The effect of scopolamine and physostigmine on working and reference memory in pigeons.
A comparison of the effects of scopolamine and physostigmine on working memory and reference memory in White Carneaux pigeons was undertaken. In Experiment 1, the pigeons received injections of scopolamine hydrobromide (0.03 mg/kg), or saline. Scopolamine hydrobromide had greater disruptive effects on working memory trials than on reference memory trials, and the centrally active form of scopolamine disrupted working memory trial accuracy more than the peripherally active form. The differential sensitivity of accuracy on working memory trials to disruption by central cholinergic blockade was obtained even though the discrimination required on reference memory trials was more difficult. In Experiment 2, the pigeons received injections of scopolamine hydrobromide (0.015 mg/kg), physostigmine (0.075 mg/kg) both scopolamine and physostigmine, or saline. Physostigmine given with scopolamine was able to reverse the scopolamine-induced reduction of accuracy on working memory trials. In neither study did scopolamine promote accelerated forgetting as the delay interval was increased. These results indicate that manipulation of central cholinergic neurotransmitter systems influences working memory processes in the pigeon, but these effects occur without alterations in the ability of the birds to actively maintain information during the retention interval.


Lipid order, saturation and surface property relationships: a study of human meibum saturation.
Tear film stability decreases with age however the cause(s) of the instability are speculative. Perhaps the more saturated meibum from infants may contribute to tear film stability. The meibum lipid phase transition temperature and lipid hydrocarbon chain order at physiological temperature (33 °C) decrease with increasing age. It is reasonable that stronger lipid-lipid interactions could stabilize the tear film since these interactions must be broken for tear break up to occur. In this study, meibum from a pool of adult donors was saturated catalytically. The influence of saturation on meibum hydrocarbon chain order was determined by infrared spectroscopy. Meibum is in an anhydrous state in the meibomian glands and on the surface of the eyelid. The influence of saturation on the surface properties of meibum was determined using Langmuir trough technology. Saturation of native human meibum did not change the minimum or maximum values of hydrocarbon chain order so at temperatures far above or below the phase transition of human meibum, saturation does not play a role in ordering or disordering the lipid hydrocarbon chains. Saturation did increase the phase transition temperature in human meibum by over 20 °C, a relatively high amount. Surface pressure-area studies showing the late take off and higher maximum surface pressure of saturated meibum compared to native meibum suggest that the saturated meibum film is quite molecularly ordered (stiff molecular arrangement) and elastic (molecules are able to rearrange during compression and expansion) compared with native meibum films which are more fluid agreeing with the infrared spectroscopic results of this study. In saturated meibum, the formation of compacted ordered islands of lipids above the surfactant layer would be expected to decrease the rate of evaporation compared to fluid and more loosely packed native meibum. Higher surface pressure observed with films of saturated meibum compared to native meibum suggests greater film stability especially under the high shear stress of a blink.


Prenatal endogenous androgenic influences on masculine sexual behavior and genital morphology in male and female rats.
Female rats located near a male during uterine development showed increased frequencies of male-like behavior as adults and virilization of genital morphology. These changes in behavior and morphology were blocked by prenatal treatment with the anti-androgen, Flutamide.


Molecular characterization of an EWSR1-POU5F1 fusion associated with a t(6;22) in an undifferentiated soft tissue sarcoma.
We report a soft tissue sarcoma from the thigh with morphologic features resembling Ewing sarcoma, clear cell sarcoma, and myoepithelial tumor of soft tissue. In addition, the genetic and immunohistochemical findings do not correspond to any established pattern, so the tumor does not clearly fit into any one classification. The karyotype analysis revealed a rare chromosomal rearrangement, t(6;22)(p22;q12), that previously has been reported in bone and epithelial tumors. Molecular studies confirmed the presence of an EWSR1-POU5F1 fusion creating a chimeric gene with the N-terminal transcriptional activation domain of EWSR1 and the C-terminal POU DNA binding domain of POU5F1. This report is novel in that to our knowledge, it is the first complete molecular characterization of an EWSR1-POU5F1 fusion in a soft tissue sarcoma. Evaluation of existing data on the known EWSR1-POU5F1 tumors suggests that the fusion gene functions in a wide variety of cell types and may modify the differentiation state of cells, resulting in susceptibility to tumorigenesis.


Assessment of depressive symptoms and functional outcomes in patients with major depressive disorder treated with duloxetine versus placebo: primary outcomes from two trials conducted under the same protocol.
Return of functional ability is a central goal in the treatment of major depressive disorder. We conducted two trials with the same protocol that was designed to assess functioning after 8 Weeks of treatment with duloxetine. The a priori primary outcome was improvement in the Hamilton Depression Rating Scale (HAMD) item 7 (work/activities). Secondary outcomes included improvement in depressive symptoms assessed by the HAMD Maier subscale, and improvement in functioning assessed by the Sheehan Disability Scale (SDS), and the Social Adaptation Self-evaluation Scale (SASS). Patients were randomly assigned to duloxetine 60 mg/day (Trial I, n = 257; Trial II, n = 261) or placebo (Trial I, n = 127; Trial II, n = 131). Changes from baseline were analyzed using a mixed-effects model repeated measures approach. At Week 8, duloxetine was superior to placebo in improving HAMD work/activities (p < 0.001) in Trial II, but not Trial I (p = 0.051), and Maier scores (p < 0.01) in both trials. At Week 12, duloxetine was superior to placebo on improving SASS scores in both trials, and the SDS in Trial II. Treatment with duloxetine was associated with significant improvement in depressive symptoms compared with placebo, but improvement in HAMD work/activities was inconsistent at 8 weeks.


Acute Myocardial Infarction Caused by Spontaneous Coronary Artery Dissection of the First Septal Perforator.
Spontaneous coronary artery dissection of the septal arteries is rare and may be overlooked on coronary angiogram. Additionally, dedicated intracoronary imaging may not be feasible due to artery size. Cardiac magnetic resonance imaging has an emerging role in diagnosis, which is critical because management changes significantly if spontaneous coronary artery dissection is diagnosed. (Level of Difficulty: Beginner.).


Validation of the Adapted Clavien-Dindo in Trauma (ACDiT) Classifications in Medical and Surgical Management of Acute Diverticulitis.
To validate the adapted Clavien-Dindo in trauma (ACDiT) tool as a novel outcome measure for patients with acute diverticulitis managed both operatively and nonoperatively. Complications following diverticulitis are difficult to classify because no traditional tools address patients managed both operatively and nonoperatively. The ACDiT grading system-graded from 0 to 5b-is applied in this manner but has not yet been validated for this patient group. We performed a 5-year observational study of patients with acute diverticulitis at a safety-net hospital. Baseline demographics and hospitalization data were collected. ACDiT scores were assigned, and validation was undertaken by comparing scores with hospital-free days, and verifying that higher scores were associated with known risk factors for poor outcomes. Inverse probability weighted propensity scores were assigned for surgical management, and inverse probability weighted regression analysis was used to determine factors associated with ACDiT ≥ grade 2. Of 260 patients, 188 (72%) were managed nonoperatively. Eighty (31%) developed a complication; 73 (91%) were grades 1 to 3b. Higher grades correlated inversely with hospital-free days (rs = -0.67, P < 0.0001) for all patients and for nonoperative (rs = -0.63, P < 0.0001) and operative (rs = -0.62, P < 0.0001) patients. Hinchey 2 to 3 and initial operative management had higher odds of having a complication of ACDiT ≥ grade 2. The ACDiT tool was successfully applied to acute diverticulitis patients managed operatively and nonoperatively, is associated with known risk factors for adverse outcomes. ACDiT may be considered a meaningful outcome measure for comparing strategies for acute diverticulitis.


Computational Study on Effect of KCNQ1 P535T Mutation in a Cardiac Ventricular Tissue.
Heart diseases such as arrhythmia are the main causes of sudden death. Arrhythmias are typically caused by mutations in specific genes, damage in the cardiac tissue, or due to some chemical exposure. Arrhythmias caused due to mutation is called inherited arrhythmia. Induced arrhythmias are caused due to tissue damage or chemical exposure. Mutations in genes that encode ion channels of the cardiac cells usually result in (dysfunction) improper functioning of the channel. Improper functioning of the ion channel may lead to major changes in the action potential (AP) of the cardiac cells. This further leads to distorted electrical activity of the heart. Distorted electrical activity will affect the ECG that results in arrhythmia. KCNQ1 P535T mutation is one such gene mutation that encodes the potassium ion channel (KV7.1) of the cardiac ventricular tissue. Its clinical significance is not known. This study aims to perform a simulation study on P535T mutation in the KCNQ1 gene that encodes the potassium ion channel KV7.1 in the ventricular tissue grid. The effect of P535T mutation on transmural tissue grids for three genotypes (wild type, heterozygous, and homozygous) of cells are studied and the generated pseudo-ECGs are compared. Results show the delayed repolarization in the cells of ventricular tissue grid. Slower propagation of action potential in the transmural tissue grid is observed in the mutated (heterozygous and homozygous) genotypes. Longer QT interval is also observed in the pseudo-ECG of heterozygous and homozygous genotype tissue grids. From the pseudo-ECGs, it is observed that KCNQ1 P535T mutation leads to Long QT Syndrome (LQTS) which may result in life-threatening arrhythmias, such as Torsade de Pointes (TdP), Jervell and Lange-Nielsen syndrome (JLNS), and Romano-Ward syndrome (RWS).


Gender-dependent differences in degree of facial wrinkles.
This study aimed to reveal gender-dependent differences in the degree of facial wrinkles. Subjects comprised 173 Japanese men and women, divided into four groups according to age. Photographs were taken from nine facial regions and used to classify the intensity of wrinkles into five grades. In addition, replicas were taken from five facial sites and used to measure surface roughness. Data were compared between men and women within each age group. In all age groups, men showed increased forehead wrinkles compared with women. In contrast, no gender-dependent differences were found in upper eyelid wrinkles. Other facial wrinkles were greater in men than in women in all except the oldest group (age, 65-75 years), in which wrinkles in women were greater than or equal to those in men. Our results showed that gender-dependent differences exist in the degree of facial wrinkles. In general, men tend to have more severe wrinkles than women. This tendency disappeared or was reversed in some regions of the face and in individuals more than 60 years old.


Discrimination between sediment and soil samples for forensic purposes using elemental data: an investigation of particle size effects.
This paper reports the results of an investigation to quantify variations in elemental concentrations amongst different particle size fractions obtained from a suite of sediment samples collected from the River Avon, UK. Concentrations of 49 elements determined by inductively coupled plasma spectrometry (ICP-AES and ICP-MS) were compared using conventional and multivariate statistical methods in order to assess compositional differences between different size fractions and between different samples. The results showed significant differences between different size fractions and the bulk sediments, but indicated that all of the size fractions considered (<150, 63-150, 20-63 and <20microm) provided an adequate basis on which to identify associations and differences between samples. It was concluded that, while in certain specific circumstances it may be most appropriate to make forensic comparisons based on a very narrowly defined particle size range, for the majority of purposes the <150microm fraction provides the best compromise between sample size requirements and data resolution.


