High-throughput screening of the ReFRAME, Pandemic Box, and COVID Box drug repurposing libraries against SARS-CoV-2 nsp15 endoribonuclease to identify small-molecule inhibitors of viral activity.
SARS-CoV-2 has caused a global pandemic, and has taken over 1.7 million lives as of mid-December, 2020. Although great progress has been made in the development of effective countermeasures, with several pharmaceutical companies approved or poised to deliver vaccines to market, there is still an unmet need of essential antiviral drugs with therapeutic impact for the treatment of moderate-to-severe COVID-19. Towards this goal, a high-throughput assay was used to screen SARS-CoV-2 nsp15 uracil-dependent endonuclease (endoU) function against 13 thousand compounds from drug and lead repurposing compound libraries. While over 80% of initial hit compounds were pan-assay inhibitory compounds, three hits were confirmed as nsp15 endoU inhibitors in the 1-20 μM range in vitro. Furthermore, Exebryl-1, a ß-amyloid anti-aggregation molecule for Alzheimer's therapy, was shown to have antiviral activity between 10 to 66 μM, in Vero 76, Caco-2, and Calu-3 cells. Although the inhibitory concentrations determined for Exebryl-1 exceed those recommended for therapeutic intervention, our findings show great promise for further optimization of Exebryl-1 as an nsp15 endoU inhibitor and as a SARS-CoV-2 antiviral.


In vitro characterization of cell-level neurophysiological diversity in the rostral nucleus reuniens of adult mice.
The nucleus reuniens (Re), a nucleus of the midline thalamus, is part of a cognitive network including the hippocampus and the medial prefrontal cortex. To date, very few studies have examined the electrophysiological properties of Re neurons at a cellular level. The majority of Re neurons exhibit spontaneous action potential firing at rest. This is independent of classical amino-acid mediated synaptic transmission. When driven by various forms of depolarizing current stimulus, Re neurons display considerable diversity in their firing patterns. As a result of the presence of a low threshold Ca2+ channel, spike output functions are strongly modulated by the prestimulus membrane potential. Finally, we describe a novel form of activity-dependant intrinsic plasticity that eliminates the high-frequency burst firing present in many Re neurons. These results provide a comprehensive summary of the intrinsic electrophysiological properties of Re neurons allowing us to better consider the role of the Re in cognitive processes. The nucleus reuniens (Re) is the largest of the midline thalamic nuclei. We have performed a detailed neurophysiological characterization of neurons in the rostral Re of brain slices prepared from adult male mice. At resting potential (-63.7 ± 0.6 mV), ∼90% of Re neurons fired action potentials, typically continuously at ∼8 Hz. Although Re neurons experience a significant spontaneous barrage of fast, amino-acid-mediate synaptic transmission, this was not predominantly responsible for spontaneous spiking because firing persisted in the presence of glutamate and GABA receptor antagonists. With resting potential preset to -80 mV, -20 pA current injections revealed a mean input resistance of 615 MΩ and a mean time constant of 38 ms. Following cessation of this stimulus, a significant rebound potential was seen that was sometimes sufficiently large to trigger a short burst of very high frequency (100-300 Hz) firing. In most cells, short (2 ms), strong (2 nA) current injections elicited a single spike followed by a large afterdepolarizing potential which, when suprathreshold, generated high-frequency spiking. Similarly, in the majority of cells preset at -80 mV, 500 ms depolarizing current injections to cells led to a brief initial burst of very high-frequency firing, although this was lost when cells were preset at -72 mV. Biophysical and pharmacological experiments indicate a prominent role for T-type Ca2+ channels in the high-frequency bursting of Re neurons. Finally, we describe a novel form of activity-dependent intrinsic plasticity that persistently eliminates the burst firing potential of Re neurons.


Tissue factor activity in leukemia cells. Special reference to disseminated intravascular coagulation.
Tissue factor activity (TFA) of 10(8) leukemia cells was measured in 82 patients with acute nonlymphoid leukemia by the clotting method. The TFA bore a significant correlation to the development of disseminated intravascular coagulation (DIC) in these cases. Mean TFA value with standard deviation (SD) was 8.3 +/- 6.3 U in 48 cases with DIC, which was significantly higher than 0.3 +/- 4.2 U in 34 cases without DIC. Whereas Mean TFA in non-M3 was 0.9 +/- 6.3 U which was significantly lower than 37.2 +/- 2.3 U in M3, some non-M3 showed TFA as high as M3 and were complicated by DIC. In heparin treatment, dosage of heparin could not be controlled by either APTT or AcCT but was controlled by the extent of TFA of leukemia cells. Retrospective analysis of clinical features revealed that 97000X + 9000 units/day (X = logarithm value of TFA) of heparin is an adequate dosage for the successful treatment of DIC when TFA of leukemia cells is 0.8 U or more.


Micro (nano) plastic pollution: The ecological influence on soil-plant system and human health.
Global plastic pollution has been a serious problem since many years and micro (nano) plastics (MNPs) have gained attention from researchers around the world. This is because MNPs able to exhibit toxicology and interact with potentially toxic elements (PTEs) in the environment, causing soil toxicity. The influences of MNPs on the soil systems and plant crops have been overlooked despite that MNPs can accumulate in the plant root system and generate detrimental impacts to the terrestrial environments. The consumption of these MNPs-contaminated plants or fruits by humans and animals will eventually lead to health deterioration. The identification and measurement of MNPs in various soil samples is challenging, making the understanding of the fate, environmental and ecological of MNPs in terrestrial ecosystem is limited. Prior to sample assessment, it is necessary to isolate the plastic particles from the environment samples, concentrate the plastic particles for analysis purpose to meet detection limit for analytical instrument. The isolation and pre-concentrated steps are challenging and may cause sample loss. Herein, this article reviews MNPs, including their fate in the environment and toxic effects exhibited towards soil microorganisms, plants and humans along with the interaction of MNPs with PTEs. In addition, various analysis methods of MNPs and management of MNPs as well as the crucial challenges and future research studies in combating MNPs in soil system are also discussed.


[How can we nowadays select the best embryo to transfer?].
Multiple pregnancies stand as the most common adverse outcome of assisted reproduction technologies (ART) and the dangers associated with those pregnancies have been reduced by doing elective single embryo transfers (e-SET). Many studies have shown that e-SET is compatible with a continuously high pregnancy rate per embryo transfer. Yet, it still becomes necessary to improve the selection process in order to define the quality of individual embryos - so that the ones we choose for transfer are more likely to implant. First, analysis of embryo morphology has greatly helped in this identification and remains the most relevant criterion for choosing the embryo. The introduction of time-lapse imaging provides new criteria predictive of implantation potential, but the real contribution of this system - including the benefit/cost ratio - seems to be not yet properly established. In this context, extended culture until blastocyst stage is an essential practice but it appears wise to keep it for a population showing a good prognosis. Then, the failure of aneuploid embryos to implant properly led to achieve preimplantation genetic screening (PGS) in order to increase pregnancy and delivery rates after ART. However, PGS by fluorescence in situ hybridization (FISH) at day 3 is a useless process - and may even be harmful. Another solution involves using comparative genomic hybridisation (CGH) and moving to blastocyst biopsy. Finally, it is envisaged that morphology will also be significantly aided by non-invasive analysis of biomarkers in the culture media that give a better reflection of whole-embryo physiology and function.


Diversity in the Emerging Critical Care Workforce: Analysis of Demographic Trends in Critical Care Fellows From 2004 to 2014.
Diversity in the physician workforce is essential to providing culturally effective care. In critical care, despite the high stakes and frequency with which cultural concerns arise, it is unknown whether physician diversity reflects that of critically ill patients. We sought to characterize demographic trends in critical care fellows, who represent the emerging intensivist workforce. We used published data to create logistic regression models comparing annual trends in the representation of women and racial/ethnic groups across critical care fellowship types. United States Accreditation Council on Graduate Medical Education-approved residency and fellowship training programs. Residents and fellows employed by Accreditation Council on Graduate Medical Education-accredited training programs from 2004 to 2014. None. From 2004 to 2014, the number of critical care fellows increased annually, up 54.1% from 1,606 in 2004-2005 to 2,475 in 2013-2014. The proportion of female critical care fellows increased from 29.5% (2004-2005) to 38.3% (2013-2014) (p < 0.001). The absolute number of black fellows increased each year but the percentage change was not statistically significantly different (5.1% in 2004-2005 vs 3.9% in 2013-2014; p = 0.92). Hispanic fellows increased in number from 124 (7.7%) in 2004-2005 to 216 (8.4%) in 2013-2014 (p = 0.015). The number of American Indian/Alaskan Native/Native Hawaiian/Pacific Islander fellows decreased from 15 (1.0%) to seven (0.3%) (p < 0.001). When compared with population estimates, female critical care fellows and those from racial/ethnic minorities were underrepresented in all years. The demographics of the emerging critical care physician workforce reflect underrepresentation of women and racial/ethnic minorities. Trends highlight increases in women and Hispanics and stable or decreasing representation of non-Hispanic underrepresented minority critical care fellows. Further research is needed to elucidate the reasons underlying persistent underrepresentation of racial and ethnic minorities in critical care fellowship programs.


Another potential source of error in Rh-hr typing.
Three cases are presented in which the presence of sodium caprylate dependent albumin auto-agglutinins in the patients' sera resulted in erroneous Rh-hr phenotyping of their erythrocytes. In the initial testing, the albumin control was negative in each case. Since one of the cases was an Rh0(D) negative individual whose red blood cells might have been classified as Rh0(D) positive, a study was made using routine Rh-hr typing reagents from different manufacturers. Attention is drawn to the fact that Rh-hr typings should be performed using washed red blood cell samples when serum is known to contain albumin autoagglutinins.


N,N'bis-(2-mercaptoethyl) isophthalamide (NBMI) exerts neuroprotection against lead-induced toxicity in U-87 MG cells.
N,N'-bis(2-mercaptoethyl)isophthalamide (NBMI) is a novel lipophilic heavy metal chelator and thiol redox antioxidant. This study was designed to investigate the neuroprotective activity of NBMI in U-87 MG cells exposed to lead acetate (PbAc). Cells were pretreated with NBMI for 24 h prior to a 48 h exposure to PbAc. Cell death (55%, p < 0.0001) and reduction of intracellular GSH levels (0.70-fold, p < 0.005) induced by 250 µM Pb were successfully attenuated by NBMI pretreatment at concentrations as low as 10 µM. A similar pretreatment with the FDA-approved Pb chelator dimercaptosuccinic acid (DMSA) proved ineffective, indicating a superior PKPD profile for NBMI. Pretreatment with NBMI successfully counteracted Pb-induced neuroinflammation by reducing IL-1β (0.59-fold, p < 0.05) and GFAP expression levels. NBMI alone was also found to significantly increase ferroportin expression (1.97-fold, p < 0.05) thereby enhancing cellular ability to efflux heavy metals. While no response was observed on the apoptotic pathway, this study demonstrated for the first time that necrotic cell death induced by Pb in U-87 MG cells is successfully attenuated by NBMI. Collectively these data demonstrate NBMI to be a promising neuroprotective compound in the realm of Pb poisoning.


