In vitro toxicity of fine and coarse particulate matter on the skin, ocular and lung microphysiological cell-culture systems.
Particulate matter (PM) has been associated with adverse effects on human health, causing allergies, skin and eye irritation and corrosion, respiratory tract irritation, headaches, bronchoconstriction, cardiopulmonary diseases such as asthma, chronic obstructive pulmonary disease (COPD), lung cancer, reproductive problems, premature deaths, and epigenetic changes that lead to a wide variety of cancers, among other health conditions. The air quality in the Medellín - Colombia presents fluctuations that oscillate between the maximum permissible levels established at the national level and by the WHO, which represents a latent risk to people's health. Although important efforts have been made to quantify the different levels of pollution and administrative measures have been established to mitigate air pollution, little research work has been done to establish the relationship between these levels of pollutants and the effects on biological systems. The objective of the present research was to make a morphological and chemical characterization of particulate matter (PM) captured with a commercial air filter and a electrospun nanofiber membrane and evaluate the cytotoxicity of the each PM extracts in monolayer and co-culture models which recreate microphysiological systems of lung, skin and cornea and propose the possible cellular interactions that lead the cytotoxic response of the chemical compounds found in particulate matter in cities. The morphology and elemental chemical characterization were done with scanning electron microscopy coupled with energy dispersive X-ray spectroscopy (SEM - EDS). For the polycyclic aromatic hydrocarbons detection was made with a chromatographic method accoupled to mass spectrometer. Finally, the cytotoxicity was made in monolayers of A549, HEK001, and SIRC cell lines and microphysiological systems consisting of two-cell layer construct to resemble the interaction between fibroblast and epithelial cells that comprises naturally the corneal, skin and lung tissue. We performed three different cocultures models with BALB/3T3 clone A31 as a feeder layer, using porous Transwell® inserts in the in-contact and non-contact way. Monolayer and co-culture models were exposed to coarse and fine PM (1, 2, and 5 mg/mL) and the cell viability was evaluated at 24 h using an MTT assay. The electrospun nanofibers membranes demonstrates higher efficiency to capture PM with different sizes and high concentration of polycyclic aromatic hydrocarbons, heavy metals, and other chemical compounds responsible of many human diseases. Cytotoxic effects of MP were observed in all models at higher concentration; however, models exposed to fine PM exhibited a significant reduction in cell viability compared to those exposed to coarse PM. In addition, multilayer models are more resistant to PM exposure than monolayer models. Furthermore, the study indicated that, depending on the seeding strategy, different results might be observed: the non-contact model showed higher resistance to PM exposure than in-contact for SIRC and HEK001, but A549 monolayers showed the highest viability response. This study demonstrates the usefulness of applying co-culture models to assess environmental pollutant toxicity, in addition to being a potential alternative method to animal testing for risk assessment.


Respiratory motoneuronal activity is altered by injections of picomoles of glutamate into cat brain stem.
The local neural circuitry underlying the control of breathing was studied by injecting nanoliter volumes of excitatory amino acids into discrete regions of cat brain stem. Experiments were performed on chloralose-urethane anesthetized, vagotomized, paralyzed, and artificially ventilated cats. Phrenic, intercostal, and recurrent laryngeal nerve discharges were recorded. Multibarrel pipettes were used for recording and pressure ejection of drugs or a dye for marking recording and ejection sites. Ejected volumes were directly monitored for every injection. Injections, proximal to neurons discharging with a respiratory periodicity, of as little of 200 fmol of L-glutamate in 200 pl of saline elicited marked, site-specific increases or decreases in respiratory motoneuronal discharge. N-Methyl-D-aspartic acid and homocysteic acid elicited similar site-specific alterations in respiratory motor output, although some details of the response could differ qualitatively. Responses to all the excitatory agents used were attenuated by concurrent injection of kynurenic acid, DL-2-amino-4-phosphonobutyric acid, or glutamic acid diethyl ester. There was no change in spontaneous phrenic nerve discharge in response to injections of equivalent or larger volumes of saline or lidocaine. These results indicate a heterogeneity in the spatial organization of the brain-stem neural circuitry underlying respiratory control, which has not been described previously. This injection technique may provide a mechanism for probing the neural circuitry underlying other behaviors.


Collective animal behavior from Bayesian estimation and probability matching.
Animals living in groups make movement decisions that depend, among other factors, on social interactions with other group members. Our present understanding of social rules in animal collectives is mainly based on empirical fits to observations, with less emphasis in obtaining first-principles approaches that allow their derivation. Here we show that patterns of collective decisions can be derived from the basic ability of animals to make probabilistic estimations in the presence of uncertainty. We build a decision-making model with two stages: Bayesian estimation and probabilistic matching. In the first stage, each animal makes a Bayesian estimation of which behavior is best to perform taking into account personal information about the environment and social information collected by observing the behaviors of other animals. In the probability matching stage, each animal chooses a behavior with a probability equal to the Bayesian-estimated probability that this behavior is the most appropriate one. This model derives very simple rules of interaction in animal collectives that depend only on two types of reliability parameters, one that each animal assigns to the other animals and another given by the quality of the non-social information. We test our model by obtaining theoretically a rich set of observed collective patterns of decisions in three-spined sticklebacks, Gasterosteus aculeatus, a shoaling fish species. The quantitative link shown between probabilistic estimation and collective rules of behavior allows a better contact with other fields such as foraging, mate selection, neurobiology and psychology, and gives predictions for experiments directly testing the relationship between estimation and collective behavior.


Development of total artificial heart with economical and durability advantages.
To develop a total artificial heart (TAH) pump system, we created a design paying particular attention to durability and cost. We adopted a pneumatically driven sac type artificial heart, where the configuration of the sac was decided according to the methodology of flow visualization. Its configuration is almost round to achieve as little stagnation as possible and a low turbulent flow. The main body of the sac was made using polyvinyl chloride (PVC) paste. The paste was poured into an external mold, and heated in a hot air drying oven. Coating was performed using polyurethane. The basic performance of this pump system was tested using a model circulation circuit, and a fitting study through acute animal experiment, using a healthy adult goat, was carried out. As for the TAH produced experimentally, a pump output exceeding 5.0 l/min in the model circulation circuit was provided. Implantation in the internal pleural cavity of a healthy adult goat, 55 kg in weight, proved possible and quite easy in comparison. It is thought that a more refined design in the connector part is desirable. Furthermore, a chronic experiment with the TAH will be carried out, and examination will need to be repeated in the future.


Rapamycin suppresses microglial activation and reduces the development of neuropathic pain after spinal cord injury.
Rapamycin is an inhibitor of the mammalian target of rapamycin (mTOR) signaling pathway, plays an important role in multiple cellular functions. Our previous study showed rapamycin treatment in acute phase reduced the neural tissue damage and locomotor impairment after spinal cord injury (SCI). However, there has been no study to investigate the therapeutic effect of rapamycin on neuropathic pain after SCI. In this study, we examined whether rapamycin reduces neuropathic pain following SCI in mice. We used a mouse model of thoracic spinal cord contusion injury, and divided the mice into the rapamycin-treated and the vehicle-treated groups. The rapamycin-treated mice were intraperitoneally injected with rapamycin (1 mg/kg) 4 h after SCI. The rapamycin treatment suppressed phosphorylated-p70S6K in the injured spinal cord that indicated inhibition of mTOR. The rapamycin treatment significantly improved not only locomotor function, but also mechanical and thermal hypersensitivity in the hindpaws after SCI. In an immunohistochemical analysis, Iba-1-stained microglia in the lumbar spinal cord was significantly decreased in the rapamycin-treated mice. In addition, the activity of p38 MAPK in the lumbar spinal cord was significantly attenuated by rapamycin treatment. Furthermore, phosphorylated-p38 MAPK-positive microglia was relatively decreased in the rapamycin-treated mice. These results indicated rapamycin administration in acute phase to reduce secondary neural tissue damage can contribute to the suppression of the microglial activation in the lumbar spinal cord and attenuate the development of neuropathic pain after SCI. The present study first demonstrated that rapamycin has significant therapeutic potential to reduce the development of neuropathic pain following SCI. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:93-103, 2017.


Growth and bioenergetics of alkaliphilic Bacillus firmus OF4 in continuous culture at high pH.
The effect of external pH on growth of alkaliphilic Bacillus firmus OF4 was studied in steady-state, pH-controlled cultures at various pH values. Generation times of 54 and 38 min were observed at external pH values of 7.5 and 10.6, respectively. At more alkaline pH values, generation times increased, reaching 690 min at pH 11.4; this was approximately the upper limit of pH for growth with doubling times below 12 h. Decreasing growth rates above pH 11 correlated with an apparent decrease in the ability to tightly regulate cytoplasmic pH and with the appearance of chains of cells. Whereas the cytoplasmic pH was maintained at pH 8.3 or below up to external pH values of 10.8, there was an increase up to pH 8.9 and 9.6 as the growth pH was increased to 11.2 and 11.4, respectively. Both the transmembrane electrical potential and the phosphorylation potential (delta Gp) generally increased over the total pH range, except for a modest fall-off in the delta Gp at pH 11.4. The capacity for pH homeostasis rather than that for oxidative phosphorylation first appeared to become limiting for growth at the high edge of the pH range. No cytoplasmic or membrane-associated organelles were observed at any growth pH, confirming earlier conclusions that structural sequestration of oxidative phosphorylation was not used to resolve the discordance between the total electrochemical proton gradient (delta p) and the delta Gp as the external pH is raised. Were a strictly bulk chemiosmotic coupling mechanism to account for oxidative phosphorylation over the entire range, the deltaGp/deltap ration (which would equal the H+/ATP ratio) would rise from about 3 at pH 7.5 to 13 at pH 11.2, dropping to 7 at pH 11.4 only because of the rise in cytoplasmic pH relative to other parameters. Moreover, the molar growth yields on malate were higher at pH 10.5 than at pH 7.5, indicating greater rather than lesser efficiency in the use of substrate at the more alkaline pH.