Yield and In Vitro Antioxidant Potential of Essential Oil from Aerva javanica (Burm. f.) Juss. ex Schul. Flower with Special Emphasis on Seasonal Changes.
The essential oil (EO) of the desert cotton (Aerva javanica (Burm. f.) Juss. ex Schul.) was extracted by hydrodistillation, from A. javanica flowers growing in the sandy soils of the United Arab Emirates (UAE) wild desert. The influence of seasonal variation on flowers' EO yield was studied. The flowers' EO yield obtained from spring samples (0.011%) was significantly the highest followed by early summer (0.009%), winter (0.007%), and autumn samples (0.006%), respectively. The flowers' EO antioxidant analysis were tested by DPPH, FRAP and ABTS assays (in vitro). Results proved that A. javanica flowers' EO, isolated during the four seasons, is a good source of natural bioactive antioxidants. Based on the three tested assays, the highest antioxidant activity was recorded in the spring. Testing of the chemical composition of the flowers' EO was conducted for the season with the highest yield and the best antioxidant performance, recorded in spring, by a combination of gas chromatograph (GC) and gas chromatograph-mass spectrometer (GC-MS). This led to the identification of 29 volatile components, in which the flowers' oil was characterized by angustione as a major compound. Photos by scanning electron microscope (SEM) showed prominent availability of star-shaped trichomes in the epidermis of the flowers.


Effect of sintered silicate-substituted hydroxyapatite on remodelling processes at the bone-implant interface.
Phase pure, sintered granules of hydroxyapatite (HA) and silicon-substituted hydroxyapatite (Si-HA) were implanted for 6 and 12 weeks in an ovine model. Samples containing the bone-implant interface were prepared for ultramicrotomy and transmission electron microscopy (TEM) using an anhydrous sample preparation procedure. The results demonstrate that the morphology of apatite deposits and the sequence of events at the interfaces of bone with pure HA and with Si-HA implants, were different. Organised collagen fibrils were first found at the bone/Si-HA interface after 6 weeks, whereas they were found only after 12 weeks around the pure HA implant. Many more nodular aggregates comprised of plate-like apatite crystallites were observed in the vicinity of Si-HA than around the pure HA after 12 weeks in vivo. These findings suggest that the incorporation of silicate ions into HA promotes processes of bone remodelling at the bone/HA interface. TEM observations suggested that the trabecular bone weaves over the Si-HA and that the collagen fibrils form a mechanical interlock with the Si-HA ceramic implants. High-resolution lattice imaging illustrated apatite crystallites contiguous with the Si-HA ceramic and revealed a direct relationship between the bone mineral and the Si-HA ceramic.


Mechanical Properties and Thermal Conductivity of Thermal Insulation Board Containing Recycled Thermosetting Polyurethane and Thermoplastic.
This study used a mechanochemical method to analyze the recycling mechanism of polyurethane foam and optimize the recycling process. The use of mechanochemical methods to regenerate the polyurethane foam powder breaks the C-O bond of the polyurethane foam and greatly enhances the activity of the powder. Based on orthogonal test design, the mesh, proportion, temperature, and time were selected to produce nine recycled boards by heat pressing. Then, the influence of four factors on the thermal conductivity and tensile strength of the recycled board was analyzed. The results show that 120 mesh polyurethane foam powder has strong activity, and the tensile strength can reach 9.913 Mpa when it is formed at 205 °C and 40 min with 50% PP powder. With the help of the low thermal conductivity of the polyurethane foam, the thermal conductivity of the recycled board can reach 0.037 W/m·K at the parameter of 40 mesh, 80%, 185 °C, 30 min. This research provides an effective method for the recycling of polyurethane foam.


Distribution, ontogeny and projections of cholecystokinin-8, vasoactive intestinal polypeptide and gamma-aminobutyrate-containing neuron systems in the rat spinal cord: an immunohistochemical analysis.
The distribution, ontogeny and fiber projections of cholecystokinin-8, vasoactive intestinal polypeptide and gamma-aminobutyrate-containing neuronal systems in the rat spinal cord were investigated by means of immunocytochemistry. Immunoreactive fibers to cholecystokinin-8, vasoactive intestinal polypeptide and glutamate decarboxylase (gamma-aminobutyrate-synthesizing enzyme, used as a marker of gamma-aminobutyrate) were widely distributed in the spinal cord, being particularly concentrated in the superficial dorsal horn, suggesting a close relationship to the pain transmission system. Cholecystokinin-8-containing neurons were mostly distributed in the dorsal laminae and glutamate decarboxylase-containing neurons were distributed in both the dorsal and ventral horns. Vasoactive intestinal polypeptide-containing neurons were detected in the lateral spinal nucleus and the lamina X. Cholecystokinin-8 and vasoactive intestinal polypeptide immunoreactive structures first appeared on gestational day 17-18. Although no substantial change in immunoreactive structures was observed during the fetal period, they increased markedly after birth. On the other hand, glutamate decarboxylase-positive structures appeared at gestational day 16 and those in the grey matter reached a maximum content at birth; both groups were present in adult animals. Transection of the upper cervical cord resulted in accumulations of cholecystokinin-8 and glutamate decarboxylase rostral to the lesion, revealing the presence of supraspinal projections of cholecystokinin-8 and glutamate decarboxylase to the spinal cord. The same experimental procedure demonstrated the existence of vasoactive intestinal polypeptide-mediating neuronal projections to the supraspinal level, as the accumulating fibers occurred in the area caudal to the lesion.


Kinetics and determinants of the changes of CC16, a lung secretory protein in a rat model of toxic lung injury.
To investigate the kinetics and the determinants of the changes of the Clara cell 16 kDa protein (CC16) as a marker of lung injury following exposure to different toxicants. Rats were treated with 4-Ipomeanol (IPO), alpha-naphtylthiourea (ANTU), sodium chromate (Na(2)CrO(4)) or paraquat-(PQ). The changes of CC16, total protein, albumin, and cystatin-C were determined on BALF supernatan, and CC16, cystatin-C and creatinine levels were also determined in serum. Bronchiolar insult due to IPO or ANTU resulted in an initial transient increase of serum CC16, parallel to that of albumin in BALF, and a subsequent reduction in both BALF and serum. A slight reduction of CC16 in BALF was already apparent one hour following treatment with PQ. In the serum, CC16 increased to 400% of basal value. With PQ and Na(2)CrO(4), the elevation of CC16 was mainly determined by the degree of renal impairment.


Exercise training in older patients with heart failure and preserved ejection fraction: a randomized, controlled, single-blind trial.
Heart failure (HF) with preserved left ventricular ejection fraction (HFPEF) is the most common form of HF in the older population. Exercise intolerance is the primary chronic symptom in patients with HFPEF and is a strong determinant of their reduced quality of life (QOL). Exercise training (ET) improves exercise intolerance and QOL in patients with HF with reduced ejection fraction (EF). However, the effect of ET in HFPEF has not been examined in a randomized controlled trial. This 16-week investigation was a randomized, attention-controlled, single-blind study of medically supervised ET (3 days per week) on exercise intolerance and QOL in 53 elderly patients (mean age, 70±6 years; range, 60 to 82 years; women, 46) with isolated HFPEF (EF ≥50% and no significant coronary, valvular, or pulmonary disease). Attention controls received biweekly follow-up telephone calls. Forty-six patients completed the study (24 ET, 22 controls). Attendance at exercise sessions in the ET group was excellent (88%; range, 64% to 100%). There were no trial-related adverse events. The primary outcome of peak exercise oxygen uptake increased significantly in the ET group compared to the control group (13.8±2.5 to 16.1±2.6 mL/kg per minute [change, 2.3±2.2 mL/kg per minute] versus 12.8±2.6 to 12.5±3.4 mL/kg per minute [change, -0.3±2.1 mL/kg per minute]; P=0.0002). There were significant improvements in peak power output, exercise time, 6-minute walk distance, and ventilatory anaerobic threshold (all P<0.002). There was improvement in the physical QOL score (P=0.03) but not in the total score (P=0.11). ET improves peak and submaximal exercise capacity in older patients with HFPEF.


The Saccharomyces cerevisiae MET3 gene: nucleotide sequence and relationship of the 5' non-coding region to that of MET25.
In Saccharomyces cerevisiae, the expression of several genes implicated in methionine biosynthesis is co-regulated by a specific negative control. To elucidate the molecular basis of this regulation, we have cloned two of these genes, MET3 and MET25. The sequence of MET25 has already been determined (Kerjan et al. 1986). Here, we report the nucleotide sequence of the MET3 gene along with its 5' and 3' flanking regions. Plasmids bearing different deletions upstream of the transcribed region of MET3 were constructed. They were introduced into yeast cells and tested for their ability to complement met3 mutations and to respond to regulation by exogenous methionine. The regulatory region was located within a 100 bp region. The sequence of this regulatory region was compared with that of MET25. A short common sequence which occurs 250-280 bp upstream of the translation initiation codon of the gene was found. This sequence is a good candidate for the cis-acting regulatory element.


Computerized tomography of the eye: a study of 62 pathologic cases.
The usefulness of CT of the eye in diagnosis is discussed, and comparison with ultrasonography is made. The authors draw on experience with 62 patients, including, in particular, cases of tumor, eyeball malformtion, and intraocular foreign bodies. CT is not very useful for study of the eyeball itself, but can be indispensable in the diagnosis of malformations and tumors.


Populations of substance P, Met-enkephalin and serotonin immunoreactive neurons in the interpeduncular nucleus of cat: cytoarchitectonics.
The indirect antibody-peroxidase immunostaining method was used to study the distribution of substance P- (SPLI), Met-enkephalin- (MELI) and serotonin-like (5-HTLI) immunoreactivity within the interpeduncular nuclear complex (IPN) of the cat. Populations of immunoreactive cell bodies were found to have a distinct pattern of localization within the IPN. This distribution pattern is described as seen at 4 representative levels rostrocaudally through the IPN. SPLI cell bodies were mainly at rostral levels I, II and III especially in the median region and as a cluster of cells in the intermediate regions of the IPN. MELI cell bodies were seen at all 4 levels, confined mainly to the median and intermediate regions at levels I and II and also in the lateral regions at levels III and IV. There were few 5-HTLI cell bodies at the rostral levels I and II where they were confined to the dorsal zone of the median region, whereas at levels III and IV there were numerous cell bodies located throughout the intermediate region of the IPN. The immunoreactive cell bodies in the IPN may belong to both intrinsic and projection neurons. The distinctive neurochemical cytoarchitectonics of the IPN suggest that this region of the brain which lies along the course of major limbic pathways, may be a center of complex integrative mechanisms of neuronal activity.


