The efficacy of the relaxation response in preparing for cardiac surgery.
This study evaluated the efficacy of the relaxation response on the post-operative recovery of 27 cardiac surgery patients randomly assigned to one of two groups. Thirteen experimental group patients received educational information and practiced eliciting the relaxation response before and after surgery. The 14 patients in the control group received only information. Experimental and control groups were compared before and after surgery on both physiological and psychological recovery variables. There were no initial differences between experimental and control groups on demographic, physiological, and most psychological variables. The experimental group had lower incidence of postoperative supraventricular tachycardia (SVT) than the control group (p = .04) despite having had the same occurrence previously. Experimental and control groups did not significantly differ over the course of study on any other physiological variables. Patients practicing the relaxation response had greater decreases in psychological tension (p = .04) and anger (p = .04) than those who received only educational information. The decreases in psychological tension may have been a result of regression to the mean because the experimental group started with elevated tension relative to the control group (p = .04). We conclude that practicing the relaxation response before and after surgery may reduce SVT, tension, and anger.


Elective abdominal hysterectomy in Nigerian Jehovah's Witnesses.
In a retrospective study at a university hospital, the perioperative morbidity associated with elective total abdominal hysterectomy in 23 Jehovah's Witnesses was compared with that of 46 non-Witness controls. The mean operative blood loss was significantly less, the procedure was lengthier and the average postoperative hospital stay was longer in the study than in the control group. Febrile morbidity was insignificantly more frequent among the study group (OR: 2.05, CI: 0.61-6.88) and there was no significant difference between the overall morbidity experienced by patients in both groups (study: 43.5% versus control: 39.1%; P = 0.73). The perioperative morbidity associated with elective abdominal hysterectomy in patients unwilling to accept blood transfusion does not justify the denial of this important gynaecological surgery when indicated. Gynaecologists in poor resource settings should consciously aim at providing 'bloodless' care for all their patients undergoing abdominal hysterectomy as this may translate to reduced blood loss and decreased need for blood transfusion.


Dual-responsive self-assembly in lysosomes enables cell cycle arrest for locking glioma cell growth.
Herein we first report a dual-responsive peptide substrate (Comp. 1) for preparing self-assembled nanomaterials triggered by pH and legumain. The dual-responsive self-assembly of Comp. 1 in glioma cells enables its long retention time in lysosomes, S phase arrest, and cell growth locking. We verified that the blocked degradation of HIF-1α in lysosomes played a key role in cell cycle arrest and decreased DNA replication. This work illustrates the disturbance of lysosomal function by self-assembled nanomaterials as a promising strategy for inhibiting glioma cell growth.


Characterization of Laguncularia racemosa transcriptome and molecular response to oil pollution.
Mangroves are ecosystems of economic and ecological importance. Laguncularia racemosa (Combretaceae), popularly known as white mangrove, is a species that greatly contributes to the community structure of neotropical and West African mangrove forests. Despite the significance of these ecosystems, they have been destroyed by oil spills that can cause yellowing of leaves, increased sensitivity to other stresses and death of trees. However, the molecular response of plants to oil stress is poorly known. In this work, Illumina reads were de novo assembled into 46,944 transcripts of L. racemosa roots and leaves, including putative isoform variants. In addition to improving the genomic information available for mangroves, the L. racemosa assembled transcriptome allowed us to identify reference genes to normalize quantitative real-time PCR (qPCR) expression data from oil-stressed mangrove plants, which were used in RNASeq validation. The analysis of expression changes induced by the oil exposure revealed 310 and 286 responsive transcripts of leaves and roots, respectively, mainly up-regulated. Enriched GO categories related to chloroplasts and photosynthesis were found among both leaf and root oil-responsive transcripts, while "response to heat" and "response to hypoxia" were exclusively enriched in leaves and roots, respectively. The comparison of L. racemosa 12-h-oil-stressed leaf expression profile to previous Arabidopsis heat-stress studies and co-expression evidence also pointed to similarities between the heat and oil responses, in which the HSP-coding genes seem to play a key role. A subset of the L. racemosa oil-responsive root genes exhibited similar up-regulation profiles to their Arabidopsis homologs involved in hypoxia responses, including the HRA1 and LBD41 TF-coding genes. Genes linked to the ethylene pathway such as those coding for ERF TFs were also modulated during the L. racemosa root response to oil stress. Taken together, these results show that oil contamination affects photosynthesis, protein metabolism, hypoxia response and the ethylene pathway in L. racemosa 12-h-oil-exposed leaves and roots.


[Complications of cataract surgery. Retrospective study of 1,304 cases].
This retrospective study concerns complications arising from 1304 first operations for cataract carried out between 1982 and 1987 by two surgeons having followed the same training and using the same techniques. Loss of vitreous (2.5%) cystoid macular edema (1.9%), and retinal detachement (1.1%) are those complications occurring most frequently in this series. Other serious complications, in descending order, are trauma (0.7%), iris prolaps (0.5%), corneal dystrophy (0.4%), pupillary block (0.3%), expulsive hemorrhage (0.15%), loss of the nucleus in the vitreous (0.07%), panophthalmia (0.07%). Secondary cataract was noted in 31% of cases.


Etiology-Specific Mineralization Patterns in Patients with Labyrinthitis Ossificans.
Our aim was to identify whether specific patterns of ossification in labyrinthitis ossificans are associated with the known risk factors. Labyrinthitis ossificans has been described as sequela of prior temporal bone trauma, prior infection, and other disorders including sickle cell disease. Specific patterns of mineralization in the membranous labyrinth associated with these risk factors has not been previously described. This was a retrospective study evaluating temporal bone CT scans at our institution from November 2005 to May 2018 in patients with labyrinthitis ossificans. Membranous labyrinthine structures evaluated for ossification included the following: basal, middle, and apical cochlear turns; lateral, posterior, and superior semicircular canals; and the vestibule for both ears in all patients. These structures were assigned a severity score, 0-4, based on degree of mineralization. Clinical records were reviewed for potential labyrinthitis ossificans risk factors. Basic descriptive statistics and a mixed model were used to correlate the degree and patterns of ossification with clinical history. Forty-four patients (58 ears) with labyrinthitis ossificans were identified and evaluated. The most common risk factors were chronic otomastoiditis (n = 18), temporal bone surgery (n = 9), temporal bone trauma (n = 6), sickle cell disease (n = 5), and meningitis (n = 4). For all etiologies, the semicircular canals were most severely affected, and the vestibule was the least. In patients with prior temporal bone surgery, significantly greater mineralization was seen in the basal turn of the cochlea (P = .027), the vestibule (P = .001), and semicircular canals (P < .001-.008). No significant pattern was identified in patients with meningitis, sickle cell disease, or trauma. Significant patterns of mineralization in labyrinthitis ossificans were observed in patients with prior temporal bone surgery. For all etiologies, the semicircular canals were most severely affected. No significant mineralization pattern was observed in patients with chronic otomastoiditis, meningitis, sickle cell disease, or prior temporal bone trauma.


Downstaging in pancreatic cancer: a matched analysis of patients resected following systemic treatment of initially locally unresectable disease.
Patients with locally unresectable pancreatic cancer (AJCC stage III) have a median survival of 10-14 months. The objective of this study was to evaluate outcome of initially unresectable patients who respond to multimodality therapy and undergo resection. Using a prospectively collected database, patients were identified who were initially unresectable because of vascular invasion and had sufficient response to nonoperative treatment to undergo resection. Overall survival (OS) was compared with a matched group of patients who were initially resectable. Case matching was performed using a previously validated pancreatic cancer nomogram. A total of 36 patients with initial stage III disease were identified who underwent resection after treatment with either systemic therapy or chemoradiation. Initial unresectability was determined by operative exploration (n = 15, 42%) or by cross-sectional imaging (n = 21, 58%). Resection consisted of pancreaticoduodenectomy (n = 31, 86%), distal pancreatectomy (n = 4, 11%), and total pancreatectomy (n = 1, 3%). Pathology revealed T3 lesions in 26 patients (73%), node positivity in 6 patients (16%), and a negative margin in 30 patients (83%). The median OS in this series was 25 months from resection and 30 months since treatment initiation. There was no difference in OS from time of resection between the initial stage III patients and those who presented with resectable disease (P = .35). In this study, patients who were able to undergo resection following treatment of initial stage III pancreatic cancer experienced survival similar to those who were initially resectable. Resection is indicated in this highly select group of patients.


Degradation of tetra- and trichloroethylene under iron reducing conditions by Acidimicrobiaceae sp. A6.
The degradation of trichloroethylene (TCE) and tetrachloroethylene (PCE), in incubations where ammonium was oxidized while iron was being reduced indicates that these compounds can be degraded during the Feammox process by Acidimicrobiaceae sp. A6 (ATCC, PTA-122488). None of these compounds were degraded in incubations to which no ammonium was added, indicating that they were degraded during the oxidation of ammonium. Degradation of TCE and PCE (ranging between 32% and 55%) was observed in incubations with a pure Acidimicrobiaceae sp. A6 culture as well as an Acidimicrobiaceae sp. A6 enrichment culture over a 2-week period. In addition to these batch studies, a column study, with a 5-h hydraulic residence time, was conducted contrasting the degradation of TCE in iron-rich soil columns that were either seeded with a pure or an enrichment culture of Acidimicrobiaceae sp. A6 to achieve ammonium oxidation under iron reduction, and a control column that was initially not seeded and later seeded with Geobacter metallireducens. While there was ∼22% TCE removal in the columns seeded with Acidimicrobiaceae sp. A6, there was no removal in the unseeded column or the column seeded with G. metallireducens which was being operated under iron reducing conditions. Feammox is an anoxic process that requires acidic conditions. Hence, these results indicate that this process might be harnessed where other bioremediation strategies are difficult, since many require neutral or alkaline conditions, and supplying ammonium to an anoxic aquifer is relatively easy, since there are not many processes that will oxidize ammonium in the absence of dissolved oxygen.