Factors associated with hemoglobin glycation index in adults with type 1 diabetes mellitus: The FGM-Japan study.
The discrepancy between HbA1c and glucose exposure may have significant clinical implications; however, the association between the hemoglobin glycation index (HGI) and clinical parameters in type 1 diabetes remains controversial. This study aimed to find the factors associated with HGI (laboratory HbA1c - predicted HbA1c derived from the continuous glucose monitoring [CGM]). We conducted a cross-sectional study of adults with type 1 diabetes (n = 211, age 50.9 ± 15.2 years old, female sex = 59.2%, duration of CGM use = 2.1 ± 1.0 years). All subjects wore the CGM for 90 days before HbA1c measurement. Data derived from the FreeStyle Libre sensor were used to calculate the glucose management indicator (GMI) and glycemic variability (GV) parameters. HGI was defined as the difference between the GMI and the laboratory HbA1c levels. The participants were divided into three groups according to the HGI tertile (low, moderate, and high). Multivariate regression analyses were performed. The female sex ratio, HbA1c, and % coefficient of variation (%CV) significantly increased over the HGI tertile, while eGFR and Hb decreased over the HGI tertile. In multivariate analysis, the factors associated with HGI were %CV and eGFR, after adjusting for HbA1c level and sex (R2 = 0.44). This study demonstrated that HGI is associated with female sex, eGFR, and some glycemic variability indices, independently of HbA1c. Minimizing glycemic fluctuations might reduce HGI. This information provides diabetic health professionals and patients with personalized diabetes management for adults with type 1 diabetes.


Gene expression profiling in the study of the pathogenesis of systemic lupus erythematosus.
Systemic lupus erythematosus (SLE) is a systemic autoimmune disease with a complex pathogenesis involving multiple genetic and environmental contributions. DNA microarray technology has recently been applied to unravel some of this complexity through genomewide profiling. Early studies using microarray analysis of peripheral blood mononuclear cells (PBMCs) from SLE patients revealed dysregulation of inflammatory cytokines, chemokines, and immune response-related genes, as well as genes involved in apoptosis, signal transduction, and the cell cycle. More recently, using arrays containing many more genes, 4 independent research groups have found that interferon (IFN)-regulated genes are highly overexpressed in the peripheral blood and kidney glomeruli of SLE patients, supporting a crucial role for interferon in SLE. Future studies focusing on target tissues or organs in lupus, including the kidney, may further contribute to our understanding of lupus pathogenesis while providing new targets for therapy.


Effects of CGP 28014 on the in vivo release and metabolism of dopamine in the rat striatum assessed by brain microdialysis.
The effects on rat striatal dopamine (DA) metabolism of systemic and local administration of CGP 28014, an inhibitor of catechol-O-methyl-transferase (COMT), were studied by in vivo microdialysis. CGP 28014 (30 mg/kg i.p.) significantly reduced the levels of homovanillic acid (HVA), but did not modify DA and 3,4-dihydroxyphenylacetic acid (DOPAC). The intrastriatal administration (via the microdialysis probe) of 5, 7.5, 10, and 20 mM of CGP 28014 elicited a concentration-dependent, several-fold increase in extracellular DA but did not alter the levels of HVA and DOPAC. Thus, the effects of CGP 28014 observed after i.p. injection (decrease in HVA levels) are different from those measured after intrastriatal administration (increase in DA release). Therefore, the inhibition of COMT is likely to be due to the action of a metabolite of CGP 28014 formed in the periphery and not in the brain.


Difficulties in the prospective diagnosis of functional adrenal diseases by CT.
The accuracy of prospective diagnosis of functional adrenal disorders by computed tomography (CT) was evaluated in 65 patients strongly suspected of having such disorders. Thirty-seven patients also underwent adrenal venography and selective adrenal venous sampling. Of the 65 patients, 21 had a hyperfunctioning cortical adenoma, 25 had cortical or medullary hyperplasia, and 11 pheochromocytoma, while 10 had no functional adrenal disease. Diagnosis by CT alone was correct in 66%; diagnosis by CT and venous sampling was correct in 89%. Incorrect prospective CT diagnosis resulted most often from a normal appearance of functionally hyperplastic glands (9 cases), nodular hyperplasia simulating a focal adenoma (8 cases), or an incidental nonfunctioning mass (3 cases). This study emphasizes the need to correlate CT findings with biochemical evaluation and the usefulness of venous sampling in selected cases to avoid inappropriate surgery. When CT is interpreted in correlation with complete biochemical analysis, correct diagnosis can usually be made. A small number of cases may still require adrenal venous sampling.


Alcohol consumption--a risk factor for hemorrhagic and non-hemorrhagic stroke.
The risks of alcohol consumption and its association with stroke were studied in 621 patients with stroke and 573 control subjects using case-control methods. Patients with stroke were subdivided into 193 with subarachnoid hemorrhage, 91 with intracerebral hemorrhage, and 337 with cerebral infarction. Data on recent alcohol consumption were obtained by questionnaire in patients with stroke and compared with data from an occupational screening survey in control subjects. Relative risks, adjusted for confounding variables, exhibited J-shaped associations with increasing levels of alcohol consumption classified into four categories--abstainer, 1 to 90 g, 100 to 390 g, and greater than or equal to 400 g weekly). The individual risks were 1, 0.7, 0.5, and 1.3 for subarachnoid hemorrhage; 1.0, 0.6, 0.5., and 2.5 for intracerebral hemorrhage, and 1.0, 0.6, 0.7, and 2.4 for cerebral infarction for men and women combined. The results suggest that low levels of alcohol consumption may have some protective effect upon the cerebral vasculature, whereas heavy consumption predisposes to both hemorrhagic and non-hemorrhagic stroke.


Rightward biases in free-viewing visual bisection tasks: implications for leftward responses biases on similar tasks.
Neurologically normal individuals exhibit strong leftward response biases during free-viewing perceptual judgments of brightness, quantity, and size. When participants view two mirror-reversed objects and they are forced to choose which object appears darker, more numerous, or larger, the stimulus with the relevant feature on the left side is chosen 60-75% of the time. This effect could be influenced by inaccurate judgments of the true centre-point of the objects being compared. In order to test this possibility, 10 participants completed three visual bisection tasks on stimuli known to elicit strong leftward response biases. Participants were monitored using a remote eye-tracking device and instructed to stare at the subjective midpoint of objects presented on a computer screen. Although it was predicted that bisection errors would deviate to the left of centre (as is the case in the line bisection literature), the opposite effect was found. Significant rightward bisection errors were evident on two of the three tasks, and the leftward biases seen during forced-choice tasks could be the result of misjudgments to the right of centre on these same tasks.


Extending the range of composite resin in everyday practice.
Composite resins, coupled with an acid-etch technique, can provide simple solutions to otherwise complex restorative problems. A catalogue of these possibilities is presented.


PI3K-Akt pathway-independent PIK3AP1 identified as a replication inhibitor of the African swine fever virus based on iTRAQ proteomic analysis.
African swine fever (ASF) is a severe infectious disease that has a high global prevalence. The fatality rate of pigs infected with ASF virus (ASFV) is close to 100%; in the absence of a safe and reliable commercial vaccine, this poses a threat to the global pig industry and public health. To better understand the interaction of ASFV with its host, isobaric tags for relative and absolute quantitation combined with liquid chromatography-mass spectrometry was used to conduct quantitative proteomic analysis of bone marrow-derived macrophage cells infected with ASFV. Overall, 4579 proteins were identified; 286 of these were significantly upregulated and 69 were significantly downregulated after ASFV infection. Gene Ontology, Kyoto Encyclopedia of Genes and Genomes, and protein-protein interaction network analyses were used to obtain insights into the dynamics and complexity of the ASFV-host interaction. In addition, immunoblotting revealed that the expression of PIK3AP1, RNF114, and FABP5 was upregulated and that of TRAM1 was downregulated after ASFV infection. Overexpression of PIK3AP1 and RNF114 significantly inhibited ASFV replication in vitro, but the suppressive effect of PIK3AP1 on ASFV replication was independent of the PI3K-Akt pathway. Further studies confirmed that ASFV MGF360-9L interacts with PIK3AP1 to reduce its protein expression level. Moreover, LY294002, an inhibitor of the PI3K-Akt pathway, inhibited ASFV replication, indicating the importance of the PI3K-Akt pathway in ASFV infection. This study identified the network of interactions between ASFV and host cells and provides a reference for the development of anti-ASFV strategies and for studying the potential mechanisms and pathogenesis of ASFV infection.


Proteomic Analysis Based on TMT Regarding the Therapeutic Action of Rhizoma Drynariae on Rats in an Osteoporosis Model.
Primary osteoporosis has increasingly become one of the risk factors affecting human health, and the clinical effect and action mechanism of traditional Chinese medicine in the treatment of primary osteoporosis have been widely studied. Previous studies have confirmed that in traditional Chinese medicine (TCM), Drynaria rhizome has a role in improving bone density. In this study, a tandem mass tag (TMT)-based proteomic analysis was conducted to derive potential targets for Drynaria rhizome treatment in postmenopausal osteoporosis. The model group (OVX) and experimental group (OVXDF) for menopausal osteoporosis were established using the universally acknowledged ovariectomy method, and the OVXDF group was given 0.48g/kg Rhizoma Drynariae solution by gavage for 12 weeks. After 12 weeks, femurs of rats selected for this study were examined with a bone mineral density (BMD) test, Micro-CT, ELISABiochemical testing, hematoxylin and eosin (HE) staining, and immunohistochemistry. A certain portion of the bone tissue was studied with a TMT-based proteomic analysis and functional and pathway enrichment analysis. Finally, key target genes were selected for Western blotting for validation. The comparison of the OVXDF and OVX groups indicated that Drynaria rhizome could improve bone density. In the TMT-based proteomic analysis, the comparison of these two groups revealed a total of 126 differentially expressed proteins (DEPs), of which 62 were upregulated and 64 were downregulated. Further, by comparing the differential genes between the OVXDF and OVX groups and between the OVX and SHAM groups, we concluded that the 27 differential genes were significantly changed in the rats selected for the osteoporosis model after Drynaria rhizome intragastric administration. The gene ontology (GO) enrichment analysis of DEPs showed that molecular function was mainly involved in biological processes, such as glucose metabolism, carbohydrate metabolism, immune responses, and aging. A Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of DEPs revealed that multiple differential genes were enriched in the estrogen and peroxisome proliferator-activated receptor (PPAR) signaling pathways. Relationships with nitrogen metabolism, glycerophospholipid metabolism, secretion systems, and tumor diseases were also observed. Western blotting was consistent with the analysis. We used TMT-based proteomics to analyze the positive effects of TCM Drynaria rhizome, which can regulate related proteins through the unique roles of multiple mechanisms, targets, and pathways. This treatment approach can regulate oxidative stress, improve lipid metabolism, reduce the inflammatory response mechanism, and improve bone density. These benefits highlight the unique advantages of TCM in the treatment of primary osteoporosis.