Effects of solvent phase recycling on microalgae liquefaction in ethanol: Bio-oil production and nitrogen transformation.
Liquefaction of microalgae in ethanol offers an eco-friendly bio-oil alternative, but solvent recycling is crucial for sustainability due to extra costs. In this work, Chlorella vulgaris was liquefied in supercritical ethanol at 260 °C, and the solvent phase (SP) separated from bio-oil was recovered and reused. Five liquefaction cycles were performed at identical temperature and pressure conditions to investigate the effects on oil production and nitrogen transformation. The findings demonstrated a gradual increase in water content in recycled SP. Ethanol-water co-solvent as the reaction medium promoted the decomposition and re-polymerization of protein in raw material, thus increasing the bio-oil yield (76.84 %) and higher heating value (33.53 MJ/kg) to some extent. Simultaneously, the relative nitrogen content of bio-oil rose from 8.03 % to 8.52 %, predominantly in the form of nitrogen heterocycles. The potential pathway for nitrogen conversion was revealed, which establishes a theoretical basis for the subsequent denitrification of bio-oil.


NMD Takes the Immune Road to NDD.
Proper mRNA quality control prevents immune activation; when it goes awry, mice and flies develop abnormal behavioral phenotypes. In this issue of Neuron, Johnson et al. (2019) report that inhibiting nonsense-mediated mRNA decay (NMD) contributes to the pathogenesis of neurodevelopmental disorder (NDD) phenotypes by triggering aberrant immune activation.


Insulin Dose and Cardiovascular Mortality in the ACCORD Trial.
In the ACCORD trial, intensive treatment of patients with type 2 diabetes and high cardiovascular (CV) risk was associated with higher all-cause and CV mortality. Post hoc analyses have failed to implicate rapid reduction of glucose, hypoglycemia, or specific drugs as the causes of this finding. We hypothesized that exposure to injected insulin was quantitatively associated with increased CV mortality. We examined insulin exposure data from 10,163 participants with a mean follow-up of 5 years. Using Cox proportional hazards models, we explored associations between CV mortality and total, basal, and prandial insulin dose over time, adjusting for both baseline and on-treatment covariates including randomized intervention assignment. More participants allocated to intensive treatment (79%) than standard treatment (62%) were ever prescribed insulin in ACCORD, with a higher mean updated total daily dose (0.41 vs. 0.30 units/kg) (P < 0.001). Before adjustment for covariates, higher insulin dose was associated with increased risk of CV death (hazard ratios [HRs] per 1 unit/kg/day 1.83 [1.45, 2.31], 2.29 [1.62, 3.23], and 3.36 [2.00, 5.66] for total, basal, and prandial insulin, respectively). However, after adjustment for baseline covariates, no significant association of insulin dose with CV death remained. Moreover, further adjustment for severe hypoglycemia, weight change, attained A1C, and randomized treatment assignment did not materially alter this observation. These analyses provide no support for the hypothesis that insulin dose contributed to CV mortality in ACCORD.


Dynamics of ligand binding to GPCR: Residence time of melanocortins and its modulation.
Ligand binding dynamics and the concept of drug-target residence time are essential factors in the development of novel drugs. Conventional ligand binding assays, which usually collect end-point data, do not provide abundant information regarding the ligand binding kinetics. Therefore, novel methods that allow on-line monitoring of ligand binding processes have to be developed and implemented for drug discovery studies. In this study, we provide a short overview of novel possibilities to characterize ligand binding dynamics to different G protein-coupled receptors (GPCRs). Special attention has been paid to the ligand binding to melanocortin 4 receptors and to the development of a fluorescence anisotropy-based assay system using receptors in budded baculovirus particles. It has been shown that ligand binding to melanocortin 4 receptors occurs to tandemly arranged interconnecting ligand binding sites and that the conventional equilibrium usually cannot be achieved in this system. Therefore, the apparent potencies of the same ligand may differ by up to four orders of magnitude, depending on the experimental conditions and the reporter ligand used.


Functional Analysis of the Minimal Twin-Arginine Translocation System Components from Streptococcus thermophilus CGMCC 7.179 in Escherichia coli DE3.
The twin-arginine translocation (Tat) system, which is used for folded protein secretion, is rare in lactic acid bacteria (LAB). Previously, a Tat system composed of TatAS and TatCS subunits (the subscript S denotes a Streptococcus thermophilus origin) was identified in S. thermophilus CGMCC 7.179. In the present study, the tatA S and tatC S genes were cloned and functionally analyzed in Escherichia coli DE3 tat-deficient mutants. The E. coli tatABCDE-deficient mutant complemented with tatC S A S exhibited shortened cellular chains, but its ability to grow in the presence of sodium dodecyl sulfate (SDS) was not restored, suggesting that the S. thermophilus Tat system could partially replace that of E. coli. Surprisingly, the E. coli tatABE-deficient mutant complemented with tatA S and the E. coli tatC-deficient mutant complemented with tatC S displayed relatively normal cellular morphology and enhanced tolerance to SDS. These results suggest that TatAS and TatCS could serve as active protein translocases in E. coli DE3 tat-deficient mutants. Moreover, TatAS acted as a bifunctional subunit to fulfill the roles of both TatA and TatB of E. coli DE3. Thus, this minimal Tat system would be a promising candidate to translocate recombinant proteins in LAB.


Outcomes of long-term iron supplementation in pediatric restless legs syndrome/periodic limb movement disorder (RLS/PLMD).
Restless legs syndrome (RLS) and periodic limb movement disorder (PLMD) are thought to center around a genetically mediated sensitivity to iron insufficiency. Previous studies have shown the effectiveness of short-term iron therapy in children with low iron storage. Little is known, however, about long-term iron treatment in children with RLS and PLMD. Therefore, we performed this study to assess the long-term effect of iron therapy in children with RLS and PLMD. A retrospective chart review was performed for children who met the following criteria: A) diagnosed as having either RLS or PLMD, B) started on iron supplementation, C) followed up for >2 years in a sleep clinic. Baseline values for iron, ferritin, and periodic limb movement of sleep index (PLMS index) were defined in the three months leading up to the initiation of iron therapy. Values were also computed for follow-up periods of 3-6 months, 1-2 years, and >2 years. Serum iron and ferritin levels and PLMS index were compared between baseline and all subsequent follow-ups. In total, 105 patients met inclusion criteria, of whom 64 were diagnosed with PLMD alone, seven with RLS alone, and 35 with both RLS and PLMD. The average age was 10.2 ± 5.3 years. Compared to the baseline (27.4 ± 12.1 ng/ml), the average ferritin values at 3-6 months (45.62 ± 21.2 ng/ml, p < 0.001, n = 34), 1-2 years (52.0 ± 48.3 ng/ml, p <0.001, n = 63), and >2 years (54.7 ± 40.5 ng/ml, p <0.001, n = 67) were all significantly increased. Inversely, compared to baseline (21 ± 27.0/h, n = 66), PLMS index values at 3-6 months (7.5 ± 9.5/h p < 0.05, n = 11), 1-2 years (6.9 ± 8.9/h, p <0.001, n = 29), and >2 years (10 ± 14.5/h, p <0.001, n = 31) were all significantly decreased. No significant change in serum iron levels was noted at any time point. While retrospective in nature, this study demonstrates a sustained improvement in PLMS index and maintenance of adequate ferritin levels >2 years after iron therapy initiation in our RLS/PLMD cohort with a long-term follow-up. Iron therapy appears to lead to long-lasting improvements in children with RLS/PLMD.


Developing and training mental toughness in sport: a systematic review and meta-analysis of observational studies and pre-test and post-test experiments.
To investigate the efficacy of interventions designed to train and develop mental toughness (MT) in sport. Systematic review and meta-analysis. Journal articles, conference papers and doctoral theses indexed in Embase, Scopus, PubMed and SPORTDiscus from inception to 22 November 2019. Observational and pre-post experimental designs on the efficacy of physical and/or psychological interventions designed to promote MT in athletes. A total of 12 studies, published between 2005 and 2019, were included in the review. A majority of the studies included a sample comprised exclusively of male athletes (54.55%), MT interventions were primarily psychological (83.33%) and most studies measured MT via self-report (75%). The Psychological Performance Inventory (25%), the Mental Toughness Questionnaire-48 (16.67%), and the Mental, Emotional and Bodily Toughness Inventory (16.67%) were the most popular inventories used to measure MT. Methodological quality assessments for controlled intervention studies (k=7), single group pre-test-post-test designs (k=4) and single-subject designs (k=1) indicated that the risk of bias was high in most (75%) of the studies. The meta-analysis involving k=10 studies revealed a large effect (d=0.80, 95% CI 0.30 to 1.28), with variability across studies estimated at 0.56. Although the findings of this review suggest there are effective, empirically based interventions designed to train MT in sport, practitioners should be aware of the level of validity of intervention research before adopting any of the MT training programmes reported in the applied sport psychology literature.


Task Shifting for Initiation and Monitoring of Antiretroviral Therapy for HIV-Infected Adults in Uganda: The SHARE Trial.
With countries moving toward the World Health Organization's "Treat All" recommendation, there is a need to initiate more HIV-infected persons into antiretroviral therapy (ART). In resource-limited settings, task shifting is 1 approach that can address clinician shortages. Uganda. We conducted a randomized controlled trial to test if nurse-initiated and monitored ART (NIMART) is noninferior to clinician-initiated and monitored ART in HIV-infected adults in Uganda. Study participants were HIV-infected, ART-naive, and clinically stable adults. The primary outcome was a composite end point of any of the following: all-cause mortality, virological failure, toxicity, and loss to follow-up at 12 months post-ART initiation. Over half of the study cohort (1,760) was women (54.9%). The mean age was 35.1 years (SD 9.51). Five hundred thirty-three (31.6%) participants experienced the composite end point. At 12 months post-ART initiation, nurse-initiated and monitored ART was noninferior to clinician-initiated and monitored ART. The intention-to-treat site-adjusted risk differences for the composite end point were -4.1 [97.5% confidence interval (CI): = -9.8 to 0.2] with complete case analysis and -3.4 (97.5% CI: = -9.1 to 2.5) with multiple imputation analysis. Per-protocol site-adjusted risk differences were -3.6 (97.5% CI: = -10.5 to 0.6) for complete case analysis and -3.1 (-8.8 to 2.8) for multiple imputation analysis. This difference was within hypothesized margins (6%) for noninferiority. Nurses were noninferior to clinicians for initiation and monitoring of ART. Task shifting to trained nurses is a viable means to increase access to ART. Future studies should evaluate NIMART for other groups (e.g., children, adolescents, and unstable patients).


