New Insights into the Cystine-Sulfite Reaction.
The mechanism by which cysteine-S-sulfate is formed from the reaction of sulfite with cystine in the absence of a dedicated oxidizing agent is investigated using high-resolution NMR. Changes to reactant ratio, pH, UV light exposure and temperature were evaluated to determine the most effective conditions to achieve the maximum yield of cysteine-S-sulfate without recourse to conventional oxidizing reagents. Herein evidence is provided for both nucleophilic and radical mechanisms, by which cysteine-S-sulfate can be generated with yields of up to 96%.


Hematopoietic stem cell transplantation in the treatment of lymphoma.
In 1990, high-dose chemotherapy, with or without radiotherapy, requiring the support of hematopoietic stem cell transplantation has been confirmed as an effective treatment modality for a fraction of recurring or refractory lymphomas. Most relevant clinical studies published in 1990 are summarized and commented on in this review. Upcoming developments expected to improve the therapeutic index of high-dose therapies comprise the identification of patients at high risk of relapse so they can be treated earlier in the course of the disease, and the utilization of hematopoietic growth factors for ameliorating myelosuppression, as well as for harvesting peripheral blood hematopoietic stem cells utilizable for transplantation.


Electrophysiological dissociation of rapid memory mechanisms in humans.
Human memory involves an interaction between transient working memory and a long-term store. We found electrophysiological evidence supporting the existence of two distinct mechanisms subserving these processes. Stimuli held in memory for less than 4 s generate large, early latency P300 potentials which may index activation of a frontally-mediated rapid working memory system. Stimuli held in memory for over 4 s selectively generate N400 potentials which may reflect activation of mesial temporal cortices involved in access to the long-term store. These results suggest that memory processing in the initial 10 s after stimulus detection involves at least two distinct distributed cortical-limbic systems.


Acute penetrating injury of the spinal cord by a wooden spike with delayed surgery: a case report.
Rarely, penetrating injuries to the spinal cord result from wooden objects, creating unique challenges to mitigate neurological injury and high rates of infection and foreign body reactions. We report a man who sustained a penetrating cervical spinal cord injury from a sharpened stick. While initially tetraparetic, he rapidly recovered function. The risks of neurological deterioration during surgical removal made the patient reluctant to consent to surgery despite the impalement of the spinal cord. A repeat MRI on day 3 showed an extension of edema indicating progressive inflammation. On the 7th day after injury, fever and paresthesias occurred with a large increase in serum inflammatory indicators, and the patient agreed to undergo surgical removal of the wooden object. We discuss the management nuances related to wood, the longitudinal evolution of MRI findings, infection risk, surgical risk mitigation and technique, an inflammatory marker profile, long-term recovery, and the surprisingly minimal neurological deficits associated with low-velocity midline spinal cord injuries. The patient had an excellent clinical outcome. The main lessons are that a wooden penetrating central nervous system injury has a high risk for infection, and that surgical removal from the spinal cord should be performed soon after injury and under direct visualization.


Noninvasive assessment for acute allograft rejection in a rat lung transplantation model.
After lung transplantation, early detection of acute allograft rejection is important not only for timely and optimal treatment, but also for the prediction of chronic rejection which is a major cause of late death. Many biological and immunological approaches have been developed to detect acute rejection; however, it is not well known whether lung mechanics correlate with disease severity, especially with pathological rejection grade. In this study, we examined the relationship between lung mechanics and rejection grade development in a rat acute rejection model using the forced oscillation technique, which provides noninvasive assessment of lung function. To this end, we assessed lung resistance and elastance (RL and EL) from implanted left lung of these animals. The perivascular/interstitial component of rejection severity grade (A-grade) was also quantified from histological images using tissue fraction (TF; tissue + cell infiltration area/total area). We found that TF, RL, and EL increased according to A-grade. There was a strong positive correlation between EL at the lowest frequency (Elow; EL at 0.5 Hz) and TF (r(2) = 0.930). Furthermore, the absolute difference between maximum value of EL (Emax) and Elow (Ehet; Emax - Elow) showed the strong relationship with standard deviation of TF (r(2) = 0.709), and A-grade (Spearman's correlation coefficients; rs = 0.964, P < 0.0001). Our results suggest that the dynamic elastance as well as its frequency dependence have the ability to predict A-grade. These indexes should prove useful for noninvasive detection and monitoring the progression of disease in acute rejection.


Verapamil Use Is Associated With Reduction of Newly Diagnosed Diabetes Mellitus.
The mechanism of the beneficial effect of calcium-channel blockers (CCBs), especially verapamil, on the development of type 2 diabetes mellitus (T2DM) has been described. This study compared the incidence of T2DM in adults prescribed oral verapamil and propensity score-matched adults prescribed other oral CCBs. This retrospective population-based cohort study used Taiwan's National Health Insurance Research Database from 2000 to 2011. T2DM was defined according to the International Classification of Diseases, Ninth Revision, Clinical Modification. During follow-up periods of 41,958 and 42,118 person-years, 269 of 4930 patients in the verapamil cohort and 340 of 4930 patients in the matched cohort, respectively, developed T2DM. The incidence rates were 6.41 and 8.07 per 1000 population per year among verapamil and other CCB users, respectively. The adjusted hazard ratio (HR) for T2DM associated with use of verapamil (vs. other CCBs) was 0.80 [95% confidence interval (CI), 0.68 to 0.94; P = 0.006]. After exclusion of patients followed for <180 days or <365 days (to avoid bias derived from delayed diagnosis), adjusted HRs remained significant [0.79 (95% CI, 0.67 to 0.93; P = 0.005) and 0.77 (95% CI, 0.65 to 0.91; P = 0.002), respectively]. Only the interaction term for age was significant (P = 0.009). Verapamil had a more prominent effect on patients aged older than 65 years (P < 0.001). In patients with no known history of diabetes mellitus, oral verapamil use was associated with a decreased incidence of T2DM compared with other CCBs.


Expression of bcl-2 in classical Hodgkin's lymphoma: an independent predictor of poor outcome.
Although most classical Hodgkin's lymphoma (CHL) patients are cured, a significant minority fails primary therapy and may die as a result of their disease. Age, stage, and other basic clinical and laboratory parameters, which comprise the International Prognostic Score (IPS), are used at diagnosis to predict outcome. To date, there is no consensus on biologic markers that add value to these parameters. We evaluated 107 CHL patients for bcl-2, p53, and p21 expression by immunohistochemistry using tissue microarrays and correlated the results with outcome. The median follow-up of the 79 surviving patients was 6.8 years. Univariate analysis showed that age > or = 45 years, stage III or IV, and IPS > or = 3 were associated with a poor failure-free survival (FFS) and overall survival (OS). bcl-2 was expressed in 26% of patients and was associated with poor FFS and a trend for OS. p53 expression in combination with lack of p21 expression was not associated with outcome. Multivariate analysis showed that three factors were independently associated with both FFS and OS: age > or = 45 years, stage III or IV, and bcl-2 expression. Using these three parameters, a scoring system was devised that stratified patients into three risk groups (with zero, one, or two to three of these risk factors) and a progressively worse FFS and OS (P < .001). Expression of bcl-2 in CHL is a useful, independent prognostic marker and can be used in association with clinical parameters to identify newly diagnosed patients with a good, intermediate, or poor prognosis.


Metabolic parameters: how important are pharmacologically-induced changes?
It has been suggested that beta-blockers do not significantly reduce the frequency of coronary disease in hypertensives because they adversely affect the plasma lipoprotein profile. Non-selective beta-blockers cause a significant elevation of triglycerides and a reduction in the level of high-density lipoprotein. Cardioselective beta-blockers have similar, but less marked, effects. The non-selective beta-blockers with intrinsic sympathomimetic activity have little effect on the lipid profile, yet these are the least effective in reducing coronary mortality after myocardial infarction. Elevated blood flow velocity and high catecholamine levels can damage the vascular endothelium. This leads to increased permeability and the deposition of lipids. Beta-blockers reduce peak blood velocity and decrease the activity of catecholamines in plasma, which should reduce endothelial damage and the subsequent formation of plaques. Studies in animals have indeed shown that beta-blockers reduce the level of atherosclerosis, despite an elevation of serum lipids.


Binding of a designed substrate analogue to diisopropyl fluorophosphatase: implications for the phosphotriesterase mechanism.
A wide range of organophosphorus nerve agents, including Soman, Sarin, and Tabun is efficiently hydrolyzed by the phosphotriesterase enzyme diisopropyl fluorophosphatase (DFPase) from Loligo vulgaris. To date, the lack of available inhibitors of DFPase has limited studies on its mechanism. The de novo design, synthesis, and characterization of substrate analogues acting as competitive inhibitors of DFPase are reported. The 1.73 A crystal structure of O,O-dicyclopentylphosphoroamidate (DcPPA) bound to DFPase shows a direct coordination of the phosphoryl oxygen by the catalytic calcium ion. The binding mode of this substrate analogue suggests a crucial role for electrostatics in the orientation of the ligand in the active site. This interpretation is further supported by the crystal structures of double mutants D229N/N120D and D229N/N175D, designed to reorient the electrostatic environment around the catalytic calcium. The structures show no differences in their calcium coordinating environment, although they are enzymatically inactive. Additional double mutants E21Q/N120D and E21Q/N175D are also inactive. On the basis of these crystal structures and kinetic and mutagenesis data as well as isotope labeling we propose a new mechanism for DFPase activity. Calcium coordinating residue D229, in concert with direct substrate activation by the metal ion, renders the phosphorus atom of the substrate susceptible for attack of water, through generation of a phosphoenzyme intermediate. Our proposed mechanism may be applicable to the structurally related enzyme paraoxonase (PON), a component of high-density lipoprotein (HDL).


Where treatment and prevention merge: the need for a broader approach.
Four papers were presented in a Parallel Session chaired by Dr Forge Gleser (Israel) and these are summarized here. The papers were: (1) "Supporting primary health care physicians" by Dr Galina A. Korchagina of the Medical Academy of Postgraduate Studies, St Petersburg, the Russian Federation; (2) "A new role for nurses" by Ms Iveta Kelle, Nursing Consultant, Riga, Latvia; (3) "Broadening the base of treatment" by Dr Nina Kerimi of the Research Institute of Preventive and Clinical Medicine, Ashgabat, Turkmenistan; and (4) "Primary health care and alcohol reduction strategies" by Dr Leo Pas of the Academic Centre of General Practice, Free University Brussels, Belgium. Following a discussion, participants agreed on the wording of a draft Policy for Primary Health Care and Community Action and this is also included here.


