Coupling I-125 permanent implant prostate brachytherapy Monte Carlo dose calculations with radiobiological models.
To investigate the coupling of radiobiological models with patient-specific Monte Carlo (MC) dose calculations for permanent implant prostate brachytherapy (PIPB). To compare radiobiological indices evaluated with different radiobiological models using MC and simulated AAPM TG-43 dose calculations. Three-dimensional dose distributions previously computed using MC techniques with two types of patient models, TG43sim (AAPM TG-43 water-based conditions) and MCDmm (realistic tissues and interseed effects), for 613 PIPB patients are coupled with biological dose and tumour control probability (TCP) models. Two approaches and their extensions are considered to evaluate biological doses, biologically effective dose (BED) and isoeffective dose (IED), as well as two methods to evaluate TCP. Three novel extensions of equivalent uniform biologically effective dose (EUBED) are suggested which consider the spatial distribution of doses within the target volume. Adopted radiobiological model parameter values (α, β, etc) are those suggested by AAPM TG-137, and sensitivity to parameter choice is discussed. MCDmm dose calculations can reveal low doses in the prostate target volume, due to tissue heterogeneities or inter-seed effects; considering these low doses in EUBED calculations can lower TCP estimates by up to 70%, with largest differences in patients with calcifications. There are large variations in biological doses and TCPs evaluated over the 613 patient cohort for each radiobiological model considered, reflecting the spectrum of physical doses calculated for these patients with either MCDmm or TG43sim. Depending on the model details, BED, IED and EUBED are, on average, 6.0-9.8%, 7.4-9.2% and 1.8-15% higher, respectively, with TG43sim than MCDmm. TCP estimates computed using MCDmm dose distributions are much lower than expected based on past treatment outcome studies, suggesting a need to re-assess model parameters when evaluating radiobiological indices coupled with heterogeneous tissue model-based dose calculations. Cohort average differences in biological dose and TCP estimates between radiobiological models are generally larger than differences for any one radiobiological model evaluated with TG43sim or MCDmm dose calculations. However, heterogeneous tissue dose calculations, like MCDmm, can identify clinically-relevant low dose volumes, e.g., in patients with calcifications, which would otherwise be missed with TG-43. In addition to affecting physical dose distributions, these low dose volumes can largely impact radiobiological dose and TCP estimates, which further motivates the clinical implementation of model-based dose calculations for PIPB.


A simple and rapid external standard calibration HPLC method for determination of lumefantrine in dried blood spot samples from malaria patients in Botswana.
A simple external calibration liquid chromatography-diode array detector method was developed, validated, and applied for the determination of lumefantrine (Lum) in dried blood spot (DBS) samples collected from malaria patients in Botswana. The samples were validated in accordance with the United States Food and Drug Administration guidelines for bioanalytical methods after sample preparation using solid-liquid extraction. Separation was achieved using an XTerra C18 column (50 × 4.6 mm, 5 μm), and a binary solvent system of acetonitrile and water adjusted to pH 2.3 was used as the mobile phase. The validated method was applied for the determination of Lum in DBS samples collected from malaria patients infected with Plasmodium falciparum in Botswana. The calibration curve was linear between 0.5 and 12 μg/mL with a coefficient of determination (R2 ) of 0.9996. The limit of detection and the lower limit of quantification were 0.5 and 1.4 μg/mL, respectively. The efficiency of extraction measured as percentage recovery ranged between 84.2% and 107.8% at the three quality control (QC) levels, that is, low QC, mid QC, and high QC. In conclusion, data suggest that the method is suitable for the determination of trace Lum in biofluids and can also be used for therapeutic drug monitoring and pharmacokinetic profiling.


