The Eukaryotic Host Factor 14-3-3 Inactivates Adenylate Cyclase Toxins of Bordetella bronchiseptica and B. parapertussis, but Not B. pertussis.
Bordetella pertussis, Bordetella bronchiseptica, and Bordetella parapertussis share highly homologous virulence factors and commonly cause respiratory infections in mammals; however, their host specificities and disease severities differ, and the reasons for this remain largely unknown. Adenylate cyclase toxin (CyaA) is a homologous virulence factor that is thought to play crucial roles in Bordetella infections. We herein demonstrate that CyaAs function as virulence factors differently between B. bronchiseptica/B. parapertussis and B. pertussisBbronchiseptica CyaA bound to target cells, and its enzyme domain was translocated into the cytosol similarly to Bpertussis CyaA. The hemolytic activity of Bbronchiseptica CyaA on sheep erythrocytes was also preserved. However, in nucleated target cells, Bbronchiseptica CyaA was phosphorylated at Ser375, which constitutes a motif (RSXpSXP [pS is phosphoserine]) recognized by the host factor 14-3-3, resulting in the abrogation of adenylate cyclase activity. Consequently, the cytotoxic effects of Bbronchiseptica CyaA based on its enzyme activity were markedly attenuated. Bparapertussis CyaA carries the 14-3-3 motif, indicating that its intracellular enzyme activity is abrogated similarly to Bbronchiseptica CyaA; however, Bpertussis CyaA has Phe375 instead of Ser, and thus, was not affected by 14-3-3. In addition, Bpertussis CyaA impaired the barrier function of epithelial cells, whereas Bbronchiseptica CyaA did not. Rat infection experiments suggested that functional differences in CyaA are related to differences in pathogenicity between B. bronchiseptica/Bparapertussis and B. pertussisIMPORTANCEBordetella pertussis, B. bronchiseptica, and B. parapertussis are bacterial respiratory pathogens that are genetically close to each other and produce many homologous virulence factors; however, their host specificities and disease severities differ, and the reasons for this remain unknown. Previous studies attempted to explain these differences by the distinct virulence factors produced by each Bordetella species. In contrast, we indicated functional differences in adenylate cyclase toxin, a homologous virulence factor of Bordetella The toxins of B. bronchiseptica and presumably B. parapertussis were inactivated by the host factor 14-3-3 after phosphorylation in target cells, whereas the B. pertussis toxin was not inactivated because of the lack of the phosphorylation site. This is the first study to show that 14-3-3 inactivates the virulence factors of pathogens. The present results suggest that pathogenic differences in Bordetella are attributed to the different activities of adenylate cyclase toxins.


Modelling the electrical properties of bladder tissue--quantifying impedance changes due to inflammation and oedema.
Electrical impedance spectroscopy has been developed as a potential method for the diagnosis of carcinoma in epithelial tissues. An understanding of the influence of structural changes in the tissue on the properties measured using this technique is essential for interpreting measured data and optimization of probe design. In contrast to other tissue types, carcinoma in situ of the bladder gives rise to an increase in electrical impedance over the kHz-MHz frequency range in comparison to normal tissue. Finite element models of the urothelium and the underlying superficial lamina propria have been constructed and solved in order to ascertain the influence of structural changes associated with malignancy, oedema and inflammation on the measured electrical properties of the tissue. Sensitivity analysis of results from a composite tissue model suggests that the increase in lymphocyte density in the lamina propria associated with an inflammatory response to the infiltration of urine into the tissue may explain these unusual electrical properties.


Protective efficacy of Emblica officinalis against Klebsiella pneumoniae induced pneumonia in mice.
Emblica officinalis (amla), which is a good source of vitamin C, has been shown to be beneficial due to its immune system enhancing property coupled with its tonifying and antiageing effect. The present study was conducted to evaluate the effect of E. officinalis feeding on the susceptibility of experimental mice to respiratory tract infection induced by Klebsiella pneumoniae. The effect of short- (15 days) and long (30 days)-term feeding of amla in mice on the course of K. pneumoniae ATCC43816 infection in lungs was studied, in terms of bacterial colonization, macrophage activity, malondialdehyde (MDA) and nitrite production in broncheoalveolar lavage fluid (BALF). Tumour necrosis factor (TNF)-alpha level in serum was also assessed. Though there was a decrease in bacterial colonization after short-term feeding, it was not significant. On the contrary, the decrease in bacterial load was significant (P < 0.05) on long-term feeding. The operative mechanisms in terms of lipid peroxidation, phagocytosis and nitrite production were studied by estimating their levels in broncheoalveolar lavage fluid (BALF). Maximum decrease in malondialdehyde (MDA) levels and increase in phagocytic activity and nitrite levels on long-term feeding was seen. These results suggest that dietary supplementation with amla protects against bacterial colonization of lungs on long-term feeding in experimental model. Further studies need to be conducted to understand the actual mechanism.


Life-long consequences of postnatal normoxia exposure in rats raised at high altitude.
We tested the hypothesis that exposure of high-altitude (HA) rats to a period of postnatal normoxia has long-term consequences on the ventilatory and hematological acclimatization in adults. Male and female HA rats (3,600 m, Po(2) ≃ 100 Torr; La Paz, Bolivia) were exposed to normal room air [HA control (HACont)] or enriched oxygen (32% O(2); Po(2) ≃ 160 Torr) from 1 day before to 15 days after birth [HA postnatal normoxia (HApNorm)]. Hematocrit and hemoglobin values were assessed at 2, 12, and 32 wk of age. Cardiac and lung morphology were assessed at 12 wk by measuring right ventricular hypertrophy (pulmonary hypertension index) and lung air space-to-tissue ratio (indicative of alveolarization). Respiratory parameters under baseline conditions and in response to 32% O(2) for 10 min (relieving the ambient hypoxic stimulus) were measured by whole body plethysmography at 12 wk. Finally, we performed a survival analysis up to 600 days of age. Compared with HACont, HApNorm rats had reduced hematocrit and hemoglobin levels at all ages (both sexes); reduced right ventricular hypertrophy (both sexes); lower air space-to-tissue ratio in the lungs (males only); reduced CO(2) production rate, but higher oxygen uptake (males only); and similar respiratory frequency, tidal volume, and minute ventilation. When breathing 32% O(2), HApNorm male rats had a stronger decrease of minute ventilation than HACont. HApNorm rats had a marked tendency toward longer survival throughout the study. We conclude that exposure to ambient hypoxia during postnatal development in HA rats has deleterious consequences on acclimatization to hypoxia as adults.


Ergonomic hazards in otolaryngology.
To evaluate the presence of postural-related strain and musculoskeletal discomfort, along with the level of ergonomics training and the availability of ergonomic equipment among otolaryngology surgeons. Intraoperative observations and survey study. Using the Rapid Entire Body Assessment score system to identify ergonomic hazards, we conducted intraoperative observations assessing operating room personnel during different otolaryngological subspecialty procedures. Based on these findings, otolaryngology surgeons at a single academic institution in the United States were sent a survey that evaluated ergonomic practice, environmental infrastructure, and prior ergonomic training or education. A response rate of 69% was obtained from 70 surgeons, with 72.9% of responding surgeons suffering from some level of back pain, with cervical spine pain being the most common. Interestingly, residents were equally affected when compared to more senior surgeons both in subjective survey reports and from observational risk analysis. Furthermore, 43.8% of surgeons reported suffering from the highest level of pain when standing, whereas only 12.5% experienced pain when sitting. Importantly, 10% stated that pain impacted their work. Only 24% of surgeons had any prior ergonomic training or education. Our data suggest that pain and disability induced by poor ergonomics are widespread among the otolaryngology community and confirm that surgeons rarely receive ergonomic training in the surgical context. Additionally, intraoperative observational findings identified that the majority of observed surgeons display poor posture, particularly a poor cervical angle and use of ergonomic setups, both of which increase ergonomic risk hazard. These data provide guidance for future interventional studies. NA Laryngoscope, 129:370-376, 2019.


Modulation of in vivo GABA-evoked responses by nitric oxide-active compounds in the globus pallidus of rat.
Nitric oxide (NO) is a gaseous molecule acting as a messenger in both the peripheral and the central nervous systems. NO affects synaptic activity by modulating neurotransmitter release and/or receptor function. We previously observed that NO-active compounds modify the bioelectric activity of basal ganglia (BG) units. In this study, we applied microiontophoresis to extracellular in vivo recordings to investigate the effect of NO-active compounds on GABA-evoked responses in the globus pallidus (GP) of anesthetized rats. The changes induced by NO-active drugs on the GABA-induced inhibition were used as indicators of NO modulation. The response to GABA release was tested on recorded GP neurons before and during the administration of S-nitroso-glutathione (SNOG, a NO donor) and/or Nω-nitro-L: -arginine methyl ester (L: -NAME), an inhibitor of nitric oxide synthase (NOS); furthermore, SNOG and L: -NAME were tested at different ejection currents in order to highlight the possibility of a current-dependent effect in the nitrergic modulation of GABA transmission. In general, during SNOG ejection the magnitude of GABA-evoked responses was reduced, whereas the administration of L: -NAME produced the opposite effect. The results suggest that NO-active drugs modulate the response of GP neurons to GABA transmission; the effects induced by SNOG and L: -NAME were strictly related to the ejection currents. Then, the modulation of GABAergic transmission by NO could represent a mechanism to finely regulate the GP neurons activity with important consequences on the overall BG function.


Participation profiles in domestic life and peer relations as experienced by adolescents with and without impairments and long-term health conditions.
To investigate how individual and environmental factors relate to self-reported participation profiles in adolescents with and without impairments or long-term health conditions. A person-oriented approach (hierarchical cluster analysis) was used to identify cluster groups of individuals sharing participation patterns in the outcome variables frequency perceived importance in domestic life and peer relations. Cluster groups were compared using one-way analysis of variance (ANOVA). A nine-cluster solution was chosen. All clusters included adolescents with impairment and long-term health conditions. Perceived importance of peer relations was more important than frequent attendance in domestic-life activities. Frequency of participation in dialogues and family interaction patterns seemed to affect the participation profiles more than factors related to body functions. Type of impairment or long-term health condition is a weaker determinant of membership in clusters depicting frequency and perceived importance in domestic life or peer relations than dialogue and family environment.