[Abdominal intercostal neuralgia: a forgotten cause of abdominal pain].
Four patients, 3 women aged 39, 36 and 58 and a man aged 51, had been experiencing recurring bouts of abdominal pain for periods varying from a few months to years. The pain could be provoked by palpation of a small circumscript area in the lower abdomen. A positive Carnett's sign (pain intensification during palpation while contracting the abdominal muscles by raising the head, whilst lying flat) may aid the diagnosis. An injection ofa local anaesthetic agent confirmed the diagnosis of nerve entrapment and appeared therapeutic in one patient. The other three patients experienced long-term relief following surgical nerve excision. Establishing a diagnosis in patients with abdominal cutaneous nerve entrapment syndrome is often delayed due to physicians being unaware of this condition. Most patients have to undergo numerous investigations including laparoscopies and explorative laparotomies, often to no avail. This syndrome should be considered in patients with chronic abdominal pain syndromes.


Reconceptualising Atrial Fibrillation Using Renewal Theory: A Novel Approach to the Assessment of Atrial Fibrillation Dynamics.
Despite a century of research, the mechanisms of AF remain unresolved. A universal motif within AF research has been unstable re-entry, but this remains poorly characterised, with competing key conceptual paradigms of multiple wavelets and more driving rotors. Understanding the mechanisms of AF is clinically relevant, especially with regard to treatment and ablation of the more persistent forms of AF. Here, the authors outline the surprising but reproducible finding that unstable re-entrant circuits are born and destroyed at quasi-stationary rates, a finding based on a branch of mathematics known as renewal theory. Renewal theory may be a way to potentially unify the multiple wavelet and rotor theories. The renewal rate constants are potentially attractive because they are temporally stable parameters of a defined probability distribution (the exponential distribution) and can be estimated with precision and accuracy due to the principles of renewal theory. In this perspective review, this new representational architecture for AF is explained and placed into context, and the clinical and mechanistic implications are discussed.


Chlorpheniramine and escitalopram: Similar antidepressant and nitric oxide lowering roles in a mouse model of anxiety.
There is a crosstalk between mood disorders and oxidative stress. Chlorpheniramine (CPA), a first generation antihistamine, is hypothesized to have an anxiolytic role at high doses; however, its antidepressant and antioxidant roles have not previously been investigated. The aim of the current study was to evaluate the antidepressant and anxiolytic effects of CPA treatment in association with nitric oxide (NO) and super oxide dismutase (SOD) activity in a mouse model of anxiety. BALB/c mice were divided into unstressed (naïve), control, and CPA- (0.5 mg/kg) and escitalopram- (ESC; 10 mg/kg) treated groups for 3 weeks. Subsequently, they were immobilized for 6 h and subjected to behavioural paradigms as follows: The open field test, the elevated plus maze (EPM) and the forced swim test to investigate motor function, anxiety and depression, respectively. The mice were sacrificed and serum was obtained to detect NO and SOD activity. Compared with the control group, the CPA-treated group demonstrated an antidepressant effect similar to that of the ESC-treated group. In addition, CPA prevented stress-induced NO without affecting SOD activity. CPA did not improve anxiety-like behaviour in the EPM, nor did it improve stress-induced locomotion and rearing, as demonstrated by the OFT. Thus, to the best of our knowledge, this is the first study to evaluate the antidepressant role of CPA in association with NO metabolism. However, further studies are required to elucidate the underlying mechanism.


R56865, a potent new antiarrhythmic agent, effective during ischemia and reperfusion in the rat heart.
The antiarrhythmic effects of R56865 were characterized both in vivo and in vitro. Four groups (n = 12 per group) of anesthetized rats, subjected to 5- or 30-min coronary artery ligation and reperfusion, were studied: saline, dimethyl sulfoxide (DMSO) carrier, and R56865 (0.5 or 2 mg/kg) were administered as an intravenous (i.v.) bolus before ligation. After 5 min of ischemia, the incidences of reperfusion-induced ventricular tachycardia (VT) and fibrillation (VF), which were high in the saline (100 and 75%, respectively) and DMSO (100 and 82%, respectively) control groups, were abolished with both doses of R56865. With 30 min of ischemia, R56865 (2 mg/kg) significantly reduced the incidences of ischemia-induced VT and VF (from 100 and greater than 50% to 25 and 8%, respectively). For in vitro studies, five groups (n = 12 per group) of isolated rat hearts subjected to 10- or 30-min coronary ligation and reperfusion were studied: unmodified buffer and buffer containing DMSO or R56865 (10(-7), 10(-8), 10(-9) M). After 10 min of ischemia, R56865 (10(-7) M) decreased reperfusion-induced VT and VF (from 100 and 75% in buffer controls to 42 and 8%, respectively) when administered throughout the experiment. With 30 min of ischemia, R5685 (10(-7) M) reduced the incidences of ischemia-induced VT and VF (from 75 and 67% in the buffer controls to 25 and 25%, respectively). Although reperfusion after 30 min of ischemia did not induce VF in any of the groups studied, VT and other arrhythmias did occur and their incidences were reduced significantly by R56865. To investigate whether calcium overload might mediate the effects of R56865, hearts were perfused aerobically with a high-calcium/low-sodium medium. VT and VF occurred in 80% of control hearts; R56865 (10(-7) M) did not prevent these arrhythmias. In conclusion, R56865 exerts a potent effect against ischemia- and reperfusion-induced arrhythmias through a mechanism which appears to operate during ischemia.


Early events in insulin fibrillization studied by time-lapse atomic force microscopy.
The importance of understanding the mechanism of protein aggregation into insoluble amyloid fibrils lies not only in its medical consequences, but also in its more basic properties of self-organization. The discovery that a large number of uncorrelated proteins can form, under proper conditions, structurally similar fibrils has suggested that the underlying mechanism is a general feature of polypeptide chains. In this work, we address the early events preceding amyloid fibril formation in solutions of zinc-free human insulin incubated at low pH and high temperature. Here, we show by time-lapse atomic force microscopy that a steady-state distribution of protein oligomers with a quasiexponential tail is reached within a few minutes after heating. This metastable phase lasts for a few hours, until fibrillar aggregates are observable. Although for such complex systems different aggregation mechanisms can occur simultaneously, our results indicate that the prefibrillar phase is mainly controlled by a simple coagulation-evaporation kinetic mechanism, in which concentration acts as a critical parameter. These experimental facts, along with the kinetic model used, suggest a critical role for thermal concentration fluctuations in the process of fibril nucleation.


Association of Time to Definitive Hemostasis With Mortality in Patients With Solid Organ Injuries.
Introduction The Golden Hour is a term used in the trauma setting to refer to the first 60 minutes after injury. Traditionally, definitive care within this period was believed to dramatically increase a patient's survival. Though the period of 60 minutes is unlikely to represent a point of distinct inflection in survival, the effect of time to definitive care on survival remains incompletely understood. This study aims to measure the association of time to definitive hemostasis with mortality in patients with solid organ injuries as well as the effect of survival bias and a form of selection bias known as indication by severity on the relationship between time to treatment and survival. Methodology This is a retrospective cohort study using data obtained from the American College of Surgeons National Trauma Data Bank (NTDB) from the years 2017 through 2019 selecting patients treated for blunt liver, spleen, or kidney injury who required angioembolization or surgical hemostasis within six hours. A Cox proportional hazards regression was used to analyze time to death. The association of probability of death with time was examined with a multivariate logistic regression initially treating the relationship as linear and subsequently transforming time to hemostasis with restricted cubic splines to model a non-linear association with the outcome. To model survival and indication by severity bias, we created a computer-generated data set and used LOESS regressions to display curves of the simulated data. Results The multivariate Cox proportional hazards analysis shows a coefficient of negative 0.004 for minutes to hemostasis with an adjusted hazard ratio of 0.9959 showing the adjusted hazard of death slightly diminishes with each increasing minute to hemostasis. The likelihood ratio chi-square difference between the model with time to hemostasis included as a linear term versus the model with the restricted cubic spline transformation is 97.46 (p<0.0001) showing the model with restricted cubic splines is a better fit for the data. The computer-generated data simulating treatment of solid organ injury with no programmed bias displays an almost linear association of mortality with increased treatment delay. When indications by severity bias and survival bias are introduced, the risk of death decreases with time to hemostasis as in the real-world data. Conclusion Decreasing mortality with increasing delay to hemostasis in trauma patients with solid organ injury is likely due to confounding due to indication by severity and survival bias. After taking these biases into account, the association of delayed hemostasis with better survival is not likely due to the benefit of delay but rather the delay sorts patients by severity of injury with those more likely to die being treated first. These biases are extremely difficult to eliminate which limits the ability to measure the true effect of delay with retrospective data. The findings may however be of value as a predictive model to anticipate the acuity of a patient after an interval of unavoidable delay such as with a long transfer time.


Artificial cell membrane camouflaged immunomagnetic nanoparticles for enhanced circulating tumor cell isolation.
Precise and specific circulating tumor cell (CTC) isolation is heavily interfered with by blood cells and proteins. Although satisfactory results have been achieved by some cell membrane-derived platforms, the following limitations have seriously limited the commercialization potential: complex membrane composition, difficult batch difference control, inconvenient source cell expansion, etc. To overcome these limitations, artificial cell membrane camouflage made from commercialized lipids and proteins was proposed in this work. Specifically, a biotinylated phospholipid which can serve as a lipid component and provide active sites (biotin) for antibody modification, and human serum albumin (HSA), which can effectively reduce certain blood protein adsorption, were applied simultaneously to endow our immunomagnetic platform with a new biological identity. Besides, making full use of the robust lipid and protein absorption ability of graphene nanosheets (GNs), the biomimetic cell membrane can be easily integrated into the magnetic core through simple lipid and protein solution incubation. Surprisingly, the resulting artificial cell membrane camouflaged immunomagnetic nanoparticles (AIMNPs) achieved high specificity (average capture efficiency 87.0%), good sensitivity (7 model CTCs per 0.5 mL) and an enhanced anti-nonspecific absorption ability (15-105 white blood cells per 1.5 mL) in both mimic and clinical blood samples.