Task-specific dystonia in hairdressers: a questionnaire survey and review of the literature.
Hairdresser dystonia is one of the occupational dystonias and task-specific movement disorders occurring as a result of long-term repetitive cutting with scissors. The task-specific dystonia manifests itself as a loss of voluntary motor control during extensive practice of cutting requiring a high level of technical proficiency. The prevalence rate of hairdresser dystonia is not well-known worldwide. A questionnaire regarding dystonia was prepared for hairdressers. After sending the questionnaires to 800 hairdressers by direct mail, 134 answers were received by mail. Five of the 134 were suspected to have hairdresser-associated focal dystonia. Thus, 3.7% of hairdressers might have task-specific dystonia. This report was limited because of the small number of participants. However, this research is valuable because it was difficult to find a patient with suspected dystonia due to concerns related to job security.


The effect of a gap concept on peripheral nerve recovery in modified epineurial neurorrhaphy: An experimental study in rats.
Several factors such as surgical approach that only consider topographic anatomy; inadequate fascicular alignment, extraepineurial sprouting in the repair zone; contact of axons with the suture area are the disadvantages of epineurial neurorrhaphy. Accordingly, axonal mismatch, neuroma, and unfavorable nerve recovery become inevitable. Neurotropism is the theory clarifying appropriate matching of the nerve fibers independently without needing surgical approach. The studies comparing the primary nerve repair with the nerve defects bridged in different ways demonstrated better outcomes of nerve recovery in the groups with a nerve gap. In this study, we aimed to demonstrate the effects of the gap concept in primary nerve repair bridged by own epineurium. We hypothesized that this technique will provide better results in terms of peripheral nerve recovery and will significantly eliminate the occurrence of a neuroma, which is quite possible in epineurial neurorrhaphy. A total of 35 Wistar female rats weighing 200 ~ 250 g were randomly divided into five groups each with seven rats. Sham controls constituted Group 1, while the rats with epineural neurorrhaphy were included in Group 2. The remaining three groups were the study groups. In Group 3, after the sciatic nerve transection, epineurium of the distal segment was sleeved and preserved. A 2-mm axonal segment was removed from the epineurium free distal ending and no any procedure was applied to the proximal ending of the transected sciatic nerve. Epineuriums of the both sides were approximated and repaired. In Group 4, a 2-mm axonal segment was removed from the proximal ending of the sciatic nerve after preservation of epineurium and no any procedure was applied to the distal part of sciatic nerve. Epineuriums of the both sides were approximated and repaired. In addition, in Group 5, after epineuriums were sleeved in the both distal and proximal stumps, a 1-mm nerve segment was removed from both endings and epineuriums were repaired in the middle bridging a 2-mm axonal gap again. After a 3 months follow-up period Sciatic Functional Index (SFI) was measured by walking track analysis; the area under the evoked compound muscle action potential (CMAP) and latency periods were calculated via electromyographic (EMG) analysis; and histopathological evaluation were performed to compare the parameters of edema, fibrosis, inflammation, vascularization, axonal degeneration, axonal density, myelination, disorganization, and neuroma occurrence. Vascular structures and nerve fibers were counted at ×200 magnification: +1, +2, and +3 indicated the presence of 0-15, 16-30, and >30 structures, respectively. For uncountable parameters (edema, disorganization, myelination, fibrosis, and inflammation): +1 indicated mild, +2 indicated moderate, and +3 indicated severe. The differences between the groups with axonal gap repair and epineural neurorrhaphy were not significant regarding to SFI. The areas under CMAP were as follows: 27.9 ± 5.9 (Δ = 12.1%) in Group 1; 16.5 ± 5.5 (Δ = 6.3%) in Group 2; 14.1 ± 6.2 (Δ = 4.8%) in group 3; 13.8 ± 2.3 (Δ = 9.2%) in Group 4, and 22.5 ± 18.3 (Δ = 2.2%) in Group 5. Group 5 (1 mm gap in the distal +1 mm gap in the proximal segments) had a significantly better result in terms of the area under CMAP with the value of 22.5 ± 18.3 m/Mv (p = .031). Axonal density was 0.9 ± 0.6 (Δ = 2.2%) in Group 2, 2.4 ± 0.3 (Δ = 5.1%) in Group 3, 2.8 ± 0.1 (Δ = 7.7%) in Group 4, and 2.8 ± 0.2 (Δ = 4.8%) in Group 5. Myelination was 1.1 ± 0.5 (Δ = 3.4%) in group 2, 2.2 ± 0.2 (Δ = 6.7%) in group 3, 2.4 ± 0.4 (Δ = 6.0%) in Group 4, and 2.7 ± 0.3 (Δ = 4.6%) in Group 5. Disorganization was 2.3 ± 0.4 (Δ = 4.1%) in Group 2, 1.2 ± 0.2 (Δ = 7.7%) in Group 3, 1.3 ± 0.2 (Δ = 6.5%) in Group 4, and 1 ± 0.3 (Δ = 5.9%) in Group 5. And, neuroma occurrence was found 2.2 ± 0.6 (Δ = 2.8%) in Group 2 and 0.3 ± 0.2 (Δ = 0.1%) in Group 4 while neuroma was not encountered in Group 3 and Group 5. Comparison between the epineurial neurorrhaphy group and the groups with axonal defect revealed the statistically significant results in the factors of axonal density (p = .001), myelination (p = .028), disorganization (p = .016) and neuroma (p = .001). Creating axonal gap bridged by own epineurium showed favorable results comparing with epineurial neurorrhaphy. Resection of a 1 mm axonal segment from the proximal and distal stumps following the epineurial sleeve procedure and performing the epineurium- only repair can facilitate the nerve regeneration. The feasibility of the described technique has been demonstrated in a small rat model and must be further validated in larger animals before clinical testing.


The urologist as an advocate of men's health: 10 suggested steps toward helping patients achieve better overall health.
Men's health is becoming an important issue. In this review we discuss ways in which urologists can interact more positively with male patients. Ten key messages are set out and it is proposed that the urological specialist could inspire his or her patients by undertaking the same lifestyle changes required to maintain health.


Erratum to "Iterative reconstruction improves image quality and reduces radiation dose in trauma protocols; A human cadaver study".
[This corrects the article DOI: 10.1177/20584601211055389.].


Hairy cell leukemia with bone marrow hypoplasia.
This article reviews the authors' experience with hairy cell leukemia to determine the incidence of hypoplastic marrows in this disease and the utiity of embedding marrow biopsy specimens in plastic, which allows the sections to be stained for tartrate-resistant acid phosphatase (TRAP). It was found that the presence of hairy cells in the peripheral blood, marrow aspirate, or both established the diagnosis in eight of 11 patients; in the remaining three patients, the marrow biopsy specimen was necessary for definitive diagnosis. The bone marrow was markedly hypocellular in three patients, including one in whom the marrow morphologic results were essential for diagnosis. Embedding the marrow biopsy specimen in plastic and staining the sections for TRAP was noted to be a useful technique in establishing the diagnosis of hairy cell leukemia. In all cases in which this was performed, the authors found abundant TRAP-positive cells in the marrow biopsy specimen; even in those cases with markedly hypocellular marrows, this technique revealed the presence of hairy cells. In one patient, TRAP staining of the marrow biopsy specimen proved essential for definitive diagnosis. The authors concluded that the occurrence of marrow hypocellularity is not rare in patients with hairy cell leukemia and that TRAP-staining of plastic-embedded marrow biopsy specimens is a helpful and sometimes essential technique in diagnosing hairy cell leukemia.


The multiple roles of phenols in the degradation of aniline contaminants by sulfate radicals: A combined study of DFT calculations and experiments.
Recent research revealed inhibition or enhancement of dissolved organic matter (DOM) to the degradation of trace organic contaminants (TrOC) in natural and engineered water systems. Phenols containing acetyl, carboxyl, formyl, hydroxy, and methoxy groups were selected as the model DOM to quantitatively study their roles in the degradation of simple anilines, sulfonamide antibiotics, phenylurea pesticides by sulfate radicals (SO4•-). Experimental results found that p-methoxyphenol inhibited aniline and sulfamethoxazole degradation by thermally activated peroxydisulfate (TAP), while p-acetylphenol slightly promoted aniline degradation. Quantum chemical calculations were applied to study the microscopic mechanism and kinetics of phenols affecting the degradation of aniline pollutants (AN) in three ways: competitively reacting with SO4•-, repairing aniline cationic radicals (AN•+) and phenylaminyl radicals (AN(-H)•), and generating phenoxy radicals to degrade anilines. Generally, the degradation of sulfonamides and phenylureas prefer to be inhibited by hydroxy- and methoxy-phenols with low oxidation potential (Eox), due to their diffusion-limiting reaction with SO4•- and rapid back-reduction AN•+ with the calculated rate constants of (0.02 - 6.38) × 109 M-1 s-1. Phenols repairing AN(-H)• through H abstraction reaction is speculated to possibly dominate the joint degradation of phenols and anilines by TAP, which has a poor correlation with Eox. This study provides mechanistic insight into the chemical behavior of complex and heterogeneous DOM in complex aqueous environments.


Hairpin RF resonators for MR imaging transceiver arrays with high inter-channel isolation and B1 efficiency at ultrahigh field 7 T.
Electromagnetic decoupling among a close-fitting or high-density transceiver RF array elements is required to maintain the integrity of the magnetic flux density from individual channel for enhanced performance in detection sensitivity and parallel imaging. High-impedance RF coils have demonstrated to be a prominent design method to circumvent these coupling issues. Yet, inherent characteristics of these coils have ramification on the B1 field efficiency and SNR. In this work, we propose a hairpin high impedance RF resonator design for highly decoupled multichannel transceiver arrays at ultrahigh magnetic fields. Due to the high impedance property of the hairpin resonators, the proposed transceiver array can provide high decoupling performance without using any dedicated decoupling circuit among the resonant elements. Because of elimination of lumped inductors in the resonator circuit, higher B1 field efficiency in imaging subjects can be expected. In order to validate the feasibility of the proposed hairpin RF coils, systematical studies on decoupling performance, field distribution, and SNR are performed, and the results are compared with those obtained from existing high-impedance RF coil, e.g., "self-decoupled RF coil". To further investigate its performance, an 8-channel head coil array using the proposed hairpin resonators loaded with a cylindrical phantom is designed, demonstrating a 19 % increase of the B1+ field intensity compared to the self-decoupled coils at 7 T. Furthermore, the characteristics of the hairpin RF coils are evaluated using a more realistic human head voxel model numerically. The proposed hairpin RF coil provides excellent decoupling performance and superior RF magnetic field efficiency compared to the "self-decoupled" high impedance coils. Bench test of a pair of fabricated hairpin coils prove to be in good accordance with numerical results.