A property-based analysis of human transcription factors.
Transcription factors are essential proteins for regulating gene expression. This regulation depends upon specific features of the transcription factors, including how they interact with DNA, how they interact with each other, and how they are post-translationally modified. Reliable information about key properties associated with transcription factors will therefore be useful for data analysis, in particular of data from high-throughput experiments. We have used an existing list of 1978 human proteins described as transcription factors to make a well-annotated data set, which includes information on Pfam domains, DNA-binding domains, post-translational modifications and protein-protein interactions. We have then used this data set for enrichment analysis. We have investigated correlations within this set of features, and between the features and more general protein properties. We have also used the data set to analyze previously published gene lists associated with cell differentiation, cancer, and tissue distribution. The study shows that well-annotated feature list for transcription factors is a useful resource for extensive data analysis; both of transcription factor properties in general and of properties associated with specific processes. However, the study also shows that such analyses are easily biased by incomplete coverage in experimental data, and by how gene sets are defined.


Anti-inflammatory Activity of Extra Virgin Olive Oil Polyphenols: Which Role in the Prevention and Treatment of Immune-Mediated Inflammatory Diseases?
Altered inflammatory response characterizes chronic immunemediated inflammatory diseases (IMID) such as rheumatoid arthritis, inflammatory bowel disease, multiple sclerosis, systemic lupus erythematosus, and psoriasis. Accumulating evidence indicates that regular consumption of extra virgin olive oil (EVOO), the main source of fat in the Mediterranean diet, is associated with a reduced risk of developing chronic degenerative disorders such as cardiovascular diseases, type 2 diabetes and cancer. The beneficial effects on health of EVOO have been attributed, besides to the monounsaturated fats content, to the presence of phenolic compounds that have antioxidant, anti-inflammatory and immunomodulatory properties. The purpose of this review is to provide an overview of the effects of EVOO polyphenols on IMID highlighting the potential mechanisms of action. Scientific papers were found by searching in PubMed up to May 2017 using the following key words: rheumatoid arthritis, inflammatory bowel disease, multiple sclerosis, systemic lupus erythematosus, and psoriasis also in combination with EVOO, phenolic compounds, oleuropein, oleocantal, hydroxytyrosol,tyrosol and oleochantal. In vitro and in vivo studies indicate that EVOO and its polyphenols can improve diseases symptoms in IMID, by acting both at local and systemic levels and by modulating several molecular pathways. Nevertheless, there are not sufficient data to achieve specific nutritional guidelines. Further research is needed to evaluate the real contribution of EVOO and its phenolic compounds in modulating the IMID-associated inflammatory perturbations, in order to develop appropriate nutritional recommendations.


Parallel detection algorithm for page-oriented optical memories.
We present a parallel algorithm for the reliable detection of two-dimensional binary data in page-oriented memories. The development of the proposed pseudodecision-feedback equalization (PDFE) method is motivated by the classical decision-feedback equalization receiver. The technique takes advantage of the known or the estimated optical system characteristics to mitigate space-variant blur and additive thermal noise. We extend the method to correct for fixed-pattern errors including magnification, rotation, and transverse shift. Advantages of the PDFE algorithm include its parallel design, low computational complexity, and local connectivity. A system-capacity metric is used to compare the performance of the PDFE receiver with other conventional approaches, including the simple threshold, the 1:2 modulation code, and the Wiener filter. Results show the PDFE to outperform all the above techniques over a variety of channels for both incoherent and coherent systems. Implementation issues are discussed, and a MOSIS (Metal-Oxide Semiconductor Implementation Service) 2-mum design is presented.


Escherichia coli adenylate kinase dynamics: comparison of elastic network model modes with mode-coupling (15)N-NMR relaxation data.
The dynamics of adenylate kinase of Escherichia coli (AKeco) and its complex with the inhibitor AP(5)A, are characterized by correlating the theoretical results obtained with the Gaussian Network Model (GNM) and the anisotropic network model (ANM) with the order parameters and correlation times obtained with Slowly Relaxing Local Structure (SRLS) analysis of (15)N-NMR relaxation data. The AMPbd and LID domains of AKeco execute in solution large amplitude motions associated with the catalytic reaction Mg(+2)*ATP + AMP --> Mg(+2)*ADP + ADP. Two sets of correlation times and order parameters were determined by NMR/SRLS for AKeco, attributed to slow (nanoseconds) motions with correlation time tau( perpendicular) and low order parameters, and fast (picoseconds) motions with correlation time tau( parallel) and high order parameters. The structural connotation of these patterns is examined herein by subjecting AKeco and AKeco*AP(5)A to GNM analysis, which yields the dynamic spectrum in terms of slow and fast modes. The low/high NMR order parameters correlate with the slow/fast modes of the backbone elucidated with GNM. Likewise, tau( parallel) and tau( perpendicular) are associated with fast and slow GNM modes, respectively. Catalysis-related domain motion of AMPbd and LID in AKeco, occurring per NMR with correlation time tau( perpendicular), is associated with the first and second collective slow (global) GNM modes. The ANM-predicted deformations of the unliganded enzyme conform to the functional reconfiguration induced by ligand-binding, indicating the structural disposition (or potential) of the enzyme to bind its substrates. It is shown that NMR/SRLS and GNM/ANM analyses can be advantageously synthesized to provide insights into the molecular mechanisms that control biological function.


Stochastic simulations of minimal cells: the Ribocell model.
Over the last two decades, lipid compartments (liposomes, lipid-coated droplets) have been extensively used as in vitro "minimal" cell models. In particular, simple and complex biomolecular reactions have been carried out inside these self-assembled micro- and nano-sized compartments, leading to the synthesis of RNA and functional proteins inside liposomes. Despite this experimental progress, a detailed physical understanding of the underlying dynamics is missing. In particular, the combination of solute compartmentalization, reactivity and stochastic effects has not yet been clarified. A combination of experimental and computational approaches can reveal interesting mechanisms governing the behavior of micro compartmentalized systems, in particular by highlighting the intrinsic stochastic diversity within a population of "synthetic cells". In this context, we have developed a computational platform called ENVIRONMENT suitable for studying the stochastic time evolution of reacting lipid compartments. This software - which implements a Gillespie Algorithm - is an improvement over a previous program that simulated the stochastic time evolution of homogeneous, fixed-volume, chemically reacting systems, extending it to more general conditions in which a collection of similar such systems interact and change over the course of time. In particular, our approach is focused on elucidating the role of randomness in the time behavior of chemically reacting lipid compartments, such as micelles, vesicles or micro emulsions, in regimes where random fluctuations due to the stochastic nature of reacting events can lead an open system towards unexpected time evolutions. This paper analyses the so-called Ribocell (RNA-based cell) model. It consists in a hypothetical minimal cell based on a self-replicating minimum RNA genome coupled with a self-reproducing lipid vesicle compartment. This model assumes the existence of two ribozymes, one able to catalyze the conversion of molecular precursors into lipids and the second able to replicate RNA strands. The aim of this contribution is to explore the feasibility of this hypothetical minimal cell. By deterministic kinetic analysis, the best external conditions to observe synchronization between genome self-replication and vesicle membrane reproduction are determined, while its robustness to random fluctuations is investigated using stochastic simulations, and then discussed.


Motor cortex stimulation for the enhancement of recovery from stroke: a prospective, multicenter safety study.
Functional magnetic resonance imaging and transcranial magnetic stimulation studies suggest that human cortex shows evidence of neuroplasticity. Preclinical studies in rats and monkeys suggest that motor cortical stimulation can enhance plasticity and improve recovery after stroke. This study assesses the safety and preliminary efficacy of targeted subthreshold epidural cortical stimulation delivered concurrently with intensive rehabilitation therapy while using an investigational device in patients with chronic hemiparetic stroke. This is a prospective, multicenter, and nonblinded trial randomizing patients to rehabilitation with or without cortical stimulation. Patients aged 20 to 75 years who had had an ischemic stroke at least 4 months previously causing persistent moderate weakness of the arm were included. Functional magnetic resonance imaging localized hand motor function before surgery to place an epidural cortical electrode. Both groups then underwent rehabilitation for 3 weeks after which the electrode was removed. Outcome measures were obtained at baseline, during therapy, and at 1, 4, 8, and 12 weeks postprocedure. Ten patients were randomized; six patients to surgery, four to the control group. No patient deaths, neurological deterioration, or seizures occurred. There were two infections from nonprotocol-related causes. Of the eight patients completing the treatment, the stimulation plus rehabilitation group improved significantly better than controls in the Upper Extremity Fugl-Meyer (P = 0.003 overall) and the hand function score of the Stroke Impact Scale (P = 0.001 overall). The technique of cortical stimulation to enhance stroke recovery is well tolerated and safe.


Conformational analysis of receptor selective tachykinin analogs: senktide and septide.
The conformational behavior in solution of two receptor selective tachykinin agonists, senktide (succinyl-D-F-MeF-G-L-M-NH2) and septide (pQ-F-F-P-L-M-NH2) is described. Two dimensional cross relaxation NMR spectroscopy is used together with coupling constant data to obtain interproton distance constraints. These results are used in conjunction with semi-empirical energy computations to indicate favorable conformations. Senktide is found to have a high degree of conformational order which is attributed to rotational restriction associated with the N-methylation of phenylalanine. The lowest energy conformation in accord with the experimental interproton distances contains a beta-turn. Interproton distances indicate that septide exists as a random coil or in an extended chain conformation. Energy computations suggest that septide is primarily an extended chain with internal reorientation restricted by the proline residue. These results may be related to the selectivity of these peptides for different receptors, in that the analogs, with conformations more stable than tachykinins, are more receptor selective.


Similar Running Economy With Different Running Patterns Along the Aerial-Terrestrial Continuum.
No unique or ideal running pattern is the most economical for all runners. Classifying the global running patterns of individuals into 2 categories (aerial and terrestrial) using the Volodalen method could permit a better understanding of the relationship between running economy (RE) and biomechanics. The main purpose was to compare the RE of aerial and terrestrial runners. Two coaches classified 58 runners into aerial (n = 29) or terrestrial (n = 29) running patterns on the basis of visual observations. RE, muscle activity, kinematics, and spatiotemporal parameters of both groups were measured during a 5-min run at 12 km/h on a treadmill. Maximal oxygen uptake (V̇O2max) and peak treadmill speed (PTS) were assessed during an incremental running test. No differences were observed between aerial and terrestrial patterns for RE, V̇O2max, and PTS. However, at 12 km/h, aerial runners exhibited earlier gastrocnemius lateralis activation in preparation for contact, less dorsiflexion at ground contact, higher coactivation indexes, and greater leg stiffness during stance phase than terrestrial runners. Terrestrial runners had more pronounced semitendinosus activation at the start and end of the running cycle, shorter flight time, greater leg compression, and a more rear-foot strike. Different running patterns were associated with similar RE. Aerial runners appear to rely more on elastic energy utilization with a rapid eccentric-concentric coupling time, whereas terrestrial runners appear to propel the body more forward rather than upward to limit work against gravity. Excluding runners with a mixed running pattern from analyses did not affect study interpretation.


