Applied biomedical research in mental retardation--prevention.
Prevention of mental retardation is discussed in terms of etiology and level of prevention (primary, secondary or tertiary). The aim of primary preventive medicine is to avert the disease before its onset. Secondary preventive medicine involves early identification and treatment of a disease to reduce its duration and/or severity. Tertiary prevention limits the degree of disability and fosters rehabilitation in chronic diseases. Prevention of mental retardation relies heavily on primary and secondary preventive techniques. Generally, preventive techniques related to organically caused mental retardation are secondary. Attempts are made to identify and treat impairments which cause deficient mental functioning. Since such impairments can occur at any time and are not always observable, routine developmental screening is recommended. A two-stage developmental screening procedure developed by the authors consists of an abbreviated version of the Denver Developmental Screening Test (DDST) followed by the full DDST when indicated. Primary preventive techniques are appropriate for the prevention of environmentally-caused mental retardation. If negative environmental influences can be identified and eliminated early, socio-cultural mental retardation might be averted or if identified early may be decreased to prevent the retardation process. To meet the need for an environmental screening instrument, the authors developed the Home Screening Questionnaire (HSQ). A combined developmental/environmental screening process is recommended.


Determination of cathepsins B, D and G concentration in eutopic proliferative endometrium of women with endometriosis by the surface plasmon resonance imaging (SPRI) technique.
To determine the concentrations of cathepsins B, D and G in proliferative eutopic endometrium of patients with and without endometriosis, by use of the surface plasmon resonance imaging (SPRI) technique. A total of 55 patients were recruited in the study: 31 patients with endometriosis (stages I-IV) and 24 controls. Endometrial samples were obtained in the first phase of the menstrual cycle from regularly menstruating premenopausal women, prior to laparoscopy, by the use of aspiration biopsy. Endometriosis was appropriately classified according to the Revised American Fertility Society classification and confirmed by histopathology in every case. The SPRI technique was used to determine the concentration of cathepsins B, D and G. To compare the two groups for quantitative data, Mann-Whitney-Wilcoxon's test was used due to the non-normal distribution of the tested variables and normality of distribution was assessed using Shapiro-Wilk W test. The concentration of the three examined cathepsins was higher in the proliferative eutopic endometrium of patients with endometriosis, especially in advanced stages, e.g. III and IV, when compared to healthy individuals. Corresponding median values were, for cathepsin B: [7.93 pmol/mg (min-max 2.82-15.71) vs 1.2 pmol/mg (min-max 0.7-15.49) p=0.0014], for cathepsin D: [1.86 pmol/mg (min-max 0.51-5.4) vs 1.03 pmol/mg (min-max 0.4-2.72) p=0.00041] and for cathepsin G: [0.6 pmol/mg (min-max 0.33-2.51) vs 0.3 pmol/mg (min-max 0.16-1.29) p=0.00051]. Increased concentrations of cathepsins B, D and G in the proliferative eutopic endometrium may play a role in the implantation of endometrial tissue outside the uterine cavity.


Effect of legumes in energy reduced dietary approaches to stop hypertension (DASH) diet on blood pressure among overweight and obese type 2 diabetic patients: a randomized controlled trial.
This study aimed to investigate the effects of legumes in dietary approaches to stop hypertension (DASH) on blood pressure and urinary sodium and potassium in participants with type 2 diabetes. We further investigated whether changes in individual dietary food groups in the DASH diet contribute to blood pressure. Participants were randomized to the traditional DASH diet (n = 150) or the legume-based DASH diet (n = 150). Blood pressure and urinary sodium and potassium were measured at baseline and after 16-week interventions. An intention-to-treat approach with multiple imputations of missing data was applied. The restricted cubic spline (RCS) was applied to assess the linearity and explore the shape of the relationship between the changes in food groups and systolic blood pressure (SBP) in the two intervention groups. A reduction in SBP and urinary sodium was observed at week 16 in both dietary interventions, and this reduction was more significant in the legume-based DASH diet, than in the DASH diet. In the legume-based DASH diet, SBP decreased with the legume intake of more than 95.8 g/day (P nonlinear < 0.001). The multivariable RCS analysis showed a U-shaped relationship of refined grains, an L-shaped relationship of fruits, and a linear relationship of sweet, energy, sodium and vegetables intake with SBP. The DASH diet, enriched in legumes, could improve SBP in participants with type 2 diabetes. In DASH diet, the balance of consumption of refined grains and fruits along with higher vegetable consumption and lower sweet, sodium and energy intake reduced the SBP. IRCT20090203001640N17.


Intramedullary tuberculoma of the Spinal Cord, Clinical features & imaging: Possibility of early diagnosis with imaging?
Involvement of vertebral column is common in tuberculosis(TB) but intramedullary tuberculomas are rare. Spinal intramedullary tuberculoma are extremely rare, seen in only 2 of 100,000 cases of TB and 2 of 1000 cases of CNS TB. Intramedullary tuberculomas normally respond well to conventional antituberculous medications, requiring surgery only occasionally. MRI is optimal diagnostic modality in intramedullary tuberculoma as it can show specific findings. This report describes a case of Intramedullary Tuberculoma of the Spinal Cord with clinical features and specific MRI findings with review of litreture.


Cost-Effectiveness of Screening, Surveillance, and Endoscopic Eradication Therapies for Managing the Burden of Esophageal Adenocarcinoma.
The authors conducted a review of the literature of cost-effectiveness analyses regarding management of Barrett's esophagus, including screening, surveillance, and treatment strategies. Because of the presence of multiple systematic reviews on this topic, they chose to focus on more recent economic analyses, with an emphasis on comparative modeling because these analyses have been demonstrated to achieve greater validity and impact when there are multiple competing strategies that are clinically reasonable to pursue. The authors identified areas of consensus across studies regarding management strategies and also areas that require additional empirical data.


Effects of argon plasma treatment on the osteoconductivity of bone grafting materials.
The osteoconductive properties of bone grafting materials represent one area of research for the management of bony defects found in the fields of periodontology and oral surgery. From a physico-chemical aspect, the wettability of the graft has been demonstrated to be one of the most important factors for new bone formation. It is also well-known that argon plasma treatment (PAT) and ultraviolet irradiation (UV) may increase the surface wettability and, consequently, improve the regenerative potential of the bone grafts. Therefore, the aim of the present in vitro study was to evaluate the effect of PAT and UV treatment on the osteoconductive potential of various bone grafts. The following four frequently used bone grafts were selected for this study: synthetic hydroxyapatite (Mg-HA), biphasic calcium phosphate (BCP), cancellous and cortical xenogenic bone matrices (CaBM, CoBM). Sixty-six serially numbered disks 10 mm in diameter were used for each graft material and randomly assigned to the following three groups: test 1 (PAT), test 2 (UV), and control (no treatment). Six samples underwent topographic analysis using SEM pre- and post-treatments to evaluate changes in surface topography/characteristics. Additionally, cell adhesion and cell proliferation were evaluated at 2 and 72 h respectively following incubation in a three-dimensional culture system utilizing a bioreactor. Furthermore, the effects of PAT and UV on immune cells were assessed by measuring the viability of human macrophages at 24 h. The topographic analysis showed different initial morphologies of the commercial biomaterials (e.g., Mg-HA and BCP showed flat morphology; BM samples were extremely porous with high roughness). The surface analysis following experimental treatments did not demonstrate topographical difference when compared with controls. Investigation of cells demonstrated that PAT treatment significantly increased cell adhesion of all 4 evaluated bone substitutes, whereas UV failed to show any statistically significant differences. The viability test revealed no differences in terms of macrophage adhesion on any of the tested surfaces. Within their limitations, the present results suggest that treatment of various bone grafting materials with PAT appears to enhance the osteoconductivity of bone substitutes in the early stage by improving osteoblast adhesion without concomitantly affecting macrophage viability. Treatment of bone grafts with PAT appears to result in faster osseointegration of the bone grafting materials and may thus favorably influence bone regeneration.


A significant association between intestinal helminth infection and anaemia burden in children in rural communities of Edo state, Nigeria.
Anaemia is estimated to affect half the school-age children and adolescents in developing countries. This study aimed to determine the prevalence of anaemia and evaluate the relationship of intestinal helminth infection on the anaemia status of children in the rural communities of Evbuomore, Isiohor, and Ekosodin. in the Ovia North East local government area of Edo State, Nigeria. Faecal samples and blood samples were obtained from 316 children aged 1-15 years. Faecal samples were examined using standard parasitological techniques, and anaemia was defined as blood haemoglobin <11 g/dL. Of the 316 children, 38.6% were anaemic: 75.9% of children in Evbuomore, 42.3% in Isiohor and 26.8% in Ekosodin. The overall parasite prevalence in the three communities were: Ascaris lumbricoides (75.6%), hookworm (16.19%) and Trichuris trichiura (7.3%). Malnutrition was patent; 37.0% of the children were stunted, 19.3% wasted, and 44.0% underweight. There was a statistically significant association between hookworm and Ascaris lumbricoides infection and anaemia (P < .001). Serum ferritin levels were more sensitive than haemoglobin in detecting anemia and were correlated with intestinal helminth infection. Intestinal helminth infection in a concomitant state of malnutrition is observed in this population. Intervention programmes should be aimed at control of intestinal helminth infection and iron supplementation.


A method for quantitation of apoplast hydration in Arabidopsis leaves reveals water-soaking activity of effectors of Pseudomonas syringae during biotrophy.
The plant apoplast has a crucial role in photosynthesis and respiration due to its vital function in gas exchange and transpiration. The apoplast is also a dynamic environment that participates in many ion and nutrient transport processes via plasma membrane-localized proteins. Furthermore, diverse microbes colonize the plant apoplast, including the hemibiotrophic bacterial pathogen, Pseudomonas syringae pv. tomato (Pto) strain DC3000. Pto DC3000 initiates pathogenesis upon moving through stomata into the apoplast and then proliferating to high levels. Here we developed a centrifugation-based method to isolate and quantify the apoplast fluid in Arabidopsis leaves, without significantly damaging the tissue. We applied the simple apoplast extraction method to demonstrate that the Pto DC3000 type III bacterial effectors AvrE1 and HopM1 induce hydration of the Arabidopsis apoplast in advance of macroscopic water-soaking, disruption of host cell integrity, and disease progression. Finally, we demonstrate the utility of the apoplast extraction method for isolation of bacteria proliferating in the apoplast.