Restless legs syndrome in hemodialysis patients: health-related quality of life and laboratory data analysis.
To compare clinical data, sleep quality and health-related quality of life (HRQOL) with and without RLS in HD patients. The international RLS study group diagnosis questionnaire was completed by 228 HD patients. The Pittsburg Sleep Quality Index (PSQI) for the evaluation of sleep quality and the Kidney Disease Quality of Life (KDQOL-SF) for the analysis of HRQOL were also used. 53 (23%) patients were diagnosed as RLS. Age and age at the initiation of HD were significantly younger in the RLS group. Serum calcium concentration (Ca) was significantly higher in the RLS group. Sleep quality evaluated by PSQI was significantly lower in the RLS group. In SF-36 domains of KDQOL-SF, bodily pain, general health perceptions, vitality, role functioning emotional, mental health and mental component score were significantly lower in the RLS group. In kidney targeted scales of KDQOL-SF, symptoms/problems, burden of kidney disease, cognitive function, quality of social interaction, sleep and patient satisfaction were significantly lower in the RLS group. High Ca was possibly connected to the pathophysiology of RLS which impaired sleep quality as well as HRQOL including mental health and many kidney disease related scales.


4,4'-diaponeurosporene, a C30 carotenoid, effectively activates dendritic cells via CD36 and NF-κB signaling in a ROS independent manner.
Carotenoids could be divided into C30 carotenoids and C40 carotenoids. The immune functions of C40 carotenoids had been extensively researched, while those of C30 carotenoids still remain unclear. In this study, the immune functions of a biosynthetic C30 carotenoid, 4,4'-diaponeurosporene (Dia), were identified on dendritic cells (DCs). DCs treated with 1 μM Dia for 24 h showed morphologic and phenotypic characteristics of mature state and had an increased production of IL-6, IL-10, IL-12p70 and TNFα, while β-carotene had a suppressive effect on DCs maturation. Moreover, Dia promoted antigen uptake of DCs in vitro and increased the quantity of antigen loaded DCs in mesenteric lymph nodes (MLN). Dia-treated DCs also had an enhanced ability to stimulate T cell proliferation and Th1 polarization. Further researches showed that Dia activated DCs via CD36 as well as ERK, JNK, and NF-κB signals in a reactive oxygen species (ROS) independent manner.


Bending Analysis of Asymmetric Functionally Graded Material Sandwich Plates in Thermal Environments.
This paper investigates the bending of asymmetric functionally graded material (FGM) sandwich plates subjected to thermo-mechanical loads in thermal environments. In this paper, a thermo-mechanical analysis model for asymmetric FGM sandwich plates is proposed, which contains only four control equations and four unknown variables. The governing equation is obtained through refined shear theory and the principle of virtual work, and the Navier method is used to solve it. Numerical examples of simply supported FGM sandwich plates under thermo-mechanical loads are given to verify the accuracy of the model. Finally, detailed studies are conducted on the bending of asymmetric FGM sandwich plates under thermo-mechanical loads, exploring the effects of various parameter changes on their bending behavior, and providing strong guidance for the application of asymmetric FGM sandwich plates in industrial production practice.


Applying lexical and semantic analysis to the exploration of free-text data.
A substantial study of primary nursing involving the authors ( 1 , 2 ) generated, in addition to the pre-coded data, a large number of free-text responses. We decided to investigate whether these less structured data could help to categorise and understand the leadership styles of ward sisters in the sample.


Porous, Ventricular Extracellular Matrix-Derived Foams as a Platform for Cardiac Cell Culture.
To more closely mimic the native cellular microenvironment, 3D scaffolds derived from the extracellular matrix (ECM) are being developed as alternatives to conventional 2D culture systems. In the present study, we established methods to fabricate nonchemically cross-linked 3D porous foams derived entirely from decellularized porcine left ventricle (DLV) for use as an in vitro cardiac cell culture platform. Furthermore, we explored the effects of physically preprocessing the DLV through mechanical mincing versus cryomilling, as well as varying the ECM concentration on the structure, composition, and physical properties of the foams. Our results indicate that the less highly processed minced foams had a more cohesive and complex network of ECM components, enhanced mechanical properties, and improved stability under simulated culturing conditions. To validate the DLV foams, a proof-of-concept study was conducted to explore the early cardiomyogenic differentiation of pericardial fat adipose-derived stem/stromal cells (pfASCs) on the minced DLV foams relative to purified collagen I gel controls. Differentiation was induced using a modified cardiomyogenic medium (MCM) or through stimulation with 5-azacytidine (5-aza), and cardiomyocyte marker expression was characterized by immunohistochemistry and real-time reverse transcriptase-polymerase chain reaction. Our results indicate that early markers of cardiomyogenic differentiation were significantly enhanced on the DLV foams cultured in MCM, suggesting a synergistic effect of the cardiac ECM-derived scaffolds and the culture medium on the induction of pfASC differentiation. Furthermore, in analyzing the response in the noninduced control groups, the foams were observed to provide a mildly inductive microenvironment for pfASC cardiomyogenesis, supporting the rationale for using tissue-specific ECM as a substrate for cardiac cell culture applications.


Effects of mutations in the beet western yellows virus readthrough protein on its expression and packaging and on virus accumulation, symptoms, and aphid transmission.
Virions of beet western yellows luteovirus contain a major capsid protein (P22.5) and a minor readthrough protein (P74), produced by translational readthrough of the major capsid protein sequence into the neighboring open reading frame, which encodes the readthrough domain (RTD). The RTD contains determinants required for efficient virus accumulation in agroinfected plants and for aphid transmission. The C-terminal halves of the RTD are not well conserved among luteoviruses but the N-terminal halves contain many conserved sequence motifs, including a proline-rich sequence separating the rest of the RTD from the sequence corresponding to the major coat protein. To map different biological functions to these regions, short in-frame deletions were introduced at different sites in the RTD and the mutant genomes were transmitted to protoplasts as transcripts and to Nicotiana clevelandii by agroinfection. Deletions in the nonconserved portion of the RTD did not block aphid transmission but had a moderate inhibitory effect on virus accumulation in plants and abolished symptoms. Deletion of the proline tract and the junction between the conserved and nonconserved regions inhibited readthrough protein accumulation in protoplasts by at least 10-fold. The mutants accumulated small amounts of virus in plants, did not induce symptoms, and were nontransmissible by aphids using agroinfected plants, extracts of infected protoplasts, or purified virus as a source of inoculum. Other deletions in the conserved portion of the RTD did not markedly diminish readthrough protein accumulation but abolished its incorporation into virions. These mutants accumulated to low levels in agroinfected plants and elicited symptoms, but could not be aphid-transmitted. A preliminary map has been produced mapping these functions to different parts of the RTD.


Transient equilibrium determination of dopamine D2/D3 receptor densities and affinities in brain.
Long-term alteration of dopaminergic neurotransmission is known to modulate the D2/D3 receptor expression in the brain. The modulation can occur as a response to pathological processes or pharmacological intervention. The receptor density can be monitored by in vivo positron emission tomography (PET) of [11C] raclopride. To obtain accurate measurements of receptor-ligand interaction, it is essential to estimate binding parameters at true (if transient) equilibrium of bound and unbound ligand quantities. We designed this study as a comparison of two quantitative approaches to transient equilibrium, the TRansient EquilibriuM BoLus Estimation (TREMBLE) method and the Transient Equilibrium Model (TEM) method, to determine binding parameters at transient equilibrium with bolus injection of the radioligand. The data demonstrates that TREMBLE unlike TEM identified the time at which equilibrium existed. TREMBLE revealed that equilibrium prevailed at one or more times after bolus injection and identified differences of receptor density among regions such as putamen and caudate nucleus. We demonstrated that TREMBLE is a quantitative approach suitable for the study of pathophysiological conditions of certain types of neurotransmission the brain.


Structure-activity relationships of antitubercular nitroimidazoles. 3. Exploration of the linker and lipophilic tail of ((s)-2-nitro-6,7-dihydro-5H-imidazo[2,1-b][1,3]oxazin-6-yl)-(4-trifluoromethoxybenzyl)amine (6-amino PA-824).
The (S)-2-nitro-6-(4-(trifluoromethoxy)benzyloxy)-6,7-dihydro-5H-imidazo[2,1-b][1,3]oxazine named PA-824 (1) has demonstrated antitubercular activity in vitro and in animal models and is currently in clinical trials. We synthesized derivatives at three positions of the 4-(trifluoromethoxy)benzylamino tail, and these were tested for whole-cell activity against both replicating and nonreplicating Mycobacterium tuberculosis (Mtb). In addition, we determined their kinetic parameters as substrates of the deazaflavin-dependent nitroreductase (Ddn) from Mtb that reductively activates these pro-drugs. These studies yielded multiple compounds with 40 nM aerobic whole cell activity and 1.6 μM anaerobic whole cell activity: 10-fold improvements over both characteristics from the parent molecule. Some of these compounds exhibited enhanced solubility with acceptable stability to microsomal and in vivo metabolism. Analysis of the conformational preferences of these analogues using quantum chemistry suggests a preference for a pseudoequatorial orientation of the linker and lipophilic tail.


Complementation of biotransformations with chemical C-H oxidation: copper-catalyzed oxidation of tertiary amines in complex pharmaceuticals.
The isolation, quantitation, and characterization of drug metabolites in biological fluids remain challenging. Rapid access to oxidized drugs could facilitate metabolite identification and enable early pharmacology and toxicity studies. Herein, we compared biotransformations to classical and new chemical C-H oxidation methods using oxcarbazepine, naproxen, and an early compound hit (phthalazine 1). These studies illustrated the low preparative efficacy of biotransformations and the inability of chemical methods to oxidize complex pharmaceuticals. We also disclose an aerobic catalytic protocole (CuI/air) to oxidize tertiary amines and benzylic CH's in drugs. The reaction tolerates a broad range of functionalities and displays a high level of chemoselectivity, which is not generally explained by the strength of the C-H bonds but by the individual structural chemotype. This study represents a first step toward establishing a chemical toolkit (chemotransformations) that can selectively oxidize C-H bonds in complex pharmaceuticals and rapidly deliver drug metabolites.


Challenges in Managing Isolated Subsegmental Pulmonary Embolism.
This commentary explores the clinical conundrums arising when caring for patients with acute pulmonary embolism isolated to the subsegmental pulmonary arteries. We discuss ways to confirm the radiologic diagnosis, how to distinguish patients for whom anticoagulation is indicated from those who are eligible for structured surveillance without anticoagulation, what surveillance entails, and why ensuring continuity of care matters. We report a case from our own experience that illustrates these decision-making crossroads and highlights the importance of cross-disciplinary collaboration. Because the evidence in the literature is currently weak and indirect, we draw on expert opinion in US and European guidelines, a recent statement from a multidisciplinary consensus panel, and several ongoing well-designed clinical trials. This discussion will help clinicians better manage the spectrum of patients who present with isolated subsegmental embolism.


