A Universal Labeling Strategy for Nucleic Acids in Expansion Microscopy.
Expansion microscopy (ExM) enables the nanoscale imaging of ribonucleic acids (RNAs) on a conventional fluorescence microscope, providing information on the intricate patterns of gene expression at (sub)cellular resolution and within spatial context. To extend the use of such strategies, we examined a series of multivalent reagents that allow the labeling and grafting of deoxyribonucleic acid (DNA) oligonucleotide probes in a unified approach. We show that the reagents are directly compatible with third-generation in situ hybridization chain reaction RNA FISH (fluorescence in situ hybridization) techniques while displaying complete retention of the targeted transcripts. Furthermore, we validate and demonstrate that our labeling method is compatible with multicolor staining. Through oligonucleotide-conjugated antibodies, we demonstrate excellent performance in ×4 ExM and ×10 ExM, achieving a resolution of ∼50 nm in ×10 ExM for both pre- and postexpansion labeling strategies. Our results indicate that our multivalent molecules enable the rapid functionalization of DNA oligonucleotides for ExM.


Ultrasound-assisted Extraction of Ursolic Acid from the Flowers of Ixora coccinia Linn (Rubiaceae) and Antiproliferative Activity of Ursolic Acid and Synthesized Derivatives.
Ixora coccinea Linn (Rubiaceae) is an evergreen shrub with bright scarlet colored flowers found in several tropical and subtropical countries. It is used as an ornamental and medicinal plant. Phytochemical studies revealed that its major special metabolites are triterpene acids, such as ursolic and oleanolic acid. To evaluate the isolation of ursolic acid (UA) (1) from methanol extracts of I. coccinea flowers through two methodologies, to prepare four derivatives, and to evaluate the cytotoxic effect against six cancer cell lines. The UA was isolated from vegetal material by percolation at room temperature and by ultrasound-assisted extraction. The preparation of derivatives was performed according to literature methods, and the cytotoxic effects were evaluated using the MTT (3,4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide) assay. The most efficient extraction was achieved through ultrasound irradiation with a yield of 35% after KOH-impregnated silica in chromatography column. Furthermore, four derivatives (3, 5, 6, 7) of UA were prepared and evaluated, including 1, against two lung cancer (A549 and H460) and four leukemia (K562, Lucena, HL60, and Jurkat) cell lines. Generally, results showed that 1 and 7 were the most active compounds against the assayed cell lines. Also, the cytotoxic effects observed on terpenes 1 and 7 were higher when compared with cisplatin, used as positive control, with the exception of Jurkat cell line. The efficiency of such an alternative extraction method led to the principal and abundant active component, 1, of I. coccinea, thus representing a considerable contribution for promising triterpenoid in cancer chemotherapy. The ultrasound-assisted extraction of Ixora coccinea flowers improved of the ursolic acid isolationMethanolic extract from flowers of I. coccinea provided, by ultrasound irradiation, after KOH-impregnated silica in chromatography column, the ursolic acid in 35% yieldThe ursolic acid and four derivatives were prepared and assayed against two lung cancer and four leukaemia cell linesThe ursolic acid and their 3-oxo-derivative, in general, were more cytotoxic when compared to cisplatin, used as positive control Abbreviations used: MTT: 3,4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide, RP: reverse phase, TLC: thin layer chromatography, KOH: potassium hydroxide, IR: infrared, DMF: dimethylformamide, DMSO: dimethyl sulfoxide, TEA: triethylamine, RT: room temperature, EtOAc: ethyl acetate, MeOH: methanol, i-PrOH: iso-propanol, NMR: nuclear magnetic resonance, MDR: multiple drug resistance, RPMI: Roswell Park Memorial Institute.


Predicting obsessive-compulsive disorder episodes in adolescents using a wearable biosensor-A wrist angel feasibility study.
Obsessive-compulsive disorders (OCD) are marked by distress, negative emotions, mental processes and behaviors that are reflected in physiological signals such as heart rate, electrodermal activity, and skin temperature. Continuous monitoring of physiological signals associated with OCD symptoms may make measures of OCD more objective and facilitate close monitoring of prodromal symptoms, treatment progress and risk of relapse. Thus, we explored the feasibility of capturing OCD events in the real world using an unobtrusive wrist worn biosensor and machine learning models. Nine adolescents (ages 10-17 years) with mild to moderate-severe OCD were recruited from child and adolescent mental health services. Participants were asked to wear the biosensor in the lab during conditions of rest and exposure to OCD symptom-triggering stimuli and for up to 8 weeks in their everyday lives and register OCD events. We explored the relationships among physiological data, registered OCD events, age, OCD symptom severity and symptom types. In the machine learning models, we considered detection of OCD events as a binary classification problem. A nested cross-validation strategy with either random 10-folds, leave-one-subject-out, or leave-week(s)-out in both layers was used. We compared the performance of four models: logistic regression, random forest (RF), feedforward neural networks, and mixed-effect random forest (MERF). To explore the ability of the models to detect OCD events in new patients, we assessed the performance of participant-based generalized models. To explore the ability of models to detect OCD events in future, unseen data from the same patients, we compared the performance of temporal generalized models trained on multiple patients with personalized models trained on single patients. Eight of the nine participants collected biosensor signals totaling 2, 405 h and registered 1, 639 OCD events. Better performance was obtained when generalizing across time compared to across patients. Generalized temporal models trained on multiple patients were found to perform better than personalized models trained on single patients. RF and MERF models outperformed the other models in terms of accuracy in all cross-validation strategies, reaching 70% accuracy in random and participant cross-validation. Our pilot results suggest that it is possible to detect OCD episodes in the everyday lives of adolescents using physiological signals captured with a wearable biosensor. Large scale studies are needed to train and test models capable of detecting and predicting episodes. ClinicalTrials.gov: NCT05064527, registered October 1, 2021.


[Study on optimal harvest period of Lonicera Flos (Lonicera macranthoides)].
To ascertain the optimal harvest period of Lonicera Flos (Lonicera macranthoides) the configuration yield and quality of L. macranthiodes bloom verity and bud verity flower at different develop periods were Observed. The quality of L. macranthiodes which harvested at different times of the day was Compared. The configuration was significant difference between different develop period of L. macranthiodes flower. As bud growth, yield increased. Bloom verity of L. macranthoides chlorogenic acid content was significantly lower after opening (silver flower stage, golden flower stage), before opening (young bud stage, green-white stage) have no significant difference of the quality. Bud verity of L. macranthoides macranthoidin B is significant lower at yellow-white stage, young bud stage and green-white stage have no significant difference of the quality. The chlorogenic acid and isochlorogenic acid A content is significant difference between L. macranthoides harvested at different time of the day. The optimal harvest period of bloom verity is the white stage, picking time for 10:00 before and after 18:00. The optimal harvest period is the green-white stage, picking time is 8:00 before and after 18:00.


Epidemiology of rheumatoid arthritis, juvenile idiopathic arthritis and gout in two regions of the Czech Republic in a descriptive population-based survey in 2002-2003.
To estimate the annual incidence and prevalence of rheumatoid arthritis (RA), juvenile arthritis (JIA) and gout in a population based study in two regions of the Czech Republic with total population of 186,000 inhabitants. The study was conducted in the Town of Ceske Budejovice and district of Cheb in the Czech Republic (with a total population of 186,000 inhabitants) in the years 2002 and 2003. Incident cases were registered on condition that the definite diagnosis was confirmed according to existing classification criteria during the study period. Prevalence was studied on the basis of identification of established diagnosis from registers of patients of participating rheumatologists and other specialists. They were asked to report all living patients who had been diagnosed before 1st March 2002. Patients were only included in the study if their permanent address was in the selected study area. Overall, we found 48 incident and 947 prevalent cases of RA among adults (16+ years), 4 incident and 43 prevalent cases of JIA among children (less than 16 years old), and 64 incident and 425 prevalent cases of gout among adults (16+ years). The total annual incidence of RA was 31/100,000 in the adult population aged 16 years and more (95% CI 20 to 42/100,000). The prevalence of RA was 610/100,000 (95% CI 561 to 658/100,000) in the adult population. An annual incidence of gout in adults was 41/100,000 (95% CI 28 to 53/100,000). The prevalence of gout was 300/100,000 (95% CI 266 to 334/100,000). The annual incidence of JIA was 13/100,000 in children less than 16 years old (95%CI 1 to 20/100,000). The prevalence of JIA in children was 140/100,000 (95% CI 117 to 280/100,000). This study estimates the annual incidence and prevalence rates of RA, gout and JIA in the first population-based survey in the Czech Republic. The rates of RA and JIA compare well with figures reported from other countries; figures in gout seem to be lower than reported elsewhere.


Cognitive Function of Children and Adolescents with Attention-Deficit/Hyperactivity Disorder in a 2-Year Open-Label Study of Lisdexamfetamine Dimesylate.
SPD489-404 was the first 2-year safety study of lisdexamfetamine dimesylate in the treatment of attention-deficit/hyperactivity disorder in children and adolescents. In accordance with advice from the European Medicines Agency, assessment of cognitive function was a predefined safety outcome in SPD489-404. The objective of this study was to assess cognitive function over 2 years in study SPD489-404, using the Cambridge Neuropsychological Test Automated Battery (CANTAB). Participants aged 6-17 years received dose-optimised open-label lisdexamfetamine dimesylate (30, 50 or 70 mg/day) for 104 weeks. Cognition was assessed using four CANTAB tasks; Delayed Matching to Sample (DMS), Spatial Working Memory (SWM), Stop Signal Task (SST) and Reaction Time (RTI). Key and additional variables were pre-specified for each CANTAB task; groupwise mean percentage changes in key variables from baseline of > 5% were considered potentially clinically significant. All 314 enrolled participants received lisdexamfetamine dimesylate and were included in the safety population, and 191 (60.8%) completed the study. No potentially clinically significant deteriorations from baseline were observed in any key CANTAB variable over the 2 years of the study. Based on predefined thresholds, potentially clinically significant improvements from baseline were observed at 6 months (DMS median reaction time, mean per cent change, - 6.6%; SWM total between-search errors, - 22.8%; SST stop signal reaction time, -18.9%), and at the last on-treatment assessment (DMS median reaction time, - 6.5%; SWM total between-search errors, - 32.6%; SST stop signal reaction time, - 25.7%). Lisdexamfetamine dimesylate treatment for 2 years was not associated with deterioration of cognitive function in children and adolescents with attention-deficit/hyperactivity disorder. Although improvements in some cognitive measures were observed, lack of a control group makes interpretation of the findings difficult. Further studies of the impact of stimulants on cognition are required. CLINICALTRIALS. NCT01328756.


