Resveratrol reduces gentamicin-induced EMT in the kidney via inhibition of reactive oxygen species and involving TGF-β/Smad pathway.
Gentamicin (GEN) is one of the most valuable aminoglycoside antibiotics utilized against life-threatening bacterial infections. Unfortunately, GEN-induced nephrotoxicity limited its clinical utility. The pathologic process of nephrotoxicity caused by GEN may involve epithelial to mesenchymal transition (EMT). Resveratrol (RES) is a natural compound was revealed to inhibit EMT in kidney. The present work was conducted to explore the potential renoprotective role of RES on GEN-induced EMT. Moreover, the underlying signaling pathway of this inhibition was investigated. Mice were treated with GEN by intraperitoneal (i.p.) route daily for 15 days to identify EMT onset with regard to GEN-induced nephrotoxicity. To assess the ameliorative role of RES against GEN-induced EMT, RES was i.p. administrated in high and low doses before and concurrently with GEN treatment. GEN administration significantly deteriorated kidney functions. In addition, reduced glutathione (GSH) content and catalase (CAT) activity were significantly decreased with a concomitant increase in the content of kidney malondialdehyde (MDA) after GEN treatment. Histological changes and deposition of collagen were extensive in renal corpuscles and tubules. Increased expression of alpha smooth muscle actin (α-SMA), transforming growth factor-β1 (TGF-β1) and phosphorylated (p)-Smad2 were observed after GEN administration, while E-cadherin expression was decreased. On the contrary, pretreatment with both doses of RES reversed the modifications caused by GEN administration. We concluded that EMT contributes to pathogenesis of GEN-induced nephrotoxicity. RES has a protective effect on GEN-induced EMT via suppressing oxidative stress and a possible involvement of TGF-β/Smad signaling pathway.


[Ultrasonography control of seroma development in patients after radical breast surgery].
Axillary lymphadenectomy, irrespective of volume of surgical intervention, leads to formation of a cavity in the axillary region, its sizes affecting the frequency of postoperative seroma and other complications. Sixty-nine patients with breast cancer underwent ultrasonography for the operative wound, with the latter drained (n-25)/no drain (n = 44). Use of ultrasound for the operative wound permits monitoring the process of seroma formation, carrying out an adequate aspiration of the liquid, avoiding wound draining.


The metrics and correlates of physician migration from Africa.
Physician migration from poor to rich countries is considered an important contributor to the growing health workforce crisis in the developing world. This is particularly true for Africa. The perceived magnitude of such migration for each source country might, however, depend on the choice of metrics used in the analysis. This study examined the influence of choice of migration metrics on the rankings of African countries that suffered the most physician migration, and investigated the correlates of physician migration. Ranking and correlational analyses were conducted on African physician migration data adjusted for bilateral net flows, and supplemented with developmental, economic and health system data. The setting was the 53 African birth countries of African-born physicians working in nine wealthier destination countries. Three metrics of physician migration were used: total number of physician émigrés; emigration fraction defined as the proportion of the potential physician pool working in destination countries; and physician migration density defined as the number of physician émigrés per 1000 population of the African source country. Rankings based on any of the migration metrics differed substantially from those based on the other two metrics. Although the emigration fraction and physician migration density metrics gave proportionality to the migration crisis, only the latter was consistently associated with source countries' workforce capacity, health, health spending, economic and development characteristics. As such, higher physician migration density was seen among African countries with relatively higher health workforce capacity (0.401 < or = r < or = 0.694, p < or = 0.011), health status, health spending, and development. The perceived magnitude of physician migration is sensitive to the choice of metrics. Complementing the emigration fraction, the physician migration density is a metric which gives a different but proportionate picture of which African countries stand to lose relatively more of its physicians with unchecked migration. The nature of health policies geared at health-worker migration can be expected to depend on the choice of migration metrics.


CD27-CD27 ligand/CD70 interactions enhance alloantigen-induced proliferation and cytolytic activity in CD8+ T lymphocytes.
To study the role of CD27-CD27 ligand (CD27L)/CD70 interactions in the generation of murine allospecific T cell responses, SF9 cells or cell membranes expressing recombinant human CD70 were added to in vitro MLC containing C57BL/6 (H-2b) responder cells and class I and II MHC disparate H-2b/d stimulator cells. Alloantigen-specific CTL generation, CD8+ T cell proliferation, and levels of N-alpha-benzyloxycarbonyl-L-lysine thiobenzyl esterase activity were enhanced in the presence of human CD27L/CD70 expressed on SF9 cell membranes. Enhancement of CD8+ T cell responses occurred in the absence of any discernible effects on CD4+ T cell proliferation or IL-2 responses. Additional studies demonstrated that CD27L/CD70-expressing membranes enhanced proliferative responses to class I MHC differences but had no effect on proliferative responses to class II MHC disparities. Enhancement of allospecific CTL generation was also observed when CD27L/CD70-expressing membranes were added only during the last 24 to 48 h of 5-day MLC. Thus, the present studies suggest that CD27-CD27L/CD70 interactions can selectively enhance differentiation of Ag-specific CD8+ T cell effector mechanisms under conditions in which Th cell responses are not altered.


Expression of Human NSAID Activated Gene 1 in Mice Leads to Altered Mammary Gland Differentiation and Impaired Lactation.
Transgenic mice expressing human non-steroidal anti-inflammatory drug activated gene 1 (NAG-1) have less adipose tissue, improved insulin sensitivity, lower insulin levels and are resistant to dietary induced obesity. The hNAG-1 expressing mice are more metabolically active with a higher energy expenditure. This study investigates female reproduction in the hNAG-1 transgenic mice and finds the female mice are fertile but have reduced pup survival after birth. Examination of the mammary glands in these mice suggests that hNAG-1 expressing mice have altered mammary epithelial development during pregnancy, including reduced occupancy of the fat pad and increased apoptosis via TUNEL positive cells on lactation day 2. Pups nursing from hNAG-1 expressing dams have reduced milk spots compared to pups nursing from WT dams. When CD-1 pups were cross-fostered with hNAG-1 or WT dams; reduced milk volume was observed in pups nursing from hNAG-1 dams compared to pups nursing from WT dams in a lactation challenge study. Milk was isolated from WT and hNAG-1 dams, and the milk was found to have secreted NAG-1 protein (approximately 25 ng/mL) from hNAG-1 dams. The WT dams had no detectable hNAG-1 in the milk. A decrease in non-esterified free fatty acids in the milk of hNAG-1 dams was observed. Altered milk composition suggests that the pups were receiving inadequate nutrients during perinatal development. To examine this hypothesis serum was isolated from pups and clinical chemistry points were measured. Male and female pups nursing from hNAG-1 dams had reduced serum triglyceride concentrations. Microarray analysis revealed that genes involved in lipid metabolism are differentially expressed in hNAG-1 mammary glands. Furthermore, the expression of Cidea/CIDEA that has been shown to regulate milk lipid secretion in the mammary gland was reduced in hNAG-1 mammary glands. This study suggests that expression of hNAG-1 in mice leads to impaired lactation and reduces pup survival due to altered milk quality and quantity.


Clinical considerations in anaesthesia for hip arthroplasty.
Experiences with the anaesthetic management of 248 patients undergoing total hip replacement are presented. Blood loss does not appear to be influenced by hypertension, the method of venting or the type of anaesthetic, with the exception of neurolept-analgesia. The importance of oxygen therapy in the treatment of the pulmonary embolic syndrome is stressed and the prevention of deep venous thrombosis is discussed. Mortality and morbidity figures are given.


The impact of multisensory integration and perceptual load in virtual reality settings on performance, workload and presence.
Real-world experience is typically multimodal. Evidence indicates that the facilitation in the detection of multisensory stimuli is modulated by the perceptual load, the amount of information involved in the processing of the stimuli. Here, we used a realistic virtual reality environment while concomitantly acquiring Electroencephalography (EEG) and Galvanic Skin Response (GSR) to investigate how multisensory signals impact target detection in two conditions, high and low perceptual load. Different multimodal stimuli (auditory and vibrotactile) were presented, alone or in combination with the visual target. Results showed that only in the high load condition, multisensory stimuli significantly improve performance, compared to visual stimulation alone. Multisensory stimulation also decreases the EEG-based workload. Instead, the perceived workload, according to the "NASA Task Load Index" questionnaire, was reduced only by the trimodal condition (i.e., visual, auditory, tactile). This trimodal stimulation was more effective in enhancing the sense of presence, that is the feeling of being in the virtual environment, compared to the bimodal or unimodal stimulation. Also, we show that in the high load task, the GSR components are higher compared to the low load condition. Finally, the multimodal stimulation (Visual-Audio-Tactile-VAT and Visual-Audio-VA) induced a significant decrease in latency, and a significant increase in the amplitude of the P300 potentials with respect to the unimodal (visual) and visual and tactile bimodal stimulation, suggesting a faster and more effective processing and detection of stimuli if auditory stimulation is included. Overall, these findings provide insights into the relationship between multisensory integration and human behavior and cognition.


[Endothelial action of aldosterone--therapeutic implications from basic and clinical research].
Aldosterone is the main hormone regulating the water and electrolyte balance of the organism. It is produced and secreted by zona glomerulosa of the adrenal gland cortex, cardiomyocytes as well as by cells of the blood vessel wall. The article is the review of contemporary knowledge of the role of aldosterone in physiology and pathophysiology of cardiovascular system. The authors describe the nuclear and membrane mechanisms of endothelial action of aldosterone, especially their role in the development of cardiovascular system pathologies. New therapeutic indications for aldosterone receptor antagonists are also discussed.


A Natural Experiment Comparing the Effectiveness of the "Healthy Eagles" Child Weight Management Intervention in School Versus Community Settings.
Behavioural weight management interventions are recommended for the treatment of obesity in children. However, the evidence for these is limited and often generated under trial conditions with White, middle-class populations. Healthy Eagles is a behavioural weight management intervention designed to treat excess weight in children. It ran in the London Borough of Croydon from 2017 to 2020 and was delivered in both school and community settings, providing a natural experiment to compare outcomes. A total of 1560 participants started the Healthy Eagles programme; 347 were in the community setting and 703 in the school setting. Data were analysed for those who completed 70% of the programme. In the school setting, there was a small but significant reduction in BMI z-score (M = -0.04, 95% CI = -0.08, -0.01) for participants above a healthy weight, especially in those with severe obesity (M = -0.09, 95% CI = -0.15, -0.03); there was no significant change in any subgroup in the community setting. Linear regression analysis showed the school setting was associated with a 0.26 (95% CI = 0.13, 0.49) greater reduction in BMI z-score than the community setting after adjusting for ethnicity, deprivation, age and gender. Across both programmes, the effect was somewhat greater in participants from a Black (African/Caribbean/Other) ethnic background (M = -0.06, 95% CI = -0.09, -0.02) and from the two most deprived quintiles (M = -0.06, 95% CI = -0.11, -0.01). Data were limited, but minimal changes were measured in nutrition and physical activity behaviours regardless of setting. This evaluation provides indirect evidence of a small but significant benefit to running weight management interventions in a school versus community setting.