Oecomys catherinae (Sigmodontinae, Cricetidae): Evidence for chromosomal speciation?
Among the Oryzomyini (Sigmodontinae), Oecomys is the most speciose, with 17 species. This genus presents high karyotypic diversity (2n = 54 to 2n = 86) and many taxonomic issues at the species level because of the presence of cryptic species and the overlap of morphological characters. For these reasons the real number of species of Oecomys may be underestimated. With the aim of verifying if the taxon Oecomys catherinae is composed of more than one species, we made comparative studies on two populations from two regions of Brazil, one from the Amazon and another from the Atlantic Forest using both classical cytogenetics (G- and C-banding) and comparative genomic mapping with whole chromosome probes of Hylaeamys megacephalus (HME), molecular data (cytochrome b mitochondrial DNA) and morphology. Our results confirm that Oecomys catherinae occurs in the southeast Amazon, and reveal a new karyotype for the species (2n = 62, FNa = 62). The comparative genomic analysis with HME probes identified chromosomal homeologies between both populations and rearrangements that are responsible for the different karyotypes. We compared our results in Sigmodontinae genera with other studies that also used HME probes. These chromosomal differences together with the absence of consistent differentiation between the two populations on morphological and molecular analyses suggest that these populations may represent cryptic species.


OBSERVATIONS ON THE USE OF THE HEN AND RABBIT ISOLATED AURICLES FOR THE DETERMINATION OF INOTROPIC POTENCY.
The electrically driven, isolated auricles from the rabbit and the hen have been used to compare the inotropic potency of a bufodiene aglycone with ouabain. The results from the hen auricle permit conventional statistical treatment and show a good degree of reproducibility. Results of inotropic potencies and toxicity determinations on the two drugs are discussed.


Strain-promoted oxidation-controlled cyclooctyne-1,2-quinone cycloaddition (SPOCQ) for fast and activatable protein conjugation.
A main challenge in the area of bioconjugation is to devise reactions that are both activatable and fast. Here, we introduce a temporally controlled reaction between cyclooctynes and 1,2-quinones, induced by facile oxidation of 1,2-catechols. This so-called strain-promoted oxidation-controlled cyclooctyne-1,2-quinone cycloaddition (SPOCQ) shows a remarkably high reaction rate when performed with bicyclononyne (BCN), outcompeting the well-known cycloaddition of azides and BCN by 3 orders of magnitude, thereby allowing a new level of orthogonality in protein conjugation.


Association of indoor and outdoor short-term PM2.5 exposure with blood pressure among school children.
The association between particulate matter and children's increased blood pressure is inconsistent, and few studies have evaluated indoor exposure, accounting for time-activity. The present study aimed to examine the association between personal short-term exposure to PM2.5 and blood pressure in children. We conducted a panel study with up to three physical examinations during different seasons of 2018 (spring, summer, and fall) among 52 children. The indoor PM2.5 concentration was continuously measured at home and classroom of each child using indoor air quality monitors. The outdoor PM2.5 concentration was measured from the nearest monitoring station. We constructed a mixed effect model to analyze the association of short-term indoor and outdoor PM2.5 exposure accounting for time-activity of each participant with blood pressure. The average PM2.5 concentration was 34.3 ± 9.2 μg/m3 and it was highest in the spring. The concentration measured at homes was generally higher than that measured at outdoor monitoring station. A 10-μg/m3 increment of the up to previous 3-day mean (lag0-3) PM2.5 concentration was associated with 2.7 mmHg (95%CI = 0.8, 4.0) and 2.1 mmHg (95%CI = 0.3, 4.0) increases in systolic and diastolic blood pressure, respectively. In a panel study comprehensively evaluating both indoor and outdoor exposures, which enabled more accurate exposure assessment, we observed a statistically significant association between blood pressure and PM2.5 exposure in children.


A Genetic Investigation of Island Jersey Cattle, the Foundation of the Jersey Breed: Comparing Population Structure and Selection to Guernsey, Holstein, and United States Jersey Cattle.
For two centuries, Jersey cattle were exported globally, adapting to varying climates and production systems, yet the founding population remained genetically isolated on the Island of Jersey. The Island of Jersey formally allowed the importation of pure Jersey cattle in 2008. This study characterized the genetic variation of 49 popular bulls from the Island of Jersey born from 1964 to 2004 and compared them to 47 non-Island Jersey bulls and cows, primarily from the United States In addition, 21 Guernsey cattle derived from the Island of Guernsey and 71 Holstein cattle served as reference populations for genetic comparison. Cattle were genotyped on the Illumina BovineHD Beadchip producing 777,962 SNPs spanning the genome. Principal component analysis revealed population stratification within breed reflective of individual animal's continental origin. When compared to Holstein and Guernsey, all Jersey clustered together by breed. The Jersey breed demonstrated increased inbreeding in comparison to Holstein or Guernsey with slightly higher estimates of inbreeding coefficients and identity-by-descent. The Island and United States Jersey have relatively similar, yet statistically different inbreeding estimates despite vastly different population sizes and gene flow. Signatures of selection within Island Jersey were identified using genome-wide homozygosity association and marker-based FST that provided population informative single-nucleotide polymorphism (SNPs). Biological significance of the homozygosity association results identified multiple genes on chromosomes 5, 24, and 27, involved in immune function and cellular processes. Overall, genomic variation was identified between the Island and non-Island Jersey cattle producing population informative SNPs and differing runs of homozygosity (ROH) over immune regulation and metabolic genes. Results on inbreeding measures and ROH may reflect varying effective population size or differential selection with grazing systems promoting natural selection for traits such as parasite resistance, whereas confinement systems demonstrate a more intensive artificial selection. More broadly, differences in breed formation, particularly between the two Channel Island breeds, likely contributed to the variation in ROH and inbreeding. This research provides a reference for the Jersey breed based on the genetic foundation of the Island cattle as compared to the intensively selected United States cattle, and identifies regions of the genome for future investigation of immune regulation and metabolic processes.


Computer-aided prediction of RNA secondary structures.
A brief survey of computer algorithms that have been developed to generate predictions of the secondary structures of RNA molecules is presented. Two particular methods are described in some detail. The first utilizes a thermodynamic energy minimization algorithm that takes into account the likelihood that short-range folding tends to be favored over long-range interactions. The second utilizes an interactive computer graphic modelling algorithm that enables the user to consider thermodynamic criteria as well as structural data obtained by nuclease susceptibility, chemical reactivity and phylogenetic studies. Examples of structures for prokaryotic 16S and 23S ribosomal RNAs, several eukaryotic 5S ribosomal RNAs and rabbit beta-globin messenger RNA are presented as case studies in order to describe the two techniques. Anm argument is made for integrating the two approaches presented in this paper, enabling the user to generate proposed structures using thermodynamic criteria, allowing interactive refinement of these structures through the application of experimentally derived data.


MiR-590-3p suppresses hepatocellular carcinoma growth by targeting TEAD1.
MicroRNA signature is altered in different disease states including cancer, and some microRNAs act as oncogenes or tumor suppressors. MiR-590-3p has been shown to be involved in human cancer progression. However, its role in hepatocellular carcinoma remains unknown. In this study, miR-590-3p level was measured, and clinicopathological features were determined in hepatocellular carcinoma tissues. The function of miR-590-3p was examined in vitro and in vivo. Real-time reverse transcription polymerase chain reaction analysis demonstrated downregulation of miR-590-3p in hepatocellular carcinoma tissues, and its downregulation was associated with a poor overall survival of hepatocellular carcinoma patients. Ectopic expression of miR-590-3p promoted growth of hepatocellular carcinoma cells, whereas its depletion inhibited cell growth. Transcriptional enhancer activator domain 1 was identified as a validated miR-590-3p target. Upregulation of transcriptional enhancer activator domain 1 was found in hepatocellular carcinoma tissues and inversely correlated with miR-590-3p. Our results indicate a tumor suppressor role of miR-590-3p in hepatocellular carcinoma through targeting transcriptional enhancer activator domain 1 and suggest its use in the diagnosis and prognosis of liver cancer.


The hemodynamics of cognitive control: the level of concentration of oxygenated hemoglobin in the superior prefrontal cortex varies as a function of performance in a modified Stroop task.
The purpose of this study was to establish the relationship between the hemodynamic response of prefrontal cortex (PFC) and individual differences in cognitive control, as measured by a color-word interference task. Twenty-five healthy volunteers were observed through functional near infrared spectroscopy (fNIRS) while performing a modified Stroop paradigm. Mean concentration levels of oxygenated hemoglobin (oxy-Hb) were correlated with behavioral performance in the conflict task. Those with shorter reaction times had higher levels of oxy-Hb concentration in superior dorsolateral PFC. Our results are the first to show a positive correlation between behavioral performance and oxy-Hb levels in superior dorsolateral PFC in a cognitive conflict task. These results suggest that the availability of oxygen in the superior PFC, possibly linked to an increase in metabolism, may be related to attention level and effectiveness of cognitive control.


Evaporation of Inclined Drops: Formation of Asymmetric Ring Patterns.
Evaporation of colloidal drops on horizontal surfaces deposits the contained particles at the drop-edge producing radially symmetric ring-like stains. The symmetry in the particle deposition is broken when the drop is placed on a tilted surface due to the influence of gravity on the suspended particles and the drop itself. Using extremely small drops generated by electrospray, we explore cases where different mechanisms of particle transport dominate. We show that the asymmetric residues are formed as the gravity-induced effects compete with the capillary flow. Our results give a broad insight into the pattern formation of evaporating inclined drops.


