LPTD: a novel linear programming-based topology determination method for cryo-EM maps.
Topology determination is one of the most important intermediate steps toward building the atomic structure of proteins from their medium-resolution cryo-electron microscopy (cryo-EM) map. The main goal in the topology determination is to identify correct matches (i.e. assignment and direction) between secondary structure elements (SSEs) (α-helices and β-sheets) detected in a protein sequence and cryo-EM density map. Despite many recent advances in molecular biology technologies, the problem remains a challenging issue. To overcome the problem, this article proposes a linear programming-based topology determination (LPTD) method to solve the secondary structure topology problem in three-dimensional geometrical space. Through modeling of the protein's sequence with the aid of extracting highly reliable features and a distance-based scoring function, the secondary structure matching problem is transformed into a complete weighted bipartite graph matching problem. Subsequently, an algorithm based on linear programming is developed as a decision-making strategy to extract the true topology (native topology) between all possible topologies. The proposed automatic framework is verified using 12 experimental and 15 simulated α-β proteins. Results demonstrate that LPTD is highly efficient and extremely fast in such a way that for 77% of cases in the dataset, the native topology has been detected in the first rank topology in <2 s. Besides, this method is able to successfully handle large complex proteins with as many as 65 SSEs. Such a large number of SSEs have never been solved with current tools/methods. The LPTD package (source code and data) is publicly available at https://github.com/B-Behkamal/LPTD. Moreover, two test samples as well as the instruction of utilizing the graphical user interface have been provided in the shared readme file. Supplementary data are available at Bioinformatics online.


Ribavirin monotherapy for the treatment of chronic hepatitis E in an immunosuppressed patient.
We herein report the case of a 54-year-old male patient with a human immunodeficiency virus 1 (HIV-1) infection, usually with low viral loads and CD4 cells < 200-100/mm3 due to thymic exhaustion. He was referred to our clinic because of hypertransaminasemia and cholestasis of a duration of 58 months and liver cirrhosis on FibroScan® without esophageal varices. Nonspecific manifestations included weight loss. Liver disease stigmata and generalized amyotrophy were also present.


Overexpression of TIMP-1 and Sensitivity to Topoisomerase Inhibitors in Glioblastoma Cell Lines.
The multifunctional protein - tissue inhibitor of metalloproteinases-1 (TIMP-1) - has been associated with a poor prognosis in several types of cancers including glioblastomas. In addition, TIMP-1 has been associated with decreased response to chemotherapy, and especially the efficacy of the family of topoisomerase (TOP) inhibitors has been related to TIMP-1. As a second line treatment of glioblastomas, the vascular endothelial growth factor (VEGF) antibody bevacizumab is administered in combination with the TOP1 inhibitor irinotecan and glioblastoma cell levels of TIMP-1 could therefore potentially influence the efficacy of such treatment. In the present study, we aimed to investigate whether a high TIMP-1 expression in glioblastoma cell lines would affect the sensitivity to TOP inhibitors, and whether TIMP-1 overexpressing cells would have alterered growth and invasion. We established TIMP-1 overexpressing subclones from two human glioblastoma cell lines. TIMP-1 overexpressing U87MG cells were significantly more resistant than low TIMP-1 expressing clones and parental cells when exposed to SN-38 (TOP1 inhibitor) or epirubicin (TOP2 inhibitor). No significant differences were observed for the TIMP-1 transfected A172 cells. Implantation of both U87MG and A172 spheroids into organotypic brain slice cultures revealed a reduced growth of TIMP-1 overexpressing U87MG spheroids, however, no significant differences in invasion were observed. The present study suggests that TIMP-1 overexpression reduces the effect of TOP inhibitors in glioblastoma. TIMP-1 also appeared to reduce spheroid growth, but did not influence invasion. Whether TIMP-1 plays a role in irinotecan resistance and has a predictive potential in glioblastoma patients remains to be elucidated.


The Effects of Motor Imagery on Pain and Range of Motion in Musculoskeletal Disorders: A Systematic Review Using Meta-Analysis.
In recent years, there has been an increase in the use of motor imagery (MI) in the rehabilitation of musculoskeletal pain conditions. Across the literature, most reviews have yet to consider Laterality Judgement Task training as a form of MI method. This review aimed to evaluate the effectiveness of using MI as an adjunct to standard rehabilitation on the improvement of pain and range of motion parameters when managing patients with musculoskeletal pain conditions. Searches of 8 major electronic databases were conducted. Data for pain and range of motion were extracted. Meta-analyses (where possible) with either a fixed or random-effect(s) model, standardized mean differences (SMDs), and tests of heterogeneity were performed. Eight clinical controlled trials were identified and included in the meta-analyses. When compared with standard rehabilitation alone, the adjunctive role of MI provided superior pain relief (pooled SMD, -2.25; 95% confidence interval, -4.11 to -0.4; P=0.02), and greater improvement in range of motion (pooled SMD, 3.04; 95% confidence interval, 0.66-5.43; P=0.01) in chronic musculoskeletal pain disorders. The results suggest that MI may be effective for pain relief and improvement in range of motion among chronic musculoskeletal pain conditions, although conclusion is based on a limited certainty of evidence as assessed using the GRADES (Grading of Recommendation, Assessment, Development and Evaluation) approach.


Cluster self-assembly of di[gold(I)]halonium cations.
Treatment of gold(I) halide complexes of the type L-Au-X [where L = PPh(3), PEt(3) with X = Cl, Br, I, or L = 2,6-(MeO)(2)C(6)H(3)PPh(2) with X = Cl] with AgSbF(6) in the molar ratio 2:1 in dichloromethane/tetrahydrofuran at -78 degrees C affords high yields of di[gold(I)]halonium salts of the formula [X[Au(PR(3))](2)](+) SbF(6)(-) (2-8). A determination of the crystal structures of the four triarylphosphine complexes (2-4, 8) revealed the presence of novel tetranuclear dications with a highly symmetrical structure (point group S(4)) that arises from self-assembly of the dinuclear monocations through a set of four equivalent aurophilic Au-Au interactions. A comparison with two reference structures of corresponding chloronium perchlorate and bromonium tetrafluoroborate salts with monomeric, dinuclear cations shows that the geometry of the latter is greatly altered on dimerization to optimize the interactions between the closed-shell metal centers (Au: 5d(10)). Weak metallophilic bonding clearly becomes significant only in crystal lattices where anions with a larger ionic radius (SbF(6)(-) vs. BF(4)(-), ClO(4)(-)) reduce the otherwise dominant role of strong interionic Coulomb forces. The results indicate that aurophilic bonding is indeed an ubiquitous, quite dependable mode of intermetallic interactions provided that the right environment is chosen to allow the weak forces to become operative.


The influence of anodic helical design on fluid flow and bioelectrochemical performance.
In this study three different tubular helical anode designs are compared, for each helical design the pitch and nominal sectional area/liquid flow channel between the helicoids was varied and this produced maximum power densities of 11.63, 9.2 and 6.73Wm(-3) (small, medium and large helical flow channel cross-sections). It is found that the level of mixing and the associated shear rates present in the anodes affects both the power development and biofilm formation. The small helical flow channel carbon anode produced 40% more biofilm and this result was related to modelling data which determined a system shear rate of 237s(-1), compared to 52s(-1) and 47s(-1) for the other reactor configurations. The results from computational fluid dynamic modelling further distinguishes between convective flow conditions and supports the influence of helical structure on system performance, so establishing the importance of anodic design on the overall electrogenic biofilm activity.


Effects of Aerobic Exercise during Pregnancy on 1-Month Infant Neuromotor Skills.
To determine the effects of exercise during pregnancy on the neuromotor development of 1-month-old offspring. We hypothesized that aerobic exercise during pregnancy would be associated with higher neuromotor scores in infants at 1 month of age, based on standard pediatric assessment of neuromotor skills. Seventy-one healthy, pregnant women between 18 and 35 yr were randomly assigned to either aerobic exercise intervention or no exercise (control) group. Women in the exercise group performed 50 min of moderate-intensity, supervised aerobic exercise, three times per week; those in control group maintained usual activity. Neuromotor skills were measured at 1 month of age using the Peabody Developmental Motor Scales, 2nd Edition (PDMS-2). Unpaired t-tests were used to compare infants' PDMS-2 subtest percentiles, Gross Motor Quotients, and Gross Motor Quotient percentile between groups. Infants of women in the exercise group had higher PDMS-2 scores on four of the five variables analyzed relative to infants of nonexercisers. Female infants tended to have improved scores relative to male infants of controls; this difference was attenuated in infants of exercisers. Exercise during pregnancy can positively influence developing systems allowing for improved neuromotor development, thus leading to infants who are more adept at movement, and presumably more likely to be active. Because physical activity is a modifiable risk factor of childhood obesity, these findings suggest that exercise during pregnancy may potentially reduce childhood risk of obesity.


The effects of dynamic loading on intracalcaneal wood implants and on the tissues surrounding them.
The dorsal parts of the calcanei of rabbits were resected and replaced with similarly-shaped implants made of ash wood. In each case the Achilles tendon was reinserted into the projecting part of the implant so as to subject the latter to the dynamic loads generated by use of the limb. The implants were removed from some of the animals after 5 weeks and from the others after 14 weeks. They were evaluated by preparing undecalcified microtome sections and ground sections which were examined by light microscopy and microradiography. Some of the animals underwent vital staining so as to produce polychromatic sequential labelling of the calcaneal specimens. In the area of insertion of the Achilles tendon soft tissue grew into the larger pores of the wood and, in some cases, differentiated into cartilage. Every implant remained stably anchored in the calcaneus. In every animal the intracalcaneal part of the implant became surrounded by new bone which was in direct contact with the surface of the implant and which also formed in the pores of the wood, even all rabbits loaded the operated foot. The formation of new bone in and around the implants demonstrates the basic feasibility of using ash wood as an isoelastic implant material in bone.


Increased interaction and procedural flexibility favoured participation: Study across European cohorts of preterm born individuals.
To understand participation and attrition phenomena variability in European cohorts of individuals born preterm through in-depth exploration of the interplay of situational elements involved. Multi-situated qualitative design, using focus groups, semi-structured interviews and collaborative visual methodology with a purposive sample of adults born preterm, parents and professionals (n = 124) from eight cohorts in seven European countries. Most cohort participants were motivated by altruism/solidarity and gratitude/sense of duty to reciprocate (only absent in adults aged 19 - 21), followed by expectation of direct benefit to one's health and knowledge amongst participating adults. Common deterrents were perceived failure in reciprocity as in insufficient/inadequate interaction and information sharing, and postal questionnaires. Combining multipurpose, flexible strategies for contact and assessment, reminders, face-to-face and shorter periodicity and not simply adding retention strategies or financial incentives favoured participation. Professionals' main challenges entailed resources, funding and, European societal changes related to communication and geopolitical environment. Retention would benefit from tailoring inclusive strategies throughout the cohorts' life cycle and consistent promotion of reciprocal altruistic research goals. Investing in regular interaction, flexibility in procedures, participant involvement and return of results can help mitigate attrition as well as considering mothers as main facilitators to participating children and impaired adults.


