A LQR-Based Controller with Estimation of Road Bank for Improving Vehicle Lateral and Rollover Stability via Active Suspension.
In this article, a Linear Quadratic Regulator (LQR) lateral stability and rollover controller has been developed including as the main novelty taking into account the road bank angle and using exclusively active suspension for both lateral stability and rollover control. The main problem regarding the road bank is that it cannot be measured by means of on-board sensors. The solution proposed in this article is performing an estimation of this variable using a Kalman filter. In this way, it is possible to distinguish between the road disturbance component and the vehicle's roll angle. The controller's effectiveness has been tested by means of simulations carried out in TruckSim, using an experimentally-validated vehicle model. Lateral load transfer, roll angle, yaw rate and sideslip angle have been analyzed in order to quantify the improvements achieved on the behavior of the vehicle. For that purpose, these variables have been compared with the results obtained from both a vehicle that uses passive suspension and a vehicle using a fuzzy logic controller.


Recovery from Amblyopia in Adulthood: A Meta-Analysis.
The effectiveness of traditional amblyopia therapies is largely restricted to childhood. However, recovery in adulthood is possible following removal or vision-limiting disease of the fellow eye. Study of this phenomenon is currently limited to isolated case reports and a few case series, with reported incidence ranging from 19-77% 1-5 . We set out to accomplish two distinct goals: (1) define the incidence of clinically meaningful recovery and (2) elucidate the clinical features associated with greater amblyopic eye gains. A systematic review of 3 literature databases yielded 23 reports containing 109 cases of patients ≥18 years old with unilateral amblyopia and vision-limiting fellow eye pathology. Study 1 revealed 25/42 (59.5%) of adult patients gained ≥2 logMAR lines in the amblyopia eye after FE vision loss. The overall degree of improvement is clinically meaningful (median 2.6 logMAR lines). Study 2 showed that for cases with amblyopic eye visual acuity improvement, recovery occurs within 12 months of initial loss of fellow eye vision. Regression analysis revealed that younger age, worse baseline acuity in the amblyopic eye, and worse vision in the fellow eye independently conferred greater gains in amblyopic eye visual acuity. Recovery occurs across amblyopia types and fellow eye pathologies, although disease entities affecting fellow eye retinal ganglion cells demonstrate shorter latencies to recovery. Amblyopia recovery after fellow eye injury demonstrates that the adult brain harbors the neuroplastic capacity for clinically meaningful recovery, which could potentially be harnessed by novel approaches to treat adults with amblyopia.


Perioral skin biopsy to study skeletal muscle protein expression.
Clinical trials for muscular dystrophy molecular treatment require multiple sampling of skeletal muscle to monitor protein rescue. This practice is invasive and could raise ethical problems. A less invasive tool to obtain sequential muscle sampling is necessary. Using indirect immunofluorescence, we evaluated muscle protein expression in myofiber bundles included in 2-2.5-mm punch skin biopsies from the perioral region from 6 healthy subjects and 6 patients with genetically defined forms of muscular dystrophy. Large intradermal bundles of orbicularis oris muscle were constantly present in skin biopsies. They showed a typical muscular antigenic pattern in controls and the expected protein defect in muscular dystrophy patients. These results demonstrate the feasibility of muscular protein expression analysis using skin biopsy. We propose this minimally invasive technique to follow-up the response to genetic or conventional therapies in muscular dystrophies and to confirm the diagnosis in some special clinical conditions.


Distance dependence of electron transfer across peptides with different secondary structures: the role of Peptide energetics and electronic coupling.
The charge-transfer transition energies and the electronic-coupling matrix element, |H(DA)|, for electron transfer from aminopyridine (ap) to the 4-carbonyl-2,2'-bipyridine (cbpy) in cbpy-(gly)(n)-ap (gly = glycine, n = 0-6) molecules were calculated using the Zerner's INDO/S, together with the Cave and Newton methods. The oligopeptide linkages used were those of the idealized protein secondary structures, the alpha-helix, 3(10)-helix, beta-strand, and polyproline I- and II-helices. The charge-transfer transition energies are influenced by the magnitude and direction of the dipole generated by the peptide secondary structure. The electronic coupling |H(DA)| between (cbpy) and (ap) is also dependent on the nature of the secondary structure of the peptide. A plot of 2.ln|H(DA)| versus the charge-transfer distance (assumed to be the dipole moment change between the ground state and the charge-transfer states) showed that the polyproline II structure is a more efficient bridge for long-distance electron-transfer reactions (beta = 0.7 A(-1)) than the other secondary structures (beta approximately 1.3 A(-1)). Similar calculations on charged dipeptide derivatives, [CH(3)CONHCH(2)CONHCH(3)](+/)(-), showed that peptide-peptide interaction is more dependent on conformation in the cationic than in the anionic dipeptides. The alpha-helix and polyproline II-helix both have large peptide-peptide interactions (|H(DA)| > 800 cm(-1)) which arise from the angular dependence of their pi-orbitals. Such an interaction is much weaker than in the beta-strand peptides. These combined results were found to be consistent with electron-transfer rates experimentally observed across short peptide bridges in polyproline II (n = 1-3). These results can also account for directional electron transfer observed in an alpha-helical structure (different ET rates versus the direction of the molecular dipole).


Physical and functional analyses of the syrA and syrB genes involved in syringomycin production by Pseudomonas syringae pv. syringae.
The syrA and syrB genes involved in syringomycin production in Pseudomonas syringae pv. syringae B301D were identified from an EcoRI-pLAFR3 cosmid library and then physically and functionally analyzed in relation to plant pathogenicity. Homologous recombination of the genes required for syringomycin production from cosmids pGX183 (syrA) and pGX56 (syrB), respectively, introduced into nontoxigenic (Tox-) Tn5 mutants W4S2545 and W4S770 resulted in the concomitant restoration of toxin production and full virulence. The disease indices of the Tox+ strains obtained by recombination of the cloned, homologous DNA into the corresponding Tn5 mutant were essentially equivalent to that of strain B301D-R and significantly higher than those of W4S2545 and W4S770. A 12-kilobase (kb) EcoRI fragment from pGX183 was subcloned (i.e., pGX15) and found to contain the sequences necessary for syringomycin production. A map of pGX15 prepared by a combination of restriction endonuclease digestions and Tn5 mutagenesis showed that the syrA sequence was 2.3 to 2.8 kb. Marker exchange of syrA::Tn5 from pGX15 into B301D-R yielded nonpathogenic phenotypes, indicating that syrA is a regulatory gene since it is necessary for both syringomycin production and pathogenicity. The 4.9-kb EcoRI fragment from pGX56 was subcloned (i.e., pGX4) and shown to carry the syrB sequence which was 2.4 to 3.3 kb. Sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis of protein extracts from B301D-R associated five proteins, ranging from approximately 130,000 to approximately 470,000 in molecular weight, with syringomycin production. The syrA and syrB genes were required for the formation of proteins SR4 (approximately 350,000) and SR5 (approximately 130,000), which are believed to be components of the syringomycin synthetase complex.


Treatment of experimental autoimmune uveoretinitis with different natural compounds.
Uveitis is an important eye disease that potentially causes loss of sight. Although extensive studies have been conducted on uveitis, the exact pathogenesis remains to be determined. The effects of treatment with natural compounds on an experimental autoimmune uveoretinitis (EAU) rat model were examined in the present study. A total of 25 rats were divided into 5 groups: Alkaloids (n=5), saponins (n=5), flavonoids (n=5), phenols (n=5), and the normal saline group (n=5). The rats in each group were treated with an intraperitoneal injection of proper alkaloids (berberine hydrochloride), saponins (steroidal saponins), flavonoids (baicalein), or phenols (chlorogenic acid) or physiological saline, respectively. The rats' aqueous humour and crystalline lens was then observed under the slit lamp periodically, looking for signs of inflammation. After 2 weeks, the rats were sacrificed and the degree of pathological changes on their eyeballs under different treatment methods were determined using an optical microscope. The expression of the interleukin (IL)‑17 gene in the ocular tissues of the rats was assessed via RT‑PCR and western blot analysis. Apoptosis on the rats' retinal tissues was detected using flow cytometry. The results showed that rats injected with phenols (chlorogenic acid) had serious ocular vascular dilatation, iris hemorrhage and purulent exudation; those injected with alkaloids (berberine hydrochloride) and flavonoids (baicalein) had a more mild form of inflammation; and those administered saponins (steroidal saponins) had only mild inflammation signs. Following detection of IL‑17 mRNA and protein expression levels in the ocular tissues of rats of the five groups, it was found that their expression was lowest in the saponin‑treated group and the other differences in expression were all statistically significant (P<0.05). A comparison with other groups revealed that cell apoptosis in the eyes of rats in the saponin group was the most prominent, reflecting a beneficial decrease in the amount of inflammatory cells on the lesion. Based on these findings, natural compounds such as saponins (steroidal saponins), alkaloids (berberine hydrochloride), and flavonoids (baicalein), but not phenols (chlorogenic acid), can inhibit the clinical symptoms of EAU in rats to a certain extent and reduce cell apoptosis. The most promising results in the present study were obtained using steroidal saponins.


Pharmacokinetic analysis of gentamicin thrice and single daily dosage in pediatric cancer patients.
Fifty-two children suffering from different types of malignancies were included and evaluated for the pharmacokinetics of gentamicin thrice or single daily dosage protocols. All the study population received a total dose of 5 mg/kg daily. Thirty children received gentamicin thrice daily, and 22 were treated using the single daily protocol; all had fever and neutropenia when included. The individual pharmacokinetic parameters were calculated using a one-compartment model for two blood gentamicin samples. The mean (SD) t 1/2 (h), clearance (mL/min/BSA), Vss (L/kg), Cmax (micrograms/mL), and Cmin (micrograms/mL) were 3.05 (1.0) and 3.9 (0.6) h, 136 (61.3) and 99.9 (65.3) mL/min/BSA, 0.4035 (0.167) and 0.457 (0.17) L/kg, 5.2 (2.0) and 11.5 (4.2) micrograms/mL, 0.8 (0.6) and 0.18 (0.1) microgram/mL for thrice and single daily dosage schedules, respectively. The single gentamicin daily dose protocol had a significantly longer t 1/2, shorter clearance normalized to BSA, higher Cmax, and lower Cmin in comparison with the thrice daily schedule. We recommend the use of gentamicin at 5 mg/kg daily delivered as a single daily dose for pediatric cancer patients with fever and neutropenia, in spite of the measured pharmacokinetic differences, which in our opinion have no clinical significance.