Influence of sex on blood pressure and left ventricular mass in adolescents: the Hypertension in Pregnancy Offspring Study.
The aim of the present longitudinal study was to describe determinants of blood pressure (BP) and echocardiographically calculated left ventricular mass in adolescents with a variable risk of future hypertension. BP was measured and M-mode echocardiography was performed in 52 children, 29 male and 23 female, at a mean age of 12.6 +/- 1.5 years. A re-examination was performed after 5.6 years. Thirty-six children were born to mothers who had hypertension during pregnancy whereas 16 children were born after a normotensive pregnancy. During follow-up a difference in body weight (P < 0.01) and height (P < 0.001) developed between male and female subjects. At the same time a difference in systolic blood pressure (SBP) appeared between sexes (P < 0.01). At follow-up there were significant correlations between sex and SBP (r = 0.37, P < 0.01) and left ventricular mass (r = 0.54, P < 0.001). In multivariate analyses sex emerged as a factor explaining the variance of both SBP and left ventricular mass at follow-up. In conclusion a difference in BP between male and female subjects emerges concomitantly with differences in body size during adolescence. An independent influence of sex on both BP and left ventricular mass appears in parallel with this.


High haemoglobin values during medical treatment of hypertension.
In 123 patients with arterial hypertension the haemoglobin values were determined before and during long-term antihypertensive drug treatment. The haemoglobin values found before treatment did not differ from those found in the normal population. In both sexes the haemoglobin values showed a significant increase after prolonged treatment. In males the average values rose from 15.1 to 16.7 g./100 ml., and in females from 13.96 to 14.82 g./100 ml. The increase in the haemoglobin concentration does not seem to be clearly correlated to the duration of treatment or to the decrease produced in mean blood pressure. On the other hand, the increase in haemoglobin depended to some extent on the nature of treatment. Diuretics alone resulted in a moderate increase only, whereas diuretics in combination with other antihypertensive drugs produced a more pronounced increase in haemoglobin values.


Selective Targeting of the Interconversion between Glucosylceramide and Ceramide by Scaffold Tailoring of Iminosugar Inhibitors.
A series of simple C-alkyl pyrrolidines already known as cytotoxic inhibitors of ceramide glucosylation in melanoma cells can be converted into their corresponding 6-membered analogues by means of a simple ring expansion. This study illustrated how an isomerisation from iminosugar pyrrolidine toward piperidine could invert their targeting from glucosylceramide (GlcCer) formation toward GlcCer hydrolysis. Thus, we found that the 5-membered ring derivatives did not inhibit the hydrolysis reaction of GlcCer catalysed by lysosomal β-glucocerebrosidase (GBA). On the other hand, the ring-expanded C-alkyl piperidine isomers, non-cytotoxic and inactive regarding ceramide glucosylation, revealed to be potent inhibitors of GBA. A molecular docking study showed that the positions of the piperidine ring of the compound 6b and its analogous 2-O-heptyl DIX 8 were similar to that of isofagomine. Furthermore, compound 6b promoted mutant GBA enhancements over 3-fold equivalent to that of the related O-Hept DIX 8 belonging to one of the most potent iminosugar-based pharmacological chaperone series reported to date.


Development of a High-Resolution Multi-Locus Microsatellite Typing Method for Colletotrichum gloeosporioides.
Colletotrichum gloeosporioides is an economically important fungal pathogen causing substantial yield losses indifferent host plants. To understand the genetic diversity and molecular epidemiology of this fungus, we have developed a novel, high-resolution multi-locus microsatellite typing (MLMT) method. Bioinformatic analysis of C. gloeosporioides unannotated genome sequence yielded eight potential microsatellite loci, of which five, CG1 (GT)n, CG2 (GT1)n, CG3 (TC)n, CG4 (CT)n, and CG5 (CT1)n were selected for further study based on their universal amplification potential, reproducibility, and repeat number polymorphism. The selected microsatellites were used to analyze 31 strains of C. gloeosporioides isolated from 20 different host plants from India. All microsatellite loci were found to be polymorphic, and the approximate fragment sizes of microsatellite loci CG1, CG2, CG3, CG4, and CG5 were in ranges of 213-241, 197-227, 231-265, 209-275, and 132-188, respectively. Among the 31 isolates, 55 different genotypes were identified. The Simpson's index of diversity (D) values for the individual locus ranged from 0.79 to 0.92, with the D value of all combined five microsatellite loci being 0.99. Microsatellite data analysis revealed that isolates from Ocimum sanctum, Capsicum annuum (chili pepper), and Mangifera indica (mango) formed distinct clusters, therefore exhibited some level of correlation between certain genotypes and host. The developed MLMT method would be a powerful tool for studying the genetic diversity and any possible genotype-host correlation in C. gloeosporioides.


Optical sectioning for optical scanning holography using phase-space filtering with Wigner distribution functions.
We propose a novel optical sectioning method for optical scanning holography, which is performed in phase space by using Wigner distribution functions together with the fractional Fourier transform. The principle of phase-space optical sectioning for one-dimensional signals, such as slit objects, and two-dimensional signals, such as rectangular objects, is first discussed. Computer simulation results are then presented to substantiate the proposed idea.


Riemann zeros, prime numbers, and fractal potentials.
Using two distinct inversion techniques, the local one-dimensional potentials for the Riemann zeros and prime number sequence are reconstructed. We establish that both inversion techniques, when applied to the same set of levels, lead to the same fractal potential. This provides numerical evidence that the potential obtained by inversion of a set of energy levels is unique in one dimension. We also investigate the fractal properties of the reconstructed potentials and estimate the fractal dimensions to be D=1.5 for the Riemann zeros and D=1.8 for the prime numbers. This result is somewhat surprising since the nearest-neighbor spacings of the Riemann zeros are known to be chaotically distributed, whereas the primes obey almost Poissonlike statistics. Our findings show that the fractal dimension is dependent on both level statistics and spectral rigidity, Delta(3), of the energy levels.


Moral orientation and relationships in school and adolescent pro- and antisocial behaviors: a multilevel study.
This multilevel study examined the relationships between moral climate factors and prosocial as well as antisocial behaviors inside and outside the school (school misconduct, delinquent behavior, and vandalism). The moral climate factors were punishment- and victim-based moral orientation, relationships among students, and teacher-student relationships. The analyses of data from 670 students in 69 classes showed that the classroom-level variables only had a significant impact on misconduct at school of students aged 12 to 20. For the other outcome variables, the student-level variables (student and teacher-student relationships, but especially students' moral orientation) were significant. A novel finding was that a positive teacher-student relationship not only proved to be related to less misconduct inside the school but also to less delinquent behavior and vandalism outside the school. This indicates that the teacher is an important socializing agent for adolescent behavior in general.


[The detection and investigation of traces of biological origin by cytological methods].
Thirty-four expert evaluations are analyzed in 1993-1996, in which trace particles of animal origin were pieces of material evidence. The presence of cells was certified and their species, group, and organ tissue appurtenance identified. Expert potentialities depending on the time when the object of study was formed and type of examined tissue are discussed. Combined use of biological and cytological methods permits an informative expert conclusion in cases when the traditional biological methods fail.


Increased activity of Th-17 and Th-9 lymphocytes and a skewing of the post-thymic differentiation pathway are seen in Alzheimer's disease.
Inflammatory mediators are responsible for the neuroinflammation observed in Alzheimer's disease (AD), a phenomenon that might be the culprit of disease or, possibly, a reaction to pathology. To better investigate inflammation in AD we performed an extensive immunophenotypic and functional analysis of amyloid-beta (Aβ) stimulated T lymphocytes in patients with a diagnosis of AD comparing data to those obtained in individuals with mild cognitive impairment (MCI) or aged-matched healthy individuals (HC). Results showed that IL-21- and IL-9-producing Aβ stimulated CD4(+) T cells, as well as IL-23- and IL-6-producing monocytes and CD4(+) T cells expressing the RORγ and NFATc1 transcriptional factors (TF), were significantly increased, whereas IL-10-producing monocytes were decreased in AD. Notably, GATA-3 TF-expressing CD4(+) T lymphocytes were significantly increased in MCI alone. Analysis of the post-thymic differentiation pathway indicated that Aβ specific naïve and central memory CD4(+) T lymphocytes were diminished whereas effector memory and terminally differentiated CD4(+) T lymphocytes were increased in AD and MCI compared to HC. Data herein indicate that cytokines (IL-21, IL-6, IL-23) and TF (RORγ) involved in the differentiation of Th-17 cells), as well as cytokines (IL-21, IL-22) generated by such cells, and IL-9, produced by Th-9 cells, are significantly increased in AD. This is accompanied by a shift of post-thymic differentiation pathways favoring the accumulation of differentiated, effector T lymphocytes. These data shed light on the nature of AD-associated neuroinflammation. A better understanding of the complexity of this phenomenon could facilitate the search for novel therapeutic strategies.


The geosimulation of West Nile virus propagation: a multi-agent and climate sensitive tool for risk management in public health.
Since 1999, the expansion of the West Nile virus (WNV) epizooty has led public health authorities to build and operate surveillance systems in North America. These systems are very useful to collect data, but cannot be used to forecast the probable spread of the virus in coming years. Such forecasts, if proven reliable, would permit preventive measures to be put into place at the appropriate level of expected risk and at the appropriate time. It is within this context that the Multi-Agent GeoSimulation approach has been selected to develop a system that simulates the interactions of populations of mosquitoes and birds over space and time in relation to the spread and transmission of WNV. This simulation takes place in a virtual mapping environment representing a large administrative territory (e.g. province, state) and carried out under various climate scenarios in order to simulate the effects of vector control measures such as larviciding at scales of 1/20,000 or smaller. After setting some hypotheses, a conceptual model and system architecture were developed to describe the population dynamics and interactions of mosquitoes (genus Culex) and American crows, which were chosen as the main actors in the simulation. Based on a mathematical compartment model used to simulate the population dynamics, an operational prototype was developed for the Southern part of Quebec (Canada). The system allows users to modify the parameters of the model, to select various climate and larviciding scenarios, to visualize on a digital map the progression (on a weekly or daily basis) of the infection in and around the crows' roosts and to generate graphs showing the evolution of the populations. The basic units for visualisation are municipalities. In all likelihood this system might be used to support short term decision-making related to WNV vector control measures, including the use of larvicides, according to climatic scenarios. Once fully calibrated in several real-life contexts, this promising approach opens the door to the study and management of other zoonotic diseases such as Lyme disease.


