Molecular characterization of three novel splicing mutations causing factor V deficiency and analysis of the F5 gene splicing pattern.
Factor V deficiency is a rare autosomal recessive hemorrhagic disorder, associated with bleeding manifestations of variable severity. In the present study, we investigated the molecular basis of factor V deficiency in three patients, and performed a comprehensive analysis of the factor V gene (F5) splicing pattern. Mutational screening was performed by DNA sequencing. Wild-type and mutant F5 mRNA were expressed by transient transfection in COS-1 cells, followed by reverse-transcriptase polymerase chain reaction and sequencing. Real-time reverse-transcriptase polymerase chain reaction was used to evaluate degradation of mRNA carrying premature termination codons. Mutational screening identified three hitherto unknown splicing mutations (IVS8+6T>C, IVS21+1G>A, and IVS24+1_+4delGTAG). Production of mutant transcripts in COS-1 cells demonstrated that both IVS21+1G>A and IVS24+1_+4delGTAG cause the activation of cryptic donor splice sites, whereas IVS8+6T>C causes exon-8 skipping (F5-Delta 8-mRNA). Interestingly, F5-Delta 8-mRNA was also detected in wild-type transfected samples, human liver, platelets, and HepG2 cells, demonstrating that F5 exon-8 skipping takes place physiologically. Since F5-Delta 8-mRNA bears a premature termination codons, we investigated whether this transcript is subjected to nonsense-mediated mRNA decay degradation. The results confirmed the involvement of nonsense-mediated mRNA decay in the degradation of F5 PTC(+) mRNA. Moreover, a comprehensive analysis of the F5 splicing pattern led to the identification of two in-frame splicing variants resulting from skipping of exons 3 and 5-6. The functional consequences of three splicing mutations leading to FV deficiency were elucidated. Furthermore, we report the identification of three alternatively spliced F5 transcripts.


Oscillations and patterns in spatially discrete models for developmental intercellular signalling.
We extend previous models for nearest neighbour ligand-receptor binding to include both lateral induction and inhibition of ligand and receptor production, and different geometries (strings of cells and hexagonal arrays, in addition to square arrays). We demonstrate the possibility of lateral inhibition giving patterns with a characteristic length scale of many cell diameters, when receptor production is included. In contrast, lateral induction combined with inhibition of receptor synthesis cannot give rise to a patterning instability under any circumstances. Interesting new dynamics include the analytical prediction and consequent numerical observation of spatiotemporal oscillations, this depends crucially on the production terms and on the relationship between the decay rates of ligand and free receptor. Our approach allows for a detailed comparison with the model for Delta-Notch interactions of Collier et al. [4], and we find that a formal reduction may be made only when the ligand receptor binding kinetics are very slow. Without such very slow receptor kinetics, spatial pattern formation via lateral inhibition in hexagonal cellular arrays requires significant activation of receptor production, a feature that is not apparent from previous analyses.


Control of the anisotropic shape of cobalt nanorods in the liquid phase: from experiment to theory… and back.
The polyol process is one of the few methods allowing the preparation of metal nanoparticles in solution. Hexagonal close packed monocrystalline Co nanorods are easily obtained in basic 1,2-butanediol at 448 K after a few minutes using a Co(II) dicarboxylate precursor. By using a combined experimental and theoretical approach, this study aims at a better understanding of the growth of anisotropic cobalt ferromagnetic nanoparticles by the polyol process. The growth of Co nanorods along the c axis of the hexagonal system was clearly evidenced by transmission electron microscopy, while the mean diameter was found to be almost constant at about 15 nm. Powder X-ray diffraction data showed that metallic cobalt was generated at the expense of a non-reduced solid lamellar intermediate phase which can be considered as a carboxylate ligand reservoir. Density functional theory calculations combined with a thermodynamic approach unambiguously showed that the main parameter governing the shape of the objects is the chemical potential of the carboxylate ligand: the crystal habit was deeply modified from rods to platelets when increasing the concentration of the ligand, i.e. its chemical potential. The approach presented in this study could be extended to a large number of particle types and growth conditions, where ligands play a key role in determining the particle shape.


Spatiotemporal Asymmetry in Metachronal Rowing at Intermediate Reynolds Numbers.
In drag-based swimming, individual propulsors operating at low Reynolds numbers (where viscous forces dominate over inertial forces) must execute a spatially asymmetric stroke to produce net fluid displacement. Temporal asymmetry (that is, differing duration between the power vs. recovery stroke) does not affect the overall generated thrust in this time-reversible regime. Metachronal rowing, in which multiple appendages beat sequentially, is used by a wide variety of organisms from low to intermediate Reynolds numbers. At the upper end of this range, inertia becomes important, and increasing temporal asymmetry can be an effective way to increase thrust. However, the combined effects of spatial and temporal asymmetry are not fully understood in the context of metachronal rowing. To explore the role of spatiotemporal asymmetry in metachronal rowing, we combine laboratory experiments and reduced-order analytical modeling. We measure beat kinematics and generated flows in two species of lobate ctenophores across a range of body sizes, from 7 to 40 mm in length. We observe characteristically different flows in ctenophores of differing body size and Reynolds number, and a general decrease in spatial asymmetry and increase in temporal asymmetry with increasing Reynolds number. We also construct a one-dimensional mathematical model consisting of a row of oscillating flat plates whose flow-normal areas change with time, and use it to explore the propulsive forces generated across a range of Reynolds numbers and kinematic parameters. The model results show that while both types of asymmetry increase force production, they have different effects in different regions of the parameter space. These results may have strong biological implications, as temporal asymmetry can be actively controlled while spatial asymmetry is likely to be partially or entirely driven by passive fluid-structure interaction.


Euthanasia and the healthcare professional.
Euthanasia, often called mercy killing, is the act of inducing death often on terminally ill individuals for reasons assumed to be merciful. Euthanasia can be active or passive, and voluntary or involuntary. Voluntary euthanasia indicates the patient's full consent to terminate life, whereas involuntary euthanasia indicates the lack of patient's consent. Active euthanasia means the termination of life using an agent such as a drug, whereas passive euthanasia refers to the withholding a potentially life-prolonging treatment. Euthanatizing agents terminate life through various mechanisms, causing physical damage, direct or indirect tissue hypoxia, depression of the nervous system. Euthanasia is an issue filled with moral and ethical, as well as legal dilemmas that cannot be avoided. As members of the health care team, pharmacists may be asked to provide advice involving some form of euthanasia.


Characterization of Cryptococcus neoformans variety gattii SOD2 reveals distinct roles of the two superoxide dismutases in fungal biology and virulence.
We studied superoxide dismutases (SODs) in the encapsulated yeast Cryptococcus neoformans (Cn) variety gattii to analyse the role of mitochondrial MnSOD (SOD2) in fungal biology and virulence. SOD2 was cloned from a Cn cosmid library, sod2 mutant and sod2 + SOD2 reconstituted strains were constructed by homologous recombination, and two sod1sod2 double mutants were constructed by replacing SOD2 in the sod1 mutant with the sod2::HYG allele. The SOD2 protein (SOD2p) encoded 225 amino acids, with 36-66% identity with other fungal SOD2ps. SOD2 deletion rendered Cn highly growth-defective at 37 degrees C in 19-20% oxygen (normal air), and this defect was reversed by limiting oxygen to 1.3% as well in the presence of antioxidant, ascorbic acid. The sod2 mutant accumulated significantly more reactive oxygen species (ROS) at 37 degrees C as well at 30 degrees C in the presence of antimycin A, suggesting that SOD2p is the primary defence of Cn against the superoxide anion (O(2) (.-)) in the mitochondria. The sod2 was also highly susceptible to redox-cycling agents, high salt and nutrient limitations. The sod2 mutant was avirulent in intranasally infected mice and markedly attenuated in its virulence in intravenously infected mice. The virulence defect of sod2 mutant appeared related to its growth defects in high oxygen environment, but not resulting from increased sensitivity to oxidative killing by phagocytes. The sod1sod2 double mutants were avirulent in mice. Additionally, sod1sod2 double mutants showed a marked reduction in the activities of other known Cn virulence factors; and they were more susceptible to PMN killing than was the sod2 single mutant. Previously, we reported that the attenuation of sod1 mutant in mice was resulting from enhanced susceptibility to phagocyte killing, combined with a reduction in the activities of a number of virulence factors. Thus, SOD1p and SOD2p play distinct roles in the biology and virulence of Cn var. gattii via independent modes of action.


Predicting Adolescents' Bullying Participation from Developmental Trajectories of Social Status and Behavior.
The aim of this study was to determine how trajectory clusters of social status (social preference and perceived popularity) and behavior (direct aggression and prosocial behavior) from age 9 to age 14 predicted adolescents' bullying participant roles at age 16 and 17 (n = 266). Clusters were identified with multivariate growth mixture modeling (GMM). The findings showed that participants' developmental trajectories of social status and social behavior across childhood and early adolescence predicted their bullying participant role involvement in adolescence. Practical implications and suggestions for further research are discussed.


Pregnancy induces numerical and functional changes of CD4+CD25 high regulatory T cells in patients with rheumatoid arthritis.
In a prospective study we investigated whether numerical and functional changes of CD4+CD25(high) regulatory T cells (Treg) were associated with changes of disease activity observed during pregnancy and post partum in patients with rheumatoid arthritis (RA). The frequency of CD4+CD25(high) T cells was determined by flow cytometry in 12 patients with RA and 14 healthy women during and after pregnancy. Fluorescence-activated cell sorting (FACS) was used to sort CD4+CD25(high) T cells and CD4+CD25- T cells were stimulated with anti-CD3 and anti-CD28 monoclonal antibodies alone or in co-culture to investigate proliferation and cytokine secretion. Frequencies of CD4+CD25(high) Treg were significantly higher in the third trimester compared to 8 weeks post partum in patients and controls. Numbers of CD4+CD25(high) Treg inversely correlated with disease activity in the third trimester and post partum. In co-culture experiments significantly higher amounts of IL10 and lowered levels of tumour necrosis factor (TNF)alpha and interferon (IFN)gamma were found in supernatants of the third trimester compared to postpartum samples. These findings were independent from health or disease in pregnancy, however postpartum TNFalpha and IFN gamma levels were higher in patients with disease flares. The amelioration of disease activity in the third trimester corresponded to the increased number of Treg that induced a pronounced anti-inflammatory cytokine milieu. The pregnancy related quantitative and qualitative changes of Treg suggest a beneficial effect of Treg on disease activity.


