Chlamydophila pneumoniae and Mycoplasma pneumoniae in respiratory specimens of children with chronic lung diseases.
Persistent infection with Mycoplasma pneumoniae and Chlamydophila pneumoniae has been implicated in the progression or induction of asthma and chronic obstructive pulmonary disease. Evidence for this hypothesis has been obtained in adults either by serological methods or by direct pathogen detection using invasive procedures. We investigated nasal brush specimens and induced sputum from 38 children with stable chronic lung disease (asthma, n = 26; chronic bronchitis n = 12) and from 42 healthy controls for the presence of M pneumoniae or C. pneumoniae DNA by polymerase chain reaction (PCR) using nested primers. None of the controls but 23.6% and 10.5% of the children with lung disease had positive PCR for C pneumoniae (p = 0.001) and M pneumoniae (p = 0.044) respectively. Significantly more children with non-atopic asthma than with atopic asthma were positive for C pneumoniae or M pneumoniae (4/8 v 1/18; p = 0.018). There were no unwanted side effects from sputum induction. No correlation was found between detection of Chlamydophila and severity of lung disease. Colonisation with both organisms had occurred before adulthood in a significant proportion of children with stable chronic lung diseases. Combining nasal brush specimens with induced sputum may be a useful non-invasive method for studying the role of C pneumoniae and M pneumoniae infection in children with different chronic lung diseases.


Improved energy output levels from small-scale Microbial Fuel Cells.
This study reports on the findings from the investigation into small-scale (6.25 mL) MFCs, connected together as a network of multiple units. The MFCs contained unmodified (no catalyst) carbon fibre electrodes and for initial and later experiments, a standard ion-exchange membrane for the proton transfer from the anode to the cathode. The anode microbial culture was of the type commonly found in domestic wastewater fed with 5 mM acetate as the carbon-energy (C/E) source. The cultures were mature and acclimatised in the MFC environment for approximately 2 months before being re-inoculated in the experimental MFC units. The cathode was of the O(2) diffusion open-to-air type, but for the purposes of the polarization experiments, the cathodic electrodes were moistened with ferricyanide. The main aim of this study was to investigate the effects of connecting multiples of MFC units together as a method of scale up by using stacks and comparison of the effects of different PEM and MFC structural materials on the performance. Impedance matching (maximum-power-transfer) was achieved through calculation of total internal impedance. Three different PEM materials were compared in otherwise identical MFCs in sets of three. For individual isolated MFCs, Hyflon E87-03 was shown to produce twice, whilst E87-10 produced approximately 1.5 times the power output of the control (standard) PEM. However, when MFCs containing the E87-03 and E87-10 membranes were connected in a stack, the system suffered from severe instability and cell reversal. To study the effects of the various polymeric MFC structural materials, four small-scale units were manufactured from three different types of RP material; acrylo-butadiene-styrene coated (ABS), ABS coated (ABS-MEK) and polycarbonate (polyC). The stack of four (4) units prototyped out of polyC produced the highest power density values in polarisation experiments (80 mW/m(2)).


Telbivudine treatment started in early and middle pregnancy completely blocks HBV vertical transmission.
To evaluate the efficacy and safety of treating HBV-positive mothers with telbivudine in early and middle pregnancy to prevent mother-to-infant HBV transmission. The subject population comprised pregnant women with chronic hepatitis B (CHB; n = 188) from January 2013 to June 2015, with HBV DNA ≥1.0 × 107copies/mL and increased alanine aminotransferase levels. Groups A (n = 62) and B (n = 61) were treated with telbivudine starting at 12 weeks or 20-28 weeks after gestation, respectively. Telbivudine was discontinued at postpartum 12 weeks. Group C (n = 65) received no antiviral. All infants were vaccinated with hepatitis B immunoglobulin (200 IU) and HBV vaccine (20 with hepatitis B The maternal HBV DNA levels of the groups were compared. Mother-to-infant transmission of HBV was indicated by the presence of HBsAg in infants 7 months after birth. Before treatment, the HBV DNA levels of the 3 groups were similar. Before delivery and 12 weeks after delivery, the HBV DNA levels of groups A and B were similar, but both were significantly lower than that of group C (P < 0.01, all). No infants in groups A and B were HBsAg-positive, but the infection rate of group C was 18.4% (P < 0.01). The HBV infection rate of infants was positively associated with the HBV DNA levels of the pregnant mothers. Administration of telbivudine to HBV-infected mothers, started during early and middle pregnancy, completely blocked mother-to-infant HBV transmission. The study was registered retrospectively on Janurary 25 in 2016 at Chinese Clinical Trial Registry ( ChiCTR-OPC-16007899 ).


Comparison of the performance of different models for the interpretation of low level mixed DNA profiles.
DNA analyses from forensic casework samples commonly result in complex DNA profiles. Often, these profiles consist of multiple contributors and display multiple stochastic events such as peak height imbalance, allelic or locus drop-out, allelic drop-in, and excessive or indistinguishable stutter. This increased complexity has established a need for more sophisticated methods of DNA mixture interpretation. This study compares the effectiveness of statistical models in the interpretation of artificially created low template two person mixed DNA profiles at varying proportions and template quantities. Two binary models (combined probability of inclusion and random match probability), a semicontinuous (Lab retriever), and continuous model (STRmix™) were compared. Generally, as the sophistication of the models increases, the power of discrimination increases. Differences in discrimination often correlate to each model's ability to use observed data effectively. Binary models require static thresholds resulting in unused data and outliers that may lead to difficult or incorrect interpretation. Semicontinuous and continuous models eliminate the stochastic threshold, however Lab Retriever does not account for stochastic events beyond drop-out and drop-in leading to possible less effective use of the data. STRmix™ incorporates all stochastic events listed above into the calculation making the most effective use of the observed data.


Computerized optimization of multiple isocentres in stereotactic convergent beam irradiation.
A method for the fully computerized determination and optimization of positions of target points and collimator sizes in convergent beam irradiation is presented. In conventional interactive trial and error methods, which are very time consuming, the treatment parameters are chosen according to the operator's experience and improved successively. This time is reduced significantly by the use of a computerized procedure. After the definition of target volume and organs at risk in the CT or MR scans, an initial configuration is created automatically. In the next step the target point positions and collimator diameters are optimized by the program. The aim of the optimization is to find a configuration for which a prescribed dose at the target surface is approximated as close as possible. At the same time dose peaks inside the target volume are minimized and organs at risk and tissue surrounding the target are spared. To enhance the speed of the optimization a fast method for approximate dose calculation in convergent beam irradiation is used. A possible application of the method for calculating the leaf positions when irradiating with a micromultileaf collimator is briefly discussed. The success of the procedure has been demonstrated for several clinical cases with up to six target points.


A lively learning agenda.
It started with little, but telling, incidents. Like the time a utility company employee phoned the hospital to inquire about its cardiac package. Someone in the admitting office told him: "I'm not aware of any such plan, sir. By the way, what is a PPO anyway?" Then, about three years ago, an internal survey on managed care education confirmed an organization-wide knowledge gap in this area: Only 4 percent of directors and 7 percent of executives claimed to have a clear understanding of managed care. These were among the wake-up calls that became the impetus for a comprehensive education program at Borgess Medical Center, a 426-bed regional referral center in Kalamazoo, Michigan.


Ethics and designs: laetrile trials as an example.
The National Cancer Institute instituted a search for cancer patients who have had objective benefit from Laetrile therapy. Cases have been solicited through a nationwide mailing to > 450,000 physicians and other health professionals, through publicity in the medical and lay press, and by contact with the pro-Laetrile groups. As a result, a retrospective analysis of the medical records of patients who volunteered their cases and who believed they had objective antitumor benefit from Laetrile therapy has been carried out. Similar criteria for response that are used for any National Cancer Institute-sponsored investigational agent were used to evaluate possible Laetrile antitumor activity. This analysis was not designed to determine a response rate to Laetrile. It is uncontrolled, only positive responders were solicited, and the total number of cases treated with Laetrile was not available. It is believed that if any objective benefit for Laetrila therapy exists, then at least hints of this should be made evident by this retrospective evaluation. Data collected were used to assist in deciding if clinical trials of Laetrile are warranted.


Constitutional and environmental risk factors for cutaneous melanoma in an Italian population. A case-control study.
The aim of this study was to determine the relative risk for cutaneous melanoma associated with phenotypic and environmental variables in a population in central Italy and to assess how the combination of the different risk factors contributes to the overall risk for melanoma. We performed a case-control study of 100 patients with sporadic cutaneous melanoma and 200 controls matched for sex, age, ethnicity and residential area. Individuals were interviewed concerning pigmentary traits and sun exposure, and underwent a total body skin examination. Logistic regression models were used to evaluate the association between cutaneous melanoma and constitutional and environmental variables. The strongest risk factors were prolonged recreational sun exposure (odds ratio [OR] 5.010, 95% confidence interval [CI] 2.110-11.891), the presence of clinically atypical naevi (OR 4.916, 95% CI 2.496-9.995) and the presence of >50 common melanocytic naevi (OR 4.684, 95% CI 2.442-9.231). In addition, occupational sun exposure (OR 2.573, 95% CI 1.399-4.732), light brown hair (OR 2.336, 95% CI 1.328-4.138), high density of solar lentigos and/or actinic keratoses (OR 1.824, 95% CI 1.0-3.510) and type II, fair skin (OR 1.815, 95% CI 1.031-3.193) and blue eyes (OR 1.757, 95% CI 1.0-3.477) were each significantly associated with cutaneous melanoma risk. The combination of individual strong risk factors was associated with up to a 46-fold increase in the risk for cutaneous melanoma. Selected pigmentary traits, sun exposure and melanocytic naevi, individually and in combination, are important risk factors for cutaneous melanoma in an Italian population.


