GABA(A) alpha-1 subunit mediated desynchronization of elevated low frequency oscillations alleviates specific dysfunction in stroke--a case report.
The paradoxical effects of the hypnotic imidazopyridine zolpidem, widely reported in persistent vegetative state, have been replicated recently in brain-injured and cognitively impaired patients. However, the neuronal mechanisms underlying these benefits are yet to be demonstrated. We implemented contemporary neuroimaging methods to investigate sensorimotor and cognitive improvements, observed in stroke patient JP following zolpidem administration. We used Magnetic-Resonance-Imaging (MRI) and Magnetic-Resonance-Spectroscopy (MRS) to anatomically and chemically characterize stroke damage. Single-photon-emission-computed-tomography (SPECT) and magnetoencephalography (MEG) were used to identify changes in cerebrovascular perfusion and neuronal network activity in response to sub-sedative doses of zolpidem, zopiclone and placebo. Cognitive improvements were measured using the WAIS-III and auditory-verbal tasks. MRI and MRS revealed a lesion with complete loss of neuronal viability in the left temporal-parietal region; whilst SPECT indicated improved perfusion in the affected hemisphere following zolpidem. MEG demonstrated high-amplitude theta (4-10 Hz) and beta (15-30 Hz) oscillations within the peri-infarct region, which reduced in power coincident with zolpidem uptake and improvements in cognitive and motor function. In JP, functional deficits and pathological oscillations appear coincidentally reduced following administration of low-dose zolpidem. GABA(A) alpha-1 sensitive desynchronisation of pathological oscillations may represent a biomarker and potential therapeutic target in brain injury.


Admission Screening of Methicillin-Resistant Staphylococcus aureus with Rapid Molecular Detection in Intensive Care Unit: A Three-Year Single-Centre Experience in Hong Kong.
Background. The admission screening of methicillin-resistant Staphylococcus aureus (MRSA) by rapid molecular assay is considered to be an effective method in reducing the transmission of MRSA in intensive care unit (ICU). Method. The admission screening on patients from ICU once on their admissions by BD GeneOhm MRSA assay has been introduced to Prince of Wales Hospital, Hong Kong, since 2008. The assay was performed on weekdays and reported on the day of testing. Patients pending for results were under standard precautions until the negative screening results were notified, while contact precautions were implemented for MRSA-positive patients. In this study, we compared the MRSA transmission rate in molecular screening periods (2008 to 2010) with the historical culture periods (2006 to 2007) as control. Results. A total of 4679 samples were tested; the average carriage rate of MRSA on admission was 4.45%. By comparing with the historical culture periods, the mean incidence ICU-acquired MRSA infection was reduced from 3.67 to 1.73 per 1000 patient bed days. Conclusion. The implementation of admission screening of MRSA with molecular method in intensive care unit could reduce the MRSA transmission, especially in the area with high MRSA prevalence situation in Hong Kong.


Pulmonary injury in rats following continuous exposure to 60% O2 for 7 days.
Morphological, biochemical, and physiological studies were done on rats exposed to 60% O2 for 7 days. This exposure did not induce O2 tolerance but instead caused a significant decrease in survival time of animals subsequently exposed to pure O2. The activity of lung superoxide dismutases and glucose-6-phosphate dehydrogenase were unchanged after exposure to 60% O2. A decrease in lung compliance was suggested by changes in the total lung capacity and in the pressure-volume curves of excised lungs. Ventilation of these animals with large tidal excursion resulted in pulmonary edema. Morphometric analyses revealed a significant decrease in alveolar air volume and an increase in the number of alveolar macrophages. The most significant lesions involved the pulmonary vascular bed. The volume and thickness of the capillary endothelium was decreased. There were focal areas of pericapillary fluid accumulations, and a number of the smaller vessels had perivascular edema. These findings suggest that significant pulmonary injury occurs in rats exposed to 60% O2 and that the primary site of injury is the pulmonary capillary endothelium.


Symptom burden in chronic obstructive pulmonary disease and cancer.
Chronic obstructive pulmonary disease (COPD) is a crippling disease with a high worldwide prevalence. The purpose of this review is to highlight recent studies which define the global impact of COPD on quality of life. There are direct implications for care. Dyspnea is a dominant and defining symptom for patients with COPD, but the overall degree of impairment suffered by patients with COPD extends far beyond shortness of breath. A series of recent studies gives us insight into both the physical and the psychosocial burdens of the disease and their negative net effects upon quality of life. The suffering of patients with COPD is similar to that of patients with cancer, and palliative measures have been shown to be an important component of comprehensive care. The symptom burden in patients with severe COPD is high and is comparable to that of patients with cancer. Ironically, patients with COPD could be said to suffer more than those with cancer; the symptom burden is similar, but patients with COPD tend to live longer. The literature is replete with evidence that a palliative care approach to patients with cancer increases the quality of life (and perhaps even the quality of death). The same palliative care approach can and should be used for patients with COPD. There are now objective data to support the benefits of such an approach.


The startle response: developmental effects and a paradigm for children and adults.
A film paradigm was developed to examine baseline and emotion modulated startle across a broad age range from preschool to adulthood. The paradigm was tested in children (3-, 5-, 7-, and 9-year-olds) and adults (total N = 122). The paradigm elicited a similar startle potentiation pattern across age groups; however, baseline startle changed with age: 3- and 5-year-olds showed lower response probability and magnitude of baseline startle than adults. Females exhibited larger baseline startle response probability and overall magnitude than did males; however, no sex by emotion modulated startle interaction was noted. Anxiety measures were obtained for all children. Individual differences in anxiety were associated with baseline startle magnitude among older but not younger children. No association of anxiety with startle potentiation was noted. Overall the film paradigm was applicable across a wide developmental span, revealing potential developmental and gender differences in baseline startle magnitude and response probability.


Superoxide scavenging and Akt inhibition in myocardium ameliorate pressure overload-induced NF-κB activation and cardiac hypertrophy.
Recent studies from our laboratory and others have shown that increases in cytoplasmic superoxide (O(2)(·-)) levels and Akt activation play a key role in agonist-stimulated NF-κB activation and cardiomyocyte hypertrophy in vitro. In this study, we tested the hypothesis that adenovirus (Ad)-mediated intramyocardial gene transfer of cytoplasmic superoxide dismutase (AdCu/ZnSOD) or a dominant-negative form of Akt (AdDNAkt) in mice would attenuate pressure overload-induced increases in activation of the redox-sensitive transcription factor NF-κB and cardiac hypertrophy. Adult C57BL/6 mice were subjected to thoracic aortic banding (TAB) or sham surgery, and intramyocardial injections of viral vectors (AdCu/ZnSOD, AdDNAkt, or control) were performed. There was robust transgene expression in the heart, which peaked 6-7 days after injection and then declined to undetectable levels by 12-14 days. In mice injected with AdBgL II, TAB caused a significant increase in O(2)(·-) generation and cardiac mass at 1 wk, and these responses were markedly attenuated by AdCu/ZnSOD. In addition, TAB induced time-dependent activation of NF-κB in the myocardium as measured longitudinally by in vivo bioluminescent imaging of NF-κB-dependent luciferase expression. This was also abolished by intracardiac AdCu/ZnSOD or AdDNAkt, but not the control vector. The inhibition of Akt and O(2)(·-)-mediated NF-κB activation in TAB hearts was associated with an attenuation of cardiac hypertrophy. Since a direct cause-and-effect relationship between NF-κB activation and cardiomyocyte hypertrophy has been established previously, our data support the hypothesis that increased O(2)(·-) generation and Akt activation are key signaling intermediates in pressure overload-induced activation of NF-κB and cardiac hypertrophy.


A hybrid, low-cost tissue-like epidural needle insertion simulator.
Epidural and spinal anesthesia are mostly performed "blind" without any medical imaging. Currently, training of these procedures is performed on human specimens, virtual reality systems, manikins and mostly in clinical practice supervised by a professional. In this study a novel hybrid, low-cost patient simulator for the training of needle insertion into the epidural space was designed. The patient phantom provides a realistic force feedback comparable with biological tissue and enables sensing of the needle tip position during insertion. A display delivers the trainee a real-time feedback of the needle tip position.


Balanced gradient boosting from imbalanced data for clinical outcome prediction.
In clinical outcome prediction, such as disease diagnosis and prognosis, it is often assumed that the class, e.g., disease and control, is equally distributed. However, in practice we often encounter biological or clinical data whose class distribution is highly skewed. Since standard supervised learning algorithms intend to maximize the overall prediction accuracy, a prediction model tends to show a strong bias toward the majority class when it is trained on such imbalanced data. Therefore, the class distribution should be incorporated appropriately to learn from imbalanced data. To address this practically important problem, we proposed balanced gradient boosting (BalaBoost) which reformulates gradient boosting to avoid the overfitting to the majority class and is sensitive to the minority class by making use of the equal class distribution instead of the empirical class distribution. We applied BalaBoost to cancer tissue diagnosis based on miRNA expression data, premature death prediction for diabetes patients based on biochemical and clinical variables and tumor grade prediction of renal cell carcinoma based on tumor marker expressions whose class distribution is highly skewed. Experimental results showed that BalaBoost outperformed the representative supervised learning algorithms, i.e., gradient boosting, Random Forests and Support Vector Machine. Our results led us to the conclusion that BalaBoost is promising for clinical outcome prediction from imbalanced data.


Memory CD44(int) CD8 T cells show increased proliferative responses and IFN-gamma production following antigenic challenge in vitro.
F5 TCR transgenic mice challenged in vivo with peptide generate long-lived primed CD8 T cells that hyper-proliferate in response to peptide in vitro. These primed CD8 T cells can be subdivided into three distinct populations on the basis of CD44 cell surface expression. In this report, we show that among primed CD8 T cells, those expressing intermediate levels of CD44 appear to be true memory T cells by the measurement of a variety of characteristics. Indeed, these cells hyper-proliferate in response to peptide re-stimulation in vitro, and produce IFN-gamma with faster kinetics and at higher levels than naive populations in vitro. We also show that CD8 T cells expressing high levels of CD44 express several activation markers and cycle in vivo in the absence of antigen. However, this population is unable to respond to peptide stimulation in vitro as measured by both proliferation and IFN-gamma secretion. The origin and specificity of these cells is unknown. These results provide evidence that memory CD8 T cells are functionally different from naive CD8 T cells both in terms of proliferation and cytokine secretion. They identify the CD8/CD44(int) T cells as the population responsible for hyper-reactivity in vitro.


