Osteogenic Differentiation and Biocompatibility of Bovine Teeth Scaffold with Rat Adipose-derived Mesenchymal Stem Cells.
Adipose-derived mesenchymal stem cells (ADMSCs) have great potential for regenerative medicine. These have been combined with biomaterials such as Bovine teeth that are preferred as a periodontal regeneration material. The main purpose of this study is to evaluate and analyze a biocompatibility test and osteogenic differentiation of bovine teeth scaffold seeded with ADMSCs in vitro. A true experimental study with post-test only group design was conducted. Random sampling and Lameshow's formula were used to determine the sample. The scaffold, obtained from bovine teeth as the bone graft material, was analyzed using 3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and its attachment was evaluated by scanning electron microcopy (SEM) and micro-computed tomography with ADMSCs. ADMDSCs attachment present in the bovine teeth scaffold was assessed using SEM at 1-hour, 12-hour, and 24-hour intervals. Analysis of variance was used to analyze the MTT assay results (p < 0.05) based on normality and homogeneity test (p > 0.05). The highest viability of cells (97.08%) was found at a concentration of 10% by means of an MTT test (p < 0.05). The results of three-dimensional bovine teeth scaffold showed the average particle size to be 500 µm. ADMSCs cell attachment to the scaffold bovine teeth showed a significant increase in the number of cells attached after 24 hours compared with those at 1 and 12 hours. Alizarin red staining showed an increase in ADMSC osteogenic differentiation after it was combined with bovine teeth scaffold. The scaffold from bovine teeth is biocompatible and accelerates osteogenic differentiation of ADMSC.


Regulation of mouse chondrocyte differentiation by CCAAT/enhancer-binding proteins.
CCAAT/enhancer-binding protein (C/EBP) β regulates chondrocyte differentiaion and proliferation during endochondral ossification. However, expression and function of other C/EBP family members in chondrocytes have not been fully understood. To understand the comprehensive regulation of chondrocyte differentiation by C/EBPs, we initially examined their expression levels. Among four members (C/EBPα, C/EBPβ, C/EBPδ and C/EBPε) with transactivation domain, expression of Cebpb and Cebpd was abundant compared to Cebpa, while Cebpe was hardly expressed in mouse isolated chondrocytes. Doxycycline (DOX)-inducible overexpression of each of the three C/EBPs (C/EBPα, C/EBPβ and C/EBPδ) in ATDC5 cells suppressed expressions of early differentiation markers including Col2a1, aggrecan and Sox9, enhanced those of late differentiation markers including Mmp13, Vegfa and Col10a1, and decelerated cell proliferation, indicating their overlapped functions in chondrocytes. In contrast, DOX-inducible overexpression of A-CEBP, which exerts a dominant-negative effect against all C/EBPs, increased expressions of early differentiation markers and decreased those of late differentiation markers. Finally, microarray and gene ontology analyses showed that A-CEBP altered many genes related with various events or tissues such as skeletal development, cartilage, cell cycle, inflammation and apoptosis. In conclusion, C/EBPα, C/EBPβ and C/EBPδ regulate proliferation and differentiation of chondrocytes and possibly is involved with apoptosis and inflammation. C/EBPs may play a variety of roles in the homeostasis of joint cartilage under physiological and pathological conditions.


Effect of osteopathic manipulation on gait asymmetry.
Movement and loading asymmetry are associated with an increased risk of musculoskeletal injury, disease progression, and suboptimal recovery. Osteopathic structural screening can be utilized to determine areas of somatic dysfunction that could contribute to movement and loading asymmetry. Osteopathic manipulation treatments (OMTs) targeting identified somatic dysfunctions can correct structural asymmetries and malalignment, restoring the ability for proper compensation of stresses throughout the body. Little is currently known about the ability for OMTs to reduce gait asymmetries, thereby reducing the risk of injury, accelerated disease progression, and suboptimal recovery. To demonstrate whether osteopathic screening and treatment could alter movement and loading asymmetry during treadmill walking. Forty-two healthy adults (20 males, 22 females) between the ages of 18 and 35 were recruited for this prospective intervention. Standardized osteopathic screening exams were completed by a single physician for each participant, and osteopathic manipulation was performed targeting somatic dysfunctions identified in the screening exam. Three-dimensional (3-D) biomechanical assessments, including the collection of motion capture and force plate data, were performed prior to and following osteopathic manipulation to quantify gait mechanics. Motion capture and loading data were processed utilizing Qualisys Track Manager and Visual 3D software, respectively. Asymmetry in the following temporal, kinetic, and kinematic measures was quantified utilizing a limb symmetry index (LSI): peak vertical ground reaction force, the impulse of the vertical ground reaction force, peak knee flexion angle, step length, stride length, and stance time. A 2-way repeated-measures analysis of variance model was utilized to evaluate the effects of time (pre/post manipulation) and sex (male/female) on each measure of gait asymmetry. Gait asymmetry in the peak vertical ground reaction force (-0.6%, p=0.025) and the impulse of the vertical ground reaction force (-0.3%, p=0.026) was reduced in males following osteopathic manipulation. There was no difference in gait asymmetry between time points in females. Osteopathic manipulation did not impact asymmetry in peak knee flexion angle, step length, stride length, or stance time. Among the participants, 59.5% (25) followed the common compensatory pattern, whereas 40.5% (17) followed the uncommon compensatory pattern. One third (33.3%, 14) of the participants showed decompensation at the occipitoatlantal (OA) junction, whereas 26.2% (11), one third (33.3%, 14), and 26.2% (11) showed decompensation at the cervicothoracic (CT), thoracolumbar (TL), and lumbosacral (LS) junctions, respectively. Somatic dysfunction at the sacrum, L5, right innominate, and left innominate occurred in 88.1% (37), 69.0% (29), 97.6% (41), and 97.6% (41) of the participants, respectively. Correcting somatic dysfunction can influence gait asymmetry in males; the sex-specificity of the observed effects of osteopathic manipulation on gait asymmetry is worthy of further investigation. Osteopathic structural examinations and treatment of somatic dysfunctions may improve gait symmetry even in asymptomatic individuals. These findings encourage larger-scale investigations on the use of OMT to optimize gait, prevent injury and the progression of disease, and aid in recovery after surgery.


Dynamic Control of dNTP Synthesis in Early Embryos.
Exponential increase of cell numbers in early embryos requires large amounts of DNA precursors (deoxyribonucleoside triphosphates (dNTPs)). Little is understood about how embryos satisfy this demand. We examined dNTP metabolism in the early Drosophila embryo, in which gastrulation is preceded by 13 sequential nuclear cleavages within only 2 hr of fertilization. Surprisingly, despite the breakneck speed at which Drosophila embryos synthesize DNA, maternally deposited dNTPs can generate less than half of the genomes needed to reach gastrulation. The rest of the dNTPs are synthesized "on the go." The rate-limiting enzyme of dNTP synthesis, ribonucleotide reductase, is inhibited by endogenous levels of deoxyATP (dATP) present at fertilization and is activated as dATP is depleted via DNA polymerization. This feedback inhibition renders the concentration of dNTPs at gastrulation robust, with respect to large variations in maternal supplies, and is essential for normal progression of embryogenesis.


Behavior controlled by scheduled injections of cocaine in squirrel and rhesus monkeys.
Rates and patterns of key-press responding maintained under schedules in which responding resulted in intravenous injections of cocaine were studied in squirrel monkeys and rhesus monkeys. Each injection was followed by a 60- or 100-sec timeout period. Schedule-controlled behavior was obtained at appropriate cocaine doses in each species. Under FR 10 or FR 30 schedules, performance was characterized by high rates of responding (usually more than one response per second) in each ratio. Under FI 5-min schedules, performance was characterized by an initial pause, followed by acceleration of responding to a final rate that was maintained until the end of the interval. Under multiple fixed-ratio fixed-interval schedules, rates and patterns of responding appropriate to each schedule component were maintained. Responding seldom occurred during timeout periods under any schedule studied. At doses of cocaine above or below those that maintained characteristic schedule-controlled behavior, rates of responding were relatively low and patterns of responding were irregular. Characteristic fixed-interval responding was maintained over a wider range of cocaine doses than characteristic fixed-ratio responding. Complex patterns of responding controlled by discriminative stimuli under fixed-ratio or fixed-interval schedules can be maintained by cocaine injections in squirrel monkeys and rhesus monkeys.


Australian doctors are more engaged than UK doctors: why is this the case?
Despite reports highlighting the need for greater medical engagement and the benefits of being widely understood, very little information is available on the status of medical engagement in Australia, and how this compares to the UK. Answering this question will no doubt assist training bodies, curriculum designers and policy makers better understand relevant issues. The medical engagement questionnaire (MES) was emailed to all medical staff working at 159 UK National Health Service Trusts and 18 health service organisations in Australia. The questionnaire consists of 30 predetermined items seeking responses using a 5-point Likert scale. Overall, doctors in the Australian dataset are slightly more engaged, or more positive, than their UK colleagues. Good interpersonal relationships was the only variable that UK doctors scored more positively than their Australian counterparts. At the lower end of the responses, that is the least engaged, we found this even more apparent. Where doctors in Australia are less disengaged, that is still more positive than the UK colleagues. While the profiles of medical engagement vary at the sites and also across the MES and subscales, the data illustrate that overall doctors in Australia feel valued and empowered, and they have purpose and direction and work in a collaborate culture. At the most disengaged end of the scale, Australian doctors are markedly less disengaged than their UK counterparts. There may be numerous factors that influence and change how engaged doctors are in both countries. The most prominent of these are appear to be working conditions and lifestyle, driven by funding and other economics issues. This research is likely to be of great interest to regulators and training bodies in both countries.


Detection and identification of a novel subgroup 16SrII-V phytoplasma associated with Praxelis clematidea phyllody disease.
Praxelis clematidea is a very vigorous non-native weed in tropical and subtropical regions of China. P. clematidea plants showing symptoms of phyllody disease were found in an orchard located in Hainan province, PR China. The presence of phytoplasmas was confirmed by PCR of 16S rRNA gene using phytoplasma universal primers R16mF2/R16mR1 followed by R16F2n/R16R2. According to 16S rRNA gene sequence similarity, the P. clematidea phyllody (PCP) phytoplasma is a 'CandidatusPhytoplasma australasiae'-related strain (99.5 % similarity). The virtual RFLP pattern analyses of 16S rRNA gene sequences indicated that the PCP is a new subgroup within 16 Sr group II. The most similar RFLP pattern is the reference pattern of 16Sr group II, subgroup M, with a similarity coefficient of 0.94. These results were confirmed by phylogenetic analyses of the 16S rRNA gene. These findings suggest that P. clematidea phyllody disease is caused by a new phytoplasma considered to be a novel subgroup, 16SrII-V.


