Activity of cefiderocol (S-649266) against carbapenem-resistant Gram-negative bacteria collected from inpatients in Greek hospitals.
Cefiderocol (S-649266), a siderophore cephalosporin, utilizes a novel mechanism of entry into the periplasmic space of Gram-negative bacteria and is broadly stable to ESBLs and carbapenemases. A collection of carbapenem-resistant Gram-negative bacteria isolated from clinical specimens in 18 Greek hospitals was tested for susceptibility to cefiderocol, meropenem, ceftazidime, cefepime, ceftazidime/avibactam, ceftolozane/tazobactam, aztreonam, amikacin, ciprofloxacin, colistin and tigecycline. Broth microdilution plates were used to determine MICs. In total 189 non-fermentative Gram-negative bacteria (107 Acinetobacter baumannii and 82 Pseudomonas aeruginosa ) and 282 Enterobacteriaceae (including 244 Klebsiella pneumoniae , 14 Enterobacter cloacae and 11 Providencia stuartii ) were studied. For both A. baumannii and P. aeruginosa the MIC 90 of cefiderocol was 0.5 mg/L. For K. pneumoniae , E. cloacae and P. stuartii the MIC 90 of cefiderocol was 1, 1 and 0.5 mg/L, respectively. Tigecycline was the second most active antibiotic, followed by colistin. Cefiderocol exhibited greater antimicrobial activity in vitro against carbapenem-resistant Gram-negative bacteria than comparator antibiotics.


State-level clustering of safety measures and its relationship to injury mortality.
This article proposes a social model of investigating injury mortality. The authors hypothesize that (1) state-level laws and regulations on safety cluster together in one or more groupings; (2) groupings of safety measures play a significant role in injury mortality; and (3) injury mortality is very highly associated with social structural variables. There is a clustering of safety policies, with five factors explaining 67 percent of variance, although no "master factor" was discovered. The strongest factor, explaining 21 percent of variance, includes three gun laws and low speed limits before the 1973 federal law. One factor is the most global in that it taps three distinct areas, including helmet laws, minor blood alcohol levels, and smoke detectors, though it only explains 7.5 percent of variance. The only factor that remains in a regression for injury mortality is one that includes strong seat belt laws and strong enforcement of those laws, though in the direction opposite to that hypothesized. This factor, along with percentage rural and environmental spending per capita, is significant for both motor vehicle and non-motor vehicle mortality. For motor vehicle mortality alone, deaths are higher in states with higher percentages of Hispanics and fewer people receiving food stamps and AFDC. Many factors that usually predict individual injury mortality do not hold at the state level, suggesting the usefulness of looking at social factors for new insights into injury mortality and prevention.


Comparison of in vivo visual and computer-aided tooth shade determination.
The aim of this study was to evaluate the performance of shade-determining devices. For the daily practitioner, it is essential to know whether modern computer-aided shade selection is reliable in everyday life. So the question of how the clinical usability of these machines could be rated has to be clarified. In the following, three actual devices available in the market were compared using a human observer's perception. The SpectroShade device (MHT Optic Research AG, 8155 Niederhasli, Switzerland), the ShadeVision device (X-Rite Co., Grandville, USA) and the Digital Shade Guide DSG4 (A. Rieth, 73614 Schorndorf, Germany) were assessed with respect to their agreement with the color perception of three examiners looking at 57 test persons (six teeth each for a total of 342). Shades were reported in Vita Classical shades. It could be demonstrated that every single human examiner showed a significantly higher agreement value (human group on average 40.2%) when compared with the remaining five methods than each computer-aided tooth shade determination device. The devices reached on average only a value of 28.6%, whereas the X-Rite ShadeVision showed a significant better result (33.2%) than the MHT SpectroShade and Rieth DSG4 (27.0 and 25.7%). Identical shade results given by all three methods of a group (group of three devices and three humans) were found to be rather low for the computer-aided devices (9.9%) compared with humans (36.7%). All six methods together agreed in 3.3% of the cases. It becomes evident that the methods-especially the computer-aided shade determination-are rather divided about the respective tooth color. Deficiencies of the instrumental as well as the visual detection become obvious. The best agreement level was performed by the human examiners. The best agreement of the evaluated devices was obtained-generally as well as among the human testers-by the X-Rite ShadeVision system, followed at a statistically significant distance by the MHT SpectroShade and the Rieth DSG4. The agreement among the examiner group was 52.9%, significantly better than that of each device compared to this group (31.3% on average). Color detection and its realization are very complex. As shown, in many cases, computer-aided color shade determination of natural teeth seems to not reflect human perception.


Technical assessment of 2D and 3D imaging performance of an IGZO-based flat-panel X-ray detector.
Indirect detection flat-panel detectors (FPDs) consisting of hydrogenated amorphous silicon (a-Si:H) thin-film transistors (TFTs) are a prevalent technology for digital x-ray imaging. However, their performance is challenged in applications requiring low exposure levels, high spatial resolution, and high frame rate. Emerging FPD designs using metal oxide TFTs may offer potential performance improvements compared to FPDs based on a-Si:H TFTs. This work investigates the imaging performance of a new indium gallium zinc oxide (IGZO) TFT-based detector in 2D fluoroscopy and 3D cone-beam CT (CBCT). The new FPD consists of a sensor array combining IGZO TFTs with a-Si:H photodiodes and a 0.7-mm thick CsI:Tl scintillator. The FPD was implemented on an x-ray imaging bench with system geometry emulating intraoperative CBCT. A conventional FPD with a-Si:H TFTs and a 0.6-mm thick CsI:Tl scintillator was similarly implemented as a basis of comparison. 2D imaging performance was characterized in terms of electronic noise, sensitivity, linearity, lag, spatial resolution (modulation transfer function, MTF), image noise (noise-power spectrum, NPS), and detective quantum efficiency (DQE) with entrance air kerma (EAK) ranging from 0.3 to 1.2 μGy. 3D imaging performance was evaluated in terms of the 3D MTF and noise-equivalent quanta (NEQ), soft-tissue contrast-to-noise ratio (CNR), and image quality evident in anthropomorphic phantoms for a range of anatomical sites and dose, with weighted air kerma, K w ${K_w}$ , ranging from 0.8 to 4.9 mGy. The 2D imaging performance of the IGZO-based FPD exhibited up to ∼1.7× lower electronic noise than the a-Si:H FPD at matched pixel pitch. Furthermore, the IGZO FPD exhibited ∼27% increase in mid-frequency DQE (1 mm-1 ) at matched pixel size and dose (EAK ≈ 1.0 μGy) and ∼11% increase after adjusting for differences in scintillator thickness. 2D spatial resolution was limited by the scintillator for each FPD. The IGZO-based FPD demonstrated improved 3D NEQ at all spatial frequencies in both head (≥25% increase for all dose levels) and body (≥10% increase for K w ${K_w}$ ≤2 mGy) imaging scenarios. These characteristics translated to improved low-contrast visualization in anthropomorphic phantoms, demonstrating ≥10% improvement in CNR and extension of the low-dose range for which the detector is input-quantum limited. The IGZO-based FPD demonstrated improvements in electronic noise, image lag, and NEQ that translated to measurable improvements in 2D and 3D imaging performance compared to a conventional FPD based on a-Si:H TFTs. The improvements are most beneficial for 2D or 3D imaging scenarios involving low-dose and/or high-frame rate.


The existence of generalized synchronization of chaotic systems in complex networks.
The paper studies the existence of generalized synchronization in complex networks, which consist of chaotic systems. When a part of modified nodes are chaotic, and the others have asymptotically stable equilibriums or orbital asymptotically stable periodic solutions, under certain conditions, the existence of generalized synchronization can be turned to the problem of contractive fixed point in the family of Lipschitz functions. In addition, theoretical proofs are proposed to the exponential attractive property of generalized synchronization manifold. Numerical simulations validate the theory.


Tetrodotoxin in gastropods (snails) implicated in food poisoning in Northern Taiwan.
The toxin in the gastropods (snails) Zeuxis sufflatus and Niotha clathrata implicated in a food poisoning incident in northern Taiwan in April 2001 was studied. The symptoms exhibited by four victims were general paresthesia, paralysis of the phalanges and the extremities, paralysis, coma, vomiting, and aphasia. The remaining gastropods were assayed for toxicity in the form of tetrodotoxin (TTX). The ranges of specimen toxicity were 345 to 1,640 mouse units (MU) for Z sufflatus and 190 to 643 MU for N. clathrata. The toxicities of the digestive gland and for other parts of the gastropod were 1,120 +/- 477 MU and 497 +/- 238 MU, respectively, for Z sufflatus and 683 +/- 113 MU and 289 +/- 169 MU, respectively, for N. clathrata. The toxin from the methanolic extract of the gastropods was partially purified by ultrafiltration and Bio-Gel P-2 column chromatography. Cellulose acetate membrane electrophoresis, thin-layer chromatography, high-performance liquid chromatography, and gas chromatography-mass spectrometry analyses demonstrated that the toxin consisted of TTX. It was concluded that the causative agent of the food poisoning in question was TTX.


Results From the NACC Uniform Data Set Neuropsychological Battery Crosswalk Study.
Four new nonproprietary tests were recommended for use in the National Alzheimer's Coordinating Center's Uniform Data Set Neuropsychological Battery. These tests are similar to previous tests but also allow for continuity of longitudinal data collection and wide dissemination among research collaborators. A Crosswalk Study was conducted in early 2014 to assess the correlation between each set of new and previous tests. Tests with good correlation were equated using equipercentile equating. The resulting conversion tables allow scores on the new tests to be converted to equivalent scores on the previous tests. All pairs of tests had good correlation (ρ=0.68 to 0.78). Learning effects were detected for Logical Memory only. Confidence intervals were narrow at each point estimate, and prediction accuracy was high. The recommended new tests are well correlated with the previous tests. The equipercentile equating method produced conversion tables that provide a useful reference for clinicians and researchers.


Improved mammography with a reduced radiation dose.
A 2-cm-high focused air-interspaced grid with a 7:1 ratio and eight lamellae per inch was devised. The grid is arc-shaped and pivots on its focal point, the anode-cathode axis. Milliamperage was 35% less for breast phantom radiographs obtained with the new grid versus those obtained with a conventional 5:1 mammographic grid. In a comparison study of five mammograms, each evaluated for five parameters, the images obtained with the new grid were judged to be of equal or better quality than those obtained with a conventional grid in 17 of 20 comparisons.