Influenza A (H1N1) patients admitted to intensive care units during the 2009 pandemics: clinical features and outcomes.
To describe the clinical features and outcomes of patients admitted with influenza A (H1N1) infection in two private hospitals' intensive care units in São Paulo, Brazil, during the 2009 pandemics. A retrospective cohort study was conducted to evaluate demographic data, initial clinical presentation, prognostic scores [Simplified Acute Physiology Score (SAPS) 3 and Sequential Organ Failure Assessment (SOFA)], comorbidities, outcomes and treatment of patients with confirmed pandemic influenza diagnosis from July to September 2009. 22 patients were admitted. Median age was 30 (25-43.5) years. Median SAPS 3 and SOFA were 42 (37-49) and 2 (1-3.5), respectively. Comorbidities were common (45.4%), especially obesity (22.7%). Two (9.1%) patients were pregnant. Five (22.7%) patients required invasive mechanical ventilation, with high positive end expiratory pressures (median of 16 cmH2O, interquartile range 10-25cmH2O). There was a 50% incidence of non-invasive ventilation failure. Most (77.2%) of patients were treated with oseltamivir. Hospital mortality was 4.5%. Initial SAPS 3, SOFA and PaO2/FiO2 ratio were associated with mechanical ventilation requirement (p<0.01). Pandemic influenza infection mainly affected young and obese patients. In this study, patients were less severe than those previously described, what explains our low mortality and mechanical ventilation needs. However, high positive end expiratory pressures were required for mechanically ventilated patients.


Evaluation of coronary microvascular function and nitric oxide synthase intron 4a/b polymorphism in patients with coronary slow flow.
Slow coronary flow (SCF) is reported to be associated with increased risk of cardiovascular disease. We have used coronary flow reserve measurement by transthoracic Doppler echocardiography to determine coronary microvascular function in patients with SCF and to determine whether the intron 4a/b polymorphism of the eNOS gene influences coronary endothelial function. Overall, 96 patients with SCF and 79 controls were enrolled in the study. Coronary flow was quantified according to the thrombolysis in myocardial infarction (TIMI) frame count (TFC) on angiogram. Coronary diastolic peak flow velocities (DPFV) were measured with color Doppler flow mapping at baseline and after dipyridamole infusion. Coronary flow reserve was calculated as the ratio of hyperemic to baseline DPFV. The eNOS 4a/b polymorphism was detected by PCR. Patients with diabetes were excluded from the study. The SCF group was comparable to the control group in terms of demographic and clinical characteristics, except for hemoglobin and HDL-cholesterol levels, TFC of the left anterior descending artery, the circumflex artery, and the right coronary artery; the mean TFC was higher in the SCF group. Hyperemic DPFV and the hyperemic/baseline DPFV ratio were significantly lower in the SCF group when compared with the control group. However, baseline DPFV were similar in both groups. The number of patients with eNOS4 a/a and eNOS4 a/b phenotypes was statistically higher in SCF groups. The frequency of allele 'a' of the eNOS4 gene was also statistically higher in the SCF group. When patients were grouped according to the presence or absence of allele 'a' of the eNOS4 gene, statistically significant differences were found in the TFC of the left anterior descending artery, the circumflex artery; mean TFC; baseline DPFV; and hyperemic/baseline DPFV. Univariate analysis in which eNOS4 b/b was used as the referent group showed that the presence of allele 'a' of the eNOS4 gene significantly predicted SCF (odds ratio: 2.79, 95% confidence interval: 1.32-5.89; P=0.007). In multivariate analysis using a model adjusted for variables with a P value lower than 0.10 in univariate analyses, the presence of allele 'a' of the eNOS4 gene was found to be an independent predictor of SCF (odds ratio: 3.22, 95% confidence interval: 1.28-8.82; P=0.013). The presence of allele 'a' may be a risk factor for microvascular endothelial dysfunction and higher TFCs in SCF patients.


Periapical cemental dysplasia.
This case report shows the evolution of periapical cemental dysplasia (PCD) lesions over a 13-year period, and demonstrates that PCD lesions mature over time from purely radiolucent to radiopaque.


Current results of left ventricular assist device therapy in France: the ASSIST-ICD registry.
Our goal was to provide a picture of left ventricular assist device (LVAD) activity in France between 2007 and 2016 based on the multicentric ASSIST-ICD registry. We retrospectively collected 136 variables including in-hospital data, follow-up survival rates and adverse events from 671 LVAD recipients at 20 out of 24 LVAD implant centres in France. The average follow-up time was 1.2 years (standard deviation: 1.4); the total follow-up time was 807.5 patient-years. The included devices were the HeartMate II®, HeartWare LVAS® or Jarvik 2000®. The overall likelihood of being alive while on LVAD support or having a transplant (primary end point) at 1, 2, 3 and 5 years postimplantation was 65.2%, 59.7%, 55.9% and 47.7%, respectively, given a cumulative incidence of 29.2% of receiving a transplant at year 5. At implantation, 21.5% of patients were on extracorporeal life support. The overall rate of cardiogenic shock at implantation was 53%. The major complications were driveline infection (26.1%), pump pocket or cannula infection (12.6%), LVAD thrombosis (12.2%), ischaemic (12.8%) or haemorrhagic stroke (5.4%; all strokes 18.2%), non-cerebral haemorrhage (9.1%) and LVAD exchange (5.2%). The primary end point (survival) was stratified by age at surgery and by the type of device used, with inference from baseline profiles. The primary end point combined with an absence of complications (secondary end point) was also stratified by device type. The ASSIST-ICD registry provides a real-life picture of LVAD use in 20 of the 24 implant centres in France. Despite older average age and a higher proportion of patients chosen for destination therapy, survival rates improved compared to those in previous national registry results. This LVAD registry contrasts with other international registries because patients with implants have more severe disease, and the national policy for graft attribution is distinct. We recommend referring patients for LVAD earlier and suggest a discussion of the optimal timing of a transplant for bridged patients (more dismal results after the second year of support?).


Statistical analysis of the parameters of a neuro-genetic algorithm.
Interest in hybrid methods that combine artificial neural networks and evolutionary algorithms has grown in the last few years, due to their robustness and ability to design networks by setting initial weight values, by searching the architecture and the learning rule and parameters. This paper presents an exhaustive analysis of the G-Prop method, and the different parameters the method requires (population size, selection rate, initial weight range, number of training epochs, etc.) are determined. The paper also the discusses the influence of the application of genetic operators on the precision (classification ability or error) and network size in classification problems. The significance and relative importance of the parameters with respect to the results obtained, as well as suitable values for each, were obtained using the ANOVA (analysis of the variance). Experiments show the significance of parameters concerning the neural network and learning in the hybrid methods. The parameters found using this method were used to compare the G-Prop method both to itself with other parameter settings, and to other published methods.


Molecular Landscape of Therapy-related Myeloid Neoplasms in Patients Previously Treated for Gynecologic and Breast Cancers.
Definition of therapy-related myeloid neoplasms (TRMN) is only based on clinical history of exposure to leukemogenic therapy. No specific molecular classification combining therapy-related acute myeloid leukemia and therapy-related myelodysplastic syndromes has been proposed. We aimed to describe the molecular landscape of TRMN at diagnosis, among 77 patients with previous gynecologic and breast cancer with a dedicated next-generation sequencing panel covering 74 genes. We investigated the impact of clonal hematopoiesis of indeterminate potential-associated mutations (CHIP-AMs defined as presence at TRMN stage of mutations described in CHIP with a frequency >1%) on overall survival (OS) and the clinical relevance of a modified genetic ontogeny-based classifier that categorized patients in 3 subgroups. The most frequently mutated genes were TP53 (31%), DNMT3A (19%), IDH1/2 (13%), NRAS (13%), TET2 (12%), NPM1 (10%), PPM1D (9%), and PTPN11 (9%). CHIP-AMs were detected in 66% of TRMN patients, with no impact on OS. Yet, patients with CHIP-AM were older and had a longer time interval between solid tumor diagnosis and TRMN. According to our modified ontogeny-based classifier, we observed that the patients with TP53 or PPM1D mutations had more treatment lines and complex karyotypes, the "MDS-like" patients were older with more gene mutations, while patients with "De novo/pan-AML" mutations were younger with more balanced chromosomal translocations. Median OS within each subgroup was 7.5, 14.5, and 25.2 months, respectively, with statistically significant difference in multivariate analysis. These results support the integration of cytogenetic and molecular markers into the future TRMN classification to reflect the biological diversity of TRMN and its impact on outcomes.


The impact of preoperative magnetic resonance images on outcome of cemented vertebrae.
Refracture of cemented vertebrae is often seen after percutaneous vertebroplasty. The purpose of this prospective study was to evaluate pre-procedural magnetic resonance images (MRI) for the prediction of further collapse and vertebral height loss after vertebroplasty. This study included 81 consecutive patients (73 women and 8 men) with osteoporotic compression fractures. MR studies were performed 1-5 days before vertebroplasty. Patients were followed to evaluate refracture for a minimum of 6 months after treatment. Cox proportional hazards model was used to evaluate relationships between clinical data, covariates on pre-procedural MRI, and the presence of cemented vertebrae refracture. The mean refracture rate was estimated with the Kaplan-Meier method. After a mean follow-up of 23.0 ± 8.2 months, 46 cemented vertebrae (57%) experienced refracture, and the mean loss of anterior vertebral height was 11.3%. The 1-year refracture rate after vertebroplasty was 7%, and rapid increased to 76% in the third year. Cox proportional analysis showed that any 1% decrease in signal intensity on T2-weighted images of the injured vertebra will increase the refracture rate by 0.74% (OR = 0.26, 95% CI 0.08-0.81, p = 0.02), and a 1% increase in the poorly enhanced volume ratio will increase the refracture rate by 4.3% (OR = 5.32, 95% CI 1.22-23.14, p = 0.03). Quantitative pre-procedural MRI appears to be useful in exploring vertebrae with poor bone marrow integrity, which effectively predicts the subsequent refracture of cemented vertebra.