Adsorption-desorption behavior of acetochlor to soils in the presence of some environmental substances.
The behavior of herbicide acetochlor adsorption-desorption to soil in the presence of humic acid (HA), anionic surfactant sodium dodecylbenzene sulfonate (SDBS), cationic surfactant hexadecyltrimethyl-ammonium bromide (HDAB) and NH4NO3 as a chemical fertilizer was studied. Observed acetochlor adsorption isotherm were well described using Freundlich isotherm equation, from which the desorption isotherm equation has been deduced. The deduced equation can more directly describe acetochlor desorption process. The results showed that the enhance of acetochlor adsorption capacity by solid HA was greater than by soluble HA. The presence of NH4 NO3 can slightly enhance acetochlor adsorption to soil by comparison with that measured in NH4 NO3-free solution. In soil-water system, surfactant-acetochlor interaction is very complex, and the surfactant adsorptions as well as acetochlor adsorption need to be considered. When acetochlor-soil suspensions contained lower concentration SDBS or HDAB (40 mg/L), Kf for acetochlor adsorption was decreased in comparison to that measured in SDBS- or HDAB-free solution. When acetochlor-soil suspensions contained higher concentration SDBS or HDAB (corresponding 1400 mg/L or 200 mg/L), Kf for acetochlor adsorption was increased in comparison to that measured in SDBS- or HDAB-free solution.


An explanatory machine learning framework for studying pandemics: The case of COVID-19 emergency department readmissions.
One of the major challenges that confront medical experts during a pandemic is the time required to identify and validate the risk factors of the novel disease and to develop an effective treatment protocol. Traditionally, this process involves numerous clinical trials that may take up to several years, during which strict preventive measures must be in place to control the outbreak and reduce the deaths. Advanced data analytics techniques, however, can be leveraged to guide and speed up this process. In this study, we combine evolutionary search algorithms, deep learning, and advanced model interpretation methods to develop a holistic exploratory-predictive-explanatory machine learning framework that can assist clinical decision-makers in reacting to the challenges of a pandemic in a timely manner. The proposed framework is showcased in studying emergency department (ED) readmissions of COVID-19 patients using ED visits from a real-world electronic health records database. After an exploratory feature selection phase using genetic algorithm, we develop and train a deep artificial neural network to predict early (i.e., 7-day) readmissions (AUC = 0.883). Lastly, a SHAP model is formulated to estimate additive Shapley values (i.e., importance scores) of the features and to interpret the magnitude and direction of their effects. The findings are mostly in line with those reported by lengthy and expensive clinical trial studies.


Factors Associated With Low Physical Activity in Two Latin American Populations at Risk of Developing Type 2 Diabetes: An Exploratory Analysis.
Low level of physical activity is a risk factor for chronic non-communicable diseases. Specifically, people at risk of Type 2 Diabetes (T2D) have shown to benefit from being physically active. The objective of this study was to explore what factors were associated with low physical activity in people at high risk of T2D living in Bogota and Barranquilla, Colombia. Methodology: Cross-sectional study using baseline data from a quasi-experimental clinical trial (PREDICOL Project). The study included 1,135 participants of Bogota and Barranquilla that presented a high risk of developing T2D according to the Finnish Diabetes Risk Score (>12 points) and who underwent an oral glucose tolerance test. The main outcome variable was the level of physical activity assessed by the International Physical Activity Questionnaire. Unadjusted and adjusted logistic regression analysis were used to calculate odds ratios (OR) and the corresponding 95% confidence intervals (CI). Results: In total, 72.5% of the study participants had low level of physical activity. Participants in the age group between 45 and 54 years showed 74% greater odds of having low physical activity compared with the youngest age group (OR 1.74, 95% CI 1.1 -2.8). People living in Barranquilla were eight times more likely to have low physical activity compared with those in Bogotá D.C. (OR 8.1, 95% CI 5.7 to 11.4). Conclusion: A large proportion of the population at risk of developing D2T in two large cities of Colombia have a sedentary lifestyle. Interventions should be designed and implemented in order to increase physical activity in these populations.


GLP-1 agonist liraglutide improves ouabain-induced mania and depressive state via GSK-3β pathway.
Bipolar disorder (BD) is a severe mental illness characterized by aberrant mood changes between hypomania and mania or mixed states and depression. Metabolic changes also accompany disease progression and cause significant morbidity. Symptomatic treatment options are available, but asymptomatic patients and poor drug responders are significant problems. Based on the most common pharmacological agent that is used in the treatment, lithium and its main mechanisms of action, oxidative stress, and glycogen synthase kinase-3β (GSK-3β) signaling are extensively investigated. However, knowledge about the effects of compounds that positively affect oxidative stress and GSK-3β signaling, such as glucagon-like peptide-1 (GLP-1) mimetics, liraglutide, is still missing. Therefore, in this study, we aimed to investigate the effects of liraglutide on the ouabain-induced bipolar disease model in rats. After intracerebroventricular single dose ouabain administration, animals were treated with 100, 200, and 400 µg/kg liraglutide (s.c.) and valproic acid (200 mg/kg, i.p.) for 10 d. The locomotion and depressive states of animals were assessed by an open field, forced swimming test, and sucrose preference tests. Serum total antioxidant (TAS) and oxidant states (TOS) and glutathione, malonyl dialdehyde (MDA) levels in the brain tissue were determined. GSK-3β phosphorylation was evaluated by western blotting. Our results demonstrated that liraglutide attenuated ouabain-induced hyperlocomotion and depressive state. Additionally, liraglutide prevented oxidative stress after ouabain administration. Decreased GSK-3β phosphorylation due to the ouabain insult was alleviated by liraglutide treatment. These findings indicate that the manic and depressive-like behaviors are ameliorated by liraglutide, which exerted antioxidant action, possibly improving GSK-3β phosphorylation.


Robust superhydrophobic attapulgite meshes for effective separation of water-in-oil emulsions.
Special-wetting materials have been broadly studied in various oil/water separation processes. However, there are granted numerous challenges in extremely durable materials with mechanical robustness, especially considering the great requirements in purifying emulsified oil/water mixtures. Herein, we present a facile route to prepare robust superhydrophobic surface by spraying Octadecyltrimethoxys modified attapulgite suspension combined with inorganic adhesive on mesh. The as-prepared meshes show eminent superhydrophobicity and acquire a gravity driven water-in-oil emulsion separation with efficiency above 99.7% and flux above 106.7 L m-2h-1 even after multiple cycles. In addition, the meshes exhibit robust and stable superhydrophobicity with water contact angles above 150° after 200 sandpaper abrasion (transverse shear force) and harsh sand impact (longitudinal impact force) cycles, and meanwhile, still maintain outstanding emulsion separation performance during tests. This study is meaningful for the development of fabricating low-energy separating materials with high efficiency and mechanical robustness for harsh chemical engineering.


Protein-energy undernutrition and life-threatening complications among the hospitalized elderly.
To determine whether elderly patients with protein-energy undernutrition at admission are at increased risk for subsequent life-threatening events after controlling for illness severity. Prospective cohort study. University-affiliated Department of Veterans Affairs hospital. Five hundred eighty-six nonterminal patients (mean age 74 +/- 6 [SD] years, 98% male, 86% white) with a length of stay of 3 days or more. Life-threatening complications. Subsequent to admission, 37 subjects (6.3%) experienced at least 1 life-threatening complication. All of the putative nutrition variables examined and many non-nutrition, illness severity measures were strongly correlated with the risk of a life-threatening complication by univariate analyses (P <.05 for all analyses). After controlling for illness severity, admission serum albumin, prealbumin, and cholesterol were no longer significantly correlated with the outcome. In contrast, weight loss (>5% within 6 months), body mass index, mid-arm circumference, and suprailiac skinfold thickness remained strong independent predictors. The adjusted relative risk of a life-threatening complication ranged from 2.9 (95% confidence interval [CI], 1.3 to 6.4) for a body mass index <22 kg/m2 to 7.1(95% CI, 2.0 to 25.7) for a suprailiac skinfold thickness in the lower tertile for the study population. The putative nutrition and illness severity variables were highly intercorrelated. There is a complex interrelationship between nutritional status, illness severity, and clinical outcomes among the hospitalized elderly. The serum secretory proteins and cholesterol are correlated with other indicators of illness severity and adverse outcomes, but may not be good markers of nutritional risk. In contrast, weight loss, a low body mass index, and other indicators of lean and fat mass depletion appear to place the patient at increased risk for adverse outcomes independent of illness severity. Whether it is possible to reverse such established nutritional deficits and reduce complication risk in the acute care setting remains to be determined.


Rethinking the Use of Neutral Faces as a Baseline in fMRI Neuroimaging Studies of Axis-I Psychiatric Disorders.
Major Axis-I disorders including major depressive disorder (MDD), bipolar disorder (BD), anxiety disorder, and schizophrenia are associated with a host of aberrations in the way social stimuli are processed. Face perception tasks are often used in neuroimaging research of emotion processing in both healthy and patient populations, and to date, there exists a mounting body of evidence, both behavioral and within the brain, indicating that emotional faces compared to neutral faces are processed abnormally by those with Axis-I disorders relative to healthy control (HC) groups. The use of neutral faces as a "baseline control condition" is predicated on the assumption that neutral faces are processed in the same way HCs and individuals with major Axis-I disorders. In this paper, existing fMRI studies examining the way neutral faces are processed in groups with Axis-I disorders involving socioaffective perception are reviewed. In reviewing available studies, a consistent pattern of results demonstrated that these disorders are associated with abnormal frontolimbic activity in response to neutral faces and in particular within the amygdala and prefrontal regions such as the dorsolateral prefrontal cortex (DLPFC) and anterior cingulate cortex (ACC) compared to HC groups. Specifically, increased amygdala activation was consistently reported in response to neutral faces in anxiety disorders and schizophrenia. Abnormal medial PFC activity was reported in patients with MDD, and patients with BD exhibit decreased activity in the DLPFC and ACC relative to HCs. In addition, specific suggestions to overcome these obstacles with new research and additional analyses are discussed.


Complexities in the genetic dissection of quantitative trait loci.
The analysis of complex traits, including those involved in many common diseases, has encountered significant difficulties, and, despite major efforts during the past decade, has had little success. Current advances in genomics, however, promise to change this. Recently, Steinmetz et al. used a new technique to produce the first complete quantitative trait locus (QTL) analysis of a complex trait from phenotype to gene to be published entirely in a single report. This marks a significant advance over previous QTL analyses, which took several years. The work exemplifies some of the complexities of QTL mapping and demonstrates a novel method to resolve the underlying genetic architecture of QTLs in yeast. Here we discuss this work in the general context of genetic dissection of complex traits and QTLs.