Activity-Dependent Myelination of Parvalbumin Interneurons Mediated by Axonal Morphological Plasticity.
Axonal myelination of neocortical pyramidal neurons is modulated dynamically by neuronal activity. Recent studies have shown that a substantial proportion of neocortical myelin content is contributed by fast-spiking, parvalbumin (PV)-positive interneurons. However, it remains unknown whether the myelination of PV+ interneurons is also modulated by intrinsic activity. Here, we used cell-type-specific Designer Receptors Exclusively Activated by Designer Drugs (DREADDs) in adult mice to activate a sparse population of medial prefrontal cortex (mPFC) PV+ interneurons. Using single-cell axonal reconstructions, we found that DREADD-stimulated PV+ interneurons exhibited a nearly two-fold increase in total length of myelination, predominantly mediated by a parallel increase of axonal arborization and number of internodes. In contrast, the distribution of axonal interbranch segment distance and myelin internode length were not altered significantly. Topographical analysis revealed that myelination of DREADD-stimulated cells extended to higher axonal branch orders while retaining a similar interbranch distance threshold for myelination. Together, our results demonstrate that chemogenetically induced neuronal activity increases the myelination of neocortical PV+ interneurons mediated at least in part by an elaboration of their axonal morphology.SIGNIFICANCE STATEMENT Myelination is the wrapping of an axon to optimize conduction velocity in an energy-efficient manner. Previous studies have shown that myelination of neocortical pyramidal neurons is experience and activity dependent. We now show that activity-dependent myelin plasticity in the adult neocortex extends to parvalbumin (PV)-expressing fast-spiking interneurons. Chemogenetic stimulation of PV interneurons in the medial prefrontal cortex (mPFC) significantly enhanced axonal myelination, which was paralleled by an increase in axonal arborization. This suggests that activity-dependent axonal plasticity may involve changes in both structural morphology and myelination. Such multicomponent plasticity reveals an unexpected repertoire of anatomical parameters available for optimizing and adapting neuronal networks in response to experience.


Exploring highly correlated materials via electron pair emission: the case of NiO/Ag(100).
Metal oxides like NiO are usually termed 'highly correlated', because the material properties are decisively determined by the electron-electron interaction. This makes them interesting candidates for electron pair spectroscopy which is particularly sensitive to the electron correlation. We have prepared ultrathin NiO/Ag(100) films and studied the electron pair emission upon electron impact. Compared to the metal substrate we observe an increase of the coincidence intensity by a factor of 8 for NiO. Thickness dependent measurements prove that this enhancement is an intrinsic effect rather than due to a mean free path increase of the oxide. The Néel temperature T(N) of NiO films displays a thickness dependence which allows us to tune T(N). We performed temperature dependent measurements and observed no temperature dependence of the coincidence spectra. This proves that the electron pair emission probes the local correlation rather than long range order. An enhanced coincidence intensity was also found for other oxide phases compared to their corresponding metal phases.


Factors affecting mobilization of peripheral blood progenitor cells in patients with lymphoma.
The objective of this study was to identify factors associated with poor mobilization of peripheral blood progenitor cells (PBPCs) or delayed platelet engraftment after high-dose therapy and autologous stem cell transplantation in patients with lymphoma. Fifty-eight patients with Hodgkin's disease or non-Hodgkin's lymphoma underwent PBPC transplantation as the "best available therapy" at Memorial Sloan-Kettering Cancer Center (New York, NY) between 1993 and 1995. PBPCs were mobilized with either granulocyte colony-stimulating factor (G-CSF) alone (n = 19) or G-CSF following combination chemotherapy (n = 39). Forty-eight of these patients underwent a PBPC transplant, receiving a conditioning regimen containing cyclophosphamide, etoposide, and either total body irradiation, total lymphoid irradiation, or carmustine. A median number of 4.6 x 10(6) CD34+ cells/kg were obtained with a median of three leukapheresis procedures. Mobilization of PBPCs using chemotherapy plus G-CSF was superior to G-CSF alone (6.7 x 10(6) versus 1.5 x 10(6) CD34+ cells/kg; P = 0.0002). Poorer mobilization of progenitor cells was observed in patients who had previously received stem cell-toxic chemotherapy, including (a) nitrogen mustard, procarbazine, melphalan, carmustine or > 7.5 g of cytarabine chemotherapy premobilization (2.0 x 10(6) versus 6.0 x 10(6) CD34+ cells/kg; P = 0.005), or (b) > or = 11 cycles of any previous chemotherapy (2.6 x 10(6) versus 6.7 x 10(6) CD34+ cells/kg; P = 0.02). Platelet recovery to > 20,000/microliter was delayed in patients who received < 2.0 x 10(6) CD34+ cells (median, 13 versus 22 days; P = 0.06). Patients who received > or = 11 cycles of chemotherapy prior to PBPC mobilization tended to have delayed platelet recovery to > 20,000/microliter and to require more platelet transfusions than less extensively pretreated patients (median, 13.5 versus 23.5 days; P = 0.15; median number of platelet transfusion episodes, 13 versus 9; P = 0.17). These data suggest that current strategies to mobilize PBPCs may be suboptimal in patients who have received either stem cell-toxic chemotherapy or > or = 11 cycles of chemotherapy prior to PBPC mobilization. Alternative approaches, such as ex vivo expansion or the use of other growth factors in addition to G-CSF, may improve mobilization of progenitor cells for PBPC transplantation.


Perceptions and impact of bipolar disorder in Japan: results of an Internet survey.
Bipolar disorder is a recurrent and episodic illness. This survey study assessed experiences and identified clinical insights of individuals with bipolar disorder. An Internet-based monitor system database was screened for patients with bipolar disorder in Japan (February and March 2013). Of 1,050 patients, 457 completed surveys, and results were analyzed with descriptive statistics. Approximately one-fourth of respondents were diagnosed with bipolar disorder on their first visit to medical institutions, although the most common initial diagnosis was depression/depressive state (65%). Mean time lag between first-time visit to a medical institution and receipt of correct diagnosis of bipolar disorder was 4 years; one-third of patients experienced more than 5 years of lag time. Three perceived reasons for lapsed time before correct diagnosis were "(patients) Did not consider manic symptoms as illness, and did not tell the doctor about them," "I (patient) did not know of bipolar disorder," and "Lack of communication between my doctor and myself (patient)." Among participants who believed that they were initially incorrectly diagnosed and improperly treated, most experienced socioeconomic problems, such as having long-term inability to work or to study (65%). Sources of encouragement for participants included "To have someone to consult with" (41%) followed by having "People around me treat me the same as before" (40%). Individuals with bipolar disorder reported a time lag of many years before accurate diagnosis, and substantial burden imposed by the illness. Encouragement should be provided for individuals to live positively with bipolar disorder.


Increased food intake and changes in metabolic hormones in response to chronic sleep restriction alternated with short periods of sleep allowance.
Rodent models for sleep restriction have good face validity when examining food intake and related regulatory metabolic hormones. However, in contrast to epidemiological studies in which sleep restriction is associated with body weight gain, sleep-restricted rats show a decrease in body weight. This difference with the human situation might be caused by the alternation between periods of sleep restriction and sleep allowance that often occur in real life. Therefore, we assessed the metabolic consequences of a chronic sleep restriction protocol that modeled working weeks with restricted sleep time alternated by weekends with sleep allowance. We hypothesized that this protocol could lead to body weight gain. Male Wistar rats were divided into three groups: sleep restriction (SR), forced activity control (FA), and home cage control (HC). SR rats were subjected to chronic sleep restriction by keeping them awake for 20 h per day in slowly rotating drums. To model the human condition, rats were subjected to a 4-wk protocol, with each week consisting of a 5-day period of sleep restriction followed by a 2-day period of sleep allowance. During the first experimental week, SR caused a clear attenuation of growth. In subsequent weeks, two important processes occurred: 1) a remarkable increase in food intake during SR days, 2) an increase in weight gain during the weekends of sleep allowance, even though food intake during those days was comparable to controls. In conclusion, our data revealed that the alternation between periods of sleep restriction and sleep allowance leads to complex changes in food intake and body weight, that prevent the weight loss normally seen in continuous sleep-restricted rats. Therefore, this "week-weekend" protocol may be a better model to study the metabolic consequences of restricted sleep.


Solvent-induced electrochemistry at an electrically asymmetric carbon Janus particle.
Chemical doping through heteroatom substitution is often used to control the Fermi level of semiconductor materials. Doping also occurs when surface adsorbed molecules modify the Fermi level of low dimensional materials such as carbon nanotubes. A gradient in dopant concentration, and hence the chemical potential, across such a material generates usable electrical current. This opens up the possibility of creating asymmetric catalytic particles capable of generating voltage from a surrounding solvent that imposes such a gradient, enabling electrochemical transformations. In this work, we report that symmetry-broken carbon particles comprised of high surface area single-walled carbon nanotube networks can effectively convert exothermic solvent adsorption into usable electrical potential, turning over electrochemical redox processes in situ with no external power supply. The results from ferrocene oxidation and the selective electro-oxidation of alcohols underscore the potential of solvent powered electrocatalytic particles to extend electrochemical transformation to various environments.


[Long-acting injectable antipsychotics: an expert opinion from the Association des médecins psychiatres du Québec].
To present points of agreement and disagreement about antipsychotics. Since the appearance of 2nd generation long-acting antipsychotics (LAA), and given the high frequency of noncompliance with antipsychotics in psychotic disorders, LAAs have attracted more interest in psychiatric literature. However,their use is suboptimal, globally, and is also subject to significant national disparities. ln this context,the Association des médecins psychiatres du Québec (AMPQ) has asked for a review of the evidence concerning LAA efficiency and tolerance, and has called for consensual c1inical reflection on the benefits and obstacles of prescribing them, as weil as potential solutions, including administrative and judiciary dimensions. The AMPQ established an expert committee, from 4 Quebec universities, which was responsible for preparing the review paper. The committee intended to appropriately provide c1inicians with the different aspects of LAA use. The committee produced a qualitative and selective review. Mean LAA prescription rates observed in Canada are around 6% and data to confirm this are scarce. A 15% to 25% rate could be suggested. The committee has submitted the Quebec long-acting antipsychotic algorithm (QAAPAPLE, derived from the French acronym) as a result of the consensus reached by the 4 university psychiatry departments.