Intoxication caused by new psychostimulants: analytical methods to disclose acute and chronic use of benzofurans and ethylphenidate.
The acute and chronic toxicity of several new psychoactive substances (NPS) is unknown, and only little information is available on the pharmacology and toxicology, toxicokinetics, and detectability in body samples of such new compounds. We here propose analytical methods to disclose acute and chronic use of two types of new psychostimulants: benzofurans and ethylphenidate and we applied them to a real case of a subject attending Emergency Department with signs of acute intoxication due to psychotropic drug(s). After a urinary immunoassay screening which gave a positivity to amphetamines, general unknown gas chromatography-mass spectrometry (GC-MS) urine analysis identified 5-(2-methylaminopropyl)benzofuran (5-MAPB), 5-(2-aminopropyl)benzofuran (5-APB), 5-(2-ethylaminopropyl)benzofuran (5-EAPB), ethylphenidate, and ritalinic acid. All these substances were confirmed and quantified not only in urine but also in serum samples at different times after hospitalization by GC-MS and ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). Two subsequent 2-cm hair segments were also analyzed and tested positive for the above reported substances, evidencing repeated use. The matching quantitative results in all the analyzed biological matrices demonstrated that both analytical methodologies were suitable to correctly quantify NPS involved in the current intoxication. The objective assessment of acute and chronic intoxication by the above reported compounds demonstrate that the development of analytical methods aiming at the detection of a broad spectrum of compounds in conventional and non-conventional biological matrices is helpful when facing the new challenging threat of intoxications caused by NPS.


Comparison of oral robenacoxib and carprofen for the treatment of osteoarthritis in dogs: a randomized clinical trial.
The efficacy and tolerability of robenacoxib for the treatment of osteoarthritis in dogs were evaluated in a prospective, multicenter, randomized, noninferiority design clinical trial. A total of 32 dogs presenting with osteoarthritis were allocated randomly to receive, orally once daily for 28 days, either 1-2 mg/kg robenacoxib (n=21) or 3.5-5 mg/kg carprofen (n=11). Dogs were assessed by clinicians and owners using numerical rating scale scores at baseline and days 14 and 28. The primary efficacy endpoint was the global functional disability score, which was the sum of clinician scores for standing posture, lameness at walk, lameness at trot, willingness to raise the contralateral limb and pain at palpation. There was a good to excellent level of efficacy in both treatment groups. Differences between days 14 and 28 compared to day 0 were significant for all 11 clinician and owner scores for robenacoxib, and for 6 of 11 scores for carprofen. The efficacy of robenacoxib was numerically superior to carprofen for all 13 endpoints, but differences were not statistically significant. For the global functional disability score, the estimated efficacy of robenacoxib was 1.244 (95% confidence interval 0.555-2.493) relative to carprofen. The tolerability of both treatments was good as assessed from adverse events, clinical signs, and hematology and serum biochemistry variables. In conclusion, once daily administration of robenacoxib tablets had noninferior efficacy and tolerability compared to carprofen for the treatment of the clinical signs of osteoarthritis in dogs.


Factors Influencing Time to Adoption for Dogs in a Provincial Shelter System in Canada.
Millions of companion animals are relinquished to shelters each year. For each dog entering, their characteristics and the characteristics of the shelter holding him/her influence their time to adoption. Using a Cox proportional hazards frailty model, these issues were explored using data from 31 shelters within the British Columbia Society for the Prevention of Cruelty to Animals shelter system. Results indicated that surrender reason, age, original source, coat color, breed, human population density of the shelter location, and year the nonhuman animal entered the shelter significantly influenced time to adoption. This study identified characteristics of dogs that make them less likely to be adopted quickly. Given limited resources, these results may help inform shelters on how to best allocate resources, particularly in interventions to improve adoption rates in shelters.


Insulin resistance is an important risk factor for cognitive impairment in elderly patients with primary hypertension.
Insulin resistance plays a role in the development of dementia and hypertension. We investigated a possible relationship between cognitive impairment and insulin resistance in elderly Chinese patients with primary hypertension. One hundred and thirty-two hypertensive elderly patients (>60 years) were enrolled in this study, and assigned into either the cognitive impairment group (n=61) or the normal cognitive group (n=71). Gender, age, education, body mass index (BMI), waist hip ratio (WHR), total cholesterol (TC), triglyceride (TG), C-reactive protein (CRP), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), creatinine (Cr), fasting plasma glucose (FPG), fasting insulin (FINS), homeostasis model of assessment for insulin resistance index (HOMA-IR), systolic blood pressure, diastolic blood pressure, smoking history, atherosclerosis and the proportion of uncontrolled hypertension were compared between the two groups. Multi-factorial logistic regression analysis was performed. No significant differences were found in gender, age, TC, CRP, HDL-C, LDL-C, Cr, BP, smoking history, atherosclerosis and the proportion of uncontrolled hypertension between the two groups. The cognitive impairment group had lower education levels, and higher BMI, WHR, TG, FPG, FINS, and HOMA-IR levels than the control group. Logistic regression analysis revealed the levels of education, BMI, WHR, and HOMA-IR as independent factors that predict cognitive impairment in patients. Our study demonstrates that poor education and increased BMI, WHR, and HOMA-IR are independent risk factors for cognitive impairment in elderly patients with hypertension. Insulin resistance plays an important role in the development of cognitive impairment in primary elderly hypertensive patients.


Individual normal laboratory values. Preliminary observations.
A brief and preliminary outline is given describing a consecutive and continuing study of laboratory blood values of only 12 of 48 "normal healthy" subjects with only a few values given. The major emphasis is that of obtaining blood from each subject over a 12-week period to be repeated annually in order to determine individual values and in obtaining a chemical identification of each subject with the anticipation that more information will be available concerning the meaning and limiting parameters of "normal" biologic values. Such a study is made available through the application of modern advances in automation and the wide use of computers. It seems likely that some disorders can be discovered before clinically apparent, with the hope that consequent preventive measures and therapy may be more effective. The few values presented represent only a small part of those yet to be obtained. Much more work is needed in this study of normal values whose parameters must be further defined.


Endovascular treatment of vein of Galen dural arteriovenous fistula presenting as dementia.
Dural arteriovenous fistula (DAVF) is an important cause of neurological dysfunction that is often misdiagnosed, especially in elderly population. Galenic DAVFs are a subtype of the rare falcotentorial DAVFs with a high risk of hemorrhage and aggressive clinical course. In most cases, DAVFs present with pulsatile tinnitus, headache, or orbital symptoms such as chemosis and proptosis. We report a patient with DAVF of Vein of Galen presented with progressive dementia, treated by Onyx embolisation and had good clinical outcome.


[Electron microscopic studies on the glycocalyx of the utricular macula following amikacin in guinea pigs].
Twelve guinea pigs were randomly separated into 4 groups, one group as control, the other groups were given different dosages of Amikacin for 5 days. Ten days later, the animals were killed by decapitation and the right utricular maculae removed. After staining with reuthenium red, the glycocalyx was observed under TEM. In the control group, the glycocalyx was found covering the surface of both cells and sensory hairs and forming cross links with all the sensory hairs in the same hair band. The main changes in the experimental groups were: 1. decrease or disappearance of glycocalyx on the surface of cells and hairs; 2. fusion of sensory hairs and large blebs formed at the side of hairs. The results of the present study suggest that the decrease or disappearance of glycocalyx may be the first change of the damages caused by Amikacin.


Insights into the activity of maturation inhibitor PF-46396 on HIV-1 clade C.
HIV maturation inhibitors are an emerging class of anti-retroviral compounds that inhibit the viral protease-mediated cleavage of the Gag, CA-SP1 (capsid-spacer peptide 1) peptide to mature CA. The first-in-class maturation inhibitor bevirimat (BVM) displayed potent activity against HIV-1 clade B but was ineffective against other HIV-1 clades including clade C. Another pyridone-based maturation inhibitor, PF-46396 displayed potent activity against HIV-1 clade B. In this study, we aimed at determining the activity of PF-46396 against HIV-1 clade C. We employed various biochemical and virological assays to demonstrate that PF-46396 is effective against HIV-1 clade C. We observed a dose dependent accumulation of CA-SP1 intermediate in presence of the compound. We carried out mutagenesis in the CA- SP1 region of HIV-1 clade C Gag and observed that the mutations conferred resistance against the compound. Many mutations inhibited Gag processing thereby reducing virus release in the absence of the compound. However, presence of PF-46396 rescued these defects and enhanced virus release, replication capacity and infectivity of HIV-1 clade C. These results put together identify PF-46396 as a broadly active maturation inhibitor against HIV-1 clade B and C and help in rational designing of novel analogs with reduced toxicity and increased efficacy for its potential use in clinics.


Gender differences in patients undergoing TAVI: a multicentre study.
In patients undergoing surgical valve replacement for severe aortic stenosis, female gender was associated with worse outcomes, not persisting after multivariable adjustment for baseline clinical differences, while contrasting data are reported about TAVI. From January 2007 to December 2011 all patients with severe symptomatic aortic stenosis undergoing TAVI at our institutions were included in the present study, and were divided into two cohorts according to their gender. All endpoints were adjudicated according to VARC definition. Three hundred and seventy-seven patients were included: 161 male and 216 female. Male patients reported higher rates of previous coronary revascularisation, while both ejection fraction and mean aortic gradient were higher in female patients. At 30 days, rates of cardiovascular death were similar (6.0% vs. 8.1%; p=0.793), while overall bleedings (44% vs. 25%; p=0.024) and life-threatening bleedings (21.1% vs. 12.7%, p=0.016) were higher in female patients, also after multivariate analysis (OR 3.44; 1.23-9.22, and OR 2.1; 1.1-4.0, CI: 95%, respectively). Major vascular complications showed a tendency to be higher in female patients (12.9% vs. 9.8%, p=0.449). At a mean follow-up of 490 ± 250 days, no significant difference was reported between men and women for all endpoints, and after multivariate adjustment only life-threatening bleeding was reported as a predictor of death (OR 8.2:3.8-17, CI: 95%). TAVI can be an effective and safe strategy in high surgical risk patients, regardless of the gender; life-threatening bleedings were reported more frequently in female patients, being the only independent predictor of death.


The PROCESS experiment: amino and carboxylic acids under Mars-like surface UV radiation conditions in low-earth orbit.
The search for organic molecules at the surface of Mars is a top priority of the next Mars exploration space missions: Mars Science Laboratory (NASA) and ExoMars (ESA). The detection of organic matter could provide information about the presence of a prebiotic chemistry or even biological activity on this planet. Therefore, a key step in interpretation of future data collected by these missions is to understand the preservation of organic matter in the martian environment. Several laboratory experiments have been devoted to quantifying and qualifying the evolution of organic molecules under simulated environmental conditions of Mars. However, these laboratory simulations are limited, and one major constraint is the reproduction of the UV spectrum that reaches the surface of Mars. As part of the PROCESS experiment of the European EXPOSE-E mission on board the International Space Station, a study was performed on the photodegradation of organics under filtered extraterrestrial solar electromagnetic radiation that mimics Mars-like surface UV radiation conditions. Glycine, serine, phthalic acid, phthalic acid in the presence of a mineral phase, and mellitic acid were exposed to these conditions for 1.5 years, and their evolution was determined by Fourier transform infrared spectroscopy after their retrieval. The results were compared with data from laboratory experiments. A 1.5-year exposure to Mars-like surface UV radiation conditions in space resulted in complete degradation of the organic compounds. Half-lives between 50 and 150 h for martian surface conditions were calculated from both laboratory and low-Earth orbit experiments. The results highlight that none of those organics are stable under low-Earth orbit solar UV radiation conditions.