Waking selective neurons in the posterior hypothalamus and their response to histamine H3-receptor ligands: an electrophysiological study in freely moving cats.
Neurons which discharge selectively during waking (waking selective) have been found in the tuberomamillary nucleus (TM) and adjacent areas of the posterior hypothalamus. Although they share some electrophysiological properties with aminergic neurons, there is no direct evidence that they are histaminergic. We have recorded from posterior hypothalamic neurons during the sleep-wake cycle in freely moving cats, and investigated the effects on waking selective neurons of specific ligands of histaminergic H3-receptors, which autoregulate the activity of histaminergic neurons. Two types of neurons were seen. Waking selective neurons, termed "waking-on (W-on)," were located exclusively within the TM and adjacent areas, and discharged at a low regular rate during waking (1.71-2.97 Hz), decreased firing during light slow wave sleep (SWS), became silent during deep SWS and paradoxical sleep (PS) and resumed their activity on, or a few seconds before, awakening. "Waking-related" neurons, located in an area dorsal to the TM, displayed a similar, although less regular, low rate of firing (1.74-5.41 Hz) and a similar discharge profile during the sleep-wake cycle; however, unlike "W-on" neurons, they did not completely stop firing during deep SWS and PS. Intramuscular (i.m.) injection of ciproxifan (an H3-receptor antagonist, 1mg/kg), significantly increased the discharge rate of W-on neurons and induced c-fos expression in histamine-immunoreactive neurons, whereas i.m. injection of imetit (an H3-receptor agonist, 1mg/kg) or microinjection of alpha-methylhistamine (another H3-receptor agonist, 0.025-0.1 microg/0.2 microl) in the vicinity of these cells significantly decreased their discharge rate. Moreover, the effect of the antagonist was reversed by the agonists and vice versa. In contrast, "waking-related" neurons were unaffected by these H3-receptor ligands. These data provide evidence for the histaminergic nature of "W-on" neurons and their role in cortical desynchronization during waking, and highlight the heterogeneity of posterior hypothalamic neuronal populations, which might serve different functions during the wakefulness.


Surgery related complications in percutaneous endoscopic lumbar discectomy under local anesthesia.
The minimally invasive percutaneous endoscopic discectomy (PED) as the postero-lateral approach with the local anesthesia was started in the late 20th century. The procedure only requires 8 mm of skin incision; thus, it is the least invasive disc surgery presently. The surgery related complications were reviewed in the initial 100 cases from the single surgeon (K. S., first author). Two cases showed exiting nerve irritation, and complained of leg paresthetic pain for 6 to 12 weeks after the surgery (2.0%). The symptoms got better with medicines. One showed post-surgical epidural hematoma, and required surgical removal of the mass (1.0%). Two cases complained neck pain during surgery (2.0%). Surgeons would be aware of the specific complications for the postero-lateral approach of PED procedure.


Methods for microbiological quality assessment in drinking water: a comparative study.
The present study aimed to compare several methods for quantifying and discriminating between the different physiological states of a bacterial population present in drinking water. Flow cytometry (FCM), solid-phase cytometry (SPC), epifluorescence microscopy (MSP) and culture method performances were assessed by comparing the results obtained for different water samples. These samples, including chlorinated and non-chlorinated water, were collected in a drinking water treatment plant. Total bacteria were quantified by using SYBR Green II (for FCM) and 4',6'-diamino-2-phenylindole (DAPI) (for MSP), viable and non-viable bacteria were distinguished by using SYBR Green II and propidium iodide dual staining (for FCM), and active cells were distinguished by using CTC (for MSP) and Chemchrome V6 (for FCM and SPC). In our conditions, counts using microscopy and FCM were significantly correlated regarding total bacteria and active cells. Conversely, counts were not significantly similar using solid-phase and FCM for active bacteria. Moreover, the R2A medium showed that bacterial culturability could be recovered after chlorination. This study highlights that FCM appears to be a useful and powerful technique for drinking water production monitoring.


Potential of PEDOT:PSS as a hole selective front contact for silicon heterojunction solar cells.
We show that the highly conductive polymer poly(3,4-ethylenedioxythiophene)-poly(styrenesulfonate) (PEDOT:PSS) can successfully be applied as a hole selective front contact in silicon heterojunction (SHJ) solar cells. In combination with a superior electron selective heterojunction back contact based on amorphous silicon (a-Si), mono-crystalline n-type silicon (c-Si) solar cells reach power conversion efficiencies up to 14.8% and high open-circuit voltages exceeding 660 mV. Since in the PEDOT:PSS/c-Si/a-Si solar cell the inferior hybrid junction is determining the electrical device performance we are capable of assessing the recombination velocity (v I ) at the PEDOT:PSS/c-Si interface. An estimated v I of ~400 cm/s demonstrates, that while PEDOT:PSS shows an excellent selectivity on n-type c-Si, the passivation quality provided by the formation of a native oxide at the c-Si surface restricts the performance of the hybrid junction. Furthermore, by comparing the measured external quantum efficiency with optical simulations, we quantify the losses due to parasitic absorption of PEDOT:PSS and reflection of the device layer stack. By pointing out ways to better passivate the hybrid interface and to increase the photocurrent we discuss the full potential of PEDOT:PSS as a front contact in SHJ solar cells.


Facilitatory Effects of Multi-Word Units in Lexical Processing and Word Learning: A Computational Investigation.
Previous studies have suggested that children and adults form cognitive representations of co-occurring word sequences. We propose (1) that the formation of such multi-word unit (MWU) representations precedes and facilitates the formation of single-word representations in children and thus benefits word learning, and (2) that MWU representations facilitate adult word recognition and thus benefit lexical processing. Using a modified version of an existing computational model (McCauley and Christiansen, 2014), we extract MWUs from a corpus of child-directed speech (CDS) and a corpus of conversations among adults. We then correlate the number of MWUs within which each word appears with (1) age of first production and (2) adult reaction times on a word recognition task. In doing so, we take care to control for the effect of word frequency, as frequent words will naturally tend to occur in many MWUs. We also compare results to a baseline model which randomly groups words into sequences-and find that MWUs have a unique facilitatory effect on both response variables, suggesting that they benefit word learning in children and word recognition in adults. The effect is strongest on age of first production, implying that MWUs are comparatively more important for word learning than for adult lexical processing. We discuss possible underlying mechanisms and formulate testable predictions.


Associations between Cellular Energy and Pediatric Inflammatory Bowel Disease Patient Response to Treatment.
Inflammatory bowel diseases (IBDs), including Crohn's disease (CD) and ulcerative colitis, are chronic diseases of the gastrointestinal tract, with an unknown etiology, that affect over 6.8 million people worldwide. To characterize disease pathogenesis, proteomic and bioinformatic analyses were performed on colon biopsies collected during diagnostic endoscopy from 119 treatment-naïve pediatric patients, including from 78 IBD patients and 41 non-IBD patients who served as controls. Due to the presence of noninflamed and/or inflamed regions in IBD patients, up to two biopsies were obtained from IBD patients as compared to a single noninflamed biopsy from non-IBD pediatric control patients. Additional biopsies were obtained and analyzed from 33 of the IBD patients after IBD-directed therapeutic intervention for comparison of pre- and post-treatment proteomes. SuperSILAC was utilized to perform quantitative analysis of homogenized tissues, which were processed by filter-aided sample preparation. Hierarchical clustering and principal component analyses revealed proteomic patterns that distinguished inflamed from noninflamed tissues independent of therapy. Gene ontology revealed that proteins downregulated in inflammation are associated with metabolism, whereas upregulated proteins contribute to protein processing. A comparison of pre- and post-treatment proteomes from CD patients identified over 100 proteins that are significantly different between patients who responded and those who did not respond to therapy, including creatine kinase B and basigin.


Assessment of hepatic fatty infiltration using spectral computed tomography imaging: a pilot study.
The objective of this study was to investigate the feasibility of using monochromatic spectral computed tomography (CT) imaging to assess fatty infiltration in liver. With spectral CT imaging, phantoms with known fat concentrations were studied for scanning parameter optimization, then 52 patients enrolled into 4 groups (healthy, mild, moderate, and severe fatty infiltration) received abdominal scanning. Based on reconstructed monochromatic images, hepatic attenuation was analyzed, and dual-energy subtraction imaging (DESI) was created for quantifying fat infiltration. Corresponding to various hepatic fat infiltrations, 4 characteristic CT attenuation curve patterns were described. In DESI images, only fat and fatty components appeared bright. For livers without abnormal fat deposition, isolated bright pixels were visualized (% area = 0.5% ± 0.3%). With hepatic fat accumulation increasing, more bright pixels appeared in subtraction images with percentages of total liver area involved in 2.5%, 6.7%, and 13.4% of mild, moderate, and severe fat infiltration cases, respectively (P < 0.05). The corresponding CT values were as follows: 1.33, 2.53, 8.69, and 16.4 Hounsfield units (P < 0.01), which correlated with the % DESI area values (r = 0.9811). Spectral CT imaging is a promising method to quantitatively assess hepatic fat content and fatty infiltration with advantages compared with conventional CT imaging.


Assessment of nitrous oxide production in eutrophicated rivers with inflow of treated wastewater based on investigation and statistical analysis.
Accurate estimation and control of greenhouse gas emissions have been recognized as imperative in recent years. Therefore, frequent investigations under uniform environmental conditions are required to better understand this concept. Thus, six sampling sites with characteristic concentrations of nitrogen and other water quality parameters were selected to investigate the behavior of water quality parameters throughout the year and to statistically examine the correlations among the parameters. Dissolved nitrous oxide (D-N2O) showed the highest positive correlation coefficient with NO2-N among nitrogen species. The results of the principal component analysis suggested that river water quality could be broadly classified based on photosynthesis and contamination from treated wastewater. Photosynthesis caused an increase in pH, with concomitant decrease in D-N2O concentration. Using the results of multiple regression analysis, D-N2O was accurately estimated based on nitrogen concentration, pH, and concentration of organic matter in various situations. The results of a detailed survey suggested that D-N2O was produced in the river from nitrogen sources released from the wastewater treatment plant. The main roles of the wastewater treatment plant for D-N2O behavior in the river were the supply of the nitrogen source that was the precursor of D-N2O, the supply of the nutrients that induced the photosynthesis, and the direct supply of D-N2O at a low water temperature. The models based on multiple regression analysis could efficiently predict the D-N2O concentration produced in rivers at sites downstream of the wastewater treatment plant, except for the direct supply of D-N2O as an effluent at low water temperature.