Hierarchical scaffolds enhance osteogenic differentiation of human Wharton's jelly derived stem cells.
Hierarchical structures, constituted by polymeric nano and microfibers, have been considered promising scaffolds for tissue engineering strategies, mainly because they mimic, in some way, the complexity and nanoscale detail observed in real organs. The chondrogenic potential of these scaffolds has been previously demonstrated, but their osteogenic potential is not yet corroborated. In order to assess if a hierarchical structure, with nanoscale details incorporated, is an improved scaffold for bone tissue regeneration, we evaluate cell adhesion, proliferation, and osteogenic differentiation of human Wharton's jelly derived stem cells (hWJSCs), seeded into hierarchical fibrous scaffolds. Biological data corroborates that hierarchical fibrous scaffolds show an enhanced cell entrapment when compared to rapid prototyped scaffolds without nanofibers. Furthermore, upregulation of bone specific genes and calcium phosphate deposition confirms the successful osteogenic differentiation of hWJSCs on these scaffolds. These results support our hypothesis that a scaffold with hierarchical structure, in conjugation with hWJSCs, represents a possible feasible strategy for bone tissue engineering applications.


Robust Tests for Additive Gene-Environment Interaction in Case-Control Studies Using Gene-Environment Independence.
There have been recent proposals advocating the use of additive gene-environment interaction instead of the widely used multiplicative scale, as a more relevant public health measure. Using gene-environment independence enhances statistical power for testing multiplicative interaction in case-control studies. However, under departure from this assumption, substantial bias in the estimates and inflated type I error in the corresponding tests can occur. In this paper, we extend the empirical Bayes (EB) approach previously developed for multiplicative interaction, which trades off between bias and efficiency in a data-adaptive way, to the additive scale. An EB estimator of the relative excess risk due to interaction is derived, and the corresponding Wald test is proposed with a general regression setting under a retrospective likelihood framework. We study the impact of gene-environment association on the resultant test with case-control data. Our simulation studies suggest that the EB approach uses the gene-environment independence assumption in a data-adaptive way and provides a gain in power compared with the standard logistic regression analysis and better control of type I error when compared with the analysis assuming gene-environment independence. We illustrate the methods with data from the Ovarian Cancer Association Consortium.


Optimal expression of cloned NMDAR1/NMDAR2A heteromeric glutamate receptors: a biochemical characterization.
The N-methyl-D-aspartate R1 (NMDAR1) and NMDAR2A subunits were expressed transiently either alone or in combination in human embryonic kidney (HEK) 293 cells. The biochemical and pharmacological properties of the cloned receptors were compared with those of adult rat brain NMDA receptors using both immunological methods with a newly developed anti-NMDAR2A-(1435-1445) antibody and [3H]MK801 radioligand binding activity. Anti-NMDAR2A-(1435-1445) antibodies recognized specifically four immunoreactive species with M(r)s of 180,000, 122,000, 97,000 and 54,000 in rat brain, but only a single band of M(r) 180,000 in HEK 293 cells singly transfected with plasmid pCISNMDAR2A. N-deglycosylation of HEK cell membranes yielded a 165,000-M(r) immunoreactive species, which is in agreement with the size predicted from the cDNA sequence for the mature NMDAR2A subunit. Co-expression of NMDAR1 and NMDAR2A subunits in HEK 293 cells resulted in cell death. Thus conditions were established for the optimum expression of heteromeric receptors in viable cells, including a requirement for DL-2-amino-5-phosphonopentanoic acid (AP5) in the culture medium post-transfection. Cells transfected with pCISNMDAR1 and pCISNMDAR2A combined yielded a 10-fold increase in the number of [3H]MK801 binding sites compared with single subunit expression. MK801 had similar affinity for the expressed receptors as for those found in adult rat and mouse brain. These results demonstrate that the NMDAR1 and NMDAR2A receptor subunits co-assemble to form a heteromeric complex with properties similar to those of the native receptors of adult mammalian forebrain. Furthermore, the conditions reported for maximal transient expression provide a basis for further structure-activity studies.


Panoptic Studio: A Massively Multiview System for Social Interaction Capture.
We present an approach to capture the 3D motion of a group of people engaged in a social interaction. The core challenges in capturing social interactions are: (1) occlusion is functional and frequent; (2) subtle motion needs to be measured over a space large enough to host a social group; (3) human appearance and configuration variation is immense; and (4) attaching markers to the body may prime the nature of interactions. The Panoptic Studio is a system organized around the thesis that social interactions should be measured through the integration of perceptual analyses over a large variety of view points. We present a modularized system designed around this principle, consisting of integrated structural, hardware, and software innovations. The system takes, as input, 480 synchronized video streams of multiple people engaged in social activities, and produces, as output, the labeled time-varying 3D structure of anatomical landmarks on individuals in the space. Our algorithm is designed to fuse the "weak" perceptual processes in the large number of views by progressively generating skeletal proposals from low-level appearance cues, and a framework for temporal refinement is also presented by associating body parts to reconstructed dense 3D trajectory stream. Our system and method are the first in reconstructing full body motion of more than five people engaged in social interactions without using markers. We also empirically demonstrate the impact of the number of views in achieving this goal.


Mobile Phone Messaging to Husbands to Improve Maternal and Child Health Behavior in India.
Over 90% of households in rural Uttar Pradesh, the most populous state of India, have at least one mobile phone. However, ownership of mobile phone among women is quite low. Implementation research was conducted in Uttar Pradesh to examine (a) whether providing information on selected maternal and child health (MCH) behaviors to a husband's mobile phone would enhance the man's knowledge and lead to discussions in their family, and (b) whether such discussions would help in the adoption of healthy practices. The m-Health intervention included biweekly voice messages in local language (Hindi) on MCH topics to the mobile phone of pregnant women's husbands. Using a quasi-experimental design, after four months of the m-Health intervention, in 2014, 881 husbands and 956 women from the study area were interviewed. Husbands' knowledge, controlling for their socio-demographic characteristics, were significantly higher among the listeners of the messages than the non-listeners. Multivariate logistic regression analysis showed that if husbands discussed the messages with family members, the odds of wives' practicing health behaviors improved significantly for three behaviors. These include one antenatal checkup in last trimester of pregnancy (odds ratio 1.72, p < 0.05), receiving a postnatal checkup within 7 days of delivery (odds ratio 3.02, p < 0.05), and delayed bathing of newborn (odds ratio 1.93, p < 0.05). Thus, communicating messages using m-Health was found to be an effective intervention for behavior change. The study demonstrated that mobile phones can be used effectively to reach men with MCH information and encourage them to promote healthy behavior in their family.


Use of carbon monoxide in minimizing ischemia/reperfusion injury in transplantation.
Although carbon monoxide (CO) is known to be toxic because of its ability to interfere with oxygen delivery at high concentrations, mammalian cells endogenously generate CO primarily via the catalysis of heme by heme oxygenases. Recent findings have indicated that heme oxygenases and generation of CO serve as a key mechanism to maintain the integrity of the physiological function of organs and supported the development of a new paradigm that CO, at low concentrations, functions as a signaling molecule in the body and exerts significant cytoprotection. Consequently, exogenously delivered CO has been shown to mediate potent protection in various injury models through its anti-inflammatory, vasodilating, and antiapoptotic functions. Ischemia/reperfusion (I/R) injury associated with organ transplantation is one of the major deleterious factors limiting the success of transplantation. Ischemia/reperfusion injury is a complex cascade of interconnected events involving cell damage, apoptosis, vigorous inflammatory responses, microcirculation disturbance, and thrombogenesis. Carbon monoxide has a great potential in minimizing I/R injury. This review will provide an overview of the basic physiology of CO, preclinical studies examining efficacy of CO in I/R injury models, and possible protective mechanisms. Carbon monoxide could be developed to be a valuable therapeutic molecule in minimizing I/R injury in transplantation.


Gut Microbiota-Based Pharmacokinetics and the Antidepressant Mechanism of Paeoniflorin.
Paeoniflorin, the main component of Xiaoyao Wan, presents low oral bioavailability and unclear antidepressant mechanism. To elucidate the potential reasons for the low bioavailability of paeoniflorin and explore its antidepressant mechanism from the perspective of the gut microbiota, here, a chronic unpredictable depression model and forced swimming test were firstly performed to examine the antidepressant effects of paeoniflorin. Then the pharmacokinetic study of paeoniflorin in rats was performed based on the gut microbiota; meanwhile, the gut microbiota incubated with paeoniflorin in vitro was used to identify the possible metabolites of paeoniflorin. Molecular virtual docking experiments together with the specific inhibitor tests were applied to investigate the mechanism of paeoniflorin metabolism by the gut microbiota. Finally, the intestinal microbiota composition was analyzed by 16S rRNA gene sequencing technology. The pharmacodynamics tests showed that paeoniflorin had significant antidepressant activity, but its oral bioavailability was 2.32%. Interestingly, we found paeoniflorin was converted into benzoic acid by the gut microbiota, and was mainly excreted through the urine with the gut metabolite benzoic acid as the prominent excreted form. Moreover, paeoniflorin could also regulate the composition of the gut microbiota by increasing the abundance of probiotics. Therefore, the metabolism effect of gut microbiota may be one of the main reasons for the low oral bioavailability of paeoniflorin. Additionally, paeoniflorin can be metabolized into benzoic acid via gut microbiota enzymes, which might exert antidepressant effects through the blood-brain barrier into the brain.


Eosinophil-mediated killing of Haemonchus contortus larvae: effect of eosinophil activation and role of antibody, complement and interleukin-5.
Eosinophils have previously been shown to accumulate around the tissue invasive (L3) stage of sheep gastrointestinal parasites in vivo. In this study, eosinophils obtained from mammary washes of sheep, were shown to immobilize and kill H. contortus larvae in vitro in the presence of antibody specific against a defined L3 surface antigen. Eosinophils obtained from sheep primed by repeated infusion of H. contortus larvae were more effective than eosinophils obtained after a single infusion of parasite extract in Fasciola hepatica infected ewes suggesting the former were activated in vivo. The level of larval immobilization in the presence of antibody was significantly increased when complement was added to cultures containing activated eosinophils. The addition of interleukin-5 to larval cultures containing antibody and complement resulted in a significant increase in larval immobilization with unactivated eosinophils suggesting that eosinophil effector function is enhanced following priming with this cytokine. Ultrastructural analysis of the eosinophil/larvae interaction at 6 h of incubation revealed degranulation of adhering eosinophils onto the surface of larvae. By 24 h of incubation, many larvae showed signs of damage and most eosinophils had degenerated. These results suggest that eosinophil-mediated killing may be an effector mechanism for the elimination of L3 H. contortus larvae in immune sheep.


