NTP Toxicology and Carcinogenesis Studies of HC Red No. 3 [2,((Amino-2-nitrophenyl)amino)ethanol] (CAS No. 2871-01-4) in F344/N Rats and B6C3F1 Mice (Gavage Studies).
oxicology and carcinogenesis studies of HC Red No. 3 (97% pure), a semipermanent hair dye, were conducted by administering the chemical in corn oil by gavage for 105 weeks to groups of 50 male and 50 female F344/N rats and for 104 weeks to groups of 50 male and 50 female B6C3F1 mice. The dosage regimen used for rats was 0, 250, or 500 mg/kg per day and for mice, 0, 125. or 250 mg/kg per day. Doses were administered 5 days per week. In prior 13-week studies, these doses produced no signs of toxicity when administered 5 days per week. In the 2-year studies, the administration of HC Red No. 3 did not affect body weight gains of male or female rats or mice. Body weight gains by all groups of female mice were reduced because of a reproductive tract infection. Survival of male and female rats and mice was not reduced by administration of HC Red No. 3. The survival of female mice, including vehicle controls, was reduced relative to historical survival rates due to a reproductive tract infection. The infection, accompanied by weight loss, high mortality, and suppurative inflammation of multiple organs, was found in 36/50 vehicle control, 32/50 low dose, and 29/50 high dose female mice. Klebsiella pneumoniae was isolated from infected tissues. Pigmentation of various tissues in both rats and mice was a common observation in both the 13-week and the 2-year studies. The pigment was not identified but was presumed to be a derivative of HC Red No. 3. Very minimal nephropathy was found in dosed female rats, but its relationship to HC Red No. 3 is equivocal. Mild nephrosis was found in dosed female mice, but this effect may have been secondary to the infection of the genital tract. There was an increase in the incidence of mammary gland fibroadenomas or cystadenomas in low dose female rats. The incidence of this lesion in high dose female rats was not increased (vehicle control, 14/50, 28%; low dose, 25/50, 50%; high dose, 11/50, 22%). Largely because of the lack of a dose response, the increased incidence in the low dose females was not considered to be due to HC Red No. 3. No increased incidences of neoplasms were seen in male rats. Transitional cell papillomas of the urinary bladder were detected in one high dose male rat, two low dose female rats, and one high dose female rat; none was observed in the vehicle controls. These uncommon neoplasms were found in animals that survived to the termination of the study and were not accompanied by other proliferative lesions. The incidence of hepatocellular adenomas or carcinomas (combined) was increased in high dose male mice, whereas the incidence of these neoplasms in low dose male mice was significantly lower than that in the vehicle controls (25/50; 15/50; 35/50). Hepatocellular carcinomas in three vehicle control, one low dose, and five high dose male mice metastasized to the lung. The incidences of liver neoplasms in dosed female mice were not significantly different from those in the vehicle control group. HC Red No. 3 was mutagenic in Salmonella typhimurium strains TA97, TA98, and TA100, but not in TA1535, in the presence or absence of Aroclor 1254-induced male Sprague-Dawley rat or male Syrian hamster liver S9 when tested by the preincubational protocol. An audit of the experimental data was conducted for these 2-year toxicology and carcinogenesis studies on HC Red No. 3. No data discrepancies were found that influenced the final interpretations. Under the conditions of these 2-year gavage studies of HC Red No. 3, there was no evidence of carcinogenicity for male or female F344/N rats given 250 or 500 mg/kg per day. There was equivocal evidence of carcinogenicity for male B6C3F1 mice as indicated by an increased incidence of hepatocellular adenomas or carcinomas (combined) in the 250 mg/kg dose group. Poor survival coupled with lack of significant findings rendered the study in female B6C3F1 mice an inadequate study of carcinogenicity. Both sexes of both species may have been able to tolerate higher doses of HC Red No. 3. Therefore, the sensitivity of these studies for detecting chese studies for detecting carcinogenesis may have been limited. Synonym: 2,((amino-2-nitrophenyl)amino)ethanol


System-level study on synergism and antagonism of active ingredients in traditional Chinese medicine by using molecular imprinting technology.
In this work, synergism and antagonism among active ingredients of traditional Chinese medicine (TCM) were studied at system-level by using molecular imprinting technology. Reduning Injection (RDNI), a TCM injection, was widely used to relieve fever caused by viral infection diseases in China. Molecularly imprinted polymers (MIPs) synthesized by sol-gel method were used to separate caffeic acid (CA) and analogues from RDNI without affecting other compounds. It can realize the preparative scale separation. The inhibitory effects of separated samples of RDNI and sample combinations in prostaglandin E2 biosynthesis in lipopolysaccharide-induced RAW264.7 cells were studied. The combination index was calculated to evaluate the synergism and antagonism. We found that components which had different scaffolds can produce synergistic anti-inflammatory effect inside and outside the RDNI. Components which had similar scaffolds exhibited the antagonistic effect, and the antagonistic effects among components could be reduced to some extent in RDNI system. The results indicated MIPs with the characteristics of specific adsorption ability and large scale preparation can be an effective approach to study the interaction mechanism among active ingredients of complex system such as TCM at system-level. And this work would provide a new idea to study the interactions among active ingredients of TCM.


The RopGEF KARAPPO Is Essential for the Initiation of Vegetative Reproduction in Marchantia polymorpha.
Many plants can reproduce vegetatively, producing clonal progeny from vegetative cells; however, little is known about the molecular mechanisms underlying this process. Liverwort (Marchantia polymorpha), a basal land plant, propagates asexually via gemmae, which are clonal plantlets formed in gemma cups on the dorsal side of the vegetative thallus [1]. The initial stage of gemma development involves elongation and asymmetric divisions of a specific type of epidermal cell, called a gemma initial, which forms on the floor of the gemma cup [2, 3]. To investigate the regulatory mechanism underlying gemma development, we focused on two allelic mutants in which no gemma initial formed; these mutants were named karappo, meaning "empty." We used whole-genome sequencing of both mutants and molecular genetic analysis to identify the causal gene, KARAPPO (KAR), which encodes a ROP guanine nucleotide exchange factor (RopGEF) carrying a plant-specific ROP nucleotide exchanger (PRONE) catalytic domain. In vitro GEF assays showed that the full-length KAR protein and the PRONE domain have significant GEF activity toward MpROP, the only ROP GTPase in M. polymorpha. Moreover, genetic complementation experiments showed a significant role for the N- and C-terminal variable regions in gemma development. Our investigation demonstrates an essential role for KAR/RopGEF in the initiation of plantlet development from a differentiated cell, which may involve cell-polarity formation and subsequent asymmetric cell division via activation of ROP signaling, implying a similar developmental mechanism in vegetative reproduction of various land plants.


Intermediate tunnelling-hopping regime in DNA charge transport.
Charge transport in molecular systems, including DNA, is involved in many basic chemical and biological processes, and its understanding is critical if they are to be used in electronic devices. This important phenomenon is often described as either coherent tunnelling over a short distance or incoherent hopping over a long distance. Here, we show evidence of an intermediate regime where coherent and incoherent processes coexist in double-stranded DNA. We measure charge transport in single DNA molecules bridged to two electrodes as a function of DNA sequence and length. In general, the resistance of DNA increases linearly with length, as expected for incoherent hopping. However, for DNA sequences with stacked guanine-cytosine (GC) base pairs, a periodic oscillation is superimposed on the linear length dependence, indicating partial coherent transport. This result is supported by the finding of strong delocalization of the highest occupied molecular orbitals of GC by theoretical simulation and by modelling based on the Büttiker theory of partial coherent charge transport.


Restorative Mechanisms Regulating Protein Balance in Skeletal Muscle During Recovery From Sepsis.
Muscle deconditioning is commonly observed in patients surviving sepsis. Little is known regarding the molecular mechanisms regulating muscle protein homeostasis during the recovery or convalescence phase. We adapted a sepsis-recovery mouse model that uses cecal ligation and puncture (CLP), followed 24 h later by cecal resection and antibiotic treatment, to identify putative cellular pathways regulating protein synthesis and breakdown in skeletal muscle. Ten days after CLP, body weight and food consumption did not differ between control and sepsis-recovery mice, but gastrocnemius weight was reduced. During sepsis-recovery, muscle protein synthesis was increased 2-fold and associated with enhanced mTOR kinase activity (4E-BP1 and S6K1 phosphorylation). The sepsis-induced increase in 4E-BP1 was associated with enhanced formation of the eIF4E-eIF4G active cap-dependent complex, while the increased S6K1 was associated with increased phosphorylation of downstream targets S6 and eIF4B. Proximal to mTOR, sepsis-recovery increased Akt and TSC2 phosphorylation, did not alter AMPK phosphorylation, and decreased REDD1 protein content. Despite the decreased mRNA content for the E3 ubiquitin ligases atrogin-1 and muscle RING-finger 1, proteasomal activity was increased 50%. In contrast, sepsis-recovery was associated with an apparent decrease in autophagy (e.g., increased ULK-1 phosphorylation, decreased LCB3-II, and increased p62). The mRNA content for IL-1β, IL-18, TNFα, and IL-6 in muscle was elevated in sepsis-recovery. During recovery after sepsis skeletal muscle responds with an increase in Akt-TSC2-mTOR-dependent protein synthesis and decreased autophagy, but full restoration of muscle protein content may be slowed by the continued stimulation of ubiquitin-proteasome activity.


Gangliocytic paraganglioma of the cauda equina with significant calcification: first description in pediatric age.
A 17-year-old boy was referred with a 2-month history of low back pain and bilateral sciatica and difficulty in ambulation. Neurological examination found mild muscle weakness and diminished deep tendon reflexes in his right leg. Lumbar magnetic resonance imaging revealed an intradural-extramedullary tumor at the level of the L4, exiting through the right L4-L5 intervertebral foramen into the right psoas muscle. After total resection of the tumor, histopathological diagnosis revealed a gangliocytic paraganglioma. There are 184 paraganglioma cases reported at the lumbar region to date, and only 4 of them were in the pediatric age group. This report is the fifth paraganglioma case in the lumbar region and the first gangliocytic paraganglioma case in the pediatric age population.