Motor Cerebellar Connectivity and Future Development of Freezing of Gait in De Novo Parkinson's Disease.
To investigate the role of motor cerebellar connectivity in future development of freezing of gait, because it is a complex network disorder in Parkinson's disease (PD). We recruited 26 de novo patients with PD who experienced freezing of gait within 5 years from magnetic resonance imaging acquisition (vulnerable PD group), 61 patients with PD who had not experienced freezing of gait within 5 years (resistant PD group), and 27 healthy control subjects. We compared the resting state functional connectivity between the motor cerebellum and the whole brain between the groups. In addition, we evaluated the relationship between motor cerebellar connectivity and freezing of gait latency. The vulnerable PD group had increased functional connectivity between the motor cerebellum and parieto-occipito-temporal association cortices compared with the control group or the resistant PD group. Connectivity between lobule VI and the right superior parietal lobule, right fusiform gyrus, and left inferior temporal gyrus; between lobule VIIb and the right superior parietal lobule, right hippocampus, and right middle temporal gyrus; and between lobule VIIIb and the bilateral fusiform gyri, right middle occipital gyrus, and bilateral parietal lobes was inversely proportional to freezing of gait latency. The freezing of gait latency-related cortical functional connectivity from the motor cerebellum was also significantly higher in the vulnerable PD group compared with the control group, as well as the resistant PD group. The data suggest that the motor cerebellar functional connectivity with the posterior cortical areas play an important role in future development of freezing of gait in PD. © 2020 International Parkinson and Movement Disorder Society.


Effects of sit-to-stand training combined with transcutaneous electrical stimulation on spasticity, muscle strength and balance ability in patients with stroke: A randomized controlled study.
Sit-to-stand is a fundamental movement of human being for performing mobility and independent activity. However, Stroke people symptoms experience difficulty in conducting the sit-to-stand due to paralysis and especially ankle spasticity. Recently, transcutaneouselectrical- stimulation (TENS) is used to reduce pain but also to manage spasticity. The purpose of this study was to determine (1) whether TENS would lead to ankle spasticity reduction and (2) whether sit-to-stand training combined with TENS would improve spasticity, muscle strength and balance ability in stroke patients. Forty-stroke patients were recruited and were randomly divided into two groups: TENS group (n=20) and sham group (n=20). All participants underwent 30-sessions of sit-to-stand training (for 15-min, five-times per week for 6-weeks). Prior to each training session, 30-min of TENS over the peroneal nerve was given in TENS group, whereas sham group received non-electrically stimulated TENS for the same amount of time. Composite-Spasticity-Score was used to assess spasticity level of ankle plantar-flexors. Isometric strength in the extensor of hip, knee and ankle were measured by handheld dynamometer. Postural-sway distance was measured using a force platform. The spasticity score in the TENS group (2.6±0.8) improved significantly greater than the sham group (0.7±0.8, p<0.05). The muscle strength of hip extensor in the TENS group (2.7±1.1kg) was significantly higher than the sham group (1.0±0.8kg, p<0.05). Significant improvement in postural-sway was observed in the TENS group compared to the sham group (p<0.05). Thus, sit-to-stand training combined with TENS may be used to improve the spasticity, balance function and muscle strength in stroke patients.


A phylogenetic study of sulphation factor activity in 26 species.
Somatomedin (sulphation factor) activity was measured in 26 species by a porcine cartilage bioassay. Although non-dialysable inhibitory factors were present in many species, sulphation factor activity was present in the sera of all the vertebrates studied but was absent from the invertebrates.


PchR-box recognition by the AraC-type regulator PchR of Pseudomonas aeruginosa requires the siderophore pyochelin as an effector.
Under iron limitation, the opportunistic human pathogen Pseudomonas aeruginosa produces the siderophore pyochelin. When secreted into the extracellular environment, pyochelin complexes ferric ions and delivers them, via the outer membrane receptor FptA, to the bacterial cytoplasm. Extracellular pyochelin also acts as a signalling molecule, inducing the expression of pyochelin biosynthesis and uptake genes by a mechanism involving the AraC-type regulator PchR. We have identified a 32 bp conserved sequence element (PchR-box) in promoter regions of pyochelin-controlled genes and we show that the PchR-box in the pchR-pchDCBA intergenic region is essential for the induction of the pyochelin biosynthetic operon pchDCBA and the repression of the divergently transcribed pchR gene. PchR was purified as a fusion with maltose-binding protein (MBP). Mobility shift assays demonstrated specific binding of MBP-PchR to the PchR-box in the presence, but not in the absence of pyochelin and iron. PchR-box mutations that interfered with pyochelin-dependent regulation in vivo, also affected pyochelin-dependent PchR-box recognition in vitro. We conclude that pyochelin, probably in its iron-loaded state, is the intracellular effector required for PchR-mediated regulation. The fact that extracellular pyochelin triggers this regulation suggests that the siderophore can enter the cytoplasm.


The relationship between biomechanical variables and driving performance during the golf swing.
Swing kinematic and ground reaction force data from 308 golfers were analysed to identify the variables important to driving ball velocity. Regression models were applied at four selected events in the swing. The models accounted for 44-74% of variance in ball velocity. Based on the regression analyses, upper torso-pelvis separation (the X-Factor), delayed release (i.e. the initiation of movement) of the arms and wrists, trunk forward and lateral tilting, and weight-shifting during the swing were significantly related to ball velocity. Our results also verify several general coaching ideas that were considered important to increased ball velocity. The results of this study may serve as both skill and strength training guidelines for golfers.


Organic pollutant degradation by UV/peroxydisulfate process: Impacts of UV light source and phosphate buffer.
In recent years, ultraviolet (UV) based advanced oxidation processes have been extensively studied for degradation of refractory organic pollutants in water and wastewater, and selection of an appropriate UV light source is an important issue. In this study, bench-scale tests were conducted on a mini-fluidic photoreaction system (MFPS) to determine the degradation kinetics of methylene blue (MB) by UV/peroxydisulfate (UV/PDS) process equipped with a low-pressure UV (LPUV), vacuum UV (VUV)/LPUV, or medium-pressure UV (MPUV) mercury vapor lamp. Results indicate that MB degradation by UV/PDS with various light sources all followed the pseudo-first order kinetics, and the photon fluence-based rate constant (kp,λ') had a descending order of: VUV/LPUV/PDS ≫ MPUV/PDS > LPUV/PDS. Moreover, it is noted that phosphate buffer (PB) notably inhibited MB degradation: the kp,LPUV', kp,VUV/LPUV' and kp,MPUV' decreased by 35.0%, 44.9% and 37.5% with the PB concentration increasing from 0 to 20 mM, respectively. The maximal decrease in kp,VUV/LPUV' was ascribed to a strong competition of PB for VUV photons. Thereafter, pilot-scale tests were conducted to evaluate the practical performance of UV/PDS in terms of the electrical energy consumption per order (EEO). It was found again that the VUV/LPUV lamp was the optimal light source in UV/PDS for organic pollutant degradation. This study helps optimize the UV/PDS process for its practical application to water and wastewater treatment.


Removal of phenyl-urea herbicides in ultrapure water by ultrafiltration and nanofiltration processes.
Membrane filtration of four phenyl-urea herbicides (linuron, diuron, chlortoluron, and isoproturon) dissolved in ultrapure water was studied in a laboratory cross-flow device in batch concentration mode (with recycling of the retentate stream). Three UF (MWCO of 20 000, 5000 and 2000Da) and three NF (MWCO of 150-300Da) membranes were used. The influence of the main operating conditions (transmembrane pressure, tangential velocity, temperature, pH, and MWCO of the membranes) on the steady-state permeate fluxes and the retention factors of the phenyl-ureas was evaluated. The herbicide mass adsorbed onto the membranes was also determined, and the contribution of the fouling resistance to the total resistance to permeate flux was much lower than the inherent resistance of the clean membranes.


Improved fusion technique. III. The growth promoting activity of lipopolysaccharide, dextran sulfate, and red cell lysate added to Hy-clone calf serum.
A new high quality young-calf serum, Hy-clone calf serum (HcCS), was tested for use in hybridoma culture. This calf serum alone had little growth promoting activity and was much inferior to fetal calf serum (FCS). Red cell lysate (RCL) used in combination with the young-calf serum showed very good growth promoting activity. Growth was increased about threefold over that in the presence of FCS. However, HcCS and RCL could not substitute for feeder cells when hybridomas were cultured as single cells under conditions of limiting dilution. It is thought likely that the potent growth promoting factor in red cell lysate is hemoglobin.


Upregulation of cell surface estrogen receptor alpha is associated with the mitogen-activated protein kinase/extracellular signal-regulated kinase activity and promotes autophagy maturation.
Recently, accumulating evidence has implicated the dysregulation of autophagy as underlying the pathophysiology of several neurodegenerative diseases. The human neuronal cell line SH-SY5Y was exposed to 1-Methyl-4-phenylpyridinium (MPP(+)). The mechanism is that the sustained activation of the MAPK/ERK pathway by MPP(+) alters autophagy selectively at the maturation step, significant increasing in autophagy formation and delaying in autophagy degradation in SHSY5Y cells. In this study, we provided evidences that estrogen was capable of promoting SHSY5Y cells survival in MPP(+)-treated group. In particular, the up-regulation of mERα, but not mERβ, was associated with a rapid and transient activation of ERK phosphorylation compatible with promoting autophagy maturation. The up-regulation of mERα changed the sustained activation of ERK phosphorylation in MPP(+)-treated group into a temporary activation. Taken together, these findings strongly support that the expression of mERα promotes the maturation of autophagosomes into functional autolysosomes by regulating ERK, determining SHSY5Y cells survival.


Osteochondral Autograft: Proceedings of the International Consensus Meeting on Cartilage Repair of the Ankle.
Treatment guidelines for cartilage lesions of the talus have been based on both low quality and low levels of evidence. Therefore, an international consensus group of experts was convened to collaboratively advance toward consensus opinions on key topics regarding cartilage lesions of the talus. The purpose of this consensus article is to explain the process and delineate the consensus statements derived from this consensus meeting on the use of "osteochondral autograft" for osteochondral lesions of the talus. Seventy-five international experts in cartilage repair of the ankle representing 25 countries and 1 territory were convened and participated in a process based on the Delphi method of achieving consensus. Questions and statements were drafted within 11 working groups focusing on specific topics within cartilage repair of the ankle, after which a comprehensive literature review was performed and the available evidence for each statement was graded. Discussion and debate occurred in cases where statements were not agreed upon in unanimous fashion within the working groups. A final vote was then held, and the strength of consensus was characterized as follows: consensus, 51% to 74%; strong consensus, 75% to 99%; and unanimous, 100%. A total of 14 statements on osteochondral autograft reached consensus during the 2017 International Consensus Meeting on Cartilage Repair of the Ankle. Two achieved unanimous support, 11 reached strong consensus (greater than 75% agreement), and 1 achieved consensus. All statements reached at least 67% agreement. This international consensus derived from leaders in the field will assist clinicians with osteochondral autograft as a treatment strategy for osteochondral lesions of the talus.