Structure-Based Virtual Screening and In Vitro Evaluation of New Trypanosoma cruzi Cruzain Inhibitors.
Chagas disease (CD), or American trypanosomiasis, causes more than 10,000 deaths per year in the Americas. Current medical therapy for CD has low efficacy in the chronic phase of the disease and serious adverse effects; therefore, it is necessary to search for new pharmacological treatments. In this work, the ZINC15 database was filtered using the N-acylhydrazone moiety and a subsequent structure-based virtual screening was performed using the cruzain enzyme of Trypanosoma cruzi to predict new potential cruzain inhibitors. After a rational selection process, four compounds, Z2 (ZINC9873043), Z3 (ZINC9870651), Z5 (ZINC9715287), and Z6 (ZINC9861447), were chosen to evaluate their in vitro trypanocidal activity and enzyme inhibition. Compound Z5 showed the best trypanocidal activity against epimatigote (IC50 = 36.26 ± 9.9 μM) and trypomastigote (IC50 = 166.21 ± 14.5 μM and 185.1 ± 8.5 μM on NINOA and INC-5 strains, respectively) forms of Trypanosoma cruzi. In addition, Z5 showed a better inhibitory effect on Trypanosoma cruzi proteases than S1 (STK552090, 8-chloro-N-(3-morpholinopropyl)-5H-pyrimido[5,4-b]-indol-4-amine), a known cruzain inhibitor. This study encourages the use of computational tools for the rational search for trypanocidal drugs.


Primary therapy in breast cancer: what have we learned from landmark trials?
Primary anticancer therapy is currently accepted as a therapeutic option for patients with early-stage breast cancer. Its objectives are to increase the chance of achieving a conservative surgery and, similar to adjuvant chemotherapy, to reduce the risk of distant recurrence. The prognostic significance of obtaining a pathological complete response has been evaluated in several randomized clinical trials and meta-analyses. Growing evidence suggests that pathological complete response may act as a valid predictor of overall survival. Of note, a significant association between pathological complete response and outcome has especially been observed in patients with HER2-positive and triple-negative (hormonal receptors negative and HER2-negative) breast cancer. This review focuses on recent trials of neoadjuvant treatment with specific attention to HER2-negative disease.


Production and secretion of immunoglobulins in the gastrointestinal tract.
Two decades ago it was shown that the major immunoglobulin (Ig) present in human secretions is a dimeric IgA covalently bound to an epithelial glycoprotein of about 80 kD, now called the secretory component (SC). Pentameric IgM is likewise actively enriched in most exocrine fluids and is associated with SC, although not in a covalently stabilized complex. Three findings explain the selective translocation of polymeric Ig (pIg) into exocrine fluids: (1) preferential local production; (2) J-chain-expressing capacity of pIg-producing immunocytes; and (3) SC-mediated epithelial transport. Human hepatocytes lack SC and the human liver, therefore, cannot act as an efficient "IgA pump". This is in contrast to the rat liver which shows a remarkable capacity for transport of dimeric IgA from blood into the bile. The J chain of pIg and the epithelial SC represent the "lock and key" in the glandular transport of secretory IgA (SIgA) and SIgM. It has recently been shown that SC is synthesized as a transmembrane protein of about 95 kD and constitutes the actual pIg surface receptor. Complexing between ligand and receptor in the plasma membrane is followed by endocytosis. The completed SIgA and SIgM molecules are then translocated in cytoplasmic vesicles through the epithelial cell to the gland lumen along with an excess of free SC. The main function of SIgA is to exert immune exclusion; that is, by intimate cooperation with innate nonspecific defense factors it decreases penetration of soluble antigens and inhibits epithelial colonization of bacteria and viruses. Especially in selective IgA deficiency, SIgM may exert a similar protective function since its synthesis is markedly increased in the intestinal mucosa. Leakage of IgG into exocrine fluids is enhanced by mucosal irritation. Although IgG should not be considered as a SIg, it may contribute to immune exclusion. This is seen especially in the respiratory tract where IgG is less easily subjected to proteolytic degradation than in the intestinal juice. In contrast, by activating complement, IgG antibodies may at the same time be phlogistic and accelerate mucosal penetration of antigens. IgG may thus contribute to persistent immunopathology in mucosal disease. The same is true for IgE antibodies which may be carried into mucous membranes and secretions by mast cells and cause their degranulation with local histamine release. Traces of IgD may likewise be found in the secretions but without obvious biologic significance. Regulation of secretory immunity takes place both in organized lymphoepithelial structures, such as the Peyer's patches, and adjacent to the glands in the lamina propria.(ABSTRACT TRUNCATED AT 400 WORDS)


Effects of knee flexion angle and loading conditions on the end-to-end distance of the posterior cruciate ligament: a comparison of the roles of the anterolateral and posteromedial bundles.
It is commonly accepted that the anterolateral (AL) bundle of the posterior cruciate ligament (PCL) is tight in flexion and that the posteromedial (PM) bundle is tight in extension. However, a recent in vivo study showed that both bundles were tight in extension. To investigate the effects of knee flexion angle, rotational torque, and anterior/posterior translational force on the end-to-end distance between the femoral and tibial insertion sites of each bundle of the PCL. Descriptive laboratory study. Cadaveric knees (10 specimens) were mounted on a robotic system, and the relative positional data between the femur and tibia were acquired during passive flexion/extension, with an applied 5-N·m rotational torque and an applied 89-N translational force. The bony surface and PCL insertion data were acquired with a 3-dimensional scanner after gross dissection and were superimposed onto the positional data. The end-to-end distance between the 2 PCL insertion sites of the femur and tibia was measured. The end-to-end distance increased from full extension to 90° for both the AL (9.2 ± 1.8 mm; from 30.0 to 39.2 mm) and PM bundles (5.8 ± 2.2 mm; from 32.0 to 37.7 mm). With an internal rotational torque, the end-to-end distance of the PM bundle increased significantly (P < .05) at 0°, 30°, and 60° of knee flexion. Under a posterior translational force at 90° of knee flexion, the length of both bundles increased to their longest measurements (AM bundle: 40.6 ± 4.2 mm; PM bundle: 38.4 ± 3.8 mm). The end-to-end distance of the AL and PM bundles of the PCL increased in flexion, and this pattern was maintained during tests with posterior translational force. The PM bundle was more affected by the rotational torque than was the AL bundle. Both bundles of the PCL may serve a greater functional role in flexion than in extension. The PM bundle might be more important for the control of rotation than the AL bundle. Posterior translation at 90° of knee flexion could be the most stressful condition for both bundles of the PCL, which may have implications for an injury mechanism.


CA/GT microsatellite alleles within the cystic fibrosis transmembrane conductance regulator (CFTR) gene are not generated by unequal crossingover.
The gene responsible for cystic fibrosis (CF) has recently been identified, and a three-nucleotide deletion (delta F508 mutation) that results in the loss of a phenylalanine residue in the first putative ATP-binding domain of the predicted protein (CF transmembrane conductance regulator, CFTR) has been found to be the major CF mutation. Although several other mutations have been identified in the CFTR gene, most of them are very rare, making their application to genetic diagnosis difficult. While characterizing the genomic region encompassing the CF locus, we have identified three CA/GT blocks that flank exon 9 of the CF gene. One of the CA/GT blocks exhibits a highly informative variable number of dinucleotide repeats (VNDR) polymorphism. This intragenic VNDR microsatellite should, by itself, provide full information for genetic analysis in approximately 80% of CF families and will help elucidate the associations between DNA polymorphism haplotypes and specific gene mutations. Haplotype analyses of CF chromosomes with and without the delta F508 mutation suggest that the different alleles are generated by slipped-strand mispairing within the dinucleotide repeat during DNA replication, rather than by unequal crossingover within a recombination hot spot.


Effects of lipodystrophy on quality of life and depression in HIV-infected men on HAART.
The aim of the study was a prospective assessment of the possible consequences of a diagnosis of lipodystrophy on health-related quality of life (HRQL) and depressive symptomatology in HIV-seropositive men who have sex with men. A standardized physical assessment for lipodystrophy was introduced within a prospective study in April 1999. Over a 2-year follow- up, 37 HIV-seropositive men who met the criteria for lipodystrophy were longitudinally compared to 92 HIV-seropositive men without lipodystrophy and 88 HIV-seronegative men on measures of HRQL and depression. A series of questionnaires, which included the Medical Outcomes Study Short-Form 36 (SF-36) and the Center for Epidemiological Studies-Depression (CES-D), were administered to assess HRQL and depression, respectively. SF-36 scores were summarized using the mental and physical components; CES-D results were reported as both dichotomous (with or with clinical depression) and continuous scores. Neither the mental nor physical components of the SF-36 showed any significant differences between patients with lipodystrophy versus HIV-seropositive patients without lipodystrophy. Similarly, lipodystrophy status was not significantly associated with either continuous depression scores or presence of clinical depression. However, consistent with previous results, HIV-seropositive men without lipodystrophy (compared to HIV-seronegative men) reported higher scores on both components of the SF-36 scales and both categorizations of the CES-D. The results of this study suggest that lipodystrophy does not negatively affect HRQL or depression, above and beyond, the diagnosis of HIV infection, although the impact of the severity of lipodystrophy on these conditions will require further study.


Production and characterization of an anti-idiotypic single chain Fv that recognizes an anti-DNA antibody.
A well-characterized recombinant anti-idiotype to an anti-DNA antibody can be useful for studies of the regulation of anti-DNA-producing B cells. Using a hybridoma technique, a monoclonal anti-idiotypic antibody, designated O2F3, was obtained, and its scFv gene was constructed. O2F3 single chain Fv (scFv) was produced against an idiotope of a monoclonal anti-DNA antibody, 3D8, that was obtained from an autoimmune-prone mouse, MRL-lpr/lpr. Here we describe the production and in vitro characterization of the O2F3 scFv, and compare it with its parent monoclonal antibody, O2F3 IgM. To characterize O2F3 scFv and O2F3 IgM, we generated recombinant 3D8 fragments, including 3D8 scFv, 3D8 VH, and 3D8 VL, that were used as antigens in several assays. ELISA and Western blot analysis showed that both O2F3 scFv and O2F3 IgM recognized a conformational determinant formed by the association of the variable region heavy and light chains of the 3D8 antibody, suggesting that O2F3 scFv retained a similar binding pattern to its parent O2F3 antibody. The idiotope recognized by O2F3 was shown by competitive ELISA to be outside of the DNA binding site of the 3D8 antibody. This characterized O2F3 scFv could be applied for the regulation of anti-DNA antibody production and the manipulation of recombinant antibody-based proteins to which toxins, enzymes, and chemical agents can be connected.