Whole-body [18F]fluoro-2-deoxyglucose positron emission tomography scan staging prior to planned radical hysterectomy and pelvic lymphadenectomy.
The aim of the study was to determine the effectiveness of whole-body [18F]fluoro-2-deoxyglucose positron emission tomography (FDG PET) imaging in properly selecting candidates for radical hysterectomy who are at low risk for subsequent chemoradiation. Retrospective study of 14 women undergoing planned radical hysterectomy and pelvic lymphadenectomy with clinically localized cervical cancer and either negative or inconclusive metastatic nodal disease by PET scan was performed. Pelvic lymph nodes were clearly negative by FDG PET scan in 12 of the 14 women. Two women had focal FDG uptake suspicious, although not definitive, for nodal metastasis, and pelvic nodes were positive at surgery in both. Computed tomography (CT) scan failed to detect nodal disease in either woman. Neither PET nor CT was effective at detecting parametrial disease, and both also failed to detect the primary tumor in some cases. Women with FDG PET scans that are clearly negative for nodal disease are good candidates for radical hysterectomy and are at low risk for subsequent chemoradiation.


[Age-related characteristics of testosterone 5-alpha-reductase activity in various rat organs].
A study was made of 5 alpha-reductase activity, transformation of 3H-testosterone to 3H-dihydrotestosterone in the liver, kidneys prostate an in the skin of the scrotum, as well as of the level of plasma testosterone in male Wistar rats at varying age periods of postnatal ontogenesis. The data obtained indicate that by the moment of birth the androgen-sensitive organs of male rats include functionally active 5 alpha-reductase. During body development, periods of the enzyme activation were discovered. It was shown that by senility androgen metabolism in the organs under study decelerates whereas, 5 alpha-dihydrotestosterone tends towards accumulation by the cells.


The role of triterpenoids in the topical anti-inflammatory activity of Calendula officinalis flowers.
By means of a bioassay-oriented fractionation of the CO2 extract of Calendula flowers, the triterpenoids are shown as the most important anti-inflammatory principles of the drug. Among them, the faradiol monoester appears to be the most relevant principle for the activity of the drug, due to its quantitative prevalence. The unesterified faradiol, not present in the extract, is the most active of the tested compounds and equals indomethacin in activity, whereas the monools psi-taraxasterol, lupeol, taraxasterol, and beta-amyrin are less active than the free diol. The anti-inflammatory activity of different CO2 extracts is proportional to their content of faradiol monoester, which can be taken as a suitable parameter for the quality control of Calendula preparations.


Hypoblastic tissue and fibroblast growth factor induce blood tissue (haemoglobin) in the early chick embryo.
We have investigated the temporal and the causal basis of blood tissue specification in the chick embryo. Earlier workers have shown that the prospective blood-forming area is specified in a horseshoe-shaped area at the posterior side of the embryo. We found that cultured explants from the posterior marginal zone at stages XI to XIII (consisting of the posterior marginal zone and part of Koller's sickle) have a high propensity to form haemoglobin (Hb), which could be inhibited at stage XI by adding antibody against basic fibroblast growth factor (bFGF) to the neutral culture medium; this treatment had no effect from stage XII onwards. The same result was found when whole embryos were cultured with an antiserum raised against bFGF, or with heparin. In another series of experiments, we found that cultured pieces from the inner-core of stage XIII epiblasts (with or without hypoblast tissue) were able to form Hb, whereas inner-core pieces from the pre-hypoblast stages, namely stages X and XI, did not form Hb. The capacity to form Hb, however, could be conferred upon the inner-core pieces from stage X epiblasts if bFGF at a concentration of 75-150 ng/ml was added to the culture medium. Furthermore, and most pertinently, the capacity to form Hb could be conferred on stage X inner-core pieces when they were co-cultured with hypoblast from a stage XIII embryo in a sandwich explant. Thus the inductive role of the hypoblast appears to be mediated via bFGF.(ABSTRACT TRUNCATED AT 250 WORDS)


Population genetic statistics from rhesus macaques (Macaca mulatta) in three different housing configurations at the California National Primate Research Center.
This study analyzed the genetic composition of 3382 genetically characterized and pedigreed animals currently maintained under 3 different housing configurations at the California National Primate Research Center, including the indoor colony, outdoor 'corn cribs,' and half-acre field cages. Summary statistics based on 15 short tandem repeats strongly suggest significant effects of genetic drift, including the loss of allele diversity, among the enclosures within the housing facilities even though gene flow among the different housing units is actively promoted by colony management. Management methods of selectively harvesting female macaques to prevent overrepresentation of one or only a few matrilines and cross-fostering 1-wk-old infants among breeding cages and corn cribs have been insufficient to prevent genetic subdivisions among the cages and corn cribs and to evenly distribute genetic diversity throughout the colony. In addition to promoting several colony management strategies recommended herein to effectively curb inbreeding and genetic differentiation, current attempts of infant cross-fostering and minimizing matriline fragmentation should be expanded. The inclusion of inbred or highly genetically homogeneous animals with diminished allele diversity in linkage and association studies will likely compromise the potential for identifying allele-disease associations, whereas the inclusion of macaques from different geographic origins or their hybrids (or both) in experimental research confounds interpretations of phenotypic differences, due to inflation of the genetic contribution to phenotypic variance.


Association of serum cytokines with oral HPV clearance.
Initial studies suggest higher serum levels of some pro-inflammatory cytokines may be associated with decreased cervical human papillomavirus (HPV) clearance. However, the relationship of cytokines with oral HPV clearance has not been explored. From 2010 to 2014, oral rinse and serum samples were collected semi-annually from 1601 adults. Oral rinse samples were tested for HPV DNA using PCR. Based on oral HPV results, 931 serum samples were selected for cytokine evaluation to include a roughly equal number of prevalent (n=307), incident (n=313), and no oral HPV infections (n=311). Electrochemiluminescence multiplex assays were used to determine the concentrations of IL-6, IL-8, TNF-α, IFN-γ, IL-1β, IL-2, IL-4, IL-10, IL-12 and IL-13. The relationship between serum cytokine concentrations (categorized into quartiles) and oral HPV clearance was evaluated with Wei-Lin-Weissfeld regression models, adjusting for HPV infection type (prevalent vs. incident), age, HIV status, and CD4 T cell count. Higher TNF-α concentration was associated with decreased clearance in men (highest vs. lowest quartile, adjusted hazard ratio [aHR]=0.52, 95% CI=0.34-0.79) and women (aHR=0.76, 95% confidence interval [CI]=0.55-1.04), with stronger associations in men than women (p-interaction=0.049). Higher IL-2 concentration was associated with reduced clearance in men (aHR=0.69, 95% CI=0.50-0.95), but not women (p-interaction=0.058). Results were similar within CD4 T cell strata (CD4⩾500 or CD4<500 cells/μl) among HIV-infected participants. No other cytokines were associated with clearance. High serum TNF-α is associated with reduced clearance of oral HPV infection.


Photoactivated Osmium Arene Anticancer Complexes.
Half-sandwich Os-arene complexes exhibit promising anticancer activity, but their photochemistry has hardly been explored. To exploit the photocytotoxicity and photochemistry of Os-arenes, O,O-chelated complexes [Os(η6-p-cymene)(Curc)Cl] (OsCUR-1, Curc = curcumin) and [Os(η6-biphenyl)(Curc)Cl] (OsCUR-2), and N,N-chelated complexes [Os(η6-biphenyl)(dpq)I]PF6 (OsDPQ-2, dpq = pyrazino[2,3-f][1,10]phenanthroline) and [Os(η6-biphenyl)(bpy)I]PF6 (OsBPY-2, bpy = 2,2'-bipyridine), have been investigated. The Os-arene curcumin complexes showed remarkable photocytotoxicity toward a range of cancer cell lines (blue light IC50: 2.6-5.8 μM, photocytotoxicity index PI = 23-34), especially toward cisplatin-resistant cancer cells, but were nontoxic to normal cells. They localized mainly in mitochondria in the dark but translocated to the nucleus upon photoirradiation, generating DNA and mitochondrial damage, which might contribute toward overcoming cisplatin resistance. Mitochondrial damage, apoptosis, ROS generation, DNA damage, angiogenesis inhibition, and colony formation were observed when A549 lung cancer cells were treated with OsCUR-2. The photochemistry of these Os-arene complexes was investigated by a combination of NMR, HPLC-MS, high energy resolution fluorescence detected (HERFD), X-ray adsorption near edge structure (XANES) spectroscopy, total fluorescence yield (TFY) XANES spectra, and theoretical computation. Selective photodissociation of the arene ligand and oxidation of Os(II) to Os(III) occurred under blue light or UVA excitation. This new approach to the design of novel Os-arene complexes as phototherapeutic agents suggests that the novel curcumin complex OsCUR-2, in particular, is a potential candidate for further development as a photosensitizer for anticancer photoactivated chemotherapy (PACT).


A fast neural network approach to predict lung tumor motion during respiration for radiation therapy applications.
During radiotherapy treatment for thoracic and abdomen cancers, for example, lung cancers, respiratory motion moves the target tumor and thus badly affects the accuracy of radiation dose delivery into the target. A real-time image-guided technique can be used to monitor such lung tumor motion for accurate dose delivery, but the system latency up to several hundred milliseconds for repositioning the radiation beam also affects the accuracy. In order to compensate the latency, neural network prediction technique with real-time retraining can be used. We have investigated real-time prediction of 3D time series of lung tumor motion on a classical linear model, perceptron model, and on a class of higher-order neural network model that has more attractive attributes regarding its optimization convergence and computational efficiency. The implemented static feed-forward neural architectures are compared when using gradient descent adaptation and primarily the Levenberg-Marquardt batch algorithm as the ones of the most common and most comprehensible learning algorithms. The proposed technique resulted in fast real-time retraining, so the total computational time on a PC platform was equal to or even less than the real treatment time. For one-second prediction horizon, the proposed techniques achieved accuracy less than one millimeter of 3D mean absolute error in one hundred seconds of total treatment time.


A Bayesian hierarchical model for demand curve analysis.
Drug self-administration experiments are a frequently used approach to assess the abuse liability and reinforcing property of a compound. It has been used to assess the abuse liabilities of various substances such as psychomotor stimulants and hallucinogens, food, nicotine, and alcohol. The demand curve generated from a self-administration study describes how demand of a drug or non-drug reinforcer varies as a function of price. With the approval of the 2009 Family Smoking Prevention and Tobacco Control Act, demand curve analysis provides crucial evidence to inform the US Food and Drug Administration's policy on tobacco regulation because it produces several important quantitative measurements to assess the reinforcing strength of nicotine. The conventional approach popularly used to analyze the demand curve data is individual-specific non-linear least square regression. The non-linear least square approach sets out to minimize the residual sum of squares for each subject in the dataset; however, this one-subject-at-a-time approach does not allow for the estimation of between- and within-subject variability in a unified model framework. In this paper, we review the existing approaches to analyze the demand curve data, non-linear least square regression, and the mixed effects regression and propose a new Bayesian hierarchical model. We conduct simulation analyses to compare the performance of these three approaches and illustrate the proposed approaches in a case study of nicotine self-administration in rats. We present simulation results and discuss the benefits of using the proposed approaches.