Correlation dimension estimates of human postural sway.
Human postural sway during quiet standing demonstrates a complex structured dynamics, which has been studied by applying numerous methods, such as linear system identification methods, stochastic analysis, and nonlinear system dynamics tools. Although each of the methods applied revealed some particular features of the sway data none of them have succeeded to present a global picture of the quiet stance dynamics, which probably has both stochastic and deterministic properties. In this study we have started applying ergodic theory of dynamical systems to explore statistical characteristic of the sway dynamics observed in successive trials of a subject, different subjects in an age group, and finally different age groups constituted by children, adults, and elderly subjects. Five successive 180-s long trials were performed by each of 28 subjects in four age groups at quiet stance with eyes open. Stationary and ergodic signal characteristics of five successive center of pressure time series collected from a subject in antero-posterior direction (CoPx) were examined. 97% of the trials were found to be stationary by applying Run Test while children and elderly groups demonstrated significant nonstationary behavior. On the other hand 13 out of 24 subjects were found to be nonergodic. We expected to observe differences in complexity of CoPx dynamics due to aging (Farmer, Ott, & Yorke, 1983). However linear metrics such as standard deviation and Fourier spectra of CoPx signals did not show differences due to the age groups. Correlation dimension (Dk) estimates of stationary CoPx signals being an invariant measure of nonlinear system dynamics were computed by using the average displacement method (Eckmann & Ruelle, 1985). Postural dynamics was expanded in m-dimensional space through CoPx signal by introducing optimum time delays, τcritical. 112 out of 136 stationary CoPx signals for 24 stationary subjects converged to Dk estimates. Average of Dk estimates for children and adult groups was 3.67±0.28, whereas mean of Dk estimates for elderly subjects was 4.12±0.59. Nonlinear metrics of postural sway (τcritical, msaturated, and Dk estimates) showed significant differences with respect to the age groups. Dk estimates computed from ergodic subjects' CoPx sway trajectories revealed that human quiet standing demonstrates multiple degree of freedom dynamics having a fractal structure with a considerable level of noise embedded in the signal whose characteristics is determined individually for each subject. Furthermore by using ergodic theory of complex systems, we have been able to show that the ability to independently control multiple degrees of freedom has been affected by aging.


The Flagellin-TLR5 Axis: Therapeutic Opportunities.
Motile bacteria synthesize large-sized surface structures known as flagella through the ordered polymerization of protein subunits. Flagellin, a protein of 40-60 kDa, is the principal constituent of the flagellum; each flagellum consists of approximately 20,000 flagellin molecules. An alignment of the amino acid sequences from different Gram-negative species shows a high degree of similarity in the amino and carboxy terminal domains. In contrast, the central hypervariable regions of these proteins are quite divergent. Recent work reveals that--in addition to playing a role in bacterial adhesion--monomeric flagellin, a protein component of flagellated bacteria, can also act as a soluble immunostimulatory and proinflammatory factor, activating the immune/inflammatory axis via the Toll-like receptor 5-nuclear factor-kappaB axis. Monocytes and macrophages, as well as intestinal and pulmonary epithelial cells, respond to monomeric flagellin at low concentrations. Administration of flagellin at doses comparable to or lower than that of bacterial lipopolysaccharide (endotoxin) can induce prominent local and systemic immune/inflammatory responses in vivo. Recognition of the flagellin-TLR5 pathway offers novel opportunities for the experimental therapy of various forms of shock, sepsis, acute respiratory distress syndrome, bacterial inflammation and infection. (c) Prous Science 2002. All rights reserved.


Association between pain severity and health care resource use, health status, productivity and related costs in painful diabetic peripheral neuropathy patients.
Research has shown that painful diabetic peripheral neuropathy (pDPN) is associated with worse health outcomes. However, among pDPN patients, few studies have examined the relationship between the severity of pain and health outcomes. The current project included pDPN patients (N=1506) from the 2006, 2007, and 2008 waves of the National Health and Wellness Survey. Health status (Short Form [SF]-12), work productivity (Work Productivity and Activity Impairment questionnaire), and health care resource use in the past 6 months were compared among pDPN patients with mild, moderate, and severe pain, adjusting for demographics and clinical characteristics. More than half (51.2%) of the patients reported their pain as severe, 45.2% reported moderate pain, and 3.7% reported mild pain. Those with severe pain reported significantly lower levels of health status, higher levels of work and activity impairment, and higher levels of resource use relative to the other groups. Annual per-patient costs for those with severe pain were $12,856, $3927, and $16,783 for direct, indirect, and total costs, respectively. Both direct and total costs were significantly higher in this group relative to both mild and moderate pain patients. These results suggest that pain severity contributes substantially to the health outcomes of pDPN patients and that greater resources should be allocated to the management of patients with severe pain.


The refolding of different alpha-fetoprotein variants.
The effect of glycosylation on AFP foldability was investigated by parallel quantitative and qualitative analyses of the refolding of glycosylated and nonglycosylated AFP variants. Both variants were successfully refolded by dialysis from the denatured-reduced state, attaining comparable "refolded peak" profiles and refolding yields as determined by reversed-phase HPLC analysis. Both refolded variants also showed comparable spectroscopic fingerprints to each other and to their native counterparts, as determined by circular dichroism spectroscopy. Inclusion body-derived AFP was also readily refolded via dilution under the same redox conditions as dialysis refolding, showing comparable circular dichroism fingerprints as native nonglycosylated AFP. Quantitative analyses of inclusion body-derived AFP showed sensitivity of AFP aggregation to proteinaceous and nonproteinaceous inclusion body contaminants, where refolding yields increased with increasing AFP purity. All of the refolded AFP variants showed positive responses in ELISA that corresponded with the attainment of a bioactive conformation. Contrary to previous reports that the denaturation of cord serum AFP is an irreversible process, these results clearly show the reversibility of AFP denaturation when refolded under a redox-controlled environment, which promotes correct oxidative disulfide shuffling. The successful refolding of inclusion body-derived AFP suggests that fatty acid binding may not be required for the attainment of a rigid AFP tertiary structure, contrary to earlier studies. The overall results from this work demonstrate that foldability of the AFP molecule from its denatured-reduced state is independent of its starting source, the presence or absence of glycosylation and fatty acids, and the refolding method used (dialysis or dilution).


Sickle cell trait testing and athletic participation: a solution in search of a problem?
Carriers of a single sickle cell gene mutation generally enjoy normal lifespans without serious health consequences related to their sickle cell status, but under extreme conditions such as severe dehydration and high-intensity physical activity, complications such as exertional rhabdomyolysis, splenic infarction, and papillary necrosis can occur. Recently, the National Collegiate Athletic Association (NCAA) adopted a policy that requires sickle cell solubility testing for all incoming student athletes. However, the American Society of Hematology (ASH) and other physician organizations oppose this policy. What is the basis for this controversy and how have new findings moved the field forward? I discuss herein the epidemiology, genetics, and clinical studies of sickle cell trait; review the implications of current policies regarding sickle cell trait screening and interventions for the student athlete; and examine additional areas where more information is needed.


Effects of acute maternal stress induced by predator cues on spatial learning and memory of offspring in the subterranean rodent Ctenomys talarum.
One of the main selection pressures to which animals are exposed in nature is predation, which affects a wide variety of biological traits. When the mother experiences this stressor during pregnancy and/or lactation, behavioral and physiological responses may be triggered in the offspring as well. Thus, in order to broaden and deepen knowledge on the transgenerational effects of predation stress, we evaluated how maternal stress experienced during pregnancy and/or lactation affects the spatial abilities of progeny at the onset of adulthood in the subterranean rodent Ctenomys talarum. The results showed that, contrary to what was observed in other rodent species, maternal exposure to predator cues during pregnancy and lactation did not negatively affect the spatial abilities of the offspring, even registering some minor positive effects. Concomitantly, no effects of predatory cues on physiological parameters associated with stress were observed in the progeny. This difference in results between the present study and previous works on maternal stress highlights the importance of considering the species to be evaluated (strain, age and origin-wild or captive-) and the type of stressor used (artificial or natural, intensity of exposure) in the evaluation of the possible transgenerational effects of maternal stress.


Prevalence of cytoplasmic antibodies in systemic lupus erythematosus.
Prevalence of cytoplasmic antibodies--smooth muscle antibodies (SMA), gastric parietal cell antibodies (GPA), and mitochondrial antibodies (MTA)--was evaluated in 148 normal persons and 168 patients by indirect immunofluorescent method. Their prevalence in normal persons was 0%, 2% and 0% for SMA, GPA and MTA respectively, while SMA and MTA were positive in 5.7% and 8.6% of the 35 systemic lupus erythematosus (SLE) patients respectively. The difference in the prevalence of SMA and MTA between these two groups was statistically significant. The higher prevalence of these antibodies and the occurrence of various kinds of antibodies in SLE patients support the thesis that SLE is an autoimmune phenomenon.


Endothelin-A receptor-dependent and -independent signaling pathways in establishing mandibular identity.
The lower jaw skeleton is derived from cephalic neural crest (CNC) cells that reside in the mandibular region of the first pharyngeal arch. Endothelin-A receptor (Ednra) signaling in crest cells is crucial for their development, as Ednra(-/-) mice are born with severe craniofacial defects resulting in neonatal lethality. In this study, we undertook a more detailed analysis of mandibular arch development in Ednra(-/-) embryos to better understand the cellular and molecular basis for these defects. We show that most lower jaw structures in Ednra(-/-) embryos undergo a homeotic transformation into maxillary-like structures similar to those observed in Dlx5/Dlx6(-/-) embryos, though lower incisors are still present in both mutant embryos. These structural changes are preceded by aberrant expansion of proximal first arch gene expression into the distal arch, in addition to the previously described loss of a Dlx6/Hand2 expression network. However, a small distal Hand2 expression domain remains. Although this distal expression is not dependent on either Ednra or Dlx5/Dlx6 function, it may require one or more GATA factors. Using fate analysis, we show that these distal Hand2-positive cells probably contribute to lower incisor formation. Together, our results suggest that the establishment of a 'mandibular identity' during lower jaw development requires both Ednra-dependent and -independent signaling pathways.


Evaluation of In Vivo Toxicity of Biological Nanoparticles.
Biologically derived nanoparticles such as extracellular vesicles are promising candidates for therapeutic applications. In vivo toxicity of biological nanoparticles can result in tissue or organ damage, immunological perturbations, or developmental effects but cannot be readily predicted from in vitro studies. Therefore, an essential component of the preclinical assessment of these particles for their use as therapeutics requires screening for adverse effects and detailed characterization of their toxicity in vivo. However, there are no standardized, comprehensive methods to evaluate the toxicity profile of nanoparticle treatment in a preclinical model. Here, we first describe a method to prepare bovine milk-derived nanovesicles (MNVs). These MNVs are inexpensive to isolate, have a scalable production platform, and can be modified to achieve a desired biological effect. We also describe two vertebrate animal models, mice and zebrafish, that can be employed to evaluate the toxicity profile of biologically derived nanoparticles, using MNVs as an example. Treatment-induced organ toxicity and immunological effects can be assessed in mice receiving systemic injections of MNVs, and developmental toxicity can be assessed in zebrafish embryos exposed to MNVs in embryo water. Utilizing these animal models provides opportunities to analyze the toxicity profiles of therapeutic extracellular vesicles in vivo. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Preparation of milk-derived nanovesicles Basic Protocol 2: In vivo screening for organ toxicity and immune cell profiling using mice Basic Protocol 3: In vivo developmental toxicity screening using zebrafish.