Celastrol decreases specificity proteins (Sp) and fibroblast growth factor receptor-3 (FGFR3) in bladder cancer cells.
Celastrol (CSL) is a naturally occurring triterpenoid acid that exhibits anticancer activity, and in KU7 and 253JB-V bladder cells, CSL induced apoptosis, inhibited growth, colony formation and migration and CSL decreased bladder tumor growth in vivo. CSL also decreased expression of specificity protein (Sp) transcription factors Sp1, Sp3 and Sp4 and several Sp-regulated genes/proteins including vascular endothelial growth factor, survivin and cyclin D1 and fibroblast growth factor receptor-3, a potential drug target for bladder cancer therapy, has now been characterized as an Sp-regulated gene downregulated by CSL. The mechanism of Sp downregulation by CSL was cell context-dependent due to activation of proteosome-dependent (KU7) and -independent (253JB-V) pathways. In 253JB-V cells, CSL induced reactive oxygen species (ROS) and inhibitors of ROS blocked CSL-induced growth inhibition and repression of Sp1, Sp3 and Sp4. This response was due to induction of the Sp repressors ZBTB10 and ZBTB4 and downregulation of miR-27a and miR-20a/17-5p, respectively, which regulate expression of these transcriptional repressors. Thus, the anticancer activity of CSL in 253JB-V cells is due to induction of ROS and ROS-mediated induction of Sp repressors (ZBTB4/ZBTB10) through downregulation of miR-27a and miR-20a/17-5p.


Patient comprehension of emergency department care and instructions: are patients aware of when they do not understand?
To be able to adhere to discharge instructions after a visit to the emergency department (ED), patients should understand both the care that they received and their discharge instructions. The objective of this study is to assess, at discharge, patients' comprehension of their ED care and instructions and their awareness of deficiencies in their comprehension. We conducted structured interviews of 140 adult English-speaking patients or their primary caregivers after ED discharge in 2 health systems. Participants rated their subjective understanding of 4 domains: (1) diagnosis and cause; (2) ED care; (3) post-ED care, and (4) return instructions. We assessed patient comprehension as the degree of agreement (concordance) between patients' recall of each of these domains and information obtained from chart review. Two authors scored each case independently and discussed discrepancies before providing a final concordance rating (no concordance, minimal concordance, partial concordance, near concordance, complete concordance). Seventy-eight percent of patients demonstrated deficient comprehension (less than complete concordance) in at least 1 domain; 51% of patients, in 2 or more domains. Greater than a third of these deficiencies (34%) involved patients' understanding of post-ED care, whereas only 15% were for diagnosis and cause. The majority of patients with comprehension deficits failed to perceive them. Patients perceived difficulty with comprehension only 20% of the time when they demonstrated deficient comprehension. Many patients do not understand their ED care or their discharge instructions. Moreover, most patients appear to be unaware of their lack of understanding and report inappropriate confidence in their comprehension and recall.


Multiple Plasticity Regulators Reveal Targets Specifying an Induced Predatory Form in Nematodes.
The ability to translate a single genome into multiple phenotypes, or developmental plasticity, defines how phenotype derives from more than just genes. However, to study the evolutionary targets of plasticity and their evolutionary fates, we need to understand how genetic regulators of plasticity control downstream gene expression. Here, we have identified a transcriptional response specific to polyphenism (i.e., discrete plasticity) in the nematode Pristionchus pacificus. This species produces alternative resource-use morphs-microbivorous and predatory forms, differing in the form of their teeth, a morphological novelty-as influenced by resource availability. Transcriptional profiles common to multiple polyphenism-controlling genes in P. pacificus reveal a suite of environmentally sensitive loci, or ultimate target genes, that make up an induced developmental response. Additionally, in vitro assays show that one polyphenism regulator, the nuclear receptor NHR-40, physically binds to promoters with putative HNF4α (the nuclear receptor class including NHR-40) binding sites, suggesting this receptor may directly regulate genes that describe alternative morphs. Among differentially expressed genes were morph-limited genes, highlighting factors with putative "on-off" function in plasticity regulation. Further, predatory morph-biased genes included candidates-namely, all four P. pacificus homologs of Hsp70, which have HNF4α motifs-whose natural variation in expression matches phenotypic differences among P. pacificus wild isolates. In summary, our study links polyphenism regulatory loci to the transcription producing alternative forms of a morphological novelty. Consequently, our findings establish a platform for determining how specific regulators of morph-biased genes may influence selection on plastic phenotypes.


Phytochemicals as Inhibitors of Candida Biofilm.
Candida biofilm and associated infections is a serious threat to the large population of immunocompromised patients. Biofilm growth on prosthetic devices or host tissue shows reduced sensitivity to antifungal agents and persists as a reservoir of infective cells. Options for successful treatment of biofilm associated Candida infections are restricted because most of the available antifungal drugs fail to eradicate biofilms. Various plant actives are known to possess interesting antifungal properties. To explore and review the potential of phytochemicals as a novel strategy against Candida biofilms is the intent of present article. Thorough literature search is performed to identify Candida biofilm inhibitors of plant origin. An account of efficacy of selected phytochemicals is presented taking into consideration their biofilm inhibitory concentrations. This review discusses biofilm formation by Candida species, their involvement in human infections, and associated drug resistance. It gives insight into the biofilm inhibitory potential of various phytochemicals. Based on the available reports including the work done in our laboratory, several plant extracts, essential oils and phytomolecules have been identified as excellent inhibitors of biofilms of C. albicans and non-albicans Candida species (NACS). Selected phytochemicals which exhibit activities at low concentrations without displaying toxicity to host are potential therapeutic agents against biofilm associated Candida infections. In vivo testing in animal models and clinical trials in humans are required to be taken up seriously to propose few of the phytochemicals as candidate drug molecules.


Cotranscriptional RNA strand exchange underlies the gene regulation mechanism in a purine-sensing transcriptional riboswitch.
RNA folds cotranscriptionally to traverse out-of-equilibrium intermediate structures that are important for RNA function in the context of gene regulation. To investigate this process, here we study the structure and function of the Bacillus subtilis yxjA purine riboswitch, a transcriptional riboswitch that downregulates a nucleoside transporter in response to binding guanine. Although the aptamer and expression platform domain sequences of the yxjA riboswitch do not completely overlap, we hypothesized that a strand exchange process triggers its structural switching in response to ligand binding. In vivo fluorescence assays, structural chemical probing data and experimentally informed secondary structure modeling suggest the presence of a nascent intermediate central helix. The formation of this central helix in the absence of ligand appears to compete with both the aptamer's P1 helix and the expression platform's transcriptional terminator. All-atom molecular dynamics simulations support the hypothesis that ligand binding stabilizes the aptamer P1 helix against central helix strand invasion, thus allowing the terminator to form. These results present a potential model mechanism to explain how ligand binding can induce downstream conformational changes by influencing local strand displacement processes of intermediate folds that could be at play in multiple riboswitch classes.


A Terahertz (THz) Single-Polarization-Single-Mode (SPSM) Photonic Crystal Fiber (PCF).
This paper presents a novel approach to attain a single-polarization-single-mode (SPSM) photonic crystal fiber (PCF) in the terahertz (THz) regime. An initial circular hole PCF design is modified by introducing asymmetry in the first ring of six air holes in the cladding, i.e., epsilon-near-zero (ENZ) material is introduced into only four of those air holes and the other two remain air-filled but have different diameters. The resulting fundamental X-polarized (XP) and Y-polarized (YP) modes have distinctly different electric field distributions. The asymmetry is arranged so that the YP mode has a much larger amount of the field distributed in the ENZ material than the XP mode. Since the ENZ material is very lossy, the YP mode suffers a much higher loss than the XP mode. Consequently, after a short propagation distance, the loss difference (LD) between the XP and YP modes will be large enough that only the XP mode still realistically exists in the PCF. To further enhance the outcome, gain material is introduced into the core area to increase the LDs between the wanted XP mode and any unwanted higher order (HO) modes, as well as to compensate for the XP mode loss without affecting the LD between the XP and YP modes. The optimized PCF exhibits LDs between the desired XP mode and all other modes greater than 8.0 dB/cm across a wide frequency range of 0.312 THz. Consequently, the reported PCF only needs a length of 2.5 cm to attain an SPSM result, with the unwanted modes being more than 20 dB smaller than the wanted mode over the entire operational band.


Immunohistochemical expression and colocalization of somatostatin, carboxypeptidase-E and prohormone convertases 1 and 2 in rat brain.
The processing of many peptides for their maturation in target tissue depends upon the presence of sorting receptor. Several previous studies have predicted that carboxypeptidase-E (CPE), prohormone convertase 1 (PC1) and prohormone convertase 2 (PC2) may function as sorting elements for somatostatin (SST) for its maturation and processing to appropriate targets. However, nothing is currently known about whether brain, neuronal culture or even endocrine cells express SST, CPE, PC1 and PC2 and exhibit colocalization. Accordingly, in the present study using peroxidase immunohistochemistry, double-labeled indirect immunofluorescence immunohistochemistry and Western blot analysis, we mapped the distributional pattern of SST, CPE, PC1 and PC2 in different rat brain regions. Additionally, we also determined the colocalization of SST with CPE, PC1 and PC2 as well as colocalization of CPE with PC1 and PC2. The localization of SST, CPE, PC1 and PC2 reveals a distinct and region specific distribution pattern in the rat brain. Using an indirect double-label immunofluorescence method we observed selective neuron specific colocalization in a region specific manner in cortex, striatum and hippocampus. These studies provide the first evidence for colocalization between SST, CPE, PC1 and PC2 as well as CPE with PC1 and PC2. SST in cerebral cortex colocalized in pyramidal and non-pyramidal neurons with CPE, PC1 and PC2. Most importantly, in striatum and hippocampus colocalization was mostly observed selectively and preferentially in interneurons. CPE is also colocalized with PC1 and PC2 in a region specific manner. The data presented here provide a new insight into the distribution and colocalization of SST, CPE, PC1 and PC2 in rat brain. Taken together, our data anticipate the possibility that CPE, PC1 and PC2 might be potential target for the maturation of SST.