The Khoekhoegowab Personality Inventory: The Comparative Validity of a Locally Derived Measure of Traits.
Objective: This study explores a personality inventory derived from the results of an indigenous lexical study of personality. From the 272 most commonly used personality descriptors in Khoekhoegowab, the most-spoken of extant Khoesan click languages of southern Africa, an 11-factor model of personality-trait structure was identified. Here, the Khoekhoegowab Personality Inventory (KPI) was created based on those results. Its psychometric properties, the convergent and divergent validity of its scales, and its incremental validity over Big Five and Six traits for predicting physical and mental health, religious practice and attitudes, and income are reported. Methods: Two to five key terms were selected for each of 10 KPI scales: Temperance, Prosocial Diligence, Gossip, Honesty/Morality, Temper, Implacability, Humility, Vanity, Resiliency vs. Agitation, and Courage vs. Fear. These 38 total items were administered to a large sample of adult speakers of Khoekhoegowab in Namibia (N = 632), together with five imported inventories translated into Khoekhoegowab: the 30-item Questionnaire Big Six (QB6), General Self-Reported Health, the Cascades Mental Health Assessment, the Satisfaction with Life Scale, the Duke Religion Index. The properties and intercorrelations of KPI subscales are explored, and their predictive ability for the other variables is compared to that of the QB6. Results: Due to the small number of items on each scale, poor internal consistency was anticipated, but the KPI scales' properties were somewhat better than those of the QB6. R-square change by the inventories as a whole, after accounting for age and gender, indicted that the KPI scales explained more variance than the QB6 scales in almost all criterion variables. Replication of established associations for Big Six traits was mixed: associations were largely as expected for Resiliency, Conscientiousness, and Honesty, but less so for Agreeableness and Extraversion. Conclusions: The KPI had some advantages over the QB6 in predicting physical and mental health. In particular, the four items of Resiliency vs. Agitation predicted lower scores on all physical and mental problem scales. Given psychological-care needs in Namibia, this might be used as a non-intrusive screener. Measurement challenges common to both surveys are discussed, possible solutions, and the utility of higher-order structures are discussed.


GLOBIN-5-dependent O2 responses are regulated by PDL-1/PrBP that targets prenylated soluble guanylate cyclases to dendritic endings.
Aerobic animals constantly monitor and adapt to changes in O2 levels. The molecular mechanisms involved in sensing O2 are, however, incompletely understood. Previous studies showed that a hexacoordinated globin called GLB-5 tunes the dynamic range of O2-sensing neurons in natural C. elegans isolates, but is defective in the N2 lab reference strain (McGrath et al., 2009; Persson et al., 2009). GLB-5 enables a sharp behavioral switch when O2 changes between 21 and 17%. Here, we show that GLB-5 also confers rapid behavioral and cellular recovery from exposure to hypoxia. Hypoxia reconfigures O2-evoked Ca(2+) responses in the URX O2 sensors, and GLB-5 enables rapid recovery of these responses upon re-oxygenation. Forward genetic screens indicate that GLB-5's effects on O2 sensing require PDL-1, the C. elegans ortholog of mammalian PrBP/PDE6δ protein. In mammals, PDE6δ regulates the traffic and activity of prenylated proteins (Zhang et al., 2004; Norton et al., 2005). PDL-1 promotes localization of GCY-33 and GCY-35, atypical soluble guanylate cyclases that act as O2 sensors, to the dendritic endings of URX and BAG neurons, where they colocalize with GLB-5. Both GCY-33 and GCY-35 are predicted to be prenylated. Dendritic localization is not essential for GCY-35 to function as an O2 sensor, but disrupting pdl-1 alters the URX neuron's O2 response properties. Functional GLB-5 can restore dendritic localization of GCY-33 in pdl-1 mutants, suggesting GCY-33 and GLB-5 are in a complex. Our data suggest GLB-5 and the soluble guanylate cyclases operate in close proximity to sculpt O2 responses.


Massive pulmonary embolism: embolectomy or thrombolysis?
The purpose of this paper is to evaluate the indications for surgical or thrombolytic treatment of massive pulmonary embolism. Analysis of the outcome of the disease depending on the indices of lung vascular damage volume and hemodynamics was performed. One hundred and thirty five patients with thromboembolism of truncus or main branches of the pulmonary artery were examined. Right cardiac catheterization, angiopulmonography and Tc-perfusion lung scanning were carried out. Predictive criteria of the lethal outcome (with 86% probability) were defined in patients without surgical treatment. These criteria included extensive volume of lung vascular damage (Miller angiographic index greater than or equal to 27, scannographic perfusion deficit greater than or equal to 60%) and significant hemodynamic disturbances (refractory systemic arterial hypotension or severe pulmonary arterial hypertension: right ventricular systolic pressure greater than or equal to 60 mmHg, right ventricular end-diastolic pressure greater than or equal to 15 mmHg, mean right ventricular pressure greater than or equal to 25 mmHg, and mean pulmonary arterial pressure greater than or equal to 35 mmHg). With these indices available these are indications for pulmonary artery embolectomy. With lower indices, but close to those estimated in the study, thrombolytic therapy is indicated.


The response of PKD1L3/PKD2L1 to acid stimuli is inhibited by capsaicin and its pungent analogs.
Polycystic kidney disease (PKD) 2L1 protein is a member of the transient receptor potential (TRP) ion channel family. In circumvallate and foliate papillae, PKD2L1 is coexpressed with PKD1L3. PKD2L1 and PKD1L3 interact through their transmembrane domain and the resulting heteromer PKD1L3/PKD2L1 owns a unique channel property called 'off-responses' to acid stimulation, although PKD2L1 does not own this property by itself. To define the pharmacological properties of the PKD1L3/PKD2L1 channel, we developed a new method to effectively evaluate channel activity using human embryonic kidney 293T cells in which the channel was heterologously expressed. This method was applied to screen substances that potentially regulate it. We found that capsaicin and its analogs, which are TRPV1 agonists, inhibited the response to acid stimuli and that the capsaicin inhibition was reversible with an IC(50) of 32.5 μm. Capsaicin and its analogs are thus useful tools for physiological analysis of PKD1L3/PKD2L1 function.


c-Jun N-terminal kinase pathway activation in human and experimental cerebral contusion.
The c-Jun N-terminal kinase (JNK) pathway is involved in cell stress and apoptosis. We tested the hypothesis that this pathway plays a role in traumatic brain injury (TBI) by assessing JNK activation in human brain tissues and in brains of mice subjected to controlled cortical impact brain injury. We also assessed the effects of specific inhibition of the JNK pathway by the cell-permeable JNK inhibitor peptide, D-JNKI1, on neurobehavioral function and posttraumatic cell loss in mice. The inhibitor was administered intraperitoneally 10 minutes after injury. The JNK pathway showed robust activation both in human contusion specimens and in injured cortex and hippocampi of TBI-injured mice, 1, 4, and 48 hours after injury. D-JNKI1 treatment significantly improved motor performance at 48 hours and 7 days after injury and reduced the contusion volume compared with saline treatment; the numbers of terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling-positive cells were significantly decreased in the hippocampi of injured mice 48 hours after treatment. Thus, because the JNK pathway is activated after human and experimental TBI and the inhibitor peptide D-JNKI1 affords significant neuroprotection and amelioration of neurobehavioral deficits after experimental TBI, therapeutic targeting of the JNK activation pathway may hold promise for future clinical applications.


A taxonomy and framework for identifying and developing actionable statements in guidelines suggests avoiding informal recommendations.
To propose a taxonomy and framework that identifies and presents actionable statements in guidelines. We took an iterative approach reviewing case studies of guidelines produced by the World Health Organization and the American Society of Hematology to develop an initial conceptual framework. We then tested it using randomly selected recommendations from published guidelines addressing COVID-19 from different organizations, evaluated its results, and refined it before retesting. The urgency and availability of evidence for development of these recommendations varied. We consulted with experts in research methodology and guideline developers to improve the final framework. The resulting taxonomy and framework distinguishes five types of actional statements: formal recommendations; research recommendations; good practice statements; implementation considerations, tools and tips; and informal recommendations. These statements should respond to a priori established criteria and require a clear structure and recognizable presentation in a guideline. Most importantly, this framework identifies informal recommendations that differ from formal recommendations by how they consider evidence and in their development process. The identification, standardization and explicit labelling of actionable statements according to the framework may support guideline developers to create actionable statements with clear intent, avoid informal recommendations and improve their understanding and implementation by users.


Protein identification with a single accurate mass of a cysteine-containing peptide and constrained database searching.
A method for rapid and unambiguous identification of proteins by sequence database searching using the accurate mass of a single peptide and specific sequence constraints is described. Peptide masses were measured using electrospray ionization-Fourier transform ion cyclotron resonance mass spectrometry to an accuracy of 1 ppm. The presence of a cysteine residue within a peptide sequence was used as a database searching constraint to reduce the number of potential database hits. Cysteine-containing peptides were detected within a mixture of peptides by incorporating chlorine into a general alkylating reagent specific for cysteine residues. Secondary search constraints included the specificity of the protease used for protein digestion and the molecular mass of the protein estimated by gel electrophoresis. The natural isotopic distribution of chlorine encoded the cysteine-containing peptide with a distinctive isotopic pattern that allowed automatic screening of mass spectra. The method is demonstrated for a peptide standard and unknown proteins from a yeast lysate using all 6118 possible yeast open reading frames as a database. As judged by calculation of codon bias, low-abundance proteins were identified from the yeast lysate using this new method but not by traditional methods such as tandem mass spectrometry via data-dependent acquisition or mass mapping.