A whale of a plastic tale: A plea for interdisciplinary studies to tackle micro- and nanoplastic pollution in the marine realm.
Plastic is one of the most ubiquitous sources of both contamination and pollution of the Anthropocene, and accumulates virtually everywhere on the planet. As such, plastic threatens the environment, the economy and human well-being globally. The related potential threats have been identified as a major global conservation issue and a key research priority. As a consequence, plastic pollution has become one of the most prolific fields of research in research areas including chemistry, physics, oceanography, biology, ecology, ecotoxicology, molecular biology, sociology, economy, conservation, management, and even politics. In this context, one may legitimately expect plastic pollution research to be highly interdisciplinary. However, using the emerging topic of microplastic and nanoplastic leachate (i.e., the desorption of molecules that are adsorbed onto the surface of a polymer and/or absorbed into the polymer matrix in the absence of plastic ingestion) in the ocean as a case study, we argue that this is still far from being the case. Instead, we highlight that plastic pollution research rather seems to remain structured in mostly isolated monodisciplinary studies. A plethora of analytical methods are now available to qualify and quantify plastic monomers, polymers and the related additives. We nevertheless show though a survey of the literature that most studies addressing the effects of leachates on marine organisms essentially still lack of a quantitative assessment of the chemical nature and content of both plastic items and their leachates. In the context of the ever-increasing research effort devoted to assess the biological and ecological effects of plastic waste, we subsequently argue that the lack of a true interdisciplinary approach is likely to hamper the development of this research field. We finally introduce a roadmap for future research which has to evolve through the development of a sound and systematic ability to chemically define what we biologically compare.


Hollow nanoneedle array and its utilization for repeated administration of biomolecules to the same cells.
We present a novel hollow nanoneedle array (NNA) device capable of simultaneously delivering diverse cargo into a group of cells in a culture over prolonged periods. The silica needles are fed by a common reservoir whose content can be replenished and modified in real time while maintaining contact with the same cells. The NNA, albeit its submicrometer features, is fabricated in a silicon-on-insulator wafer using conventional, large scale, silicon technology. 3T3-NIH fibroblast cells and HEK293 human embryonic kidney cells are shown to grow and proliferate successfully on the NNAs. Cargo delivery from the reservoir through the needles to a group of HEK293 cells in the culture is demonstrated by repeated administration of fluorescently labeled dextran to the same cells and transfection with DNA coding for red fluorescent protein. The capabilities demonstrated by the NNA device open the door to large scale studies of the effect of selected cells on their environment as encountered, for instance, in the study of cell-fate decisions, the role of cell-autonomous versus nonautonomous mechanisms in developmental biology, and in the study of excitable cell-networks.


Structure-activity relationship study of first selective inhibitor of excitatory amino acid transporter subtype 1: 2-Amino-4-(4-methoxyphenyl)-7-(naphthalen-1-yl)-5-oxo-5,6,7,8-tetrahydro-4H-chromene-3-carbonitrile (UCPH-101).
The excitatory amino acid transporters (EAATs) are expressed throughout the central nervous system, where they are responsible for the reuptake of the excitatory neurotransmitter (S)-glutamate (Glu). (1) Recently, we have reported the discovery of the first subtype selective EAAT1 inhibitor 2-amino-4-(4-methoxyphenyl)-7-(naphthalen-1-yl)-5-oxo-5,6,7,8-tetrahydro-4H-chromene-3-carbonitrile (UCPH-101) (1b) and presented an introductory structure-activity relationship (SAR) study. (2) Here, we present a detailed SAR by the design, synthesis, and pharmacological evaluation of analogues 1g-1t. By comparison of potencies of 1b, 1h, and 1i versus 1j, it is evident that potency is largely influenced by the chemical nature of the R(1) substituent. The study also demonstrates that any chemical change of the functional groups or a change to the parental scaffold results in the complete loss of inhibitory activity of the compounds at EAAT1. Finally, a bioavailability study of UCPH-101 determined the half-life to be 30 min in serum (rats) but also that it was not able to penetrate the blood-brain barrier to any significant degree.


Direct calculation of 4f(3)-4f(3) transition intensities in Nd(3+)-doped YPO(4) system involving explicit effects of 4f(2)5d configuration.
The effects of the 4f(2)5d configuration on the intraconfigurational [Formula: see text] electric dipole transitions of Nd(3+) doped YPO(4) are taken into account by a 'direct' calculation. A simple model is applied to analyze the opposite-parity 4f(2)5d configuration admixing into 4f(3) transitional states. The matrix elements of the odd-rank crystal-field interaction and the interconfigurational electric dipole transition are directly expressed using a standard tensor operator method. A set of selection rules for f-d mixing and f-f electric dipole transitions is built up. The admixture effect is considered including both explicit 4f(2)5d configuration and other opposite-parity states such as the 4f(2)n(')g configuration which is treated by a closure procedure. Using this calculation method in combination with the experimental data from the absorption spectrum, a set of intensity parameters is obtained. The transition intensities originating from the high-lying (2)G(9/2)(2) level to the lower energy levels are then calculated, demonstrating a good agreement with the experimental results. The new calculation method is suitable for the electric dipole transitions within the 4f(N) configurations of trivalent lanthanide ions with more than two f-electrons.


Hot Adatom Diffusion Following Oxygen Dissociation on Pd(100) and Pd(111): A First-Principles Study of the Equilibration Dynamics of Exothermic Surface Reactions.
We augment ab initio molecular dynamics simulations with a quantitative account of phononic dissipation to study the hyperthermal adsorbate dynamics resulting from a noninstantaneous energy dissipation during exothermic surface chemical reactions. Comparing the hot adatom diffusion ensuing O_{2} dissociation over Pd(100) and Pd(111) we find experimentally accessible product end distances to form a rather misleading measure for the lifetime of this hyperthermal state. The lifetime is particularly long at Pd(111) where a random-walk-type diffusion leads only to small net displacements. A detailed phonon analysis rationalizes the slow equilibration through long-lived Rayleigh mode excitations that spatially confine the released energy within a nanoscopic "hot spot" around the impingement region.


Modulation of antiarrhythmic drug effects by beta-adrenergic sympathetic stimulation.
Appreciation has grown for the impact of the autonomic nervous system on the development of clinical cardiac arrhythmias. Antiarrhythmic medications work to significantly prolong cardiac repolarization and slow conduction. The question has arisen whether these pharmacologic actions of antiarrhythmic drugs can be modulated by alterations in the sympathetic nervous system. This article examines the data pertaining to modulation of the class I and class III effects of antiarrhythmic drugs during beta-adrenergic stimulation, the body's natural response to stress. The actions of several antiarrhythmic drugs can be fully reversed during beta-adrenergic sympathetic stimulation. Overall, the data suggest that pure class III drugs are the most susceptible to reversal of their effects on refractoriness, followed by class IA agents, amiodarone (which has partial resistance), and d,l-sotalol (which is highly resistant to reversal). Whereas retrospective analyses of a number of trials suggest that sympathetic-stimulation-induced reversal of the electrophysiologic effects of certain antiarrhythmic drugs can decrease their clinical efficacy, prospective trials examining this issue are needed. At the current time it appears reasonable to administer beta blockers to patients receiving antiarrhythmic agents that do not have intrinsic antiadrenergic effects.


Epistatic natural allelic variation reveals a function of AGAMOUS-LIKE6 in axillary bud formation in Arabidopsis.
In the Arabidopsis multiparent recombinant inbred line mapping population, a limited number of plants were detected that lacked axillary buds in most of the axils of the cauline (stem) leaves, but formed such buds in almost all rosette axils. Genetic analysis showed that polymorphisms in at least three loci together constitute this phenotype, which only occurs in late-flowering plants. Early flowering is epistatic to two of these loci, called REDUCED SHOOT BRANCHING1 (RSB1) and RSB2, which themselves do not affect flowering time. Map-based cloning and confirmation by transformation with genes from the region where RSB1 was identified by fine-mapping showed that a specific allele of AGAMOUS-Like6 from accession C24 conferred reduced branching in the cauline leaves. Site-directed mutagenesis in the Columbia allele revealed the causal amino acid substitution, which behaved as dominant negative, as was concluded from a loss-of-function mutation that showed the same phenotype in the late-flowering genetic background. This causal allele occurs at a frequency of 15% in the resequenced Arabidopsis thaliana accessions and correlated with reduced stem branching only in late-flowering accessions. The data show the importance of natural variation and epistatic interactions in revealing gene function.


Ovulatory function in epilepsy.
Women with epilepsy have lower fertility rates than women without epilepsy. We hypothesized that limbic dysfunction in temporal lobe epilepsy (TLE) alters the release of hypothalamic trophic hormones that secondarily affect release of the pituitary gonadotropins, causing ovulatory failure. We assessed ovulatory function over three consecutive menstrual cycles in 17 women with partial seizures arising from the temporal lobe (TLE), 7 women with primary generalized epilepsy (PGE), and 12 controls. We devised scores to reflect ovulatory function that were based on daily basal body temperature and monthly serum progesterone levels. Seizure frequency, antiepileptic drugs (AEDs), and depressive symptomatology were also evaluated. Anovulation was more frequent in subjects with TLE (35.3%) than in subjects with PGE (0%) or in controls (8.3%). Anovulatory cycles tended to occur more frequently in subjects with TLE who were treated with polytherapy than in those receiving monotherapy, but this result was not statistically significant. Seizure frequency and symptoms of depression did not affect ovulatory function. Although AED polytherapy may increase the likelihood of anovulation, our results suggest a mechanism of infertility related to temporal lobe dysfunction.


[Littré hernia as a cause of bowel obstruction].
The authors describe a case of a 60-year-old male with a history of a mass and pain at the right inguinal and epigastric areas. He also reported symptoms of bowel obstruction. Physical examination revealed a mass at right inguinal area, which was not reducible or pulsatile. Surgical findings included hernial sac contents with loop of ileum with signs of ischemia and a Meckel's diverticulum. Histopathological examination showed herniation of the Meckel diverticulum -Littré hernia. The article discusses the history and the incidence of this rare form of hernia.