An Engineered Community Approach for Industrial Cultivation of Microalgae.
Although no species lives in isolation in nature, efforts to grow organisms for use in biotechnology have generally focused on a single-species approach, particularly where a product is required at high purity. In such scenarios, preventing the establishment of contaminants requires considerable effort that is economically justified. However, for some applications in biotechnology where the focus is on lower-margin biofuel production, axenic culture is not necessary, provided yields of the desired strain are unaffected by contaminants. In this article, we review what is known about interspecific interactions of natural algal communities, the dynamics of which are likely to parallel contamination in industrial systems. Furthermore, we discuss the opportunities to improve both yields and the stability of cultures by growing algae in multi-species consortia.


Responses to GABA(A) receptor activation are altered in NTS neurons isolated from chronic hypoxic rats.
The inhibitory amino acid GABA is released within the nucleus of the solitary tract (NTS) during hypoxia and modulates the respiratory response to hypoxia. To determine if responses of NTS neurons to activation of GABA(A) receptors are altered following exposure to chronic hypoxia, GABA(A) receptor-evoked whole cell currents were measured in enzymatically dispersed NTS neurons from normoxic and chronic hypoxic rats. Chronic hypoxic rats were exposed to 10% O(2) for 9-12 days. Membrane capacitance was the same in neurons from normoxic (6.9+/-0.5 pF, n=16) and hypoxic (6.3+/-0.5 pF, n=15) rats. The EC(50) for peak GABA-evoked current density was significantly greater in neurons from hypoxic (21.7+/-2.2 microM) compared to normoxic rats (12.2+/-0.9 microM) (p<0.001). Peak and 5-s adapted GABA currents evoked by 1, 3 and 10 microM were greater in neurons from normoxic compared to hypoxic rats (p<0.05) whereas peak and 5-s adapted responses to 30 and 100 microM GABA were not different comparing normoxic to hypoxic rats. Desensitization of GABA(A)-evoked currents was observed at concentrations greater than 3 microM and, measured as the ratio of the current 5 s after the onset of 100 microM GABA application to the peak GABA current, was the same in neurons from normoxic (0.37+/-0.03) and hypoxic rats (0.33+/-0.04). Reduced sensitivity to GABA(A) receptor-evoked inhibition in chronic hypoxia could influence chemoreceptor afferent integration by NTS neurons.


MIB 1 immunostaining in cervical carcinoma of young patients.
In the present study, we assessed whether biologic characteristics of tumors in young patients differ from those observed in older patients with the same clinical and histologic characteristics, but ranging in age from 50 to 70 years. The hypothesis to be verified was whether cervical carcinoma in young patients presented an increased proliferative activity which might explain more aggressive behavior. Locally advanced cervical carcinoma tumor samples were obtained from our series of patients, maximum age 40 years, and immunohistochemically evaluated by monoclonal MIB 1 antibody (Immunotech, Marseille Cedex, France) on microwave-oven-processed Formalin-fixed paraffin-embedded tissue. Positive staining was expressed as a percentage of positive cells per 10(3) counted neoplastic cells for each case. For each young patient, a control was selected among patients aged >/=50 years (range 50-70) matched for stage, tumor size, histologic type and grading, and lymphvascular invasion. Fourteen of 73 patients (19.2%) with stage I and IIa cervical carcinoma who underwent primary radical surgery at our Institute between 1986 and 1994 were aged </=40 years. The MIB 1 index was significantly higher in young patients with respect to the older group (mean +/- standard deviation, 61.6 +/- 16.3% and 45.2 +/- 12.2%, with P = 0.006). Although any conclusions from this study need to be tempered because of the small number of patients involved, locally advanced cervical carcinomas present in young patients showed a more aggressive biologic behavior, expressed by a higher cell proliferation index.


[Arterioveinous dural malformation in ENT].
The arterioveinous dural malformation (MAVD) is a rare entity between the vascular craneal anomalies with a not well nown ethiology and variability in treatments. We present eleven cases of MAVDs, between them five presented tinnitus as symptom of aparition. The importance of this pathology makes necessary to discard it before a patient consulting because of pulsaltil tinnitus with normal otoscopy.


Effects of ozone on isolated peripheral blood mononuclear cells.
We have investigated the release of cytokines from isolated peripheral human blood mononuclear cells (PBMC) exposed to various ozone concentrations for 10 min and the release of both proinflammatory and immunosuppressive cytokine after 24, 48 and 76 h incubation. Ozonation was performed by exposing for 10 min equal cell numbers and volumes of cell suspension to equal volumes of a gas mixture (1:1 ratio) composed of oxygen-ozone with precise ozone concentrations ranging from 1.0 up to 80 microg/ml (0.02 up to 1.68 mM). Markers of oxidative stress showed a significant relationship between ozone doses and both lipid peroxidation and protein thiol groups content. With the exception of the lowest ozone concentration, the cytokine production of PBMC was depressed particularly at concentrations from 40 mug/ml upwards. There was no significant effect on IL-6 production between exposed or unexposed cells, up to 72 h of incubation. IL-4 production was markedly affected by ozone exposure, showing a marked decrease even at the lowest ozone concentration (2.5 microg/ml) already after 24 h incubation. On the other hand, production of IFN-gamma and TNF-alpha was slightly stimulated by the lowest ozone dose either at all times or only after 72 h incubation, respectively. Analysis of the proliferation index (PI) is consistent with these results showing that, while the lowest concentration stimulates it, progressively increasing O(3) concentrations inhibit the PI. These data show that there is a significant relationship between cytokine production and ozone concentrations and that PBMC are very sensitive to oxidation particularly in presence of serum with low antioxidant capacity.


Proteolytic processing and glycosylation of cathepsin B. The role of the primary structure of the latent precursor and of the carbohydrate moiety for cell-type-specific molecular forms of the enzyme.
The lysosomal cysteine proteinase cathepsin B is synthesized in cultured human hepatoma HepG2 cells as an inactive 44 kDa precursor and subsequently processed to the mature single-chain enzyme with a molecular mass of 33 kDa. Intralysosomal conversion into the two-chain form results in subunits of 27 kDa, 24 kDa (heavy chain) and 5 kDa (light chain). Enzymic deglycosylation reveals that the 27 kDa polypeptide is the glycosylated variant of the carbohydrate-free 24 kDa heavy-chain form. The intracellular transport to the lysosomes is dependent upon mannose 6-phosphate-containing N-linked oligosaccharides. Receptor-mediated endocytosis of human skin-fibroblast-derived procathepsin B by HepG2 cells resulted in processed molecular forms that are not distinguishable from endogenous cathepsin B, thus favouring rather a cell-type-specific processing than structural differences due to the source of the proenzyme. The conversion step of single-chain catehpsin B into the two-chain enzyme is inhibited in vivo by the irreversible cysteine-proteinase inhibitors Z-Phe-Ala-CHN2 and, albeit weaker, Z-Phe-Phe-CHN2. Both substances have no effect on the activation of procathepsin B to the mature enzyme. The carbohydrate moiety of cathepsin B exerts no significant influence on the stability and the enzymatic activity of the enzyme.


Cannabinoid facilitation of fear extinction memory recall in humans.
A first-line approach to treat anxiety disorders is exposure-based therapy, which relies on extinction processes such as repeatedly exposing the patient to stimuli (conditioned stimuli; CS) associated with the traumatic, fear-related memory. However, a significant number of patients fail to maintain their gains, partly attributed to the fact that this inhibitory learning and its maintenance is temporary and conditioned fear responses can return. Animal studies have shown that activation of the cannabinoid system during extinction learning enhances fear extinction and its retention. Specifically, CB1 receptor agonists, such as Δ9-tetrahydrocannibinol (THC), can facilitate extinction recall by preventing recovery of extinguished fear in rats. However, this phenomenon has not been investigated in humans. We conducted a study using a randomized, double-blind, placebo-controlled, between-subjects design, coupling a standard Pavlovian fear extinction paradigm and simultaneous skin conductance response (SCR) recording with an acute pharmacological challenge with oral dronabinol (synthetic THC) or placebo (PBO) 2 h prior to extinction learning in 29 healthy adult volunteers (THC = 14; PBO = 15) and tested extinction retention 24 h after extinction learning. Compared to subjects that received PBO, subjects that received THC showed low SCR to a previously extinguished CS when extinction memory recall was tested 24 h after extinction learning, suggesting that THC prevented the recovery of fear. These results provide the first evidence that pharmacological enhancement of extinction learning is feasible in humans using cannabinoid system modulators, which may thus warrant further development and clinical testing. This article is part of a Special Issue entitled 'Cognitive Enhancers'.


Testing the palindromic target site model for DNA transposon insertion using the Drosophila melanogaster P-element.
Understanding the molecular mechanisms that influence transposable element target site preferences is a fundamental challenge in functional and evolutionary genomics. Large-scale transposon insertion projects provide excellent material to study target site preferences in the absence of confounding effects of post-insertion evolutionary change. Growing evidence from a wide variety of prokaryotes and eukaryotes indicates that DNA transposons recognize staggered-cut palindromic target site motifs (TSMs). Here, we use over 10 000 accurately mapped P-element insertions in the Drosophila melanogaster genome to test predictions of the staggered-cut palindromic target site model for DNA transposon insertion. We provide evidence that the P-element targets a 14-bp palindromic motif that can be identified at the primary sequence level, which predicts the local spacing, hotspots and strand orientation of P-element insertions. Intriguingly, we find that the although P-element destroys the complete 14-bp target site upon insertion, the terminal three nucleotides of the P-element inverted repeats complement and restore the original TSM, suggesting a mechanistic link between transposon target sites and their terminal inverted repeats. Finally, we discuss how the staggered-cut palindromic target site model can be used to assess the accuracy of genome mappings for annotated P-element insertions.


Cadherins at cell-autonomous membrane contacts control macropinocytosis.
Cadherins aggregate and stabilize cell-cell junctions through interactions with adjacent cells. In addition, N-cadherin and E-cadherin concentrate at free edges or at the lamellipodia of migrating cells and are found within large vesicles called macropinosomes, which develop from membrane ruffles. The binding properties of cadherins have not previously been associated with the localization of cadherins at membrane ruffles; however, we report that the dorsal, ventral and lateral membrane contacts that occur as a result of the overlap of membrane ruffles aggregate N-cadherin, and that both N-cadherin and E-cadherin promote macropinosome closure and fluid-phase uptake in macropinosomes. These data reveal a previously unsuspected function for cadherin-mediated cell-cell adhesion molecules in the closure of cell-autonomous membrane contacts at membrane ruffles, resulting in macropinocytosis.