The effect of insufflation pressure on CO(2) pneumoperitoneum and embolism in piglets.
We conducted this study to investigate the effect of insufflation pressure on the pathophysiology of CO(2) pneumoperitoneum and embolism in an infant model. Twenty anesthetized piglets had stepwise intraperitoneal insufflation with CO(2) for 15 min at pressures ranging from 5 to 20 mm Hg. The piglets were ventilated to baseline normocarbia (ETCO(2) = 30 mm Hg, PaCO(2) = 38 mm Hg) before beginning each insufflation. CO(2) was then insufflated IV in 15 of these piglets at the same pressures. There was no reduction of blood pressure or cardiac output with intraperitoneal insufflation, but the stroke volume declined significantly (*P < 0.05) from (mean +/- SE) 10.6 +/- 1.3 mL to 8.5 +/- 1.3* mL and from 10.0 +/- 1.4 mL to 7.2 +/- 1.2* mL at 15 and 20 mm Hg insufflation pressure, respectively. Abdominal insufflation at 5, 10, 15, and 20 mm Hg caused an increase in ETCO(2) to 31.7 +/- 0.8 mm Hg, 35.6 +/- 1.2* mm Hg, 37.5 +/- 1.5* mm Hg, and 40.1 +/- 1.8* mm Hg and in PaCO(2) to 41.1 +/- 1.3* mm Hg, 44.2 +/- 1.4* mm Hg, 49.9 +/- 1.8* mm Hg, and 53.0 +/- 2.1* mm Hg, respectively. In contrast, the ETCO(2)decreased to 19.4 +/- 1.5* mm Hg, 20.4 +/- 1.4 mm Hg, 15.2 +/- 2.1* mm Hg, and 10.6 +/- 2.0* mm Hg with IV insufflation using the same pressures. IV insufflation caused marked hypotension and mortality. As the insufflation pressure increased, the mortality increased (0 in 15, 1 in 15, 1 in 14, and 6 in 13* at 5, 10, 15, and 20 mm Hg; *P < 0.05 vs 0 in 15, 1 in 15, and 1 in 14). This study suggests that although intraperitoneal insufflation up to 20 mm Hg may be tolerated hemodynamically, the lowest possible pressure should be used to reduce hypercarbia. A low insufflation pressure may also prevent mortality from CO(2) embolism. The lowest pressure possible should be used when inflating the abdomen with CO(2) to perform a laparoscopy in babies. A low pressure allows better ventilation and may prevent mortality if CO(2) is accidentally injected into a vein.


Agonist-occupied A3 adenosine receptors exist within heterogeneous complexes in membrane microdomains of individual living cells.
G protein-coupled receptors are known to be organized within different membrane compartments or microdomains of individual cells. Here, we have used a fluorescent A3 adenosine receptor (A3-AR) agonist, ABEA-X-BY630, and the technique of fluorescence correlation spectroscopy (FCS) to investigate the diffusional characteristics of functional agonist-occupied A3-AR complexes in single living cells. In Chinese hamster ovary cells expressing the human A3-AR, the fluorescent A3-AR agonist was able to inhibit forskolin-stimulated [3H]cAMP production (pEC50=8.57), and this was antagonized by the A3-selective antagonist MRS1220 (pK(B)=9.32). The fluorescent ligand also stimulated phosphoinositide hydrolysis (pEC50=7.34). Ligand binding to the A3-AR on the membranes of single cells and subsequent increases in single cell [Ca2+]i were monitored simultaneously in real time using confocal microscopy. FCS measurements in small-membrane microdomains (approximately 0.2 microm2) revealed two agonist-occupied A3-AR components with differing diffusion characteristics (diffusion coefficients=2.65x10(-8) and 1.19x10(-9) cm2/s, respectively). The binding of ligand to these two components was reduced from 5.1 and 14.9 to 2.6 and 3.3 receptors/microm2, respectively, by MRS1220 (100 nM). These data provide direct evidence for at least two populations of agonist-occupied A3-receptor complexes, showing different motilities within the membrane of single living cells.


Near-infrared spectrometric determination of dipyrone in closed ampoules.
The present paper proposes an analytical method for fast near-infrared (NIR) determination of dipyrone in injectable formulations with a nominal content of 50.0%mv(-1) without violation of the ampoule. For this purpose, two multivariate calibration methods are evaluated, namely Partial-Least-Squares (PLS) and Multiple Linear Regression (MLR) with variable selection by the Successive Projections Algorithm (SPA). The resulting models comprised four latent variables (PLS) and five spectral variables (MLR-SPA). Appropriate predictions were obtained in both cases, with RMSEP values of 0.39 (PLS) and 0.35%mv(-1) (MLR-SPA) and correlation coefficients of 0.9970 (PLS) and 0.9975 (MLR-SPA) for a calibration range of 40-60%mv(-1). No systematic error was observed and no significant differences were found between the predicted and reference values, according to a paired t-test at 95% confidence level.


Author Correction: Review of the genus Brachytarsophrys (Anura: Megophryidae), with revalidation of Brachytarsophrys platyparietus and description of a new species from China.
Following the publication of our paper (Li et al., 2020), it has come to our attention that, due to our oversight, the contributions of some colleagues were not reflected in the "ACKNOWLEDGEMENTS" section. Hereby, we wish to add the following to the "ACKNOWLEDGEMENTS" section as a correction.


No compelling positive association between ovarian hormones and wearing red clothing when using multinomial analyses.
Several studies report that wearing red clothing enhances women's attractiveness and signals sexual proceptivity to men. The associated hypothesis that women will choose to wear red clothing when fertility is highest, however, has received mixed support from empirical studies. One possible cause of these mixed findings may be methodological. The current study aimed to replicate recent findings suggesting a positive association between hormonal profiles associated with high fertility (high estradiol to progesterone ratios) and the likelihood of wearing red. We compared the effect of the estradiol to progesterone ratio on the probability of wearing: red versus non-red (binary logistic regression); red versus neutral, black, blue, green, orange, multi-color, and gray (multinomial logistic regression); and each of these same colors in separate binary models (e.g., green versus non-green). Red versus non-red analyses showed a positive trend between a high estradiol to progesterone ratio and wearing red, but the effect only arose for younger women and was not robust across samples. We found no compelling evidence for ovarian hormones increasing the probability of wearing red in the other analyses. However, we did find that the probability of wearing neutral was positively associated with the estradiol to progesterone ratio, though the effect did not reach conventional levels of statistical significance. Findings suggest that although ovarian hormones may affect younger women's preference for red clothing under some conditions, the effect is not robust when differentiating amongst other colors of clothing. In addition, the effect of ovarian hormones on clothing color preference may not be specific to the color red.


Return to sports following lower limb musculoskeletal tumor surgery-A systematic review.
Survivors of primary malignant musculoskeletal tumors often face long-term disability. Clinicians at present are unable to provide evidence-based advice about returning to sports, which is important for active patients. Identify patients returning to sports. Describe the sporting activities in which patients participate. Identify the outcome measures used to assess return to sport. Identify barriers preventing return to sports. Systematic review. A comprehensive search strategy was used to identify relevant studies combining the following concepts: (1) Bone/Soft tissue tumor, (2) Lower limb, (3) Surgical interventions, and (4) Sports. Studies were selected according to eligibility criteria with the consensus of three authors (MTB, FS, and CG). Twenty-two studies were selected, published between 1985 and 2020, including 1005 patients. Fifteen of the 22 studies had valid data on return to sports, with 705 participants, of which 412 (58.4%) returned to some form of sport such as swimming and cycling, at a mean follow-up period of 7.6 years. Four studies directly compared limb sparing surgery and amputation; none of these were able to identify a difference in sports participation or ability. There is insufficient published research to provide guidance for patients with respect to return to sports following musculoskeletal tumors. Future prospective studies are needed to collect better pre- and post-treatment data at multiple time intervals. Validated clinical and patient sports participation outcomes such as type of sports, level, frequency, and validated sports-specific outcome scores should be recorded. In particular, more comparison between limb sparing surgery and amputation would be welcome.


Subclinical Sjögren's syndrome: a significant 67gallium accumulation in the orbits and parotid glands.
An 8-year-old girl with hypergammaglobulinemia showed an abnormal 67gallium accumulation in the orbits and parotid glands. Although she did not have any subjective siccant complaints, reported typical histopathological and sialographic changes suggesting Sjögren's syndrome (SjS) were observed in the salivary glands. Gallium scintigram might be a valuable and non-invasive diagnostic tool in the diagnosis of children with SjS without sicca symptoms.


Dual-mode behaviour in the human accommodation system.
The human accommodation system was investigated using a dynamic infra-red optometer to determine whether its feedback control process exhibited dual-mode behaviour. Ramp changes of target accommodative demand presented monocularly at various velocities (0.5-0.6 D s-1) with a fixed 2 D amplitude elicited two modes of behaviour. Smooth ramp movements were observed for low-velocity stimuli, whereas step responses predominated for high-velocity stimuli. At intermediate velocities, combined step-ramp responses were more prevalent. The characteristic of these step-ramp responses showed two important properties that suggest a pre-programming mechanism. The amplitude of the step portion of a step-ramp response approximated the value of the ramp stimulus at the time corresponding to the end of the step movement. Also, the initial step amplitude increased with increasing target ramp velocity. It appears that the amplitude of the preprogrammed accommodative movement was based on an estimate of the expected ramp target position at the end of the response. These findings are consistent with a recent unifying theory of oculomotor control in which the various oculomotor subsystems (versional and vergence eye movements and accommodation) each has fast and slow components. The fast component exhibits preprogramming, while the slow component shows continuous feedback control.


Life-cycle analysis of bio-based aviation fuels.
Well-to-wake (WTWa) analysis of bio-based aviation fuels, including hydroprocessed renewable jet (HRJ) from various oil seeds, Fischer-Tropsch jet (FTJ) from corn-stover and co-feeding of coal and corn-stover, and pyrolysis jet from corn stover, is conducted and compared with petroleum jet. WTWa GHG emission reductions relative to petroleum jet can be 41-63% for HRJ, 68-76% for pyrolysis jet and 89% for FTJ from corn stover. The HRJ production stage dominates WTWa GHG emissions from HRJ pathways. The differences in GHG emissions from HRJ production stage among considered feedstocks are much smaller than those from fertilizer use and N2O emissions related to feedstock collection stage. Sensitivity analyses on FTJ production from coal and corn-stover are also conducted, showing the importance of biomass share in the feedstock, carbon capture and sequestration options, and overall efficiency. For both HRJ and FTJ, co-product handling methods have significant impacts on WTWa results.