[Simultaneous rapid determination of 12 microcystins and one nodularin in water by direct injection-ultra performance liquid chromatography-triple quadrupole mass spectrometry].
A rapid method was developed for the simultaneous determination of 12 microcystins (MCs) and one nodularin (NOD) in water by direct injection-ultra performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-MS/MS). The water samples were first diluted with equal volume of methanol, and then filtered through polyether sulfone (PES) syringe filter. The filtrates were directly injected into the UPLC system. The separation of the analytes was carried out on an ACQUITY UPLC BEH 300 C18 column (100 mm×2.1 mm, 1.7 μm) with gradient elution using mobile phases of acetonitrile containing 0.1% (v/v) formic acid and 0.2% (v/v) formic acid aqueous solution. The 12 microcystins and one nodularin were detected by positive electrospray ionization in the multiple reaction monitoring (MRM) mode, and quantified by standard solvent external standard method. The limits of detection were 0.03-0.1 μg/L and the limits of quantification were 0.1-0.3 μg/L. The recoveries were in the range of 79.5%-123% with the relative standard deviations ranging from 1.0% to 20% (n=6). The method is simple, sensitive and accurate, and has been successfully applied to the detection of the 13 kinds of algae toxins in water.


Learning in clinical nursing practice.
In this study, a qualitative interpretivist perspective was used to analyze the following research question: How does clinical nursing practice facilitate learning? Forty nurses working in acute care, long-term care, and home care were interviewed about their clinical practice experiences and the learning that resulted. Data were analyzed using a constant comparative method. Study results indicate that clinical nursing practice facilitates both personal and professional development of nurses by fostering an examination of the concepts of self-respect, hope, control, vulnerability, acceptance, loss, and persistence. Implications for continuing educators and staff development instructors are drawn.


Systematic studies on the chemical structure and umami enhancing activity of Maillard-modified guanosine 5'-monophosphates.
Recent investigations on taste active principles in nucleotide rich yeast extracts led to the discovery of (R)- and (S)-N(2)-(1-carboxyethyl)-guanosine 5'-monophosphate as previously not reported umami enhancing molecules formed upon the Maillard reaction of guanosine 5'-monophosphate (5'-GMP) with dihydroxyacetone and glyceraldehyde, respectively. In the present study, systematic Maillard-type model reactions were performed with 5'-GMP and a homologous series of monosaccharides exhibiting a C(3)- to C(6)-carbon skeleton as well as with the reducing disaccharide maltose in the presence of an amino acid. By preparative RP-HPLC, various (R)- and (S)-N(2)-(1-carboxyalkyl)-guanosine 5'-monophosphates and (R)- and (S)-N(2)-(1-alkylamino)carbonylalkyl)guanosine 5'-monophosphates were isolated and identified by means of LC-MS, LC-TOF-MS, and 1D/2D-NMR spectroscopy. Sensory evaluation of these Maillard products revealed β-values for umami enhancement between 0.06 and 7.0 and identified a strong influence of the stereochemistry as well as the chain length of the N(2)-substituent on the umami enhancing activity. For all of the compounds evaluated, the (S)-configured isomers showed higher taste impact, whereas the (R)-isomers showed only marginal β-values, thus underlining the stereospecifity of the umami taste receptor binding site.


Is There Any Association Between Chronic Periodontitis and Anxiety in Adults? A Systematic Review.
Periodontitis is a multifactorial disease triggered by dysbiotic biofilms, involving the host's immune response, systemic and behavioral factors, including psychosocial conditions. This systematic review aimed to investigate the possible association between periodontitis and anxiety in adults. Searches were performed in PubMed, Scopus, Web of Science, Lilacs, Cochrane, and OpenGrey databases, without language restrictions, considering studies in adults (P-Participants), with (E-Exposure) and without periodontitis (C- Comparison) in an outcome of association with anxiety (O-outcome). Methodological quality assessment was carried out using the Newcastle-Ottawa protocol for case-control and cross-sectional studies, followed by an analysis of the level of evidence using the GRADE tool. Metanalysis was not performed due to several differences in methods applied by authors in primary studies. Eleven observational studies were selected according to the inclusion criteria from the total of 6,380 studies retrieved from databases. Eight studies demonstrated higher anxiety levels in subjects with periodontitis, among which only one study presented a high risk of bias. The GRADE tool revealed a low level of evidence for the anxiety outcome measured by the State-Trait Anxiety Inventory (STAI), both for case-control and cross-sectional studies. However, since anxiety may affect the quality of life of many subjects, it reinforces the need for further studies that evaluate this association for more extended periods. Clinical Trial Registration:PROSPERO-CRD42020190445.


Enhancing the sensitivity of mid-IR quantum cascade laser-based cavity-enhanced absorption spectroscopy using RF current perturbation.
The sensitivity of mid-IR quantum cascade laser (QCL) off-axis cavity-enhanced absorption spectroscopy (CEAS), often limited by cavity mode structure and diffraction losses, was enhanced by applying a broadband RF noise to the laser current. A pump-probe measurement demonstrated that the addition of bandwidth-limited white noise effectively increased the laser linewidth, thereby reducing mode structure associated with CEAS. The broadband noise source offers a more sensitive, more robust alternative to applying single-frequency noise to the laser. Analysis of CEAS measurements of a CO(2) absorption feature at 1890 cm(-1) averaged over 100 ms yielded a minimum detectable absorption of 5.5×10(-3) Hz(-1/2) in the presence of broadband RF perturbation, nearly a tenfold improvement over the unperturbed regime. The short acquisition time makes this technique suitable for breath applications requiring breath-by-breath gas concentration information.


[Case of complete denture with improvement of facial aspect through the recovery of the mandibular position].
This is a clinical case report of esthetic and masticatory impairment due to a mandibular protrusion during occlusion. The patient was a complete edentulous subject who complained about esthetic and matiscatory disturbance. First of all, treatment dentures were done, in order to improve the mandible position and recover the facial aspect. Objective evaluations were carried out to compare the improvements achieved after the insertion of the new dentures. The treatment dentures were used to improve patient's mandible position as well as vertical dimension. Once the facial appearance was recovered as a result of an appropriate mandible position, a reduction of the angle formed between the FH plane and the A-B plane from 87 to 83 was recognized after the insertion of the new dentures through the analysis of lateral cephalograms. It indicates that the mandible's position changed to backward position resulting in an improvement of the facial aspect. In the present clinical case report, not only subjective evaluation for appropriate mandible position and vertical dimension, but also including objective evaluation methods allowed us to evaluate and compare objectively the patient's pre and post operative condition.


Association of impulsivity and polymorphic microRNA-641 target sites in the SNAP-25 gene.
Impulsivity is a personality trait of high impact and is connected with several types of maladaptive behavior and psychiatric diseases, such as attention deficit hyperactivity disorder, alcohol and drug abuse, as well as pathological gambling and mood disorders. Polymorphic variants of the SNAP-25 gene emerged as putative genetic components of impulsivity, as SNAP-25 protein plays an important role in the central nervous system, and its SNPs are associated with several psychiatric disorders. In this study we aimed to investigate if polymorphisms in the regulatory regions of the SNAP-25 gene are in association with normal variability of impulsivity. Genotypes and haplotypes of two polymorphisms in the promoter (rs6077690 and rs6039769) and two SNPs in the 3' UTR (rs3746544 and rs1051312) of the SNAP-25 gene were determined in a healthy Hungarian population (N = 901) using PCR-RFLP or real-time PCR in combination with sequence specific probes. Significant association was found between the T-T 3' UTR haplotype and impulsivity, whereas no association could be detected with genotypes or haplotypes of the promoter loci. According to sequence alignment, the polymorphisms in the 3' UTR of the gene alter the binding site of microRNA-641, which was analyzed by luciferase reporter system. It was observed that haplotypes altering one or two nucleotides in the binding site of the seed region of microRNA-641 significantly increased the amount of generated protein in vitro. These findings support the role of polymorphic SNAP-25 variants both at psychogenetic and molecular biological levels.


[Dynamics of mortality of able-bodied population in CIS countries].
Comparative analysis founded on WHO European database was carried out concerning mortality in progress as compared between Russia and CIS countries. It is testified that the Russian Federation is distinguished by excelling rate of reduction of able-bodied population of middle age group. It is suggested that in this process significant role is played by health care system defects along with life-style factors. Motivation for enhancing activity and attention of medical workers as related to this age group, especially males is needed.


Sterol composition of a delta 5,7-sterol-rich strain of Saccharomyces cerevisiae during batch growth.
Sterol composition was examined during batch growth on complex media containing ethanol, molasses or glucose as the carbon source. The molasses-grown cells exhibited a balanced sterol composition throughout growth, maintaining the proportion of ergosterol to 24:28-dehydroergosterol equal to 1.4. The negative effect of glucose on sterol synthesis manifested itself by decreasing the accumulation of 24:28-dehydroergosterol and total sterols but not of ergosterol. Using ethanol as the sole carbon source, a large amount of 24:28-dehydroergosterol accumulated, partly at the expense of other sterols. The gradual addition of nitrogen source during growth significantly decreased the accumulation of ergosterol, 24:28-dehydroergosterol and of total sterols. A general scheme of regulation of sterol synthesis in baker's yeast is presented.


Determination of caffeine and its metabolites in urine by high-performance liquid chromatography and capillary electrophoresis.
Caffeine (CA) and its 14 main metabolites were determined in urine by reversed-phase high-performance liquid chromatography (RP-HPLC) and capillary electrophoresis (CE). After addition of 1,3,9-trimethylxanthine, uracil and beta-hydroxyethyltheophylline as internal standards, samples were separated by RP-HPLC into three fractions; A, B and C. The fractions were concentrated by lyophilization and analysed quantitatively by CE. Fraction A contained 5-acetylamino-6-amino-3-methyluracil, 3-methyluric acid, 7-methyluric acid and 1-methyluric acid. Fraction B contained 7-methylxanthine, 3-methylxanthine, 3,7-dimethyluric acid, 1-methylxanthine, 1,3-dimethyluric acid and 3,7-dimethylxanthine, while fraction C contained 1,7-dimethyluric acid, 1,7-dimethylxanthine, 1,3-dimethylxanthine, 1,3,7-trimethyluric acid and caffeine itself. The detection limit for the various metabolites ranged from 2-5 mumol l-1. The within-run precision for the metabolites ranged between 3.6% and 15.2%. The combination of HPLC and CE techniques was found to be a practical and specific method for determination of CA and its metabolites in urine.