Metabolic labeling of model organisms using heavy nitrogen (15N).
Quantitative proteomics aims to identify and quantify proteins in cells or organisms that have been obtained from different biological origin (e.g., "healthy vs. diseased"), that have received different treatments, or that have different genetic backgrounds. Protein expression levels can be quantified by labeling proteins with stable isotopes, followed by mass spectrometric analysis. Stable isotopes can be introduced in vitro by reacting proteins or peptides with isotope-coded reagents (e.g., iTRAQ, reductive methylation). A preferred way, however, is the metabolic incorporation of heavy isotopes into cells or organisms by providing the label, in the form of amino acids (such as in SILAC) or salts, in the growth media. The advantage of in vivo labeling is that it does not suffer from side reactions or incomplete labeling that might occur in chemical derivatization. In addition, metabolic labeling occurs at the earliest possible moment in the sample preparation process, thereby minimizing the error in quantitation. Labeling with the heavy stable isotope of nitrogen (i.e., (15)N) provides an efficient way for accurate protein quantitation. Where the application of SILAC is mostly restricted to cell culture, (15)N labeling can be used for micro-organisms as well as a number of higher (multicellular) organisms. The most prominent examples of the latter are Caenorhabditis elegans and Drosophila (fruit fly), two important model organisms for a range of regulatory processes underlying developmental biology. Here we describe in detail the labeling with (15)N atoms, with a particular focus on fruit flies and C. elegans. We also describe methods for the identification and quantitation of (15)N-labeled proteins by mass spectrometry and bioinformatic analysis.


In vivo monitoring of focused ultrasound surgery using local harmonic motion.
The present study established the feasibility of a technique for monitoring focused ultrasound (FUS) lesion formation in vivo using localized harmonic motion (LHM) measurements. Oscillatory motion (frequencies between 50 and 300 Hz) was generated within tissues by induction of a periodic radiation force with a FUS transducer. The harmonic motion was estimated using cross correlation of RF ultrasonic signals acquired at different instances during the motion by using a confocal diagnostic ultrasound transducer. The technique was evaluated in vivo in rabbit muscle (14 locations) in an magnetic resonance (MR) imager for simultaneous ultrasound harmonic motion tracking and MR thermometry. The measured maximum amplitude of the induced harmonic motion before and after the lesion formation was significantly different for all the tested motion frequencies, and decreased between 17 and 81% depending on the frequency and location. During the FUS exposure a drop in the maximum amplitude value was observed and a threshold value could be associated to the formation of a thermal lesion. A series of controlled sonications was performed by stopping the exposure when the threshold value in LHM amplitude was reached and the presence of a thermal lesion was confirmed by MR imaging. LHM measurements were also used to perform a spatial scan of the tissues across the exposure region and the thermal lesions could be detected as a reduction in the maximum motion amplitude value at the sonication region.


Theta burst stimulation in the treatment of chronic orofacial pain: a randomized controlled trial.
Theta burst stimulation (TBS) is a modified form of high-frequency repetitive transcranial magnetic stimulation (rTMS) with promising effect in chronic pain. The aim of our double-blind, sham-controlled, parallel-group, randomized study was to assess an efficacy of intermittent TBS (iTBS) in the treatment of patients with chronic orofacial pain. Nineteen patients (twelve females) with chronic orofacial pain were prospectively included and randomly assigned to single session of an active (iTBS) or sham (intermediate TBS; imTBS) stimulation delivered to the primary motor cortex (M1) contralateral to painful side. The primary outcome was pain relief assessed using a visual analogue scale (VAS) after stimulation and at the end of two-week follow- up. The secondary outcomes were changes in the quantitative sensory testing (QST). QST set the threshold for thermal and tactile (touch) sensation in the affected facial area. Intermittent TBS, compared with the sham, showed significant improvement in VAS after stimulation, but not at the end of two-week follow-up. Regarding the secondary outcomes (QST), we failed to find any significant difference between iTBS and sham. Our findings demonstrate that iTBS of M1 transiently provides transient and modest subjective pain relief in chronic orofacial pain.


Detection of pulmonary and extrapulmonary tuberculosis patients with the 38-kilodalton antigen from Mycobacterium tuberculosis in a rapid membrane-based assay.
A rapid membrane-based serologic assay using the 38-kDa antigen from Mycobacterium tuberculosis for the diagnosis of tuberculosis (TB) was evaluated with 201 patients with pulmonary TB, 67 patients with extrapulmonary TB, 79 Mycobacterium bovis BCG-vaccinated healthy controls, and 77 non-TB respiratory patients. The overall sensitivities, specificities, and positive and negative predictive values were, respectively, 92, 92, 84, and 96% for sputum-positive TB patients; 70, 92, 87, and 79% for sputum-negative TB patients; and 76, 92, 80, and 90% for extrapulmonary-TB patients. Only 2% (1 of 44) of the healthy control BCG-vaccinated subjects gave weak positive signals in the assay, indicating that this rapid serological assay is a valuable aid in clinical diagnosis for both pulmonary and extrapulmonary TB.


Modulation of growth and differentiation of human colon carcinoma cells by histone deacetylase inhibitory trichostatins.
Histone deacetylase inhibitors such as sodium butyrate or (R)-trichostatin A {(R)-TSA; 7- [4-(dimethylamino) phenyl]-N-hydroxy-4,6-dimethyl-7-oxo-2, 4-heptadienamide} have been reported to modulate the proliferation and differentiation of certain cell types. In this study, we analyzed the effects of these agents on KM12 human colon carcinoma (HCC) cells in culture. We found that (R)-TSA induced cell flattening, inhibited anchorage-dependent and anchorage-independent growth, increased the level of the differentiation marker carcinoembryonic antigen, and increased the expression of gelsolin, a candidate tumor suppressor, in these HCC cells. Cells treated with (R)-TSA for 3 h exhibited a high degree of histone (primarily H4) acetylation. (R)-TSA exerted these effects at concentrations in the range between 0.1 to 1 mu M that are at least 1,000 times lower than those required to achieve similar effects by n-butyrate. Trichostatin C, which exhibits some histone deacetylase inhibitory activity but not the analogs (S)-TSA or (R)- or (S)-trichostatic acid, which lack histone deacetylase inhibitory activity, also showed some growth inhibitory activity. These results indicate that histone deacetylase inhibitors may lead to suppression of the transformed phenotype and enhanced differentiation in the KM12 HCC cells.


Augmentation of mouse natural killer (NK) activity by GM-1/P, a processed form of monosialoganglioside GM-1.
We describe the immunomodulatory activity of GM-1/P a processed form of GM-1 (monosialoganglioside) extracted from ox brain, purified and physically modified. We examined the effect of in vivo and in vitro treatment of GM-1/P on natural (NK) activity and its ability to induce the production of interleukin-2 (IL-2) in the mouse. In vivo treatment with GM-1/P (1 mg/Kg, i.v., day-1) resulted in a marked increase and in a change of distribution of NK activity, which was associated with lower density Percoll fractions. Marked increase was already observed at 18 hrs and then declined by day 4. In vitro treatment with GM-1/P (2 micrograms/ml) enhanced NK activity of B6 spleen cells, already after 6 hours of incubation, remaining at plateau levels within 18 hours. A role of IL-2 in this enhancement was suggested by the ability of an anti-IL-2 rabbit antiserum to abolish in vitro increased cytotoxicity. The presence of IL-2 in the supernatants of splenocytes from GM-1/P (1mg/Kg, i.v., ,day-1) treated mice stimulated with Con A or Con A plus TPA for 48 hrs was evaluated by proliferation of an IL-2 dependent CTLL cell line. GM-1/P by itself was unable to stimulate IL-2 production; however it markedly increased IL-2 production induced by Con A or Con A plus TPA.


The effect of cytochalasin D on steroid production and stress fiber organization in cultured bovine adrenocortical cells.
The effects of cytochalasin D on basal and adrenocorticotropin (ACTH)-stimulated steroid production as well as stress fiber organization by bovine adrenocortical (BAC) cells grown in monolayer culture have been investigated. Corticosteroid and pregnenolone release was determined by use of radioimmunoassay. The addition of ACTH (1 nM) produced a 30-fold increase in steroid release (principally corticosterone) in a 2 h period. Cytochalasin D (1-10 microM) had no effect on ACTH-stimulated release of corticosteroids. Basal steroid release was elevated by cytochalasin D with lower concentrations being more effective. In addition, hormonal stimulation of pregnenolone formation, the initial step in cholesterol metabolism leading to corticosteroid production, was also unaffected by cytochalasin D (1-50 microM) addition. Observation of stress fibers by fluorescence microscopy using the probe NDB - phalladin revealed that cytochalasin D (10 and 50 microM) caused an aggregation of actin-containing filaments into stellate foci within the cytoplasm. This was confirmed by electron microscopic examination of the cells. ACTH, however, had no observable effect on stress fiber organization or cell morphology. These results differ from the inhibitory effect of cytochalasin on steroid production observed using Y-1 adrenal tumor cells and rat adrenal cells in primary culture. BAC cells in contrast to Y-1 adrenal tumor cells differ in non-esterified cholesterol content and their morphological response to ACTH treatment. We suggest that these differences may influence the lack of inhibition of cytochalasin D on ACTH-stimulated steroid release.


Evaluating tests for cluster-randomized trials with few clusters under generalized linear mixed models with covariate adjustment: A simulation study.
Generalized linear mixed models (GLMM) are commonly used to analyze clustered data, but when the number of clusters is small to moderate, standard statistical tests may produce elevated type I error rates. Small-sample corrections have been proposed for continuous or binary outcomes without covariate adjustment. However, appropriate tests to use for count outcomes or under covariate-adjusted models remains unknown. An important setting in which this issue arises is in cluster-randomized trials (CRTs). Because many CRTs have just a few clusters (eg, clinics or health systems), covariate adjustment is particularly critical to address potential chance imbalance and/or low power (eg, adjustment following stratified randomization or for the baseline value of the outcome). We conducted simulations to evaluate GLMM-based tests of the treatment effect that account for the small (10) or moderate (20) number of clusters under a parallel-group CRT setting across scenarios of covariate adjustment (including adjustment for one or more person-level or cluster-level covariates) for both binary and count outcomes. We find that when the intraclass correlation is non-negligible ( ≥ $$ \ge $$ 0.01) and the number of covariates is small ( ≤ $$ \le $$ 2), likelihood ratio tests with a between-within denominator degree of freedom have type I error rates close to the nominal level. When the number of covariates is moderate ( ≥ $$ \ge $$ 5), across our simulation scenarios, the relative performance of the tests varied considerably and no method performed uniformly well. Therefore, we recommend adjusting for no more than a few covariates and using likelihood ratio tests with a between-within denominator degree of freedom.