Vitamin B6 in treatment of tardive dyskinesia: a preliminary case series study.
Tardive dyskinesia (TD) remains a significant problem for patients and physicians. Several reports have suggested that vitamin B6 (pyridoxine) can be helpful in the treatment of some neuroleptic-induced movement disorders, including parkinsonism and TD. This report presents the results of a preliminary study of five patients with TD who underwent a four week open-label clinical trial of vitamin B6 (100 mg/d) in addition to their regular medications. The severity of the involuntary movements was assessed using the Abnormal Involuntary Movement Scale (AIMS), Barnes Akathisia Rating Scale (BARS) and the Simpson-Angus Scale (SAS). The patients' clinical status was assessed with the Brief Psychiatric Rating Scale (BPRS). With the addition of vitamin B6 to their treatment, four patients had clinically significant (greater than 30%) improvement on the measures of involuntary movement and, in three cases, there was also clinically significant improvement on the BPRS. None of the patients had side effects attributable to vitamin B6. The results suggest that vitamin B6 may alleviate TD, but it will need to be further tested in controlled double-blind trials.


Middle-Column Gap Balancing and Middle-Column Mismatch in Spinal Reconstructive Surgery.
Middle-column gap balancing (MCGB) is a reference measurement of the path of the posterior longitudinal ligament (PLL), which is reconstructed under tension and balanced by the combined height of the posterior one-third of the vertebral bodies and the posterior one-third of the disks, including any intervening load-sharing spacers. This measurement allows for a comparison of the ligamentous component of the middle column (PLL) with the load-sharing components (posterior one-third vertebral body + disk ). This difference gives rise to a "middle-column mismatch," which provides a linear measurement of the redundancy of the ligaments and neural elements, which relates to the correct cage, spacer, or load-bearing height, which is optimized. For phase 1 measurement testing, 24 consecutive patients underwent reliable flexion, extension, and neutral lateral radiographic studies with a calibrated marker. The anterior, middle, and posterior columns were measured using a custom software program capable of measuring the length of curved lines specifically written for this purpose. For phase 2 measurement testing, 21 consecutive patients undergoing surgery with multilevel deformities for cervical, thoracic, and lumbar procedures had MCGB height pre- and postoperatively measured by 3 blinded observers. The preoperative and postoperative measurements were compared using a linear regression analysis and Pearson product-moment correlation. In phase 1 measurement testing the flexion, extension, and neutral bending radiographs of spinal segments not containing deformities showed that the middle column had the most reliable measurements of spinal axial height both in the actual measurements of change from flexion to extension (mm) and in percentage of change. In phase 2 measurement testing, a Pearson product-moment correlation was run between each individual's pre- and postoperative middle-column measurements. There was a strong positive correlation between preoperative and postoperative measurements, which was statistically significant (r = 0.983, n = 21, P < .01). This consecutive series of 21 deformity patients demonstrated the utility of measuring the preoperative middle-column length in predicting the optimal height of the spacers and intervertebral disks, and posterior vertebral body height, simultaneously restoring sagittal and coronal plane alignment. Key points of this study include the following: (1) Spinal balance requires optimizing spinal height, which is a curved line in order to accommodate cervical lordosis, thoracic kyphosis, and lumbar lordosis. (2) Software programs can allow measurement of the preoperative curved circuitous course of the PLL and vertebral body misalignment; this curved length is predictive of the optimal postoperative middle-column height after spinal osteotomies and intervertebral spacer insertion. (3) All 3 dimensions are important to optimize in deformity correction: sagittal plane, coronal plane, and axial spinal height.


[Odontoblast: a key cell involved in the perception of dentinal pain].
Dentinal sensitivity is a clinical condition daily encountered by practitioners and constitutes the symptoms of dentinal hypersensitivity, a common dental pain affecting on average 30% of the population. However, the management of this pathology is not always effective due to the lack of knowledge particularly concerning the means by which dental nociceptive signals are transduced. The mechanisms underlying dentin sensitivity still remain unclear probably due to the structural and functional complexity of the players including odontoblasts, nerve endings and dentinal fluid running in the dentinal tubules. The unique spatial situation of odontoblasts, ciliated cells in close relationship with nerve terminals, suggests that they could play a pivotal role in the transduction of sensory events occurring within the dentin tissue. Our studies have identified mechano-thermosensitive transient receptor potential ion channels (TRPV1-4, TRPA8, TRPM3, KCa, TREK-1, PC1, PC2) localised on the odontoblastic membrane and at the base of the cilium. They could sense temperature variations or movements of dentinal fluid within tubules. Moreover, several voltage-gated sodium channels confer excitable properties to odontoblasts in response to injection of depolarizing currents. In vivo, these channels co-localize with nerve endings at the apical pole of odontoblasts, and their expression pattern seems to be correlated with the spatial distribution of stretch-activated KCa channels. All these data strengthen the hypothesis that odontoblasts could act as sensor cells able to transmit nociceptive signals. However, how cells sense signals and how the latter are transmitted to axons represent the main issue to be solved.


Arranging ISO 13606 archetypes into a knowledge base.
To enable the efficient reuse of standard based medical data we propose to develop a higher level information model that will complement the archetype model of ISO 13606. This model will make use of the relationships that are specified in UML to connect medical archetypes into a knowledge base within a repository. UML connectors were analyzed for their ability to be applied in the implementation of a higher level model that will establish relationships between archetypes. An information model was developed using XML Schema notation. The model allows linking different archetypes of one repository into a knowledge base. Presently it supports several relationships and will be advanced in future.


Mosaic trisomy 13: understanding origin using SNP array.
Trisomy 13 occurs in 1/10,000-20,000 live births, and mosaicism accounts for 5% of these cases. Phenotype and outcome of mosaic trisomy 13 are variable and poorly understood. Microsatellite analyses of trisomy 13 have indicated the high incidence of maternal meiotic origin and reduced recombination, but no study has focused on mosaic trisomy 13 in live born patients. Single-nucleotide polymorphism (SNP) array, fluorescence in situ hybridisation and bioinformatics analyses were performed in three cases of mosaic trisomy 13. Two cases of complete mosaic trisomy 13 originated from meiosis I non-disjunction followed by trisomic rescue; one had crossovers resulting in segmental uniparental disomy in the disomic line, and one had no crossover. Mosaicism for partial trisomy 13 in the third complex case either arose from meiosis II non-disjunction without crossover or in early mitosis followed by anaphase lags. The extra chromosome 13 was maternal in origin in all three cases. Mosaicism percentage calculated from B allele frequencies ranged from 30 to 50. Genotypes and copy number information provided by SNP array allow determination of parental origin and uniparental disomy status and direct quantification of mosaicism. Such information may lead to a better understanding of mechanisms underlying mosaic aneuploidies and the observed phenotypic variability and better prediction of recurrent risk.


Associations Between Surgical Ablation and Operative Mortality After Mitral Valve Procedures.
Surgical ablation (SA) for atrial fibrillation (AF) concomitant to mitral valve repair/replacement (MVRR) improves longitudinal sinus rhythm. However, the risk of adding SA remains a clinical question. This study examined whether the addition of contemporary SA for AF has an impact on operative outcomes. The study cohort included 88,765 MVRR patients with or without SA, coronary artery bypass grafting (CABG), septal defect, and tricuspid repair in The Society of Thoracic Surgeons Database between 2011 and 2014. Group 1 did not have AF (No-AF) and did not receive SA (No-SA); group 2 had No-AF immediately preoperatively but received SA; group 3 had AF but No-SA; and group 4 had AF with SA. Groups 3 and 4 were stratified into paroxysmal versus nonparoxysmal AF. With the use of logistic regression, with group 1 as reference, risk-adjusted odds ratios (OR) for mortality were compared for SA performance, AF type, and SA technique. Group 3 had higher age, New York Heart Association class, redo operations, and unadjusted mortality than group 4. Relative to group 1, group 3 had an OR for mortality of 1.15 (95% confidence interval: 1.04 to 1.27, p < 0.01). OR increments were similar for paroxysmal and nonparoxysmal AF. In group 4, concomitant SA was independently associated with lower AF-related relative risk (OR 1.08), to a level that was not different from group 1 (p = 0.13). Observed treatment effects were equivalent for paroxysmal and nonparoxysmal AF and across all levels of baseline risk. For patients with AF at the time of mitral operation, the performance of SA seems associated with a lower risk-adjusted operative mortality compared with patients who do not undergo ablation.


Electron Accumulative Molecules.
With the goal to produce molecules with high electron accepting capacity and low reorganization energy upon gaining one or more electrons, a synthesis procedure leading to the formation of a B-N(aromatic) bond in a cluster has been developed. The research was focused on the development of a molecular structure able to accept and release a specific number of electrons without decomposing or change in its structural arrangement. The synthetic procedure consists of a parallel decomposition reaction to generate a reactive electrophile and a synthesis reaction to generate the B-N(aromatic) bond. This procedure has paved the way to produce the metallacarboranylviologen [M(C2B9H11)(C2B9H10)-NC5H4-C5H4N-M'(C2B9H11)(C2B9H10)] (M = M' = Co, Fe and M = Co and M' = Fe) and semi(metallacarboranyl)viologen [3,3'-M(8-(NC5H4-C5H4N-1,2-C2B9H10)(1',2'-C2B9H11)] (M = Co, Fe) electron cumulative molecules. These molecules are able to accept up to five electrons and to donate one in single electron steps at accessible potentials and in a reversible way. By targeted synthesis and corresponding electrochemical tests each electron transfer (ET) step has been assigned to specific fragments of the molecules. The molecules have been carefully characterized, and the electronic communication between both metal centers (when this situation applies) has been definitely observed through the coplanarity of both pyridine fragments. The structural characteristics of these molecules imply a low reorganization energy that is a necessary requirement for low energy ET processes. This makes them electronically comparable to fullerenes, but on their side, they have a wide range of possible solvents. The ET from one molecule to another has been clearly demonstrated as well as their self-organizing capacity. We consider that these molecules, thanks to their easy synthesis, ET, self-organizing capacity, wide range of solubility, and easy processability, can find important application in any area where ET is paramount.