Chlorophyll-a derivatives with various hydrocarbon ester groups for efficient dye-sensitized solar cells: static and ultrafast evaluations on electron injection and charge collection processes.
Five chlorophyll-a derivatives, chlorins-1-5 possessing C3(2)-carboxy and O17(4)-esterified hydrocarbon groups including methyl, hexyl, dodecyl, 2-butyloctyl, and cholesteryl were synthesized. Their performance as sensitizers in dye-sensitized solar cells (DSSCs) was compared. These sensitizers have similar surface coverage on the unit surface of TiO(2) film and their absorption spectra on transparent TiO(2) films were identical. On the basis of DFT and TD-DFT calculations of these sensitizers in ethanol, a major difference between them was the geometry of the hydrocarbon ester group, to affect their electron injection and charge recombination with the TiO(2) electrode rather than the energy level of their molecular orbitals. DSSC based on chlorin-3 with a dodecyl ester group gave a solar energy-to-electricity conversion efficiency of 8%, which was the highest among all the chlorophyllous sensitizers. The large photocurrent in the chlorin-3 sensitized solar cell can be explained by the least impedance in the electrolyte-dye-TiO(2) interface in electrical impedance spectroscopy measurements. Subpicosecond time-resolved absorption spectroscopic studies have also been carried out to evaluate the electron injection and charge recombination dynamics in the dye-TiO(2) interface. For the electron injection and charge recombination processes, a charge separated state of the dye-TiO(2) complex has been found to be free from the type and concentration of dye sensitizer, reflecting the same type of electron transfer process for all the five chlorin sensitizers. A new quenching pathway of the dye excitation, which is probably from the exciton annihilation, in addition of the charge recombination has been observed for chlorin-1 and chlorin-5, but not for chlorin-3. The higher open-circuit photocurrent observed in the present dyes with larger ester groups can be attributed to the reduced leaking of charges in the TiO(2)-electrolyte interface, which was supported by the longer electron lifetimes.


Invasive early-onset neonatal group B streptococcal cases--Alaska, 2000-2004.
We conducted a review of invasive early-onset neonatal group B Streptococcus (GBS) infections that occurred during 2000-2004 in Alaska to determine the proportion of cases that might have been prevented by complete implementation of the 2002 Centers for Disease Control and Prevention (CDC) guidelines. Cases were identified from statewide laboratory-based surveillance conducted by the CDC Arctic Investigations Program, and from the Alaska Medicaid database using International Classification of Diseases 9 codes 038.0, 041.02, 320.2, and 482.3. Neonates were considered to have early-onset disease if clinical illness within 6 days after birth was accompanied by GBS isolation from a normally sterile site. Maternal and neonatal medical records were reviewed. Potentially preventable cases were those for whom the 2002 CDC GBS maternal screening and intrapartum antimicrobial prophylaxis (IAP) guidelines were not completely implemented. Preventability of events not related to clinician implementation of the guidelines were not considered. Twenty-one neonates with invasive early-onset GBS disease were identified (0.42/1,000 live births). Three of the eight mothers for whom IAP was indicated, did not receive adequate IAP. Nine of the 13 mothers for whom there was no indication for IAP, had not been screened appropriately. Therefore, a total of 12 neonates were determined to have had potentially preventable GBS disease. Over 50% of the invasive early-onset neonatal GBS cases in Alaska were potentially preventable. The majority of these cases may have been prevented by closer adherence to either specific IAP administration guidelines or to maternal screening guidelines.


[Relation between pure tone audiometry and speech audiometry in various hearing-impaired listeners].
Pure tone audiometry and speech audiometry were performed in 30 normal-hearing listeners and 75 listeners suffering from conductive, sensorineural hearing loss or presbyacusia. The results were as follows: (1) For listeners with conductive or mild sensorineural hearing loss, or with mild presbyacusia, the difference between speech reception thresholds (SRT) and pure tone averages (PTA) was not significant (P > 0.05). (2) For listeners with profound sensorineural hearing loss or presbyacusia, the SRT were about 10 dB greater than the PTA. (3) The maximum discrimination score (PBXax) measurements should be made at a comfortable intensity for each listener, some 30 dB above the SRT in general. We conclude that the PTA can replace the SRT directly for the listeners described in (1) or that the PTA, by adding 10 dB, can also be used as a replacement of the SRT for the listeners described in (2).


GDE2-Dependent Activation of Canonical Wnt Signaling in Neurons Regulates Oligodendrocyte Maturation.
Neurons and oligodendrocytes communicate to regulate oligodendrocyte development and ensure appropriate axonal myelination. Here, we show that Glycerophosphodiester phosphodiesterase 2 (GDE2) signaling underlies a neuronal pathway that promotes oligodendrocyte maturation through the release of soluble neuronally derived factors. Mice lacking global or neuronal GDE2 expression have reduced mature oligodendrocytes and myelin proteins but retain normal numbers of oligodendrocyte precursor cells (OPCs). Wild-type (WT) OPCs cultured in conditioned medium (CM) from Gde2-null (Gde2KO) neurons exhibit delayed maturation, recapitulating in vivo phenotypes. Gde2KO neurons show robust reduction in canonical Wnt signaling, and genetic activation of Wnt signaling in Gde2KO neurons rescues in vivo and in vitro oligodendrocyte maturation. Phosphacan, a known stimulant of oligodendrocyte maturation, is reduced in CM from Gde2KO neurons but is restored when Wnt signaling is activated. These studies identify GDE2 control of Wnt signaling as a neuronal pathway that signals to oligodendroglia to promote oligodendrocyte maturation.


Defining the volume dependence of multiple K flux pathways of trout red blood cells.
The volume sensitivity of different K flux pathways has been determined in trout red blood cells subjected to volume perturbation. Gentle hyposmotic swelling induced a K influx in a Cl-containing saline but not in NO3- or methanesulfonate (MeSF)-containing salines, consistent with the activation of a Cl-dependent flux. Extreme hyposmotic swelling led to larger K fluxes in all salines but with reduced anion discrimination of the Cl-dependent flux. In contrast to these graded responses, isosmotic swelling using ammonium chloride or beta-adrenergic stimulation activated only Cl-dependent fluxes in an all-or-none fashion. The relationship between the hyposmotically and isosmotically induced pathways was studied by coactivation using either ammonium chloride or isoproterenol with anisosmotic treatment. Cells in ammonium chloride-containing hyposmotic salines showed no additive K flux over that induced by hyposmotic treatment alone, indicating that the isosmotically induced Cl-dependent flux was identical to the hyposmotically induced Cl-dependent flux. However, cells coactivated by hyposmotic and beta-adrenergic treatment showed a small Cl-dependent flux in addition to that induced by hyposmotic treatment alone. This small third component was unaffected by anisosmotic treatment. We conclude that the major Cl-dependent and Cl-independent K flux pathways are distinct and separate and that the former has an anion dependence that varies with cell volume and a volume sensitivity that varies with ionic strength.


The in(put)s and out(put)s of the Drosophila circadian clock.
Circadian clocks are aligned to the 24-h environmental day via zeitgebers, and information from the clock must be relayed to the rest of the organism. Recent studies addressing temperature as an input to, and the generation of rhythmic gene expression as an output from, the Drosophila central clock are discussed.


Recent Updates on Obesity Treatments: Available Drugs and Future Directions.
In the last thirty years, obesity has reached epidemic proportions and is now regarded as a major health issue in contemporary society trending to serious economic and social burdens. The latest projections of the World Health Organization are alarming. By 2030, nearly 60% of the worldwide population could be either obese or overweight, highlighting the needs to find innovative treatments. Currently, bariatric surgery is the most effective way to efficiently lower body mass. Although great improvements in terms of recovery and patient care were made in these surgical procedures, bariatric surgery remains an option for extreme forms of obesity and seems unable to tackle obesity pandemic expansion. Throughout the last century, numerous pharmacological strategies targeting either peripheral or central components of the energy balance regulatory system were designed to reduce body mass, some of them reaching sufficient levels of efficiency and safety. Nevertheless, obesity drug therapy remains quite limited on its effectiveness to actually overcome the obesogenic environment. Thus, innovative unimolecular polypharmacology strategies, able to simultaneously target multiple actors involved in the obesity initiation and expansion, were developed during the last ten years opening a new promising avenue in the pharmacological management of obesity. In this review, we first describe the clinical features of obesity-associated conditions and then focus on the outcomes of currently approved drug therapies for obesity as well as new ones expecting to reach the clinic in the near future.


Albuminuria Predicts Short-Term Worsening Renal Function After Transcatheter Aortic Valve Replacement.
Albuminuria is a major risk factor of cardiovascular events, however, the impact of albuminuria on clinical outcomes of patients undergoing transcatheter aortic valve replacement (TAVR) has not been fully investigated. This retrospective study included 206 patients who underwent TAVR for severe aortic stenosis. Patients were divided into two groups according to the preoperative urinary albumin-to-creatinine ratio (ACR): high (ACR ≥ 30 mg/g) and low (ACR < 30 mg/g). The incidence of 1-month worsening renal function (WRF), defined as a decrease in estimated glomerular filtration rate (eGFR) ≥10% from baseline after TAVR, was investigated. Patients with high ACR had acute kidney injury (8.5% vs. 1.0%, p = 0.01) and 1-month WRF (29.2% vs. 12.0%, p = 0.002) more frequently than those with low ACR. High ACR was independently associated with 1-month WRF (odds ratio, 3.72; 95% confidence interval, 1.72-8.08; p < 0.001). Albuminuria can be a useful predictor of deterioration of renal function at various time points after TAVR.