Blood lead levels and risk factors in young children in France, 2008-2009.
The exposure of children to lead has decreased in recent years, thanks notably to the banning of leaded gasoline. However, lead exposure remains a matter of public health concern, because no toxicity threshold has been observed, cognitive effects having been demonstrated even at low levels. It is therefore important to update exposure assessments. A national study was conducted, in 2008-2009, to determine the blood lead level (BLL) distribution in children between the ages of six months and six years in France. We also assessed the contribution of environmental factors. This cross-sectional survey included 3831 children recruited at hospitals. Two-stage probability sampling was carried out, with stratification by hospital and French region. Sociodemographic characteristics were recorded, and blood samples and environmental data were collected by questionnaire. Generalized linear model and quantile regression were used to quantify the association between BLL and environmental risk factors. The geometric mean BLL was 14.9μg/l (95% confidence interval (CI)=[14.5-15.4]) and 0.09% of the children (95% CI=[0.03-0.15]) had BLLs exceeding 100μg/l, 1.5% (95% CI=[0.9-2.1] exceeding 50μg/l. Only slight differences were observed between French regions. Environmental factors significantly associated with BLL were the consumption of tap water in homes with lead service connections, peeling paint or recent renovations in old housing, hand-mouth behavior, passive smoking and having a mother born in a country where lead is often used. In children between the ages of one and six years in France, lead exposure has decreased over the last 15 years as in the US and other European countries. Nevertheless still 76,000 children have BLL over 50μg/l and prevention policies must be pursued, especially keeping in mind there is no known toxicity threshold.


Responses of Picea mariana to elevated CO2 concentration during growth, cold hardening and dehardening: phenology, cold tolerance, photosynthesis and growth.
Seedlings from a northern and a southern provenance of black spruce (Picea mariana Mill. BSP) from eastern Canada were exposed to 37 or 71 Pa of carbon dioxide (CO2) during growth, cold hardening and dehardening in a greenhouse. Bud phenology, cold tolerance and photosynthetic efficiency were assessed during the growing and over-wintering periods. Bud set occurred earlier in elevated [CO2] than in ambient [CO2], but it was later in the southern provenance than in the northern provenance. An increase in seedling cold tolerance in early fall was related to early bud set in elevated [CO2]. Maximal photosystem II (PSII) photochemical efficiency (F(v)/F(m)), effective quantum yield (phi(PSII)), photochemical quenching (q(P)), light-saturated photosynthesis (Amax), apparent quantum efficiency (alpha'), light-saturated rate of carboxylation (Vcmax) and electron transport (Jmax) decreased during hardening and recovered during dehardening. Although Amax and alpha' were higher in elevated [CO2] when measured at the growth [CO2], down-regulation of photosynthesis occurred in elevated [CO2] as shown by lower F(v)/F(m), phi(PSII), Vcmax and Jmax. Elevated [CO2] reduced gene expression of the small subunit of Rubisco and also decreased chlorophyll a/chlorophyll b ratio and nitrogen concentration in needles, confirming our observation of down-regulation of photosynthesis. Elevated [CO2] increased the CO2 diffusion gradient and decreased photorespiration, which may have contributed to enhance Amax despite down-regulation of photosynthesis. Total seedling dry mass was higher in elevated [CO2] than in ambient [CO2] at the end of the growing season. However, because of earlier bud formation and cold hardening, and down-regulation of photosynthesis during fall and winter in elevated [CO2], the treatment difference in dry mass increment was less by the end of the winter than during the growing season. Differences in photosynthetic rate observed during fall, winter and spring account for the inter-annual variations in carbon assimilation of black spruce seedlings: our results demonstrate that these variations need to be considered in carbon budget studies.


Relating instrumental texture, determined by variable-blade and Allo-Kramer shear attachments, to sensory analysis of rainbow trout, Oncorhynchus mykiss, fillets.
Texture is one of the most important quality attributes of fish fillets, and accurate assessment of variation in this attribute, as affected by storage and handling, is critical in providing consistent quality product. Trout fillets received 4 treatments: 3-d refrigeration (R3), 7-d refrigeration (R7), 3-d refrigeration followed by 30-d frozen storage (R3F30), and 7-d refrigeration followed by 30-d frozen storage (R7F30). Instrumental texture of raw and cooked fillets was determined by 3 approaches: 5-blade Allo-Kramer (AK) and variable-blade (VB) attachment with 12 blades arranged in perpendicular (PER) and parallel (PAR) orientations to muscle fibers. Correlation between instrumental texture and sensory hardness, juiciness, elasticity, fatness, and coarseness was determined. Muscle pH remained constant at 6.54 to 6.64. Raw fillets lost 3.66% of their original weight after 30-d frozen storage. After cooking, weight loss further increased to 15.97%. Moisture content decreased from 69.11 to 65.02%, while fat content remained constant at 10.41%. VBPER detected differences in muscle sample strength (P= 0.0019) and demonstrated effect of shear direction reported as maximum force (g force/g sample). AKPER detected differences in energy of shear (g × mm; P= 0.0001). Fillets that received F30 treatments were less extensible. Cooking increased muscle strength and toughness. Force determined by VBPER was correlated with sensory hardness (r= 0.423, P= 0.0394) and cook loss (r= 0.412, P= 0.0450). VB attachment is accurate, valid, and less destructive in fillet texture analysis. A new shearing device was validated with sensory analysis. Settings and parameters obtained could be used to define fillet texture quality associated with muscle fiber orientation.


The anti-herpesvirus activity of (1'S,2'R)-9-[[1',2'-bis(hydroxymethyl)-cycloprop-1'-yl]methyl]guanine is markedly potentiated by the immunosuppressive agent mycophenolate mofetil.
Mycophenolic acid (MPA), the active form of the immunosuppressive agent mycophenolate mofetil (MMF), was found to markedly potentiate the anti-herpesvirus activity of the novel anti-herpesvirus agent A-5021, (1'S,2'R)-9-[[1',2'-bis(hydroxymethyl)cycloprop-1'-yl]methyl]guanine. For example, at a concentration of 1 microg/ml MPA, the activity of A-5021 against HSV-1, HSV-2 and TK(-) HSV-1 increased by a factor of 130, 14 and > or = 189, respectively. Exogenously added guanosine reversed this potentiating effect, suggesting that a depletion of the endogenous dGTP pools enhanced the inhibitory effect of the 5'-triphosphate metabolite of A-5021 on the viral DNA polymerase. The combined effect of A-5021 and MPA on the growth of uninfected Vero cells was additive rather than synergistic. The combination of topically applied MMF (5%) with 0.05% A-5021 (a subactive concentration) completely protected against HSV-1-induced cutaneous lesions in hairless mice, whereas therapy with either compound used alone had no protective effect. These findings may have implications for those transplant recipients that receive MMF as (part of) their immunosuppressive therapy and that develop intercurrent herpesvirus infections for which they need treatment.


Early Inflammatory Cytokine Expression in Cerebrospinal Fluid of Patients with Spontaneous Intraventricular Hemorrhage.
We investigated cerebrospinal fluid (CSF) expression of inflammatory cytokines and their relationship with spontaneous intracerebral and intraventricular hemorrhage (ICH, IVH) and perihematomal edema (PHE) volumes in patients with acute IVH. Twenty-eight adults with IVH requiring external ventricular drainage for obstructive hydrocephalus had cerebrospinal fluid (CSF) collected for up to 10 days and had levels of interleukin-1α (IL-1α), IL-1β, IL-6, IL-8, IL-10, tumor necrosis factor-α (TNFα), and C-C motif chemokine ligand CCL2 measured using enzyme-linked immunosorbent assay. Median [IQR] ICH and IVH volumes at baseline (T0) were 19.8 [5.8-48.8] and 14.3 [5.3-38] mL respectively. Mean levels of IL-1β, IL-6, IL-10, TNF-α, and CCL2 peaked early compared to day 9-10 (p < 0.05) and decreased across subsequent time periods. Levels of IL-1β, IL-6, IL-8, IL-10, and CCL2 had positive correlations with IVH volume at days 3-8 whereas positive correlations with ICH volume occurred earlier at day 1-2. Significant correlations were found with PHE volume for IL-6, IL-10 and CCL2 at day 1-2 and with relative PHE at days 7-8 or 9-10 for IL-1β, IL-6, IL-8, and IL-10. Time trends of CSF cytokines support experimental data suggesting association of cerebral inflammatory responses with ICH/IVH severity. Pro-inflammatory markers are potential targets for injury reduction.


Validation of microscopic observation drug susceptibility testing for rapid, direct rifampicin and isoniazid drug susceptibility testing in patients receiving tuberculosis treatment.
Drug susceptibility testing (DST) is often needed in patients clinically failing tuberculosis (TB) therapy. Most studies of phenotypic direct drug susceptibility tests, such as microscopic observation drug susceptibility (MODS) tests, have been performed in patients not receiving TB treatment. The effect of ongoing TB treatment on the performance of MODS direct DST has not been previously explored, but patients failing such therapy constitute an important target group. The aim of this study was to determine the performance of MODS direct rifampicin and isoniazid DST in patients clinically failing first-line TB treatment, and to compare MODS direct DST with indirect proportion method DST. Sputa from 264 TB patients were cultured in parallel in Lowenstein-Jensen (LJ) and MODS assays; strains were tested for rifampicin and isoniazid susceptibility by the proportion method at the national reference laboratory. Ninety-three samples were culture-positive by LJ and MODS (concordance of 96%; kappa 0.92). With conventional MODS plate DST reading (performed on the same day as the sample is classified as culture-positive), the isoniazid DST concordance was 96.8% (kappa 0.89), and the concordance for rifampicin susceptibility testing was 92.6% (kappa 0.80). Reading of MODS DST plates 1 week after cultures had been determined to be culture-positive improved overall performance marginally-the isoniazid DST concordance was 95.7% (kappa 0.85); and the rifampicin DST concordance was 96.8% (kappa 0.91). Sensitivity for detection of multidrug-resistant TB was 95.8%. MODS testing provided reliable rifampicin and isoniazid DST results for samples obtained from patients receiving TB therapy. A modified DST reading schedule for such samples, with a final reading 1 week after a MODS culture turns positive, marginally improves the concordance with reference DST.


[Two-dimensional surface Raman imaging of a roughened iron electrode in saline solution].
By using a confocal microprobe Raman system and proper surface roughening procedure for iron, the pit corrosion behavior of bare iron electrode in 3.4% NaCl (saline solution) solution has been investigated. The surface-enhanced Raman scattering (SERS) spectra of the corrosion products due to the pit corrosion at positive potential was obtained with high ratio of signal to noise. The application of SERS technique has been extended successfully to the study of iron corrosion. The two-dimensional surface Raman imaging acquired at 660 cm-1 was obtained also with the utility of surface Raman image technique. Results show that different iron oxides should coexist in the pit corrosion region and the distribution of the corrosion products was not uniform.