Recombination between plasmids of incompatibility groups P-1 and P-2.
R plasmids of incompatibility group P-2 are readily transmissible between Pseudomonas strains, but not to Escherichia coli or other enterobacteria, whereas those of group P-1 have a broad host range. Pseudomonas aeruginosa donor strains carrying both a P-1 plasmid (RP1, RP4, or R751) and a P-2 plasmid (pMG1, pMG2, pMG5, or RPL11) were mated with E. coli K-12, and selection was imposed for resistance markers on the P-2 plasmids. Transconjugants were obtained at a low frequency, in which P-2 markers were expressed and were serially transmissible in E. coli together with P-1 markers. These plasmids had P-1 incompatibility properties, conferred susceptibility to phages active on P-1 carrying strains, and behaved on sucrose gradient centrifugation as unimolecular species of higher molecular weights than the P-1 parent. Recombinant plasmid formation was independent of a functional Rec gene in both donor and recipient and, with R751, had a preferred site leading to loss of trimethoprim resistance. Interaction between insertion sequences may be involved. Thus, plasmids of group P-2 can recombine with R factors of another group quite separate in compatibility properties, host range, and pilus type. Formation of such recombinants provides one pathway by which the genetic diversity of plasmids may have evolved.


Object fixation by the blowfly during tethered flight in a simulated three-dimensional environment.
The ability of flies to detect and fixate objects moving relative to their background was investigated in a flight simulator during translational tethered flight. The fly experienced optic flow that depended on its own actions and reactions in a similar way as in free-flight (closed-loop) conditions. Fixation of an object required turning responses towards it. The simulated distances between the fly, object and background were varied systematically by changing the velocities with which the object and the background pattern moved from the frontal to the back part of the fly's visual field. Fixation responses were only elicited when the object was simulated to be closer than the background. The fly's fixation performance was better with close than with more distant objects. Since, under many stimulus conditions, fixation responses were either elicited or entirely failed to be elicited, it is concluded that object fixation behaviour is gated in the visuo-motor pathway.


The Crosstalk of Adipose-Derived Stem Cells (ADSC), Oxidative Stress, and Inflammation in Protective and Adaptive Responses.
The potential use of stem cell-based therapies for the repair and regeneration of various tissues and organs is a major goal in repair medicine. Stem cells are classified by their potential to differentiate into functional cells. Compared with other sources, adipose-derived stem cells (ADSCs) have the advantage of being abundant and easy to obtain. ADSCs are considered to be tools for replacing, repairing, and regenerating dead or damaged cells. The capacity of ADSCs to maintain their properties depends on the balance of complex signals in their microenvironment. Their properties and the associated outcomes are in part regulated by reactive oxygen species, which mediate the oxidation-reduction state of cells as a secondary messenger. ADSC therapy has demonstrated beneficial effects, suggesting that secreted factors may provide protection. There is evidence that ADSCs secrete a number of cytokines, growth factors, and antioxidant factors into their microenvironment, thus regulating intracellular signaling pathways in neighboring cells. In this review, we introduce the roles of ADSCs in the protection of cells by modulating inflammation and immunity, and we develop their potential therapeutic properties.


alpha 1-adrenoceptor stimulation is able to reverse halothane-induced cardiac depression in isolated rat hearts.
Stimulation of myocardial alpha 1-adrenoceptors has been shown to exert positive inotropic effects through a cyclic AMP-independent mechanism. The purpose of this study was to examine if alpha 1-adrenoceptor stimulation is able to attenuate myocardial depression produced by exposure to halothane, and to test if alpha 1-adrenoceptor stimulation alters myocardial oxygen supply-demand balance in hearts exposed to halothane. The effects of phenylephrine were examined in 7 isolated perfused rat hearts. Variables measured were: heart rate, isovolumetric peak left ventricular pressure (LVP), LV dP/dt, coronary arterial flow, myocardial O2 delivery (DO2), myocardial O2 consumption (MVO2) and the ratio of DO2/MVO2. Each heart was exposed to phenylephrine cumulatively 0.1 microM, 0.3 microM, 1 microM and 3 microM under the administration of 1% halothane in the presence of propranolol 1 microM. Halothane 1% decreased the heart rate by 9 +/- 3%, LVP by 37 +/- 3%, and LV dP/dt by 35 +/- 2%. Phenylephrine restored these decreases to the baseline levels. Phenylephrine maintained or further enhanced the reductions in coronary flow and DO2 produced by halothane, resulting in a decrease in the DO2/ MVO2 ratio. alpha 1-adrenoceptor stimulation is capable of restoring direct cardiac depressant effects of halothane with a possible impairment of the oxygen supply-demand balance.


Perceptions toward HIV, HIV screening, and the use of antiretroviral medications: a survey of maternity-based health care providers in Zambia.
Mother-to-child transmission of HIV (MTCT) is a major contributor to Zambia's HIV burden. Based on our experience in Zambia, we felt that provider perceptions, knowledge base, and practice patterns toward HIV-positive mothers may pose as significant obstacles to preventing MTCT. Two hundred and twenty-five health care providers throughout Zambia were surveyed in 2002. Providers reported widespread stigma associated with HIV. Physicians (OR = 1.9), providers with research affiliations (OR = 2.3), and those located in Lusaka (OR = 9.0) were more likely to offer HIV testing. Only 30% routinely prescribed antiretroviral treatment (ART) to reduce MTCT. Practitioners from district facilities, those from Lusaka, and those employed at research facilities were more likely to prescribe ART routinely (OR = 2.8, 10.1 and 3.4 respectively). Among those never prescribing ART, most cited a lack of availability (83%). Our results highlight the need for further provider education, critical appraisal of the current system for HIV testing, and widespread distribution of ART.


[A man with an abnormal dimple in his upper leg].
A 29 year-old man presented with muscle weakness and an abnormal dimple near his hamstrings after a complicated anterior cruciate ligament reconstruction of his right knee. MRI demonstrated atrophy of the semitendinosus muscle. He received physiotherapy after which he functionally recovered.


Cloning and analysis of human gastric mucin cDNA reveals two types of conserved cysteine-rich domains.
Human gastric mucin was isolated by successive CsCl-gradient ultracentrifugation in the presence of guanidinium hydrochloride to prevent degradation of the polypeptide moieties of the molecules. The amino acid sequence of a tryptic fragment of this molecule was identical to that of a tryptic fragment of tracheobronchial mucin. An oligonucleotide based on this sequence hybridized specifically to human stomach mRNA and was subsequently used to screen a human stomach lambda ZAPII cDNA library. The largest of 10 positive clones encoded 850 amino acid residues, including the tryptic fragment, with high amounts of threonine, serine and proline residues. Interestingly, cysteine accounted for almost 8% of the amino acid residues. The 3' part of the sequence was very similar but not identical to the 3' region of human tracheobronchial cDNA. No tandem repeated sequences were present and the deduced polypeptide sequence contained two potential N-linked glycosylation sites. Four cysteine-rich clusters were detected, one of which was apparently homologous to the D-domains present in other mucins and in von Willebrand factor. The arrangement of the cysteines in three other cysteine-rich clusters was conserved in the human gastric mucin cDNA in a similar fashion as in two domains in the MUC2 gene product. The cysteine-rich domains were separated by short stretches of non-repetitive amino acid residues with a very high content of threonine and serine residues. These data suggest that the encoded polypeptide of this clone may be involved in disulphide-bond-mediated oligomerization of the mucin, and provide new insights into the molecular organization of mammalian apomucins.


Problem-based versus conventional curricula: influence on knowledge and attitudes of medical students towards health research.
Medical education curricula in developing countries should emphasize training in health research. This study compares the knowledge and attitudes towards health research between undergraduate medical students undertaking Problem Based Learning (PBL) versus conventional Lecture Based Learning (LBL). Two groups comprising 66 (LBL) and 84 (PBL) 4(th) and 5(th) year students from the medical college of Aga Khan University were administered a structured and validated questionnaire. Knowledge and attitudes of the two groups were recorded on a scale (graduated in percentages) and compared for statistical difference. PBL students scored 54.0% while LBL students scored 55.5% on the knowledge scale [p-value; 0.63]. On the attitudes scale, PBL students scored 75.5% against a 66.7% score of LBL students [p-value; 0.021]. A higher proportion of PBL students (89%) had participated in research activities compared to LBL students (74%) and thus felt more confident in conducting research and writing a scientific paper. The PBL students showed slightly healthier attitudes towards health research compared to LBL students. Both groups demonstrated a similar level of knowledge about health research. The positive impact of the PBL curriculum on attitudes of medical students towards health research may help in improving research output from developing countries in future.


Vincristine-induced neuropathy as the initial presentation of charcot-marie-tooth disease in acute lymphoblastic leukemia: a Pediatric Oncology Group study.
After profound peripheral neurotoxicity during induction chemotherapy for acute lymphoblastic leukemia (ALL) in the index patient with Charcot-Marie-Tooth hereditary neuropathy (CMT), study coordinators of the Pediatric Oncology Group (POG) front-line ALL protocols reviewed patient registrations to identify any other patients with possible CMT. The goal was to provide preliminary information about patients with undiagnosed CMT who develop ALL. Five children with ALL who were enrolled in POG B-precursor or T-cell ALL protocols from 1994 to 1999 subsequently were determined to have CMT hereditary neuropathy. Their clinical presentations and treatment records were reviewed in detail. Records of all patients entered on POG 9201 (lesser-risk ALL) were reviewed to identify all cases of significant vincristine toxicity noted in the first 6 months of treatment. The five identified patients all had substantial peripheral neurotoxicity that required alteration in treatment and/or orthopedic/physical therapy evaluation and follow-up. The POG 9201 review identified 25 of 686 patients (3.6%) with significant peripheral neuropathy. Three of 25 were diagnosed with CMT; the others have had no testing reported. A family history of CMT or other peripheral neuropathy should be sought at the time of diagnosis of ALL. Testing for CMT should be considered in any child with substantial vincristine-induced peripheral neurotoxicity. Treatment of such patients must be individualized. Testing of all patients with significant peripheral neuropathy would be necessary to determine the percentage of such neuropathy explained by underlying CMT.