Health-related quality of life, one-year costs and economic evaluation in extracorporeal membrane oxygenation in critically ill adults.
This study reports on survival and health related quality of life (HRQOL) after extracorporeal membrane oxygenation (ECMO) treatment and the associated costs in the first year. Prospective observational cohort study patients receiving ECMO in the intensive care unit during August 2017 and July 2019. We analyzed all healthcare costs in the first year after index admission. Follow-up included a HRQOL analysis using the EQ-5D-5L at 6 and 12 months. The study enrolled 428 patients with an ECMO run during their critical care admission. The one-year mortality was 50%. Follow up was available for 124 patients at 12 months. Survivors reported a favorable mean HRQOL (utility) of 0.71 (scale 0-1) at 12 months of 0.77. The overall health status (VAS, scale 0-100) was reported as 73.6 at 12 months. Mean total costs during the first year were $204,513 ± 211,590 with hospital costs as the major factor contributing to the total costs. Follow up costs were $53,752 ± 65,051 and costs of absenteeism were $7317 ± 17,036. At one year after hospital admission requiring ECMO the health-related quality of life is favorable with substantial costs but considering the survival might be acceptable. However, our results are limited by loss of follow up. So it may be possible that only the best-recovered patients returned their questionnaires. This potential bias might lead to higher costs and worse HRQOL in a real-life scenario.


Pharmacognostical Studies.
The chapter deals with tools and techniques employed in pharmacognosy. Pharmacognostic evaluation helps to screen the commercial varieties, substitutes, adulterants and any other quality control of the drugs. It is a simple and reliable tool, helps to obtain information about biochemical and physical properties of crude drug. Methods such as macroscopic and microscopic analysis, maceration, histochemical colour reaction, photomicrography, organoleptic character of plant powder and extracts, fluorescence analysis of plant powder with different chemical reagents, determination of pH of plant powder, water solubility index (WSI) and water absorption index (WAI) and acid value are discussed.


Changes of cytosolic Ca(2+) fluorescence intensity and plasma membrane calcium channels of maize root tip cells under osmotic stress.
The changes of cytosolic Ca(2+) fluorescence intensity and the activities of calcium channel of primary maize root tip cells induced by PEG6000 or abscisic acid (ABA) were studied by both confocal techniques and the whole-cell patch clamping in this study. The Ca(2+) fluorescence intensity increased while treated with PEG or ABA within 10 min, illuminating that Ca(2+) participated in the process of ABA signal transduction. For further proving the mechanism and origin of cytosolic Ca(2+) increase induced by PEG treatments, N,N,N',N'-tetraacetic acid (EGTA), Verapamil (VP) and Trifluoperazine (TFP) were added to the PEG solution in the experiments separately. The results showed that Ca(2+) fluorescence intensity induced by PEG was suppressed by both EGTA and VP obviously in the root tip cells. The Ca(2+) fluorescence intensity of plants changed after the addition of CaM inhibitor TFP while subjected to osmotic stress, which seemed to show that CaM participated in the process of signal transduction of osmotic stress too. The mechanism about it is unknown today. Further, a hyperpolarization-activated calcium permeable channel was recorded in plasma membrane of maize root tip cells. The Ca(2+) current (I(Ca)) intensity increased remarkably after PEG treatment, and the open voltage of the calcium conductance increased. Similar changes could be observed after ABA treatment, but the channel opened earlier and the current intensity was stronger than that of PEG treatment. The activation of calcium channel initiated by PEG strongly was inhibited by EGTA, VP or TFP respectively. The results revealed that Ca(2+) participated in the signals transduction process of osmotic stress, and the cytosolic free Ca(2+) increase by osmotic stress mainly came from the extracellular, and some came from the release of cytoplasmic calcium pool.


Performance of the ACMG-AMP criteria in a large familial renal glucosuria cohort with identified SLC5A2 sequence variants.
Familial Renal Glucosuria (FRG) is a co-dominantly inherited trait characterized by orthoglycaemic glucosuria. From 2003 to 2015 we have reported several cohorts validating SLC5A2 (16p11.2), encoding SGLT2 (Na+/glucose cotransporter family member 2), as the gene responsible for FRG. The aim of this work was to validate the variants identified in our extended FRG cohort of published, as well more recent unreported cases, according to the ACMG-AMP 2015 criteria. Forty-six variants were evaluated, including 16 novel alleles first described in this study. All are rare, ultra-rare or absent from population databases and most are missense changes. According to the ACMG-AMP standards, only 74% of the variants were classified as P/LP. The lack of descriptions of unrelated patients with similar variants or failing to test additional affected family members, averted a conclusion for pathogenicity in the alleles that scored VUS, highlighting the importance of both family testing and variant reporting. Finally, the cryo-EM structure of the hSGLT2-MAP17 complex in the empagliflozin-bound state improved the ACMG-AMP pathogenicity score by identifying critical/functional protein domains.


Absenteeism Due to Mental Disorders in Agents Fighting Endemic Diseases in Ceará/Northeast Brazil.
The purpose of this study is to quantify the days away from work by mental disorders (MDs) in endemic fighting agents (EFAs) in the state in Northeast Brazil. This is a historical cohort carried out from the survey of 584 expert medical records of EFA. The association measures were performed using Poisson regression models. The days away from work increased over time. Endemic fighting agents who have been on sick leaves by mood disorders ([cumulative incidence like relative risk (IRR)], 21.37; 95% CI, 12.36-36.93), alcoholism (IRR, 17.12; 95% CI, 10.04-29.34), and stress-related disorders (IRR, 10.32; 95% CI, 6.05-17.63) have a higher risk of absenteeism longer than 15 days. This study reinforces the importance of actions for coping with MD in EFA.


Interactions of attention-deficit/hyperactivity disorder therapeutic agents with the efflux transporter P-glycoprotein.
The objective of this study was to assess the potential interactions of the drug transporter P-glycoprotein with attention-deficit/hyperactivity disorder (ADHD) therapeutic agents atomoxetine--and the individual isomers of methylphenidate, amphetamine, and modafinil utilizing established in vitro assay. An initial ATPase assay indicated that both d- and l-methylphenidate have weak affinity for P-glycoprotein. The intracellular accumulation of P-glycoprotein substrates doxorubicin and rhodamine123 in the P-glycoprotein overexpressing cell line LLC-PK1/MDR1 was determined to evaluate potential inhibitory effects on P-glycoprotein. The results demonstrated that all compounds, except both modafinil isomers, significantly increased doxorubicin and rhodamine123 accumulation in LLC-PK1/MDR1 cells at higher concentrations. To investigate the P-glycoprotein substrate properties, the intracellular concentrations of the tested compounds in LLC-PK1/MDR1 and P-glycoprotein negative LLC-PK1 cells were measured in the presence and absence of the P-glycoprotein inhibitor PSC833. The results indicate that the accumulation of d-methylphenidate in LLC-PK1 cells was 32.0% higher than in LLC-PK1/MDR1 cells. Additionally, coadministration of PSC833 leads to 52.9% and 45.6% increases in d-modafinil and l-modafinil accumulation, respectively, in LLC-PK1/MDR1 cells. Further studies demonstrated that l-modafinil transport across LLC-PK1/MDR1 cell monolayers in the basolateral-to-apical (B-A) direction was significantly higher than in the apical-to-basolateral (A-B) direction. PSC833 treatment significantly decreased the transport of l-modafinil in B-A direction. In conclusion, our results suggest that all tested agents with the exception of modafinil isomers are relatively weak P-glycoprotein inhibitors. Furthermore, P-glycoprotein may play a minor role in the transport of d-methylphenidate, d-modafinil, and l-modafinil.


Three-dimensional transmission electron microscopy and its application to mitosis research.
Transmission electron microscopy produces images that are projections of the original object, with the consequence that features from different depths of the specimen overlap and give a confusing image. This problem is overcome by reconstructing the object in 3D from a series of 2D views using either serial thin section reconstruction or electron tomography. In the serial section approach, the series of 2D views is generated from images of successive serial sections cut thin enough to be effectively 2D slices of the specimen. For electron tomography the series of 2D views is generated by tilting a single, usually thicker, section in the electron beam. Resolution in the depth dimension is limited to twice the section thickness for serial section reconstruction and is determined by the number of tilt views collected (i.e., by the fineness of the angular interval between successive tilt views) for electron tomography. Both methods produce distorted 3D reconstructions because of missing material and alignment difficulties in the case of serial sections and the limited angular tilt range in the case of electron tomography. However, techniques have evolved for minimizing and circumventing these distortions and, as long as the user is aware of the limitations, misinterpretations can be avoided. Since electron tomography provides better resolution (generally 5-20 nm), it is the method of choice for determining detailed structural interactions such as the depth of kinetochore MT penetration into the kinetochore outer plate. On the other hand, serial section reconstruction is more effective for projects that require tracking through a complete object in the specimen, such as counting the number of kinetochore MTs on each kinetochore. If the project requires finding a relatively small object in a large specimen (e.g., finding centrioles in an oocyte), then it is sometimes advantageous to cut thicker plastic sections and analyze them via stereo viewing. The mitotic spindle, however, is generally too complex to be analyzed via stereo viewing. Currently, collapse of plastic sections in the electron beam limits the utility of serial section electron tomography. Once a 3D reconstruction is completed it must be analyzed with the 2D medium of the screen on a computer monitor. The easiest approach is usually to walk through the 3D reconstruction volume slice by slice. However, in order to appreciate 3D interactions, and to communicate the results to others, it is generally necessary to segment key components from the rest of the volume and use modeling and rendering techniques. Rendered surface views can easily be color coded and provided with a number of depth cues to simulate the surface viewing encountered in everyday life. In some instances, it is useful to look through a smaller portion of the reconstruction volume with "X-ray vision." This can accomplished by using volume rendering to create a series of semitransparent views from different tilt angles.


Insights on defeating coronavirus disease (COVID-19) outbreak and predicting tourist arrival on the Chinese Hainan Leisure Island during the COVID-19 pandemic.
Hainan province is a very popular leisure tourist arriving city in China. Coronavirus disease 2019 (COVID-19) emerged in China and rapidly in early 2020, and due to its rapid worldwide spread, the World Health Organization declared COVID-19 as a global emergency. During the COVID-19 pandemic in Hainan province, many businesses and economies were influenced in this unexpected event, especially in tourism. This study used 2 classical forecasting methods to predict the number of tourists on Hainan Leisure Island from September to December in the second half of 2020 and to summarize the COVID-19 fighting experience during the pandemic. In addition, the Hainan government implemented epidemic control measures to resume production and work, and promote new tourism measures to acquire superior COVID-19 protection. Winter's method provides a statistical model for predicting the number of visitors to Hainan under normal conditions. The trend analysis method considers the impact of the black swan event, an irregular event, and only uses the data under the influence of the event to predict according to the trend. If the impact of the black swan event (COVID-19) continues, the prediction can be made using this method. In addition, the Hainan government has undertaken timely and effective measures against COVID-19 to promote leisure tourism development.