Three-Dimensional Culture Systems for Dissecting Notch Signalling in Health and Disease.
Three-dimensional (3D) culture systems opened up new horizons in studying the biology of tissues and organs, modelling various diseases, and screening drugs. Producing accurate in vitro models increases the possibilities for studying molecular control of cell-cell and cell-microenvironment interactions in detail. The Notch signalling is linked to cell fate determination, tissue definition, and maintenance in both physiological and pathological conditions. Hence, 3D cultures provide new accessible platforms for studying activation and modulation of the Notch pathway. In this review, we provide an overview of the recent advances in different 3D culture systems, including spheroids, organoids, and "organ-on-a-chip" models, and their use in analysing the crucial role of Notch signalling in the maintenance of tissue homeostasis, pathology, and regeneration.


Prediabetes and working memory in older adults.
Insulin sensitivity, pancreatic β-cell function, fasting glucose, and 2-h post-load glucose were related to cognition in cognitively healthy nondiabetic older adults. Thirty-five adults (⩾65 years) underwent a 2-h oral glucose tolerance test and cognitive testing. Seventeen had normal glucose tolerance and 18 had intermediate hyperglycaemia or prediabetes (World Health Organization criteria). Fasting glucose and 2-h post-load glucose and oral glucose tolerance test-derived measures of β-cell function (oral disposition index) and insulin sensitivity were analysed as predictors of four cognitive domains: verbal episodic memory, verbal fluency, executive function, and working memory. The prediabetes group had significantly worse working memory performance than the normal glucose tolerance group. Controlling for age and education, decreased oral disposition index, and increased 2-h post-load glucose were significantly related to worse working memory performance. Prediabetes may worsen working memory in healthy older adults. Reduced pancreatic β-cell function should be investigated as a contributor to age-related cognitive decline.


Contamination of community water sources by potentially pathogenic vibrios following sea water inundation.
Potentially pathogenic members of the Vibrionaceae family including Vibrio cholerae and Vibrio parahemolyticus were isolated from domestic sources of drinking water in coastal villages following sea water inundation during the tsunami in Southern India. Phenotypic and genotypic studies were done to confirm the identity and detection of toxins. Vibrio-gyr (gyrase B gene) was detected in all sixteen vibrio isolates. Toxin regulating genes i.e.: ctx gene, tdh gene, and trh gene, however were not detected in any of the strains, thereby ruling out presence of toxins which could endanger human life. Other potentially pathogenic bacteria Aeromonas and Plesiomonas were also isolated from hand pumps and wells, in a few localities. There was no immediate danger in the form of an outbreak or sporadic gastroenteritis at the time of the study. Timely chlorination and restoration of potable water supply to the flood affected population by governmental and nongovernmental agencies averted waterborne gastroenteritis. Assessment of quality of water and detection of potential virulent organisms is an important public health activity following natural disasters. This work highlights the importance of screening water sources for potentially pathogenic microorganisms after natural disasters to avert outbreaks of gastroenteritis and other infectious diseases.


Discriminative stimulus properties of 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane [(+/-)DOI] in C57BL/6J mice.
The drug discrimination procedure has proven to be a valuable tool for studying the mechanism of action of psychoactive drugs. Recently, mice with targeted gene mutations have been developed that may also prove useful in evaluating the role of specific receptors in mediating the actions of drugs. We were interested in studying the effects of hallucinogens in genetically modified mice using the drug discrimination procedure. To establish the training procedures and characterize the stimulus properties of 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane [(+/-)DOI] versus saline in wild-type mice. Using a two-lever drug discrimination procedure, C57BL/6J mice were trained to discriminate (+/-)DOI (2.5 mg/kg) from saline on a VI 30-s schedule of reinforcement. The acquisition function was orderly and similar to that found previously with rats, although the training dose required for the mice was four times higher (2.5 versus 0.75 mg/kg). The dose-response relationship indicated that percent drug lever responding was dose-dependent. Two other hallucinogens, LSD and (-)DOB, substituted fully for (+/-)DOI. Mice were tested for their ability to discriminate (+/-)DOI following pretreatment with the 5-HT(2A) receptor antagonist MDL 100,907, or with 5-HT(2C) selective antagonists, SB 206,553 or SB 242,084. A dose of 0.25 mg/kg MDL 100,907 essentially completely blocked the discriminative stimulus effects of 2.5 mg/kg (+/-)DOI. Surprisingly, both SB 206,553 and SB 242,084 also attenuated the effect of 2.5 mg/kg (+/-)DOI. The effect of SB 206,553 was surmountable at 5.0 mg/kg (+/-)DOI. These data agree with the results from studies with rats indicating a prominent role for the 5-HT(2A) receptors in mediating the discriminative stimulus effects of (+/-)DOI but in addition, suggest a small but significant role for the 5-HT(2C) receptor in mice.


Proposal of an end-to-end emergency medical system.
A new comprehensive emergency system has been proposed to facilitate and computerize all the processes involved in an emergency from the initial contact to the ambulance emergency system, to finding the right and nearest available ambulance, and through to accessing a Smart Online Electronic Health Record (SOEHR). The proposed system will critically assist in pre-hospital treatments, indentify availability of the nearest available specialized hospital and communicate with the Hospital Emergency Department System (HEDS) to provide early information about the incoming patient for preparation to receive and assist.


Engineered GFP as a vital reporter in plants.
The green-fluorescent protein (GFP) of the jellyfish Aequorea victoria has recently been used as a universal reporter in a broad range of heterologous living cells and organisms. Although successful in some plant transient expression assays based on strong promoters or high copy number viral vectors, further improvement of expression efficiency and fluorescent intensity are required for GFP to be useful as a marker in intact plants. Here, we report that an extensively modified GFP is a versatile and sensitive reporter in a variety of living plant cells and in transgenic plants. We show that a re-engineered GFP gene sequence, with the favored codons of highly expressed human proteins, gives 20-fold higher GFP expression in maize leaf cells than the original jellyfish GFP sequence. When combined with a mutation in the chromophore, the replacement of the serine at position 65 with a threonine, the new GFP sequence gives more than 100-fold brighter fluorescent signals upon excitation with 490 nm (blue) light, and swifter chromophore formation. We also show that this modified GFP has a broad use in various transient expression systems, and allows the easy detection of weak promoter activity, visualization of protein targeting into the nucleus and various plastids, and analysis of signal transduction pathways in living single cells and in transgenic plants. The modified GFP is a simple and economical new tool for the direct visualization of promoter activities with a broad range of strength and cell specificity. It can be used to measure dynamic responses of signal transduction pathways, transfection efficiency, and subcellular localization of chimeric proteins, and should be suitable for many other applications in genetically modified living cells and tissues of higher plants. The data also suggest that the codon usage effect might be universal, allowing the design of recombinant proteins with high expression efficiency in evolutionarily distant species such as humans and maize.


Interaction of porphyran with a hydrophobic surface and stabilization of liposomes.
Three porphyran preparations with high emulsifying ability and varying molecular mass, 3,6-anhydrogalactose content, and sulfate content without any protenaceous component were prepared from dried nori processed from Porphyra yezoensis, a red alga. Each of these preparations was applied to demonstrate adsorption or binding to the surface of oil droplets. The decrease in porphyran concentration of the aqueous phase of O/W emulsions prepared with porphyran and with toluidine blue (TB)-porphyran complex formed by adding TB to the O/W emulsions indicated ready adsorption to the surface of oil droplets. The decrease in zeta-potential of the O/W emulsions suggested that the sulfate groups of the adsorbed porphyran were oriented toward the external aqueous phase. A biomolecular interaction analysis exhibited rapid binding of porphyran to C16-alkane, probably through 3,6-anhydrogalactose. Porphyran-coated liposomes were tolerant to digestion with phospholipase D. The increased molecular weight of the porphyran preparations had an increased effect on these characteristics. The results of this study demonstrate that the emulsifying ability of porphyran is derived from the adequate adsorption to the surface of oil droplets and that porphyran could be effectively applied to stabilize liposomes.


The role of dopamine and serotonin in conditioned food aversion learning in the honeybee.
For most animals, eating entails the risk of being poisoned. Learning how to identify foods with toxins is an important mechanism that reduces the risk of poisoning. While conditioned food aversions have been studied in vertebrates for over 50 years, the neural circuits underlying this form of learning have been difficult to elucidate because of their complexity. Insects, such as fruit flies and honeybees, are important models for the study of the neural mechanisms of learning and memory, but conditioned food aversions have not yet been reported from either species. My collaborators and I recently established that the honeybee has the ability to learn to avoid odors associated with toxins in food using two independent neural pathways. In these experiments, we found that honeybees can learn to associate scents with toxins that they can pre-ingestively detect using their proboscis. This form of learning is primarily mediated by the neurotransmitter, dopamine. We also found a second mechanism: bees can learn to avoid odors associated with the malaise caused by ingesting toxins. This form of learning is mediated by serotonin. Our data are the first to show that two different mechanisms account for conditioned food aversions in insects.


Synthesis and structure activity relationship of tetrahydroisoquinoline-based potentiators of GluN2C and GluN2D containing N-methyl-D-aspartate receptors.
We describe here the synthesis and evaluation of a series of tetrahydroisoquinolines that show subunit-selective potentiation of NMDA receptors containing the GluN2C or GluN2D subunits. Bischler-Napieralski conditions were employed in the key step for the conversion of acyclic amides to the corresponding tetrahydroisoquinoline-containing analogs. Compounds were evaluated using both two-electrode voltage clamp recordings from Xenopus laevis oocytes and imaging of mammalian BHK cells loaded with Ca(2+)-sensitive dyes. The most potent analogues had EC50 values of 300 nM and showed over 2-fold potentiation of the response to maximally effective concentrations of glutamate and glycine but had no effect on responses from NMDA receptors containing the GluN2A or GluN2B subunits AMPA, kainate, and GABA or glycine receptors or a variety of other potential targets. These compounds represent a potent class of small molecule subunit-selective potentiators of NMDA receptors.


