures in a form necessary for neural network input.


Use of 16S rRNA gene-targeted group-specific primers for real-time PCR analysis of predominant bacteria in human feces.
16S rRNA gene-targeted group-specific primers were designed and validated for specific detection and quantification of the Clostridium leptum subgroup and the Atopobium cluster. To monitor the predominant bacteria in human feces by real-time PCR, we used these specific primers together with four sets of group-specific primers for the Clostridium coccoides group, the Bacteroides fragilis group, Bifidobacterium, and Prevotella developed in a previous study (T. Matsuki, K. Watanabe, J. Fujimoto, Y. Miyamoto, T. Takada, K. Matsumoto, H. Oyaizu, and R. Tanaka, Appl. Environ. Microbiol. 68:5445-5451, 2002). Examination of DNA extracted from the feces of 46 healthy adults showed that the C. coccoides group was present in the greatest numbers (log10 10.3 +/- 0.3 cells per g [wet weight] [average +/- standard deviation]), followed by the C. leptum subgroup (log10 9.9 +/- 0.7 cells per g [wet weight]), the B. fragilis group (log10 9.9 +/- 0.3 cells per g [wet weight]), Bifidobacterium (log10 9.4 +/- 0.7 cells per g [wet weight]), and the Atopobium cluster (log10 9.3 +/- 0.7 cells per g [wet weight]). These five bacterial groups were detected in all 46 volunteers. Prevotella was found in only 46% of the subjects at a level of log10 9.7 +/- 0.8 cells per g (wet weight). Examination of changes in the population and the composition of the intestinal flora for six healthy adults over an 8-month period revealed that the composition of the flora of each volunteer remained stable throughout the test period.


Exposure-dependent misclassification of exposure in interaction analyses.
The objectives of this paper are to analyze the consequences of exposure misclassification on effect estimates in interaction analysis, and to develop a mathematical equation for the potentially biased estimate. The main point is to identify situations in which misclassification of the first exposure, dependent on the second exposure but independent on outcome status, leads to overestimation or underestimation of the interaction effect. We show that misclassification theoretically can cause overestimation of the interaction effect. Nevertheless, because the categories that yield overestimation due to misclassification are fewer than the categories that yield underestimation, and misclassification in reality mostly is multidimensional (more than one category are biased simultaneously), it is more likely that the effect of misclassification is underestimation rather than overestimation. Misclassification in the categories that lead to overestimation is compensated by misclassification in the categories that lead to underestimation. The magnitude of the biased estimate depends on the prevalences of the misclassified exposure, stratified for the second exposure and the outcome-the lower the prevalence, the smaller the bias.


Automated Identification of Diabetic Retinopathy Using Deep Learning.
Diabetic retinopathy (DR) is one of the leading causes of preventable blindness globally. Performing retinal screening examinations on all diabetic patients is an unmet need, and there are many undiagnosed and untreated cases of DR. The objective of this study was to develop robust diagnostic technology to automate DR screening. Referral of eyes with DR to an ophthalmologist for further evaluation and treatment would aid in reducing the rate of vision loss, enabling timely and accurate diagnoses. We developed and evaluated a data-driven deep learning algorithm as a novel diagnostic tool for automated DR detection. The algorithm processed color fundus images and classified them as healthy (no retinopathy) or having DR, identifying relevant cases for medical referral. A total of 75 137 publicly available fundus images from diabetic patients were used to train and test an artificial intelligence model to differentiate healthy fundi from those with DR. A panel of retinal specialists determined the ground truth for our data set before experimentation. We also tested our model using the public MESSIDOR 2 and E-Ophtha databases for external validation. Information learned in our automated method was visualized readily through an automatically generated abnormality heatmap, highlighting subregions within each input fundus image for further clinical review. We used area under the receiver operating characteristic curve (AUC) as a metric to measure the precision-recall trade-off of our algorithm, reporting associated sensitivity and specificity metrics on the receiver operating characteristic curve. Our model achieved a 0.97 AUC with a 94% and 98% sensitivity and specificity, respectively, on 5-fold cross-validation using our local data set. Testing against the independent MESSIDOR 2 and E-Ophtha databases achieved a 0.94 and 0.95 AUC score, respectively. A fully data-driven artificial intelligence-based grading algorithm can be used to screen fundus photographs obtained from diabetic patients and to identify, with high reliability, which cases should be referred to an ophthalmologist for further evaluation and treatment. The implementation of such an algorithm on a global basis could reduce drastically the rate of vision loss attributed to DR.


Antibody-selected mimics of hepatitis C virus hypervariable region 1 activate both primary and memory Th lymphocytes.
An ideal strategy that leads to a vaccine aimed at controlling viral escape may be that of preventing the replication of escape mutants by eliciting a T- and B-cell repertoire directed against many viral variants. The hypervariable region 1 (HVR1) of the putative envelope 2 protein that presents B and T epitopes shown to induce protective immunity against hepatitis C virus (HCV), might be suitable for this purpose if its immunogenicity can be improved by generating mimics that induce broad, highly cross-reactive, anti-HVR1 responses. Recently we described a successful approach to select HVR1 mimics (mimotopes) incorporating the variability found in a great number of viral variants. In this report we explore whether these mimotopes, designed to mimic B-cell epitopes, also mimic helper T-cell epitopes. The first interesting observation is that mimotopes selected for their reactivity to HVR1-specific antibodies of infected patients also do express HVR1 T-cell epitopes, suggesting that similar constraints govern HVR1-specific humoral and cellular immune responses. Moreover, some HVR1 mimotopes stimulate a multispecific CD4(+) T-cell repertoire that effectively cross-reacts with HVR1 native sequences. This may significantly limit effects as a T-cell receptor (TCR) antagonist frequently exerted by natural HVR1-variants on HVR1-specific T-cell responses. In conclusion, these data lend strong support to using HVR1 mimotopes in vaccines designed to prevent replication of escape mutants.


The BIOZO process--a biofilm system combined with ozonation: occurrence of xenobiotic organic micro-pollutants in and removal of polycyclic aromatic hydrocarbons and nitrogen from landfill leachate.
We present an assessment of xenobiotic organic micro-pollutants (XOM) occurrence and removal of polycyclic aromatic hydrocarbons (PAHs) in a novel biofilm system combined with ozonation, the BIOZO concept, treating partly stabilised landfill leachate. A novel, staged moving-bed biofilm reactor (SMBBR) design was implemented in laboratory- and pilot-scale, and the PAHs removal efficiency of controlled ozonation was assessed installing the ozonation step in the nitrate recirculation line (Position 1) or between the pre-anoxic and aerobic zones (Position 2). COD removal in a laboratory- and in a pilot-scale SMBBR system with and without ozonation is additionally addressed. Results obtained in a screening study (GC-ToF-MS) were used to compile a priority list of XOMs in leachate based on relative occurrence, showing PAHs as the predominant fraction. Biological treatment is shown to be an effective means to remove PAHs detected in the aqueous phase. PAH removal takes in most part place in the pre-anoxic zone, thereby decreasing toxicity exhibited by PAH on autotrophic nitrifier bacteria in the aerobic zone. Ozonation installed in Position 2 is shown to be superior over Position I in terms of COD, PAH and nitrogen removal efficiencies. We additionally demonstrate the potential of intermittent sludge ozonation as a means to decrease PAH concentrations in sludge wasted and to improve nitrogen removal in the BIOZO system.