Sensitized phosphorescence as detection method for the enantioseparation of bupropion by capillary electrophoresis.
A new CE detection method was developed for the chiral drug bupropion (a second-generation antidepressant), based on phosphorescence both in the direct and in the sensitized mode using pulsed laser excitation at 266 nm. Electrokinetic chromatography using 5 mM sulfated-α-CD as chiral selector in 25 mM phosphate buffer at pH 3 allowed the separation of bupropion enantiomers with a high chiral resolution (Rs>3). In the sensitized phosphorescence detection mode, excitation energy is transferred from the analyte to an acceptor (1-bromo-4-napthhalenesulfonic acid or biacetyl) followed by time-resolved phosphorescence detection under deoxygenated buffer conditions. Using 2 × 10(-4) M biacetyl as the acceptor an LOD of 2 × 10(-7) M was obtained for each enantiomer, about 40 times better than in the direct mode. Under these separation conditions, no significantly different phosphorescence lifetimes (measured on-line) were obtained for the two bupropion enantiomers. The suitability of the method was demonstrated with the quantification of bupropion in a pharmaceutical formulation and its determination in a spiked urine sample.


Relevance of Laser Flare Photometry in Retinal Pathologies.
Laser flare photometry provides a non-invasive and objective measurement of the Tyndall effect in the anterior chamber. The laser flare value (measured in photon number per millisecond [pc/ms]) thus quantifies the extent of disruption to the blood-aqueous barrier and can be used in clinical applications to monitor uveitis therapy or to measure the postoperative degree of inflammation. Standardised performance must be observed during measurement. Publications of the last 35 years on laser flare photometry deal not only with the measurement procedure but also with its use in clinical practice for different ophthalmological pathologies. Likewise, various influencing factors have already been investigated and described that must be considered when measuring and evaluating laser flare values. The focus of this article is the relevance of laser flare photometry in retinal pathologies. In recently published studies, the level of objective tyndallometry in primary rhegmatogenous retinal detachment is shown to depend on lens status, symptom duration, and extent of retinal detachment. The greater is the area of the retina affected, the greater the disruption of the blood-aqueous barrier appears to be. Elevated laser flare values have also been considered as a predictor for the development of proliferative vitreoretinopathy (PVR). However, based on current knowledge, this assumption must be put into perspective. According to current data, objective tyndallometry can be used to monitor the progression of intraocular inflammation and to quantify the blood-aqueous barrier, and the values correlate with the extent and anatomical features, as well as the symptom duration in retinal detachment. Many influencing factors have already been identified. But further evaluation is desirable and needed. It is still unclear whether laser flare values can be used in the future as a predictor for sequelae such as PVR development.


PAH-induced metabolic changes related to inflammation in childhood asthma.
Epidemiological studies have shown that PAHs may exert adverse effects on childhood asthma. However, the underlying molecular mechanism remains to be fully elucidated. This study aimed to investigate this process in view of metabolic pathways, especially one-carbon metabolism and tryptophan metabolism. Fifty asthmatic children and 50 control subjects were recruited for this study. Serum IgE and IL-17A levels were detected by ELISA. Serum PAH concentrations were measured by GC-MS. One-carbon-related metabolites and tryptophan metabolites were determined by UPLC-Orbitrap-MS. DNA methylation was analyzed by bisulfite sequencing PCR. ChIP assays were used to examine H3K4me3 enrichment on IL-17A gene. Multivariable linear regression was performed to evaluate the association between PAHs and childhood asthma mediated by intermediators. HE staining in lung tissue, IgE and IL-17A in BALF, metabolic profiles in urine, and Ahr, Il-17a, and Cyp1a1 gene expression were determined in PAH-exposed mice. Serum Fla level was associated with childhood asthma (OR = 1.380, 95% CI: 1.063-1.792), and had a great effect on one-carbon metabolites, especially SAH, SAM, and Ser, which exerted significant mediation effects on the relationship between the Fla concentration and asthma. Moreover, we did find significant mediation effects between serum Fla and asthma by LINE-1 DNA methylation and H3K4me3 levels in the IL-17A promoter region. The differential Trp metabolites, such as Trp, tryptamine, IA, IAA, indole, IAld, and IAAld, indicated that asthmatic children had increased indole-AhR pathway. Mediation analysis failed to show a mediator effect of Trp metabolites in the association between PAHs and childhood asthma. An animal study confirmed that PAH exposure increased methylation levels, and altered Trp metabolite-AhR-IL-17A axis, which may be influenced by gender. PAHs disturbed one-carbon metabolism to influence the methyl group refilling DNA methylation and histone methylation, and disturbed tryptophan metabolism to regulate Th17-cell differentiation, which may elevate serum IL-17A concentration in asthmatic children.


Influence of left atrial function on Doppler transmitral and pulmonary venous flow patterns in dilated and hypertrophic cardiomyopathy: evaluation of left atrial appendage function by transesophageal echocardiography.
Information regarding the relation of left atrial (LA) function to transmitral and pulmonary venous (PV) flow is limited. Using transesophageal echocardiography, we analyzed this relation in 23 patients with dilated cardiomyopathy (DCM) and 25 patients with hypertrophic cardiomyopathy (HCM). Left atrial appendage (LAA) function was assessed as a substitute for overall LA function. Transmitral and PV flow-velocity variables, the LAA emptying flow velocity (LAA-A), and the LAA ejection fraction (LAA-EF) were determined. Each patient group was divided into two subgroups with a normal ( < 15 mm Hg) or elevated ( > or = 15 mm Hg) mean pulmonary wedge pressure (PWP). Transmitral and PV flow patterns as well as LA function were similar in the two subgroups with a normal PWP (11 patients with DCM and 14 patients with HCM). For the subgroups with an elevated PWP, however, the peak velocity ratio of the early filling wave (E) to atrial contraction wave (A) was higher in DCM patients (n = 12) than in HCM patients (n = 11) (2.1 +/- 0.7 vs 1.3 +/- 0.2; p < 0.01). This difference mostly resulted from a lower A velocity in the DCM group than in the HCM group (30 +/- 10 cm/sec vs 43 +/- 7 cm/sec; p < 0.05). In addition, the reverse flow velocity at atrial contraction in the PV was lower in the DCM group than in the HCM group (19 +/- 8 cm/sec vs 37 +/- 8 cm/sec; p < 0.01). These findings were associated with poorer LA systolic function in the DCM group (LAA-A, 35 +/- 13 cm/sec vs 60 +/- 11 cm/sec; LAA-EF, 37% +/- 12% vs 55% +/- 15%, p < 0.05, respectively). Our data suggest that a restrictive transmitral flow pattern develops more easily in DCM than in HCM because LA dysfunction is present in DCM, and that LA contractility plays an important role in determining the atrial contraction wave of transmitral and PV flows with elevated LA pressure.


Visio-Vestibular Deficits in Healthy Child and Adolescent Athletes.
To determine the relationship between patient characteristics and performance on the visio-vestibular examination (VVE) in a cohort of healthy youth athletes and explore the potential association between the VVE and other standardized concussion batteries. Cross-sectional. Suburban middle and high school. One hundred ninety subjects age 11 to 18 enrolled before their respective scholastic sport season between August 2017 and March 2020. Patient age, sex, concussion history, comorbidities, hours of weekly exercise, Sport Concussion Assessment Tool, 5th edition (SCAT-5), King-Devick (K-D), Postconcussion Symptom Inventory (PCSI). Visio-vestibular examination abnormalities (smooth pursuit, horizontal and vertical saccades, horizontal and vertical gaze stability, convergence, right and left monocular accommodation, complex tandem gait). Overall, 29.5% of subjects had at least one of 9 VVE elements abnormal, 7.9% at least 2, and 3.2% at least 3. None of 72 comparisons of the VVE elements, when stratified by age, sex, concussion history, history of headaches, attention deficit hyperactivity disorder, learning issues, psychiatric problems, motion sickness, or weekly hours of exercise, reached significance using the Benjamini-Hochberg procedure at a false discovery rate of 5%. There were no significant associations between VVE elements and the SCAT-5, K-D, or PCSI. The VVE is robust across multiple patient characteristics. Although healthy subjects may have one abnormal element, multiple abnormal elements are a less common feature, making multiple abnormal elements more indicative of concussion, highlighting the use of this assessment in the setting of injury. The VVE tests unique domains when compared with the PCSI, SCAT-5, and K-D.


Regional differences in dietary habits, coronary risk factors and mortality rates in Belgium. 1. Design and methodology. Nutrition and health: an interuniversity study.
The design and methodology are presented of a national survey that is in progress in Belgium relating dietary habits, coronary risk factors and mortality rates on a regional basis. The reasons and objectives of the study are explained. The sampling procedures and the survey methodology, in particular the measures of dietary habits, are presented and discussed.


Improved synaptic functionalities of Li-based nano-ionic synaptic transistor with ultralow conductance enabled by Al2O3barrier layer.
In this study, we investigated the effect of an Al2O3barrier layer in an all-solid-state inorganic Li-based nano-ionic synaptic transistor (LST) with Li3PO4electrolyte/WOxchannel structure. Near-ideal synaptic behavior in the ultralow conductance range (∼50 nS) was obtained by controlling the abrupt ion migration through the introduction of a sputter-deposited thin (∼3 nm) Al2O3interfacial layer. A trade-off relationship between the weight update linearity and on/off ratio with varying Al2O3layer thickness was also observed. To determine the origin of the Al2O3barrier layer effects, cyclic voltammetry analysis was conducted, and the optimal ionic diffusivity and mobility were found to be key parameters in achieving ideal synaptic behavior. Owing to the controlled ion migration, the retention characteristics were considerably improved by the Al2O3barrier. Finally, a highly improved pattern recognition accuracy (83.13%) was achieved using the LST with an Al2O3barrier of optimal thickness.