Effects of ultrasonically nebulized distilled water on respiratory resistance and breathing pattern in normals and asthmatics.
The purpose of this study was to analyze changes in respiratory resistance (Rrs) and breathing pattern in normal and asthmatic subjects after exposure to ultrasonically nebulized distilled water (UNDW). After measurement of baseline Rrs and breathing pattern, ten normals inhaled UNDW for 30, 60, 120, 240 and 480 s administered at 15 min intervals. After the 480 s exposure, subjects inhaled two puffs of metaproterenol. Rrs was measured immediately before and after each exposure to UNDW and after metaproterenol while breathing pattern was continuously monitored. Ten asthmatics were exposed to UNDW and metaproterenol in a similar time sequence, but the durations of exposure to UNDW were 15, 30, 60, 120 and 240 s. Mean (+/- SD) and frequency histograms of minute ventilation (VE), tidal volume (VT), frequency, inspiratory time, fractional inspiratory time, mean inspiratory flow (VT/TI) and end-expiratory level at baseline were compared to values after each exposure to UNDW and after metaproterenol. All subjects experienced laryngeal irritation or cough during the exposure. In normals, there were no changes in Rrs or any of the components of breathing pattern. In asthmatics, 15 and 30 s exposures produced no changes in Rrs or breathing pattern. With subsequent longer exposures, mean Rrs showed a stepwise significant increase, along with parallel increases of VT/TI and end-expiratory level. VE increased after 60 and 120 s exposure, but reached a plateau after 240 s exposure despite continued increase in Rrs.(ABSTRACT TRUNCATED AT 250 WORDS)


Neurotrophin-3 gradients established by lentiviral gene delivery promote short-distance axonal bridging beyond cellular grafts in the injured spinal cord.
Neurotrophic factor delivery to sites of spinal cord injury (SCI) promotes axon growth into but not beyond lesion sites. We tested the hypothesis that sustained growth factor gradients beyond regions of SCI will promote significant axonal bridging into and beyond lesions. Adult rats underwent C3 lesions to transect ascending dorsal column sensory axons, and autologous bone marrow stromal cells were grafted into the lesion to provide a cellular bridge for growth into the injured region. Concurrently, lentiviral vectors expressing neurotrophin-3 (NT-3) or green fluorescent protein (GFP) (controls) were injected into the host cord rostral to the lesion to promote axon extension beyond the graft/lesion. Four weeks later, NT-3 gradients beyond the lesion were detectable by ELISA in animals that received NT-3-expressing lentiviral vectors, with highest average NT-3 levels located near the rostral vector injection site. Significantly more ascending sensory axons extended into tissue rostral to the lesion site in animals injected with NT-3 vectors compared with GFP vectors, but only if the zone of NT-3 vector transduction extended continuously from the injection site to the graft; any "gap" in NT-3 expression from the graft to rostral tissue resulted in axon bridging failure. Despite axon bridging beyond the lesion, regenerating axons did not continue to grow over very long distances, even in the presence of a continuing growth factor gradient beyond the lesion. These findings indicate that a localized and continuous gradient of NT-3 can achieve axonal bridging beyond the glial scar, but growth for longer distances is not sustainable simply with a trophic stimulus.


HER-2 gene amplification, HER-2 and epidermal growth factor receptor mRNA and protein expression, and lapatinib efficacy in women with metastatic breast cancer.
Biomarkers from two randomized phase III trials were analyzed to optimize selection of patients for lapatinib therapy. In available breast cancer tissue from EGF30001 (paclitaxel +/- lapatinib in HER-2-negative/unknown metastatic breast cancer, n = 579) and EGF100151 (capecitabine +/- lapatinib in HER-2-positive metastatic breast cancer, n = 399), HER-2 gene amplification by fluorescence in situ hybridization (FISH), HER-2 mRNA by reverse transcription-PCR (RT-PCR), HER-2 protein expression by HercepTest immunohistochemistry (IHC), epidermal growth factor receptor (EGFR) mRNA level by RT-PCR, and EGFR protein by IHC were analyzed and compared with clinical outcome. HER-2 was determined by FISH in an academic reference/research laboratory and in a large, high-volume commercial reference laboratory. The HER-2 gene was amplified in 47% (344 of 733) and IHC was 3+ in 35% (279 of 798), with significant correlation (P < 0.01) between FISH and IHC. Positive EGFR immunostaining (IHC 1+, 2+, or 3+) in 28% (213 of 761) correlated with EGFR mRNA levels by RT-PCR (r = 0.59; P < 0.01). HER-2 gene amplification/overexpression was associated with improved clinical outcomes (progression-free survival; P < 0.001) in both trials. A significant improvement in outcome was seen in FISH-positive and IHC 0, 1+, or 2+ patients. HER-2 mRNA expression correlated with HER-2 FISH (r = 0.83) and IHC status (r = 0.72; n = 138). No correlation was found between EGFR expression (IHC or mRNA) and responsiveness to lapatinib regardless of HER-2 status. Although a significant correlation with lapatinib responsiveness was observed among "HER-2-negative" breast cancer patients in the large, high-volume commercial reference laboratory, this was not confirmed in the academic reference/research laboratory. Women with HER-2-positive metastatic breast cancer benefit from lapatinib, whereas women with HER-2-negative metastatic breast cancer derive no incremental benefit from lapatinib.


A Sectoral Analysis of Vessel Density Measurements in Perimetrically Intact Regions of Glaucomatous Eyes: An Optical Coherence Tomography Angiography Study.
To study sectoral vessel density (VD) and structural alterations in the peripapillary and parafoveal hemiretina corresponding to perimetrically intact regions of glaucomatous eyes with hemifield defects and to compare these with healthy eyes using optical coherence tomography. This cross-sectional study included 37 eyes with open-angle glaucoma having visual fields defects restricted to 1 hemifield and 45 age-matched controls. Peripapillary VD and retinal nerve fiber layer (RNFL) thickness were measured in 8 sectors. Parafoveal VD and ganglion cell complex thickness were measured in the superior and inferior hemispheres of the macula. These parameters were compared between the intact hemiretinae of glaucomatous eyes and corresponding sectors of healthy eyes using the t test. Within the perimetrically intact regions of glaucomatous eyes, the mean VD and RNFL thickness were significantly reduced in inferotemporal, superonasal, and nasal upper peripapillary sectors as compared with healthy eyes (P<0.008). The temporal upper sector had reduced mean VD (60.3% vs. 62.9%, P=0.04) despite similar mean RNFL thickness (P=0.18). The superotemporal sector had decreased mean RNFL thickness (121 vs. 138 µm, P=0.0001) despite similar VD (P=0.06). At the macula, ganglion cell complex thickness was reduced in the superior and inferior hemispheres, but mean VD was reduced in the superior parafoveal region only. Peripapillary and parafoveal vascular changes precede functional decline. The extent of VD reduction and RNFL thinning varies in different peripapillary sectors and longitudinal studies are required to better understand the temporal relationship of vascular and RNFL loss.


Evaluation of the new cesium-131 seed for use in low-energy x-ray brachytherapy.
Characterization measurements and calculations were performed on a new medical seed developed by IsoRay Inc. in Richland, Washington, that utilizes the short-lived isotope 131Cs. This model has recently received FDA 510(k) clearance. The objective of this work was to characterize the dosimetric properties of the new seed according to the AAPM Task Group 43 recommendations. Cesium-131 is a low-energy x-ray emitter, with the most prominent peaks in the 29 keV to 34 keV region. The intended application is brachytherapy for treating cancers in prostate, breast, head and neck, lung, and pancreas. The evaluations performed included air-kerma strength, radial dose function, anisotropy in phantom, half-life, energy spectra, and internal activity. The results indicate the CS-1 seeds have a dose-rate constant of 0.915 cGy hr(-1) U(-1) in water, dose penetration characteristics similar to 125I and 103Pd, anisotropy function values on the order of 0.71 at short distances and small angles, and an average anisotropy factor of 0.964. The overall dosimetric characteristics are similar to 125I and 103Pd seeds with the exception of half-life, which is 9.7 days, as compared to 17 days for 103Pd and 60 days for 125I. The shorter half-life may offer significant advantages in biological effectiveness.


Human papillomavirus and skin tags: is there any association?
Low-risk human papillomavirus (HPV) infections are related to the genesis of various benign lesions. In an isolated report available, HPVs have been implicated in the causation of skin tags too. The present study was designed to detect the existence of low-risk HPV types 6 and 11 in cutaneous soft fibromas (skin tag) in north Indians. A total of 37 cases of skin tags from various sites were analyzed. Highly sensitive and comprehensive polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) assays were done for the detection of low-risk HPV types 6 and 11. The results revealed the presence of HPV DNA 6/11 in 48.6% of the skin tags examined by PCR-RFLP. This result corroborates the hypothesis that HPV plays a part in the etiology of benign lesions like cutaneous soft fibromas. The identification of HPV 6/11 in these lesions, which are benign proliferations of the skin, further expands the spectrum of HPV-linked lesions.


Modifications to the Patient Rule-Induction Method that utilize non-additive combinations of genetic and environmental effects to define partitions that predict ischemic heart disease.
This article extends the Patient Rule-Induction Method (PRIM) for modeling cumulative incidence of disease developed by Dyson et al. (Genet Epidemiol 31:515-527) to include the simultaneous consideration of non-additive combinations of predictor variables, a significance test of each combination, an adjustment for multiple testing and a confidence interval for the estimate of the cumulative incidence of disease in each partition. We employ the partitioning algorithm component of the Combinatorial Partitioning Method to construct combinations of predictors, permutation testing to assess the significance of each combination, theoretical arguments for incorporating a multiple testing adjustment and bootstrap resampling to produce the confidence intervals. An illustration of this revised PRIM utilizing a sample of 2,258 European male participants from the Copenhagen City Heart Study is presented that assesses the utility of genetic variants in predicting the presence of ischemic heart disease beyond the established risk factors.


Tuning the exponential sensitivity of a bound-state-in-continuum optical sensor.
In this work, we investigate the evanescent field sensing mechanism provided by an all-dielectric metasurface supporting bound states in the continuum (BICs). The metasurface is based on a transparent photonic crystal with subwavelength thickness. The BIC electromagnetic field is localized along the direction normal to the photonic crystal nanoscale-thin slab (PhCS) because of a topology-induced confinement, exponentially decaying in the material to detect. On the other hand, it is totally delocalized in the PhCS plane, which favors versatile and multiplexing sensing schemes. Liquids with different refractive indices, ranging from 1.33 to 1.45, are infiltrated in a microfluidic chamber bonded to the sensing dielectric metasurface. We observe an experimental exponential sensitivity leading to differential values as large as 226 nm/RIU with excellent FOM. This behavior is explained in terms of the physical superposition of the field with the material under investigation and supported by a thorough numerical analysis. The mechanism is then translated to the case of molecular adsorption where a suitable theoretical engineering of the optical structure points out potential sensitivities as large as 4000 nm/RIU.


