Exploiting CRISPR-Cas nucleases to produce sequence-specific antimicrobials.
Antibiotics target conserved bacterial cellular pathways or growth functions and therefore cannot selectively kill specific members of a complex microbial population. Here, we develop programmable, sequence-specific antimicrobials using the RNA-guided nuclease Cas9 (refs.1,2) delivered by a bacteriophage. We show that Cas9, reprogrammed to target virulence genes, kills virulent, but not avirulent, Staphylococcus aureus. Reprogramming the nuclease to target antibiotic resistance genes destroys staphylococcal plasmids that harbor antibiotic resistance genes and immunizes avirulent staphylococci to prevent the spread of plasmid-borne resistance genes. We also show that CRISPR-Cas9 antimicrobials function in vivo to kill S. aureus in a mouse skin colonization model. This technology creates opportunities to manipulate complex bacterial populations in a sequence-specific manner.


S100A8 and S100A9: DAMPs at the crossroads between innate immunity, traditional risk factors, and cardiovascular disease.
Amplification of innate immune responses by endogenous danger-associated molecular patterns (DAMPs) promotes inflammation. The involvement of S100A8 and S100A9, DAMPs belonging to the S100 calgranulin family, in the pathogenesis of cardiovascular disease is attracting an increasing amount of interest. S100A8 and S100A9 (also termed MRP8 and MRP14) preferentially form the S100A8/A9 heterodimer (MRP8/14 or calprotectin) and are constitutively expressed in myeloid cells. The levels of circulating S100A8/A9 in humans strongly correlate to blood neutrophil counts and are increased by traditional cardiovascular risk factors such as smoking, obesity, hyperglycemia, and dyslipidemia. S100A8/A9 is an endogenous ligand of toll-like receptor 4 (TLR4) and of the receptor for advanced glycation end products (RAGE) and has been shown to promote atherogenesis in mice. In humans, S100A8/A9 correlates with the extent of coronary and carotid atherosclerosis and with a vulnerable plaque phenotype. S100A8/A9 is locally released following myocardial infarction and amplifies the inflammatory responses associated with myocardial ischemia/reperfusion injury. Elevated plasma levels of S100A8/A9 are associated with increased risk of future coronary events in healthy individuals and in myocardial infarction survivors. Thus, S100A8/A9 might represent a useful biomarker and therapeutic target in cardiovascular disease. Importantly, S100A8/A9 blockers have been developed and are approved for clinical testing.


Slow-light in a vertical-cavity semiconductor optical amplifier.
This paper discusses the effect of slow-light in Vertical-Cavity Semiconductor Optical Amplifiers. A Fabry-Perot model is used to predict the group delay (GD) and GD-bandwidth performance of a VCSOA operated in reflection in the linear regime. It is shown that the GD depends on all cavity parameters while the GDxGD-bandwidth product only depends on the gain. Experimental demonstration with a 1300nm GaInNAs VCSOA is used to validate the model and demonstrate tunable GDs between 25 and 100 ps by varying the VCSOA gain. Experimental distortion of the signals induced by nonlinear effects is also presented.


[Psychological and moral problems in physician-patient relations in carcinoma].
In this article the basic moral and psychological problems a doctor encounters in his professional relationship with a cancer patient are being discussed. That relationship should be mature one, humanistic and sincere. Unfortunately, in the reality it is most frequently an emotionally immature to a certain degree even insincere and thus deprived of its humanistic attributes. The reasons for such a situation are partly concealed in the very psychological problems a doctor might have, which is closely linked to his unconscious motivation for selecting this particular profession as his or her walk of life. The aformentioned problems must be seen in the light of a wide-spread fallacy that the patient need to be preserved from knowing the truth of their doomed state. However, in the same time, there is doctrinary attitude that instructs doctors to reveal all the details describing the situation realistically to the patient but in sincere, thoughtful, warm, humanistic way that can instill some hope to the patient's view on his illness. The mechanisms of phychological defense patients uses when faced to the truth are also being analysed, as well as all therapeutic preventive measures that should have to be taken in addvesing the same issue.


Evolution of herbivory in Drosophilidae linked to loss of behaviors, antennal responses, odorant receptors, and ancestral diet.
Herbivory is a key innovation in insects, yet has only evolved in one-third of living orders. The evolution of herbivory likely involves major behavioral changes mediated by remodeling of canonical chemosensory modules. Herbivorous flies in the genus Scaptomyza (Drosophilidae) are compelling species in which to study the genomic architecture linked to the transition to herbivory because they recently evolved from microbe-feeding ancestors and are closely related to Drosophila melanogaster. We found that Scaptomyza flava, a leaf-mining specialist on plants in the family (Brassicaceae), was not attracted to yeast volatiles in a four-field olfactometer assay, whereas D. melanogaster was strongly attracted to these volatiles. Yeast-associated volatiles, especially short-chain aliphatic esters, elicited strong antennal responses in D. melanogaster, but weak antennal responses in electroantennographic recordings from S. flava. We sequenced the genome of S. flava and characterized this species' odorant receptor repertoire. Orthologs of odorant receptors, which detect yeast volatiles in D. melanogaster and mediate critical host-choice behavior, were deleted or pseudogenized in the genome of S. flava. These genes were lost step-wise during the evolution of Scaptomyza. Additionally, Scaptomyza has experienced gene duplication and likely positive selection in paralogs of Or67b in D. melanogaster. Olfactory sensory neurons expressing Or67b are sensitive to green-leaf volatiles. Major trophic shifts in insects are associated with chemoreceptor gene loss as recently evolved ecologies shape sensory repertoires.


Photostimulation of Phox2b medullary neurons activates cardiorespiratory function in conscious rats.
Hypoventilation is typically treated with positive pressure ventilation or, in extreme cases, by phrenic nerve stimulation. This preclinical study explores whether direct stimulation of central chemoreceptors could be used as an alternative method to stimulate breathing. To determine whether activation of the retrotrapezoid nucleus (RTN), which is located in the rostral ventrolateral medulla (RVLM), stimulates breathing with appropriate selectivity. A lentivirus was used to induce expression of the photoactivatable cationic channel channelrhodopsin-2 (ChR2) by RVLM Phox2b-containing neurons, a population that consists of central chemoreceptors (the ccRTN neurons) and blood pressure (BP)-regulating neurons (the C1 cells). The transfected neurons were activated with pulses of laser light. Respiratory effects were measured by plethysmography or diaphragmatic EMG recording and cardiovascular effects by monitoring BP, renal sympathetic nerve discharge, and the baroreflex. The RVLM contained 600 to 900 ChR2-transfected neurons (63% C1, 37% ccRTN). RVLM photostimulation significantly increased breathing rate (+42%), tidal volume (21%), minute volume (68%), and peak expiratory flow (48%). Photostimulation increased diaphragm EMG amplitude (19%) and frequency (21%). Photostimulation increased BP (4 mmHg) and renal sympathetic nerve discharge (43%) while decreasing heart rate (15 bpm). Photostimulation of ChR2-transfected RVLM Phox2b neurons produces a vigorous stimulation of breathing accompanied by a small sympathetically mediated increase in BP. These results demonstrate that breathing can be relatively selectively activated in resting unanesthetized mammals via optogenetic manipulation of RVLM neurons presumed to be central chemoreceptors. This methodology could perhaps be used in the future to enhance respiration in humans.


Single stranded DNA SP6 promoter plasmids for engineering mutant RNAs and proteins: synthesis of a 'stretched' preproparathyroid hormone.
The intergenic region of bacteriophage f1 has been subcloned into the bacteriophage SP6 promoter plasmids, pSP64 and pSP65, in both orientations. Coinfection of E. coli with these SP6 promoter/phage f1 chimeric plasmids and the interference resistance phage, IR1, results in the replication and secretion of the pSP6.f1 plasmids as single stranded DNA. Bovine preProPTH cDNAs in both the native form and a form containing an insertion of 117 base pairs in the protein coding region have been inserted in these plasmids. The RNA transcribed from the SP6.f1/preProPTH cDNA constructs was efficiently translated in the wheat germ or reticulocyte cell free systems without addition of a 7-methylguanosine cap to the RNA. In the presence of dog pancreatic or chicken oviduct microsomal membranes, conversion of the resultant pre-proteins to pro-proteins was observed. Confirmation of the "mutated" preProPTH cDNA was determined by dideoxyribonucleotide DNA sequencing of single stranded plasmid DNA. These vectors are suitable for the efficient biosynthesis of large amounts of single or double stranded DNA, and translationally active RNA. The combined properties of single stranded DNA replication and the SP6 promoter simplify the engineering of mutant RNAs and their corresponding proteins. In addition, single stranded DNA or RNA corresponding to either complementary strand may be synthesized as nucleic acid hybridization probes.


Skin temperature changes induced by strong static magnetic field exposure.
High intensity static magnetic fields, when applied to the whole body of the anesthetized rat, have previously been reported to decrease skin temperature. The hypothesis of the present study was that in diamagnetic water, molecules in the air play significant roles in the mechanism of skin temperature decrease. We used a horizontal cylindrical superconducting magnet. The magnet produced 8 T at its center. A thermistor probe was inserted in a subcutaneous pocket of the anesthetized rats to measure skin temperature. Animals (n=10) were placed in an open plastic holder in which the ambient air was free to move in any direction (group I). Animals (n=10) were placed in a closed holder in which the air circulation toward the direction of weak magnetic field was restricted (group II). Each holder was connected to a hydrometer to measure humidity around the animal in the holder. The data acquisition phase consisted of a 5 min baseline interval, followed by inserting the animal together with the holder into the center of the magnet bore for a 5 min exposure and a 5 min postexposure period outside the bore. In group I, skin temperature and humidity around the animal significantly decreased during exposure, followed by recovery after exposure. In group II, skin temperature and humidity did not decrease during the measurement. The skin temperature decrease was closely related to the decrease in humidity around the body of the animal in the holder, and the changes were completely blocked by restricting the air circulation in the direction of the bore entrance. Possible mechanisms responsible for the decrease in skin temperature may be associated with magnetically induced movement of water vapor at the skin surface, leading to skin temperature decrease.


The prognosis of migraine.
The prognosis of migraine is poorly studied and research in this regard is in its infancy. Migraine is a chronic disorder with episodic attacks with a highly variable long-term prognosis. In many, migraine may have a very benign (complete remission) or relatively benign (partial remission) prognosis. In some, migraine persists and in others, it progresses. A recent population study showed that, over a 1-year period, 84% of the patients with migraine persisted with this diagnosis (migraine persistence); around 10% had 1-year complete clinical remission, and 3% had partial remission; the other 3% developed chronic migraine. Long-term studies support the concept that remission increases with age and also that risk factors for progression have been identified (e.g. medication overuse, obesity, etc.). Identifying predictors of migraine prognosis is, therefore, of great scientific and clinical interest. Herein we review the prognosis of migraine and risk factors for the above-mentioned nonexclusive possibilities.