UK adaptive radiotherapy practices for head and neck cancer patients.
To provide evidence on the extent and manner in which adaptive practices have been employed in the UK and identify the main barriers for the clinical implementation of adaptive radiotherapy (ART) in head and neck (HN) cancer cases. In December 2019, a Supplementary Material 1, of 23 questions, was sent to all UK radiotherapy centres (67). This covered general information to current ART practices and perceived barriers to implementation. 31 centres responded (46%). 56% responding centres employed ART for between 10 and 20 patients/annum. 96% of respondents were using CBCT either alone or with other modalities for assessing "weight loss" and "shell gap," which were the main reasons for ART. Adaptation usually occurs at week three or four during the radiotherapy treatment. 25 responding centres used an online image-guided radiotherapy (IGRT) approach and 20 used an offline ad hoc ART approach, either with or without protocol level. Nearly 70% of respondents required 2 to 3 days to create an adaptive plan and 95% used 3-5 mm adaptive planning target volume margins. All centres performed pre-treatment QA. "Limited staff resources" and "lack of clinical relevance" were identified as the two main barriers for ART implementation. There is no consensus in adaptive practice for HN cancer patients across the UK. For those centres not employing ART, similar clinical implementation barriers were identified. An insight into contemporary UK practices of ART for HN cancer patients indicating national guidance for ART implementation for HN cancer patients may be required.


Effect of surfactants on casein structure: a spectroscopic study.
Fluorescence and circular dichroism spectroscopy were used to study the effect of two surfactants having oppositely charged head groups - cationic cetyltrimethyl ammonium bromide (CTAB) and anionic sodium dodecyl sulphate (SDS) - on the structure of the intrinsically unstructured proteins alpha s-, beta- and kappa-caseins. Although globular proteins are generally known to denature on interacting with surfactants, the caseins were found to adopt more ordered conformations in presence of both SDS and CTAB. The folding induced by CTAB was more efficient than by SDS, as implied by the behaviour of fluorescence and circular dichroic spectra of the caseins in solutions containing varying concentrations of the surfactants. The differential response of the proteins to the two surfactants may lie in the fact that the negatively charged caseins experience a repulsive electrostatic interaction with the negatively charged head groups of SDS, while their interaction with the positively charged head groups of CTAB is attractive in nature. Our results are consistent with two different types of the 'necklace and bead' model for the structure of surfactant-casein complexes: while groups of SDS molecules converge tail first around exposed hydrophobic surfaces of the caseins to form micelle-like structures, the protein itself wraps around micellar aggregates of CTAB that have cationic head groups in close association with its negatively charged/polar residues.


Matriptase does not require hepatocyte growth factor activator inhibitor type-1 for activation in an epithelial cell expression model.
Matriptase is a type II transmembrane serine protease. Paradoxically, activation of this protease is thought to require its physiological inhibitor, hepatocyte growth factor activator inhibitor type-1 (HAI-1). In the present study, however, we obtained evidence in a stable transfection experiment using Madin-Darby canine kidney cells that matriptase activation does not require HAI-1.


A new technique for ureteral stent placement during pregnancy using endoluminal ultrasound.
Endoluminal sonography using the IVUS is a minimally invasive procedure performed using a flexible 6.2F catheter with a 20 megahertz transducer that can easily be passed endoscopically to image the urethra, bladder, ureter and renal pelvis. In the current study, a woman patient in her third trimester of pregnancy with a renal pelvic stone and persistent colic was safely treated with placement of an indwelling ureteral stent. Because it is minimally invasive and does not require the use of roentgenograms, we believe the IVUS will be a useful tool to assist with the management of ureteral colic in patients who are pregnant in whom intervention is required.


Atrial-selective drugs for treatment of atrial fibrillation.
Atrial fibrillation (AF) is accompanied by a high risk of thromboembolic complications necessitating anticoagulation therapy. Arrhythmias have a high tendency to become persistent. Catheter ablation techniques are highly effective in the treatment of AF; however, these procedures are far too costly and time-consuming for the routine treatment of large numbers of AF patients. Moreover, many patients prefer drug treatment although conventional antiarrhythmic drugs are moderately effective and are burdened with severe cardiac and noncardiac side effects. New antifibrillatory drugs developed for the treatment of AF include multichannel blockers with a high degree of atrial selectivity. The rationale of this approach is to induce antiarrhythmic actions only in the atria without conferring proarrhythmic effects in the ventricles.Atrial selective drug action is expected with ion channel blockers targeting ion channels that are expressed predominantly in the atria, i.e., Kv1.5 (I(Kur)), or Kir 3.1 and Kir 3.4 (I(K,ACh)). Na(+) channel blockers that dissociate rapidly may exert atrial selectivity because of subtle differences in atrial and ventricular action potentials. Finally, atrial-selective targets may evolve due to disease-specific processes (e.g., rate-dependent Na(+) channel blockers, selective drugs against constitutively active I(K,ACh) channels).


Tectal induction of cortical arousal: evidence implicating multiple output pathways.
The rodent superior colliculus mediates a wide range of physiological and behavioural responses to sudden stimuli, including desynchronisation of the cortical electroencephalogram (EEG). To investigate how this desynchronisation is produced, one of two powerful excitatory agents, sodium L-glutamate (200 nl, 10 nmol) or bicuculline methiodide (200 nl, 40 pmol), was injected into the dorsal midbrain of sleeping rats. Microinjections at sites widely distributed throughout all layers of the superior colliculus were able to desynchronise the cortical EEG. i) In the superficial layers, bicuculline was effective at more sites than glutamate, whereas the reverse was true for the deep layers. ii) At some sites EEG desynchronisation occurred together with the defensive or orienting movements that are obtained from collicular stimulation in awake animals. At other sites cortical arousal occurred without such movements. iii) Comparison with a previous study suggested that urethane selectively blocks cortical arousal to glutamate injections in the superficial and intermediate grey layers. This evidence suggests that multiple collicular output pathways can desynchronise the cortical EEG, perhaps reflecting multiple functions for EEG desynchronisation.


The experience and expression of anger in posttraumatic stress disorder: the relationship with metacognition.
Anger experience and expression are a common issue in those experiencing PTSD. However, it remains unclear what variables affect anger and its expression in PTSD. To explore the relationships of synthetic forms of metacognition and metacognitive beliefs with anger experience and expression in PTSD, independent of the effects hyperarousal and depression symptoms. Participants were 51 veterans with diagnosed with PTSD. Metacognition was assessed using the Metacognition Assessment Scale-Abbreviated (MAS-A) and the Metacognitions Questionnaire (MCQ). Depression, PTSD symptom severity, and seven domains of anger expression were also assessed. Correlations showed after controlling for overall levels of hyperarousal, higher MAS-A total scores were related to lower levels of State Anger, Feeling Angry, Expressing Anger Physically, and Anger Expression in. Lower MCQ scores were related to lower State anger, Expressing anger verbally, and Expressing anger physically. Higher levels of depression were related to higher levels of Trait anger, Expressing anger physically, Anger expression out, and Anger expression in. Multiple regressions suggested that the MAS-A and MCQ predicted unique portions of the variance in anger experience and expression. Metacognitive deficits may affect anger experience and expression in those with PTSD and may be an important treatment target.


Regulating Hetero-Nucleation Enabling Over 14% Efficient Kesterite Solar Cells.
Developing well-crystallized light-absorbing layers remains a formidable challenge in the progression of kesterite Cu2 ZnSn(S,Se)4 (CZTSSe) solar cells. A critical aspect of optimizing CZTSSe lies in accurately governing the high-temperature selenization reaction. This process is intricate and demanding, with underlying mechanisms requiring further comprehension. This study introduces a precursor microstructure-guided hetero-nucleation regulation strategy for high-quality CZTSSe absorbers and well-performing solar cells. The alcoholysis of 2-methoxyethanol (MOE) and the generation of high gas-producing micelles by adding hydrogen chloride (HCl) as a proton additive into the precursor solution are successfully suppressed. This tailored modification of solution components reduces the emission of volatiles during baking, yielding a compact and dense precursor microstructure. The reduced-roughness surface nurtures the formation of larger CZTSSe nuclei, accelerating the ensuing Ostwald ripening process. Ultimately, CZTSSe absorbers with enhanced crystallinity and diminished defects are fabricated, attaining an impressive 14.01% active-area power conversion efficiency. The findings elucidate the influence of precursor microstructure on the selenization reaction process, paving a route for fabricating high-quality kesterite CZTSSe films and high-efficiency solar cells.


Would blockage of cytokines improve the outcome of pancreatic islet transplantation?
It has been estimated that up to 60% of pancreatic islet tissue undergoes apoptosis within the first several days post-transplantation. This strongly suggests the involvement of an inflammatory event other than alloantigen-specific immune reaction following islet transplantation which contributes to partial destruction of grafts. Inflammatory cytokines including IL-1beta, TNF-alpha and IFN-gamma are implicated in the pancreatic islet beta-cell death and functional loss during autoimmune diabetes and also seem to be involved in early loss of islet mass in islet transplantation. Inflammatory cytokines and free oxygen radicals released in situ could cause apoptosis and the functional impairment of islets after islet transplantation and graft failure. It can be hypothesized that preventing destruction of transplanted islets using cytokine blockade could be helpful in improving islet transplantation outcome. Several approaches have been made based on this hypothesis to examine the effect of inflammatory blockade on the islets survival and functional islet mass. Further investigations are required to identify most efficient way for block of cytokine-induced damage in pancreatic islets transplantation.


Integrated carbon fiber electrodes within hollow polymer microneedles for transdermal electrochemical sensing.
In this study, carbon fiber electrodes were incorporated within a hollow microneedle array, which was fabricated using a digital micromirror device-based stereolithography instrument. Cell proliferation on the acrylate-based polymer used in microneedle fabrication was examined with human dermal fibroblasts and neonatal human epidermal keratinocytes. Studies involving full-thickness cadaveric porcine skin and trypan blue dye demonstrated that the hollow microneedles remained intact after puncturing the outermost layer of cadaveric porcine skin. The carbon fibers underwent chemical modification in order to enable detection of hydrogen peroxide and ascorbic acid; electrochemical measurements were demonstrated using integrated electrode-hollow microneedle devices.