Mast cell survival and mediator secretion in response to hypoxia.
Tissue hypoxia is a consequence of decreased oxygen levels in different inflammatory conditions, many associated with mast cell activation. However, the effect of hypoxia on mast cell functions is not well established. Here, we have investigated the effect of hypoxia per se on human mast cell survival, mediator secretion, and reactivity. Human cord blood derived mast cells were subjected to three different culturing conditions: culture and stimulation in normoxia (21% O(2)); culture and stimulation in hypoxia (1% O(2)); or 24 hour culture in hypoxia followed by stimulation in normoxia. Hypoxia, per se, did not induce mast cell degranulation, but we observed an increased secretion of IL-6, where autocrine produced IL-6 promoted mast cell survival. Hypoxia did not have any effect on A23187 induced degranulation or secretion of cytokines. In contrast, cytokine secretion after LPS or CD30 treatment was attenuated, but not inhibited, in hypoxia compared to normoxia. Our data suggests that mast cell survival, degranulation and cytokine release are sustained under hypoxia. This may be of importance for host defence where mast cells in a hypoxic tissue can react to intruders, but also in chronic inflammations where mast cell reactivity is not inhibited by the inflammatory associated hypoxia.


Furosemide Stress Test and Biomarkers for the Prediction of AKI Severity.
Clinicians have access to limited tools that predict which patients with early AKI will progress to more severe stages. In early AKI, urine output after a furosemide stress test (FST), which involves intravenous administration of furosemide (1.0 or 1.5 mg/kg), can predict the development of stage 3 AKI. We measured several AKI biomarkers in our previously published cohort of 77 patients with early AKI who received an FST and evaluated the ability of FST urine output and biomarkers to predict the development of stage 3 AKI (n=25 [32.5%]), receipt of RRT (n=11 [14.2%]), or inpatient mortality (n=16 [20.7%]). With an area under the curve (AUC)±SEM of 0.87±0.09 (P<0.0001), 2-hour urine output after FST was significantly better than each urinary biomarker tested in predicting progression to stage 3 (P<0.05). FST urine output was the only biomarker to significantly predict RRT (0.86±0.08; P=0.001). Regardless of the end point, combining FST urine output with individual biomarkers using logistic regression did not significantly improve risk stratification (ΔAUC, P>0.10 for all). When FST urine output was assessed in patients with increased biomarker levels, the AUC for progression to stage 3 improved to 0.90±0.06 and the AUC for receipt of RRT improved to 0.91±0.08. Overall, in the setting of early AKI, FST urine output outperformed biochemical biomarkers for prediction of progressive AKI, need for RRT, and inpatient mortality. Using a FST in patients with increased biomarker levels improves risk stratification, although further research is needed.


Association between physical activity, adiposity, and lipid abnormalities in children with familial hyperlipidemia.
The benefits of physical activity in children have been studied extensively; however, its role in children with familial hyperlipidemia (FH) is unknown. To determine associations between physical activity, adiposity, and lipid profiles in children with FH. A physical activity questionnaire was completed by 147 children with FH. Correlations between activity levels, body mass index (BMI), and fasting lipid profiles were determined. The mean age of patients was 12.5+/-3.2 years with a mean total cholesterol of 6.17 mmol/l (238 mg/dl), low-density lipoprotein-cholesterol of 4.43 (171), high-density lipoprotein-cholesterol of 1.08 (42), and triglyceride levels of 1.51 (134). Patients had greater weight for height indices than normal, with a mean BMI z score of +0.90+/-1.30 SD (P<0.001 versus normal), and with 21% of the participants being more than 2 SD above normal. Higher BMI z scores significantly correlated with higher triglyceride levels (r=0.33; P<0.0001) and greater time spent in sedentary pursuits (r=0.24; P=0.004), in particular watching television (r=0.26; P=0.003). The increased time that other family members spent in physical activity significantly correlated with a lower BMI z score (r=-0.21; P=0.01) of the patient and greater time spent in physical activity (r=0.24; P=0.003). There was no association between patients' physical activity levels and lipid profile or BMI. Similar to the general population, children with FH are also at risk of becoming overweight. Increased adiposity significantly correlated with the greater sedentary activities of the patient, lower physical activities of the family, and higher triglyceride levels. Physical activity levels of the patient correlated with family activity levels.


Evaluation of Michael-type acceptor reactivity of 5-benzylidenebarbiturates, 5-benzylidenerhodanines, and related heterocycles using NMR.
Despite existing experimental and computational tools to assess the risk, the non-specific chemical modification of protein thiol groups remains a significant source of false-positive hits, particularly in academic drug discovery. Herein, we describe the application of a simple NMR method in a systematic study on the reactivity of 5-benzylidenebarbiturates, 5-benzylidenerhodanines, and their related oxo-heterocycles, which have been associated with numerous biological activities and have recently gained a reputation as unselective promiscuous binders. Using this method, we confirmed the reactivity of 5-benzylidenebarbiturates, which are known to easily form Michael adducts with nucleophiles. In contrast, 5-benzylidene five-membered oxo-heterocycles revealed almost insignificant reactivity. We can conclude that the distinct binding profile of the most controversial compounds, 5-benzylidenerhodanines, is not necessarily related to their unspecific Michael acceptor reactivity.


Development of a Cardiac Anesthesia Tool (CAT) to Predict Intensive Care Unit (ICU) Admission for Pediatric Cardiac Patients Undergoing Non-cardiac Surgery: A Retrospective Cohort Study.
of the work: Pediatric cardiac patients often undergo non-cardiac surgical procedures and many of these patients would require intensive care unit admission, but can we predict the need for ICU admission in pediatric cardiac patients undergoing non-cardiac procedures. Numerous preoperative and intraoperative variables were strongly associated with ICU admission. Given the variations in the underlying cardiac physiology and the diversity of noncardiac surgical procedures along with the scarce predictive clinical tools, we aimed to develop a simple and practical tool to predict the need for ICU admission in pediatric cardiac patients undergoing non-cardiac procedures. This is a retrospective study, where all files of pediatric cardiac patients who underwent noncardiac surgical procedures from January 1, 2015, to December 31, 2019, were reviewed. We retrieved details of the preoperative and intraoperative variables including age, weight, comorbid conditions, and underlying cardiac physiology. The primary outcome was the need for ICU admission. We performed multiple logistic regression analyses and analyses of the area under receiver operating characteristics (ROC) curves to develop a predictive tool. In total, 519 patients were included. The mean age and weight were 4.6 ± 3.4 year and 16 ± 13 Kg respectively. A small proportion (n = 90, 17%) required ICU admission. Statistically, there was strong association between each of American society of anesthesiologist's physical status (ASA-PS) class III and IV, difficult intubation, operative time more than 2 hours, requirement of transfusion and the failure of a deliberately planned extubation and ICU admission. Additional analysis was done to develop a Cardiac Anesthesia Tool (CAT) based on the weight of each variable derived from the regression coefficient. The CAT list is composed of the ASA-PS, operative time, and requirement of transfusion, difficult intubation and the failure of deliberately planned extubation. The minimum score is zero and the maximum is eight. The probability of ICU admission is proportional to the score. CAT is a practical and simple clinical tool to predict the need for ICU admission based on simple additive score. We propose using this tool for pediatric cardiac patients undergoing non-cardiac procedure.


Role of glial filaments in cells and tumors of glial origin: a review.
In the adult human brain, normal astrocytes constitute nearly 40% of the total central nervous system (CNS) cell population and may assume a star-shaped configuration resembling epithelial cells insofar as the astrocytes remain intimately associated, through their cytoplasmic extensions, with the basement membrane of the capillary endothelial cells and the basal lamina of the glial limitans externa. Although their exact function remains unknown, in the past, astrocytes were thought to subserve an important supportive role for neurons, providing a favorable ionic environment, modulating extracellular levels of neurotransmitters, and serving as spacers that organize neurons. In immunohistochemical preparations, normal, reactive, and neoplastic astrocytes may be positively identified and distinguished from other CNS cell types by the expression of the astrocyte-specific intermediate filament glial fibrillary acidic protein (GFAP). Glial fibrillary acidic protein is a 50-kD intracytoplasmic filamentous protein that constitutes a portion of, and is specific for, the cytoskeleton of the astrocyte. This protein has proved to be the most specific marker for cells of astrocytic origin under normal and pathological conditions. Interestingly, with increasing astrocytic malignancy, there is progressive loss of GFAP production. As the human gene for GFAP has now been cloned and sequenced, this review begins with a summary of the molecular biology of GFAP including the proven utility of the GFAP promoter in targeting genes of interest to the CNS in transgenic animals. Based on the data provided the authors argue cogently for an expanded role of GFAP in complex cellular events such as cytoskeletal reorganization, maintenance of myelination, cell adhesion, and signaling pathways. As such, GFAP may not represent a mere mechanical integrator of cellular space, as has been previously thought. Rather, GFAP may provide docking sites for important kinases that recognize key cellular substrates that enable GFAP to form a dynamic continuum with microfilaments, integrin receptors, and the extracellular matrix.


Tandemly Integrated HPV16 Can Form a Brd4-Dependent Super-Enhancer-Like Element That Drives Transcription of Viral Oncogenes.
In cancer cells associated with human papillomavirus (HPV) infections, the viral genome is very often found integrated into the cellular genome. The viral oncogenes E6 and E7 are transcribed from the viral promoter, and integration events that alter transcriptional regulation of this promoter contribute to carcinogenic progression. In this study, we detected highly enriched binding of the super-enhancer markers Brd4, MED1, and H3K27ac, visible as a prominent nuclear focus by immunofluorescence, at the tandemly integrated copies of HPV16 in cells of the cervical neoplasia cell line W12 subclone 20861. Tumor cells are often addicted to super-enhancer-driven oncogenes and are particularly sensitive to disruption of transcription factor binding to the enhancers. Treatment of 20861 cells with bromodomain inhibitors displaced Brd4 from the HPV integration site, greatly decreased E6/E7 transcription, and inhibited cellular proliferation. Thus, Brd4 activates viral transcription at this integration site, and strong selection for E6/E7 expression can drive the formation of a super-enhancer-like element to promote oncogenesis. Oncogenic human papillomaviruses play an essential role in the development of cervical cancer, and growth of these cancer cells requires continued expression of the viral E6 and E7 oncogenes. Integration of the virus into the host genome often results in deregulation of E6 and E7 expression, which provides a selective growth advantage and increases genetic instability of infected cells. We show here that tandemly integrated copies of the viral genome can form a super-enhancer-like element that drives E6/E7 transcription. Targeted disruption of factors binding to this element decreases viral transcription and causes cell death. Thus, cancer cells that harbor integrated HPV could be targeted by therapeutics that disrupt super-enhancer function.