Electronic structure and thermochemical properties of silicon-doped lithium clusters Li(n)Si0/+, n = 1-8: new insights on their stability.
A theoretical investigation on small silicon-doped lithium clusters Li(n)Si with n = 1-8, in both neutral and cationic states is performed using the high accuracy CCSD(T)/complete basis set (CBS) method. Location of the global minima is carried out using a stochastic search method and the growth pattern of the clusters emerges as follows: (i) the species Li(n)Si with n ≤ 6 are formed by directly binding one Li to a Si of the smaller cluster Li(n-1)Si, (ii) the structures tend to have an as high as possible symmetry and to maximize the coordination number of silicon. The first three-dimensional global minimum is found for Li(4)Si, and (iii) for Li(7)Si and Li(8)Si, the global minima are formed by capping Li atoms on triangular faces of Li(6)Si (O(h)). A maximum coordination number of silicon is found to be 6 for the global minima, and structures with higher coordination of silicon exist but are less stable. Heats of formation at 0 K (Δ(f)H(0)) and 298 K (Δ(f)H(298)), average binding energies (E(b)), adiabatic (AIE) and vertical (VIE) ionization energies, dissociation energies (D(e)), and second-order difference in total energy (Δ(2)E) of the clusters in both neutral and cationic states are calculated from the CCSD(T)/CBS energies and used to evaluate the relative stability of clusters. The species Li(4)Si, Li(6)Si, and Li(5)Si(+) are the more stable systems with large HOMO-LUMO gaps, E(b), and Δ(2)E. Their enhanced stability can be rationalized using a modified phenomenological shell model, which includes the effects of additional factors such as geometrical symmetry and coordination number of the dopant. The new model is subsequently applied with consistency to other impure clusters Li(n)X with X = B, Al, C, Si, Ge, and Sn.


Morphological parameters of middle cerebral arteries associated with aneurysm formation.
The objective of this work was to investigate the correlation between morphological parameters of the MCA and the formation of aneurysms. MCA aneurysms were diagnosed in 122 cases using CT angiography (including 30 cases of M1 proximal aneurysms, 70 cases of M1 bifurcation aneurysms, and 22 cases of distal aneurysms). Images from these cases were retrospectively compared with images from 50 healthy controls. Morphological parameters including the angle of the MCA with the ICA (α) and the ACA (β) were evaluated in the three aneurysm groups and the control group; parent-daughter angles (γ1, γ2), bifurcation angles (γ3), bifurcation diameters, angle ratios, and branch diameter ratios were also compared between the bifurcation aneurysm group and the control group. The blood vessel parameters between the aneurysm groups and controls were analyzed statistically. There was no statistically significant difference in α between the three groups of aneurysms and the control group (P = 0.381). In comparing β between the three groups of aneurysms and the control group, statistically significant differences were only observed between the MCA distal aneurysm group and the control group (P = 0.010). Compared with the control group, MCA bifurcation aneurysms were associated with larger γ3 and smaller γ1 and γ2 (P < 0.001). This resulted in significantly larger angle ratios in the MCA bifurcation aneurysm group (P < 0.001). For the diameter measurements, the bifurcation diameter of the MCA bifurcation aneurysms was significantly smaller (P = 0.001). The formation of MCA aneurysms is related to morphological parameters.


Comparison of polyphenol, anthocyanin and antioxidant capacity in four varieties of Lonicera caerulea berry extracts.
Four varieties of Lonicera caerulea berries--'Wild', 'Beilei', 'No. 1', and 'No. 2'--were compared with respect to extraction yield, fruit weight, total soluble solids, polyphenol and anthocyanin contents, oxygen radical absorbance capacity (ORAC), and anthocyanin composition. Sixteen individual anthocyanins were identified in the selected varieties. Acylated anthocyanins, cyanidin 3-acetylhexoside and peonidin 3-acetylhexoside, were identified in L. caerulea berries for the first time. Cyanidin-3-glucoside was the most prominent anthocyanin in all four tested varieties. Wild type of L. caerulea fruit ('Wild'), with the highest polyphenol content, contained 14 anthocyanins and the highest ORAC value. Eleven anthocyanins were found in 'Beilei' berries, which had a higher ORAC value than 'No. 1' and 'No. 2'. The highest total soluble solid content and extraction yield were found in 'No. 2' and 'Wild' berries, respectively.


Functional coupling of a mu opioid receptor to G proteins and adenylyl cyclase: modulation by chronic morphine treatment.
A cloned mu opioid receptor was used to study its coupling to signal transduction pathways and its involvement in morphine-induced opioid dependence in stably transfected Chinese hamster ovary (CHO) cells. Membrane binding assays with a mu-selective agonist [(3)H]DAMGO showed that one cell line expresses a high level of mu opioid receptors with a B(max) of approximately 630 fmol/mg membrane protein and a K(d) of 0.47 nM for DAMGO. Stimulation of the transfected cells with DAMGO led to an increase in the low K(m) GTPase activity, indicative of activation of guanine nucleotide regulatory proteins (G proteins), and this effect was blocked by the opioid antagonist naloxone. In addition, binding of the mu opioid receptor to DAMGO was affected by GTP and nonhydrolizable GTP analogs, Gp(NH)pp and GTP-gamma-S. These results suggest a functional coupling between the mu opioid receptor and G proteins. Furthermore, DAMGO treatment of the cells produced a dose-dependent inhibition of the intracellular cyclic adenosine monophosphate (cAMP) level, with an EC(50) value of approximately 30 nM. Chronic treatment of the cells with morphine not only elevated the basal and forskolin-stimulated cAMP levels after morphine withdrawal, but also increased the extent of the DAMGO-induced reduction of intracellular cAMP levels. The whole cell binding assay with [(3)H]DAMGO, on the other hand, did not detect receptor down-regulation after chronic morphine treatment. These results suggest that chronic morphine treatment may trigger a compensatory mechanism in cellular signaling pathways to offset the inhibitory input from the mu receptor without down-regulation of the surface receptor number, and that withdrawal of chronic inhibition leads to elevated activities of adenylyl cyclase to provide a basis for system sensitization.


Does green finance restrain corporate financialization?
Green financial policy is one of the important policy tools for China to implement the national carbon peak goal and carbon neutral vision through financial means. This policy has an important impact on the business strategy of corporates. Based on the data of listed corporates from 2013 to 2020, this study examines the impact mechanism of China's green financial reform and innovation pilot zones (GFRIPZ) on corporate financialization (CF) using the difference-in-difference method. The results show the following: (1) The implementation of GFRIPZ significantly restrains the CF. (2) GFRIPZ reversed the short-sighted behavior of firms and guided them to accelerate the green transformation and upgrading for long-term development. Firms' environmental capital expenditure and research and development expenditure increased significantly. (3) The restraining effect of GFRIPZ on CF is stronger in state-owned firms, firms with low-degree managerial myopia, and high-polluting firms. The research clearly identifies the causal relationship and mechanism between GFRIPZ and CF and reveals the formation mechanism and solution path of CF from the green finance perspective. In addition, this study has implications for guiding the green transformation of entity firms and stopping firms from deviating from their intended purpose.


Ganciclovir-induced apoptosis in HSV-1 thymidine kinase expressing cells: critical role of DNA breaks, Bcl-2 decline and caspase-9 activation.
Although ganciclovir (GCV) is most often used in suicide anticancer gene therapy, the mechanism of GCV-induced cell killing and apoptosis is not fully understood. We analysed the mechanism of apoptosis triggered by GCV using a model system of CHO cells stably transfected with HSV-1 thymidine kinase (HSVtk). GCV-induced apoptosis is due to incorporation of the drug into DNA resulting in replication-dependent formation of DNA double-strand breaks and, at later stages, S and G2/M arrest. GCV-provoked DNA instability was likely to be responsible for the observed initial decline in Bcl-2 level and caspase-9/-3 activation. Further decline in the Bcl-2 level was due to cleavage of the protein by caspase-9, as demonstrated by use of caspase inhibitors and transfection with trans-dominant negative caspase expression vectors. Bcl-2 cleavage resulted in the appearance of a pro-apoptotic 23 kDa Bcl-2 fragment and in excessive cytochrome c release, dephosphorylation of BAD, cleavage of PARP and finally DNA degradation. Since Fas/CD95 and caspase-8 were only slightly activated we conclude GCV-induced apoptosis to occur in this cell system mainly by activating the mitochondrial damage pathway. This process is independent of p53 for which the cells are mutated. Caspase-9 mediated cleavage of Bcl-2 accelerates the apoptotic process and may explain the high potential of GCV to induce apoptosis. Data are also discussed as to implications for HSVtk gene therapy utilizing GCV.


Early radiation-induced liver toxicities are associated with poor survival in patients with advanced hepatocellular carcinoma.
Little is known about the impact of radiotherapy associated early hepatic toxicities. This study is to investigate the risk factors and outcome of early radiation-induced liver disease (early-RILD) in patients with hepatocellular carcinoma. One hundred patients with advanced hepatocellular carcinoma receiving hepatic radiotherapy were included in this retrospective analysis. All had no evidence of intrahepatic tumor progression within 3 months after initiating radiotherapy. The toxicities were graded according to the Common Terminology Criteria for adverse events version 4.0. Early-RILD was defined as any detectable events of RILD occurring during or within 2 weeks after the ending of radiotherapy. Patient- and radiotherapy-related data, and several staging/scoring parameters were retrieved for analysis. Logistic regression analysis was used to find risk factors for early-RILD. Cox regression model was performed to explore prognosticators for survival. Child-Turcotte-Pugh (CTP) score >5 was the predictor for early-RILD (odds ratio 5.38, P = 0.004). The incidence of early-RILD in patients with CTP scores 6/7 and 5 was 34% and 13.2%, respectively. Early-RILD and a Cancer of the Liver Italian Program (CLIP) score > 2 were the two prognostic factors associated with inferior overall survival (hazard ratio 2.79, P = 0.04; hazard ratio = 3.79, P = 0.04, respectively). The median overall survival for patients with early-RILD was 3.5 months compared with 12.7 months in those without this event. The occurrence of early-RILD is associated with high mortality. A CTP score >5 is the most informative factor predicting early-RILD.