TABADO 2 : une stratégie d'accompagnement au sevrage tabagique des adolescents en milieu scolaire.
La consommation de tabac est une des premières causes de mortalité évitable au niveau mondial. La dépendance au tabac s'observe dès l'adolescence. En complément des actions visant à prévenir l'entrée des jeunes dans le tabagisme, il est donc nécessaire de développer des stratégies spécifiques pour les adolescents déjà fumeurs. Un programme d'accompagnement au sevrage tabagique pour adolescents, TABADO, a montré son efficacité lors d'un essai contrôlé mené dans des centres de formation des apprentis en 2007/2009. En 2018, l'Institut national du cancer a souhaité généraliser TABADO au niveau national et l'étendre aux lycées professionnels. Pour accompagner ce processus de mise à l'échelle, il était essentiel d'élaborer la théorie d'intervention et d'analyser la transférabilité de TABADO en conditions de vie réelle et dans de nouveaux contextes, et de proposer les adaptations correspondantes. Une recherche spécifique a été conduite et détaillée par ailleurs. L'objectif de cette publication pragmatique est de présenter la nouvelle stratégie TABADO 2 aux acteurs et décideurs de santé publique, et le guide afférent développé pour les accompagner dans la mise en œuvre.Une étude de cas multiples (n = 10) a été menée à partir de la mise en œuvre de TABADO dans trois régions françaises, qui reposait sur des observations, entretiens et séminaires de retour d'expérience. Elle a mis en évidence l'adaptation de l'intervention aux contextes locaux, ainsi que de nouveaux leviers interventionnels mis en œuvre. De plus, les investigations ont montré que pour instaurer un climat favorable à la démarche d'arrêt de la consommation de tabac et soutenir son maintien sur la durée, il était nécessaire d'inscrire TABADO dans une stratégie globale de l'établissement scolaire et de son environnement. Cette démarche a permis de proposer une transformation de l'intervention TABADO en une nouvelle stratégie - TABADO 2 - et de proposer un guide pour accompagner son déploiement national.


ACCURACY OF INTRAORAL SCANNERS VERSUS TRADITIONAL IMPRESSIONS: A RAPID UMBRELLA REVIEW.
This study aimed to (1) report the trueness and precision of intraoral scanning (IOS) in dentistry based on recent secondary sources and to (2) appraise the reporting quality of the titles and abstracts of the included literature. This rapid overview searched the PubMed/Medline and Cochrane Database of Systematic Reviews in March 2021 to identify reviews reporting on the accuracy of IOS. The reference list from the eligible studies was also screened for identification of other potentially eligible studies. The inclusion criteria consisted of English language systematic reviews or meta-analyses published between 2019 and 2021. The exclusion criteria were primary studies, narrative review, and extraoral scanners. The assessment of reporting quality of abstracts of systematic reviews was performed using the reporting checklist PRISMA extension for Abstracts (PRISMA-A). This was a self-funded research project. Out of the full text screened 25 records, 11 reviews were included. Most studies supported the IOS approach being as precise and accurate as the conventional one. Only one study significantly favored the conventional approach over the IOS, and two studies abstained from making a recommendation. The IOS was significantly superior to the traditional technique in terms of patient preference and time efficiency. After applying PRISMA-A, recommendations for improvements on titles and abstracts of future reviews of IOS and conventional impressions are provided. Laboratory data indicated similar accuracy between IOS and conventional impressions, whereas clinical data found the same in less than 4-unit fixed dental prostheses. For more extensive definitive fixed solutions or removable prostheses, the conventional approach is recommended. IOS was superior in terms of patient preference and time reduction. More clinical trials are required to determine the clinical effectiveness of incorporating IOS in broader scenarios. Better quality of reporting secondary sources abstract is advised.


Salvage regional therapy using hepatic artery infusion pump in unresectable chemotherapy resistant colorectal liver metastases.
Little is known about the influence of hepatic artery infusion pump (HAIP) therapy in the setting of chemotherapy resistant hepatic disease in the era of modern systemic therapies. Patients who underwent HAIP therapy for chemotherapy resistant and unresectable colorectal liver metastases (CRLM) were reviewed retrospectively. A total of 25 patients met inclusion criteria. 52% had isolated CRLM and 92% had five or more metastatic lesions. Partial response was noted in 40% of patients. Median hepatic progression-free survival (PFS) was 7 months in those with extrahepatic disease versus 6 months in those with isolated CRLM at the time of HAIP placement (p = 0.75). Median overall survival was 8 months in patients with extrahepatic disease and 14 months in patients with isolated CRLM (p = 0.06). Our findings are comparable to published data and augment the literature which supports HAIP use in chemotherapy-resistant, liver-predominant metastatic colorectal cancer patients.


Purification and properties of nitrogenase in ethylene glycol at sub-zero temperatures.
Both the protein components Kp1 and Kp2 of nitrogenase from Klebsiella pneumoniae were found to be stable in aq. 50% (v/v) ethylene glycol at +30 degrees C or below. At -20 degrees C in this medium their sensitivities to O2 were diminished somewhat. Though purification could be carried out at -20 degrees C, the product had the same specific activity and was obtained in the same yield as when the purification was carried out by standard procedures. This suggests that such procedures yield enzyme undamaged in the course of the purification by O2, thermal denaturation or proteolytic digestion.


Detection of blood coagulation and clot formation using quantitative ultrasonic parameters.
The ultrasonic parameters of sound velocity, attenuation and integrated backscatter were applied to detect the process of coagulation and clot formation in porcine blood. Fresh porcine blood containing 15% anticoagulant solution was collected. Blood samples with a hematocrit of 45% were obtained by reconstituting the packed erythrocytes with the separated plasma for ultrasound measurements performed with a 10-MHz focused transducer. A 24-mL aliquot of blood was placed in a container and 12 mL of 0.2 mol/L CaCl2 solution was added to induce clot formation. In each measurement, radio-frequency signals of the blood digitized at 100 MHz were collected for 50 min at a temporal resolution of 1 A-line per s. Results showed that all of the parameters increased within the initial 3 min and, then, immediately decreased dramatically as the CaCl2 solution was added. Subsequently, the sound velocity gradually increased with time and the integrated backscatter and attenuation increased in accordance with blood coagulation until approximately 500 and 2600 s, respectively. The integrated backscatter, attenuation and sound velocity can be divided into different stages, including red cell aggregation, reduction in hematocrit, blood coagulation and clot formation, corresponding to variations in the physical and chemical properties of the blood. The integrated backscatter, attenuation and sound velocity increased because of the changes in blood properties during the process of coagulation and clot formation: by 8.2 dB, 0.65 dB/cm, and 0.6%, respectively. These results provide a feasibility for further applying ultrasonic parameters to in vivo monitor the progress of clotting and thrombosis research.


Depression and Anxiety in Preschoolers: A Review of the Past 7 Years.
This article reviews recent empirical literature on the prevalence, correlates, assessment, and treatment of preschool-onset internalizing disorders. Major advances in the acceptance and recognition of both preschool-onset depression and anxiety have occurred over the past decade. This work has been greatly enhanced by the discovery of genetic, neural, and physiologic indicators, which further validate these constellations of symptoms in young children. Despite this growth in research, much work still needs to be done to further elucidate the cause, risk, treatment, and protective factors for preschool-onset internalizing disorders.


A comparison of emergency triage scales in triaging poisoned patients.
Triage of toxicology patients presents a challenge due to their complexity, underlying psychosocial issues, and additional pharmacological considerations. Two emergency department triage systems used in Australia, the Australasian Triage Scale (ATS) and the Manchester Triage System (MTS), were compared in triaging patients presenting with poisoning and envenoming. In this simulation-based study, 30 triage nurses from three hospitals were given 8 tabletop scenarios and asked to provide a triage category. 20 nurses from two hospitals using the ATS, and 10 nurses from a third hospital using the MTS, triaged 8 scenarios, grouped into "commonly encountered" (n=4) and "rarely encountered" (n=4). Triage systems and scenario groups were compared for median triage category and variance in scoring. Triage nurses also noted if they would seek help from toxicology services or the poisons information centre (PIC) for advice. Overall, MTS nurses triaged all 8 scenarios with a lower acuity triage category, though statistically significant for only 3 scenarios. ATS nurses scored higher acuity triage category in all 4 "rare" highly toxic presentations, whereas MTS nurses scored higher acuity when vital signs were abnormal. MTS showed wider variance in triage scores in both scenario groups when compared to the ATS. Triage nurses without access to local toxicology services chose to contact PIC in most cases. When compared to the ATS, MTS gave a lower acuity triage score for all common and rarely encountered poisoning scenario groups, which included highly toxic ingestions that appear well at triage but may progress to severe poisoning. Triage nurses should refer to information on highly toxic exposures and envenomation guidelines during their triage risk assessment.


Comparison of Mortality Rate and Severity of Pulmonary Involvement in Coronavirus Disease-2019 Adult Patients With and Without Type 2 Diabetes: A Cohort Study.
Patients with diabetes are potentially at higher risk of mortality due to coronavirus disease-2019 (COVID-19). In this study, we aimed to compare the outcomes and severity of pulmonary involvement in COVID-19 patients with and without diabetes. In this cohort study, we recruited patients with diabetes who were hospitalized due to COVID-19 during the period from February 2020 to May 2020. Hospitalized individuals without diabetes were enrolled as control subjects. All patients were followed for 90 days and clinical findings and patients' outcomes were reported. Over a period of 4 months, 127 patients with diabetes and 127 individuals without diabetes with a diagnosis of COVID-19 were recruited. Their mean age was 65.70±12.51 years. Mortality was higher in the group with diabetes (22.8% vs 15.0%; p=0.109), although not significantly. More severe pulmonary involvement (p=0.015), extended hospital stay (p<0.001) and greater need for invasive ventilation (p=0.029) were reported in this population. Stepwise logistic regression revealed that diabetes was not independently associated with mortality (p=0.092). Older age (odds ratio [OR], 1.054; p=0.003), aggravated pulmonary involvement on admission (OR, 1.149; p=0.001), presence of comorbidities (OR, 1.290; p=0.020) and hypothyroidism (OR, 6.576; p=0.021) were associated with mortality. Diabetic foot infection had a strong positive correlation with mortality (OR, 49.819; p=0.016), whereas insulin therapy had a negative correlation (OR, 0.242; p=0.045). The mortality rate due to COVID-19 did not differ significantly between patients with or without diabetes. Older age, macrovascular complications and presence of comorbidities could increase mortality in people with diabetes. Insulin therapy during hospitalization could attenuate the detrimental effects of hyperglycemia and improve prognosis of patients with COVID-19 and diabetes.