Impact of placental Plasmodium falciparum malaria on pregnancy and perinatal outcome in sub-Saharan Africa: part III: placental malaria, maternal health, and public health.
Plasmodium falciparum infections of the placenta remain a major medical challenge among pregnant women in sub-Saharan Africa. A number of factors influence the prevalence of placental malaria in pregnant women, including maternal age, gravidity, use of prophylaxis, nutrition, host genetics, and level of anti-parasite immunity, as well as parasite genetics and transmission rates [1]. Maternal anemia has been shown to be one of the major complications of placental malaria in sub-Saharan Africa. The mechanisms by which malaria causes anemia are fairly well understood. The pathophysiology of malaria-associated anemia is multifactorial. The most likely mechanisms include (i) hemolysis or the direct destruction of parasitized red blood cells that occurs both intravascularly and by sequestration in the microcirculation, mainly in the spleen; (ii) specific/nonspecific immune responses, whereby red cell survival is shortened; (iii) nonspecific, defective, red cell production, which depresses erythropoiesis, inhibits reticulocyte release, and prematurely destructs red cells during maturation in the bone marrow; and (iv) hypersplenism associated with a reduction in all three blood cell series, that is, causing not only anemia but also thrombocytopenia and leucopenia [2,3]. The relationship between maternal anemia with obstetric factors, however, is not fully understood, and, thus, evaluating the link between malaria, obstetric disorders, and maternal death has been recommended [4]. There have been efforts to quantify the contribution of malaria to maternal morbidity and mortality with the expectation that this would provide the evidence necessary to improve the effectiveness of advocacy to incorporate malaria prevention strategies in Safe Motherhood Programs [5,6]. The effects of placental malaria on maternal health can better be understood when considered in relation with various maternal parameters, including maternal age, parity, peripheral malaria infection, anemia, and HIV infection.


Effect of dietary ascorbic acid and vitamin E on metabolic changes in rats and guinea pigs exposed to PCB.
The effect of the addition of dietary ascorbic acid and/or vitamin E (all-rac-alpha-tocopheryl acetate) in rats and guinea pigs exposed to PCB (polychlorinated biphenyls) was studied. Rats were fed diets containing one of three levels of vitamin E (30, 500 or 1000 mg/kg diet) with or without PCB (200 mg/kg diet). Guinea pigs were fed diets containing PCB (40 mg/kg diet) with 200 or 1000 mg ascorbic acid/kg diet and/or 70 or 2000 mg vitamin E/kg diet. For rats fed PCB, ascorbic acid in urine was 40-fold higher and in liver, 2-fold higher than for rats fed no PCB, and thiobarbituric acid-reactive substances (TBA-RS, indicators of lipid peroxidation) in liver was 1.5-fold higher. In rats fed PCB, high dietary vitamin E significantly lowered the urinary ascorbic acid and TBA-RS. Liver ascorbic acid was lowered by high dietary vitamin E only in control rats. In guinea pigs, feeding PCB caused severe growth retardation and the liver TBA-RS was 1.8-fold higher than in guinea pigs not fed PCB. Feeding high levels of both ascorbic acid and vitamin E was more effective in reversing the growth depression and in lowering TBA-RS level (due to PCB) than feeding the vitamins separately. Ascorbic acid metabolism in rats was affected by high dietary vitamin E. The possibility of a higher requirement for ascorbic acid and vitamin E in guinea pigs exposed to PCB was indicated. Interaction of ascorbic acid and vitamin E in animals exposed to PCB was suggested.


[Progressive axial exophthalmy--case report].
It is being presented the clinical case of a 67 year old male patient who has been under clinical observation and treatment for 10 years. The diagnosis rests upon clinical methods, imaging techniques (ocular-orbital examination, computerized tomography and magnetic resonance imaging) as well as post-operative histopathology reports. The clinical diagnosis was supported by the progressive decrease in visual acuity, painful, irreducible, non-pulsating axial progressive exophthalmia, associated with disorders of the ocular motility initially at the level of the RE, followed by bilateral involvement. The RE ocular-orbital ultrasound and the MRI examination point out a homogenous retro bulbar formation that caudally compresses and exceeds the optic-nerve. The mass lesion from the right orbit was afterwards partially removed by neurosurgical excision. The general treatment was initiated with steroidal and non-steroidal anti-inflammatory drugs in repetitive cures, under protection of antibiotics and anti-secretory drugs, the response to treatment being unfavorable. In time, the exophthalmia increased progressively, fact that imposed right de-compressive orbitotomy, with the surgical ablation of the lateral orbital wall. About 1 year after the neurosurgical intervention the initial clinical symptoms insidiously reinstalled throughout 3 - 4 years. The ultrasound, completed by the cranium CT and MRI examinations have identified the bilateral presence of myositis. DEBATES: Regardless of all the therapeutical means applied in time, not only the medical treatment (the systemic corticotherapy), but also the surgical one (with palliative effect), the patient's evolution was unfavorable; the axial exophthalmia persists, it is painful and irreducible. The evolution is aggravated also by the fact that both orbits have been affected; the specialty literature mentions cases with frequent unilateral involvement. In the case presented the axial exophthalmia is irreversible; it is determined by an inflammatory pseudotumor of both orbits, that represents a chronic inflammatory, idiopathic disease, with unpredictable clinical evolution. The diagnosis is usually one of exclusion, the complementary imaging examinations being necessary to rule out other pathologies of the orbit.


Effects of core size and PEG coating layer of iron oxide nanoparticles on the distribution and metabolism in mice.
In vivo distribution of polyethylene glycol (PEG)ylated functional nanoparticles is vital for determining their imaging function and therapeutic efficacy in nanomedicine. However, contradictory results have been reported regarding the effect of core size and PEG surface of the nanoparticles on biodistribution. To clarify this ambiguous understanding, using iron oxide nanoparticles (IONPs) as a model system, we investigated the effect of core size and PEG molecule weights on in vivo distribution in mice. Three PEGylated IONPs, including 14 nm IONP@PEG2,000, 14 nm IONP@PEG5,000, and 22 nm IONP@PEG5,000, were prepared with a hydrodynamic size of 26, 34, and 81 nm, respectively. The blood pharmacokinetics and tissue distribution were investigated in detail. The results indicated that the PEG layer, rather than core size, played a dominant role in determining the half-life time of IONPs. Specifically, increased molecular weight of the PEG layer led to a longer half-life time. These PEGylated IONPs were mainly excreted by liver clearance. While the PEG molecular layer constituted the key factor to determine the clearance ratio, core size affected the clearance rate. Complete blood count analysis and histopathology suggested excellent biocompatibility of PEGylated IONPs for future clinical trials.


[The adverse effects of dark sulfonated shale oil following local and systemic administration].
Ichthyol (Ammoniumbituminosulfonat DAB, dark sulfonated shale oil, CAS 8029-68-3) is used for the treatment of inflammatory skin diseases and rheumatic diseases and blunt traumata. The studies conducted were carried out to obtain information on the risk following dermal and oral application of dark sulfonated shale oil. Corresponding to the administration route envisaged for man the drug was applied by topical and systemic administration. Studies were conducted on the acute, subacute and chronic toxicity, local tolerance, and the teratogenic, mutagenic and carcinogenic potential. Dark sulfonated shale oil was found to be well tolerated whether administered for a short period or over a long time. There were no indications for a teratogenic, mutagenic or carcinogenic effect. The therapeutic index (ratio of the toxic dose in animals and the therapeutic human dose) for topical administration of dark sulfonated shale oil is > 7 depending on animal species, frequency of administration, dose-level employed and the toxicological question posed. The results of the toxicological studies coincide with more than 110 years of experience that the drug is well-tolerated by man.


Laparoscopic surgery in a rat model.
The rat is increasingly being used in laparoscopic research. Laparoscopic microsurgical training is critical in order to develop new surgical indications in pediatric patients. This report evaluates laparoscopic splenectomy and nephrectomy in a rat model. A Wistar rat (weight between 250 and 400 g) was placed in the supine position. Inhaled 3% halothane anesthesia was administered. A Veress needle is inserted in the right-upper abdomen. After establishing a pneumoperitoneum of 3-4 mm Hg, a 2- or 5-mm trocar was placed, according to the procedure. A 2-mm 0-degree endoscope was used. Two additional 2- or 5-mm trocars were then placed. Laparoscopic splenectomy involved two-handed dissection, intracorporeal ligation, and the division of gastrosplenic attachments and hilar and short gastric vessels. Laparoscopic nephrectomy was done by intracorporeal ligation and division of the renal vessels and the ureter after mobilization of the kidney. Laparoscopic splenectomy was performed in 8 rats; laparoscopic nephrectomy was done in 4 rats. Operative time was 25-40 minutes for splenectomy and from 30 to 65 for nephrectomy. Postoperatively, 4 rats died from hemorrhage. No wound infections occurred at the port sites. Laparoscopic splenectomy and nephrectomy in an experimental rat model is technically feasible and may provide an excellent training model for pediatric minimally invasive surgery.


Towards a highly efficient theoretical treatment of Jahn-Teller effects in molecular spectra: the 1 2A and 2 2A electronic states of the ethoxy radical.
Nonadiabatic effects in the two lowest electronic states of the ethoxy radical, the 1 (2)A and 2 (2)A states, are considered, using multireference configuration interaction (MRCI) wave functions comprised of over 15x10(6) configuration state functions. The lowest point on the seam of conical intersection is located. Using this point as the origin, a quasidiabatic Hamiltonian suitable for use in a multimode vibronic coupling treatment of the coupled 1 (2)A and 2 (2)A electronic states is determined. The Hamiltonian includes all contributions from all internal coordinates through second order in displacements from the origin and is comprised of over 500 parameters. By using the average energy gradient, the energy difference gradients, and the derivative couplings, all of which are obtained at little additional cost once the requisite eigenstates are known, the second order Hamiltonian is determined from MRCI calculations at only 35 nuclear configurations. This is essentially the same number of points required to obtain the frequencies for the ground state equilibrium structure using centered differences of gradients. The diabatic Hamiltonian provides a good description of the seam space, the (N(int)-2)-dimensional space of conical intersection points, continuously connected to the minimum energy crossing point, enabling, for the first time, an analysis of the changes in the branching plane induced by seam curvature in the full seam space. Comparing the diabatic representation and MRCI results we find a good agreement for the ground state equilibrium structure, R(eq)(1 (2)A), as well as the ground state energy and vertical excitation energy. In good agreement with the available experimental data are the ground state equilibrium structure and the excitation energy to the A (2)A state, predicted here to involve a cone state level. Agreement between the harmonic frequencies at R(eq)(1 (2)A) computed from the MRCI wave function and from the diabatic Hamiltonian is excellent for all but the three lowest energy normal modes where significant deviations are observed indicating the need for selected cubic and/or quartic terms. For the low-lying vibrational levels, the diabatic representation can be used to partition the normal modes into two groups, those that involve inter(diabatic) state coupling and those that are spectators as far as nonadiabatic effects are concerned. The spin-orbit coupling interaction is determined using the Breit-Pauli approximation and its incorporation into the diabatic Hamiltonian is discussed.