Proliferative activity of intrinsic cell populations in the normal human kidney.
The proliferative activity of various normal human renal cell populations is unknown. Recently, antibodies to cell proliferation-associated nuclear proteins, such as proliferating cell nuclear antigen (PCNA) and KI-67, which are applicable to archival paraffin sections, became available. With antibodies to PCNA and Ki-67 after microwave pretreatment of the paraffin sections, the proliferation indexes (ratio of positive nuclei with PCNA and Ki-67 antibodies/all nuclei counted x 100, i.e. percentage of positive cells) of 12 different intrinsic renal cell populations in 20 normal human kidneys have been determined. The following proliferation indexes (percentages of positive cells) were found with the PCNA and the Ki-67 antibodies, respectively: proximal tubular epithelium, 0.22, 0.24; thin limb of Henle, 0.29, 0.30; thick ascending limb of Henle, 0.32, 0.29; distal tubular epithelium (distal convoluted tubules and cortical collecting ducts, 0.33, 0.44; medullary collecting ducts, 0.32, 0.3; glomerular mesangial cells, 0.07, 0.12; glomerular visceral epithelial cells, 0.04, 0.08; glomerular parietal epithelial cells, 0.07, 0.1; glomerular capillary endothelium, 0.42, 0.47; peritubular capillary endothelial cells, 0.38, 0.43; endothelium of large intrarenal vessels (arteries and veins), 0.09, 0.12. Thus, normally capillary endothelium (glomerular and peritubular) appears to have the highest proliferation index in the human kidney by these techniques. These results indicate major variation in the proliferative activity of normal human renal cell populations, along with a significant correlation between PCNA and Ki-67 staining. Furthermore, this study provides normal values for the proliferative activity of different human renal cell populations.(ABSTRACT TRUNCATED AT 250 WORDS)


Typical intellectual engagement and cognition in old age.
Typical Intellectual Engagement (TIE) comprises the preference to engage in cognitively demanding activities and has been proposed as a potential explanatory variable of individual differences in cognitive abilities. Little is known, however, about the factorial structure of TIE, its relations to socio-demographic variables, and its influence on intellectual functioning in old age. In the present study, data of 364 adults (65-81 years) from the Zurich Longitudinal Study on Cognitive Aging (ZULU) were used to investigate the factorial structure of TIE and to examine the hypothesis that TIE is associated more strongly with crystallized intelligence than with fluid intelligence in old age. A measurement model of a second order factor based on a structure of four correlated first order factors (Reading, Problem Solving, Abstract Thinking, and Intellectual Curiosity) evinced an excellent fit. After controlling for age, sex, and formal education, TIE was more strongly associated with crystallized intelligence than with fluid intelligence, comparable to results in younger persons. More detailed analyses showed that this association is mostly defined via Reading and Intellectual Curiosity.


Lymphomas in patients with Sjogren's syndrome are marginal zone B-cell neoplasms, arise in diverse extranodal and nodal sites, and are not associated with viruses.
The occurrence of non-Hodgkin's lymphoma (NHL) is the most serious complication of Sjogren's syndrome (SS). We performed a study of 16 NHLs occurring in patients with an underlying SS. These lymphomas arose not only in salivary glands (7 cases) but also in other mucosal extranodal sites (the stomach [4 cases], the lung [3 cases], the skin [3 cases], the buccal mucosa [1 case], the thymus [1 case]) and in nodal sites (8 cases). Low-grade marginal zone lymphomas (MZL) were diagnosed in 12 of the 16 patients, 9 of mucosa-associated lymphoid tissues (MALT) type in mucosal sites and 3 exclusively nodal. The 4 other patients presented with a high-grade B-cell lymphoma that was probably a histological transformation of an underlying low-grade MZL at least in 3 of the cases involving skin, stomach, and parotid, respectively. A t(14;18) translocation was detected in 1 of 8 lymphomas tested. We detected serum anti-p53 antibodies in 2 of the 14 studied patients. p53 protein was detected in 1 of 11 lymphomas tested. LMP protein and Eber RNAs of Epstein-Barr virus (EBV) were not detected in the 16 NHL biopsies. Using polymerase chain reaction, EBV was never detected except in 1 of 4 parotid lymphomas. No human T-lymphotropic virus 1 or human herpes virus 8 DNAs were detected in NHL biopsies. None of the patients had hepatitis C virus infection found using serological methods. Chemotherapy was usually efficient. In conclusion, lymphomas occurring in patients with an underlying SS are in most cases MZL. These lymphomas are not associated with viruses known to be present in other types of lymphomas. Some of the translocations or mutations of oncogenes or antioncogenes described in other lymphomas are detected in SS-associated lymphomas.


Dissecting the effects of nitrate, sucrose and osmotic potential on Arabidopsis root and shoot system growth in laboratory assays.
Studying the specific effects of water and nutrients on plant development is difficult because changes in a single component can often trigger multiple response pathways. Such confounding issues are prevalent in commonly used laboratory assays. For example, increasing the nitrate concentration in growth media alters both nitrate availability and osmotic potential. In addition, it was recently shown that a change in the osmotic potential of media alters the plant's ability to take up other nutrients such as sucrose. It can also be difficult to identify the initial target tissue of a particular environmental cue because there are correlated changes in development of many organs. These growth changes may be coordinately regulated, or changes in development of one organ may trigger changes in development of another organ as a secondary effect. All these complexities make analyses of plant responses to environmental factors difficult to interpret. Here, we review the literature on the effects of nitrate, sucrose and water availability on root system growth and discuss the mechanisms underlying these effects. We then present experiments that examine the impact of nitrate, sucrose and water on root and shoot system growth in culture using an approach that holds all variables constant except the one under analysis. We found that while all three factors also alter root system size, changes in sucrose and osmotic potential also altered shoot system size. In contrast, we found that, when osmotic effects are controlled, nitrate specifically inhibits root system growth while having no effect on shoot system growth. This effectively decreases the root : shoot ratio. Alterations in root : shoot ratio have been widely observed in response to nitrogen starvation, where root growth is selectively increased, but the present results suggest that alterations in this ratio can be triggered across a wide spectrum of nitrate concentrations.


Promoter region sequence differences in the A and G gamma globin genes of Brazilian sickle cell anemia patients.
Fetal hemoglobin (HbF), encoded by the HBG2 and HBG1 genes, is the best-known genetic modulator of sickle cell anemia, varying dramatically in concentration in the blood of these patients. This variation is partially associated with polymorphisms located in the promoter region of the HBG2 and HBG1 genes. In order to explore known and unknown polymorphisms in these genes, the sequences of their promoter regions were screened in sickle cell anemia patients and correlated with both their HbF levels and their betaS-globin haplotypes. Additionally, the sequences were compared with genes from 2 healthy groups, a reference one (N = 104) and an Afro-descendant one (N = 98), to identify polymorphisms linked to the ethnic background.The reference group was composed by healthy individuals from the general population. Four polymorphisms were identified in the promoter region of HBG2 and 8 in the promoter region of HBG1 among the studied groups. Four novel single nucleotide polymorphisms (SNP) located at positions -324, -317, -309 and -307 were identified in the reference group. A deletion located between -396 and -391 in the HBG2 promoter region and the SNP -271 C-->T in the HBG1 promoter region were associated with the Central African Republic betaS-globin haplotype. In contrast, the -369 C-->G and 309 A-->G SNPs in the HBG2 promoter region were correlated to the Benin haplotype. The polymorphisms -396_-391 del HBG2, -369 SNP HBG2 and -271 SNP HBG1 correlated with HbF levels. Hence, we suggest an important role of HBG2 and HBG1 gene polymorphisms on the HbF synthesis.


Evaluation of nitric oxide synthase activity, nitric oxide, and homocysteine levels in patients with active Behcet's disease.
Behcet's disease (BD) is a chronic, progressive disorder that affects many systems of the body including the eye. The aim of this study was to examine the effects of nitric oxide synthase activity (NOS), nitric oxide (NO*), and homocysteine (Hcy) levels in patients with active BD. Included in this study were 18 male BD patients and 16 male healthy volunteers as controls. Erythrocyte NOS activity, (NO*) erythrocyte sedimentation rate, serum C-reactive protein, and plasma Hcy values in the patients with BD were significantly higher than those of the control group. Our results show that these parameters play a major role in the inflammatory reactions observed in BD.


Comparing closed quotient in children singers' voices as measured by high-speed-imaging, electroglottography, and inverse filtering.
The closed quotient, i.e., the ratio between the closed phase and the period, is commonly studied in voice research. However, the term may refer to measures derived from different methods, such as inverse filtering, electroglottography or high-speed digital imaging (HSDI). This investigation compares closed quotient data measured by these three methods in two boy singers. Each singer produced sustained tones on two different pitches and a glissando. Audio, electroglottographic signal (EGG), and HSDI were recorded simultaneously. The audio signal was inverse filtered by means of the decap program; the closed phase was defined as the flat minimum portion of the flow glottogram. Glottal area was automatically measured in the high speed images by the built-in camera software, and the closed phase was defined as the flat minimum portion of the area-signal. The EGG-signal was analyzed in four different ways using the matlab open quotient interface. The closed quotient data taken from the EGG were found to be considerably higher than those obtained from inverse filtering. Also, substantial differences were found between the closed quotient derived from HSDI and those derived from inverse filtering. The findings illustrate the importance of distinguishing between these quotients.


Hypoxia in Bone and Oxygen Releasing Biomaterials in Fracture Treatments Using Mesenchymal Stem Cell Therapy: A Review.
Bone fractures have a high degree of severity. This is usually a result of the physical trauma of diseases that affect bone tissues, such as osteoporosis. Due to its highly vascular nature, the bone is in a constant state of remodeling. Although those of younger ages possess bones with high regenerative potential, the impact of a disrupted vasculature can severely affect the recovery process and cause osteonecrosis. This is commonly seen in the neck of femur, scaphoid, and talus bone. In recent years, mesenchymal stem cell (MSC) therapy has been used to aid in the regeneration of afflicted bone. However, the cut-off in blood supply due to bone fractures can lead to hypoxia-induced changes in engrafted MSCs. Researchers have designed several oxygen-generating biomaterials and yielded varying degrees of success in enhancing tissue salvage and preserving cellular metabolism under ischemia. These can be utilized to further improve stem cell therapy for bone repair. In this review, we touch on the pathophysiology of these bone fractures and review the application of oxygen-generating biomaterials to further enhance MSC-mediated repair of fractures in the three aforementioned parts of the bone.