Human leukocyte antigen-E protein is overexpressed in primary human colorectal cancer.
HLA-E is a non-classical MHC molecule whose expression by tumour cells has been recently reported in several human cancer types. We studied HLA-E expression in colorectal cancer patients, its clinical significance and prognostic value, as well as characterized its expression in colorectal cancer cell lines. We analysed HLA-E expression at the transcript level by qRT-PCR in micro-dissected samples and at the protein level by semiquantitative immunohistochemistry on paraffin-embedded tissue sections from 42 biopsies of colorectal cancer patients. We observed that HLA-E transcript and protein are spontaneously overexpressed in a significant proportion of colorectal tumour biopsies, as compared to normal mucosae. We also found a negative correlation between HLA-E expression and the CD57+ cells infiltrate. Moreover, we analysed HLA-E expression in several colorectal cancer cell lines and demonstrated that IFN-gamma upregulates the expression of membrane HLA-E in vitro. Interestingly, we demonstrated that colorectal cancer cell lines overexpressing HLA-E at the cell surface inhibited NK-mediated cell lysis. Although IFN-gamma regulatory role needs further investigation, we provide evidence suggesting that this cytokine, within the tumour microenvironment, could promote HLA-E translocation to the surface of tumour epithelial cells. Furthermore, we showed that upregulation of HLA-E could be a marker of shorter disease-free survival in Dukes' C patients and we suggest that this molecule renders tumours less susceptible to immune attack.


Impact of Comorbidities and Commonly Used Drugs on Mortality in COPD - Real-World Data from a Primary Care Setting.
Life expectancy is significantly shorter for patients with chronic obstructive pulmonary disease (COPD) than the general population. Concurrent diseases are known to infer an increased mortality risk in those with COPD, but the effects of pharmacological treatments on survival are less established. This study aimed to examine any associations between commonly used drugs, comorbidities and mortality in Swedish real-world primary care COPD patients. Patients with physician-diagnosed COPD from a large primary care population were observed retrospectively, utilizing primary care records and mandatory Swedish national registers. The time to all-cause death was assessed in a stepwise multiple Cox proportional hazards regression model including demography, socioeconomic factors, exacerbations, comorbidities and medication. During the observation period (1999-2009) 5776 (32.5%) of 17,745 included COPD patients died. Heart failure (hazard ratio [HR]: 1.88, 95% confidence interval [CI]: 1.74-2.04), stroke (HR: 1.52, 95% CI: 1.40-1.64) and myocardial infarction (HR: 1.40, 95% CI: 1.24-1.58) were associated with an increased risk of death. Use of inhaled corticosteroids (ICS; HR: 0.79, 95% CI: 0.66-0.94), beta-blockers (HR: 0.86, 95% CI: 0.76-0.97) and acetylsalicylic acid (ASA; HR: 0.87, 95% CI: 0.77-0.98) was dose-dependently associated with a decreased risk of death, whereas use of long-acting muscarinic antagonists (LAMA; HR: 1.33, 95% CI: 1.14-1.55) and N-acetylcysteine (NAC; HR: 1.26, 95% CI: 1.08-1.48) were dose-dependently associated with an increased risk of death in COPD patients. This large, retrospective, observational study of Swedish real-world primary care COPD patients indicates that coexisting heart failure, stroke and myocardial infarction were the strongest predictors of death, underscoring the importance of timely recognition and treatment of comorbidities. A decreased risk of death associated with the use of ICS, beta-blockers and ASA, and an increased risk associated with the use of LAMA and NAC, was also found.


Immunochemical characterization and purification of Sm-97, a Schistosoma mansoni antigen monospecifically recognized by antibodies from mice protectively immunized with a nonliving vaccine.
Mice protected against Schistosoma mansoni infection by intradermal (i.d.) vaccination with nonliving schistosomula or soluble extracts of larval or adult schistosomes (SCHLAP and SWAP, respectively) produce antibodies that react by Western blot analysis with one antigen of Mr (X 10(-3)) 97 in SWAP prepared in the presence of protease inhibitors and two antigens of Mr (X 10(-3)) 95 and 78 in SWAP prepared in their absence. Vaccine antibodies also immunoprecipitated a single 97k molecule, with a pI of 5.5, from detergent extracts of [35S] methionine-labeled schistosomes. Three hybridomas, produced from spleen cells of i.d. immunized mice, all recognized both the 95k/78k doublet and the 97k antigen, indicating that the two lower Mr components are degradation products of the same 97k molecule. The 97k/95k/78k complex (Sm-97) was purified by affinity chromatography and found to constitute 0.5% of the total protein in SWAP. 125I-concanavalin A bound weakly to purified Sm-97, indicating that this antigen is minimally glycosylated. By indirect immunofluorescence, Sm-97 was localized to regions just below the tegumental and gut syncitia of adult worms. Mice protected by i.d. vaccination produced high titers (1:10,240) of anti-Sm-97 antibodies, whereas chronically infected mice responded at a much lower level (titer 1:640). In contrast, mice protectively immunized with irradiated cercariae and mice nonprophylactically inoculated by the i.v. route failed to produce detectable anti-Sm-97 antibodies. Competitive radioimmunoassays performed with 125I-labeled monoclonal antibodies and purified antigen defined at least two distinct epitopes on Sm-97. Antibodies from i.d. vaccinated mice recognized both monoclonal antibody-defined epitopes, whereas anti-Sm-97 antibodies in chronic infection sera recognized neither. Finally, purified Sm-97 was shown to elicit delayed-type hypersensitivity in i.d. vaccinated mice, suggesting that this molecule is also capable of evoking cell-mediated responses, a finding consistent with its proposed function as a vaccine immunogen.


Anti-enterovirus 71 activities of Melissa officinalis extract and its biologically active constituent rosmarinic acid.
Enterovirus 71 (EV71) infection is endemic in the Asia-Pacific region. No specific antiviral drug has been available to treat EV71 infection. Melissa officinalis (MO) is a medicinal plant with long history of usage in the European and Middle East. We investigated whether an aqueous solution of concentrated methanolic extract (MOM) possesses antiviral activity. MOM inhibited plaque formation, cytopathic effect, and viral protein synthesis in EV71-infected cells. Using spectral techniques, we identified rosmarinic acid (RA) as a biologically active constituent of MOM. RA reduced viral attachment and entry; cleavage of eukaryotic translation initiation factor 4 G (eIF4G); reactive oxygen species (ROS) generation; and translocation of heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1) from nucleus to cytoplasm. It alleviated EV71-induced hyperphosphorylation of p38 kinase and EPS15. RA is likely to suppress ROS-mediated p38 kinase activation, and such downstream molecular events as hnRNP A1 translocation and EPS15-regulated membrane trafficking in EV71-infected cells. These findings suggest that MO and its constituent RA possess anti-EV71 activities, and may serve as a candidate drug for therapeutic and prophylactic uses against EV71 infection.


A water molecule participates in the secondary structure of hyaluronan.
The structure of hyaluronan was investigated in water/dimethyl sulphoxide mixtures by using high-field n.m.r. and space-filling molecular models. The secondary structure previously established in detail in 'dry' dimethyl sulphoxide [Heatley, Scott & Hull (1984) Biochem. J. 220, 197-205] undergoes changes on addition of water, compatible with the incorporation of a water bridge between the uronate carboxylate and acetamido NH groups. Molecular models show that such a configuration is highly probable, and saturation-transfer experiments yield rates of NH proton exchange that support this proposed structure. The existence of two distinct stable configurations for hyaluronan, in water-rich and water-poor conditions respectively, may have biological implications, e.g. during its biosynthesis in cell membranes. There are extensive hydrophobic regions in both forms, which may be important for interactions with e.g., membranes, proteins and itself.


Palindrome resolution and recombination in the mammalian germ line.
Genetic instability is promoted by unusual sequence arrangements and DNA structures. Hairpin DNA structures can form from palindromes and from triplet repeats, and they are also intermediates in V(D)J recombination. We have measured the genetic stability of a large palindrome which has the potential to form a one-stranded hairpin or a two-stranded cruciform structure and have analyzed recombinants at the molecular level. A palindrome of 15.3 kb introduced as a transgene was found to be transmitted at a normal Mendelian ratio in mice, in striking contrast to the profound instability of large palindromes in prokaryotic systems. In a significant number of progeny mice, however, the palindromic transgene is rearranged; between 15 and 56% of progeny contain rearrangements. Rearrangements within the palindromic repeat occur both by illegitimate and homologous, reciprocal recombination. Gene conversion within the transgene locus, as quantitated by a novel sperm fluorescence assay, is also elevated. Illegitimate events often take the form of an asymmetric deletion that eliminates the central symmetry of the palindrome. Such asymmetric transgene deletions, including those that maintain one complete half of the palindromic repeat, are stabilized so that they cannot undergo further illegitimate rearrangements, and they also exhibit reduced levels of gene conversion. By contrast, transgene rearrangements that maintain the central symmetry continue to be unstable. Based on the observed events, we propose that one mechanism promoting the instability of the palindrome may involve breaks generated at the hairpin structure by a hairpin-nicking activity, as previously detected in somatic cells. Because mammalian cells are capable of efficiently repairing chromosome breaks through nonhomologous processes, the resealing of such breaks introduces a stabilizing asymmetry at the center of the palindrome. We propose that the ability of mammalian cells to eliminate the perfect symmetry in a palindromic sequence may be an important DNA repair pathway, with implications regarding the metabolism of palindromic repeats, the mutability of quasipalindromic triplet repeats, and the early steps in gene amplification events.


Provider-initiated HIV testing and counseling in TB clinical settings: tools for program implementation.
In countries with high HIV prevalence, up to 80% of tuberculosis (TB) patients also have human immunodeficiency virus (HIV) infection. HIV testing and counseling needs to be more feasible and accessible to all people in settings with a generalized HIV epidemic, but particularly to those likely to have HIV infection and who need care and treatment. Implementing provider-initiated and -delivered HIV testing and counseling (PITC) in clinical settings where patients have symptoms and signs consistent with HIV-related disease, including TB, is therefore a priority. We describe a new tool that has been developed to assist countries in planning and implementing PITC in TB clinical settings. The materials include a template for national guidance for the PITC program and procedures, a training curriculum for clinic staff and job aids including a script that assist clinicians in communicating appropriate pre- and post-test information to their TB patients, including the benefits to HIV-infected TB patients of knowing their status so they can obtain HIV care and treatment and prevent the spread of HIV.