Effects of sinus rhythm maintenance on left heart function after electrical cardioversion of atrial fibrillation: implications for tachycardia-induced cardiomyopathy.
The role of tachycardia-induced cardiomyopathy vs tachycardia-related short diastolic filling period and reduced atrial contraction in decline of left ventricular ejection fraction (LVEF) in atrial fibrillation (AF) is uncertain. We aimed to characterize left heart changes over time in patients with AF who undergo electrical cardioversion (ECV). Consecutive AF patients who were to undergo ECV were enrolled. Patients with unstable or acute heart failure, severe valvular diseases, recent open-heart surgery, major disorders, or an unsuccessful ECV were excluded. Transthoracic echocardiography, including 3-dimensional left atrial and ventricular volume acquisitions, was performed 1-2 hours before and after ECV, and 4-6 weeks later. In 73 patients (77% male, 66 ± 11 years), ECV resulted in an immediate increase in LVEF (from 43 [interquartile range (IQR), 33-50%] to 48 [IQR, 40-53%]; P < 0.0001). Four to 6 weeks after ECV, ejection fraction increased further in patients who remained in sinus rhythm (SR) (n = 55) to 55 (IQR, 44-62)%; P < 0.001. In patients with AF relapse, LVEF returned to values comparable to pre-ECV (n = 18) (44 [IQR, 32-51]%; P = 0.03). The atrial emptying fraction did not significantly change immediately after ECV (n = 69; from 20 [IQR, 13-25]% to 20 [IQR, 15-28]%; P = 0.14). Only patients who remained in SR showed an increase in atrial emptying fraction after 4-6 weeks (n = 51; to 37 [IQR, 26-48]%; P < 0.0001 vs post-ECV). Immediate improvement in LVEF after ECV explains approximately 50% of total LVEF increase over time. However, in SR, LVEF, and atrial function continuously increase over 4-6 weeks after ECV. This might be attributable to recovery of tachycardia-induced cardiomyopathy.


Kava in the treatment of generalized anxiety disorder: a double-blind, randomized, placebo-controlled study.
Kava (Piper methysticum) is a plant-based medicine, which has been previously shown to reduce anxiety. To date, however, no placebo-controlled trial assessing kava in the treatment of generalized anxiety disorder (GAD) has been completed. A total of 75 participants with GAD and no comorbid mood disorder were enrolled in a 6-week double-blind trial of an aqueous extract of kava (120/240 mg of kavalactones per day depending on response) versus placebo. γ-Aminobutyric acid (GABA) and noradrenaline transporter polymorphisms were also analyzed as potential pharmacogenetic markers of response. Reduction in anxiety was measured using the Hamilton Anxiety Rating Scale (HAMA) as the primary outcome. Intention-to-treat analysis was performed on 58 participants who met inclusion criteria after an initial 1 week placebo run-in phase. Results revealed a significant reduction in anxiety for the kava group compared with the placebo group with a moderate effect size (P = 0.046, Cohen d = 0.62). Among participants with moderate to severe Diagnostic and Statistical Manual of Mental Disorders-diagnosed GAD, this effect was larger (P = 0.02; d = 0.82). At conclusion of the controlled phase, 26% of the kava group were classified as remitted (HAMA ≤ 7) compared with 6% of the placebo group (P = 0.04). Within the kava group, GABA transporter polymorphisms rs2601126 (P = 0.021) and rs2697153 (P = 0.046) were associated with HAMA reduction. Kava was well tolerated, and aside from more headaches reported in the kava group (P = 0.05), no other significant differences between groups occurred for any other adverse effects, nor for liver function tests. Standardized kava may be a moderately effective short-term option for the treatment of GAD. Furthermore, specific GABA transporter polymorphisms appear to potentially modify anxiolytic response to kava.


The effect of short-term creatine intake on blood lactic acid and muscle fatigue measured by accelerometer-based tremor response to acute resistance exercise.
The purpose of this study was to investigate the effects of short-term creatine intake on muscle fatigue induced by resistance exercise in healthy adolescent men, i.e., lactic acid concentration and wrist and head tremor measured by an accelerometer. Twelve healthy adolescent men who had no experience with creatine intake were included. The subjects were randomly assigned to the creatine group and the placebo group, followed by 5 days of creatine and placebo intake, and 5 times of 5 sets of leg press, leg extension, bench press, and arm curl exercises at 70% repetition maximum (RM). The lactic acid concentration before and after exercising, rate of perceived exertion (RPE), and accelerometer-based wrist tremor and head tremor during exercise were measured. Subsequently, after 7 days to allow for creatine washout, the same exercise treatment and measurement were performed in each group after switching drug and placebo between the groups. The level of lactic acid before and after the acute resistance exercise trial was significantly lower in the creatine group than in the placebo group (P <0.05). The mean RPE during the resistance exercise was significantly lower in the creatine group than in the placebo group (P <0.05). There was no difference between the two groups in the mean wrist tremor during resistance exercise, but the mean head tremor values were significantly lower in the creatine group than in the placebo group in the arm curl, the last event of the exercise trials (P <0.05). Short-term creatine intake reduces the blood fatigue factor increased by resistance exercise, and is thought to suppress fatigue, especially in the latter half of resistance exercise. Therefore, these findings indicate that short-term creatine intake can have an improved effect on anaerobic exercise performance.


Evaluation of total, ceruloplasmin-associated and type II ferroxidase activities in serum and cerebrospinal fluid of multiple sclerosis patients.
Multiple sclerosis (MS) patients have increased brain iron deposition with higher oxidative stress (OxS). These two features can be caused by an inefficient removal of free iron from extracellular compartment. Ferroxidase activity (Feox) exerted by ceruloplasmin (FeoxCp) and by other molecules (FeoxII) appears to have a central role in this process. The aim of this study was to investigate serum and cerebrospinal fluid (CSF) total Feox, FeoxII and FeoxCp activities in MS patients and neurological controls. Serum and CSF Feox activity, FeoxII and FeoxCp activity was measured in 91 relapsing-remitting (RR) MS patients, 79 subjects with other inflammatory neurological disorders (OIND) and 65 with non-inflammatory neurological disorders (NIND), as controls. This study was approved by the Local Committee for Medical Ethics in Research. Serum total Feox activity was lower in MS group than in both NIND and OIND, with only the former control group differing significantly (p<0.001); FeoxII and FeoxCp activities were comparable among the groups. Serum Feox activities were not associated with disease activity as assessed by clinical examination or by Magnetic Resonance Imaging (MRI). Only total Feox activity was detectable in the CSF and was not different in MS compared to either OIND or NIND. In conclusion, a condition of low systemic Feox may increase the susceptibility of MS patients to iron(II) mediated-oxidative damage. This alteration is not reflected in CSF, suggesting that agents endowed with Feox activity might have different impact in iron homeostasis in the central nervous system compared to periphery.


Estimation of rate constants for serotonin uptake and compartmentation in normal human platelets.
Biochemical and pharmacological experiments led to a model describing a thrombin-releasable (vesicular) compartment and two non-thrombin-releasable compartments for 5-hydroxytryptamine (5-HT, serotonin) in normal human platelets (J. L. Costa, K. L. Kirk, D. L. Murphy, and H. Stark. Br. J. Pharmacol. 72: 449-459, 1981). With tritiated 5-HT as substrate, the amounts of total and nonreleasable intraplatelet tritiated material were measured. Moreover, in the last set of experiments, the amounts of extraplatelet tritiated material and extraplatelet tritiated 5-HT were also measured. With these data, identification of the kinetic constants for movement between the various compartments was possible. A recent extension of the similarity transformation approach to nonlinear compartmental models was used to carry out the a priori identifiability analysis. Then numerical identification was performed by implementing and achieving efficient methods from optimal control theory. Under our experimental conditions, the main results are as follows: 1) the vesicular compartment corresponds to the platelet storage 5-HT compartment, 2) instead of pools I and II proposed previously, only a single nonreleasable thrombin-resistant pool could be detected, and 3) the optimized model computed to suit the first set of experiments fits homologous intraplatelet results obtained under similar experimental conditions in the last set of experiments and also fits one of the two supplementary extraplatelet investigations.


Expression and localization of a UT-B urea transporter in the human bladder.
Facilitative UT-B urea transporters have been shown to play an important role in the urinary concentrating mechanism. Recent studies have now suggested a link between UT-B allelic variation and human bladder cancer risk. UT-B1 protein has been previously identified in the bladder of various mammalian species, but not yet in humans. The aim of the present study was to investigate whether any UT-B protein was present in the human bladder. First, RT-PCR results confirmed that UT-B1 was strongly expressed at the RNA level in the human bladder, whereas UT-B2 was only weakly present. Initial Western blot analysis confirmed that a novel UT-B COOH-terminal antibody detected human UT-B proteins. Importantly, this antibody detected a specific 40- to 45-kDa UT-B signal in human bladder protein. Using a peptide-N-glycosidase F enzyme, this bladder UT-B signal was deglycosylated to a core 30-kDa protein, which is smaller than the predicted size for UT-B1 but similar to many proteins reported to be UT-B1. Finally, immunolocalization experiments confirmed that UT-B protein was strongly expressed throughout all urothelium layers except for the apical membrane of the outermost umbrella cells. In conclusion, these data confirm the presence of UT-B protein within the human bladder. Further studies are now required to determine the precise nature, regulation, and physiological role of this UT-B.


High-Q-factor Al2O3 micro-trench cavities integrated with silicon nitride waveguides on silicon.
We report on the design and performance of high-Q integrated optical micro-trench cavities on silicon. The microcavities are co-integrated with silicon nitride bus waveguides and fabricated using wafer-scale silicon-photonics-compatible processing steps. The amorphous aluminum oxide resonator material is deposited via sputtering in a single straightforward post-processing step. We examine the theoretical and experimental optical properties of the aluminum oxide micro-trench cavities for different bend radii, film thicknesses and near-infrared wavelengths and demonstrate experimental Q factors of > 106. We propose that this high-Q micro-trench cavity design can be applied to incorporate a wide variety of novel microcavity materials, including rare-earth-doped films for microlasers, into wafer-scale silicon photonics platforms.