[Molecular study of two novel RHD alleles and pedigree analysis].
To study the segregation of two novel RHD alleles in Chinese pedigrees. The Rh antigens of the samples were identified by using monoclonal antibodies. The 10 exons of the RHD gene for the 2 probands and their family members were amplified separately and sequenced. The parents of proband 2 were analyzed by sequence specific primer-polymerase chain reaction (SSP-PCR). The two probands were RhD negative and the RHD was D/d type. After alignment with the nucleotide sequence in GenBank, a deletion of nucleotide C at position 78 in exon 1 of proband 1 was detected, and her sister also had the deletion, which was confirmed by sequencing. The sequencing results of proband 2 showed a 10 nucleotide deletion in exon 8 as well as a RHD 520 G to A mutation in exon 4. The results of SSP-PCR and sequencing showed that the proband's mother also carried RHD 520 G to A and RHD 1080 del 10 mutation, which was transmitted to proband 2. The sequences of the novel alleles have been submitted to GenBank (accession No. GQ477180 and GU362076). The two novel RHD alleles, RHD 78delC and RHD 520 G to A+1080 del 10, were both pseudo genes and stably transmitted.


Immunotoxicity of heavy metals in relation to Great Lakes.
Heavy metals including mercury, lead, and cadmium are present throughout the ecosystem and are detectable in small amounts in the Great Lakes water and fish. The main route of exposure of humans to these metals is via the ingestion of contaminated food, especially fish. Extensive experimental investigations indicated that heavy metals alter a number of parameters of the host's immune system and lead to increased susceptibility to infections, autoimmune diseases, and allergic manifestations. The existing limited epidemiologic data and data derived from in vitro systems in which human peripheral blood leukocytes were used suggested that the human immune system may also be at increased risk following exposure to these metals. The magnitude of the risk that the presence of such metals in the Great Lakes may pose to the human immune system, and consequently to their health, is not known. In this review, the available data with respect to potential adverse effects of heavy metals on the immune system of humans and experimental animals are discussed, and additional data requirements are suggested.


BKB-SIN and ANL predict perceived communication ability in cochlear implant users.
Cochlear implant (CI) users typically report that speech recognition becomes substantially more difficult in the presence of background noise. This perception is consistent with objective measures of speech recognition showing that CI users require more favorable signal-to-noise ratios than normal-hearing (NH) listeners to achieve equivalent speech recognition. However, recent research in hearing aid users suggests that noise tolerance or the "willingness to listen in noise" may also influence perceived communication ability. Therefore, the purpose of this study was to evaluate the extent to which speech recognition in noise and noise tolerance predict perceived communication ability among adult CI users. Twenty adult CI users were evaluated on an objective test of speech recognition in noise (Bamford-Kowal-Bench Sentences in Noise [BKB-SIN] test) and a measure of noise tolerance (Acceptable Noise Level [ANL] test) and completed a self-report measure of communication difficulty (Abbreviated Profile of Hearing Aid Benefit [APHAB]). Relationships between BKB-SIN scores, ANL scores, and aided APHAB scores for the ease of communication, background noise, reverberation, and Global scales were assessed. In addition, BKB-SIN scores and ANL scores for the CI users were compared with scores for a control group of 23 NH listeners. CI users demonstrated substantially poorer BKB-SIN scores than NH listeners, as expected; however, their ANL scores were similar to those for NH listeners. BKB-SIN scores and ANL scores were not systematically related to one another. Each measure accounted for more than one third of the variance in CI users' aided APHAB Global scores; together, the two measures accounted for 72% of that variance. Both speech recognition in noise and noise tolerance are strongly associated with CI users' self-perceived communication ability. The two measures seem to reflect different factors that influence an individual's communication experience; thus, both may provide useful clinical information. The establishment of formal criteria for BKB-SIN scores and ANL scores that are predictive of excessive communication difficulty may help to identify CI users who could benefit from additional audiologic rehabilitation.


MUSIC Speciation of γ-Al2O3 at the Solid Liquid Interface: How DFT Calculations Can Help with Amorphous and Poorly Crystalline Materials.
Transition aluminum oxides, such as γ-Al2O3 or alumina, are widely used in many different technical applications that rely on the surface reactivity of this material at the solid liquid interface. The speciation of surface sites of this material confronts several obstacles. On the one hand, alumina is a poorly crystalline oxide, thus allowing for a limited amount of empirical structural information for an important number of surface sites with different trends in reactivity and, on the other hand, it is a metastable material. In this work, we show several ways in which the multisite complexation model, combined with atomistic information from density functional theory and ab initio molecular dynamics (AIMD), can manage to perform speciation calculations of γ-Al2O3 surface sites at the solid liquid interface. Although the results are in good qualitative agreement with experimental titration curves, and they can serve as a guide for the interpretation of the reactivity of this material at the initial stages of an impregnation experiment and chemical weathering phenomena, this work highlights the need of more complex AIMD simulations to accurately model these phenomena in γ-Al2O3 surface/liquid interfaces.


[Magnetic resonance myelography with a fast-spin-echo sequence].
A major application of Magnetic Resonance Imaging (MRI) consists in the "myelographic effect", characterized by high signal intensity of the CSF, which permits to visualize spinal subarachnoid spaces and nerve root sheaths. Such new ultrafast pulse sequences as fast spin echo (FSE) further enhance CSF high signal intensity. In addition, suppressing background with the fat suppression technique and with maximum intensity projection (MIP), a particular algorithm already used in MR angiography, FSE sequences yield 3D myelography-like images. Our study was aimed at assessing the clinical role and the reliability of this technique. Our preliminary experience is based on 21 patients with pain in the lower back and legs and on 5 healthy volunteers; all the myelography-like images were obtained with a 1.5-T MR system (Signa, General Electric). The parameters for FSE myelography-like images included: TR = 6000, TE = 200, ETL = 32, thickness = 3 mm, matrix = 256 x 224, Nex = 4, FOV = 20 cm, fat presaturation, no phase wrap, acquisition time = 2 min 58 s. MR myelography sequences were acquired after a standard FSE T1/T2-weighted exam. MR-myelography results confirmed the diagnosis of disk herniation made on conventional T1/T2-weighted images, especially in thecal sac compression and/or amputation of nerve root sheaths. MR myelography appeared to be especially useful in migrated disk herniation or in abnormal origin of nerve roots. Moreover, it allowed full and panoramic visualization of the subarachnoid spaces and excellent definition of the thecal sac, nerve roots and nerve root sheaths. In one patient only movement artifacts affected image quality. To conclude, even though it fails to yield any functional information, MR myelography appears to be an easy, rapid and noninvasive support to conventional MRI of the lumbar spine, to replace conventional myelography.


Distribution of PACAP in the brain of the cartilaginous fish torpedo marmorata.
In this article, we investigated the distribution of pituitary adenylate cyclase-activating polypeptide (PACAP) and its mRNA by immunohistochemistry, in situ hybridization, and RT-PCR techniques, in the central nervous system of the elasmobranch Torpedo marmorata. RT-PCR analysis showed that the CNS of T. marmorata expresses a messenger encoding PACAP. The immunohistochemistry and in situ hybridization patterns were partly overlapping, with a major expression in the hypothalamo-pituitary region and, surprisingly, in the saccus vasculosus. Our results show that, in T. marmorata, PACAP is synthesized and widely distributed in the CNS, suggesting an as yet unidentified role for this peptide in elasmobranch brain physiology.


Dramatic long-term restoration of an oak woodland due to multiple, sustained management treatments.
We measured 34 years of plant community change in a degraded oak woodland undergoing ecological management. Management included regular prescribed fire, control of white-tailed deer populations, repeated sowing of a diverse seed mix, and removal of invasive plants. We tracked change with several conservation metrics. Time series analysis showed no significant changes over time in either plant species richness or the Shannon-Weiner diversity index. Floristic Quality Assessment measures-the Floristic Quality Index (FQI), Cover-weighted FQI, and the Mean Coefficient of Conservatism (Mean C)-all increased dramatically over time, such that their values now surpass those of the highest quality representative of this habitat in the region. Cover-weighted FQI had the added benefit of being quick to respond (negatively and positively) to short-term management changes during the study. This sensitivity highlights its utility for adaptive management, enabling timely, data-driven changes to ongoing management regimes. Plant community composition showed striking changes during the study period, as species of high conservation value replaced weedier species. As a group, conservative woodland species are notoriously slow to recover from degradation, making this flora's recovery particularly notable. A mid-study cessation of management immediately stalled the woodland's recovery according to Floristic Quality metrics, but the restoration quickly returned to its positive trajectory with the resumption of management treatments. These results illustrate that impressive plant biodiversity restoration can be achieved, even in highly degraded contemporary oak ecosystems, if ecological management is comprehensive and if it is sustained over time.


Thumb base involvement in symptomatic hand osteoarthritis is associated with more pain and functional disability.
To assess the impact of different subsets of symptomatic hand osteoarthritis (OA) on pain and disability. From 308 patients with hand OA a group with carpometacarpal joint (CMCJ) symptoms only (group I, n=20) was identified as well as groups with symptoms at the interphalangeal joints (IPJs) only (group II, n=138), and symptoms at both sites (group III, n=150). Hand pain and function, assessed with the AUSCAN, were compared between groups using linear mixed models. Radiological OA was assessed using the Kellgren-Lawrence grading scale. Mean (SD) AUSCAN scores for groups I, II and III were 23.1 (11.7), 18.3 (11.9) and 26.4 (12.5), respectively. After adjustment for age, gender, body mass index, family effects and number of symptomatic hand joints, significant differences in AUSCAN scores of 7.4 (95% CI 1.8 to 13.0) between groups I and II, and 5.7 (95% CI 2.7 to 8.6) between groups II and III were found. AUSCAN scores were 5.8 (95% CI 3.1 to 8.6) higher for patients with versus patients without CMCJ symptoms. Kellgren-Lawrence scores did not differ between groups. In symptomatic hand OA, CMCJ OA contributes more to pain and disability than IPJ OA. Hence, treatment of CMCJ OA should be emphasised, even if it coincides with IPJ OA.