Nanoporous Microsponge Particles (NMP) of Polysaccharides as Universal Carriers for Biomolecules Delivery.
Different polysaccharides-namely dextran, carboxymethyl dextran, alginate, and hyaluronic acid-were compared for the synthesis of nanoporous microsponges particles (NMPs) obtained from a one-pot self-precipitation/cross-linking process. The morphologies and sizes of the NMPs were evaluated comparatively with respect to polymer-to-polymer and cross-linker solvents (water-based vs. DMSO). We found that the radial distribution of the polymer in the near-spherical NMPs was found to peak either at the core or in the corona of the particle, depending both on the specific polymer or the solvent used for the formation of NMPs. The NMP porosity and the swelling capability were evaluated via scanning electron microscopy (SEM). The degradation study indicated that after 10 h incubation with a reducing agent, approximately 80% of the NMPs were disassembled into soluble polysaccharide chains. The adsorption and release capacity of each type of NMP were evaluated using fluorescently labeled bovine serum albumin and lysozyme as model proteins, highlighting a release time typically much longer than the corresponding adsorption time. The dependence of the adsorption-release performance on pH was demonstrated as well. Confocal microscopy images allowed us to probe the different distribution of labeled proteins inside the NMP. The safety and non-cytotoxicity of NMPs were evaluated after incubation with fibroblast 3T3 cells and showed that all types of NMPs did not adversely affect the cell viability for concentrations up to 2.25 μg/mL and an exposure time up to 120 h. Confocal microscopy imaging revealed also the effective interaction between NMPs and fibroblast 3T3 cells. Overall, this study describes a rapid, versatile, and facile approach for preparing a universal non-toxic, nanoporous carrier for protein delivery under physiological conditions.


Comparison of selective M3 and nonselective muscarinic receptor antagonists on gastrointestinal transit and bowel habits in humans.
Although in vitro studies show that muscarinic M(3) receptors primarily mediate the effects of acetylcholine on gastrointestinal contractility, the muscarinic receptor subtypes regulating gastrointestinal motor activity and transit in humans in vivo are unclear. We hypothesized that muscarinic M(3)-specific but not nonspecific receptor antagonists would delay gastrointestinal and colonic transit in humans. In this parallel-group study, gastric emptying, small intestinal transit, and colonic transit were assessed by scintigraphy on days 4-6 in 72 healthy subjects (49 women) who received placebo (n = 16), the M(3) antagonist darifenacin ER [7.5 mg (n = 20) or 15 mg daily (n = 17)], or the nonspecific antagonist tolterodine [4 mg daily (n = 19)] for 6 days. Bowel habits were recorded by daily diaries. Both doses of darifenacin substantially delayed [P < 0.01 vs. placebo (for both doses), P < 0.01 vs. tolterodine (for 15 mg)] small intestinal transit, i.e., colonic filling at 6 h (placebo [59.6 +/- 6.4%, mean +/- SE], 7.5 mg ER [34.4 +/- 6.1%], 15 mg ER [20.4 +/- 6.3%)]. Darifenacin (15 mg) also delayed (P < 0.01 vs. placebo and tolterodine) half-time for ascending colonic emptying [placebo (12.0 +/- 1.5 h), 7.5 mg (18.6 +/- 1.9 h), 15 mg (22.9 +/- 2.6 h)] and colonic transit (geometric center) at 24 [placebo (2.8 +/- 0.2), 7.5 mg (2.4 +/- 0.2), 15 mg (1.9 +/- 0.2)] but not 48 h. Darifenacin did not affect gastric emptying and tolterodine did not affect bowel habits or gastrointestinal transit. With muscarinic antagonists used at clinically approved doses, these findings demonstrate that muscarinic M(3) receptors regulate small intestinal and colonic transit in humans; colonic effects are more pronounced in the right than left colon. At doses that affect small and large intestinal transit, M(3) antagonists do not affect gastric emptying in humans. The efficacy of darifenacin in diarrhea-predominant irritable bowel syndrome should be evaluated.


Usefulness of combined PET/CT to assess regional lymph node involvement in gastric cancer.
The aim of this study was to evaluate the value of positron emission tomography/computed tomography (PET/CT) for preoperative staging of gastric cancer and to compare the diagnostic performance of PET/CT with that of contrast-enhanced computed tomography (CECT). We retrospectively reviewed 74 gastric cancer patients who underwent preoperative PET/CT and CECT, and subsequent curative surgical resection between April 2007 and July 2011. Preoperative PET/CT and CECT images for primary tumors of the stomach and lymph node metastases were reviewed retrospectively. The final diagnoses of primary tumors and LN metastases were based on histopathological specimens in all patients. Advanced gastric cancer was present in 65% of patients (n = 48), and the remaining patients had early gastric cancer (n = 26). Sixteen patients (22%) showed signet-ring-cell histology. For the detection of the primary tumor, the sensitivity of PET/CT was significantly higher than that of CECT (67% vs 55%, respectively; P = 0.049). For the evaluation of regional lymph node metastasis, the sensitivity, specificity, and accuracy of PET/CT and CECT were 34% and 51% (P = 0.065), 88% and 79% (P = 0.687), and 58% and 64% (P = 0.332), respectively. Neither PET/CT nor CECT detected regional lymph node metastases in early gastric cancer patients. Signet-ring-cell histology showed trends of non-FDG-avid lymph node metastases (odds ratio = 0.15, 95% confidence interval 0.17-1.37, P = 0.093). The accuracy of PET/CT is low and it is not a useful tool in the staging of gastric cancer overall in early gastric cancer and in signet-ring-cell carcinoma. Furthermore, the sensitivity of PET/CT could be inferior to that of CECT in the diagnosis of regional lymph node metastasis.


Fish-enhanced and soybean-enhanced supplemental snacks are acceptable among pregnant women in rural Kenya.
Although animal-source foods provide a rich source of complete protein and a variety of micronutrients, a majority of these foods are not accessible to a large proportion of populations in low-income nations. Locally available and affordable nutrient-dense dietary solutions that are accessible all year round can provide the most viable solution to improving food and nutrition security for these vulnerable populations. However, their potential to improve nutritional status among pregnant women has not been documented. The study was conducted to help guide the development of fish-enhanced and soybean-enhanced supplemental snacks and evaluate their acceptability among pregnant women in rural Kenya. We developed fish-enhanced and soybean-enhanced snacks using the small local lake fish omena (Rastrineobola argentea) and soybean flour. A cross-sectional study design was used to assess snack acceptance levels among HIV-infected and -noninfected pregnant women, two high-risk groups for nutrient deficiencies. Overall, 96%, 80%, and 81% of participants, indicated that they liked the taste, odor, texture, and color of wheat, fish-enhanced, or soybean-enhanced snacks, respectively. No significant differences were noted across participants' HIV status. Focus group discussions with the women further supported results from the quantitative ratings. Our analysis demonstrates the feasibility of developing acceptable, nutrient-dense food supplements using locally available foods in rural Kenya and contributes culturally acceptable, affordable, and sustainable solutions to the problem of undernutrition among pregnant women in low-income countries in sub-Saharan Africa.


Impact of a systematic education model for palliative care in cancer.
In clinical practice, pharmacists are continually required to improve their knowledge and expertise; however, the postgraduate education system for professional development cannot be confidently stated to be well established. The establishment of a systematic and multifaceted educational curriculum should be useful to improve home care and pharmacists' contribution; therefore, we developed a curriculum in collaboration with the university faculty of pharmaceutical sciences, department of pharmacy in hospital, and the Fukuoka Pharmaceutical Association. Class topics were extracted from the "Kanwa-Iryou-Yakugaku", edited by the Japanese Society for Pharmaceutical Palliative Care and Sciences. The items are necessary to perform palliative care as a pharmacist. A class schedule of 6 days (24 classes in total) was formulated. Questionnaires on comprehension degree before and after each class were provided to the participants. Comprehension was assessed on a scale of 1 to 10, where "I do not understand at all" was 1 and "I understand enough" was 10. The average recovery rates of questionnaires from each class were 92.6 % and 88.9 % before and after class, respectively. The average number of participants who completely answered the questionnaire before and after class was 45.6; therefore, these data were analyzed. Comprehension degree on each topic had significantly increased after attendance of all classes (p < 0.01). The comprehension degree of participants of the medical science of palliative care did greatly improve. Consequently, it is clear that the standard education model constructed was meaningful for the professional development of pharmacists in palliative care medicine.