Genetic characterization of human immunodeficiency virus type 1 from Beira, Mozambique.
In this report, we examined the genetic diversity of HIV-1 strains circulating in the city of Beira, the second largest metropolitan area in Mozambique. A total of 131 blood samples, collected between August and October 2003 from antiretroviral-naïve individuals, were characterized with a combined approach consisting of heteroduplex mobility assay (HMA) subtyping for gag (n=74) and/or env (n=117) genes, and DNA sequence analysis of proviral env (C2V3C3, n=52), LTR (n=30) and/or pol (n=43) genomic regions. Aside from the identification, by bootscanning analysis, of a viral strain with a C/A1 mosaic C2V3C3 structure, classified as subtype A by env HMA, phylogenetic inference studies of the sequence data demonstrated the circulation of genetically diverse subtype C viruses, predominantly of the R5 type. Inspection of the LTR sequences revealed a pattern of structural and regulatory elements typical of subtype C, with 63.3% of the viruses showing three NF-kappaB binding sites. Analysis of the predicted protease sequences enabled us to detect a single primary mutation (I84V, n=1) associated with resistance to protease inhibitors (PI), while secondary mutations were highly prevalent, some of them in combinations which may confer PI resistance. Although an unexpectedly high rate (11.6%) of reverse transcriptase key mutations (V75A, K103N, Y181C, M184I, or P236L) was detected in the sequences analyzed, our data suggest the non-epidemic circulation of resistant viruses, and the absence of multi-class drug resistant viral strains.


Incidence of Heterotopic Ossification with NSAID Prophylaxis Is Low After Open and Arthroscopic Hip Preservation Surgery.
We evaluate the incidence of heterotopic ossification (HO) development with nonsteroidal anti-inflammatory drug (NSAID) prophylaxis in patients after open and arthroscopic hip preservation surgery. A retrospective review identified patients who underwent hip preservation surgery at a single institution within the past 3 years. Patients who underwent hip arthroscopy with or without periacetabular osteotomy (PAO) or femoral osteotomy (FO) were included. Those who did not receive 3-month postoperative radiographs were excluded. The incidence and Brooker classification (BC) of HO in patients taking Naproxen or another NSAID (meloxicam, celecoxib, indomethacin, or aspirin alone) was assessed using AP radiographs available from 3-, 6-, and 12-month follow-up appointments. Univariate analysis was conducted to compare numerical means and categorical data (significance level P = .05). A total of 328 hips (284 patients) were included. All patients received hip arthroscopy, while 71 patients (21.6%) received concurrent periacetabular osteotomy (PAO; n = 65) or femoral osteotomy (FO; n = 6). Overall, 276 hips (84.4%) received Naproxen for HO prophylaxis. In total, 5 of 328 hips (1.5%) developed HO (4, BC I; 1, BC III). The rate of HO development was significantly higher in males versus females (4 of 121 (3.31%) vs 1 of 207 (.48%), P = .0441). All 5 patients received arthroscopic cam resection and labral repair, and 1 patient also received PAO. Three patients in the Naproxen group (.91%) developed HO, which was not statistically different from those taking a different NSAID (.61%, P = .1797). The incidence of HO development was low with NSAID prophylaxis after hip preservation surgery.


Recent insights into kidney diseases associated with glomerular cysts.
Glomerular cysts can exist in the context of several different kidney diseases. Advances in the last few years have begun to unravel the genetic bases and pathogenesis of some of these entities, many of which have an origin in abnormal development. In this review, we highlight recent insights into three types of disease associated with glomerular cysts: (1) mutations of the hepatocyte nuclear factor 1beta (HNF-1beta)in the recently described renal cysts and diabetes syndrome, (2) mutations of OFD1in the oral facial digital syndrome type 1 and (3) the role of fetal urinary tract obstruction.


Multi-wavelength anomalous diffraction using medium-angle X-ray solution scattering (MADMAX).
Proteins are dynamic molecules whose function in virtually all biological processes requires conformational motion. Direct experimental probes of protein structure in solution are needed to characterize these motions. Anomalous scattering from proteins in solution has the potential to act as a precise molecular ruler to determine the positions of specific chemical groups or atoms within proteins under conditions in which structural changes can take place free from the constraints of crystal contacts. In solution, anomalous diffraction has two components: a set of cross-terms that depend on the relative location of the anomalous centers and the rest of the protein, and a set of pure anomalous terms that depend on the distances between the anomalous centers. The cross-terms are demonstrated here to be observable and to provide direct information about the distance between the anomalous center and the center of mass of the protein. The second set of terms appears immeasurably small in the context of current experimental capabilities. Here, we outline the theory underlying anomalous scattering from proteins in solution, predict the anomalous differences expected on the basis of atomic coordinate sets, and demonstrate the measurement of anomalous differences at the iron edge for solutions of myoglobin and hemoglobin.


Truncated TALE-FP as DNA Staining Dye in a High-salt Buffer.
Large DNA molecules are a promising platform for in vitro single-molecule biochemical analysis to investigate DNA-protein interactions by fluorescence microscopy. For many studies, intercalating fluorescent dyes have been primary DNA staining reagents, but they often cause photo-induced DNA breakage as well as structural deformation. As a solution, we previously developed several fluorescent-protein DNA-binding peptides or proteins (FP-DBP) for reversibly staining DNA molecules without structural deformation or photo-induced damage. However, they cannot stain DNA in a condition similar to a physiological salt concentration that most biochemical reactions require. Given these concerns, here we developed a salt-tolerant FP-DBP: truncated transcription activator-like effector (tTALE-FP), which can stain DNA up to 100 mM NaCl. Moreover, we found an interesting phenomenon that the tTALE-FP stained DNA evenly in 1 × TE buffer but showed AT-rich specific patterns from 40 mM to 100 mM NaCl. Using an assay based on fluorescence resonance energy transfer, we demonstrated that this binding pattern is caused by a higher DNA binding affinity of tTALE-FP for AT-rich compared to GC-rich regions. Finally, we used tTALE-FP in a single molecule fluorescence assay to monitor real-time restriction enzyme digestion of single DNA molecules. Altogether, our results demonstrate that this protein can provide a useful alternative as a DNA stain over intercalators.


Urinary felinine excretion in intact male cats is increased by dietary cystine.
Felinine is a branched-chain sulfur amino acid present in the urine of certain Felidae, including domestic cats. The objective of the present study was to determine if additional cystine and/or dietary N would increase felinine and N-acetylfelinine excretion by intact male cats fed a low-protein(LP) diet. Feeding five adult intact male cats an LP diet (18.8% of metabolisable energy (ME) as protein) v. a high-protein diet (38.6% of ME as protein) resulted in a trend (P=0.08) for decreased urinary felinine and no change in N-acetylfelinine excretion. In a 23 d study, when the LP diet was supplemented with L-cystine at 9.3 g/kg DM, urinary felinine:creatinine ratio showed a linear two-fold (121 %) increase (P<0.01) from 0.24 (SEM 0.05) to 0.53 (SEM 0.13) after 10 d. Subsequent feeding of the LP diet resulted in a decrease in felinine excretion to base levels. Plasma gamma-glutamyl felinylglycine concentrations were consistent with the excretion of felinine. Supplementation of the LP diet with L-cystine (9.3 g/kg DM),dispensable amino acids and arginine to a second group (n 5) also resulted in a significant (P<0.01) but smaller (+72 %) increase in the daily felinine:creatinine ratio (0.25 (SEM 0.04) to 0.43 (SEM 0.05)). The degree of felinine N-acetylation within groups was unaffected by dietary addition and withdrawal of amino acids. The results indicate that felinine synthesis is regulated by cystine availability, and that arginine may be physiologically important in decreasing felinine biosynthesis in intact male cats.


Improvement of the diagnosis of the cause of pleural effusion in patients with lung cancer by simultaneous quantification of carcinoembryonic antigen (CEA) and neuron-specific enolase (NSE) pleural levels.
Carcinoembryonic antigen (CEA) and neuron-specific enolase (NSE) level determinations were carried out by radioimmunoassay in pleural fluid and plasma samples obtained from 24 patients with malignant pleural effusions and 18 patients with non-malignant pleural effusions, and compared to cytological and pathological results. Using a pathological cut-off level of 25 ng/ml for CEA and 8 ng/ml for NSE, we demonstrated that, in the diagnosis of the malignant nature of pleural effusions, the simultaneous quantification of CEA and NSE in pleural fluid possesses better discriminative values than the simultaneous quantification of both markers in plasma or the separate quantification of each marker, in pleural fluid and in plasma.


Complete Genomic and Ultrastructural Analysis of a Nam Dinh Virus Isolated from Culex pipiens quinquefasciatus in China.
The Nam Dinh virus (NDiV) was isolated from Culex quinquefasciatus in Shenzhen, China, for the first time, in 2011. In this study, we characterized the ultrastructure of NDiV, determined its complete genome sequence and made comparisons with other known nidoviruses. Electron microscopic observation revealed that the NDiV strain isolated in China produced viral nucleocapsid-like particles and vesicles in host cells. The extracellular virions were enveloped and were spherical with short spikes. The complete genome sequence of the newly isolated NDiV was submitted to the GenBank database (GenBank accession number KF522691). Sequencing of the viral genome showed that the homologies of NDiV isolated in China and Vietnam were greater than 94.0% and 89.0% at the nucleotide and amino acid sequence levels, respectively. Moreover, gene substitution was detected, whereas insertions and deletions were not. A phylogenetic tree analysis showed that these viruses belong to the genus Alphamesonivirus1 of the family Mesoniviridae. The similarity between the two viruses regarding morphological and molecular biological characteristics indicates that the molecular genetics of NDiV are conservative and that the regional differences are unlikely to have a significant effect. This is the first report of the isolation and complete sequencing of a mesonivirus in mainland China.


Listening to familiar music induces continuous inhibition of alpha and low-beta power.
How the brain responds temporally and spectrally when we listen to familiar versus unfamiliar musical sequences remains unclear. This study uses EEG techniques to investigate the continuous electrophysiological changes in the human brain during passive listening to familiar and unfamiliar musical excerpts. EEG activity was recorded in 20 participants while they passively listened to 10 s of classical music, and they were then asked to indicate their self-assessment of familiarity. We analyzed the EEG data in two manners: familiarity based on the within-subject design, i.e., averaging trials for each condition and participant, and familiarity based on the same music excerpt, i.e., averaging trials for each condition and music excerpt. By comparing the familiar condition with the unfamiliar condition and the local baseline, sustained low-beta power (12-16 Hz) suppression was observed in both analyses in fronto-central and left frontal electrodes after 800 ms. However, sustained alpha power (8-12 Hz) decreased in fronto-central and posterior electrodes after 850 ms only in the first type of analysis. Our study indicates that listening to familiar music elicits a late sustained spectral response (inhibition of alpha/low-beta power from 800 ms to 10 s). Moreover, the results showed that alpha suppression reflects increased attention or arousal/engagement due to listening to familiar music; nevertheless, low-beta suppression exhibits the effect of familiarity.NEW & NOTEWORTHY This study differentiates the dynamic temporal-spectral effects during listening to 10 s of familiar music compared with unfamiliar music. This study highlights that listening to familiar music leads to continuous suppression in the alpha and low-beta bands. This suppression starts ∼800 ms after the stimulus onset.