Bacterial guanine nucleotide exchange factors SopE-like and WxxxE effectors.
Subversion of Rho family small GTPases, which control actin dynamics, is a common infection strategy used by bacterial pathogens. In particular, Salmonella enterica serovar Typhimurium, Shigella flexneri, enteropathogenic Escherichia coli (EPEC), and enterohemorrhagic Escherichia coli (EHEC) translocate type III secretion system (T3SS) effector proteins to modulate the Rho GTPases RhoA, Cdc42, and Rac1, which trigger formation of stress fibers, filopodia, and lamellipodia/ruffles, respectively. The Salmonella effector SopE is a guanine nucleotide exchange factor (GEF) that activates Rac1 and Cdc42, which induce "the trigger mechanism of cell entry." Based on a conserved Trp-xxx-Glu motif, the T3SS effector proteins IpgB1 and IpgB2 of Shigella, SifA and SifB of Salmonella, and Map of EPEC and EHEC were grouped together into a WxxxE family; recent studies identified the T3SS EPEC and EHEC effectors EspM and EspT as new family members. Recent structural and functional studies have shown that representatives of the WxxxE effectors share with SopE a 3-D fold and GEF activity. In this minireview, we summarize contemporary findings related to the SopE and WxxxE GEFs in the context of their role in subverting general host cell signaling pathways and infection.


Feasibility Study of a Time-of-Flight Brain Positron Emission Tomography Employing Individual Channel Readout Electronics.
The purpose of this study was to investigate the feasibility of a time-of-flight (TOF) brain positron emission tomography (PET) providing high-quality images. It consisted of 30 detector blocks arranged in a ring with a diameter of 257 mm and an axial field of view of 52.2 mm. Each detector block was composed of two detector modules and two application-specific integrated circuit (ASIC) chips. The detector module was composed of an 8 × 8 array of 3 × 3 mm2 multi-pixel photon counters and an 8 × 8 array of 3.11 × 3.11 × 15 mm3 lutetium yttrium oxyorthosilicate scintillators. The 64-channel individual readout ASIC was used to acquire the position, energy, and time information of a detected gamma ray. A coincidence timing resolution of 187 ps full width at half maximum (FWHM) was achieved using a pair of channels of two detector modules. The energy resolution and spatial resolution were 6.6 ± 0.6% FWHM (without energy nonlinearity correction) and 2.5 mm FWHM, respectively. The results of this study demonstrate that the developed TOF brain PET could provide excellent performance, allowing for a reduction in radiation dose or scanning time for brain imaging due to improved sensitivity and signal-to-noise ratio.


A global method for handling fluorescence spectra at high concentration derived from the competition between emission and absorption of colloidal CdTe quantum dots.
We investigate the effects of the concentration of CdTe quantum dots (QDs) on their fluorescence in water. The emission spectra, acquired in right angle geometry, exhibit highly variable shapes. The measurements evidence a critical value of the concentration beyond which the intensity and the spectral bandwidth decrease and the fluorescence maximum is redshifted. To account for these observations, we develop a model based on the primary and secondary inner filter effects. The accuracy of the model ensures that the concentration dependent behaviour of QD fluorescence is completely due to inner filter effects. This result is all the more interesting because it precludes the assumption of dynamic quenching. As a matter of fact, the decrease of the emission intensity is not a consequence of collisional quenching but an effect of competition between fluorescence and absorption. We even show that this phenomenon is linked not only to the QD concentration but also to the geometric configuration of the setup. Hence, our analytical model can be easily adapted to every fluorescence spectroscopy configuration to quantitatively predict or correct inner filter effects in the case of QDs or any fluorophore, and thus improve the handling of fluorescence spectroscopy for highly concentrated solutions.