Determinants of toxicity, patterns of failure, and outcome among adult patients with soft tissue sarcomas of the extremity and superficial trunk treated with greater than conventional doses of perioperative high-dose-rate brachytherapy and external beam radiotherapy.
The present study was undertaken to determine factors predictive of toxicity, patterns of failure, and survival in 60 adult patients with soft tissue sarcomas of the extremity and superficial trunk treated with combined perioperative high-dose-rate brachytherapy and external beam radiotherapy. The patients were treated with surgical resection and perioperative high-dose-rate brachytherapy (16 or 24 Gy) for negative and close/microscopically positive resection margins, respectively. External beam radiotherapy (45 Gy) was added postoperatively to reach a 2-Gy equivalent dose of 62.9 and 72.3 Gy, respectively. Adjuvant chemotherapy with ifosfamide and doxorubicin was given to patients with advanced high-grade tumors. Grade 3 toxic events were observed in 18 patients (30%) and Grade 4 events in 6 patients (10%). No Grade 5 events were observed. A location in the lower limb was significant for Grade 3 or greater toxic events on multivariate analysis (p = .013), and the tissue volume encompassed by the 150% isodose line showed a trend toward statistical significance (p = .086). The local control, locoregional control, and distant control rate at 9 years was 77.4%, 69.5%, and 63.8%, respectively. On multivariate analysis, microscopically involved margins correlated with local control (p = .036) and locoregional control (p = .007) and tumor size correlated with distant metastases (p = .004). The 9-year disease-free survival and overall survival rate was 47.0% and 61.5%, respectively. Multivariate analysis showed poorer disease-free survival rates for patients with tumors >6 cm (p = .005) and microscopically involved margins (p = .043), and overall survival rates decreased with increasing tumor size (p = .011). Grade 3 or greater wound complications can probably be decreased using meticulous treatment planning to decrease the tissue volume encompassed by the 150% isodose line, especially in lower limb locations. Microscopically involved margins remain a predictor of local and locoregional failure, despite radiation doses >70 Gy. Patients with tumors ≥6 cm and microscopically involved margins are at high risk of treatment failure and death from the development of distant metastases.


Reminiscence and adaptation to critical life events in older adults with mild to moderate depressive symptoms.
The role of reminiscence as a way of adapting to critical life events and chronic medical conditions was investigated in older adults with mild to moderate depressive symptoms. Reminiscence is the (non)volitional act or process of recollecting memories of one's self in the past. 171 Dutch older adults with a mean age of 64 years (SD = 7.4) participated in this study. All of them had mild to moderate depressive symptoms. Participants completed measures on critical life events, chronic medical conditions, depressive symptoms, symptoms of anxiety and satisfaction with life. The reminiscence functions included were: identity, problem solving, bitterness revival and boredom reduction. Critical life events were positively correlated with identity and problem solving. Bitterness revival and boredom reduction were both positively correlated with depressive and anxiety symptoms, and negatively to satisfaction with life. Problem solving had a negative relation with anxiety symptoms. When all the reminiscence functions were included, problem solving was uniquely associated with symptoms of anxiety, and bitterness revival was uniquely associated with depressive symptoms and satisfaction with life. Interestingly, problem solving mediated the relation of critical life events with anxiety. This study corroborates the theory that reminiscence plays a role in coping with critical life events, and thereby maintaining mental health. Furthermore, it is recommended that therapists focus on techniques which reduce bitterness revival in people with depressive symptoms, and focus on problem-solving reminiscences among people with anxiety symptoms.


LC-MS/MS confirms that COX-1 drives vascular prostacyclin whilst gene expression pattern reveals non-vascular sites of COX-2 expression.
There are two schools of thought regarding the cyclooxygenase (COX) isoform active in the vasculature. Using urinary prostacyclin markers some groups have proposed that vascular COX-2 drives prostacyclin release. In contrast, we and others have found that COX-1, not COX-2, is responsible for vascular prostacyclin production. Our experiments have relied on immunoassays to detect the prostacyclin breakdown product, 6-keto-PGF1α and antibodies to detect COX-2 protein. Whilst these are standard approaches, used by many laboratories, antibody-based techniques are inherently indirect and have been criticized as limiting the conclusions that can be drawn. To address this question, we measured production of prostanoids, including 6-keto-PGF1α, by isolated vessels and in the circulation in vivo using liquid chromatography tandem mass spectrometry and found values essentially identical to those obtained by immunoassay. In addition, we determined expression from the Cox2 gene using a knockin reporter mouse in which luciferase activity reflects Cox2 gene expression. Using this we confirm the aorta to be essentially devoid of Cox2 driven expression. In contrast, thymus, renal medulla, and regions of the brain and gut expressed substantial levels of luciferase activity, which correlated well with COX-2-dependent prostanoid production. These data are consistent with the conclusion that COX-1 drives vascular prostacyclin release and puts the sparse expression of Cox2 in the vasculature in the context of the rest of the body. In doing so, we have identified the thymus, gut, brain and other tissues as target organs for consideration in developing a new understanding of how COX-2 protects the cardiovascular system.


Nonalcoholic fatty liver disease: current concepts, epidemiology and management strategies.
Nonalcoholic fatty liver disease (NAFLD) is now the most prevalent liver disease in the world. It involves a spectrum of conditions from hepatic steatosis to nonalcoholic steatohepatitis and liver fibrosis, and is a major cause of cirrhosis and hepatocellular carcinoma. It is defined by presence of steatosis in 5% of hepatocytes or more in the absence of other causes of fatty liver. The metabolic syndrome is the major known risk factor for NAFLD. Dietary contributors such as high fructose intake and coffee consumption appear to increase and decrease the risk of disease respectively, but these links are unclear. Genetic associations have also been identified. The estimated prevalence of the disease varies according to diagnostic method and population demographics. It appears to be a major issue in Europe with population studies showing up to 50% of the individuals are affected while in the USA one in three adults are estimated to have NAFLD. Laboratory investigations and ultrasound are typically first-line investigations. Fibrosis may be assessed noninvasively through transient elastography and biomarkers but liver biopsy remains the gold standard to quantify hepatic damage. Associated comorbidities include cardiovascular disease and chronic kidney disease. Weight loss, dietary changes and exercise are recommended in management. Medications should be considered to manage underlying risk factors including insulin resistance. Surgical options include bariatric procedures and liver transplantation. The combination of rising prevalence and significant potential complications warrant further research into NAFLD, particularly in areas with research gaps including Eastern Europe.


Macromolecular crowding directs the motion of small molecules inside cells.
It is now well established that cell interiors are significantly crowded by macromolecules, which impede diffusion and enhance binding rates. However, it is not fully appreciated that levels of crowding are heterogeneous, and can vary substantially between subcellular regions. In this article, starting from a microscopic model, we derive coupled nonlinear partial differential equations for the concentrations of two populations of large and small spherical particles with steric volume exclusion. By performing an expansion in the ratio of the particle sizes, we find that the diffusion of a small particle in the presence of large particles obeys an advection-diffusion equation, with a reduced diffusion coefficient and a velocity directed towards less crowded regions. The interplay between advection and diffusion leads to behaviour that differs significantly from Brownian diffusion. We show that biologically plausible distributions of macromolecules can lead to highly non-Gaussian probability densities for the small particle position, including asymmetrical and multimodal densities. We confirm all our results using hard-sphere Brownian dynamics simulations.


Limited dispersal, budding dispersal, and cooperation: an experimental study.
Numerous theoretical studies have investigated how limited dispersal may provide an explanation for the evolution of cooperation, by leading to interactions between relatives. However, despite considerable theoretical attention, there has been a lack of empirical tests. In this article, we test how patterns of dispersal influence the evolution of cooperation, using iron-scavenging in the bacterium Pseudomonas aeruginosa as our cooperative trait. We found that relatively limited dispersal does not favor cooperation. The reason for this is that although limited dispersal increases the relatedness between interacting individuals, it also leads to increased local competition for resources between relatives. This result supports Taylor's prediction that in the simplest possible scenario, the effects of increased relatedness and local competition exactly cancel out. In contrast, we show that one way for cooperation to be favored is if individuals disperse in groups (budding dispersal), because this maintains high relatedness while reducing local competition between relatives (relatively global competition).


Rotavirus susceptibility of antibiotic-treated mice ascribed to diminished expression of interleukin-22.
The commensal microbiota regulates susceptibility to enteric pathogens by fine-tuning mucosal innate immune responses, but how susceptibility to enteric viruses is shaped by the microbiota remains incompletely understood. Past reports have indicated that commensal bacteria may either promote or repress rotavirus replication in the small intestine of mice. We now report that rotavirus replicated more efficiently in the intestines of germ-free and antibiotic-treated mice compared to animals with an unmodified microbiota. Antibiotic treatment also facilitated rotavirus replication in type I and type III interferon (IFN) receptor-deficient mice, revealing IFN-independent proviral effects. Expression of interleukin-22 (IL-22) was strongly diminished in the intestine of antibiotic-treated mice. Treatment with exogenous IL-22 blocked rotavirus replication in microbiota-depleted wild-type and Stat1-/- mice, demonstrating that the antiviral effect of IL-22 in animals with altered microbiome is not dependent on IFN signaling. In antibiotic-treated animals, IL-22-induced a specific set of genes including Fut2, encoding fucosyl-transferase 2 that participates in the biosynthesis of fucosylated glycans which can mediate rotavirus binding. Interestingly, IL-22 also blocked rotavirus replication in antibiotic-treated Fut2-/- mice. Furthermore, IL-22 inhibited rotavirus replication in antibiotic-treated mice lacking key molecules of the necroptosis or pyroptosis pathways of programmed cell death. Taken together, our results demonstrate that IL-22 determines rotavirus susceptibility of antibiotic-treated mice, yet the IL-22-induced effector molecules conferring rotavirus resistance remain elusive.


Investigation of the relationship between prostate cancer and MSMB and NCOA4 genetic variants and protein expression.
Two genome-wide association studies (GWAS) identified the β-microseminoprotein (MSMB) promoter SNP, rs10993994:C>T, as significantly associated with prostate cancer (PC) risk. Follow-up studies demonstrate that the variant allele directly affects expression of the MSMB-encoded protein, PSP94, and also suggest that it affects mRNA expression levels of an adjacent gene, NCOA4, which is involved in androgen receptor transactivation. In a population-based study of 1,323 cases and 1,268 age-matched controls, we found the NCOA4 SNP, rs7350420:T>C, was associated with a 15% reduction in PC risk, but the association was not significant after adjustment for the rs10993994:C>T genotype. Tumor tissue microarrays of 519 radical prostatectomy patients were used to measure PSP94 and NCOA4 protein expression. Taken together, these data confirm that the rs10993994:C>T variant allele is associated with decreased PSP94 expression, and the association is stronger in tumor compared to normal prostate tissue. No association was observed between rs10993994:C>T and NCOA4 expression, and only moderate associations were seen between two NCOA4 SNPs, rs10761618:T>C and rs7085433:G>A, and NCOA4 protein expression. These data indicate that the increase in PC risk associated with rs10993994:C>T is likely mediated by the variant's effect on PSP94 expression; however, this effect does not extend to NCOA4 in the data presented here.