Kidney involvement in MELAS syndrome: Description of 2 cases.
MELAS syndrome -myopathy, encephalopathy, lactic acidosis and stroke-like episodes- is a maternally-inherited mitochondrial cytopathy related to several mitochondrial DNA mutations, with the A3243G mutation in tRNALeu gene being the most frequent of them. Apart from its typical symptomatology, patients usually exhibit a maternally-inherited history of neurosensory deafness and insulin-dependent type 2 diabetes mellitus (T2DM). Recent studies have shown that few patients carrying a A3243G mutation also suffer from renal dysfunction, usually in form of focal segmental glomerulosclerosis (FSGS). In this study we examine kidney involvement in 2 unrelated patients with a A3243G mutation by genetic testing. Both have a maternally-inherited neurosensory deafness and insulin-dependent T2DM. A renal biopsy was performed in both patients. One patient developed nephrotic proteinuria and renal insufficiency, with FSGS findings being observed in the kidney biopsy, whereas the other suffered from mild proteinuria and renal insufficiency, with non-specific glomerular changes. The presence of FSGS or other kidney involvement accompanied by hereditary neurosensory deafness and T2DM could be suggestive of a A3243G tRNALeu mutation and should prompt a genetic testing and an evaluation of potential extrarenal involvement.


Multiple antibiotic resistance (MAR) index and its reversion in Pseudomonas aeruginosa.
The MAR indexes of hospital isolates of Pseudomonas aeruginosa were determined with reference to nine different cephalosporins. The values for all the strains were higher than 0.2 suggesting their origin from a high risk source of contamination where antibiotics are often used. Emergence of MAR pathogenic strains of Ps. aeruginosa indicated possible nosocomial infection in the hospital environment. beta-Lactamases produced by these organisms were tested and their inhibition by clavulanic acid was studied. beta-Lactamase produced by one of these strains (Ps-1) could not be inhibited by clavulanic acid whereas beta-lactamases of three other strains (Ps-2, Ps-3 and Ps-4) could be inhibited by clavulanic acid in the presence of cephalosporins, suggesting a possible use of clavulanic acid in combination with cephalosporins, to combat beta-lactamase induced resistance in Ps. aeruginosa.


Bioavailability of intramuscular rocuronium in infants and children.
Intramuscular rocuronium, in doses of 1,000 microg/kg in infants and 1,800 microg/kg in children, produces complete twitch depression in 5-6 min. To determine the rate and extent of absorption of rocuronium after intramuscular administration, blood was sampled at various intervals after rocuronium administration by both intramuscular and intravenous routes to determine plasma concentrations (Cp) of rocuronium. Twenty-nine pediatric patients ages 3 months to 5 yr were anesthetized with N2O and halothane. The trachea was intubated, ventilation was controlled, and adductor pollicis twitch tension was measured. When anesthetic conditions were stable, rocuronium (1,000 microg/kg for infants and 1,800 microg/kg for children) was injected either intramuscularly (in the deltoid muscle) or intravenously. Four venous plasma samples were obtained from each child 2-240 min after rocuronium administration. A mixed-effects population pharmacokinetic analysis was applied to these values to determine bioavailability, absorption rate constant, and time to peak Cp with intramuscular administration. With intramuscular administration, rocuronium's bioavailability averaged 82.6% and its absorption rate constant was 0.105 min(-1). Simulation indicated that Cp peaked 13 min after rocuronium was given intramuscularly, and that 30 min after intramuscular administration, less than 4% of the administered dose remained to be absorbed from the intramuscular depot. After rocuronium is administered into the deltoid muscle, plasma concentrations peak at 13 min, and approximately 80% of the administered drug is absorbed systemically.


Complete PacBio Single-Molecule Real-Time Sequence of a Novel Probiotic-Like Bacterium, Rouxiella badensis subsp. acadiensis, Isolated from the Biota of Wild Blueberries in the Acadian Forest.
The PacBio whole-genome sequencing of the potential probiotic strain Canan SV-53T recovered from lowbush blueberries demonstrates high homology to Rouxiella badensis but with distinct enough clusters to propose the name Rouxiella badensis subsp. acadiensis. The sequencing also detected the presence of two native plasmids.


Antigenic Variation of Avian Influenza A(H5N6) Viruses, Guangdong Province, China, 2014-2018.
Market surveillance showed continuing circulation of avian influenza A(H5N6) virus in live poultry markets in Guangdong Province in 2017, despite compulsory vaccination for avian influenza A(H5Nx) and A(H7N9). We analyzed H5N6 viruses from 2014-2018 from Guangdong Province, revealing antigenic drift and decreased antibody response against the vaccine strain in vaccinated chickens.


Endothelin A receptor antagonist LU 135252 inhibits hypercholesterolemia-induced, but not deendothelialization-induced, atherosclerosis in rabbit arteries.
The purpose of the study was to test the capability of the endothelin A receptor antagonist LU 135252 to reduce neointimal formation in rabbits after balloon denudation with and without the presence of hypercholesterolemia. Twenty-eight male New Zealand White rabbits underwent balloon denudation of the infrarenal aorta. The animals were randomly assigned to 1 of the 4 groups. After balloon denudation, group 1 (n = 6) and 2 (n = 7) received a standard diet, and group 3 (n = 8) and 4 (n = 7) were fed a 0.5% cholesterol diet. All interventional procedures were performed while the rabbits were under general anesthesia. One week prior to intervention treatment with LU 135252 was started in group 2 and 4. After 6 weeks the animals were killed for morphometric and histological analysis. Rabbits in all treatment groups developed neointimal hyperplasia. By additional systemic treatment with LU 135252, the mean neointima to media ratio was significantly reduced only in the hypercholesterolemic animals of group 4 (neointimal to media ratio area of group 3 vs group 4: 2.07 +/- 0.62 vs 1.41 +/- 0.45, P < 0.05). ET receptor blockade in group 2 and 4 did not have an effect on plasma levels of cholesterol, very low-density lipoprotein-, high-density lipoprotein-, and low-density lipoprotein-cholesterol. LU 135252 was efficient in reducing lipid induced atherosclerotic changes but was ineffective in inhibiting restenosis induced by balloon denudation.


Pixantrone (Novuspharma).
Novuspharma is developing pixantrone, a second-generation, well-tolerated anthracenedione analog, for the potential treatment of lymphoma and multiple sclerosis.


Transition to adult rheumatology care is necessary to maintain DMARD therapy in young people with juvenile idiopathic arthritis.
To evaluate the outpatient health service utilisation of young people (YP) with juvenile idiopathic arthritis (JIA) during transfer from paediatric to adult care in Germany. Based on claims data of a large German statutory health fund (BARMER GEK) we conducted a cohort study of 2008-2014. Insured YP with diagnosis of JIA (ICD: M08 or M09.0) in two quarters at the age of 16 and continuous insurance until the age of 20 were included. Outpatient health care utilisation, prescribed antirheumatic drugs and diagnosis maintenance were analysed over four consecutive years for all YP with JIA and for the subgroup on disease-modifying antirheumatic drugs (DMARDs) at the age of 16. A total of 256 YP with JIA (67% female) were followed during the transfer period, 115 of them received (DMARDs) at the age of 16. At the age of 20, 30% of all 256 YP utilised adult specialty care, 59% still had a JIA diagnosis and 30% were treated with DMARDs. Of those 115 YP, treated with DMARDs at the age of 16 and in need of follow-up, 49% had visited an adult rheumatologist at the age of 20, 75% had a JIA diagnosis and 56% were on DMARDs. Those 49% YP with successful transfer to adult specialty care had a JIA diagnosis in 93% and were treated with DMARDs in 80%. YP, who were only seen by general practitioners, had a JIA diagnosis in 54% and received DMARDs in 27%. One in two YP with JIA and a clear need for ongoing follow-up do not utilise adult rheumatologic care. The maintenance of JIA diagnosis and DMARD therapy depend on the utilisation of specialised care services.


Association between high collagenase-3 expression levels and poor prognosis in patients with head and neck cancer.
Squamous cell carcinoma of the head and neck (HNSCC) is a common cancer type. The ability for curative treatment with surgery and radiotherapy (RT) is usually highly dependent on tumor stage at the time of diagnosis. The purpose of this study was to determine whether the expression of a cancer-specific proteinase, collagenase-3 (matrix metalloproteinase-13 [MMP-13]), is associated with survival parameters in patients with HNSCC. We studied MMP-13 expression in tumors of 81 patients with stage I-IV HNSCC treated with surgery alone or in combination with radiotherapy. We found a subgroup of patients with high MMP-13 expression level in their tumors (>/=90% MMP-13-positive tumor cells) associated with unfavorable prognosis (median overall survival [OS], 11.8 vs 19.6 months, p = .032). In addition, the median disease-specific survival (DSS) time was markedly reduced in this subgroup (13.8 months vs 40.7 months, p = .062). When the subgroup of patients treated with a curative intent was studied, the same association was found in OS (13.8 vs 24.6 months, p = .023) and DSS (p = .004). In addition, there was a trend for association between >/=90% MMP-13 positivity and a recurrent tumor (p = .078) in curatively treated patients. The short survival time associated with high MMP-13 expression levels could not be predicted by tumor size or local lymph node invasion. These results show that a high MMP-13 expression level is associated with aggressiveness of HNSCC and may have prognostic value in patient evaluation.


Genotype-phenotype correlation of 17 cases of Pompe disease in Spanish patients and identification of 4 novel GAA variants.
Pompe disease (PD) is an autosomal recessive metabolic disorder caused by pathogenic variants in the acid α-glucosidase gene (GAA) that produces defects in the lysosomal acid α-1,4-glucosidase. We aimed to identify genetic variations and clinical features in Spanish subjects to establish genotype-phenotype correlation. A total of 2637 samples of patients who showed symptoms or susceptible signs of PD were enrolled in this observational study. Enzymatic activity was detected by fluorometric techniques and the genetic study was carried out using Next-Generation Sequencing. Fourteen different variants from 17 diagnosed patients were identified, seven males and nine females with LOPD (mean age 36.07, SD 20.57, range 7-64) and a 2-day-old boy with IOPD, four genetic variants had not been described in the literature previously, including a homozygous variant. In all of them α-glucosidase activity was decreased. Muscle weakness, respiratory distress, exercise intolerance, hypotonia, dysphagia and myalgia were commonly observed in patients. This study report four new genetic variants that contribute to the pathogenic variants spectrum of the GAA gene. We confirm that patients in Spain have a characteristic profile of a European population, with c.-32-13T>G being the most prevalent variant. Furthermore, it was confirmed that the c.236_246delCCACACAGTGC pathogenic variant in homozygosity is associated with early disease and a worse prognosis.


Angioembolization of Post-traumatic Intrahepatic Arterioportal Fistula Presenting With Portal Hypertension.
Traumatic hepatic arterioportal fistula is an abnormal communication between the hepatic artery and portal vein and is a rare cause of non-cirrhotic portal hypertension with delayed presentation, usually after a remote history of abdominal trauma or an interventional procedure. This case report is of one such rare presentation, wherein a 59-year-old gentleman presented with unexplained ascites and complications of portal hypertension, eventually diagnosed with an arterioportal fistula on a computed tomography scan and managed by angioembolization. There was a remarkable improvement in the complications of portal hypertension after the coil embolization.