Metabolic activity and monoclonal antibody production of Salmonella enteritidis O and H antigen specific hybridoma cells in static culture.
The aim of the present work was to study the kinetics of two hybridomas that produce monoclonal antibody against Salmonella Enteritidis O (5A8) and H (D7) antigen. The hybridomas originated from the Ag8x653 (5A8) and Sp2/O (D7) myeloma cell lines. The relationship between the uptake of glucose and glutamine and the release of the lactate and ammonia and monoclonal antibody production into hybridoma growth were investigated in static culture with serum-containing DMEM/F:12 medium for the determination of pilot-production strategies of the hybridomas. Results showed that glucose and glutamine concentrations were reduced, with an increase in ammonia and lactate concentration in the culture medium. The hybridoma cell line 5A8 has shown lower metabolic activities compared with D7, whereas its monoclonal antibody productivity was found to be two-fold higher than the D7. MAb production by the hybridoma cell line 5A8 seems promising, considering the moderate level of productivity compared to that found in the literature.


Perylo[1,12-b,c,d] Thiophene Tetraesters: A New Class of Luminescent Columnar Liquid Crystals.
Perylo[1,12-b,c,d] thiophene tetraesters exhibiting wide-range hexagonal columnar phase have been synthesized. These compounds also exhibit good homeotropic alignment in the liquid-crystalline phase which is very important for the device fabrication. These compounds showed sky-blue luminescence in solution under the long-wavelength UV light. With high solubility and high quantum yield these compounds can serve as standards to measure quantum yields of unknown samples. This new class of materials is promising, considering the emissive nature and stabilization of hexagonal columnar mesophase over a wide thermal range and ease of synthesis.


Hydrophobic mannosides act as acceptors for trypanosome alpha-mannosyltransferases.
A series of hydrophobic mannosides were synthesized and tested for their ability to act as acceptor substrates for mannosyltransferases in a Trypanosoma brucei cell-free system. The thiooctyl alpha-mannosides and octyl alpha-mannosides all accepted single mannose residues in alpha-linkage, as judged by thin layer chromatography of the products before and after jack bean alpha-mannosidase digestion. The mannosylation reactions were inhibited by amphomycin, suggesting that the immediate donor was dolichol-phosphate-mannose (Dol-P-Man) in all cases. The transferred alpha-mannose residues were shown to be both alpha 1-2 and alpha 1-6 linked by Aspergillus phoenicis alpha-mannosidase and acetolysis treatments, respectively. These data suggest that the compounds can act as acceptor substrates for the Dol-P-Man dependent alpha 1-2 and alpha 1-6 mannosyltransferases of the GPI biosynthetic pathway and/or the dolichol-cycle of protein N-glycosylation. One of the compounds, Man alpha 1-6 Man alpha 1-O-(CH2)7CH3, inhibited endogenous GPI biosynthesis in the cell-free system, suggesting that it could be a substrate for the trypanosome Dol-P-Man:Man2GlcN-Pl alpha 1-2 mannosyltransferase.


A proof of concept that experience-based management of endometriosis can complement evidence-based guidelines.
Management of endometriosis should be based on the best available evidence. The pyramid of evidence reflects unbiased observations analysed with traditional statistics. Evidence-based medicine (EBM) is the clinical interpretation of these data by experts. Unfortunately, traditional statistical inference can refute but cannot confirm a hypothesis and clinical experience is considered a personal opinion. A proof of concept to document clinical experience by considering each diagnosis and treatment as an experiment with an outcome, which is used to update subsequent management. Experience and knowledge-based questions were answered on a 0 to 10 visual analogue scale (VAS) by surgery-oriented clinicians with experience of > 50 surgeries for endometriosis. The answers reflect the collective clinical experience of managing >10.000 women with endometriosis. Experience-based management was overall comparable as approved by >75% of answers rated ≥ 8/10 VAS. Knowledge-based management was more variable, reflecting debated issues and differences between experts and non-experts. The collective experience-based management of those with endometriosis is similar for surgery-oriented clinicians. Results do not conflict with EBM and are a Bayesian prior, to be confirmed, refuted or updated by further observations. Collective experience-based management can be measured and is more than a personal opinion. This might extend EBM trial results to the entire population and add data difficult to obtain in RCTs, such as many aspects of surgery.


Discharge decision-making in a medical ICU: characteristics of unexpected readmissions.
To identify those patients most likely to be readmitted to a Medical ICU (MICU), we studied 512 MICU admissions during a 1-yr period. There were 36 readmissions within the same hospitalization in that interval. When ICU deaths and short-term drug overdoses were excluded, these 36 readmissions comprised 12% of all patients discharged from MICU. The mortality rate of this group was 58%, greater than twice the overall mortality rate for the year. Fifty-three % of the patients were readmitted because of recurrence of their initial disease on admission, with septicemia being the most common disease. Another 30% were readmitted because of a new complication, one-half of which were medication toxicities. We have attempted to delineate diseases, medications, and complications which may predict the high-risk discharge from MICU. Further studies to identify high-risk and low-risk admissions and discharges will effect better use of intensive care.


Critical role of monocyte chemoattractant protein-1 receptor CCR2 on monocytes in hypertension-induced vascular inflammation and remodeling.
Activated monocytes are present in the arterial walls of hypertensive patients and animals. Monocyte chemoattractant protein-1 (MCP-1), which controls monocyte function through its receptor (CCR2), is implicated in hypertensive inflammatory changes in the arterial wall. The role of CCR2 expression on monocytes in hypertension-induced vascular remodeling, however, has not been addressed. We hypothesized that CCR2 on monocytes is critical in hypertension-induced vascular inflammation and remodeling. Hypertension was induced by infusion of angiotensin II (Ang II) into wild-type mice, CCR2-deficient (CCR2-/-) mice, and bone marrow-transferred mice with a leukocyte-selective CCR2 deficiency (BMT-CCR2-/-). In wild-type mice, Ang II increased CCR2 intensity in circulating monocytes, which was prevented by an Ang II type-1 (AT1) receptor blocker or blunted in AT1 receptor-deficient mice. Enhanced CCR2 intensity on monocytes was observed in hypertensive patients and rats, and was reduced by treatment with the Ang II receptor blocker, supporting the clinical relevance of the observation in mice. In CCR2-/- and BMT-CCR2-/- mice, Ang II-induced vascular inflammation and vascular remodeling (aortic wall thickening and fibrosis) were blunted as compared with control mice. In contrast, Ang II-induced left ventricular hypertrophy developed in CCR2-/- and BMT-CCR2-/- mice. The present study suggests that CCR2 expression in monocytes has a critical role in vascular inflammation and remodeling in Ang II-induced hypertension, and possibly in other forms of hypertension.


Surgical approaches to obtaining optimal bowel function.
Approximately 50% of patients have an unsatisfactory functional result after traditional restorative rectal resection, and an even higher percentage, at least in the early postoperative period, suffers from urgency, frequent bowel movements, and occasional faecal incontinence. The rectal reservoir function is disturbed after restorative surgery. This is related to the size of the rectal remnant, the viscero-elastic properties, and the motility pattern of the neorectal wall, because segments of the remaining colon can only substitute for the rectum to a limited extent. A straight anastomosis is recommended when the rectal remnant (measured from the anal verge) is at least 7 to 8 cm. The side-to-end anastomosis is probably preferable to the end-to-end anastomosis. In contrast, a straight anastomosis at the levator plane cannot be recommended. If straight anastomosis is still considered, the descending colon should be used rather than the sigmoid colon. The colonic pouch was introduced to increase the neorectal volume and eliminate some of the functional disturbance associated with the reduced neorectal volume occurring after a straight colo-anal anastomosis. To obtain optimal functional results soon after surgery, a pouch should be used when the anastomosis is located 3 to 5 cm from the anal verge. The size of the pouch should not be too small. A staple line of 6 to 7 cm is a fair compromise between the low anterior resection syndrome and problems with evacuation. Since the descending colon has a thinner wall and often is healthier than the sigmoid colon, it should be the first choice for the anastomosis.