Chloride currents from the transverse tubular system in adult mammalian skeletal muscle fibers.
Chloride fluxes are the main contributors to the resting conductance of mammalian skeletal muscle fibers. ClC-1, the most abundant chloride channel isoform in this preparation, is believed to be responsible for this conductance. However, the actual distribution of ClC-1 channels between the surface and transverse tubular system (TTS) membranes has not been assessed in intact muscle fibers. To investigate this issue, we voltageclamped enzymatically dissociated short fibers using a two-microelectrode configuration and simultaneously recorded chloride currents (I(Cl)) and di-8-ANEPPS fluorescence signals to assess membrane potential changes in the TTS. Experiments were conducted in conditions that blocked all but the chloride conductance. Fibers were equilibrated with 40 or 70 mM intracellular chloride to enhance the magnitude of inward I(Cl), and the specific ClC-1 blocker 9-ACA was used to eliminate these currents whenever necessary. Voltage-dependent di-8-ANEPPS signals and I(Cl) acquired before (control) and after the addition of 9-ACA were comparatively assessed. Early after the onset of stimulus pulses, di-8-ANEPPS signals under control conditions were smaller than those recorded in the presence of 9-ACA. We defined as attenuation the normalized time-dependent difference between these signals. Attenuation was discovered to be I(Cl) dependent since its magnitude varied in close correlation with the amplitude and time course of I(Cl). While the properties of I(Cl), and those of the attenuation seen in optical records, could be simultaneously predicted by model simulations when the chloride permeability (P(Cl)) at the surface and TTS membranes were approximately equal, the model failed to explain the optical data if P(Cl) was precluded from the TTS membranes. Since the ratio between the areas of TTS membranes and the sarcolemma is large in mammalian muscle fibers, our results demonstrate that a significant fraction of the experimentally recorded I(Cl) arises from TTS contributions.


YY1 modulates the radiosensitivity of esophageal squamous cell carcinoma through KIF3B-mediated Hippo signaling pathway.
Radiotherapy is an important strategy in the comprehensive treatment of esophageal squamous cell carcinoma (ESCC). However, effectiveness of radiotherapy is still restricted by radioresistance. Herein, we aimed to understand the mechanisms underlying ESCC radioresistance, for which we looked into the potential role of YY1. YY1 was upregulated in radioresistant tissues and correlated with poor prognosis of patients with ESCC. YY1 depletion enhanced the radiosensitivity of ESCC in vitro and in vivo. Multi-group sequencing showed that downregulation of YY1 inhibited the transcriptional activity of Kinesin Family Member 3B (KIF3B), which further activated the Hippo signaling pathway by interacting with Integrin-beta1 (ITGB1). Once the Hippo pathway was activated, its main effector, Yes-associated protein 1 (YAP1), was phosphorylated in the cytoplasm and its expression reduced in the nucleus, thus enhancing the radiosensitivity by regulating its targeted genes. Our study provides new insights into the mechanisms underlying ESCC radioresistance and highlights the potential role of YY1 as a therapeutic target for ESCC.


Structure-based optimization of FDA-approved drug methylene blue as a c-myc G-quadruplex DNA stabilizer.
G-quadruplexes are non-canonical DNA secondary structures putatively present in the promoter regions of oncogenes in the human genome. The targeting of promoter G-quadruplex structures to repress oncogene transcription represents a potential anticancer strategy. Here, we have used high-throughput virtual screening to identify FDA-approved drug methylene blue (MB) as a promising scaffold for binding the c-myc oncogene G-quadruplex DNA. Based on molecular docking analysis of MB to the c-myc G-quadruplex, we designed and screened 50 MB derivatives containing side chains that could interact with the G-quadruplex grooves. As a proof-of-concept, the highest-scoring compounds were synthesized and the interactions with the c-myc G-quadruplex were investigated using the FID assay. The results showed that the methylene blue derivatives 6a-c were able to bind to the c-myc G-quadruplex with greater binding affinity compared to the known G-quadruplex binding ligand, crystal violet. The activity of the most potent compound identified from the FID assay, 6b, as an inhibitor for polymerase-drive DNA extension was examined using a PCR-stop assay and compared against that of the parent compound methylene blue. The results of the PCR-stop assay showed that the addition of the side chain improved the activity of the derivatives as an inhibitor compared to the parent compound. The MB derivative 6b was shown to be highly selective towards c-myc G-quadruplex over double-stranded DNA and other biologically relevant G-quadruplexes using UV-visible spectroscopy and mass spectrometry, respectively. The MB derivative 6b could induce or stabilize c-myc G-quadruplex formation in both cell-free and cellular biological models, and displayed higher cytoxicity against human hepatocarcinoma cells compared to the parent compound, MB.


The metamorphosis of amphibian toxicogenomics.
Amphibians are important vertebrates in toxicology often representing both aquatic and terrestrial forms within the life history of the same species. Of the thousands of species, only two have substantial genomics resources: the recently published genome of the Pipid, Xenopus (Silurana) tropicalis, and transcript information (and ongoing genome sequencing project) of Xenopus laevis. However, many more species representative of regional ecological niches and life strategies are used in toxicology worldwide. Since Xenopus species diverged from the most populous frog family, the Ranidae, ~200 million years ago, there are notable differences between them and the even more distant Caudates (salamanders) and Caecilians. These differences include genome size, gene composition, and extent of polyploidization. Application of toxicogenomics to amphibians requires the mobilization of resources and expertise to develop de novo sequence assemblies and analysis strategies for a broader range of amphibian species. The present mini-review will present the advances in toxicogenomics as pertains to amphibians with particular emphasis upon the development and use of genomic techniques (inclusive of transcriptomics, proteomics, and metabolomics) and the challenges inherent therein.


Afferent input from distal forelimb nerves to branching spinoreticular-spinocerebellar neurones in the cat.
Patterns of afferent connections from receptors of the distal forelimb were investigated in neurones located in C6-C7 segments of the spinal cord with branching axons projecting to the lateral reticular nucleus and the cerebellum. Experiments were made on five adult cats under alpha-chloralose anaesthesia. After antidromic identification, EPSPs and IPSPs were recorded from 22 neurones following stimulation of deep radial, superficial radial, median and ulnar nerves. Both excitatory and inhibitory effects were found in the majority of the cells, however, in 2 cases no synaptic actions were recorded. EPSPs were evoked from group I or II muscle, or cutaneous afferents - mostly monosynaptically. IPSPs from muscle, cutaneous or flexor reflex afferents were mostly polysynaptic. Seven various types of convergence were established in the cells investigated. Significance of parallel transmission of integrated information from various receptors of the distal forelimb to the reticular formation and cerebellum is discussed.


[Optimization of the immunocytochemical detection of nestin in paraffin sections of the rat brain].
The aim of this study was to develop an optimal and easy protocol for immunocytochemical demonstration of nestin (an intermediate filament protein which is believed to be a neuronal stem cell marker) in paraffin sections of rat brain with target retrieval procedure being omitted for better preservation of the neural tissue structure. The influence of fixation, character of blocking of nonspecific antibody binding, temperature and time of incubation on the results of immunocytochemical reaction was studied. On the basis of the results obtained, we propose a processing protocol allowing distinct demonstration of nestin in rat brain cells together with the optimal preservation of the nervous tissue structure.


Transcription factors Nrf2 and NF-κB contribute to inflammation and apoptosis induced by intestinal ischemia-reperfusion in mice.
Intestinal ischemia/reperfusion (IIR) is a common pathological event associated with intestinal injury and apoptosis with high mortality. Nuclear factor (NF)-E2-related factor-2 (Nrf2) is a key transcription factor that interacts with NF-κB and has a vital anti-inflammatory effect. However, whether Nrf2 has a role in IIR-induced apoptosis and the possible underlining mechanisms, such as modulation of the inflammation regulation pathway, have remained to be fully elucidated. In the present study, IIR was identified to cause significant intestinal injury and apoptosis, with high expression levels of inflammatory cytokines, as well as the apoptotic proteins B-cell lymphoma 2 (Bcl-2)-associated X protein (Bax) and caspase-3, while simultaneously decreasing the protein levels of Bcl-2. The effect was more pronounced after pretreatment of the animals with all-trans retinoic acid or brusatol, potent inhibitors of Nrf2. t-Butylhydroquinone, an Nrf2 activator, significantly attenuated IIR-induced intestinal injury and apoptosis, with inhibition of the overexpression of the inflammatory cytokines, Bax and caspase-3 protein and partial restoration of Bcl-2 protein expression. Taken together, these results indicated that increased Nrf2 expression reduced IIR-induced intestinal apoptosis and that the protective function of Nrf2 may be based on its anti-inflammatory effects through the inhibition of the NF-κB pathway.


Toxin-induced coagulopathy.
Although warfarin and heparin have been mainstays of anticoagulation for almost 50 years, the recent introduction of multiple oral anticoagulants has led some practitioners to shift away from warfarin as the anticoagulant of choice for various diseases. Major advances have been made in targeting downstream clotting factors in the coagulation cascade, resulting in two major new classes of drugs: direct thrombin inhibitors and factor Xa inhibitors. Developed partially with the patient in mind, these drugs are taken orally and, because of their target specificity, have eliminated the need for routine blood monitoring, making them attractive to patients currently on warfarin.


Receptor for advanced glycation end products: fundamental roles in the inflammatory response: winding the way to the pathogenesis of endothelial dysfunction and atherosclerosis.
The multiligand receptor for advanced glycation end products (RAGE) of the immunoglobulin superfamily is expressed on multiple cell types implicated in the immune-inflammatory response and in atherosclerosis. Multiple studies have elucidated that ligand-RAGE interaction on cells, such as monocytes, macrophages, and endothelial cells, mediates cellular migration and upregulation of proinflammatory and prothrombotic molecules. In addition, recent studies reveal definitive rules for RAGE in effective T lymphocyte priming in vivo. RAGE ligand AGEs may be formed in diverse settings; although AGEs are especially generated in hyperglycemia, their production in settings characterized by oxidative stress and inflammation suggests that these species, in part via RAGE, may contribute to the pathogenesis of atherosclerosis. In murine models of atherosclerosis, vascular inflammation is a key factor and one which is augmented, in parallel with even further increases in RAGE ligands, in diabetic macrovessels. The findings that antagonism and genetic disruption of RAGE in atherosclerosis-susceptible mice strikingly reduces vascular inflammation and atherosclerotic lesion area and complexity link RAGE intimately to these processes and suggest that RAGE is a logical target for therapeutic intervention in aberrant inflammatory mechanisms and in atherosclerosis.