The Prognostic Value of Immune Factors in the Tumor Microenvironment of Penile Squamous Cell Carcinoma.
The host's immune system plays a pivotal role in many tumor types, including squamous cell carcinomas (SCCs). We aim to identify immunological prognosticators for lymph node metastases (LNM) and disease-specific survival (DSS) in penile SCC. For this retrospective observational cohort study, penile SCC patients (n = 213) treated in the Netherlands Cancer Institute, were selected if sufficient formalin-fixed, paraffin-embedded tumor material was available. Analysis included previously described high-risk human papilloma virus (hrHPV) status, immunohistochemical scores for classical and non-classical human leukocyte antigen (HLA) class I, programmed death ligand-1 (PD-L1) expression, and novel data on tumor-infiltrating macrophages and cytotoxic an regulatory T-cells. Clinicopathological characteristics and extended follow-up were also included. Regression analyses investigated relationships of the immune parameters with LNM and DSS. In the total cohort, diffuse PD-L1 tumor-cell expression, CD163+ macrophage infiltration, non-classical HLA class I upregulation, and low stromal CD8+ T-cell infiltration were all associated with LNM. In the multivariable model, only tumor PD-L1 expression remained a significant predictor for LNM (odds ratio (OR) 2.8, p = 0.05). hrHPV negativity and diffuse PD-L1 tumor-cell expression were significantly associated with poor DSS and remained so upon correction for clinical parameters [hazard ratio (HR) 9.7, p < 0.01 and HR 2.8, p = 0.03]. The only immune factor with different expression in HPV+ and HPV- tumors was PD-L1, with higher PD-L1 expression in the latter (p = 0.03). In the HPV- cohort (n = 158), LNM were associated with diffuse PD-L1 tumor-cell expression, high intratumoral CD163+ macrophage infiltration, and low number of stromal CD8+ T-cells. The first two parameters were also linked to DSS. In the multivariable regression model, diffuse PD-L1 expression remained significantly unfavorable for DSS (HR 5.0, p < 0.01). These results emphasize the complexity of the tumor microenvironment in penile cancer and point toward several possible immunotherapy targets. Here described immune factors can aid risk-stratification and should be evaluated in clinical immunotherapy studies to ultimately lead to patient tailored treatment.


Advances in the elegance of chemistry in designing dendrimers.
Highly functional and monodisperse macromolecules with tailored architecture constitute the key to designing efficient and smart nanomaterials. Dendrimers offer real potential to achieve this goal, and one of the earlier challenges faced by this novel class of polymers has been addressed by the evolution of synthetic methodologies. This review provides an evaluation of the role played by chemistry in taking these macromolecules of academic relevance to practical industrial and biological applications, in a relatively short period. One can now construct dendrimers in a 'made-to-order' fashion, for numerous applications in a variety of disciplines.


Pharmacokinetic Modeling and Limited Sampling Strategies Based on Healthy Volunteers for Monitoring of Ertapenem in Patients with Multidrug-Resistant Tuberculosis.
Ertapenem is a broad-spectrum carbapenem antibiotic whose activity against Mycobacterium tuberculosis is being explored. Carbapenems have antibacterial activity when the plasma concentration exceeds the MIC at least 40% of the time (40% TMIC). To assess the 40% TMIC in multidrug-resistant tuberculosis (MDR-TB) patients, a limited sampling strategy was developed using a population pharmacokinetic model based on data for healthy volunteers. A two-compartment population pharmacokinetic model was developed with data for 42 healthy volunteers using an iterative two-stage Bayesian method. External validation was performed by Bayesian fitting of the model developed with data for volunteers to the data for individual MDR-TB patients (in which the fitted values of the area under the concentration-time curve from 0 to 24 h [AUC0-24, fit values] were used) using the population model developed for volunteers as a prior. A Monte Carlo simulation (n = 1,000) was used to evaluate limited sampling strategies. Additionally, the 40% TMIC with the free fraction (f 40% TMIC) of ertapenem in MDR-TB patients was estimated with the population pharmacokinetic model. The population pharmacokinetic model that was developed was shown to overestimate the area under the concentration-time curve from 0 to 24 h (AUC0-24) in MDR-TB patients by 6.8% (range, -17.2 to 30.7%). The best-performing limited sampling strategy, which had a time restriction of 0 to 6 h, was found to be sampling at 1 and 5 h (r2 = 0.78, mean prediction error = -0.33%, root mean square error = 5.5%). Drug exposure was overestimated by a mean percentage of 4.2% (range, -15.2 to 23.6%). When a free fraction of 5% was considered and the MIC was set at 0.5 mg/liter, the minimum f 40% TMIC would have been exceeded in 9 out of 12 patients. A population pharmacokinetic model and limited sampling strategy, developed using data from healthy volunteers, were shown to be adequate to predict ertapenem exposure in MDR-TB patients.


Use of psychoactive substances by medical students: a survey.
The use and abuse of alcohol and other psychoactive substances is of concern to the medical profession and to medical educators. Sixty-eight members of the sophomore class at LSU Medical Center in Shreveport participated in a substance use survey. Ten students (14.7%) described themselves as abstainers from alcohol consumption. Although none described themselves as heavy or abusive drinkers, 9 (13.2%) drank 6-10 alcoholic beverages on a weekend if there was no examination scheduled for Monday or Tuesday, and 3 (4.4%) consumed more than 10 alcoholic beverages on such a weekend. At a large school social event, 9 students (13.2%) drank 6-10 alcoholic beverages and 3 (4.4%) drank more than 10 alcoholic beverages during the event. Nine students (13.2%) used marijuana at least 1-3 times per year. Tobacco was used by 17.6% of the students sampled. Only 4 students (5.9%) avoided caffeine completely. Twelve students (17.6%) drank 5-10 caffeinated beverages daily. The use of cocaine (2.9%), amphetamines (4.5%), and unprescribed sedatives (6%) was small in this student sample. More education concerning effects of psychoactive substances is encouraged.


A van der Waals density functional investigation of carboranethiol self-assembled monolayers on Au(111).
Isolated and full monolayer adsorption of various carboranethiol (C2B10H12S) isomers on the gold(111) surface has been investigated using both the standard and van der Waals density functional theory calculations. The effect of different molecular dipole moment orientations on the low energy adlayer geometries, the binding characteristics and the electronic properties of the self-assembled monolayers of these isomers has been studied. Specifically, the binding energy and work function changes associated with different molecules show a correlation with their dipole moments. The adsorption is favored for the isomers with dipole moments parallel to the surface. Of the two possible unit cell structures, (5 × 5) was found to be more stable than .


Perspectives for Remote Sensing with Unmanned Aerial Vehicles in Precision Agriculture.
Remote sensing with unmanned aerial vehicles (UAVs) is a game-changer in precision agriculture. It offers unprecedented spectral, spatial, and temporal resolution, but can also provide detailed vegetation height data and multiangular observations. In this article, we review the progress of remote sensing with UAVs in drought stress, in weed and pathogen detection, in nutrient status and growth vigor assessment, and in yield prediction. To transfer this knowledge to everyday practice of precision agriculture, future research should focus on exploiting the complementarity of hyperspectral or multispectral data with thermal data, on integrating observations into robust transfer or growth models rather than linear regression models, and on combining UAV products with other spatially explicit information.


Comparing hands-on and video training for postpartum hemorrhage management.
The objective was to compare two teaching methods for postpartum hemorrhage management: interactive hands-on training and non-interactive video training. In a controlled intervention study at a secondary health care center in Kenya, the two training methods, based on the Advanced Life Support in Obstetrics curriculum, were evaluated utilizing structured observation of a standardized scenario before and after training. Both intervention groups significantly increased in performance scores after receiving hands-on training: 40% (95% CI 29.5-47.0) and video training: 34.5% (95% CI 25.0-42.0); likewise, pass rates improved significantly. No significant differences in performance score or pass rates were found between the two methods. The findings indicate that postpartum hemorrhage management training by mobile media might be just as effective as conventional hands-on training and a feasible way to overcome the outreach gap in sub-Saharan Africa's rural areas, where peripheral health facilities are generally difficult to reach with conventional training programs.


Pressure-Induced Glass Transition Probed via the Mobility of Coumarin 1 Fluorescent Molecule.
The route to form a glass is generally achieved upon cooling where the slowing down might be interpreted as the trapping of molecules in potential wells. On the other hand, isothermal compression induces a glassy state by modifying the molecular packing ending in jamming. Here, we focus on how isothermal compression perturbs the mobility of a probe molecule in three different host liquids up to the pressure-induced glass transition. By use of the fluorescence recovery technique, the diffusion of the fluorescent molecule Coumarin 1 (C1) is measured in poly(propylene glycol) (PPG-1000M and -2700M), in the fragile van der Waals propylene carbonate (PC), and in hydrogen-bonded methanol and ethanol. High pressures up to 6 GPa are obtained with a diamond anvil cell. In PC at a pressure ∼1.3 GPa close to the glass-transition pressure, the diffusion coefficient of C1 follows an Arrhenius behavior with an ∼5 orders of magnitude increase of the diffusive time. No decoupling from the Stokes-Einstein equation is noticed. A similar exponential behavior is measured in ethanol and methanol but extended to different pressure ranges up to 2.5 and 6.2 GPa, respectively. In PPG-1000M a decoupling from the Stokes-Einstein relation is observed between 0.3 and 0.8 GPa that could be related to a modification of the interaction between polymer segments and the probe molecule. These results might indicate that interaction between probe and dynamic heterogeneities become less important under applied pressure, unlike in the temperature-induced glass transition.


Irritable Bowel Syndrome in Iran: SEPAHAN Systematic Review No. 1.
Irritable bowel syndrome (IBS) is a common gastrointestinal disorder. Due to its high prevalence and absence of curative therapy, IBS has the potential to create tremendous burden on the health care system. Herein, we systematically reviewed the published literature to investigate the epidemiology of IBS in Iran. Studies that were reviewed in this article were primarily identified through four online bibliographic databases including PubMed, Google Scholar, Iran Medex, and Scientific Information Database. Manual search of reference lists was carried out to identify any additional studies such as relevant abstracts and also recent review articles which may have been missed. Potentially related studies were retrieved and the selection criteria were applied. Eligible articles were reviewed. From 4176 studies identified, 18 eligible studies were included. It was reported that in Iran, the prevalence of IBS was in the range of 1.1% to 25% and was more common in women. In addition, the difference in frequency of different age groups was minimal. There was a minimal difference in IBS prevalence within different age groups. In Iran, the incidence of IBS was in the wide range. Since there are not enough population-based studies, researchers should focus on developing well-designed population-based studies to determine the epidemiology of IBS in Iran. Moreover, cohort studies should be conducted in order to investigate the natural history of IBS. Investigating the etiology of IBS and attempt to organize health promotion programs are highly suggested.