Polymer brush guided formation of thin gold and palladium/gold bimetallic films.
This manuscript reports a new strategy to guide the chemical solution deposition of thin, microstructured metal films. The proposed strategy is based on the use of poly(2-(methacryloyloxy)ethyl ammonium chloride) (PMETAC) brushes grown via surface-initiated atom transfer radical polymerization as a template. Thin gold films have been prepared by first loading the PMETAC brushes with HAuCl(4), followed by a NaBH(4) mediated reduction to produce a PMETAC-gold nanoparticle composite film and finally an oxygen plasma treatment to remove the stabilizing polymer brush matrix and generate the desired thin gold film. The thickness of the gold films was found to scale with the thickness of the PMETAC brush template. This approach can also be extended to more complex, bimetallic films by exposing the PMETAC template successively to two different precursor salts. In this way, gradient type bimetallic palladium/gold films could be prepared.


Mapping Hot Electron Response of Individual Gold Nanocrystals on a TiO2 Photoanode.
Incorporating metal nanocrystals with semiconductor photoanodes significantly enhances the efficiency of the energy conversion in the visible range during water splitting due to the excitation of hot electrons. While extensively studied on ensemble samples, hot electron response of metal nanocrystals in a photoelectrochemical cell remains unexploited at the single-particle level. Herein, we systematically investigate hot electron response of individual single-crystalline gold nanocrystals (AuNCs) on a TiO2 photoanode during water splitting. We directly correlate the morphology of the AuNC and its plasmonic property to the efficiencies involving hot electrons with the help of single-particle dark-field microscopy and photocurrent mapping. Our results show that the efficiencies of individual AuNCs are dependent on a variety of factors including interface condition, applied bias, excitation power, incident angle, and AuNC size. Our research may shed light on optimizing the light-harvesting capability of metal/semiconductor photoanodes by providing insights into the photocatalytic processes.


Drebrin A is a postsynaptic protein that localizes in vivo to the submembranous surface of dendritic sites forming excitatory synapses.
Drebrin A is a neuron-specific, actin binding protein. Evidence to date is from in vitro studies, consistently supporting the involvement of drebrin A in spinogenesis and synaptogenesis. We sought to determine whether drebrin A arrives at the plasma membrane of neurons, in vivo, in time to orchestrate spinogenesis and synaptogenesis. To this end, a new antibody was used to locate drebrin A in relation to electron microscopically imaged synapses during early postnatal days. Western blotting showed that drebrin A emerges at postnatal day (PNd) 6 and becomes progressively more associated with F-actin in the pellet fraction. Light microscopy showed high concentrations of drebrin A in the synaptic layers of the hippocampus and cortex. Electron microscopy revealed that drebrin A in these regions is located exclusively in dendrites both neonatally and in adulthood. In adulthood, nearly all of the synaptic drebrin A is within spines forming asymmetric excitatory synapses, verified by gamma-aminobutyric acid (GABA) negativity. At PNd7, patches of drebrin A immunoreactivity were discretely localized to the submembranous surfaces of dendrites forming slight protrusions-protospines. The drebrin A sites exhibited only thin postsynaptic densities and lacked axonal associations or were contacted by axons that contained only a few vesicles. Yet, because of their immunoreactivity to the NR2B subunit of N-methyl-D-aspartate receptors and immunonegativity of axon terminals to GABA, these could be presumed to be nascent, excitatory synapses. Thus, drebrin A may be involved in organizing the dendritic pool of actin for the formation of spines and of axospinous excitatory synapses during early postnatal periods.


Antibodies to nonpolymorphic determinants of the Thy-1 molecule inhibit T cell proliferation.
The effect of anti-Thy-1 monoclonal antibodies on murine mixed lymphocyte reactions and concanavalin A-induced mitogenesis were investigated. It is demonstrated that rat antibodies against nonpolymorphic determinants of the murine Thy-1 antigen inhibited cell proliferation in the absence of complement. In contrast, antibodies against polymorphic determinants of Thy-1 had no effect on T cell activation. Inhibition of T cell proliferation did not depend on the isotype of the blocking antibody, because both IgM and IgG antibodies against monomorphic determinants were inhibitory, whereas IgM or IgG antibodies against allotypic determinants were inactive. In addition, the blocking activity could not be attributed to the xenogeneic (rat) origin of the antibodies to nonpolymorphic Thy-1 determinants, because rat anti-Thy-1.2 antibodies had no effect on cell activation. Thus, the efficacy of anti-Thy-1 antibodies as T cell inhibitors was determined by the antibody specificity. The suppressive mechanism of anti-Thy-1 antibodies was effective throughout the entire course of mixed lymphocyte reactions. Addition of antibodies at any time point during the first 90 hr of a 120-hr mixed lymphocyte culture resulted in significant suppression of the proliferative response. However, in some cases an early enhancement preceded suppression of the response. The modulation of proliferative responses by anti-Thy-1 did not result from a nonspecific mitogenic effect of the antibodies on T lymphocytes, because no effects were observed when antibodies were added to responder cells alone. These results suggest that the Thy-1 molecule, or a molecule that is located on the cell membrane in close proximity to the Thy-1 antigen, is involved in the activation of T lymphocytes.


No evidence for genomic imprinting in liver-born Down syndrome patients.
Despite numerous studies, the clinical heterogeneity of Down syndrome has no explanation. We have attempted to investigate the role of genomic imprinting in the phenotype of liveborn Down syndrome patients. Hundred fifty eight patients were investigated for parental origin of the extra chromosome 21 with standard cytogenetic analyses and with DNA plymorphic markers. The extra chromosome 21 was of paternal origin in 8 cases and of maternal origin in 150 cases. The phenotype of Down syndrome patients in whom the nondisjunction was of maternal origin, was not different from the phenotype of Down syndrome patients in whom the nondisjunction was of paternal origin. We conclude that imprinting may probably not play a role in the heterogeneity of Down syndrome phenotype.


[Therapy of chronic venous insufficiency--the domain of dermatologic ambulatory care].
By means of a graded test system for external medicaments, we optimized and standardized the local therapy of chronic venous insufficiency (CVI) in our out-patient department: allergological tests; subjective sensitivity tests of ulcus cruris venosum; comparison of ointments with powder in the therapy of ulcus cruris venosum, objective findings, applicability of the dry therapy (Heede); comparison of external drugs containing antimicrobial chemotherapeutics with those not containing these additives in the therapy of ulcus cruris venosum; comparison of ointments containing heparinoid with lipoid-in-water emulsions without admixtures in plebological therapy. We report on the therapeutic standard. Clinically 63,8% of all patients attending our dermatologic out-patient department showed CVI (stages I to IV); 38,7% were graded "venously ill" (stages III and IV). Stage I of CVI is characterized by varicous veius without discomfort. Problems of eczema are both dermatological and phlebological problems. The therapy of CVI, however, is the dermatologist's business.


Effects of DNA polymerase inhibitors on replicative and repair DNA synthesis in ultraviolet-irradiated HeLa cells.
Aphidicolin specifically inhibits eukaryotic DNA polymerase alpha, while 2',3'-dideoxythymidine 5'-triphosphate (d2TTP) inhibits DNA polymerase beta and gamma but not alpha. 1-beta-D-Arabinofuranosylcytosine 5'-triphosphate (araCTP) inhibits both DNA polymerase alpha and beta although to a different extent. Here we measured the effects of these inhibitors on repair DNA synthesis of U.V.-irradiated HeLa cells by two different methods. Firstly, aphidicolin, 1-beta-D-arabinofuranosylcytosine (araC, a precursor of araCTP) and 2',3'-dideoxythimidine (d2Thd, a precursor of d2TTP) were added directly to the culture medium. In this case, aphidicolin and araC strongly inhibited replicative DNA synthesis of HeLa cells, and they also inhibited repair synthesis after U.V.-irradiation but to a much lesser extent. In contrast, high concentrations of d2Thd inhibited repair DNA synthesis to a higher extent than replicative DNA synthesis. Secondly, the active form of inhibitor, d2TTP, was microinjection directly into cytoplasm or nuclei or U.V.-irradiated HeLa cells. Microinjection of d2TTP effectively inhibited repair synthesis. The microinjection of d2TTP, into either cytoplasm or nucleus, strongly inhibited replicative synthesis. These results might indicate that multiple DNA polymerases are involved in repair synthesis as well as in replicative synthesis.


Brain allopregnanolone in the fetal and postnatal rat in response to uteroplacental insufficiency.
Allopregnanolone suppresses central nervous system activity and has neuroprotective actions in hypoxia-induced brain injury. In pregnant sheep allopregnanolone concentrations are high during fetal life and decline rapidly after birth. We investigated brain allopregnanolone concentrations of fetal and postnatal rats derived from normal and growth restricted pregnancies. Bilateral uterine vessel ligation (or sham) was performed at gestation day 18 to induce uteroplacental insufficiency in WKY rats (n = 7-8 per group). Brain allopregnanolone was measured by radioimmunoassay at 2 study ages, gestation day 20 (n = 6 per group) and postnatal day 6 (n = 6-8 per group), from control and growth-restricted pups. Fetal brain allopregnanolone concentrations were higher in growth-restricted fetuses compared to control (p < 0.05). Allopregnanolone concentrations decreased at birth with a greater decline in growth restriction (p < 0.05). Postnatal day 6 brain allopregnanolone concentrations were lower in growth restriction (p < 0.05). Growth restriction is a potent stimulus for neurosteroid synthesis in the fetal brain in late pregnancy. The low concentrations of allopregnanolone in the growth-restricted postnatal brain suggest a delay in the capacity of the adrenal gland or brain to synthesize pregnane steroids or their precursors and may render the postnatal brain vulnerable to hypoxia-induced injury.