Association of an anaplastic lymphoma kinase pathway signature with cell de-differentiation, neoadjuvant chemotherapy response, and recurrence risk in breast cancer.
Aberrant activation of anaplastic lymphoma kinase (ALK) signaling has been found to be involved in the tumorigenesis of multiple types of cancer. The aim of this study was to determine the role of this pathway in the pathogenesis of breast cancer. An ALK pathway signature that we generated previously was used to compute the ALK pathway activity in 6381 breast cancer samples from 42 microarray datasets, and the associations between ALK pathway signature score and clinical variables were examined using logistic regression and survival analyses. Our results indicated that high ALK pathway activity was a significant risk factor for hormone receptor-negative, high-grade breast cancer in the 42 datasets. ALK pathway activity was positively associated with pathological complete response (pCR) in 15 datasets annotated with patient's neoadjuvant chemotherapy response information (overall odds ratio = 1.67, P < 0.01), and this association was more significant in HER2-negative and grade 1&2 tumors than in HER2-positive and grade 3 tumors. ALK pathway activity was also positively associated with recurrence risk in breast cancer patients from 30 datasets annotated with survival information (overall hazard ratio = 1.21, P < 0.01), particularly in patients with age > 50 years old, with positive lymph nodes, or with residual disease after neoadjuvant chemotherapy. ALK may be involved in breast cancer tumorigenesis, and ALK pathway signature score may serve as a prognostic biomarker for breast cancer.


Dependence of Cu, Pb and Zn remobilization on physicochemical properties of marine sediments.
The resuspension of 65 marine sediments was simulated in the laboratory with elutriates from 30 different sites from the north coast of Spain. The partitioning of Cu, Pb and Zn between sediment and elutriate was studied as a function of different physicochemical characteristics of the sediment: organic matter (OM), fine fraction (FF), redox potential and acid volatile sulfides (AVS). Mean remobilization factors (RF) -calculated as metal concentration in the elutriate (μg/L) divided by metal concentration in the sediment (μg/g dry weight)- were 0.072 for Cu, 0.012 for Pb and 0.071 for Zn. Remobilization of Pb was significantly lower than that of Cu and Zn. Although AVS, OM and FF presented a strong intercorrelation, OM explained great part of the variability on Cu and Pb remobilization while AVS did it for Zn. A multiple regression model considering both OM and AVS explained slightly better the remobilization of Pb and Cu, but not that of Zn.


A multi-head residual connection GCN for EEG emotion recognition.
Electroencephalography (EEG) emotion recognition is a crucial aspect of human-computer interaction. However, conventional neural networks have limitations in extracting profound EEG emotional features. This paper introduces a novel multi-head residual graph convolutional neural network (MRGCN) model that incorporates complex brain networks and graph convolution networks. The decomposition of multi-band differential entropy (DE) features exposes the temporal intricacy of emotion-linked brain activity, and the combination of short and long-distance brain networks can explore complex topological characteristics. Moreover, the residual-based architecture not only enhances performance but also augments classification stability across subjects. The visualization of brain network connectivity offers a practical technique for investigating emotional regulation mechanisms. The MRGCN model exhibits average classification accuracies of 95.8% and 98.9% for the DEAP and SEED datasets, respectively, highlighting its excellent performance and robustness.


Sodium appetite and cerebrospinal fluid sodium concentration during hypovolemia.
A lowered concentration of sodium in the cerebrospinal fluid (CSF) has been suggested as a critical signal for the elicitation of sodium appetite. This was evaluated in naive male Long-Evans rats made hypovolemic by subcutaneous injection of the colloid polyethylene glycol (PEG) in a hyperoncotic concentration. At 2, 5 or 8 h after PEG or vehicle injection, under conditions of availability or non-availability of water and 0.3 M NaCl for ingestion, the rats were anesthetized and samples of CSF and blood were taken. Water intakes of the PEG treated rats were elevated at 5 and 8 h, and 0.3 M NaCl intakes were elevated at 8 h in comparison to vehicle treated rats. When fluids were not available, CSF sodium concentrations were greater in PEG than in vehicle treated rats at 2, 5 and 8 h. When fluids were available, CSF sodium concentrations were greater in PEG than in vehicle treated rats at 5 and at 8 h. There was no correlation between CSF sodium concentration and saline intake. In summary, a lowered CSF sodium concentration is not a necessary condition for sodium appetite in the hypovolemic rat.


Advanced glycation end products suppress lysyl oxidase and induce bone collagen degradation in a rat model of renal osteodystrophy.
Renal osteodystrophy (ROD) is a major problem in patients with renal insufficiency. The present study was designed to elucidate the role of bone collagen changes and osteoblast differentiation in a rat model of ROD pathogenesis induced by adenine. Typical characteristics of renal failure, including increased serum urea nitrogen, creatinine, inorganic phosphorus, and intact parathyroid hormone levels, and decreased serum calcium and 1,25(OH)2D3 levels, were observed in adenine-induced rats. Micro-computed tomography analysis of the femur in adenine-induced rats showed decreased bone mineral density and osteoporotic changes, confirmed by the three-point bending test. The cancellous bone histomorphometric parameters of the tibia showed increased osteoblast number, decreased osteoclast surface with peritrabecular fibrosis, and increased osteoid tissue, indicating a severe mineralization disorder similar to clinical ROD. Scanning and transmission electron microscopy revealed irregular alignment and increased diameter of bone collagen fibrils in adenine-induced rats. Protein expression analysis showed greater accumulation of advanced glycation end products (AGEs) in peritrabecular osteoblasts of adenine-induced rats than in the controls. In contrast, suppressed expression of runt-related transcription factor 2, alkaline phosphatase, secreted phosphoprotein 1 (Spp1), and lysyl oxidase (Lox) mRNA levels, particularly the amount of active LOX protein, were observed. In in-vitro experiments, mineralizing MC3T3-E1 osteoblastic cells stimulated with AGE-modified bovine serum albumin had attenuated the expression of Spp1 mRNA levels and active LOX protein, with a decrease in extracellular nodules of mineralization. These observations provide clues to ROD pathogenesis, as they indicate that the suppression of osteoblast differentiation and decreased active LOX protein associated with accumulation of AGEs in osteoblasts caused structural abnormalities of bone collagen fibrils and a severe mineralization disorder, leading to bone fragility.


Relative efficacy of two token economy procedures for decreasing the disruptive classroom behavior of retarded children.
Numerous studies have demonstrated that disruptive classroom behavior can be decreased by delivering tokens contingent upon periods of time during which children do not engage in it or by removing tokens contingent upon its occurrence. To date, the best controlled of these studies have consistently reported the two procedures to be qually effective. However, in these studies, token the two procedures to be equally effective. However, in these studies, token contingencies have been combined with instructions regarding the contigencies. The present study compared these two procedures when no instructions were given regarding the token contingencies. Token delivery was not effective in decreasing disruptive behavior in any of the children, while a combination of token delivery and removal was effective for three of four children. The results token delivery and removal was effective for three of four children. The results suggest that the combined procedure may be effective with certain populations that are not readily controlled by instructions.


Regulation of Argonaute slicer activity by guide RNA 3' end interactions with the N-terminal lobe.
Structural studies indicate that binding of both the guide RNA (siRNA and miRNA) and the target mRNA trigger substantial conformational changes in the Argonaute proteins. Here we explore the role of the N-terminal lobe (and its PAZ domain) in these conformational changes using biochemical and cell culture-based approaches. In vitro, whereas deletion (or mutation) of the N-terminal lobe of DmAgo1 and DmAgo2 had no effect on binding affinity to guide RNAs, we observed a loss of protection of the 3' end of the guide RNA and decreased target RNA binding; consistent with this, in cells, loss of function DmAgo1 PAZ variant proteins (PAZ6 and ΔN-PAZ) still bind RNA, although the RNAs are shorter than normal. We also find that deletion of the N-terminal lobe results in constitutive activation of endogenous PIWI domain-based cleavage activity in vitro, providing insights into how cleavage activity may be regulated in vivo in response to different types of pairing interactions with the target mRNAs.


Expression patterns of WT1 and PRAME in acute myeloid leukemia patients and their usefulness for monitoring minimal residual disease.
Both WT1 and PRAME are highly expressed in acute myeloid leukemia (AML) patients. To assess the efficacy of their simultaneous detection for the purpose of monitoring minimal residual disease (MRD), we used real-time quantitative RT-PCR to quantify both WT1 and PRAME transcript levels in the bone marrow of 204 newly diagnosed AML patients, and 21 patients were serially monitored for a median of 11 months. The 22 normal bone marrow samples which served as controls collectively expressed low levels of WT1 and PRAME. An increase of >1-log over the upper limit of normal bone marrow was defined as positive. The positive rates of WT1 and PRAME for all patients were 79.2% and 55.4%, respectively. For the 108 patients lacking a specific fusion gene, the positive rate of WT1 was significantly higher than that of PRAME (76.9% vs. 35.2%, P<0.001). PRAME was positive in 9/25 WT1 (-) patients and the log increase of PRAME was higher than that of WT1 in 12/83 WT1 (+) patients. Dynamic patterns of WT1 and PRAME during follow-up showed that a consistent elevation or a rise over time to exceed the normal range predicted clinical relapse. The exception was that one patient's WT1 significantly decreased at relapse compared to diagnosis. Therefore, a simultaneous detection of WT1 and PRAME might not only provide AML patients with either a positive or a more sensitive molecular marker for MRD monitoring. Moreover, it might also avoid false negativity in the case of expression alteration.


Feasibility of a machine vs free weight strength training program and its effects on physical performance in nursing home residents: a pilot study.
Resistance training holds promise for nursing home residents to prevent further disabilities, falls, and fractures. Free weight as well as machine training may offer an efficient option to improve physical performance, but the feasibility of these training regimes among elderly who require continuous institutional care is still open. (1) To examine the feasibility of a 3-month machine vs. free weight strength training program in institutionalized older adults, and (2) to determine the effects on physical performance. This study is a two-arm, single-blind, randomized controlled feasibility study within a nursing home. 45 institutionalized elderly men and women (aged 83.8 ± 8.0, 12 men, 33 women) were randomly divided into two groups. The two groups completed either a free weight (FWT) or machine training (MT) for 12 weeks, twice per week, 45-60 min per session, in an individually supervised format. Performance was assessed with the 11-step stair-climbing test, 10-m walk test, Timed Up and Go Test (TUG), 30-s Chair Rising Test (CRT), grip strength, body mass index. Indices of feasibility showed a recruitment and adherence rate of 53.6 and 87.5%, respectively. 35.6% of the participants dropped out after several weeks for personal reasons, illness, medical visits, or hospital stays. After the program no significant differences on motor performance were found between MT and FWT. However, there were significant improvements for both training groups on the TUG and the CRT. The present pilot study showed that it is feasible to conduct a strength training program in institutionalized participants. The more robust changes in motor function could serve as a basis for large randomized clinical trials.