Mechanical response of packings of nonspherical particles: A case study of two-dimensional packings of circulo-lines.
We investigate the mechanical response of jammed packings of circulo-lines in two spatial dimensions, interacting via purely repulsive, linear spring forces, as a function of pressure P during athermal, quasistatic isotropic compression. The surface of a circulo-line is defined as the collection of points that is equidistant to a line; circulo-lines are composed of a rectangular central shaft with two semicircular end caps. Prior work has shown that the ensemble-averaged shear modulus for jammed disk packings scales as a power law, 〈G(P)〉∼P^{β}, with β∼0.5, over a wide range of pressure. For packings of circulo-lines, we also find robust power-law scaling of 〈G(P)〉 over the same range of pressure for aspect ratios R≳1.2. However, the power-law scaling exponent β∼0.8-0.9 is much larger than that for jammed disk packings. To understand the origin of this behavior, we decompose 〈G〉 into separate contributions from geometrical families, G_{f}, and from changes in the interparticle contact network, G_{r}, such that 〈G〉=〈G_{f}〉+〈G_{r}〉. We show that the shear modulus for low-pressure geometrical families for jammed packings of circulo-lines can both increase and decrease with pressure, whereas the shear modulus for low-pressure geometrical families for jammed disk packings only decreases with pressure. For this reason, the geometrical family contribution 〈G_{f}〉 is much larger for jammed packings of circulo-lines than for jammed disk packings at finite pressure, causing the increase in the power-law scaling exponent for 〈G(P)〉.


Electrogram prolongation and nifedipine-suppressible ventricular arrhythmias in mice following targeted disruption of KCNE1.
Mutations in KCNE1, the gene encoding the beta subunit of the slowly activating delayed rectifier potassium current (IKs) channel protein, may lead to the long QT syndrome (LQTS), a condition associated with enhanced arrhythmogenesis. Mice with homozygous deletion of the coding sequence of KCNE1 have inner ear defects strikingly similar to those seen in the corresponding human condition. The present study demonstrated and assessed the mechanism of ventricular arrhythmias in Langendorff-perfused whole heart preparations from homozygous KCNE1-/- mice compared to wild-type mice of the same age. The effects of programmed electrical stimulation with decremental pacing from the basal right ventricular epicardial surface upon electrogram waveforms recorded from the basal left ventricle were assessed and quantified using techniques of paced electrogram fractionation analysis for the first time in an experimental system. All KCNE1-/-(n = 10) but not wild-type (n = 14) mouse hearts empirically demonstrated marked pacing-induced ventricular arrhythmogenicity. This correlated with significant increases in electrogram dispersion, consistent with a wider spread in conduction velocities, in parallel with clinical findings from LQTS patients with potassium channel mutations. In contrast, introduction of 100 nM isoprenaline induced arrhythmogenicity in both KCNE1-/- (n = 7) and wild-type (n = 6) hearts during pacing. Furthermore, pretreatment with 1 muM nifedipine exerted a strong anti-arrhythmic effect in the KCNE1-/- hearts (n = 12) that persisted even in the presence of 100 nM isoprenaline (n = 6). Our findings associate KCNE1-/- with an arrhythmogenic phenotype that shows an increased dispersion of conduction velocities, and whose initiation is prevented by nifedipine, a finding that in turn may have therapeutic applications in conditions such as LQTS.


Enhanced diabetic wound healing using platelet-derived extracellular vesicles and reduced graphene oxide in polymer-coordinated hydrogels.
Impaired wound healing is a significant complication of diabetes. Platelet-derived extracellular vesicles (pEVs), rich in growth factors and cytokines, show promise as a powerful biotherapy to modulate cellular proliferation, angiogenesis, immunomodulation, and inflammation. For practical home-based wound therapy, however, pEVs should be incorporated into wound bandages with careful attention to delivery strategies. In this work, a gelatin-alginate hydrogel (GelAlg) loaded with reduced graphene oxide (rGO) was fabricated, and its potential as a diabetic wound dressing was investigated. The GelAlg@rGO-pEV gel exhibited excellent mechanical stability and biocompatibility in vitro, with promising macrophage polarization and reactive oxygen species (ROS)-scavenging capability. In vitro cell migration experiments were complemented by in vivo investigations using a streptozotocin-induced diabetic rat wound model. When exposed to near-infrared light at 2 W cm- 2, the GelAlg@rGO-pEV hydrogel effectively decreased the expression of inflammatory biomarkers, regulated immune response, promoted angiogenesis, and enhanced diabetic wound healing. Interestingly, the GelAlg@rGO-pEV hydrogel also increased the expression of heat shock proteins involved in cellular protective pathways. These findings suggest that the engineered GelAlg@rGO-pEV hydrogel has the potential to serve as a wound dressing that can modulate immune responses, inflammation, angiogenesis, and follicle regeneration in diabetic wounds, potentially leading to accelerated healing of chronic wounds.


The usability of WeChat as a mobile and interactive medium in student-centered medical teaching.
Biochemistry and cellular biology courses for medical students at Tongji University include the assessment that provides students with feedback to enhance their learning, which is a type of formative assessment. However, frequent instant feedback and guidance for students is often absent or inconsistently included in the teaching process. WeChat, the most popular Chinese social media, was introduced in biochemistry and cellular biology course. A WeChat official account (OA) was set up as an instant interactive platform. Over a period of two semesters, OA sent 73 push notifications. The components included course notices, preclass thought questions, after-class study materials, answer questions and feedback, simulation exercises, teacher-student interaction, and research progress relevant to the course. WeChat OA served as an active-learning teaching tool, provided more frequent feedback and guidance to students, and facilitated better student-centered communication in the teaching process. Using the WeChat OA in medical teaching emphasized interactive, interoperable, effective, engaging, adaptable, and more participatory teaching styles. As a new platform, WeChat OA was free, Internet-reliant, and easily managed. Using this new medium as a communication tool accelerated further advancement of instant feedback and improvement in teaching activities. Notifications and interactive feedback via the mobile social medium WeChat OA anytime and anywhere facilitated a student-centered teaching mode. Use of WeChat OA significantly increased the proportion of students interactively participating and resulted in a high degree of student satisfaction. © 2017 by The International Union of Biochemistry and Molecular Biology, 45(5):421-425, 2017.


Superiority of Second Stage Ossiculoplasty.
To compare hearing results of single stage ossiculoplasty with second stage ossiculoplasty, and to provide a proper indication of operation options (single stage or second stage ossiculoplasty) according to patients' clinical conditions. Retrospective case review study. Tertiary referral center. A total of 175 patients with chronic otitis media. Tympanoplasty or mastoidectomy with tympanoplasty (M&T) as well as ossiculoplasty of either single or second stage between January 2009 and March 2016 by one surgeon (S.N.P.). Hearing outcomes evaluated by mean postoperative air-bone gap (ABG) and success rates in various subgroups of middle ear surgeries and different types of ossiculoplasty were analyzed. Among 175 patients, 120 underwent single stage ossiculoplasty, and 55 underwent second stage ossiculoplasty. The benefit of second stage ossiculoplasty in hearing results both at 3 months visit (p < 0.001) and 2 years visit (p = 0.003) were observed. The superiority of second stage ossiculoplasty was found in 20 dBHL cut off value with a statistical significance. The subgroups of canal wall down mastoidectomy and the first, non-revision middle ear surgery showed statistically significant benefits with second stage ossiculoplasty. Second stage ossiculoplasty has overall advantage in hearing results compared with single stage ossiculoplasty and could be more advantageous to the patient who needs canal wall down mastoidectomy or whose operation is the first middle ear surgery.


Absorption spectra of chlorophyll a and b in Lhcb protein environment.
The spectral forms of the two chlorophyll species in higher plant Photosystem II antenna proteins have been experimentally determined within their protein environment. Recombinant CP29 and LHC II antenna proteins missing individual chromophores were obtained by over-expression in bacteria without any changing of the primary protein sequence and in vitro reconstitution. Difference absorption spectroscopy with respect to the corresponding proteins binding the complete pigment complement yielded the spectral shape and extinction of single chlorophyll a and b. A functional relation of their absorption was given by Gaussian subband decomposition covering the entire Q(x) and Q(y) optical region together with the absolute value of the molar extinction coefficient. With respect to analogous determinations reported in the literature for organic solvents, this information is valuable for further understanding the in-protein chlorophyll excited states and excited state dynamics: in particular, for the calculation of Förster transfer rates by means of chlorophyll-chlorophyll overlap integral employing the Stepanov relation for emission and single chromophore transition energies according to the results of mutational analysis of chlorophyll binding sites [Bassi et al. (1999) Proc Natl Acad Sci USA 96: 10056-10061; Remelli et al. (1999) J Biol Chem 274: 33510-33521].


Independent usefulness of flow phase 99mTc-red blood cell scintigraphy in predicting the results of angiography in acute gastrointestinal bleeding.
In acute gastrointestinal bleeding, despite positive dynamic phase 99mTc-red blood cell scintigraphy, invasive catheter angiography (CA) is frequently negative. In this study, we investigated the value of flow phase scintigraphy in predicting extravasation on CA. Institutional review board approval with a waiver of informed consent was obtained for this retrospective study. A total of 173 scintigraphy procedures performed in 145 patients with GIB between January 2013 and August 2014 were analysed. Scintigraphy had two phases: flow (1 image/s for 1 min) followed by dynamic (1 image/30 s for 1 h). Patients who underwent CA within 24 hours of positive scintigraphy were assessed. Each scintigraphy phase was randomly and independently reviewed by two nuclear medicine physicians blinded to the outcomes of the other phase and of CA. A total of 42 patients (29%) had positive scintigraphy. Of these patients, 29 underwent CA, and extravasation was seen in 6 (21%). In all, dynamic phase scintigraphy was positive. 13 of the 29 patients also had positive flow phase scintigraphy. The sensitivity, specificity, positive-predictive value, and negative-predictive value of flow phase scintigraphy for extravasation on CA were 100, 70, 46, and 100%, respectively. Specificity and positive predictive value were higher when CA was performed within 4 hours of positive flow phase scintigraphy. Negative flow phase scintigraphy can identify patients who will not benefit from CA despite positive dynamic phase scintigraphy. The likelihood of extravasation on CA is higher when performed soon after positive flow phase scintigraphy. Negative flow phase scintigraphy identifies patients who will not benefit from invasive catheter angiography despite positive results on subsequent dynamic phase scintigraphy. Increasing the delay between positive red blood cell scintigraphy and catheter angiography progressively reduces the likelihood of identifying extravasation, which is required to target embolization.