Exploring the efficacy and safety of a novel standardized ashwagandha (Withania somnifera) root extract (Witholytin®) in adults experiencing high stress and fatigue in a randomized, double-blind, placebo-controlled trial.
Stress is a state of homeostasis in the body being challenged, resulting in a systemic response. It has become more prevalent in recent years and affects mental and physical health. Evaluate the effects of ashwagandha on stress, fatigue, and sex hormones in overweight or mildly obese men and women with self-reported stress and fatigue. Two-arm, parallel-group, 12-week, randomized, double-blind, placebo-controlled trial on overweight or mildly obese men and women aged 40-75 years, supplementing with 200 mg of an ashwagandha root extract (Witholytin®) twice daily. Supplementation with ashwagandha was associated with a significant reduction in stress levels based on the Perceived Stress Scale (primary outcome); however, the improvements were not significantly different to the placebo group (p = 0.867). Based on the Chalder Fatigue Scale, there was a statistically significant reduction in fatigue symptoms in the ashwagandha group compared to the placebo group (p = 0.016), and participants taking ashwagandha also experienced a significant increase in heart rate variability (p = 0.003). However, there were no significant between-group differences in other self-report outcome measures. In the men taking ashwagandha, there was a significant increase in the blood concentrations of free testosterone (p = 0.048) and luteinizing hormone (p = 0.002) compared to the placebo group. The results of this study suggest that in overweight middle-to-older age adults experiencing high stress and fatigue, compared to the placebo, ashwagandha did not have a significantly greater impact on perceived stress levels. However, based on secondary outcome measures, it may have anti-fatigue effects. This may be via its impact on the autonomic nervous system. However, further research is required to expand on these current findings.


[Effect of immersion hypokinesia on the characteristics of programmed-type voluntary movements].
The effect of immersion hypokinesia on the precision of programme-type voluntary movements was investigated using standard test movements and quantitative analytical methods. The exposure did not cause disorders in the programme mechanisms but reduced significantly the precision range of the motor control system. The loss of precision was at its maximum (by 100% and over) on immersion day 3. The universality and consistency of the above changes indicated their close association with the specific exposure while the fast rate of their development suggested their reflex nature.


[Stratification of arrhythmia risk in infarction: reflexions in the light of large trials].
The identification of patients at higher risk of life-threatening ventricular arrhythmias after myocardial infarction still represents a clinically relevant problem, particularly after results of recent studies which support the efficacy of implantable cardioverter defibrillator (ICD) in reducing total mortality in patients with a previous myocardial infarction and left ventricular dysfunction, with and without additional risk markers. However, owing to the high cost of ICD therapy, an effective arrhythmic risk stratification may be desirable. The low diagnostic accuracy reported by various studies using single risk stratifiers (either invasive and non invasive) suggested a combined use of multiple parameters in order to improve the predictive power of the risk stratification algorithms. This approach, that takes into account the multifactorial genesis of malignant ventricular arrhythmias, has been demonstrated to be able to identify subgroups of patients at very high arrhythmic risk. In particular, a two-level algorithm based upon the selection of candidates to electrophysiologic study among patients with abnormal non-invasive testing, showed itself as a particularly effective tool for identification of such patients. In this paper the Authors summarize most recent results on the risk stratification protocols and the use of ICDs and provide an operative algorithm that keeps into account either aggressive and moderate approaches to patients surviving a myocardial infarction.


Human papillomavirus type 16 DNA from a vulvar carcinoma in situ is present as head-to-tail dimeric episomes with a deletion in the non-coding region.
A number of genital cancer biopsy samples were screened for the presence of human papillomavirus type 16 (HPV-16) DNA sequences. One of these samples (a vulvar carcinoma in situ) was found to contain more than 100 copies of HPV-16 DNA sequences per cell. Using this tumour DNA, a genomic library was constructed in bacteriophage lambda and the library was screened for recombinant phage containing HPV-16 sequences. Five recombinant phage clones were isolated and their DNA was analysed by restriction endonuclease digestion and blot hybridization. All five recombinants contained two copies of the HPV-16 genome present in a head-to-tail arrangement. The data are consistent with the presence of HPV-16 sequences in the tumour DNA arranged as genomic dimers in a circular episomal configuration. The HPV-16 genomes contained a deletion within the non-coding region, a region which includes the viral origin of DNA replication and transcriptional control sequences. Possible consequences of this deletion for viral replication and transcription are discussed.


The Association Between Risk Factors for Metabolic Syndrome and Meibomian Gland Disease in a Dry Eye Cohort.
Risk factors for metabolic syndrome include abdominal obesity, insulin resistance, hypertension, high triglycerides and/or low high-density lipoprotein cholesterol, and hyperglycemia. Risk factors for metabolic syndrome have been associated with dry eye disease; however, their association with meibomian gland disease (MGD), a subtype of dry eye, is unclear. In the present study, we investigated risk factors for metabolic syndrome in a dry eye cohort with and without MGD. This retrospective case-control study evaluated electronic medical records at a major urban outpatient medical center to identify patients with a known diagnosis of dry eye disease with and without MGD. Males and females were matched for age, smoking status, race, ethnicity, and body mass index (BMI). Patient demographics, anthropometric measurements, medical history, clinical findings, and serologies were analyzed. A diagnosis of MGD was based on clinical signs noted in the medical record. MGD was not associated with BMI, smoking, type 2 diabetes mellitus, hypertension or hyperlipidemia in this dry eye cohort. MGD was associated with male sex and increasing age. While increasing age was weakly correlated with decreased low density lipoprotein cholesterol and non-high density lipoprotein cholesterol, serum lipid levels were not associated with MGD. Importantly, we found that risk factors for metabolic syndrome are not specifically associated with an increase in MGD when compared to non-MGD dry eye subjects. While risk factors for metabolic syndrome are associated with dry eye disease, they likely reflect a chronic systemic state of low-grade inflammation that negatively impacts the function of both lacrimal and meibomian glands.


Introducing High School Students to Careers in Osteopathic Medicine.
An aging population and expansion of health care coverage under the Patient Protection and Affordable Care Act have led to a predicted deficit of primary care physicians by 2025. In response, medical schools must develop new strategies to identify students early in their educational pathways to encourage exploration of careers in science, technology, engineering, and mathematics (STEM) fields, especially medicine. Oklahoma State University Center for Health Sciences developed 2 internship programs, Oklahoma Science Training and Research Students (OKStars) and Native OKStars, to introduce high school students to osteopathic medicine. Native OKStars was designed to encourage Native American students, who are underrepresented in STEM fields, to pursue osteopathic medicine. These programs provided students with a 6-week immersion in biomedical research, along with weekly discussion groups with mentors. Participant questionnaire responses suggested that these programs were effective in introducing them to careers in osteopathic medicine as well as other STEM fields.