Single agent paclitaxel as a first-line therapy in advanced urothelial carcinoma: its efficacy and safety in patients even with pretreatment renal insufficiency.
Cisplatin-based chemotherapy is the mainstay of the treatment for advanced urothelial cancer, but patients with renal insufficiency before therapy are usually contraindicated to receiving platinum-based chemotherapy. Paclitaxel is one of the most promising agents against advanced urothelial carcinoma in recent trials and it can be easily tolerated even in patients with compromised renal function. We conducted a study in order to evaluate the efficacy and safety of paclitaxel as a first-line therapy in advanced urothelial carcinoma patients. Thirteen advanced chemo-naive urothelial carcinoma patients with a median age of 71 years were studied, seven of them demonstrating renal insufficiency (pretreatment serum creatinine > or = 1.5 mg/dl). All 13 patients received a minimum of two cycles of paclitaxel 175 mg/m2, delivered by intravenous infusion for 3 h every 3 weeks. Four of the 13 patients responded to treatment, a response rate of 30.8%, with two of these achieving complete remission and two showing partial responses. The median overall survival period of all 13 patients was nine months (95% Cl: 6.51-11.49) and our study revealed a statistical tendency in the difference of median overall survival time between responders and non-responders (13 months versus 7.5 months, log-rank p = 0.038), although the number of cases was limited. The differences in response rate and median overall survival time, comparing patients with renal insufficiency and those with normal renal function, were not significant. Treatment-related toxicity was mild, with only two (15.4%) patients suffering from grade 3-4 leukopenia. No treatment-related mortality was noted. Single-agent paclitaxel can be used as a first-line therapy in advanced urothelial carcinoma patients, and is especially suitable for those with pretreatment renal insufficiency, since the antitumor activity is significant while toxicity is well tolerated.


Impacts of neonicotinoid seed treatments on soil-dwelling pest populations and agronomic parameters in corn and soybean in Quebec (Canada).
Agricultural soil pests, including wireworms (Coleoptera: Elateridae), are managed primarily with pesticides applied directly to seeds before sowing. Seeds coated with neonicotinoids have been used widely in Quebec (Canada) for several years. To assess the agronomic and economic value of neonicotinoid seed treatments in soybeans and corn in Quebec, trials were conducted from 2012 to 2016 in 84 fields across seven regions in Quebec. We evaluated the effect of neonicotinoid seed treatments on soil pest densities, crop damage and yield. The results showed that 92.6% of corn fields and 69.0% of soybean fields had less than 1 wireworm per bait trap. However, no significant differences in plant stand or yield were observed between treated and untreated corn or soybeans during the study. This study shows that neonicotinoid seed treatments in field crops in Quebec are useful in less than 5% of cases, given the very low level of pest-associated pressure and damage, and that they should not be used prophylactically. Integrated pest management (IPM) strategies need to be developed for soil insect pests to offer effective alternative solutions to producers.


The Life Experiences Among Primary Family Caregivers of Home-Based Palliative Care.
An increasing number of patients with terminal illnesses prefer to die in their own homes due to aging, high medical payments, a limited number of hospitalization days, and the ability to receive care from family members. However, few studies have been conducted on the subjective perception and value of caregivers for home-based palliative care (HBPC). To identify common themes and topics of primary family caregivers' lived experiences with HBPC when taking care of terminally ill family members. We conducted audio-recorded transcripts of one-on-one in-depth interviews of primary family caregivers of HBPC. Through a purposive sampling method, the participants were all interviewed; these interviews were transcribed verbatim and analyzed using a grounded theory approach. A total of 22 primary family caregivers participated in the study. "Wholeheartedly accompanying one's family to the end of life at home" was the core category. Six main themes describing caregivers' experiences emerged from the interviews: (1) learning the basic skills of end-of-life home care, (2) arranging the sharing and rotation of care, (3) preparing for upcoming deaths and funerals, (4) negotiating the cultural and ethical issues of end-of-life home care, (5) ensuring a comfortable life with basic life support, and (6) maintaining care characterized by concern, perseverance, and patience. Primary family caregivers of HBPC need support and must learn home care skills by means of the holistic approach. It is crucial to establish assessment tools for caregivers' preparedness for HBPC, including biopsychosocial and cultural considerations.


Differential modulation of repetitive firing and synchronous network activity in neocortical interneurons by inhibition of A-type K(+) channels and Ih.
GABAergic interneurons provide the main source of inhibition in the neocortex and are important in regulating neocortical network activity. In the presence 4-aminopyridine (4-AP), CNQX, and D-APV, large amplitude GABAA-receptor mediated depolarizing responses were observed in the neocortex. GABAergic networks are comprised of several types of interneurons, each with its own protein expression pattern, firing properties, and inhibitory role in network activity. Voltage-gated ion channels, especially A-type K(+) channels, differentially regulate passive membrane properties, action potential (AP) waveform, and repetitive firing properties in interneurons depending on their composition and localization. HCN channels are known modulators of pyramidal cell intrinsic excitability and excitatory network activity. Little information is available regarding how HCN channels functionally modulate excitability of individual interneurons and inhibitory networks. In this study, we examined the effect of 4-AP on intrinsic excitability of fast-spiking basket cells (FS-BCs) and Martinotti cells (MCs). 4-AP increased the duration of APs in both FS-BCs and MCs. The repetitive firing properties of MCs were differentially affected compared to FS-BCs. We also examined the effect of Ih inhibition on synchronous GABAergic depolarizations and synaptic integration of depolarizing IPSPs. ZD 7288 enhanced the amplitude and area of evoked GABAergic responses in both cell types. Similarly, the frequency and area of spontaneous GABAergic depolarizations in both FS-BCs and MCs were increased in presence of ZD 7288. Synaptic integration of IPSPs in MCs was significantly enhanced, but remained unaltered in FS-BCs. These results indicate that 4-AP differentially alters the firing properties of interneurons, suggesting MCs and FS-BCs may have unique roles in GABAergic network synchronization. Enhancement of GABAergic network synchronization by ZD 7288 suggests that HCN channels attenuate inhibitory network activity.


Expression of nuclear receptors of gingiva in polycystic ovarian syndrome: a preliminary case study.
Oestrogen is mainly responsible for alterations in blood vessels and progesterone stimulates the production of inflammatory mediators. In females, during puberty, ovulation and pregnancy, there is an increase in the production of sex steroid hormones, which results in increased gingival inflammation, characterized by gingival enlargement, increased bleeding and crevicular fluid flow. This article presents a case of a patient who presented with a complaint of gingival swelling and spontaneous bleeding that persisted for more than two months. Her health history documented the recently diagnosed presence of polycystic ovarian syndrome. Clinical examination revealed enlarged painful gingival tissues, which bled when touched. After completion of Phase I therapy, the enlargement did not subside and a biopsy sample was taken. This was compared with another patient who had the same health condition but did not show any gingival enlargement. Testing of tissue samples for oestrogen and progesterone receptors showed the first patient to be positive for oestrogen receptors but negative for progesterone, whereas the control was negative for both. Positive oestrogen receptors suggest that polycystic ovarian syndrome has some effect on the periodontium. The dental consequences of this condition, highly prevalent among young females, are typically ignored. Further studies warrant establishment of a clinical association and future diagnosis.


Intramural esophageal hematoma after cardioversion.
Intramural esophageal hematoma is a very rare condition. We report a case of a 40-year-old male, presenting with retrosternal pain and dysphagia. On the day before admission tarry stool and minimal vomiting of old blood was noticed. Diagnostic procedures showed an intramural esophageal hematoma, which had developed 2 weeks and 3 days after cardioversion and anticoagulation therapy. Etiology, differential diagnosis, the diagnostic approach and a brief review of the literature are discussed.


Errors, near misses and adverse events in the emergency department: what can patients tell us?
We sought to determine whether patients or their families could identify adverse events in the emergency department (ED), to characterize patient reports of errors and to compare patient reports to events recorded by health care providers. This was a prospective cohort study in a quaternary care inner city teaching hospital with approximately 40,000 annual visits. ED patients were recruited for participation in a standardized interview within 24 hours of ED discharge and a follow-up interview 3-7 days after discharge. Responses regarding events were tabulated and compared with physician and nurse notations in the medical record and hospital event reporting system. Of 292 eligible patients, 201 (69%) were interviewed within 24 hours of ED discharge, and 143 (71% of interviewees) underwent a follow-up interview 3-7 days after discharge. Interviewees did not differ from the base ED population in terms of age, sex or language. Analysis of patient interviews identified 10 adverse events (5% incident rate; 95% confidence interval [CI] 2.41%-8.96%), 8 near misses (4% incident rate; 95% CI 1.73%-7.69%) and no medical errors. Of the 10 adverse events, 6 (60%) were characterized as preventable (2 raters; kappa=0.78, standard error [SE] 0.20; 95% CI 0.39-1.00; p=0.01). Adverse events were primarily related to delayed or inadequate analgesia. Only 4 out of 8 (50%) near misses were intercepted by hospital personnel. The secondary interview elicited 2 out of 10 adverse events and 3 out of 8 near misses that had not been identified in the primary interview. No designation (0 out of 10) of an adverse event was recorded in the ED medical record or in the confidential hospital event reporting system. ED patients can identify adverse events affecting their care. Moreover, many of these events are not recorded in the medical record. Engaging patients and their family members in identification of errors may enhance patient safety.


Characterization of Antibodies that Detect Human GFAP after Traumatic Brain Injury.
After traumatic brain injury (TBI), glial fibrillary acidic protein (GFAP) and other brain-derived proteins and their breakdown products are released into biofluids such as CSF and blood. Recently, a sandwich ELISA was constructed that measured GFAP concentrations in CSF or serum from human mild-moderate TBI patients. The goals of the present study were to characterize the same two antibodies used in this ELISA, and to determine which GFAP bands are detected by this antibody combination. Here, both antibodies recognized GFAP specifically in human brain and post-TBI CSF in a cluster of bands ranging from 50-38 kDa, that resembled bands from calpain-cleaved GFAP. By immunoprecipitation, the anti-GFAP Capture antibody recovered full length GFAP and its breakdown products from human brain lysate and post-TBI CSF. These findings demonstrate that the anti-GFAP ELISA antibodies non-preferentially detect intact GFAP and GFAP breakdown products, underscoring their utility for detecting brain injury in human patients.


2-hydroxyoleic acid: a new hypotensive molecule.
Recent studies have shown that diets rich in monounsaturated fatty acids (MUFAs) from olive oil, a natural source of oleic acid, have beneficial effects on blood pressure (BP) in hypertensive patients. With this in mind, we investigated whether a synthetic derivative of the MUFA oleic acid, 2-hydroxyoleic acid (2-OHOA), was capable of regulating the BP of Sprague-Dawley rats. Intraperitoneal and oral administration of 2-OHOA to rats induced significant and sustained decreases in BP in a time-dependent manner. Without affecting heart rate, treatments for 7 days provoked reductions in systolic BP of 20 to 26 mm Hg. At the molecular level, the density of Galpha(s), but not Galpha(i2) or Galpha(o), increased in membranes from the hearts and aortas of 2-OHOA-treated rats, whereas in heart membranes, the density of Galpha(q)/11 and protein kinase Calpha proteins was also augmented. These molecular alterations were reflected in the increase in cAMP levels after Galpha(s) protein and beta-adrenergic receptor stimulation. On the contrary, inhibitory hormones reduced adenylyl cyclase activity to the same extent in 2-OHOA-treated rats as in vehicle-treated ones. Our results indicate that cardiovascular tissues from 2-OHOA-treated rats exhibited increased cAMP production in response to Galpha(s) activation, which might be attributed to enhanced expression of Galpha(s) proteins. As a result of this change, a significant reduction in systolic BP was observed. Therefore, BP can be lowered by administration of 2-OHOA, which might represent the first member of a new family of antihypertensive drugs.