In-Silico Screening of Lipid-Based Drug Delivery Systems.
This work proposes an in-silico screening method for identifying promising formulation candidates in complex lipid-based drug delivery systems (LBDDS). The approach is based on a minimum amount of experimental data for API solubilites in single excipients. Intermolecular interactions between APIs and excipients as well as between different excipients were accounted for by the Perturbed-Chain Statistical Associating Fluid Theory. The approach was applied to the in-silico screening of lipid-based formulations for ten model APIs (fenofibrate, ibuprofen, praziquantel, carbamazepine, cinnarizine, felodipine, naproxen, indomethacin, griseofulvin and glibenclamide) in mixtures of up to three out of nine excipients (tricaprylin, Capmul MCM, caprylic acid, Capryol™ 90, Lauroglycol™ FCC, Kolliphor TPGS, polyethylene glycol, carbitol and ethanol). For eight out of the ten investigated model APIs, the solubilities in the final formulations could be enhanced by up to 100 times compared to the solubility in pure tricaprylin. Fenofibrate, ibuprofen, praziquantel, carbamazepine are recommended as type I formulations, whereas cinnarizine and felodipine showed a distinctive solubility gain in type II formulations. Increased solubility was found for naproxen and indomethacin in type IIIb and type IV formulations. The solubility of griseofulvin and glibenclamide could be slightly enhanced in type IIIb formulations. The experimental validation agreed very well with the screening results. The API solubility individually depends on the choice of excipients. The proposed in-silico-screening approach allows formulators to quickly determine most-appropriate types of lipid-based formulations for a given API with low experimental effort. Graphical abstract.


Feeding native rumen microbial supplements increases energy-corrected milk production and feed efficiency by Holstein cows.
The objective of this study was to investigate the effects of 2 novel native microbial feed supplements (MFS) on milk yield, milk composition, dry matter intake (DMI), body weight (BW), body condition score (BCS), and efficiency. A total of 73 lactating Holstein cows were enrolled over 4 mo. During a 2-wk covariate period, baseline data were collected. Using a randomized complete block design, cows were blocked in groups of 3 by days in milk (DIM), production, and parity. Within blocks cows were randomly assigned to a basal diet with a 150 g/d ground corn topdress and no MFS (control), the basal diet supplemented with a 5 g/d dose of Clostridium beijerinckii ASCUSDY20 and Pichia kudriavzevii ASCUSDY21 in a carrier mixed with a 150 g/d ground corn topdress (MFS1), or the basal diet with a 5 g/d dose of Ruminococcus bovis ASCUSDY10, Butyrivibrio fibrisolvens ASCUSDY19, Clostridium beijerinckii ASCUSDY20, and Pichia kudriavzevii ASCUSDY21 in a carrier mixed in a 150 g/d ground corn topdress (MFS2) for 140 d. Cows were fed the diets as a total mixed ration (TMR) for ad libitum DMI once daily. Analysis showed numerical increases in performance variables such as milk yield and fat and protein contents in both MFS1 and MFS2, although the differences were not significant over time. For energy-corrected milk (ECM), other solids percentage, and BW there were significant treatment by time interactions. Cows fed MFS1 and MFS2 produced 1.4 kg/d and 1.6 kg/d more ECM than control cows, respectively (time-dependent treatment significance). Up to wk 13 of treatment, both MFS groups had higher ECM production than the control group but after this point MFS1 rapidly decreased to the same level as control. All cows displayed a gradual decrease in ECM after wk 13 but the MFS2 group remained greater than both control and MFS1. A similar trend in the content of other milk solids was observed. Cow BW was affected over time; both control and MFS2 cows gained weight at a similar rate throughout the study, whereas the weight gain of MFS1 cows decreased briefly from wk 13 to 17 before increasing again. Feed efficiency tended to be improved by MFS, with greater improvement by MFS2 than by MFS1. The DIM when the supplementation began had a significant impact on MFS2 efficacy, where the supplementation of native rumen microbes at an earlier DIM resulted in a greater gain in ECM over the 140-d trial. A similar trend was observed for MFS1, although the effect was not significant. Our results suggest that dairy cow lactation stage and energy requirements also play an important role in product efficacy.


Child with Wiskott-Aldrich syndrome underwent atypical immune reconstruction after umbilical cord blood transplantation: A case report.
Timely reconstitution of a donor-derived immune system is important for recovery and long-term survival of patients after allogeneic hematopoietic stem cell transplantation (HSCT). We describe a case of Wiskott-Aldrich syndrome (WAS) treated by umbilical cord blood transplantation (UCBT) with atypical immune reconstruction. A 1-year-old Chinese male infant was diagnosed with WAS. WAS gene sequencing identified the mutation c.777 + 1G>A (IVS8). On August 8, 2017, he was admitted to our hospital for HSCT. We selected an unrelated Human leukocyte antigen 6/10-matched donor for UCBT. After HSCT, the immune reconstitution process was atypical, the lymphocytes reached 0.5 × 109/L on day 23, and the neutrophils reached 0.5 × 109/L on day 34. The patient's recovery throughout the year was good. An increase in lymphocytes (especially T cells) earlier than granulocytes may be a marker of a good prognosis in UCBT.


Hypoglycemia in children.
The factors that sustain postabsorptive glucose concentrations have been analyzed and the adverse effects of various hypoglycemic disorders on these factors examined. The role of alanine has been reviewed and the importance of glycerol as a precursor of glucose and of ketones as a fuel substitute for glucose emphasized. Finally, we have suggested that fasting functional hypoglycemia replace ketotic hypoglycemia as a descriptive term and that we relinquish the concept of leucine-sensitive hypoglycemia as a specific entity.


Targeted lateral pelvic lymph node excision in the setting of total neoadjuvant therapy for rectal cancer.
Targeted lateral pelvic node dissection in the era of total neoadjuvant therapy.


Effects of Aging on Expression of Mic60 and OPA1 and Mitochondrial Morphology in Myocardium of Tibetan Sheep.
In order to investigate the effects of aging on the expression of Mic60 and OPA1 and mitochondrial morphology in plateau animals, the expression of Mic60 and OPA1 genes and proteins, and the morphology of mitochondria in the myocardium of adult and aged Tibetan sheep were investigated. The expression of Mic60 and OPA1 genes and OPA1 protein were higher (p < 0.05) in the myocardium of adult Tibetan sheep than in those of the aged ones. The number of mitochondrial cristae in the myocardium of adult was higher than that in aged (p < 0.05). The density of mitochondria in the myocardium of adult was higher than that in aged (p < 0.01). Compared with the adult Tibetan sheep, the mitochondrial crista of aged were relatively sparse, the crista membrane was wide, and the mitochondria were not closely linked, showing fragmentation. These results suggest that the myocardial mitochondria of the adult have better energy supply ability, indicating that aging can lead to the weakening of oxygen supply in the myocardial mitochondria of Tibetan sheep.


Colonizing the heart from the epicardial side.
The clinical use of stem cells, such as bone marrow-derived and, more recently, resident cardiac stem cells, offers great promise for treatment of myocardial infarction and heart failure. The epicardium-derived cells have also attracted attention for their angiogenic paracrine actions and ability to differentiate into cardiomyocytes and vascular cells when activated during cardiac injury. In a recent study, Chong and colleagues have described a distinct population of epicardium-derived mesenchymal stem cells that reside in a perivascular niche of the heart and have a broad multilineage potential. Exploring the therapeutic capacity of these cells will be an exciting future endeavor.


Effects of oxytocin-induced uterine hyperstimulation during labor on fetal oxygen status and fetal heart rate patterns.
The objective of the study was to evaluate effects of oxytocin-induced hyperstimulation on fetal oxygen saturation and fetal heart rate patterns. Uterine activity of 56 women was evaluated retrospectively for hyperstimulation lasting 30 minutes using 2 definitions: group 1: 5 or more but less than 6 contractions in 10 minutes (n = 102, 30-minute periods); group 2: 6 or more contractions in 10 minutes (n = 56, 30-minute periods). Fetal oxygen saturation and heart rate patterns during each period and the preceding 30 minutes of less than 5 contractions in 10 minutes were compared. Hyperstimulation was associated with significant oxygen desaturation: (group 1 = 10.68 [20%] decrease from 52.14 to 41.46; P < .001); group 2 = 15.34 [29%] decrease from 52.02 to 36.68: P < .001) and significantly more nonreassuring fetal heart rate characteristics, compared with normal uterine activity. Hyperstimulation is associated with negative effects on fetal status. The more contractions in 30 minutes, the more pronounced the effect.


Antioxidant and protective effect of latex of Calotropis procera against alloxan-induced diabetes in rats.
In the present study, dry latex (DL) of Calotropis procera possessing potent anti-inflammatory activity was evaluated for its antioxidant and anti-hyperglycemic effects against alloxan-induced diabetes in rats. Daily oral administration of DL at 100 and 400 mg/kg doses produced a dose-dependent decrease in the blood glucose and increase in the hepatic glycogen content. DL also prevented the loss of body weight in diabetic rats and brought down the daily water consumption to values comparable to normal rats. DL also produced an increase in the hepatic levels of the endogenous antioxidants, namely superoxide dismutase (SOD), catalase and glutathione, while it brought down the levels of thiobarbituric acid-reactive substances (TBARS) in alloxan-induced diabetic rats. The efficacy of DL as an antioxidant and as an anti-diabetic agent was comparable to the standard anti-diabetic drug, glibenclamide.


Reverse Locked-In Syndrome.
Basilar artery occlusion can cause locked-in syndrome, which is characterized by quadriplegia, anarthria, and limited communication via eye movements. Here, we describe an uncommon stroke syndrome associated with endovascular recanalization of the top of the basilar artery: "reverse locked-in syndrome." We report the case of a patient with atypical neurological deficits caused by acute ischemic stroke of the midbrain tegmentum. We perform neuroanatomic localization of the patient's infarcts by mapping the magnetic resonance imaging (MRI) data onto a brainstem atlas. A 61-year-old man presented with acute coma and quadriplegia due to top of the basilar artery occlusion. He underwent emergent endovascular thrombectomy, with successful recanalization of the basilar artery at 4 h and 43 min post-ictus. The patient regained consciousness and purposeful movement in all four extremities, but the post-procedure neurological examination demonstrated bilateral ptosis with complete pupillary and oculomotor paralysis. MRI revealed infarction of the bilateral oculomotor nuclei in the midbrain tegmentum. At 9-month follow-up, he had anisocoria and dysconjugate gaze, but was living at home and required minimal assistance in performing all activities of daily living. Since the patient's deficits were the exact opposite of those described in locked-in syndrome, we propose the term "reverse locked-in syndrome" to describe this neurological entity characterized by bilateral ptosis, non-reactive pupils, and ophthalmoplegia with preservation of consciousness and extremity motor function.