Mitochondrial Cation Signalling in the Control of Inflammatory Processes.
Mitochondria are the bioenergetic organelles responsible for the maintenance of cellular homeostasis and have also been found to be associated with inflammation. They are necessary to induce and maintain innate and adaptive immune cell responses, acting as signalling platforms and mediators in effector responses. These organelles are also known to play a pivotal role in cation homeostasis as well, which regulates the inflammatory responses through the modulation of these cation channels. In particular, this review focuses on mitochondrial Ca2+ and K+ fluxes in the regulation of inflammatory response. Nevertheless, this review aims to understand the interplay of these inflammation inducers and pathophysiological conditions. In detail, we discuss some examples of chronic inflammation such as lung, bowel, and metabolic inflammatory diseases caused by a persistent activation of the innate immune response due to a dysregulation of mitochondrial cation homeostasis.


[Relationship of the clinical efficiency of tocilizumab therapy to the serum level of matrix metalloproteinase-3 in patients with rheumatoid arthritis].
To evaluate the impact of tocilizumab (TCZ) therapy on the level of matrix metalloproteinase-3 (MMP-3) 4, 24, and 48 weeks after treatment initiation in relation to the clinical efficiency of TCZ therapy by the Disease Activity Score (DAS28), the Clinical Disease Activity Index (CDAI), and the Simplified Disease Activity Index (SDAI). Forty-two rheumatoid arthritis (RA) patients who had received 6 intravenous infusions of TCZ 8 mg/kg at a 4-week interval during permanent therapy with disease-modifying anti-rheumatic drugs (DMARD) and glucocorticosteroids (GCS) were examined. Then TCZ was discontinued and the patients continued to receive the previous therapy with DMARD and GCS. The European League Against Rheumatism (EULAR) classification criteria, as well as SDAI and CDAI were used to evaluate the efficiency of TCZ therapy. The serum concentration of MMP-3 was measured by enzyme immunoassay using the test systems (Invitrogen, USA). After 24 weeks of TCZ therapy (at 48 weeks following trial initiation), DAS28 was 4.69 (3.86; 5.44); the SDAI of 17.8 (10.7; 29.5) and the CDAI of 17.1 (7.2; 26.2) corresponded to moderate disease activity. At 48 weeks, DAS28 remission (< 2.6 scores) remained in 5 (11.90%) patients; SDAI (< or = 3.3 scores) and CDAI (< or = 2.8 scores) remissions did in 3 (7.1%) and 4 (9.5%) patients, respectively. There was a significant reduction in MMP-3 concentrations at 4, 24, and 48 weeks of the therapy, which was 61, 73, and 49.40% of the baseline level. ROC analysis indicated that the normalization of MMP-3 levels in RA patients at 24 weeks of TCZ therapy (a cut-off < or =16.5 ng/ml) was associated with the maintenance of remission/low disease activity from SDAI and CDAI 24 weeks after the drug use (the area under the receiver operating curve was 0.762; 95% confidence interval: 0.548-0.976). Analysis of the results of 48-week TCZ therapy suggests its ability to reduce the levels of markers of bone and cartilage destruction in patients with RA. Serum MMP-3 determination at 24 weeks of therapy may be useful in predicting the maintenance of remission/low activity from SDAI and CDAI after discontinuation of the drug.


Outcomes After the Surgical Evacuation of Traumatic Acute Subdural Hematomas: The tASDH Risk Score.
Acute subdural hematoma (ASDH) is a common pathology following traumatic brain injury (TBI). There is sparse data on the prediction of clinical outcomes following traumatic ASDH (tASDH) evacuation. We investigated prognosticators of outcome following evacuation of tASDHs, with subset analysis in a cohort of octogenarians. We developed a scoring system for stratifying the risk of in-hospital mortality for patients undergoing tASDH evacuation. A retrospective chart review was performed to identify all patients who underwent tASDH evacuation. Baseline clinical and demographic data including age, traumatic brain injury mechanism, admission Glasgow Coma Scale (GCS), and Rotterdam computed tomography Scale (RCS) were collected. In-hospital outcomes such as mortality and discharge disposition were collected. A scoring system (tASDH Score) which incorporates RCS (1-2 points), admissions GCS (0-1 points), and age (0-1 point) was created to predict the risk of in-hospital mortality following tASDH evacuation. Being an octogenarian (OR = 6.91 [2.20-21.71], P = 0.0009), having a GCS of 9-12 (OR = 1.58 [1.32-4.12], P = 0.027) or 3-8 (OR = 2.07 [1.41-10.38], P = 0.018), and having an RCS of 4-6 (OR = 3.49 [1.45-8.44], P = 0.0055) were independently predictive of in-hospital mortality. The in-hospital mortality rate was lower for those with a tASDH score of 1 (10%), compared to those with a score of 2 (12%), 3 (42%), and 4 (100%). Octogenarians with an RCS of 4-6 and an admission GCS <13 have a high risk of mortality following tASDH evacuation. Knowledge of which patients are unlikely to survive ASDH evacuation may help guide neurosurgeons in prognostication and goals of care discussions.


Cenegermin in Pediatric Neurotrophic Keratopathy.
We report a case of neurotrophic keratopathy (NK) arising after surgery for rhabdomyosarcoma of the jaw successfully treated with Cenegermin eye drops (Oxervate; Dompé Farmaceutici, Milan, Italy) in a 3-year-old boy. At the age of 1 year, the boy underwent surgery for rhabdomyosarcoma of the jaw, followed by radiotherapy. Subsequent NK was initially treated with preservative-free artificial tears, topical combination of dexamethasone 0.1% and netilmicin 0.3% (Netildex; Sifi, Catania, Italy), and moxifloxacin 0.5% (Vigamox; Alcon, Fort Worth, TX), followed by 10 cycles of a topical eye biopolymer containing a poly-carboxymethyl glucose sulfate solution (Cacicol; Theà, Clermont-Ferrand, France) and 4 amniotic membrane transplantations. Keratopathy was recalcitrant to therapy and tissue transplant. Therapy was switched to Cenegermin eye drops 6 times daily for 8 weeks. Complete healing of the corneal epithelium was achieved at 3 weeks into treatment. Cenegermin was effective in restoring corneal integrity in this pediatric patient with NK.


Nudges, shoves and budges: Behavioural economic policy frameworks.
Behavioural economics-the study of human decision making and how it sometimes deviates systematically from the assumptions of standard economic theory-has attracted a lot of attention in the health policy discourse over recent years. Many appear to believe that behavioural economic findings can be used only to help inform policies that manipulate the choices made by citizens, ie, the so-called nudge policy. However, these findings can be used to inform several different policy frameworks, from seemingly innocuous liberty-preserving changes to the contexts people operate in, to the outlawing of certain corporate behaviours. This article depicts diagrammatically, with the aid of a "behavioural policy cube" and in relation to smoking cessation interventions, the conceptual parameters of several behavioural economic-informed policy frameworks, which could be easily extended to other areas of health, and indeed broader public, policy.


Differences in Stress and Anxiety Among Women With and Without Children in the Household During the Early Months of the COVID-19 Pandemic.
The COVID-19 pandemic and resulting lockdowns have had a disproportionate impact on parents of children under 18, particularly women. Mandatory school closures and loss of childcare resulted in parents balancing work, teaching, and childcare needs. A number of studies have examined changes in mental health of parents, but to date no studies have compared the differences in stress and anxiety levels between women with and without children in the United States. Adult women from the Washington State Twin Registry (WSTR) (N = 1,014, pair N = 529) and mothers of twin children enrolled in the WSTR (N = 147) completed an online survey examining several health-related behaviors and outcomes and their self-reported changes due to COVID-19. We conducted two studies to examine the impact of children on stress and anxiety levels among women. In study 1, we assessed whether women living in households with children under the age of 18 have higher levels of stress and anxiety than those without children in their household. We found that perceived stress levels did not differ between women with and without children in the household, but anxiety levels were higher among women living with children than those without. In study 2, we assessed whether the correlation between children in the household and stress/anxiety is accounted for by non-random genetic and environmental selection effects, causal processes, or both using a sample of adult female twins. We found that the presence of children in the household was associated with higher levels of stress and anxiety. However, this association is confounded by genetic and shared environmental factors. Our findings highlight the need to provide supporting resources to women living with children in the household during and after the COVID-19 pandemic.


"Time is My Own Treasure": Parental Autonomy Support and Academic Procrastination Among Chinese Adolescents.
Previous studies have shown that academic procrastination not only affects middle school students' academic performance but also causes them physical and psychological stress, as well as negative emotions. Therefore, it is necessary to explore the influencing factors of academic procrastination and its internal mechanisms. This study aims to explore the relationship between perceived parental autonomy support and academic procrastination, as well as the role of autonomous motivation and time management disposition. Cross-sectional data from 662 middle school students were collected using the Parental Autonomy Support Scale, Academic Procrastination Inventory for Middle School Students, Academic Self-Regulation Questionnaire and Adolescence Time Management Disposition Scale. SPSS and its PROCESS macro were used for data analysis. After gender and age were controlled, the results showed that perceived parental autonomy support could not only directly predict middle school students' academic procrastination but also predict three paths of procrastination: (1) the mediating role of autonomous motivation, (2) the mediating role of time management disposition, and (3) the chain mediating role of autonomous motivation and time management disposition. Autonomous motivation and time management disposition played a chain mediating role in the relationship between perceived parental autonomy support and academic procrastination in middle school students.


Transdermal delivery of valsartan: I. Effect of various terpenes.
The objective of the present study was to investigate the effect of various terpenes, including a diterpene, forskolin (FSK; a putative penetration enhancer), on skin permeation of valsartan. Permeation studies were carried out with Automated Transdermal Diffusion Cells Sampling System (SFDC 6, LOGAN Instruments Corp., NJ, USA) through rat skin and human cadaver skin (HCS) using ethanol: IPB (pH 7.4) (40:60) as vehicle. The efficacy of the study terpenes for permeation of valsartan across rat skin and human cadaver skin was found in the order of cineole > d-limonene > l-menthol > linalool > FSK and cineole > d-limonene > linalool > l-menthol > FSK, respectively. No apparent skin irritation (erythema, edema) was observed on treatment of skin with terpenes including FSK. FT-IR, DSC, and histopathological studies revealed that FSK enhanced the skin permeation of the active drug by disruption and extraction of lipid bilayers of SC in consonance with other terpenes.