Enzymatic capacities of metabolic fuel use in cuttlefish (Sepia officinalis) and responses to food deprivation: insight into the metabolic organization and starvation survival strategy of cephalopods.
Food limitation is a common challenge for animals. Cephalopods are sensitive to starvation because of high metabolic rates and growth rates related to their "live fast, die young" life history. We investigated how enzymatic capacities of key metabolic pathways are modulated during starvation in the common cuttlefish (Sepia officinalis) to gain insight into the metabolic organization of cephalopods and their strategies for coping with food limitation. In particular, lipids have traditionally been considered unimportant fuels in cephalopods, yet, puzzlingly, many species (including cuttlefish) mobilize the lipid stores in their digestive gland during starvation. Using a comprehensive multi-tissue assay of enzymatic capacities for energy metabolism, we show that, during long-term starvation (12 days), glycolytic capacity for glucose use is decreased in cuttlefish tissues, while capacities for use of lipid-based fuels (fatty acids and ketone bodies) and amino acid fuels are retained or increased. Specifically, the capacity to use the ketone body acetoacetate as fuel is widespread across tissues and gill has a previously unrecognized capacity for fatty acid catabolism, albeit at low rates. The capacity for de novo glucose synthesis (gluconeogenesis), important for glucose homeostasis, likely is restricted to the digestive gland, contrary to previous reports of widespread gluconeogenesis among cephalopod tissues. Short-term starvation (3-5 days) had few effects on enzymatic capacities. Similar to vertebrates, lipid-based fuels, putatively mobilized from fat stores in the digestive gland, appear to be important energy sources for cephalopods, especially during starvation when glycolytic capacity is decreased perhaps to conserve available glucose.


Uncovering ecosystem service bundles through social preferences.
Ecosystem service assessments have increasingly been used to support environmental management policies, mainly based on biophysical and economic indicators. However, few studies have coped with the social-cultural dimension of ecosystem services, despite being considered a research priority. We examined how ecosystem service bundles and trade-offs emerge from diverging social preferences toward ecosystem services delivered by various types of ecosystems in Spain. We conducted 3,379 direct face-to-face questionnaires in eight different case study sites from 2007 to 2011. Overall, 90.5% of the sampled population recognized the ecosystem's capacity to deliver services. Formal studies, environmental behavior, and gender variables influenced the probability of people recognizing the ecosystem's capacity to provide services. The ecosystem services most frequently perceived by people were regulating services; of those, air purification held the greatest importance. However, statistical analysis showed that socio-cultural factors and the conservation management strategy of ecosystems (i.e., National Park, Natural Park, or a non-protected area) have an effect on social preferences toward ecosystem services. Ecosystem service trade-offs and bundles were identified by analyzing social preferences through multivariate analysis (redundancy analysis and hierarchical cluster analysis). We found a clear trade-off among provisioning services (and recreational hunting) versus regulating services and almost all cultural services. We identified three ecosystem service bundles associated with the conservation management strategy and the rural-urban gradient. We conclude that socio-cultural preferences toward ecosystem services can serve as a tool to identify relevant services for people, the factors underlying these social preferences, and emerging ecosystem service bundles and trade-offs.


CADASIL-associated Notch3 mutations have differential effects both on ligand binding and ligand-induced Notch3 receptor signaling through RBP-Jk.
Mutations in the NOTCH3 gene are the cause of cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL), a hereditary angiopathy leading to strokes and dementia. Pathogenic mutations remove or insert cysteine residues within epidermal growth factor (EGF) repeats in the extracellular domain of the Notch3 receptor (N3ECD). Vascular smooth muscle cells (VSMC) are the predominant site of Notch3 expression in adults. In CADASIL patients, VSMC degenerate and N3ECD is deposited within the vasculature. However, the mechanisms underlying VSMC degeneration and N3ECD accumulation are still unknown. In this study, we investigated the consequences of three pathogenic Notch3 mutations on the biological activity of the receptor by analyzing ligand (Delta-/Jagged-)-induced signaling via RBP-Jk. Two mutations (R133C and C183R) that are located outside the putative ligand binding domain (LBD) of the receptor were found to result in normal Jagged1-induced signaling in A7r5 VSMC, whereas the third mutation (C455R located within the putative LBD) showed strongly reduced signaling activity. Ligand binding assays with soluble Delta1 and Jagged1 revealed that C455R interferes with ligand binding through disruption of the LBD which, as we show here, is located in EGF repeats 10/11 of Notch3. All mutant receptors including Notch3C455R were targeted to the cell surface but showed an elevated ratio between the unprocessed full-length 280-kDa receptor and S1-cleaved receptor fragments. Taken together, these data indicate that CADASIL-associated Notch3 mutations differ with respect to their consequences both on ligand binding and ligand-induced signaling through RBP-Jk, whereas they have similar effects on receptor maturation. Moreover, the data suggest that ligand-induced receptor shedding may not be required for N3ECD deposition in CADASIL.


Nickel for your thoughts: urey and the origin of the moon.
The theories of Harold C. Urey (1893-1981) on the origin of the moon are discussed in relation to earlier ideas, especially George Howard Darwin's fission hypothesis. Urey's espousal of the idea that the moon had been captured by the earth and has preserved information about the earliest history of the solar system led him to advocate a manned lunar landing. Results from the Apollo missions, in particular the deficiency of siderophile elements in the lunar crust, led him to abandon the capture selenogony and tentatively adopt the fission hypothesis.


Weighted density-functional theory for simple fluids: prewetting of a Lennard-Jones fluid.
The prewetting of a Lennard-Jones fluid is studied using weighted density-functional theory. The intrinsic Helmholtz free-energy functional is separated into repulsive and attractive contributions. An accurate functional for hard spheres is used for the repulsive functional and a weighted density-functional method is used for the attractive part. The results for this theory are compared against mean-field density-functional theory, the theory of Velasco and Tarazona [E. Velasco and P. Tarazona, J. Chem. Phys. 91, 7916 (1989)] and grand canonical ensemble simulation results. The results demonstrate that the weighted density functional for attractive forces may offer a significant increase in accuracy over the other theories. The density-functional and simulation results also indicate that a previous estimate of the wetting temperature for a model of the interaction of argon with solid carbon dioxide, obtained from simulations [J. E. Finn and P. A. Monson, Phys. Rev. A, 39, 6402 (1989)], is incorrect. The weighted density-functional method indicates that triple-point prewetting is observed for this model potential.


State-of-the-art chronic thromboembolic pulmonary hypertension diagnosis and management.
Chronic thromboembolic pulmonary hypertension (CTEPH) is a debilitating disease caused by chronic obstruction of pulmonary artery branches following episodes of pulmonary embolism and incomplete thrombus resolution. The prognosis of patients with CTEPH is poor unless an early diagnosis is made and treatment is initiated. Chest radiography and echocardiography are used in the initial assessment of suspected pulmonary hypertension. A diagnosis of CTEPH may be confirmed by the presence of a mismatched wedge-shaped perfusion deficit during ventilation/perfusion scintigraphy or characteristic findings during multi-slice computed tomography (CT) angiography, including a mosaic perfusion pattern, dilatation of proximal pulmonary arteries and right heart chambers, and the presence of vascular stenosis or obstruction. Prior to possible surgery, pulmonary angiography remains the definitive diagnostic technique, indicating the site and accessibility of the obstruction. However, many centres utilise CT and magnetic resonance imaging following recent advances in these noninvasive techniques. Haemodynamic evaluation via right heart catheterisation is also mandatory, as pulmonary vascular resistance is the most important determinant of both prognosis and the risk associated with pulmonary endarterectomy surgery. Accurate CTEPH diagnosis and characterisation of its extent and distribution are imperative to allow the prompt initiation of treatment, particularly surgical pulmonary endarterectomy in eligible patients.


Staurosporine enhances gonadotrophin-releasing hormone-stimulated luteinizing hormone secretion.
In intact sheep gonadotropes, the protein kinase inhibitor, staurosporine, inhibited the stimulatory effect of phorbol 12-myristate 13-acetate (PMA) on luteinizing hormone (LH) secretion. Under the same conditions staurosporine enhanced gonadotrophin-releasing hormone (GnRH)-stimulated LH exocytosis without altering the EC50 of GnRH and without affecting basal LH exocytosis. These results suggest that PKC does not play a major role in mediating acute GnRH-stimulated LH exocytosis. Furthermore, they demonstrate that staurosporine enhances GnRH stimulus-secretion coupling. Both extracellular Ca2(+)-dependent and Ca2(+)-independent components of GnRH-stimulated LH secretion were enhanced by the drug. Staurosporine had no effect on GnRH stimulation of cAMP and inositol phosphate synthesis. In permeabilized cells staurosporine did not enhance Ca2(+)- and cAMP-stimulated LH exocytosis. Based on these results we hypothesize that staurosporine inhibits a protein kinase which is activated by GnRH and which negatively modulates GnRH stimulus-secretion coupling.


A novel synchronous fluorescence spectroscopic approach for the rapid determination of three polycyclic aromatic hydrocarbons in tea with simple microwave-assisted pretreatment of sample.
Many polycyclic aromatic hydrocarbons (PAHs) are carcinogenic, and some have been reported to be present in tea. People can be exposed to PAHs through tea consumption. Therefore, there is real importance for the determination of PAHs in tea. Because of the complex matrix of tea, it is hard to detect PAHs in tea without cleanup and chromatographic separation procedures. In this research, for the first time, a novel synchronous fluorescence spectroscopic approach coupling nonlinear variable-angle synchronous and matrix-isopotential synchronous scanning modes has been developed for the rapid determination of benzo(a)pyrene (BaP), benzo(k)fluoranthene (BkF), and anthracene (AN) in tea with simple microwave-assisted pretreatment of samples. This novel technique is able to resolve the spectra of the three PAHs well, even with interference from other EPA PAHs. The detection limits for BaP, BkF, and AN in tea were 0.18-0.28, 0.55-0.89, and 0.64-3.58 μg/kg, respectively, depending on various teas, with satisfactory recoveries ranging from 77.1 to 116%. The relative standard deviations achieved for BaP, BkF, and AN were 1.5, 6.6, and 8.5% for green tea; 2.9, 7.4, and 2.1% for oolong tea; and 5.6, 5.4, and 5.8% for black tea, respectively. Our results showed good correlation with those of gas chromatography-mass spectrometry. The approach developed is simple, reliable, and cost-efficient, providing an attractive alternative for the rapid selective screening of PAHs in tea.