Modeling and simulation in biomedicine.
A group of researchers and educators in The Netherlands, Germany and Czechoslovakia have developed and adapted mathematical computer models of phenomena in the field of physiology and biomedicine for use in higher education. The models are graphical and highly interactive, and are all written in TurboPascal or the mathematical simulation language PSI. An educational shell has been developed to launch the models. The shell allows students to interact with the models and teachers to edit the models, to add new models and to monitor the achievements of the students. The models and the shell have been implemented on a MS-DOS personal computer. This paper describes the features of the modeling package and presents the modeling and simulation of the heart muscle as an example.


Psychosurgery in the modern era: therapeutic and ethical aspects.
Brain operations for psychiatric illness were in the past the centre of public controversy. Fear of untoward side-effects and an exaggerated use of the procedure including treatment of societal aspects of the disease process caused a virtual cessation of psychosurgical operations. With the introduction of techniques that could accurately lesion specific areas in the brain, especially in the limbic system, psychosurgical procedures were revived. Psychiatric syndromes, such as obsessive-compulsive affective disorders, were shown to have benefited from such intervention, without giving rise to untoward side-effects. Given societal supervision it is appropriate to reconsider the use of psychosurgery in well-defined clinical entities of mental disease that do not respond to alternative therapy.


Phase 3 diagnostic evaluation of a smart tablet serious game to identify autism in 760 children 3-5 years old in Sweden and the United Kingdom.
Recent evidence suggests an underlying movement disruption may be a core component of autism spectrum disorder (ASD) and a new, accessible early biomarker. Mobile smart technologies such as iPads contain inertial movement and touch screen sensors capable of recording subsecond movement patterns during gameplay. A previous pilot study employed machine learning analysis of motor patterns recorded from children 3-5 years old. It identified those with ASD from age-matched and gender-matched controls with 93% accuracy, presenting an attractive assessment method suitable for use in the home, clinic or classroom. This is a phase III prospective, diagnostic classification study designed according to the Standards for Reporting Diagnostic Accuracy Studies guidelines. Three cohorts are investigated: children typically developing (TD); children with a clinical diagnosis of ASD and children with a diagnosis of another neurodevelopmental disorder (OND) that is not ASD. The study will be completed in Glasgow, UK and Gothenburg, Sweden. The recruitment target is 760 children (280 TD, 280 ASD and 200 OND). Children play two games on the iPad then a third party data acquisition and analysis algorithm (Play.Care, Harimata) will classify the data as positively or negatively associated with ASD. The results are blind until data collection is complete, when the algorithm's classification will be compared against medical diagnosis. Furthermore, parents of participants in the ASD and OND groups will complete three questionnaires: Strengths and Difficulties Questionnaire; Early Symptomatic Syndromes Eliciting Neurodevelopmental Clinical Examinations Questionnaire and the Adaptive Behavioural Assessment System-3 or Vineland Adaptive Behavior Scales-II. The primary outcome measure is sensitivity and specificity of Play.Care to differentiate ASD children from TD children. Secondary outcomes measures include the accuracy of Play.Care to differentiate ASD children from OND children. This study was approved by the West of Scotland Research Ethics Service Committee 3 and the University of Strathclyde Ethics Committee. Results will be disseminated in peer-reviewed publications and at international scientific conferences. NCT03438994; Pre-results.


Skin langerin+ dendritic cells transport intradermally injected anti-DEC-205 antibodies but are not essential for subsequent cytotoxic CD8+ T cell responses.
Incorporation of Ags by dendritic cells (DCs) increases when Ags are targeted to endocytic receptors by mAbs. We have previously demonstrated in the mouse that mAbs against C-type lectins administered intradermally are taken up by epidermal Langerhans cells (LCs), dermal Langerin(neg) DCs, and dermal Langerin(+) DCs in situ. However, the relative contribution of these skin DC subsets to the induction of immune responses after Ag targeting has not been addressed in vivo. We show in this study that murine epidermal LCs and dermal DCs transport intradermally injected mAbs against the lectin receptor DEC-205/CD205 in vivo. Skin DCs targeted in situ with mAbs migrated through lymphatic vessels in steady state and inflammation. In the skin-draining lymph nodes, targeting mAbs were found in resident CD8α(+) DCs and in migrating skin DCs. More than 70% of targeted DCs expressed Langerin, including dermal Langerin(+) DCs and LCs. Numbers of targeted skin DCs in the nodes increased 2-3-fold when skin was topically inflamed by the TLR7 agonist imiquimod. Complete removal of the site where OVA-coupled anti-DEC-205 had been injected decreased endogenous cytotoxic responses against OVA peptide-loaded target cells by 40-50%. Surprisingly, selective ablation of all Langerin(+) skin DCs in Langerin-DTR knock-in mice did not affect such responses independently of the adjuvant chosen. Thus, in cutaneous immunization strategies where Ag is targeted to DCs, Langerin(+) skin DCs play a major role in transport of anti-DEC-205 mAb, although Langerin(neg) dermal DCs and CD8α(+) DCs are sufficient to subsequent CD8(+) T cell responses.


Evidence of accelerated evolution and ectodermal-specific expression of presumptive BDS toxin cDNAs from Anemonia viridis.
Anemonia viridis is a widespread and extensively studied Mediterranean species of sea anemone from which a large number of polypeptide toxins, such as blood depressing substances (BDS) peptides, have been isolated. The first members of this class, BDS-1 and BDS-2, are polypeptides belonging to the β-defensin fold family and were initially described for their antihypertensive and antiviral activities. BDS-1 and BDS-2 are 43 amino acid peptides characterised by three disulfide bonds that act as neurotoxins affecting Kv3.1, Kv3.2 and Kv3.4 channel gating kinetics. In addition, BDS-1 inactivates the Nav1.7 and Nav1.3 channels. The development of a large dataset of A. viridis expressed sequence tags (ESTs) and the identification of 13 putative BDS-like cDNA sequences has attracted interest, especially as scientific and diagnostic tools. A comparison of BDS cDNA sequences showed that the untranslated regions are more conserved than the protein-coding regions. Moreover, the KA/KS ratios calculated for all pairwise comparisons showed values greater than 1, suggesting mechanisms of accelerated evolution. The structures of the BDS homologs were predicted by molecular modelling. All toxins possess similar 3D structures that consist of a triple-stranded antiparallel β-sheet and an additional small antiparallel β-sheet located downstream of the cleavage/maturation site; however, the orientation of the triple-stranded β-sheet appears to differ among the toxins. To characterise the spatial expression profile of the putative BDS cDNA sequences, tissue-specific cDNA libraries, enriched for BDS transcripts, were constructed. In addition, the proper amplification of ectodermal or endodermal markers ensured the tissue specificity of each library. Sequencing randomly selected clones from each library revealed ectodermal-specific expression of ten BDS transcripts, while transcripts of BDS-8, BDS-13, BDS-14 and BDS-15 failed to be retrieved, likely due to under-representation in our cDNA libraries. The calculation of the relative abundance of BDS transcripts in the cDNA libraries revealed that BDS-1, BDS-3, BDS-4, BDS-5 and BDS-6 are the most represented transcripts.


Isolation and purification of viable Ureaplasma urealyticum cells free from medium components.
A procedure was devised to produce Ureaplasma urealyticum preparations free of adsorbed components of the growth medium, which contains high concentrations of serum. The ureaplasmas were cultivated in a medium containing PPLO-serum fraction as a replacement for horse serum. High titres of ureaplasmas (greater than 10(7) c.f.u. ml-1) were obtained. Harvested cells were then purified by Urografin density gradient centrifugation. By use of 3H-labelled ureaplasma cells and 125I-labelled medium components, a distinct band of viable cells devoid of serum constituents was demonstrated. The absence of medium components was verified by immunoblotting cells from this band with antiserum to medium components. Medium components that had been present before the purification procedure were undetectable in the purified cell fraction obtained. The viability of the purified ureaplasma cells represented an 85% recovery rate and their antigenicity, examined with anti-serotype specific antiserum, remained intact. This easy and reproducible procedure can be used to prepare purified ureaplasmas for investigation of ureaplasmal antigens and their expression and/or role in disease.


Impact of Sample Collection Order on the Diagnostic Performance of Metagenomic Deep Sequencing for Infectious Keratitis.
The purpose of this article was to evaluate the impact of sample collection order on the diagnostic yield of metagenomic deep sequencing (MDS) for determining the causative pathogen in infectious keratitis. We performed a cross-sectional diagnostic test evaluation among subjects with infectious keratitis at Aravind Eye Hospital in Madurai, India. All subjects underwent corneal scrapings of the affected eye to obtain potassium hydroxide smear, Gram stain, bacterial culture, and fungal culture, in this order. The order of MDS specimen collection relative to smear and culture samples was randomized and served as the primary predictor. Outcomes included the normalized copy number of pathogenic RNA detected by MDS, the proportion of MDS samples that were diagnostic, and the agreement of MDS results with cultures. MDS samples from 46 subjects with corneal ulcers were evaluated. MDS was positive in 33 subjects (76%) and had 74% overall agreement with culture results. There was no association between order of MDS sample collection and normalized copy number of genetic material detected (P = 0.62) or the likelihood of MDS positivity (P = 0.46). However, the likelihood of agreement between MDS and cultures decreased when MDS corneal swabs were collected after other diagnostic corneal scrapings (P = 0.05). The overall yield of MDS for detecting the cause of infectious keratitis was not affected by sample collection order. However, diagnostic agreement between MDS and cultures decreased when MDS samples were collected after other specimens. Additional investigation is warranted to determine whether this represents increased sensitivity of MDS compared with cultures or higher susceptibility to contaminants.


Diversity and regional inequalities in the Spanish 'system of health care services'.
The consolidation of a universal health system coupled with a process of regional devolution characterise the institutional reforms of the National Health System (NHS) in Spain in the last two decades. However, scarce empirical evidence has been reported on the effects of changes in health inputs, outputs and outcomes, both at the country and at the regional level. This paper examines health care reform in Spain along with empirical evidence on regional diversity, efficiency and inequality of these changes in the Spanish NHS. Results suggest that besides significant heterogeneity, once region-specific needs are taken into account, there is evidence of efficiency improvements whilst inequalities in inputs and outcomes, although more 'visible', do not appear to have increased in the last decade. Therefore, the devolution process in the Spanish Health System offers an interesting case for the experimentation of health reforms related to regional diversity but compatible with the nature of a public NHS, with no sizeable regional inequalities.