CEST, ASL, and magnetization transfer contrast: How similar pulse sequences detect different phenomena.
Chemical exchange saturation transfer (CEST), arterial spin labeling (ASL), and magnetization transfer contrast (MTC) methods generate different contrasts for MRI. However, they share many similarities in terms of pulse sequences and mechanistic principles. They all use RF pulse preparation schemes to label the longitudinal magnetization of certain proton pools and follow the delivery and transfer of this magnetic label to a water proton pool in a tissue region of interest, where it accumulates and can be detected using any imaging sequence. Due to the versatility of MRI, differences in spectral, spatial or motional selectivity of these schemes can be exploited to achieve pool specificity, such as for arterial water protons in ASL, protons on solute molecules in CEST, and protons on semi-solid cell structures in MTC. Timing of these sequences can be used to optimize for the rate of a particular delivery and/or exchange transfer process, for instance, between different tissue compartments (ASL) or between tissue molecules (CEST/MTC). In this review, magnetic labeling strategies for ASL and the corresponding CEST and MTC pulse sequences are compared, including continuous labeling, single-pulse labeling, and multi-pulse labeling. Insight into the similarities and differences among these techniques is important not only to comprehend the mechanisms and confounds of the contrasts they generate, but also to stimulate the development of new MRI techniques to improve these contrasts or to reduce their interference. This, in turn, should benefit many possible applications in the fields of physiological and molecular imaging and spectroscopy.


Diabetic RBC-induced oxidant stress leads to transendothelial migration of monocyte-like HL-60 cells.
Red blood cells (RBC) from patients with diabetes mellitus exhibit an increased propensity to adhere to cultured human umbilical vein endothelial cells (HUVEC) as a result of interaction of advanced glycation end products with their counter receptors, contributing to the pathogenesis of vascular complications. We determined whether the interaction of diabetic RBC with HUVEC induced cellular oxidant stress that would culminate in adherence and diapedesis of monocytes, these being initiating events in endothelial injury and atherogenesis. We show that the adherence of diabetic RBC (2% hematocrit), but not normal RBC, to HUVEC results in a fourfold increase in the production of lipid peroxides. Furthermore, diabetic RBC-induced oxidant stress causes a sixfold increase in platelet endothelial cell adhesion molecule-1 (PECAM-1) phosphorylation and doubles transendothelial migration of monocyte-like HL-60 cells; both are blocked by antioxidants and protein kinase C (PKC) inhibitors. Our results show that the adherence of diabetic RBC to endothelial cells initiates a cascade of cellular events resulting in PKC activation, causing PECAM-1 phosphorylation and concomitant transendothelial migration of monocytes. The increased diapedesis of monocytes, brought about by the interaction of diabetic RBC across vascular endothelium, may play an important role in accelerated atherosclerosis and cardiovascular disease in diabetics.


Identification of two new genes, mukE and mukF, involved in chromosome partitioning in Escherichia coli.
We have previously reported that the MukB protein is essential for chromosome partitioning in Escherichia coli and that mukB mutants produce anucleate cells and are temperature-sensitive for colony formation. The mukB gene maps at 21 min on the E. coli chromosome and smtA-mukF-mukE-mukB genes might comprise an operon, which is transcribed in a clockwise direction. Here, we report that mukF and mukE null mutants are both temperature-sensitive for colony formation and produce anucleate cells even at the permissive temperature. These phenotypes are the same as those observed in the mukB null mutant. The primary sequence of MukF includes a leucine zipper structure and an acidic domain. Mutational analysis revealed that both are required for MukF function. When the MukF protein was overproduced in the wild-type strain, anucleate cells were produced. In contrast, overproduction of either MukE or MukB did not cause the defect. In null mutants for the mukF, mukE, and mukB genes, the synchronous initiation of chromosome replication was not affected. The mini-F plasmid was as stably maintained in these mutants as in the wild-type strain. These results indicate that the MukF, MukE, and MukB proteins are involved in the chromosome partitioning steps, but are not required for mini-F plasmid partitioning.


DNA methylation patterns are related to co-morbidity status and circulating C-reactive protein levels in the nursing home elderly.
Elderly residents of long-term care commonly exhibit a number of age-related health deficits, including impaired physical and cognitive function, chronic illnesses, and chronic inflammation. Given our previous data relating the phenotype and function of innate and adaptive leukocytes from the nursing home elderly to chronic conditions and inflammatory biomarkers, we hypothesized that these factors would influence the regulatory programming of immune cells, thereby contributing to immune dysfunction. Since DNA methylation represents both an important regulatory mechanism of cells as well as a biomarker of health and disease, we sought to characterize the methylome of peripheral blood mononuclear cells (PBMCs) from the nursing home elderly (n=23; 82-98years old, 70% female), and compare these patterns to pathological factors such as dementia, co-morbidity score and frailty, and immune-related factors such as serum C-reactive protein (CRP) and cytokine levels and varicella-zoster virus (VZV) vaccine responsiveness. We show that the most significant changes in DNA methylation levels occurred in relation to co-morbidity score, including one site, cg07725579 (FDR-adjusted p<0.05; closest gene, SIRBP2), and nine DNA methylation regions (Stouffer's p<0.05). DNA methylation age, although not strongly correlated with chronological age, was positively correlated with serum CRP levels (p=0.007), and negatively correlated with vaccine responsiveness (p=0.035). To our knowledge, this study is one of the first to describe associations of DNA methylation patterns with pathological and immune-related factors in residents of long-term care, and may provide important clues pertaining to immune cell dysfunction near the end of life.


Same place, different time, head up: Multiple antipredator responses to a recolonizing apex predator.
Prey adjust their antipredator behavioral tactics to minimize the risk of an encounter with predators. Spatiotemporal responses of prey to predators have been reported, but the nature of antipredator response is not ubiquitous and it is the object of increasing interest, especially considering the recent recovery of large carnivores in Europe, and the potential for behavioral antipredator responses to elicit consequences at the ecosystem level. We have tested multiple antipredator responses by fallow deer Dama dama to wolf Canis lupus in a Mediterranean protected area recently recolonized by this apex predator. Through intensive camera trapping, we tested for temporal and spatial association between predator and prey, and we have also studied deer vigilance in forest habitats where focal observations are usually impossible. Wolf detection rates were spatially associated with those of fallow deer. Accordingly, no evidence was found for fallow deer avoiding sites with higher predator detection rates. Temporal activity patterns were significantly different between the 2 species, with the wolf being mainly nocturnal whereas fallow deer was active especially during daylight. A comparison with a preliminary study strongly suggests an increase in the diurnal activity of fallow deer along with the stabilization of wolf presence in the area. Both the rate and the duration of vigilance of female fallow deer increased with the local frequency of wolf activity. We suggest an antipredator response based on temporal-rather than spatial-avoidance, as well as increased vigilance.


Improving emotion recognition in anorexia nervosa: An experimental proof-of-concept study.
Previous research has found increasing evidence for difficulties in emotion recognition ability (ERA) and social cognition in anorexia nervosa (AN), and recent models consider these factors to contribute to the development and maintenance of the disorder. However, there is a lack of experimental studies testing this hypothesis. Therefore, the present proof-of-concept study examined whether ERA can be improved by a single session of a computerized training in AN, and whether this has short-term effects on eating disorder symptoms. Forty inpatients (22.20 ± 7.15 years) with AN were randomly assigned to receive a single session of computerized training of ERA (TERA) or a sham training (training the recognition of different types of clouds). ERA, self-reported eating disorder symptoms, and body mass index (BMI) were assessed within 3 days before and after training. After training, both groups showed improved ERA, reduced self-reported eating disorder symptoms, and an increased BMI. As compared to patients in the control group, patients who received TERA showed greater improvements in ERA and self-reported eating disorder symptoms. ERA can be effectively trained in patients with AN. Moreover, short-term improvements in self-reported eating disorder symptoms provide tentative support for the hypothesis that difficulties in ERA contribute to the maintenance of AN, and that specific trainings of ERA hold promise as an additional component in AN treatment. Future studies are needed to replicate these findings in larger samples, and to investigate long-term effects and transfer into real-world settings.


[Clinical and genetic study of an infant with Alagille syndrome: identification of a novel chromosomal interstitial deletion including JAG1 gene].
Alagille syndrome (ALGS) is an autosomal dominant disease affecting multiple systems including the liver, heart, skeleton, eyes, kidneys and face. This paper reports the clinical and genetic features of an infant with this disease. A 3-month-and-10-day-old female infant was referred to the hospital with jaundiced skin and sclera for 3 months. Physical examination revealed wide forehead and micromandible. A systolic murmur of grade 3-4/6 was heard between the 2th and 3th intercostal spaces on the left side of the sternum. The abdomen was distended, and the liver palpable 3 cm under the right subcostal margin with a medium texture. Serum biochemistry analysis revealed abnormal liver function indices, with markedly elevated bilirubin (predominantly direct bilirubin), total bile acids (TBA) and gamma-glutamyl transpeptidase (GGT). Atrial septal defect and pulmonary stenosis were detected on echocardiography. Next generation sequencing detected entire deletion of the JAG1 gene, and then chromosomal microarray analysis revealed a novel interstitial deletion of 3.0 Mb in size on chr20p12.3p12.2, involving JAG1 gene. The child had special facial features, heart malformations, and cholestasis, and based on the genetic findings, ALGS was definitively diagnosed. Thereafter, symptomatic and supportive treatment was introduced. Thus far, the infant had been followed up till his age of 11 months. The hyperbilirubinemia got improved, but GGT and TBA were persistently elevated, and the long-term outcome needs to be observed. This study extended the JAG1 mutation spectrum, and provided laboratory evidences for the diagnosis and treatment of the patient, and for the genetic counseling and prenatal diagnosis in the family.


Clostridium botulinum type E toxin: effect of pH and method of purification on molecular weight.
The toxin of Clostridium botulinum type E was isolated from intact cells and from toxic culture filtrates by column chromatography at three pH values, 4.5, 5.3, and 6.0. At pH 6.0 and 5.3, the isolated toxin was in a form with a molecular weight (MW) of 86,000. This toxin was homogeneous on polyacrylamide gel electrophoresis and gel filtration and had an optical density ratio, 280 nm/260 nm, greater than 2.0. It did not dissociate at higher pH levels, but was dissociated into nontoxic components of approximately 12,000 MW when reduced and alkylated in the presence of 6 M guanidine hydrochloride. At pH 4.5, smaller amounts of an impure toxic moiety with a MW of 12,000 were found. After storage for 6 months, the 86,000-MW moiety had lost 60% of its lethality. Gel filtration revealed that the bulk of the toxicity was associated with a component having a MW of 150,000. Toxic components with MW of 12,000 and over 200,000 were also found. The toxin appears to polymerize or aggregate when in a pure form, so that most, if not all, of the MW previously reported for the toxin may belong to different polymers of a monomer with a MW of 12,000 or less. Treatment of the 86,000-MW toxin with trypsin resulted in an 18- to 128-fold increase in lethality, but no detectable change in MW.