Validation of the automated method VIENA: an accurate, precise, and robust measure of ventricular enlargement.
In many retrospective studies and large clinical trials, high-resolution, good-contrast 3DT1 images are unavailable, hampering detailed analysis of brain atrophy. Ventricular enlargement then provides a sensitive indirect measure of ongoing central brain atrophy. Validated automated methods are required that can reliably measure ventricular enlargement and are robust across magnetic resonance (MR) image types. To validate the automated method VIENA for measuring the percentage ventricular volume change (PVVC) between two scans. Accuracy was assessed using four image types, acquired in 15 elderly patients (five with Alzheimer's disease, five with mild cognitive impairment, and five cognitively normal elderly) and 58 patients with multiple sclerosis (MS), by comparing PVVC values from VIENA to manual outlining. Precision was assessed from data with three imaging time points per MS patient, by measuring the difference between the direct (one-step) and indirect (two-step) measurement of ventricular volume change between the first and last time points. The stringent concordance correlation coefficient (CCC) was used to quantify absolute agreement. CCC of VIENA with manual measurement was 0.84, indicating good absolute agreement. The median absolute difference between two-step and one-step measurement with VIENA was 1.01%, while CCC was 0.98. Neither initial ventricular volume nor ventricular volume change affected performance of the method. VIENA has good accuracy and good precision across four image types. VIENA therefore provides a useful fully automated method for measuring ventricular volume change in large datasets. VIENA is a robust, accurate, and precise method for measuring ventricular volume change.


Establishing cross-discipline consensus on contraception, pregnancy and breast feeding-related educational messages and clinical practices to support women with rheumatoid arthritis: an Australian Delphi study.
Recognising the need for a best-practice and consistent approach in providing care to women with rheumatoid arthritis (RA) in relation to (1) general health, (2) contraception, (3) conception and pregnancy, (4) breast feeding and (5) early parenting, we sought to achieve cross-discipline, clinical consensus on key messages and clinical practice behaviours in these 5 areas. 3-round eDelphi study. In round 1, panellists provided free-text responses to open-ended questions about care for women with RA across the 5 areas. Subsequently, panellists refined and scored the synthesised responses, presented as metathemes, themes and detailed elements. Where ≥5% of panellists did not support a theme in a given round, it was removed. Panel of practicing Australian rheumatologists (n=22), obstetricians/obstetric medicine physicians (n=9) and pharmacists (n=5). 34 (94.4%) panellists participated in all 3 rounds. The panel supported 18 themes across the 5 areas (support/strongly support: 88.2-100%) underpinned by 5 metathemes. Metathemes focused on coordination in information delivery, the mode and timing of information delivery, evidence underpinning information, engagement of the right health professionals at the right time and a non-judgemental approach to infant feeding. Themes included practices for primary prevention of chronic disease and their sequelae, the importance of contraception and planning pregnancy and breast feeding, close monitoring of medications, supporting mental well-being, managing disease activity and providing practical support for early parenting. A cross-disciplinary clinical panel highly supported key information and clinical practices in the care for women with RA across the continuum of contraception to early parenting within a whole-person, chronic disease management approach.


Molecular mechanisms of resveratrol (3,4,5-trihydroxy-trans-stilbene) and its interaction with TNF-related apoptosis inducing ligand (TRAIL) in androgen-insensitive prostate cancer cells.
Although resveratrol, an active ingredient derived from grapes and red wine, possesses chemopreventive properties against several cancers, the molecular mechanisms by which it inhibits cell growth and induces apoptosis have not been clearly understood. Here, we examined the molecular mechanisms of resveratrol and its interactive effects with TRAIL on apoptosis in prostate cancer PC-3 and DU-145 cells. Resveratrol inhibited cell viability and colony formation, and induced apoptosis in prostate cancer cells. Resveratrol downregulated the expression of Bcl-2, Bcl-X(L) and survivin and upregulated the expression of Bax, Bak, PUMA, Noxa, and Bim, and death receptors (TRAIL-R1/DR4 and TRAIL-R2/DR5). Treatment of prostate cancer cells with resveratrol resulted in generation of reactive oxygen species (ROS), translocation of Bax to mitochondria and subsequent drop in mitochondrial membrane potential, release of mitochondrial proteins (cytochrome c, Smac/DIABLO, and AIF) to cytosol, activation of effector caspase-3 and caspase-9, and induction of apoptosis. Resveratrol-induced ROS production, caspase-3 activity and apoptosis were inhibited by N-acetylcysteine. Bax was a major proapoptotic gene mediating the effects of resveratrol as Bax siRNA inhibited resveratrol-induced apoptosis. Resveratrol enhanced the apoptosis-inducing potential of TRAIL, and these effects were inhibited by either dominant negative FADD or caspase-8 siRNA. The combination of resveratrol and TRAIL enhanced the mitochondrial dysfunctions during apoptosis. These properties of resveratrol strongly suggest that it could be used either alone or in combination with TRAIL for the prevention and/or treatment of prostate cancer.


Treatment of acromegaly in the era of personalized and predictive medicine.
Acromegaly is a chronic disorder usually diagnosed late in the disease evolution. Such delayed diagnosis, together with the inability to achieve the treatment goals of normalizing biochemical disease markers and controlling tumour mass may result in substantial morbidity and mortality. Somatostatin analogues (SSA) are accepted as first-line medical therapy or as second-line therapy in patients undergoing unsuccessful surgery and are considered a cornerstone in the treatment of acromegaly. However, because a high percentage of patients experience SSA medical treatment failure, the identification of biomarkers associated with a successful or unsuccessful response to all classes of medical therapy would help in the choice of treatment and potentially allow for a quicker normalization of biochemical parameters. The current treatment algorithms for acromegaly are based upon a "trial-and-error" approach with additional treatment options provided when disease is not controlled. In many other diseases, therapeutic algorithms have been evolving towards personalized treatment with the medication that best matches individual disease characteristics, using biomarkers that identify therapeutic response. Additionally, a personalized approach to complementary treatment of comorbidities present in the acromegalic patient is also required. This review will discuss the development of a potential treatment algorithm for acromegaly addressing the biochemical control of the disease as well of its associated comorbidities, under a personalized approach based upon markers of prognostic and predictive significance, such as tumour size, MRI adenoma signal, GH value after acute octreotide test, granular adenoma pattern, Ki-67, somatostatin receptor phenotype, aryl hydrocarbon-interacting protein expression, gsp mutations, RAF kinase activity, E-cadherin and beta-arrestin-1.


Whole-body vibration perception thresholds of recumbent subjects--Part 1: Supine posture.
The objective of the present study was to determine perception thresholds for whole-body vibration in the recumbent supine posture in vertical and horizontal directions and to compare the present results with ones obtained at the same laboratory in 1984 and also with ISO frequency weighting curves both in ISO 2631-1 and in ISO 2631-2. The frequency characteristic and thresholds values of the present results are similar to previous results. Weighting curves of ISO, however, show quite different characteristic in both vertical and horizontal directions compared to the present characteristics. It is not suitable to apply weighting curves Wk for the vertical direction, Wd for the horizontal direction and combined curves in ISO to recumbent posture from the results of this present study.


Pre-incubation with human umbilical cord derived mesenchymal stem cells-exosomes prevents cisplatin-induced renal tubular epithelial cell injury.
The administration of cisplatin is limited due to its nephrotoxicity, and prevention of this nephrotoxicity of cisplatin is difficult. Mesenchymal stem cell (MSC)-derived exosomes have been implicated as a novel therapeutic approach for tissue injury. In vitro, the NRK cells pre-incubated with HUMSC-exosomes increased the Cp-inhibited cell viability, proliferation activity, and the cell proportion in G1-phase and inhibited Cp-induced cell apoptosis. Furthermore, the expression levels of apoptotic marker proteins Bim, Bad, Bax, cleaved caspase-3, and cleaved caspase-9 induced by Cp in the NRK cells were decreased by pre-incubating with HUMSC-exosomes. Our findings indicated that the exosomes from HUMSCs can effectively increase the survival rate and inhibit cell apoptosis of NRK cells. Therefore, pre-treatment of HUMSC-exosomes may be a new method to improve the therapeutic effect of cisplatin. Exosomes were isolated from human umbilical cord derived mesenchymal stem cells (HUMSCs). Co-culture of normal rat renal tubular epithelial cells (NRK) and the absorption of exogenous exosomes by NRK cells were examined in vitro. Then the NRK cells were incubated with exosomes from HUMSCs and cisplatin (Cp). Cells were harvested for MTT assay, cloning formation, flow cytometry, and Western blot.