Janus-faced microglia: beneficial and detrimental consequences of microglial phagocytosis.
Microglia are the resident brain macrophages and they have been traditionally studied as orchestrators of the brain inflammatory response during infections and disease. In addition, microglia has a more benign, less explored role as the brain professional phagocytes. Phagocytosis is a term coined from the Greek to describe the receptor-mediated engulfment and degradation of dead cells and microbes. In addition, microglia phagocytoses brain-specific cargo, such as axonal and myelin debris in spinal cord injury or multiple sclerosis, amyloid-β deposits in Alzheimer's disease, and supernumerary synapses in postnatal development. Common mechanisms of recognition, engulfment, and degradation of the different types of cargo are assumed, but very little is known about the shared and specific molecules involved in the phagocytosis of each target by microglia. More importantly, the functional consequences of microglial phagocytosis remain largely unexplored. Overall, phagocytosis is considered a beneficial phenomenon, since it eliminates dead cells and induces an anti-inflammatory response. However, phagocytosis can also activate the respiratory burst, which produces toxic reactive oxygen species (ROS). Phagocytosis has been traditionally studied in pathological conditions, leading to the assumption that microglia have to be activated in order to become efficient phagocytes. Recent data, however, has shown that unchallenged microglia phagocytose apoptotic cells during development and in adult neurogenic niches, suggesting an overlooked role in brain remodeling throughout the normal lifespan. The present review will summarize the current state of the literature regarding the role of microglial phagocytosis in maintaining tissue homeostasis in health as in disease.


Changes in symptoms and interpersonal problems during the first 2 years of long-term psychoanalytic psychotherapy and psychoanalysis.
Longitudinal measurements can provide important information regarding variations in developmental trajectories of patients in long-term treatment. The present study investigated changes in general symptoms, depression, anxiety, and interpersonal problems during the first 2 years of long-term psychoanalytic psychotherapy (PP) and psychoanalysis (PA). It was expected that interpersonal problems would diminish more slowly compared to symptomatic dysfunction. An accelerated longitudinal design with five consecutive measurement points across two cohorts of patients was used. Changes on the Symptom Checklist-90-R (SCL-90-R), Beck Depression Inventory-II (BDI-II), State-Trait Anxiety Inventory (STAI), and Inventory of Interpersonal Problems-64 (IIP-64) were investigated during the first 2 years of long-term PP (n = 73) and PA (n = 40). Linear regression analysis was performed to model the different courses of improvement. After 2 years of treatment, patients in both groups still presented moderate to high levels of symptoms and interpersonal problems compared to non-clinical populations. As expected, interpersonal problems changed less rapidly. PP patients changed both with regard to symptomatic and interpersonal problems, whereas the only significant change in the PA group was on one of the symptomatic subscales. Slopes in the PA group and in PP group did not differ significantly from each other, except for the IIP-64 scale intrusive, with PP patients showing significantly more improvement than PA patients. The height of intake values of the outcome variables appeared to predict the speed of symptomatic recovery. Symptoms and interpersonal problems did not decrease notably within the first 2 years of psychoanalytic treatment. This is consistent with the idea that significant change takes time for patients with chronic mental disorders and personality pathology. In regular practice, it is advisable to monitor changes routinely in order to identify slow responders more quickly and change the treatment plan, if necessary.


Scleral necrosis in patients with posterior uveal melanomas evaluated by transcleral fine needle aspiration biopsy and treated by 125I plaque.
To evaluate the incidence, potential correlation with transcleral fine needle aspiration biopsy, and treatment of scleral necrosis in patients with posterior uveal melanomas treated by 125I plaque radiotherapy and assessed by transcleral fine needle aspiration biopsy. We per-formed a retrospective review of posterior uveal melanoma treated by 125I plaque radiotherapy at a single academic institution between July 2006 and July 2013. Consecutive patients diagnosed with a posterior uveal melanoma during the study period that had an anterior margin at or anterior to the equator who were evaluated by transcleral fine needle aspiration biopsy prior to 125I plaque radiotherapy were included. The main outcome measure was development of scleral necrosis, and the secondary outcome was treatment of this complication. Statistical analysis included computation of conventional descriptive statistics, cross-tabulation and chi-square tests of potential factors related to the development of scleral necrosis, and summarizing of treatment approaches and results. The incidence of treatment of scleral necrosis was calculated using the Kaplan-Meier method. During the 7-year study period, 87 posterior uveal melanomas were evaluated by transcleral fine needle aspiration biopsy and treated by 125I plaque radiotherapy. The median largest basal diameter of the tumor was 13.3 mm, and the median thickness was 6.8 mm. Eight patients (9.2%) developed scleral necrosis during follow-up. Thicker tumors (> 6.5 mm) were more likely to develop scleral necrosis (n=7) than thinner tumors (p=0.05). The median interval between 125I plaque radiotherapy and detection of scleral necrosis was 19.1 months. The overall cumulative probability of scleral necrosis was 6.2% at 6 months and 14.3% at 24 months, subsequently remaining stable. For thicker tumors, the probability of scleral necrosis was 23.5% at 45.4 months. Five patients were treated by scleral patch graft (62.5%) and three by observation (37.5%). One patient underwent enucleation after two failed scleral patch attempts and recurrent scleral necrosis. The mean follow-up period for patients with scleral necrosis was 34.5 months. Thicker posterior uveal melanomas are more likely to develop scleral necrosis after 125I plaque radiotherapy and transcleral fine needle aspiration biopsy. While observation is sufficient for managing limited scleral necrosis, scleral patch graft is a viable alternative for eye preservation in extensive scleral necrosis.


Nurse-led culturally relevant smoking reduction intervention among cardiovascular and respiratory patients in Pakistan.
Socioeconomically deprived populations are at greater risk for smoking-induced diseases and death, such as cancers, and cardiovascular and respiratory illnesses. The initiation of a nurse-led smoking cessation program in clinical practice is an effective method to enhance smoking cessation among cardiovascular and respiratory patients in Pakistan. This study aimed to evaluate the effectiveness of nurse-led smoking reduction intervention performed at out-patient clinics in Karachi, Pakistan. A single group pre-and post-test study was conducted on eligible patients (n = 83) with a typical profile of cardiovascular and respiratory diseases in a tertiary care hospital from December 2020 to July 2021. The multi-modal nurse-led intervention consisted of two face-to-face motivational counseling (30-min) and free nicotine replacement therapy with telephonic counseling and follow-ups offered to all eligible patients for 2 months. The nurse-led intervention resulted in a significant decrease in daily cigarette consumption in most of the patients (75.9%) by 50% and 16.9% of the patients reported smoking abstinence (quitter) at 1-week follow-up (p < .001), verified by a CO breath test. A nurse-managed smoking reduction intervention for even a short duration (2 months) in clinics is an effective approach in enhancing smoking abstinence and reduction among cardiovascular and respiratory patients.


A benchmark study of simulation methods for single-cell RNA sequencing data.
Single-cell RNA-seq (scRNA-seq) data simulation is critical for evaluating computational methods for analysing scRNA-seq data especially when ground truth is experimentally unattainable. The reliability of evaluation depends on the ability of simulation methods to capture properties of experimental data. However, while many scRNA-seq data simulation methods have been proposed, a systematic evaluation of these methods is lacking. We develop a comprehensive evaluation framework, SimBench, including a kernel density estimation measure to benchmark 12 simulation methods through 35 scRNA-seq experimental datasets. We evaluate the simulation methods on a panel of data properties, ability to maintain biological signals, scalability and applicability. Our benchmark uncovers performance differences among the methods and highlights the varying difficulties in simulating data characteristics. Furthermore, we identify several limitations including maintaining heterogeneity of distribution. These results, together with the framework and datasets made publicly available as R packages, will guide simulation methods selection and their future development.


Nurse-led peer support group: experiences of women with polycystic ovary syndrome.
This paper is a report of a study to explore the experiences of women with polycystic ovary syndrome attending a nurse-led support group. Polycystic ovary syndrome is a common chronic endocrine disorder associated with high levels of psychological distress. It has been argued that healthcare providers should regularly review the psychological health of women with polycystic ovary syndrome, and that nurses can help women to adjust to the condition by providing education and support. Little is known about the means of providing social support for women with the syndrome, or of any benefits for patients. Qualitative interviews were conducted with 13 female patients in 2006. The patients attended a support group at a public hospital in the United Kingdom. The data were analysed using deductive and inductive thematic analysis. The group provided both socio-emotional and informational social support. Participants reported that attending the group helped to reduce isolation, and provided an opportunity for social comparison and accessible and personally relevant information. Participants described the group as having had a major personal impact for them. They reported feeling empowered and direct positive effects on their self-management behaviours. Social support appears to be a key factor mediating the psychosocial impact in women with polycystic ovary syndrome. Healthcare providers should consider running such support groups or referring patients to them. Providing social support may alleviate distress and improve self-management.


Pituitary-gonadal and pituitary-thyroid axis hormone concentrations before and during a hypoglycemic clamp after sleep deprivation in healthy men.
Total sleep deprivation (TSD) exerts strong modulatory effects on the secretory activity of endocrine systems that might be related to TSD-induced challenges of cerebral glucose metabolism. Here, we investigate whether TSD affects the course of male pituitary-gonadal and pituitary-thyroid axis related hormones during a subsequent 240-min hypoglycemic clamp. Ten healthy men were tested on 2 different conditions, TSD and 7-hour regular sleep. Circulating concentrations of total testosterone, prolactin (PRL), thyroid stimulating hormone (TSH), free triiodothyronine (fT3), and free thyroxin (fT4) were measured during baseline and a subsequent hypoglycemic clamp taking place in the morning. Basal, i.e. at 07:00 am measured, concentrations of total testosterone (P = 0.05) and PRL (P<0.01) were lower while the values of TSH (P = 0.02), fT3 (P = 0.08), and fT4 (P = 0.04) were higher after TSD as compared to regular sleep. During the subsequent hypoglycemic clamp (all measurements from baseline to the end of the clamp analyzed) total testosterone concentrations in the regular sleep (P<0.01) but not in the TSD condition (P = 0.61) decreased, while PRL levels increased (P = 0.05) irrespectively of the experimental condition (P = 0.31). TSH concentrations decreased during hypoglycemia (P<0.01), with this decrease being more pronounced after TSD (P = 0.04). However, at the end of the hypoglycemic clamp concentrations all of the above mentioned hormones did not differ between the two sleep conditions. Our data indicate a profound influence of TSD on male pituitary-gonadal and pituitary-thyroid axis hormones characterized by reduced basal testosterone and PRL levels and increased TSH levels. However, since concentrations of these hormones measured at the end of the 240-min hypoglycemic clamp were not affected by TSD it can be speculated that the influence of TSD on the two endocrine axes is rather short lived or does not interact in an additive manner with their responses to hypoglycemia.