The association between sporadic somatic parental aneuploidy and chromosomally abnormal placentae in habitual abortions.
To determine whether there is a correlation between the proportion of aneuploid cells in peripheral lymphocytes and the karyotype of the abortus in recurrent miscarriage. 40 couples with recurrent miscarriage and their abortuses were cytogenetically analyzed according to the analysis of 60 cells per proband. Women were divided into two groups according to the proportion of chromosomally abnormal cells in the abortus. Chromosomal analysis was performed using G-banding with trypsin-Giemsa in parental peripheral blood and in the abortus. 20 of the 40 abortuses had a chromosomally abberant karyotype. 65% of parents aborting an embryo with with an increased proportion of chromosomally aberrant cells, had more than 10% aneuploid cells in their peripheral lymphocytes. However, only 12.5% of couples aborting an embryo with chromosomally normal cells had increased rates of aneuploidy. An increased proportion of aneuploid cells was found in the lymphocytes of recurrently aborting couples who repeatedly abort chromosomally abnormal fetuses. Mitotic instability in the lymphocytes may indicate a predisposition to instability at meiosis leading to a chromosomal abberations in the embryo and its subsequent abortion.


First pass effect in posterior circulation occlusions: Analysis from the CICAT registry.
The first pass effect (FPE) is an independent predictor of functional independence in patients with large vessel occlusion in anterior circulation ischemic strokes. However, whether it predicts outcome in posterior circulation large vessel occlusion (PC-LVO) is uncertain. We aimed to study the frequency, characteristics, and predictors of FPE and its association with clinical outcomes in PC-LVO. We performed an analysis from the prospective CICAT Registry. All patients with PC-LVO who underwent endovascular therapy between January 2016 and January 2020 were included. A centrally assessed clinical follow-up was performed at 3 months by blinded investigators. FPE was defined as the achievement of modified Thrombolysis In Cerebral Infarction 3 in a single pass of the endovascular thrombectomy device, and multi-pass effect (MPE) if it was achieved in more than one pass. A multivariable analysis was performed to identify whether FPE is an independent predictor of functional independence defined as a modified Rankin Score of 0-2. We analyzed data from 265 patients in who FPE was achieved in 105 (39.6%). Patients with FPE were more likely to achieve functional independence compared to the non-FPE group (52.4% vs 25.1%, p < .001) and the MPE group (52.4% vs 26.7%, p < .001). FPE was independently associated with functional independence (adjusted odds ratio (aOR): 2.10, 95% confidence interval (CI) 1.01-4.37) but MPE was not (aOR: 0.92, 95% CI 0.40-2.13). Independent predictors of FPE were the use of direct aspiration, embolic mechanism of stroke, and the absence of general anesthesia (GA) use. FPE is an independent predictor of functional independence in PC-LVO and was associated with a significantly better outcome than MPE.


Serum Levels of Meteorin-Like (Metrnl) Are Increased in Patients with Newly Diagnosed Type 2 Diabetes Mellitus and Are Associated with Insulin Resistance.
BACKGROUND Meteorin-like (Metrnl) is a novel adipomyokine that may improve glucose tolerance and affect insulin resistance. This study aimed to investigate the association between serum levels of Metrnl with blood glucose status and to its association with insulin resistance. MATERIAL AND METHODS The study included 160 subjects with normal glucose tolerance (NGT) (n=40), impaired fasting glucose (IFG) (n=40), impaired glucose tolerance (IGT) (n=40), and newly diagnosed type 2 diabetes mellitus (T2DM) (n=40). An enzyme-linked immunosorbent assay (ELISA) was used to measure the serum levels of Metrnl. Partial correlation analysis was used to analyze the relationship between serum levels of Metrnl and metabolic parameters. Multiple logistic regression analysis was performed to identify the association between serum levels of Metrnl with the risk of diabetes. RESULTS Serum levels of Metrnl was highest in patients with T2DM and significantly increased in patients with prediabetes compared with individuals with NGT. After adjusting for age, gender, and body mass index (BMI), serum Metrnl level was significantly correlated with lipid profile, glucose profile, and insulin resistance. Multiple logistic regression analysis showed that Metrnl significantly increased the risk of T2DM (OR=1.727; P=0.008) before adjusting for the homeostatic model assessment of insulin resistance (HOMA-IR). When further adjusted for HOMA-IR, Metrnl was no longer associated with an increased OR for T2DM (OR=1.491; P=0.066), while the HOMA-IR significantly increased the risk of T2DM (OR=1.935; P=0.008). CONCLUSIONS Serum levels of Metrnl were significantly increased in patients with T2DM and may increase the risk of T2DM independent of insulin resistance.


Differential pharmacological properties of beta-adrenoceptor blocking drugs.
After a short introduction on the basic physiology of alpha- and beta-adrenoceptors, including their subdivisions into pre- and postsynaptic subtypes, the principle of competitive antagonism at the level of postjunctional beta-receptor subtypes is explained. Moreover, properties like membrane-stabilizing potency, beta 1-receptor selectivity (cardioselectivity), and intrinsic sympathomimetic activity are discussed at a molecular level. The most frequent applications of beta-adrenoceptor blocking agents in cardiovascular disease, neurology, psychiatry, ophthalmology, and endocrine disorders are enumerated and discussed at a pharmacological level, together with side effects, contraindications, and interactions with other drugs.


Extreme ultraviolet reflectance degradation of aluminum and silicon from surface oxidation.
We have performed in situ oxide contamination and XUV reflectance vs angle of incidence studies on fresh aluminum and silicon films evaporated in an ultrahigh vacuum system (base pressure 2 x 10(-10)Torr). Our ellipsometric measurements indicate that a surface monolayer of oxide forms on aluminum (1 h at 2 x10(-8) Torr oxygen) and silicon (1 h at 10-(7) -Torr oxygen). The monolayer formation time is inversely proportional to oxygen pressure. Our reflectance vs angle of incidence measurements at 58.4-nm wavelength indicate that unoxidized aluminum and silicon coatings can be used as multifacet retroreflectors with net retroreflectances in excess of 75% for aluminum and 50% for silicon.


Work schedule and seasonal influences on sleep and fatigue in helicopter and fixed-wing aircraft operations in extreme environments.
Sleep and fatigue were investigated in aviation search and rescue, firefighting, emergency medical services and offshore transfer operations in 210 participants, for 21 days each, across 17 datasets in seven countries. Sleep data were collected using wrist-worn actigraphs and sleep diaries. Sustained attention was assessed using a 5-min Psychomotor Vigilance Task (PVT). Duty information was provided from corporate IT systems. Despite the number of 24 h operations, most work occurred during daytime hours, and most sleep occurred at night. There were seasonal changes in work and sleep patterns, with naps used to augment total sleep time. The proportion of sleep occurring during duty varied from zero to 30%. Differences in PVT response times were trivial to small. Legislation that defines flight, duty time and minimum rest limits assume that sleep is not obtained during duty periods, apart from some napping under Fatigue Risk Management Systems (FRMS). However, especially in cases where the aviation service requires waiting for tasks (e.g. search and rescue, emergency medical response), this assumption may not always hold. FRMS should accommodate different modes of working that safely facilitate sleep during duty time where appropriate.


Reconstructing web evolution and spider diversification in the molecular era.
The evolutionary diversification of spiders is attributed to spectacular innovations in silk. Spiders are unique in synthesizing many different kinds of silk, and using silk for a variety of ecological functions throughout their lives, particularly to make prey-catching webs. Here, we construct a broad higher-level phylogeny of spiders combining molecular data with traditional morphological and behavioral characters. We use this phylogeny to test the hypothesis that the spider orb web evolved only once. We then examine spider diversification in relation to different web architectures and silk use. We find strong support for a single origin of orb webs, implying a major shift in the spinning of capture silk and repeated loss or transformation of orb webs. We show that abandonment of costly cribellate capture silk correlates with the 2 major diversification events in spiders (1). Replacement of cribellate silk by aqueous silk glue may explain the greater diversity of modern orb-weaving spiders (Araneoidea) compared with cribellate orb-weaving spiders (Deinopoidea) (2). Within the "RTA clade," which is the sister group to orb-weaving spiders and contains half of all spider diversity, >90% of species richness is associated with repeated loss of cribellate silk and abandonment of prey capture webs. Accompanying cribellum loss in both groups is a release from substrate-constrained webs, whether by aerially suspended webs, or by abandoning webs altogether. These behavioral shifts in silk and web production by spiders thus likely played a key role in the dramatic evolutionary success and ecological dominance of spiders as predators of insects.


A redox-linked novel pathway for arsenic-mediated RET tyrosine kinase activation.
We examined the biochemical effects of arsenic on the activities of RET proto-oncogene (c-RET protein tyrosine kinases) and RET oncogene (RET-MEN2A and RET-PTC1 protein tyrosine kinases) products. Arsenic activated c-RET kinase with promotion of disulfide bond-mediated dimerization of c-RET protein. Arsenic further activated RET-MEN2A kinase, which was already 3- to 10-fold augmented by genetic mutation compared with c-RET kinase activity, with promotion of disulfide bond-mediated dimerization of RET-MEN2A protein (superactivation). Arsenic also increased extracellular domain-deleted RET-PTC1 kinase activity with promotion of disulfide bond-mediated dimerization of RET-PTC1 protein. Arsenic increased RET-PTC1 kinase activity with cysteine 365 (C365) replaced by alanine with promotion of dimer formation but not with cysteine 376 (C376) replaced by alanine. Our results suggest that arsenic-mediated regulation of RET kinase activity is dependent on conformational change of RET protein through modulation of a special cysteine sited at the intracellular domain in RET protein (relevant cysteine of C376 in RET-PTC1 protein). Moreover, arsenic enhanced the activity of immunoprecipitated RET protein with increase in thiol-dependent dimer formation. As arsenic (14.2 microM) was detected in the cells cultured with arsenic (100 microM), direct association between arsenic and RET in the cells might modulate dimer formation. Thus, we demonstrated a novel redox-linked mechanism of activation of arsenic-mediated RET proto-oncogene and oncogene products.


Genetic modulation of RNA metabolism in Drosophila. II. Coordinate rate change in 4S, 5S, and poly-A associated RNA synthesis.
It has been demonstrated that a particular rDNA-deficient Y chromosome (Y(bbSuVar-5)) increases the rate of ribosomal RNA synthesis in adult testes (Shermoen and Kiefer 1975) and in whole flies (Clark, Strausbaugh and Kiefer 1977). As an initial attempt to explore the molecular basis of this phenomenon, experiments were designed to determine if the rate increase was specific for rRNA as opposed to the other species of RNA. The genotypes used in these studies were car bb/Y(bb-), car bb/Y( bbSuVar-5), and Sam(+) iso. car bb/Y(bbSuVar-5 ) and car bb/Y(bb-) are deficient to the same extent in rDNA and Sam(+) iso is a wild-type stock. Following isotope incorporation, total RNA was extracted by a phenol:chloroform method and separated by polyacrylamide gel electrophoresis. The various RNA species were quantified by UV absorption and their radioactivity determined by gel fractionation and liquid scintillation counting. The resulting data permitted the calculation of a specific activity (i.e., dpm/microg RNA) which was defined as synthetic rate. Polyadenylated RNA was isolated using a poly-U sepharose column and similar rate calculations were made. The data from these studies indicate that the rate of synthesis of all species of RNA examined (28S + 18S, 5S, 4S transfer RNA and polyadenylated RNA) is increased by the presence of the Y(bbSuVar-5) chromosome. Genetic and molecular mechanisms are discussed.