SIRT1 is associated with a decrease in acute insulin secretion and a sex specific increase in risk for type 2 diabetes in Pima Indians.
Genetic variation in SIRT1 affects obesity-related phenotypes in several populations. The purpose of this study was to determine whether variation in SIRT1 affects susceptibility to obesity or type 2 diabetes in Pima Indians, a population with very high prevalence and incidence rates of these diseases. Genotypic data from single nucleotide polymorphisms (SNPs) identified by sequencing regions of SIRT1 combined with SNPs in/near SIRT1 from a prior genome-wide association study determined that 4 tag SNPs (rs7895833, rs10509291, rs7896005, and rs4746720) could capture information across this gene and its adjacent 5' region. The tag SNPs were genotyped in a population-based sample of 3501 Pima Indians (44% had diabetes, 58% female) for association with type 2 diabetes and BMI. Metabolic trait data and adipose biopsies were available on a subset of these subjects. Two tag SNPs, rs10509291 and rs7896005, were nominally associated with type 2 diabetes (P=0.01, OR=1.25 95%CI 1.05-1.48, and P=0.02, OR=1.17 95%CI 1.02-1.34, respectively; additive P values adjusted for age, sex, birth year, and family membership), but not BMI (adjusted P values 0.52 and 0.45, respectively). Among metabolically characterized subjects with normal glucose tolerance (N=243), those carrying the diabetes risk allele (T) for rs10509291 and (G) for rs7896005 had a reduced acute insulin response (AIR) to an intravenous glucose bolus (adjusted P=0.045 and 0.035, respectively). SIRT1 expression in adipose biopsies was negatively correlated with BMI (adjusted P=0.00001). We conclude that variation in SIRT1 is nominally associated with reduced AIR and increased risk for type 2 diabetes. SIRT1 expression in adipose is correlated with BMI, but it remains unknown whether this is a cause or consequence of obesity.


[Autonomic homeostasis of children born to parents irradiated in the result of the Chernobyl catastrophe with isolated abnormal chords of the left ventricle].
156 children of the main group (were born to parents irradiated in the result of Chernobyl disaster), in which according to Doppler echocardiography isolated abnormal chords of the left ventricle (AHLV) identified, 44 children of control group and 50 children of referent group were involved in the study Analysis of indicators of operational regulation of heart rate in children of primary group indicate the existence of autonomic dysregulation, which has the maximum expression in children with a threshold number of isolated abnormal chords. Evidence of autonomic dysregulation in children born to parents irradiated in the result of the Chernobyl disaster with isolated AHLV requires primary and secondary prevention to reduce the frequency of unfavourable clinical course.


WLP3 Encodes the Ribosomal Protein L18 and Regulates Chloroplast Development in Rice.
Plastid ribosomal proteins play a crucial role in the growth and development of plants, mainly in the gene expression and translation of key genes in chloroplasts. While some information is known about the regulatory processes of plastid ribosomal proteins in various plant species, there is limited knowledge about the underlying mechanisms in rice. In this study, ethyl methanesulfonate (EMS) mutagenesis was used to generate a new mutant called wlp3 (white leaf and panicle3), characterized by white or albino leaves and panicles, which exhibited this phenotype from the second leaf stage until tillering. Furthermore, after a certain period, the newly emerging leaves developed the same phenotype as the rice variety ZH11, while the albino leaves of wlp3 showed an incomplete chloroplast structure and significantly low chlorophyll content. A transition mutation (T to C) at position 380 was identified in the coding region of the LOC_Os03g61260 gene, resulting in the substitution of isoleucine by threonine during translation. WLP3 encodes the ribosomal L18 subunit, which is localized in the chloroplast. Complementation experiments confirmed that LOC_Os03g61260 was responsible for the albino phenotype in rice. WLP3 has high expression in the coleoptile, leaves at the three-leaf stage, and panicles at the heading stage. Compared to the wild-type (WT), wlp3 exhibited reduced chlorophyll synthesis and significantly decreased expression levels of genes associated with plastid development. Yeast two-hybrid (Y2H) analysis revealed that WLP3 interacts with other ribosomal subunits, to influence chloroplast development. These results contribute to a better understanding of the underlying molecular mechanisms of chloroplast development and plastid gene translation.


A water extract of Mucuna pruriens provides long-term amelioration of parkinsonism with reduced risk for dyskinesias.
Dopaminergic anti-parkinsonian medications, such as levodopa (LD) cause drug-induced dyskinesias (DID) in majority of patients with Parkinson's disease (PD). Mucuna pruriens, a legume extensively used in Ayurveda to treat PD, is reputed to provide anti-parkinsonian benefits without inducing DID. We compared the behavioral effects of chronic parenteral administration of a water extract of M. pruriens seed powder (MPE) alone without any additives, MPE combined with the peripheral dopa-decarboxylase inhibitor (DDCI) benserazide (MPE+BZ), LD+BZ and LD alone without BZ in the hemiparkinsonian rat model of PD. A battery of behavioral tests assessed by blinded investigators served as outcome measures in these randomized trials. In experiment 1, animals that received LD+BZ or MPE+BZ at high (6mg/kg) and medium (4mg/kg) equivalent doses demonstrated significant alleviation of parkinsonism, but, developed severe dose-dependent DID. LD+BZ at low doses (2mg/kg) did not provide significant alleviation of parkinsonism. In contrast, MPE+BZ at an equivalent low dose significantly ameliorated parkinsonism. In experiment 2, MPE without any additives (12mg/kg and 20mg/kg LD equivalent dose) alleviated parkinsonism with significantly less DID compared to LD+BZ or MPE+BZ. In experiment 3, MPE without additives administered chronically provided long-term anti-parkinsonian benefits without causing DID. In experiment 4, MPE alone provided significantly more behavioral benefit when compared to the equivalent dose of synthetic LD alone without BZ. In experiment 5, MPE alone reduced the severity of DID in animals initially primed with LD+BZ. These findings suggest that M. pruriens contains water-soluble ingredients that either have an intrinsic DDCI-like activity or mitigate the need for an add-on DDCI to ameliorate parkinsonism. These unique long-term anti-parkinsonian effects of a parenterally administered water extract of M. pruriens seed powder may provide a platform for future drug discoveries and novel treatment strategies in PD.


Dengue and Typhoid Co-infection- Study from a Government Hospital in North Delhi.
Concurrent infection with two agents can result in an illness having overlapping symptoms creating a diagnostic dilemma for the treating physician. The symptoms of dengue may mimic other diseases such as leptospirosis, influenza A, Salmonella Typhi, Japanese encephalitis, chikungunya and malaria. There is paucity of data regarding dengue and typhoid co- infection both in the developed and developing countries. This study attempts to find the current co- infection rates in North Delhi. This retrospective study was done between August and November 2013. Medical records of 659 patients exhibiting febrile illness who visited Kasturba Hospital were studied. Dengue specific IgM antibodies were detected by Dengue IgM antibody capture ELISA test. Serodiagnosis of Salmonella infection was conducted by Widal test in the hospital. Of the 659 febrile sera samples tested here, 141 (21.39%) tested positive for dengue. Of these 91 were females and 50 males. Of the dengue cases, eleven were co-infected with enteric fever (11/141= 7.8%). Maximum number of dengue positive cases seen in age group 0-10 y. Case Fatality Rate (CFR) was zero. Age groups of patients co-infected with dengue and typhoid were as follows: 0-10 y: 5, 11-20 y: 3, 21-30 y: 2, >60 y: 1. Co-infection should always be kept in mind while dealing with cases of dengue or enteric fever with or without atypical features. In order to reduce the burden of disease, along with improvement of sanitation and personal hygiene, emphasis should be given on vaccination against typhoid.


Inhibition of aquaporin-1 water permeability by tetraethylammonium: involvement of the loop E pore region.
Previously, the only known blockers of water permeability through aquaporin-1 (AQP1) water channels were mercurial reagents such as HgCl(2). For AQP1, inhibition by mercury has been attributed to the formation of a mercaptide bond with cysteine residue 189 found in the putative pore-forming region loop E. Here we show that the nonmercurial compound, tetraethylammonium (TEA) chloride, reduces the water permeability of human AQP1 channels expressed in Xenopus oocytes. After preincubation of the oocytes for 15 min with 100 microM TEA, AQP1 water permeability was reduced by 20 to 40%, a degree of partial block similar to that obtained with 15 min of incubation in 100 microM HgCl(2). The reduction of water permeability was dose-dependent for tested concentrations up to 10 mM TEA. TEA blocks the Shaker potassium channel by interacting with a tyrosine residue in the outer pore region. We tested whether an analogous tyrosine residue in loop E of AQP1 could be involved in the binding of TEA. Using polymerase chain reaction, tyrosine 186 in AQP1, selected for its proximity to the mercury-binding site, was mutated to phenylalanine (Y186F), alanine (Y186A), or asparagine (Y186N). Oocyte expression of the mutant AQP1 channels showed that the water permeability of Y186F was equivalent to that of wild-type AQP1; the other mutant channels did not conduct water. However, in contrast to wild-type AQP1, the water permeability of Y186F was not reduced with 100 microM TEA. These results suggest that TEA reduces AQP1 water permeability by interacting with loop E.


Oncolytic adenovirus ICOVIR-7 in patients with advanced and refractory solid tumors.
Twenty-one patients with cancer were treated with a single round of oncolytic adenovirus ICOVIR-7. ICOVIR-7 features an RGD-4C modification of the fiber HI-loop of serotype 5 adenovirus for enhanced entry into tumor cells. Tumor selectivity is mediated by an insulator, a modified E2F promoter, and a Rb-binding site deletion of E1A, whereas replication is optimized with E2F binding hairpins and a Kozak sequence. ICOVIR-7 doses ranged from 2 x 10(10) to 1 x 10(12) viral particles. All patients had advanced and metastatic solid tumors refractory to standard therapies. ICOVIR-7 treatment was well tolerated with mild to moderate fever, fatigue, elevated liver transaminases, chills, and hyponatremia. One patient had grade 3 anemia but no other serious side effects were seen. At baseline, 9 of 21 of patients had neutralizing antibody titers against the ICOVIR-7 capsid. Treatment resulted in neutralizing antibody titer induction within 4 weeks in 16 of 18 patients. No elevations of serum proinflammatory cytokine levels were detected. Viral genomes were detected in the circulation in 18 of 21 of patients after injection and 7 of 15 of the samples were positive 2 to 4 weeks later suggesting viral replication. Overall, objective evidence of antitumor activity was seen in 9 of 17 evaluable patients. In radiological analyses, 5 of 12 evaluable patients had stabilization or reduction in tumor size. These consisted of one partial response, two minor responses and two cases of stable disease, all occurring in patients who had progressive disease before treatment. In summary, ICOVIR-7 treatment is apparently safe, resulting in anticancer activity, and is therefore promising for further clinical testing.