Decreased expression of the DNA mismatch repair gene Mlh1 under hypoxic stress in mammalian cells.
The hypoxic tumor microenvironment has been shown to contribute to genetic instability. As one possible mechanism for this effect, we report that expression of the DNA mismatch repair (MMR) gene Mlh1 is specifically reduced in mammalian cells under hypoxia, whereas expression of other MMR genes, including Msh2, Msh6, and Pms2, is not altered at the mRNA level. However, levels of the PMS2 protein are reduced, consistent with destabilization of PMS2 in the absence of its heterodimer partner, MLH1. The hypoxia-induced reduction in Mlh1 mRNA was prevented by the histone deacetylase inhibitor trichostatin A, suggesting that hypoxia causes decreased Mlh1 transcription via histone deacetylation. In addition, treatment of cells with the iron chelator desferrioxamine also reduced MLH1 and PMS2 levels, in keeping with low oxygen tension being the stress signal that provokes the altered MMR gene expression. Functional MMR deficiency under hypoxia was detected as induced instability of a (CA)(29) dinucleotide repeat and by increased mutagenesis in a chromosomal reporter gene. These results identify a potential new pathway of genetic instability in cancer: hypoxia-induced reduction in the expression of key MMR proteins. In addition, this stress-induced genetic instability may represent a conceptual parallel to the pathway of stationary-phase mutagenesis seen in bacteria.


Design and rationale of the atrial fibrillation occurring transiently with stress (AFOTS) follow-up cohort study.
Atrial fibrillation occurring transiently with stress (AFOTS) describes the first detection of AF in a patient who is hospitalized for a non-cardiac medical illness or following non-cardiac surgery. Uncertainty exists whether episodes of AFOTS are due to reversible precipitants and will not recur after recovery, or if they are paroxysmal atrial fibrillation (AF) that is detected during inpatient cardiac monitoring. Previous studies have used retrospective, non-systematic and ultimately low-sensitivity protocols to investigate the recurrence of AF in patients with AFOTS. The prospective, multi-center, investigator-initiated AFOTS Follow-Up Cohort Study will enroll 138 case patients with AFOTS in the setting of non-cardiac surgery or medical illness, matched 1:1 with control patients for age, sex, stressor, and hospital unit. Participants will wear a 14-day ECG heart monitor at 1 and 6 months after hospital discharge. Over 12 months of follow-up, we will collect data regarding participant's medications, and clinical events. The primary endpoint is detection of 30 or more seconds of AF after hospital discharge. To date, 50% of the target sample has been enrolled. The study is expected to complete enrolment in mid-2019 and conclude 1 year later. The AFOTS follow-up study will employ a systematic protocol to detect AF and will provide a precise and valid estimate of AF recurrence following AFOTS. This study will establish whether patients with AFOTS have an increased propensity to AF after hospitalization as compared to matched controls and may inform the management of this population.


Thinking About Development: The Value of Animal-Based Research for the Study of Human Development.
Gottlieb promoted the value of a developmental psychobiological systems approach to the study of human development. This approach recognizes the importance of comparative, animal-based research to advancing our understanding of the complexities and dynamics of the process of development. The major contribution of animal developmental studies is their provision of food for thought (hypotheses, not facts) about human development and general principles of development. Here we briefly describe how, guided by Gottlieb's pioneering vision, we have utilized coordinated studies of non-human animal and human infants to begin to identify patterns of selective attention and perceptual processing that are common across species in early development. Our converging findings highlight the importance of multimodal (intersensory) redundancy in guiding and constraining early perceptual learning in avian and mammalian species.


Antidepressants in bipolar disorder: the case for caution.
The 2002 American Psychiatric Association (APA) guidelines for the treatment of bipolar disorder recommended more conservative use of antidepressants. This change in comparison with previous APA guidelines has been criticized, especially from some groups in Europe. The Munich group in particular has published a critique of assumptions underlying the conservative recommendations of the recent APA treatment guidelines. In this paper, we re-examine the argument put forward by the Munich group, and we demonstrate that indeed, conceptually and empirically, there is a strong rationale for a cautious approach to antidepressant use in bipolar disorder, consistent with, and perhaps even more strongly than, the APA guidelines. This rationale is based on support for the following four propositions: (i) The risk of antidepressant induced mood-cycling is high, (ii) Antidepressants have not been shown to definitively prevent completed suicides and reduce mortality, whereas lithium has, (iii) Antidepressants have not been shown to be more effective than mood stabilizers in acute bipolar depression and have been shown to be less effective than mood stabilizers in preventing depressive relapse in bipolar disorder and (iv) Mood stabilizers, especially lithium and lamotrigine, have been shown to be effective in acute and prophylactic treatment of bipolar depressive episodes. We therefore draw three conclusions from this interpretation of the evidence: (i) There are significant risks of mania and long-term worsening of bipolar illness with antidepressants, (ii) Antidepressants should generally be reserved for severe cases of acute bipolar depression and not routinely used in mild to moderate cases and (iii) Antidepressants should be discontinued after recovery from the depressive episode, and maintained only in those who repeatedly relapse after antidepressant discontinuation (a minority we judge to represent only about 15-20% of bipolar depressed patients).


Mechanisms of tolerance and high degradation capacity of the herbicide mesotrione by Escherichia coli strain DH5-α.
The intensive use of agrochemicals has played an important role in increasing agricultural production. One of the impacts of agrochemical use has been changes in population structure of soil microbiota. The aim of this work was to analyze the adaptive strategies that bacteria use to overcome oxidative stress caused by mesotrione, which inhibits 4-hydroxyphenylpyruvate dioxygenase. We also examined antioxidative stress systems, saturation changes of lipid membranes, and the capacity of bacteria to degrade mesotrione. Escherichia coli DH5-á was chosen as a non-environmental strain, which is already a model bacterium for studying metabolism and adaptation. The results showed that this bacterium was able to tolerate high doses of the herbicide (10× field rate), and completely degraded mesotrione after 3 h of exposure, as determined by a High Performance Liquid Chromatography. Growth rates in the presence of mesotrione were lower than in the control, prior to the period of degradation, showing toxic effects of this herbicide on bacterial cells. Changes in the saturation of the membrane lipids reduced the damage caused by reactive oxygen species and possibly hindered the entry of xenobiotics in the cell, while activating glutathione-S-transferase enzyme in the antioxidant system and in the metabolizing process of the herbicide. Considering that E. coli DH5-α is a non-environmental strain and it had no previous contact with mesotrione, the defense system found in this strain could be considered non-specific. This bacterium system response may be a general adaptation mechanism by which bacterial strains resist to damage from the presence of herbicides in agricultural soils.