Nonlinear dynamic buckling of stiffened plates under in-plane impact load.
This paper presents a simple solution of the dynamic buckling of stiffened plates under in-plane impact loading. Based on large deflection theory, a discretely stiffened plate model has been used. The tangential stresses of stiffeners and in-plane displacement are neglected. Applying the Hamilton's principle, the motion equations of stiffened plates are obtained. The deflection of the plate is taken as Fourier series, and using Galerkin method the discrete equations can be deduced, which can be solved easily by Runge-Kutta method. The dynamic buckling loads of the stiffened plates are obtained form Budiansky-Roth criterion.


Action potentials in a giant algal cell: a comparative approach to mechanisms and evolution of excitability.
The giant alga Chara corallina generates action potentials (APs) in response to mechanical stimulation, injury, or direct electrical stimulation. Students examine the waveform characteristics of these APs using standard intracellular recording techniques. Intracellular recording is easier than with neurons because of the large size of the Chara cell. Students observe very negative resting potentials (up to -250 mV), large AP amplitudes with depolarizing peaks approaching 0 mV, AP durations of seconds, and refractory periods up to several minutes. Students calculate Nernst potentials for the ions distributed across the Chara cell membrane to hypothesize the ions responsible for the resting potential and for the depolarizing phase of the AP. These calculations suggest that K(+) is responsible for the resting potential and that Ca(2+) influx and Ca(2+)-activated Cl(-) efflux are responsible for depolarizing phases of the AP, which they are. Comparison of the Chara AP characteristics with animal neuron and muscle APs reinforces understanding of mechanisms of excitability in animals, demonstrates that multiple solutions exist for action potential generation, and leads to discussion of the evolution of ion channels and excitability.


Developing a Risk Governance Framework on Radiological Emergency, Preparedness, and Response for Emergency Responders: Protocol for a Mixed Methods Study.
Risk governance involves processes and mechanisms to understand how risk decisions are taken and executed. This concept has gained a reputation over time as being essential for emerging comprehensive management that defines the success of an organization. While guiding documents that explain the use of risk management related to nuclear safety and security are available worldwide, few locally conducted studies have explained risk governance practices in areas where hazard usage is known, such as in radiological emergencies. This paper describes a protocol that was used to determine several factors that influence emergency responders' perceptions toward radiological risk practices and visualize the risk radiological framework for emergency preparedness and response. A mixed methods study with a convergent design was performed. A qualitative analysis was performed using a case study approach where 6 key informants were purposely sampled for in-depth interview, and a cross-sectional study involving a self-administered questionnaire was conducted among approximately 260 emergency respondents from national regulatory, research, and services organizations. NVivo (version 12, QSR International) was used to analyze the interview transcripts and emerging themes were identified through abductive coding. Simultaneously, multiple logistic regression analysis was used to determine significant predictors that form the equation model. The study is still underway. Qualitative findings were based on transcript-coding that informed the relevant thematic analysis, while statistical analyses including multiple logistic regression analysis measured the adjusted odds ratio of significant variables for the equation model. The study is expected to conclude in late 2021. Important emerging themes and significant factors that are related to the emergency responders' perceptions regarding radiological governance practices were determined through the convergent design. This potentially facilitated the development of a plausible radiological risk governance framework. Furthermore, our results will provide key insights that can be used in future studies. DERR1-10.2196/25877.


Olfactory sexual inhibition and the westermarck effect.
The Westermarck effect (sexual inhibition among individuals raised together) is argued to be mediated olfactorily. Various animals, including humans, distinguish among individuals by scent (significantly determined by MHC genotype), and some avoid cosocialized associates on this basis. Possible models of olfactory mechanisms in humans are evaluated. Evidence suggests aversions develop during an early sensitizing period, attach to persons as much as to their scents, and are more powerful among females than among males. Adult to child aversions may develop similarly, but more likely result from stimulus generalization. This hypothesis accords with current evidence and yields testable predictions (e.g., anosmia will prevent inhibition) that, should they be supported, will conclusively ground the Westermarck effect in a biological mechanism.


On the reconstitution of the crystalline components of the sea urchin fertilization membrane.
The characteristics of the reconstitution of a crystalline component of the sea urchin fertilization membrane are presented. The reassembly of large aggregates of cylindrical or tubular components is effected by the addition of calcium or other divalent cations. The reassembly requires a slightly alkaline pH and is little affected by increasing ionic strength. Reassembly is strongly inhibited by treatment with reducing agents such as dithiothreitol. The role of this protein in the formation of the fertilization membrane and its possible relation to the calcium-insoluble proteins of the mitotic apparatus are discussed.


Kindlin-2 regulates colonic cancer stem-like cells survival and self-renewal via Wnt/β-catenin mediated pathway.
Cancer Stem Cells (CSCs) have emerged as a critical mediator in recurrence and resistance in cancers. Kindlin-isoform (1 and 2) binds with cytoplasmic β-tail of integrin and are essential co-activators of integrin function. Given their important function in regulating cancer hallmarks such as cell proliferation, invasion, migration, and metastasis, we hypothesize that it might play a critical role in CSC growth, survival, and self-renewal of colon cancer. Using knockdown approaches, we inhibited Kindlin-2 expression in HCT116 and HT29 colon cancer cells. Extreme limiting dilution and self-renewal assay were performed to measure the role of Kindlin in colonic CSC. Standard methods such as qRT-PCR and western blotting were carried out to understand the signaling cascade by which Kindlin regulates CSC marker expression and downstream targets. Our data show isoform-specific upregulation of Kindlin-2 in colonic CSCs. The silencing of Kindlin-2 reduces colonosphere formation, decreases CSC size, and self-renewal marker genes such as CD-133, CXCR-4, LGR-5, and C-MYC. Kindlin-2 silencing reduces colonosphere proliferation, invasion, and migration of colonic CSCs. Mechanistically, Kindlin-2 silencing reduces the expression, and nuclear localization of β-catenin, and decreases β-catenin target genes such as C-MYC, cyclin D1, DKK-1, and Snail-1. Our study delineates the isoform-specific activity of Kindlin-2 in regulating Colonic CSC. Isoform-specific targeting of Kindlin-2 may be a novel strategy to tackle this devastating disease.


Local fertilizers to achieve food self-sufficiency in Africa.
One of the key Sustainable Development Goals (SDG) set by the United Nations (UN) aims by 2030 to "end hunger, achieve food security and improved nutrition and promote sustainable agriculture". Fertilizers will play a pivotal role in achieving that goal given that ~90% of crop production growth is expected to come from higher yields and increased cropping intensity. However, materials-science research on fertilizers has received little attention, especially in Africa. In this work we present an overview of the use of fertilizers in Africa to date, and based on that overview we suggest future research directions for material scientists. Developing a new generation of local and affordable fertilizers will launch Africa into a new phase of remunerative agricultural production that in turn will lead to both food self-sufficiency and considerable progress towards goals of food and nutrition security.


Ovarian response and follow-up outcomes in women diagnosed with cancer having fertility preservation: Comparison of random start and early follicular phase stimulation - cohort study.
To determine response to controlled ovarian stimulation in a random start cycle and utilisation of cryopreserved oocytes and embryos in cancer patients. A retrospective cohort study was carried out in an assisted reproductive treatment centre. Participants included 137 cancer patients who underwent controlled ovarian stimulation for fertility preservation between 1 Feb 2003 and 30 June 2016. The primary outcome variable was number of oocytes retrieved. Multivariable logistic regression analysis was performed, and differences compared using Chi squared test and student t-test as appropriate. P < 0.05 was considered statistically significant. Using the antagonist protocol, there was no difference in number of oocytes retrieved between the early follicular phase or at random start stimulation; 11.9 (95% CI 10.3-13.5) and 12.9 (95% CI 9.6-16.2), P = 0.602, respectively. Similarly, the number of embryos frozen was comparable between those starting stimulation in early follicular and random phase, 6.7 (95% CI 5.7-7.7) and 5.1 (95% CI 3.6-6.5), P= 0.1508 respectively. Among patients undergoing fertility preservation, those who returned to attempt a pregnancy had an ongoing pregnancy rate of 24.3%. Overall, 65% of oocytes and embryos were still in storage, however, 16 (11.7%) had elected to have their oocytes or embryos disposed of. For women faced with potential gonadotoxic treatment and requiring urgent fertility preservation, ovarian stimulation with the antagonist protocol can be started at random without compromising ovarian response. Pregnancy rates following utilisation of frozen-thawed oocytes and embryos are promising, however, more research is needed to understand reasons underlying disposition of oocytes and embryos especially when survival following cancer treatment has improved significantly.


A novel mode of action of an ArfGAP, AMAP2/PAG3/Papa lpha, in Arf6 function.
Previously we reported that AMAP2/PAG3/Papalpha/KIAA0400, a GTPase-activating protein (GAP), acts to antagonize Arf6 function when overexpressed, whereas it was shown to exhibit efficient GAP activities for other Arf isoforms in vitro. Here, we found that AMAP2, through its ArfGAP domain, binds to GTP-Arf6 but not to GDP-Arf6 or other Arfs irrespective of nucleotide status. The majority of AMAP2 was localized to intracellular tubulovesicular structures and redistributed to Arf6-enriched membrane areas upon Arf6 activation. In HeLa cells, Arf6 has been shown to be involved in the clathrin-independent endocytosis of Tac, but not the clathrin-dependent endocytosis of transferrin. We found that Arf6 silencing inhibited the internalization of Tac, but not transferrin, in HeLa cells. Internalization of Tac, but not transferrin, was also significantly inhibited by AMAP2 silencing and overexpression. AMAP2 was moreover found to bind to amphiphysin IIm, a component of the endocytic machinery, via its proline-rich domain. We propose that AMAP2 has dual mechanisms for its function; it exhibits efficient catalytic GAP activity for the class I and II Arfs and yet is involved in the cellular function of the class III Arf without immediate GAP activity. These dual mechanisms of AMAP2 may be important for the cellular function of GTP-Arf6.


Late primary graft dysfunction after lung transplantation and bronchiolitis obliterans syndrome.
Primary graft dysfunction (PGD) is a common early complication after lung transplantation. We conducted a retrospective cohort study of 334 recipients to evaluate the impact of PGD graded at 24, 48 and 72 h on the risk of bronchiolitis obliterans syndrome (BOS) development (stage 1) and progression (stages 2 and 3). We constructed multivariable Cox proportional hazards models to determine the risk of BOS attributable to PGD in the context of other potential risk factors including acute rejection, lymphocytic bronchitis and respiratory viral infections. All grades of PGD at all time points were significant risk factors for BOS development and progression independent of acute rejection, lymphocytic bronchitis and respiratory viral infections. Specifically, PGD grade 1 at T24 was associated with a relative risk of BOS stage 1 of 1.93, grade 2 with a relative risk of 2.29 and grade 3 with a relative risk of 3.31. Furthermore, this direct relationship between the severity of PGD and the risk of BOS persisted at all time points. We conclude that all grades of PGD at all time points are independent risk factors for BOS development and progression. Future strategies that might attenuate the severity of PGD may mitigate the risk of BOS.