Medicare Advantage: Growth Amidst Mounting Scrutiny.
The Medicare Advantage Program, home to nearly half of the eligible Medicare population, has recently come under increased scrutiny. The Government Accountability Office called on the Centers for Medicare & Medicaid Services to monitor "disenrollment of MA beneficiaries in the last year of life, validate MA-provided encounter data, and strengthen audits used to identify and recover improper payments to MA plans." The House Subcommittee on Oversight and Investigations of the Committee on Energy & Commerce, dedicated a hearing to "Protecting America's Seniors: Oversight of Private Sector Medicare Advantage Plans." In addition, a recently conducted audit of the Office of the Inspector General of the Department of Health and Human Services raised concerns over "denials of prior authorization requests" and "beneficiary access to medically necessary care." In this article we consider the backdrop for the growing scrutiny of the MA program and the implications thereof to its future trajectory.


Significance of exercise-induced ST-segment elevation in patients without myocardial infarction.
Sixteen patients with exercise-induced ST-segment elevation and without a history of myocardial infarction or left ventricular aneurysm were studied. Fourteen complained of angina at rest, which was associated with ST-segment elevation in the same leads where it was recorded during exercise, and two patients had only exertional angina. Exercise-induced ST-segment elevation was generally reproducible in subsequent exercise tests performed in different hours of the day, but exercise tests repeated a mean of 15 months later did not induce this electrocardiographic abnormality. All patients had a marked susceptibility to coronary spasm, as shown by the response to the ergonovine test (12 positive tests in 12 patients) and by the occurrence of spontaneous spasm during coronary arteriography in two patients. In addition, coronary arteriography, performed in seven patients at the time of exercise-induced ST-segment elevation, revealed spasm of a major coronary vessel in all. In two patients we documented that exercise-induced ST-segment elevation was accompanied by a decreased coronary blood flow and increased coronary vascular resistance. We conclude that exercise-induced ST-segment elevation in patients without a history of myocardial infarction or left ventricular aneurysm is caused by coronary spasm of a major coronary vessel.


Sleep, psychological distress, and stress arousal in women with fibromyalgia.
The purpose of this investigation was to compare self-reported sleep quality and psychological distress, as well as somnographic sleep and physiological stress arousal, in women recruited from the community with self-reported medically diagnosed fibromyalgia (FM) to women without somatic symptoms. Eleven midlife women with FM, when compared to 11 asymptomatic women, reported poorer sleep quality and higher SCL-90 psychological distress scores. Women with FM also had more early night transitional sleep (stage 1) (p < 0.01), more sleep stage changes (p < 0.03) and a higher sleep fragmentation index (p < 0.03), but did not differ in alpha-EEG-NREM activity (a marker believed to accompany FM). No physiological stress arousal differences were evident. Less stable sleep in the early night supports a postulate that nighttime hormone (e.g., growth hormone) disturbance is an etiologic factor but, contrary to several literature assertions, alpha-EEG-NREM activity sleep does not appear to be a specific marker of FM. Further study of mechanisms is needed to guide treatment options.


Health Promoting Behaviors in a Population-based Sample of Middle-aged Women and its Relevant Factors in Yazd, Iran.
This study is carried out with the aim of describing the pattern of health-promoting behaviors (HPBs) among middle-aged women and its relevant factors in Iran as a Muslim developing country. In a cross-sectional study, 483 middle-aged women were selected using cluster random sampling from Yazd city in Central Iran. An adjusted Persian version of Health Promoting Lifestyle Profile II was used for measuring HPBs. The demographic data were collected too. The average score of middle-aged women HPBs was 2.73 ± 0.36 (2.69 ± 2.76, %95). Spiritual growth and physical activity had the highest and the lowest score in HPBs' dimensions, respectively. HPBs had a positive significant correlation with age (p=0.02) and education level (p=0.001) and a negative significant correlation with number of children (p=0.005). HPBs had a higher level in retired women than employees and housewives. In spite of a desirable level of HPBs in spiritual growth and interpersonal relationship dimensions among this group, their level of HPBs in physical activity and health responsibility dimensions was undesirable. It means that the feature of health among Iranian middle-aged women in a Muslim developing country is different from the industrialized developed and also the non-Muslim countries; hence, they need a different intervention programs.


Rationale and methodological options for assessing infectious disease and related measures in social science surveys.
Infectious disease is an important, but often overlooked, component of population health in high-income nations. Common, everyday infections exact significant costs, including school and work absenteeism, reduced productivity, and substantial health care expenditures. Infectious disease also shapes trajectories of biological risk and health and may be causally linked to chronic disease risk later in life. The size, diversity, and representativeness of samples typically employed in survey-based studies of health present exceptional opportunities for advancing scientific knowledge on the social and economic determinants of infectious disease in childhood and adulthood and to investigate the long-term consequences of infectious disease for well-being and attainment across multiple domains. A wide range of interview-based, anthropometric, and biomarker measurement options are currently available for assessing infectious exposures, inflammation, and immune function in nonclinical settings. These methods afford opportunities for innovative, transdisciplinary research on the causes and consequences of infectious disease across the life course that can address questions of interest to social, life, and biomedical scientists.


Impact of treatment modality on clinical outcome in metastatic colorectal cancer patients stratified by metastatic sites.
To investigate the impact of location of metastases, and therapeutic modality on clinical outcomes in patients with metastatic colorectal cancer (CRC). Data for metastatic CRC patients were sourced from the Surveillance, Epidemiology, and End Results (SEER) database (SEER ID: 15309-Nov2017). Patients were classified as follows: Group 1 patients had only liver metastasis; Group 2 patients had liver and lung metastasis; Group 3 patients had more than two metastasis sites. Patients were treated with surgery alone, radiation alone, or surgery plus radiation. The main study outcomes were (1) cancer-specific mortality and (2) survival benefit associated with treatment modality. A total number of 15,510 patients were included in this study. In Groups 1 and 2, patients treated with surgery plus radiation had a higher cumulative survival compared to other treatment groups (p-value <.001). Group 3 patients showed no significant difference in cumulative survival between the different treatment modalities (p-value = .218). Group 1 patients who received surgery plus radiation had a significantly lower risk of mortality compared to the other treatment groups (p-value <.001), and Group 2 patients who either received radiation treatment alone or surgery plus radiation had a significantly lower risk for mortality than patients who received other treatment modalities (p-value <.001). Multivariate analysis adjusting for known prognostic factors such as tumor sidedness and race did not alter the observed risk conferred by metastasis sites and treatment modalities. Stratification by metastases sites, and by treatment modality can help multidisciplinary teams to reach a treatment consensus for metastatic CRC.


Unimolecular Decomposition Reactions of Picric Acid and Its Methylated Derivatives─A DFT Study.
To handle energetic materials safely, it is important to have knowledge about their sensitivity. Density functional theory (DFT) has proven a valuable tool in the study of energetic materials, and in the current work, DFT is employed to study the thermal unimolecular decomposition of 2,4,6-trinitrophenol (picric acid, PA), 3-methyl-2,4,6-trinitrophenol (methyl picric acid, mPA), and 3,5-dimethyl-2,4,6-trinitrophenol (dimethyl picric acid, dmPA). These compounds have similar molecular structures, but according to the literature, mPA is far less sensitive to impact than the other two compounds. Three pathways believed important for the initiation reactions are investigated at 0 and 298.15 K. We compare the computed energetics of the reaction pathways with the objective of rationalizing the unexpected sensitivity behavior. Our results reveal a few if any significant differences in the energetics of the three molecules, and thus do not reflect the sensitivity deviations observed in experiments. These findings point toward the potential importance of crystal structure, crystal morphology, bimolecular reactions, or combinations thereof on the impact sensitivity of nitroaromatics.


Diet and gallstones in Italy: the cross-sectional MICOL results.
Fifteen thousand nine hundred ten men and 13,674 women (age, 30-69 years) were enrolled in an epidemiological survey of the general population, between December 1984 and April 1987. Each participant was submitted to ultrasonography (US) of the gallbladder and completed a food-frequency questionnaire, covering 38 food items. A common portion size was identified and subjects were asked how often each item was consumed. Nutrient intake was computed by multiplying the intake frequency and nutrient content per portion for each item, and then by summing the product over all foods. Each nutrient intake was adjusted for energy intake. Alcohol intake was calculated by summing the consumption of wine, beer, and liquor. Having excluded subjects aware of having gallstones (GS) or previously submitted to cholecystectomy (to avoid prothopatic bias), 787 males and 1,014 females with GS and 14,272 males and 10,836 females without GS were available for analysis. Relative risks (RR) of GS were computed by quintiles of nutrient intake. The overnight fasting period was calculated as the difference between the specified time of dinner and the time of the next meal (breakfast or lunch). A significant negative association was found between RR of GS and total energy intake for males (chi2 for trend = 8.37; P = .004), fiber intake for females (chi2 = 5.45; P = .02), and daily alcohol consumption for males (chi2 = 10.86; P = .001). A positive association was observed between RR of GS and carbohydrate (chi2 = 5.95; P = .01 for males; chi2 = 9.39; P = .002 for females) and protein intake only for males (chi2 = 10.92; P = .01). Prevalence of GS was higher among subjects who had an overnight fasting period of over 12 hours than subjects with that of less than 12 hours. (RR: 1.35; 95% CI: 1.01-1.80 for males; RR: 1.28; 95% CI: 1.03-1.60 for females). These data do not confirm that high energy intake is associated with an increased risk of GS. Factors protecting against GS comprise: low carbohydrate (males and females) and protein (males) intakes, high fiber (females) and moderate alcohol intake (males) consumption, and a shorter overnight fasting period for both sexes.


Qualitative Parental Perceptions of a Paediatric Multidisciplinary Team Clinic for Prader-Willi Syndrome
This preliminary review was conducted to inform the design of a new service to support families with children with Prader-Willi syndrome (PWS). Families were invited to attend a pilot clinic at a hospital outpatient department, comprising appointments with a multi-disciplinary team (MDT). Following the clinic, families (n=6) were invited to take part in semi-structured qualitative interviews that were audio-recorded, transcribed and analysed using thematic analysis. Families reported that the clinic offered enhanced support in the following categories: integrated care; professional input; signposting to social support (respite and financial); connection with the wider PWS community; and behavioural support. This is the first paper that documents the parental perspective of an MDT clinic for children with PWS. The families felt an MDT clinic was superior to current care, offering more convenient access to an enhanced service, which would provide integrated and consistent care for their children's diverse, challenging and changing needs.