Comment on "An early Miocene extinction in pelagic sharks".
Sibert and Rubin (Reports, 4 June 2021, p. 1105) claim to have identified a previously unidentified, major extinction event of open-ocean sharks in the early Miocene. We argue that their interpretations are based on an experimental design that does not account for a considerable rise in the sedimentation rate coinciding with the proposed event, nor for intraspecific variation in denticle morphology.


[Bilateral loss of the horizontal vestibulo-ocular reflex in Wernicke encephalopathy - a specific and early clinical sign].
Wernicke encephalopathy is a condition in which thiamine deficiency causes cognitive dysfunction, ataxia and eye motor abnormalities. The condition is potentially reversible if treated with high doses of intravenous thiamine (vitamin B1). The video head impulse test, in which the vestibulo-ocular reflex is tested, can reveal an early specific sign of selective dysfunction in the medial vestibular nuclei by demonstrating selective dysfunction of the horisontal vestibulo-ocular reflex. We present two cases with clinical and neuroradiological findings consistent with Wernicke encephalopathy where such oculomotor findings were detected.


Modelling the spatiotemporal complexity of interactions between pathogenic bacteria and a phage with a temperature-dependent life cycle switch.
We apply mathematical modelling to explore bacteria-phage interaction mediated by condition-dependent lysogeny, where the type of the phage infection cycle (lytic or lysogenic) is determined by the ambient temperature. In a natural environment, daily and seasonal variations of the temperature cause a frequent switch between the two infection scenarios, making the bacteria-phage interaction with condition-dependent lysogeny highly complex. As a case study, we explore the natural control of the pathogenic bacteria Burkholderia pseudomallei by its dominant phage. B. pseudomallei is the causative agent of melioidosis, which is among the most fatal diseases in Southeast Asia and across the world. We assess the spatial aspect of B. pseudomallei-phage interactions in soil, which has been so far overlooked in the literature, using the reaction-diffusion PDE-based framework with external forcing through daily and seasonal parameter variation. Through extensive computer simulations for realistic biological parameters, we obtain results suggesting that phages may regulate B. pseudomallei numbers across seasons in endemic areas, and that the abundance of highly pathogenic phage-free bacteria shows a clear annual cycle. The model predicts particularly dangerous soil layers characterised by high pathogen densities. Our findings can potentially help refine melioidosis prevention and monitoring practices.


Ebola Entry Inhibitors Discovered from Maesa perlarius.
Ebola virus disease (EVD), a disease caused by infection with Ebola virus (EBOV), is characterized by hemorrhagic fever and a high case fatality rate. With limited options for the treatment of EVD, anti-Ebola viral therapeutics need to be urgently developed. In this study, over 500 extracts of medicinal plants collected in the Lingnan region were tested against infection with Ebola-virus-pseudotyped particles (EBOVpp), leading to the discovery of Maesa perlarius as an anti-EBOV plant lead. The methanol extract (MPBE) of the stems of this plant showed an inhibitory effect against EBOVpp, with an IC50 value of 0.52 µg/mL, which was confirmed by testing the extract against infectious EBOV in a biosafety level 4 laboratory. The bioassay-guided fractionation of MPBE resulted in three proanthocyanidins (procyanidin B2 (1), procyanidin C1 (2), and epicatechin-(4β→8)-epicatechin-(4β→8)-epicatechin-(4β→8)-epicatechin (3)), along with two flavan-3-ols ((+)-catechin (4) and (-)-epicatechin (5)). The IC50 values of the compounds against pseudovirion-bearing EBOV-GP ranged from 0.83 to 36.0 µM, with 1 as the most potent inhibitor. The anti-EBOV activities of five synthetic derivatives together with six commercially available analogues, including EGCG ((-)-epigallocatechin-3-O-gallate (8)), were further investigated. Molecular docking analysis and binding affinity measurement suggested the EBOV glycoprotein could be a potential molecular target for 1 and its related compounds.


[The features of uric acid metabolism in women in climacterium].
The purpose of the study was to establish the prevalence of uric acid (UA) metabolism disturbances as a possible risk factor of urate nephropathy in women of various age groups. The authors carried out a retrospective analysis of 757 case histories and performed uricemia and uricosuria screening in women aged 18 to 69 years. The study revealed significant age-dependent elevation of uricemia, with hyperuricemia (HUE) at the age of 60 to 69 years old. The study also found age-dependent increase of HUE rate with its significant elevation in postmenopausal women (older than 50 years), compared with HUE rate in reproductive age women (18 to 49 years old) -37.9% vs. 11.4%. For the first time the researchers established UA excretion level and found a significant tendency of uricosuria to decrease with age (p < 0.016 between the groups of 40 to 49 and 50 to 59 years old). The highest rate of hyperuricosuria was revealed in women of 40 to 49 years old (22.2%), while in women of 18 to 29 and 60 to 69 years old it was 10.1% to 12.1%. For the first time the authors established the rate of hypouricosuria in women, which culminated in women older than 50 (22% of cases). The increase of HUE rate, which is most prominent in postmenopausal women, is associated with the decrease in UA excretion, accompanied by the decrease of hyperuricosuria rate and increase of hypouricosuria rate. Taking into account high prevalence of UA metabolism disturbances in women older than 50, menopause may be considered a risk factor of HUE and urate nephropathy.


[GRADE guidelines 20: Assessing the certainty of evidence in the importance of outcomes or values and preferences - inconsistency, imprecision, and other domains].
To provide Grading of Recommendations, Assessment, Development, and Evaluation (GRADE) guidance for assessing inconsistency, imprecision, and other domains for the certainty of evidence about the relative importance of outcomes. We applied the GRADE domains to rate the certainty of evidence in the importance of outcomes to several systematic reviews, iteratively reviewed draft guidance, and consulted GRADE members and other stakeholders for feedback. We describe the rationale for considering the remaining GRADE domains when rating the certainty in a body of evidence for the relative importance of outcomes. As meta-analyses are not common in this context, inconsistency and imprecision assessments are challenging. Furthermore, confusion exists about inconsistency, imprecision, and true variability in the relative importance of outcomes. To clarify this issue, we suggest that the true variability is neither equivalent to inconsistency nor imprecision. Specifically, inconsistency arises from population, intervention, comparison and outcome and methodological elements that should be explored and, if possible, explained. The width of the confidence interval and sample size inform judgments about imprecision. We also provide suggestions on how to detect publication bias and discuss the domains to rate up the certainty. We provide guidance and examples for rating inconsistency, imprecision, and other domains for a body of evidence describing the relative importance of outcomes.


Systemic therapy for rosacea: focus on oral antibiotic therapy and safety.
Although potentially significant adverse reactions and drug interactions have been reported in association with erythromycin, oral tetracyclines, and trimethoprim-sulfamethoxazole, overall these agents are associated with excellent safety profiles, especially considering their widespread use over many years. It must be considered that when these antibiotics are used for the treatment of rosacea and also for acne vulgaris, their use is on a long-term basis rather than their typical short-course regimens for most infectious diseases. As a result, dermatologists prescribing these agents may feel assured that most patients will not encounter any significant problems, but they do need to be aware of potential adverse reactions to allow for early recognition and discontinuation of the offending drug when needed. Early recognition also allows for favorable management of adverse reactions. In addition, potentially significant drug interactions may be recognized by obtaining a thorough medical history and avoiding combinations of drugs that may interact unfavorably. Fortunately, there are several choices that allow us to individually select a treatment regimen that is optimal for the individual patient, allowing for effective control of rosacea.


Advances in Integrating Traditional and Omic Biomarkers When Analyzing the Effects of the Mediterranean Diet Intervention in Cardiovascular Prevention.
Intervention with Mediterranean diet (MedDiet) has provided a high level of evidence in primary prevention of cardiovascular events. Besides enhancing protection from classical risk factors, an improvement has also been described in a number of non-classical ones. Benefits have been reported on biomarkers of oxidation, inflammation, cellular adhesion, adipokine production, and pro-thrombotic state. Although the benefits of the MedDiet have been attributed to its richness in antioxidants, the mechanisms by which it exercises its beneficial effects are not well known. It is thought that the integration of omics including genomics, transcriptomics, epigenomics, and metabolomics, into studies analyzing nutrition and cardiovascular diseases will provide new clues regarding these mechanisms. However, omics integration is still in its infancy. Currently, some single-omics analyses have provided valuable data, mostly in the field of genomics. Thus, several gene-diet interactions in determining both intermediate (plasma lipids, etc.) and final cardiovascular phenotypes (stroke, myocardial infarction, etc.) have been reported. However, few studies have analyzed changes in gene expression and, moreover very few have focused on epigenomic or metabolomic biomarkers related to the MedDiet. Nevertheless, these preliminary results can help to better understand the inter-individual differences in cardiovascular risk and dietary response for further applications in personalized nutrition.


Reducing Trap-Assisted Recombination in Small Organic Molecule-Based Photovoltaics by the Addition of a Conjugated Block Copolymer.
The performance of organic photovoltaics (OPVs) based on the small-molecule organic semiconductor p-DTS(FBTTh2 )2 is greatly improved by the addition of a conjugated block copolymer composed of difluoroquinoxaline and thienopyrrolodione blocks (D130). The power conversion efficiency (PCE) of the p-DTS(FBTTh2 )2 -based OPV is improved from 5.08% to 6.75% by the addition of 5 wt% D130 to the photoactive layer, which is composed of p-DTS(FBTTh2 )2 and a fullerene derivative. Current-voltage and grazing incidence wide-angle X-ray scattering analyses revealed that the addition of D130 significantly reduces the trap density of the device and changes the packing orientation of p-DTS(FBTTh2 )2 from mostly edge-on to partially face-on. These changes greatly improve the charge carrier mobility of the OPV, indicating that D130 is highly compatible with p-DTS(FBTTh2 )2 . Furthermore, the addition of D130 improve the photostability of the OPV by reducing the burn-in loss under a light soaking intensity of 1 sun. The D130-based OPV maintained 34% of its initial PCE after a light soaking test for 858 h. In contrast, the PCE of the OPV without D130 reduced to 14% of its initial efficiency in the same time period.