Pharmacodynamic monitoring of BAY 43-9006 (Sorafenib) in phase I clinical trials involving solid tumor and AML/MDS patients, using flow cytometry to monitor activation of the ERK pathway in peripheral blood cells.
We previously reported a flow cytometry technique to monitor pharmacodynamic effects of the raf kinase inhibitor BAY 43-9006 based on the ability of phorbol ester (PMA) to phosphorylate extracellular-regulated kinase (ERK) in peripheral blood (Chow et al., Cytometry 2001;46:72-78). In this article, we describe its application to phase I trials of BAY 43-9006 in solid tumor and AML/MDS patients. The previously described whole blood lysis method was used to monitor BAY 43-9006 effects on peripheral T-cells of solid tumor patients. A modified whole blood fixation protocol was developed for the AML/MDS trial, using the c-kit ligand stem cell factor (SCF) to activate ERK as an alternative to PMA, and incorporating immunophenotypic markers to identify leukemic blasts. At all dose levels of BAY 43-9006 used to treat solid tumor patients, ERK could be activated by PMA in peripheral T-cells and we were not able to show inhibition of raf kinase. A similar effect was seen in the lymphocytes of AML/MDS patients during treatment with BAY 43-9006. However, we found strong inhibition when ERK was activated via c-kit using SCF. Furthermore, normal donor CD34+ve stem cells were much more sensitive to BAY 43-9006 when ERK was activated by SCF, compared to PMA. These findings support the further development of flow cytometry applications to monitor signal transduction inhibitors during early phase clinical trials.


Identification of Conservation Priority Areas and a Protection Network for the Siberian Musk Deer (Moschus moschiferus L.) in Northeast China.
Species conservation actions are guided by available information on the biogeography of the protected species. In this study, we integrated the occurrence data of Siberian musk deer (Moschus moschiferus L.) collected from 2019 to 2021 with species distribution models to estimate the species' potential distribution in Northeast China. We then identified conservation priority areas using a core-area zonation algorithm. In addition, we analyzed core patch fragmentation using FRAGSTATS. Lastly, we identified potential connectivity corridors and constructed a potential protection network based on the least-cost path and the circuit theory. The results showed concentrations of M. moschiferus in the northern Greater Khingan Mountains, the southeastern Lesser Khingan Mountains, and the eastern Changbai Mountains, with a potential distribution area of 127,442.14 km2. Conservation priority areas included 41 core patches with an area of 106,306.43 km2. Patch fragmentation mainly occurred in the Changbai Mountains and the Lesser Khingan Mountains. We constructed an ecological network composed of 41 core patches and 69 linkages for M. moschiferus in Northeast China. The results suggest that the Greater Khingan Mountains represent the most suitable area to maintain the stability of M. moschiferus populations in Northeast China. Considering the high habitat quality requirements of M. moschiferus and its endangered status, we propose that the Chinese government accelerates the construction of the Greater Khingan Mountains National Park and the Lesser Khingan Mountains National Park and enlarges the Northeast China Tiger and Leopard National Park to address the fragmentation of protected areas and the habitat of M. moschiferus.


Technologies for large-scale mapping of functional neural circuits active during a user-defined time window.
The mapping of neural circuits activated during behavior down to individual neurons is crucial for decoding how the brain processes information. Technologies allowing activity-dependent labeling of neurons during user-defined restricted time windows are rapidly developing. Precise marking of the time window with light, in addition to chemicals, is now possible. In these technologies, genetically encoded molecules integrate molecular events resulting from neuronal activity with light/drug-dependent events. The outputs are either changes in fluorescence or activation of gene expression. Molecular reporters allow labeling of activated neurons for visualization and cell-type identification. The transcriptional readout also allows further control of activated neuronal populations using optogenetic tools as reporters. Here we review the design of these technologies and discuss their demonstrated applications to reveal previously unknown connections in the mammalian brain. We also consider the strengths and weaknesses of the current approaches and provide a perspective for the future.


[A comparative analysis of patients with coronary stenotic and ectatic lesions].
A total number of 177 patients with coronary lesions were studied. Sixty nine of them presented either a combination of stenotic and ectatic lesions (n = 18) or isolated ectatic lesions (n = 51) and 108 presented only stenotic lesions. Analyzing several clinical and epidemiological parameters we observed a significant difference with respect to the higher prevalence of hypertension among the group affected by ectatic lesions. We verified as well a higher prevalence of myocardial infarction in the group of ectasia associated to the stenotic lesions. Finally, a higher global mortality was observed in the group of ectasia associated to stenotic lesions after medical or surgical treatment.


Whole-exome sequencing of the mummified remains of Cangrande della Scala (1291-1329 CE) indicates the first known case of late-onset Pompe disease.
Mummified remains of relevant historical figures are nowadays an important source of information to retrace data concerning their private life and health, especially when historical archives are not available. Next-generation-sequencing was proved to be a valuable tool to unravel the characteristics of these individuals through their genetic heritage. Using the strictest criteria currently available for the validation of ancient DNA sequences, whole-genome and whole-exome sequencing were generated from the mummy remains of an Italian nobleman died almost 700 years ago, Cangrande della Scala. While its genome sequencing could not yield sufficient coverage for in depth investigation, exome sequencing could overcome the limitations of this approach to achieve significantly high coverage on coding regions, thus allowing to perform the first extensive exome analysis of a mummy genome. Similar to a standard "clinical exome analysis" conducted on modern DNA, an in-depth variant annotation, high-quality filtering and interpretation was performed, leading to the identification of a genotype associated with late-onset Pompe disease (glycogen storage disease type II). This genetic diagnosis was concordant with the limited clinical history available for Cangrande della Scala, who likely represents the earliest known case of this autosomal recessive metabolic disorder.


A single-step method for the purification of anti-FITC antibodies by use of a coumarin immunosorbent.
Monoclonal anti-fluorescein isothiocyanate (FITC) antibody cross-reacts with 7-hydroxy coumarin derivatives conjugated to BSA. This property permitted the affinity purification of monoclonal anti-FITC antibodies from ascitic fluid using an-immunosorbent consisting of a 7-hydroxy coumarin derivative linked to Sepharose 4B. Ascitic fluid was applied to the immunosorbent column and, after washing, the bound antibody was eluted under extremely mild conditions using 3 M MgCl2. Antibody eluted in this manner was greater than 96% pure as assessed by SDS-PAGE. A polyclonal sheep anti-FITC antibody was also purified from serum on the same immunosorbent to greater than 94% purity. This simple and rapid method for the purification of anti-FITC antibodies will find applications in both immunodiagnostic procedures and in studies of hapten-antibody interactions. The affinity constant of the purified monoclonal anti-FITC antibody conjugated to horseradish peroxidase was assessed by ELISA and was found to be 1.5 x 19(9) M-1.


A neurocomputational theory of action regulation predicts motor behavior in neurotypical individuals and patients with Parkinson's disease.
Surviving in an uncertain environment requires not only the ability to select the best action, but also the flexibility to withhold inappropriate actions when the environmental conditions change. Although selecting and withholding actions have been extensively studied in both human and animals, there is still lack of consensus on the mechanism underlying these action regulation functions, and more importantly, how they inter-relate. A critical gap impeding progress is the lack of a computational theory that will integrate the mechanisms of action regulation into a unified framework. The current study aims to advance our understanding by developing a neurodynamical computational theory that models the mechanism of action regulation that involves suppressing responses, and predicts how disruption of this mechanism can lead to motor deficits in Parkinson's disease (PD) patients. We tested the model predictions in neurotypical individuals and PD patients in three behavioral tasks that involve free action selection between two opposed directions, action selection in the presence of conflicting information and abandoning an ongoing action when a stop signal is presented. Our results and theory suggest an integrated mechanism of action regulation that affects both action initiation and inhibition. When this mechanism is disrupted, motor behavior is affected, leading to longer reaction times and higher error rates in action inhibition.


A new record of the invasive seaweed Caulerpa cylindracea Sonder in the South Adriatic Sea.
The green alga Caulerpa cylindracea Sonder is one of the most infamous and threatening invasive species in the Mediterranean Sea. Since 1985, it started rapidly spreading to all Mediterranean regions causing many ecological changes on natural communities. In the present study, we present an example of this proliferation with the first record in the Marine Protected Area of Tremiti Island (MPATI) in the South Adriatic Sea. Fifteen sites along the coast and 5 different depths have been investigated. Our results provide eveidence of a wide invasion of this pest in three islands, San Domino, San Nicola and Capraia. This study fills a particular data gap in the ongoing biomonitoring of invasive seaweeds in the Mediterranean Sea representing a base line of this invasive species for the MPATI.


Declaration of use and appropriate use of reporting guidelines in high-impact rehabilitation journals is limited: a meta-research study.
The main aims of this metaresearch study conducted among high-impact rehabilitation journals were: 1) to evaluate if the use of reporting guidelines (RGs) was declared and 2) to categorize the declared use as appropriate or inappropriate. Cross-sectional analysis of a random sample of 200 studies published in the period 2010-2019 in five generic rehabilitation journals with the highest 5-year impact factor. Randomized controlled trials, systematic reviews, observational studies, and diagnostic studies were included. Prevalence with 95% confidence intervals (CIs) was estimated for the main outcomes. Among the 200 selected studies, 17.5% (95% CI: 12.2-22.8%) declared using RGs. Among these studies, 48.6% (95% CI: 32-65.1%) declared an appropriate use. There was an increasing trend over time for authors to report the use of RGs (OR: 1.31; 95% CI: 1.13-1.53). Systematic reviews (n = 54) reported more frequently the use of RGs than other study designs (35.2%). In high-impact rehabilitation journals, a small minority of article authors declared the use of RGs. In approximately half of these studies, RGs were used inappropriately. There is an urgent need to improve the use of RGs in this field of research.


Incidental dual source computed tomography imaging of ductal aortic coarctation, left subclavian artery stenosis and bicuspid aortic valve in a patient admitted for atypical chest pain.
A case of incidental ductal aortic coarctation with left subclavian artery stenosis at the origin, severely calcified and stenotic bicuspid aortic valve, and normal coronary arteries demonstrated by single breath hold dual source computed tomography angiography in a 46-year-old man admitted for acute chest pain is presented.


The Dean-Kawasaki Equation and the Structure of Density Fluctuations in Systems of Diffusing Particles.
The Dean-Kawasaki equation-a strongly singular SPDE-is a basic equation of fluctuating hydrodynamics; it has been proposed in the physics literature to describe the fluctuations of the density of N independent diffusing particles in the regime of large particle numbers N≫1. The singular nature of the Dean-Kawasaki equation presents a substantial challenge for both its analysis and its rigorous mathematical justification. Besides being non-renormalisable by the theory of regularity structures by Hairer et al., it has recently been shown to not even admit nontrivial martingale solutions. In the present work, we give a rigorous and fully quantitative justification of the Dean-Kawasaki equation by considering the natural regularisation provided by standard numerical discretisations: We show that structure-preserving discretisations of the Dean-Kawasaki equation may approximate the density fluctuations of N non-interacting diffusing particles to arbitrary order in N-1 (in suitable weak metrics). In other words, the Dean-Kawasaki equation may be interpreted as a "recipe" for accurate and efficient numerical simulations of the density fluctuations of independent diffusing particles.