Anatomy and development of the koala, Phascolarctos cinereus: an evolutionary perspective on the superfamily Vombatoidea.
Fifteen koalas (Phascolarctos cinereus)--5 pouched young from 4 to 6.5 months and 10 adults from 5 to 16.5 years--were analyzed for functional parameters (body composition, limb segment and muscle mass, post-cranial skeletal characters) and developmental expressions (growth of body, brain, musculature). These data were compared with a convergent eutherian, the three-toed sloth, Bradypus infuscatus, and with the koala's distant (Macropodid; wallabies) and proximate (Vombatid; wombats) marsupial relatives. Musculoskeletal structures correlated with sitting and climbing; the growth of the young and the physiological demands of adulthood correlated with the low-quality diet of Eucalyptus foliage. The gestalt of the ancestral Vombatoids (pronograde quadrupeds, generalist browsers and social conservatives with low basal metabolism and attenuated development) provided the baseline essential for their locomotor and nutritional divergence into arboreal browsers, the koalas, and fossorial grazers, the wombats.


Genetics and public health: the experience of a reference center for diagnosis of 22q11.2 deletion in Brazil and suggestions for implementing genetic testing.
Considering the prevalence of 22q11.2 deletion syndrome (22q11.2 DS) of around 1:4,000 and of palatal abnormalities in 70 % of the cases of 22q11.2 DS and taking into account the Brazilian health system and its current situation of medical genetic services, this study aims to contribute to establish strategies for genetic diagnosis. The access to genetic testing at 11 services was investigated and samples from 100 patients with palatal abnormalities and suspicion of 22q11.2 DS were sent to a reference center. Laboratorial techniques included karyotyping, fluorescence in situ hybridization (FISH), and multiplex ligation-dependent probe amplification. Costs were also calculated. Disparities among centers for genetic diagnosis were evident, with remarkable regional differences. Some of the obstacles encountered were difficulties for families to show up for medical appointments, complementary evaluations, and for the clinics to send the samples to the reference center. A conclusive diagnosis was reached for 38 % of patients. Combination of karyotyping and FISH had better laboratorial cost-effectiveness. These results might represent the reality for the investigation of other genetic conditions. Clinical and laboratorial approaches herein presented could be adapted for use under different genetic conditions in the Brazilian health system, which has relatively limited financial and human resources. Suggestions for the rational implementation of genetic testing in developing countries are presented.


Children and Young People's Health Partnership (CYPHP) Evelina London model of care: protocol for an opportunistic cluster randomised controlled trial (cRCT) to assess child health outcomes, healthcare quality and health service use.
Children and young people (CYP) in many high-income settings have poor healthcare outcomes, especially those with long-term conditions (LTCs). Emergency and outpatient hospital service use is increasing unsustainably. To address these problems, the Children and Young People's Health Partnership (CYPHP) has developed and is evaluating an integrated model of care as part of a health systems strengthening programme across two boroughs of London, UK that are characterised by mixed ethnic populations and varying levels of deprivation. The CYPHP Evelina London model of care comprises proactive case-finding and triage, specialist clinics and transformative education and training for professionals working with CYP. Services are delivered by multidisciplinary health teams with an emphasis on increased coordination across primary, community and hospital settings and integration of physical and mental healthcare that accounts for the CYP's social context. The phased roll out of the CYPHP Evelina London model allows an opportunistic population-based evaluation using a cluster randomised controlled trial design. Seventy general practices across two London boroughs, grouped into 23 clusters, were randomised to provide either the CYPHP model of care (n=11) or enhanced usual care (n=12).The evaluation will measure the impact of the CYPHP Evelina London model of care on child and parent health and well-being, healthcare quality and health service use up to 2 years postimplementation. A population-level evaluation will use routinely collected pseudonymised healthcare data to conduct a service-use analysis for all CYP registered with a participating general practice (n=~90 000) with the rate of non-elective admissions as the primary outcome. We will seek consent from a subset of this population, with specific conditions (target n=2138) to assess the impact on patient-reported outcomes using the Paediatric Quality of Life Inventory (PedsQL) and Warwick-Edinburgh Mental Well-Being Scale (WEBWMS) as, respectively, the child- and parent-related primary outcomes. Ethics approval obtained from South West-Cornwall & Plymouth Research Ethics Committee. Results will be submitted for publication in peer-reviewed journals. Findings will be generalisable to community-based models of care, especially in urban settings. Our process evaluation will identify barriers and enablers of implementation and delivery of care salient to the context and condition. NCT03461848; Pre-results.


Fast and continuous plasma extraction from whole human blood based on expanding cell-free layer devices.
This paper presents promising microfluidic devices designed for continuous and passive extraction of plasma from whole human blood. These designs are based on red cells lateral migration and the resulting cell-free layer locally expanded by geometric singularities such as an enlargement of the channel or a cavity adjacent to the channel. After an explanation of flow patterns, different tests are described that confirm the advantages of both proposed singularities, providing a 1.5 and 2X increase in extraction yield compared to a reference device, for 1:20 diluted blood at 100 microL/min. Devices have also been successively optimized, with extraction yields up to 17.8%, and biologically validated for plasma extraction, with no protein loss or denaturation, no hemolysis and with excellent cell purity. Finally, the dilution effect has been experimentally investigated.


Comparison of sun pillars with light pillars from nearby light sources.
Using a Monte Carlo method, we simulate the appearance of light pillars produced by nearby light sources and compare their appearance with simulations of Sun pillars. Photographs of light pillars are also compared with the simulations. We expand the idea of light and Sun pillars by examining the reflected-light patterns from several different known distributions of airborne ice crystals. Polarization properties of light pillars from nearly horizontally oriented plate crystals are also simulated.


Homogeneous degradation of 1,2,9,10-tetrachlorodecane in aqueous solutions using hydrogen peroxide, iron and UV light.
The homogeneous degradation of the polychlorinated n-alkane, 1,2,9,10-tetrachlorodecane (T4C10), was studied in aqueous solutions of hydrogen peroxide, including Fenton and photo-Fenton reaction conditions. All solutions were adjusted to a pH of 2.8 and an ionic strength of 0.1 M NaClO4 prior to photolysis. T4C10 (2 x 10(-6) M) was substantially degraded by the H2O2/UV system (1.0 x 10(-2) M H2O2), with 60% disappearance in 20 min of irradiation in a photoreactor equipped with 300 nm lamps of light intensity 3.6 x 10(-5) Ein L(-1) min(-1) (established by ferrioxalate actinometry). The reaction produced stoichiometric amounts of chloride ion indicating complete dechlorination of the chlorinated n-alkane. T4C10 degraded very slowly under Fenton (Fe2+/H2O2/dark) and Fenton-like (Fe3+/H2O2/dark) conditions. However, when the same solutions were irradiated, T4C10 degraded more rapidly than in the H2O2/UV system, with 61% disappearance in 10 min of exposure. The rapid degradation is related to the enhanced degradation of hydrogen peroxide to oxidizing *OH radicals under photo-Fenton conditions. Degradation was inhibited in both the H2O2/UV and photo-Fenton systems by the addition of KI and tert-butyl alcohol due to *OH scavenging.


Evidence for a repair enzyme complex involving ERCC1 and complementing activities of ERCC4, ERCC11 and xeroderma pigmentosum group F.
Nucleotide excision repair (NER), one of the major cellular DNA repair systems, removes a wide range of lesions in a multi-enzyme reaction. In man, a NER defect due to a mutation in one of at least 11 distinct genes, can give rise to the inherited repair disorders xeroderma pigmentosum (XP), Cockayne's syndrome or PIBIDS, a photosensitive form of the brittle hair disease trichothiodystrophy. Laboratory-induced NER-deficient mutants of cultured rodent cells have been classified into 11 complementation groups (CGs). Some of these have been shown to correspond with human disorders. In cell-free extracts prepared from rodent CGs 1-5 and 11, but not in a mutant from CG6, we find an impaired repair of damage induced in plasmids by UV light and N-acetoxy-acetylaminofluorene. Complementation analysis in vitro of rodent CGs is accomplished by pairwise mixing of mutant extracts. The results show that mutants from groups 2, 3, 5 and XP-A can complement all other CGs tested. However, selective non-complementation in vitro was observed in mutual mixtures of groups 1, 4, 11 and XP-F, suggesting that the complementing activities involved somehow affect each other. Depletion of wild-type human extracts from ERCC1 protein using specific anti-ERCC1 antibodies concomitantly removed the correcting activities for groups 4, 11 and XP-F, but not those for the other CGs. Furthermore, we find that 33 kDa ERCC1 protein sediments as a high mol. wt species of approximately 120 kDa in a native glycerol gradient.(ABSTRACT TRUNCATED AT 250 WORDS)


Interaction of Caffeic Acid with SDS Micellar Aggregates.
Micellar systems consisting of a surfactant and an additive such as an organic salt or an acid usually self-organize as a series of worm-like micelles that ultimately form a micellar network. The nature of the additive influences micellar structure and properties such as aggregate lifetime. For ionic surfactants such as sodium dodecyl sulfate (SDS), CMC decreases with increasing temperature to a minimum in the low-temperature region beyond which it exhibits the opposite trend. The presence of additives in a surfactant micellar system also modifies monomer interactions in aggregates, thereby altering CMC and conductance. Because the standard deviation of β was always lower than 10%, its slight decrease with increasing temperature was not significant. However, the absolute value of Gibbs free enthalpy, a thermodynamic potential that can be used to calculate the maximum of reversible work, increased with increasing temperature and caffeic acid concentration. Micellization in the presence of caffeic acid was an endothermic process, which was entropically controlled. The enthalpy and enthropy positive values resulted from melting of "icebergs" or "flickering clusters" around the surfactant, leading to increased packing of hydrocarbon chains within the micellar core in a non-random manner. This can be possibly explained by caffeic acid governing the 3D matrix structure of water around the micellar aggregates. The fact that both enthalpy and entropy were positive testifies to the importance of hydrophobic interactions as a major driving force for micellization. Micellar systems allow the service life of some products to be extended without the need to increase the amounts of post-harvest storage preservatives used. If a surfactant is not an allowed ingredient or food additive, carefully washing it off before the product is consumed can avoid any associated risks. In this work, we examined the influence of temperature and SDS concentration on the properties of SDS⁻caffeic acid micellar systems. Micellar properties can be modified with various additives to develop new uses for micelles. This allows smaller amounts of additives to be used without detracting from their benefits.