Development of kojic acid loaded collagen-chitosan nanoparticle as skin lightener product: in vitro and in vivo assessment.
In the present study, an ionic gelation and ultrasonic approach was performed to produce kojic acid (KA) loaded chitosan(CS)/collagen(CN) nanoparticle(NP) (CSCN-NP) which aimed to increase the dermal delivery and anti-pigmentation effect. To optimize the CSCN-NP the effect of the amount of CN was investigated. The results showed that increasing CN from 0 to 500 mg increased the mean particle size and entrapment efficiency of KA-CSCN-NP from 266.07 ± 9.30 nm to 404.23 ± 9.44 nm and 17.37 ± 2.06% to 82.34 ± 2.16%, respectively. Differential scanning calorimetry confirmed the amorphous form of KA in CSCN-NP, while scanning electron microscopy revealed that the nanoparticles were spherical. There was no chemical interaction between KA and the other components base on attenuated total reflectance-Fourier transform infrared spectroscopy. The skin permeability test showed that KA-CSCN-NP gel delivered more KA to the dermal layers (29.16 ± 1.67% or 537.26 ± 537.26 μg/cm2) and receiver compartment (15.04 ± 1.47% or 277.15 ± 27.22 μg/cm2) compared to KA plain gel. In vitro cytotoxicity assay demonstrated that the improved KA-CSCN-NP was non-toxic. Dermal irritating test on Wistar rats showed that the KA gel was non-irritating. Furthermore, KA-CSCN-NP was found to inhibit melanin formation to a greater extent than free KA and significantly inhibited L-dopa auto-oxidation (94.80 ± 2.41%) compared to pure kojic acid solution (75.28 ± 3.22%). The observations of this study revealed that the produced KA-CSCN-NP might be used as a potential nano-vehicle for KA dermal administration, thereby opening up innovative options for the management of hyper-melanogenesis problems.


An Unrecognized Function of Cholesterol: Regulating the Mechanism Controlling Membrane Phospholipid Asymmetry.
An asymmetric distribution of phospholipids in the membrane bilayer is inseparable from physiological functions, including shape preservation and survival of erythrocytes, and by implication other cells. Aminophospholipids, notably phosphatidylserine (PS), are confined to the inner leaflet of the erythrocyte membrane lipid bilayer by the ATP-dependent flippase enzyme, ATP11C, counteracting the activity of an ATP-independent scramblase. Phospholipid scramblase 1 (PLSCR1), a single-transmembrane protein, was previously reported to possess scrambling activity in erythrocytes. However, its function was cast in doubt by the retention of scramblase activity in erythrocytes of knockout mice lacking this protein. We show that in the human erythrocyte PLSCR1 is the predominant scramblase and by reconstitution into liposomes that its activity resides in the transmembrane domain. At or below physiological intracellular calcium concentrations, total suppression of flippase activity nevertheless leaves the membrane asymmetry undisturbed. When liposomes or erythrocytes are depleted of cholesterol (a reversible process in the case of erythrocytes), PS quickly appears at the outer surface, implying that cholesterol acts in the cell as a powerful scramblase inhibitor. Thus, our results bring to light a previously unsuspected function of cholesterol in regulating phospholipid scrambling.


Hopping in the Crowd to Unveil Network Topology.
We introduce a nonlinear operator to model diffusion on a complex undirected network under crowded conditions. We show that the asymptotic distribution of diffusing agents is a nonlinear function of the nodes' degree and saturates to a constant value for sufficiently large connectivities, at variance with standard diffusion in the absence of excluded-volume effects. Building on this observation, we define and solve an inverse problem, aimed at reconstructing the a priori unknown connectivity distribution. The method gathers all the necessary information by repeating a limited number of independent measurements of the asymptotic density at a single node, which can be chosen randomly. The technique is successfully tested against both synthetic and real data and is also shown to estimate with great accuracy the total number of nodes.


Ocular immune responses. II. Priming of A/J mice in the vitreous induces either enhancement of or suppression of subsequent hapten-specific DTH responses.
We previously showed that the "immunologic privilege" of the anterior chamber results not from afferent blockade, but rather from induction of hapten-specific suppressor T cells that appear after anterior chamber priming with antigen. These suppressor T cells induced by anterior chamber priming differ from those induced by i.v. priming in their ability to block the efferent as well as the afferent limb of the immune response and in their lack of idiotypic surface receptors detected by rabbit anti-idiotypic antibody. We now report that intravitreal priming with haptenated syngeneic spleen cells does not result in generation of suppressor cells, but rather, can in some instances result in an enhanced systemic immunoreactivity to the priming hapten. If soluble antigenic preparations are used, however, intravitreal priming results in the generation of the suppressor T cells, which suppress subsequent DTH and CTL responses to the immunizing hapten. The suppressor cell generation after intravitreal priming is a cyclophosphamide-sensitive process. These results demonstrate that soluble products are processed differently in ocular compartments compared with cell surface coupled ligands, and further demonstrate that the generation of hapten-specific suppressor T cells is dependent, at least in part, on the form and on the specific compartment of the eye that is inoculated.


Metabolism of tetrahydrobiopterin: its relevance in monoaminergic neurons and neurological disorders.
(6R)-L-erythro-5,6,7,8-tetrahydrobiopterin (BH4) is an essential cofactor for aromatic amino acid hydroxylases, such as phenylalanine hydroxylase (PAH), tyrosine hydroxylase (TH), tryptophan hydroxylase, and nitric oxide synthase, which catalyze physiologically important reactions in mammals. The biosynthesis and metabolism of BH4 is usually studied mostly in the liver and only slightly in the brain, as the BH4 level in the liver is relatively high because BH4 is required for the reaction of PAH. We found that GTP (guanosine triphosphate) cyclohydrolase I, an enzyme for the biosynthesis of BH4, is a causative gene for DOPA (3,4-dihydroxyphenylalanine)-responsive dystonia (also called Segawa's disease), and that partial deficiency of BH4 leads to the dysfunction of the nigrostriatal dopaminergic neurons without hyperphenylalaninemia. We analyzed BH4-deficient mice that were produced by disruption of a BH4-synthesizing gene by a gene-knockout technique. We found that the protein amount of TH was highly dependent on the amount of BH4, especially in nerve terminals. Our research suggests that BH4 metabolism in the brain should be different from that in the liver, and that altered metabolism of BH4 should lead to neuropsychiatric disorders including Parkinson's disease.


Brain Functional Magnetic Resonance Imaging Highlights Altered Connections and Functional Networks in Patients With Hypertension.
Hypertension is one of the main risk factors for vascular dementia and Alzheimer disease. To predict the onset of these diseases, it is necessary to develop tools to detect the early effects of vascular risk factors on the brain. Resting-state functional magnetic resonance imaging can investigate how the brain modulates its resting activity and analyze how hypertension impacts cerebral function. Here, we used resting-state functional magnetic resonance imaging to explore brain functional-hemodynamic coupling across different regions and their connectivity in patients with hypertension, as compared to subjects with normotension. In addition, we leveraged multimodal imaging to identify the signature of hypertension injury on the brain. Our study included 37 subjects (18 normotensives and 19 hypertensives), characterized by microstructural integrity by diffusion tensor imaging and cognitive profile, who were subjected to resting-state functional magnetic resonance imaging analysis. We mapped brain functional connectivity networks and evaluated the connectivity differences among regions, identifying the altered connections in patients with hypertension compared with subjects with normotension in the (1) dorsal attention network and sensorimotor network; (2) dorsal attention network and visual network; (3) dorsal attention network and frontoparietal network. Then we tested how diffusion tensor imaging fractional anisotropy of superior longitudinal fasciculus correlates with the connections between dorsal attention network and default mode network and Montreal Cognitive Assessment scores with a widespread network of functional connections. Finally, based on our correlation analysis, we applied a feature selection to highlight those most relevant to describing brain injury in patients with hypertension. Our multimodal imaging data showed that hypertensive brains present a network of functional connectivity alterations that correlate with cognitive dysfunction and microstructural integrity. Registration- URL: https://www.clinicaltrials.gov; Unique identifier: NCT02310217.


Exploring the genetic and adaptive diversity of a pan-Mediterranean crop wild relative: narrow-leafed lupin.
This first pan-Mediterranean analysis of genetic diversity in wild narrow-leafed lupin revealed strong East-West genetic differentiation of populations, an historic eastward migration, and signatures of genetic adaptation to climatic variables. Most grain crops suffer from a narrow genetic base, which limits their potential for adapting to new challenges such as increased stresses associated with climate change. Plant breeders are returning to the wild ancestors of crops and their close relatives to broaden the genetic base of their crops. Understanding the genetic adaptation of these wild relatives will help plant breeders most effectively use available wild diversity. Here, we took narrow-leafed lupin (Lupinus angustifolius L.) as a model to understand adaptation in a wild crop ancestor. A set of 142 wild accessions of narrow-leafed lupin from across the Mediterranean basin were subjected to genotyping-by-sequencing using Diversity Arrays Technology. Phylogenetic, linkage disequilibrium and demographic analyses were employed to explore the history of narrow-leafed lupin within the Mediterranean region. We found strong genetic differentiation between accessions from the western and eastern Mediterranean, evidence of an historic West to East migration, and that eastern Mediterranean narrow-leafed lupin experienced a severe and recent genetic bottleneck. We showed that these two populations differ for flowering time as a result of local adaptation, with the West flowering late while the East flowers early. A genome-wide association study identified single nucleotide polymorphism markers associated with climatic adaptation. Resolving the origin of wild narrow-leafed lupin and how its migration has induced adaptation to specific regions of the Mediterranean serves as a useful resource not only for developing narrow-leafed lupin cultivars with greater resilience to a changing climate, but also as a model which can be applied to other legumes.


Adipogenic genes expression in relation to hepatic steatosis in the liver of two duck species.
Some studies have shown that expression of peroxisome proliferator-activated receptor gamma (PPARG), a key regulator of adipogenesis, and of some adipocyte-specific genes or adipokines are expressed in hepatic steatosis, leading to the concept of 'adipogenic hepatic steatosis' or 'hepatic adiposis.' Most of these studies were conducted in genetic obese mouse models or after manipulation of gene expression. The relevance of this concept to other species and more physiological models was here addressed in ducks which are able to develop hepatic steatosis after overfeeding. The expression of PPARG and other adipocyte-specific genes was thus analyzed in the liver of ducks fed ad libitum or overfed and compared with those observed in adipose tissues. Pekin (Anas platyrhynchos) and Muscovy ducks (Cairina moschata) were analyzed, as metabolic responses to overfeeding differ according to these two species, Muscovy ducks having a greater ability to synthesize and store lipids in the liver than Pekin ducks. Our results indicate that adipocyte-specific genes are expressed in the liver of ducks, PPARG and fatty acid-binding protein 4 being upregulated and adiponectin and leptin receptor downregulated by overfeeding. However, these expression levels are much lower than those observed in adipose tissue suggesting that fatty liver cells are not transformed to adipocytes, although some hepato-specific functions are decreased in fatty liver when compared with normal liver.