Should foliar cadmium concentrations be expressed on a dry weight or dry ash weight basis?
Foliar analysis is a valuable tool for evaluating the pollution status of forests. However, the use of foliar diagnosis in large-scale surveys is a complicated process owing to the high variability within the crown. The method used to express foliar concentrations has often been found to diminish the variability. The effect of the method used to express element concentrations on the spatial variability of cadmium (Cd) in the leaves of crack willow (Salix fragilis L.) was investigated by sampling the leaves of one willow at 292 locations in the crown, each sampling location having a volume of 0.027 m3 (0.3 m x 0.3 m x 0.3 m). Cadmium showed a distinct spatial trend in the crown of the tree. Concentrations as low as 2.4 mg kg(-1) dry weight (DW) or 23.1 mg kg(-1) dry ash weight (DAW) were obtained in the top of the crown, and 10.6 mg kg(-1) DW or 73.0 mg kg(-1) DAW in the bottom of the crown. The lower relative standard deviation and weaker correlation with the sampling height support the use of DAW in large-scale surveys especially. The lower variability of the DAW Cd concentration makes this variable less sensitive to fluctuations caused by differences in growing conditions and sampling methodology. However, the majority of publications in this field report metal concentrations on a DW basis. Therefore, the restrictions set on the use of results expressed on a DAW basis in large-scale surveys of foliar metal concentrations have to be offset against the advantages offered by a reduction of the variability in metal concentrations.


Effects of ischemic preconditioning on contractile and metabolic function during hypoperfusion in dogs.
We examined the effects of ischemic preconditioning (i.p.) on metabolic and contractile function during coronary hypoperfusion in dogs. After the left anterior descending coronary artery (LAD) was occluded for 5 min (i.p.) and reperfused for 10 min, coronary blood flow (CBF) of the LAD was decreased to 33% of the control. I.p. increased (P < 0.05) lactate extraction ratio and the pH of coronary venous blood and decreased (P < 0.05) myocardial oxygen consumption and fractional shortening during hypoperfusion compared with those in the control group, although i.p. did not change the endocardial-to-epicardial blood flow ratio of the regional myocardium during hypoperfusion. I.p. increased (P < 0.05) the adenosine levels in coronary venous blood during hypoperfusion. I.p. increased (P < 0.05) myocardial ecto-5'-nucleotidase activity. Administration of 8-sulfophenyltheophylline or alpha, beta-methyleneadenosine 5'-diphosphate blunted the i.p.-induced changes in metabolic and contractile parameters during hypoperfusion. These results suggest that i.p. reduced the severity of anaerobic myocardial metabolism of ischemic hearts by increasing the adenosine levels via an extracellular pathway.


Analysis of recognition elements for mitochondrial processing peptidase using artificial amino acids: roles of the intervening portion and proximal arginine.
We recently demonstrated, using synthetic peptides modeled on the extension peptide of malate dehydrogenase, that amino acid residues present at the proximal and distal positions relative to the cleavage site are critical determinants for the recognition of substrates by mitochondrial processing peptidase [Niidome et al. (1994) J. Biol. Chem. 269, 24719-24722). While the proximal arginine is unexceptionally located at the -2 position, the position of the distal residue varies among mitochondrial precursor proteins. Between the proximal and distal residues, proline and/or glycine are present in most mitochondrial precursor proteins, and they are considered to play a role in the specific recognition of a substrate by the peptidase. To elucidate the role of the intervening portion, we introduced a non-natural amino acid [2-(2-aminoethoxy)acetic acid] between the distal and proximal residues. We also analyzed the functional elements in the proximal arginine by replacing the residue with various arginine or lysine analogs. The results of kinetic studies indicated that the intervening portion should be flexible for efficient processing, and that the guanidino group of the proximal arginine is recognized by the peptidase through hydrogen and ionic bonds.


Intrauterine devices & infection: review of the literature.
The relationship between use of an intrauterine device (IUD) and pelvic inflammatory disease (PID) has been studied extensively over the past 50 years. Previous research has led to considerable controversy and debate. Numerous limitations in the studies make it difficult to draw any firm conclusions from the past research or to design new approaches to study the topic. The main research barriers include uncertainty of infection/diagnoses, and inappropriate comparison groups for IUD users. Natural history studies of the aetiology of disease and observational research among IUD users suggest that the risk of PID is very low. Research linking previous IUD use to the more distant endpoint of tubal infertility reveals that the risks may be even lower than the risks of PID.


Stimulation of normal inducer T cell clones with antigen presented by purified Ia molecules in planar lipid membranes: specific induction of a long-lived state of proliferative nonresponsiveness.
Culture of normal inducer T cell clones with antigen and purified Ek beta:Ek alpha incorporated into planar lipid membranes resulted in specific T cell activation as determined by cell volume increase and IL 3 production. However, in contrast to results obtained with T cell hybridomas, antigen presentation by planar membranes did not induce measurable IL 2 production, and proliferative responses were not detected. Rather, recognition of only Ek beta:Ek alpha and antigen resulted in the specific induction of a long-lived state of proliferative nonresponsiveness to subsequent stimulation by conventional APC and antigen. Induction of nonresponsiveness required protein synthesis, and was not simply due to the absence of IL 2. The antigen-nonresponsive cells could respond to either PMA plus ionomycin or IL 2, and they expressed normal levels of surface antigen-receptor molecules. These results demonstrate that recognition by normal T cell clones of antigen and Ia molecules in the absence of other accessory cell molecules and signals results in a prolonged state of proliferative nonresponsiveness, possibly similar to a state of T cell tolerance in vivo.


Robustness of Light-Transport Processes to Bending Deformations in Graded-Index Multimode Waveguides.
Light transport through a multimode optical waveguide undergoes changes when subjected to bending deformations. We show that optical waveguides with a perfectly parabolic refractive index profile are almost immune to bending, conserving the structure of propagation-invariant modes. Moreover, we show that changes to the transmission matrix of parabolic-index fibers due to bending can be expressed with only two free parameters, regardless of how complex a particular deformation is. We provide detailed analysis of experimentally measured transmission matrices of a commercially available graded-index fiber as well as a gradient-index rod lens featuring a very faithful parabolic refractive index profile. Although parabolic-index fibers with a sufficiently precise refractive index profile are not within our reach, we show that imaging performance with standard commercially available graded-index fibers is significantly less influenced by bending deformations than step-index types under the same conditions. Our work thus predicts that the availability of ultraprecise parabolic-index fibers will make endoscopic applications with flexible probes feasible and free from extremely elaborate computational challenges.


Increased overt attention to objects in early deaf adults: An eye-tracking study of complex naturalistic scenes.
The study of selective attention in people with profound deafness has repeatedly documented enhanced attention to the peripheral regions of the visual field compared to hearing controls. This finding emerged from covert attention studies (i.e., without eye-movements) involving extremely simplified visual scenes and comprising few visual items. In this study, we aimed to test whether this key finding extends also to overt attention, using a more ecologically valid experimental context in which complex naturalistic images were presented for 3 s. In Experiment 1 (N = 35), all images contained a single central object superimposed on a congruent naturalistic background (e.g., a tiger in the woods). At the end of the visual exploration phase, an incidental memory task probed the participants' recollection of the seen central objects and image backgrounds. Results showed that hearing controls explored and remembered the image backgrounds more than deaf participants, who lingered on the central object to a greater extent. In Experiment 2 we aimed to disentangle if this behaviour of deaf participants reflected a bias in overt space-based attention towards the centre of the image, or instead, enhanced object-centred attention. We tested new participants (N = 42) in the visual exploration task adding images with lateralized objects, as well as images with multiple object or images without any object. Results confirmed increased exploration of objects in deaf participants. Taken together our novel findings show limitations of the well-known peripheral attention bias of deaf people and suggest that visual object-centred attention may also change after prolonged auditory deprivation.