The protonation state of the cross-linked tyrosine during the catalytic cycle of cytochrome c oxidase.
Cytochrome c oxidase is the terminal complex of the respiratory chain in mitochondria and some aerobic bacteria and is responsible for most of the O(2) consumption in biology. The key reaction in the catalysis of O(2) reduction is O-O bond scission that requires four electrons and a proton. In our recent work (Gorbikova, E. A., Belevich, I., Wikstrom, M., and Verkhovsky, M. I. (2008) Proc. Natl. Acad. Sci. U. S. A. 105, 10733-10737), it was shown that the cross-linked Tyr-280 (Paracoccus denitrificans numbering) provides the proton for O-O bond cleavage. The deprotonated Tyr-280 must be reprotonated later on in the catalytic cycle to serve as a proton donor for the next oxygen reduction event. To find the reaction step at which the cross-linked Tyr-280 becomes reprotonated, all further steps of the catalytic cycle after O-O bond cleavage were followed by infrared spectroscopy. We found that complete reprotonation of the tyrosine is linked to the formation of the one-electron reduced state coupled to reduction of the Cu(B) site.


Effect of Momordica saponin- and Cyclocarya paliurus polysaccharide-enriched beverages on oxidative stress and fat accumulation in Caenorhabditis elegans.
As an edible and medicinal herb in Chinese folk medicine, Cyclocarya paliurus (Batal.) Iljinskaja leaves are traditionally widely used in the treatment of metabolic disorders. The vegetable Momordica charantia L. has been consumed worldwide for thousands of years as a traditional drug due to its activities against obesity and diabetes. In view of the therapeutic value of Momordica saponins (MSs) and C. paliurus polysaccharides (CPPs), an independently developed MSs- and CPPs-containing beverage (MC) was evaluated for its efficacy in controlling oxidative stress and obesity in Caenorhabditis elegans. First, we found that MC could promote the nuclear localization of DAF-16 and the translation of SOD-3. Further exploring its antioxidant properties, the oxidative stress by-products reactive oxygen species, malondialdehyde, and nonesterified fatty acids were significantly inhibited in C. elegans. Moreover, damage due to diseases related to oxidative stress (age pigments and neurodegenerative diseases) was alleviated. Furthermore, fat accumulation was significantly reduced in normal and high-fat models. Finally, the lipid-lowering effects of MC might involve reductions in the size and number of lipid droplets without impairing basic physiological functions in C. elegans. These results provide promising data indicating MC as an innovative health beverage for the pharmacological management of oxidative stress and obesity. © 2020 Society of Chemical Industry.


Effects of IgM from rheumatic fever patients on intracellular calcium levels of neonatal rat cardiac myocytes.
Rheumatic fever (RF), a potential sequela of Streptococcus pyogenes pharyngitis, sometimes results in myocarditis and heart failure. Antibodies have been implicated in the pathogenesis of RF and anti-cardiac myosin antibody levels are elevated in RF patients. Since myocarditis is associated with altered cardiomyocyte calcium transients it was of interest to determine the direct effects of RF patient antibodies on calcium transients in cultured myocytes. RF patient polyclonal IgM treatment caused increased calcium retention by neonatal rat heart cells in vitro as determined with isotopically labeled calcium. Therefore, to further characterize this finding, calcium transients were evaluated by real time fluorescence spectroscopy and deconvolution imaging. RF patient polyclonal IgM produced increased calcium retention during the relaxation stage of the contraction cycle leading to a slowing of contraction rate, disorganized calcium transients, and eventual tetany. In contrast, calcium transient studies of cardiomyocytes following treatment with monoclonal anti-myosin antibodies revealed declining intracellular calcium levels, accompanied by disorganized transients and tetany. Treatment with both antibodies led to myocyte dysfunction and these novel findings suggest a role for antibodies in the pathogenesis of the myocarditis associated with rheumatic carditis.


Regions Important for Hemagglutination Activity and Serotypes of Avibacterium paragallinarum HMTp210 Protein.
Avibacterium paragallinarum is an important respiratory pathogen of domestic chickens. Avibacterium paragallinarum has been subtyped into three serogroups and nine serovars according to the Page and revised Kume schemes. The major hemagglutinin antigen of A. paragallinarum is HMTp210, which is a large protein of about 2000 amino acids (aa), including a 70-aa signal peptide at its N-terminal end. However, the regions important for the hemagglutination (HA) activity and serotypes of HMTp210 remain unclear. In this study we constructed a series of A. paragallinarum strains expressing HMTp210 in-frame deletion mutants and determined their HA titers to identify the regions important for the HA activity and serotypes of HMTp210. Two distinct types of HA activities were found in HMTp210. The type 1 HA activity resided in the region spanning the full-length HA (aa 71-2084), whereas the type 2 resided in the region spanning aa 1003-2084. The putative ligand binding of the type 1 HA activity was located at aa 176-360, which had a structure similar to YadA of Yersinia enterocolitica. The putative ligand binding site of the type 2 HA activity was located at aa 1003-1125, which had a structure similar to UspA1 from Moraxella catarrhalis. The type 1 HA activity appeared to be Page serogroup specific, whereas type 2 appeared to be Kume serovar specific. Finally, sequence analyses of the regions spanning aa 1-400 and aa 1100-1600 of HMTp210 could be useful for the molecular serotyping (the Page and revised Kume schemes) of A. paragallinarum isolates.


Surface antigen expression in chronic lymphocytic leukemia: clustering analysis, interrelationships and effects of chromosomal abnormalities.
Chronic lymphocytic leukemia (CLL) is a phenotypically distinguishable form of B-lymphoid leukemias. The regularity of surface membrane antigen expression patterns, their interrelationships as well as the effects of the three frequent chromosomal aberrations, ie 11q deletion, 13q deletion and trisomy 12, were investigated in 35 classic CLL cases by flow cytometry. The two-way cluster analysis of 31 individual antigens revealed three expression patterns: (1) most cells in most cases positive (CD5, CD19, CD20, CD23, CD27, CD40, CD45, CD45RA); (2) most cells in most cases negative (CD10, CD14, CD34, CD122, CD154, mIgG); and (3) a mixed pattern with a variable number of positive cases and a variable percentage of positive cells in individual cases (CD11c, CD21, CD22, CD25, CD38, CD45RO, CD79b, CD80, CD95, CD124, CD126, CD130, FMC7, mIgD, mIgkappa, mIglambda, mIgM). The expressions of several antigens were strongly interdependent, even when antigens belonged to entirely different gene families. Such antigen pairs were: CD11c/CD21; CD19/CD45; CD19/CD79b; CD22/CD45RA; CD23/Igkappa; CD25/mIgM; CD27/CD45; CD45/CD79b; CD45RA/Igkappa. In contrast, the expression of some antigens was mutually exclusive, the best examples being CD45RA/CD45RO, CD38/CD80 and CD45RA/CD80. Deletion of chromosome arm 11q attenuated expression of splicing variant CD45RA, but enhanced CD45RO expression. In contrast, cases of trisomy 12 were associated with enhanced CD45RA and attenuated CD45RO expression. Similarly, trisomy 12 was associated with enhanced CD27 and mIgkappa expression. The variable levels of signaling surface membrane antigens, their interactions and interference by genetic aberrations are likely to affect the clinical progression and drug response of CLL.


Wilderness medicine: strategies for provision of medical support for adventure racing.
In adventure racing, or multisporting, athletes perform multiple disciplines over a course in rugged, often remote, wilderness terrain. Disciplines may include, but are not limited to, hiking, trail running, mountain biking, caving, technical climbing, fixed-line mountaineering, flat- and white-water boating, and orienteering. While sprint races may be as short as 6 hours, expedition-length adventure races last a minimum of 36 hours up to 10 days or more and may cover hundreds of kilometres. Over the past decade, adventure racing has grown in popularity throughout the world with increasing numbers of events and participants each year. The provision of on-site medical care during these events is essential to ensure the health and safety of the athletes and thus the success of the sport. At present, there are no formal guidelines and a relatively small amount of literature to assist in the development of medical support plans for these events. This article provides an introduction to the provision of medical support for adventure races. Since a wide variety of illness and injury occur during these events, the medical support plan should provide for proper personnel, equipment and supplies to provide care for a wide range of illness and injury. Foot-related problems are the most common reasons for athletes to require medical attention during these events. This article also highlights some of the controversies involved in the provision of medical support for these events. Suggested penalties for acceptance of medical care during the event and strategies for removal of an athlete from the event for medical reasons are offered. In addition, some of the challenges involved in the provision of medical support, including communication, logistics and liability are discussed. This information should prove useful for medical directors of future, similar events. Because of their extreme nature, expedition-length adventure races represent a new and unique area of wilderness and event medicine.


Watercraft decontamination practices to reduce the viability of aquatic invasive species implicated in overland transport.
Recreational boating activities enable aquatic invasive species (AIS) dispersal among disconnected lakes, as invertebrates and plants caught on or contained within watercraft and equipment used in invaded waterbodies can survive overland transport. Besides simple preventive measures such as "clean, drain, dry", resource management agencies recommend decontaminating watercraft and equipment using high water pressure, rinsing with hot water, or air-drying to inhibit this mode of secondary spread. There is a lack of studies assessing the efficacy of these methods under realistic conditions and their feasibility for recreational boaters. Hence, we addressed this knowledge gap via experiments on six invertebrate and plant AIS present in Ontario. Washing at high pressures of 900-1200 psi removed the most biological material (90%) from surfaces. Brief (< 10 s) exposure to water at ≥ 60 °C caused nearly 100% mortality among all species tested, except banded mystery snails. Acclimation to temperatures from 15 to 30 °C before hot water exposure had little effect on the minimum temperature required for no survival. Air-drying durations producing complete mortality were ≥ 60 h for zebra mussels and spiny waterfleas, and ≥ 6 days among plants, whereas survival remained high among snails after a week of air-drying. Hot water exposure followed by air-drying was more effective than either method separately against all species tested.