In a Case of Death Involving Steroids, Hair Testing is More Informative than Blood or Urine Testing.
A 59-year-old male was found dead at home, with two empty vials of an oily preparation obtained from a manufacturer from East Europe. There was no label on the vial. The subject was a former weightlifter, also known as an anabolic steroids abuser. The local prosecutor ordered a body examination, which was unremarkable, and allowed collecting femoral blood, urine and scalp hair (6 cm, brown). He was treated for cardiac insufficiency with quinidine. Biological specimens were submitted not only to standard toxicological analyses including a screening with liquid chromatography (LC)-quadrupole time of flight, but also to a specific LC-tandem mass spectrometry method for anabolic steroids testing. Ethanol was not found in both blood and urine. Quinidine blood concentration (791 ng/mL) was therapeutic. No drug of abuse was identified. In blood, testosterone was less that 1 ng/mL and no other steroid was identified. In urine, testosterone/epitestosterone was 1.56 and boldenone was present at a concentration of 9 ng/mL. The hair test results, performed on the whole length, demonstrated repetitive steroids abuse, including not only testosterone (140 pg/mg), testosterone propionate (605 pg/mg) and testosterone decanoate (249 pg/mg), but also boldenone (160 pg/mg), trenbolone (143 pg/mg) and metandienone (60 pg/mg). Since forensic laboratories have limited access to steroid urinary metabolite reference material due to specific regulations (to avoid testing athletes before anti-doping verifications), hair analyses seem to be the best approach to document anabolic agents abuse. Indeed, in hair, the target drug is the parent compound; in addition, when compared to blood or urine, this matrix has a much larger window of detection. The pathologist concluded cardiac insufficiency in a context involving repetitive abuse of anabolic drugs. This case indicates that more attention should be paid to anabolic steroids, in a context of sudden cardiac death.


Understanding the age divide in COVID-19: why are children overwhelmingly spared?
The rapid emergence and subsequent global dissemination of SARS-CoV-2 disease (COVID-19) has resulted in over 4 million cases worldwide. The disease has a marked predilection for adults, and children are relatively spared. Understanding the age-based differences in pathophysiological pathways and processes relevant to the onset and progression of disease both in the clinical course and in experimental disease models may hold the key to the identification of therapeutic targets. The differences in the clinical course are highlighted by the lack of progression of the SARS-CoV-2 infection beyond mild symptoms in a majority of children, whereas in adults the disease progresses to acute lung injury and an acute respiratory distress syndrome (ARDS)-like phenotype with high mortality. The pathophysiological mechanisms leading to decreased lung injury in children may involve the decreased expression of the mediators necessary for viral entry into the respiratory epithelium and differences in the immune system responses in children. Specifically, decreased expression of proteins, including angiotensin-converting enzyme 2 (ACE2) and Transmembrane Serine Protease 2 (TMPRSS2) in the airway epithelium in children may prevent viral entry. The immune system differences may include a relative preponderance of CD4+ T cells, decreased neutrophil infiltration, decreased production of proinflammatory cytokines, and increased production of immunomodulatory cytokines in children compared with adults. Notably, the developing lung in children may have a greater capacity to recover and repair after viral infection. Understanding the relative contributions of the above processes to the protective phenotype in the developing lung can guide the trial of the appropriate therapies in adults.


Neuropsychological evidence for a spatial bias in visual short-term memory after left posterior ventral damage.
For the first time, we report a spatial bias in visual short-term memory (VSTM) after left medial and inferior occipito-temporal damage. Our patient D.M. showed a spatial bias in report from VSTM, being more accurate at reporting stimuli presented in her left than her right visual field (Experiment 1). This spatial bias could not be attributed to a visual field deficit (Experiment 2) and was based on the relative rather than the absolute locations of the stimuli (Experiment 3). It was reduced when the transfer of items to VSTM was facilitated-for example, by grouping stimuli (Experiment 4) or by reducing the number of items to be remembered (Experiment 5). The spatial bias was attenuated when items moved from right to centre or left to centre, and D.M. was cued to report the item that would have been on the right or left, had the movement continued (Experiment 6). We conclude that posterior ventral damage can impair both the consolidation of new information in VSTM and the explicit report of consolidated information from VSTM.


Anesthesia and the porphyrias.
A simplified classification of the porphyrias is given which is thought to be advantageous to the anesthesiologist in determining those patients who are predisposed to acute attacks. These acute attacks may be precipitated by the administration of barbiturates, but may also be spontaneous. The current theory for the precipitation of the acute attack is described, with the probable mechanism being a decrease in uroporphyrinogen synthetase levels and the resultant interference in heme production. Increased formation of cytochrome P-450 with barbiturates also produces increased levels of delta aminolevulinic acid, which may be a cause of the acute attack. The significance in anesthesia and suggested means of anesthetic management are discussed.


Uncontrolled manifold hypothesis: Organization of leg joint variance in humans while walking in a wide range of speeds.
This study aimed at investigating the organization of joint angle variability during walking by using the uncontrolled manifold (UCM) theory. We tested two hypotheses: i. the coordinative mechanism underlying joint angle variance during the stance phase is compatible with a kinematic synergy that stabilizes the centre of mass (CoM) position; ii. the walking speed affects the variance components onto and orthogonal to the UCM. Eight healthy subjects (26.0±2.0years old) steadily walked on a treadmill at five normalised speeds (from 0.62±0.03m/s to 1.15±0.07m/s). Joint angles and foot orientation, and components of the CoM position were, respectively, used as elemental variables and task performance for the UCM implementation. The effect of speed, time events, and variance components on the distribution of data variance in the space of joint angles was analyzed by the ANOVA test. Results corroborated the hypothesis that the variance of elemental variables is structured in order to minimize the stride-to-stride variability of the CoM position, at all speeds. Noticeably, both variance components increase during the propulsive phase, albeit that parallel to the UCM was always grater than the orthogonal one. Accordingly, the observed kinematic synergy is supposed to contribute to accomplishing an efficient transition between two steps. Results also revealed that the walking speed does not affect the partitioning of elemental variables-related variance onto and orthogonal to the UCM. Accordingly, the organization of leg joint variance underlying the stabilization of CoM position remains almost unaltered across speeds.


Novel activating BRAF fusion identifies a recurrent alternative mechanism for ERK activation in pediatric Langerhans cell histiocytosis.
Langerhans cell histiocytosis (LCH) is an inflammatory myeloid neoplasm characterized by constitutive activation of extracellular signal-regulated kinase (ERK). Genomic characterization has identified activating point mutations including mutually exclusive BRAFV600E and activating MAP2K1 mutations to be responsible for ERK activation in a majority of pediatric LCH patients. Here, we report the discovery of a novel BRAF kinase fusion, PACSIN2-BRAF, in a child with multisystem LCH. This is the second reported case of an activating BRAF kinase fusion and indicates a recurrent pathologic mechanism. Genomic evaluation for activating kinase fusions should be strongly considered in pediatric LCH patients lacking more common mutations.


I must have missed that: Alpha-band oscillations track attention to spoken language.
Attention is critical to the construction of mental representations of language context during comprehension. We investigated the consequences of momentary lapses in attention during listening comprehension on neural activity and behavior. Participants listened to two full-length stories while EEG was recorded, and afterwards completed multiple choice comprehension questions. Listening was periodically interrupted by attention probes, in which participants were asked whether their attention immediately preceding the probe's appearance was focused on the story. The results showed that (1) participants spent a substantial amount of time off-task, endorsing attention lapses on over 30% of probes; (2) for probes on which an attention lapse was endorsed, later accuracy on comprehension questions querying pre-probe information was decreased; (3) the pre-probe period just before the endorsement of an attention lapse was characterized by a greater percentage of above-threshold oscillations in the alpha-band (8-12 Hz) compared to just prior to the endorsement of on-task or split-attention listening; and (4) when participants made "I have no idea" responses to comprehension questions, their EEG record revealed a greater percentage of above-threshold alpha oscillations during the original presentation of the information queried by the comprehension questions, compared to correct responses or incorrect guesses. These results connect changes in neural activity in the alpha band to episodes of mind-wandering during listening comprehension, and in turn to decreased comprehension accuracy. This demonstrates how alpha can be used to track attentional engagement during language comprehension, and illustrates the dependence of successful language comprehension on attention.


Predicting Outcome in Guillain-Barré Syndrome: International Validation of the Modified Erasmus GBS Outcome Score.
The clinical course and outcome of the Guillain-Barré syndrome (GBS) are diverse and vary among regions. The modified Erasmus GBS Outcome Score (mEGOS), developed with data from Dutch patients, is a clinical model that predicts the risk of walking inability in patients with GBS. The study objective was to validate the mEGOS in the International GBS Outcome Study (IGOS) cohort and to improve its performance and region specificity. We used prospective data from the first 1,500 patients included in IGOS, aged ≥6 years and unable to walk independently. We evaluated whether the mEGOS at entry and week 1 could predict the inability to walk unaided at 4 and 26 weeks in the full cohort and in regional subgroups, using 2 measures for model performance: (1) discrimination: area under the receiver operating characteristic curve (AUC) and (2) calibration: observed vs predicted probability of being unable to walk independently. To improve the model predictions, we recalibrated the model containing the overall mEGOS score, without changing the individual predictive factors. Finally, we assessed the predictive ability of the individual factors. For validation of mEGOS at entry, 809 patients were eligible (Europe/North America [n = 677], Asia [n = 76], other [n = 56]), and 671 for validation of mEGOS at week 1 (Europe/North America [n = 563], Asia [n = 65], other [n = 43]). AUC values were >0.7 in all regional subgroups. In the Europe/North America subgroup, observed outcomes were worse than predicted; in Asia, observed outcomes were better than predicted. Recalibration improved model accuracy and enabled the development of a region-specific version for Europe/North America (mEGOS-Eu/NA). Similar to the original mEGOS, severe limb weakness and higher age were the predominant predictors of poor outcome in the IGOS cohort. mEGOS is a validated tool to predict the inability to walk unaided at 4 and 26 weeks in patients with GBS, also in countries outside the Netherlands. We developed a region-specific version of mEGOS for patients from Europe/North America. This study provides Class II evidence that the mEGOS accurately predicts the inability to walk unaided at 4 and 26 weeks in patients with GBS. NCT01582763.