New classification of autoimmune neuropathies based on target antigens and involved domains of myelinated fibres.
Autoimmune neuropathies are named by eponyms, by descriptive terminology or because of the presence of specific antibodies and are traditionally classified, on the basis of pathology and electrophysiology, as primary demyelinating or axonal. However, autoimmune disorders targeting specific molecules of the nodal region, although not showing pathological evidence of demyelination, can exhibit all the electrophysiological changes considered characteristic of a demyelinating neuropathy and acute neuropathies with antiganglioside antibodies, classified as axonal and due to nodal dysfunction, can present with reversible conduction failure and prompt recovery that appear contradictory with the common view of an axonal neuropathy. These observations bring into question the concepts of demyelinating and axonal nerve conduction changes and the groundwork of the classical dichotomous classification.We propose a classification of autoimmune neuropathies based on the involved domains of the myelinated fibre and, when known, on the antigen. This classification, in our opinion, helps to better systematise autoimmune neuropathies because points to the site and molecular target of the autoimmune attack, reconciles some contrasting pathological and electrophysiological findings, circumvents the apparent paradox that neuropathies labelled as axonal may be promptly reversible and finally avoids taxonomic confusion and possible misdiagnosis.


Cannabidiol-Mediated Changes to the Phospholipid Profile of UVB-Irradiated Keratinocytes from Psoriatic Patients.
UVB phototherapy is treatment for psoriasis, which increases phospholipid oxidative modifications in the cell membrane of the skin. Therefore, we carried out lipidomic analysis on the keratinocytes of healthy individuals and patients with psoriasis irradiated with UVB and treated with cannabidiol (CBD), phytocannabinoid with antioxidant and anti-inflammatory properties. Our results showed that, in psoriatic keratinocytes phosphatidylcholine (PC), phosphatidylinositol (PI), phosphatidylserine (PS), and ether-linked phosphoethanolamine (PEo), were downregulated, while SM (d41:2) was upregulated. These changes were accompanied by an increase in negative zeta potential, which indicates translocation of PS to the outer layer of the membrane. CBD treatment of psoriatic keratinocytes led to downregulation of PC, PS, and upregulation of certain PEo and an SM species, SM (d42:2), and the zeta potential. However, UVB irradiation of psoriatic keratinocytes resulted in upregulation of PC, PC plasmalogens (PCp), PEo, and a decrease in the negative zeta potential. The exposure of UVB-irradiated cells to CBD led to a decrease in the level of SM (d42:2). Our results suggest that CBD induces pro-apoptotic mechanisms in psoriatic keratinocytes while simultaneously improving the antioxidant properties and preventing the loss of transepidermal water of keratinocytes of patients irradiated with UVB. Thus, CBD has potential therapeutic value in the treatment of psoriasis.


Localized delivery of immunotherapy via implantable scaffolds for breast cancer treatment.
Breast cancer remains a leading global cause of morbidity and mortality. While the field of immunotherapy is a promising avenue of investigation and has revolutionized the standard of care for melanoma and lung cancer, modest response rates and a high incidence of immune-related adverse events often necessitate the administration of a sub-therapeutic dose or treatment cessation. Injectable and implantable drug delivery devices present a novel strategy to achieve sustained delivery of potent concentrations of drug directly to the tumor site and minimize systemic toxicity. This review will address the current limitations with conventional immunotherapy for breast cancer treatment, and the recent developments and future prospects in localized delivery strategies. We describe implantable scaffolds and injectable biomaterials for the localized delivery of immunotherapy, which can improve the safety and efficacy of immunotherapies. We discuss the limitations of these delivery systems, such as the influence of shape and material type on drug release and tumor uptake. The challenges of clinical translation, such as the availability of appropriate preclinical animal models and accurate reporting are also discussed. Considerations of these issues will pave the way for effective new therapies that will improve treatment response, patient survival and quality of life for breast cancer patients.


Belumosudil for chronic graft-versus-host disease after 2 or more prior lines of therapy: the ROCKstar Study.
Belumosudil, an investigational oral selective inhibitor of Rho-associated coiled-coil-containing protein kinase 2 (ROCK2), reduces type 17 and follicular T helper cells via downregulation of STAT3 and enhances regulatory T cells via upregulation of STAT5. Belumosudil may effectively treat patients with chronic graft-versus-host disease (cGVHD), a major cause of morbidity and late nonrelapse mortality after an allogeneic hematopoietic cell transplant. This phase 2 randomized multicenter registration study evaluated belumosudil 200 mg daily (n = 66) and 200 mg twice daily (n = 66) in subjects with cGVHD who had received 2 to 5 prior lines of therapy. The primary end point was best overall response rate (ORR). Duration of response (DOR), changes in Lee Symptom Scale score, failure-free survival, corticosteroid dose reductions, and overall survival were also evaluated. Overall median follow-up was 14 months. The best ORR for belumosudil 200 mg daily and 200 mg twice daily was 74% (95% confidence interval [CI], 62-84) and 77% (95% CI, 65-87), respectively, with high response rates observed in all subgroups. All affected organs demonstrated complete responses. The median DOR was 54 weeks; 44% of subjects have remained on therapy for ≥1 year. Symptom reduction with belumosudil 200 mg daily and 200 mg twice daily was reported in 59% and 62% of subjects, respectively. Adverse events (AEs) were consistent with those expected in patients with cGVHD receiving corticosteroids and other immunosuppressants. Sixteen subjects (12%) discontinued belumosudil because of possible drug-related AEs. Belumosudil, a promising therapy for cGVHD, was well tolerated with clinically meaningful responses. This trial was registered at www.clinicaltrials.gov as #NCT03640481.


Sorting Nexin 27 Regulates the Lysosomal Degradation of Aquaporin-2 Protein in the Kidney Collecting Duct.
Sorting nexin 27 (SNX27), a PDZ (Postsynaptic density-95/Discs large/Zonula occludens 1) domain-containing protein, cooperates with a retromer complex, which regulates intracellular trafficking and the abundance of membrane proteins. Since the carboxyl terminus of aquaporin-2 (AQP2c) has a class I PDZ-interacting motif (X-T/S-X-Φ), the role of SNX27 in the regulation of AQP2 was studied. Co-immunoprecipitation assay of the rat kidney demonstrated an interaction of SNX27 with AQP2. Glutathione S-transferase (GST) pull-down assays revealed an interaction of the PDZ domain of SNX27 with AQP2c. Immunocytochemistry of HeLa cells co-transfected with FLAG-SNX27 and hemagglutinin (HA)-AQP2 also revealed co-localization throughout the cytoplasm. When the PDZ domain was deleted, punctate HA-AQP2 labeling was localized in the perinuclear region. The labeling was intensively overlaid by Lysotracker staining but not by GM130 labeling, a cis-Golgi marker. In rat kidneys and primary cultured inner medullary collecting duct cells, the subcellular redistribution of SNX27 was similar to AQP2 under 1-deamino-8-D-arginine vasopressin (dDAVP) stimulation/withdrawal. Cell surface biotinylation assay showed that dDAVP-induced AQP2 translocation to the apical plasma membrane was unaffected after SNX27 knockdown in mpkCCD cells. In contrast, the dDAVP-induced AQP2 protein abundance was significantly attenuated without changes in AQP2 mRNA expression. Moreover, the AQP2 protein abundance was markedly declined during the dDAVP withdrawal period after stimulation under SNX27 knockdown, which was inhibited by lysosome inhibitors. Autophagy was induced after SNX27 knockdown in mpkCCD cells. Lithium-induced nephrogenic diabetes insipidus in rats revealed a significant downregulation of SNX27 in the kidney inner medulla. Taken together, the PDZ domain-containing SNX27 interacts with AQP2 and depletion of SNX27 contributes to the autophagy-lysosomal degradation of AQP2.


Horizontal Resorption of Fresh-Frozen Corticocancellous Bone Blocks in the Reconstruction of the Atrophic Maxilla at 5 Months.
Reliable implant-supported rehabilitation of an alveolar ridge needs sufficient volume of bone. In order to achieve a prosthetic-driven positioning, bone graft techniques may be required. This prospective cohort study aims to clinically evaluate the amount of resorption of corticocancellous fresh-frozen allografts bone blocks used in the reconstruction of the severe atrophic maxilla. Twenty-two partial and totally edentulous patients underwent bone augmentation procedures with fresh-frozen allogenous blocks from the iliac crest under local anesthesia. Implants were inserted into the grafted sites after a healing period of 5 months. Final fixed prosthesis was delivered ± 4 months later. Ridge width analysis and measurements were performed with a caliper before and after grafting and at implant insertion. Bone biopsies were performed in 16 patients. A total of 98 onlay block allografts were used in 22 patients with an initial mean alveolar ridge width of 3.41 ± 1.36 mm. Early exposure of blocks was observed in four situations and one of these completely resorbed. Mean horizontal bone gain was 3.63 ± 1.28 mm (p < .01). Mean buccal bone resorption between allograph placement and the reopening stage was 0.49 ± 0.54 mm, meaning approximately 7.1% (95% confidence interval: [5.6%, 8.6%]) of total ridge width loss during the integration period. One hundred thirty dental implants were placed with good primary stability (≥ 30 Ncm). Four implants presented early failure before the prosthetic delivery (96.7% implant survival). All patients were successfully rehabilitated. Histomorphometric analysis revealed 20.9 ± 5.8% of vital bone in close contact to the remaining grafted bone. A positive strong correlation (adjusted R(2) = 0.44, p = .003) was found between healing time and vital bone percentage. Augmentation procedures performed using fresh-frozen allografts from the iliac crest are a suitable alternative in the reconstruction of the atrophic maxilla with low resorption rate at 5 months, allowing proper stability of dental implants followed by fixed prosthetic rehabilitation.