Reevaluating Emx gene phylogeny: homopolymeric amino acid tracts as a potential factor obscuring orthology signals in cyclostome genes.
Vertebrate Emx genes, retained as multiple copies, are expressed in a nested pattern in the early embryonic forebrain and required for its regionalization. This pattern seems to have originated in a vertebrate common ancestor; however, a previous analysis, reporting two lamprey Emx genes, claimed independent Emx gene duplications in both cyclostome (extant jawless fish) and gnathostome (jawed vertebrate) lineages after their divergence. This scenario is neither parsimonious nor consistent with the hypothesis that genome expansion occurred before the cyclostome-gnathostome split, which is supported by recent genome-wide analyses. We isolated and sequenced cDNA of two hagfish Emx genes and performed intensive molecular phylogenetic analyses, including the hagfish and/or lamprey Emx genes. The lamprey genes tended to attract each other in inferred phylogenetic trees, an effect that tended to be relaxed on inclusion of the hagfish genes. The results of these analyses suggest that cyclostome EmxB is orthologous to gnathostome Emx2, which was also supported by conserved synteny. Homopolymeric amino acid (HPAA) tracts represent a remarkable feature of the lamprey Emx sequences, and a comparative genome-wide scan revealed that lamprey proteins exhibit a unique pattern of HPAA tract accumulation. Our analysis, including hagfish Emx genes, suggests that gene duplications gave rise to Emx1, -2 and -3 before the cyclostome-gnathostome split. We propose that independent HPAA tract accumulations in multiple ancient duplicates, as identified in lamprey Emx gene products, may have led to erroneous identification of gene duplication in the lamprey lineage. Overall, our reanalysis favors the scenario that the nested Emx expression pattern in mouse and lamprey shares a common origin.


Intraoperative radiotherapy (IORT) versus whole-breast external beam radiotherapy (EBRT) in early stage breast cancer: results from SEER database.
Intraoperative radiotherapy (IORT) has emerged as an alternative to whole-breast external beam radiotherapy (EBRT) for early breast cancer. This study aimed to investigate the risk factors for survival after IORT or whole-breast EBRT in breast cancer patients. Breast cancer patients undergoing IORT or whole-breast EBRT were included from the surveillance, epidemiology, and end results database. Risk factors for overall survival (OS) and cancer-specific survival (CSS) were identified by Cox proportional hazards regression analysis. The IORT and EBRT groups did not differ significantly in OS and CSS. T2 stage (tumor size > 2 cm) was associated with poorer OS (aHR 3.49, 95% CI 1.05-11.62, P = 0.042), whereas ER-positive tumors were associated with better OS (aHR 0.26, 95% CI 0.09-0.76, P = 0.014). IORT was not inferior than EBRT considering the OS and CSS in the short-term follow-up of early breast cancer. It may be a reasonable alternative to EBRT for early breast cancer in select patients with favorable tumor size and receptor status, given the need for long-term monitoring of local control and radiation toxicity.


Experimental observation of the origin and structure of elastoinertial turbulence.
Turbulence generally arises in shear flows if velocities and hence, inertial forces are sufficiently large. In striking contrast, viscoelastic fluids can exhibit disordered motion even at vanishing inertia. Intermediate between these cases, a state of chaotic motion, "elastoinertial turbulence" (EIT), has been observed in a narrow Reynolds number interval. We here determine the origin of EIT in experiments and show that characteristic EIT structures can be detected across an unexpectedly wide range of parameters. Close to onset, a pattern of chevron-shaped streaks emerges in qualitative agreement with linear and weakly nonlinear theory. However, in experiments, the dynamics remain weakly chaotic, and the instability can be traced to far lower Reynolds numbers than permitted by theory. For increasing inertia, the flow undergoes a transformation to a wall mode composed of inclined near-wall streaks and shear layers. This mode persists to what is known as the "maximum drag reduction limit," and overall EIT is found to dominate viscoelastic flows across more than three orders of magnitude in Reynolds number.


[Surgical hand disinfection with "Fesia cito" (author's transl)].
Fürbringer's classical method of scrubbing has been compared with the procedure of embrocation of Fesia cito after Kanz. Concerning the reduction of bacilli both methods are equivalent.


Unveiling the Metal Incorporation Effect of Steady-Active FeP Hydrogen Evolution Nanocatalysts for Water Electrolyzer.
Metal phosphides are promising noble metal-free electrocatalysts for hydrogen evolution reaction (HER), but they usually suffer from inferior stability and thus are far from the device applications. We reported a facile and controllable synthetic method to prepare metal-incorporated M-FeP nanoparticles (M=Cr, Mn, Co, Fe, Ni, Cu, and Mo) with the guide of the density functional theory (DFT). The evaluated HER activity sequence was consistent with the DFT predictions, and cobalt was revealed to be the appropriate dopant. With the optimization of the Co/Fe ratio, the Fe0.67 Co0.33 P/C only required overpotentials of 67 mV and 129 mV to obtain the cathodic current density of 10 and 100 mA cm-2, respectively. It maintained the initial activity in the 10 h stability test, surpassing the other Co-FeP/C catalysts. Ex situ experiments demonstrated that the decreased element leaching and the increased surface phosphide content contributed to the high stability of the Fe0.67 Co0.33 P/C. A proton exchange membrane water electrolyzer was assembled using the Fe0.67 Co0.33 P/C as the cathodic catalyst. It showed a current density of 0.8 A cm-2 at the applied voltage of 2.0 V and retained the initial activity in the 1000 cycles' stability test, suggesting the potential application of the catalysts.


Kurarinone induced p53-independent G0/G1 cell cycle arrest by degradation of K-RAS via WDR76 in human colorectal cancer cells.
Kurarinone (KR), a naturally occurring flavonoid in Sophora flavescens Aiton and a traditional herbal medicine, reportedly has anti-cancer activity against various cancer types both in vitro and in vivo. However, the cellular mechanism of KR remains unknown. Therefore, we aimed to elucidate the mechanism of cell cycle arrest induced by KR in human colorectal cancer cells. KR not only reduced cell proliferation but also induced G0/G1 arrest of colorectal cancer cell lines. The results of western blotting analysis showed that KR reduced the protein levels of cyclin D1/D3 and CDK4/6 by downregulating signaling proteins such as K-RAS, c-MYC, and p-extracellular signal-regulated kinase. Additionally, KR arrested the cell cycle in the G0/G1 phase in a p53-independent manner, and decreased the protein level of K-RAS by proteasomal degradation dependent on WDR76, an E3 ubiquitin ligase. From these results, we propose that KR could be a potent anti-cancer agent, acting through the degradation of K-RAS dependent on WDR76, regardless of the p53 status.


Similar Cost Savings of Bundled Payment Initiatives Applied to Lower Extremity Total Joint Arthroplasty Can Be Achieved Applying Both Models 2 and 3.
In an effort to control cost and increase value, Medicare is transitioning from fee-for-service to value-based alternative payment models (APMs). The Bundled Payments for Care Improvement (BPCI) initiative represents one such voluntary APM. BPCI offers four different bundling options: model 1 covers all Diagnosis Related Groups (DRGs) and Models 2-4 cover 48 clinical episodes, including 186 separate DRGs. The purpose of this investigation is to analyze and compare the cost savings achieved by two different BPCI program participants, provider A and provider B, enrolled in different models of BPCI (Models 2 and 3) for lower extremity joint replacements (LEJRs). We analyzed the BPCI cost savings for Medicare Severity-Diagnosis Related Groups (MS-DRGs) 469 and 470 (lower extremity joint replacement) of two different BPCI program participants. One (provider A) participated in Model 2 while the other (provider B) participated in Model 3. Retrospective payments were based upon savings generated by decreased actual expenses reconciled against target pricing for the episode of care in Models 2 and 3. The Model 2 participant reduced the average cost of all episodes by 18.45%, with all of the savings occurring in the post acute phase. The Model 3 participant reduced episode costs by 16.73%. Both BPCI providers achieved similar cost savings despite participating in different BPCI models. These cost savings all occurred in the post acute setting. The Model 2 provider achieved post acute savings through decreasing overall discharges to institutional post acute care (PAC) providers and decreasing readmissions, while the Model 3 provider decreased costs largely by decreasing the LOS for the institutional PAC providers and decreasing readmissions.


Complex issues with examining diesel exhaust toxicity: is the task getting easier or harder?
Petroleum diesel exhaust (DE) exposure has been linked to several health effects including lung cancer. The role of DE in the cardiopulmonary effects associated with particulate matter (PM) exposures is unclear; this uncertainty drives current research efforts to better understand how the DE exerts toxicity. Several issues present complexities to the design of DE health effects research. One issue is to better establish the health effects and biological responses from DE exposure. Lung responses have been examined with variable findings with controlled exposures, but to date relatively little is known about cardiovascular responses. Additionally, induction of other health effects from DE exposure has been examined mainly through either controlled nonhuman animal model exposures or epidemiological approaches. Due to changing regulations and technology to achieve compliance with regulatory standards, DE from more modern emissions contains less PM and certain gases, making comparisons to older emissions complicated. In addition, the gas phase contains proportionally more mass than the PM phase, presenting technological problems in terms of the collection of the DE for future studies and across-laboratory comparison. Therefore, in order to study the toxicity of DE, whole exhaust (PM and gases) exposures should ideally be performed, at least as part of establishing the role that the PM or gases may play in the biological responses being examined.