HER3 and downstream pathways are involved in colonization of brain metastases from breast cancer.
Metastases to the brain from breast cancer have a high mortality, and basal-like breast cancers have a propensity for brain metastases. However, the mechanisms that allow cells to colonize the brain are unclear. We used morphology, immunohistochemistry, gene expression and somatic mutation profiling to analyze 39 matched pairs of primary breast cancers and brain metastases, 22 unmatched brain metastases of breast cancer, 11 non-breast brain metastases and 6 autopsy cases of patients with breast cancer metastases to multiple sites, including the brain. Most brain metastases were triple negative and basal-like. The brain metastases over-expressed one or more members of the HER family and in particular HER3 was significantly over-expressed relative to matched primary tumors. Brain metastases from breast and other primary sites, and metastases to multiple organs in the autopsied cases, also contained somatic mutations in EGFR, HRAS, KRAS, NRAS or PIK3CA. This paralleled the frequent activation of AKT and MAPK pathways. In particular, activation of the MAPK pathway was increased in the brain metastases compared to the primary tumors. Deregulated HER family receptors, particularly HER3, and their downstream pathways are implicated in colonization of brain metastasis. The need for HER family receptors to dimerize for activation suggests that tumors may be susceptible to combinations of anti-HER family inhibitors, and may even be effective in the absence of HER2 amplification (that is, in triple negative/basal cancers). However, the presence of activating mutations in PIK3CA, HRAS, KRAS and NRAS suggests the necessity for also specifically targeting downstream molecules.


Identification and Characterization of Pale Potato Cyst Nematode (Globodera pallida) in Teplá, the Czech Republic.
Potato cyst nematode poses a significant threat to potato producers in the Czech Republic. Both species of potato cyst nematodes (Globodera rostochiensis and G. pallida) are listed as quarantine pests in the Czech Republic and also by the European Union, European and Mediterranean Plant Protection Organization, and North American Plant Protection Organization. To date, G. rostochiensis was responsible for all damage to potatoes caused by cyst nematodes in the Czech Republic, while G. pallida was recorded only once in the Czech Republic (2) 8 years ago. It is important to note that this occurrence of G. pallida was not located in the free-cultivation area. In July 2011, soil samples from a potato field located in the area of Teplá (Karlovy Vary Region) were collected, and the cysts extracted were identified as G. pallida according to microscopic observation of cyst fenestra and morphology of juveniles (1). Cyst morphometrics (means from 10 cysts) included: fenestra diameter 21.2 μm, distance fenestra to anus 56.8 μm, Granek's ratio 2.7, number of cuticular ridges between fenestra and anus 14; while second stage juvenile morphometrics (means of 13 specimens) were: L 466.7 μm, stylet 24.2 μm, tail 53.2 μm, body width at anus 13.2, h 28.6, c 8.8, c' 4.0. Terminus of juvenile tails was rounded, and stylet knobs possessed distinct forward projections. Total DNA was extracted from single cysts using the TriPure reagent (Roche), and the DNA samples were used to amplify cistron rDNA with the following primers: 18S, 5'-TTGATTAGGTCCCTGCCCTTT-3', and 21S, 5'-TTTCACTCGCCGTTACTAAGG-3'. The amplified region contains the 3' end of the 18S gene, ITS1, 5.8S, ITS2, and the 5' end of the 28S gene. Pfu DNA polymerase (Fermentas) was used for accurate amplification. A PCR product of approximately 1.0 kb was amplified from three individual cysts. The PCR amplicons were cloned into pJET1.2 using the CloneJET PCR Cloning Kit (Fermentas) and sequenced in both directions. The sequences of representative isolates were deposited in GenBank (Accession Nos. JQ692592, JQ692593, and JQ692594). The resultant 1.0 kb sequences showed 99% nucleotide identity to sequences of G. pallida from Canada (GenBank Accession Nos. GQ294522.1, GQ355975.1, and GQ294523.1), thus confirming the results of the morphological analyses. To our knowledge, this is the first detection of G. pallida in the free-cultivation area of Karlovy Vary Region, and only the second in the Czech Republic since the first report in 2003 (2). References: (1) M. W. Brzeski. Page 237 in: Nematodes of Tylenchina in Poland and Temperate Europe. Muzeum I Instytut Zoologii Polska Akademia Nauk, Warszawa 1998. (2) M. Zouhar et al. Plant Disease, 87:98, 2003.


Hippocampal gene expression analysis highlights Ly6a/Sca-1 as candidate gene for previously mapped novelty induced behaviors in mice.
In this study, we show that the covariance between behavior and gene expression in the brain can help further unravel the determinants of neurobehavioral traits. Previously, a QTL for novelty induced motor activity levels was identified on murine chromosome 15 using consomic strains. With the goal of narrowing down the linked region and possibly identifying the gene underlying the quantitative trait, gene expression data from this F(2)-population was collected and used for expression QTL analysis. While genetic variation in these mice was limited to chromosome 15, eQTL analysis of gene expression showed strong cis-effects as well as trans-effects elsewhere in the genome. Using weighted gene co-expression network analysis, we were able to identify modules of co-expressed genes related to novelty induced motor activity levels. In eQTL analyses, the expression of Ly6a (a.k.a. Sca-1) was found to be cis-regulated by chromosome 15. Ly6a also surfaced in a group of genes resulting from the network analysis that was correlated with behavior. Behavioral analysis of Ly6a knock-out mice revealed reduced novelty induced motor activity levels when compared to wild type controls, confirming functional importance of Ly6a in this behavior, possibly through regulating other genes in a pathway. This study shows that gene expression profiling can be used to narrow down a previously identified behavioral QTL in mice, providing support for Ly6a as a candidate gene for functional involvement in novelty responsiveness.


Specific gene repression by CRISPRi system transferred through bacterial conjugation.
In microbial communities, bacterial populations are commonly controlled using indiscriminate, broad range antibiotics. There are few ways to target specific strains effectively without disrupting the entire microbiome and local environment. Here, we use conjugation, a natural DNA horizontal transfer process among bacterial species, to deliver an engineered CRISPR interference (CRISPRi) system for targeting specific genes in recipient Escherichia coli cells. We show that delivery of the CRISPRi system is successful and can specifically repress a reporter gene in recipient cells, thereby establishing a new tool for gene regulation across bacterial cells and potentially for bacterial population control.


The origin of surfactant amphiphilicity and self-assembly in protic ionic liquids.
The nature of amphiphilic self-assembly in alkylammonium protic ionic liquids (PILs) is examined by systematically varying the ionic structure and composition, H-bonding capacity, and nanostructure of both the PIL and micelle-forming cationic surfactant, and contrasted with self-assembly in water. Using small-angle neutron scattering, micelle structure and concentrations are determined for primary - quaternary dodecylammonium salts in nitrate and thiocyanate PILs. While the solvophobic driving force depends only on the average polarity of the PIL, surprisingly strong, specific interactions of the head group and counterion with the PIL H-bond network are found. This suggests the importance of developing designer amphiphiles for assembling soft matter structures in PILs.


Acquisition of Streptococcus pneumoniae and nonspecific morbidity in infants and their families: a cohort study.
Most children are believed to acquire Streptococcus pneumoniae asymptomatically, with only a few developing overt S. pneumoniae disease. This study investigates the relationship between acquisition of S. pneumoniae and mild nonspecific infection leading to general practitioner (GP) consultation. A prospective birth cohort study of 213 infants assessed at home 9 times during 24 weeks by nasopharyngeal swab and parental interview was conducted. All positive S. pneumoniae swabs (including acquisition and carriage) were significantly associated with GP consultations for infection by the study infant compared with infants with negative swabs [odds ratio (OR), 1.6; 95% confidence interval (CI) 1.1-2.2; P = 0.005]. There was a stronger association with S. pneumoniae acquisition alone (OR 2.1; 95% CI 1.3-3.4; P = 0.001) than with carriage only (OR 1.4; 95% CI 0.9-2.0; P = 0.1). Multivariate analysis confirmed that S. pneumoniae acquisition by the study subject was independently associated with GP consultations: adjusted hazard ratio, 1.8 (95% CI 1.1-2.9); P = 0.02. A similar and independent association was found between S. pneumoniae acquisition by the study subject, and GP consultations for infection by the family (adjusted hazard ratio, 1.8; 95% CI 1.1-2.8; P = 0.01). Acquisition of S. pneumoniae by the study infant was significantly associated with GP consultations for infection by the infant or family.


Peptides with antimicrobial activity from four different families isolated from the skins of the North American frogs Rana luteiventris, Rana berlandieri and Rana pipiens.
The skins of frogs of the genus Rana synthesize a complex array of antimicrobial peptides that may be grouped into eight families on the basis of structural similarity. A total of 24 peptides with differential growth-inhibitory activity towards the Gram-positive bacterium Staphylococcus aureus, the Gram-negative bacterium Escherichia coli and the yeast Candida albicans were isolated from extracts of the skins of three closely related North American frogs, Rana luteiventris (spotted frog), Rana berlandieri (Rio Grande leopard frog) and Rana pipiens (Northern leopard frog). Structural characterization of the antimicrobial peptides demonstrated that they belonged to four of the known families: the brevinin-1 family, first identified in skin of the Asian frog Rana porosa brevipoda; the esculentin-2 family, first identified in the European frog Rana esculenta; the ranatuerin-2 family, first identified in the North American bullfrog Rana catesbeiana; and the temporin family, first identified in the European frog Rana temporaria. Peptides belonging to the brevinin-2, ranalexin, esculentin-1 and ranatuerin-1 families were not identified in the extracts. Despite the close phylogenetic relationship between the various species of Ranid frogs, the distribution and amino-acid sequences of the antimicrobial peptides produced by each species are highly variable and species-specific, suggesting that they may be valuable in taxonomic classification and molecular phylogenetic analysis.


Residual errors in laser interferometry from air turbulence and nonlinearity.
Laser displacement interferometry is used extensively in precision equipment for semiconductor manufacture. In these applications it is often necessary to introduce a high velocity airflow to the measurement environment to minimize the density of airborne particulate contaminants. The performance of the heterodyne interferometer is degraded by the resulting fluctuations in the index of refraction along the beam path. The magnitude, correlation length, and probability distribution of the optical path length (OPL) fluctuations are measured for several airflow conditions. The data are interpreted in terms of the path length errors for some common interferometric configurations. The OPL fluctuations are generally less significant than the systematic sources of measurement error. A more fundamental limit on the accuracy of the heterodyne Michelson interferometer is the periodic nonlinearity caused by leakage of the frequency components in the beamsplitter. The effect is discussed in detail. A direct observation of the nonlinearity is reported. The magnitude of the effect is about lambda/64 for the beam splitters used in this experiment. A simple technique which indicates the presence and magnitude of the nonlinearity is described.