[Epoxy embedding of bone-marrow biopsies for light microscopy and histopathologic use (author's transl)].
A routine method of epoxy embedding of bone-marrow and other tissues for light microscopy is described. Undecalcified bone specimens obtained from the sternum or iliac crest using trephine biopsy are fixed in formalin, deshydrated in acetone, embedded in a Kushida's low-viscosity epoxy resin medium. The samples are cut using a carbide-edged knife in a rotary microtome or a glass knife in a Sorval JB4 microtome. After fixing sections to slide with Neoprene and removal of the embedding medium with sodium methoxide and acetone, the staining with usual histological procedures is quite easy. A simple and rapid technique is outlined for staining these sections with Giemsa. The resolution, compared with routine parafin sections, is improved.


Vitamin and mineral supplements: do we really need them?
In the United States, 40 - 50% of the men and women 50 years of age or older regularly use multivitamin/mineral (MVM) supplements, making the annual sales of these supplements over $11 billion. However, the question remains whether using MVM supplements is beneficial to health. This article reviews the results of randomized studies of MVM supplements and individual vitamins/mineral supplements in relation to overall mortality and incidence of chronic diseases, particularly cancer and ischemic heart disease. The results of large-scale randomized trials show that, for the majority of the population, there is no overall benefit from taking MVM supplements. Indeed, some studies have shown increased risk of cancers in relation to using certain vitamins.


The environment topography alters the way to multicellularity in Myxococcus xanthus.
The social soil-dwelling bacterium Myxococcus xanthus can form multicellular structures, known as fruiting bodies. Experiments in homogeneous environments have shown that this process is affected by the physicochemical properties of the substrate, but they have largely neglected the role of complex topographies. We experimentally demonstrate that the topography alters single-cell motility and multicellular organization in M. xanthus In topographies realized by randomly placing silica particles over agar plates, we observe that the cells' interaction with particles drastically modifies the dynamics of cellular aggregation, leading to changes in the number, size, and shape of the fruiting bodies and even to arresting their formation in certain conditions. We further explore this type of cell-particle interaction in a computational model. These results provide fundamental insights into how the environment topography influences the emergence of complex multicellular structures from single cells, which is a fundamental problem of biological, ecological, and medical relevance.


Laparoscopic appendectomy trends and outcomes in the United States: data from the Nationwide Inpatient Sample (NIS), 2004-2011.
Laparoscopic appendectomy (LA) is becoming the standard procedure of choice for appendicitis. We aimed to evaluate the frequency and trends of LA for acute appendicitis in the United States and to compare outcomes of LA with open appendectomy (OA). Using the Nationwide Inpatient Sample database, we examined patients who underwent appendectomy for acute appendicitis from 2004 to 2011. A total of 2,593,786 patients underwent appendectomy during this period. Overall, the rate of LA was 60.5 per cent (children: 58.1%; adults: 63%; elderly: 48.7%). LA rate significantly increased from 43.3 per cent in 2004 to 75 per cent in 2011. LA use increased 66 per cent in nonperforated appendicitis versus 100 per cent increase in LA use for perforated appendicitis. The LA rate increased in all age groups. The increased LA use was more significant in male patients (84%) compared with female patients (62%). The overall conversion rate of LA to OA was 6.3 per cent. Compared with OA, LA had a significantly lower complication rate, a lower mortality rate, a shorter mean hospital stay, and lower mean total hospital charges in both nonperforated and perforated appendices. LA has become an established procedure for appendectomy in nonperforated and perforated appendicitis in all rates exceeding OA. Conversion rate is relatively low (6.3%).


Endotracheal tube-induced sore throat pain and inflammation is coupled to the release of mitochondrial DNA.
In the absence of infection, the pathophysiology of endotracheal tube-induced sore throat pain is unclear. Activated neutrophils release elastase, reactive oxygen species, and inflammatory cytokines known to contribute to neuropathic pain. Sterile tissue injury can cause the release of damage-associated molecular patterns such as mitochondrial DNA that promote neutrophil activation. We hypothesized that endotracheal tube-induced sore throat pain is linked to mitochondrial DNA-mediated neutrophil inflammation. A nonrandomized prospective survey for sore throat pain was conducted in 31 patients who required short-term intubation and had no evidence of upper airway infection. Patterns of neutrophil abundance, activation, and mitochondrial DNA levels were analyzed in tracheal lavage fluid following intubation and prior to extubation. Thirteen of 31 patients reported sore throat pain. Sore throat patients had high neutrophilia with elevated adhesion molecule and TLR9 expression and constitutive reactive oxygen species generation. Tracheal lavage fluid from sore throat patients accumulated mitochondrial DNA and stimulated neutrophils to release mediators associated with pain in a TLR9- and DNAse-dependent fashion. Endotracheal tube-induced sore throat is linked to the release of mitochondrial DNA and can drive TLR9-mediated inflammatory responses by neutrophils reported to cause pain. Mitigating the effects of cell-free mitochondrial DNA may prove beneficial for the prevention of endotracheal tube-mediated sore throat pain.


Effect of unilateral ibotenate lesions of the ventral tegmental area on cortical and hippocampal EEG in freely behaving rats.
It was found previously that unilateral destruction of the ventral tegmental area (VTA) facilitated behavioral responses (exploration, eating) induced by electrical stimulation of the contralateral VTA. The same effect occurred after unilateral injections of pharmacological agents, which led to a decrease in dopaminergic transmission in the VTA. While trying to explain the mechanism behind this "contralateral facilitation effect" in the present experiment we examined whether augmentation of function of the contralateral hemisphere would be reflected in cortical and hippocampal EEG changes in conscious rats. Unilateral, cytotoxic lesion of the VTA caused a bilateral decrease in neocortical and hippocampal EEG power during both exploratory sniffing and eating. Depression involved all the frequency bands in the prefrontal cortex, mainly in the hemisphere contralateral to the VTA lesion. In the hippocampus the depression was slightly more intense ipsilaterally, also involving all the frequency bands although to different degrees. The results indicate that the VTA is involved in the regulation of cortical and hippocampal activity during VTA-dependent behavioral activation, and that the "contralateral facilitation effect" is concomitant with lateralized changes in EEG activity.


Inhibition of Inflammatory Response by Crocin Attenuates Hemorrhagic Shock-Induced Organ Damages in Rats.
Hemorrhagic shock (HS) is associated with an excessive activation of inflammation, contributing to multiple organ failure in numerous medical or surgical conditions. To explore the therapeutic potential of crocin, a natural compound with anti-inflammatory properties, we administered crocin to rats during resuscitation following HS induced by withdrawing blood. Compared with control animals which were sham-treated, HS-operated rats showed organ damages as manifested by enhanced markers of multiple organ dysfunctions. Crocin treatment substantially reduced these parameters in rats subjected to HS, suggesting an alleviation of tissue injuries such as in the kidney, liver, pancreas, and muscle. The activation of NF-κB (nuclear factor κB) pathway in lung tissue by HS, as shown by increased nuclear translocation of p65 and IκBα phosphorylation, was diminished by crocin treatment. The crocin administration also significantly decreased the serum levels of proinflammatory cytokine TNF-α (tumor necrosis factor-α) and interleukin (IL)-6, whereas increased the level of anti-inflammatory cytokine IL-10 in HS-operated rats. These studies indicate that crocin administration may reduce inflammation-driven tissue damage in patients with HS.


Median nerve entrapment in a pediatric both-bone forearm fracture: recognition and management in the acute setting.
Entrapment of the median nerve in pediatric both-bone forearm fractures is a rare occurrence, but there are multiple reports of such cases in the literature. This case report discusses the recognition and management of median nerve entrapment in a 13-year-old female who presented acutely with a both-bone forearm fracture and a reproducible neurologic deficit in the median nerve distribution. She was treated with acute open reduction and internal plate fixation and median nerve exploration. Her median nerve function was near normal 14 weeks after surgery. This report stresses the importance of a complete neurologic examination in all forearm fractures and demonstrates the recovery potential of a median nerve damaged by bony entrapment.


Functional roles of the A335 and G338 residues of the proton-coupled folate transporter (PCFT-SLC46A1) mutated in hereditary folate malabsorption.
The proton-coupled folate transporter (PCFT-SLC46A1) mediates intestinal folate absorption and folate transport across the choroid plexus, processes defective in hereditary folate malabsorption (HFM). This paper characterizes the functional defect, and the roles of two mutated PCFT residues, associated with HFM (G338R and A335D). The A335D-PCFT and other mutations at this residue result in an unstable protein; when expression of a mutant protein was preserved, function was always retained. The G338R and other charged mutants resulted in an unstable protein; substitutions with small neutral and polar amino acids preserved protein but with impaired function. Pemetrexed and methotrexate (MTX) influx kinetics mediated by the G338C mutant PCFT revealed marked (15- to 20-fold) decreases in K(t) and V(max) compared with wild-type PCFT. In contrast, there was only a small (∼2-fold) decrease in the MTX influx K(i) and an increase (∼3-fold) in the pemetrexed influx K(i) for the G338C-PCFT mutant. Neither a decrease in pH to 4.5, nor an increase to 7.4, restored function of the G338C mutant relative to wild-type PCFT excluding a role for this residue in proton binding or proton coupling. Homology modeling localized the A335 and G338 residues embedded in the 9th transmembrane, consistent with the inaccessibility of the A335C and G338C proteins to MTS reagents. Hence, the loss of intrinsic G338C-PCFT function was due solely to impaired oscillation of the carrier between its conformational states. The data illustrate how alterations in carrier cycling can impact influx K(t) without comparable alterations in substrate binding to the carrier.