Genomic Evidence of In-Flight Transmission of SARS-CoV-2 Despite Predeparture Testing.
Since the first wave of coronavirus disease in March 2020, citizens and permanent residents returning to New Zealand have been required to undergo managed isolation and quarantine (MIQ) for 14 days and mandatory testing for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). As of October 20, 2020, of 62,698 arrivals, testing of persons in MIQ had identified 215 cases of SARS-CoV-2 infection. Among 86 passengers on a flight from Dubai, United Arab Emirates, that arrived in New Zealand on September 29, test results were positive for 7 persons in MIQ. These passengers originated from 5 different countries before a layover in Dubai; 5 had negative predeparture SARS-CoV-2 test results. To assess possible points of infection, we analyzed information about their journeys, disease progression, and virus genomic data. All 7 SARS-CoV-2 genomes were genetically identical, except for a single mutation in 1 sample. Despite predeparture testing, multiple instances of in-flight SARS-CoV-2 transmission are likely.


Targeting chemotherapy for malignant brain tumor using thermosensitive liposome and localized hyperthermia.
Thermosensitive liposomes are microscopic vesicles that can contain drugs and release them effectively in response to hyperthermia. To deliver an antitumor drug specifically to brain tumor, the authors used thermosensitive liposomes containing cis-diamminedichloroplatinum (CDDP) in conjunction with localized brain heating. The authors then investigated the antitumor effect on rat malignant glioma. Rous sarcoma virus-induced malignant glioma cells were transplanted into the brains of Fisher rats. Ten days after tumor inoculation, the rats were assigned to one of six treatment groups: control, free CDDP, hyperthermia, free CDDP + hyperthermia, liposomes containing CDDP (CDDP-liposome), and CDDP-liposome + hyperthermia. Liposomes containing CDDP or free CDDP were injected via the tail vein. Brain tumor heating was administered by means of a radiofrequency antenna designed at our institute. The rats treated with CDDP-liposome + hyperthermia had the longest survival time and the tumor CDDP level of this group was the highest when compared to the other groups. Histopathological examination showed that tumor cells were necrotized but surrounding normal brain tissue remained undamaged. On the basis of these findings we suggest that the combination of thermosensitive liposome and localized hyperthermia may better focus antitumor drugs to the tumor, providing a significantly greater antitumor effect.


Synthesis and evaluation of 1,3,4-oxadiazole derivatives for development as broad-spectrum antibiotics.
The reality and intensity of antibiotic resistance in pathogenic bacteria calls for the rapid development of new antimicrobial drugs. In bacteria, trans-translation is the primary quality control mechanism for rescuing ribosomes arrested during translation. Because trans-translation is absent in eukaryotes but necessary to avoid ribosomal stalling and therefore essential for bacterial survival, it is a promising target either for novel antibiotics or for improving the activities of the protein synthesis inhibitors already in use. Oxadiazole derivatives display strong bactericidal activity against a large number of bacteria, but their effects on trans-translation were recently questioned. In this work, a series of new 1,3,4-oxadiazole derivatives and analogs were synthesized and assessed for their efficiency as antimicrobial agents against a wide range of gram-positive and gram-negative pathogenic strains. Despite the strong antimicrobial activity observed in these molecules, it turns out that they do not target trans-translation in vivo, but they definitely act on other cellular pathways.


Characterization of the anti-CD22 targeted therapy, moxetumomab pasudotox, for B-cell precursor acute lymphoblastic leukemia.
Moxetumomab pasudotox is a second-generation recombinant immunotoxin against CD22 on B-cell lineages. Antileukemic activity has been demonstrated in children with chemotherapy-refractory B-cell precursor acute lymphoblastic leukemia (BCP-ALL), with variable responses. Here, we report in vitro and in vivo evaluation of moxetumomab pasudotox treatment of human cell lines and patient-derived cells as a preliminary study to understand characteristics of sensitivity to treatment. Binding, internalization, and apoptosis were evaluated using fluorescently tagged moxetumomab pasudotox. Studies in NOD-scid IL2Rgnull mice showed a modest survival benefit in mice engrafted with 697 cells but not in NALM6 or the two patient-derived xenograft models.


The factor structure of the BDI in facial pain and other chronic pain patients: a comparison of two models using confirmatory factor analysis.
1) To compare two measurement models of the BDI in chronic pain sufferers to see which provides the better fit; 2) to assess whether model fit differs for a facial pain sample compared to a sample of pain sufferers attending a multidisciplinary pain clinic; and 3) to establish which affective and somatic sub-scales of the BDI could be used in chronic pain research. Two groups of chronic pain sufferers, a facial pain group, and a group attending a multidisciplinary pain clinic completed self-report questionnaires on pain (Multidimensional Pain Inventory), depression (BDI), and measures of anxiety and depression-related pain cognitions (the Spielberger State-Trait Anxiety Inventory and the Pain Cognitions Questionnaire). The measurement models of the BDI were tested using LISREL structural equation modelling and their construct validity examined using partial correlation analysis. A total of 173 people attending a multidisciplinary pain clinic and 157 patients attending a facial pain clinic completed self-report measures of pain and mood prior to their respective clinical consultations. The model offered by Novy et al. (containing one affective factor 'Negative-attitude suicide' and two somatic factors 'Performance difficulty' and 'Physiological manifestations') fitted both pain groups better than the model offered by Williams and Richardson (containing one affective factor 'Self-reproach', one somatic factor 'Somatic disturbance' and one factor with a mixture of both affective and somatic items 'Sadness about health'). However, when the factors were allowed to correlate in the latter model, both models were broadly equivalent. The two measurement models adequately fitted data in both pain samples when the factors were allowed to intercorrelate in the Williams and Richardson model. Both the affective scales offered by both models could be used in future research, although the somatic factor offered by the Williams and Richardson model offered much higher levels of internal reliability than either of those offered in the Novy et al. model. The findings are discussed in relation to the issue of depression in chronic pain.


The Molecular Basis of G Protein-Coupled Receptor Activation.
G protein-coupled receptors (GPCRs) mediate the majority of cellular responses to external stimuli. Upon activation by a ligand, the receptor binds to a partner heterotrimeric G protein and promotes exchange of GTP for GDP, leading to dissociation of the G protein into α and βγ subunits that mediate downstream signals. GPCRs can also activate distinct signaling pathways through arrestins. Active states of GPCRs form by small rearrangements of the ligand-binding, or orthosteric, site that are amplified into larger conformational changes. Molecular understanding of the allosteric coupling between ligand binding and G protein or arrestin interaction is emerging from structures of several GPCRs crystallized in inactive and active states, spectroscopic data, and computer simulations. The coupling is loose, rather than concerted, and agonist binding does not fully stabilize the receptor in an active conformation. Distinct intermediates whose populations are shifted by ligands of different efficacies underlie the complex pharmacology of GPCRs.


Molecular Modeling and Docking of Aquaporin Inhibitors to Reveal New Insights into Schistosomiasis Treatment.
Schistosomiasis (snail fever/bilharzia), a disease caused by parasitic flatworms (schistosomes), infects millions of people worldwide. Aquaporins from these organisms were found to be a potent drug target. We investigate the possible mechanism of inhibition of Aquaporin (AQP) from S.mansoni by 5 drug molecules (Praziquantel, Metrifonate, Artimisinin, Albendazole, and Amoscanate). 3D molecular structure of Aquaporin was obtained through homology modeling and further protein-ligand docking and MD simulation were performed. VAL-75, ASN-91, ALA-220, ASN-222, ARG-225 amino acids were found to play crucial role in ligand binding. TRP-71 and other important residues play major role in hydrophobic interactions stabilizing protein-ligand complexes. We hope that this study (with the newly identified aquaporin target) will support the development of structure and pharmacophore-based novel S. mansoni drugs to control and curb Schistosomiasis.


Management and outcome of patients with acute ischemic stroke and tandem carotid occlusion in the ESCAPE-NA1 trial.
The optimal treatment and prognosis for stroke patients with tandem cervical carotid occlusion are unclear. We analyzed outcomes and treatment strategies of tandem occlusion patients in the ESCAPE-NA1 trial. ESCAPE-NA1 was a multicenter international randomized trial of nerinetide versus placebo in 1105 patients with acute ischemic stroke who underwent endovascular treatment. We defined tandem occlusions as complete occlusion of the cervical internal carotid artery (ICA) on catheter angiography, in addition to a proximal ipsilateral intracranial large vessel occlusion. Baseline characteristics and outcome parameters were compared between patients with tandem occlusions versus those without, and between patients with tandem occlusion who underwent ICA stenting versus those who did not. The influence of tandem occlusions on functional outcome was analyzed using multivariable regression modeling. Among 115/1105 patients (10.4%) with tandem occlusions, 62 (53.9%) received stenting for the cervical ICA occlusion. Of these, 46 (74.2%) were stented after and 16 (25.8%) before the intracranial thrombectomy. A modified Rankin Score (mRS) of 0-2 at 90 days was achieved in 82/115 patients (71.3%) with tandem occlusions compared with 579/981 (59.5%) patients without tandem occlusions. Tandem occlusion did not impact functional outcome in the adjusted analysis (OR 1.5, 95% CI 0.95 to 2.4). Among the subgroup of patients with tandem occlusion, cervical carotid stenting was not associated with different outcomes compared with no stenting (mRS 0-2: 75.8% vs 66.0%, adjusted OR 2.0, 95% CI 0.8 to 5.1). Tandem cervical carotid occlusion in patients with acute large vessel stroke did not lower the odds of good functional outcome in our study. Functional outcomes were similar irrespective of the management of the cervical ICA occlusion (stenting vs not stenting).


Differential lateralization of hippocampal connectivity reflects features of recent context and ongoing demands: an examination of immediate post-task activity.
Neuroimaging studies have shown that task demands affect connectivity patterns in the human brain not only during task performance but also during subsequent rest periods. Our goal was to determine whether ongoing connectivity patterns during rest contain information about both the current rest state, as well as the recently terminated task. Our experimental design consisted of two types of active tasks that were followed by two types of low-demand rest states. Using this design, we examined whether hippocampal functional connectivity during wakeful rest reflects both features of a recently terminated task and those of the current resting-state condition. We identified four types of networks: (i) one whose connectivity with the hippocampus was determined only by features of a recently terminated task, (ii) one whose connectivity was determined only by features of the current resting-state, (iii) one whose connectivity reflected aspects of both the recently terminated task and ongoing resting-state features, and (iv) one whose connectivity with the hippocampus was strong, but not affected by any external factor. The left and right hippocampi played distinct roles in these networks. These findings suggest that ongoing hippocampal connectivity networks mediate information integration across multiple temporal scales, with hippocampal laterality moderating these connectivity patterns.