Hemodynamic assessment of oral peripheral vasodilator therapy in chronic congestive heart failure: prolonged effectiveness of isosorbide dinitrate.
To evaluate the effectiveness of oral vasodilator therapy in chronic congestive heart failure, 20 mg of isosorbide dinitrate or placebo was administered orally in double-blind fashion to 25 patients with congestive heart failure. In 15 patients receiving isosorbide dinitrate, pulmonary arterial wedge pressure decreased 5 minutes to 5 hours after drug administration; the peak reduction was observed at 1 hour (from 23 to 14 mm Hg; P less than 0.001). Wedge pressure decreased to normal (12 mm Hg or less) in 8 of the 15 patients (Group I) but remained greater than 12 mm Hg in 7 (Group II). Reductions in mean systemic arterial pressure, systemic vascular resistance and pressure-time per minute also occurred. Indexes of pump output were unchanged in the 15 who received isosorbide dinitrate but tended to decrease slightly in Group I. Stroke index (from 23 to 26 cc/m2) and stroke work index (from 21.4 to 24.1 g-m/m2) increased slightly but significantly (P less than 0.05) in Group II. Thus the prinicpal hemodynamic action of isorbide dinitrate is marked and sustained reduction in left ventricular filling pressure without pronounced effect on cardiac output. This agent should be used in congestive heart failure primarily for relief of congestive symptoms.


Regioselective synthesis of cytarabine monopropionate by using a fungal whole-cell biocatalyst in nonaqueous medium.
The utilization of a dehydrated fungal biocatalyst of Aspergillus oryzae cells was successfully performed to achieve efficient acylation modification of a polar nucleoside cytarabine (ara-C). Organic solvents showed evident influence on the reaction catalyzed by the A. oryzae whole-cells. Except for hexane-pyridine, the catalytic activity and regioselectivity of the whole-cells clearly increased with increasing the polarity of the hydrophobic organic solvents used. The effects of some crucial factors on the reaction were further examined. The best reaction medium, hydrophobic solvent concentration, vinyl propionate/ara-C ratio, reaction temperature and shaking speed were confirmed as isopropyl ether (IPE)-pyridine, 30% (v/v), 90, 30 °C and 140-180 rpm, respectively. The cell biocatalyst also showed good thermal stabilities in both IPE-pyridine and hexane-pyridine systems. In addition, the desired 3'-O-propional derivative of ara-C was synthesized with the yields of 88.3% and regioselectivity (>70%). The resulting biocatalytic system appears to be an effective alternative, and can thus be employed for application in highly regioselective modification of nucleoside analogues.


Uncertainty in the perioperative management of high-risk cutaneous squamous cell carcinoma among Mohs surgeons.
To evaluate whether Mohs surgeons' management of high-risk cutaneous squamous cell carcinoma (HRCSCC) is uniform regarding radiologic nodal staging (RNS) and adjuvant radiation therapy (ART). A survey study of randomly selected, fellowship-trained Mohs surgeons. An academic medical center. American College of Mohs Surgery members who responded to an e-mail invitation completed either a survey regarding management of HRCSCC (n=117) or SCC with perineural invasion (PNI) (n=118). Participants totaled approximately 25% of the American College of Mohs Surgery membership. (1) Percentage of patients with HRCSCC referred for RNS, sentinel lymph node biopsy (SLNB), or ART over the preceding 12 months; (2) top factors leading surgeons to consider RNS, SLNB, or ART; and (3) acceptance of ART for clinical scenarios of various degrees of PNI. Most respondents cited PNI and in-transit metastasis as top factors leading to consideration of RNS, SLNB, or ART. Otherwise, there was no consensus regarding use of, or indications for, RNS, SLNB, or ART. The lack of consistency between experts indicates that there is equipoise regarding indications for RNS and ART in HRCSCC. There is also wide variation in RNS and ART practices among Mohs surgeons who are specifically trained to manage HRCSCC. Clinical trials should therefore be conducted in these areas as there is no clear standard of care.


Dynamical analysis of the fission yeast cell cycle via Markov chain.
The cell cycle is a complex network involved in the regulation of cell growth and proliferation. Intrinsic molecular noise in gene expression in the cell cycle network can generate fluctuations in protein concentration. How the cell cycle network maintains its robust transitions between cell cycle phases in the presence of these fluctuations remains unclear. To understand the complex and robust behavior of the cell cycle system in the presence of intrinsic noise, we developed a Markov model for the fission yeast cell cycle system. We quantified the effect of noise on gene and protein activity and on the probability of transition between different phases of the cell cycle. Our analysis shows how network perturbations decide the fate of the cell. Our model predicts that the cell cycle pathway (subsequent transitions from [Formula: see text]) is the most robust and probable pathway among all possible trajectories in the cell cycle network. We performed a sensitivity analysis to find correlations between protein interaction weights and transition probabilities between cell cycle phases. The sensitivity analysis predicts how network perturbations affect the transition probability between different cell cycle phases and, consequently, affect different cell fates, thus, forming testable in vitro/in vivo hypotheses. Our simulation results agree with published experimental findings and reveal how noise in the cell cycle regulatory network can affect cell cycle progression.


Structure-activity study of novel tricyclic benzazepine arginine vasopressin antagonists.
Novel tricyclic benzazepine derivatives were synthesized as arginine vasopressin (AVP) antagonists. Several tricyclic compounds showed potent antagonistic activity in rat AVP receptors V(1a) and V(2). Derivatives containing pyrrolo-tricyclic amines, 13i-k, 30, and 31 also showed selectivity for the V(2) receptor.


Optimizing Door-to-Groin Puncture Time: The Mayo Clinic Experience.
To provide a better understanding of methods that can be used to improve patient outcomes by reducing the door-to-groin puncture (DTP) time and present the results of a stroke quality improvement project (QIP) conducted by Mayo Clinic Arizona's stroke center. We conducted a systematic literature search of Ovid MEDLINE(R), Ovid EMBASE, Scopus, and Web of Science for studies that evaluated DTP time reduction strategies. Those determined eligible for the purpose of this analysis were assessed for quality. The strategies for DTP time reduction were categorized on the basis of modified Target: Stroke Phase III recommendations and analyzed using a meta-analysis. The Mayo Clinic QIP implemented a single-call activation system to reduce DTP times by decreasing the time from neurosurgery notification to case start. Fourteen studies were selected for the analysis, consisting of 2277 patients with acute ischemic stroke secondary to large-vessel occlusions. After intervention, all the studies showed a reduction in the DTP time, with the pooled DTP improvement being the standardized mean difference (1.37; 95% confidence interval, 1.20-1.93; τ2=1.09; P<.001). The Mayo Clinic QIP similarly displayed a DTP time reduction, with the DTP time dropping from 125.1 to 82.5 minutes after strategy implementation. Computed tomography flow modifications produced the largest and most consistent reduction in the DTP time. However, the reduction in the DTP time across all the studies suggests that any systematic protocol aimed at reducing the DTP time can produce a beneficial effect. The relative novelty of mechanical thrombectomy and the consequential lack of research call for future investigation into the efficacy of varying DTP time reduction strategies.


Consolidating biallelic SDHD variants as a cause of mitochondrial complex II deficiency.
Isolated mitochondrial complex II deficiency is a rare cause of mitochondrial respiratory chain disease. To date biallelic variants in three genes encoding mitochondrial complex II molecular components have been unequivocally associated with mitochondrial disease (SDHA/SDHB/SDHAF1). Additionally, variants in one further complex II component (SDHD) have been identified as a candidate cause of isolated mitochondrial complex II deficiency in just two unrelated affected individuals with clinical features consistent with mitochondrial disease, including progressive encephalomyopathy and lethal infantile cardiomyopathy. We present clinical and genomic investigations in four individuals from an extended Palestinian family with clinical features consistent with an autosomal recessive mitochondrial complex II deficiency, in which our genomic studies identified a homozygous NM_003002.3:c.[205 G > A];[205 G > A];p.[(Glu69Lys)];[(Glu69Lys)] SDHD variant as the likely cause. Reviewing previously published cases, these findings consolidate disruption of SDHD function as a cause of mitochondrial complex II deficiency and further define the phenotypic spectrum associated with SDHD gene variants.


Strain-specific quantification of probiotic Bacillus subtilis in feed by imaging high-performance thin-layer chromatography.
The European Union has banned the use of antibiotic growth promoters in animal production, which has led to increased use of probiotic microorganisms. These feed additives result in higher costs for farmers, which is why the demand for a quality control system to quantify probiotics in feeds has increased in recent years. Imaging high-performance thin-layer chromatography (HPTLC) was proven to be a robust method for determining the probiotic Bacillus subtilis DSM 29784 strain based on the production of selective bacterial metabolites and thus characteristic metabolite pattern. However, to quantify the specific probiotic strain in the feed, identification of a strain-specific metabolite not produced by genetically very similar bacteria is necessary. Compared to five bacteria with high genetic similarity, a strain-specific metabolite was formed in the probiotic bacteria by a two-step cultivation procedure. Among others, antimicrobial properties were found for this metabolite, which indicated probiotic activity. The hyphenation of normal-phase HPTLC with reversed-phase high-performance liquid chromatography diode array detection and high-resolution mass spectrometry allowed the preliminary assignment of this strain-specific metabolite to the molecular formula C35H44N6O2 (580.3527 Da). This metabolite, produced each time via an upstream cultivation process to generate the standard levels, was used for the quantification of probiotic active cells in the feed. Data on selectivity, linearity, detection limit, recovery, and precision have shown the good performance of the method.


Rectovaginal fistulas. Therapeutic options.
The management of RVF depends on size, location, and cause; anal sphincter function and overall health status of the patient; and the skill and judgment of the surgeon. Careful preoperative assessment of the fistula, surrounding tissues, and anal sphincter and exclusion of associated disease are essential. With thorough evaluation, thoughtful consideration of treatment options, and meticulous operative technique, patients can be assured of an optimal outcome. Success in treatment of patients with RVF should be measured not just in terms of successful closure of the fistula but also in terms of patient satisfaction with postoperative anal continence.