Serum transferrin as a marker of bone growth in boys: correlation with serum alkaline phosphatase activity, plasma insulin-like growth factor 1 and rate of growth in height.
Transferrin (TF) has a growth promoting activity in cell culture. The aim of this work was to study possible relationships between serum TF, alkaline phosphatase activity (ALP), plasma insulin-like growth factor 1 (IGF-1) levels and rate of height increase in boys. 149 boys aged 13-15 yrs were studied. TF levels were measured using turbidimetric method. The serum levels of ALP could be used as a biochemical marker for bone formation. Significant correlation was found between serum TF levels and ALP levels (r = 0.31, P less than 0.0005). The TF levels are higher in iron-deficiency anemia. The hemoglobin (Hb) and serum ferritin were measured in all boys. Thirty-one of 149 boys had no iron-deficiency anemia (Hb 14.0 g/dl and serum ferritin 23 ng/ml). The rate of growth in height was estimated over a 5 month period. In these boys, the rate of growth in height was significantly correlated with serum TF levels (r = 0.37; P less than 0.05). A significant correlation was found between serum TF levels and plasma IGF-1 levels (r = 0.45; P less than 0.05). These data indicate that serum TF levels may be a marker of skeletal growth in normal boys.


Bioavailability of griseofulvin from tablets in beagle dogs and correlation with dissolution rate and bioavailability in humans.
The bioavailability of four griseofulvin tablets in beagle dogs, including an ultramicrosize tablet used previously in a human bioavailability study, was investigated on the basis of the plasma 6-demethyl-griseofulvin concentration. The relations with the in vivo findings in humans and the in vitro dissolution rates also were examined. Contrary to the lower bioavailability of the ultramicrosize formulation in humans, it provided the best bioavailability in beagles. The microsize griseofulvin formulations showed similar in vivo results to those in humans. Poor correlation of in vivo parameters between humans and beagles was attributed to the discrepancy of the availability of the ultramicrosize formulation between the two species. The dissolution rates determined by the pretreatment method using plastic beads were correlated more with the in vivo findings than those determined by the other methods. Beagles were a useful animal model for bioavailability studies of certain griseofulvin formulations but not ultramicrosize ones.


X-ray diffraction and in vivo studies reveal the quinary structure of Trypanosoma cruzi nucleoside diphosphate kinase 1: a novel helical oligomer structure.
Trypanosoma cruzi is a flagellated protozoan parasite that causes Chagas disease, which represents a serious health problem in the Americas. Nucleoside diphosphate kinases (NDPKs) are key enzymes that are implicated in cellular energy management. TcNDPK1 is the canonical isoform in the T. cruzi parasite. TcNDPK1 has a cytosolic, perinuclear and nuclear distribution. It is also found in non-membrane-bound filaments adjacent to the nucleus. In the present work, X-ray diffraction and in vivo studies of TcNDPK1 are described. The structure reveals a novel, multi-hexameric, left-handed helical oligomer structure. The results of directed mutagenesis studies led to the conclusion that the microscopic TcNDPK1 granules observed in vivo in T. cruzi parasites are made up by the association of TcNDPK1 oligomers. In the absence of experimental data, analysis of the interactions in the X-ray structure of the TcNDPK1 oligomer suggests the probable assembly and disassembly steps: dimerization, assembly of the hexamer as a trimer of dimers, hexamer association to generate the left-handed helical oligomer structure and finally oligomer association in a parallel manner to form the microscopic TcNDPK1 filaments that are observed in vivo in T. cruzi parasites. Oligomer disassembly takes place on the binding of substrate in the active site of TcNDPK1, leading to dissociation of the hexamers. This study constitutes the first report of such a protein arrangement, which has never previously been seen for any protein or NDPK. Further studies are needed to determine its physiological role. However, it may suggest a paradigm for protein storage reflecting the complex mechanism of action of TcNDPK1.


Overview of toll-like receptors in the CNS.
Mammalian Toll-like receptors (TLRs) were first identified in 1997 based on their homology with Drosophila Toll, which mediates innate immunity in the fly. Over the past eight years, the number of manuscripts describing TLR expression and function in the central nervous system (CNS) has been increasing steadily and expanding beyond their traditional roles in infectious diseases to neurodegenerative disorders and injury. Interest in the field serves as the impetus for this volume in the Current Topics in Microbiology and Immunology series entitled Toll-Like Receptors: Roles in Infection and Neuropathology. The first five chapters highlight more traditional roles for TLRs in infectious diseases of the CNS. The second half of the volume discusses recently emerging roles for TLRs in noninfectious neurodegenerative diseases and the challenges faced by these models in identifying endogenous ligands. Several conceptual theories are introduced in various chapters that deal with the dual nature of TLR engagement and whether these signals favor neuroprotective versus neurodegenerative outcomes.


The association of statin use and statin type and cognitive performance: analysis of the reasons for geographic and racial differences in stroke (REGARDS) study.
Statin use and type have been variably associated with impaired or improved cognitive performance. To assess the association of statin use and type (lipophilic vs hydrophilic) and cognitive impairment. Cross-sectional analysis of 24 595 participants (7191 statin users and 17 404 nonusers) age > or = 45 years, from a population-based national cohort study (Reasons for Geographic And Racial Differences in Stroke) enrolled between January 2003 and October 2008, with oversampling from the southeastern Stroke Belt and African Americans. Statin use and type were documented in participants' homes by a trained health professional. Cognitive performance was assessed with a prior validated instrument of global cognitive status (Six-Item Screener). Cognitive impairment was defined as a score of < 4. Overall, an association of cognitive impairment and statin use was observed (8.6% of users vs 7.7% of nonusers had cognitive impairment, P = 0.014); but, after adjusting for variables known to be associated with cognition (age, gender, race, income, education level, and cardiovascular disease), the association was attenuated (odds ratio [OR]: 0.98, confidence interval [CI]: 0.87-1.10). No association was observed between statin type (lipophilic vs hydrophilic) and cognition (OR: 1.03, CI: 0.86-1.24), and there were no regional differences in cognitive impairment in statin users (8% in the Stroke Belt and 7.9% in other regions, P = 0.63). Statin use and type were marginally associated with cognitive impairment. After adjusting for known variables that affect cognition, no association was observed. No regional differences were observed. This large study found no evidence to support an association between statins and cognitive performance.


DNMT3L modulates significant and distinct flanking sequence preference for DNA methylation by DNMT3A and DNMT3B in vivo.
The DNTM3A and DNMT3B de novo DNA methyltransferases (DNMTs) are responsible for setting genomic DNA methylation patterns, a key layer of epigenetic information. Here, using an in vivo episomal methylation assay and extensive bisulfite methylation sequencing, we show that human DNMT3A and DNMT3B possess significant and distinct flanking sequence preferences for target CpG sites. Selection for high or low efficiency sites is mediated by the base composition at the -2 and +2 positions flanking the CpG site for DNMT3A, and at the -1 and +1 positions for DNMT3B. This intrinsic preference reproducibly leads to the formation of specific de novo methylation patterns characterized by up to 34-fold variations in the efficiency of DNA methylation at individual sites. Furthermore, analysis of the distribution of signature methylation hotspot and coldspot motifs suggests that DNMT flanking sequence preference has contributed to shaping the composition of CpG islands in the human genome. Our results also show that the DNMT3L stimulatory factor modulates the formation of de novo methylation patterns in two ways. First, DNMT3L selectively focuses the DNA methylation machinery on properly chromatinized DNA templates. Second, DNMT3L attenuates the impact of the intrinsic DNMT flanking sequence preference by providing a much greater boost to the methylation of poorly methylated sites, thus promoting the formation of broader and more uniform methylation patterns. This study offers insights into the manner by which DNA methylation patterns are deposited and reveals a new level of interplay between members of the de novo DNMT family.


Combination of converging collimators for high-sensitivity brain SPECT.
The objective of this study, which is related to human brain SPECT, was to increase the sensitivity of a triple-camera SPECT system and reduce statistical noise in reconstructed images using a combination of converging collimators. The reason for combining collimators is to ensure both high sensitivity and sufficient sampling without trading off spatial resolution. A high-sensitivity half-cone-beam (HCB) collimator, designed specifically for brain imaging, was combined with other collimators and compared with conventional parallel-beam and fanbeam circular orbit acquisitions. For comparison, previously studied HCB collimation with a circle-and-helix data acquisition trajectory was also included in this study. Simulations of the Hoffman 3-dimensional brain phantom were performed to calculate the efficiencies of collimators and their combinations and to quantitatively evaluate reconstruction bias, statistical noise, and signal-to-noise ratios in the reconstructed images. Experimental brain phantom data were also acquired and compared for different acquisition types. Finally, a patient brain scan was obtained with a combination of HCB and fanbeam collimators and compared with a triple-fanbeam circular orbit acquisition. A combination of 2 HCB collimators and 1 fanbeam collimator, compared with a triple-fanbeam collimator, can increase the photon detection efficiency by 27% and by more than a factor of 2, compared with triple-parallel-hole collimation, with equal spatial resolution measured on the axis of rotation. Quantitative analysis of reconstruction bias and visual analysis of the images showed no signs of sampling artifacts. Reconstructed images in the simulations, experimental brain phantom, and patient brain scans showed improved quality with this collimator combination due to increased sensitivity and reduced noise. Lesion visibility was also improved, as confirmed by signal-to-noise ratios. Alternatively, triple-HCB circle-and-helix acquisition has also shown competitive results, with a slight disadvantage in axial sampling and implementation procedure. Combined HCB and fanbeam collimation is a promising approach for high-sensitivity brain SPECT.