Enhanced expressions and activations of leukotriene C4 synthesis enzymes in D-galactosamine/lipopolysaccharide-induced rat fulminant hepatic failure model.
To investigate the expression and activity of leukotriene C4 (LTC4) synthesis enzymes and their underlying relationship with cysteinyl leukotriene (cys-LT) generation in a rat fulminant hepatic failure (FHF) model induced by D-galactosamine/lipopolysaccharide (D-GalN/ LPS). Rats were treated with D-GalN (300 mg/kg) plus LPS (0.1 mg/kg) for 1, 3, 6, and 12 h. Enzyme immunoassay was used to determine the hepatic cys-LT content. Reverse transcription-polymerase chain reaction (RT-PCR), Western blot or immunohistochemical assay were employed to assess the expression or location of LTC4 synthesis enzymes, which belong to membrane associated proteins in eicosanoid and glutathione (MAPEG) metabolism superfamily. Activity of LTC4 synthesis enzymes was evaluated by determination of the products of LTA4 after incubation with liver microsomes using high performance liquid chromatography (HPLC). Livers were injured after treatment with D-GalN/LPS, accompanied by cys-LT accumulation at the prophase of liver injury. Both LTC4 synthase (LTC4S) and microsomal glutathione-S-transferase (mGST) 2 were expressed in the rat liver, while the latter was specifically located in hepatocytes. Their mRNA and protein expressions were up-regulated at an earlier phase after treatment with D-GalN/LPS. Meantime, a higher activity of LTC4 synthesis enzymes was detected, although the activity of LTC4S played the main role in this case. The expression and activity of both LTC4S and mGST2 are up regulated in a rat FHF model, which are, at least, partly responsible for cys-LT hepatic accumulation.


Distinguishing Emission-Associated Ambient Air PM2.5 Concentrations and Meteorological Factor-Induced Fluctuations.
Although PM2.5 (particulate matter with aerodynamic diameters less than 2.5 μm) in the air originates from emissions, its concentrations are often affected by confounding meteorological effects. Therefore, direct comparisons of PM2.5 concentrations made across two periods, which are commonly used by environmental protection administrations to measure the effectiveness of mitigation efforts, can be misleading. Here, we developed a two-step method to distinguish the significance of emissions and meteorological factors and assess the effectiveness of emission mitigation efforts. We modeled ambient PM2.5 concentrations from 1980 to 2014 based on three conditional scenarios: realistic conditions, fixed emissions, and fixed meteorology. The differences found between the model outputs were analyzed to quantify the relative contributions of emissions and meteorological factors. Emission-related gridded PM2.5 concentrations excluding the meteorological effects were predicted using multivariate regression models, whereas meteorological confounding effects on PM2.5 fluctuations were characterized by probabilistic functions. When the regression models and probabilistic functions were combined, fluctuations in the PM2.5 concentrations induced by emissions and meteorological factors were quantified for all model grid cells and regions. The method was then applied to assess the historical and future trends of PM2.5 concentrations and potential fluctuations on global, national, and city scales. The proposed method may thus be used to assess the effectiveness of mitigation actions.


Discolored ureteral stents: findings in urinalysis and urine culture.
Discolored ureteral stents are sometimes encountered in daily clinical practice; however, the mechanism(s) underlying the development of discolored ureteral stents remain unknown. In this study, we retrospectively analyzed the characteristics of discolored ureteral stents based on the results of a urinalysis and urine culture. We identified a total of 26 patients with discolored ureteral stents and compared the findings in the urinalyses and urine culture in 21 discolored versus 45 non-colored ureteral stents. The median and mean (± SD) duration of stenting time was 78.0 and 81.3 (± 21.3) days for the discolored ureteral stents and 69.0 and 74.9 (± 19.8) days for the non-colored ureteral stents, respectively (P = 0.25). The discolored ureteral stents were associated with a higher mean urine pH than the non-colored ureteral stents (mean: 6.4 vs 6.0, P< 0.05). There were no significant differences between the two groups in the RBC (P = 0.51) and WBC (P = 0.35) counts in the urinalyses. In addition, the rate of a positive culture in the patients with discolored stents [20 of 21 (95.2%)] was significantly (P <0.01) higher than that observed in the patients with non-colored ureteral stents [33 of 45 (73.3%)]. In this study, the subjects with discolored ureteral stents showed a significantly higher likelihood of having a positive urine culture and also demonstrated higher pH values in the urinalyses. However, no clear cut-off point to predict discoloration was indicated.