Teriflunomide levels in women whose male sexual partner is on teriflunomide for relapsing multiple sclerosis.
For small molecules such as teriflunomide, used to treat relapsing multiple sclerosis (MS), that are potentially embryotoxic, there is a theoretical risk of transmission of the medication from males on the drug to female sexual partners. However, that risk has been undefined up to now. Teriflunomide concentrations were assayed concomitantly in ten sexually active couples, not using barrier methods of contraception, in whom the male partner with MS was on treatment with teriflunomide 14 mg daily for at least two months. These results were compared by male and female age, teriflunomide concentrations and reported average number of incidences of sexual intercourse per month. The threshold level of detection of teriflunomide was 0.020 µg/ml in females. The average age of the cohort was 46.70 for males and 47.10 for females. Four of ten females had detectible teriflunomide concentrations (mean 0.046 µg/ml (range 0.22-0.077, standard deviation 0.025). Male age and both female teriflunomide positive threshold and female teriflunomide concentration were inversely correlated (r = 0.67, R2=0.45, p = 0.034) for the former and (r = 0.62, R2=0.39, p = 0.05, ns) for the latter. No significant correlations were observed for female age, male teriflunomide concentrations, or reported mean monthly episodes of sexual intercourse. This limited study suggests that the small risk that low levels of teriflunomide can be transmitted from male to female partners via sexual intercourse is related to male age. This supports the recommendations found in the United States Product Insert (USPI) stating that men taking teriflunomide who do not wish to father a child, and their female partners, should use reliable contraception. Men wishing to father a child should discontinue use of teriflunomide and undergo an accelerated elimination procedure to reduce the plasma concentrations of the medication to less than 0.02 mg/L (0.02 µg/ml1.


A new series of 2-phenol-4-aryl-6-chlorophenyl pyridine derivatives as dual topoisomerase I/II inhibitors: Synthesis, biological evaluation and 3D-QSAR study.
As a continuous effort to develop novel antitumor agents, a new series of forty-five 2-phenol-4-aryl-6-chlorophenyl pyridine compounds were synthesized and evaluated for cytotoxicity against four different human cancer cell lines (DU145, HCT15, T47D, and HeLa), and topoisomerase I and II inhibitory activity. Several compounds (10-15, 20, 22, 24, 28, 42, and 49) displayed strong to moderate dual topoisomerase I and II inhibitory activity at 100 μM. It was observed that hydroxyl and chlorine moiety at meta or para position of phenyl ring is favorable for dual topoisomerase inhibitory activity and cytotoxicity. Most of the compounds displayed stronger cytotoxicities than those of all positive controls against the HCT15 and T47D cell lines. For investigation of the structure-activity relationships, a 3D-QSAR analysis using the method of comparative molecular field analysis (CoMFA) was performed. The generated 3D contour maps can be used for further rational design of novel terpyridine derivatives as highly selective and potent cytotoxic agents.


Protective agents: regulation of synthesis.
The exogenous cues to overwintering adaptations vary not just between components of hardening but between species. One species, P. brevicornis, initiates glycerol synthesis in response to 0 degree C exposures while a second species, E. solidaginis, increases glycerol levels not in response to temperature but in apparent association with changes in total body mass. This species maintains a constant annual percentage of water while occupying a hibernaculum that dries considerably. During overwintering, E. solidaginis losses approximately 50% of its total body mass. In addition to the changes described, this species (northern populations) increases the amount of water bound to both protein and low-molecular-weight compounds during hardening. The increase in binding exceeds threefold between 25 and -30 degrees C (0.193 to 0.633 g/g dry wt) (29). These data do not unequivocally demonstrate the existence of a hydration trigger to glycerol synthesis but are adequate to put forth such a hypothesis. A decrease in total bulk water levels due to both wet weight loss and increases in bound water may provide conditions of functionally reduced intracellular metabolic water. Since polyol production necessitates the disruption of carbon flow between glucose-6-phosphate and pyruvate, one or more enzymes may be sensitive to water reductions. Pyruvate kinase is sensitive to available water levels. Inhibition of this enzyme would likely cause a shunting of carbon metabolism to glycerol production. This hypothesis becomes attractive in light of the observation that in a variety of species, glycerol accumulations have been correlated with dehydration and hyperosmotic conditions. A common adaptative mechanism may exist in response to apparently different environmental perturbations.


Interrelations between plasma free and protein-bound homocysteine and cysteine in homocystinuria.
To study the interrelations between plasma free and protein-bound homocysteine and cysteine, we measured the levels of these four variables in 167 samples from 17 patients with homocystinuria during different treatment regimens, 14 heterozygotes for cystathionine B-synthase deficiency, and 17 normal subjects. There was a strong positive correlation between free and protein-bound homocysteine, and between free and protein-bound cysteine over a wide range of values in varying clinical situations, but homocysteine and cysteine had differing binding characteristics. At low concentrations homocysteine bound more tightly than cysteine to plasma protein, while at high concentrations of the free amino acids more cysteine than homocysteine was bound to plasma protein. In patients with homocystinuria due to cystathionine B-synthase deficiency, levels of protein-bound homocysteine after an overnight fast were eight to 12 times higher than mean levels +/- SD in the normal subjects of 0.15 +/- 0.03 mumol/g of plasma protein, n = 17, and levels of protein-bound cysteine were lower than mean normal levels +/- SD of 2.30 +/- 0.23 mumol/g, n = 17. In the patients before treatment the proportions of both plasma thiols that were protein-bound were approximately half those in the normal subjects. For homocysteine the proportion was 35% in a representative patient and 78% in normal subjects and in heterozygotes, while for cysteine the corresponding proportions were 26% and 59%. In all blood samples the sum of the plasma free and protein-bound homocysteine and cysteine remained relatively constant (mean +/- SD = 270.6 +/- 68.6 mumol/L of plasma, n = 142).(ABSTRACT TRUNCATED AT 250 WORDS)


Development and single-particle analysis of hybrid extracellular vesicles fused with liposomes using viral fusogenic proteins.
Extracellular vesicles (EVs) have potential biomedical applications, particularly as a means of transport for therapeutic agents. There is a need for rapid and efficient EV-liposome membrane fusion that maintains the integrity of hybrid EVs. We recently described Sf9 insect cell-derived EVs on which functional membrane proteins were presented using a baculovirus-expression system. Here, we developed hybrid EVs by membrane fusion of small liposomes and EVs equipped with baculoviral fusogenic proteins. Single-particle analysis of EV-liposome complexes revealed controlled introduction of liposome components into EVs. Our findings and methodology will support further applications of EV engineering in biomedicine.


Multiple arterial-phase MRI with gadoxetic acid improves diagnosis of hepatocellular carcinoma ≤3.0 cm.
Multiple arterial-phase magnetic resonance imaging (MA-MRI) was introduced to overcome the limitations of gadoxetic acid-enhanced MRI, but its clinical impacts on hepatocellular carcinoma (HCC) diagnosis have not been well assessed. We investigated whether MA-MRI with gadoxetic acid could improve the diagnosis of HCC ≤3.0 cm in comparison with single arterial-phase MRI (SA-MRI). This retrospective study included 397 patients from two tertiary institutions who underwent gadoxetic acid-enhanced MRI (243 patients with 271 lesions in cohort-1 underwent SA-MRI, and 154 patients with 166 lesions in cohort-2 underwent MA-MRI). The patients had 437 hepatic lesions ≤3.0 cm with pathologic confirmation. The arterial-phase image quality and diagnostic performance of SA-MRI and MA-MRI were analysed and compared. To minimize the effects of selection bias because of potential confounding between the two groups, propensity score-matching was additionally performed. MA-MRI showed a significantly higher percentage of optimal arterial-phase timing (94.2% vs. 74.5%, p < .001) and lower incidence of inadequate examinations (1.3% vs. 5.8%, p = .034) than SA-MRI. MA-MRI had a significantly higher non-rim arterial-phase hyperenhancement (APHE) detection rate (94.9% vs. 85.5%, p = .005) and sensitivity for diagnosing HCC (87.4% vs. 70.0%, p < .001) than SA-MRI, but no significant difference in specificity (92.9% vs. 93.1%, p = .966). In 123 pairs of propensity score-matched patients, MA-MRI had significantly higher sensitivity (89.1% vs. 74.5%, p = .006) than SA-MRI with equal specificity (92.3% vs. 92.3%, p > .999). Compared with SA-MRI, MA-MRI with gadoxetic acid can detect more non-rim APHE and significantly improve sensitivity for diagnosing HCC ≤3.0 cm, without a significant decrease in specificity.


Assessing adherence to a rotary diversified diet, a treatment for 'environmental illness'.
To develop and test a method to assess adherence to a rotary diversified diet (RDD), a treatment for environmental illness, which is a putative disorder characterized by multiple sensitivities to foods, chemicals, or inhalants. The RDD requires the elimination of prohibited foods and rotation of remaining nonprohibited foods and their "food families" within a 4- to 7-day cycle. The regimen has yet to be validated to the satisfaction of the scientific community. Details of the 2 components of the RDD prescription, elimination and rotation, were documented, and a food record method of assessing adherence was developed. Adherence to the RDD was then assessed in a cohort of women who were enrolled in a larger prospective study. Test-retest reliability of the adherence assessment method was determined by calculating ratings twice on the same set of patient food records, with 1 week between trials. All patients were contacted through a private environmental medicine clinic in Toronto, Canada. Eight patients provided the food records needed for development of the method; adherence was then assessed in 22 women aged 25 to 67 years. Means, standard deviations, and 95% confidence intervals for adherence ratings were calculated. The reliability of the adherence assessment method was determined by calculating Pearson correlation coefficients for adherence ratings from each trial. A paired t test was also used to determine if the mean differences in ratings between trials were significant. Patients experienced difficulties following both components of the RDD: 37% to 44% of foods consumed were either prohibited or allowed, but were consumed on the incorrect day. The adherence assessment method was found to have high levels of reliability. The adherence assessment method can be used in future evaluations of the RDD, although further testing of the method is recommended. Increased involvement of dietitians with patients diagnosed with environmental illness is recommended.


Task modulated brain connectivity of the amygdala: a meta-analysis of psychophysiological interactions.
Understanding functional connectivity of the amygdala with other brain regions, especially task modulated connectivity, is a critical step toward understanding the role of the amygdala in emotional processes and the interactions between emotion and cognition. The present study performed coordinate-based meta-analysis on studies of task modulated connectivity of the amygdala which used psychophysiological interaction (PPI) analysis. We first analyzed 49 PPI studies on different types of tasks using activation likelihood estimation (ALE) meta-analysis. Widespread cortical and subcortical regions showed consistent task modulated connectivity with the amygdala, including the medial frontal cortex, bilateral insula, anterior cingulate, fusiform gyrus, parahippocampal gyrus, thalamus, and basal ganglia. These regions were in general overlapped with those showed coactivations with the amygdala, suggesting that these regions and amygdala are not only activated together, but also show different levels of interactions during tasks. Further analyses with subsets of PPI studies revealed task specific functional connectivities with the amygdala that were modulated by fear processing, face processing, and emotion regulation. These results suggest a dynamic modulation of connectivity upon task demands, and provide new insights on the functions of the amygdala in different affective and cognitive processes. The meta-analytic approach on PPI studies may offer a framework toward systematical examinations of task modulated connectivity.