Ultrasound-guided evaluation of the local anesthetic spread parameters required for a rapid surgical popliteal sciatic nerve block.
The ideal spread of local anesthetic (LA) solution around the sciatic nerve during a popliteal block remains unclear. We tested the hypothesis that a circumferential spread of LA and/or intraneural injection could lead to rapid surgical block. Patients (n = 100) scheduled for foot or ankle surgery underwent popliteal sciatic nerve block using nerve stimulation according to Borgeat's technique and injection of ropivacaine (0.5 mL/kg). Sensory and motor blockades were assessed on the tibial nerve (TN) and common peroneal nerve (CPN) at 5, 15, and 30 mins after completion of the block and in the recovery room. A successful block was defined as a complete sensory block in TN and CPN. Changes in cross-sectional and longitudinal surfaces and diameters and the characteristics of LA spread around the nerve were noted using ultrasound. A suspected intraneural injection was defined as a 15% increase in the surface area or anteroposterior diameter of the nerve. Patients were followed up on days 1 and 7 after surgery. Successful block was noted in 57% of patients at 30 mins and in 88% of patients in the recovery room. A circumferential spread of LA occurred in 47% of patients and 53% had noncircumferential spread. Complete sensory block was significantly higher in the group that had a circumferential spread (73% vs 43%, P = 0.035) only at 30 mins. In the postoperative care unit, there was no difference among the groups. Separated circumferential spreads around TN and CPN were noted in 12% of patients. All of these patients had a complete sensory and motor blockade at 15 mins. Concerning intraneural injection, only the change in the anteroposterior diameter on a 6-cm length of nerve was associated with a higher success and faster onset block at 5 (P = 0.008), 15 (P = 0.02), and 30 (P = 0.05) mins. There were no clinically detectable nerve injuries at follow-up. For popliteal sciatic nerve block, circumferential spread of LA, and separation of the nerve into its 2 components are associated with rapid surgical block.


Care of substance-exposed infants: the current state of practice in Canadian hospitals.
Many practices related to care of infants exposed to drugs and alcohol during th e prenatal period have been developed on an anecdotal basis. There are few available research studies to validate these interventions. This study describes current practices used in daily care, discharge planning, and community support for drug- and alcohol-exposed infants, their families, and alternate caregivers in one country. A survey of Canadian hospitals with annual birth rates of greater than 500 was conducted with a 51% return rate. Variations were noted in the perception of the importance of the issue of perinatal substance use, nursery care, discharge planning, and community support. Nurses are encouraged to consider these results within the context of their own communities to evaluate and develop standards of practice for substance-exposed infants and their families and caregivers.


Gradients of polyamines and their biosynthetic enzymes in coleoptiles and roots of corn.
The distribution of diamines, polyamines, and their biosynthetic enzymes arginine decarboxylase and ornithine decarboxylase in roots and coleoptiles of corn (Zea mays var Golden Cross Bantam) seedlings have been determined. Putrescine content, expressed on either a fresh weight or protein basis, increases from the tip to the base in both roots and coleoptiles. In roots, this gradient is paralleled by an activity gradient of arginine and ornithine decarboxylases. Spermidine is distributed equally along the length of coleoptiles; in roots, this is true only on a protein basis. Free spermine is detectable only in the root tip, but a bound form is present throughout the root and coleoptile. The results are compared with gradients in protein and DNA content and discussed in relation to the possible cellular roles of polyamines.


Increasing rice productivity and yield by manipulation of starch synthesis.
Plant productivity and yield are dependent on source-sink relationships, i.e. the capacity of source leaves to fix CO2 and the capacity of developing sink tissues and organs to assimilate and convert this fixed carbon into dry matter. Studies from our laboratories as well as others have demonstrated that rice productivity and yield are mainly sink-limited during its development because of limited capacity to utilize the initial photosynthetic product (triose phosphate). This limitation in triose phosphate utilization, evident at both the vegetative and reproductive stages of rice development, may be associated with limited capacity for carbohydrate synthesis in rice leaves (which are poor accumulators of starch) or feedback due to limited sink strength of developing seeds. Strategies in improving triose phosphate utilization by enhancing starch production in leaves and developing seeds by the expression of engineered genes for ADP glucose pyrophosphorylase, a key regulatory enzyme of starch biosynthesis, are discussed.


Bilateral small radial forearm flaps for the reconstruction of buccal mucosa after surgical release of submucosal fibrosis: a new, reliable approach.
Oral submucous fibrosis is a collagen disorder affecting the submucosal layer and often severely limiting mouth opening. Previous surgical treatments have been disappointing. This article introduces a new surgical approach: reconstructing the bilateral buccal mucosa with two small radial forearm flaps. The surgical method includes the complete surgical release of fibrotic buccal mucosa and, if necessary, a bilateral coronoidectomy and temporalis muscle myotomy. From 1997 to 1999, 15 patients with moderate-to-severe trismus received reconstructive surgery, for a total of 30 small radial forearm flaps after surgical release. The flap size was between 1.5 x 5 and 2.5 x 7 cm. All donor sites were directly closed, and all flaps survived completely, except for one with partial necrosis. Six flaps required minor revisions because of size redundancy. Two patients developed buccal cancer in the area of reconstruction. At an average of 12 months' follow-up, the inter-incisal distance averaged 33 mm, an increase of 17 mm compared with the preoperative value. The donor-site morbidity was minimal, except in one heavy smoker who developed dry gangrene of his fingertips. The use of two small free forearm flaps for buccal mucosa reconstruction allows more radical release of fibrotic tissue. Coronoidectomy and temporal muscle myotomy further contribute to the effect of trismus release. The combined effects of this approach have consistently given good results. An aggressive approach toward surgical treatment of this precancerous lesion also facilitates the detection of cancer at an early stage.


Methods for locating the tibio-femoral contact pathway in total knee replacements using marker-based gait analysis and standard radiography.
The purpose of this study was to develop and test techniques for tracking the path of contact between the tibial and femoral total knee replacement components during level over-ground walking. The tibio-femoral path of contact could be an indicator of the in vivo performance of a total knee replacement as an estimator of areas of contact between the implant components. A longer contact path, indicative of more sliding between the implant components during walking, could indicate an implant at risk for increased wear. In addition, the tibio-femoral contact path determines the position and length of the muscle and ligament lever arms about the knee, and can subsequently influence knee contact force calculations. Two methods were developed to predict the tibio-femoral contact pathways for total knee replacement devices. Both methods used patient-specific knee kinematics obtained during gait analysis, standard radiographs obtained during clinical follow-ups, and point-clouds of the tibial and femoral bearing surfaces. The validity of the techniques was evaluated with knee wear simulator tests and comparisons to wear scars on postmortem retrieved tibial components. The average total anterior-posterior distance covered by the contact path for ten patients implanted with a total knee replacement was 29.01 mm on the lateral side, and 21.80 mm on the medial side. Both methods for predicting the tibiofemoral contact pathways yielded similar results, and fell within the wear scars of simulator-tested and postmortem retrieved implants. The methods for predicting the tibio-femoral contact pathway using marker-based gait analysis and standard clinical radiographs are computationally simple, and reliably predict contact path characteristics as evaluated against wear scars from knee wear simulator tests and postmortem retrieved implants.


Acacia sieberiana (Fabaceae) attenuates paracetamol and Bile Duct Ligation-Induced hepatotoxicity via modulation of biochemical and oxidative stress biomarkers.
Background: The plant Acacia sieberiana (Fabaceae) is traditionally used to manage hepatitis. This research work aims to investigate the hepatoprotective effectiveness of root bark extract of Acacia sieberiana (ASE) against paracetamol (PCM) and bile duct ligation (BDL)-induced hepatotoxicity. The phytochemical and median lethal dose (LD50) investigations were conducted. The rats were pre-treated with the ASE (250, 750, and 1,500 mg/kg) once daily via oral route for 7 consecutive days. On the 8th day, liver injury was initiated by PCM administration (2 g/kg). Similarly, in the BDL-induced liver injury, the animals were administered ASE (125, 250, and 380 mg/kg) intraperitoneally for 7 consecutive days. After 24 h, blood samples and hepatic tissues were obtained for biochemical and histopathological investigations. Results: Phytocomponents determination revealed glycosides, triterpenes, glycosides, saponins, tannins, flavonoids and alkaloids. The oral and intraperitoneal LD50 values of the ASE were >5,000 and 1,300 mg/kg, respectively. The ASE efficiently (p < 0.05) decreased the alanine transaminase (ALT) and aspartate transaminase (AST) levels and elevated the albumin and total protein (TP) levels. The direct bilirubin effectively (p < 0.05) decreased at 750 mg/kg. Besides, the extract efficiently elevated the glutathione peroxidase (GPx), superoxide dismutase (SOD), and catalase (CAT) in relation to the PCM hepatotoxic group. Also, the malondialdehyde (MDA) concentration was reduced by the ASE. Meanwhile, in the BDL-induced liver injury, the ASE remarkably (p < 0.05) declined the AST, ALP, bilirubin,and MDA. Besides, there was effective (p < 0.05) elevation in SOD, GPx and CAT in the ASE-treated groups. The morphology of liver tissue was preserved at 125 and 250 mg/kg ASE groups from BDL-induced necrosis and vascular congestion. Conclusion: The study shows that the ASE has hepatoprotective actions against liver damage by possible modulation of biochemical and oxidative stress biomarkers.


[Analysis of antimicrobial resistance of Streptococcus pneumoniae with restriction fragment length polymorphism of pbp2b gene and pulsed-field gel electrophoresis profiles among children].
Streptococcus pneumoniae is a common cause of potentially life-threatening infections such as meningitis, bacteraemia, pneumonia worldwide, for which children of preschool age are at particularly high risk. Since the late 1970s and 1980s, antibiotic resistance among pneumococci has become an emerging problem. Several multidrug-resistant clones have rapidly spread throughout the world. (1) To investigate the prevalence of penicillin and other antibiotics nonsusceptibility among pneumococci. (2) To analyze the correlation of pbp2b amplicon profiles with penicillin resistance. (3) To serotype 31 isolates of penicillin-resistant pneumococci by latex agglutination. (4) To analyze the chromosomal relatedness of serotype 23F and 6 isolates of penicillin-resistant pneumococci by using pulsed-field gel electrophoresis (PFGE) and characterize these isolates in molecular epidemiology. (1) Susceptibility was determined by using broth microdilution, E-test, and K-B disk. (2) The correlation of pbp2b amplicon profiles with penicillin resistance was assessed by restriction fragment length polymorphism (RFLP). (3) Serotyping of penicillin-resistant pneumococcal isolates was performed by using latex agglutination. (4) The properties of serotype 23F and 6 isolates of penicillin-resistant pneumococci were assessed by PFGE. S. pneumoniae with increased nonsusceptibility (including intermediate strains and resistant strains) to penicillin G was 9.9% in 1997, 12.6% in 1998, 14.6% in 2000; to cefuroxime 4.2%, 1.5%, 8.2%; to cefotaxime 0.0%, 1.7%, 1.0% respectively. There were no statistically significant differences (P > 0.05). While resistance to erythromycin, trimethoprim-sulfamethoxazole and chloramphenicol increased significantly from 76.8% in 1997 to 87.4% in 2000, from 74.7% to 88.3%, and from 22.6% to 40.8%, respectively (P < 0.05). RFLP analysis of pneumococcal pbp2b-specific amplicons was effective for screening penicillin resistance. Of the 31 strains of penicillin-resistant pneumococci (MICs 0.12 - 2.0 micro g/ml) studied, 6 (19.4%) strains (MICs 0.12 - 0.19 micro g/ml) were serotype 23F and 3 (9.7%) strains (MICs 0.5 - 1.5 micro g/ml) were serotype 6. There were nearly identical susceptibility to antibiotics and identical PFGE patterns in the former, and there were different susceptibility to antibiotics and different PFGE patterns in the latter. Three serotype 6 strains had different susceptibility to antibiotics and different PFGE patterns, which suggested that those strains may be scattered. Generally beta-lactams retained their activity against S. pneumoniae in Beijing. Resistance to erythromycin, trimethoprim-sulfamethoxazole, and chloramphenicol increased drastically. RFLP analysis of pneumococcal pbp2b-specific amplicons was effective for screening penicillin resistance. In 6 strains of serotype 23 F there were nearly identical susceptibility to antibiotics and identical PFGE patterns, which suggested the probability that there was a spread of serotype 23F isolates with low-level penicillin resistance in local area.