Apoptotic Effects of Melittin on 4T1 Breast Cancer Cell Line is associated with Up Regulation of Mfn1 and Drp1 mRNA Expression.
Melittin, as the main ingredient of honeybee venom, that has shown anticancer properties. The present study aimed at investigating the cytotoxic impacts of melittin on 4T1 breast cancer cells. Hemolytic activity of different concentrations (0.125, 0.25, 0.5, 1, 2, 4, 8μg/ml) of melittin was assayed and then cytotoxicity of selected concentrations of melittin (2, 4, 8, 16, 32, and 64μg/ml), 2 and 4μg/ml of cisplatin and 0.513, 0.295 and 0.123μg/ml of doxorubicin was evaluated on 4T1 cells using MTT assay. We used Morphological evaluation and flow cytometric analysis was used. Real time PCR was also used to determine mRNA expression of Mfn1 and Drp1 genes. All compounds showed anti-proliferative effects on the tumor cell line with different potencies. Melittin had higher cytotoxicity against 4T1 breast cancer cells (IC50= 32μg/ml-72h) and higher hemolytic activity (HD50= 1μg/ml), as compared to cisplatin and doxorubicin. Mellitin at 16 and 32μg/ml showed apoptotic effects on 4T1 cells according to the flow cytometric analysis. The Real time PCR analysis of Drp1 and Mfn1 expression in cells treated with 16μg/ml of melittin revealed an up-regulation in Drp1 and Mfn1 genes mRNA expression in comparison with control group. Treatment with 32μg/ml of melittin was also associated with a rise in mRNA expression of Drp1 and Mfn1 as compared to the control group. The results of this study showed that melittin has anticancer effects on 4T1 cell lines in a dose and time dependent manner and can be a good candidate for further research on breast cancer treatment.


New era of liver transplantation for hepatitis B: a 17-year single-center experience.
To evaluate the variables affecting orthotopic liver transplantation (OLT) outcome for hepatitis B virus (HBV) in a large patient cohort over a 17-year period. Historically, OLT for chronic HBV infection has been associated with aggressive reinfection and poor survival results. More recently, OLT outcome has been improved with the routine use of antiviral therapy with either hepatitis B immune globulin (HBIg) or lamivudine; however, HBV recurrence remains common. The authors studied the factors affecting HBV recurrence and outcome of transplantation, including the effects of combination viral prophylaxis with HBIg and lamivudine. A retrospective review of 166 OLT recipients for chronic HBV over a 17-year period at a single center was performed. Median follow-up was 29 months. HBV recurrence was defined by HBsAg seropositivity after OLT. HBIg monotherapy was used in 28 (17%) patients, lamivudine monotherapy in 20 (12%), and HBIg and lamivudine combination in 89 (54%); 29 (17%) did not receive any HBV prophylaxis. Hepatocellular carcinoma (HCC) was present in 43 patients (26%) and urgent United Network for Organ Sharing (UNOS) status was assigned to 27 patients (16%). Univariate and multivariate analyses were performed to identify factors that affected OLT outcome. Overall 1-, 3-, and 5-year patient survival rates were 85.8%, 73.6%, and 71.8%, respectively. As expected, HBV recurrence-free survival rates were significantly lower than overall survival rates (76.4%, 58.7%, and 48.3%). When compared with a nontreated cohort, OLT recipients receiving combination viral prophylaxis with HBIg and lamivudine showed markedly reduced HBV recurrence rates and significantly improved 1- and 3-year recurrence-free survival rates. By univariate estimates, patient survival was reduced in the presence of HCC, in the Asian population, and urgent candidates by UNOS classification. Graft loss rates were significantly increased in urgent OLT candidates, Asians, patients with pretransplant positive DNA, and in the presence of HCC. Factors that were significant by univariate analysis or thought to be clinically relevant were subjected to multivariate analysis. By multivariate estimates, urgent UNOS or presence of HCC adversely affected patient and graft survival rates, whereas combination prophylactic therapy strongly predicted improved patient and graft survival rates as well as recurrence-free survival rates. Orthotopic liver transplantation for HBV under combination viral prophylaxis results in survival rates equivalent to other indications. Pretransplant viral replication, UNOS status, and the presence of HCC are all sensitive markers for posttransplantation outcome. Viral prophylactic therapy has effectively reduced HBV recurrence and prolonged survival outcomes. The combination of HBIg and lamivudine is the prophylactic regimen of choice.


Loeys-Dietz syndrome in pregnancy.
Loeys-Dietz syndrome is a recently described condition which causes cardiovascular, craniofacial, neurocognitive and skeletal abnormalities due to mutations in components of the transforming growth factor-β signalling pathway. Associated vascular abnormalities include vessel tortuosity and an increased incidence of vascular dissection. Pregnancy increases the risk of aortic dissection compared to non-pregnant individuals and an underlying condition such as Loeys-Dietz syndrome increases this further. While aortic dissection is well described in pregnancy in Loeys-Dietz syndrome, some women can have uncomplicated deliveries, particularly when the risks of the condition are actively managed. Such pregnancies should be considered high-risk, and women should be counselled and managed accordingly. Here we describe two pregnancies in one woman, both with successful outcomes, followed by a summary of the key management principles.


On the nature of blueshifting hydrogen bonds.
The block-localized wave function (BLW) method can derive the energetic, geometrical, and spectral changes with the deactivation of electron delocalization, and thus provide a unique way to elucidate the origin of improper, blueshifting hydrogen bonds versus proper, redshifting hydrogen bonds. A detailed analysis of the interactions of F(3)CH with NH(3) and OH(2) shows that blueshifting is a long-range phenomenon. Since among the various energy components contributing to hydrogen bonds, only the electrostatic interaction has long-range characteristics, we conclude that the contraction and blueshifting of a hydrogen bond is largely caused by electrostatic interactions. On the other hand, lengthening and redshifting is primarily due to the short-range n(Y)→σ*(X-H) hyperconjugation. The competition between these two opposing factors determines the final frequency change direction, for example, redshifting in F(3)CH⋅⋅⋅NH(3) and blueshifting in F(3)CH⋅⋅⋅OH(2). This mechanism works well in the series F(n)Cl(3)-n CH⋅⋅⋅Y (n=0-3, Y=NH(3), OH(2), SH(2)) and other systems. One exception is the complex of water and benzene. We observe the lengthening and redshifting of the O-H bond of water even with the electron transfer between benzene and water completely quenched. A distance-dependent analysis for this system reveals that the long-range electrostatic interaction is again responsible for the initial lengthening and redshifting.


[CONSEQUENCES OF FALLS IN A GERIATRIC NURSING CENTER FOR VALID. DESCRIPTIVE STUDY].
To address the immediate consequences produced in the falls suffered by valid institutionalized elderly population' in the last twelve months. A cross sectional study, by personal interview. Simple random sampling was performed. The study area corresponded to a private nursing home. The measuring instruments used in the study were the Mini Mental State Examination Lobo (MEC-35, 1979) to initially assess the level of cognitive development elderly study participants, and the WHO questionnaire for the study of falls in the elder 1989. Of the 21 drops produced in the last twelve months was found that 61.9% of falls were female, compared to 38.1% who relapsed in males. 52.4% of falls are not caused any consequences. 28.6% of falls caused superficial injuries and/or bruises, while the proportion of fractures corresponded to 19% for both sexes. Most of the falls did not produce immediate consequences whose percentage was 52.4% of cases. Second are superficial wounds and/or bruises with 28.6% and finally fractures with a ratio of 19%. The investigators considered the necessity of involvement of health professionals and the implementation ofpreventive strategies that can reduce the prevalence of falls. Thiswill successfully improve the quality of life of the elderly.


Linear dimension reduction of sequences of medical images: III. Factor analysis in signal space.
A method is presented for improving the precision of factor analysis by utilizing physiological information. The first step is an optimal linear dimension reduction, whereby the data are projected onto a low-dimensional signal space. Then principal component analysis is performed in the signal space rather than in the entire data space. This improves the precision of the principal components. Unlike ordinary principal component analysis, the present method is not degraded when the time intervals are subdivided, provided that the signal space is correct. Alternatively, but with identical results, the covariance matrix can be calculated from the whole data space. The covariance matrix is then transformed and principal component analysis is performed in either a low-rank matrix or a low-dimensional submatrix instead of in the whole covariance matrix. Factor analysis using the intersection method with a theory space may be improved by employing the present method. In simulations based on a [11C]flumazenil study with 27 frames, the proposed method required only 58 per cent of the radioactivity to produce the same precision as the intersection method and only 27 per cent when compared to ordinary principal component analysis.


Sexual behavior decreases pain sensitivity and stimulated endogenous opioids in male rats.
In male rats copulation has antinociceptive effects as measured either by shock-induced vocalizations or hindlimb withdrawal to pinch. Prolonged mating reduces the content of endogenous opioids in midbrain but not in hypothalamus or caudate nucleus. Blockage of opiate receptors with the narcotic antagonist naloxone (4 mg/kg) significantly extends the postejaculatory interval. The results indicate that mating is a biological stimulus for the release of endogenous opoids, possibly to (a) prevent intense sexual stimulation from becoming aversive, and (b) increase its reward value.


Acoustic analysis of speech intonation pattern of individuals with Autism Spectrum Disorders.
This study aimed to analyze prosodic elements of speech segments of students with Autism Spectrum Disorders (ASD) and compare with the control group, using an acoustic analysis. Speech recordings were performed with a sample of individuals with ASD (n = 19) and with typical development (n = 19) of the male gender, age range: 8-33 years. The prosody questionnaire ALIB (Brazilian Linguistic Atlas) was used as script, which contains interrogative, affirmative and imperative sentences. Data were analyzed using the PRAAT software and forwarded to statistical analysis in order to verify possible significant differences between the two groups studied in each prosodic parameter evaluated (fundamental frequency, intensity and duration) and its respective variables. There were significant differences for the variables tessitura, melodic amplitude of tonic vowel, melodic amplitude of pretonic vowel, maximum intensity, minimum intensity, tonic vowel duration, pretonic vowel duration and phrase duration. Individuals with ASD present significant differences in prosody compared to those with typical development. It is noteworthy, however, the necessity of additional studies on the characterization of prosodic aspects of speech of individuals with ASD with a larger sample and a more restricted age group.