Robust anisotropic Gaussian fitting for volumetric characterization of pulmonary nodules in multislice CT.
This paper proposes a robust statistical estimation and verification framework for characterizing the ellipsoidal (anisotropic) geometrical structure of pulmonary nodules in the Multislice X-ray computed tomography (CT) images. Given a marker indicating a rough location of a target, the proposed solution estimates the target's center location, ellipsoidal boundary approximation, volume, maximum/average diameters, and isotropy by robustly and efficiently fitting an anisotropic Gaussian intensity model. We propose a novel multiscale joint segmentation and model fitting solution which extends the robust mean shift-based analysis to the linear scale-space theory. The design is motivated for enhancing the robustness against margin-truncation induced by neighboring structures, data with large deviations from the chosen model, and marker location variability. A chi-square-based statistical verification and analytical volumetric measurement solutions are also proposed to complement this estimation framework. Experiments with synthetic one-dimensional and two-dimensional data clearly demonstrate the advantage of our solution in comparison with the gamma-normalized Laplacian approach (Linderberg, 1998) and the standard sample estimation approach (Matei, 2001). A quasi-real-time three-dimensional nodule characterization system is developed using this framework and validated with two clinical data sets of thin-section chest CT images. Our experiments with 1310 nodules resulted in (1) robustness against intraoperator and interoperator variability due to varying marker locations, (2) 81% correct estimation rate, (3) 3% false acceptance and 5% false rejection rates, and (4) correct characterization of clinically significant nonsolid ground-glass opacity nodules. This system processes each 33-voxel volume-of-interest by an average of 2 s with a 2.4-GHz Intel CPU. Our solution is generic and can be applied for the analysis of blob-like structures in various other applications.


Impact of 2'-hydroxyl sampling on the conformational properties of RNA: update of the CHARMM all-atom additive force field for RNA.
Here, we present an update of the CHARMM27 all-atom additive force field for nucleic acids that improves the treatment of RNA molecules. The original CHARMM27 force field parameters exhibit enhanced Watson-Crick base pair opening which is not consistent with experiment, whereas analysis of molecular dynamics (MD) simulations show the 2'-hydroxyl moiety to almost exclusively sample the O3' orientation. Quantum mechanical (QM) studies of RNA related model compounds indicate the energy minimum associated with the O3' orientation to be too favorable, consistent with the MD results. Optimization of the dihedral parameters dictating the energy of the 2'-hydroxyl proton targeting the QM data yielded several parameter sets, which sample both the base and O3' orientations of the 2'-hydroxyl to varying degrees. Selection of the final dihedral parameters was based on reproduction of hydration behavior as related to a survey of crystallographic data and better agreement with experimental NMR J-coupling values. Application of the model, designated CHARMM36, to a collection of canonical and noncanonical RNA molecules reveals overall improved agreement with a range of experimental observables as compared to CHARMM27. The results also indicate the sensitivity of the conformational heterogeneity of RNA to the orientation of the 2'-hydroxyl moiety and support a model whereby the 2'-hydroxyl can enhance the probability of conformational transitions in RNA.


Endoscopic management of pain in pancreatic cancer.
Pancreatic cancer is the fourth leading cause of cancer-related death in the United States and one of the leading causes of cancer mortality in the United States. Due to its aggressive behavior and lack of effective therapies, palliation plays a critical role in the management of the disease. Most patients with pancreatic cancer suffer from severe pain, which adversely predicts prognosis and significantly impacts the quality of life. Therefore pain management plays a central role in palliation. Non-steroidal anti-inflammatory drugs and opioid agents are often first line medications in pain management, but they do not target the underlying pathophysiology of pain and their use is limited by adverse effects and dependence. The proposed mechanisms of pain development in pancreatic cancer include neurogenic inflammation and ductal hypertension which may be targeted by endoscopic therapies. Endoscopic ultrasound-guided celiac plexus neurolysis (EUS-CPN) and pancreatic duct stent placement are the two primary endoscopic modalities for palliative management in pancreatic cancer patients with refractory pain. Other endoscopic treatments such as biliary stent placement and enteral stent placement for biliary and duodenal obstruction may also help palliate pain in addition to their role in decompression. This article reviews the existing evidence for these endoscopic interventions for pain management in pancreatic cancer.


Crystal structures and enzyme mechanisms of a dual fucose mutarotase/ribose pyranase.
Escherichia coli FucU (Fucose Unknown) is a dual fucose mutarotase and ribose pyranase, which shares 44% sequence identity with its human counterpart. Herein, we report the structures of E. coli FucU and mouse FucU bound to L-fucose and delineate the catalytic mechanisms underlying the interconversion between stereoisomers of fucose and ribose. E. coli FucU forms a decameric toroid with each active site formed by two adjacent subunits. While one subunit provides most of the fucose-interacting residues including a catalytic tyrosine residue, the other subunit provides a catalytic His-Asp dyad. This active-site feature is critical not only for the mutarotase activity toward L-fucose but also for the pyranase activity toward D-ribose. Structural and biochemical analyses pointed that mouse FucU assembles into four different oligomeric forms, among which the smallest homodimeric form is most abundant and would be the predominant species under physiological conditions. This homodimer has two fucose-binding sites that are devoid of the His-Asp dyad and catalytically inactive, indicating that the mutarotase and the pyranase activities appear dispensable in vertebrates. The defective assembly of the mouse FucU homodimer into the decameric form is due to an insertion of two residues at the N-terminal extreme, which is a common aspect of all the known vertebrate FucU proteins. Therefore, vertebrate FucU appears to serve for as yet unknown function through the quaternary structural alteration.


A diffuse lung emphysema, severe pulmonary hypertension and lack of airflow limitation.
Pulmonary veno-occlusive disease is characterized by remodeling of pulmonary arteries, capillaries and venules. We report a case of diffuse lung emphysema and pulmonary veno-occlusive disease with the characteristic of having no airflow limitation. A very low diffusing capacity for carbon monoxide and results of high-resolution computed tomography of the chest suggested pulmonary veno-occlusive disease. The diagnosis was confirmed on histological analysis after lung transplantation. The combination of results of the computed tomography of the chest and the histological analysis suggested a relationship between diffuse lung emphysema and remodeling of pulmonary vessels. A distinctive pattern of mild-to-moderate airflow limitation in patients with chronic obstructive pulmonary disease and severe pulmonary hypertension has been described. This observation of the combination of diffuse emphysema, pulmonary veno-occlusive disease and no airflow limitation supports further pathophysiological studies on severe pulmonary hypertension in chronic obstructive pulmonary disease.


Resveratrol suppresses IGF-1 induced human colon cancer cell proliferation and elevates apoptosis via suppression of IGF-1R/Wnt and activation of p53 signaling pathways.
Obesity is a global phenomenon and is associated with various types of cancer, including colon cancer. There is a growing interest for safe and effective bioactive compounds that suppress the risk for obesity-promoted colon cancer. Resveratrol (trans-3, 4', 5,-trihydroxystilbene), a stilbenoid found in the skin of red grapes and peanuts suppresses many types of cancers by regulating cell proliferation and apoptosis through a variety of mechanisms, however, resveratrol effects on obesity-promoted colon cancer are not clearly established. We investigated the anti-proliferative effects of resveratrol on HT-29 and SW480 human colon cancer cells in the presence and absence of insulin like growth factor-1 (IGF-1; elevated during obesity) and elucidated the mechanisms of action using IGF-1R siRNA in HT-29 cells which represents advanced colon carcinogenesis. Resveratrol (100-150 microM) exhibited anti-proliferative properties in HT-29 cells even after IGF-1 exposure by arresting G0/G1-S phase cell cycle progression through p27 stimulation and cyclin D1 suppression. Treatment with resveratrol suppressed IGF-1R protein levels and concurrently attenuated the downstream Akt/Wnt signaling pathways that play a critical role in cell proliferation. Targeted suppression of IGF-1R using IGF-1R siRNA also affected these signaling pathways in a similar manner. Resveratrol treatment induced apoptosis by activating tumor suppressor p53 protein, whereas IGF-1R siRNA treatment did not affect apoptosis. Our data suggests that resveratrol not only suppresses cell proliferation by inhibiting IGF-1R and its downstream signaling pathways similar to that of IGF-1R siRNA but also enhances apoptosis via activation of the p53 pathway. For the first time, we report that resveratrol suppresses colon cancer cell proliferation and elevates apoptosis even in the presence of IGF-1 via suppression of IGF-1R/Akt/Wnt signaling pathways and activation of p53, suggesting its potential role as a chemotherapeutic agent.


Thermal effects on sudden changes and freezing of correlations between remote atoms in a cavity quantum electrodynamics network.
We investigate thermal effects on sudden changes and freezing of the quantum and classical correlations of remote qubits in a cavity quantum electrodynamics (CQED) network with losses. We find that the detrimental effect of thermal reservoirs on the freezing of correlations can be compensated via an efficient coupling of the fiber connecting the two cavities of the system. Furthermore, for certain initial conditions, we find a double sudden transition in the dynamics of Bures geometrical quantum discord. The second transition tends to disappear at a critical temperature, hence freezing the discord. Finally, we discuss the feasibility of the experimental realization of the present proposal.


High prevalence of orofacial clefts in Shanxi Province in northern China, 2003-2004.
Shanxi Province in northern China has been identified as an area with the highest prevalence of neural tube defects in the world; however, few reports exist on the prevalence of orofacial clefts (OFC). We examined the prevalence of OFC and their association with selected characteristics. Data came from a population-based birth defects surveillance system in four counties of Shanxi Province. The system captures information on all live births, stillbirths of at least 20 weeks gestation, and pregnancy terminations at any gestational age after prenatal diagnosis of a birth defect. Out of 25,355 births in 2003-2004, 83 cases with orofacial clefts were identified resulting in a birth prevalence of 3.27 per 1,000. Other major external birth defects were present in 12 (14.5%) OFC cases. Cleft lip with palate (CLP), cleft lip alone (CL) and cleft palate alone (CP) accounted for 62.0%, 29.6% and 8.4% of OFC cases without additional external defects, respectively. The male to female ratio was 1.04 (95%CI: 0.67-1.60) for all OFC, 1.46 (95%CI: 0.80-2.68) for CLP, 1.23 (95%CI: 0.52-2.91) for CL, 0.46 (95%CI: 0.08-2.51) for CP without additional major external defects, and 0.31(95%CI: 0.08-1.13) for OFC with additional external defects. The prevalence of OFC without additional external defects was increased with decreased maternal age (P < 0.05) and increased maternal education (P < 0.05). The birth prevalence of OFC in Shanxi Province of northern China is among the highest worldwide. Further studies are required to define the factors associated with excess risk of clefts.