Black-white preterm birth disparity: a marker of inequality.
The racial disparity in preterm birth (PTB) is a persistent feature of perinatal epidemiology. Its consistency is not only an outcome of reproductive interest but may function as a persistent inequality to which women are exposed over time. Herein we sought to analyze exposure to area-level PTB disparity and its association with individual-level PTB risk. Using geocoded birth records (1999-2001) and year 2000 census data for Wake County, North Carolina, we created a tract-level PTB disparity variable for each woman in our study area, comprising all births in her tract excluding her birth. Multilevel random intercept models for exposure to neighborhood PTB disparity and odds of PTB were fit. Exposure to area-level PTB disparity was associated with increased odds of PTB for all women. After adjustment for maternal and neighborhood covariates, odds of delivering preterm increased by 25%, 26%, and 36% for the second, third, and fourth quartiles of exposure to PTB disparity (odds ratio [OR]=1.25, 95% confidence interval [CI]: 1.00-1.57; OR=1.26, 95% CI: 1.01-1.58; and OR=1.36, 95% CI: 1.06-1.75, respectively). Exposure to persistent inequality, in the form of PTB disparity, appears associated with individual-level PTB risk. Knowing where PTB disparity is greatest has implications for PTB prevention policy efforts to buffer adverse conditions.


Correlation of the pediatric metabolic index with NAFLD or MAFLD diagnosis, and serum adipokine levels in children.
Non-alcoholic fatty liver disease (NAFLD) is characterized by ectopic fat deposition in the liver. However, a recent classification of this condition, which also integrates the presence of coexisting metabolic disorders, termed Metabolic dysfunction Associated Fatty Liver Disease (MAFLD), has been proposed. NAFLD is increasingly common in early childhood, partly due to the increase in metabolic disease in this age. Thus, studying hepatic steatosis in the metabolic context has become important in this population as well. However, NAFLD, and thus MAFLD, diagnosis in children is challenging by the lack of non-invasive diagnostic tools comparable to the gold standard of hepatic biopsy. Recent studies have reported that the Pediatric Metabolic Index (PMI) could be a marker of insulin resistance and abnormal liver enzymes, but its association with NAFLD, MAFLD, or altered adipokines in these conditions has not been reported. The aim of this study is to evaluate the correlation between PMI with the diagnosis of NAFLD or MAFLD, together with serum levels of leptin and adiponectin, in school-age children. A cross sectional study was carried out in two hundred and twenty-three children without medical history of hypothyroidism, genetic, or chronic diseases. Anthropometry, liver ultrasound, and serum levels of lipids, leptin, and adiponectin were evaluated. The children were classified as having NAFLD or non-NAFLD, and a subgroup of MAFLD in the NAFLD group was analyzed. The PMI was calculated by the established formulas for age and gender. PMI correlated positively with the presence and severity of NAFLD (r = 0.62, p<0.001 and r = 0.79, p<0.001 respectively) and with the presence of MAFLD (r = 0.62; p<0.001). Also, this index correlated positively with serum leptin levels (r = 0.66; p<0.001) and negatively with serum adiponectin levels (r= -0.65; p<0.001). PMI showed to be a good predictor for diagnosing NAFLD in school-age children when performing a ROC curve analysis (AUROC=0.986, p< 0.0001). PMI could be a useful tool for the early diagnosis of NAFLD or MAFLD in children. However, future studies are necessary to establish validated cut-off points for each population.


Transcranial direct current stimulation in treatment resistant depression: a randomized double-blind, placebo-controlled study.
Anodal transcranial direct current stimulation (tDCS) of the prefrontal cortex has been proposed as therapeutic intervention in major depression. According to clinical needs, this study addresses the question whether tDCS is effective in treatment resistant major depressive episodes. Twenty-two patients with a major depressive episode were randomly assigned to a cross-over protocol comparing tDCS and placebo stimulation add-on to a stable antidepressant medication. The parameters of active tDCS were: 1 or 2 mA for 20 minutes/day, anode over the left dorsolateral prefrontal cortex, cathode over the contralateral supraorbital region. Active and placebo tDCS was applied for 2 weeks using indistinguishable DC stimulators. Patients, raters, and operators were blinded to treatment conditions. There was no significant difference in depression scores after 2 weeks of real compared with 2 weeks of sham tDCS. Scores on the Hamilton Depression Rating Scale were reduced from baseline by 14.7% for active tDCS and 10% for placebo tDCS. In contrast, subjective mood ratings showed an increase in positive emotions after real tDCS compared with sham tDCS. Anodal tDCS, applied for 2 weeks, was not superior to placebo treatment in patients with treatment resistant depression. However, secondary outcome measures are pointing to a positive effect of tDCS on emotions. Therefore, modified and improved tDCS protocols should be carried out in controlled pilot trials to develop tDCS towards an efficacious antidepressant intervention in therapy-resistant depression.


Increased levels of carboxyhemoglobin and serum iron as an indicator of increased red cell turnover in preeclampsia.
Patients with severe preeclampsia are reported to have microangiopathic hemolytic anemia. This study demonstrates that increased red cell turnover with heme catabolism is also common in mild preeclampsia. Heme catabolism results in equimolar production of carboxyhemoglobin, iron, and bilirubin. A concomitant rise in these constituents of venous blood would support this hypothesis. Patients with antepartum preeclampsia had mean carboxyhemoglobin concentrations (2.72% total hemoglobin) greater than those of control patients (0.65%) (p less than 0.001) and serum iron concentrations (98.5 micrograms/dl) greater than those of control patients (66.1 micrograms/dl) (p less than 0.01). Bilirubin concentrations were not different. Post partum, carboxyhemoglobin and iron concentrations returned toward normal (1.38% and 50.2 micrograms/dl, respectively). Disparity in the magnitude of increase of heme catabolites produced in equimolar proportion is explained by differences in the kinetics of clearance. The data are most consistent with increased destruction of maternal red cells, even in mild preeclampsia. Potential implications of elevated carboxyhemoglobin on maternal and fetal oxygenation are discussed.


Type II neurofibromatosis presenting as quadriceps atrophy.
A young woman aged 26 years presented with atrophy of the left quadriceps progressing over one year. Magnetic resonance imaging (MRI) showed a large lesion of the lumbar plexus compatible with neurinoma. Cerebral MRI revealed a lesion in the right eighth cranial nerve also compatible with neurinoma. On further questioning of the patient, it was learned that her mother had undergone surgery twice for acoustic neurinoma. Type II neurofibromatosis was diagnosed from the basis of clinical picture, neuroimaging findings and family history. This case is unusual for the lack of central nervous system (CNS) symptoms. Expansive radicular lesions compatible with neurinoma should prompt cerebral MRI. Regular examination of family members at risk and early diagnosis can decrease the high mortality associated with this condition.


Becoming a Caregiver: Experiences of Young Adults Moving into Family Caregiving Roles.
While the vast majority of family caregivers struggle to find balance between different roles in their lives, young adult caregivers are faced with the atypical challenge of caring for family member while simultaneously accomplishing developmental tasks typical of this stage in life (e.g., establishing career, developing romantic relationships). This exploratory, qualitative study examined strategies used by young adults to adopt family caregiving roles. These strategies can be described as embracement, compromise, and integration. While each approach allowed for the young adult to facilitate their caregiving role, additional research is needed to understand how the strategy affects the emerging adult's development.


B-Raf and the inhibitors: from bench to bedside.
The B-Raf protein is a key signaling molecule in the mitogen activated protein kinase (MAPK) signaling pathway and has been implicated in the pathogenesis of a variety of cancers. An important V600E mutation has been identified and can cause constitutive B-Raf activation. Recent studies have evaluated a variety of small molecule inhibitors targeting B-Raf, including PLX4032/vemurafenib, dabrafenib, LGX818, GDC0879, XL281, ARQ736, PLX3603 (RO5212054), and RAF265. Therapeutic resistance has been identified and various mechanisms described. This review also discussed the current understanding of B-Raf signaling mechanism, methods of mutation detection, treatment strategies as well as potential methods of overcoming therapeutic resistance.


TCH-165 attenuates cardiac ischaemia/reperfusion injury by balancing mitochondrial dynamics via increasing proteasome activity.
The proteasome is the main complex responsible for maintaining intracellular protein homeostasis, impairment of which is associated with cardiac ischaemia/reperfusion (I/R) injury. The small molecule TCH-165 has been found to activate the 20S proteasome to remove disordered proteins in multiple myeloma and glioblastoma. However, the preventive effect of TCH-165 against I/R-mediated cardiac impairment in mice remains largely unknown. Here, a cardiac I/R model was established in mice. Heart function was assessed with echocardiography. Cardiac infarction, myocyte death, and superoxide level were evaluated by 2,3,5-triphenyltetrazolium chloride (TTC)-Evans blue staining, terminal deoxynucleotidyl transferase-mediated dUTP nick and labelling (TUNEL) assay and immunostaining, respectively. Our results showed that TCH-165 treatment markedly ameliorated I/R-mediated cardiac dysfunction and decreased the infarct size, apoptosis, and superoxide levels. Mechanistically, TCH-165 increased immunoproteasome subunit expression/activity, increasing pro-fission protein dynamin-1-like protein (DNM1L, also known as DRP1) degradation and the expression of the pro-fusion proteins mitofusin 1/2 (Mfn1/2) and thereby leading to mitochondrial fission/fusion balance. In vitro experiments confirmed that inhibition of proteasome activity by epoxomicin abolished the protective effect of TCH-165 against hypoxia/reoxygenation (H/R)-induced increases in cardiomyocyte apoptosis, superoxide production and mitochondrial fission. In summary, TCH-165 is a newly discovered inducer of immunoproteasome activity that exerts a preventive effect against cardiac I/R damage by targeting Drp1 degradation, indicating that it may be as a potential therapeutic candidate for ischaemic heart disease.


Phylogenetic reconstruction and shell evolution of the Diplommatinidae (Gastropoda: Caenogastropoda).
The fascinating and often unlikely shell shapes in the terrestrial micromollusc family Diplommatinidae (Gastropoda: Caenogastropoda) provide a particularly attractive set of multiple morphological traits to investigate evolutionary patterns of shape variation. Here, a molecular phylogenetic reconstruction, based on five genes and 2700 bp, was undertaken for this family, integrated with ancestral state reconstruction and phylogenetic PCA of discrete and quantitative traits, respectively. We found strong support for the Diplommatininae as a monophyletic group, separating the Cochlostomatidae into a separate family. Five main clades appear within the Diplommatininae, corresponding with both coiling direction and biogeographic patterns. A Belau clade (A) with highly diverse (but always sinistral) morphology comprised Hungerfordia, Palaina, and some Diplommatina. Arinia (dextral) and Opisthostoma (sinistroid) are sister groups in clade B. Clade C and D solely contain sinistral Diplommatina that are robust and little ornamented (clade C) or slender and sculptured (clade D). Clade E is dextral but biogeographically diverse with species from all sampled regions save the Caroline Islands. Adelopoma, Diplommatina, Palaina, and Hungerfordia require revision to allow taxonomy to reflect phylogeny, whereas Opisthostoma is clearly monophyletic. Ancestral state reconstruction suggests a sinistral origin for the Diplommatinidae, with three reversals to dextrality.