Structural and biochemical consequences of pyridoxine-dependent epilepsy mutations that target the aldehyde binding site of aldehyde dehydrogenase ALDH7A1.
In humans, certain mutations in the gene encoding aldehyde dehydrogenase 7A1 are associated with pyridoxine-dependent epilepsy (PDE). Understanding the impact of PDE-causing mutations on the structure and activity of ALDH7A1 could allow for the prediction of symptom-severity and aid the development of patient-specific medical treatments. Herein, we investigate the biochemical and structural consequences of PDE missense mutations targeting residues in the aldehyde substrate binding site: N167S, P169S, A171V, G174V, and W175G. All but G174V could be purified for biochemical and X-ray crystallographic analysis. W175G has a relatively mild kinetic defect, exhibiting a fivefold decrease in kcat with no change in Km . P169S and N167S have moderate defects, characterized by catalytic efficiencies of 20- and 100-times lower than wild-type, respectively. A171V has a profound functional defect, with catalytic efficiency 2000-times lower than wild-type. The crystal structures of the variants are the first for any PDE-associated mutant of ALDH7A1. The structures show that missense mutations that decrease the steric bulk of the side chain tend to create a cavity in the active site. The protein responds by relaxing into the vacant space, and this structural perturbation appears to cause misalignment of the aldehyde substrate in W175G and N167S. The P169S structure is nearly identical to that of the wild-type enzyme; however, analysis of B-factors suggests the catalytic defect may result from altered protein dynamics. The A171V structure suggests that the potential for steric clash with Val171 prevents Glu121 from ion pairing with the amino group of the aldehyde substrate. ENZYMES: Aldehyde dehydrogenase 7A1 (EC184.108.40.206). DATABASES: Coordinates have been deposited in the Protein Data Bank under the following accession codes: 6O4B, 6O4C, 6O4D, 6O4E, 6O4F, 6O4G, 6O4H.


Neurotransmitter properties of spinal interneurons in embryonic and larval zebrafish.
Many classes of spinal interneurons in zebrafish have been described based on morphology, but their transmitter phenotypes are largely unknown. Here we combine back-filling or genetic labeling of spinal interneurons with in situ staining for markers of neurotransmitter phenotypes, including the vesicular glutamate transporter (VGLUT) genes for glutamatergic neurons, the neuronal glycine transporter (GLYT2) for glycinergic neurons, and glutamic acid decarboxylase (GAD) for GABAergic neurons. Neurons positive for VGLUT include the commissural CoPA, MCoD, UCoD, and some of the CoSA neurons. The CiD interneurons, which have ipsilateral descending axons, were also VGLUT-positive, as were the ventrally located VeMe interneurons, whose descending axonal trajectory has not been clearly revealed. Cells positive for GLYT2 include the commissural CoLAs as well as some of the CoBL and CoSA neurons. The CiA cells were the only GLYT2-positive cells with an ipsilateral axon. Cells staining for GAD included, most notably, the dorsal longitudinal ascending (DoLA) and KA interneurons. Our approach allowed us to define the likely transmitter phenotypes of most of the known classes of spinal interneurons. These data provide a foundation for understanding the functional organization of the spinal networks in zebrafish.


Sensitivity analysis approaches applied to systems biology models.
With the rising application of systems biology, sensitivity analysis methods have been widely applied to study the biological systems, including metabolic networks, signalling pathways and genetic circuits. Sensitivity analysis can provide valuable insights about how robust the biological responses are with respect to the changes of biological parameters and which model inputs are the key factors that affect the model outputs. In addition, sensitivity analysis is valuable for guiding experimental analysis, model reduction and parameter estimation. Local and global sensitivity analysis approaches are the two types of sensitivity analysis that are commonly applied in systems biology. Local sensitivity analysis is a classic method that studies the impact of small perturbations on the model outputs. On the other hand, global sensitivity analysis approaches have been applied to understand how the model outputs are affected by large variations of the model input parameters. In this review, the author introduces the basic concepts of sensitivity analysis approaches applied to systems biology models. Moreover, the author discusses the advantages and disadvantages of different sensitivity analysis methods, how to choose a proper sensitivity analysis approach, the available sensitivity analysis tools for systems biology models and the caveats in the interpretation of sensitivity analysis results.


Laser-radiation scattering by cement in the process of hydration: simulation of the dynamics and experiment.
This paper discusses simulation of speckle-field dynamics during coherent light scattering by a cement surface in the process of hydration. Cement particles are represented by the spheres whose sizes and reflection indices are changing during the hydration process. The study of intensity fluctuations of scattered coherent radiation is a suitable technique for the analysis of both fast and slow processes of mineral binder hydration and formation of polycrystalline structures in the process of hardening. The results of simulation are in good agreement with the experimental data.


Comprehensive analysis of the major histocompatibility complex in systemic sclerosis identifies differential HLA associations by clinical and serological subtypes.
The greatest genetic effect reported for systemic sclerosis (SSc) lies in the major histocompatibility complex (MHC) locus. Leveraging the largest SSc genome-wide association study, we aimed to fine-map this region to identify novel human leucocyte antigen (HLA) genetic variants associated with SSc susceptibility and its main clinical and serological subtypes. 9095 patients with SSc and 17 584 controls genome-wide genotyped were used to impute and test single-nucleotide polymorphisms (SNPs) across the MHC, classical HLA alleles and their composite amino acid residues. Additionally, patients were stratified according to their clinical and serological status, namely, limited cutaneous systemic sclerosis (lcSSc), diffuse cutaneous systemic sclerosis (dcSSc), anticentromere (ACA), antitopoisomerase (ATA) and anti-RNApolIII autoantibodies (ARA). Sequential conditional analyses showed nine SNPs, nine classical alleles and seven amino acids that modelled the observed associations with SSc. This confirmed previously reported associations with HLA-DRB1*11:04 and HLA-DPB1*13:01, and revealed a novel association of HLA-B*08:01. Stratified analyses showed specific associations of HLA-DQA1*02:01 with lcSSc, and an exclusive association of HLA-DQA1*05:01 with dcSSc. Similarly, private associations were detected in HLA-DRB1*08:01 and confirmed the previously reported association of HLA-DRB1*07:01 with ACA-positive patients, as opposed to the HLA-DPA1*02:01 and HLA-DQB1*03:01 alleles associated with ATA presentation. This study confirms the contribution of HLA class II and reveals a novel association of HLA class I with SSc, suggesting novel pathways of disease pathogenesis. Furthermore, we describe specific HLA associations with SSc clinical and serological subtypes that could serve as biomarkers of disease severity and progression.


A morphologist's approach to the vagina.
The author reviews basic facts on vaginal histology and ultrastructure with respect to passage of different substances and drugs through the epithelium. Particular interest is given to the action mechanism of topically applied intravaginal estrogen, its binding to receptors in different vaginal cell types, and possible local metabolic routes for estrogen. Differences in estrogen sensitivity between uterus and vagina are discussed as well as the background for the high sensitivity in the vagina.


Immediate effects of high-speed cycling intervals on bradykinesia in Parkinson's disease.
There is growing evidence that speed-based exercise training benefits people with Parkinson's disease (PD). The present study investigates the effects of a single session of volitional, high-speed cycling intervals on a battery of timed functional tests selected for their relevance to the symptom of bradykinesia. Ten subjects with PD (Hoehn-Yahr stage ≤ 3.0) participated in a familiarization session and three test sessions. Functional testing occurred before and after 30 minute sessions in which subjects performed no exercise (NO), pedaled at their preferred cadence (PC), or performed 20, 15-second intervals of high-speed low-resistance cycling (HS-LR). In addition to testing the exercise effects in a within-subjects design, we provide test-retest reliability data, minimal detectable change scores, and correlations among the selected functional tests. Despite the relatively low dose of speed-based exercise, HS-LR elicited significant (p < 0.05) improvements in the four square step test and 10 m walk test. Excepting reaction times, there was high reliability and adequate sensitivity to detect moderate and small differences. Strong correlations among tests of mobility inform the future selection of measures in the experimental design. In addition to what is known about continuous exercise sessions involving high-speed exercise, the present results suggest that brief intervals of HS-LR bicycling are promising and should be examined in a longer duration exercise program.


[Lernaea cyprinacea (Copepoda, Lernaeidae) from crucian carp Carassius carassius from the Dlinnoje Lake in the middle stream of the Vychegda River].
Till 1996 the Vychegda River's flood lakes at the territory of the biological station of the Syktyvkar University were overflowed every spring by water so that fishing took place from time to time. In 1996 a low flood level and intensive fishing resulted in the change of the infection rate of crucians by Lernaea, as well as in the change of the size and sex ratios of the crucian population in the Dlinnoje Lake. Thus, the size pattern of the crucian population became simpler, and it led to the changes in parasite-host relation. As a result, the intensity of the invasion of crucians by Lernaea cyprinacea have increased and the distribution of the parasite in the host population does not fit by the negative-binomial distribution. Crucian females and males play an equal role in the maintenance of the parasite population. In 1984 crucian females of medium and large body length played the main role in that process. Functions of other host groups (fishes of small body length, males) were different. Up to 2001 a significance of "k"-exponent of the negative-binomial distribution, describing the number of copepods in the mature part of crucian population, changed with a small range that suggests a stability of parasite-host relationship in the investigated crucian groups. We suppose that after a period of an unstable state in 1997-2000 the population of the parasite reached a new stable state.


Expression and regulation of ANP receptor subtypes in rat renal glomeruli and papillae.
The expression and regulation of atrial natriuretic peptide (ANP) receptor subtypes were examined in rat renal glomeruli and papillae. In glomeruli, approximately two-thirds of ANP binding sites represented guanylate cyclase-uncoupled ANP clearance receptors (ANPc) with a molecular mass of 64 kDa under reducing conditions. The remainder of glomerular ANP binding sites represented guanylate cyclase-coupled ANP receptors (ANPGC) with a molecular mass of 130 kDa. In rat papillae, only the 130-kDa ANPGC was expressed. In rats adapted to a low-salt diet, saline loading or acute ANP infusion resulted in a decrease in ANPC density, a difference that was not detected when glomeruli were first acidwashed to remove endogenous ANP, indicating that apparent regulation of ANPC reflected prior occupancy by endogenous ANP. Densities of glomerular ANPC and ANPGC were similar in spontaneously hypertensive rats (SHR) compared with those of the Wistar-Kyoto (WKY) controls. However, elimination of prior receptor occupancy revealed a significantly greater expression of glomerular ANPC in SHR compared with WKY rats, without significant differences in the density of the glomerular or papillary ANPGC subtype. The failure of the ANPGC subtype to be regulated may account for our previously reported findings that dietary salt intake does not affect glomerular ANP-stimulated guanosine-3',5'-cyclic monophosphate accumulation despite apparent regulation of ANP receptor density. Whether the increased expression of the ANPC subtype in SHR represents a primary defect or results from induction of ANP clearance receptor expression remains to be determined.