Preclinical efficacy of azacitidine and venetoclax for infant KMT2A-rearranged acute lymphoblastic leukemia reveals a new therapeutic strategy.
Infants with KMT2A-rearranged B-cell acute lymphoblastic leukemia (ALL) have a dismal prognosis. Survival outcomes have remained static in recent decades despite treatment intensification and novel therapies are urgently required. KMT2A-rearranged infant ALL cells are characterized by an abundance of promoter hypermethylation and exhibit high BCL-2 expression, highlighting potential for therapeutic targeting. Here, we show that hypomethylating agents exhibit in vitro additivity when combined with most conventional chemotherapeutic agents. However, in a subset of samples an antagonistic effect was seen between several agents. This was most evident when hypomethylating agents were combined with methotrexate, with upregulation of ATP-binding cassette transporters identified as a potential mechanism. Single agent treatment with azacitidine and decitabine significantly prolonged in vivo survival in KMT2A-rearranged infant ALL xenografts. Treatment of KMT2A-rearranged infant ALL cell lines with azacitidine and decitabine led to differential genome-wide DNA methylation, changes in gene expression and thermal proteome profiling revealed the target protein-binding landscape of these agents. The selective BCL-2 inhibitor, venetoclax, exhibited in vitro additivity in combination with hypomethylating or conventional chemotherapeutic agents. The addition of venetoclax to azacitidine resulted in a significant in vivo survival advantage indicating the therapeutic potential of this combination to improve outcome for infants with KMT2A-rearranged ALL.


Normal preoperative urodynamic testing does not predict voiding dysfunction after Burch colposuspension versus pubovaginal sling.
Urodynamic studies have been proposed as a means of identifying patients at risk for voiding dysfunction after surgery for stress urinary incontinence. We determined if preoperative urodynamic findings predict postoperative voiding dysfunction after pubovaginal sling or Burch colposuspension. Data were analyzed from preoperative, standardized urodynamic studies performed on participants in the Stress Incontinence Treatment Efficacy Trial, in which women with stress urinary incontinence were randomized to undergo pubovaginal sling surgery or Burch colposuspension. Voiding dysfunction was defined as use of any bladder catheter after 6 weeks, or reoperation for takedown of a pubovaginal sling or Burch colposuspension. Urodynamic study parameters examined were post-void residual urine, maximum flow during noninvasive flowmetry, maximum flow during pressure flow study, change in vesical pressure at maximum flow during pressure flow study, change in abdominal pressure at maximum flow during pressure flow study and change in detrusor pressure at maximum flow during pressure flow study. The study excluded women with a preoperative post-void residual urine volume of more than 150 ml or a maximum flow during noninvasive flowmetry of less than 12 ml per second unless advanced pelvic prolapse was also present. Of the 655 women in whom data were analyzed voiding dysfunction developed in 57 including 8 in the Burch colposuspension and 49 in the pubovaginal sling groups. There were 9 patients who could not be categorized and, thus, were excluded from the remainder of the analysis (646). A total of 38 women used a catheter beyond week 6, 3 had a surgical takedown and 16 had both. All 19 women who had surgical takedown were in the pubovaginal sling group. The statistical analysis of urodynamic predictors is based on subsets of the entire cohort, including 579 women with preoperative uroflowmetry, 378 with change in vesical pressure, and 377 with change in abdominal and detrusor pressure values. No preoperative urodynamic study findings were associated with an increased risk of voiding dysfunction in any group. Mean maximum flow during noninvasive flowmetry values were similar among women with voiding dysfunction compared to those without voiding dysfunction in the entire group (23.4 vs 25.7 ml per second, p = 0.16), in the Burch colposuspension group (25.8 vs 25.7 ml per second, p = 0.98) and in the pubovaginal sling group (23.1 vs 25.7 ml per second, p = 0.17). Voiding pressures and degree of abdominal straining were not associated with postoperative voiding dysfunction. In this carefully selected group preoperative urodynamic studies did not predict postoperative voiding dysfunction or the risk of surgical revision in the pubovaginal sling group. Our findings may be limited by the stringent exclusion criteria and studying a group believed to be at greater risk for voiding dysfunction could alter these findings. Additional analysis using subjective measures to define voiding dysfunction is warranted to further determine the ability of urodynamic studies to stratify the risk of postoperative voiding dysfunction, which appears to be limited in the current study.


A novel mechanism of autophagic cell death in dystrophic muscle regulated by P2RX7 receptor large-pore formation and HSP90.
P2RX7 is an ATP-gated ion channel, which can also exhibit an open state with a considerably wider permeation. However, the functional significance of the movement of molecules through the large pore (LP) and the intracellular signaling events involved are not known. Here, analyzing the consequences of P2RX7 activation in primary myoblasts and myotubes from the Dmd(mdx) mouse model of Duchenne muscular dystrophy, we found ATP-induced P2RX7-dependent autophagic flux, leading to CASP3-CASP7-independent cell death. P2RX7-evoked autophagy was triggered by LP formation but not Ca(2+) influx or MAPK1-MAPK3 phosphorylation, 2 canonical P2RX7-evoked signals. Phosphoproteomics, protein expression inference and signaling pathway prediction analysis of P2RX7 signaling mediators pointed to HSPA2 and HSP90 proteins. Indeed, specific HSP90 inhibitors prevented LP formation, LC3-II accumulation, and cell death in myoblasts and myotubes but not in macrophages. Pharmacological blockade or genetic ablation of p2rx7 also proved protective against ATP-induced death of muscle cells, as did inhibition of autophagy with 3-MA. The functional significance of the P2RX7 LP is one of the great unknowns of purinergic signaling. Our data demonstrate a novel outcome--autophagy--and show that molecules entering through the LP can be targeted to phagophores. Moreover, we show that in muscles but not in macrophages, autophagy is needed for the formation of this LP. Given that P2RX7-dependent LP and HSP90 are critically interacting in the ATP-evoked autophagic death of dystrophic muscles, treatments targeting this axis could be of therapeutic benefit in this debilitating and incurable form of muscular dystrophy.


Virus infection of Haptolina ericina and Phaeocystis pouchetii implicates evolutionary conservation of programmed cell death induction in marine haptophyte-virus interactions.
The mechanisms by which phytoplankton cope with stressors in the marine environment are neither fully characterized nor understood. As viruses are the most abundant entities in the global ocean and represent a strong top-down regulator of phytoplankton abundance and diversity, we sought to characterize the cellular response of two marine haptophytes to virus infection in order to gain more knowledge about the nature and diversity of microalgal responses to this chronic biotic stressor. We infected laboratory cultures of the haptophytes Haptolina ericina and Phaeocystis pouchetii with CeV-01B or PpV-01B dsDNA viruses, respectively, and assessed the extent to which host cellular responses resemble programmed cell death (PCD) through the activation of diagnostic molecular and biochemical markers. Pronounced DNA fragmentation and activation of cysteine aspartate-specific proteases (caspases) were only detected in virus-infected cultures of these phytoplankton. Inhibition of host caspase activity by addition of the pan-caspase inhibitor z-VAD-fmk did not impair virus production in either host-virus system, differentiating it from the Emiliania huxleyi-Coccolithovirus model of haptophyte-virus interactions. Nonetheless, our findings point to a general conservation of PCD-like activation during virus infection in ecologically diverse haptophytes, with the subtle heterogeneity of cell death biochemical responses possibly exerting differential regulation on phytoplankton abundance and diversity.


Prevalence of epilepsy in Rwanda: a national cross-sectional survey.
To estimate the prevalence of epilepsy in Rwanda, as part of a survey of musculoskeletal impairment (MSI). Cross-sectional nationally representative survey. Clusters of 105 people were selected with probability proportional to size, using the 2002 national census as the sampling frame. Within each cluster, 80 people were selected through compact segment sampling. Physiotherapists examined all participants using a standard questionnaire to assess the presence of seizures through self-report, as well as aetiology, treatment needs and severity of the seizures. Health-related quality of life was assessed using the EuroQol generic health index (EQ5D). The response rate was 80.8%, with 6757 individuals screened. The estimated prevalence of epilepsy was 0.7% (95% confidence intervals 0.5-0.9%). A quarter (23%) of cases reported that onset of epilepsy was at birth, for 32% onset was between birth and age 5, 23% between ages 5 and 16, and 21% above the age of 16. In 4.3% of people with epilepsy, an accident during a seizure caused permanent MSI, as identified by a case history and physical examination. The majority of people with epilepsy (59.6%) had never received medical treatment for it. Epilepsy was associated with a significantly reduced quality of life. Epilepsy is a significant and neglected health problem in Rwanda at all ages. Investing in treatment for epilepsy would prevent some disability causing accidents from occurring.


Artificial neural networks and risk stratification in emergency departments.
Emergency departments are characterized by the need for quick diagnosis under pressure. To select the most appropriate treatment, a series of rules to support decision-making has been offered by scientific societies. The effectiveness of these rules affects the appropriateness of treatment and the hospitalization of patients. Analyzing a sample of 1844 patients and focusing on the decision to hospitalize a patient after a syncope event to prevent severe short-term outcomes, this work proposes a new algorithm based on neural networks. Artificial neural networks are a non-parametric technique with the well-known ability to generalize behaviors, and they can thus predict severe short-term outcomes with pre-selected levels of sensitivity and specificity. This innovative technique can outperform the traditional models, since it does not require a specific functional form, i.e., the data are not supposed to be distributed following a specific design. Based on our results, the innovative model can predict hospitalization with a sensitivity of 100% and a specificity of 79%, significantly increasing the appropriateness of medical treatment and, as a result, hospital efficiency. According to Garson's Indexes, the most significant variables are exertion, the absence of symptoms, and the patient's gender. On the contrary, cardio-vascular history, hypertension, and age have the lowest impact on the determination of the subject's health status. The main application of this new technology is the adoption of smart solutions (e.g., a mobile app) to customize the stratification of patients admitted to emergency departments (ED)s after a syncope event. Indeed, the adoption of these smart solutions gives the opportunity to customize risk stratification according to the specific clinical case (i.e., the patient's health status) and the physician's decision-making process (i.e., the desired levels of sensitivity and specificity). Moreover, a decision-making process based on these smart solutions might ensure a more effective use of available resources, improving the management of syncope patients and reducing the cost of inappropriate treatment and hospitalization.