Performance constraints in decathletes.
Physical performance by vertebrates is thought to be constrained by trade-offs between antagonistic pairs of ecologically relevant traits and between conflicting specialist and generalist phenotypes, but there is surprisingly little evidence to support this reasoning. Here we analyse the performance of world-class athletes in standardized decathlon events and find that it is subject to both types of trade-off, after correction has been made for differences between athletes in general ability across all 10 events. These trade-offs may have imposed important constraints on the evolution of physical performance in humans and other vertebrates.


Sociodemographic factors and environmental workers' knowledge of the impact of awareness creation on sustainable disposal of solid wastes.
This study examined sociodemographic factors and environmental workers' knowledge of the impact of awareness-creation on the sustainable disposal of solid waste in Cross River State, Nigeria. To achieve the purpose of the study, two research questions and two hypotheses guided the study. The sequential explanatory mixed-method design was adopted in the study. The population for the study was 331 respondents, consisting of 120 staff and 211 government-approved service providers from the Cross-River State Ministry of Environment. The population also comprised the sample size of the study. The total enumeration sampling technique was employed. A 42-item structured questionnaire titled "Questionnaire Instrument on Sociodemographic Factors and Environmental Workers' Knowledge of the Impact of Awareness Creation on Sustainable Disposal of Solid Wastes" (QISFEWKIACSDSW) and a Focus Group Discussion Schedule (FGDS) were the instruments used for data collection. The instruments were face validated by three experts: two from Social Science Education and one from the Measurement and Evaluation Unit of the Department of Science Education, University of Nigeria, Nsukka. The Cronbach Alpha Reliability Method was used to determine the internal consistency of the instrument. Data collected from the questionnaire was analyzed using mean and standard deviation for the research questions while independent sampled t-test statistic was used to test the null hypotheses at a 0.05 level of significance and the FGDS was used to supplement the findings of the quantitative analysis. Findings of the study revealed that, to a high extent, awareness creation based on location and gender can contribute positively to the sustainable disposal of solid waste materials. Based on the findings of the study, it was recommended, among others, that enlightenment programs such as awareness creation, town hall meetings, and workshops be organized among community members on the need for sustainable disposal of solid waste materials in Cross River State, Nigeria.


Preserved memory capacities in aged Lou/C/Jall rats.
Although memory impairments are a hallmark of aging, the degree of deficit varies across animal models, and is likely to reflect different states of deterioration in metabolic and endocrinological properties. This study investigated memory-related processes in young (3-4 months) and old (24 months) Sprague-Dawley rats (SD), which develop age-linked pathologies such as obesity or insulin-resistance and Lou/C/Jall rats, which do not develop such impairments. In short- and long-term memory recognition tasks, old Lou/C/Jall rats were never impaired whereas old SD rats were deficient at 1 and 24h latencies. The expression of N-methyl-d-aspartate receptors (NMDAR)-mediated synaptic plasticity in CA1 hippocampal networks shifted towards lower activity values in old Lou/C/Jall rats whereas long-term potentiation was impaired in age-matched SD rats. Age-related decrease in NR2A subunits occurred in both strains, extended to NR2B, NR1 and GluR1 subunits in older animals (28 months) but only in SD rats. Therefore, the Lou/C/Jall rats can be considered as a model of healthy aging, not only in terms of its preserved metabolism, but also in terms of cognition and synaptic plasticity.


Implementation of an emergency department back pain clinical management tool on the early diagnosis and testing of spinal epidural abscess.
Spinal epidural abscess (SEA) is a rare, catastrophic condition for which diagnostic delays are common. Our national group develops evidence-based guidelines, known as clinical management tools (CMT), to reduce high-risk misdiagnoses. We study whether implementation of our back pain CMT improved SEA diagnostic timeliness and testing rates in the emergency department (ED). We conducted a retrospective observational study before and after implementation of a nontraumatic back pain CMT for SEA in a national group. Outcomes included diagnostic timeliness and test utilization. We used regression analysis to compare differences before (January 2016-June 2017) and after (January 2018-December 2019) with 95% confidence intervals (CIs) clustered by facility. We graphed monthly testing rates. In 59 EDs, pre versus post periods included 141,273 (4.8%) versus 192,244 (4.5%) back pain visits and 188 versus 369 SEA visits, respectively. After implementation, SEA visits with prior related visits were unchanged (12.2% vs. 13.3%, difference +1.0%, 95% CI -4.5% to 6.5%). Mean number of days to diagnosis decreased but not significantly (15.2 days vs. 11.9 days, difference -3.3 days, 95% CI -7.1 to 0.6 days). Back pain visits receiving CT (13.7% vs. 21.1%, difference +7.3%, 95% CI 6.1% to 8.6%) and MRI (2.9% vs. 4.4%, difference +1.4%, 95% CI 1.0% to 1.9%) increased. Spine X-rays decreased (22.6% vs. 20.5%, difference 2.1%, 95% CI -4.3% to 0.1%). Back pain visits receiving erythrocyte sedimentation rate or C-reactive protein increased (1.9% vs. 3.5%, difference +1.6%, 95% CI 1.3% to 1.9%). Back pain CMT implementation was associated with an increased rate of recommended imaging and laboratory testing in back pain. There was no associated reduction in the proportion of SEA cases with a related prior visit or time to SEA diagnosis.


Intraoperative graft flow profiles in coronary artery bypass surgery: A meta-analysis.
Conduits used in coronary artery bypass artery grafting (CABG) have different properties and flow profiles. We compared intraoperative mean graft flow (MGF) between arterial and venous conduits, off-pump CABG (OPCABG) and on-pump CABG (ONCABG) procedures, skeletonized and pedicled internal mammary artery (IMA) grafts, and pulsatility index (PI) between OPCABG and ONCABG, in pairwise meta-analyses. Following a systematic literature search, all studies comparing MGF in arterial and venous grafts, were included. The primary endpoint was comparison of pooled MGF between arterial and venous grafts. Secondary endpoints were comparisons of pooled MGF in OPCABG vs ONCABG, anastomosed skeletonized vs pedicled IMA grafts, free skeletonized vs pedicled IMA grafts and PI in OPCABG versus ONCABG. A total of 25 studies with 4443 patients were included. Compared with venous grafts, arterial grafts had lower MGF (standardized mean difference [SMD], -0.28; 95% confidence interval [CI, -0.34; -0.22]; P < .001). OPCABG was associated with significantly lower MGF compared to ONCABG (SMD, -0.29; 95%CI, -0.50; -0.08]; P = .01). No differences were found in MGF between skeletonized vs pedicled IMA after anastomosis (SMD, 0.32; 95%CI [-0.08; 0.71]; P = .11) or in free flow (SMD, 0.76; 95%CI [-0.14; 1.65]; P = .10). No difference was found in PI between OPCABG and ONCABG. At meta-regression, age was associated with higher MGF, while OPCABG was associated with lower MGF. Intraoperative flow of venous conduits is higher than that of arterial grafts. Compared to OPCABG surgery, graft flow is higher in ONCABG. In skeletonized and pedicled IMA conduits, no difference in flow profiles was found.


[The significance of immunological analysis for the detection of residual neuroblasts in bone marrow].
Immunological analysis is complementary to morphological investigation to detect bone marrow (BM) metastases in neuroblastoma patients. It is essential at time of BM harvest for an autograft, since it is the only examination which allows a precise evaluation of the graft contamination by malignant cells. Simple indirect immunofluorescence staining with anti-neuroblastoma monoclonal antibodies (UJ13A and HSAN 1-2) allows a final detection of about 1% malignant cells in the BM, and 1% when cells are gathered in clumps. The use of an immunocytochemical method (alkaline phosphatase) together with double immunofluorescence staining permits to detect as few as one residual neuroblastoma cell in 10(4) normal ones. These two methods used together have allowed to assess the neuroblastoma nature of rare isolated cells with pseudo-lymphocytic aspect.


Preparation of Internal Standards for 2D-UPLC-MS/MS Quantification of Noncanonical DNA Bases.
Reliable quantitative analysis of DNA modification using liquid chromatography coupled with tandem mass spectrometry requires stable isotope-labeled internal standards. Only some of them are commercially available. Here we present a method allowing for the synthesis of [13C10,15N2]-5-methyl-2'-deoxycytidine from [13C10,15N2]-2'-deoxythymidine. We also describe an approach for the oxidation of [13C10,15N2]-5-methyl-2'-deoxycytidine and [13C10,15N2]-2'-deoxythymidine with Na2S2O8, leading to the generation of [13C10,15N2]-5-formyl-2'-deoxycytidine, [13C10,15N2]-5-carboxy-2'-deoxycytidine or [13C10,15N2]-5-(hydroxymethyl)-2'-deoxyuridine, correspondingly. Moreover, we provide optimized protocols for the oxidation of [13C5,15N2]-thymine to [13C10,15N2]-5-hydroxymethyluracil, [13C10,15N2]-5-formyluracil, and [13C10,15N2]-5-carboxyuracil using Na2S2O8.


Urticarial vasculitis: Clinical and laboratory findings with a particular emphasis on differential diagnosis.
Urticarial vasculitis (UV) is a rare cutaneous vasculitis of small vessels characterized by recurrent episodes of wheal-like lesions that tend to last more than 24 hours, healing with a residual ecchymotic postinflammatory hyperpigmentation. The histopathologic pattern of UV is that of leukocytoclastic vasculitis, consisting of fibrinoid necrosis of dermal vessels' walls and neutrophil-rich perivascular inflammatory infiltrates. Although its etiopahogenesis remains still undefined, UV is now regarded as an immune complex-driven disease with activation of the complement cascade, leading to exaggerated production of anaphylatoxins that are responsible for neutrophil recruitment and activation. This condition can be categorized into 2 main entities according to serum complement levels: normocomplementemic UV and hypocomplementemic UV, the latter being associated with circulating anti-C1q autoantibodies and possible extracutaneous manifestations. Systemic multiorgan involvement may be seen particularly in syndromic hypocomplementemic UV, also known as McDuffie syndrome. This review summarizes the clinicopathological and laboratory features as well as the underlying pathophysiological mechanisms of UV. A focus on its main differential diagnoses is provided, that is, chronic spontaneous urticaria, bullous pemphigoid, IgA (Henoch-Schönlein purpura) and IgM/IgG immune complex vasculitis, lupus erythematous tumidus, Wells syndrome, erythema multiforme, cutaneous mastocytosis, cryopyrin-associated periodic syndromes, and coronavirus disease 2019-associated and anti-severe acute respiratory syndrome coronavirus 2-vaccine-associated urticarial eruptions.