Association of vision loss in glaucoma and age-related macular degeneration with IADL disability.
To determine if glaucoma and/or age-related macular degeneration (AMD) are associated with disability in instrumental activities of daily living (IADLs). Glaucoma subjects (n = 84) with bilateral visual field (VF) loss and AMD subjects (n = 47) with bilateral or severe unilateral visual acuity (VA) loss were compared with 60 subjects with normal vision (controls). Subjects completed a standard IADL disability questionnaire, with disability defined as an inability to perform one or more IADLs unassisted. Disability in one or more IADLs was present in 18.3% of controls as compared with 25.0% of glaucoma subjects (P = 0.34) and 44.7% of AMD subjects (P = 0.003). The specific IADL disabilities occurring more frequently in both AMD and glaucoma subjects were preparing meals, grocery shopping, and out-of-home travelling (P < 0.05 for both). In multivariate logistic regression models run adjusting for age, sex, mental status, comorbidity, and years of education, AMD (odds ratio [OR] = 3.4, P = 0.02) but not glaucoma (OR = 1.4, P = 0.45) was associated with IADL disability. However, among glaucoma and control patients, the odds of IADL disability increased 1.6-fold with every 5 dB of VF loss in the better-seeing eye (P = 0.001). Additionally, severe glaucoma subjects (better-eye MD worse than -13.5 dB) had higher odds of IADL disability (OR = 4.2, P = 0.02). Among AMD and control subjects, every Early Treatment of Diabetic Retinopathy Study line of worse acuity was associated with a greater likelihood of IADL disability (OR = 1.3). VA loss in AMD and severe VF loss in glaucoma are associated with self-reported difficulties with IADLs. These limitations become more likely with increasing magnitude of VA or VF loss.


Investigations of pharmacologic properties of the renal CLC-K1 chloride channel co-expressed with barttin by the use of 2-(p-Chlorophenoxy)propionic acid derivatives and other structurally unrelated chloride channels blockers.
CLC-K chloride channels are expressed in the kidney, where they play a pivotal role in the mechanisms of urine concentration and Na(+) reabsorption. The identification of barttin as an essential beta-subunit of CLC-K channels allowed performance of a pharmacologic characterization of wild-type CLC-K1 expressed in Xenopus oocytes. To this end, a series of 2-(p-chlorophenoxy)propionic acid (CPP) derivatives were screened using the two-microelectrode voltage-clamp technique. Several chemical modifications regarding the phenoxy group of the side chain (elimination of the oxygen atom or of methylenic groups, substitutions of the chlorine atom) did not alter the drug blocking activity, with five different derivatives showing a similar potency. Among these, a derivative of CPP carrying a benzyl group on the chiral center in the place of the methyl group represented the minimal structure for blocking CLC-K1. It inhibited the channel from the extracellular side with an affinity in the 150 micro M range. The blocking potency of this compound is fourfold increased by lowering the extracellular chloride concentration, suggesting that the drug interacts with the channel pore. Concomitantly, the effect of some "classical" Cl(-) channel blockers (9-anthracenecarboxylic acid, 2-(phenylamino)benzoic acid, iminodibenzoic acid, niflumic acid, 5-nitro-2-(3-phenylpropylamino)benzoic acid, 4,4'-diisothiocyanato-2,2'-stilbenedisulfonic acid disodium salt, and 4-acetamido-4'-isothiocyanato-2,2'-stilbenedisulfonic acid disodium salt) was screened. 4,4'-Diisothiocyanato-2,2'-stilbenedisulfonic acid disodium salt was the only one capable of blocking CLC-K1 with a potency similar to the CPP derivative, although in an irreversible manner. The newly identified substances provide a useful tool to investigate the biophysical and physiologic role of these renal channels and a starting point for the development of therapeutic drugs with diuretic action.


A distinct metabolic response characterizes sensitivity to EZH2 inhibition in multiple myeloma.
Multiple myeloma (MM) is a heterogeneous haematological disease that remains clinically challenging. Increased activity of the epigenetic silencer EZH2 is a common feature in patients with poor prognosis. Previous findings have demonstrated that metabolic profiles can be sensitive markers for response to treatment in cancer. While EZH2 inhibition (EZH2i) has proven efficient in inducing cell death in a number of human MM cell lines, we hereby identified a subset of cell lines that despite a global loss of H3K27me3, remains viable after EZH2i. By coupling liquid chromatography-mass spectrometry with gene and miRNA expression profiling, we found that sensitivity to EZH2i correlated with distinct metabolic signatures resulting from a dysregulation of genes involved in methionine cycling. Specifically, EZH2i resulted in a miRNA-mediated downregulation of methionine cycling-associated genes in responsive cells. This induced metabolite accumulation and DNA damage, leading to G2 arrest and apoptosis. Altogether, we unveiled that sensitivity to EZH2i in human MM cell lines is associated with a specific metabolic and gene expression profile post-treatment.


Early experience with targeted therapy as a first-line adjuvant treatment for pediatric low-grade glioma.
Pediatric low-grade gliomas (pLGGs) frequently exhibit dysregulation of the mitogen-activated protein kinase (MAPK) pathway. Targeted therapies, including mutant BRAF inhibitors (dabrafenib) and MEK inhibitors (trametinib), have shown promise in patients in whom conventional chemotherapy has failed. However, few studies have investigated the use of targeted therapy as a first-line treatment for pLGG. Here, the authors reviewed their institutional experience with using a personalized medicine approach to patients with newly diagnosed pLGGs. All pediatric patients at the authors' institution who had been treated with dabrafenib or trametinib for pLGG without first receiving conventional chemotherapy or radiation were retrospectively reviewed. Demographic, clinical, and radiological data were collected. Eight patients underwent targeted therapy as a first-line treatment for pLGG. Five patients had a BRAF alteration (1 with a BRAFV600E mutation, 4 with a KIAA1549:BRAF fusion), and 3 patients had an NF1 mutation. One of the 8 patients was initially treated with dabrafenib, and trametinib was added later. Seven patients were initially treated with trametinib; of these, 2 later transitioned to dual therapy, whereas 5 continued with trametinib monotherapy. Six patients (75%) demonstrated a partial response to therapy during their treatment course, whereas stable disease was identified in the remaining 2 patients (25%). One patient experienced mild disease progression after completing a course of trametinib monotherapy, but ultimately stabilized after a period of close observation. Another patient experienced tumor progression while on dabrafenib, but subsequently responded to dual therapy with dabrafenib and trametinib. The most common adverse reactions to targeted therapy were cutaneous toxicity (100%) and diarrhea (50%). Targeted therapies have the potential to become a standard treatment option for pLGG due to their favorable toxicity profile and oral route of administration. This case series provides preliminary evidence that targeted therapies can induce an early disease response as a first-line adjuvant treatment; however, large-scale studies are required to assess long-term durability and safety.


Migration of human mesenchymal stem cells under low shear stress mediated by mitogen-activated protein kinase signaling.
Human mesenchymal stem cells (hMSCs) are attractive candidates for cell-based tissue repair approaches and have been used as vectors for delivering therapeutic genes to sites of injury. It is believed that hMSCs are able to detect and respond to shear stress due to blood and interstitial fluid flow through mechanotransduction pathways after transplantation. However, information regarding hMSC migration under shear stress and its mechanism is still limited. In this study, we examined the effect of shear stress on hMSC migration and the role of mitogen-activated protein kinases (MAPKs) in their migration. Shear stress between 0.2 and 10 Pa, which was produced by the flow medium, was exerted on fluorescently labeled hMSCs. Cell migration was evaluated using the scratch wound assay, and images were captured using a microscope equipped with a digital 3CCD camera. The results showed that hMSCs subjected to a shear stress of 0.2 Pa caused notably faster wound closure than statically cultured hMSCs, while migration in the 0.5- and 1-Pa shear stress group did not differ significantly from that in the control group. Shear stress >2 Pa markedly inhibited hMSC migration. hMSCs subjected to a shear stress of 0.2 Pa displayed an increase in extracellular signal-regulated kinases 1/2 (ERK1/2), c-Jun N-terminal kinases (JNK), and p38 MAPK activation for up to 60 min, while a shear stress of 2 Pa abrogated the activation. JNK and p38 MAPK inhibitors completely abolished the effect of shear stress on hMSC migration, while significant differences were observed between the ERK1/2 inhibitor-treated static control and shear stress groups. Taken together, these results demonstrate that low shear stress effectively induces hMSC migration and that JNK and p38 MAPK play more prominent roles in shear stress-induced migration than ERK1/2.


Preparation of submicroscopic spider threads for particle studies.
Many small spiders produce threads of different diameters, some of which are submicroscopic. When the threads are subjected to an environment of tobacco smoke and then examined microscopically, the invisible threads are manifested as particles in a line with no visible connection. These threads are used to support microscopic water droplets in single-particle scattering experiments.


COVID-19, Social Isolation, and Mental Health Among Older Adults: A Digital Catch-22.
One of the most at-risk groups during the COVID-19 crisis is older adults, especially those who live in congregate living settings and seniors' care facilities, are immune-compromised, and/or have other underlying illnesses. Measures undertaken to contain the spread of the virus are far-reaching, and older adults were among the first groups to experience restrictions on face-to-face contact. Although reducing viral transmission is critical, physical distancing is associated with negative psychosocial implications, such as increased rates of depression and anxiety. Promising evidence suggests that participatory digital co-design, defined as the combination of user-centered design and community engagement models, is associated with increased levels of engagement with mobile technologies among individuals with mental health conditions. The COVID-19 pandemic has highlighted shortcomings of existing technologies and challenges in their uptake and usage; however, strategies such as co-design may be leveraged to address these challenges both in the adaptation of existing technologies and the development of new technologies. By incorporating these strategies, it is hoped that we can offset some of the negative mental health implications for older adults in the context of physical distancing both during and beyond the current pandemic.


Beyond the Global Brain Differences: Intraindividual Variability Differences in 1q21.1 Distal and 15q11.2 BP1-BP2 Deletion Carriers.
Carriers of the 1q21.1 distal and 15q11.2 BP1-BP2 copy number variants exhibit regional and global brain differences compared with noncarriers. However, interpreting regional differences is challenging if a global difference drives the regional brain differences. Intraindividual variability measures can be used to test for regional differences beyond global differences in brain structure. Magnetic resonance imaging data were used to obtain regional brain values for 1q21.1 distal deletion (n = 30) and duplication (n = 27) and 15q11.2 BP1-BP2 deletion (n = 170) and duplication (n = 243) carriers and matched noncarriers (n = 2350). Regional intra-deviation scores, i.e., the standardized difference between an individual's regional difference and global difference, were used to test for regional differences that diverge from the global difference. For the 1q21.1 distal deletion carriers, cortical surface area for regions in the medial visual cortex, posterior cingulate, and temporal pole differed less and regions in the prefrontal and superior temporal cortex differed more than the global difference in cortical surface area. For the 15q11.2 BP1-BP2 deletion carriers, cortical thickness in regions in the medial visual cortex, auditory cortex, and temporal pole differed less and the prefrontal and somatosensory cortex differed more than the global difference in cortical thickness. We find evidence for regional effects beyond differences in global brain measures in 1q21.1 distal and 15q11.2 BP1-BP2 copy number variants. The results provide new insight into brain profiling of the 1q21.1 distal and 15q11.2 BP1-BP2 copy number variants, with the potential to increase understanding of the mechanisms involved in altered neurodevelopment.