Sputum long non-coding RNA biomarkers for diagnosis of lung cancer.
Analysis of molecular changes in sputum may help diagnose lung cancer. Long non-coding RNAs (lncRNAs) play vital roles in various biological processes, and their dysregulations contribute to the development and progression of lung tumorigenesis. Herein, we determine whether aberrant lncRNAs could be used as potential sputum biomarkers for lung cancer. Using reverse transcription PCR, we measure expressions of lung cancer-associated lncRNAs in sputum of a discovery cohort of 67 lung cancer patients and 65 cancer-free smokers with benign diseases and a validation cohort of 59 lung cancer patients and 60 cancer-free smokers with benign diseases. In the discovery cohort, four of the lncRNAs displayed a significantly different level in sputum of lung cancer patients vs.cancer-free smokers with benign diseases (all P< 0.001). From the four lncRNAs, three lncRNAs (SNHG1, H19, and HOTAIR) are identified as a biomarker panel, producing 82.09% sensitivity and 89.23% specificity for diagnosis of lung cancer. Furthermore, the biomarker panel has a higher sensitivity (82.09% vs. 52.24%, P= 0.02) and a similar specificity compared with sputum cytology (89.23% vs. 90.77%, P= 0.45). In addition, the lncRNA biomarker panel had a higher sensitivity (87.50% vs. 70.07%, p= 0.03) for diagnosis of squamous cell carcinoma compared with adenocarcinoma of the lung, while maintaining the same specificity (89.23%). The potential of the sputum lncRNA biomarkers for lung cancer detection is confirmed in the validation cohort. We have for the first time shown that the analysis of lncRNAs in sputum might be a noninvasive approach for diagnosis of lung cancer.


Improvement of ablative margins by the intraoperative use of CEUS-CT/MR image fusion in hepatocellular carcinoma.
To assess whether intraoperative use of contrast-enhanced ultrasound (CEUS)-CT/MR image fusion can accurately evaluate ablative margin (AM) and guide supplementary ablation to improve AM after hepatocellular carcinoma (HCC) ablation. Ninety-eight patients with 126 HCCs designated to undergo thermal ablation treatment were enrolled in this prospective study. CEUS-CT/MR image fusion was performed intraoperatively to evaluate whether 5-mm AM was covered by the ablative area. If possible, supplementary ablation was applied at the site of inadequate AM. The CEUS image quality, the time used for CEUS-CT/MR image fusion and the success rate of image fusion were recorded. Local tumor progression (LTP) was observed during follow-up. Clinical factors including AM were examined to identify risk factors for LTP. The success rate of image fusion was 96.2% (126/131), and the duration required for image fusion was 4.9 ± 2.0 (3-13) min. The CEUS image quality was good in 36.1% (53/147) and medium in 63.9% (94/147) of the cases. By supplementary ablation, 21.8% (12/55) of lesions with inadequate AMs became adequate AMs. During follow-up, there were 5 LTPs in lesions with inadequate AMs and 1 LTP in lesions with adequate AMs. Multivariate analysis showed that AM was the only independent risk factor for LTP (hazard ratio, 9.167; 95% confidence interval, 1.070-78.571; p = 0.043). CEUS-CT/MR image fusion is feasible for intraoperative use and can serve as an accurate method to evaluate AMs and guide supplementary ablation to lower inadequate AMs.


The use of imatinib mesylate has no adverse effects on the heart function. Results of a pilot study in patients with chronic myeloid leukemia.
To investigate cardiac effects of imatinib at an extended follow-up (median 12.4 months), 12 chronic myeloid leukemia patients underwent cardiac screening. No significant changes on the frequency of cardiovascular signs and symptoms, electrocardiographic abnormalities, echocardiographic measurements and BNP levels were observed. Median ejection fraction was 67% at baseline versus 68% at follow-up (median intra-patient change 0.5%). Median BNP levels were 8.3 versus 7.3pg/mL (median intra-patient change 0.2pg/mL). Troponin I measures were below the lower limit of detection, whereas strain measures were similar to healthy control. This pilot study suggests that it is probably safe to perform cardiac monitoring on an annual basis.


Crude drugs as anticancer agents.
Although tremendous progress has been made in basic cancer biology and in the development of novel cancer treatments, cancer remains a leading cause of death in the world. The etiopathogenesis of cancer is complex. Besides genetic predisposition, known environmental factors associated with cancer are: diet, lifestyle, and environmental toxins. Toxicity of drugs and eventual relapse of cancers contribute to high cancer death rates. Current therapeutic interventions for cancer- surgery, chemotherapy, radiotherapy, thermotherapy, etc. are far from being curative for many forms of cancer. Chemotherapy, in particular, though the most commonly used cancer treatment, is usually associated with side effects with varying degrees of severity. The purpose of this brief review is to assemble current literature on some crude drugs and to focus on their beneficial roles and drug targets in cancer therapy and chemo-prevention. Although their pharmacological mechanisms and biochemical roles in cancer biology and tumor chemo-prevention are not fully understood, crude drugs are believed to have nutriceutical effects upon cancer patients.


Detection of hemodynamically significant coronary artery stenosis: incremental diagnostic value of dynamic CT-based myocardial perfusion imaging.
To determine the feasibility of computed tomography (CT)-based dynamic myocardial perfusion imaging for the detection of hemodynamically significant coronary artery stenosis, as defined with fractional flow reserve (FFR). Institutional review board approval and informed patient consent were obtained before patient enrollment in the study. The study was HIPAA compliant. Subjects who were suspected of having or were known to have coronary artery disease underwent electrocardiographically triggered dynamic stress myocardial perfusion imaging. FFR measurement was performed within all main coronary arteries with a luminal narrowing of 50%-85%. Estimated myocardial blood flow (MBF) was derived from CT images by using a model-based parametric deconvolution method for 16 myocardial segments and was related to hemodynamically significant coronary artery stenosis with an FFR of 0.75 or less in a blinded fashion. Conventional measures of diagnostic accuracy were derived, and discriminatory power analysis was performed by using logistic regression analysis. Of 36 enrolled subjects, 33 (mean age, 68.1 years ± 10 [standard deviation]; 25 [76%] men, eight [24%] women) completed the study protocol. An MBF cut point of 75 mL/100 mL/min provided the highest discriminatory power (C statistic, 0.707; P <.001). While the diagnostic accuracy of CT for the detection of anatomically significant coronary artery stenosis (>50%) was high, it was low for the detection of hemodynamically significant stenosis (positive predictive value [PPV] per coronary segment, 49%; 95% confidence interval [CI]: 36%, 60%). With use of estimated MBF to reclassify lesions depicted with CT angiography, 30 of 70 (43%) coronary lesions were graded as not hemodynamically significant, which significantly increased PPV to 78% (95% CI: 61%, 89%; P = .02). The presence of a coronary artery stenosis with a corresponding MBF less than 75 mL/100 mL/min had a high risk for hemodynamic significance (odds ratio, 86.9; 95% CI:17.6, 430.4). Dynamic CT-based stress myocardial perfusion imaging may allow detection of hemodynamically significant coronary artery stenosis.


Comparison of antimicrobial susceptibility among Clostridium difficile isolated from an integrated human and swine population in Texas.
Clostridium difficile can be a major problem in hospitals because the bacterium primarily affects individuals with an altered intestinal flora; this largely occurs through prolonged antibiotic use. Proposed sources of increased community-acquired infections are food animals and retail meats. The objective of this study was to compare the antimicrobial resistance patterns of C. difficile isolated from a closed, integrated population of humans and swine to increase understanding of the bacterium in these populations. Swine fecal samples were collected from a vertically flowing swine population consisting of farrowing, nursery, breeding, and grower/finisher production groups. Human wastewater samples were collected from swine worker and nonworker occupational group cohorts. Antimicrobial susceptibility testing was performed on 523 C. difficile strains from the population using the commercially available agar diffusion Epsilometer test (Etest(®)) for 11 different antimicrobials. All of the swine and human strains were susceptible to amoxicillin/clavulanic acid, piperacillin/tazobactam, and vancomycin. In addition, all of the human strains were susceptible to chloramphenicol. The majority of the human and swine strains were resistant to cefoxitin and ciprofloxacin. Statistically significant differences in antimicrobial susceptibility were found among the swine production groups for ciprofloxacin, tetracycline, amoxicillin/clavulanic acid, and clindamycin. No significant differences in antimicrobial susceptibility were found across human occupational group cohorts. We found that 8.3% of the swine strains and 13.3% of the human strains exhibited resistance to metronidazole. The finding of differences in susceptibility patterns between human and swine strains of C. difficile provides evidence that transmission between host species in this integrated population is unlikely.


Enhancement of 2-photon absorption of a dye in a polymer microsphere based on an optical cavity effect.
We examined optical cavity effects on the 2-photon absorption efficiency of a dye (Rohdamine B; RhB) doped in polymer microspheres (ion-exchanging resin and poly(methyl methacrylate) (PMMA)). When 1064 nm pulsed-laser light was irradiated on single polymer microspheres doped with RhB under an optical microscope, we clearly observed dye fluorescence from individual microspheres, although the dye was transparent at 1064 nm. We confirmed that the fluorescence intensity was proportional to the square of the laser power for excitation. In contrast, such fluorescence was not observed from RhB doped in a PMMA film, in which the enhancement of a light electric field by an optical cavity effect was never expected. Theoretical calculations indicated that the microsphere possessed several peak values of the quality factor (Q = 10(2)-10(5)) at around certain particle diameters under 1064 nm irradiation. This means that the electromagnetic field of incident light is enhanced through light confinement in a microsphere. Based on these results, we conclude that the 2-photon absorption probability of RhB would be considerably enhanced by the optical cavity effects of the microsphere.


Metabolism of human femoral head cartilage in osteoarthrosis and subcapital fracture.
The cell density and incorporation of 35SO4 and 3H-glycine into human articular cartilage from 8 osteoarthrotic and 7 normal (subcapital fracture) femoral heads were studied. It was found that osteoarthrotic cartilage incorporates on a per cell basis about twice as much 35SO4 and 2--5 times as much 3H-glycine as normal cartilage. There was no relationship between the intensity of incorporation and either the location of the cartilage (weight-bearing versus non weight-bearing areas) in normal cartilage or the degree of damage (normal-like, fibrillated, and ulcerated) in osteoarthrotic articular cartilage. In the latter tissue the increased synthetic capacity of the cells seems to be a diffuse rather than a localised process, for it was also found in cartilage from peripheral osteophytes. Histo-autoradiographic studies showed that the osteoarthrotic chondrocytes are metabolically hyperactive all over the femoral head, including wedge-shaped margins of the zone of exposed bone. These results support the hypothesis that much of the articular cartilage from osteoarthrotic femoral heads is of an immature chondroblastic type. It is suggested that de-novo synthesis of articular cartilage occurs during the process of regional remodelling of the femoral head, which would account for the observed hyperactivity.