Laboratory parameters associated with prolonged hospital length of stay in COVID-19 patients in Johannesburg, South Africa.
Coronavirus disease (COVID-19) has imposed unprecedented stressors on South Africa (SA)'s healthcare system. Superimposed on the country's quadruple burden of disease, pandemic-related care further exposes existing inequities. Some of these inequities are specific to hospital-based inpatient services, such as the geographical maldistribution of hospital beds, lack of oxygen supplies and assisted ventilation, and scarcity of trained healthcare workers. Certain high-risk groups, such as individuals with cardiometabolic comorbidity, are likely to develop severe COVID-19 disease requiring hospitalisation with potential for a prolonged length of stay (LoS). It may be helpful for health authorities to identify those at risk for prolonged LoS to facilitate appropriate health systems planning. To identify hospital admission laboratory parameters associated with a hospital stay >14 days in patients with COVID-19 pneumonia. A retrospective observational study design was used. Laboratory data were obtained from an SA private laboratory for 642 inpatients with suspected or confirmed COVID-19 pneumonia, comprising 7 months of admission laboratory data from six private hospitals in Johannesburg, Gauteng Province. Of 642 hospital admissions for pneumonia, 497 were confirmed to have COVID-19 infection (reverse transcription-polymerase chain reaction test positive). In the COVID-19-positive group, hospital LoS was prolonged in 35.4% of admissions. Univariate analysis demonstrated an association with the following risk factors for prolonged LoS: older age; male sex; high serum creatinine, sodium (Na), chloride, potassium and urea levels and low estimated glomerular filtration rate; raised white blood cell count, lymphopenia, neutrophilia and an elevated neutrophil-to-lymphocyte ratio (NLR); and elevated levels of D-dimers, interleukin-6 (IL-6), and procalcitonin (PCT). The strongest univariate associations (relative risk (RR) ≥2.0) with a hospital stay >14 days were high Na levels, NRL >18, high PCT levels and IL-6 >40 pg/mL. On multivariable analysis, the following factors remained significantly associated with prolonged LoS: older age (RR 1.015 per year of age; 95% confidence interval (CI) 1.005 - 1.024); hypernatraemia (RR 1.80; 95% CI 1.25 - 2.60); hyperkalaemia (RR 1.61; 95% CI 1.18 - 2.20); and neutrophilia (RR 1.47; 95% CI 1.15 - 1.88). COVID-19 pandemic preparedness requires hospital-based inpatient care to be prioritised in resource-limited settings, and availability of beds and prompt admissions are essential to ensure good clinical outcomes. In this study of COVID-19 patients admitted with pneumonia, multivariable analysis showed older age, hypernatraemia, hyperkalaemia and neutrophilia to be associated with LoS >14 days. This may assist with healthcare systems planning.


The enzymic nature of antibody catalysis: development of multistep kinetic processing.
Detailed kinetic investigations of a catalytic antibody that promotes the hydrolyses of an anilide and phenyl ester show that this catalyst uses a multistep kinetic sequence resembling that found in serine proteases to hydrolyze its substrates, although antibody was elicited to a single transition-state analog. Like the serine proteases the antibody catalyzes the hydrolysis reactions through a putative covalent intermediate, but unlike the enzymes it may use hydroxide ion to cleave the intermediates. Nevertheless, the antibody is a potent catalyst with turnover at higher pH values rivaling that of chymotrypsin. This analysis also reveals that turnover by the antibody is ultimately limited by product desorption, suggesting that improvements in catalytic efficiency may be achieved by judicious changes in the structure of the substrate, so that it is not superimposable on that of the eliciting hapten.


Human arthroplasty derived macrophages differentiate into osteoclastic bone resorbing cells.
In aseptic loosening, a heavy macrophage response to biomaterial wear particles is commonly found in arthroplasty tissues. The aim of this study was to discover if these cells contribute to the bone resorption of aseptic loosening by differentiating into osteoclasts. Macrophages were isolated from the pseudocapsule and pseudomembrane of loose cemented and uncemented hip arthroplasties at the time of revision surgery and then co-cultured on glass coverslips and dentine slices with UMR 106 rat osteoblast-like cells, both in the presence and absence of 1,25 dihydroxyvitamin D3 [1,25(OH)2D3]. Macrophages isolated from the synovial membrane of patients with osteoarthritis (OA) undergoing hip replacements were similarly studied as a control group. After 24 hours incubation, most cells isolated from the above periprosthetic tissues strongly expressed macrophage (CD11b, CD14) but not osteoclast markers. However, after 14 days incubation, numerous multinucleated cells showing the phenotypic features of osteoclasts (that is, positive for tartrate resistant acid phosphatase, the vitronectin receptor, and capable of extensive lacunar resorption) formed in co-cultures of arthroplasty derived macrophages and UMR 106 cells, in the presence of 1,25(OH)2D3. The addition of an antibody to macrophage colony stimulating factor (M-CSF) considerably reduced macrophage-osteoclast differentiation and hence the lacunar resorption seen in these co-cultures. In contrast, OA synovial macrophage/UMR 106 co-cultures showed little or no evidence of macrophage-osteoclast differentiation and this was only seen when human M-CSF was added to the co-cultures. This is the first report showing that human macrophages isolated directly from periprosthetic tissues surrounding loosened implants can differentiate into multinucleated cells showing all the functional and cytochemical characteristics of osteoclasts. In contrast with other macrophage populations, exogenous M-CSF is not required for this to occur. In the context of the heavy macrophage response to wear particles in periprosthetic tissues macrophage-osteoclast differentiation may represent an important cellular mechanism whereby osteolysis is effected in aseptic loosening.


Regulating the HO endonuclease in yeast.
The pedigree of mating-type switching in yeast is determined by the transcription pattern of the HO endonuclease gene, which is expressed during late G1 in mother cells but not at all in daughter cells. The late-G1 specificity of HO transcription depends on a heteromeric factor, SBF, which is composed of the Swi4 and Swi6 proteins. Mother-cell specificity involves a second site-specific DNA-binding factor, Swi5, which is synthesized in the G2 and M phases and only enters the nucleus at the end of mitosis. Swi5 enters mother and daughter nuclei in equal amounts and most is then rapidly degraded. It has been suggested that in mothers but not in daughters some Swi5 protein escapes degradation and persists until SBF is activated in late G1. This subset of Swi5 molecules may constitute a mother cell's memory.


PIM kinases as potential therapeutic targets in a subset of peripheral T cell lymphoma cases.
Currently, there is no efficient therapy for patients with peripheral T cell lymphoma (PTCL). The Proviral Integration site of Moloney murine leukemia virus (PIM) kinases are important mediators of cell survival. We aimed to determine the therapeutic value of PIM kinases because they are overexpressed in PTCL patients, T cell lines and primary tumoral T cells. PIM kinases were inhibited genetically (using small interfering and short hairpin RNAs) and pharmacologically (mainly with the pan-PIM inhibitor (PIMi) ETP-39010) in a panel of 8 PTCL cell lines. Effects on cell viability, apoptosis, cell cycle, key proteins and gene expression were evaluated. Individual inhibition of each of the PIM genes did not affect PTCL cell survival, partially because of a compensatory mechanism among the three PIM genes. In contrast, pharmacological inhibition of all PIM kinases strongly induced apoptosis in all PTCL cell lines, without cell cycle arrest, in part through the induction of DNA damage. Therefore, pan-PIMi synergized with Cisplatin. Importantly, pharmacological inhibition of PIM reduced primary tumoral T cell viability without affecting normal T cells ex vivo. Since anaplastic large cell lymphoma (ALK+ ALCL) cell lines were the most sensitive to the pan-PIMi, we tested the simultaneous inhibition of ALK and PIM kinases and found a strong synergistic effect in ALK+ ALCL cell lines. Our findings suggest that PIM kinase inhibition could be of therapeutic value in a subset of PTCL, especially when combined with ALK inhibitors, and might be clinically beneficial in ALK+ ALCL.


Optofluidic router based on tunable liquid-liquid mirrors.
We present an electrically tunable 1 × 5 optofluidic router for on-chip light routing. The device can redirect light from an optical input channel into five output channels by exploiting total internal reflection (TIR) at a liquid-liquid interface. The liquid-liquid mirrors, demonstrated for the first time, are tuned using integrated electrowetting-on-dielectrics (EWOD) actuators. The router is assembled from two chips fabricated by standard MEMS techniques. Through a combination of microfluidic with micro-optical components on chip, reliable light routing is achieved with switching times of [1.5-3.3] s, efficiencies of coupling into channels of up to 12%, optical cross-talk as low as -24 dB, a required drive voltage of 50 V, and a low power consumption of <5 mW, using a device 12 × 13 × 2 mm(3) in size. The optofluidic approach enables addressing of multiple channels over a broad wavelength range. Such optical routing capabilities are important for lab-on-chip devices focusing on optical spectroscopy, optical detection, or even optical manipulation. When integrated with external light sources and a low-cost disposable photonic lab-on-a-chip, the router could thus lead to novel laboratory measurement systems.


Definition of bulky disease in early stage Hodgkin lymphoma in computed tomography era: prognostic significance of measurements in the coronal and transverse planes.
Disease bulk is an important prognostic factor in early stage Hodgkin lymphoma, but its definition is unclear in the computed tomography era. This retrospective analysis investigated the prognostic significance of bulky disease measured in transverse and coronal planes on computed tomography imaging. Early stage Hodgkin lymphoma patients (n=185) treated with chemotherapy with or without radiotherapy from 2000-2010 were included. The longest diameter of the largest lymph node mass was measured in transverse and coronal axes on pre-treatment imaging. The optimal cut off for disease bulk was maximal diameter greater than 7 cm measured in either the transverse or coronal plane. Thirty patients with maximal transverse diameter of 7 cm or under were found to have bulk in coronal axis. The 4-year overall survival was 96.5% (CI: 93.3%, 100%) and 4-year relapse-free survival was 86.8% (CI: 81.9%, 92.1%) for all patients. Relapse-free survival at four years for bulky patients was 80.5% (CI: 73%, 88.9%) compared to 94.4% (CI: 89.1%, 100%) for non-bulky; Cox HR 4.21 (CI: 1.43, 12.38) (P=0.004). In bulky patients, relapse-free survival was not impacted in patients treated with chemoradiotherapy; however, it was significantly lower in patients treated with chemotherapy alone. In an independent validation cohort of 38 patients treated with chemotherapy alone, patients with bulky disease had an inferior relapse-free survival [at 4 years, 71.1% (CI: 52.1%, 97%) vs 94.1% (CI: 83.6%, 100%), Cox HR 5.27 (CI: 0.62, 45.16); P=0.09]. Presence of bulky disease on multidimensional computed tomography imaging is a significant prognostic factor in early stage Hodgkin lymphoma. Coronal reformations may be included for routine Hodgkin lymphoma staging evaluation. In future, our definition of disease bulk may be useful in identifying patients who are most appropriate for chemotherapy alone.