Bias and variability of diagnostic spectral parameters extracted from closing sounds produced by bioprosthetic valves implanted in the mitral position.
A method is proposed to estimate the bias and variability of eight diagnostic spectral parameters extracted from mitral closing sounds produced by bioprosthetic heart valves. These spectral parameters are: the frequency of the dominant (F1) and second dominant (F2) spectral peaks, the highest frequency of the spectrum found at -3 dB (F-3), -10 dB (F-10) and -20 dB (F-20) below the highest peak, the relative integrated area above -20 dB of the dominant peak (RIA20), the bandwidth at -3 dB of the dominant spectral peak (BW3), and the ratio of F1 divided by BW3 (Q1). The bias and variability of four spectral techniques were obtained by comparing parameters extracted from each technique with the parameters of a spectral "standard." This "standard" consisted of 19 normal mitral sound spectra computed analytically by evaluating the Z transform of a sum of decaying sinusoids on the unit circle. Truncation of the synthesized mitral signals and addition of random noise were used to simulate the physiological characteristics of the closing sounds. Results show that the fast Fourier transform method with rectangular window provides the best estimates of F1 and Q1, that the Steiglitz-McBride method with maximum entropy (pole-zero modeling with four poles and four zeros) can best evaluate F2, F-20, RIA20 and BW3, and that the all-pole modeling with covariance method (16 poles) is best suited to compute F-3. It was also shown that both the all-pole modeling and the Steiglitz-McBride methods can be used to estimate F-10. It is concluded that a single algorithm would not provide the best estimates of all spectral parameters.


James Parkinson and his essay on "shaking palsy", two hundred years later.
In 1817, British physician James Parkinson published a 66-page document entitled "Essay on the Shaking Palsy". This brief text became a classical and fundamental piece in the history of medicine and, in particular, of neurology. The authors of this article wish to pay tribute to this great pioneer of neurology, 200 years after the publication of his findings, which would, in turn, immortalize his name and give rise to the renaming on the entity in 1860 by Professor Jean Martin Charcot, father of neurology. It would be known, henceforth as Parkinson's disease.


The challenges of neonatal sepsis management.
To present current evidence on the etiology, risk factors, diagnosis, and management of early and late neonatal sepsis. Non-systematic review of the Medline (PubMed), Scopus, Web of Science, Cochrane, and Google Scholar databases regarding the following terms: neonatal sepsis, early neonatal sepsis, late neonatal sepsis, empirical antibiotic therapy, sepsis calculator, vancomycin, newborn, preterm newborn. Neonatal sepsis is a frequent cause of neonatal morbidity and mortality. Its diagnosis is difficult. Continuous observation of the patient is critical to diagnostic suspicion. When neonatal sepsis is suspected, bacteriological tests should be collected. Vancomycin should not be routinely using in the empirical antibiotic regimen in late neonatal sepsis, and the main protective mechanisms against neonatal sepsis are handwashing and the use of breast milk. Newborns constitute a group that is more vulnerable to sepsis. Knowledge of risk factors and etiological agents allows a better approach to the newborn with sepsis.


Transplantation Using Renal Grafts With Multiple Renal Arteries: A Putative Study on the Impact of Arterial Reconstruction Technique and Site of Implantation on Outcomes.
In the present retrospective study, we analyzed the outcomes of patients transplanted with grafts with multiple renal arteries (MRAs). In total, 89 patients were transplanted with renal grafts with MRAs from 2003 to 2018. Demographic characteristics; type of donor; warm and cold ischemia times; arterial anastomosis technique; complications; graft function at first month, first year, and last outpatient clinic visit; and patient and graft survival were all retrospectively evaluated. The mean age of the patients was 40.4 ± 13.3 years. Fifty-six patients (62.9%) were male. In total, 42 patients (47.2%) received renal grafts from living related donors. In group A (n = 24; 27%), anastomosis was performed separately to the recipient external or internal iliac arteries; in group B (n = 38; 42.7%), the secondary artery was anastomosed to the main artery in a side-to-side fashion to form a single common orifice; in group C (n = 27; 30.3%), secondary arteries were anastomosed to the main renal artery in an end-to-side fashion. Creatinine clearance at the first month was significantly lower for deceased-donor grafts compared to living-donor renal grafts (P < .05). Creatinine clearance in the first postoperative month was significantly lower in group A and creatinine clearance in the first year was significantly lower in group C (P < .05). The best survival was found for anastomosis to the internal iliac artery (P < .05). MRAs can be safely used and the reconstruction technique does not matter if the graft kidney's arterial supply is preserved and the internal iliac artery is chosen for anastomosis.


2,9-Dimethyl-1,10-phenanthrolin-1-ium (6-carb-oxy-4-hy-droxy-pyridine-2-carboxyl-ato-κO,N,O)(4-hy-droxy-pyridine-2,6-dicarboxyl-ato-κO,N,O)zincate(II) 2.35-hydrate: a proton-transfer compound.
In the title compound, (C(14)H(13)N(2))[Zn(C(7)H(3)NO(5))(C(7)H(4)NO(5))]·2.35H(2)O, the Zn(II) atom is coordinated by two N atoms and four O atoms from the carboxyl-ate groups of the 4-hy-droxy-pyridine-2,6-dicarboxyl-ate and 6-carb-oxy-4-hy-droxy-pyridine-2-carboxyl-ate ligands, forming a distored octa-hedral geometry. In the anion, the two pyridine rings are inclined to one another by 87.75 (13)°. Two types of robust O-H⋯O hydrogen bond synthons, viz. R(2) (2)(16) and R(6) (6)(42), link the anions to form a two-dimensional network parallel to the bc plane. Furthermore, O-H⋯O, N-H⋯O, N-H⋯N and weak C-H⋯O hydrogen bonds connect the two dimensional networks, forming a three-dimensional structure. In the crystal, there are also C-H⋯π and π-π inter-actions [centroid-centroid distances of 3.5554 (18) and 3.7681 (18) Å], and C=O⋯π inter-actions [O⋯centroid distance = 3.117 (2) Å] present. One of the three crystal water molecules shows an occupancy of 0.35.


Salt coatings functionalize inert membranes into high-performing filters against infectious respiratory diseases.
Respiratory protection is key in infection prevention of airborne diseases, as highlighted by the COVID-19 pandemic for instance. Conventional technologies have several drawbacks (i.e., cross-infection risk, filtration efficiency improvements limited by difficulty in breathing, and no safe reusability), which have yet to be addressed in a single device. Here, we report the development of a filter overcoming the major technical challenges of respiratory protective devices. Large-pore membranes, offering high breathability but low bacteria capture, were functionalized to have a uniform salt layer on the fibers. The salt-functionalized membranes achieved high filtration efficiency as opposed to the bare membrane, with differences of up to 48%, while maintaining high breathability (> 60% increase compared to commercial surgical masks even for the thickest salt filters tested). The salt-functionalized filters quickly killed Gram-positive and Gram-negative bacteria aerosols in vitro, with CFU reductions observed as early as within 5 min, and in vivo by causing structural damage due to salt recrystallization. The salt coatings retained the pathogen inactivation capability at harsh environmental conditions (37 °C and a relative humidity of 70%, 80% and 90%). Combination of these properties in one filter will lead to the production of an effective device, comprehensibly mitigating infection transmission globally.


Incidence and time course of atrial fibrillation following patent foramen ovale closure.
Determine the true incidence and time course of atrial fibrillation (AF) after patent foramen ovale closure (PFOc) using implantable loop recorders (ILR) placed during cryptogenic stroke evaluation. Published trials report a 2%-6.6% incidence of postimplant atrial fibrillation (PIAF) after PFOc, which is probably a gross underestimation, as only patients presenting in AF were captured. Episodes of paroxysmal and silent AF would have been missed. Of 761 patients who underwent PFOc at a single center between January 2016 and December 2020, 35 patients had an ILR implanted before PFOc, without documentation of AF, and had ≥1 month of monitoring post-PFOc. The incidence, onset, and conclusion of AF episodes were determined from a review of patient records. The mean duration of ILR monitoring was 54.6 ± 39.4 weeks after PFOc. AF occurred in 13/35 (37%) patients. PFOc patients who developed PIAF were older than those who did not (62 ± 11 vs. 52 ± 14 years, p = 0.03). In 12/13, the initial PIAF event occurred within 4 weeks of PFOc, with the greatest frequency around 2 weeks and conclusion by 12 weeks in all. No recurrent strokes occurred during ILR monitoring. The actual incidence of PIAF was far greater than previously reported and was significantly associated with older age at PFOc. The timing of PIAF onset and termination were consistent with a postimplant inflammatory mechanism. The higher actual PIAF incidence underscores its low stroke potential in this population. A larger prospective trial is required to validate these preliminary results.


Dead-Seq: Discovering Synthetic Lethal Interactions from Dead Cells Genomics.
Pooled genetic screens have revolutionized the field of functional genomics, yet perturbations that decrease fitness, such as those leading to synthetic lethality, have remained difficult to quantify at the genomic level. We and colleagues previously developed "death screening," a protocol based on the purification of dead cells in genetic screens, and used it to identify a set of genes necessary for mitochondrial gene expression, translation, and oxidative phosphorylation (OXPHOS), thus offering new possibilities for the diagnosis of mitochondrial disorders. Here, we describe Dead-Seq, a refined protocol for death screening that is compatible with most pooled screening protocols, including genome-wide CRISPR/Cas9 screening. Dead-Seq converts negative-selection screens into positive-selection screens and generates high-quality data directly from dead cells, at limited sequencing costs.


[Systemic candidiasis in medical intensive care unit: analysis of risk factors and the contribution of colonization index].
Description of the epidemiological and clinical characteristics of the patients introducing risk factors of invasive candidiasis. Analysis of risk factors for candidiasis invasive and evaluation of the contribution of colonization index (CI) in the diagnosis of the systematic candidiasis in medical intensive care. Prospective observational study (October 2007 to October 2009). The selected patients present risk factors of system IC candidiasis with an infectious syndrome or clinical signs suggestive of Candida infection and hospitalized more than 48 hours in medical intensive care unit. Pittet's colonization index was calculated at admission and then once a week added to a blood culture. Patients were classified according to level of evidence of Candida infection and the degree of colonization (CI<0.5, CI ≥ 0.5). The study included 100 patients. Mean age of our patients was 55.8 ± 18.2 years with male prevalence. Neurological disease was the most frequent pathology in admission (48%). The most common risk factors were broad-spectrum antibiotics and foreign material. In the various mycology IC specimens, Candida albicans was the most frequent, followed by C. tropicalis, then C. glabrata. The CI was greater than or equal to 0.5 at 53% of the patients, and less than 0.5 in 47% of the cases. Among the patients, 15% developed an invasive candidiasis. In multivariate analysis, the corticosteroid therapy was associated with a high colonisation (IC ≥ 0.5) and neutropenia with a high risk of systemic candidiasis. The positive predictive value of CI was 26%. The negative predictive value was 98%, the sensitivity and specificity was 93% and 48% respectively. CI has the advantage to provide a quantified data of the patient's situation in relation to the colonization. But, it isn't helpful with patients having an invasive candidiasis in medical intensive care unit.