The highly elongated Drosophila mechanosensory bristle--a new model for studying polarized microtubule function.
Microtubules (MTs) are polar polymers that can facilitate asymmetric distribution of cell components, a process important for polarized cell growth. The highly elongated and polarized Drosophila mechnosensory bristle cytoplasm is filled with short MTs that constitute a significant component of the shaft cytoplasm. Inhibition of MT assembly affects biased axial growth in the bristle and highlights the importance of MTs for this process. We demonstrate that the vast majority of bristle MTs are organized in a polarized manner, minus-ends out. We also show that genetic disruption of the MT polarity affects the polar distribution of cell components and leads to an alteration in the biased axial shape of the bristle shaft. Thus, we suggest that the asymmetric organization of the MT population within the bristle cell shaft is necessary for the proper axial elongation of this cellular extension. We would also like to emphasize the benefits of using the bristle cell as a model for studying MTs and MT-binding proteins because changes to this cytoskeletal component result in easily recognized at the phenotypes.


Methods for invasive species control are transferable across invaded areas.
Cane Toads (Rhinella marina) are invasive pests in many parts of the world, including the Japanese island of Ishigaki. Extensive research in Australia has identified promising new methods for control, but also has shown that toads exhibit geographic variation in many traits (suggesting that methods developed in one location may not work in another). Can the approaches developed in Australia play a useful role for controlling this invasive species in Japan? Our experimental trials on Ishigaki Island suggest that these new methods can be successfully applied to Japan. First, Cane Toad embryos exposed to chemical cues of conspecific tadpoles exhibited a reduction in viability (subsequent growth and development). This response appears to be species-specific, with native frog embryos not being affected by exposure to cues from toad tadpoles, and Cane Toad embryos not being affected by exposure to cues from native frog tadpoles. Second, Cane Toad tadpoles were attracted to traps containing water from conspecific eggs, and toxin from adult conspecifics. Third, adult Cane Toads were attracted to acoustic cues of calling males, with sex differences in rates of attraction to specific versions of a synthetic call (males were attracted to choruses whereas females were attracted to low-frequency calls). Our results suggest that the methods developed by Australian researchers are applicable to controlling invasive Cane Toads in Japan.


Innate inhibition of adaptive immunity: Mycobacterium tuberculosis-induced IL-6 inhibits macrophage responses to IFN-gamma.
In humans and in mice, control of the intracellular pathogen, Mycobacterium tuberculosis (Mtb), requires IFN-gamma. Although the adaptive immune response results in production of substantial amounts of IFN-gamma in response to Mtb, the immune response is unable to eradicate the infection in most cases. We have previously reported evidence that Mtb inhibits macrophage responses to IFN-gamma, suggesting that this may limit the ability of IFN-gamma to stimulate macrophages to kill Mtb. We have also observed that uninfected macrophages, adjacent to infected macrophages in culture, exhibit decreased responses to IFN-gamma. Here we report that IL-6 secreted by Mtb-infected macrophages inhibits the responses of uninfected macrophages to IFN-gamma. IL-6 selectively inhibits a subset of IFN-gamma-responsive genes at the level of transcriptional activation without inhibiting activation or function of STAT1. Inhibition of macrophage responses to IFN-gamma by IL-6 requires new protein synthesis, but this effect is not attributable to suppressor of cytokine signaling 1 or 3. These results reveal a novel function for IL-6 and indicate that IL-6 secreted by Mtb-infected macrophages may contribute to the inability of the cellular immune response to eradicate infection.


Is there any association between irritable bowel syndrome subgroups and autonomous dysfunction.
Irritable bowel syndrome (IBS) is a common functional intestinal disorder. Although there are marked improvements in the conceptualization of IBS pathophysiology in brain-intestinal interaction disorder, there is no definite consensus in the role of autonomic dysfunction (AD) in disease development and symptom progression. The aim of this study was to evaluate autonomous dysfunction in IBS subgroups. A total of 50 patients and 49 healthy controls were included. IBS subgroup types and demographic characteristics of patients were recorded. AD investigations were made up of parasympathetic and sympathetic tests. There was no statistically significant difference was determined between the groups in accordance to demographic characteristics such as age, gender, BMI and resting heart rate (p > 0.05). Three parasympathetic and one sympathetic autonomic neuropathy tests were found significantly different (respectively p < 0.001, p = 0.001, p = 0.016, p < 0.001, p = 0.375). There were significant decreases in parasympathetic tests in IBS-C patients; however, in the control group, there were significant decreases in sympathetic tests when compared with IBS-D patients (p < 0.001). The severity of AD in IBS-C subgroup was more pronounced than the IBS-D subgroup. No correlation was determined between dysautonomia and disease duration (p > 0.05). AD may have a role in IBS pathophysiology. Deterioration of the autonomous system not only affects the gastrointestinal system but also other systems including the cardiovascular system. Patients may also be susceptible to more diverse problems.


Parts of activities: reply to Fellbaum and Miller (1990).
If people believe that one activity is a kind of another, they also tend to believe that the second activity is a part of the first. For example, they assert that deciding is a kind of thinking and that thinking is a part of deciding. Fellbaum and Miller's (1990) explanation for this phenomenon is based on the idea that people interpret part of in the domain of verbs as a type of logical entailment. Their explanation, however, suffers from at least 2 deficiencies. First, it fails to account for parallel effects with nouns (e.g., a contest is a kind of an activity, and an activity is a part of a contest). Second, it contains a flaw that incorrectly predicts many activities to be parts of each other (e.g., coming is part of going and going part of coming). However, a hypothesis Rips and Conrad (1989) originally proposed for the kind-part reciprocal effect avoids both of these difficulties.


Previous medical problems in 326 consecutive hip fracture patients.
Pre-existing medical problems of elderly patients with hip fracture are seldom considered in orthopaedic literature, although they are indisputably the most important determinants for mortality, morbidity and final outcome. It is the purpose of this study to determine these problems in our hip fracture patients. Previous medical disorders and treatments, age, sex and type of fracture were prospectively recorded from all patients over 65 years old, diagnosed with hip fracture in a tertiary university general hospital during 2004. There were 326 patients who fractured their hip (81.04 hip fractures/100,000 people/year) (83.67.3 years old) (85.3% female). The patients existing medical conditions included hypertension (53% of patients), diabetes (19%), dementia (18%), cerebrovascular disease (11%), cataracts and/or blindness (10%), cardiac arrhythmia (9%), chronic obstructive pulmonary disease (9%), heart failure (8%), ischaemic heart disease (7%), psychiatric disorders other than dementia (7%), peptic ulcer (7%), and Parkinson's disease (5%); only 7% had no known significant medical problem beyond their fracture. Cardiovascular and neurological disorders, the most frequent, were also the most dangerous as potential sources for complications and difficulties during anaesthesia, surgery, immediate postoperative period and rehabilitation. Diabetes, the second most frequent diagnosis, complicated any other existing condition.


Perioperative Nutritional Management in Congenital Perineal and Vestibular Fistulas: A Systematic Review.
Surgical advancements have led to improved outcomes for children with congenital anorectal malformations with vestibular and perineal fistulas. However, the effect of perioperative nutritional management is debated and guidelines have not yet been established. The study aims to give an overview of available published evidence, regarding the impact of different perioperative nutritional management protocols on surgical outcome. A systematic literature review was conducted using PubMed, Embase, Cochrane Library, and CINAHL databases. All original articles concerning perioperative nutrition in children with vestibular and perineal fistulas were included. Methodological quality was assessed with the Rangel score. Included studies were subdivided into two groups: early enteral nutrition and prolonged fasting with or without parenteral nutrition. The database search resulted in 768 publications. Nine studies were eligible for inclusion. Wound complications were present in 56 of the 1,557 patients (4%) in whom this was assessed, and were more frequently seen in the prolonged fasting group (2 vs. 10%, p < 0.0001). Regarding the long-term outcome, constipation (grade II-III) was seen in 4% of the early feeding group, compared with 13% in the prolonged fasting group (p < 0.0001). This systematic review presents an overview of studies reporting on perioperative nutritional management in children with perineal and vestibular fistulas. Although study quality is low and study heterogeneity may also influence our results, early enteral feeding seems to be the preferable postoperative feeding strategy. Both early wound complications as well as long-term complications, in terms of clinically relevant constipation, seem to be lower in the early enteral feeding group. However, a prospective randomized, multicentered trial should be initiated to draw definitive conclusions regarding this matter.


Does diet-beverage intake affect dietary consumption patterns? Results from the Choose Healthy Options Consciously Everyday (CHOICE) randomized clinical trial.
Little is understood about the effect of increased consumption of low-calorie sweeteners in diet beverages on dietary patterns and energy intake. We investigated whether energy intakes and dietary patterns were different in subjects who were randomly assigned to substitute caloric beverages with either water or diet beverages (DBs). Participants from the Choose Healthy Options Consciously Everyday randomized clinical trial (a 6-mo, 3-arm study) were included in the analysis [water groups: n = 106 (94% women); DB group: n = 104 (82% women)]. For energy, macronutrient, and food and beverage intakes, we investigated the main effects of time, treatment, and the treatment-by-time interaction by using mixed models. Overall, the macronutrient composition changed in both groups without significant differences between groups over time. Both groups reduced absolute intakes of total daily energy, carbohydrates, fat, protein, saturated fat, total sugar, added sugar, and other carbohydrates. The DB group decreased energy from all beverages more than the water group did only at month 3 (P-group-by-time < 0.05). Although the water group had a greater reduction in grain intake at month 3 and a greater increase in fruit and vegetable intake at month 6 (P-group-by-time < 0.05), the DB group had a greater reduction in dessert intake than the water group did at month 6 (P-group-by-time < 0.05). Participants in both intervention groups showed positive changes in energy intakes and dietary patterns. The DB group showed decreases in most caloric beverages and specifically reduced more desserts than the water group did. Our study does not provide evidence to suggest that a short-term consumption of DBs, compared with water, increases preferences for sweet foods and beverages. This trial was registered at clinicaltrials.gov as NCT01017783.


miR-942 promotes proliferation and metastasis of hepatocellular carcinoma cells by inhibiting RRM2B.
Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related death. MicroRNA-942 (miR-942) plays a critical role in promoting proliferation and metastasis of cancer cells and is associated with poor prognosis in some types of cancers. However, the prognostic value of miR-942 and its functional role in HCC remain unclear. Real-time PCR (RT-PCR) was used to detect the expression of miR-942 in HCC tissues and adjacent normal liver tissues. Next, the correlations between miR-942 expression and clinicopathological parameters including the survival rate were analyzed. Interaction between miR-942 and ribonucleotide reductase regulatory TP53 inducible subunit M2B (RRM2B) was determined by RT-PCR, Western blot and luciferase assay. The biological influence of miR-942 on HCC cell lines was studied using CCK-8 assay, colony formation assay and transwell assay in vitro. Western blot and RT-PCR were used to analyze the change of downstream genes after miR-942 mimics transfection. miR-942 was significantly up-regulated in HCC. Its high expression was associated with serum alanine transaminase level (P=0.0350), tumor size (P=0.0195), T stage (P=0.0045) and lymphatic metastasis (P=0.0013). High expression of miR-942 was associated with shorter overall survival and disease-free survival time of HCC patients. RRM2B was validated as a target gene of miR-942. miR-942 mimics markedly promoted the malignant phenotypes of Huh7 and MHCC97H cell lines, while its inhibitor had the opposite effect. miR-942 can regulate the downstream genes of RRM2B including Egr-1 and PTEN, markers of epithelial-mesenchymal transition and matrix metalloproteinases. miR-942 may serve as a potential biomarker for HCC and its inhibitor may be a therapeutic agent for the treatment of this deadly disease.