Exploring the role of antithrombin replacement for the treatment of preeclampsia: a prospective randomized evaluation of the safety and efficacy of recombinant antithrombin in very preterm preeclampsia (PRESERVE-1).
Antithrombin (AT) replacement has been described in patients with hereditary AT deficiency undergoing delivery; however, the kinetics of AT replacement in preeclampsia is not adequately understood. Therefore, the Prospective Randomized Evaluation of the Safety and Efficacy of Recombinant Antithrombin in Very Preterm Preeclampsia (PRESERVE-1) study has been proposed. Sixty women aged≥18 years at 24 0/7-28 0/7 weeks' gestation and with hypertension and proteinuria will be enrolled and randomly assigned to receive recombinant human AT or placebo until fetal and/or maternal indications cause cessation of expectant management or until 34 0/7 weeks' gestation. The primary endpoint is the increase in gestational age from randomization to delivery. Safety assessments and laboratory assays will also be performed. PRESERVE-1 study enrollment will begin during the second half of 2013. The PRESERVE-1 study will provide further insight into the pharmacokinetic activity and safety of AT therapy in preeclampsia.


Multiple serotonergic mechanisms contributing to sensitization in aplysia: evidence of diverse serotonin receptor subtypes.
The neurotransmitter serotonin (5-HT) plays an important role in memory encoding in Aplysia. Early evidence showed that during sensitization, 5-HT activates a cyclic AMP-protein kinase A (cAMP-PKA)-dependent pathway within specific sensory neurons (SNs), which increases their excitability and facilitates synaptic transmission onto their follower motor neurons (MNs). However, recent data suggest that serotonergic modulation during sensitization is more complex and diverse. The neuronal circuits mediating defensive reflexes contain a number of interneurons that respond to 5-HT in ways opposite to those of the SNs, showing a decrease in excitability and/or synaptic depression. Moreover, in addition to acting through a cAMP-PKA pathway within SNs, 5-HT is also capable of activating a variety of other protein kinases such as protein kinase C, extracellular signal-regulated kinases, and tyrosine kinases. This diversity of 5-HT responses during sensitization suggests the presence of multiple 5-HT receptor subtypes within the Aplysia central nervous system. Four 5-HT receptors have been cloned and characterized to date. Although several others probably remain to be characterized in molecular terms, especially the Gs-coupled 5-HT receptor capable of activating cAMP-PKA pathways, the multiplicity of serotonergic mechanisms recruited into action during learning in Aplysia can now be addressed from a molecular point of view.


[Allogenic stem cell transplantation from genotypically HLA-identical siblings for 30 patients with myelodysplastic syndromes].
To explore the indication and optimum time for treating myelodysplastic syndrome (MDS) by allogeneic hematopoietic stem cell transplantation (allo-HSCT) with HLA identical sibling grafts. From June 1997 to Sep. 2004, a total of 30 patients with MDS were treated with allo-HSCT from HLA-identical sibling donors in our institute. On HSCT, 4 patients had refractory anemia (RA) , 2 RA with ringed sideroblasts (RARS) , 7 RA with excess blasts(RAEB) , 14 RAEB in transformation (RAEB-t) , 3 already progressed to secondary AML. For IPSS system, 6 patients were in intermediate- I risk group, 11 in intermediate- li risk group, and 13 in high risk group. The modified BU/CY conditioning regimen was used. Four patients received bone marrow transplantation (BMT), 8 received peripheral blood stem cell transplantation (PBSCT) , and 18 received BMT + PBSCT. The 3-year expected overall survival (OS) was 63.61%, 3-year expected disease-free survival ( DFS) 61.41%, and relapse rate 5.26%; OS for RA/ RAS, RAEB and RAEB-t/AML subgroup was 83.33%, 34.29% and 66.67% , respectively, and all had no statistic difference among them. OS for IPSS-intermediate and high risk subgroup was 64.7% , and 69.0% respectively, also had no statistic difference. 3-year expected OS in no aGVHD,grade I - II aGVHD and grade III - IV aGVHD group was 57.75% , 100% and 0% , respectively (P = 0.009). Pre-HSCT chemotherapy, disease subtype and cGVHD all had no correlation with LFS or OS (P > 0.05). For young MDS patients having HLA-identical sibling donors, HSCT should be the first line therapy and performed as soon as possible.


Ship source level estimation and uncertainty quantification in shallow water via Bayesian marginalization.
This paper applies a non-linear Bayesian marginalization approach to ship spectral source level estimation in shallow water with unknown seabed properties and uncertain source depth. The algorithm integrates the posterior probability density over seabed models sampled via trans-dimensional Bayesian matched-field inversion and over depths/ranges of multiple point sources (representing different noise-generating components of a large ship) via Metropolis-Hastings sampling. Source levels and uncertainty are derived from marginal distributions for source strength. The approach is applied to radiated noise due to a container ship recorded on a bottom-moored horizontal array in shallow water. The average uncertainty is 3.8 dB/Hz for tonal frequencies.


A synthesized GFP analogue emits via a hydrogen-bonding system.
Two GFP analogues o-HBDI and o-HBMO have been designed and synthesized. The former shows no fluorescence while the latter shows strong fluorescence due to the formation of an intramolecular hydrogen-bond. At low temperatures, both molecules have strong fluorescence.


Nasal Septal Abscess Developed After Radiofrequency Cauterization for Epistaxis.
Nasal septal abscesses are commonly caused by nasal trauma, infection, and nasal surgery. As delayed treatment of nasal septal abscesses can cause various complications, including saddle nose, a prompt diagnosis with the relevant intervention is important. However, the diagnosis of nasal septal abscesses is difficult when nasal pain occurs after cauterization because there have been no formal reports of nasal septal abscesses owing to cauterization for epistaxis. Here, the authors report the first case of a nasal septal abscess that developed after cauterization. A 48-year-old woman underwent radiofrequency cauterization as a treatment for epistaxis. She developed nasal pain 10 days following cauterization, and a computed tomography scan revealed a nasal septal abscess. Careful observation should be undertaken in patients who undergo cauterization of the nasal septum, especially in patients complaining of nasal pain.


Identification of titanium dioxide nanoparticles in food products: induce intracellular oxidative stress mediated by TNF and CYP1A genes in human lung fibroblast cells.
Food grade TiO2 (E171) is a synthetic additive, and widely used as a coloring agent in many foods, pharmaceutical and personal care products. A few reports have highlighted that insoluble particulates (less than 200nm) of food grade TiO2 are found in many foods and confectionary products. However, information regarding the physico-chemical properties (i.e., size and shape)-based food grade TiO2 nanotoxicity related human health issues are limited. The main goal of this study is to examine the presence of nano-sized particulates and its structural characteristics of food grade- TiO2 materials and to assess the acute cellular uptake and metabolic stress induced by these particulates in human lung fibroblast (WI-38) cells. The results of transmission electron microscopy, energy dispersive X-ray spectroscopy, and X-ray diffraction studies indicated that about food grade TiO2 sample contains spherical shaped particulate forms in the nano-scale range, <100nm. The intracellular oxidative stress in human lung fibroblast cells (WI-38) was assessed through studies investigating the cellular uptake of the particles, changes in nuclear and cytoplasmic morphology, intracellular ROS, mitochondrial trans-membrane potential, the cell cycle and the expression of genes linked to metabolic stress markers. Altogether our data clearly indicate that primary metabolic stress indicators such as changes in the intracellular ROS, the dose-dependent loss of the mitochondrial membrane potential, alterations in cell cycle progression (G2/M>S>G0/G1) and changes in the TNF and CYP1A gene expression pattern are linked to cellular stress. Thus, food grade TiO2 as nano-scaled contaminants could not only be potential human health risk factors, suggesting that safety considerations with special respect to a few crucial factors such as size, and shape should be considered and regulated by food regulators.


Simultaneous measurement of uptake and elimination of cadmium by caddisfly (Trichoptera: Hydropsychidae) larvae using stable isotope tracers.
The use of stable isotopes coupled with inductively coupled plasma-mass spectrometry provides a unique opportunity to investigate pathways of metals in aquatic invertebrates. In this study, the simultaneous uptake and elimination of cadmium (Cd) in stream-dwelling caddisfly (Trichoptera: Hydropsychidae) larvae was measured in laboratory experiments with stable isotopes of Cd. In one experiment, animals were exposed to low levels (0.5 microg/L) of 114Cd in filtered river water and uptake was measured for 9 d, followed by 9 d during which the larvae were exposed to either 114Cd, 113Cd, or filtered river water. In a second experiment, the initial exposure concentration of 114Cd varied from 0.1 to 1.6 microg/L for 5 d, followed by 5 d during which the larvae were exposed to the same respective levels of 113Cd. The purpose of the two experiments was to test whether or not elimination of 114Cd from Hydropsyche larvae was the same in the presence or absence of Cd in the water and also whether or not elimination rates were dependent on exposure concentration. The results indicated that elimination of Cd by Hydropsyche larvae (elimination rate constant approximately 0.21-0.24/d) in the presence of Cd in the water was the same as the depuration when no metal was present. Furthermore, the elimination rate was independent of exposure concentrations, ranging between 0.1 and 1.6 microg/L. Thus, the stable isotope tracer technique enabled the simultaneous measurement of uptake and elimination of Cd in hydropsychid larvae and may have the potential to facilitate the labeling of various compartments (e.g., water, sediment, and food) in the aquatic environment.


Lack of correlation between length variation in the DNA polymerase gamma gene CAG repeat and lactic acidosis or neuropathy during antiretroviral treatment.
Antiretroviral therapy, although successful in reducing HIV load and accordingly decreasing the incidence of HIV infection-related symptoms, has its drawbacks in the form of severe side effects. Recognized drug-related side effects are, for example, nausea, fatigue, lactic acidosis, neuropathy, lipodystrophy, and myopathy. Because not all patients experience these side effects, genetic factors could be involved. It is believed that the main toxicity of nucleoside analog drugs is due to a decrease in mitochondrial function, possibly by inhibition of mitochondrial DNA (mtDNA) replication. mtDNA is replicated by a multienzyme complex, the main component of which is the nuclear-encoded DNA polymerase gamma. Presently, the only known variation in the DNA polymerase gamma gene is variation in the number of CAG repeats in the second exon. To investigate whether CAG repeat expansion or mutations in the DNA polymerase gamma (POLG) gene could predispose to peripheral neuropathy or lactic acidosis, we have sequenced part of the second exon of the DNA polymerase gamma gene, containing the CAG repeat, of 59 drug-treated HIV-infected patients, 11 of whom experienced drug-induced neuropathy, and 3 of whom died from lactic acidosis. No correlation was found between numbers of CAG repeats and any of the symptoms. The coding regions of the POLG gene from the three lactic acidosis patients were then completely sequenced, but no mutations were found. In addition, no variation was detected in exons 3, 8, and 19 of seven neuropathy patients and three control subjects without symptoms. These exons were the only sites of amino acid changes between human and chimpanzee POLG genes, and were chosen as targets of tolerated variation.