Importance of the natural history of benign prostatic hyperplasia in the evaluation of pharmacologic intervention.
The variable natural history of untreated symptomatic benign prostatic hyperplasia (BPH) and the fact that therapeutic intervention is based more on subjective than objective criteria of progressive disease have important consequences for appropriately evaluating the effects of pharmacologic intervention on clinical BPH. To evaluate accurately the success of drug therapy for symptomatic BPH, any trial must include a placebo treatment arm for comparison. However, for such a placebo control to be legitimate, it must not affect the natural history of BPH. To evaluate the effect of placebo treatment on BPH, a large body of data following the natural history of untreated clinical BPH from a variety of independent studies was combined and compared to those from a large variety of independent studies in which a placebo-treated group of patients with clinical BPH also followed. These comparisons demonstrate that 1) placebo treatment does not affect the natural history of the disease; 2) spontaneous improvement usually occurs within the first 6 months of initial presentation of symptoms, if it is to occur at all; and 3) 3-6 months of follow-up are needed to determine if a patient is going to get worse. Thus, to evaluate accurately the potential benefit of any medical intervention for symptomatic BPH, placebo-controlled clinical trials will be required and should be of at least 6 month's duration.


Free and Substrate-Immobilised Lipases from Fusarium verticillioides P24 as a Biocatalyst for Hydrolysis and Transesterification Reactions.
Fungal enzymes are widely used in technological processes and have some interesting features to be applied in a variety of biosynthetic courses. Here, free and substrate-immobilised lipases from Fusarium verticillioides P24 were obtained by solid-state fermentation using wheat bran as substrate and fungal carrier. Based on their hydrolytic and transesterification activities, the lipases were characterised as pH-dependent in both reactions, with higher substrate conversion in an alkaline environment. Thermally, the lipases performed well from 30 to 45 °C, being more stable in mild conditions. Organic solvents significantly influenced the lipase selectivity using different vegetable oils as fatty acid source. Omega(ω)-3 production in n-hexane achieved 45% using canola oil, against ≈ 18% in cyclohexane. However, ω-6 production was preferably produced for both solvents using linseed oil with significant alterations in the yield (≈ 79% and 49% for n-hexane and cyclohexane, respectively). Moreover, the greatest enzyme selectivity for ω-6 led us to suppose a lipase preference for the Sn1 position of the triacylglycerol. Lastly, a transesterification reaction was performed, achieving 90% of ester conversion in 72 h. This study reports the characterisation and use of free and substrate-immobilised lipases from Fusarium verticillioides P24 as an economic and efficient method for the first time.


Optimization of topical therapy: partitioning of drugs into stratum corneum.
To optimize a topical formulation for therapeutic effect generally implies that the flux of drug into the skin be maximized. This requirement means that the product of drug concentration in the vehicle (Cv) and drug partition coefficient (PC) between stratum corneum (SC) and vehicle be as large as possible. While Cv is a formulation variable which can be easily manipulated up to the drug's saturation solubility, PC is a parameter that is difficult to predict a priori. However, there is no question that an ability to evaluate PC would greatly facilitate the efficient screening of drugs and formulations. We have measured the SC/water and SC/isopropylmyristate (a model lipophilic vehicle) PCs of seven drugs; acitretin, progesterone, testosterone, diazepam, estradiol, hydrocortisone, and caffeine, SC/water PCs were determined as a function of the following variables: (i) initial drug concentration in the vehicle, (ii) length of equilibrium, (iii) SC source and preparation technique, and (iv) SC delipidization. The data obtained were reproducible and physicochemically consistent, and they show that useful partitioning information from both aqueous and nonaqueous vehicles can be obtained with the biological tissue of greatest relevance. The SC/water PCs of the steroids were in reasonable agreement with previous measurements. A facile approach to an integral determinant of formulation optimization is suggested, therefore, by these observations.


Quality of life in patients with vitiligo: a cross-sectional study based on Vitiligo Quality of Life index (VitiQoL).
Vitiligo is a multi-factorial pigmentary skin disorder. Recently, the importance of emotional and psychological issues is proposed in incidence, progression, relapse and remission of vitiligo. There are limited studies conducted in developing countries, which assess life quality of patients with vitiligo. The aim of this study was the application and evaluation of a disease-specific quality of life index in Iranian patients, for the first time. This cross-sectional biphasic study was conducted on 25 patients as a pilot and another 173 patients as the main study group, in Razi Hospital, Tehran, Iran, 2013-2014. Persian version of Vitiligo Quality of Life index (VitiQoL) was developed with backward-forward method. Based on the pilot study, the validity and reliability were assessed. The Vitiligo Area and Score Index (VASI), VitiQoL, and their relationship, demographic and clinical characteristic of patients were measured. The Mean and standard deviation of the VitiQoL score was 30.5 ± 14.5 (range 0-60 in Persian version). There was a significant relationship between VASI score and VitiQoL (p = 0.015, r = 0.187). Confirmatory factor analysis revealed three important factors within VitiQoL: participation limitation, stigma, and behavior. In subscale analysis based on behavior factor, female patients had poorer quality of life (p = 0.02). Concomitant psychiatric problems, e.g. anxiety and depression, were not associated with QOL; however, they were near to being meaningful (p = 0.06, r = 0.14). VitiQoL is a valid index in estimating life quality of vitiligo patients and has proper relation to disease severity. Focusing on patient's life quality is an important entity in the management of vitiligo patients; relevant supportive group-based consultations and therapies are also important arms when approaching vitiligo.


[Protein digestability and amino acid absorption in various segments of the digestive tract of pigs. 1. Methods of animal experimentation, protein content and amino acid patterns of the digesta in the small intestine and in feces after feeding different feeds].
Re-entrance cannulae were applied by way of operations at the end of the small intestine of a larger number of growing pigs. After a post-operative phase these animals were fed with various rations (N-free diet, fattening feed for pigs, rations with dried skim milk resp. wheat gluten + lysine and wheat + wheat gluten + lysine). During the main experiment periods the ileum digesta (24 hour-periods of collecting) and the feces were quantitatively registered. The protein and amino acid content ascertained in the feed rations, the digesta and feces samples are reported, comparatively evaluated and discussed. The amount of amino acids (in mg per kg intake of dry matter) contained in the ileum digesta after N-free feeding were statistically calculated with regard to differences occuring between the animals and days. For most amino acids a significant decrease in the ileum digesta could be detected during the course of the N-free feeding period. Moreover, the amount of amino acids in the ileum digesta was compared with the metabolic fecal amino acids, from which conclusions could be drawn concerning the disappearance rate of the individual amino acids in large intestine.


Ruscogenin ameliorates dasatinib-induced intestinal barrier dysfunction via ErbB4/YAP and ROCK/MLC pathways.
Dasatinib is effective in the treatment of chronic and acute myeloid leukemia, which could cause the side effect of gastrointestinal bleeding by overdose or longtime use. Ruscogenin (RUS) from the traditional Chinese medicine Ophiopogon japonicas could protect endothelial microvascular barrier function. In this study, the therapeutic effect and underlying mechanisms of RUS were investigated on intestinal barrier dysfunction induced by dasatinib. Male C57BL/6 J mice were given three doses of dasatinib (70, 140, 210 mg/kg, ig) and RUS (3, 10, 30 μg/kg, ip) to explore the effect of dasatinib on intestinal barrier and the intervention of RUS. It was proved that dasatinib could reduce intestinal blood flow, inhibit phosphorylation of EGFR family member v-erb-b2 avian erythroblastic leukemia viral oncogene homolog 4 (ErbB4)/YES-associated protein (YAP) and activation of Rho-associated coiled coil-containing protein kinase (ROCK)/phosphorylation of (myosin light chain) MLC. RUS could significantly increase intestinal blood flow, improve intestinal injury, reduce Evans blue leakage and serum content of FITC-dextran 4 kDa, and increase the expression of connexin (ZO-1, Occludin and VE-cadherin). Meanwhile, the in vitro effect of RUS (0.01, 0.1, 1 μM) on the dysfunction of the endothelial barrier was observed in dasatinib (150 nM)-pretreated HUVECs. The results showed that RUS suppressed dasatinib-induced the leakage of Evans blue, and degradation of F-actin and connexin. Furthermore, RUS could significantly increase the phosphorylation of ErbB4 at Tyr1284 site and YAP at Ser397 site, and inhibit ROCK expression and phosphorylation of MLC at Ser19 site in vivo and in vitro. In conclusion, the present research proved that RUS could suppress the side effects of dasatinib-induced intestinal barrier dysfunction by regulating ErbB4/YAP and ROCK/MLC pathways.


Zero-Dimensional MXene-Based Optical Devices for Ultrafast and Ultranarrow Photonics Applications.
In recent years, MXene has become a hotspot because of its good conductivity, strong broadband absorption, and tunable band gap. In this contribution, 0D MXene Ti3C2Tx quantum dots are synthesized by a liquid exfoliation method and a wideband nonlinear optical response from 800 to 1550 nm is studied, which have a larger nonlinear absorption coefficient β of -(11.24 ± 0.14) × 10-2 cm GW-1. The carrier dynamic processes of 0D MXene are explored with ultrahigh time resolution nondegenerate transient absorption (TA) spectroscopy, which indicates that the TA signal reaches its maximum in 1.28 ps. Furthermore, 0D MXene is used to generate ultrashort pulses in erbium or ytterbium-doped fiber laser cavity. High signal-to-noise (72 dB) femtosecond lasers with pulse durations as short as 170 fs with spectrum bandwidth of 14.8 nm are obtained. Finally, an ultranarrow fiber laser based on 0D MXene is also investigated and has a full width at half maximum of only 5 kHz, and the power fluctuation is less than 0.75% of the average power. The experimental works prove that 0D MXene is an excellent SA and has a promising application in ultrafast and ultranarrow photonics.


Development and Implementation of a Semi-Automated Workflow for Point-of-Care Ultrasound Billing and Documentation Within an Electronic Health Record.
Point-of-care ultrasound (POCUS) is widely used for both diagnostic and therapeutic purposes. With its many advantages, including ease of use, real-time multisystem assessment, affordability, availability, and accuracy, it has been adopted by all medical specialties. Despite its advantages, the lack of standard workflow and automated billing solutions makes it difficult to launch a comprehensive POCUS program. In this work, we describe how we created and implemented an efficient standardized EHR-based workflow for POCUS that has been used across multiple division and settings within our organization.