Analysis of Electron Transparent Beam-Sensitive Samples Using Scanning Electron Microscopy Coupled With Energy-Dispersive X-ray Spectroscopy.
Scanning electron microscopy, coupled with energy-dispersive X-ray spectroscopy (EDS), is a powerful tool used in many scientific fields. It can provide nanoscale images, allowing size and morphology measurements, as well as provide information on the spatial distribution of elements in a sample. This study compares the capabilities of a traditional EDS detector with a recently developed annular EDS detector when analyzing electron transparent and beam-sensitive NaCl particles on a TEM grid. The optimal settings for single particle analysis are identified in order to minimize beam damage and optimize sample throughput via the choice of acceleration voltage, EDS acquisition time, and quantification model. Here, a linear combination of two models is used to bridge results for particle sizes, which are neither bulk nor sufficiently thin to assume electron transparent. Additionally, we show that the increased count rate obtainable with the annular detector enables mapping as a viable analysis strategy compared with feature detection methods, which only scan segmented regions. Finally, we discuss advantages and disadvantages of the two analysis strategies.


Spiromustine analogues. Relationships between structure, plasma stability, and antitumor activity.
Spiromustine is a hydantoin-containing nitrogen mustard currently in Phase I clinical trial. Since the in vitro plasma half-life of this compound (6.4 min, 37 degrees C, pH 7.4) appeared to be influenced by the hydantoin ring, analogues containing 2-5 methylene spacer groups between this ring and the nitrogen mustard moiety were prepared and evaluated for hydrolytic stability and antitumor activity. Stability correlated with structure and pKa values. The proximity of the hydantoin ring to the mustard function was a stabilizing factor. Activity against murine P-388 leukemia was demonstrated and a gradual decrease in this activity was observed as the hydrolytic instability increased. A relationship between analogue structure and a mass spectral rearrangement ion was identified.


Identification and deletion of Tft1, a predicted glycosyltransferase necessary for cell wall β-1,3;1,4-glucan synthesis in Aspergillus fumigatus.
Aspergillus fumigatus is an environmental mold that causes severe, often fatal invasive infections in immunocompromised patients. The search for new antifungal drug targets is critical, and the synthesis of the cell wall represents a potential area to find such a target. Embedded within the main β-1,3-glucan core of the A. fumigatus cell wall is a mixed linkage, β-D-(1,3;1,4)-glucan. The role of this molecule or how it is synthesized is unknown, though it comprises 10% of the glucans within the wall. While this is not a well-studied molecule in fungi, it has been studied in plants. Using the sequences of two plant mixed linkage glucan synthases, a single ortholog was identified in A. fumigatus (Tft1). A strain lacking this enzyme (tft1Δ) was generated along with revertant strains containing the native gene under the control of either the native or a strongly expressing promoter. Immunofluorescence staining with an antibody against β-(1,3;1,4)-glucan and biochemical quantification of this polysaccharide in the tft1Δ strain demonstrated complete loss of this molecule. Reintroduction of the gene into the knockout strain yielded reappearance in amounts that correlated with expected expression of the gene. The loss of Tft1 and mixed linkage glucan yielded no in vitro growth phenotype. However, there was a modest increase in virulence for the tft1Δ strain in a wax worm model. While the precise roles for β-(1,3;1,4)-glucan within A. fumigatus cell wall are still uncertain, it is clear that Tft1 plays a pivotal role in the biosynthesis of this cell wall polysaccharide.


Buccal spectral markers for lung cancer risk stratification.
Lung cancer remains the leading cause of cancer deaths in the US with >150,000 deaths per year. In order to more effectively reduce lung cancer mortality, more sophisticated screening paradigms are needed. Previously, our group demonstrated the use of low-coherence enhanced backscattering (LEBS) spectroscopy to detect and quantify the micro/nano-architectural correlates of colorectal and pancreatic field carcinogenesis. In the lung, the buccal (cheek) mucosa has been suggested as an excellent surrogate site in the "field of injury". We, therefore, wanted to assess whether LEBS could similarly sense the presence of lung. To this end, we applied a fiber-optic LEBS probe to a dataset of 27 smokers without diagnosed lung cancer (controls) and 46 with lung cancer (cases), which was divided into a training and a blinded validation set (32 and 41 subjects, respectively). LEBS readings of the buccal mucosa were taken from the oral cavity applying gentle contact. The diagnostic LEBS marker was notably altered in patients harboring lung cancer compared to smoking controls. The prediction rule developed on training set data provided excellent diagnostics with 94% sensitivity, 80% specificity, and 95% accuracy. Applying the same threshold to the blinded validation set yielded 79% sensitivity and 83% specificity. These results were not confounded by patient demographics or impacted by cancer type or location. Moreover, the prediction rule was robust across all stages of cancer including stage I. We envision the use of LEBS as the first part of a two-step paradigm shift in lung cancer screening in which patients with high LEBS risk markers are funnelled into more invasive screening for confirmation.


Experimental and computational data set on adsorption of Cr (VI) from water using an activated carbon.
Chromium (Cr) is a widely used metal in metallurgical and chemical industries, whose waste contaminates the surface and groundwater. Cr (VI) is toxic and produces carcinogenic effects owing to its high mobility in water and soil. In this work, computational and experimental studies from the adsorption of Cr(VI) from aqueous solutions on teak wood residues activated with ZnCl2 (AT) are presented. Full interpretation of data can be found in DOI:10.1016/j.jece.2020.103702 [1]. Experimental data were adjusted to Langmuir, Freundlich and Temkin isothermal models and the nonlinear and linear forms of the Pseudo-first and Pseudo-second order kinetic models. Computational data allow to understand the adsorption process of Cr(VI) on carbonaceous materials.


Structure of Drosophila polytene chromosomes. Evidence for a toroidal organization of the bands.
Avoiding acid fixation or squashing, the structure of Drosophila salivary gland polytene chromosomes has been examined in detail in nuclei, with special emphasis on the organization of the DNA in the chromosome bands. Cut serial sections, optical serial sections, scanning electron microscopy (SEM) on whole mounts, high-voltage electron microscopy (HVEM) on whole mounts, and pancreatic DNase I digestion monitored by fluorescent microscopy have been used to complement one another in this analysis. With all five of these techniques, stereo pairs were used to aid in the three-dimensional reconstruction of chromosomal structures. Evidence is presented that most, if not all, of the polytene chromosome bands are torus-shaped. The DNA of these bands is largely confined to the rim, with the interior essentially DNA-free. The chromatin in each polytene band is also seen to have an extremely regular and highly ordered substructure. This substructural organization is largely radially symmetric in the bands and generally parallel to the chromosome axis. In addition, each band appears to be a distinct architectural entity with regard to its exact structural features and dimensions. A model is presented that follows these organizational boundary conditions.


Mechanisms of subcutaneous allergen immunotherapy.
Allergen-specific immunotherapy (SIT) is the only curative approach in the treatment of allergic diseases defined up-to-date. Peripheral T-cell tolerance to allergens, the goal of successful allergen-SIT, is the primary mechanism in healthy immune responses to allergens. By repeated administration of increased doses of the causative allergen, allergen-SIT induces a state of immune tolerance to allergens through the constitution of T regulatory (Treg) cells, including allergen-specific interleukin (IL)-10-secreting Treg type 1 cells and CD4(+)CD25(+)Treg cells; induction of suppressive cytokines, such as IL-10 and transforming growth factor β; suppression of allergen-specific IgE and induction of IgG4 and IgA; and suppression of mast cells, basophils, eosinophils, and inflammatory dendritic cells. This review summarizes the current knowledge on the mechanisms of allergen-SIT with emphasis on the roles of Treg cells in allergen-SIT.


Machine learning identification of Pseudomonas aeruginosa strains from colony image data.
When grown on agar surfaces, microbes can produce distinct multicellular spatial structures called colonies, which contain characteristic sizes, shapes, edges, textures, and degrees of opacity and color. For over one hundred years, researchers have used these morphology cues to classify bacteria and guide more targeted treatment of pathogens. Advances in genome sequencing technology have revolutionized our ability to classify bacterial isolates and while genomic methods are in the ascendancy, morphological characterization of bacterial species has made a resurgence due to increased computing capacities and widespread application of machine learning tools. In this paper, we revisit the topic of colony morphotype on the within-species scale and apply concepts from image processing, computer vision, and deep learning to a dataset of 69 environmental and clinical Pseudomonas aeruginosa strains. We find that colony morphology and complexity under common laboratory conditions is a robust, repeatable phenotype on the level of individual strains, and therefore forms a potential basis for strain classification. We then use a deep convolutional neural network approach with a combination of data augmentation and transfer learning to overcome the typical data starvation problem in biological applications of deep learning. Using a train/validation/test split, our results achieve an average validation accuracy of 92.9% and an average test accuracy of 90.7% for the classification of individual strains. These results indicate that bacterial strains have characteristic visual 'fingerprints' that can serve as the basis of classification on a sub-species level. Our work illustrates the potential of image-based classification of bacterial pathogens and highlights the potential to use similar approaches to predict medically relevant strain characteristics like antibiotic resistance and virulence from colony data.