Integrated mRNA-microRNA profiling of human NK cell differentiation identifies MiR-583 as a negative regulator of IL2Rγ expression.
Natural killer (NK) cells are innate immune effector cells that protect against cancer and some viral infections. Until recently, most studies have investigated the molecular signatures of human or mouse NK cells to identify genes that are specifically expressed during NK cell development. However, the mechanism regulating NK cell development remains unclear. Here, we report a regulatory network of potential interactions during in vitro differentiation of human NK cells, identified using genome-wide mRNA and miRNA databases through hierarchical clustering analysis, gene ontology analysis and a miRNA target prediction program. The microRNA (miR)-583, which demonstrated the largest ratio change in mature NK cells, was highly correlated with IL2 receptor gamma (IL2Rγ) expression. The overexpression of miR-583 had an inhibitory effect on NK cell differentiation. In a reporter assay, the suppressive effect of miR-583 was ablated by mutating the putative miR-583 binding site of the IL2Rγ 3' UTR. Therefore, we show that miR-583 acts as a negative regulator of NK cell differentiation by silencing IL2Rγ. Additionally, we provide a comprehensive database of genome-wide mRNA and miRNA expression during human NK cell differentiation, offering a better understanding of basic human NK cell biology for the application of human NK cells in immunotherapy.


Cortical cerebral microinfarcts on 3T MRI: A novel marker of cerebrovascular disease.
We examined the risk factors of cortical cerebral microinfarcts (CMIs) on 3T MRI and their association with cognitive impairment. Participants (aged 60 years and older) from the multiethnic Epidemiology of Dementia In Singapore Study underwent detailed neuropsychological testing and 3T brain MRI. Cortical CMIs were graded using a previously validated protocol. Cognitive impairment was categorized into cognitive impairment, no dementia (CIND)-mild, CIND-moderate, and dementia. Cognitive function was summarized as composite and domain-specific z scores. Among 861 participants, 54 (6.3%) had ≥1 cortical CMI. In multivariate-adjusted models, the risk factors of cortical CMIs were increasing age, Malay ethnicity, hypertension, diabetes, history of stroke, and markers of both large (cortical infarcts and intracranial stenosis) and small (lacunar infarcts, white matter hyperintensities, cerebral microbleeds) vessel disease. Presence of cortical CMIs was associated with CIND-moderate (odds ratio: 3.12; 95% confidence interval [CI]: 1.18-8.58), dementia (odds ratio: 16.92; 95% CI: 3.37-85.05), and poorer cognitive function (mean difference in composite z score: -0.42; 95% CI: -0.62 to -0.21). Additional adjustments for vascular risk factors and other MRI markers did not alter these associations. Cortical CMIs are a novel MRI marker of cerebrovascular disease and are independently associated with cognitive impairment and dementia. These findings provide new insights into the burden of cerebrovascular disease in cognitive impairment. Future research is needed to establish the additional etiologic and prognostic significance of cortical CMIs.


Stigmatizing attitudes of tertiary hospital physicians towards people with mental disorders in Saudi Arabia.
To measure stigma among doctors in one of the major specialized hospitals in Saudi Arabia. This cross-sectional study was conducted at King Abdullah Medical City, Jeddah, Saudi Arabia between October and November 2018. Eighty physicians were recruited in this study. The participants administered the sociodemographic questionnaire as well as the Mental Illness Clinicians' Attitude 4th version (MICA 4). The overall MICA score ranged between 31 and 61 with a mean±SD of 45.75±7.54. The highest reported score was among outpatient physicians (51.33±6.66), while the lowest score was among consultants/assistant consultants (43.17±7.82). The results of this study showed a relatively high MICA-4 score that could indicate a high stigmatizing attitude among physicians toward patients with mental illnesses compared with the other MICA-4 studies. Thus, training workshops could improve the attitudes in the short-term.


Isolation, characterization and pharmacological activity of Magydaris pastinacea (Lam) Paol. glucosides.
The glycoside fraction of fresh rhizomes from Magydaris pastinacea (Lam.) Paol. was separated from the alcoholic extract using the method of Kobayashi et al. The fraction was found to have six main constituents, the most abundant of which had previously been isolated and identified as 7-O-beta-D-glucopyranosyl-8-(2',3'-dihydroxy-3-methyl-butylcoumarin++ +). The present paper describes the separation and characterization of the other constituents, all with coumarin or furancoumarin structures. Pharmacological experiments to test these compounds as platelet antiaggregants are also reported.


[The varicocele today (anatomic and physiopathologic aspects)].
Primary varicocele is a very important clinical form in establishing of male infertility. It's origin is'nt ever caused by anatomical alterations of internal spermatic vein. Clinical picture is always joined with histopathological alterations of various degree, which, partial and in concomitance with hemodynamic disorders, explains the arising of spermatogenesis alterations.


[Treatment of hepatic veno-occlusive disease after bone marrow transplantation with recombinant human tissue plasminogen activator (rh-tPA)].
Ten children were treated with recombinant human tissue plasminogen activator (rh-tPA) for severe hepatic veno-occlusive disease (VOD) that developed after bone marrow transplantation. Treatment with rh-tPA was begun a median of 22 days (range; 13-127 days) after transplantation. Seven of 9 (78%) evaluable patients had complete resolution of their VOD. Four patients had hemorrhagic complications, and 2 of them died because of pulmonary hemorrhage and subdural hemorrhage, respectively. Although rh-tPA seems to be an effective therapy for established VOD, further studies will be necessary to determine its safety as well as the optimal dosing regimen.


Senator Charles H. Percy: great friend of the elderly.
As can be seen from this quick analysis. Senator Charles Percy has certainly left his mark in the United States Senate and on the nation as well. His accomplishments have been many. He has used his truly incredible intellect and energy level for the benefit of all Americans. All American citizens owe Senator Percy a debt of gratitude for his years of public service. He is proof that the American system of government endures and that the best and the brightest still look to public service. It says a great deal about the man that he has spent so much of his time dealing with the problems of the aged, disabled, and infirm. He has been the voice of the voiceless and the conscience of the country demanding the recognition and redress of grievances, an end to the neglect, violence and discrimination perpetrated against the weakest members of society. CARING is proud to salute this truly great American.


Cornus officinalis Methanol Extract Upregulates Melanogenesis in Melan-a Cells.
Cornus officinalis is widely distributed in Korea, and its fruit has been used to make as herbal drug for traditional medicine in Korea, Japan, and China because of its tonic, analgesic, and diuretic properties. However, the effects of C. officinalis methanol extract (COME) on melanogenesis remain poorly understood. We evaluated the melanogenic capability of COME in melan-a cells, which are immortalized mouse melanocytes. COME increased melanin synthesis in a dose-dependent manner. Treatment with 12.5 μg/mL of COME significantly increased melanin content by 36.1% (p < 0.001) to a level even higher than that (31.6%) of 3-isobutyl-1-methyl-xanthine, a well-known pigmentation agent. COME also upregulated tyrosinase activity and its messenger RNA and protein expression. In addition, COME upregulated the expression of tyrosinase-related proteins 1 and 2 and microphthalmia-associated transcription factor-M messenger RNA expression. These results imply that COME may be appropriate for development as a natural product to treat hair graying.


Immature oxidative stress management as a unifying principle in the pathogenesis of necrotizing enterocolitis: insights from an agent-based model.
Necrotizing enterocolitis (NEC) is a complex disease involving prematurity, enteral feeding, and bacterial effects. We propose that the underlying initial condition in its pathogenesis is reduced ability of the neonatal gut epithelial cells (NGECs) to clear oxidative stress (OS), and that when such a NGEC population is exposed to enteral feeding, the increased metabolic OS tips the population toward apoptosis, inflammation, bacterial activation, and eventual necrosis. The multi-factorial complexity of NEC requires characterization with computational modeling, and herein, we used an agent-based model (ABM) to instantiate and examine our unifying hypothesis of the pathogenesis of NEC. An ABM of the neonatal gut was created with NGEC computational agents incorporating rules for pathways for OS, p53, tight junctions, Toll-like receptor (TLR)-4, nitric oxide, and nuclear factor-kappa beta (NF-κB). The modeled bacteria activated TLR-4 on contact with NGECs. Simulations included parameter sweeps of OS response, response to feeding, addition of bacteria, and alterations in gut mucus production. The ABM reproduced baseline cellular respiration and clearance of OS. Reduction in OS clearance consistent with clinical NEC led to senescence, apoptosis, or inflammation, with disruption of tight junctions, but rarely to NGEC necrosis. An additional "hit" of bacteria activating TLR-4 potentiated a shift to NGEC necrosis across the entire population. The mucus layer was modeled to limit bacterial-NGEC interactions and reduce this effect, but concomitant apoptosis in the goblet cell population reduced the efficacy of the mucus layer and limited its protective effect in simulated experiments. This finding suggests a means by which increased apoptosis at the cellular population level can lead to a transition to the necrosis outcome. Our ABM incorporates known components of NEC and demonstrates that impaired OS management can lead to apoptosis and inflammation of NGECs, rendering the system susceptible to an additional insult involving regionalized mucus barrier failure and TLR-4 activation, which potentiates the necrosis outcome. This type of integrative dynamic knowledge representation can be a useful adjunct to help guide and contextualize research.