Epigallocatechin-3-gallate as a potential therapeutic drug for TTR-related amyloidosis: "in vivo" evidence from FAP mice models.
Familial amyloidotic polyneuropathy (FAP) is a neurodegenerative disease caused by the extracellular deposition of mutant transthyretin (TTR), with special involvement of the peripheral nervous system (PNS). Currently, hepatic transplantation is considered the most efficient therapy to halt the progression of clinical symptoms in FAP since more than 95% of TTR is produced by the liver. However, less invasive and more reliable therapeutic approaches have been proposed for FAP therapy, namely based on drugs acting as inhibitors of amyloid formation or as amyloid disruptors. We have recently reported that epigallocatechin-3-gallate (EGCG), the most abundant catechin in green tea, is able to inhibit TTR aggregation and fibril formation, "in vitro" and in a cellular system, and is also able to disrupt pre-formed amyloid fibrils "in vitro". In the present study, we assessed the effect of EGCG subchronic administration on TTR amyloidogenesis "in vivo", using well characterized animal models for FAP. Semiquantitative immunohistochemistry (SQ-IHC) and Western blot analysis of mice tissues after treatment demonstrated that EGCG inhibits TTR toxic aggregates deposition in about 50% along the gastrointestinal tract (GI) and peripheral nervous system (PNS). Moreover EGCG treatment considerably lowered levels of several biomarkers associated with non-fibrillar TTR deposition, namely endoplasmic reticulum (ER)-stress, protein oxidation and apoptosis markers. Treatment of old FAP mice with EGCG resulted not only in the decrease of non-fibrillar TTR deposition but also in disaggregation of amyloid deposits. Consistently, matrix metalloproteinase (MMP)-9 and serum amyloid P component (SAP), both markers of amyloid deposition, were also found reduced in treated old FAP mice. The dual effect of EGCG both as TTR aggregation inhibitor and amyloid fibril disruptor together with the high tolerability and low toxicity of EGCG in humans, point towards the potential use of this compound, or optimized derivatives, in the treatment of TTR-related amyloidoses.


CD123 and More: How to Target the Cell Surface of Blastic Plasmacytoid Dendritic Cell Neoplasm.
Blastic plasmacytoid dendritic cell neoplasm (BPDCN) is a rare and aggressive leukemia derived from plasmacytoid dendritic cells (pDCs). It is associated with a remarkably poor prognosis and unmet need for better therapies. Recently, the first-in-class CD123-targeting therapy, tagraxofusp, was approved for treatment of BPDCN. Other CD123-targeting strategies are in development, including bispecific antibodies and combination approaches with tagraxofusp and other novel agents. In other blood cancers, adoptive T-cell therapy using chimeric antigen receptor (CAR)-modified T cells represents a promising new avenue in immunotherapy, showing durable remissions in some relapsed hematologic malignancies. Here, we report on novel and innovative therapies in development to target surface molecules in BPDCN currently in clinical trials or in preclinical stages. We also discuss new cell surface targets that may have implications for future BPDCN treatment.


Synthesis, Molecular Docking Analysis, and Biological Evaluations of Saccharide-Modified Sulfonamides as Carbonic Anhydrase IX Inhibitors.
Based on the strategy of the "tail approach", 15 novel saccharide-modified sulfonamides were designed and synthesised. The novel compounds were evaluated as inhibitors of three human carbonic anhydrase (CA) isoforms, namely cytoplasmic CA II, transmembrane CA IX, and XII. Most of these compounds showed good activity against CAs and high topological polar surface area (TPSA) values, which had a positive effect on the selective inhibition of transmembrane isoforms CA IX and XII. In the in vitro activity studies, compounds 16a, 16b, and 16e reduced the viability of HT-29 and MDA-MB-231 cells with a high expression of CA IX under hypoxia. The inhibitory activity of compound 16e on the human osteosarcoma cell line MG-63 with a high expression of CA IX and XII was better than that of AZM. Moreover, high concentrations of compounds 16a and 16b reversed the acidification of the tumour microenvironment. In addition, compound 16a had a certain inhibitory effect on the migration of MDA-MB-231 cells. All the above results indicate that the saccharide-modified sulfonamide has further research value for the development of CA IX inhibitors.


Health system capacity for post-abortion care in Java, Indonesia: a signal functions analysis.
The quality of obstetric care has been identified as a contributing factor in Indonesia's persistently high level of maternal mortality, and the country's restrictive abortion laws merit special attention to the quality of post-abortion care (PAC). Due to unique health policies and guidelines, in Indonesia, uterine evacuation for PAC is typically administered only by Ob/Gyns practicing in hospitals. Using data from a survey of 657 hospitals and emergency obstetric-registered public health centers in Java, Indonesia's most populous island, we applied a signal functions analysis to measure the health system's capacity to offer PAC. We then used this framework to simulate the potential impact of the following hypothetical reforms on PAC capacity: allowing first-trimester uterine evacuation for PAC to take place at the primary care level, and allowing provision by clinicians other than Ob/Gyns. Finally, we calculated the proportion of PAC patients treated using four different uterine evacuation procedures. Forty-six percent of hospitals in Java have the full set of services needed to provide PAC, and PAC capacity is concentrated at the highest-level referral hospitals: 86% of referral hospitals have the full set of services, staffing, and equipment compared to 53% of maternity hospitals and 34% of local hospitals. No health centers are adequately staffed or authorized to offer basic PAC services under Indonesia's current guidelines. PAC capacity at all levels of the health system increases substantially in hypothetical scenarios under which authorization to perform first-trimester uterine evacuation for PAC is expanded to midwives and general physicians practicing in health centers. In 2018, 88% percent of PAC patients were treated using dilation and curettage (D&C). Offering first-trimester uterine evacuation for PAC in PONEDs and allowing clinicians other than Ob/Gyns to perform this procedure would greatly improve the capacity of Java's health system to serve PAC patients. Increasing the use of vacuum aspiration and misoprostol for PAC-related uterine evacuation would lower the burden of treatment for patients and facilitate the task-shifting efforts needed to expand access to this life-saving service.


Temperature-dependent variations of 5-hydroxyindoles in ventricular cerebrospinal fluid--an in vivo voltammetric study.
The in vivo voltammetric signal of 5-OH-indoles increased or decreased in the ventricular cerebrospinal fluid when body temperature was artificially increased or decreased in rats in acute experiments. The increase of the signal occurring after raising the body temperature was facilitated by premedication with the monoamine oxidase inhibitor pargyline. It was not abolished by pretreatment with the serotonin synthesis inhibitor para-chlorophenylalanine, although the latter caused an 85% decrease of ventricular 5-OH-indole level. No similar effects were demonstrable in the 5-OH-indole signal of the caudate nucleus recorded simultaneously. An increase of the 5-OH-indole peak was found also in animals chronically implanted with voltammetric electrodes when the ambient temperature was increased in their cages. According to the results of in vitro experiments, a direct physical effect of changes in brain temperature on the sensitivity of the voltammetric electrodes cannot account for changes found in the cerebrospinal fluid. These data, therefore, demonstrate that the increase of body temperature is followed by an increase of 5-OH-indole concentration in the ventricular cerebrospinal fluid. The in vivo voltammetric technique appears to be a suitable method for studying the serotonergic mechanisms involved in thermoregulatory processes.


Bone resorption in osteosclerotic mice is reduced in vitro.
Osteopetrosis in mammals results from a congenital reduction in bone resorption. Calvarial organ cultures were used to measure bone resorption in osteosclerotic (oc/oc) mice and their normal littermates. Measurements of cell-mediated resorption indicate that baseline isotope release by mutant calvariae was only 57% of that observed in normal littermates and isotope release by mutant bone in the presence of parathyroid hormone (PTH) was only 60% of that in normal controls. However, the response of oc calvariae to PTH was not different from normal bone when considered with respect to baseline resorption. These data indicate that bone resorption in oc mice is reduced in both its basal level and in response to PTH and suggest that oc mice are unable to establish normal baseline resorption which may in turn compromise their responsiveness to PTH.


QTL Express: mapping quantitative trait loci in simple and complex pedigrees.
QTL Express is the first application for Quantitative Trait Locus (QTL) mapping in outbred populations with a web-based user interface. User input of three files containing a marker map, trait data and marker genotypes allows mapping of single or multiple QTL by the regression approach, with the option to perform permutation or bootstrap tests.


δ13C-variations of leaves in forests as an indication of reassimilated CO2 from the soil.
An attempt has been made to evaluate the contribution of soil respired CO2 to the total assimilation of a forest tree, by heeding the 13C-concentrations of CO2 from the free atmosphere and from mineralization processes within the soil respectively. An expression has been derived, according to which the assimilated fraction of CO2 from the soil at a particular height of a tree is given by the δ13C-value of the corresponding leaves, δ13C of atmospheric CO2, δ13C of soil respired CO2 and the physiological state of the leaves expressed as the ratio of total respiration over gross photosynthesis and internal over external CO2-concentration. In the particular case investigated, a δ13C-difference of 5‰ has been determined from bottom to top of a beech tree which results in a CO2 contribution from the soil of about 22% for the lower forest strata, while the total contribution of soil respired CO2 accounts for about 5% of the overall assimilation.


Right Lobe Living Donor Hepatectomy in the Setting of Agenesis of Gall Bladder - A Case Report.
Agenesis of Gall Bladder (AGB) is a rare congenital anomaly with only around 500 cases reported so far. The condition may be associated with other biliary anomalies and present diagnostic and technical challenges during hemi hepatectomy which can be surmounted with careful planning. Live donor hepatectomy in the setting of AGB has not been reported before. We report a case of AGB in a potential living donor and highlight the technical modifications used to perform a safe right hepatectomy in this donor.


Brain tissue segmentation in MR images based on a hybrid of MRF and social algorithms.
Effective abnormality detection and diagnosis in Magnetic Resonance Images (MRIs) requires a robust segmentation strategy. Since manual segmentation is a time-consuming task which engages valuable human resources, automatic MRI segmentations received an enormous amount of attention. For this goal, various techniques have been applied. However, Markov Random Field (MRF) based algorithms have produced reasonable results in noisy images compared to other methods. MRF seeks a label field which minimizes an energy function. The traditional minimization method, simulated annealing (SA), uses Monte Carlo simulation to access the minimum solution with heavy computation burden. For this reason, MRFs are rarely used in real time processing environments. This paper proposed a novel method based on MRF and a hybrid of social algorithms that contain an ant colony optimization (ACO) and a Gossiping algorithm which can be used for segmenting single and multispectral MRIs in real time environments. Combining ACO with the Gossiping algorithm helps find the better path using neighborhood information. Therefore, this interaction causes the algorithm to converge to an optimum solution faster. Several experiments on phantom and real images were performed. Results indicate that the proposed algorithm outperforms the traditional MRF and hybrid of MRF-ACO in speed and accuracy.