Validity of linear response theory for time-dependent fluorescence in Staphylococcus nuclease.
We report a theoretical study on the relaxation dynamics of Staphylococcus nuclease following photon excitation. Molecular dynamics simulations were implemented for both ground state and excited state surface of Trp140. Over the course of 40 ns ground state simulation, a structural transition was observed from one isomeric protein configuration to another. No obvious isomerization process was exhibited in the excited state simulations. Using linear response theory and direct nonequilibrium approach, time-dependent Stokes shift, as well as its time-infinity value, was evaluated for both ground state isomers. Comparison between these methods exhibits severe disagreement. The nonequilibrium simulations show the similar relaxation dynamics as the excited state linear response approach, whereas they severely differ from the ground state linear response evaluation. Further examination reveals that the isomerization process breaks the characteristics of the energy gap histogram from Gaussian statistics. We also made the comparison between the experimental and simulation results. The significant inertial decay, in theory, is absent in the experiment. The ground state linear response evaluation provides a better agreement with the experimental results among all three approaches.


The roles of annexins in vascular endothelium dysfunction accompanying diabetes mellitus type 2.
Impairment in cellular transport, distribution and storage of cholesterol accompanies insulin resistance and diabetes mellitus type 2 as well as other diseases such as obesity, atherosclerosis, and non-alcoholic fatty liver disease. Diabetes mellitus type 2 is a metabolic disorder that is characterized by hyperglycemia in the context of insulin resistance and relative lack of insulin. Type 2 diabetes makes up about 90% of cases of diabetes. Several therapeutic strategies are today being considered to target diabetes mellitus type 2, and the accompanying endothelial dysfunction, but none as yet has proved satisfactory. Accumulating data suggest that annexins, as cholesterol binding proteins that participate in intracellular transport and storage of cholesterol and in the organization of plasma membrane, may participate in development and sustenance of diabetes mellitus type 2 and may serve as predictive markers of this disease.


C-terminal cyclization of an SDF-1 small peptide analogue dramatically increases receptor affinity and activation of the CXCR4 receptor.
In an effort to improve the activities and bioavailabilities of stromal cell-derived factor-1 (SDF-1, CXCL12) sdf-(1-67)-OH (1), we have prepared a linear peptide analogue [sdf-(1-31)-NH(2) (2)] and two lactam analogues [cyclo(Lys(20)-Glu(24))-sdf-(1-31)-NH(2) (3) and cyclo(Glu(24)-Lys(28))-sdf-(1-31)-NH(2) (4)], consisting of the N-terminal region (amino acids 1-14) joined by a four-glycine linker to the C-terminal region (amino acids 56-67) of 1. Analogues 2 and 4 had eight residues of alpha-helix, as estimated from its circular dichroism (CD) spectra, in contrast to 10 residues in analogue 3. Cyclization of analogue 2 at residues 20 and 24 to give analogue 3 resulted in only a slight change to the theta;(222)/theta;(209) ratio (0.81 to 0.86, where 1.09 is considered a perfect alpha-helix), although an increase in the alpha-helix length of analogue 3 was observed. In contrast, cyclization between residues 24 and 28 by lactamization to give analogue 4 only slightly affected the helical content but clearly resulted in a more classical alpha-helical structure (theta;(222)/theta;(209) = 0.98). Cyclization of the linear analogue 2 enhanced the SDF-1 receptor CXCR4 binding approximately 114-fold, where the IC(50) values derived from (125)I-SDF-1 competitive binding assays with CEM cells were found to be 39.5 +/- 5.9 nM, 28.9 +/- 6.3 microM, 225.8 +/- 11.8 nM, and 254.1 +/- 5.4 nM for analogues 1-4, respectively. Intracellular calcium mobilization ([Ca(2+)](i)) induced after interaction with CXCR4, as measured by EC(50), was significantly reduced in analogue 4 compared to 3, and approached the EC(50) of native SDF-1, indicating a correlation between the degree of alpha-helix and biological activity. Therefore, the biological activity of small peptide SDF-1 analogues is highly dependent on the conformation of its C-terminal region.


Scanning gate imaging of two coupled quantum dots in single-walled carbon nanotubes.
Two coupled single wall carbon nanotube quantum dots in a multiple quantum dot system were characterized by using a low temperature scanning gate microscopy (SGM) technique, at a temperature of 170 mK. The locations of single wall carbon nanotube quantum dots were identified by taking the conductance images of a single wall carbon nanotube contacted by two metallic electrodes. The single electron transport through single wall carbon nanotube multiple quantum dots has been observed by varying either the position or voltage bias of a conductive atomic force microscopy tip. Clear hexagonal patterns were observed in the region of the conductance images where only two sets of overlapping conductance rings are visible. The values of coupling capacitance over the total capacitance of the two dots, C(m)/C(1(2)) have been extracted to be 0.21 ∼ 0.27 and 0.23 ∼ 0.28, respectively. In addition, the interdot coupling (conductance peak splitting) has also been confirmed in both conductance image measurement and current-voltage curves. The results show that a SGM technique enables spectroscopic investigation of coupled quantum dots even in the presence of unexpected multiple quantum dots.


Simple reaction times in subjects with steeply sloping hearing loss: is there an alteration at the edge of the loss?
The aim of the present study was to investigate the simple reaction time (RT) performance of patients with steeply sloping sensorineural hearing loss. This kind of hearing loss has the particularity of inducing a reorganization of the primary auditory cortex (Dietrich et al, 2001), the edge frequency of the loss being over-represented. It is assumed that a consequence of this plasticity could be reflected in an improvement in frequency discrimination performances around the edge of the loss (McDermott et al, 1998). In the present study we used pure tones equalized in loudness to investigate whether RT might be altered at the cut-off frequency (Fc) of the loss, or at the frequency that presented the best discrimination-limen-for-frequency (bDLF). The effect of auditory rehabilitation on RT was also studied. A great variability in RT was noted in all our subjects. Our results demonstrated that RT was unmodified at Fc or bDLF, be it before or after hearing aid fitting. However, an improvement in the correlation between RT and frequency discrimination performance was observed at three months post-rehabilitation.


Discrepancies in Ratings of Behavioral Healthcare Interventions Among Evidence-Based Program Resources Websites.
Decision makers in the behavioral health disciplines could benefit from tools to assist them in identifying and implementing evidence-based interventions. One tool is an evidence-based program resources website (EBPR). Prior studies documented that when multiple EBPRs rate an intervention, they may disagree. Prior research concerning the reason for such conflicts is sparse. The present study examines how EBPRs rate interventions and the sources of disagreement between EBPRs when rating the same intervention. This study hypothesizes that EBPRs may disagree about intervention ratings because they either use different rating paradigms or they use different studies as evidence of intervention effectiveness (or both). This study identified 15 EBPRs for inclusion. One author (M.J.L.E.) coded the EBPRs for which "tiers of evidence" each EBPR used to classify behavioral health interventions and which criteria they used when rating interventions. The author then computed one Jaccard index of similarity for the criteria shared between each pair of EBPRs that co-rated interventions, and one for the studies used by EBPR rating pairs when rating the same program. The authors used a combination of chi-square, correlation, and binary logistic regression analyses to analyze the data. There was a statistically significant negative correlation between the number of Cochrane Risk of Bias criteria shared between 2 EBPRs and the likelihood of those 2 EBPRs agreeing on an intervention rating (r = -.12, P ≤ .01). There was no relationship between the number of studies evaluated by 2 EBPRs and the likelihood of those EBPRs agreeing on an intervention rating. The major reason for disagreements between EBPRs when rating the same intervention in this study was due to differences in the rating criteria used by the EBPRs. The studies used by the EBPRs to rate programs does not appear to have an impact.


Antithrombin-III prevents the lethal effects of Escherichia coli infusion in baboons.
Infusion of Escherichia coli (LD100) was followed by coagulopathic and cell injury responses, cardiovascular collapse, and death in 18 to 32 hr in four out of four baboons. Infusion of AT-III in sufficient amounts to achieve AT-III levels of more than 4 units/ml of plasma before and during the infusion of E. coli reduced the intensity of the coagulopathic and cell injury response and prevented vascular collapse and death in four out of four baboons. Failure to achieve AT-III levels of more than six units/ml at T +60 min during the infusion of E. coli resulted in failure to prevent its lethal effects in three out of three baboons even though levels as high as 10 units/ml were achieved later in the course of the experiment. These studies suggest that thrombin and/or its products can contribute to the inflammatory response to E. coli and that AT-III is of potential value as a prophylactic but not as a therapeutic agent in the treatment of patients at high risk of developing gram negative sepsis.


Magnesium Sulfate as an Alternative In Vitro Anticoagulant for the Measurement of Platelet Parameters?
There are conflicting reports on the reliable measurement of platelet count and mean platelet volume (MPV) using EDTA or citrate. The anticoagulant properties of magnesium sulfate (MgSO4) are known from the literature. The aim of this study was to evaluate MgSO4 as an in vitro anticoagulant for platelet count, MPV, platelet distribution width, and platelet activation. Whole blood from volunteers was anticoagulated by EDTA, citrate, or MgSO4 Platelets were counted by the XE 5000 (Sysmex, Norderstedt, Germany) impedance and fluorescence optical technique. The mean impedance platelet counts were 227.7, 197.0, and 201.1 × 10(9)/L in EDTA-, citrate-, or MgSO4-anticoagulated blood, respectively. The counts were 4.7% higher (EDTA) after 3 hours of storage but 4% lower in citrate-anticoagulated blood. The counts in magnesium samples remained stable. The MPV was 10.4 fL (EDTA), 9.5 fL (citrate), and 9.3 fL (MgSO4). EDTA samples showed cell swelling within the first 3 hours. This was lower in citrate and only marginal in magnesium samples. High activation of platelets was observed only in EDTA samples. Magnesium anticoagulation might be advantageous for more reliable MPV measurements. Although platelet count is underestimated when the impedance method is used, the platelet count reveals similar results when measured by the fluorescent optical method.