Mesenchymal stem cells and conditioned medium avert enteric neuropathy and colon dysfunction in guinea pig TNBS-induced colitis.
Damage to the enteric nervous system (ENS) associated with intestinal inflammation may underlie persistent alterations to gut functions, suggesting that enteric neurons are viable targets for novel therapies. Mesenchymal stem cells (MSCs) offer therapeutic benefits for attenuation of neurodegenerative diseases by homing to areas of inflammation and exhibiting neuroprotective, anti-inflammatory, and immunomodulatory properties. In culture, MSCs release soluble bioactive factors promoting neuronal survival and suppressing inflammation suggesting that MSC-conditioned medium (CM) provides essential factors to repair damaged tissues. We investigated whether MSC and CM treatments administered by enema attenuate 2,4,6-trinitrobenzene-sulfonic acid (TNBS)-induced enteric neuropathy and motility dysfunction in the guinea pig colon. Guinea pigs were randomly assigned to experimental groups and received a single application of TNBS (30 mg/kg) followed by 1 × 10(6) human bone marrow-derived MSCs, 300 μl CM, or 300 μl unconditioned medium 3 h later. After 7 days, the effect of these treatments on enteric neurons was assessed by histological, immunohistochemical, and motility analyses. MSC and CM treatments prevented inflammation-associated weight loss and gross morphological damage in the colon; decreased the quantity of immune infiltrate in the colonic wall (P < 0.01) and at the level of the myenteric ganglia (P < 0.001); prevented loss of myenteric neurons (P < 0.05) and damage to nerve processes, changes in ChAT, and nNOS immunoreactivity (P < 0.05); and alleviated inflammation-induced colonic dysmotility (contraction speed; P < 0.001, contractions/min; P < 0.05). These results provide strong evidence that both MSC and CM treatments can effectively prevent damage to the ENS and alleviate gut dysfunction caused by TNBS-induced colitis.


The prognostic significance of circulating serum amyloid A and CXC chemokine ligand 4 in osteosarcoma.
Osteosarcoma (OS) is the most common pediatric bone cancer. Despite advances in treatment regimens, the survival rate remains 60-70%. There is an urgent need to identify prognostic biomarkers, so that targeted therapies can be developed to improve the outcome. Our laboratory has previously identified that circulating serum amyloid A (SAA) and CXC chemokine ligand 4 (CXCL4) are upregulated in patients with OS. In this study, we tested if they could be used as prognostic biomarkers. We used enzyme-linked immunosorbent assays to measure their concentrations in serum samples (n = 233) and immunohistochemistry to examine their expressions in primary tumors (n = 37). Prognostic significance of the serum concentrations and tumor expressions of the biomarkers was then evaluated. Patients with "high SAA" and "low CXCL4" circulating levels at diagnosis significantly correlated with a worse outcome (HR = 1.68, P = 0.014), which was independent of the metastatic status. These patients also exhibited a significantly higher rate of poor histologic response to chemotherapy. Furthermore, low tumor expression of CXCL4 correlated with poor survival (HR = 3.57, P = 0.005). Our results demonstrate that circulating SAA and CXCL4 may serve as prognostic biomarkers in OS. Targeting CXCL4 has been reported, suggesting that it may be exploited as a therapeutic target in OS.


Cu-Catalyzed Switchable Asymmetric Defluoroalkylation and [3 + 2] Cycloaddition of Trifluoropropene.
Chiral fluorinated amino esters and pyrrolidines are privileged scaffolds in synthetic chemistry and exhibit unique biological properties. We report the facile preparation of these compounds through copper-catalyzed switchable defluoroalkylation and [3 + 2] cycloaddition of trifluoropropene in an asymmetric fashion. The choice of solvent and chiral ligand was crucial for the efficient transformation and exquisite chemoselectivity pattern from identical starting materials that rapidly and reliably incorporate gem-difluoroalkene and trifluoromethyl (CF3) motifs to generate a diverse range of enantioenriched fluorinated building blocks in good to excellent yields with high asymmetric induction.


Microbial inactivation of Pseudomonas putida and Pichia pastoris using gene silencing.
Antisense deoxyoligonucleotide (ASO) gene silencing was investigated as a potential disinfection tool for industrial and drinking water treatment application. ASOs bind with their reverse complementary mRNA transcripts thereby blocking protein translation. While ASO silencing has mainly been studied in medicine, it may be useful for modulating gene expression and inactivating microorganisms in environmental applications. In this proof of concept work, gene targets were sh ble (zeocin resistance) and todE (catechol-2,3-dioxygenase) in Pichia pastoris and npt (kanamycin resistance) in Pseudomonas putida. A maximum 0.5-fold decrease in P. pastoris cell numbers was obtained following a 120 min incubation with single-stranded DNA (ssDNA) concentrations ranging from 0.2 to 200 nM as compared to the no ssDNA control. In P. putida, a maximum 5.2-fold decrease was obtained after 90 min with 400 nM ssDNA. While the silencing efficiencies varied for the 25 targets tested, these results suggest that protein activity as well as microbial growth can be altered using ASO gene silencing-based tools. If successful, this technology has the potential to eliminate some of the environmental and health issues associated with the use of strong chemical biocides. However, prior to its dissemination, more research is needed to increase silencing efficiency and develop effective delivery methods.


New analogues of acyclovir--synthesis and biological activity.
New acyclovir esters with peptidomimetics were synthesized and evaluated in vitro for their antiviral activity against the replication of Herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2). The influence of peptidomimetics containing oxazole and thiazolyl-thiazole moieties on the antiviral activity is also reported. The esters were synthesized using the coupling reagents N-ethyl-N'-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC) and N,N-dimethyl-4-aminopyridine (DMAP) as a catalyst.


Nuclear localization mechanism of Hsp105beta and its possible function in mammalian cells.
Hsp105alpha and Hsp105beta are mammalian stress proteins of the Hsp105/110 family. We have shown that Hsp105beta localizes to the nucleus, whereas Hsp105alpha localizes to the cytoplasm of mammalian cells. Hsp105alpha localizes in the cytoplasm, as the nuclear export signal (NES) activity rather than nuclear localization signal (NLS) activity dominates in Hsp105alpha, due to suppression of the NLS activity. In this study, we determined the mechanisms behind the nuclear localization of Hsp105beta, and revealed that the NES was suppressed by the N-terminal (amino acids 3-10) or C-terminal (amino acids 699-756) region of Hsp105beta, and the NLS activity rather than NES activity seemed to dominate in Hsp105beta. Furthermore, as Hsp105beta which localizes in the nucleus, functioned as an inducer of Hsp70 in mammalian cells, Hsp105 family proteins may play an important role in the protection of cells against deleterious stressor together with Hsp70.


Serum cytokeratin-18 fragment levels as noninvasive marker of nonalcoholic steatohepatitis in the chilean population.
Nonalcoholic steatohepatitis (NASH) is the most aggressive form of nonalcoholic fatty liver disease (NAFLD) and involves the risk of progression to more advanced stages of liver disease. Non-invasive methods are needed to identify patients with NASH. To evaluate the diagnostic performance of the determination of serum levels of cytokeratin-18 (CK-18) as a non-invasive marker of NASH in the Chilean population. Serum CK-18 levels were determined in a group of 41 patients with biopsy-proven NAFLD. NASH diagnosis was based on Brunt's criteria (histological parameters and ballooning), and the NAFLD activity score (NAS) and the presence of fibrosis were determined. The correlation between the NAFLD activity score (NAS) and CK-18 was evaluated with Spearman's rank correlation coefficient. A ROC curve was produced to assess the diagnostic value of CK-18 for NASH. The NAFLD fibrosis score (NFS) (to predict fibrosis and NASH) was compared to CK-18 with simple linear regression. Data were expressed in median [25th-75th percentile] and evaluated with the Wilcoxon rank test. The mean age of the study group (23% male) was 50.4±11.1 years. 34.2% were diagnosed with NASH (NAS≥5). CK-18 levels were significantly higher in patients with NASH versus those without NASH (183.6 IU/l [97.4 to 734.4] vs. 117.2 IU/l [83.8 to 954.8], p= 0.016). CK-18 levels were a good predictor of NASH on biopsy with an area under the curve (AUC) of 0.732 (95% CI, 0.572 to 0.897). A CK-18 cut-off of 130.5 IU/l had a sensitivity of 92.9%, specificity of 63%, positive predictive value of 56.5% and negative predictive value of 94.4%, and was able to correctly classify 73.2% of patients with NASH. NFS identified advanced liver fibrosis (AUC 0.739, 95% CI, 0.56-0.91), but was of limited value to identify NASH (AUC 0.413, 95% CI, 0.21-0.61). CK-18 is a good non-invasive marker for NASH. Although NFS was found to be an accurate marker of advanced liver fibrosis, it was not of value to identify NASH. In patients with NAFLD, CK-18 and NFS could be useful in predicting NASH and liver fibrosis, respectively.


Development of polymerase chain reaction-based diagnostic tests for detection of Malsoor virus & adenovirus isolated from Rousettus species of bats in Maharashtra, India.
Bats are recognized as important reservoirs for emerging infectious disease and some unknown viral diseases. Two novel viruses, Malsoor virus (family Bunyaviridae, genus, Phlebovirus) and a novel adenovirus (AdV) (family, Adenoviridae genus, Mastadenovirus), were identified from Rousettus bats in the Maharashtra State of India. This study was done to develop and optimize real time reverse transcription - polymerase chain reaction (RT-PCR) assays for Malsoor virus and real time and nested PCR for adenovirus from Rousettus bats. For rapid and accurate screening of Malsoor virus and adenovirus a nested polymerase chain reaction and TaqMan-based real-time PCR were developed. Highly conserved region of nucleoprotein gene of phleboviruses and polymerase gene sequence from the Indian bat AdV isolate polyprotein gene were selected respectively for diagnostic assay development of Malsoor virus and AdV. Sensitivity and specificity of assays were calculated and optimized assays were used to screen bat samples. Molecular diagnostic assays were developed for screening of Malsoor virus and AdV and those were found to be specific. Based on the experiments performed with different parameters, nested PCR was found to be more sensitive than real-time PCR; however, for rapid screening, real-time PCR can be used and further nested PCR can be used for final confirmation or in those laboratories where real-time facility/expertise is not existing. This study reports the development and optimization of nested RT-PCR and a TaqMan-based real-time PCR for Malsoor virus and AdV. The diagnostic assays can be used for rapid detection of these novel viruses to understand their prevalence among bat population.