Miniaturization: an overview of biotechnologies for monitoring the physiology and pathophysiology of rodent animal models.
Recent advances in bioengineering technologies have made it possible to collect high-quality reproducible data quantitatively in a wide range of laboratory animal species, including rodents. Several of these technologies are incorporated into a plan called Miniaturization, which aims to design, develop, and maintain rodent animal models to study the pathophysiology and therapy of human diseases. Laser Doppler flowmetry, digital sonomicrometry, bioelectrical impedance, and microdialysis are some of the most widely used methods under the plan because they cause minimal pain and distress, reduce the number of animals used in biomedical research, and allow chronic, nonterminal assessment of physiological parameters in rodents. An overview of each of these technologies and their major applications in rodents used for biomedical research is provided.


Preparation and application of a poly (ionic liquid)-based molecularly imprinted polymer for multiple monolithic fiber solid-phase microextraction of phenolic acids in fruit juice and beer samples.
In this work, a new molecularly imprinted polymer (MIP) using a poly (ionic liquid) (PIL) as a functional monomer was prepared and utilized as the extraction medium of multiple monolithic fiber solid-phase microextraction (MMF-SPME). A PIL, 1-ally-3-vinylimidazolium chloride, was used as a functional monomer, and 3,4-dihydroxybenzenepropanoic acid (DBA) and ethylene dimethacrylate were chosen as the template molecule and cross-linker, respectively, to synthesize the PIL/MIP. Under the optimized preparation conditions, the recognition coefficient of PIL-MIP for DBA was as high as 11.6. The combination of PIL/MIP-MMF-SPME and high-performance liquid chromatography with diode array detection (HPLC/DAD) was developed for the sensitive determination of phenolic acids (PAs) in fruit juice and beer samples. Under the optimal conditions which were investigated in detail, the limits of detection (LODs, S/N = 3) for PAs in fruit juice were 0.024-0.24 μg L-1, and the related values were 0.011-0.052 μg L-1 for the beer sample. The intra-day and inter-day precision (relative standard deviations, n = 4, %) at 1.0 and 100.0 μg L-1 spiking concentration were both less than 10%. In the analysis of PAs in fruit juice and beer samples, satisfactory recoveries and repeatability were achieved. In comparison with reported approaches, the proposed method exhibited some advantages including high selectivity, convenience, satisfactory sensitivity and environmental friendliness.


The Relationship Between Bruch's Membrane Opening-Minimum Rim Width and Retinal Nerve Fiber Layer Thickness and a New Index Using a Neural Network.
We evaluate the relationship between Bruch's membrane opening minimum rim width (BMO-MRW) and peripapillary retinal nerve fiber layer thickness (pRNFLT) and develop a new parameter combining BMO-MRW and pRNFLT using a neural network to maximize their compensatory values. A total of 402 subjects were divided into two groups: 273 (validation group) and 129 (neural net training) subjects. Linear quadratic and broken-stick regression models were used to explore the relationship between BMO-MRW and pRNFLT. A multilayer neural network was used to create a combined parameter, and diagnostic performances were compared using area under the receiver operating characteristic curves (AUROCs). Regression analyses between BMO-MRW and pRNFLT revealed that the broken-stick model afforded the best fit. Globally, the tipping point was a BMO-MRW of 226.5 μm. BMO-MRW and pRNFLT were correlated significantly with visual field. When differentiating normal from glaucoma subjects, the neural network exhibited the largest AUROC. When differentiating normal from early glaucoma subjects, the overall diagnostic performance decreased, but the neural network still exhibited the largest AUROC. The optimal relationship between BMO-MRW and pRNFLT was revealed using the broken-stick model. Considerable BMO-MRW thinning preceded pRNFLT thinning. The neural network significantly improved diagnostic power by combining BMO-MRW and pRNFLT. A combined index featuring BMO-MRW and pRNFLT data can aid clinical decision-making, particularly when individual parameters yield confusing results. Our neural network effectively combines information from separate parameters.


Intracerebroventricular injection of adeno-associated virus 6 and 9 vectors for cell type-specific transgene expression in the spinal cord.
In the context of motoneuron diseases, gene delivery as an experimental or therapeutic approach is hindered by the challenge to specifically target cell populations that are widely distributed along the spinal cord. Further complicating the task, transgenes often need to be delivered to motoneurons and/or glial cells to address the non-cell-autonomous mechanisms involved in disease pathogenesis. Intracerebroventricular (ICV) injection of recombinant adeno-associated viruses (AAVs) in newborn mice allows distributing viral vectors throughout the central nervous system while limiting undesired transduction of peripheral organs. Here, we show that by combining the appropriate set of AAV serotype and promoter, specific transgene expression can be achieved in either motoneurons or astrocytes along the whole mouse spinal cord. ICV injection of recombinant AAV6 with the cytomegalovirus (cmv) promoter preferentially targets motoneurons, whereas AAV9 particles combined with the astrocyte-specific gfaABC₁D promoter lead to significant transgene expression selectively targeted to astrocytes. Importantly, ICV coinjection of both AAV6-cmv and AAV9-gfaABC₁D results in segregated expression of two different transgenes in motoneurons and astrocytes, respectively. Relevance of viral vector delivery via the cerebrospinal fluid was further investigated in young nonhuman primates. Intracisternal injection of recombinant AAV6-cmv led to robust cervical transduction of motoneurons, highlighting the potential of this approach for gene therapy and modeling of motoneuron diseases.


Isolation and quantification of Quillaja saponaria Molina saponins and lipids in iscom-matrix and iscoms.
In the iscom, multiple copies of antigen are attached by hydrophobic interaction to a matrix which is built up by Quillaja triterpenoid saponins and lipids. Thus, the iscom presents antigen in multimeric form in a small particle with a built-in adjuvant resulting in a highly immunogenic antigen formulation. We have designed a chloroform-methanol-water extraction procedure to isolate the triterpenoid saponins and lipids incorporated into iscom-matrix and iscoms. The triterpenoids in the triterpenoid phase were quantitated using orcinol sulfuric acid detecting their carbohydrate chains and by HPLC. The cholesterol and phosphatidylcholine in the lipid phase were quantitated by HPLC and a commercial colorimetric method for the cholesterol. The quantitative methods showed an almost total separation and recovery of triterpenoids and lipids in their respective phases, while protein was detected in all phases after extraction. The protein content was determined by the method of Lowry and by amino acid analysis. Amino acid analysis was shown to be the reliable method of the two to quantitate proteins in iscoms. In conclusion, simple, reproducible and efficient procedures have been designed to isolate and quantitate the triterpenoids and lipids added for preparation of iscom-matrix and iscoms. The procedures described should also be useful to adequately define constituents in prospective vaccines.


Mechanisms of Chinese herb emodin and somatostatin analogs on pancreatic regeneration in acute pancreatitis in rats.
Studies have revealed that emodin is a potent agent in the management of clinical and experimental acute pancreatitis, but the molecular mechanisms by which emodin produces its biologic effects, especially on pancreatic regeneration after acute pancreatitis, remain unknown. Numerous experimental and clinical studies have shown that somatostatin analogs have favorable effects on acute pancreatitis, but their role in the management of acute pancreatitis remains controversial. To investigate mechanisms of the Chinese herb emodin and somatostatin analogs (SSa; Sandostatin) in acute pancreatitis of rats by analyzing the changes in pancreatic tissue cytokine transforming growth factor beta1 (TGFbeta1) and epidermal growth factor (EGF) gene expression, DNA synthesis, total protein content, and the relations between them. Acute pancreatitis was induced by intraperitoneal infusion of cerulein in rats. Emodin was administered intravenously and Sandostatin was administered subcutaneously at the time of induction of pancreatitis and 24, 48, and 72 hours afterward. Rats were killed at 6, 24, 48, 72, and 96 hours after the operation. The mRNA expression of TGFbeta1 and EGF were evaluated by reverse transcription polymerase chain reaction, and pancreatic tissue DNA synthesis was measured by the 3H-thymidine incorporation method in vitro. Total protein content was detected by Lowry's method. The serum amylase level was decreased significantly in the emodin-treated and Sandostatin-treated groups in comparison with the nontreated group. Pancreatic tissue DNA synthesis was significantly decreased at 72 hours after the induction of pancreatitis, and a marked increase was observed at 96 hours after treatment with emodin and Sandostatin. Within 48 hours of the induction of pancreatitis, the total protein content in pancreatic tissue declined, but there was a remarkable increase in the emodin-treated group at 96 hours and Sandostatin-treated group at 48 hours. Expression of TGFbeta1 mRNA and EGF mRNA were undetectable in normal pancreas and the nontreated group at 6 hours but was observed from 24 hours to 96 hours after the induction of pancreatitis and reached its maximum at 72 hours. TGFbeta1 mRNA could be detected 6 hours after treatment with emodin and Sandostatin, and its expression was significantly higher in the emodin-treated and Sandostatin-treated groups than in the nontreated group at 24 and 48 hours. The expression of EGF mRNA was significantly higher in the emodin-treated and Sandostatin-treated group than in the nontreated group at 48 hours. It was concluded that mechanisms of the Chinese herb emodin and somatostatin analogs in the management of acute pancreatitis in rats might be ascribed to the upregulation of TGFbeta1 and EGF gene expression, which subsequently increases DNA synthesis and protein content and thus accelerates pancreatic repair and regeneration.