Rat lens gamma-crystallins. Characterization of the six gene products and their spatial and temporal distribution resulting from differential synthesis.
We have isolated, purified and characterized six individual gamma-crystallin polypeptides present in the rat lens. Comparison of their amino acid compositions with the known structure of the six gamma-crystallin genes permits a one-to-one correspondence to be made between each protein synthesized and the encoding gene. This demonstrates that each of the six genes is actually expressed in vivo. Two classes of three gamma-crystallins each, which we have designated classes gamma ABC and gamma DEF, are known to exist, on the basis of internal sequence homology. We have measured the temperature-dependent phase-separation characteristics of solutions of the six purified gamma-crystallins, and find that the three members of the gamma DEF class (gamma 2-2, gamma 3-1 and gamma 4-1) are all cryo-proteins with relatively high phase-separation temperatures, whereas the three gamma ABC crystallins (gamma 1-1, gamma 1-2 and gamma 2-1) do not show phase separation above -7 degrees C. We have measured the spatial distribution in rat lens of each of the alpha-, beta- and gamma-crystallins as a function of age from 1 to 420 days, using size-exclusion and ion-exchange high-pressure liquid chromatography (HPLC). Our findings in the cortical layer permit us to establish the differential synthesis of each of the crystallins during lens development. Particular attention has been devoted to the spatial and temporal distribution of the six individual gamma-crystallins. Up to birth, synthesis of the three components of the gamma DEF class predominates, and in particular that of gamma 2-2. In subsequent development the three components of the gamma ABC class assume a greater proportion of monomeric crystallins synthesized, while beta s-crystallin synthesis predominates in late development. Our analysis of different layers within single lenses provides novel information on spatial gradients of the water-soluble and water-insoluble protein fractions as a function of age. We consider the consequences of these findings for lens transparency and opacity in both rat and mouse lens. We show that the high concentrations of gamma DEF-crystallins appear to be responsible for the opacity known to occur in young rat lenses. We conclude from these observations that close control of the differential synthesis of gamma-crystallins plays an important role in maintaining lens transparency during development.


Predictors of very early stroke recurrence in the POINT trial population.
Recent trials of acute secondary prevention in patients with minor ischemic stroke or transient ischemic attack (TIA) have demonstrated high rates of early recurrence within days of the initial event. Identifying clinical features associated with early recurrence may guide focused management. Using logistic regression applied to the data of the Platelet Oriented Inhibition in New TIA and Minor Ischemic Stroke (POINT) trial, we evaluated what baseline clinical factors predict outcome events occurring within 7 days of randomization. In the POINT trial, 181 subjects (3.7%) had early recurrence, defined as primary outcome events within 7 days of trial entry, whereas only 100 outcome events occurred over the remainder of the 90 day trial. Protective effects of dual antiplatelet therapy with clopidogrel plus aspirin were seen only as a reduction in these early recurrences, without any impact on later events. In univariate analysis, systolic blood pressure, diastolic blood pressure, serum glucose, initial carotid imaging results, study cohort (minor stroke or TIA), and treatment assignment were significantly associated with early recurrence. Multivariate logistic regression analysis identified a number of factors with significant independent associations with early recurrence, including carotid stenosis or occlusion (Odds Ratio [OR] 2.77; 95% confidence interval [CI] 1.78-4.31), cohort (minor stroke versus TIA) (OR 1.86; 95% CI 1.33-2.58), race (OR 1.57; 95% CI 1.10-2.25), baseline statin use (OR 0.68; 95% CI 0.49-0.95), systolic blood pressure (OR 1.10; 95% CI 1.03-1.18), serum glucose (OR 1.03; 95% CI 1.01-1.05), and age (OR 1.02; 95% CI 1.00-1.03). Receiver Operator Characteristic (ROC) analysis showed a 70% accuracy of the resulting logistic model in predicting early recurrence. Early recurrence is high, and is concentrated in the first 7 days, in patients with minor stroke or TIA. A number of baseline clinical factors, including carotid disease, presentation with minor stroke rather than TIA, race, absence of statin usage, systolic blood pressure, and serum glucose, are independently associated with early event recurrence in the POINT trial population.


Spinal deformity in progressive neuromuscular disease. Natural history and management.
Severe spinal deformity in progressive neuromuscular disease (NMD) leads to multiple problems, including poor sitting balance, difficulty with upright seating and positioning, pain, difficulty in attendant care, and potential exacerbation of underlying restrictive respiratory compromise. Severe scoliosis and pelvic obliquity can in some instances completely preclude upright sitting in a wheelchair. This article reviews the prevalence, natural history, and management of scoliosis in neuromuscular diseases at greatest risk for progressive spinal deformity.


Predominant neuronal B-cell loss in L5 DRG of p75 receptor-deficient mice.
The significance of the p75 low-affinity neurotrophin receptor, for the maintenance and survival of DRG cells, was studied in p75-deficient mice. Perikarya of the L5 DRG of 12-week-old p75 receptor-deficient mice and healthy Balb C mice were compared using stereological techniques. Following systematic sampling, the optical fractionator and the planar vertical rotator were used to estimate the number and mean volume of the cell bodies of the two neuronal subpopulations. The loss of B-cells was 57% (P < 0.00001), numbers being 7300 (CV = 0.12) in controls and 3100 in p75 receptor-deficient mice (CV = 0.18). Also, A-cells showed a significant loss of 39% (P < 0.0001), numbers being 2600 (CV = 0.12) in control mice and 1500 (CV = 0.16) in p75 receptor-deficient mice. The volume of A-cells was reduced by 30% (P<0.01), from 24.700 microm3 (CV=0.17) perikarya in p75 knock-out mice to 15.100 microm3 (CV=0.17) in controls. B-cell volume did not change significantly. It is concluded that the p75 receptor plays a major role in the survival of DRG cells. The predominant loss of small B-cells indicates that the effect of neurotrophins is dependent upon the presence of the p75 low-affinity receptor.


Outcomes of surgery and postoperative radiation therapy in managing medullary thyroid carcinoma.
We evaluated the outcomes of surgery with or without postoperative radiation therapy (PORT) in the management of medullary thyroid carcinoma (MTC). From two tertiary cancer centers, 297 consecutive patients with MTC treated with PORT (n = 46) between 1990 and 2016 or surgery alone (n = 251) between 2000 and 2016 were reviewed. Ten-year cumulative incidences of locoregional and distant failure were 30.2% and 24.9% in the surgery cohort, and 16.9% and 55.2% in the PORT cohort. In the surgery alone cohort, T4 disease, extrathyroidal extension, N1 disease, extranodal extension (ENE), and residual disease after surgery were associated with local failure. The PORT cohort had significantly higher proportions of patients with T4 disease, N1 disease, ENE, and residual disease. High-risk clinical features can help identify patients with MTC at high-risk for local failure after surgery alone. Patients with high-risk clinical features had effective locoregional control after PORT.


Efficacy of whitening toothpaste containing blue covarine: A double-blind controlled randomized clinical trial.
This randomized clinical trial evaluated the efficacy and safety of a blue-covarine whitening toothpaste on tooth bleaching. Seventy-five subjects with shade mean C1 or darker were randomized into three groups (n = 25): CT-conventional toothpaste, WT-whitening toothpaste, and CP10-10% carbamide peroxide. Subjects from CT and WT brushed their teeth twice/day for 2 weeks. CP10 used bleaching gel for 4 hours/night for 2 weeks. Tooth shade and CIELab parameters were measured at baseline, after the first application, 2- and 4-week. ΔE*ab and ΔE00 were calculated. Subjects' perception about tooth color appearance, tooth sensitivity (TS), and gingival irritation (GI) were also assessed. At all evaluation periods, it was not observed differences between WT and CT considering tooth shade (P > 0.7) or CIELab parameters (P > 0.3). At 2- and 4-week, ΔE*ab and ΔE00 were higher for CP10 than WT or CT (P = 0.001). WT and CT reported major dissatisfaction with tooth color appearance than CP10 (P = 0.001). At 1- and 2-week, WT experienced GI and TS similar to CT, both lower than CP10 (P < 0.01). There were no significant differences in the whitening efficacy between whitening and conventional toothpastes. Neither of the dentifrices was as effective as at-home bleaching. The silica-based toothpaste containing blue covarine evaluated in this study did not give significant tooth whitening benefits over 2 weeks of use.


Multiple pregnancy failures: an immunological paradigm.
Recurrent spontaneous abortion (RSA), three or more pregnancy losses prior to 20 weeks, occurs in about 1% of all pregnancies, 50% of RSA cases remain unexplained and unresolved. Recently, immune pathways have been implicated in the pathophysiology of RSA. Immune tolerance of the fetal-placental unit and placental angiogenesis are mandatory for a successful pregnancy outcome. Unscheduled dysregulation of the placental vasculature is thought to be the pathophysiologic mechanisms underlying an array of pregnancy complications like infertility, miscarriage, pre-eclampsia, and fetal growth restriction and death. Investigations on mechanisms and management of RSA are mired by substandard design and lack of optimal randomized clinical trials and have resulted in disagreement on guidelines for evaluation and treatments for patients with multiple pregnancy losses of unknown etiology. The present review focuses on evidence-based research discussion with immunologic causes, and immune-regulatory therapies recommended for helping patients with a history of RSA. We highlight data that might support revalidation of low molecular weight heparin as a protective therapy in RSA. Newly launched growth factors, GM-CSF, and potentially novel agents to suppress inflammatory rejection, including regulatory T cells, human chorionic gonadotropin, and M-CSF/IL-10, may work in concert with tender-loving-care therapy and give hope to couples with multiple pregnancy losses.


A rapid cell membrane permeability test using fluorescent dyes and flow cytometry.
A reliable and rapid test to detect cytotoxic chemicals which affect cell membranes is described. Fluorescein diacetate freely penetrates intact cells where it is hydrolyzed to its fluorochrome, fluorescein, which is retained in the cell due to its polarity. On the other hand, ethidium bromide is known to be excluded from the intact cell, staining only nucleic acids of membrane-damaged cells. The combination of both fluorochromes results in counter-staining: intact cells fluoresce green (cytoplasm) and membrane-damaged cells fluoresce red (nucleus and RNA). Rat thymocytes freshly isolated without enzyme treatment were incubated simultaneously with test substance and dye solution fluorescein diacetate and ethidium bromide. A two-parameter analysis was performed on a flow cytometer with an on-line computer. Concentration-dependent effects of various detergents and solvents were quantified by measuring the amount of dye retention, i.e., the decrease or increase in fluorescein--fluorescence (peak shift), and the decrease in dye exclusion (increase in ethidium bromide-staining) relative to the untreated control. The assay can be used for rapid monitoring of chemical insults to cell membranes which precede the decrease of the viability measured by pure dye exclusion techniques.


MBL2 polymorphisms screening in a regional Italian CF Center.
We performed MBL2 genotyping in 47 CF patients-cared of at the regional CF Centre of Trieste-trying to establish a correlation within allelic variants of MBL2 and modification of patients' clinical outcome. FEV1 values were significantly lowered and a significantly earlier age at onset of Pseudomonas aeruginosa colonisation was found in CF patients with at least one MBL2 variant.