Characterization of cytochrome b diversity in Chinese domestic horses.
Previous mitochondrial DNA (mtDNA) D-loop and microsatellite studies have shown that Chinese horses have multiple maternal origins and high genetic diversity. To better characterize maternal genetic origins and diversity of Chinese domestic horses, we conducted a comprehensive analysis of 407 complete 1140 bp sequences of the horse mitochondrially encoded cytochrome b (CYTB) gene, including 323 horses from 13 Chinese indigenous breeds and 84 reference sequences from GenBank. A total of 114 haplotypes were identified, of which 73 appeared among the 13 Chinese horse breeds. The high mitochondrially encoded cytochrome b haplotypic diversity suggests multiple maternal origins in Chinese horses.


Immediate non-occlusal loading of immediate post-extractive versus delayed placement of single implants in preserved sockets of the anterior maxilla: 4-month post-loading results from a pragmatic multicentre randomised controlled trial.
To compare the effectiveness of immediate post-extractive single implants with delayed implants placed in preserved sockets after 4 months of healing. Implants that achieved an insertion torque of at least 35 Ncm were immediately non-occlusally loaded. Just after tooth extraction and in the presence of a loss of the buccal plate bone less than 4 mm, compared to the palatal wall, 106 patients requiring a single immediate postextractive implant in the maxilla from second premolar to second premolar were randomly allocated to immediate implant placement (immediate group; 54 patients) or to socket preservation using anorganic bovine bone covered by a resorbable collagen barrier (delayed group; 52 patients) according to a parallel group design at three different centres. Bone-to-implant gaps were to be filled with anorganic bovine bone, however this was not done in 17 patients (corresponding to 40% of those who should have been grafted). Four months after socket preservation, delayed implants were placed. Implants placed with an insertion torque >35 Ncm were immediately loaded with non-occluding provisional single crowns, replaced, after 4 months, by definitive crowns. Outcome measures were implant failures, complications, aesthetics assessed using the pink esthetic score (PES), and patient satisfaction, recorded by blinded assessors. All patients were followed up to 4 months after loading. Nineteen (35%) implants were not immediately loaded in the immediate group versus 39 (75%) implants in the delayed placement group because an insertion torque >35 Ncm could not be obtained. No patient dropped out. Two implants failed in the immediate group (4%) versus none in the delayed group. More minor complications occurred in the immediate group (8) than the in the delayed group (1) and this was statistically significant (P = 0.032). At delivery of definitive crowns, 4 months after loading, aesthetics were scored as 12.8 and 12.6 in the immediate and delayed groups, respectively. There was no statistically significant difference (P = 0.5). Patients of both groups were equally satisfied. There were more complications at immediate post-extractive implants when compared to delayed implants. The aesthetic outcome appears to be similar for both groups and it seems more difficult to obtain a high insertion torque in sockets preserved with anorganic bovine bone.


The early course and treatment of posttraumatic stress disorder in very young children: diagnostic prevalence and predictors in hospital-attending children and a randomized controlled proof-of-concept trial of trauma-focused cognitive therapy, for 3- to 8-year-olds.
The introduction of developmentally adapted criteria for posttraumatic stress disorder (PTSD) has improved the identification of ≤6-year-old children with clinical needs. Across two studies, we assess predictors of the development of PTSD in young children (PTSD-YC), including the adult-led acute stress disorder (ASD) diagnosis, and provide proof of principle for cognitive-focused therapy for this age range, with the aim of increasing treatment options for children diagnosed with PTSD-YC. Study 1 (N = 105) assessed ASD and PTSD-YC diagnosis in 3- to 8-year-old children within one month and at around three months following attendance at an emergency room. Study 2 (N = 37) was a preregistered (www.isrctn.com/ISRCTN35018680) randomized controlled early-phase trial comparing CBT-3M, a cognitive-focused intervention, to treatment-as-usual (TAU) delivered within the UK NHS to 3- to 8-year-olds diagnosed with PTSD-YC. In Study 1, the ASD diagnosis failed to identify any young children. In contrast, prevalence of acute PTSD-YC (minus the duration requirement) was 8.6% in the first month post-trauma and 10.1% at 3 months. Length of hospital stay, but no other demographic or trauma-related characteristics, predicted development of later PTSD-YC. Early (within one month) diagnosis of acute PTSD-YC had a positive predictive value of 50% for later PTSD-YC. In Study 2, most children lost their PTSD-YC diagnosis following completion of CBT-3M (84.6%) relative to TAU (6.7%) and CBT-3M was acceptable to recipient families. Effect sizes were also in favor of CBT-3M for secondary outcome measures. The ASD diagnosis is not fit for purpose in this age-group. There was a strong and encouraging signal of putative efficacy for young children treated using a cognitive-focused treatment for PTSD, and a larger trial of CBT-3M is now warranted.


Cytokine- and interferon-modulating properties of Echinacea spp. root tinctures stored at -20 degrees C for 2 years.
Echinacea spp. phytomedicines are popular for treating upper respiratory infections. The purpose of this investigation was to examine the immunomodulatory properties of Echinacea tinctures from seven species after being stored at -20 degrees C for 2 years. Two experimental techniques were employed using human peripheral blood mononuclear cells (PBMC). In the first set of experiments, PBMCs were stimulated in vitro with tinctures alone and assayed for proliferation and production of interleukin-10 (IL-10), IL-12, and tumor necrosis factor-alpha (TNF-alpha). In the second set of experiments, subjects were immunized with influenza vaccine. PBMCs from vaccinated individuals were stimulated in vitro with Echinacea tinctures and influenza virus; cytokine production (IL-2, IL-10, and interferon-gamma [IFN-gamma]) was compared prevaccination and postvaccination. In the first experiments, (1) tinctures from E. angustifolia, E. pallida, E. paradoxa, and E. tennesseensis stimulated proliferation and tended to increase IL-10, (2) E. sanguinea and E. simulata stimulated only proliferation, (3) E. purpurea stimulated only IL-10, and (4) none of the extracts influenced IL-12 or TNF-alpha. In the second experiments, (1) tinctures from E. pallida, E. paradoxa, E. sanguinea, and E. simulata diminished influenza-specific IL-2, and (2) none of the extracts influenced influenza-specific IL-10 or IFN-gamma. For in vitro models using Echinacea, immune response may vary based on stimulus (Echinacea alone vs. Echinacea + recall stimulation with virus).


Wagner's canalization model.
Wagner (1996, Does evolutionary plasticity evolve? Evolution 50, 1008-1023.) and Siegal and Bergman, 2002 and Azevedo et al., 2006 have studied a simple model of the evolution of a network of N genes, in order to explain the observed phenomenon that systems evolve to be robust. These authors primarily considered the case N=10 and used simulations to reach their conclusions. Here we investigate this model in more detail, considering systems of different sizes with and without recombination, and with selection for convergence instead of to a specified limit. For the simpler evolutionary model lacking recombination, we analyze the system as a neutral network. This allows us to describe the equilibrium distribution networks within genotype space. Our results show that, given a sufficiently large population size, the qualitative observation that systems evolve to be robust, is itself robust, as it does not depend on the details of the model. In simple terms, robust systems have more viable offspring, so the evolution of robustness is merely selection for increased fecundity, an observation that is well known in the theory of neutral networks.


Site-directed mutagenesis of the peripheral benzodiazepine receptor: identification of amino acids implicated in the binding site of Ro5-4864.
The peripheral benzodiazepine receptor (PBR) is an 18-kDa protein present in the outer mitochondrial membrane. The human PBR can be labeled with the benzodiazepine Ro5-4864 and with the isoquinoline carboxamide PK11195. The two ligands compete with each other in binding experiments, with previous results suggesting overlapping but not identical binding sites. To define the regions of the receptor interacting with PK11195 and Ro5-4864 and to address the question of the topology of the molecule in the membrane, we generated mutant human PBRs with amino- and carboxyl-terminal deletions and with point mutations in potentially accessible cytoplasmic regions. The mutant genes were expressed in yeast and analyzed in binding experiments using radiolabeled PK11195 and Ro5-4864. The results showed that, whereas deletions in the amino-terminal sequence had marked consequences for the binding affinity of both ligands, the final 13 amino acids at the carboxyl terminus could be deleted with no effect on the binding of either Ro5-4864 or PK11195. The site-directed mutagenesis experiments pinpointed four amino acids as participating in the binding site of Ro5-4864. Three of these, Glu-29, Arg-32, and Lys-39, which are located in the first putative cytoplasmic loop, are conserved in human, bovine, rat, and mouse PBRs. The remaining residue, Val-154, which is found at the interface between the putative fifth transmembrane region and the cytoplasm, is present in the human, rat, and mouse sequences but is replaced by methionine in the bovine sequence. The exchange of Met-154 for valine in the bovine PBR introduced a binding site for Ro5-4864, which is absent in the native PBR. These four amino acids played a minor role, if any, in the binding site of PK11195. We also showed that the histidines previously suggested to be part of the binding site of PK11195 are not directly involved in the interaction of the human receptor with either PK11195 or Ro5-4864.


Changing pattern of residual tumor in Burkitt lymphoma: findings at autopsy.
Autopsies were performed on 47 patients who died of Burkitt lymphoma in Uganda between 1967 and 1971. The patients were separated into four groups based on the duration of survival from the time of diagnosis. Twenty patients survived less than three weeks; all had massive tumors. Six patients died between 10 and 16 weeks after diagnosis, usually from side effects of drug therapy. Of 16 patients who died between 24 and 52 weeks after diagnosis, 13 showed involvement of the central nervous system. Five patients died one year or more after diagnosis; two had a late relapse following a disease-free interval of one and four years, respectively; the other three had survived with clinically apparent tumor that was not responsive to therapy. In recent years, there has been a steady reduction in the proportion of patients showing residual systemic involvement at autopsy. This reduction is balanced by an increase in the proportion of patients with lesions of the central nervous system.