Cholecystectomy concomitant with laparoscopic gastric bypass: a trend analysis of the nationwide inpatient sample from 2001 to 2008.
Gallstone formation is common in obese patients, particularly during rapid weight loss. Whether a concomitant cholecystectomy should be performed during laparoscopic gastric bypass surgery is still contentious. We aimed to analyze trends in concomitant cholecystectomy and laparoscopic gastric bypass surgery (2001-2008), to identify factors associated with concomitant cholecystectomy, and to compare short-term outcomes after laparoscopic gastric bypass with and without concomitant cholecystectomy. We used data from adults undergoing laparoscopic gastric bypass for obesity from the Nationwide Inpatient Sample. The Cochran-Armitage trend test was used to assess changes over time. Unadjusted and risk-adjusted generalized linear models were performed to assess predictors of concomitant cholecystectomy and to assess postoperative short-term outcomes. A total of 70,287 patients were included: mean age was 43.1 years and 81.6% were female. Concomitant cholecystectomy was performed in 6,402 (9.1%) patients. The proportion of patients undergoing concomitant cholecystectomy decreased significantly from 26.3% in 2001 to 3.7% in 2008 (p for trend < 0.001). Patients who underwent concomitant cholecystectomy had higher rates of mortality (unadjusted odds ratios [OR], 2.16; p = 0.012), overall postoperative complications (risk-adjusted OR, 1.59; p = 0.001), and reinterventions (risk-adjusted OR, 3.83; p < 0.001), less frequent routine discharge (risk-adjusted OR, 0.70; p = 0.05), and longer adjusted hospital stay (median difference, 0.4 days; p < 0.001). Concomitant cholecystectomy and laparoscopic gastric bypass surgery have decreased significantly over the last decade. Given the higher rates of postoperative complications, reinterventions, mortality, as well as longer hospital stay, concomitant cholecystectomy should only be considered in patients with symptomatic gallbladder disease.


A confocal and electron microscopic comparison of interferon beta-induced changes in vesicular stomatitis virus infection of neuroblastoma and nonneuronal cells.
Vesicular stomatitis virus (VSV) replication is highly sensitive to interferon (IFN)-induced antiviral responses. Pretreatment of sensitive cultured cells with IFNbeta results in a 10(4)-fold reduction in the release of infectious VSV particles. However, differences exist between the mechanisms of reduced infectious particle titers in cell lines of neuroblastoma and nonneuronal lineage. In L929-fibroblast-derived cells, using immunofluorescence confocal microscopy, infection under control conditions reveals the accumulation of VSV matrix, phosphoprotein (P), and nucleocapsid (N) proteins over time, with induced cellular morphological changes indicative of cytopathic effects (CPEs). Upon observing L929 cells that had been pretreated with IFNbeta, neither detectable VSV proteins nor CPEs were seen, consistent with type I IFN antiviral protection. When using the same techniques to observe VSV infections of NB41A3 cells, a neuroblastoma cell line, aside from similar viral progression in the untreated control cells, IFNbeta-treated cells illustrated a severely attenuated VSV infection. Attenuated VSV progression was observed through detection of VSV matrix, P, and N proteins in isolated cells during the first 8 h of infection. However, by 18-24 h postinfection all neuroblastomas had succumbed to the viral infection. Finally, upon closer inspection of IFNbeta-treated NB41A3 cells, no detectable changes in VSV protein localization were identified compared with untreated, virally infected neuroblastomas. Next, to extend our study to test our hypothesis that virion assembly is compromised within type I IFN-treated neuroblastoma cells, we employed electron microscopy to examine our experimental conditions at the ultrastructural level. Using VSV-specific antibodies in conjunction with immuno-gold reagents, we observed several similarities between the two cell lines, such as identification of viroplasmic regions containing VSV N and P proteins and signs of stress-induced CPEs of VSV-infected cells, which had either been mock-treated or pretreated with interferon-beta (IFNbeta). One difference we observed between nonneuronal and neuroblastoma cells was more numerous actively budding VSV virions across untreated L929 plasma membranes compared with untreated NB41A3 cells. Additionally, IFNbeta-treated, VSV-infected L929 cells exhibited neither cytoplasmic viroplasm nor viral protein expression. In contrast, IFNbeta-treated, VSV-infected NB41A3 cells showed evidence of VSV infection at a very low frequency as well as small-scale viroplasmic regions that colocalized with viral N and P proteins. Finally, we observed that VSV viral particles harvested from untreated VSV-infected L929 and NB41A3 cells were statistically similar in size and shape. A portion of VSV virions from IFNbeta-treated, virally infected NB41A3 cells were similar in size and shape to virus from both untreated cell types. However, among the sampling of virions, pleomorphic viral particles that were identified from IFNbeta-treated, VSV-infected NB41A3 cells were different enough to suggest a misassembly mechanism as part of the IFNbeta antiviral state in neuroblastoma cells.


Etiologies, Trends, and Predictors of 30-Day Readmission in Patients With Heart Failure.
Heart failure (HF) is the most common discharge diagnosis across the United States, and these patients are particularly vulnerable to readmissions, increasing attention to potential ways to address the problem. The study cohort was derived from the Healthcare Cost and Utilization Project's National Readmission Data 2013, sponsored by the Agency for Healthcare Research and Quality. HF was identified using appropriate International Classification of Diseases, Ninth Revision, Clinical Modification codes. Readmission was defined as a subsequent hospital admission within 30 days after discharge day of index admission. Readmission causes were identified using International Classification of Diseases, Ninth Revision, codes in primary diagnosis filed. The primary outcome was 30-day readmission. Hierarchical 2-level logistic models were used to evaluate study outcomes. From a total 301,892 principal admissions (73.4% age ≥65 years and 50.6% men), 55,857 (18.5%) patients were readmitted with a total of 64,264 readmissions during the study year. Among the etiologies of readmission, cardiac causes (49.8%) were most common (HF being most common followed by coronary artery disease and arrhythmias), whereas pulmonary causes were responsible for 13.1% and renal causes for 8.9% of the readmissions. Significant predictors of increased 30-day readmission included diabetes (odds ratio, 95% confidence interval, p value: 1.06, 1.03 to 1.08, p <0.001), chronic lung disease (1.13, 1.11 to 1.16, p <0.001), renal failure/electrolyte imbalance (1.12, 1.10 to 1.15, p <0.001), discharge to facilities (1.07, 1.04 to 1.09, p <0.001), lengthier hospital stay, and transfusion during index admission. In conclusion, readmission after a hospitalization for HF is common. Although it may be necessary to readmit some patients, the striking rate of readmission demands efforts to further clarify the determinants of readmission and develop strategies in terms of quality of care and care transitions to prevent this adverse outcome.


Degradation of endocrine disrupting chemicals in aqueous solution by interaction of photocatalytic oxidation and ferrate (VI) oxidation.
In this study, the degradation of bisphenol A in aqueous suspension by interaction of photocatalytic oxidation and ferrate(VI) oxidation was investigated under different conditions. The results indicate that the formation of Fe(V) and Fe(IV) is in the photocatalytic reduction of Fe(VI) by electron (ecb-) on the surface of TiO2. The oxidation efficiency of the photocatalytic oxidation in the presence of Fe(VI) was much greater than that without. In addition, the decomposition of Fe(VI) under different conditions was also investigated. The results indicate that the Fe(VI) reduction was accelerated by photocatalytic reaction and the adsorption capacity of Fe(VI) on TiO2 surface decreased as pH increased. The characteristics of solid potassium ferrate prepared were investigated by X-ray diffraction. It was found that the potassium ferrate solid has a tetrahedral structure with a space group of D2h (Pnma) and a = 7.705 A, b = 5.863 A, and c = 10.36 A.


Percutaneous transluminal coronary angioplasty in 1985-1986 and 1977-1981. The National Heart, Lung, and Blood Institute Registry.
In August 1985, the Percutaneous Transluminal Coronary Angioplasty Registry of the National Heart, Lung, and Blood Institute reopened at its previous sites to document changes in angioplasty strategy and outcome. The new registry entered 1802 consecutive patients who had not had a myocardial infarction in the 10 days before angioplasty. Patient selection, technical outcome, and short-term major complications were compared with those of the 1977 to 1981 registry cohort. The new-registry patients were older and had a significantly higher proportion of multivessel disease (53 vs. 25 percent, P less than 0.001), poor left ventricular function (19 vs. 8 percent, P less than 0.001), previous myocardial infarction (37 vs. 21 percent, P less than 0.001), and previous coronary bypass surgery (13 vs. 9 percent, P less than 0.01). The new-registry cohort also had more complex coronary lesions, and angioplasty attempts in these patients involved more multivessel procedures. Despite these differences, the in-hospital outcome in the new cohort was better. Angiographic success rates according to lesion increased from 67 to 88 percent (P less than 0.001), and overall success rates (measured as a reduction of at least 20 percent in all lesions attempted, without death, myocardial infarction, or coronary bypass surgery) increased from 61 to 78 percent (P less than 0.001). In-hospital mortality for the new cohort was 1 percent, and the nonfatal myocardial infarction rate was 4.3 percent. Both rates are similar to those for the old registry. The long-term efficacy of current angioplasty remains to be determined.


Detection of the nifH gene of Methanobrevibacter smithii: a potential tool to identify sewage pollution in recreational waters.
The goal of this study was to develop and test the efficacy of a PCR assay for the environmental detection of the nifH gene of Methanobrevibacter smithii, a methanogen found in human faeces and sewage. PCR primers for the nifH gene of M. smithii were designed, tested and used to detect the presence or absence of this organism in faecal and environmental samples. Specificity analysis showed that the Mnif primers amplified products only in M. smithii pure culture strains (100%), human faeces (29%), human sewage samples (93%) and sewage-contaminated water samples (100%). No amplification was observed when primers were tested against 43 bacterial stock cultures, 204 animal faecal samples, 548 environmental bacterial isolates and water samples from a bovine waste lagoon and adjacent polluted creek. Sequencing of PCR products from sewers demonstrated that a 222-bp product was the nifH gene of M. smithii. The minimal amount of total DNA required for the detection of M. smithii was 10 ng for human faeces, 10 ng for faecally contaminated water and 5 ng for sewage. Recreational water seeded with M. smithii established a lower detection limit of 13 cells ml(-1). The Mnif assay developed during this investigation showed successful detection of M. smithii in individual human faecal samples, sewage and sewage-contaminated water but not in uncontaminated marine water or bovine-contaminated waters. The Mnif assay appears to be a potentially useful method to detect sewage-polluted coastal waters. This study was the first to utilize methanogens as an indicator of sewage pollution. Mnif PCR detection of M. smithii was shown to be a rapid, inexpensive and reliable test for determining the presence or absence of sewage pollution in coastal recreational waters.