Immunocytochemical localization of intracellular acidic compartments: rat proximal nephron.
There is evidence that components of the endosomal-lysosomal system of most cells have an acidic interior. The weak base, N-(3-[(2,4-dinitrophenyl)amino]propyl)-N-(3-aminopropyl)methylamine dihydrochloride (DAMP) has been shown to accumulate in acidic compartments in cultured cells and in isolated perfused proximal tubules. We infused DAMP intravascularly in vivo and used colloidal gold immunocytochemistry to identify acidic compartments in cells of the rat glomerulus and proximal tubule. Sprague-Dawley rats were infused intra-arterially with DAMP. The kidneys were fixed by intravascular perfusion with 1% glutaraldehyde and embedded in Lowicryl K4M. Sections were exposed to a mouse anti-dinitrophenol (DNP) monoclonal antibody that cross-reacts with DAMP, followed by gold-conjugated goat anti-mouse immunoglobulin G. Transmission electron microscopy revealed positive labeling of the lysosome-like structures of the various cells of the glomerulus and lysosomes, endosomes, and numerous endocytic vesicles of all segments of the proximal tubule. We conclude that 1) DAMP can be used in vivo to identify acidic compartments in the kidney and 2) lysosomes, endosomes, and many endocytic vesicles of the rat proximal tubule as well as lysosome-like structures in cells of the glomerulus have an acidic interior.


Eradication of Salmonella and Arizona species from turtle hatchlings produced from eggs treated on commercial turtle farms.
On commercial turtle farms more than 40% of the hatchlings excrete detectable levels of Salmonella and Arizona spp. when hatched from nonsanitized eggs incubated in sawdust or dirt-filled chambers. Over a 3-year period on 10 farms, more than 10(6) turtle eggs were treated in an attempt to hatch Salmonella-free turtles. Eggs were sanitized in disinfectant, treated by temperature- or pressure-differential dip methods in solutions containing 500 micrograms or more of gentamicin sulfate per ml, and hatched in sanitized plastic chambers free of bedding material. The Salmonella and Arizona spp. infection levels for turtles produced from treated eggs were 0 and 1.12% for years 1 and 2, respectively, whereas infection levels for hatchlings produced from nontreated eggs during these periods were 47 and 44%, respectively. During year 3, dip solutions were filtered daily, treated at 100 degrees C for 15 min on a weekly basis to free the solution of microbial contaminants and egg protein, charged with gentamicin after 10,000 to 20,000 eggs had been treated to maintain antimicrobial activity at 500 micrograms/ml or more, and maintained at pH 6.0 to preserve optimal antimicrobial activity. The implementation of these measures in year 3 resulted in an infection level of 0.15% when the tissues of 3 of 1,959 hatchlings tested were positive for Salmonella and Arizona spp., whereas the tissues of 66 (49.0%) of 135 hatchlings produced from nontreated eggs were positive.


Anti-IgE autoantibodies in asthma: a diagnostic artefact or an explanation for non-allergic asthma?
Autoantibodies to IgE can be detected in sera of individuals with atopic disease, but occasionally elevated levels are also found in sera of normal individuals. During the last years we studied therefore the functional properties of such autoantibodies. Depending on the studied in vitro system one can always detect minimally two different antibody types. Antibodies have been found that either trigger or inhibit mediator release from basophils, that enhance or inhibit binding of IgE to the low IgE receptor and either stimulate or inhibit human IgE synthesis. Based on such in vitro experiments one may conclude that also in vivo autoantibodies exist that either neutralize IgE or have no effect on IgE mediated clinical events. Thus, anti-IgE autoantibodies may hide IgE as for example in those bee sting allergic individuals where we could not detect specific IgE but found IgE hidden within immune complexes, suggesting that the biological activity of IgE was not neutralized. A similar phenomenon may exist in asthmatic individuals. In a recent study we found that non-atopic asthmatic children had indeed low levels of serum IgE, but showed the same levels of autoantibodies to IgE, against suggesting that IgE was hidden within immune complexes. Thus, our ongoing research addresses the question whether in diseases of unclear atopic origin IgE may nevertheless play a critical role but based on possible artefacts in the IgE detection assays some of the clinically relevant IgE may escape.


Pregnancy outcomes in patients with systemic autoimmunity.
The impact of maternal systemic autoimmune diseases on pregnancy outcome is not unequivocally defined. We analysed the pregnancy outcome of 221 pregnancies from 181 autoimmune patients, consecutively followed in a single Italian reference centre from 2001 to 2009. All patients were prospectively followed with monthly visits. Pregnancy outcome was compared with the previous obstetrical history. The patient population comprised five groups: primary antiphospholipid syndrome (PAPS, 39 pregnancies), antiphospholipid syndrome associated with a rheumatic disease (APS/RD, 17 pregnancies), other RD (92 pregnancies), isolated autoantibodies (autoAbs) in the absence of a definite autoimmune disease (aAbs, 38 pregnancies) and reactive arthritis or spondyloarthropathies (35 pregnancies). Of these patients, 50.6% had previous pregnancy complications with an anamnestic live-birth rate of 43.4%. In these patients, complications dropped to 28.2% (44/156). This percentage was very similar to that observed in the 221 pregnancies (29.9%, 66/221) with a live-birth rate of 87.3%. Mean neonatal weight was 3018 ± 611 g; mean gestational age at delivery was 38.17 ± 2.79 weeks. Thus, 10.4% of pregnancies resulted in preterm delivery and 10.9% newborns had low weight at delivery. APS/RD patients had the worse outcome: 17.6% resulted in miscarriage, 14.3% resulted in growth restriction and 50% resulted in preterm delivery. This result was mainly due to patients with APS/systemic lupus erythematosus (SLE) that had the lowest gestational age at delivery (30.8 ± 3.56 weeks) and the lowest newborn weight (1499 ± 931 g). Results confirm that a strict follow-up and targeted treatments significantly improve pregnancy outcomes in autoimmune patients with PAPS, SLE and isolated autoAbs. The pregnancy outcome in patients with APS/SLE remains unsatisfactory.


[Intracranial MR flow signal changes in extracranial occlusions of the internal carotid artery].
Spin-echo sequences allow an identification of the basal brain vessels because of their typical signal void. The increasing use of MRI also in clarification of cerebrovascular disease requires a good knowledge of intracranial flow alterations. We saw characteristic signal pattern in the carotid siphon in 22 prospective examined patients with occlusion of the internal carotid artery confirmed by ultrasound. The lower siphon showed a loss of flow void and a brain isointense signal. The signal in the upper siphon was identical or showed a flow void with significant calibre reduction, probably because of collateral filling. This diagnostic potential of MRI is a remarkable advantage in comparison to CT and should be routinely considered in patients with cerebrovascular disease.


The slack test does not assess maximal shortening velocity of muscle fascicles in humans.
The application of a series of extremely high accelerative motor-driven quick releases while muscles contract isometrically (i.e. slack test) has been proposed to assess unloaded velocity in human muscle. This study aimed to measure gastrocnemius medialis fascicle shortening velocity (VF) and tendinous tissue shortening velocity during motor-driven quick releases performed at various activation levels to assess the applicability of the slack test in humans. Gastrocnemius medialis peak VF and joint velocity recorded from 25 participants using high frame rate ultrasound during quick releases (at activation levels from 0% to 60% of maximal voluntary isometric torque) and during fast contractions without external load (ballistic condition) were compared. Unloaded joint velocity calculated using the slack test method increased whereas VF decreased with muscle activation level (P≤0.03). Passive and low-level quick releases elicited higher VF values (≥41.8±10.7 cm s-1) compared with the ballistic condition (36.3±8.7 cm s-1), while quick releases applied at 60% of maximal voluntary isometric torque produced the lowest VF These findings suggest that initial fascicle length, complex fascicle-tendon interactions, unloading reflex and motor-driven movement pattern strongly influence and limit the shortening velocity achieved during the slack test. Furthermore, VF elicited by quick releases is likely to reflect substantial contributions of passive processes. Therefore, the slack test is not appropriate to assess maximal muscle shortening velocity in vivo.


[Malignant intracranial germ cell tumor treated with chemotherapy and radiotherapy without histopathological confirmation].
In two men aged 19 and 24 years, a rare malignant intracranial germ cell tumour was diagnosed in the pineal gland region and in the second patient in a suprasellar position as well. Although histological confirmation is required in general to diagnose a malignancy, obtaining it can be difficult and potentially dangerous in case of intracranial neoplasms. The clinical picture of a young patient with an intracerebral tumour localised in the midline of the brain and increased levels of the tumour markers alpha-foetoprotein and/or human chorion gonadotrophin (beta-HCG) in blood and/or CSF makes any other diagnosis highly unlikely. There is no place for radical surgery in the first-line treatment of malignant intracerebral germ cell tumours because of the sensitivity to radio- and chemotherapy. Also, the sensitivity to chemotherapy makes it possible to reduce radiation volume and dose in an effort to avoid the serious complications of craniospinal irradiation. Both patients responded well to chemotherapy based on cisplatin followed by radiotherapy.


Social support and thriving health: a new approach to understanding the health of indigenous Canadians.
We examined the importance of social support in promoting thriving health among indigenous Canadians, a disadvantaged population. We categorized the self-reported health status of 31625 adult indigenous Canadians as thriving (excellent, very good) or nonthriving (good, fair, poor). We measured social support with indices of positive interaction, emotional support, tangible support, and affection and intimacy. We used multivariable logistic regression analyses to estimate odds of reporting thriving health, using social support as the key independent variable, and we controlled for educational attainment and labor force status. Compared with women reporting low levels of social support, those reporting high levels of positive interaction (odds ratio [OR]=1.4; 95% confidence interval [CI]=1.2, 1.6), emotional support (OR=2.1; 95% CI=1.8, 2.4), and tangible support (OR = 1.4; 95% CI = 1.2, 1.5) were significantly more likely to report thriving health. Among men, only emotional support was significantly related to thriving health (OR=1.7; 95% CI=1.5, 1.9). Thriving health status was also significantly mediated by age, aboriginal status (First Nations, Métis, or Inuit), educational attainment, and labor force status. Social support is a strong determinant of thriving health, particularly among women. Research that emphasizes thriving represents a positive and necessary turn in the indigenous health discourse.