Putative perception of rotating permanent magnetic fields following ingestion of LSD.
While sitting alone in complete darkness, 3 participants who had ingested psychotropic concentrations of lysergic acid diethylamide reported diffuse blobs of white, purplish, or greenish-yellow lights as two horseshoe magnets rotated at 0.5 Hz. The experiences were not reported when the magnets were stationary or removed from the apparatus. The estimated peak-to-peak variation in field strength at the distance of perception was between 50 and 500 nanoTesla. An association between these results and possible ergot-induced perceptions of "magnet light" reported during the last century by von Reichenbach (1851) is suggested.


kappa-opioid receptor agonists modulate excitatory transmission in substantia gelatinosa neurons of the rat spinal cord.
This study examined the effects of selective activation of kappa 1-opioid receptors on excitatory transmission in substantia gelatinosa (SG) using intracellular recordings from SG neurons in transverse slices of the young rat lumbar spinal cord. Monosynaptic and polysynaptic excitatory postsynaptic potentials (EPSPs) were evoked by orthodromic electrical stimulation of A delta or C primary afferent fibers in the dorsal root after blocking inhibitory inputs with bicuculline and strychnine, NMDA receptors with D-2-amino-5-phosphonovaleric acid and mu- and delta-opioid receptors with CTAP and ICI 174,864, respectively. Bath application of dynorphin A1-17 or U-69, 593 caused dual modulation of the peak amplitude of presumed monosynaptic AMPA receptor-mediated EPSPs, decreasing synaptic potentials at nanomolar concentrations in a majority of SG cells examined (dynorphin, 63%; U-69,593, 91%), and increasing EPSPs at micromolar concentrations. Only the inhibitory action of dynorphin A1-17 was consistently and completely blocked by norbinaltorphimine (nor-BNI). Since U-69,593 and nor-BNI are selective for the kappa 1-opioid receptors, the depression of EPSPs is likely to be mediated by the kappa1-opioid receptors. Under conditions of blockade of synaptic transmission with TTX and mu-and delta-opioid receptors, dynorphin A1-17 and U-69,593 hyperpolarize most of SG neurons and decrease their membrane input resistance, the finding suggesting that direct interaction of kappa-agonists with a postsynaptic receptor is likely explanation for the inhibition of EPSPs. However, in some SG cells, the inhibition of EPSPs appears to be of presynaptic origin since dynorphin A1-17 and U-69,593 did depress the EPSPs in the absence of changes in passive membrane properties. Rp-cAMPS, a membrane permeant potent competitive inhibitor of cAMP-activated protein kinase, prevented the depressant effect of dynorphin A 1-17. This finding suggested a possibility that dynorphin A1-17, acting through a decrease in intracellular cyclic AMP levels, can reduce the synaptic responses of SG neurons. These results provide the first electrophysiological demonstration that the activation of kappa 1-opioid receptors inhibits AMPA receptor-mediated primary afferent neurotransmission in the substantia gelatinosa of the young rat spinal cord. This effect may mediate the ability of kappa-receptor agonists to produce antinociception.


Influence of surgery on deglutitive upper oesophageal sphincter mechanics in Zenker's diverticulum.
To evaluate the role of upper oesophageal sphincter (UOS) compliance in dysphagia, the functional consequences of surgery were evaluated in eight patients with pharyngeal diverticula. The study examined the hypotheses that hypopharyngeal intrabolus pressure is an indicator of UOS compliance and that UOS opening and intrabolus pressure are normalised by surgery. In eight patients and nine healthy controls, we measured the timing of swallow events, UOS relaxation, maximal UOS dimensions, intrabolus pressure, and trans-sphincteric bolus flow rates by simultaneous videoradiography and pharyngeal manometry. Bolus flow rates were not changed by surgery. Surgery significantly increased UOS opening (p = 0.0001) and reduced hypopharyngeal intrabolus pressure (p = 0.0001). The slope of the relation between sphincter area and intrabolus pressure was steeper in patients than controls and was normalised by surgery. Surgery had minor effects on basal UOS tone and timing of swallow events. Upper oesophageal sphincter compliance is poor in Zenker's diverticulum and is normalised by surgery. Hypopharyngeal intrabolus pressure, which correlates with resistance to trans-sphincteric bolus flow, is a useful indicator of UOS compliance. Intrabolus pressure may be a predictor of outcome after myotomy in pharyngeal dysphagia. Cricopharyngeal myotomy is a mandatory component of surgery for Zenker's diverticulum.


Early manifestation of calcinosis cutis in pseudohypoparathyroidism type Ia associated with a novel mutation in the GNAS gene.
To clarify the molecular defect for the clinical finding of congenital hypothyroidism combined with the manifestation of calcinosis cutis in infancy. The male patient presented with moderately elevated blood thyrotropin levels at neonatal screening combined with slightly decreased plasma thyroxine and tri-iodothyronine concentrations, necessitating thyroid hormone substitution 2 weeks after birth. At the age of 7 months calcinosis cutis was seen and the patient underwent further investigation. Typical features of Albright's hereditary osteodystrophy (AHO), including round face, obesity and delayed psychomotor development, were found. Laboratory investigation revealed a resistance to parathyroid hormone (PTH) with highly elevated PTH levels and a reduction in adenylyl cyclase-stimulating protein (Gsalpha) activity leading to the diagnosis of pseudohypoparathyroidism type Ia (PHP Ia). A novel heterozygous mutation (c364T > G in exon 5, leading to the amino acid substitution Ile-106 --> Ser) was detected in the GNAS gene of the patient. This mutation was not found in the patient's parents, both of whom showed normal Gsalpha protein activity in erythrocytes and no features of AHO. A de novo mutation is therefore likely. Subcutaneous calcifications in infancy should prompt the clinician to a thorough search for an underlying disease. The possibility of AHO and PHP Ia should be considered in children with hypothyroidism and calcinosis cutis. Systematic reviews regarding the frequency of calcinosis in AHO are warranted.


Colonoscopy in the very elderly: a review of 157 cases.
Colonoscopy is currently the best diagnostic modality for evaluating colonic diseases but studies of its use in the very elderly are limited. A single-institution review of all patients aged 85 years or older who underwent colonoscopy from June 2003 to June 2005 was performed. Parameters evaluated included indications for colonoscopy, findings, ability to perform a complete colonoscopy, and immediate and delayed (< or =21 days) complications. A total of 157 patients aged 85 years or older (median = 87, range = 85-99) underwent colonoscopy during the two-year period. The cecal intubation rate was 90%. Number of cancers detected/indications for colonoscopy include gross or occult bleeding per rectum, 3/51 (5.9%); abnormal physical exam, 1/2 (50%); abnormal abdominal computed tomography, 3/5 (60%); anemia, 1/25 (4.0%); screening, 0/14; previous history of colonic malignancy, 0/10; previous history of polyps, 0/21; change in bowel habits, 0/5; family history of colonic malignancy, 0/6; abdominal pain, 0/4; diarrhea, 0/6; fecal impaction, 0/2; unknown, 0/6. Immediate complications included hemorrhage at a polypectomy site in one patient that was controlled endoscopically, one episode of bradycardia, and one incident of atrial fibrillation. There were no delayed complications resulting from colonoscopy. Our data suggest that colonoscopy can be safely and successfully performed in the very elderly. In patients with symptoms or suggestive radiographic findings, cancer was detected in 4.0%-60% of cases. No cases of cancer were discovered in those patients who were asymptomatic.


Advanced imaging among health maintenance organization enrollees with cancer.
Fee-for-service (FFS) Medicare expenditures for advanced imaging studies (defined as computed tomography [CT], magnetic resonance imaging [MRI], positron emission tomography [PET] scans, and nuclear medicine studies [NM]) rapidly increased in the past two decades for patients with cancer. Imaging rates are unknown for patients with cancer, whether under or over age 65 years, in health maintenance organizations (HMOs), where incentives may differ. Incident cases of breast, colorectal, lung, prostate, leukemia, and non-Hodgkin lymphoma (NHL) cancers diagnosed in 2003 and 2006 from four HMOs in the Cancer Research Network were used to determine 2-year overall mean imaging counts and average total imaging costs per HMO enrollee by cancer type for those under and over age 65. There were 44,446 incident cancer patient cases, with a median age of 75 (interquartile range, 71-81), and 454,029 imaging procedures were performed. The mean number of images per patient increased from 7.4 in 2003 to 12.9 in 2006. Rates of imaging were similar across age groups, with the exception of greater use of echocardiograms and NM studies in younger patients with breast cancer and greater use of PET among younger patients with lung cancer. Advanced imaging accounted for approximately 41% of all imaging, or approximately 85% of the $8.7 million in imaging expenditures. Costs were nearly $2,000 per HMO enrollee; costs for younger patients with NHL, leukemia, and lung cancer were nearly $1,000 more in 2003. Rates of advanced imaging appear comparable among FFS and HMO participants of any age with these six cancers.


Peak Ventilation Reference Standards from Exercise Testing: From the FRIEND Registry.
Cardiopulmonary exercise testing (CPX) provides valuable clinical information, including peak ventilation (V˙Epeak), which has been shown to have diagnostic and prognostic value in the assessment of patients with underlying pulmonary disease. This report provides reference standards for V˙Epeak derived from CPX on treadmills in apparently healthy individuals. Nine laboratories in the United States experienced in CPX administration with established quality control procedures contributed to the Fitness Registry and the Importance of Exercise National Database from 2014 to 2017. Data from 5232 maximal exercise tests from men and women without cardiovascular or pulmonary disease were used to create percentiles of V˙Epeak for both men and women by decade between 20 and 79 yr. Additionally, prediction equations were developed for V˙Epeak using descriptive information. V˙Epeak was found to be significantly different between men and women and across age groups (P < 0.05). The rate of decline in V˙Epeak was 8.0% per decade for both men and women. A stepwise regression model of 70% of the sample revealed that sex, age, and height were significant predictors of V˙Epeak. The equation was cross-validated with data from the remaining 30% of the sample with a final equation developed from the full sample (r = 0.73). Additionally, a linear regression model revealed forced expiratory volume in 1 s significantly predicted V˙Epeak (r = 0.73). Reference standards were developed for V˙Epeak for the United States population. Cardiopulmonary exercise testing laboratories will be able to provide interpretation of V˙Epeak from these age and sex-specific percentile reference values or alternatively can use these nonexercise prediction equations incorporating sex, age, and height or with a single predictor of forced expiratory volume in 1 s.


Quantitative tRNA-sequencing uncovers metazoan tissue-specific tRNA regulation.
Transfer RNAs (tRNA) are quintessential in deciphering the genetic code; disseminating nucleic acid triplets into correct amino acid identity. While this decoding function is clear, an emerging theme is that tRNA abundance and functionality can powerfully impact protein production rate, folding, activity, and messenger RNA stability. Importantly, however, the expression pattern of tRNAs is obliquely known. Here we present Quantitative Mature tRNA sequencing (QuantM-tRNA seq), a technique to monitor tRNA abundance and sequence variants secondary to RNA modifications. With QuantM-tRNA seq, we assess the tRNA transcriptome in mammalian tissues. We observe dramatic distinctions in isodecoder expression and known tRNA modifications between tissues. Remarkably, despite dramatic changes in tRNA isodecoder gene expression, the overall anticodon pool of each tRNA family is similar across tissues. These findings suggest that while anticodon pools appear to be buffered via an unknown mechanism, underlying transcriptomic and epitranscriptomic differences suggest a more complex tRNA regulatory landscape.