Pharmacokinetics of calcium antagonists under development.
Calcium antagonist drugs under clinical development are of the Type I (verapamil, diltiazem-like) and Type II (nifedipine-like) classes. Tiapamil, the only Type I drug currently available, is a high clearance, widely distributed drug which undergoes extensive presystemic elimination. Pharmacokinetically it is quite similar to verapamil; however, it does have increased biliary excretion and decreased binding to plasma proteins. Eight Type II (dihydropyridine) drugs are reviewed. Seven of these drugs (felodipine, isradipine, nicardipine, nilvadipine, nimodipine, nisoldipine and nitrendipine) are pharmacokinetically similar to nifedipine, with high clearance, extensive distribution, and significant presystemic elimination. Amlodipine has lower clearance, even greater peripheral distribution, and greatly decreased presystemic elimination. Three of the 8 dihydropyridines have been reported to have plasma protein binding greater than 90%. Unlike nifedipine, each dihydropyridine drug under development has an asymmetric centre; therefore each in fact is a racemic mixture. Human pharmacokinetic and pharmacodynamic data have not been reported for any of the racemates. Each of the drugs has been studied in patients with hepatic and renal disease. Predictably, patients with severe hepatic disease have decreased presystemic clearance and, in some cases decreased clearance after intravenous administration of the dihydropyridines, although renal failure has little influence on their pharmacokinetics. Unfortunately, disease-drug interaction studies of this class of drugs do not generally report plasma protein binding. The effect of age on the disposition of 2 of the dihydropyridines has been reported; however, only for nicardipine can a conclusion be drawn, namely that volume of distribution may increase with age and clearance may remain unchanged. A variety of potential drug-drug interactions have been evaluated, most commonly the effect of these drugs on cardiac glycoside disposition and effect, and the effect of cimetidine on the disposition of dihydropyridines. Tiapamil, like verapamil, impairs digoxin clearance significantly. Among the dihydropyridines, although minor pharmacokinetic effects have in some cases been reported, the magnitude of the interactions suggest they have limited clinical importance. From drugs currently under development, it is clear that a large number of calcium antagonists will soon be introduced into clinical use. Only 1 of the newer drugs, amlodipine, has significant pharmacokinetic differences from the agents currently in use, although possible pharmacodynamic differences among the drugs have been suggested.(ABSTRACT TRUNCATED AT 400 WORDS)


The nucleotide sequence of citrus leaf rugose ilarvirus RNA-2.
The nucleotide sequence of citrus leaf rugose ilarvirus (CiLRV) RNA-2 consists of 2990 nucleotides and contains one open reading frame (ORF) which encodes a deduced translation product of 832 amino acids with a calculated M(r) of 95,501 (95K). The 5' terminus of the RNA has a m7Gppp cap. Both the nucleotide sequence of CiLRV RNA-2 and its translated polypeptide share homologies with the nucleotide sequence and translated polypeptide, respectively, of RNA-2 of alfalfa mosaic virus (AlMV). The homology occurs in the central region of both the nucleic acid sequence and the polypeptide. Homologies between either CiLRV or AlMV and other Bromoviridae (cucumber mosaic virus--CMV, brome mosaic virus--BMV and cowpea chlorotic mottle virus--CCMV) were far less. Alignment of the 104 amino acid region (polymerase signature) of the 95K protein against 10 other 'alpha-like' plant viral polymerase signatures showed that CiLRV and AlMV are more closely related to each other than to CMV, BMV or CCMV. This is the first report of the full-length sequence of RNA-2 of an ilarvirus.


Can CH⋅⋅⋅π interactions be used to design single-chain magnets?
This theoretical study suggests that CH⋅⋅⋅π stacking interactions between monomeric units can be used to design novel single-chain magnets (SCMs), as the sign of coupling is predictable and such chains inherently yield negative axial anisotropy, a condition often difficult to achieve in conventional SCMs.


Geographical distribution of certain cancers in South Africa, 1968-1972.
The geographical distributions of six major causes of cancer mortality in South Africa have been analysed for three population groups: White, Coloured and Asian. Because of the lack of comparable data, the majority Black population is not included. Geographical patterns are compared and discussed in the light of findings from other countries. In several cases the potential for aetiological follow-up is demonstrated.


Alkaloids from Microcos paniculata with cytotoxic and nicotinic receptor antagonistic activities.
Microcos paniculata is a large shrub or small tree that grows in several countries in South and Southeast Asia. In the present study, three new piperidine alkaloids, microgrewiapines A-C (1-3), as well as three known compounds, inclusive of microcosamine A (4), 7'-(3',4'-dihydroxyphenyl)-N-[4-methoxyphenyl)ethyl]propenamide (5), and liriodenine (6), were isolated from cytotoxic fractions of the separate chloroform-soluble extracts of the stem bark, branches, and leaves of M. paniculata. Compounds 1-6 and 1a (microgrewiapine A 3-acetate) showed a range of cytotoxicity values against the HT-29 human colon cancer cell line. When evaluated for their effects on human α3β4 or α4β2 nicotinic acetylcholine receptors (nAChRs), several of these compounds were shown to be active as nAChR antagonists. As a result of this study, microgrewiapine A (1) was found to be a selective cytotoxic agent for colon cancer cells over normal colon cells and to exhibit nicotinic receptor antagonistic activity for both the hα3β4 and hα4β2 receptor subtypes.


Territorial defense, territory size, and population regulation.
The carrying capacity of an environment is determined partly by how individuals compete over the available resources. To territorial animals, space is an important resource, leading to conflict over its use. We build a model where the carrying capacity for an organism in a given environment results from the evolution of territorial defense effort and the consequent space use. The same evolutionary process can yield two completely different modes of population regulation. Density dependence arises through expanding and shrinking territories if fecundity is low, breeding success increases gradually with territory size, and/or defense is cheap. By contrast, when fecundity is high, breeding success sharply saturates with territory size, and/or defense is costly, we predict fixed territory sizes and regulation by floaters. These "surplus" individuals form a buffer against population fluctuations. Yet floaters can also harm breeder performance, and by comparing population growth of a territorial population to a nonterritorial (and individually suboptimal) alternative, we can quantify the harmful effect of evolutionary conflict on population performance. Territoriality has often been found to increase population stability, but this may come at a cost of reduced equilibrium densities.


Effects of Surgical Assistant's Level of Resident Training on Surgical Treatment of Intermittent Exotropia: Operation Time and Surgical Outcomes.
To evaluate the effects of the surgical assistant's level of resident training on operation time and surgical outcome in the surgical treatment of intermittent exotropia. This study included 456 patients with intermittent exotropia who underwent lateral rectus recession and medial rectus resection and were followed up for 24 months after surgery. The patients were divided into two groups according to the surgical assistant's level of resident training: group F (surgery assisted by a first-year resident [n = 198]) and group S (surgery assisted by a second-, third-, or fourth-year resident [n = 258]). The operation time and surgical outcomes (postoperative exodeviation and the number of patients who underwent a second operation) were compared between the two groups. The average operation times in groups F and S were 36.54 ± 7.4 and 37.34 ± 9.94 minutes, respectively (p = 0.33). Immediate postoperative exodeviation was higher in group F (0.79 ± 3.82 prism diopters) than in group S (0.38 ± 3.75 prism diopters). However, repeated-measures analysis of variance revealed no significant difference in exodeviation between the two groups during the 24-month follow-up period (p = 0.45). A second operation was performed in 29.3% (58 / 198) of the patients in group F, and in 32.2% (83 / 258) of those in group S (p = 0.51). No significant difference in operation time was observed when we compared the effects of the level of resident training in the surgical treatment of intermittent exotropia. Although the immediate postoperative exodeviation was higher in patients who had undergone surgery assisted by a first-year resident, the surgical outcome during the 24-month follow-up was not significantly different.


Intracellular metabolic pathway distribution in diatoms and tools for genome-enabled experimental diatom research.
Diatoms are important primary producers in the oceans and can also dominate other aquatic habitats. One reason for the success of this phylogenetically relatively young group of unicellular organisms could be the impressive redundancy and diversity of metabolic isoenzymes in diatoms. This redundancy is a result of the evolutionary origin of diatom plastids by a eukaryote-eukaryote endosymbiosis, a process that implies temporary redundancy of functionally complete eukaryotic genomes. During the establishment of the plastids, this redundancy was partially reduced via gene losses, and was partially retained via gene transfer to the nucleus of the respective host cell. These gene transfers required re-assignment of intracellular targeting signals, a process that simultaneously altered the intracellular distribution of metabolic enzymes compared with the ancestral cells. Genome annotation, the correct assignment of the gene products and the prediction of putative function, strongly depends on the correct prediction of the intracellular targeting of a gene product. Here again diatoms are very peculiar, because the targeting systems for organelle import are partially different to those in land plants. In this review, we describe methods of predicting intracellular enzyme locations, highlight findings of metabolic peculiarities in diatoms and present genome-enabled approaches to study their metabolism.This article is part of the themed issue 'The peculiar carbon metabolism in diatoms'.


Youth's Adjustment to Cancer: Examination of Patterns of Adjustment and the Role of Peer Relations.
Examine unique forms of peer relations (i.e., peer group vs. friendships) in relation to patterns of youth's resilience and challenge-related growth in the context of cancer. In all, 279 youth (cancer, n = 156; control, n = 123) completed measures of posttraumatic stress, depression, anxiety, posttraumatic growth (PTG), and perceived positive changes. Youth also reported on their peer relations. Latent profile analysis (LPA) was used to examine patterns of youth's adjustment. Peer relations were examined as predictors of youth's adjustment. LPA revealed three profiles (42.1% resilient high growth, 21.4% resilient low growth, and 36.5% mild distress with growth). Youth's peer relations, demographic factors, and disease-related factors predicted assignment to profiles. Differences in adjustment emerged depending on youth's connection with their peers versus their friends. Peer relations serve an important role in youth's adjustment to stressful life events. Assessment of peer and friend support may provide a more nuanced understanding of adjustment processes.