Impact of tobacco-pack pictorial warnings on youth and young adults: A systematic review of experimental studies.
We conducted a systematic review of the experimental literature on the impact of tobacco-pack pictorial warning labels (PWLs) on youth and young adults. We systematically searched computerized databases and the reference lists of relevant articles. We included studies that used an experimental protocol to assess PWLs. Studies had to report findings for youth or young adult samples (aged <30 years). Thirty-one studies met the inclusion criteria, with a total sample size of 27506. Two coders independently coded all study characteristics and outcomes. Twenty-eight studies experimentally evaluated PWLs for cigarette packs while three studies evaluated PWLs for smokeless tobacco packs. Generally, PWLs led to higher attention, stronger cognitive and affective reactions, more negative pack attitudes and smoking attitudes, and increased intentions not to use tobacco products compared to text warnings. PWLs were perceived to be more effective than text warnings for both cigarette packs and smokeless tobacco packs. The systematic review showed that PWLs on tobacco products are effective across a wide range of tobacco-related outcomes among young people. Gaps in the literature include a lack of research on tobacco initiation and cessation and a dearth of literature on non-cigarette tobacco products.


Eigenbehaviour as an Indicator of Cognitive Abilities.
With growing use of machine learning algorithms and big data in health applications, digital measures, such as digital biomarkers, have become highly relevant in digital health. In this paper, we focus on one important use case, the long-term continuous monitoring of cognitive ability in older adults. Cognitive ability is a factor both for long-term monitoring of people living alone as well as a relevant outcome in clinical studies. In this work, we propose a new potential digital biomarker for cognitive abilities based on location eigenbehaviour obtained from contactless ambient sensors. Indoor location information obtained from passive infrared sensors is used to build a location matrix covering several weeks of measurement. Based on the eigenvectors of this matrix, the reconstruction error is calculated for various numbers of used eigenvectors. The reconstruction error in turn is used to predict cognitive ability scores collected at baseline, using linear regression. Additionally, classification of normal versus pathological cognition level is performed using a support-vector machine. Prediction performance is strong for high levels of cognitive ability but grows weaker for low levels of cognitive ability. Classification into normal and older adults with mild cognitive impairment, using age and the reconstruction error, shows high discriminative performance with an ROC AUC of 0.94. This is an improvement of 0.08 as compared with a classification with age only. Due to the unobtrusive method of measurement, this potential digital biomarker of cognitive ability can be obtained entirely unobtrusively-it does not impose any patient burden. In conclusion, the usage of the reconstruction error is a strong potential digital biomarker for binary classification and, to a lesser extent, for more detailed prediction of inter-individual differences in cognition.


Concise review: microfluidic technology platforms: poised to accelerate development and translation of stem cell-derived therapies.
Stem cells are a powerful resource for producing a variety of cell types with utility in clinically associated applications, including preclinical drug screening and development, disease and developmental modeling, and regenerative medicine. Regardless of the type of stem cell, substantial barriers to clinical translation still exist and must be overcome to realize full clinical potential. These barriers span processes including cell isolation, expansion, and differentiation; purification, quality control, and therapeutic efficacy and safety; and the economic viability of bioprocesses for production of functional cell products. Microfluidic systems have been developed for a myriad of biological applications and have the intrinsic capability of controlling and interrogating the cellular microenvironment with unrivalled precision; therefore, they have particular relevance to overcoming such barriers to translation. Development of microfluidic technologies increasingly utilizes stem cells, addresses stem cell-relevant biological phenomena, and aligns capabilities with translational challenges and goals. In this concise review, we describe how microfluidic technologies can contribute to the translation of stem cell research outcomes, and we provide an update on innovative research efforts in this area. This timely convergence of stem cell translational challenges and microfluidic capabilities means that there is now an opportunity for both disciplines to benefit from increased interaction.


A novel combined ex vivo and in vivo lentiviral interleukin-10 gene delivery strategy at the time of transplantation decreases chronic lung allograft rejection in mice.
Our objective was to develop a rapid-onset and durable gene-delivery strategy that is applicable at the time of transplant to determine its effects on both acute rejection and chronic lung allograft fibrosis using a mouse orthotopic lung transplant model. C57BL/6 mice received an orthotopic left lung transplant from syngeneic donors or C57BL/10 donors. By using syngeneic lung transplantation, we established a novel gene transfer protocol with lentivirus luciferase intrabronchial administration to the donor lung ex vivo before transplantation. This strategy was applied in allogeneic lung transplantation with lentivirus engineering expression of human interleukin-10 or lentivirus luciferase (control). Bioluminescent imaging revealed that the highest early transgene expression was achieved when lentivirus luciferase was administered both directly into the donor lung graft ex vivo before implantation and subsequently to the recipient in vivo daily on post-transplant days 1 to 4, compared with post-transplant in vivo administration only (days 0 to 4). Our previous work with adenoviral interleukin-10 gene therapy indicates that early interleukin-10 expression in the allograft is desirable. Therefore, we selected the combined protocol for human interleukin-10 encoding lentiviral vector therapy. In the allogeneic transplant setting, ex vivo and in vivo human interleukin-10 encoding lentiviral vector therapy reduced acute rejection grade (2.0 vs 3.0, P < .05) at day 28. The percentage of fibrotic obliterated airways was reduced in the human interleukin-10 encoding lentiviral vector-treated group (10.9% ± 7.7% vs 40.9% ± 9.3%, P < .05). Delivery of lentiviral interleukin-10 gene therapy, using a novel combined ex vivo and in vivo delivery strategy, significantly improves acute and chronic rejection in the mouse lung transplant model.


Role of persisters and small-colony variants in antibiotic resistance of planktonic and biofilm-associated Staphylococcus aureus: an in vitro study.
The presence of persister cells and small-colony variants (SCVs) has been associated with enhanced antibiotic resistance of many organisms in biofilms. This study investigated whether persisters and/or SCVs contribute to the antibiotic resistance of Staphylococcus aureus biofilms. A detailed dose-dependent killing of biofilms and planktonic cells with five antibiotics (oxacillin, cefotaxime, amikacin, ciprofloxacin and vancomycin) was analysed by treating them with each antibiotic at a concentration of 0-100 microg ml(-1) at 37 degrees C for 48 h. The killing of biofilm cells by all of the antibiotics showed the presence of persister cells - most cells in the population died, leaving a fraction that persisted, even at higher concentrations of the antibiotics. These persisters represented a transient resistant phenotype and reverted to a killing curve resembling that of the wild-type parent upon re-exposure to the antibiotics. SCVs were observed in biofilms only after treatment with ciprofloxacin, and these SCVs were of a transient nature. The treatment of planktonic cells with oxacillin, cefotaxime, ciprofloxacin and vancomycin killed the entire population and no persisters were detected. Transient SCVs, observed in planktonic cells following exposure to these antibiotics, were killed at higher antibiotic concentrations. The treatment of planktonic cells with amikacin yielded a small subpopulation of survivors that included persisters (at numbers significantly lower than for the biofilms) and highly resistant, stable SCVs with an increased biofilm-forming capacity in comparison with the wild-type parent. Thus the high resistance of S. aureus biofilms to multiple unrelated antibiotics is largely dependent on the presence of persister cells. Biofilms harbour a large number of persisters in comparison with planktonic cultures, which either do not harbour persisters or harbour only a small number. SCVs, although not specifically associated with S. aureus biofilms, have an increased biofilm-forming capacity and this may explain the frequent isolation of SCVs from biofilm-associated infections. The intrinsic resistance of these variants may in turn contribute to the enhanced antibiotic resistance of the biofilms thus formed.


Power-law spatial correlations in arrays of locally coupled lasers.
We investigate correlations of the intensity fluctuations of two-dimensional arrays of nonidentical, locally coupled lasers, numerically and experimentally. We find evidence of a power-law dependence of spatial correlations as a function of laser pair distance (or coupling strength) near the phase-locking threshold.


Energetics and diffusion of liquid water and hydrated ions through nanopores in graphene: ab initio molecular dynamics simulation.
The energetics and diffusion of water molecules and hydrated ions (Na+, Cl-) passing through nanopores in graphene are addressed by dispersion-corrected density functional theory calculations and ab initio molecular dynamics (MD) simulations. Pores of about 0.8 nm in diameter with different pore-edge passivations, with (H) and (O, H) atoms, were considered. Our MD simulations show a water flux through the hydroxylated pores of about one H2O molecule every three picoseconds, in close agreement with recent experiments that estimated a water flux of three molecules per picosecond through pores of ∼1 nm. We also find that both pores are effective in blocking hydrated Na+ and Cl- ions with large energy barriers, ranging from 12 to 15 eV. In addition, pore passivation with O atoms would increase the water transport through hydroxylated pores, due to the formation of hydrogen bonds with nearby water molecules, which is not observed in the hydrogenated pores.


Cigarette smoke constituents cause endothelial nitric oxide synthase dysfunction and uncoupling due to depletion of tetrahydrobiopterin with degradation of GTP cyclohydrolase.
Cigarette smoking (CS) is a well-established risk factor for cardiovascular disease (CVD). Endothelial dysfunction (ED) with loss of nitric oxide (NO) production is a central mechanism leading to the advent of CVD. Despite many prior studies of this major health problem, the exact mechanism by which CS induces ED is not well understood. This study examines the mechanism by which CS induces ED with altered endothelial NO synthase (eNOS) function in aortic endothelial cells (AECs). Exposure of AECs to cigarette smoke extract (CSE) resulted in a marked decrease in NO production with concomitant increase in superoxide (O2.-) generation and accumulation of 4-hydroxy-2-nonenal protein adducts. CSE exposure led to depletion of the essential eNOS cofactor tetrahydrobiopterin (BH4) as well as total biopterin levels and decreased the expression level of guanosine triphosphate cyclohydrolase (GTPCH), the rate limiting enzyme in BH4 biosynthesis. Moreover, exposure of AECs to CSE increased the level of ubiquitinated proteins and increased 26 S proteasomal activity in a concentration-dependent manner. Pre-treatment with MG132, a 26 S proteasome inhibitor, partially prevented CSE-induced loss of BH4, total biopterin, GTPCH, and increased NO production following CSE exposure, indicating a role of the ubiquitin-proteasome system in CSE-induced eNOS dysfunction. In conclusion, CSE-induced eNOS dysfunction and uncoupling occurs due to BH4 depletion with BH4de novo synthesis limited by diminished GTPCH expression.