Synthesis of a compound soluble in organic solvents from deoxycytidine triphosphate in permeabilized normal human lymphocytes.
Normal human lymphocytes may be rendered permeable to deoxynucleoside triphosphates. When [3H]dCTP is furnished to permeabilized lymphocytes tow compounds are formed: DNA and a compound soluble in organic solvents. [3H]dCTP incorporation is higher in stimulated lymphocytes than in unstimulated cells. Some characteristics of the system are reported.


Studies on the CEA-like substance in gastric juice.
Concentration and heterogeneity of CEA-like substance in gastric juice were studied using radioimmunoassay. Statistically significant increase of CEA-like substance in gastric juice was found in advanced atrophic gastritis (P less than 0.01), early gastric cancer (P less than 0.05) and advanced gastric cancer (P less than 0.01) as compared with normal subjects. In cases with atrophic gastritis with a high degree of intestinal metaplasia, the concentrations above 300 microgram/dl were noticed. The results indicate that increased concentrations of CEA-like substance in gastric secretions may strongly suggest the presence of a marked intestinal metaplasia and/or cancerous changes of gastric mucosae, including the early cancer. The distribution of CEA activity in gel filtration fractions of gastric juice was compared using kits of two different radioimmunoassay systems. The patterns of CEA activities were different between the two different kits used, but the main peaks were located in the fractions with the molecular weight of 20x10(4) daltons corresponding to that of serum CEA. It is considered, however, that the CEA-like substance in gastric juice specimens may be more or less heterogenous when any of the methods is used.


Three dimensional gait analysis in patients with symptomatic component mal-rotation after total knee arthroplasty.
Purpose of the present cohort study was the determination of lower body function and rotation in patients with symptomatic component mal-rotation after total knee arthroplasty using instrumented 3D gait analysis. A consecutive series of 12 patients (61.3 years ± 11.4 years) were included suffering under remaining pain or limited range of motion at least six months after total knee arthroplasty. A CT-scan according to the protocol of Berger et al. and instrumented 3D gait analysis were carried out including clinical examination, videotaping, and kinematic analysis using a Plug-in Gait model. Outcome variables were temporospatial parameters as well as kinematics in sagittal and transversal plane. Data for reference group were collected retrospectively and matched by age and gender. Temporospatial parameters of the study group showed decreased velocity, cadence, and step length as well as increased step time. Single limb support was reduced for the affected limb. In sagittal plane, maximum knee flexion during swing phase was reduced for the replaced knee joint. In transverse plane, there was hardly any difference between affected and non-affected limb. Compared to the reference group, both limbs show significant increased internal ankle rotation and external hip rotation. There were significant strong linear correlations between ankle rotation and hip rotation as well as ankle rotation and radiological tibial mal-rotation. Patients with symptomatic component mal-rotation after total knee arthroplasty showed typically functional deficits. The affected and non-affected limb showed significant increased internal ankle rotation and external hip rotation, while only the affected, replaced knee showed reduced internal knee rotation. Identification of rotational abnormalities of hip and ankle joints seems to be mandatory in TKA to identify the patient group with external hip rotation, internal ankle rotation, and an elevated risk for symptomatic rotational TKA component mal-alignment.


Preventive use of respiratory support after scheduled extubation in critically ill medical patients-a network meta-analysis of randomized controlled trials.
Respiratory support has been increasingly used after extubation for the prevention of re-intubation and improvement of prognosis in critically ill medical patients. However, the optimal respiratory support method is still under debate. This network meta-analysis (NMA) aims to evaluate the comparative effectiveness of various respiratory support methods used for preventive purposes after scheduled extubation in critically ill medical patients. A systematic database search was performed from inception to December 19, 2019, for randomized controlled trials (RCTs) that compared a preventive use of different respiratory support methods, including conventional oxygen therapy (COT), noninvasive ventilation (NIV), high-flow oxygen therapy (HFOT), and combinational use of HFOT and NIV (HFOT+NIV), after planned extubation in adult critically ill medical patients. Study selection, data extraction, and quality assessments were performed in duplicate. The primary outcomes included re-intubation rate and short-term mortality. Seventeen RCTs comprising 3341 participants with 4 comparisons were included. Compared with COT, NIV significantly reduced the re-intubation rate [risk ratio (RR) 0.55, 95% confidence interval (CI) 0.39 to 0.77; moderate quality of evidence] and short-term mortality (RR 0.66, 95% CI 0.48 to 0.91; moderate quality of evidence). Compared to COT, HFOT had a beneficial effect on the re-intubation rate (RR 0.55, 95% CI 0.35 to 0.86; moderate quality of evidence) but no effect on short-term mortality (RR 0.79, 95% CI 0.56 to 1.12; low quality of evidence). No significant difference in the re-intubation rate or short-term mortality was found among NIV, HFOT, and HFOT+NIV. The treatment rankings based on the surface under the cumulative ranking curve (SUCRA) from best to worst for re-intubation rate were HFOT+NIV (95.1%), NIV (53.4%), HFOT (51.2%), and COT (0.3%), and the rankings for short-term mortality were NIV (91.0%), HFOT (54.3%), HFOT+NIV (43.7%), and COT (11.1%). Sensitivity analyses of trials with a high risk of extubation failure for the primary outcomes indicated that the SUCRA rankings were comparable to those of the primary analysis. After scheduled extubation, the preventive use of NIV is probably the most effective respiratory support method for comprehensively preventing re-intubation and short-term death in critically ill medical patients, especially those with a high risk of extubation failure.


Role of the insulin-like growth factor system on an estrogen-dependent cancer phenotype in the MCF-7 human breast cancer cell line.
We previously established that exposure of the estrogen receptor (ER) alpha positive MCF-7 human breast cancer cell line to 17-beta-estradiol (E2) results in the post-confluent development of multilayered cellular aggregates (foci) which is consistent with the in vivo cancer phenotype of uncontrolled cellular proliferation. In this investigation, the interaction between the insulin-like growth factor receptor (IGF-IR) and ER-signaling systems in regard to post-confluent focus development was studied. We demonstrated that focus development requires the presence of E2 and insulin-like growth factor I (IGF-I) or insulin-like growth factor II (IGF-II), as well as intact ER and IGF-IR. Focus development in MCF-7 cultures, which occurs only after formation of a confluent monolayer, coincides with E2 regulation of key members of the IGF-signaling system such as IGF-IR, IGF-II, insulin receptor substrate 1 (IRS-1), and insulin-like growth factor binding protein 3 (IGFBP-3), as demonstrated by real-time polymerase chain reaction (PCR). To establish the relevancy of an intact IGF-signaling system for foci formation, we generated stable clones from MCF-7 with IGF-IR suppressed by siRNA. Results from these studies implicate signaling through the IGF-IR to be an integral requirement for E2-dependent post-confluent proliferation and focus formation. In summary, these studies establish the interactive roles of IGFs and E2 in the post-confluent development of foci, and will allow subsequent identification of targets for therapeutic intervention in the control and treatment of estrogen-dependent breast cancer.


The Role and Place of Thanatoradiological Studies in the Pathological Examination of Fetuses and Newborns.
Analysis of published data on the possibilities of using postmortem radiation studies in perinatology is carried out and the results of own thanatoradiological studies of the bodies of dead fetuses and newborns are presented. The possibilities of postmortem radiation studies for differential diagnosis of stillborn and deceased newborns, evaluation of the severity of maceration and the time of intrauterine fetal death, detection of pathological changes in the brain and spinal cord, respiratory and digestive organs, in the cardiovascular and urinary systems were demonstrated. It is concluded that postmortem CT has a high diagnostic efficiency in the study of the bone skeleton, free fluid accumulations in serous cavities and gas in the vessels and tissues of dead fetuses and deceased newborns. The advantage of postmortem MRI is more effective visualization of internal organs and soft tissues, which allows assessing their topography and size, as well as identifying a wide range of pathological changes. For a comprehensive objective analysis of the bodies of stillborn and deceased newborns, combined use of both imaging methods (CT and MRI) is required. At the same time, thanatoradiology should be used as a part of a comprehensive pathological study, but not as a substitute for traditional autopsy.


Effect of long-term exposure to aluminum on the acetylcholinesterase activity in the central nervous system and erythrocytes.
Aluminum (Al), a neurotoxic agent, has been associated with Alzheimer's disease (AD), which is characterized by cholinergic dysfunction in the central nervous system. In this study, we evaluated the effect of long-term exposure to aluminum on acetylcholinesterase (AChE) activity in the central nervous system in different brain regions, in synaptosomes of the cerebral cortex and in erythrocytes. The animals were loaded by gavage with AlCl(3) 50 mg/kg/day, 5 days per week, totalizing 60 administrations. Rats were divided into four groups: (1) control (C); (2) 50 mg/kg of citrate solution (Ci); (3) 50 mg/kg of Al plus citrate (Al + Ci), and (4) 50 mg/kg of Al (Al). AChE activity in striatum was increased by 15% for Ci, 19% for Al + Ci and 30% for Al, when compared to control (P < 0.05). The activity in hypothalamus increased 23% for Ci, 26% for Al + Ci and 28% for Al, when compared to control (P < 0.05). AChE activity in cerebellum, hippocampus and cerebral cortex was decreased by 11%, 23% and 21% respectively, for Al, when compared to the respective controls (P < 0.05). AChE activity in synaptosomes was increased by 14% for Al, when compared to control (P < 0.05). Erythrocyte AChE activity was increased by 17% for Al + Ci and 11% for Al, when compared to control (P < 0.05). These results indicate that Al affects at the same way AChE activity in the central nervous system and erythrocyte. AChE activity in erythrocytes may be considered a marker of easy access of the central cholinergic status.


Effect of stress and time for recovery on the amount of compensatory growth after grazing.
We tested the hypothesis that the amount of compensatory growth after defoliation is affected by the level of stress at which plants grow when defoliated and by the length of time for recovery. Growth response to defoliation went from partial compensation when plants were growing at high relative growth rates (RGR) to overcompensation when plants were more stressed and growing at low RGR. Defoliation released plants from the limitation imposed by the accumulation of old and dead tissue and this release overrode the negative effect of biomass loss. Compensatory growth resulted from a higher RGR aboveground that was not associated with a reduction in RGR belowground. Time available for recovery had a major impact on the outcome of defoliation. With a short time for recovery, RGR was decreased by defoliation because an immediate increase in net assimilation rate was overridden by a reduction in the ratio of leaf area to plant weight. After defoliation, this ratio increased quickly due to a larger allocation to leaf growth and lower leaf specific weights, resulting in higher RGR. We conclude that the compensatory response to grazing depends on the type and level of stress limiting growth. Allocation and physiological responses to stress may positively or negatively affect the response to grazing and, simultaneously, grazing may alleviate or aggravate the effects of different types of stress.