Structure and stability of alkane-linked DNA hairpin conjugates.
The synthesis and properties of three related families of alkane-linked DNA hairpins are reported. The first possesses a dodecane linker (C12) and 2-8 AT base pairs. The second possesses six AT base pairs and straight chain alkane linkers having 8-16 methylenes. The third has three alkane linkers of different length and a constant six base-pair stem with alternating A-T bases and a single TT step. The spectroscopic properties (UV, CD, and (1)H NMR) and molecular modeling are consistent with the formation of base-paired B-DNA structures for all hairpins having four or more AT base pairs. The thermal stability of hairpins having a C12 linker is greater than that of the commonly used hexa(ethylene glycol) linker but less than that of the stilbenediether linker having the same AT base-pair domain. Hairpin stability is related to both hydrophobic interactions between the linker and the adjacent base pair (stilbene > alkane > glycol) and the overall length of the linker. The stability of the alkane-linked hairpins having six AT base pairs is greater for a tetradecane linker than for either shorter or longer linkers. The good thermal stability of alkane-linked hairpins and absence of a chromophore which absorbs in the UV region makes them well-suited for studies of the electronic spectra and photochemistry of short hairpins having variable base-pair sequences.


A comparison of different electrostatic potentials on prediction accuracy in CoMFA and CoMSIA studies.
Computational chemistry is playing an increasingly important role in drug design and discovery, structural biology, and quantitative structure-activity relationship (QSAR) studies. For QSAR work, selecting an appropriate and accurate method to assign the electrostatic potentials of each atom in a molecule is a critical first step. So far several commonly used methods are available to assign charges. However, no systematic comparison of the effects of electrostatic potentials on QSAR quality has been made. In this study, twelve semi-empirical and empirical charge-assigning methods, AM1, AM1-BCC, CFF, Del-Re, Formal, Gasteiger, Gasteiger-Hückel, Hückel, MMFF, PRODRG, Pullman, and VC2003 charges, have been compared for their performances in CoMFA and CoMSIA modeling using several standard datasets. Some charge assignment models, such as Del-Re, PRODRG, and Pullman, are limited to specific atom and bond types, and, therefore, were excluded from this study. Among the remaining nine methods, the Gasteiger-Hückel charge, though commonly used, performed poorly in prediction accuracy. The AM1-BCC method was better than most charge-assigning methods based on prediction accuracy, though it was not successful in yielding overall higher cross-validation correlation coefficient (q(2)) values than others. The CFF charge model worked the best in prediction accuracy when q(2) was used as the evaluation criterion. The results presented should help the selection of electrostatic potential models in CoMFA and CoMSIA studies.


Tailoring sub-3.3 Å ultramicropores in advanced carbon molecular sieve membranes for blue hydrogen production.
Carbon molecular sieve (CMS) membranes prepared by carbonization of polymers containing strongly size-sieving ultramicropores are attractive for high-temperature gas separations. However, polymers need to be carbonized at extremely high temperatures (900° to 1200°C) to achieve sub-3.3 Å ultramicroporous channels for H2/CO2 separation, which makes them brittle and impractical for industrial applications. Here, we demonstrate that polymers can be first doped with thermolabile cross-linkers before low-temperature carbonization to retain the polymer processability and achieve superior H2/CO2 separation properties. Specifically, polybenzimidazole (PBI) is cross-linked with pyrophosphoric acid (PPA) via H bonding and proton transfer before carbonization at ≤600°C. The synergistic PPA doping and subsequent carbonization of PBI increase H2 permeability from 27 to 140 Barrer and H2/CO2 selectivity from 15 to 58 at 150°C, superior to state-of-the-art polymeric materials and surpassing Robeson's upper bound. This study provides a facile and effective way to tailor subnanopore size and porosity in CMS membranes with desirable molecular sieving ability.


Lung respiration motion modeling: a sparse motion field presentation method using biplane x-ray images.
Respiration-introduced tumor location uncertainty is a challenge in the precise lung biopsy for lung lesions. Current statistical modeling approaches hardly capture the complex local respiratory motion information. In this study, we formulate a statistical respiratory motion model using biplane x-ray images to improve the accuracy of motion field estimation by efficiently preserving local motion details for specific patients. Given CT data sets of 18 healthy subjects at end-expiratory and end-inspiratory breathing phases, the respiratory motion field is constructed based on deformation vector fields which are extracted from these CT data sets, and a lung contour motion repository respiratory is generated dependent on displacements of boundary control points. By varying the sparse weight coefficients of the statistical sparse motion field presentation (SMFP) method, the newly-input motion field is approximately presented by a sparse linear combination of a subset of the motion repository. The SMFP method is employed twice in the coefficient optimization process. Finally, these non-zero coefficients are fine-tuned to maximize the similarity between the projection image of reconstructed volumetric images and the current x-ray image. We performed the proposed method for estimating respiratory motion field on ten subject datasets and compared the result with the PCA method. The maximum average target registration error of the PCA-based and the SMFP-based respiratory motion field estimation are 3.1(2.0) and 2.9(1.6) mm, respectively. The maximum average symmetric surface distance of two methods are 2.5(1.6) and 2.4(1.3) mm, respectively.


Oral Administration of a Hot Water Extract of the Softshell Turtle (Trionyx sinensis) Improves Exercise Performance.
Freshwater softshell turtle (Trionyx sinensis) extract has been used traditionally as a tonic soup, and to recover from physical fatigue. To support these claims, the forelimb grip strength of mice was measured after feeding a soft-shell turtle extract for 7 days. The T. sinensis extract significantly increased the grip strength to 1.25±0.07 N (P<0.01), which is 16.8% higher than the force on day 0. After exercising, the blood glucose levels in extract-fed mice were 202% higher and urea levels were 73% lower, which were both significantly different than the levels observed after control treatment. Lactate dehydrogenase was significantly higher by 314%, and glutathione peroxidase increased by 165%. In addition, the obesity markers, serum triglyceride and cholesterol, decreased to 62% and 49%, respectively, after mice were fed the extract. These data show that the T. sinensis extract provided more energy for forelimb exercise, prevented protein catabolism and muscle fatigue, and decreased the oxidative stress caused by an exhaustive workout.


Expression of human terminal deoxynucleotidyl transferase in Escherichia coli.
A cloned DNA fragment related to pT17 containing a partial cDNA sequence of human terminal deoxynucleotidyl transferase was used as a probe to screen for the full length cDNA sequence of the enzyme in a lambda gt11 library constructed from human lymphoblastoid KM-3 cDNA. A recombinant containing a 2068-base pair insert was isolated and recloned into the EcoRI site of the sequencing plasmic pUC-8 as two subclones, pT711 and pT106. DNA sequencing and hybridization studies showed that pT711 contains the pT17 sequence and an additional 172 upstream nucleotides. pT711 represents the coding sequence for the carboxyl half of the terminal transferase protein. pT106, containing a 965-base pair insert, hybridizes to the same mRNA as pT711 on Northern blots and contains an open reading frame that is in phase with the reading frame of the insert in pT711. Amino acid sequencing of the 58-kDa peptide of the calf thymus terminal transferase failed, indicating that the N terminus is blocked. N-Terminal sequencing of a 56-kDa form of the protein produced 24 amino acids corresponding to the translated human cDNA coding sequence starting at residue 398 of the insert in pT106 with 83% homology between bovine and human sequence. The initiation codon is assigned to an ATG sequence at nucleotide 329 of the insert in pT106. Comparison of the translated human terminal transferase sequence with peptides from the calf thymus enzyme showed that the homology between the human and bovine enzyme is better than 90% among 263 amino acids determined. The coding sequences in pT106 and pT711 were recloned into an expression plasmid pUC-19 downstream from the lac promoter and in phase with the coding sequence of the lac Z gene. Lysates of bacteria carrying the reconstructed coding sequence of human terminal transferase contain a fused protein of 60 kDa that reacts with rabbit antibody to terminal transferase on immunoblots and exhibits enzyme activity. Isolation of this fused protein from bacterial lysates with mouse monoclonal antibody to human terminal transferase produces the expected protein of 60 kDa.


Targeted Lipidomics Analysis of Oxylipids in Hazelnut Oil during Storage by Liquid Chromatography Coupled to Tandem Mass Spectrometry.
Hazelnut oil is a high-grade edible oil with high nutritional value and unique taste. However, it is prone to oxidative degradation during storage. Herein, we used liquid chromatography coupled to tandem mass spectrometry to carry out a lipidomics analysis of the storage process of hazelnut oil. A total of 41 triacylglycerols and 12 oxylipids were determined. The contents of all oxylipids increased significantly after storage (p < 0.05). The oxylipid accumulation of hazelnut oil during storage was clarified for the first time. Nine significantly different oxylipids were further screened out. It was considered that the 15th day of storage is the dividing point. In addition, the lipoxygenase-catalyzed oxidation may be the major contributor to lipid oxidation of hazelnut oil. This study provides a new insight and theoretical basis to explore the storage oxidation mechanism of hazelnut oil and take quality control measures.


Excess mannose limits the growth of phosphomannose isomerase PMI40 deletion strain of Saccharomyces cerevisiae.
Phosphomannose isomerase (PMI40) catalyzes the conversion between fructose 6-phosphate and mannose 6-phosphate and thus connects glycolysis, i.e. energy production and GDP-mannose biosynthesis or cell wall synthesis in Saccharomyces cerevisiae. After PMI40 deletion (pmi(-)) the cells were viable only if fed with extracellular mannose and glucose. In an attempt to force the GDP-mannose synthesis in the pmi(-) strain by increasing the extracellular mannose concentrations, the cells showed significantly reduced growth rates without any alterations in the intracellular GDP-mannose levels. To reveal the mechanisms resulting in reduced growth rates, we measured genome-wide gene expression levels, several metabolite concentrations, and selected in vitro enzyme activities in central metabolic pathways. The increasing of the initial mannose concentration led to an increase in the mannose 6-phosphate concentration, which inhibited the activity of the second enzyme in glycolysis, i.e. phosphoglucose isomerase converting glucose 6-phosphate to fructose 6-phosphate. As a result of this limitation, the flux through glycolysis was decreased as was the median expression of the genes involved in glycolysis. The expression levels of RAP1, a transcription factor involved in the regulation of the mRNA levels of several enzymes in glycolysis, as well as those of cell cycle regulators CDC28 and CLN3, decreased concomitantly with the growth rates and expression of many genes encoding for enzymes in glycolysis.