Peptides for functionalization of InP semiconductors.
The challenge is to achieve high specificity in molecular sensing by proper functionalization of micro/nano-structured semiconductors by peptides that reveal specific recognition for these structures. Here we report on surface modification of the InP semiconductors by adhesion peptides produced by the phage display technique. An M13 bacteriophage library has been used to screen 10(10) different peptides against the InP(001) and the InP(111) surfaces to finally isolate specific peptides for each orientation of the InP. MALDI-TOF/TOF mass spectrometry has been employed to study real affinity of the peptide towards the InP surfaces. The peptides serve for controlled placement of biotin onto InP to bind then streptavidin. Our Atomic Force Microscopy study revealed a total surface coverage of molecules when the InP surface was functionalized by its specific biotinylated peptide (YAIKGPSHFRPS). Finally, fluorescence microscopy has been employed to demonstrate the preferential attachment of the peptide onto a micro-patterned InP surface. Use of substrate specific peptides could present an alternative solution for the problems encountered in the actually existing sensing methods and molecular self-assembly due to the unwanted unspecific interactions.


ApoB-containing lipoproteins promote infectivity of chlamydial species in human hepatoma cell line.
To evaluate the direct binding of two main chlamydial biovars (C. trachomatis and C. pneumoniae) to plasma lipoproteins and its effect on chlamydial infection rate in human hepatoma cell line (HepG2 cells). Murine plasma lipoproteins were fractionated and isolated using fast-performance liquid chromatography (FPLC), spotted on nitrocellulose membrane and incubated with chlamydial suspensions. Direct binding of chlamydial particles to lipoprotein fractions has been studied using lipopolysaccharide-specific antibodies in immuno-dot blot binding assay and immunoprecipitation analysis. Immunostaining protocol as well as flow cytometry analysis have been employed to study the infectivity rate of chlamydial species in HepG2 cells. Elementary bodies of both C. trachomatis and C. pneumoniae bind ApoB-containing fractions of plasma lipoproteins. That binding becomes stronger when heat-denatured FPLC fractions are used, suggesting a primary role of apolipoproteins in interaction between chlamydial particle and lipoprotein. Both chlamydial biovars efficiently propagate in human hepatoma cell line - HepG2 cells even in serum free conditions forming late-stage inclusion bodies and releasing extracellular elementary bodies. Preincubation of C. trachomatis and C. pneumoniae with native ApoB-containing lipoproteins enhances the rate of chlamydial infection in HepG2 cells. A productive infection caused by C. trachomatis and C. pneumoniae may take place in human-derived hepatocytes revealing hepatic cells as possible target in chlamydial infection. Obtained results may suggest the participation of lipoprotein receptors in the mechanism of attachment and/or entry of chlamydial particles into target cells.


[Radiation therapy for uterine cervix cancer: importance of evaluation of pre-treatment tumor size with MR imaging].
From May 1992 through December 1995, a total of 42 patients with previously untreated squamous cell carcinoma of the uterine cervix were treated by using middle-dose-rate intracavitary therapy, and their previously treated local tumor volume was evaluated with MRI. According to the staging of FIGO, 2 patients were classified as Stage IB, 2 as IIA, 18 as IIB, 1 as IIIA, 14 as IIIB, 2 as IVA and 3 as IVB. Cumulative 3-year survival rates were 89% in Stage IIB and 54% in IIIB. 3-year local control rates were 100% in Stage IIB and 67% in IIIB. On the other hand, the cumulative survival rate for the local control group in all stages was 100% at 2 years and 82% at 3 years. For the pelvic failure group it was 41% at 2 years. All of those in the pelvic failure group had bulky local tumor sizes of more than 60 mm in the previous treatment MRI study. The 3-year cumulative survival rate and local control rate for tumors less than 60 mm were 84% and 100%, but for tumors over 60 mm the respective rates were 45% and 33% (p < 0.01). The a results showed that the group with pretreatment of local tumors over 60 mm on MRI had a poor outcome. We concluded that pretreatment tumor volume is an important factor in prognosis and the evaluation of pretreatment tumor volume by imaging study (MRI) is necessary.


Interaction of cholesterol with photoactivable phospholipids in sonicated vesicles.
Photoactivable phospholipids containing either alpha-diazo-beta-trifluoropropionyloxy or m-diazirinophenoxyl groups in the omega-positions of sn-2 fatty acyl chains were synthesized and incorporated into sonicated vesicles containing 33 mol% of cholesterol. Photolysis of the vesicles at 350 nm produced covalent cross-links between the synthetic phospholipids and cholesterol. The cross-linked products obtained using [14C]cholesterol were characterized by their chromatographic behavior, cleavage on phospholipase A2 treatment, base-catalyzed transesterification and mass spectral measurements. The cross-linking was shown not to involve the 3-beta-hydroxyl group of cholesterol, and it was concluded that the reactive carbene intermediates formed from the photolabels inserted into the hydrocarbon skeleton of cholesterol in the bilayer. The extent of cross-linking obtained was comparable to that observed previously using phospholipids alone, indicating that no lateral phase separation occurred. The present approach is promising for further precise studies of the molecular interactions between cholesterol and phospholipids in biological membranes.


Intercalated cell BKα subunit is required for flow-induced K+ secretion.
BK channels are expressed in intercalated cells (ICs) and principal cells (PCs) in the cortical collecting duct (CCD) of the mammalian kidney and have been proposed to be responsible for flow-induced K+ secretion (FIKS) and K+ adaptation. To examine the IC-specific role of BK channels, we generated a mouse with targeted disruption of the pore-forming BK α subunit (BKα) in ICs (IC-BKα-KO). Whole cell charybdotoxin-sensitive (ChTX-sensitive) K+ currents were readily detected in control ICs but largely absent in ICs of IC-BKα-KO mice. When placed on a high K+ (HK) diet for 13 days, blood [K+] was significantly greater in IC-BKα-KO mice versus controls in males only, although urinary K+ excretion rates following isotonic volume expansion were similar in males and females. FIKS was present in microperfused CCDs isolated from controls but was absent in IC-BKα-KO CCDs of both sexes. Also, flow-stimulated epithelial Na+ channel-mediated (ENaC-mediated) Na+ absorption was greater in CCDs from female IC-BKα-KO mice than in CCDs from males. Our results confirm a critical role of IC BK channels in FIKS. Sex contributes to the capacity for adaptation to a HK diet in IC-BKα-KO mice.


Postural Adjustments and Kinematic Index Finger Features in Frail Older Adults under Different Equilibrium Constraints.
Background: Anticipatory postural adjustments (APAs) are significantly affected by age and may represent restrictions on functional independence. Previous studies in young adults have already highlighted that changing postural stability (i.e., seated vs. upright posture) affects the motor planning and APAs. In frail older adults (FOAs), the effect of these different conditions of postural stability have not yet been established, and the present study aimed to disentangle this issue. Methods: Participants executed an arm-pointing task to reach a diode immediately after it turned on, under different conditions of stability (seated with and without foot support and in an upright posture). A kinematic profile of the index finger and postural electromyographic data were registered in their dominant-side leg muscles: tibialis anterior, soleus, rectus femoris, and semitendinosus. Results: The main finding of this study was that the adopted posture and body stabilization in FOAs did not reflect differences in APAs or kinematic features. In addition, they did not present an optimal APA, since postural muscles are recruited simultaneously with the deltoid. Conclusion: Thus, FOAs seem to use a single non-optimal motor plan to assist with task performance and counterbalance perturbation forces in which they present similar APAs and do not modify their kinematics features under different equilibrium constraints.


Active T-cell receptor genes have intron deoxyribonuclease hypersensitive sites.
The T-cell receptor beta-chain gene has a nuclease hypersensitive site in several kinds of T cells, which does not appear in B cells expressing immunoglobulins. Conversely, the kappa immunoglobulin gene shows a known hypersensitive site at its enhancer element in B cells, as expected, but this site is absent in T cells. As is the case with immunoglobulin genes, the T-cell receptor site lies within the gene, in the intron separating joining and constant region segments. These nuclease hypersensitive DNA configurations in the introns of active T-cell receptor and immunoglobulin genes may arise from control elements that share ancestry but have diverged to the extent that each normally acts only in lymphoid cells which use the proximal gene product.