In-Source CID Ramping and Covariant Ion Analysis of Hydrophilic Interaction Chromatography Metabolomics.
A large proportion of the complexity and redundancy of LC-MS metabolomics data comes from adduct formation. To reduce such redundancy, many tools have been developed to recognize and annotate adduct ions. These tools rely on predefined adduct lists that are generated empirically from reversed-phase LC-MS studies. In addition, hydrophilic interaction chromatography (HILIC) is gaining popularity in metabolomics studies due to its enhanced performance over other methods for polar compounds. HILIC methods typically use high concentrations of buffer salts to improve chromatographic performance. Therefore, it is necessary to analyze adduct formation in HILIC metabolomics. To this end, we developed covariant ion analysis (COVINA) to investigate metabolite adduct formation. Using this tool, we completely annotated 201 adduct and fragment ions from 10 metabolites. Many of the metabolite adduct ions were found to contain cluster ions corresponding to mobile phase additives. We further utilized COVINA to find the major ionized forms of metabolites. Our results show that for some metabolites, the adduct ion signals can be >200-fold higher than the signals from the deprotonated form, offering better sensitivity for targeted metabolomics analysis. Finally, we developed an in-source CID ramping (InCIDR) method to analyze the intensity changes of the adduct and fragment ions from metabolites. Our analysis demonstrates a promising method to distinguish the protonated and deprotonated ions of metabolites from the adduct and fragment ions.


Yeast species isolated from Texas High Plains vineyards and dynamics during spontaneous fermentations of Tempranillo grapes.
Vineyards and grape musts harbor complex locally specific microbial communities, among which yeast species can be responsible of spontaneous alcoholic fermentation. Although relying on indigenous yeast can be a risk for winemaking, local yeast diversity is associated with complexity and stronger identity of the wine produced, compared to inoculated alcoholic fermentation with commercial yeast strains. In this context, the main yeast species present on grapes, leaves and soils of Tempranillo and Cabernet Sauvignon vineyards in the hot semi-arid climate of the Texas High Plains area were investigated, as well as the presence and dynamics of yeast species during spontaneous fermentations of Tempranillo grapes from the same vineyards. Molecular characterization of yeast species was performed using culture-dependent 5.8S-ITS restriction fragment length polymorphism method and sequencing. Yeast species recovered from grapes, leaves, and soils were mainly dominated by Aureobasidium pullulans, Cryptococcus species, Filobasidium species and Naganishia species, typical members of the vineyard environment. One isolate of potential enological interest, Lachancea thermotolerans, a fermenting yeast with potential in must acidification, was recovered from the vineyard environment. However, spontaneous alcoholic fermentations revealed the presence of fermenting yeast species, including Saccharomyces cerevisiae, Lachancea thermotolerans and Hanseniaspora species. The presence of the three aforementioned species is of extreme interest for winemaking in the Texas High Plains area. Indeed, Saccharomyces cerevisiae is the model for alcoholic fermentation, Hanseniaspora species have been shown to improve palatability of wines, and Lachancea thermotolerans has become of increasing interest due to its potential to acidify musts and palatability. One of the main characteristics of grapes grown in the Texas High Plains area being the lack of acidity, focusing on these three yeast species could promote the development of locally oriented started cultures for the production of wines with a stronger local identity.


[Ultrastructural analysis of the effects of dominant cataract-Fr gene in mouse embryos].
An electron microscope study of lenses in 11--15 and 17 days old embryos of mice homozygous by dominant cataract-Fr (CatFr) gene has shown that ultrastructural changes in the nuclear envelope are the earliest expression of the mutant gene. The primary lens fibers of 12 days old embryos CatFr/CatFr, unlike those of the normal ones, are characterized by the decrease in the number of nuclear pores, evagination of the outer nuclear membrane, marked and unequal extension of perinuclear space which connects itself with the endoplasmic reticulum cisternae. In 14 days old embryos breaks in the outer nuclear membrane and evaginations in the inner one, fusion of nuclear membranes and breaks of the nuclear envelope are observed and resulted in the release of the nuclear contents with the nucleolus in the cytoplasm. Similar ultrastructural changes are characteristic also of the nuclei of secondary lens fibers at a comparable stage of differentiation. The destructive changes of nuclei are accompanied by the degeneration and autophagocytosis of cellular organelles and matrix.


Relation between clinical, angiographic and ischemic findings at baseline and ischemia-related adverse outcomes at 1 year in the Asymptomatic Cardiac Ischemia Pilot study. ACIP Study Group.
We attempted to investigate the relation between patient characteristics and adverse outcome in patients with ischemia and clinically stable coronary artery disease (CAD). Evidence suggests that cardiac ischemia, detected by exercise stress testing (ETT) and ambulatory electrocardiographic (AECG) monitoring during daily living, identifies a subgroup of patients at increased risk for adverse outcome, but the relation between these ischemia findings and clinical and angiographic characteristics is largely unknown. We examined the relation between clinical, angiographic and ischemia characteristics at entry with adverse outcome observed at 1 year in the 558 patients enrolled in the Asymptomatic Cardiac Ischemia Pilot (ACIP) study. By the 12-month visit 13.1% of patients had an ischemia-related adverse clinical outcome that included death, nonfatal myocardial infarction or an ischemia-related hospital admission. Multivariate analysis identified only the number of AECG ischemic episodes at entry (odds ratio [OR] 1.06, 99% confidence interval [CI] 1.01 to 1.12, p = 0.002) as an independent predictor of outcome. Assignment to revascularization (as opposed to an initial medical treatment strategy) showed a trend (OR 0.56, 99% CI 0.26 to 1.2, p = 0.05). None of the other baseline clinical, exercise or angiographic variables examined provided additional information relative to adverse outcome. Determinants of adverse outcome, among clinically stable patients with CAD and ischemia induced by stress and daily life were magnitude of AECG ischemia before treatment and, possibly, initial treatment assignment. Among the many other characteristics examined, including age, symptom status and angiographic and exercise variables, none contributed additional independent prognostic information. These two simple variables, which may be modifiable, need further study in a larger trial.


Routine ultrasound examination during pregnancy: a world of possibilities.
to identify and describe the meaning of the routine ultrasound scan to pregnant women. a qualitative descriptive study using a grounded theory approach, with individual interviews to collect data. three antenatal clinics in a Swedish county of approximately 400,000 inhabitants. voluntary samples of 10 pregnant Swedish women, 26-38 years of age, were interviewed prior to their first routine ultrasound. 'making it possible' was the core category that explained and illustrated the meaning of the scan. The core category showed that the women considered the examination to be filled with possibilities to reach different goals during pregnancy. It also explained the categories: ultrasound as an event; ultrasound as a situation; ultrasound as a test; and the effects of ultrasound; as well as how they related to each other. The findings are considered the beginning of a theory concerning the meaning of the first ultrasound to pregnant women. pregnant women can see their first ultrasound as a tool that enables them to reach different goals during their pregnancy. Many of the goals concern meeting and connecting with the baby, suggesting that pregnant women consider the examination an important step towards parenthood. An ultrasound examination offered for medical reasons, which has other meanings than the intended for pregnant women, is important knowledge. It can be useful when giving information about the scan, addressing the woman during the examination, and for understanding and handling possible reactions.


[Modifying effect of low-molecular metabolities on the state of extracellular matrix of connective tissue of animals].
On the basis of complex approach to bone and haematopoetic tissue interaction the authors studied the influence of low weight metabolites on stromal fibroblasts and components of extacellular matrix of bone and skin (collagen and glycosaminoglycans). Specificity of different metabolites action on physico-chemical abilities of type I collagen, amino acid composition changes, surface charge, ratio of alpha- and beta-compounds, BrCN-fragments of alpha-1 component cross-links was shown. The dose dependence of formiate effect on processes of proteins glycosilation, cross-linking in bone and cartilage connective tissue and serum glycoproteins was established. The results obtained showed sensitivity of the bone tissue exstacellular matrix to influence of low-weight metabolites action at the level of post-synthetic modification of its components, their intermolecular interaction and process of osteogenesis.


Amiodarone-Induced Thyroid Storm Causing Sustained Monomorphic Ventricular Tachycardia Treated With Plasmapheresis: A Challenging Clinical Case.
Thyroid storm is a rare endocrine emergency with a high mortality rate approaching 20%. It manifests with the exaggerated symptoms of hyperthyroidism including hyperpyrexia, hypotension, cardiac arrhythmias, and death. The mainstay of treatment includes supportive intensive care and initiation of thionamides, beta blockers, corticosteroids, and if necessary anti-arrhythmics in the presence of refractory ventricular arrhythmias. We describe a case of amiodarone-induced thyrotoxicosis and thyroid storm leading to refractory ventricular tachycardias treated with plasmapheresis when anti-arrhythmic therapy became contraindicated. Amiodarone-induced thyroid storm can lead to monomorphic ventricular tachycardia. Under the circumstances of limited medical therapies, plasmapheresis can be an effective treatment option.


Arterial ischemic stroke (AIS) in childhood: clinical report from a single control center.
Stroke is the clinical designation for a rapidly developing loss of brain function due to an interruption in the blood supply to all or part of the brain. It is the third cause of death in adults and one of the top 10 causes in pediatric age. The perinatal period of onset is the second only to adult age group in the incidence of stroke. Arterial ischemic stroke during childhood occurs most frequently in the perinatal period with an incidence of 1 out 2300-5000 live infant births. This is a retrospective study that includes 28 patients affected by perinatal arterial ischemic stroke. Family and gestational history, risk factors of perinatal stroke, gender and clinical data of affected children and outcome are reported. A stroke family history was registered in three unrelated families. Gestational history disclosed cases of threats of abortion, preterm delivery, hyperthermia, gestosis, and placental disorders. In the children, onset of seizures were reported within 3 days of life and diagnosis of stroke was confirmed by brain MRI which disclosed involvement of the middle cerebral artery in all the cases. Hemilateral cerebral palsy, epileptic seizures, and intellectual disability from mild to severe were the most frequent complications. Stroke is still a common and dreadful events in perinatal period as this disorder is often unpredictable and cause of severe neurological impairment.


Enhance ZnO Photocatalytic Performance via Radiation Modified g-C3N4.
Environmental pollution, especially water pollution, is becoming increasingly serious. Organic dyes are one type of the harmful pollutants that pollute groundwater and destroy ecosystems. In this work, a series of graphitic carbon nitride (g-C3N4)/ZnO photocatalysts were facilely synthesized through a grinding method using ZnO nanoparticles and g-C3N4 as the starting materials. According to the results, the photocatalytic performance of 10 wt.% CN-200/Z-500 (CN-200, which g-C3N4 was 200 kGy, referred to the irradiation metering. Z-500, which ZnO was 500 °C, referred to the calcination temperature) with the CN-200 exposed to electron beam radiation was better than those of either Z-500 or CN-200 alone. This material displayed a 98.9% degradation rate of MB (20 mg/L) in 120 min. The improvement of the photocatalytic performance of the 10 wt.% CN-200/Z-500 composite material was caused by the improvement of the separation efficiency of photoinduced electron-hole pairs, which was, in turn, due to the formation of heterojunctions between CN-200 and Z-500 interfaces. Thus, this study proposes the application of electron-beam irradiation technology for the modification of photocatalytic materials and the improvement of photocatalytic performance.