Cavernous nerve stimulation through an extrapelvic subpubic approach: role in penile erection.
The results of cavernous nerve (CN) stimulation through an extrapelvic subpubic approach in 12 dogs are presented. The nerve was exposed and identified by a magnifying loupe and electric stimulation. A bioplar platinum electrode was then applied to the nerve and a radiofrequency receiver implanted subcutaneously into the abdomen. The CN was stimulated (pulse width 200 microseconds, range density 2-6 microCi/cm2/phase) and the intracavernous pressure was recorded. CN stimulation led to intracavernous pressure increase. With increasing stimulus frequency, the pressure increased while the latency and stimulation phase decreased, until at 60 Hz frequency no further changes occurred. Increasing CN stimulation was accompanied by a progressive increase in penile length and diameter leading to full erection at a frequency of 60 Hz. The penile response was more rapid and of a shorter duration with stronger stimulation. CN stimulation could be repeated indefinitely without fatigue after an off-time of double the period of the stimulation phase. The extrapelvic subpubic approach to CN is simple and without complications, and can be applied safely in humans.


A case of perifollicular fibroma.
Perifollicular fibroma (PFF) is a very rare proliferative lesion of the perifollicular sheath. A flesh-colored, dome-shaped papulonodule is usually located on the face or the neck. These papulonodules may be single or multiple. Histologically, the lesion consists of a concentric arrangement of cellular fibrous tissue around a normal hair follicle. We describe here an unusual case of PFF in a 41-year-old female who had an asymptomatic small skin colored papule on the scalp, and this is an uncommon location for PFF.


Ray helicity: geometric invariant for multidimensional resonant wave conversion.
For a multicomponent wave field propagating into a multidimensional conversion region, the rays are shown to be helical, in general. For a ray-based quantity to have a fundamental physical meaning, it must be invariant under the following two groups of transformations, which are used to construct solutions: congruence transformations (which involve linear combinations of components of the multicomponent wave field) and canonical transformations (which act on the ray phase space). It is shown that for conversion between two waves there is a new invariant not previously discussed: the intrinsic helicity of the ray.


Ultrasound evaluation of bone in sport: the role of physical activity in young volleyball females players.
Physical activity represents an effective incentive to achieve and maintain the peak bone mass. Our study has been evaluating the heel ultra-sound parameters in a group of female volleyball players. We found some interesting results as follows: BMD estimated, 0.675±0.144 g/cm2; QUI 120.2±20.9; SOS 1590.4±32.3 m/S; BUA 95.5±20.1 dB/ MHz. Each parameter observed on the volleyball players group has resulted significantly higher (p<0.05-0.01) compared to the same parameters within the control group. Our work confirms that an aerobic physical activity represents an effective mechanical stimulus to the bone, allowing of observing an index of bone quality significantly higher compared to the people who does not do any regular physical activity.


Oxidative stress during coronary artery bypass operations: importance of surgical trauma and drug treatment.
To investigate oxidative stress and myocardial injury at different stages of coronary artery bypass grafting (CABG). Twenty patients underwent CABG with use of cardiopulmonary bypass (CPB) and with intermittent sampling of plasma and urine. Main markers were: 8-iso-PGF2alpha (oxidative stress); troponin T (myocardial injury); and 15-keto-dihydro-PGF2alpha and hsCRP (inflammation). Plasma 8-iso-PGF2alpha increased after start of surgery, but there was no further rise during CPB or after aortic cross-clamp release and no significant myocardial arterio-venous differences. An increase in troponin T was seen early after the operation, but no relationship was established between 8-iso-PGF2alpha and troponin T. 8-iso-PGF2alpha levels were elevated by preoperative withdrawal of acetylsalicylic acid (ASA) but reduced by intraoperative use of heparin. 15-keto-dihydro-PGF2alpha was elevated during operation and hsCRP following operation. In the present study oxidative stress was multifactorial in origin with main impacts from surgical trauma, less from CPB and little if any from myocardial ischemia-reperfusion events. In addition, cardiovascular drugs in common use like ASA and heparin seemed to influence the pro- and antioxidant balance, a finding that has to be confirmed in future studies.


Methionine synthase is localized to the nucleus in Pichia pastoris and Candida albicans and to the cytoplasm in Saccharomyces cerevisiae.
Methionine synthase (MS) catalyzes methylation of homocysteine, the last step in the biosynthesis of methionine, which is essential for the regeneration of tetrahydrofolate and biosynthesis of S-adenosylmethionine. Here, we report that MS is localized to the nucleus of Pichia pastoris and Candida albicans but is cytoplasmic in Saccharomyces cerevisiae The P. pastoris strain carrying a deletion of the MET6 gene encoding MS (Ppmet6) exhibits methionine as well as adenine auxotrophy indicating that MS is required for methionine as well as adenine biosynthesis. Nuclear localization of P. pastoris MS (PpMS) was abrogated by the deletion of 107 C-terminal amino acids or the R742A mutation. In silico analysis of the PpMS structure indicated that PpMS may exist in a dimer-like configuration in which Arg-742 of a monomer forms a salt bridge with Asp-113 of another monomer. Biochemical studies indicate that R742A as well as D113R mutations abrogate nuclear localization of PpMS and its ability to reverse methionine auxotrophy of Ppmet6 Thus, association of two PpMS monomers through the interaction of Arg-742 and Asp-113 is essential for catalytic activity and nuclear localization. When PpMS is targeted to the cytoplasm employing a heterologous nuclear export signal, it is expressed at very low levels and is unable to reverse methionine and adenine auxotrophy of Ppmet6 Thus, nuclear localization is essential for the stability and function of MS in P. pastoris. We conclude that nuclear localization of MS is a unique feature of respiratory yeasts such as P. pastoris and C. albicans, and it may have novel moonlighting functions in the nucleus.


A modified prosthesis eversion technique for proximal anastomosis in ascending aorta replacement.
One of the crucial aspects of ascending aorta replacement is to achieve hemostasis of the proximal anastomosis. This study aimed to describe a modified prosthesis eversion technique for proximal anastomosis in ascending aorta replacement and compare its operative outcomes with the conventional prosthesis eversion technique. We conducted a retrospective analysis of all consecutive patients who had ascending aortic aneurysm and underwent ascending aorta replacement with the modified or conventional prosthesis eversion technique between January 2019 and December 2022 in our center. A total of 108 patients were included: 55 in the modified group and 53 in the conventional group. The durations of cardiopulmonary bypass, aortic cross-clamping and total operation in the conventional group were longer than those in the modified group. Furthermore, perioperative blood loss and the incidence of re-exploration for bleeding were significantly lower in the modified group. Accordingly, patients in the conventional group accepted more blood transfusion. The modified group had a shorter duration in intensive care unit (ICU) and hospital, and lower total hospitalization costs than those in the conventional group. The modified prosthesis eversion technique is an effective alternative for proximal anastomosis in ascending aorta replacement, with less blood loss, shorter operation time, and lower rate of postoperative complications compared with the conventional technique.


Endovascular techniques for placement of long-term chemotherapy catheters.
To analyze the results from using endovascular techniques to place long-term chemotherapy catheters when advancing the catheter using the external jugular vein is difficult due to obstructions or kinking. Between July 1997 and August 2000, 320 long-term chemotherapy catheters were placed, and in 220 cases the external jugular vein was used as the primary venous approach. In 18 of these patients, correct positioning was not achieved and several endovascular techniques were then utilized to overcome these obstacles, including manipulation of a J-wire with a moveable core, venography, and the exchange wire technique. In 94.5% of the patients with difficulties in obtaining the correct positioning, we were able to advance the long-term catheter to the desired position with the assistance of endovascular techniques. Venography and endovascular guidance techniques are useful for the placement of long-term catheters in the external jugular vein.


In-vitro susceptibility testing by agar dilution method to determine the minimum inhibitory concentrations of amphotericin B, fluconazole and ketoconazole against ocular fungal isolates.
To standardize in-vitro antifungal susceptibility testing by agar dilution method to find out the minimum inhibitory concentration (MIC) of amphotericin B, fluconazole and ketoconazole on ocular fungal isolates. A total of 180 ocular fungal isolates (130 filamentous fungi and 50 yeasts) were included. The antifungal drugs such as amphotericin B (0.0625-8 microg/mL), fluconazole (0.2-819.6 microg/mL) and ketoconazole (0.025-6.4 microg/mL) were incorporated in doubling dilutions in the yeast nitrogen base medium. The MIC was determined as the lowest concentration of the antifungal drug preventing growth of macroscopically visible colonies on drug containing plates when there was visible growth on the drug-free control plates. All 50 ocular isolates of yeast were susceptible to amphotericin B, while two (4%) and five (10%) strains were resistant to fluconazole and ketoconazole respectively. Of the 130 filamentous fungi tested, six (4.6%) were resistant to amphotericin B, 49 (37.7%) and 10 (7.6%) were resistant to fluconazole and ketoconazole respectively. Percentile 50 (MIC 50) and Percentile 90 (MIC 90) for all the three antifungal agents were calculated. Aspergillus niger, Aspergillus terreus and Candida krusei were found to be resistant to fluconazole and ketoconazole. This technique was found to be reliable, cost effective and easy to perform with consistent results.


Evaluation of the role of injection therapy for benign prostatic hypertrophy.
Intraprostatic phenol injection treatment was given to 13 cases of benign prostatic hypertrophy (BPH) without urinary retention. Symptomatic improvement was significant, although objective assessment showed neither significant alteration in the size of the prostate nor in the urinary flow rate. Injection therapy is considered inadequate in the relief of prostatic obstruction.


Biodistribution and PDT efficacy of a ketochlorin photosensitizer as a function of the delivery vehicle.
C8KC is a new ketochlorin photosensitizer that must be formulated with an emulsifier because of its poor water solubility. In this report, we compare properties of Cremophor EL (CRM) and Tween 80 as delivery vehicles for C8KC. Unlike Tween 80, CRM altered the physical properties of both human and mouse plasma lipoproteins, resulting in decreased electrophoretic mobility of the individual lipoproteins along with the formation of a lipoprotein degradation product: a phospholipid fraction of low buoyant density. In human plasma, where there was sufficient low-density lipoprotein (LDL) for a distinction to be made, CRM caused a shift in binding of a ketochlorin from albumin to LDL and the degraded lipoprotein fraction. In mice bearing the RIF tumor, the use of CRM for drug formulation was associated with longer plasma and tissue persistence of C8KC, and enhanced photodynamic therapy (PDT) efficacy. These results indicate the importance of both sensitizer and vehicle as determinants of PDT efficacy.