Control of lower esophageal sphincter pressure by two sites in dorsal motor nucleus of the vagus.
Our purpose was to determine the central vagal sites for regulating changes in lower esophageal sphincter (LES) pressure in the cat. Injection of the retrograde tracer, horseradish peroxidase, into the LES resulted in labeling of cells in the dorsal motor nucleus of the vagus (DMV), with the largest number of cells appearing in two areas, one area rostral to obex (1.5-4.0 mm) and one area caudal to obex (-0.5 to -1.5 mm). In alpha-chloralose-anesthetized cats, L-glutamic acid was microinjected into these areas and LES pressure, intragastric pressure, and stomach motility were monitored. Microinjection of L-glutamic acid into the rostral area resulted in significant increases in LES pressure (18.6 +/- 4.9 mmHg; P less than 0.05), pyloric motility (baseline minute motility increased from 5.7 +/- 2.2 to 14.5 +/- 3.9 postinjection; P less than 0.05) and stomach pressure (baseline of 16.9 +/- 2.3 mmHg increased to 23.8 +/- 3.7 mmHg postinjection; P less than 0.05). Microinjection of L-glutamic acid into the caudal area resulted in significant decreases in LES pressure (-14.3 +/- 5.8 mmHg; P less than 0.05) and intragastric pressure (-7.5 +/- 2.2 mmHg; P less than 0.05) with no significant changes in pyloric motility. Ipsilateral vagotomy abolished both sets of responses. These data indicate that excitatory and inhibitory control of LES and intragastric pressure are mediated by vagal efferent neurons located in two distinct sites in the DMV.


Immune response to an epitope of the NS4 protein of hepatitis C virus in HCV-related disorders.
NS4, a nonstructural protein of HCV, is a frequent target of antibodies in infected subjects. According to recent data, the antibodies frequently recognize the sequence 1921-40 of the NS4 protein. The aim of this work was to analyze antibody reactivity with the sequence 1921-40 in different HCV-related disorders. Although this sequence is located in a relatively invariant region of viral genome, two strain-specific sequences are described. Thus, three NS4 1921-1940 peptides were synthesized: the BK shared by most viral strains, the J6 (strain 2a), and the J8 (strain 2b). The peptides were used as antigens in the solid phase for measuring serum IgG antibodies in an ELISA assay. Antibodies reactive with the 1921-40 BK peptide were detected in 64% of sera from patients with autoimmune hepatitis (AIH), 51% from chronic hepatitis C (CHC), and 22% from mixed cryoglobulinemia (MC). The frequency of positive sera in MC was significatively lower than in AIH (P < 0.0001) or CHC (P < 0.0021). Similar results were obtained with the J6 and J8 peptides. All sera that did not react with the BK peptide were negative on J6 and J8 and conversely most sera reacting with the BK peptide also bound the J6 and the J8 peptides. No correlation was found between the genotype of the infecting virus and the presence of antibodies to any of the NS4 peptides. These results indicate that many HCV-infected subjects produce antibodies to the NS4 sequence 1921-40. The immune response to this sequence is not strain specific and varies with the different disorders associated with HCV infection.


Definitions of genital ulcer disease and variation in risk for prevalent human immunodeficiency virus infection.
Although genital ulcer disease (GUD) has been associated with human immunodeficiency virus (HIV) infection in a number of studies, definitions of genital ulceration have varied. The authors hypothesized that the association of GUD with prevalent HIV infection may vary according to the definition of GUD that is used. As part of a prospective cohort study, 863 patients were interviewed and examined who presented to a sexually transmitted disease (STD) clinic for new symptom evaluation and who agreed to HIV testing to determine demographic and behavioral risk associated with prevalent HIV infection. To determine the association between GUD and prevalent HIV, the following definitions of GUD were used: observed ulcers, history of syphilis, serologic evidence of syphilis, observed culture-proven genital herpes, and serologic evidence of herpes simplex virus type II (HSV-2) infection. Of 481 men and 382 women enrolled, prevalent HIV infection was detected in 12.5% and 5.2%, respectively. In multivariate analyses controlling for known HIV risk behaviors, prevalent HIV infection was associated with observed GUD (odds ratio [OR] = 2.0, 95% confidence intervals (CI) = 1.0-3.9), a history of syphilis (OR = 6.0, CI = 2.8-12.7), and serologic evidence of syphilis (OR = 3.7, CI = 1.9-7.0), but not with serologic evidence of HSV-2 (OR = 1.2, CI = 0.7-2.1), nor with observed HSV-2 culture-positive genital ulcerations (OR = 1.0, CI = 0.4-4.2). Factors contributing to different strengths of association between HIV infection and a history of syphilis or serologic evidence of syphilis included the presence of underdiagnosed syphilis infection in people with reactive serologic tests and the absence of serologic reactivity in people with a positive history. Although GUD is strongly associated with prevalent HIV, the strength of the association depends on the definition of GUD used. For accurate evaluation of people at risk for HIV, clinicians and researchers should use multiple definitions of GUD.


Mammographic features and histopathological findings of interval breast cancers.
Interval cancers are considered a shortcoming in screening mammography due to less favorable prognostic tumor characteristics compared to screening-detected cancers and consequently a lower chance of survival from the disease. To describe the mammographic features and prognostic histopathological tumor characteristics of interval breast cancers. A total of 231 interval breast cancer cases diagnosed in prevalently screened women aged 50-69 years old were examined. Thirty-five percent of the cases were retrospectively classified as missed cancers, 23% as minimal sign, and 42% as true negative (including occult cancers) in a definitive classification performed by six experienced breast radiologists. The retrospective classification described the mammographic features of the baseline screening mammograms in missed and minimal-sign interval cancers, while histopathological reports were used to describe the tumor characteristics in all the subgroups of interval cancers. Fifty percent of the missed and minimal-sign interval cancers combined presented poorly defined mass or asymmetric density, and 26% calcifications with or without associated density or mass at baseline screening. Twenty-seven percent of invasive tumors were <15 mm for missed and 47% for true interval cancers (P<0.001). Lymph node involvement was more common in missed (49%) compared with the true cases (33%, P<0.05). Missed interval cancers have less prognostically favorable histopathological tumor characteristics compared with true interval cancers. Improving the radiologist's perception and interpretation by establishing systematic collection of features and implementation of organized reviews may decrease the number of interval cancers in a screening program.


Symptoms of sleep disorders, inattention, and hyperactivity in children.
Children with sleep disorders are often inattentive or hyperactive, and some carry a diagnosis of attention deficit/hyperactivity disorder (ADHD) until their sleep disorder is detected. However, the potential behavioral impact of undiagnosed sleep disorders is not known. We sought to determine whether children with higher levels of inattention and hyperactivity more frequently have symptoms of sleep-related breathing disorders (SRBDs) or periodic limb movement disorder (PLMD). We surveyed parents of 2-18-year-old patients at a child psychiatry clinic (n = 70) and a general pediatrics clinic (n = 73) to assess the children's behavior, snoring, complaints of restless legs at night, and daytime sleepiness. A validated pediatric sleep questionnaire provided the explanatory variables, and a scale for inattention and hyperactivity, derived from the Diagnostic and Statistical Manual of Mental Disorders, 4th edition (DSM-IV), provided the dependent variable. Habitual snoring was more frequent (33%) among children who carried a diagnosis of ADHD than among the other children at the psychiatry or general pediatric clinics (11 and 9%, respectively, chi-square test, p = 0.01). Snoring scores, derived from six snoring- and SRBD-related question items, were associated with higher levels of inattention and hyperactivity. The complaint of restless legs and a composite score for daytime sleepiness showed some evidence, though less consistent, of an association with inattention and hyperactivity. The association of snoring with inattention and hyperactivity suggests that SRBDs and perhaps other sleep disorders could be a cause of inattention and hyperactivity in some children. If a causal effect is present, our data suggest that 81% of habitually snoring children who have ADHD--25% of all children with ADHD--could have their ADHD eliminated if their habitual snoring and any associated SRBD were effectively treated.


Ultrafast optics. Ultrafast optical control by few photons in engineered fiber.
Fast control of a strong optical beam by a few photons is an outstanding challenge that limits the performance of quantum sensors and optical processing devices. We report that a fast and efficient optical gate can be realized in an optical fiber that has been engineered with molecular-scale accuracy. Highly efficient, distributed phase-matched photon-photon interaction was achieved in the fiber with locally controlled, nanometer-scale core variations. A three-photon input was used to manipulate a Watt-scale beam at a speed exceeding 500 gigahertz. In addition to very fast beam control, the results provide a path to developing a new class of sensitive receivers capable of operating at very high rates.


Isolation of a polypeptide that has lymphocyte-differentiating properties and is probably represented universally in living cells.
A polypeptide of 8500 molecular weight is described that induces the differentiation of T (thymus-derived) cell and B (bone-marrow-derived) cell immunocytes in vitro, apparently via beta-adrenergic receptors and adenylate cyclase activation. This polypeptide shows a high degree of evolutionary conservation, exhibiting close structural, functional, and immunological similarity when isolated from such diverse origins as cells of mammals and higher plants. This polypeptide was detected in animal cells, yeast, bacteria, and higher plants, and so may well be a universal constituent of living cells.


Origin of 1/f PM and AM noise in bipolar junction transistor amplifiers.
In this paper we report the results of extensive research on phase modulation (PM) and amplitude modulation (AM) noise in linear bipolar junction transistor (BJT) amplifiers. BJT amplifiers exhibit 1/f PM and AM noise about a carrier signal that is much larger than the amplifiers thermal noise at those frequencies in the absence of the carrier signal. Our work shows that the 1/f PM noise of a BJT based amplifier is accompanied by 1/f AM noise which can be higher, lower, or nearly equal, depending on the circuit implementation. The 1/f AM and PM noise in BJTs is primarily the result of 1/f fluctuations in transistor current, transistor capacitance, circuit supply voltages, circuit impedances, and circuit configuration. We discuss the theory and present experimental data in reference to common emitter amplifiers, but the analysis can be applied to other configurations as well. This study provides the functional dependence of 1/f AM and PM noise on transistor parameters, circuit parameters, and signal frequency, thereby laying the groundwork for a comprehensive theory of 1/f AM and PM noise in BJT amplifiers. We show that in many cases the 1/f PM and AM noise can be reduced below the thermal noise of the amplifier.