Interaction and transport of poly(L-lysine) dendrigrafts through liposomal and cellular membranes: the role of generation and surface functionalization.
Two generations of poly(l-lysine) dendrigrafts (DGLs) were studied with regard to their ability to interact with and translocate through liposomal and cellular membranes. Partial guanidinylation of the surface amino groups of the starting dendrigrafts afforded the guanidinylated derivatives whose membrane translocation properties were also assessed. Mixed liposomes, consisting of dihexadecyl phosphate, phosphatidylcholine, and cholesterol, were employed as model membranes, while A549 human lung carcinoma cells were used for cellular uptake studies. At high surface group/liposomal phosphate molar ratios and depending on the structure of the DGL, the interaction led to aggregation. Dendrigraft liposomal internalization was achieved, however, at low molar ratios. Thus translocation of the second generation dendrigrafts was rather limited at 25 degrees C, which, however, was enhanced when the bilayer was in the liquid-crystalline phase. In contrast, third-generation counterparts exhibited minor translocational ability. Furthermore, the introduction of a guanidinium group to dendrigrafts was found to enhance their transport through liposomal membranes. On the other hand, cellular uptake by A549 cells was monitored up to 3 h incubation time via fluorescence registration employing fluorescein-labeled dendrigrafts. The efficiency of dendrigraft internalization was enhanced by the presence of the guanidinium groups, while DGLs were preferentially localized in the nucleus and nuclear membrane, as revealed by fluorescence microscopy.


Specificity and regulation of renal sulfate transporters.
Sulfate is essential for normal cellular function. The kidney plays a major role in sulfate homeostasis. Sulfate is freely filtered and then undergoes net reabsorption in the proximal tubule. The apical membrane Na(+)/sulfate cotransporter NaS1 (SLC13A1) has a major role in mediating proximal tubule sulfate reabsorption, as demonstrated by the findings of hyposulfatemia and hypersulfaturia in Nas1-null mice. The anion exchanger SAT1 (SLC26A1), the founding member of the SLC26 sulfate transporter family, mediates sulfate exit across the basolateral membrane to complete the process of transtubular sulfate reabsorption. Another member of this family, CFEX (SLC26A6), is present at the apical membrane of proximal tubular cells. It also can transport sulfate by anion exchange, which probably mediates backflux of sulfate into the lumen. Knockout mouse studies have demonstrated a major role of CFEX as an apical membrane Cl(-)/oxalate exchanger that contributes to NaCl reabsorption in the proximal tubule. Several additional SLC26 family members mediate sulfate transport and show some level of renal expression (e.g., SLC26A2, SLC26A7, SLC26A11). Their roles in mediating renal tubular sulfate transport are presently unknown. This paper reviews current data available on the function and regulation of three sulfate transporters (NaS1, SAT1, and CFEX) and their physiological roles in the kidney.


Intelligence benefit of the 3-methoxytyramine to tyramine ratio in equine urine.
Equine urine analysis has evolved over time to detect thousands of urinary compounds for doping control in the horse racing industry. The longitudinal assessment of 3-methoxytyramine to tyramine ratio (3-MT/T) values in equine urine by GC-MS profiling was investigated to support the Racing NSW Equine Biological Passport (EBP) for detection of dopaminergic manipulation in racehorses. This involved comparison of routine urine samples to administration studies of Sinemet, a common Parkinson's disease medication containing levodopa. Using an endogenous reference compound (ERC) in a urinary ratio enabled greater confidence to provide intelligence of pharmaceutical manipulation as distinct from physiological variation. Population reference limits (PRLs) of 776 ng/ml for urinary 3-MT and 5.3 for 3-MT/T, together with the use of individual reference limits (IRLs), are proposed.


Tumor-Derived Exosomes Induce the Formation of Neutrophil Extracellular Traps: Implications For The Establishment of Cancer-Associated Thrombosis.
Cancer patients are at an increased risk of developing thromboembolic complications. Several mechanisms have been proposed to explain cancer-associated thrombosis including the release of tumor-derived extracellular vesicles and the activation of host vascular cells. It was proposed that neutrophil extracellular traps (NETs) contribute to the prothrombotic phenotype in cancer. In this study, we evaluated the possible cooperation between tumor-derived exosomes and NETs in cancer-associated thrombosis. Female BALB/c mice were orthotopically injected with 4T1 breast cancer cells. The tumor-bearing animals exhibited increased levels of plasma DNA and myeloperoxidase in addition to significantly increased numbers of circulating neutrophils. Mice were subjected to either Rose Bengal/laser-induced venous thrombosis or ferric chloride-induced arterial thrombosis models. The tumor-bearing mice exhibited accelerated thrombus formation in both models compared to tumor-free animals. Treatment with recombinant human DNase 1 reversed the prothrombotic phenotype of tumor-bearing mice in both models. Remarkably, 4T1-derived exosomes induced NET formation in neutrophils from mice treated with granulocyte colony-stimulating factor (G-CSF). In addition, tumor-derived exosomes interacted with NETs under static conditions. Accordingly, the intravenous administration of 4T1-derived exosomes into G-CSF-treated mice significantly accelerated venous thrombosis in vivo. Taken together, our observations suggest that tumor-derived exosomes and neutrophils may act cooperatively in the establishment of cancer-associated thrombosis.


Early childhood gut microbiomes show strong geographic differences among subjects at high risk for type 1 diabetes.
Gut microbiome dysbiosis is associated with numerous diseases, including type 1 diabetes. This pilot study determines how geographical location affects the microbiome of infants at high risk for type 1 diabetes in a population of homogenous HLA class II genotypes. High-throughput 16S rRNA sequencing was performed on stool samples collected from 90 high-risk, nonautoimmune infants participating in The Environmental Determinants of Diabetes in the Young (TEDDY) study in the U.S., Germany, Sweden, and Finland. Study site-specific patterns of gut colonization share characteristics across continents. Finland and Colorado have a significantly lower bacterial diversity, while Sweden and Washington state are dominated by Bifidobacterium in early life. Bacterial community diversity over time is significantly different by geographical location. The microbiome of high-risk infants is associated with geographical location. Future studies aiming to identify the microbiome disease phenotype need to carefully consider the geographical origin of subjects.


3D printing: Principles and pharmaceutical applications of selective laser sintering.
Pharmaceutical three-dimensional (3D) printing is a modern fabrication process with the potential to create bespoke drug products of virtually any shape and size from a computer-aided design model. Selective laser sintering (SLS) 3D printing combines the benefits of high printing precision and capability, enabling the manufacture of medicines with unique engineering and functional properties. This article reviews the current state-of-the-art in SLS 3D printing, including the main principles underpinning this technology, and highlights the diverse selection of materials and essential parameters that influence printing. The technical challenges and processing conditions are also considered in the context of their effects on the printed product. Finally, the pharmaceutical applications of SLS 3D printing are covered, providing an emphasis on the advantages the technology offers to drug product manufacturing and personalised medicine.


Janus Magnetic Nanoplatform for Magnetically Targeted and Protein/Hyperthermia Combination Therapies of Breast Cancer.
Protein therapeutics have been considered a promising strategy for cancer treatment due to their highly specific bioactivity and few side effects. Unfortunately, the low physiological stability and poor membrane permeability of most protein drugs greatly limit their clinical application. Furthermore, single-modality protein therapeutics show insufficient efficacy. To address these issues, Janus magnetic mesoporous silica nanoparticles (Janus MSNNPs) were developed to preload ribonuclease A (RNaseA) to simultaneously realize the magnetically enhanced delivery of protein drugs and magnetic hyperthermia-enhanced protein therapy. Janus MSNNPs showed a high RNaseA loading ability and pH-responsive drug release behavior. Furthermore, an external magnetic field could remarkably enhance the therapeutic effect of RNaseA-loaded Janus MSNNPs due to the improved intracellular internalization of RNaseA. Importantly, Janus MSNNPs possessed an outstanding magnetic hyperthermia conversion efficiency, which could generate hyperthermia under an alternating magnetic field, effectively supplementing protein therapy by a combined effect. In vitro and in vivo experiments confirmed the high anticancer outcome and low side effects of this intriguing strategy for breast cancer based on Janus MSNNPs. Hence, Janus MSNNPs might be an effective and safe nanoplatform for magnetically combined protein therapy.


Is there a benefit to drains with a Kocher-Langenbeck approach? A prospective randomized pilot study.
Closed suction drainage is a routine part of wound management for patients undergoing surgical treatment of acetabulum fractures. This pilot study seeks to determine if there is a difference in wound healing for a Kocher-Langenbeck approach with and without the use of drains. We conducted a prospective, randomized study including 39 patients with acetabulum fractures treated through a Kocher-Langenbeck approach. During wound closure, patients were randomized into two groups: 20 patients (group I) received drains and 19 (group II) were closed without drains. All were followed up for drain output, quality and quantity of drainage, signs of infection, and duration of drainage. Patients were then evaluated at 2 weeks and 8 weeks for wound healing and any signs of infection. By the 8-week follow-up, all wounds healed without any signs of infection. There was no difference in the average number of days of drainage between groups: 7.45 days and 7.95 days for group I and group II, respectively (p = 0.37). There were two wound complications (5.13%), with one in each group. Both complications consisted of cellulitis without signs of deep infection and had complete resolution with intravenous antibiotics. A post hoc power analysis determined that a test population of 1,264 patients would be needed to show a reduction in wound drainage time by 1 day. With the numbers available in this pilot study, we showed no benefit to the use of drains for acetabular surgery performed through a Kocher-Langenbeck approach.


A novel intra-operative, high-resolution atrial mapping approach.
A new technique is demonstrated for extensive high-resolution intra-operative atrial mapping that will facilitate the localization of atrial fibrillation (AF) sources and identification of the substrate perpetuating AF. Prior to the start of extra-corporal circulation, a 8 × 24-electrode array (2-mm inter-electrode distance) is placed subsequently on all the right and left epicardial atrial sites, including Bachmann's bundle, for recording of unipolar electrograms during sinus rhythm and (induced) AF. AF is induced by high-frequency pacing at the right atrial free wall. A pacemaker wire stitched to the right atrium serves as a reference signal. The indifferent pole is connected to a steal wire fixed to subcutaneous tissue. Electrograms are recorded by a computerized mapping system and, after amplification (gain 1000), filtering (bandwidth 0.5-400 Hz), sampling (1 kHz) and analogue to digital conversion (16 bits), automatically stored on hard disk. During the mapping procedure, real-time visualization secures electrogram quality. Analysis will be performed offline. This technique was performed in 168 patients of 18 years and older, with coronary and/or structural heart disease, with or without AF, electively scheduled for cardiac surgery and a ventricular ejection fraction above 40 %. The mean duration of the entire mapping procedure including preparation time was 9 ± 2 min. Complications related to the mapping procedure during or after cardiac surgery were not observed. We introduce the first epicardial atrial mapping approach with a high resolution of ≥1728 recording sites which can be performed in a procedure time of only 9±2 mins. This mapping technique can potentially identify areas responsible for initiation and persistence of AF and hopefully can individualize both diagnosis and therapy of AF.