Defining the role of cholecystokinin in the lipid-induced human brain activation matrix.
In human beings, as in most vertebrates, the release of the intestinal peptide cholecystokinin (CCK) by ingested food plays a major role both in digestion and the regulation of further food intake, but the changes in brain function and their underlying activation mechanisms remain unknown. Our aim was to explore, using a novel physiologic magnetic resonance imaging approach, the temporospatial brain activation matrix, in response to ingestion of a lipid meal and, by use of a CCK-1 receptor antagonist, to define the role of CCK in this activation. We studied, in 19 healthy subjects, the brain activation responses to ingested lipid (dodecanoic acid) or saline (control) with magnetic resonance imaging. Gallbladder volume, plasma CCK levels, and subjective hunger and fullness scores were also recorded. The experiment was then repeated, with and without prior administration of the CCK-1 receptor antagonist dexloxiglumide (600 mg orally) with a controlled, randomized order, latin-square design. Ingested lipid activated bilaterally a matrix of brain areas, particularly the brain stem, pons, hypothalamus, and also cerebellum and motor cortical areas. These activations were abolished by dexloxiglumide, indicating a CCK-mediated pathway, independent of any nutrient-associated awareness cues. The identification of these activations now defines the lipid-activated brain matrix and provides a means by which the gut-derived homeostatic mechanisms of food regulation can be distinguished from secondary sensory and hedonic cues, thereby providing a new approach to exploring aberrant human gastrointestinal responses and eating behavior.


Opportunities for use of radiation therapy in penile cancer based on patterns of care in the United States from 2007 to 2013.
Radiation therapy (RT) is an effective modality for the treatment of squamous cell carcinomas of the penis. The National Comprehensive Cancer Network recommends consideration of primary radiation for penile preservation, in surgically unresectable tumors, and as adjuvant therapy for positive margins, bulky groin nodes or pelvic nodes. We performed a population-based analysis to evaluate the usage of RT in penile cancer from 2007 to 2013. We used the Surveillance, Epidemiology and End Results (SEER) database to identify men diagnosed with squamous cell carcinoma of the penis from 2007 to 2013. Patients were grouped as early stage (T1-T2N0), locally advanced (T3-T4N0), node-positive (T1xN1-3) and metastatic. We used linear regression model to test for factors associated with adjuvant radiation in node-positive patients. We identified 2200 men diagnosed with penile cancer between 2007 and 2013. Of these, 66.4% had early stage, 10.7% had locally advanced, 15.5% had node-positive, 3.2% had metastatic cancer. Among patient with early stage cancer, RT was used in 14 patients (1.0%) and postoperative radiation in an additional 45 patients (3.1%). Among 340 patients with node-positive cancer, 62.1% received surgery alone, 5.6% radiation alone, 21.8% surgery with adjuvant radiation, and 10.6% neither surgery nor radiation. Of patients who had surgery, 26.0% had adjuvant radiation. On univariate analysis, higher nodal stage (N2-3 versus N1) was associated with adjuvant radiation (p = 0.02), while there was a trend for higher T-stage (T3/T4 versus T1/T2) (p = 0.08) and history of prior malignancy (p = 0.06). On multivariate analysis, only higher nodal stage (N2-3 versus N1) was associated with use of adjuvant radiation [hazard ratio (HR) 1.94, p = 0.03]. A small percentage of patient who are eligible for primary or adjuvant RT in the United States receive this treatment. Further work should be done to assess barriers to use of radiation in patients with penile cancer.


Analysis of demographic factors, clinical and investigation findings of patients with pancreatic adenocarcinoma who presented to a tertiary referral center in Sri Lanka.
Pancreatic cancer has a high mortality and morbidity and its incidence has increased over the years. Our objective was to analyze the demographic and clinical data among pancreatic adenocarcinoma patients who presented to the University Surgical Unit, CSTH from 2018 to 2021. Of the 103 patients included, majority were males above the age of 60. The commonest presentation was epigastric pain (54%). 46% were resectable at the time of diagnosis but 50% of the study population were lost to follow up due to a multitude of reasons.


Evolution of molecular error rates and the consequences for evolvability.
Making genes into gene products is subject to predictable errors, each with a phenotypic effect that depends on a normally cryptic sequence. Many cryptic sequences have strongly deleterious effects, for example when they cause protein misfolding. Strongly deleterious effects can be avoided globally by avoiding making errors (e.g., via proofreading machinery) or locally by ensuring that each error has a relatively benign effect. The local solution requires powerful selection acting on every cryptic site and so evolves only in large populations. Small populations with less effective selection evolve global solutions. Here we show that for a large range of realistic intermediate population sizes, the evolutionary dynamics are bistable and either solution may result. The local solution facilitates the genetic assimilation of cryptic genetic variation and therefore substantially increases evolvability.


Glucocorticoid receptor and serum- and glucocorticoid-induced kinase-1 in esophageal adenocarcinoma and adjacent Barrett's esophagus.
Barrett's esophagus (BE) is a consequence of gastroesophageal reflux disease and is predisposed to esophageal adenocarcinoma (EAC). EAC is an exemplar model of inflammation-associated cancer. Glucocorticoids suppress inflammation through glucocorticoid receptor (GR) and serum- and glucocorticoid-induced kinase-1 (Sgk1) expressions. Therefore, we immunolocalized GR and Sgk1 in EAC and the adjacent BE tissues and studied their association with clinical disease course in 87 patients with EAC who underwent surgical resection (N = 58) or endoscopic submucosal dissection (N = 29). Low GR and Sgk1 expressions in adjacent BE tissues were associated with adverse clinical outcomes (P = 0.0008 and 0.034, respectively). Patients with low Sgk1 expression in EAC cells exhibited worse overall survival (P = 0.0018). In multivariate Cox regression analysis, low GR expression in the adjacent nonmalignant BE tissues was significantly associated with worse overall survival (P = 0.023). The present study indicated that evaluation of GR and Sgk1 expressions in both the EAC cells and adjacent nonmalignant BE tissues could help to predict clinical outcomes following endoscopic and surgical treatments. In particular, the GR status in BE tissues adjacent to EAC was an independent prognostic factor.


Mitochondrial energy metabolism is required for lifespan extension by the spastic paraplegia-associated protein spartin.
Hereditary spastic paraplegias, a group of neurodegenerative disorders, can be caused by loss-of-function mutations in the protein spartin. However, the physiological role of spartin remains largely elusive. Here we show that heterologous expression of human or Drosophila spartin extends chronological lifespan of yeast, reducing age-associated ROS production, apoptosis, and necrosis. We demonstrate that spartin localizes to the proximity of mitochondria and physically interacts with proteins related to mitochondrial and respiratory metabolism. Interestingly, Nde1, the mitochondrial external NADH dehydrogenase, and Pda1, the core enzyme of the pyruvate dehydrogenase complex, are required for spartin-mediated cytoprotection. Furthermore, spartin interacts with the glycolysis enhancer phospo-fructo-kinase-2,6 (Pfk26) and is sufficient to complement for PFK26-deficiency at least in early aging. We conclude that mitochondria-related energy metabolism is crucial for spartin's vital function during aging and uncover a network of specific interactors required for this function.


Pallidal Deep Brain Stimulation Improves Higher Control of the Oculomotor System in Parkinson's Disease.
The frontal cortex and basal ganglia form a set of parallel but mostly segregated circuits called cortico-basal ganglia loops. The oculomotor loop controls eye movements and can direct relatively simple movements, such as reflexive prosaccades, without external help but needs input from "higher" loops for more complex behaviors. The antisaccade task requires the dorsolateral prefrontal cortex, which is part of the prefrontal loop. Information flows from prefrontal to oculomotor circuits in the striatum, and directional errors in this task can be considered a measure of failure of prefrontal control over the oculomotor loop. The antisaccadic error rate (AER) is increased in Parkinson's disease (PD). Deep brain stimulation (DBS) of the subthalamic nucleus (STN) has no effect on the AER, but a previous case suggested that DBS of the globus pallidus interna (GPi) might. Our aim was to compare the effects of STN DBS and GPi DBS on the AER. We tested eye movements in 14 human DBS patients and 10 controls. GPi DBS substantially reduced the AER, restoring lost higher control over oculomotor function. Interloop information flow involves striatal neurons that receive cortical input and project to pallidum. They are normally silent when quiescent, but in PD they fire randomly, creating noise that may account for the degradation in interloop control. The reduced AER with GPi DBS could be explained by retrograde stimulation of striatopallidal axons with consequent activation of inhibitory collaterals and reduction in background striatal firing rates. This study may help explain aspects of PD pathophysiology and the mechanism of action of GPi DBS. Significance statement: Parkinson's disease causes symptoms including stiffness, slowness of movement, and tremor. Electrical stimulation of specific areas deep in the brain can effectively treat these symptoms, but exactly how is not fully understood. Part of the cause of such symptoms may be impairments in the way information flows from one circuit within the brain to another, as a result of overactivity of certain nerve cells. By demonstrating that stimulation of an area called the globus pallidus interna partially reverses deficits in voluntary control of eye movements, this study shows that stimulation can improve information flow between circuits, probably by calming down the overactive cells.


A rapid feedback signal is not always necessary for termination of a drinking bout.
When a pig is deprived of drinking water, a deficit of body water develops that is corrected when the pig drinks to satiation. If food is available during the deprivation, the stimulus to drinking is plasma hyperosmolality. Because of the delay in correction of plasma hyperosmolality as ingested water is slowly absorbed, it has been thought that a rapid inhibitory signal from the digestive tract is necessary to prevent overdrinking. This concept was tested by measuring changes in plasma osmolality before and during drinking after such deprivation and also after infusion of hypertonic saline. As drinking began, there was a rapid fall of plasma osmolality to levels insufficient to drive drinking by the time drinking ended. This fall of plasma hyperosmolality to subthreshold levels while the pig is drinking seems to make a rapid inhibitory control signal from the digestive tract unnecessary to terminate the drinking bout under these conditions.


In search of non-random X inactivation: studies of fetal membranes heterozygous for glucose-6-phosphate dehydrogenase.
Extraembryonic membranes and fetal tissues were obtained from 55 specimens of 5--11 weeks conceptual age. The glucose-6-phosphate dehydrogenase (G6PD) electrophoretic phenotype was determined and correlated with that of maternal blood. Fifteen specimens were heterozygous for G6PD A, and for nine of these the maternal allele could be determined. In none of these specimens did the isozyme pattern of the membraneous chorion or chorionic villi differ significantly from that of fetal tissue. We have obtained no evidence of non-random inactivation in extraembryonic membranes of human fetal specimens at this stage of development.