[The protective role of peroxisome proliferator activated receptors-α and pathomechanism in D-galactosamine/lipopolysaccharide- induced acute liver failure in rats].
To determine the role and mechanism of peroxisome proliferator activated receptors (PPAR) α in a mouse model of D-galactosamine/lipopolysaccharide (D-GalN/LPS)-induced acute liver failure (ALF). Firstly, C57BL/6 mice were randomly divided into control group (n = 8), ALF 2h group (n = 8), ALF 4h group (n = 8), ALF 6h group (n = 8). Secondly C57BL/6 mice were randomly divided into control group (n = 8), ALF group (n = 8), WY14643 group (n = 8). To induce ALF, the mice were injected intraperitoneally with D-GalN (700 mg/kg) and LPS (10 µg/kg). WY14643 (6 mg/kg), the selective agonist of PPAR α, was administered via tail vein two hours prior to D-GalN/LPS exposure. Two, four, and six hours after D-GalN/LPS treatment in the first study, mice were anesthetized and blood was collected, 6h after D-GalN/LPS treatment in the second study, blood was collected. The liver tissue was harvested for histology and mRNA extraction. Serum levels of ALT and AST were measured to evaluate the hepatic damage. Inflammatory cytokines (TNFα, IL-1β, IL-6) and chemokines (CXCL-1, CXCL-10) were detected by real-time quantitative PCR. Differential protein expression of p-NF-κBp65, p-JNK, p-ERK, p-p38 in inflammatory pathways was detected by Western blotting. Significance of inter-group differences was assessed by one-way ANOVA, and pairwise comparison was performed by the least significant difference test. The gene and protein expression of PPAR α were gradually reduced during the development of ALF. Compared with the model group, the liver architecture was better preserved almost with normal morphology in WY14643-treated mice. Serum ALT and AST levels in WY14643-treated group were significantly lower [ALT: (555 ± 62)U/L vs (2 898 ± 822) U/L, P < 0.05; AST: (791 ± 58) U/L vs (3 013 ± 997) U/L, P < 0.05]. The expression of proinflammatory cytokines and chemokines was significantly suppressed during the activation of PPAR α. In the second study, the levels of gene expression of proinflammatory cytokines and chemokines were detected in control group, ALF group and WY14643 group respectively as followings: TNFα (0.161 ± 0.085, 7.996 ± 1.068, 3.346 ± 0.94, P < 0.05), IL-1β (0.041 ± 0.002, 3.657 ± 0.904, 0.176 ± 0.089, P < 0.01), IL-6 (0.018 ± 0.008, 1.762 ± 0.589, 0.163 ± 0.0487, P < 0.05), CXCL-1 (0.063 ± 0.008, 7.881 ± 0.966, 2.737 ± 0.864, P < 0.01), CXCL-10 (0.054 ± 0.005, 5.671 ± 0.948, 2.578 ± 0.804, P < 0.05). Our findings first demonstrate that PPARα protects liver from injury in an ALF mouse model by suppressing inflammatory response, indicating PPARα as a potential new therapeutic target for ALF.


The relative biological effectiveness of the modulated proton beam at TRIUMF.
The purpose of this study was to determine the relative biological effectiveness (RBE) of proton beam and to study its dependency on fraction number. The relative biological effectiveness (RBE) of fractionated protons compared with 60Co gamma-rays was investigated for the acute mouse skin reaction. The 80 MeV protons generated at the TRIUMF cyclotron were spread out from 7 to 25 mm to irradiate entire legs. One, two, four, and eight fractions were tested. RBE values ranged from 1.15 to 1.24 for the acute skin reaction. Fraction dose dependence of RBEs was not observed.


Virtual screening and bioassay study of novel inhibitors for dengue virus mRNA cap (nucleoside-2'O)-methyltransferase.
We report high-throughput structure-based virtual screening of putative Flavivirus 2'-O-methyltransferase inhibitors together with results from subsequent bioassay tests of selected compounds. Potential inhibitors for the S-adenosylmethionine binding site were explored using 2D similarity searching, pharmacophore filtering and docking. The inhibitory activities of 15 top-ranking compounds from the docking calculations were tested on a recombinant methyltransferase with the RNA substrate (7Me)GpppAC(5). Local and global docking simulations were combined to estimate the ligand selectivity for the target site. The results of the combined computational and experimental screening identified a novel inhibitor, with a previously unknown scaffold, that has an IC(50) value of 60 microM.


Oxygenated Hemoglobin Predicts Outcome in Patients with Chronic Lung Allograft Dysfunction.
Long-term outcome of lung transplantation (LTx) recipients is limited by chronic lung allograft dysfunction (CLAD). In this setting of new onset respiratory failure, the amount of oxygenated hemoglobin (OxyHem; hemoglobin (Hb) concentration × fractional oxygen saturation) may provide valuable information. We hypothesized that OxyHem predicts survival of LTx recipients at the onset of CLAD. Data from 292 LTx recipients with CLAD were analyzed. After excluding patients with missing data or supplemental oxygen, the final population comprised 218 patients. The relationship between survival upon CLAD and OxyHem was analyzed by Cox regression analyses and ROC curves. Among the 218 patients (102 males, 116 females), 128 (58.7%) died, median survival time after CLAD onset being 1,156 days. Survival was significantly associated with type of transplantation, time to CLAD, CLAD stage at onset, and OxyHem, which was superior to Hb or oxygen saturation. The risk for death after CLAD increased by 14% per reduction of OxyHem by 1 g/dL, and values below 11 g/dL corresponded to an 80% increase in mortality risk. Thus, OxyHem was identified as an independent predictor of mortality after CLAD onset. Whether it is useful in supporting therapeutic decisions and potentially home monitoring in the surveillance of lung transplant recipients has to be studied further.


Melatonin as an anti-inflammatory agent in radiotherapy.
Radiotherapy is one of the most relevant treatment options for cancer therapy with or without other treatment modalities including immunotherapy, surgery and chemotherapy. Exposure to heavy doses of ionizing radiation during radiotherapy results in short and long term side effects. It appears that many of these side effects are linked to inflammatory responses during treatment or after prolonged use. Inflammation is mediated by various genes and cytokines related to immune system responses caused by massive cell death following radiotherapy. This phenomenon is more obvious, particularly after exposure to clinical doses of radiotherapy. Inflammation is involved in the amplification of acute responses, genomic instability and also long term pathological changes in normal tissues. Moreover, inflammation attenuates responses of the tumor to radiotherapy through some mechanisms such as angiogenesis. Thus, the management of inflammation is one of the most interesting aims in cancer radiotherapy. Melatonin, known as a natural product in the body, has been of much interest for its anti-inflammatory properties. Some studies have proposed melatonin as a novel anti-inflammation agent. This literature review will concentrate on the anti-inflammatory properties of melatonin that may help the management of different inflammatory signaling pathways in both tumor and normal tissues.


New subgenera and a new species of the genus Raphignathus Dugès (Prostigmata, Raphignathidae), with taxonomic notes on the genus Neoraphignathus Smiley & Moser.
Four new subgenera in the genus Raphignathus Dugès are hereby proposed: Raphignathus (Raphignathus), subgen. nov., Raphignathus (Monoraphignathus), subgen. nov., Raphignathus (Diraphignathus), subgen. nov., and Raphignathus (Triraphignathus), subgen. nov. These subgenera are diagnosed by the number of setae on the interscutal membrane of females. A new species, R. (D.) neohecmatanaensissp. nov., is described and illustrated based on females collected from Ziziphusspina-christi Mill. (Rhamnaceae). The taxonomic status of the monotypic genus Neoraphignathus Smiley & Moser and three species (R.evidus, R.hsiufui, and R.johnstoni) are discussed. A key to world species of the family Raphignathidae is given.


Antibacterial Activity, in Vitro Cytotoxicity, and Cell Cycle Arrest of Gemini Quaternary Ammonium Surfactants.
Twelve gemini quaternary ammonium surfactants have been employed to evaluate the antibacterial activity and in vitro cytotoxicity. The antibacterial effects of the gemini surfactants are performed on Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) with minimum inhibitory concentrations (MIC) ranging from 2.8 to 167.7 μM. Scanning electron microscopy (SEM) analysis results show that these surfactants interact with the bacterial cell membrane, disrupt the integrity of the membrane, and consequently kill the bacteria. The data recorded on C6 glioma and HEK293 human kidney cell lines using an MTT assay exhibit low half inhibitory concentrations (IC50). The influences of the gemini surfactants on the cell morphology, the cell migration ability, and the cell cycle are observed through hematoxylin-eosin (HE) staining, cell wound healing assay, and flow cytometric analyses, respectively. Both the values of MIC and IC50 decrease against the growth of the alkyl chain length of the gemini surfactants with the same spacer group. In the case of surfactants 12-s-12, the MICs and IC50s are found to decrease slightly with the spacer chain length changing from 2 to 8 and again to increase at higher spacer length (s = 10-12). All of the gemini surfactants show great antibacterial activity and cytotoxicity, and they might exhibit potential applications in medical fields.


Pharmacogenetic principles in the Hippocratic writings.
Twenty-five centuries after Hippocrates, we are at the beginning of an era of individualized, molecular, and genomic medicine, where diagnosis, prognosis, and treatment will be increasingly based on our understanding of the human organism's genetic and molecular composition. The most difficult task in this process has proven to be the application of the scientific advancements to the clinical setting, and clinical pharmacology is an essential discipline in this translational process. This article traces the roots of the emerging personalized and genome-based medicine to the teachings of Hippocrates.


[Calnexin (CNX) enhances the killing effect of CD8+ T cells on colorectal cancer cells by promoting MHC I expression].
Objective To investigate the killing effect and molecular mechanism of aberrant expression of calnexin (CNX) in the colorectal cancer (CRC) on the CD8+ T immune cells. Methods Immunohistochemistry was used to detect CNX protein level in 102 pairs of CRC cancer and adjacent non-cancerous tissues. Western blotting was employed to examine the protein expression of MHC I in the HCT-15 cells overexpressed with CNX or in the SW480 cells whose CNX expressions were knockdown by siRNA. Murine CD8+ T cells isolated from the spleen were cocultured with CT-26 murine CRC cells infected with lentivirus-mediated CNX overexpression. The killing effect of CD8+ T cells on CT-26 cells was determined by cytotoxicity kit. The secretion of interferon γ (IFN-γ) and tumor necrosis factor α (TNF-α) in the culture medium were examined by ELISA. Results The protein level of CNX in colorectal cancer tissues were significantly lower than that in non-cancerous tissues. CNX overexpressed in HCT-15 cells was upregulated and CNX knockdown in SW480 cells downregulated the MHC I expression in these cells. Furthermore, the overexpression of CNX could not only enhance the killing effect of CD8+ T cells on CT-26 cells, but also promote the secretion of IFN-γ and TNF-α from these cells. Conclusion CNX can enhance the killing potential of CD8+ T cells on tumor cells through upregulating the MHC I expression in colorectal cancer cells.


[Continuous blood purification therapy on 16 patients with diabetic ketoacidosis and acute kidney injury].
To determine the effectiveness of Continuous blood purification (CBP) therapy on diabetic ketoacidosis (DKA) and acute kidney injury (AKI) in diabetic nephropathy (DN) patients. Sixteen DN patients who developed severe DKA and AKI between 2008 and 2011 in the West China Hospital were recruited. All of the recruited patients presented with severe metabolic acidosis, electrolyte disturbance and dehydration. In addition to routine treatments, continuous venovenous hemofiltration (CVVH) was performed for at least 48 h with Baxter Accura or B. Braun Diapact CRRT machine and B. Braun Diacap Acute M hemofilter. Hemofiltration was accomplished using predilution bicarbonate replacement fluid at the rate of 3000 mL/h and citrate or low-molecular weight heparin (LMWH) for anticoagulation, with blood flow rates of 180 to 250 mL/min. One patient died unexpectedly 10 h after admission to hospital. The other fifteen patients had significant improvements in metabolic acidosis index after 12 hours of CVVH therapy, such as an average increase of 7.21 +/- 0.07 carbon dioxide combining power (CO2CP)and improvement of arterial PH. The blood urea nitrogen (BUN), serum creatinine (SCr), serum glucose (Glu), serum potassium (K(+)) and bloodosmotic pressure of the 15 patients decreased significantly after 48 hours of CVVH therapy. Eleven cases entered into diuretic phase and had renal functions recovered (12 +/- 5) d and (18 +/- 12) d after admission to the hospital, respectively. CVVH therapy as an early intervention can bring significant benefits to DN patients with DKA and AKI. Early institution of CVVH therapy may be considered not only for treating uremia and fluid retention but also for correcting metabolic abnormalities like metabolic acidosis.