Independent Supported Housing for Non-homeless People With Serious Mental Illness: A Pragmatic Randomized Controlled Trial.
Independent supported housing (ISH)-services to support independent housing are recommended by current guidelines. However, there is little evidence of ISH models for non-homeless people with severe mental illness (SMI). The aim of this study was to examine the effectiveness of ISH by comparing the clinical outcomes of a newly implemented ISH service with regular housing and support services. A total of 58 adults with a broad spectrum of mental disorders experiencing housing problems were randomly assigned to either the intervention group (IG) with the possibility to use the ISH service in Zurich providing targeted, individual and flexible support for housing problems or to the control group (CG) with regular housing and support services currently available (trial registration at ClinicalTrials.gov: NCT03815604). After 12 months, almost all participants of the IG were able to live independently and need for inpatient treatment could be significantly reduced. Service utilization varied between 2 and 79 h. In the CG, 70% of the participants wanted to join a waiting list for the ISH service. The results indicated that IHS was comparable to regular housing and support services in terms of social inclusion and other social and clinical outcomes such as quality of life, capabilities, needs, mental state and functioning (p's > 0.05). The costs of service utilization were on average 115 Swiss Francs (about 124 USD) per participant per month. ISH is an effective service in housing rehabilitation in terms of social and clinical outcomes and costs. ISH is strongly preferred by service users. In line with the UN Convention on the Rights of Persons with Disabilities, access to ISH services for non-homeless people with SMI should be improved. ClinicalTrials.gov, identifier: NCT03815604, December 04, 2019.


Heteroatom-Doped Carbon-Encapsulated FeP Nanostructure: A Multifunctional Electrocatalyst for Zinc-Air Battery and Water Electrolyzer.
The rational design and synthesis of efficient multifunctional electrocatalysts for renewable energy technologies is of significant interest. Herein, we demonstrate a novel approach for the synthesis of a nitrogen and phosphorus dual-doped mesoporous carbon-encapsulated iron phosphide (FeP@NPC) nanostructure and its multifunctional electrocatalytic activity toward an oxygen reduction reaction, oxygen evolution reaction, and hydrogen evolution reaction for zinc-air battery (ZAB) and alkaline water-splitting applications. FeP@NPC is obtained by the carbothermal reduction of the precursor complex [Fe(bpy)3](PF6)2 in the presence of melamine without any traditional phosphidating agent. The PF6- counteranion is used for the phosphidation of Fe. FeP@NPC obtained at 900 °C (FeP@NPC-900) exhibits excellent bifunctional oxygen electrocatalytic performance with a very low potential gap (ΔE = E1/2ORR - Ej10OER) of 670 mV. The ZAB device delivers a peak power density of 190.15 mW cm-2 (iR-corrected), specific capacity of 785 mA h gZn-1, and energy density of 706.5 Wh kgZn-1 at 50 mA cm-2. The ZAB exhibits excellent charge-discharge cycling stability for over 35 h with negligible voltaic efficiency loss (0.9%). Three CR2032 coin-cell-based ZABs made of an FeP@NPC-900 air cathode connected in series power 81 LEDs for 15 min. FeP@NPC-900 also has promising electrocatalytic activity toward water splitting in acidic as well as in alkaline pH. The benchmark current density of 10 mA cm-2 is achieved with a two-electrode alkaline water electrolyzer at a cell voltage of 1.65 V. ZAB-powered water electrolyzer is made by integrating two rechargeable ZABs connected in series with the two-electrode water electrolyzer. The ZAB powers the electrolyzer for 24 h without a significant loss in the open-circuit voltage. The catalyst retains its initial structural integrity even after continuous water electrolysis for 24 h.


Predictors of ambulance transport in patients with ST-elevation myocardial infarction.
Despite intense public awareness campaigns, many patients with ST-elevation myocardial infarction (STEMI) do not utilize Emergency Medical Services (EMS) transportation to the Emergency Department (ED). Predictors for mode of transport by EMS versus private vehicle in patients with an acute STEMI were investigated. Hypothesis It was hypothesized that patient characteristics, specifically older age, male sex, and a history of a prior cardiac intervention, would be associated with a higher likelihood of EMS utilization. A retrospective, observational cohort study was performed for all STEMI patients treated from April 1, 2007 through June 30, 2010 at an urban, academic ED with 24-hour cardiac catheterization available. Multivariable analyses with predetermined predictors (age, sex, prior cardiac intervention, weekend/evening arrival) were performed to investigate associations with mode of transport. Door-to-balloon (D2B) times were calculated. Of the 209 STEMI patients, 11 were excluded, leaving 198 for analysis. Median age was 60 years (IQR: 53-70), 138 (70%) arrived by private vehicle, and 60 (30%) by EMS. The primary analysis did not identify significant predictors for EMS, but a post-hoc model found that private insurance (OR 0.18; 95% CI, 0.07-0.45) was associated with fewer EMS transports. Although not statistically significant due to the great variability in time of arrival for STEMI patients transported by private vehicle, EMS transports had shorter D2B times. During business hours and weekend/evenings, EMS had D2B times of 50 (IQR: 42-61) and 58 minutes (IQR: 47-63), respectively, while private vehicle transports had median D2B times of 62 (IQR: 50-74) and 78 minutes (IQR: 66-106). Conclusion No associations between mode of transport and patient age, sex, weekend/evening presentation and history of a prior cardiac intervention were identified. Privately insured patients were less likely to use EMS when experiencing a STEMI. More effective ways are needed to educate the public on the importance of EMS activation when one is concerned for acute coronary syndrome.


Patterning nanoscale flow vortices in nanochannels with patterned substrates.
We report the generation of complex toroidal nanoscale vortices in nanochannel flows, through the employment of patterned surface patches with contrasting wettability characteristics. Through extensive molecular-dynamics simulations, we further delineate that complicated interactions between the wettability gradient-driven vortex structures and surface roughness-influenced flow features may give rise to interesting patterns in the flow field. We also establish the orientation specificities of these nanoscale vortices, and demonstrate that the surface roughness effects may potentially act to alter the flow rotationalities realized out of modulated surface patterning effects to a considerable extent, which may have interesting consequences toward designing nanoscale mixing elements.


Tobacco and alcohol in relation to male breast cancer: an analysis of the male breast cancer pooling project consortium.
The etiology of male breast cancer is poorly understood, partly due to its relative rarity. Although tobacco and alcohol exposures are known carcinogens, their association with male breast cancer risk remains ill-defined. The Male Breast Cancer Pooling Project consortium provided 2,378 cases and 51,959 controls for analysis from 10 case-control and 10 cohort studies. Individual participant data were harmonized and pooled. Unconditional logistic regression was used to estimate study design-specific (case-control/cohort) ORs and 95% confidence intervals (CI), which were then combined using fixed-effects meta-analysis. Cigarette smoking status, smoking pack-years, duration, intensity, and age at initiation were not associated with male breast cancer risk. Relations with cigar and pipe smoking, tobacco chewing, and snuff use were also null. Recent alcohol consumption and average grams of alcohol consumed per day were also not associated with risk; only one subanalysis of very high recent alcohol consumption (>60 g/day) was tentatively associated with male breast cancer (ORunexposed referent = 1.29; 95% CI, 0.97-1.71; OR>0-<7 g/day referent = 1.36; 95% CI, 1.04-1.77). Specific alcoholic beverage types were not associated with male breast cancer. Relations were not altered when stratified by age or body mass index. In this analysis of the Male Breast Cancer Pooling Project, we found little evidence that tobacco and alcohol exposures were associated with risk of male breast cancer. Tobacco and alcohol do not appear to be carcinogenic for male breast cancer. Future studies should aim to assess these exposures in relation to subtypes of male breast cancer.


Congenital temporomandibular joint ankylosis: clinical characterization and natural history of four unrelated affected individuals.
The objective of this study was to describe four unrelated patients affected by congenital temporomandibular joint ankylosis. Clinical delineation, natural history, and possible etiologies are discussed. Clinical aspects that are important for follow-up are outlined.


The Effect of Multiple Firings on the Shear Bond Strength of Porcelain to a New Millable Alloy and a Conventional Casting Alloy.
This study compared the effect of multiple firings on the shear bond strength (SBS) of porcelain to the new millable alloy (Ceramill Sintron) and a conventional casting alloy (4-all). Thirty-six cylindrical cores (6.8 × 9 mm) were made of millable and castable alloy through CAD/CAM and casting techniques, respectively (n = 18). In the center of each bar, a 4 × 4 × 2-mm shot of porcelain was fused. Having divided each group into 3 subgroups based on the number of firing cycles (3, 5, 7), the specimens were fixed in a universal testing machine and underwent a shear force test (1.5 mm/min crosshead speed) until fractured. Then the SBS values (MPa) were calculated, and the failure patterns were microscopically characterized as adhesive, cohesive, or mixed. Two-way ANOVA statistical test revealed that the number of porcelain firings had no significant effect on the SBS of any of the metal groups (p = 0.1); however, it was statistically higher in the millable group than the castable group (p < 0.05). Moreover, detecting the mixed failure pattern in all the specimens implied that the multiple firings had no significant effect on the failure pattern. The multiple porcelain firings had no significant effect on the SBS of porcelain to neither the millable nor castable alloys.


Novel amino acid-based surfactant for silicone emulsification and its application in hair care products: a promising alternative to quaternary ammonium cationic surfactants.
Quaternary ammonium cationic surfactants (ACSs) and N-[3-alkyl(12,14)oxy-2-hydroxypropyl]-l-arginine hydrochloride (N-AOHPA) were used to emulsify silicone. The potential of the resulting emulsions in hair conditioning products was investigated. The emulsions were prepared using a homogenizer and/or high-pressure homogenizer. ACSs and N-AOHPA were used as silicone emulsifiers. The stability of the emulsions was evaluated by measuring particle sizes, creaming fractions, polydispersity indexes and zeta potentials. Moreover, the N-AOHPA-stabilized emulsion was compared with the ACS-stabilized emulsion to evaluate the adsorption amount of silicone on healthy and bleached hair surfaces and the inhibitory effects on amino acid dissolution from bleached hair. The adsorption site of the N-AOHPA-stabilized emulsion was observed using a scanning electron microscope. For all surfactants, the silicone emulsions prepared using the high-pressure homogenizer were more stable than those prepared using the homogenizer. When N-AOHPA was used as the surfactant, the silicone emulsion was especially stable. Furthermore, the d50 value of the N-AOHPA-stabilized emulsion was smaller than that of the ACS-stabilized emulsion. The adsorption behaviour of the silicone droplets in the different emulsions varied depending on the nature of the surfactant and the preparation method. The amount of ACS-stabilized silicone adsorbed on healthy hair was higher than that adsorbed on bleached hair, especially when the emulsion was prepared using the homogenizer. In contrast, the amount of N-AOHPA-stabilized silicone adsorbed on bleached hair was high, and no differences were observed between the N-AOHPA-stabilized emulsions prepared using the homogenizer and high-pressure homogenizer. The emulsified droplets, especially the N-AOHPA-stabilized droplets prepared using the high-pressure homogenizer, prevented amino acid dissolution from bleached hair. It was concluded that the silicone droplet adsorption site affected the dissolution process because the small N-AOHPA-stabilized droplets adsorbed at cuticle edges. This study shows that N-AOHPA has good potential for use as an emulsifier in products used for improving the condition of damaged hair.