Evaluation of cold plasma for decontamination of molds and mycotoxins in rice grain.
Few reports are on the application of cold plasma (CP) for mold and mycotoxin control in grain. The aim of this study was to evaluate the viability of CP for mycotoxin and mold control in rice grain. Rice grains artificially contaminated with molds or mycotoxins were subjected to CP treatment. The microbial activities of Aspergillus niger, Rhizopus oryzae, Penicillium verrucosum and Fusarium graminearum were significantly inhibited by CP treatment. The maximal reduction of DON and OTA were 61.25 % and 55.64 %, respectively. The electrical conductivity and malondialdehyde content in rice grain increased at most by 30.14 % and 103.27 %, respectively. The seed germination reduced significantly when treatment time reached 8 min. The major nutrients of rice grain were not affected except for prolamine, which was generally consistent with the results of electron microscopy and Fourier-transform infrared analysis. Cold plasma is a promising method for mold and mycotoxin control in rice grain.


Evidence supporting extracorporeal shock wave therapy for acute and chronic soft tissue wounds.
Objective. Soft tissue wound healing is a complex and well-orchestrated sequence of events on multiple biological levels involving systemic, cellular, and molecular signals. The physiological process of wound healing leads to full tissue repair and regeneration with nearly complete restoration of tissue integrity and functionality. Wounds, particularly among the elderly population, can show delayed or disturbed healing; however, delayed or disturbed healing is also evident in patients with comorbidities such as diabetes, atherosclerosis, venous/arterial insufficiency, reduced mobility due to chronic infirmity, and hypercholesterolemia. Chronic wounds are conditions, which include a wide range of inflammatory and degenerative conditions of the musculoskeletal system. Management of chronic, difficult to heal, or non-healing soft tissue wounds requires a multidisciplinary approach. Often these treatment options have inconsistent and irregular outcomes. Poor response or failure to conservative treatments places a substantial burden on patients, their families, the healthcare system, and society in general. Therefore, the development of a new, effective method of treatment to improve healing of problematic wounds and reduce treatment-related costs is extremely valuable. One such therapy is Extracorporeal Shockwave Therapy (ESWT). ESWT acts through mechanotransduction, which produces therapeutic benefits through complex biological pathways including neovascularization and tissue regeneration in the therapeutic target. Published data thus far suggest that the application of ESWT for soft tissue indications is safe, reliable, cost-effective, and clinically efficacious. The exact biological effects of ESWT on human cells are not completely understood, but are currently undergoing further study. The aim of this review is to provide a general overview of shockwave therapy and its role in the treatment of acute and chronic soft tissue wounds. .


The medical home experience among children with sickle cell disease.
While a large body of research documents acute care services for children with sickle cell disease (SCD), little is known about the primary care experiences of this population. The goal of this study was to determine to what extent children with SCD experienced care consistent with a patient-centered medical home (PCMH). We collected and analyzed data from 150 children, ages 1-17 years, who received care within a large children's hospital. The primary dependent variable was access to a PCMH or its four individual components (regular provider, comprehensive care, family-centered care, and coordinated care) as determined by parental report. Multivariate logistic regression was conducted to investigate associations between socio-demographic variables and having access to a PCMH. Only 11% (16/150) of children qualified as having a PCMH, achieving the required thresholds in all four components. Approximately half of children had access to two or fewer components. Over 90% of children were reported to have a personal provider. Two-thirds of children had access to comprehensive care. Almost 60% of children were reported to receive family-centered care. Only 20% of children had access to coordinated care. No consistent associations were found between socio-demographic variables and having access to a PCMH or its individual components. Within our study sample, children with SCD experienced multiple deficiencies in having access to a PCMH, particularly with respect to care coordination. However, further studies with larger samples are needed to determine associations between socio-demographic variables and having a PCMH.


Thermodynamics of the human brain.
The human brain may be regarded as an irreversible system which is constrained by a fixed inflow of free energy in the form of chemical nourishment from within the body and information from the environment. The evolution of the internal spontaneous process may be described as a path on the saddle surface of entropy production rate in the configuration space of the process variables. From any initial state of high entropy production the system evolves towards the saddle point by a series of regessions to temporary minima alternating with fluctuations which introduce new internal constraints and open new channels for regression. The spontaneous regression steps of the process are to be associated with the learning process and the deductive processes of thought since information is necessarily being stored, whereas the fluctuation or nucleation steps are to be associated with the inductive or creative part of the thought process. The state of consciousness is to be associated with the system undergoing regression. If, under certain boundary conditions, the system attains the stable saddle point, a state of unconsciousness is attained in which no change of state variables occurs in time. The stability of this state is indicated by its resistance to perturbation.


Calculated vibrational properties of pigments in protein binding sites 2: Semiquinones in photosynthetic proteins.
[QA- - QA] Fourier transform infrared difference spectra have previously been obtained using purple bacterial reaction centers from Rhodobacter sphaeroides with unlabeled, 18O and 13C isotope labeled phylloquinone (PhQ, also known as vitamin K1) incorporated into the QA protein binding site (Breton, (1997), Proc. Natl. Acad. Sci. USA94 11318-11323). The nature of the bands in these spectra and the isotope induced band shifts are poorly understood, especially for the phyllosemiquinone anion (PhQ-) state. To aid in the interpretation of the bands in these experimental spectra, ONIOM type QM/MM vibrational frequency calculations were undertaken. Calculations were also undertaken for PhQ- in solution. Surprisingly, both sets of calculated spectra are similar and agree well with the experimental spectra. This similarity suggests pigment-protein interactions do not perturb the electronic structure of the semiquinone in the QA binding site. This is not found to be the case for the neutral PhQ species in the same protein binding site. PhQ also occupies the A1 protein binding site in photosystem I, and the vibrational properties of PhQ- in the QA and A1 binding sites are compared and shown to exhibit considerable differences. These differences probably arise because of changes in the degree of asymmetry of hydrogen bonding of PhQ- in the A1 and QA binding sites.


Oxidative tailoring reactions catalyzed by nonheme iron-dependent enzymes: streptorubin B biosynthesis as an example.
Tailoring enzymes catalyze reactions that modify natural product backbone structures before, during, or after their biosynthesis to create a final product with specific biological activities. Such reactions can be catalyzed by a myriad of different enzyme families and are responsible for a wide variety of transformations including regio- and/or stereospecific acylation, alkylation, glycosylation, halogenation, and oxidation. Within a broad group of oxidative tailoring enzymes, there is a rapidly growing family of nonheme iron- and oxygen-dependent enzymes that catalyze a variety of remarkable hydroxylation, desaturation, halogenation, and oxidative cyclization reaction in the biosynthesis of several important metabolites, including carbapenems, penicillins, cephalosporins, clavams, prodiginines, fosfomycin, syringomycin, and coronatine. In this chapter, we report an expedient method for analyzing tailoring enzymes that catalyze oxidative cyclization reactions in prodiginine biosynthesis via expression of the corresponding genes in a heterologous host, feeding of putative biosynthetic intermediates to the resulting strains, and liquid chromatography-mass spectrometry analyses of the metabolites produced.


Cloning and functional characterization of three flavonoid O-glucosyltransferase genes from the liverworts Marchantia emarginata and Marchantia paleacea.
Flavonoid glycosides are important plant secondary metabolites with broad pharmacological activities. Flavonoid glycosides are generated from aglycones, in reactions catalyzed by typical uridine diphosphate-dependent glycosyltransferases (UGTs). Liverworts produce various types of flavonoid glycosides; however, only two UGTs have been characterized from liverworts to date. Here, we isolated three genes encoding UGTs (MeUGT1, MeUGT2, and MpalUGT1) from the liverwort species Marchantia emarginata and Marchantia paleacea through transcriptome sequencing. Recombinant MeUGT1, MeUGT2, and MpalUGT1 proteins heterologously produced in Escherichia coli exhibited catalytic activity towards multiple flavonoids. MeUGT1 and MpalUGT1 catalyzed the glycosylation of flavonols into the corresponding 3-O-glucosides with UDP-glucose as the sugar donor, while MeUGT2 exhibited a wider substrate specificity that included flavonols, flavones, and flavanones. When MeUGT2 was expressed in E. coli, the yield of flavonol 3-O-glucosides reached to 40-60% with feeding of the substrates kaempferol or quercetin under optimal conditions. Furthermore, heterologous expression of MeUGT1 in Arabidopsis thaliana increased the flavonol glycoside contents in the plants. Therefore, the UGTs characterized in this study could provide new data that will be useful for examining flavonoid biosynthesis in liverworts.


Acute injuries of the diaphragm. An analysis of 165 cases.
The records of 165 patients with diaphragmatic injuries seen at Detroit Receiving Hospital from July 1980 through June 1985 were reviewed. Causes included gunshot wounds in 89 patients, stab wounds in 65, and blunt trauma in 11. Mortality rates were 18%, 5%, and 27%, respectively, deaths being caused primarily by bleeding from associated injuries. Operations on these patients included laparotomy in 123 (75%), thoracotomy in four (2%), and both in 38 (23%). The mortality rates for these operations were 0%, 50%, and 53%, respectively. A presumptive preoperative diagnosis of diaphragmatic injury from chest x-ray findings was possible in only 24 (15%) patients. Of 42 thoracotomies, five were performed in the emergency department for cardiac arrest, with three (60%) deaths. Of 37 thoracotomies performed in the operating room, 17 were for thoracic injuries with six (35%) deaths and 20 were for resuscitation for abdominal injuries with 13 (65%) deaths. In most patients, the diaphragmatic injury was critical only in warning the surgeon that severe injuries might be present in both the chest and abdomen. Of 43 patients admitted with a blood pressure of less than 70 mm Hg or four or more associated injuries, 22 died. However, even in these high-risk patients, if resuscitation raised the initial operating room blood pressure to more than 70 mm Hg, reduced the shock time to less than 30 minutes, and kept blood loss below 10 units, the mortality was only 8% (1/12). In contrast, if none of these conditions could be met, the mortality in this high-risk group was 100% (16/16). Thus more aggressive resuscitative efforts and earlier control of bleeding seem to provide the best chance for improved survival.


Insertion and deletion mutants of vaccinia virus.
Thirteen viable insertion mutants of vaccinia virus have been constructed. These mutants, containing coding sequences of the herpes simplex virus thymidine kinase (HSV-TK) gene, were generated by marker transfer via in vivo recombination. The mutants were identified using a replica filter plating technique by in situ hybridization using 32P-nick translated HSV-TK sequences and obtained as pure cultures by repeated plaque purification. Some of these insertion mutants were in turn used as substrates to generate viable deletion mutants of vaccinia virus in the presence of 5'-bromodeoxyuridine. An example of this approach resulting in a vaccinia virus deleted of approximately 1.5 kb of nonessential DNA is presented. Furthermore, the analysis of spontaneously occurring viable deletion mutants of vaccinia lacking approximately 21.4 kb of nonessential DNA is described.