Postischemic hyperemia in subjects with lower limbs obstructive arteriopathy: role of PGI2 and endothelin.
The physiological basis of postischemic hyperemia is not yet fully understood. The present study investigated the effects of pharmacologic manipulation of the prostaglandin system on local hemodynamics. Strain-gauge plethysmography was used to study 8 normal subjects and 9 patients with obliterating arterial disease of the lower limbs. Hemodynamic evaluations were performed before treatment, after seven days of low-dose acetylsalicylic acid (100 mg/day) to inhibit platelet thromboxane synthesis, and after acute infusion of 1 g of acetylsalicylic acid to inhibit endothelial prostacyclin synthesis. In patients with arterial disease, the hemodynamic study was also carried out after infusion of iloprost, a synthetic prostacyclin analogue. Acute infusion of acetylsalicylic acid significantly reduced basal blood flow in normal subjects, but not in patients with arterial disease. In the latter group, iloprost affected neither basal nor maximal postischemic flow. The study also evaluated the role of endothelin in musculocutaneous hemodynamic regulation, both in physiological conditions and in atherosclerosis. This part of the study addressed the possibility that the hemodynamic effects of vasodilator prostanoids like prostacyclin might affect endothelin release in vivo. During reactive hyperemia, plasma endothelin was reduced in normal subjects (-1.02 pg/mL, 95% CI: -2.23, 0.08), but not in patients with atherosclerosis (-0.35 pg/mL, 95% CI: -1.45, 0.75). In both groups, plasma endothelin was not affected by aspirin. These findings confirm the role of prostacyclin in local hemodynamic regulation. In the normal subject, musculocutaneous blood flow seems to depend at least in part on the action of vasodilator prostanoids and endothelin. This is not the case in patients with arterial disease, in whom plasma endothelin does not seem to be affected by postischemic changes in blood flow. A possible explanation for this difference could be alteration of the endothelial function in patients with arterial disease, related to the functional and structural characteristics of the artery wall in atherosclerosis.


Polyamidoamine dendrimer-mediated hydrogel for solubility enhancement and anti-cancer drug delivery.
The application of hydrogels for anti-cancer drug delivery has garnered considerable interest in the medical field. Current cancer treatment approaches, such as chemotherapy and radiation therapy, often induce severe side effects, causing significant distress and substantial health complications to patients. Hydrogels present an appealing solution as they can be precisely injected into specific sites within the body, facilitating the sustainable release of encapsulated drugs. This localized treatment approach holds great potential for reducing toxicity levels and improving drug delivery efficacy. In this study we developed a hydrogel delivery system containing polyamidoamine (PAMAM) dendrimer and polyethylene glycol (PEG) for solubility enhancement and sustained delivery of hydrophobic anti-cancer drugs. The three selected model drugs, e.g. silibinin, camptothecin, and methotrexate, possess limited aqueous solubility and thus face restricted application. In the presence of vinyl sulfone functionalized PAMAM dendrimer at 45 mg/mL concentration, drug solubility is increased by 37-fold, 4-fold, and 10-fold for silibinin, camptothecin, and methotrexate, respectively. By further crosslinking of the functionalized PAMAM dendrimer and thiolated PEG, we successfully developed a fast-crosslinking hydrogel capable of encapsulating a significant payload of solubilized cancer drugs for sustained release. In water, the drug encapsulated hydrogels release 30%-80% of their loads in 1-4 days. MTT assays of J82 and MCF7 cells with various doses of drug encapsulated hydrogels reveal that cytotoxicity is observed for all three drugs on both J82 and MCF7 cell lines after 48 h. Notably, camptothecin exhibits higher cytotoxicity to both cell lines than silibinin and methotrexate, achieving up to 95% cell death at experimental conditions, despite its lower solubility. Our experiments provide evidence that the PAMAM dendrimer-mediated hydrogel system significantly improves the solubility of hydrophobic drugs and facilitates their sustained release. These findings position the system as a promising platform for controlled delivery of hydrophobic drugs for intratumoral cancer treatment.


HAP2/GCS1: Mounting evidence of our true biological EVE?
Sex has consequences-indeed, where would we be without it? Yet for all its importance, remarkably little is known about how sex evolved, why it has persisted, or even what mechanisms allow sperm-egg fusion to occur. Fortunately, answers to these questions are beginning to emerge with studies of hapless 2/generative cell specific1 (HAP2/GCS1), a molecular machine that promotes gamete fusion in organisms ranging from protists to flowering plants and insects. In studies by Fedry and colleagues, key structural features of the HAP2 protein are revealed for the first time, lending new insights into its mode of action and reinforcing its relationship to viral proteins that accomplish a similar task and may be intimately linked to the origins of cell-cell fusion events (including sexual reproduction) across evolutionary time.


[Study of population dissociation of collection strains of Erwinia carotovora].
Unusual dissociants of Erwinia carotovora subsp. atroseptica (EAT) population that emerge with high frequency (18-94%) during long-term storage of cultures were found. The dissociants (Poe-phenotype) differ from parental forms by slow growth and long development of the colour of colonies on EMB agar though they are not Lac-mutants. More than 60% of Poe-mutants of strain EAT 39A are characterized by 8- and 150-fold decrease of stability to erythromycin and oleandomicin, correspondingly. They are also sensitive to colicin-like carotovoricin, which is induced by mytomicin C, to carotovoricin of initial strain EAT 39A and to autocin, which they produce themselves during induction. A multiple phenotype of these dissociants (dissociants of the second type) is labeled: PoeOmSErSBnSAu. Its appearance as well as the appearance of pure Poe-variants is not associated with usual mutations and a loss of residential plasmids by cells. Population dissociants of the both types--Poe and PoeApSOmS--were also obtained from E. carotovora subsp. carotovora (J2) during growth at supraoptimal temperature. Poe-variants are more often observed in representatives of subspecies atroseptica than carotovora and can be useful when studying pathogeneity of these practically important erwinias.


Blackwater fever in southern Vietnam: a prospective descriptive study of 50 cases.
We prospectively studied 50 Vietnamese patients with blackwater fever (BWF). All patients had fever and hemoglobinuria, 40 (80%) were jaundiced, 25 (50%) had hepatomegaly, 15 (34%) had splenomegaly, and 9 (18%) had hepatosplenomegaly. Twenty-one patients (42%) had impaired renal function, with creatinine clearances of < 50 mL/min/m2; however, only four (8%) developed oliguric renal failure, three (6%) of whom required dialysis. Forty-four patients (88%) developed anemia, which was severe (hematocrit, < 20% in 32 (64%). One patient died, representing a death rate for this once-feared disease that is considerably lower than that reported by earlier investigators. BWF was associated with quinine ingestion in 28 patients (56%), glucose-6-phosphate dehydrogenase (G6PD) deficiency in 27 (54%), and concurrent malaria infection in 16 (32%). There was no statistically significant difference in the severity of BWF associated with each of these three factors, as assessed by creatinine clearance and the hematocrit value on admission and by the number of units of blood transfused. There was considerable overlap in the occurrence of G6PD deficiency, quinine ingestion, and malaria, suggesting that these factors may interact and that it may not be justifiable to regard hemoglobinuria caused by G6PD deficiency as a separate syndrome.


A treatment applying a biomechanical device to the feet of patients with knee osteoarthritis results in reduced pain and improved function: a prospective controlled study.
This study examined the effect of treatment with a novel biomechanical device on the level of pain and function in patients with knee OA. Patients with bilateral knee OA were enrolled to active and control groups. Patients were evaluated at baseline, at 4 weeks and at the 8-week endpoint. A novel biomechanical device was individually calibrated to patients from the active group. Patients from the control group received an identical foot-worn platform without the biomechanical elements. Primary outcomes were the WOMAC Index and ALF assessments. There were no baseline differences between the groups. At 8 weeks, the active group showed a mean improvement of 64.8% on the WOMAC pain scale, a mean improvement of 62.7% on the WOMAC function scale, and a mean improvement of 31.4% on the ALF scale. The control group demonstrated no improvement in the above parameters. Significant differences were found between the active and control groups in all the parameters of assessment. The biomechanical device and treatment methodology is effective in significantly reducing pain and improving function in knee OA patients.The study is registered at clinicaltrials.gov, identifier NCT00457132, http://www.clinicaltrials.gov/ct/show/NCT00457132?order=1.


Race, Vigilant Coping Strategy, and Hypertension in an Integrated Community.
Vigilant coping refers to individuals who, potentially as a result of experiencing discrimination in the past, proactively prepare for the possibility that they will be discriminated against or mistreated because of their race. The extent to which vigilant coping is linked with hypertension, a highly prevalent condition with well-documented racial/ethnic disparities, remains largely unknown. We performed a cross-sectional analysis of data from the Exploring Health Disparities in Integrated Communities (EHDIC) study-Southwest Baltimore (n = 715). We used a series of multivariate logistic regression models to evaluate vigilance as a potential mediator or moderator of the association between race, discrimination, and hypertension within the context of a racially integrated community. There was no difference in prevalence of hypertension between African-Americans and Whites in this sample (66.8 vs. 66.2; P = 0.86). A higher proportion of African-Americans reported discrimination (41.1 vs. 22.9; P < 0.0001) and vigilance (67.3 vs. 46.9; P < 0.0001) compared to Whites; however, neither measure was associated with hypertension. In stratified analyses, bothersome discrimination alongside vigilant coping was associated with higher odds of hypertension among African-Americans and lower odds among Whites. Overall, prevalence of hypertension was high and there were no racial differences in this racially integrated community. Discrimination, vigilance, and hypertension were differently associated among African-Americans and Whites. Studying lived experiences in integrated communities may illuminate how structural inequalities impact the health of African-Americans more broadly. Further, raising awareness of social factors as they relate to hypertension may help to inform clinical management of low-income patients.