Modulation of PrfA activity in Listeria monocytogenes upon growth in different culture media.
PrfA is the major transcriptional activator of most virulence genes of Listeria monocytogenes. Its activity is modulated by a variety of culture conditions. Here, we studied the PrfA activity in the L. monocytogenes wild-type strain EGD and an isogenic prfA deletion mutant (EGDDeltaprfA) carrying multiple copies of the wild-type prfA or the mutant prfA* gene (strains EGDDeltaprfApPrfA and EGDDeltaprfApPrfA*) in response to growth in brain heart infusion (BHI), Luria-Bertani broth (LB) or a defined minimal medium (MM) supplemented with one of the three phosphotransferase system (PTS) carbohydrates, glucose, mannose and cellobiose, or the non-PTS carbon source glycerol. Low PrfA activity was observed in the wild-type strain in BHI and LB with all of these carbon sources, while PrfA activity was high in minimal medium in the presence of glycerol. EGDDeltaprfApPrfA*, expressing a large amount of PrfA* protein, showed high PrfA activity under all growth conditions. In contrast, strain EGDDeltaprfApPrfA, expressing an equally high amount of PrfA protein, showed high PrfA activity only when cultured in BHI, and not in LB or MM (in the presence of any of the carbon sources). A ptsH mutant (lacking a functional HPr) was able to grow in BHI but not in LB or MM, regardless of which of the four carbon sources was added, suggesting that in LB and MM the uptake of the used PTS carbohydrates and the catabolism of glycerol are fully dependent on the functional common PTS pathway. The BHI culture medium, in contrast, apparently contains carbon sources (supporting listerial growth) which are taken up and metabolized by L. monocytogenes independently of the common PTS pathway. The growth rates of L. monocytogenes were strongly reduced in the presence of large amounts of PrfA (or PrfA*) protein when growing in MM, but were less reduced in LB and only slightly reduced in BHI. The expression of the genes encoding the PTS permeases of L. monocytogenes was determined in the listerial strains under the applied growth conditions. The data obtained further support the hypothesis that PrfA activity correlates with the expression level and the phosphorylation state of specific PTS permeases.


Accommodating the cost of growth and swimming in fish-the applicability of exercise-induced growth to juvenile hapuku (Polyprion oxygeneios).
Induced-swimming can improve the growth and feed conversion efficiency of finfish aquaculture species, such as salmonids and Seriola sp., but some species, such as Atlantic cod, show no or a negative productivity response to exercise. As a possible explanation for these species-specific differences, a recent hypothesis proposed that the applicability of exercise training, as well as the exercise regime for optimal growth gain (ERopt growth), was dependent upon the size of available aerobic metabolic scope (AMS). This study aimed to test this hypothesis by measuring the growth and swimming metabolism of hapuku, Polyprion oxygeneios, to different exercise regimes and then reconciling the metabolic costs of swimming and specific dynamic action (SDA) against AMS. Two 8-week growth trials were conducted with ERs of 0.0, 0.25, 0.5, 0.75, 1, and 1.5 body lengths per second (BL s(-1)). Fish in the first trial showed a modest 4.8% increase in SGR over static controls in the region 0.5-0.75 BL s(-1) whereas the fish in trial 2 showed no significant effect of ER on growth performance. Reconciling the SDA of hapuku with the metabolic costs of swimming showed that hapuku AMS is sufficient to support growth and swimming at all ERs. The current study therefore suggests that exercise-induced growth is independent of AMS and is driven by other factors.


Neuro-ophthalmology of late-onset Tay-Sachs disease (LOTS).
Late-onset Tay-Sachs disease (LOTS) is an adult-onset, autosomal recessive, progressive variant of GM2 gangliosidosis, characterized by involvement of the cerebellum and anterior horn cells. To determine the range of visual and ocular motor abnormalities in LOTS, as a prelude to evaluating the effectiveness of novel therapies. Fourteen patients with biochemically confirmed LOTS (8 men; age range 24 to 53 years; disease duration 5 to 30 years) and 10 age-matched control subjects were studied. Snellen visual acuity, contrast sensitivity, color vision, stereopsis, and visual fields were measured, and optic fundi were photographed. Horizontal and vertical eye movements (search coil) were recorded, and saccades, pursuit, vestibulo-ocular reflex (VOR), vergence, and optokinetic (OK) responses were measured. All patients showed normal visual functions and optic fundi. The main eye movement abnormality concerned saccades, which were "multistep," consisting of a series of small saccades and larger movements that showed transient decelerations. Larger saccades ended earlier and more abruptly (greater peak deceleration) in LOTS patients than in control subjects; these changes can be attributed to premature termination of the saccadic pulse. Smooth-pursuit and slow-phase OK gains were reduced, but VOR, vergence, and gaze holding were normal. Patients with late-onset Tay-Sachs disease (LOTS) show characteristic abnormalities of saccades but normal afferent visual systems. Hypometria, transient decelerations, and premature termination of saccades suggest disruption of a "latch circuit" that normally inhibits pontine omnipause neurons, permitting burst neurons to discharge until the eye movement is completed. These measurable abnormalities of saccades provide a means to evaluate the effects of novel treatments for LOTS.


Computational Study of the Radical Mediated Mechanism of the Formation of C8, C5, and C4 Guanine:Lysine Adducts in the Presence of the Benzophenone Photosensitizer.
The oxidation of guanine by triplet benzophenone in the presence of lysine has been shown to produce mono- and dilysine-substituted spiroiminodihydantion products, 8-Lys-Sp and 5,8-diLys-Sp. The potential energy surfaces for C8, C5, and C4 nucleophilic addition have been mapped out using the B3LYP/aug-cc-pVTZ//B3LYP/6-31+G(d,p) level of density functional theory with the SMD solvation model and employing methylamine as a model for the side chain of lysine. Enthalpies, barrier heights, pKa's, and reduction potentials were calculated for intermediates to find the lowest energy paths. For neutral methylamine plus guanine radical and neutral methylamine radical plus guanine, the barrier for addition at C8 is ca. 10 kcal/mol lower than that for addition at C5 and C4. The barriers for water addition at C8, C5, and C4 of guanine radical are 13-20 kcal/mol higher than that for methylamine addition at C8. Further oxidation and loss of a proton leads to 8-methylaminoguanine, the methylamino analogue of 8-oxo-7,8-dihydroguanine (8-oxoG). The barrier for the addition of a second methylamine at C5 of 8-methylaminoguanine is 4.5 kcal/mol lower than that for the corresponding addition of water. Nevertheless, if the concentration of methylamine (or lysine) is very low, water addition could be competitive with methylamine addition. This would lead to comparable fractions of 8-monosubstituted-Sp and 5-8-disubstituted-Sp, in agreement with the experimental observations.


Identification of testis development and spermatogenesis-related genes in human and mouse testes using cDNA arrays.
We have constructed cDNA microarrays from the human testis large insert cDNA library, containing 9216 genes, together with several housekeeping genes. The cDNA microarrays were used to identify gene expression differences between human fetal and adult testes. Of >8700 hybridized clones, 731 exhibited significant differential expression characteristics. About 7500 genes were identified when the same cDNA microarrays were used for hybridization with cDNA probes from mouse testis, with 256 genes having significant differential expression between the age of 1-4 weeks. Among these genes, 101 were identified as critically related to testis development and possibly to spermatogenesis since they were found in both human and mouse testes, and expressed differentially at different stages of testis development. Of the 101 development-related genes, 59 full-length cDNAs have been sequenced previously, while the full-length cDNAs of the other 42 genes have not been published. We have obtained 11 full-length sequences of the 42 genes and deposited them in the GenBank. The conserved testis development-related genes found in both human and mouse testes may include genes that are likely to be involved in testicular functions, especially spermatogenesis, thus providing a basis for further functional characterization of the genes in mouse models.


The application of tooth temperature measurement in endodontic diagnosis: a review.
Teeth with vital pulp are those with an adequate vascular supply, so the circulatory status, and not sensitivity response of the pulp tissue, has been proposed to assess pulp vitality. Some experimental methods have been used to assess this; one of them is the measurement of tooth temperature, which may indicate the pulp vasculature. Some authors have found no differences between the temperatures of teeth with vital and nonvital pulp; however, others found that teeth with nonvital pulp have lower temperatures than teeth with vital pulp. Temperature measurement as a diagnostic procedure has been described with the use of thermocouple, thermistors, infrared thermography, and cholesteric liquid crystals. Many factors may influence the results of the temperature measurement procedure. For creating controlled conditions, each patient should be examined in a thermologic environment. Patients should be asked to refrain from smoking and eating/drinking for 60 minutes before the procedure. The room should be insulated and draft free, and the temperature should be maintained at 20 degrees C. It is desirable to have them lie down or sit with adequate support for the head. The use of a rubber dam is also advocated. Tooth temperature measurement as a diagnostic procedure is reviewed to include description of devices for the test, associated factors, and value of the test.


Coleoptera genome and transcriptome sequences reveal numerous differences in neuropeptide signaling between species.
Insect neuropeptides are interesting for the potential their receptors hold as plausible targets for a novel generation of pesticides. Neuropeptide genes have been identified in a number of different species belonging to a variety of insects. Results suggest significant neuropeptide variation between different orders, but much less is known of neuropeptidome variability within an insect order. I therefore compared the neuropeptidomes of a number of Coleoptera. Publicly available genome sequences, transcriptomes and the original sequence data in the form of short sequence read archives were analyzed for the presence or absence of genes coding neuropeptides as well as some neuropeptide receptors in seventeen beetle species. Significant differences exist between the Coleoptera analyzed here, while many neuropeptides that were previously characterized from Tribolium castaneum appear very similar in all species, some are not and others are lacking in one or more species. On the other hand, leucokinin, which was presumed to be universally absent from Coleoptera, is still present in non-Polyphaga beetles. The variability in neuropeptidome composition between species from the same insect order may be as large as the one that exists between species from different orders.