The dystrophin gene and cognitive function in the general population.
The aim of our study is to investigate whether single-nucleotide dystrophin gene (DMD) variants associate with variability in cognitive functions in healthy populations. The study included 1240 participants from the Erasmus Rucphen family (ERF) study and 1464 individuals from the Rotterdam Study (RS). The participants whose exomes were sequenced and who were assessed for various cognitive traits were included in the analysis. To determine the association between DMD variants and cognitive ability, linear (mixed) modeling with adjustment for age, sex and education was used. Moreover, Sequence Kernel Association Test (SKAT) was used to test the overall association of the rare genetic variants present in the DMD with cognitive traits. Although no DMD variant surpassed the prespecified significance threshold (P<1 × 10(-4)), rs147546024:A>G showed strong association (β = 1.786, P-value = 2.56 × 10(-4)) with block-design test in the ERF study, while another variant rs1800273:G>A showed suggestive association (β = -0.465, P-value = 0.002) with Mini-Mental State Examination test in the RS. Both variants are highly conserved, although rs147546024:A>G is an intronic variant, whereas rs1800273:G>A is a missense variant in the DMD which has a predicted damaging effect on the protein. Further gene-based analysis of DMD revealed suggestive association (P-values = 0.087 and 0.074) with general cognitive ability in both cohorts. In conclusion, both single variant and gene-based analyses suggest the existence of variants in the DMD which may affect cognitive functioning in the general populations.


ORTHODONTIC RETENTION. Studies of retention capacity, cost-effectiveness and long-term stability.
Retention strategies, cost-effectiveness and long-term stability of treatment outcome are essential aspects of orthodontic treatment planning. The overall aim of this thesis was to compare and evaluate three different retention strategies, with special reference to short- and long-term clinical stability and cost-effectiveness. The approach was evidence-based, hence randomized controlled methodology was used in order to generate high levels of evidence. This thesis is based on four studies: Papers I and II are based on randomized controlled trials, evaluating the stability of treatment outcome after one and two years of retention, using three different retention strategies: a maxillary vacuum-formed retainer combined with a mandibular canine-to-canine retainer; a maxillary vacuum-formed retainer combined with stripping of the mandibular anterior teeth and a prefabricated positioner. Paper III presents a cost-minimization analysis of two years of retention treatment. Paper IV is based on a randomized controlled trial documenting the results five years post-retention. The following conclusions were drawn: Papers I and II. From a clinical perspective, asssessment after one year of retention disclosed that the three retention methods were successful in retaining the orthodontic treatment results. After two years of retention, all three retention methods were equally effective in controlling relapse at a clinically acceptable level. Most of the relapse occurred during the first year of retention; only minor or negligible changes were found during the second year. The subjects were grouped according to the level of compliance (excellent or good). After two years of retention there was a negative correlation between growth in body height and relapse of mandibular LII in the group of subjects with excellent compliance. The group with good compliance showed a positive correlation (Paper II, Figure 3). After two years of retention, growth in body height, initial crowding and gender had no significant influence on mandibular LII (Paper II, Figure 4 and Table 4). Paper III. The cost minimization analysis disclosed that although the three retention methods achieved clinically similar results, the associated societal costs differed. After two years of retention, the vacuum-formed retainer (VFR) in combination with a canine-to-canine retainer (CTC) was the least cost-effective retention appliance. Paper IV. After five years or more out of retention, the three retention methods had achieved equally favourable clinical results. Key conclusion and clinical implications. This study compared the short- and long-term outcomes of orthodontic retention by three different methods: a maxillary vacuum-formed retainer combined with a mandibular canine-to-canine retainer; a maxillary vacuum-formed retainer combined with stripping of the mandibular anterior teeth and a prefabricated positioner. All methods gave equally positive clinical results in both the short-term, i.e. after one and two years of retention, and in the long-term, five years or more post-retention. After two years of retention, the level of compliance affected the retention treatment result. However, no such effect was shown for body height, the severity of initial crowding or gender. Today, there is increasing emphasis on the importance of economic aspects of healthcare. Of the three methods evaluated in this study, the least cost-effective, after two years of retention, was a vacuum-formed retainer combined with a bonded canine-to-canine retainer. The clinical implication of this finding is that in patients meeting the inclusion criteria, interproximal stripping of the mandibular anterior teeth, or the use of a prefabricated positioner, are highly appropriate alternatives to a mandibular bonded canine-to-canine retainer. The overall conclusions are that there are a number of effective retention methods available and the clinician is not limited to routine use of a bonded mandibular canine-to-canine retainer. The most appropriate retention method should be selected on an individual, case to case basis, taking into account such variables as orthodontic diagnosis, the expected level of patient compliance, patient preferences and financial considerations.


Anchoring the human olfactory system within a functional gradient.
Margulies et al. (2016) demonstrated the existence of at least five independent functional connectivity gradients in the human brain. However, it is unclear how these functional gradients might link to anatomy. The dual origin theory proposes that differences in cortical cytoarchitecture originate from two trends of progressive differentiation between the different layers of the cortex, referred to as the hippocampocentric and olfactocentric systems. When conceptualising the functional connectivity gradients within the evolutionary framework of the Dual Origin theory, the first gradient likely represents the hippocampocentric system anatomically. Here we expand on this concept and demonstrate that the fifth gradient likely links to the olfactocentric system. We describe the anatomy of the latter as well as the evidence to support this hypothesis. Together, the first and fifth gradients might help to model the Dual Origin theory of the human brain and inform brain models and pathologies.


Shear stress in lattice Boltzmann simulations.
A thorough study of shear stress within the lattice Boltzmann method is provided. Via standard multiscale Chapman-Enskog expansion we investigate the dependence of the error in shear stress on grid resolution showing that the shear stress obtained by the lattice Boltzmann method is second-order accurate. This convergence, however, is usually spoiled by the boundary conditions. It is also investigated which value of the relaxation parameter minimizes the error. Furthermore, for simulations using velocity boundary conditions, an artificial mass increase is often observed. This is a consequence of the compressibility of the lattice Boltzmann fluid. We investigate this issue and derive an analytic expression for the time dependence of the fluid density in terms of the Reynolds number, Mach number, and a geometric factor for the case of a Poiseuille flow through a rectangular channel in three dimensions. Comparison of the analytic expression with results of lattice Boltzmann simulations shows excellent agreement.


Macroinvertebrate Taxonomic and Functional Trait Compositions within Lotic Habitats Affected By River Restoration Practices.
The widespread degradation of lotic ecosystems has prompted extensive river restoration efforts globally, but many studies have reported modest ecological responses to rehabilitation practices. The functional properties of biotic communities are rarely examined within post-project appraisals, which would provide more ecological information underpinning ecosystem responses to restoration practices and potentially pinpoint project limitations. This study examines macroinvertebrate community responses to three projects which aimed to physically restore channel morphologies. Taxonomic and functional trait compositions supported by widely occurring lotic habitats (biotopes) were examined across paired restored and non-restored (control) reaches. The multivariate location (average community composition) of taxonomic and functional trait compositions differed marginally between control and restored reaches. However, changes in the amount of multivariate dispersion were more robust and indicated greater ecological heterogeneity within restored reaches, particularly when considering functional trait compositions. Organic biotopes (macrophyte stands and macroalgae) occurred widely across all study sites and supported a high alpha (within-habitat) taxonomic diversity compared to mineralogical biotopes (sand and gravel patches), which were characteristic of restored reaches. However, mineralogical biotopes possessed a higher beta (between-habitat) functional diversity, although this was less pronounced for taxonomic compositions. This study demonstrates that examining the functional and structural properties of taxa across distinct biotopes can provide a greater understanding of biotic responses to river restoration works. Such information could be used to better understand the ecological implications of rehabilitation practices and guide more effective management strategies.


Targeted disruption of SMAD3 results in impaired mucosal immunity and diminished T cell responsiveness to TGF-beta.
SMAD3 is one of the intracellular mediators that transduces signals from transforming growth factor-beta (TGF-beta) and activin receptors. We show that SMAD3 mutant mice generated by gene targeting die between 1 and 8 months due to a primary defect in immune function. Symptomatic mice exhibit thymic involution, enlarged lymph nodes, and formation of bacterial abscesses adjacent to mucosal surfaces. Mutant T cells exhibit an activated phenotype in vivo, and are not inhibited by TGF-beta1 in vitro. Mutant neutrophils are also impaired in their chemotactic response toward TGF-beta. Chronic intestinal inflammation is infrequently associated with colonic adenocarcinoma in mice older than 6 months of age. These data suggest that SMAD3 has an important role in TGF-beta-mediated regulation of T cell activation and mucosal immunity, and that the loss of these functions is responsible for chronic infection and the lethality of Smad3-null mice.


Accumulation of tissue factor into developing thrombi in vivo is dependent upon microparticle P-selectin glycoprotein ligand 1 and platelet P-selectin.
Using a laser-induced endothelial injury model, we examined thrombus formation in the microcirculation of wild-type and genetically altered mice by real-time in vivo microscopy to analyze this complex physiologic process in a system that includes the vessel wall, the presence of flowing blood, and the absence of anticoagulants. We observe P-selectin expression, tissue factor accumulation, and fibrin generation after platelet localization in the developing thrombus in arterioles of wild-type mice. However, mice lacking P-selectin glycoprotein ligand 1 (PSGL-1) or P-selectin, or wild-type mice infused with blocking P-selectin antibodies, developed platelet thrombi containing minimal tissue factor and fibrin. To explore the delivery of tissue factor into a developing thrombus, we identified monocyte-derived microparticles in human platelet-poor plasma that express tissue factor, PSGL-1, and CD14. Fluorescently labeled mouse microparticles infused into a recipient mouse localized within the developing thrombus, indicating that one pathway for the initiation of blood coagulation in vivo involves the accumulation of tissue factor- and PSGL-1-containing microparticles in the platelet thrombus expressing P-selectin. These monocyte-derived microparticles bind to activated platelets in an interaction mediated by platelet P-selectin and microparticle PSGL-1. We propose that PSGL-1 plays a role in blood coagulation in addition to its known role in leukocyte trafficking.