Neutropenia in Congenital and Adult Babesiosis.
Anemia and thrombocytopenia are expected hematologic abnormalities in patients with acute babesiosis, whereas neutropenia (defined as an absolute neutrophil count of ≤1,800 neutrophils/μL for adults and <1,200 neutrophils/μL for infants) is not usually considered a feature of this infection. We studied the frequency with which neutropenia occurs in congenital and adult cases of babesiosis. The frequency of neutropenia in cases of congenital babesiosis was determined based on a literature review and on the findings in an unreported case. The frequency of neutropenia in adult patients was assessed based on a review of the medical records of 51 patients who were diagnosed with babesiosis between 2010 and 2013 at two medical centers in the Northeastern United States. Four (80%; 95% confidence interval [CI], 36%-98%) of five infants with congenital babesiosis whose neutrophil count was reported were neutropenic. Among 51 adult cases with babesiosis, 11 (22%; 95% CI, 12%-35%) were neutropenic on clinical presentation, and seven others developed neutropenia over the next 1 to 21 days. Thus, a total of 18 (35%; 95% CI, 24%-49%) of the adult patients with babesiosis had neutropenia. Neutropenia appears to be a common finding in infants with congenital babesiosis and is also observed not infrequently in adults with this infection.


Feasibility of home-based dietetic intervention to improve the nutritional status of older adults post-hospital discharge.
To determine if a model of home-based dietetic care improves dietary intake and weight status in a specific group of older adults post-hospitalisation. The Department of Veterans' Affairs clients aged 65 years and over were recruited from hospitals in a regional area of New South Wales, Australia (n = 32 men, n = 36 women). Nutritional status was assessed at home at baseline (within two weeks post-discharge) and three months post-discharge using a diet history, a food frequency checklist and Mini Nutritional Assessment (MNA). Personalised dietary advice was provided by a single dietitian according to participants' nutritional status. Mean body weight improved significantly (P = 0.048), as well as mean MNA score (21.9 ± 3.5 vs 25.2 ± 3.1) (P < 0.001). Mean energy, protein and micronutrient intakes were adequate at baseline and three months, except for vitamin D. At three months, the underweight group (body mass index (BMI) < 23 kg/m2 ) had significantly higher mean protein intake per body weight (1.7 ± 0.4 g/kg) compared to those who were a desirable weight (BMI 23-27 kg/m2 ) (1.4 ± 0.3 g/kg) or overweight (BMI>27 kg/m2 ) (1.1 ± 0.3 g/kg) peers (P < 0.001). There was significant improvement in energy intake contributed from oral nutrition supplements (+95.5 ± 388.2 kJ/day) and milk (+259.6 ± 659.8 kJ/day). Dietetic intervention improved nutritional status 3 months after hospital discharge in older adults living in the community.


Work stress and burnout among emergency physicians: a systematic review of last 10 years of research.
First, to provide a synthesis and analysis of available scientific literature regarding the level of work stress and burnout among emergency physicians. Second, to identify the effect of the specific work situation-related factors. A systematic search was performed in NCBI PubMed and Embase. Comparative primary studies, both systematic review and cross-sectional, quantifying burnout in emergency physicians were included. Only studies published between 2011 and 2022 were retained. Synonym sets were compiled for the search key for 'burnout & stress', 'emergency', 'physician' and 'burnout & posttraumatic stress disorder'. Thirty-five papers were retained for further research. Emergency physicians scored significantly higher for all dimensions of burnout compared to other healthcare professions. Significant correlations for burnout were found with work characteristic and organizational factors. Critical incidents and aggression were identified as the most important acute work characteristics and organizational factors impacting emergency physician's mental wellbeing including the development of posttraumatic stress disorder. Moreover, personal factors such as age, personality, and coping strategies also play an important role in the development of burnout as well as work-related trauma. Available studies show that emergency physicians report higher scores of emotional exhaustion and depersonalization when compared to other healthcare professionals. Work characteristics contribute to this, but work-related traumatic incidents and aggression are important determinants. Personal characteristics such as age, personality type D, previous experiences and coping strategies seem to be determining factors likewise. Emergency physicians showed a high risk for developing burnout and work stress-related problems.


Assessment of the effectiveness of aircraft noise regulation.
A fundamental goal of aircraft noise regulation is control of the prevalence of noise-induced annoyance in airport communities. A common regulatory strategy is to identify values of long-term, time-weighted average aircraft noise exposure that may not be exceeded in the vicinity of airports without certain consequences. Noise exposure per se is neither the sole cause nor a perfect predictor of the annoyance of aircraft noise, however. Regulatory limitation of noise levels to certain values of favored noise metrics may therefore provide the appearance, rather than the substance, of a solution to problems of community reaction to aircraft noise. Response bias, as identified by Green and Fidell (1991), is a factor that exerts about as much influence on the observed prevalence of annoyance in communities as cumulative noise exposure. The importance of formal consideration of response bias in assessments of the adequacy of regulatory enforcement levels is addressed.


Quantitative determination and validation of 17 cannabinoids in cannabis and hemp using liquid chromatography-tandem mass spectrometry.
The increase in production of cannabis for medical and recreational purposes in recent years has led to a corresponding increase in laboratories performing cannabinoid analysis of cannabis and hemp. We have developed and validated a quantitative liquid chromatography-tandem mass spectrometry (LC-MS/MS) method that is simple, reliable, specific, and accurate for the analysis of 17 cannabinoids in cannabis and hemp. Liquid-solid sample extraction coupled with dilution into a calibration range from 10 to 10,000 ng/mL and LC-MS/MS analysis provides quantification of samples ranging from 0.002 to 200 mg/g (0.0002 to 20.0%) in matrix. Linearity of calibration curves in methanol was demonstrated with regression r2 ≥ 0.99. Within-batch precision (0.5 to 6.5%) and accuracy (91.4 to 108.0%) and between-batch precision (0.9 to 5.1%) and accuracy (91.5 to 107.5%) were demonstrated for quality control (QC) samples in methanol. Within-batch precision (0.2 to 3.6%) and accuracy (85.4 to 111.6%) and between-batch precision (1.4 to 6.1 %) and accuracy (90.2 to 110.3%) were also evaluated with a candidate cannabis certified reference material (CRM). Repeatability (1.5 to 12.4% RSD) and intermediate precision (2.2 to 12.8% RSD) were demonstrated via analysis of seven cannabis samples with HorRat values ranging from 0.3 to 3.1. The method provides enhanced detection limits coupled with a large quantitative range for 17 cannabinoids in plant material. It is suitable for a wide range of applications including routine analysis for delta-9-tetrahydrocannabinol (Δ9-THC), delta-9-tetrahydrocannabinolic acid (Δ9-THCA), cannabidiol (CBD), cannabidiolic acid (CBDA), and cannabinol (CBN) as well as more advanced interrogation of samples for both major and minor cannabinoids.


A SYBR Green I-based aptasensor for the label-free, fluorometric, and anti-interference detection of MeHg.
Methylmercury (MeHg+) is a common form of organic mercury that is substantially more toxic than inorganic mercury and is more likely to accumulate in organisms through biological enrichment. Therefore, developing a method to enable the specific and rapid detection of MeHg+ in seafood is important and remains challenging to accomplish. Herein, a rapid, label-free fluorescence detection method for MeHg+ determination was developed based on SYBR Green I. The detection system implemented "add and measure" detection mode can be completed in 10 min. Under optimal assay conditions, the detection platform showed a linear relationship with the concentration of MeHg+ within 1-50 nM (Y = 8.573x + 42.89, R2 = 0.9928), with a detection limit of 0.3218 nM. The results obtained for competitive substances, such as inorganic mercury ions and anions, show a high specificity of the method. In addition, this method successfully detected MeHg+ in seawater and marine products, with an accompanying spike recovery rate of 96.45-105.1%.


One-year trajectories of care and resource utilization for recipients of prolonged mechanical ventilation: a cohort study.
Growing numbers of critically ill patients receive prolonged mechanical ventilation. Little is known about the patterns of care as patients transition from acute care hospitals to postacute care facilities or about the associated resource utilization. To describe 1-year trajectories of care and resource utilization for patients receiving prolonged mechanical ventilation. 1-year prospective cohort study. 5 intensive care units at Duke University Medical Center, Durham, North Carolina. 126 patients receiving prolonged mechanical ventilation (defined as ventilation for >or=4 days with tracheostomy placement or ventilation for >or=21 days without tracheostomy), as well as their 126 surrogates and 54 intensive care unit physicians, enrolled consecutively over 1 year. Patients and surrogates were interviewed in the hospital, as well as 3 and 12 months after discharge, to determine patient survival, functional status, and facility type and duration of postdischarge care. Physicians were interviewed in the hospital to elicit prognoses. Institutional billing records were used to assign costs for acute care, outpatient care, and interfacility transportation. Medicare claims data were used to assign costs for postacute care. 103 (82%) hospital survivors had 457 separate transitions in postdischarge care location (median, 4 transitions [interquartile range, 3 to 5 transitions]), including 68 patients (67%) who were readmitted at least once. Patients spent an average of 74% (95% CI, 68% to 80%) of all days alive in a hospital or postacute care facility or receiving home health care. At 1 year, 11 patients (9%) had a good outcome (alive with no functional dependency), 33 (26%) had a fair outcome (alive with moderate dependency), and 82 (65%) had a poor outcome (either alive with complete functional dependency [4 patients; 21%] or dead [56 patients; 44%]). Patients with poor outcomes were older, had more comorbid conditions, and were more frequently discharged to a postacute care facility than patients with either fair or good outcomes (P < 0.05 for all). The mean cost per patient was $306,135 (SD, $285,467), and total cohort cost was $38.1 million, for an estimated $3.5 million per independently functioning survivor at 1 year. The results of this single-center study may not be applicable to other centers. Patients receiving prolonged mechanical ventilation have multiple transitions of care, resulting in substantial health care costs and persistent, profound disability. The optimism of surrogate decision makers should be balanced by discussions of these outcomes when considering a course of prolonged life support. None.


Endocrine responses to mother-infant separation in developing rats.
Separation of neonatal rat pups from the dam have been reported to elicit two endocrine responses in the pup: a fall in growth hormone secretion and a rise in corticosterone secretion. However, the temporal, ontogenetic, and behavioral determinants of these responses have not been compared. In the present study, we report that these two responses can be differentiated on each of these criteria. Growth hormone secretion falls rapidly immediately upon separation of pups from the dam, while robust rises in corticosterone secretion are delayed for many hours. In addition, growth hormone responses are observed earlier in ontogeny. Finally, active maternal behavior is required for normal growth hormone secretion in 10-day-old rat pups, while passive sensory stimuli associated with the dam can significantly reduce the corticosterone response to separation.