Hemodynamic assessment of the circulation in 200 normal hands.
The frequent use of arterial puncture, cannulation, and A-V shunting has demonstrated a greater degree of safety than could be predicted from previous anatomic and angiographic studies. Using a noninvasive technique, the status of the superficial palmar arch (SPA) and relative contributions of radial and ulnar arteries were determined in 100 volunteers with no history of vascular disease. Although there was no significant difference in the diameter of the vessels at the wrist, the flows showed statistically significant ulnar dominance, suggesting that the difference is a factor of a lower distal resistance on the ulnar side. The lower resistance across the ulnar bed probably is responsible for the clinically observed ease of sacrifice of the radial artery contribution to palmar flow. The SPA was found to be incomplete in 11.0% of the hands.


Activation of the epithelial-to-mesenchymal transition factor snail mediates acetaldehyde-induced intestinal epithelial barrier disruption.
Acetaldehyde (AcH) is mutagenic and can reach high concentrations in colonic lumen after ethanol consumption and is associated with intestinal barrier dysfunction and an increased risk of progressive cancers, including colorectal carcinoma. Snail, the transcription factor of epithelial-mesenchymal transition, is known to down-regulate expression of tight junction (TJ) and adherens junction (AJ) proteins, resulting in loss of epithelial integrity, cancer progression, and metastases. As AcH is mutagenic, the role of Snail in the AcH-induced disruption of intestinal epithelial TJs deserves further investigation. Our aim was to investigate the role of oxidative stress and Snail activation in AcH-induced barrier disruption in Caco-2 monolayers. The monolayers were exposed from the apical side to AcH ± L-cysteine. Reactive oxygen species (ROS) generation and Snail activation were assessed by ELISA and immunofluorescence. Paracellular permeability, localization, and expression of ZO-1, occludin, E-cadherin, and β-catenin were examined using transepithelial electrical resistance (TEER), fluorescein isothiocyanate-labeled dextran 4 kDa (FITC-D4), immunofluorescence, and ELISA, respectively. Involvement of Snail was further addressed by inhibiting Snail using small interfering RNA (siRNA). Exposure to 25 μM AcH increased ROS generation and ROS-dependently induced Snail phosphorylation. In addition, AcH increased paracellular permeability (decrease in TEER and increase in FITC-D4 permeation) in association with redistribution and decrease of TJ and AJ protein levels, which could be attenuated by L-cysteine. Knockdown of Snail by siRNA attenuated the AcH-induced redistribution and decrease in the TJ and AJ proteins, in association with improvement of the barrier function. Our data demonstrate that oxidative stress-mediated Snail phosphorylation is likely a novel mechanism contributing to the deleterious effects of AcH on the TJ and AJ, and intestinal barrier function.


Glucagon and vasopressin interactions on Ca2+ movements in isolated hepatocytes.
The effects of glucagon and vasopressin, singly or together, on cytosolic free Ca2+ concentration [( Ca2+]i) and on the 45Ca2+ efflux were studied in isolated rat liver cells. In the presence of 1 mM external Ca2+, glucagon and vasopressin added singly induced sustained increases in [Ca2+]i. The rate of the initial fast phase of the [Ca2+]i increase and the magnitude of the final plateau were dependent on the concentrations (50 pm-0.1 microM) of glucagon and vasopressin. Preincubating the cells with a low concentration of glucagon (0.1 nM) for 2 min markedly accelerated the fast phase and elevated the plateau of the [Ca2+]i increase caused by vasopressin. In the absence of external free Ca2+, glucagon and vasopressin transiently increased [Ca2+]i and stimulated the 45Ca2+ efflux from the cells, indicating mobilization of Ca2+ from internal store(s). Preincubating the cells with 0.1 nM-glucagon accelerated the rate of the fast phase of the [Ca2+]i rise caused by the subsequent addition of vasopressin. However, unlike what was observed in the presence of 1 mM-Ca2+, glucagon no longer enhanced the maximal [Ca2+]i response to vasopressin. In the absence of external free Ca2+, higher concentrations (1 nM-0.1 microM) of glucagon, which initiated larger increases in [Ca2+]i, drastically decreased the subsequent Ca2+ response to vasopressin (10 nM). At these concentrations, glucagon also decreased the vasopressin-stimulated 45Ca2+ efflux from the cells. It is suggested that, in the liver, glucagon accelerates the fast phase and elevates the plateau of the vasopressin-mediated [Ca2+]i increase respectively by releasing Ca2+ from the same internal store as that permeabilized by vasopressin, probably the endoplasmic reticulum, and potentiating the influx of extracellular Ca2+ caused by this hormone.


Cold-pressed minke whale oil reduces circulating LDL/VLDL-cholesterol, lipid oxidation and atherogenesis in apolipoprotein E-deficient mice fed a Western-type diet for 13 weeks.
Long-chain n3-polyunsaturated fatty acids (LC n3-PUFA) are well known for their anti-inflammatory activity and their impact on cardiovascular disease. Cold-pressed whale oil (CWO) has half the amount of LC n3-PUFA compared to cod liver oil (CLO). Still, there has been observed more pronounced beneficial effects on cardiovascular disease markers from intake of CWO compared to intake of CLO in human intervention studies. Extracts from CWO deprived of fatty acids have also been shown to display antioxidative and anti-inflammatory effects in vitro. The aim of this study was to investigate whether intake of a high-fat Western-type diet (WD) supplemented with CWO would prevent the development of atherosclerotic lesions in apolipoprotein E-deficient (ApoE-/-) mice. Seventy female ApoE-/- mice were fed a WD containing 1% CWO, CLO or corn oil (CO). Atherosclerotic lesion formation, body and tissue weights, hepatic gene expression together with serum levels of LDL/VLDL-cholesterol, ox-LDL, total antioxidant status and various serum cardiovascular disease/proinflammatory markers were evaluated. Statistical analyses were performed using SPSS, and Shapiro-Wilk's test was performed to determine the distribution of the variables. Statistical difference was assessed using One-Way ANOVA with Tukeys' post hoc test or Kruskal-Wallis test. The hepatic relative gene expression was analysed with REST 2009 (V2.0.13). Mice fed CWO had less atherosclerotic lesions in the aortic arch compared to mice fed CO. Levels of LDL/VLDL-cholesterol and ox-LDL-cholesterol were also markedly reduced whereas total antioxidant levels were enhanced in mice fed CWO compared to CO-fed mice. In addition, CWO-fed mice gained less weight and several hepatic genes involved in the cholesterol metabolism were up-regulated compared to CO-fed mice. In the present study mice fed a WD supplemented with 1% CWO had reduced formation of atherosclerotic lesions in the aortic arch, reduced serum LDL/VLDL-cholesterol and ox-LDL-cholesterol, increased serum total antioxidant status and reduced body weight compared to mice fed a WD supplemented with 1% CO.


Sequencing and Genomic Diversity Analysis of IncHI5 Plasmids.
IncHI plasmids could be divided into five different subgroups IncHI1-5. In this study, the complete nucleotide sequences of seven bla IMP- or bla VIM-carrying IncHI5 plasmids from Klebsiella pneumoniae, K. quasipneumoniae, and K. variicola were determined and compared in detail with all the other four available sequenced IncHI5 plasmids. These plasmids carried conserved IncHI5 backbones composed of repHI5B and a repFIB-like gene (replication), parABC (partition), and tra1 (conjugal transfer). Integration of a number of accessory modules, through horizontal gene transfer, at various sites of IncHI5 backbones resulted in various deletions of surrounding backbone regions and thus considerable diversification of IncHI5 backbones. Among the accessory modules were three kinds of resistance accessory modules, namely Tn10 and two antibiotic resistance islands designated ARI-A and ARI-B. These two islands, inserted at two different fixed sites (one island was at one site and the other was at a different site) of IncHI5 backbones, were derived from the prototype Tn3-family transposons Tn1696 and Tn6535, respectively, and could be further discriminated as various intact transposons and transposon-like structures. The ARI-A or ARI-B islands from different IncHI5 plasmids carried distinct profiles of antimicrobial resistance markers and associated mobile elements, and complex events of transposition and homologous recombination accounted for assembly of these islands. The carbapenemase genes bla IMP-4, bla IMP-38 and bla VIM-1 were identified within various class 1 integrons from ARI-A or ARI-B of the seven plasmids sequenced in this study. Data presented here would provide a deeper insight into diversification and evolution history of IncHI5 plasmids.


[Interpersonal competence in caring of people with diabetes: perception of nurses].
This is a qualitative and exploratory study which aimed to apprehend the perceptions of nurses who care for people with Diabetes on the interpersonal competence. The subjects were eleven nurses who performed their activities in the Units of Family Health in the urban area, and completed a consent form. The data, obtained through semi-structured interview, were analyzed and categorized by thematic analysis. The results showed that nurses perceive the interpersonal competence as an ability to interact with the patient, as the establishment of an effective interpersonal relationship and as forms of interpersonal relationships. It appeared that the exercise of that power is not subject to standardization, what emphasizes the uniqueness of the processes of interaction and of health care.


Deconstructing Chronic Low Back Pain in the Older Adult-Step by Step Evidence and Expert-Based Recommendations for Evaluation and Treatment: Part X: Sacroiliac Joint Syndrome.
To present an algorithm of sequential treatment options for managing sacroiliac joint (SIJ) pain in the setting of chronic low back pain (CLBP) in the older adult. This is the tenth part in a series, and includes an illustrative clinical case. The stepped care drug table and evaluation and treatment algorithm were created following a thorough literature review of approaches and subsequent analysis through a modified Delphi process. The principal investigator developed the initial draft, which was refined for content by an interdisciplinary panel of five experts. The refined materials were then assessed for the feasibility of implementation and validity of recommendations for older adults in a primary care setting by a panel of nine primary care providers. While not exclusive to Veteran's Health Administration (VHA) facilities, an emphasis was made to include resources and medications available to providers in the VHA. The algorithm and drug table developed to systematically identify and address SIJ pain in the older adult is presented here. The process should begin with recognizing the presenting symptoms of CLBP stemming from the SI region, and supporting physical exam testing using the compression test and thigh thrust maneuver. Identification of the SIJ as a pain generator is followed by assessment and treatment of contributory factors. SIJ pain treatment should begin with education and self-management including exercise, and may escalate to include interventional procedures and/or referral to a pain rehabilitation program. Pain originating from the SIJ is often under-recognized, but a structured and consistent approach can help identify older patients who would benefit from treatment of this contributor to CLBP. Chronic Low Back Pain; Sacroiliac Joint Pain; Older Adults.