Accelerated treatment of postpneumonectomy empyema: a binational long-term study.
Postpneumonectomy empyema remains a clinical challenge. We proposed an accelerated therapy without an open chest window 5 years ago. This concept was evaluated on a larger scale in 2 centers in 2 different countries. Between July 1995 and October 2005, 75 consecutive patients with postpneumonectomy empyema were treated in Szczecin, Poland (n = 35), and Zurich, Switzerland (n = 40). The therapy consisted of repeated open surgical debridement of the pleural cavity after achievement of general anesthesia, a negative pressure wound therapy of the temporarily closed chest cavity filled with povidone-iodine-soaked towels, and continuous suction and systemic antimicrobial therapy. If present, bronchopleural fistulae were closed and reinforced either with a muscle flap or the omentum. Finally, the pleural space was filled with an antibiotic solution and definitively closed. Of 75 patients (63 men; median age, 59 years; age range, 19-82 years), postpneumonectomy empyema was present on the right in 46 patients (32 with bronchopleural fistula) and in 29 patients (12 with bronchopleural fistula) on the left. Median time between pneumonectomy and postpneumonectomy empyema was 131 days (range, 7-7200 days). Bronchopleural fistulae have been closed and additionally reinforced by means of different methods (omentum, 18; muscle, 11; pericardial fat, 5; azygos vein, 1). The chest was definitively closed within 8 days in 94.6% of patients. The median hospitalization time was 18 days (range, 9-134 days). Postpneumonectomy empyema was successfully treated in 97.3% of patients, including 10 (13%) patients who needed a second treatment cycle. Three (4%) patients died within 90 days. The median follow-up time was 29.5 moths (range, 3-107 months). Treatment of postpneumonectomy empyema with the accelerated treatment is effective and safe. Our results are superior compared with those in reported series using a (temporary) chest fenestration. Patients appreciate the physical integrity of the chest.


Extremely metal-poor stars from the cosmic dawn in the bulge of the Milky Way.
The first stars are predicted to have formed within 200 million years after the Big Bang, initiating the cosmic dawn. A true first star has not yet been discovered, although stars with tiny amounts of elements heavier than helium ('metals') have been found in the outer regions ('halo') of the Milky Way. The first stars and their immediate successors should, however, preferentially be found today in the central regions ('bulges') of galaxies, because they formed in the largest over-densities that grew gravitationally with time. The Milky Way bulge underwent a rapid chemical enrichment during the first 1-2 billion years, leading to a dearth of early, metal-poor stars. Here we report observations of extremely metal-poor stars in the Milky Way bulge, including one star with an iron abundance about 10,000 times lower than the solar value without noticeable carbon enhancement. We confirm that most of the metal-poor bulge stars are on tight orbits around the Galactic Centre, rather than being halo stars passing through the bulge, as expected for stars formed at redshifts greater than 15. Their chemical compositions are in general similar to typical halo stars of the same metallicity although intriguing differences exist, including lower abundances of carbon.


Barrier dysfunction or drainage reduction: differentiating causes of CSF protein increase.
Cerebrospinal fluid (CSF) protein analysis is an important element in the diagnostic chain for various central nervous system (CNS) pathologies. Among multiple existing approaches to interpreting measured protein levels, the Reiber diagram is particularly robust with respect to physiologic inter-individual variability, as it uses multiple subject-specific anchoring values. Beyond reliable identification of abnormal protein levels, the Reiber diagram has the potential to elucidate their pathophysiologic origin. In particular, both reduction of CSF drainage from the cranio-spinal space as well as blood-CNS barrier dysfunction have been suggested ρas possible causes of increased concentration of blood-derived proteins. However, there is disagreement on which of the two is the true cause. We designed two computational models to investigate the mechanisms governing protein distribution in the spinal CSF. With a one-dimensional model, we evaluated the distribution of albumin and immunoglobulin G (IgG), accounting for protein transport rates across blood-CNS barriers, CSF dynamics (including both dispersion induced by CSF pulsations and advection by mean CSF flow) and CSF drainage. Dispersion coefficients were determined a priori by computing the axisymmetric three-dimensional CSF dynamics and solute transport in a representative segment of the spinal canal. Our models reproduce the empirically determined hyperbolic relation between albumin and IgG quotients. They indicate that variation in CSF drainage would yield a linear rather than the expected hyperbolic profile. In contrast, modelled barrier dysfunction reproduces the experimentally observed relation. High levels of albumin identified in the Reiber diagram are more likely to originate from a barrier dysfunction than from a reduction in CSF drainage. Our in silico experiments further support the hypothesis of decreasing spinal CSF drainage in rostro-caudal direction and emphasize the physiological importance of pulsation-driven dispersion for the transport of large molecules in the CSF.


Adverse neurodevelopmental outcomes among extremely low birth weight infants with a normal head ultrasound: prevalence and antecedents.
Severe abnormalities of the head ultrasound (HUS) are important predictors of cerebral palsy (CP) and mental retardation, and a normal HUS usually ensures the absence of major impairments. With the increasing survival of extremely low birth weight (ELBW) infants (birth weight <1000 g), the prognostic significance of a normal HUS may differ. This study examined the prevalence of and risk factors for CP and impaired mental development among ELBW infants with a normal HUS. Study infants were ELBW infants who were cared for in Neonatal Research Network centers in the years 1995-1999, had a normal early and late HUS, survived to discharge, and returned for follow-up assessments at 18 to 22 months' corrected age. The outcomes of interest were a score <70 on the Bayley Scales of Infant Development-II Mental Developmental Index (MDI) and CP. Risk factors included maternal demographics; infant characteristics; and interventions or morbidities related to the lung, infection, and nutrition. Logistic regression was used to estimate odds ratios (ORs) and 95% confidence intervals (CIs). A time-oriented approach was used to select variables for inclusion in logistic models. Of 1749 infants with a normal early and late HUS (performed at a mean age of 6 and 47 days, respectively), 1473 (84%) returned for follow-up assessment. Infants had a birth weight of 792 +/- 134 g (mean +/- SD) and gestational age of 26 +/- 2 weeks. Rates of CP and MDI <70 were 9.4% and 25.3%, respectively, and 29.2% of infants had either CP or MDI <70. In multivariate analyses, factors associated with CP were male gender (OR: 1.8; 95% CI: 1.2-2.6), multiple birth, (OR: 1.6; 95% CI: 1.1-2.5), decreasing birth weight (OR: 1.3 for each 100-g decrease; 95% CI: 1.1-1.5), pneumothorax (OR: 2.3; 95% CI: 1.2-4.4), and days of conventional ventilation (OR: 1.2 for each additional 10 days; 95% CI: 1.1-1.3). With the exception of pneumothorax, these same factors were associated with MDI <70, in addition to less maternal education (OR: 1.4; 95% CI: 1.0-1.9) and Medicaid or lack of coverage for maternal insurance (OR: 1.7; 95% CI: 1.2-2.4). Nearly 30% of ELBW infants with a normal HUS had either CP or a low MDI. Risk factors that are associated with this high rate of adverse outcomes include pneumothorax, prolonged exposure to mechanical ventilation, and educational and economic disadvantage. Improvements in pulmonary care to reduce duration of ventilation and avoid air leaks might improve neurodevelopmental outcome for ELBW infants.


[Adherence to institutional clinical practice guideline on managing upper gastrointestinal bleeding, in a hospital in Lima - Peru].
In 2018, the Clinical Practice Guideline (CPG) for the Evaluation and Management of upper gastrointestinal bleeding (UGB) was published by the Social Security of Peru (EsSalud). It provides evidence-based statements to optimize the management of these patients. To evaluate the adherence to the statements of the CPG at the Edgardo Rebagliati Martins National Hospital (HNERM) of EsSalud (Lima, Peru). Retrospective study, which analyzed the database of all patients who came to the HNERM emergency service with suspected UGB and were scheduled for endoscopy between October 19, 2019 and April 15, 2020. We included those with ≥ 18 years of age. This database contains the main characteristics of the standardized medical history for patients with UGB. Compliance with 13 of the 34 statements of the EsSalud CPG was evaluated. The results were presented descriptively, and the factors associated with compliance with the statements with insufficient adherence (<80%) and with more than 100 evaluated participants were evaluated. Data were obtained from 184 patients who met the inclusion criteria (men: 59.8%, median age: 70 years). The range of adherence to the 13 statements was from 63.2% to 99.5%. Only two statements had insufficient adherence (<80%). The statement with the least adherence was the recommendation to perform a restrictive transfusion. Noncompliance with this recommendation was found to be lower in those who had a higher score on the Glasgow-Blatchford index, a urea creatinine ratio > 60, and a lower hemoglobin on admission. Of the 13 statements evaluated, 11 had satisfactory adherence. It is important to explore the reasons why adherence is not adequate for some statements, and to evaluate methods to increase this adherence.


Outdoor Air Pollution and New-Onset Airway Disease. An Official American Thoracic Society Workshop Report.
Although it is well accepted that air pollution exposure exacerbates preexisting airway disease, it has not been firmly established that long-term pollution exposure increases the risk of new-onset asthma or chronic obstruction pulmonary disease (COPD). This Workshop brought together experts on mechanistic, epidemiological, and clinical aspects of airway disease to review current knowledge regarding whether air pollution is a causal factor in the development of asthma and/or COPD. Speakers presented recent evidence in their respective areas of expertise related to air pollution and new airway disease incidence, followed by interactive discussions. A writing committee summarized their collective findings. The Epidemiology Group found that long-term exposure to air pollution, especially metrics of traffic-related air pollution such as nitrogen dioxide and black carbon, is associated with onset of childhood asthma. However, the evidence for a causal role in adult-onset asthma or COPD remains insufficient. The Mechanistic Group concluded that air pollution exposure can cause airway remodeling, which can lead to asthma or COPD, as well as asthma-like phenotypes that worsen with long-term exposure to air pollution, especially fine particulate matter and ozone. The Clinical Group concluded that air pollution is a plausible contributor to the onset of both asthma and COPD. Available evidence indicates that long-term exposure to air pollution is a cause of childhood asthma, but the evidence for a similar determination for adult asthma or COPD remains insufficient. Further research is needed to elucidate the exact biological mechanism underlying incident childhood asthma, and the specific air pollutant that causes it.