Imaia, a new truffle genus to accommodate Terfezia gigantea.
Originally described from Japan by Sanshi Imai in 1933, the hypogeous ascomycete Terfezia gigantea was subsequently discovered in the Appalachian Mountains of the USA. Morphological, electron microscopic, and phylogenetic studies of specimens collected in both regions revealed that, despite this huge geographic disjunction, (1) the Japanese and Appalachian specimens are remarkably similar both in morphology and the sampled rDNA sequences, (2) the species unambiguously falls into the Morchellaceae and is separated from the genus Terfezia in the Pezizaceae, (3) its spores are much larger than those of Terfezia spp. and are enclosed in a unique, electron-semitransparent, amorphous epispore that appears to be permeated with minute, meandering strands or canals. In addition to the molecular phylogenetic results, the numerous nuclei in ascospores, the dome shaped, striate ascus septal plugs and the long cylindric Woronin bodies also strengthen the family assignment to the Morchellaceae. Moreover, the species occurs in moist, temperate forests as opposed to the xeric to arid habitats of other Terfezia spp. We propose the new, monotypic genus Imaia to accommodate the species.


An electron-microscope study of naturally occurring and cultured cells of Neisseria Gonorrhoeae.
Electron-microscope studies are reported in which gonococci grown in cultures are compared with those present in urethral exudates. Whereas naturally occurring cells presented a smooth appearance with parallel cell layers, cultured cells were rough and their surface layers appeared to be disorganised. Three different types of pili were observed in cultures; the most common type seemed to be a product of unfavourable growth conditions. The number of piliate cells in pus was variable but always much lower than the number of non-piliate cells found. A study of free endotoxin present in cultures was made and its fine structure is described. Further observations suggest that although the majority of gonococci are killed after phagocytosis, others survive and multiply--giving rise to clusters of gonococci within remnants of phagocytes. It is suggested that these coated clusters are infectious units.


Comparison of Multiple Chronic Obstructive Pulmonary Disease (COPD) Indices in Chinese COPD Patients.
Chronic obstructive pulmonary disease (COPD) is a serious chronic condition with a global impact. Symptoms of COPD include progressive dyspnea, breathlessness, cough, and sputum production, which have a considerable impact on the lives of patients. In addition to the human cost of living with COPD and the resulting death, COPD entails a huge economic burden on the Chinese population, with patients spending up to one-third of the average family income on COPD management in some regions is clinically beneficial to adopt preventable measures via prudent COPD care utilization, monetary costs, and hospitalizations. Toward this end, this study compared the relative effectiveness of six indices in predicting patient healthcare utilization, cost of care, and patient health outcome. The six assessment systems evaluated included the three multidimensional Body mass index, Obstruction, Dyspnea, Exercise capacity index, Dyspnea, Obstruction, Smoking, Exacerbation (DOSE) index, and COPD Assessment Test index, or the unidimensional measures that best predict the future of patient healthcare utilization, cost of care, and patient health outcome among Chinese COPD patients. Multiple linear regression models were created for each healthcare utilization, cost, and outcome including a single COPD index and the same group of demographic variables for each of the outcomes. We conclude that the DOSE index facilitates the prediction of patient healthcare utilization, disease expenditure, and negative clinical outcomes. Our study indicates that the DOSE index has a potential role beyond clinical predictions.


Smaller left ventricular end-systolic diameter and lower ejection fraction at baseline associated with greater ejection fraction improvement after revascularization among patients with left ventricular dysfunction.
To investigate the predictive roles of pre-operative left ventricular (LV) size and ejection fraction (EF) in EF improvement and outcome following revascularization in patients with coronary artery disease (CAD) and LV dysfunction. Revascularization may improve EF and long-term outcomes of patients with LV dysfunction. However, the determinants of EF improvement have not yet been investigated comprehensively. Patients with EF measurements before and 3 months after revascularization were enrolled in a cohort study (No. ChiCTR2100044378). All patients had baseline EF ≤ 40%. EF improvement was defined as absolute increase in EF > 5%. According to LV end-systolic diameter (LVESD) (severely enlarged or not) and EF (≤35% or of 36-40%) at baseline, patients were categorized into four groups. A total of 939 patients were identified. A total of 549 (58.5%) had EF improved. Both LVESD [odds ratio (OR) per 1 mm decrease, 1.05; 95% CI, 1.04-1.07; P < 0.001] and EF (OR per 1% decrease, 1.06; 95% CI, 1.03-1.10; P < 0.001) at baseline were predictive of EF improvement after revascularization. Patients with LVESD not severely enlarged and EF ≤ 35% had higher odds of being in the EF improved group in comparison with other three groups both in unadjusted and adjusted analysis (all P < 0.001). The median follow-up time was 3.5 years. Patients with LVESD not severely enlarged and EF ≤ 35% had significantly lower risk of all-cause death in comparison with patients with LVESD severely enlarged and EF ≤ 35% [hazard ratio (HR), 2.73; 95% CI, 1.28-5.82; P = 0.009], and tended to have lower risk in comparison with patients with LVESD severely enlarged and EF of 36-40% (HR, 2.00; 95% CI, 0.93-4.27; P = 0.074). Among CAD patients with reduced EF (≤ 40%) who underwent revascularization, smaller pre-operative LVESD and lower EF had greatest potential to have EF improvement and better outcome. Our findings imply the indication for revascularization in patients with LV dysfunction who presented with lower EF but smaller LV size.


Motion-based microwave tomographic measurement device for three-dimensional coverage in a magnetic resonance system.
We have developed a fully 3D data acquisition system for microwave breast imaging which can operate simultaneously inside a magnetic resonance imaging (MRI). MRI is used regularly for breast imaging to distinguish tumors from normal tissue. It generally has poor specificity unless used with a gadolinium contrast agent. Microwave imaging could fill this need because of the good endogenous tumor:normal tissue property contrast, especially in light of safety concerns for gadolinium. The antenna array consists of 16 monopole antennas positioned in a horizontal circle surrounding the breast which can then be moved vertically for 3D coverage of the breast. The tank system materials were chosen to minimize artifacts in the MR image within the specific shared imaging zone. The support rods are stainless steel, albeit positioned sufficiently far from the imaging target to have little effect. The mechanical motion parts are all 3D printed plastic. Unlike many conventional antennas, the monopoles consist of just the center conductor and insulator of the coaxial cable, making it one of the least possible metallic structures. Data were acquired both inside and outside of the MR bore to confirm that the MR bore did not have adverse effects on the microwave imaging process. The imaging tank was filled with a mixture of glycerin and water to both provide a reasonable property match to the phantom and to highly attenuate the fields which also acted to suppress multi-path signals. Microwave images were reconstructed using our Gauss-Newton scheme combined with a log transformation for a more linear convergence. MR images were also acquired to assess the effects of the microwave tank structures on the imaging. The microwave measurement data were acquired in log magnitude and phase format at 200 MHz increments from 700-1900 MHz. Each antenna acted sequentially as a transmitter while the complement of 15 acted as a receiver. The single frequency images were reconstructed using a Gauss-Newton iterative technique with a standard log transformation to linearize the process. The data showed that the signal strengths were between 7-10 dB lower for the case when the array was inside the MRI versus when not. Notwithstanding, the image quality was still high because of the significant signal to noise ratio. The reconstructed images in both situations demonstrated good 3D object recovery of the vertically size and shaped varying object. The MR images were not adversely affected by the presence of antennas or feed structures. We have demonstrated that our technique can recover high-quality images of a 3D varying object within an MRI system. Compatibility issues have been addressed for both the microwave and MRI systems. The reduced SNR for the case operating in the MRI did not adversely affect the images. To the best of our knowledge, this is the first example of a microwave imaging system operating in an MRI with full 3D volumetric capability.


Quantitative dot blot analysis (QDB), a versatile high throughput immunoblot method.
Lacking access to an affordable method of high throughput immunoblot analysis for daily use remains a big challenge for scientists worldwide. We proposed here Quantitative Dot Blot analysis (QDB) to meet this demand. With the defined linear range, QDB analysis fundamentally transforms traditional immunoblot method into a true quantitative assay. Its convenience in analyzing large number of samples also enables bench scientists to examine protein expression levels from multiple parameters. In addition, the small amount of sample lysates needed for analysis means significant saving in research sources and efforts. This method was evaluated at both cellular and tissue levels with unexpected observations otherwise would be hard to achieve using conventional immunoblot methods like Western blot analysis. Using QDB technique, we were able to observed an age-dependent significant alteration of CAPG protein expression level in TRAMP mice. We believe that the adoption of QDB analysis would have immediate impact on biological and biomedical research to provide much needed high-throughput information at protein level in this "Big Data" era.


Assessment of prevalence and distribution of spotted fever group rickettsiae in Manitoba, Canada, in the American dog tick, Dermacentor variabilis (Acari: Ixodidae).
Little is known about the distribution and prevalence of the spotted fever group rickettsiae in Canada. We conducted active surveillance for tick-associated rickettsiae in 10 localities in Manitoba. A total of 1044 adult American dog ticks, Dermacentor variabilis (Acari: Ixodidae), were collected and screened for spotted fever group rickettsiae. Rickettsia montanensis was the only species of rickettsia detected. The mean prevalence of infection was 9.8% (range, 0.00-21.74% among localities). The proportion of infected male and female ticks was not significantly different; however, tick populations near the northern limit of D. variabilis distribution in Manitoba had a lower prevalence of infection compared to tick populations from more southern localities in the province.


[Contribution of rennin-angiotensin-aldosterone system genes and NO synthase gene to the development of arterial hypertension].
Arterial hypertension (AH) is one of the most widespread cardiovascular disorders, 39.2% of men and 41.1% of women having elevated arterial pressure (AP). Hence, the necessity to elucidate possible causes of this abnormality. Heredity is considered to be a major factor determining AP in humans, and researchers all over the world are engaged in the search for AP markers. This paper is focused on genes controlling the renin-angiotensin-aldosterone system, viz. genes of angiotensin II, type 1 angiotensin II receptors, angiotensin-converting enzyme, and NO synthase. An overview of population studies with special reference to these genes indicates that molecular-genetic mechanisms of AH remain unclear. Joint efforts of researchers working in different centres are needed to address the problem.