Benzonatate Safety and Effectiveness: A Systematic Review of the Literature.
To review the available literature regarding the treatment effects and efficacy of benzonatate needed to better inform patients, providers, and regulators evaluating its role in modern medical therapies. Comprehensive literature searches were conducted in PubMed, Embase (Elsevier), Cochrane Library, and Scopus for original research articles evaluating the effectiveness, tolerability, and safety profile of benzonatate from January 1956 through August 2022. The identified studies were screened for relevance and then assessed for inclusion through a full-text review, data extraction, and quality assessment by multiple reviewers using the online software Covidence. The selection process resulted in 37 articles consisting of 21 cohort studies, 5 experimental studies, and 11 case studies and series. Initial clinical studies exploring potential therapeutic benefits collected data from very small populations and limited clinical settings. Safety is primarily assessed in terms of toxicity due to overdose or inappropriate use. Quality assessment raised concerns for high degrees of biases primarily related to the limited sample size, data collection, generalizability, and study design. This review reveals substantial limitations within existing evidence pertaining to the safety and clinical effectiveness of benzonatate and thus, a need for large observational studies or randomized trials to better characterize its role and value in modern medical practice. Rising safety concerns should bring closer scrutiny upon the prescription of benzonatate whose approval is founded upon evidence that would not stand up to current regulatory review.


Chromatin regulator SMARCAL1 modulates cellular lipid metabolism.
Biallelic mutations of the chromatin regulator SMARCAL1 cause Schimke Immunoosseous Dysplasia (SIOD), characterized by severe growth defects and premature mortality. Atherosclerosis and hyperlipidemia are common among SIOD patients, yet their onset and progression are poorly understood. Using an integrative approach involving proteomics, mouse models, and population genetics, we investigated SMARCAL1's role. We found that SmarcAL1 interacts with angiopoietin-like 3 (Angptl3), a key regulator of lipoprotein metabolism. In vitro and in vivo analyses demonstrate SmarcAL1's vital role in maintaining cellular lipid homeostasis. The observed translocation of SmarcAL1 to cytoplasmic peroxisomes suggests a potential regulatory role in lipid metabolism through gene expression. SmarcAL1 gene inactivation reduces the expression of key genes in cellular lipid catabolism. Population genetics investigations highlight significant associations between SMARCAL1 genetic variations and body mass index, along with lipid-related traits. This study underscores SMARCAL1's pivotal role in cellular lipid metabolism, likely contributing to the observed lipid phenotypes in SIOD patients.


The hunt for new tuberculosis vaccines: anti-TB immunity and rational design of vaccines.
Tuberculosis (TB) remains to be a leading infectious cause of death worldwide. Apparently, the current BCG vaccine that has been used for 80 years, has failed to control the TB epidemic. Hunting for improved TB vaccine formulations represents a daunting task to TB research community. Anti-TB host defense requires T cell-mediated immunity and we are in desperate need of enhanced understanding of how to develop a new generation of TB vaccines that are able to provoke potent and long-lasting protective cell-mediated immunity, different from almost all of the vaccines currently in use. It is of importance to successful TB vaccine development to identify the key cellular and molecular events governing the generation of anti-TB immunity, but unfortunately little has been understood as to why 90% of infected humans never develop active TB. However, waiting would not help us to win the battle and an ever-intensifying effort is being made to develop various new formulations according to the immunology that we have been learning, in large part, from experimental models. This review article attempts to unite the current understanding of anti-TB immunity with the rational design of anti-TB vaccines. It examines what may have confounded the immunogenicity of current BCG vaccine and the major obstacles to successful development of TB vaccines. It also discusses about antigen presentation, activation of Th1 and Tc1 cells, anti-TB immune effectors and the generation of memory T cells. The vaccine section describes four types of major TB vaccines under development: mycobacterial-, subunit-, plasmid DNA- and viral-based vaccines. A special section is dedicated to the rationale and current design of cytokine-based adjuvant formulations for TB vaccines. We also take this opportunity to introduce our recent development in cytokine transgene adjuvanted BCG vaccination and recombinant adenoviral-based TB vaccines.


The association of personality polygenic risk score, psychosocial protective factors and suicide attempt in mood disorder.
Some personality traits, especially neuroticism, has been found to be associated with suicide attempt (SA) in mood disorder patients. The present study explored the association between personality traits and SA using polygenic risk scores (PRS) for personality among patients with mood disorders. We also investigated the effects of a variety of psychosocial variables on SA. Patients with bipolar disorder (BPD, N = 841) and major depressive disorder (MDD, N = 710) were recruited from hospitals in Taiwan. Lifetime SA and information on psychosocial factors was collected. We calculated the PRS of neuroticism and extraversion. A trend test for SA was performed across quartiles of the PRS for neuroticism and extraversion, and logistic regression analyses were performed to examine the associations between psychosocial factors and SA, accounting for the PRS of personality traits. The prevalence of SA was higher in MDD than in BPD patients. The risk of SA was elevated in MDD patients with a higher quintile of PRS in neuroticism and a lower quintile of PRS in extraversion. The multiple regression analysis results demonstrated that later age of onset, higher family support and resilience, and lower overall social support were protective factors against SA. From the perspective of suicide prevention efforts, strengthening family support and conducting resilience training for patients with mood disorders may be beneficial interventions in clinical settings.


Finger cold-induced vasodilation: a review.
Cold-induced vasodilation (CIVD) in the finger tips generally occurs 5-10 min after the start of local cold exposure of the extremities. This phenomenon is believed to reduce the risk of local cold injuries. However, CIVD is almost absent during hypothermia, when survival of the organism takes precedence over the survival of peripheral tissue. Subjects that are often exposed to local cold (e.g. fish filleters) develop an enhanced CIVD response. Also, differences between ethnic groups are obvious, with black people having the weakest CIVD response. Many other factors affect CIVD, such as diet, alcohol consumption, altitude, age and stress. CIVD is probably caused by a sudden decrease in the release of neurotransmitters from the sympathetic nerves to the muscular coat of the arterio-venous anastomoses (AVAs) due to local cold. AVAs are specific thermoregulatory organs that regulate blood flow in the cold and heat. Their relatively large diameter enables large amounts of blood to pass and convey heat to the surrounding tissue. Unfortunately, information on the quantity of AVAs is lacking, which makes it difficult to estimate the full impact on peripheral blood flow. This review illustrates the thermospecificity of the AVAs and the close link to CIVD. CIVD is influenced by many parameters, but controlled experiments yield information on how CIVD protects the extremities against cold injuries.


Population-based study of the contribution of weather to hip fracture seasonality.
Previous studies have demonstrated seasonal increases in hip fracture incidence, but none have directly assessed the influence of inclement weather on this seasonality. In this study, the daily occurrence of hip fracture among women aged 45 years and older in Rochester, Minnesota, from 1952 through 1989 was ascertained using the resources of the Rochester Epidemiology Project and compared with the occurrence of inclement weather as recorded in hourly readings by the National Weather Service in Rochester for the same time period. Among the women aged 45-74 years, the risk of hip fracture was increased on days with snow (relative risk (RR) = 1.41, 95% confidence interval (CI) 1.10-1.81) or freezing rain (RR = 1.82, 95% CI 1.27-2.62). In this group, the elevated risk of hip fracture in winter, compared with summer (RR = 1.44, 95% CI 1.0-2.09), was reduced after controlling for weather (RR = 1.16, 95% CI 0.81-1.65). Among women aged 75 years and older, ice and snow were not strongly related to fracture occurrence. The winter-related increase in risk (RR = 1.16, 95% CI 0.96-1.40) was essentially unchanged after controlling for weather and was similar to the weather-adjusted seasonality of hip fracture occurrence in younger women. These data suggest that there are factors other than weather that may be linked to the seasonal pattern in hip fracture occurrence and that operate at all ages.


Giardia gene predicts a 183 kDa nucleotide-binding head-stalk protein.
Previously described extended proteins from the cytoskeleton of Giardia lamblia (beta-giardin, median body protein) have been found to be segmented coiled coils with regular structural repeat patterns in their amino acid sequences. Screening a lambda ZAPII library derived from Giardia genomic DNA with an antibody directed against a 34 x 10(3) M(r) giardin isoform selected a gene encoding a much larger polypeptide chain (HPSR2), the sequence of which was determined by chromosome walking the open reading frame. The complete gene has been cloned and expressed as a recombinant protein of 183 x 10(3) M(r). The predicted amino acid sequence of the protein has identifiable features suggesting that it might be a motor protein with an amino-terminal hydrolytic domain attached to a long coiled coil stalk. The presumed head domain is 211 residues and contains a P-loop sequence conserved in purine nucleotide-binding proteins. The remaining 1409 amino acids mainly make up a region of heptad repeats such as in myosin or the kinesin stalk, ending in a small (67 amino acids) carboxy-terminal domain. Fourier analysis of the predicted stalk shows the presence of a strong physical repeat created by regular heptad phase changes dividing the coil into segments of 25 residues. This structure most closely resembles the smaller microtubule-associated median body protein which has segments of 24 residues.


[Validation of an eating attitude scale in a French-speaking Quebec population].
The Eating Attitudes Test (EAT-26) is one of the most frequently used screening questionnaires for anorexia and bulimia nervosa for use with clinical and general populations. Although the psychometric qualities of the instrument have been reported for the English version, little has been done to date to validate a French version. A french version of the EAT-26 was distributed to anglophone students and francophone students, as well as anglophone and francophone patients. Overall, the results demonstrated that the authors' French version has the same psychometric characteristics as the English version when used with clinical and non clinical populations. The overall test and its subscales differentiated between patients and non patients similarly in both linguistic groups. The internal consistency of the two versions, as assessed by Cronbach's alpha, was comparable. Among female high-school students, 14.1% of the anglophone students and 14.3% of the francophone students scored above the cutoff score for significant eating disorders behaviour.