Serum adiponectin and type 2 diabetes: a 6-year follow-up cohort study.
Studies on factors which may predict the risk of diabetes are scarce. This prospective cohort study was conducted to determine the association between adiponectin and type 2 diabetes among Korean men and women. A total of 42,845 participants who visited one of seven health examination centers located in Seoul and Gyeonggi province, Republic of Korea between 2004 and 2008 were included in this study. The incidence rates of diabetes were determined through December 2011. To evaluate the effects of adiponectin on type 2 diabetes, the Cox proportional hazard model was used. Of the 40,005 participants, 959 developed type 2 diabetes during a 6-year follow-up. After the adjustment for age, body mass index (BMI), and waist circumference, the risks for type 2 diabetes in participants with normoglycemia had a 1.70-fold (95% confidence interval [CI], 1.21 to 2.38) increase in men and a 1.83-fold (95% CI, 1.17 to 2.86) increase in women with the lowest tertile of adiponectin when compared to the highest tertile of adiponectin. For participants with impaired fasting glucose (IFG), the risk for type 2 diabetes had a 1.46-fold (95% CI, 1.17 to 1.83) increase in men and a 2.52-fold (95% CI, 1.57 to 4.06) increase in women with the lowest tertile of adiponectin. Except for female participants with normoglycemia, all the risks remained significant after the adjustment for fasting glucose and other confounding variables. Surprisingly, BMI and waist circumference were not predictors of type 2 diabetes in men or women with IFG after adjustment for fasting glucose and other confounders. A strong association between adiponectin and diabetes was observed. The use of adiponectin as a predictor of type 2 diabetes is considered to be useful.


Metformin decreases high-fat diet-induced renal injury by regulating the expression of adipokines and the renal AMP-activated protein kinase/acetyl-CoA carboxylase pathway in mice.
Metabolic syndrome is characterized by insulin resistance, dyslipidemia and hypertension. These metabolic changes contribute to the development of obesity-induced kidney injury. AMP-activated protein kinase (AMPK) is a ubiquitous enzyme that is involved in the cellular metabolic response to metabolic stress. Metformin, an AMPK activator, has been reported to exert a protective effect against non-alcoholic steatohepatitis. However, little is known about its role in the pathogenesis of obesity-induced renal injury. The aim of this study was to investigate the effects of metformin on high-fat diet (HFD)-induced kidney injury. Obesity was induced by HFD (60% of total calories from fat, 20% protein and 20% carbohydrates) in 6-week-old C57BL/6 mice. Mice were fed HFD plus 0.5% metformin. The effects of metformin on HFD-induced renal injury were evaluated by determining metabolic parameters, serum adipokine levels and renal AMPK/acetyl-CoA carboxylase (ACC) activities, as well as a histological examination. HFD induced metabolic derangement, systemic insulin resistance and glomerular mesangial matrix expansion. The administration of metformin reduced HFD-induced metabolic derangement and renal injury. The administration of metformin reduced the HFD-induced increase in adipokine expression and macrophage infiltration. Moreover, renal AMPK activity, which was decreased by HFD, was recovered following the administration of metformin; in addition, fatty acid oxidation was increased by the inhibition of ACC. These results indicate that metformin exerts beneficial effects on obesity-induced renal injury by regulating systemic inflammation, insulin resistance and the renal AMPK/ACC pathway. The clinical application of metformin to obese or early diabetic patients may be helpful in preventing obesity- or diabetes-related kidney disease.


Effect of Differences in the Primary Structure of the A-Chain on the Aggregation of Insulin Fragments.
Bovine and human insulin have similar primary structures. In this article, the region of the insulin A-chain of bovine and human insulin where the amino acid composition is different was studied. Bovine insulin fragment (BIF) and human insulin fragment (HIF) were synthesized in solid-phase peptide synthesis. The effects of pH, temperature, urea, ionic strength, and stirring on the formation of fibrils were studied using a fractional factorial resolution III experimental design. Fibrillation was monitored by fluorescence and infrared spectroscopy and optical microscopy. Both fragments formed fibrils at pH 1.6 and a temperature of 60 °C. The lag time and apparent aggregation growth rate constant were determined using a two-parameter kinetic model. It was found that the bovine insulin fragment has a shorter lag time than the human insulin one, whereas the exponential phase rate was faster for HIF than for BIF. An increase in β-sheets content with time was observed in both fragments. The increase in β-sheets was preceded by an initial decrease in α-helices followed by an intermediate increase during the transition from the lag phase to elongation phase. Temperature and ionic strength are among the most important experimental factors during the lag phase, whereas ionic strength is replaced by pH during the elongation phase for both the fragments. Congo red binding confirmed the presence of ringlike oligomer structures rich in antiparallel β-sheets, which tend to form fibrils rich in parallel β-sheets.


Emotion processing and the amygdala: from a 'low road' to 'many roads' of evaluating biological significance.
A subcortical pathway through the superior colliculus and pulvinar to the amygdala is commonly assumed to mediate the non-conscious processing of affective visual stimuli. We review anatomical and physiological data that argue against the notion that such a pathway plays a prominent part in processing affective visual stimuli in humans. Instead, we propose that the primary role of the amygdala in visual processing, like that of the pulvinar, is to coordinate the function of cortical networks during evaluation of the biological significance of affective visual stimuli. Under this revised framework, the cortex has a more important role in emotion processing than is traditionally assumed.


Spatially varying equilibria of mechanical models: application to dermal wound contraction.
Mechanochemical models based on the Oster-Murray continuum framework have been applied to a variety of biological settings to obtain an understanding of the morphogenesis of living tissues. Wound-healing in mammalian skin is an important example, because a complex sequence of biochemical and biomechanical responses are orchestrated to close a wound by a combination of new tissue formation and wound contraction. Mechanical interactions between dermal fibroblastic cells and the collagen-rich extracellular matrix are crucial in the development of a contracted wound state. We and others have previously proposed mechanochemical models for wound repair to gain a greater understanding of both normal and abnormal healing. In the present work, the existence of spatially varying equilibria of these models is investigated by using a small-stain approximation and phase-plane techniques, with numerical simulations to confirm the analytical predictions. These results are sources of novel insight into the roles of key biological parameters in determining the mechanical properties of a contracted wound. These methods may also be relevant to other morphogenetic scenarios for which similar mechanochemical models have been proposed.


Inhibition of TGFBIp expression by lithium: implications for TGFBI-linked corneal dystrophy therapy.
PURPOSE. The purpose of this study was to investigate the effects and molecular mechanisms of lithium on inhibition of TGFBIp expression as a potential therapy for TGFBI-linked corneal dystrophy. METHODS. Primary culture corneal fibroblasts were isolated from the corneas of healthy subjects and patients with granular corneal dystrophy type 2 (GCD2) with a homozygous mutation in TGFBI R124H. Levels of TGFBIp and its mRNA in corneal fibroblasts treated with various lithium (LiCl) concentrations were analyzed by Western blot, RT-PCR, and quantitative real-time PCR. RESULTS. LiCl treatment reduced the expression levels of normal and mutant TGFBIp in a dose- and a time-dependent manner. Furthermore, TGF-β1-induced TGFBIp expression decreased by 35% and 67% after treatment with 5 mM and 10 mM LiCl, respectively. LiCl decreased the level of pSmad3 (S423/425) in a dose-dependent manner. Furthermore, LiCl increased the level of pGSK-3α/β (S21/9) in a dose-dependent manner. Also observed was the interaction between GSK-3β and Smad3, which was enhanced by lithium. In addition, Western blot analysis showed that the ratio of LC3-II/LC3-I in corneal fibroblasts increased after LiCl treatment. Cell viability at different doses was greater than 98%, indicating that LiCl did not induce significant corneal fibroblast death. Finally, the observed attenuating effects of LiCl on TGFBIp expression were not the results of cell death. CONCLUSIONS. The accumulation of mutant TGFBIp ultimately leads to the histopathologic and clinical manifestations associated with TGFBI-linked corneal dystrophy. These data strongly suggest that lithium may be used for the prevention or treatment of this disease.


Excess of non-verbal cases of autism spectrum disorders presenting to orthodox clinical practice in Africa - a trend possibly resulting from late diagnosis and intervention.
OBJECTIVES: Characteristics of children with autism spectrum disorders (ASDs) in Africa are not known because of unavailability of large-scale epidemiological studies in this region. This review explored the age at first presentation to orthodox clinical practice of African children with ASDs and their expressive language ability at presentation. METHODS: A literature search of case series and case reports of ASDs from Africa was done through PubMed/MEDLINE, Google Scholar, African Journals Online (AJOL), and archives of the Nigerian Journal of Psychiatry. Six articles included content relating to age of the child at first presentation to orthodox clinical practice and symptoms at presentation related to expressive language ability and therefore fulfilled the inclusion criteria. Suggestions are made to explain the observations emanating from the review. RESULTS: An excess of non-verbal over verbal cases of ASDs have been presenting to orthodox clinical practice and there is a common denominator of late presentation/diagnosis and in turn late intervention, with most cases presenting for the first time well above 8 years of age. Attempts to explain these observations included low levels of knowledge and awareness about ASDs in Africa; problems with help-seeking behaviour; and lack of mental healthcare facilities and trained personnel. CONCLUSIONS: Enhancement of processes directed at ensuring early diagnosis and interventions, especially interventions aimed at improving speech and language development well and sufficiently early, may bring about a shift in the trend of excess non-verbal cases of ASDs over verbal cases presenting to orthodox clinical practice.


Caspase-2 is required for dendritic spine and behavioural alterations in J20 APP transgenic mice.
Caspases have critical roles in Alzheimer's disease pathogenesis. Here we show that caspase-2 is required for the cognitive decline seen in human amyloid precursor protein transgenic mice (J20). The age-related changes in behaviour and dendritic spine density observed in these mice are absent when they lack caspase-2, in spite of similar levels of amyloid beta (Aβ) deposition and inflammation. A similar degree of protection is observed in cultured hippocampal neurons lacking caspase-2, which are immune to the synaptotoxic effects of Aβ. Our studies suggest that caspase-2 is a critical mediator in the activation of the RhoA/ROCK-II signalling pathway, leading to the collapse of dendritic spines. We propose that this is controlled by an inactive caspase-2/RhoA/ROCK-II complex localized in dendrites, which dissociates in the presence of Aβ, allowing for their activation and entry in the spine. These findings directly implicate caspase-2 as key driver of synaptic dysfunction in Alzheimer's disease and offer novel therapeutic targets.