[Ethanol for treatment of delirium in alcohol dependent patients on intensive care units in the Netherlands: efficacy not proven].
To determine the use of ethanol in Dutch intensive care departments (ICUs) for the treatment of deliriant symptoms in alcohol-dependent patients, and to study the literature data concerning this use. Literature search and questionnaire. In Medline a search was performed from 1993 onward with keywords 'intensive care' and 'alcohol'/'alcohol withdrawal syndrome'/'delirium'. All Dutch ICUs received a written questionnaire (n = 247) concerning the use of ethanol in this patient group. According to the literature the syndrome is characterized by autonomous hyperactivity, resulting in tachycardia, tachypnoea, hypertension, perspiration, fever, tremors en fear. In delirium caused by alcohol withdrawal benzodiazepines are advised, sometimes in combination with haloperidol. ICUs sometimes use ethanol, although the effectiveness in preventing or treating withdrawal symptoms has never been ascertained in scientific investigations: in a meta-analysis the conclusion is drawn that the studies are too small and insufficiently objective to determine the effectiveness. The response to the questionnaire was 55% (96/176). From all ICUs, 15 (16%) used ethanol occasionally, and treated an estimated 17% of the admitted alcohol-dependent patients. There was a tendency for large ICUs to use ethanol in these patients more frequently. Each ICU used its own method to calculate the required ethanol dosage. Calculation on the basis of these dosages indicated a plasma ethanol concentration of at most 0.5 promille. Because only a small percentage of deliriums on ICUs are caused by alcohol withdrawal, and the effectiveness of ethanol in alcohol-dependent patients with a delirium has never been proven, the use of ethanol in such cases is discouraged.


Outcomes of hospitalized COVID-19 patients in Canada: impact of ethnicity, migration status and country of birth.
Ethnoracial groups in high-income countries have a 2-fold higher risk of SARS-CoV-2 infection, associated hospitalizations, and mortality than Whites. Migrants are an ethnoracial subset that may have worse COVID-19 outcomes due to additional barriers accessing care, but there are limited data on in-hospital outcomes. We aimed to disaggregate and compare COVID-19 associated hospital outcomes by ethnicity, immigrant status and region of birth. Adults with community-acquired SARS-CoV-2 infection, hospitalized March 1-June 30, 2020, at four hospitals in Montréal, Quebec, Canada, were included. Age, sex, socioeconomic status, comorbidities, migration status, region of birth, self-identified ethnicity [White, Black, Asian, Latino, Middle East/North African], intensive care unit (ICU) admissions and mortality were collected. Adjusted hazard ratios (aHR) for ICU admission and mortality by immigrant status, ethnicity and region of birth adjusted for age, sex, socioeconomic status and comorbidities were estimated using Fine and Gray competing risk models. Of 1104 patients (median [IQR] age, 63.0 [51.0-76.0] years; 56% males), 57% were immigrants and 54% were White. Immigrants were slightly younger (62 vs 65 years; p = 0.050), had fewer comorbidities (1.0 vs 1.2; p < 0.001), similar crude ICU admissions rates (33.0% vs 28.2%) and lower mortality (13.3% vs 17.6%; p < 0.001) than Canadian-born. In adjusted models, Blacks (aHR 1.39, 95% confidence interval 1.05-1.83) and Asians (1.64, 1.15-2.34) were at higher risk of ICU admission than Whites, but there was significant heterogeneity within ethnic groups. Asians from Eastern Asia/Pacific (2.15, 1.42-3.24) but not Southern Asia (0.97, 0.49-1.93) and Caribbean Blacks (1.39, 1.02-1.89) but not SSA Blacks (1.37, 0.86-2.18) had a higher risk of ICU admission. Blacks had a higher risk of mortality (aHR 1.56, p = 0.049). Data disaggregated by region of birth identified subgroups of immigrants at increased risk of COVID-19 ICU admission, providing more actionable data for health policymakers to address health inequities.


Drownings of children and youth in a desert state.
Despite the fact that New Mexico is a desert state, the drowning rate (4.3) is 40% higher than that nationally (3.0). More than half of those who drown are younger than 25 years, toddlers and adolescents being at greater risk than other age groups. In an epidemiologic study of a six-year period, male drowning victims outnumbered female and American Indian and Hispanic victims outnumbered whites. Other major risk factors were epilepsy, mental retardation and alcohol abuse. In most drownings in the under-25 age group, the victim was involved in recreation near the water or swimming, although many were partially or fully clothed. Most drownings occurred in lakes, ditches or rivers. Because New Mexico is not a water-oriented state, public awareness of drowning hazards needs to be increased and more intensive preventive efforts instituted.


Targeted genes and interacting proteins of hypoxia inducible factor-1.
Heterodimeric transcription factor hypoxia inducible factor-1 (HIF-1) functions as a master regulator of oxygen homeostasis in almost all nucleated mammalian cells. The fundamental process adapted to cellular oxygen alteration largely depends on the refined regulation on its alpha subunit, HIF-1α. Recent studies have unraveled expanding and critical roles of HIF-1α, involving in a multitude of developmental, physiological, and pathophysiological processes. This review will focus on the current knowledge of HIF-1α-targeting genes and its interacting proteins, as well as the concomitant functional relationships between them.


Cigarette Smoke Extract Modulates Functions of Peroxisome Proliferator-Activated Receptors.
Cigarette smoke extract (CSE) contains many toxicants and may derange the physiological processes, such as cholesterol metabolism. We examined the impact of CSE on transcriptional regulation mediated peroxisome proliferator-activated receptors (PPARs) and its interaction with cofactors to elucidate differences in the molecular mechanism between CSE and other agonists of PPARs. We constructed several mutant PPARs (mPPARs) with amino acid substitution in the ligand-binding domain, which according to the molecular modeling, may affect the binding of agonists. In transient expression assays, each wild-type peroxisome proliferator-activated receptor (PPAR) mediated transcription stimulated by CSE was faintly yet significantly elevated compared to the control. The CSE-induced transcriptional activation was abolished in the H323A, H323Y, S342A, and H449A mPPARγs, although the activation elevated by pioglitazone was reserved. In the mPPARγ with Y473A and mPPARβ/δs with H286Y and Y436A, the pioglitazone-induced or L165041-activated transcriptional elevations were decreased and were lower than that of CSE-induced stimulation. These results suggested that CSE activated both mutant PPARs to be selectively different from those ligands. Mammalian two-hybrid assay illustrated that CSE could mildly recruit SRC1 or GRIP1 to the wild-type PPARγ. Representative ingredients, such as acrolein and crotonaldehyde present in CSE, could stimulate PPAR isoforms even at the toxicological concentrations and might possibly contribute to stimulatory effects. CSE mildly regulates the cholesterol metabolism-related genes, such as low density lipoprotein (LDL) receptor and Liver X receptor (LXR)β. In conclusion, these CSE effects the nuclear hormone receptors and their cofactors thereby disturbing metabolic phenomena. Therefore, CSE might be involved in cholesterol metabolism.


Effects of changes in mucosal solution Cl- or K+ concentration on cell water volume of Necturus gallbladder epithelium.
An electrophysiologic technique was used to measure changes in cell water volume in response to isosmotic luminal solution ion replacement. Intracellular Cl- activity (aCl-i) was measured and net flux determined from the changes in volume and activity. Reduction of luminal solution [Cl-] from 98 to 10 mM (Cl- replaced with cyclamate) resulted in a large fall in aCl-i with no significant change in cell water volume. Elevation of luminal solution [K+] from 2.5 to 83.5 mM (K+ replaced Na+) caused a small increase in aCl-i with no change in cell water volume. Exposure of the Necturus gallbladder epithelium to agents that increase intracellular cAMP levels (forskolin and/or theophylline) induces an apical membrane electrodiffusive Cl- permeability accompanied by a fall in aCl-i and cell shrinkage. In stimulated tissues, reduction of luminal solution [Cl-] resulted in a large fall in aCl-i and rapid cell shrinkage, whereas elevation of luminal solution [K+] caused a large, rapid cell swelling with no significant change in aCl-i. The changes in cell water volume of stimulated tissues elicited by lowering luminal solution [Cl-] or by elevating luminal solution [K+] were reduced by 60 and 70%, respectively, by addition of tetraethylammonium (TEA+) to the luminal bathing solution. From these results, we conclude that: (a) In control tissues, the fall in aCl-i upon reducing luminal solution [Cl-], without concomitant cell shrinkage, indicates that the Cl- entry mechanism is electroneutral (Cl-/HCO3-) exchange. (b) Also in control tissues, the small increase in aCl-i upon elevating luminal solution [K+] is consistent with the recent demonstration of a basolateral Cl- conductance. (c) The cell shrinkage elicited by elevation of intracellular cAMP levels results from conductive loss of Cl- (and probably K+). (d) Elevation of cAMP inhibits apical membrane Cl-/HCO-3-exchange activity by 70%. (e) The cell shrinkage in response to the reduction of mucosal solution [Cl-] in stimulated tissues results from net K+ and Cl- efflux via parallel electrodiffusive pathways. (f) A major fraction of the K+ flux is via a TEA(+)-sensitive apical membrane K+ channel.


Proteomic analysis of the skeletal muscles from dysferlinopathy patients.
Dysferlinopathy is an autosomal recessive disease caused by pathogenic variants in DYSF gene. We compared muscle protein extracts from dysferlinopathy patients and control subjects to identify new biomarkers of this myopathy. We reviewed the medical records from January 2002 to October 2016. Eight vastus lateralis muscle samples from five dysferlinopathy patients and three control subjects were selected. We separated proteins/peptides from all eight muscle protein extracts using two-dimensional electrophoresis (2DE). Data were acquired from liquid chromatography-mass spectrometry protein fragmentation patterns after comparing the spot volumes. Western blotting revealed total dysferlin loss in the dysferlinopathy patients but normal expression in the control subjects. 2DE indicated somewhat diverse protein constellations between the dysferlinopathy and control groups. Image analysis showed that 80 spots were differently expressed between two dysferlinopathy and one control samples. We selected 44 spots with consistently different volume between dysferlinopathy and control groups. Liquid chromatography-mass spectrometry indicated 26 differently expressed proteins. Western blotting revealed that creatine kinase M-type, carbonic anhydrase III (muscle specific) and desmin were significantly elevated in dysferlinopathy muscle. Additionally, four proteins (myosin light chain 1/3, skeletal muscle isoform; lamin A/C; ankyrin repeat domain 2; and eukaryotic translation initiation factor 5A-1) were inconsistently elevated in the dysferlinopathy samples. We confirmed the usefulness of the classic biomarker and have newly identified the altered expression of proteins in the skeletal muscles of dysferlinopathy patients.