Head injuries--what happens to survivors.
The head injury syndrome is a series of psychological characteristics observed in survivors of traumatic head injuries referred to a hospital-based rehabilitation centre. Severe cases of head injury syndrome are readily identifiable as cases of organic dementia. In less florid cases patients easily pass for normal in routine consultation situations, but upon expert psychometric assessment, elements of the head injury syndrome can be demonstrated. Such patients exhibit a high propensity to fail to live up to the normal expectations of their work and domestic environment, and suffer repeated failures and social difficulties which are the direct result of the head injury syndrome. This leads to breakdown of work and family life, increasing frustration, loss of confidence, and final disintegration of the affected personality.


Does the combination of hyperkalemia and KATP activation determine excitation rate gradient and electrical failure in the globally ischemic fibrillating heart?
Ventricular fibrillation (VF) in the globally ischemic heart is characterized by a progressive electrical depression manifested as a decline in the VF excitation rate (VFR) and loss of excitability, which occur first in the subepicardium (Epi) and spread to the subendocardium (Endo). Early electrical failure is detrimental to successful defibrillation and resuscitation during cardiac arrest. Hyperkalemia and/or the activation of ATP-sensitive K(+) (KATP) channels have been implicated in electrical failure, but the role of these factors in ischemic VF is poorly understood. We determined the VFR-extracellular K(+) concentration ([K(+)]o) relationship in the Endo and Epi of the left ventricle during VF in globally ischemic hearts (Isch group) and normoxic hearts subjected to hyperkalemia (HighK group) or a combination of hyperkalemia and the KATP channel opener cromakalim (HighK-Crom group). In the Isch group, Endo and Epi values of [K(+)]o and VFR were compared in the early (0-6 min), middle (7-13 min), and late (14-20 min) phases of ischemic VF. A significant transmural gradient in VFR (Endo > Epi) was observed in all three phases, whereas a significant transmural gradient in [K(+)]o (Epi > Endo) occurred only in the late phase of ischemic VF. In the Isch group, the VFR decrease and inexcitability started to occur at much lower [K(+)]o than in the HighK group, especially in the Epi. Combining KATP activation with hyperkalemia only shifted the VFR-[K(+)]o curve upward (an effect opposite to real ischemia) without changing the [K(+)]o threshold for asystole. We conclude that hyperkalemia and/or KATP activation cannot adequately explain the heterogeneous electrical depression and electrical failure during ischemic VF.


The ALK1‑Smad1/5‑ID1 pathway participates in tumour angiogenesis induced by low‑dose photodynamic therapy.
Photodynamic therapy (PDT) is an effective and low‑invasive tumour therapy. However, it can induce tumour angiogenesis, which is a main factor leading to tumour recurrence and metastasis. Activin receptor‑like kinase‑1 (ALK1) is a key factor regulating angiogenesis. However, it remains unclear whether ALK1 plays an unusual role in low‑dose PDT‑induced tumour angiogenesis. In the present study, human umbilical vein endothelial cells (HUVECs) co‑cultured with breast cancer MDA‑MB‑231 cells (termed HU‑231 cells) were used to construct an experimental model of tumour angiogenesis induced by low‑dose PDT. The viability, and the proliferative, invasive, migratory, as well as the tube‑forming ability of the HU‑231 cells were evaluated following low‑dose PDT. In particular, ALK1 inhibitor and and an adenovirus against ALK1 were used to further verify the role of ALK1 in low‑dose PDT‑induced tumour angiogenesis. Moreover, the expression of ALK1, inhibitor of DNA binding 1 (ID1), Smad 1, p‑Smad1/5, AKT and PI3K were detected in order to verify the underlying mechanisms. The findings indicated that low‑dose PDT enhanced the proliferative ability of the HU‑231 cells and reinforced their migratory, invasive and tube formation capacity. However, these effects were reversed with the addition of an ALK1 inhibitor or by the knockdown of ALK1 using adenovirus. These results indicated that ALK1 was involved and played a critical role in tumour angiogenesis induced by low‑dose PDT. Furthermore, ALK1 was found to participate in PDT‑induced tumour angiogenesis by activating the Smad1/5‑ID1 pathway, as opposed to the PI3K/AKT pathway. On the whole, the present study, for the first time, to the best of our knowledge, demonstrates that ALK1 is involved in PDT‑induced tumour angiogenesis. The inhibition of ALK1 can suppress PDT‑induced tumour angiogenesis, which can enhance the effects of PDT and may thus provide a novel treatment strategy for PDT.


Has the diversity of clinical and biological manifestations of systemic lupus erythematosus a correspondent in the diversity of immune mechanisms? Observations based on a Rowell's syndrome case associated with arthritis and nephritis.
This paper draws attention to the relationship between the clinical and biological picture of SLE and the immune mechanisms of this disease. The presence, in the same patient, of erythema multiforme-like skin lesions and erythemato-squamous lesions specific for SLE together with a characteristic immune picture (speckled antinuclear antibodies (ANAs), positive anti-Ro antibodies, positive rheumatoid factor) raise the question of a relationship between the immune mechanisms in SLE and the clinical picture. A case of Rowell's syndrome is discussed: systemic lupus erythematosus diagnosed on the occasion of an erythema multiforme-like rash. Starting from this case, we analyse if the clinical and biological picture in SLE is an expression of the immune mechanisms involved in this disease. Our patient presented with speckled antinuclear antibodies, positive rheumatoid factor, anti-Ro antibodies, suggestive of Rowell's syndrome. The patient manifested rheumatoid-like articular pain and high titer rheumatoid factor. Clinically, we found erythema multiforme-like and erythemato-squamous lesions. The patient developed nephrotic syndrome (proteinuria 11.8g/24h), and renal failure (creatinine 3.08 mg/dl). The renal biopsy showed mesangial proliferative glomerulonephritis class II (ISN/RPS). Under treatment with prednisone the nephrotic syndrome evolved into remission (traces of proteinuria) and serum creatinine declined (1.03 mg/dl). The cutaneous syndrome had a spectacular evolution, too. The question is raised of the existence in Rowell's syndrome of immune mechanisms commonly encountered in SLE and a subset associated with the cutaneous erythema multiforme-like rash and pseudo-rheumatoid arthritis manifestations.


[Fact and figures about diabetes in Italy].
Fact and figures about diabetes in Italy. The diabetes is a true social, political, economical and health emergency that affects all Countries in the worls. The document "Facts and figures about diabetes in Italy" aims to draw a clear picture of the situation in Italy. The report presents updated information on mortality and morbility, clinical impact of diabetes on health care, costs, effective interventions for its prevention and early treatment. It and analyzes the impact of diabetes on quality of care and reports initiative, challenges and strategies to face this illness.


A comparison between end-of-life home care clients with cancer and heart failure in Ontario.
Individuals with heart failure experience complex symptoms and have a poor prognosis, comparable to those with cancer. However, people with heart failure are less likely to be recognized as needing palliative care. We compared seriously ill home care clients with cancer versus those with heart failure to explore differences between them using existing data from the Resident Assessment Instrument for Home Care (RAI-HC). The sample included 1,475 older adults (65+) in Ontario who had a prognosis of less than 6 months or severe health instability. Clients with heart failure were significantly older (p < .0001) and experienced significantly higher rates of impairment in activities of daily living (p = .005), cognitive impairment (p < .0001), and severe health instability (p < .0001), but were significantly less likely to have a prognosis of less than 6 months (p < .0001). Home care clients with heart failure have needs similar to those with cancer yet are typically not identified as having a terminal prognosis.


Population Genomic Evidence for a Repeated Introduction and Rapid Expansion of the Fungal Maize Pathogen Setosphaeria turcica in Europe.
Modern agricultural practices, climate change, and globalization foster the rapid spread of plant pathogens, such as the maize fungal pathogen Setosphaeria turcica, which causes Northern corn leaf blight and expanded into Central Europe during the twentieth century. To investigate the rapid expansion of S. turcica, we sequenced 121 isolates from Europe and Kenya. Population genomic inference revealed a single genetically diverse cluster in Kenya and three clonal lineages with low diversity, as well as one cluster of multiple clonal sublineages in Europe. Phylogenetic dating suggests that all European lineages originated through sexual reproduction outside Europe and were subsequently introgressed multiple times. Unlike isolates from Kenya, European isolates did not show sexual recombination, despite the presence of both MAT1-1 and MAT1-2 mating types. For the clonal lineages, coalescent model selection supported a selectively neutral model with strong exponential population growth, rather than models with pervasive positive selection caused by host defense resistance or environmental adaptation. Within clonal lineages, phenotypic variation in virulence to different monogenic resistances, which defines the pathogen races, suggests that these races may originate from repeated mutations in virulence genes. Association testing based on k-mers did not identify genomic regions linked to pathogen races, but it did uncover strongly differentiated genomic regions between clonal lineages, which harbor genes with putative roles in pathogenicity. In conclusion, the expansion and population growth of S. turcica in Europe are mainly driven by an expansion of the maize cultivation area and not by rapid adaptation.


Naturally Engineered Maturation of Cardiomyocytes.
Ischemic heart disease remains one of the most prominent causes of mortalities worldwide with heart transplantation being the gold-standard treatment option. However, due to the major limitations associated with heart transplants, such as an inadequate supply and heart rejection, there remains a significant clinical need for a viable cardiac regenerative therapy to restore native myocardial function. Over the course of the previous several decades, researchers have made prominent advances in the field of cardiac regeneration with the creation of in vitro human pluripotent stem cell-derived cardiomyocyte tissue engineered constructs. However, these engineered constructs exhibit a functionally immature, disorganized, fetal-like phenotype that is not equivalent physiologically to native adult cardiac tissue. Due to this major limitation, many recent studies have investigated approaches to improve pluripotent stem cell-derived cardiomyocyte maturation to close this large functionality gap between engineered and native cardiac tissue. This review integrates the natural developmental mechanisms of cardiomyocyte structural and functional maturation. The variety of ways researchers have attempted to improve cardiomyocyte maturation in vitro by mimicking natural development, known as natural engineering, is readily discussed. The main focus of this review involves the synergistic role of electrical and mechanical stimulation, extracellular matrix interactions, and non-cardiomyocyte interactions in facilitating cardiomyocyte maturation. Overall, even with these current natural engineering approaches, pluripotent stem cell-derived cardiomyocytes within three-dimensional engineered heart tissue still remain mostly within the early to late fetal stages of cardiomyocyte maturity. Therefore, although the end goal is to achieve adult phenotypic maturity, more emphasis must be placed on elucidating how the in vivo fetal microenvironment drives cardiomyocyte maturation. This information can then be utilized to develop natural engineering approaches that can emulate this fetal microenvironment and thus make prominent progress in pluripotent stem cell-derived maturity toward a more clinically relevant model for cardiac regeneration.