Cardiac Tamponade after Thrombolysis for Acute Ischemic Stroke A Case with an Excellent Outcome.
Intravenous thrombolysis with recombinant tissue plasminogen activator (rtPA) is the established treatment for acute ischemic stroke and has been highly effective in reducing the neurological deficit. Serious adverse events are not uncommon, with hemorrhage being the major complication. We describe the case of a patient with acute ischemic stroke that also presented with vague cardiac symptoms and was treated with rtPA, which was complicated by a hemopericardium causing cardiac tamponade. Pericardiocentesis was promptly performed, which resulted in rapid resolution of the cardiogenic shock. The patient recovered consciousness within a few minutes. A search of the MEDLINE database shows that this is the first report of cardiac tamponade after rtPA thrombolysis occurring in a patient with no history of recent myocardial infarction or aortic dissection. Cardiac tamponade is a medical emergency and if not recognized and treated quickly results in cardiogenic shock and death. Hence, it is necessary to be aware that the established treatment for acute ischemic stroke can lead to cardiac tamponade.Cardiac tamponade after rtPA thrombolysis can occur in patients with no history of recent myocardial infarction or aortic dissection.


Bayesian estimation of weak material dispersion: theory and experiment.
This work considers the estimation of dispersion in materials via an interferometric technique. At its core, the problem involves extracting the quadratic variation in phase over a range of wavelengths based on measured optical intensity. The estimation problem becomes extremely difficult for weakly dispersive materials where the quadratic nonlinearity is very small relative to the uncertainty inherent in experiment. This work provides a means of estimating dispersion in the face of such uncertainty. Specifically, we use a Markov Chain Monte Carlo implementation of Bayesian analysis to provide both the dispersion estimate and the associated confidence interval. The interplay between various system parameters and the size of the resulting confidence interval is discussed. The approach is then applied to several different experimental samples.


Citrinin as a possible cause of the pruritus, pyrexia, haemorrhagic syndrome in cattle.
An outbreak of the pruritus, pyrexia, haemorrhagic syndrome affected eight of a herd of 175 cows which was divided into two groups of 115 and 60 according to yield. There was no difference in management between them but citrus pulp pellets were fed only to the larger group in which the eight cows were affected. Silage, which had been made without the use of additives, was also fed to both groups. The citrus pulp was visibly mouldy and contained 30 to 40 parts per billion of citrinin. Signs of the syndrome occurred within three days of the cows starting to ingest the citrus pulp, which was fed for 21 days, and the last case occurred six days after the feeding of citrus pulp ceased. Five calves whose dams had been fed citrus pulp were subsequently born with superior prognathism. In contrast to the eight cows that developed the syndrome only one out of 68 heifers which were fed larger quantities of citrus pulp for 10 days developed mild signs of the syndrome and then recovered, suggesting that older animals may be more susceptible. The clinical signs, gross pathology and histopathology are described and compared with those of previous outbreaks. Mycotoxins, particularly citrinin, were strongly implicated as the cause of this outbreak.


Aromatic C-F Hydroxylation by Nonheme Iron(IV)-Oxo Complexes: Structural, Spectroscopic, and Mechanistic Investigations.
The synthesis and reactivity of a series of mononuclear nonheme iron complexes that carry out intramolecular aromatic C-F hydroxylation reactions is reported. The key intermediate prior to C-F hydroxylation, [FeIV(O)(N4Py2Ar1)](BF4)2 (1-O, Ar1 = -2,6-difluorophenyl), was characterized by single-crystal X-ray diffraction. The crystal structure revealed a nonbonding C-H···O═Fe interaction with a CH3CN molecule. Variable-field Mössbauer spectroscopy of 1-O indicates an intermediate-spin (S = 1) ground state. The Mössbauer parameters for 1-O include an unusually small quadrupole splitting for a triplet FeIV(O) and are reproduced well by density functional theory calculations. With the aim of investigating the initial step for C-F hydroxylation, two new ligands were synthesized, N4Py2Ar2 (L2, Ar2 = -2,6-difluoro-4-methoxyphenyl) and N4Py2Ar3 (L3, Ar3 = -2,6-difluoro-3-methoxyphenyl), with -OMe substituents in the meta or ortho/para positions with respect to the C-F bonds. FeII complexes [Fe(N4Py2Ar2)(CH3CN)](ClO4)2 (2) and [Fe(N4Py2Ar3)(CH3CN)](ClO4)2 (3) reacted with isopropyl 2-iodoxybenzoate to give the C-F hydroxylated FeIII-OAr products. The FeIV(O) intermediates 2-O and 3-O were trapped at low temperature and characterized. Complex 2-O displayed a C-F hydroxylation rate similar to that of 1-O. In contrast, the kinetics (via stopped-flow UV-vis) for complex 3-O displayed a significant rate enhancement for C-F hydroxylation. Eyring analysis revealed the activation barriers for the C-F hydroxylation reaction for the three complexes, consistent with the observed difference in reactivity. A terminal FeII(OH) complex (4) was prepared independently to investigate the possibility of a nucleophilic aromatic substitution pathway, but the stability of 4 rules out this mechanism. Taken together the data fully support an electrophilic C-F hydroxylation mechanism.


Evolution Pathway from Iron Compounds to Fe1(II)-N4 Sites through Gas-Phase Iron during Pyrolysis.
Pyrolysis is indispensable for synthesizing highly active Fe-N-C catalysts for the oxygen reduction reaction (ORR) in acid, but how Fe, N, and C precursors transform to ORR-active sites during pyrolysis remains unclear. This knowledge gap obscures the connections between the input precursors and the output products, clouding the pathway toward Fe-N-C catalyst improvement. Herein, we unravel the evolution pathway of precursors to ORR-active catalyst comprised exclusively of single-atom Fe1(II)-N4 sites via in-temperature X-ray absorption spectroscopy. The Fe precursor transforms to Fe oxides below 300 °C and then to tetrahedral Fe1(II)-O4 via a crystal-to-melt-like transformation below 600 °C. The Fe1(II)-O4 releases a single Fe atom that diffuses into the N-doped carbon defect forming Fe1(II)-N4 above 600 °C. This vapor-phase single Fe atom transport mechanism is verified by synthesizing Fe1(II)-N4 sites via "noncontact pyrolysis" wherein the Fe precursor is not in physical contact with the N and C precursors during pyrolysis.


Distinct regulation of c-myb gene expression by HoxA9, Meis1 and Pbx proteins in normal hematopoietic progenitors and transformed myeloid cells.
The proto-oncogenic protein c-Myb is an essential regulator of hematopoiesis and is frequently deregulated in hematological diseases such as lymphoma and leukemia. To gain insight into the mechanisms underlying the aberrant expression of c-Myb in myeloid leukemia, we analyzed and compared c-myb gene transcriptional regulation using two cell lines modeling normal hematopoietic progenitor cells (HPCs) and transformed myelomonocytic blasts. We report that the transcription factors HoxA9, Meis1, Pbx1 and Pbx2 bind in vivo to the c-myb locus and maintain its expression through different mechanisms in HPCs and leukemic cells. Our analysis also points to a critical role for Pbx2 in deregulating c-myb expression in murine myeloid cells cotransformed by the cooperative activity of HoxA9 and Meis1. This effect is associated with an intronic positioning of epigenetic marks and RNA polymerase II binding in the orthologous region of a previously described alternative promoter for c-myb. Taken together, our results could provide a first hint to explain the abnormal expression of c-myb in leukemic cells.


Vanadyl as a Stable Structural Mimic of Reactive Ferryl Intermediates in Mononuclear Nonheme-Iron Enzymes.
The iron(II)- and 2-(oxo)glutarate-dependent (Fe/2OG) oxygenases catalyze an array of challenging transformations via a common iron(IV)-oxo (ferryl) intermediate, which in most cases abstracts hydrogen (H•) from an aliphatic carbon of the substrate. Although it has been shown that the relative disposition of the Fe-O and C-H bonds can control the rate of H• abstraction and fate of the resultant substrate radical, there remains a paucity of structural information on the actual ferryl states, owing to their high reactivity. We demonstrate here that the stable vanadyl ion [(VIV-oxo)2+] binds along with 2OG or its decarboxylation product, succinate, in the active site of two different Fe/2OG enzymes to faithfully mimic their transient ferryl states. Both ferryl and vanadyl complexes of the Fe/2OG halogenase, SyrB2, remain stably bound to its carrier protein substrate (l-aminoacyl-S-SyrB1), whereas the corresponding complexes harboring transition metals (Fe, Mn) in lower oxidation states dissociate. In the well-studied taurine:2OG dioxygenase (TauD), the disposition of the substrate C-H bond relative to the vanadyl ion defined by pulse electron paramagnetic resonance methods is consistent with the crystal structure of the reactant complex and computational models of the ferryl state. Vanadyl substitution may thus afford access to structural details of the key ferryl intermediates in this important enzyme class.


Plant tropane alkaloid biosynthesis evolved independently in the Solanaceae and Erythroxylaceae.
The pharmacologically important tropane alkaloids have a scattered distribution among angiosperm families, like many other groups of secondary metabolites. To determine whether tropane alkaloids have evolved repeatedly in different lineages or arise from an ancestral pathway that has been lost in most lines, we investigated the tropinone-reduction step of their biosynthesis. In species of the Solanaceae, which produce compounds such as atropine and scopolamine, this reaction is known to be catalyzed by enzymes of the short-chain dehydrogenase/reductase family. However, in Erythroxylum coca (Erythroxylaceae), which accumulates cocaine and other tropane alkaloids, no proteins of the short-chain dehydrogenase/reductase family were found that could catalyze this reaction. Instead, purification of E. coca tropinone-reduction activity and cloning of the corresponding gene revealed that a protein of the aldo-keto reductase family carries out this reaction in E. coca. This protein, designated methylecgonone reductase, converts methylecgonone to methylecgonine, the penultimate step in cocaine biosynthesis. The protein has highest sequence similarity to other aldo-keto reductases, such as chalcone reductase, an enzyme of flavonoid biosynthesis, and codeinone reductase, an enzyme of morphine alkaloid biosynthesis. Methylecgonone reductase reduces methylecgonone (2-carbomethoxy-3-tropinone) stereospecifically to 2-carbomethoxy-3β-tropine (methylecgonine), and has its highest activity, protein level, and gene transcript level in young, expanding leaves of E. coca. This enzyme is not found at all in root tissues, which are the site of tropane alkaloid biosynthesis in the Solanaceae. This evidence supports the theory that the ability to produce tropane alkaloids has arisen more than once during the evolution of the angiosperms.