Characterization of Lipid Alterations by Oncogenic PIK3CA Mutations Using Untargeted Lipidomics in Breast Cancer.
Lipids play crucial biological roles in health and disease, including in cancers. The phosphatidylinositol 3-kinase (PI3K) signaling pathway is a pivotal promoter of cell growth and proliferation in various types of cancer. The somatic mutations in PIK3CA, the gene coding for the catalytic subunit p110α of PI3K, are frequently present in cancer cells, including breast cancer. Although the most prominent mutants, represented by single amino acid substitutions in the helical domain in exon 9 (E545K) and the kinase domain in exon 20 (H1047R) are known to cause a gain of PI3K function, activate AKT signaling and induce oncogenic transformation, the effect of these mutations on cellular lipid profiles has not been studied. We carried out untargeted lipidomics using liquid chromatography-tandem mass spectrometry to detect the lipid alterations in mammary gland epithelial MCF10A cells with isogenic knockin of these mutations. A total of 536 species of lipids were analyzed. We found that the levels of monosialogangliosides, signaling molecules known to enhance cell motility through PI3K/AKT pathway, were significantly higher in both mutants. In addition, triglycerides and ceramides, lipid molecules known to be involved in promoting lipid droplet production, cancer cell migration and invasion, were increased, whereas lysophosphatidylcholines and phosphatidylcholines that are known to inhibit cancer cell motility were decreased in both mutants. Our results provide novel insights into a potential link between altered lipid profile and carcinogenesis caused by the PIK3CA hotspot mutations. In addition, we suggest untargeted lipidomics offers prospects for precision/personalized medicine by unpacking new molecular substrates of cancer biology.


In vivo models of developmental toxicology.
The founding principles of teratology/developmental toxicology state that a developmental toxicants cause dysmorphogenesis when conceptuses are exposed at a sufficient dosage during a sensitive period of development in a sensitive species. While in vitro approaches in developmental toxicology can provide a means to assess the potency of toxicants, ultimately, the need to use whole animal models to demonstrate embryotoxicity is necessary to fully extrapolate findings to the human condition. This chapter is dedicated to reviewing the advantages of specific animal models and how these animal models may be used to assess toxicity in the embryo, both descriptively and mechanistically.


Bridging length and time scales in sheared demixing systems: from the Cahn-Hilliard to the Doi-Ohta model.
We develop a systematic coarse-graining procedure which establishes the connection between models of mixtures of immiscible fluids at different length and time scales. We start from the Cahn-Hilliard model of spinodal decomposition in a binary fluid mixture under flow from which we derive the coarse-grained description. The crucial step in this procedure is to identify the relevant coarse-grained variables and find the appropriate mapping which expresses them in terms of the more microscopic variables. In order to capture the physics of the Doi-Ohta level, we introduce the interfacial width as an additional variable at that level. In this way, we account for the stretching of the interface under flow and derive analytically the convective behavior of the relevant coarse-grained variables, which in the long wavelength limit recovers the familiar phenomenological Doi-Ohta model. In addition, we obtain the expression for the interfacial tension in terms of the Cahn-Hilliard parameters as a direct result of the developed coarse-graining procedure. Finally, by analyzing the numerical results obtained from the simulations on the Cahn-Hilliard level, we discuss that dissipative processes at the Doi-Ohta level are of the same origin as in the Cahn-Hilliard model. The way to estimate the interface relaxation times of the Doi-Ohta model from the underlying morphology dynamics simulated at the Cahn-Hilliard level is established.


[The clinical analysis of 51 patients with thoracic aortic dissection].
To summarize the clinical experience about surgical treatment of aortic dissection. The clinical data of 51 patients with aortic dissection admitted from December 2004 to December 2006 were analyzed retrospectively. There were 35 male and 16 female patients with a mean age of 55.7 years (ranged from 18 to 83-years-old). Twenty-seven patients of type I was performed under deep hypothermic circulatory arrest and selected cerebral perfusion with stent-graft which was implanted into the descending aorta through aorta arch. Five patients of type II was performed including Bentall operation in 3 patients, Wheat operation in 1 patient, ascending aorta replacement in 1 patient. Nineteen patients of type III was performed with stent-graft which was implanted into the descending aorta through aorta arch under deep hypothermic circulatory arrest. The time of cardiopulmonary bypass (CPB) in type I patients was 250 to 290 min with an average of (274 +/- 53) min, and the arrest time was 40 to 59 min with an average of (53 +/- 14) min. CPB time of type II patients was 130 to 159 min with an average of (146 +/- 43) min, and the cross clamp time was 60 to 79 min with an average of (66 +/- 15) min. CPB time of type III patients was 240 to 280 min with an average of (260 +/- 28) min, and the arrest time was 20 to 27 min with an average of (24 +/- 3) min. The mean hemorrhage volume of the entire group was (500 +/- 250) ml. The mean ICU retention time was (5.0 +/- 1.5) d and the length of stay was (15.0 +/- 2.5) d. Three patients died during perioperative period. Two patients appeared cerebrovascular accident after operation. One patient appeared descending aorta dilation in the follow-up of 2 to 21 months. Different clinical manifestations and treatment should be selected according to the different condition of aortic dissection aneurysm.


[The effect of air pollution on man].
During the last years, the human organism has been exposed to airborne pollutants to an increasing extend, among them the following most important: SO2, NOx , hydrocarbons, CO, photochemical smog, airborne particles containing the heavy metals lead and cadmium. The local effects on the lung and the bronchial mucous membrane are described including irritations of the mucous membrane and disturbances of the defence and elimination mechanisms. The main consequences are chronic bronchitis and bronchial carcinoma. The systemic effects of inhaled airborne pollutants are mutagenicity, cancerogenicity and toxicity. Disturbances of the immunological system may occur. Details as regard the effects of carbon monoxide, lead and cadmium, photochemical smog and SO2 are given.


[Venous malformations: clinical characteristics and differential diagnosis].
Venous malformations (VM) are localized defects of blood vessels that are due to vascular dysmorphogenesis. These slow-flow lesions can affect any tissue or organ. Clinically, a cutaneous VM is characterized by a bluish mass that is compressible on palpation. Phleboliths are commonly present. Symptoms depend on location and size. VM are often sporadic and isolated, however, they can be associated with other malformations and be part of a syndrome; Klippel-Trenaunay (capillary-lymphatico-venous malformation with limb hypertrophy) is the most common. Glomuvenous malformation (GVM) is another type of venous anomaly. In contrast to VM, GVM is often painful on palpation and not compressible. Clinical diagnosis of VM is often made in the presence of a bluish cutaneous lesion: however, other lesions can mimick VM. The most frequent anomalies are a blue naevus, a hemorrhagic lymphatic malformation, a sub-cutaneous hemangioma or even the presence of dilated superficial normal veins due to underlying venous stenoses. This chapter will detail the clinical characteristics of venous anomalies and their differential diagnosis.


MicroRNA-885-5p promotes osteosarcoma proliferation and migration by downregulation of cell division cycle protein 73 homolog expression.
Osteosarcoma (OS) is the most common primary malignant bone tumor. Numerous studies have strongly implicated the ectopic expression of microRNAs (miRNAs/miRs), including miR-885-5p, which is aberrantly expressed in several cancer types, in multiple cancer-related processes. However, the role of miR-885-5p in OS remains unknown. In the present study, it was found that the expression of miR-885-5p was markedly upregulated in OS cell lines and clinical tissues. Moreover, high expression of miR-885-5p was significantly associated with the development of OS. The human OS MG-63 cell line was transfected with recombinant lentivirus to regulate miR-885-5p expression. Overexpressed miR-885-5p significantly promoted the proliferation and migration of MG-63 cells in vitro, while downregulating miR-885-5p expression reversed these effects. Furthermore, bioinformatic analysis was used to predict the potential target genes of miR-885-5p, and cell division cycle protein 73 homolog (CDC73) was identified as a novel and direct target of miR-885-5p. This interaction was further confirmed using reverse transcription-quantitative polymerase chain reaction, western blotting and luciferase activity assays. These findings suggest that miR-885-5p serves a critical role in facilitating OS proliferation and migration, and can regulate CDC73 expression in OS cells and tissues. Thus, miR-885-5p could be a promising novel therapeutic biomarker for OS.


Nasal-Swab Testing Misses Patients with Low SARS-CoV-2 Viral Loads.
The urgent need for large-scale diagnostic testing for SARS-CoV-2 has prompted pursuit of sample-collection methods of sufficient sensitivity to replace sampling of the nasopharynx (NP). Among these alternatives is collection of nasal-swab samples, which can be performed by the patient, avoiding the need for healthcare personnel and personal protective equipment. Previous studies have reached opposing conclusions regarding whether nasal sampling is concordant or discordant with NP. To resolve this disagreement, we compared nasal and NP specimens collected by healthcare workers in a cohort consisting of individuals clinically suspected of COVID-19 and outpatients known to be SARS-CoV-2 RT-PCR positive undergoing follow-up. We investigated three different transport conditions, including traditional viral transport media (VTM) and dry swabs, for each of two different nasal-swab collection protocols on a total of 308 study participants, and compared categorical results and Ct values to those from standard NP swabs collected at the same time from the same patients. All testing was performed by RT-PCR on the Abbott SARS-CoV-2 RealTime EUA (limit of detection [LoD], 100 copies viral genomic RNA/mL transport medium). We found high concordance (Cohen's kappa >0.8) only for patients with viral loads above 1,000 copies/mL. Those with viral loads below 1,000 copies/mL, the majority in our cohort, exhibited low concordance (Cohen's kappa = 0.49); most of these would have been missed by nasal testing alone. Previous reports of high concordance may have resulted from use of assays with higher LoD (≥1,000 copies/mL). These findings counsel caution in use of nasal testing in healthcare settings and contact-tracing efforts, as opposed to screening of asymptomatic, low-prevalence, low-risk populations. Nasal testing is an adjunct, not a replacement, for NP.