Microsurgical epididymovasostomy by loop intussusception. A new technique in the rat model.
A new technique, epididymal loop intussusception into the vas deferens, is offered as an improved form compared with the side-to-end method. Loop intussusception has been experimentally studied bilaterally in 15 adult male rats. This report provides details of the technique. A 3-month anatomical and histological follow-up showed a 100% patency rate, no anastomotic stricture, and reabsorption of the blind intussuscepted loop in all but 3 cases. Macroscopic sperm granulomas were present in only 10% of the anastomoses examined. Advantages and disadvantages of microsurgical epididymovasostomy techniques are discussed.


Central Role of Protein Kinase A in Promoting Trigeminal Nociception in an In Vivo Model of Temporomandibular Disorders.
To investigate cellular changes in the spinal trigeminal nucleus (STN) and trigeminal ganglion (TG) associated with trigeminal nociception mediated by inflammation in the temporomandibular joint (TMJ). Male Sprague-Dawley rats (n = 86) were utilized to investigate cellular and behavioral responses to prolonged TMJ inflammation caused by bilateral injection of Complete Freund's Adjuvant (CFA) in the TMJ capsules. To investigate the cellular effects of protein kinase A (PKA) in the STN, rats were injected intrathecally with the selective PKA inhibitor KT5720 prior to injection of CFA into both TMJ capsules. Levels of calcitonin gene-related peptide (CGRP), active PKA, and ionized calcium-binding adapter molecule 1 (Iba1) in the STN and expression of phosphorylated extracellular regulated kinases (p-ERK) in the TG were determined with immunohistochemistry (n ≥ 3 experiments per test condition). Nocifensive head withdrawal responses to mechanical stimulation of the cutaneous tissue over the TMJ were monitored following CFA injection in the absence or presence of KT5720 (n = 7). Statistical analysis was performed using parametric analysis of variance (ANOVA) tests. Intrathecal injection of KT5720 significantly inhibited the stimulatory effect of CFA on levels of CGRP, PKA, and Iba1 in the STN. In addition, administration of KT5720 decreased the average number of CFA-induced nocifensive withdrawal responses to mechanical stimulation and the CFA-mediated increase in p-ERK expression in the ganglion. These findings provide evidence that elevated PKA activity in the STN promotes cellular events temporally associated with trigeminal nociception caused by prolonged TMJ inflammation.


Separation of drug stereoisomers by the formation of beta-cyclodextrin inclusion complexes.
For many drugs, only racemic mixtures are available for clinical use. Because different stereoisomers of drugs often cause different physiological responses, the use of pure isomers could elicit more exact therapeutic effects. Differential complexation of a variety of drug stereoisomers by immobilized beta-cyclodextrin was investigated. Chiral recognition and racemic resolution were observed with a number of compounds from such clinically useful classes as beta-blockers, calcium-channel blockers, sedative hypnotics, antihistamines, anticonvulsants, diuretics, and synthetic opiates. Separation of the diastereomers of the cardioactive and antimalarial cinchona alkaloids and of two antiestrogens was demonstrated as well. Three dimensional projections of beta-cyclodextrin complexes of propanolol, which is resolved by this technique, and warfarin, which is not, are compared. These studies have improved the understanding and application of the chiral interactions of beta-cyclodextrin, and they have demonstrated a means to measure optical purity and to isolate or produce pure enantiomers of drugs. In addition, this highly specific technique could also be used in the pharmacological evaluation of enantiomeric drugs.


The structure of allophycocyanin B from Synechocystis PCC 6803 reveals the structural basis for the extreme redshift of the terminal emitter in phycobilisomes.
Allophycocyanin B (AP-B) is one of the two terminal emitters in phycobilisomes, the unique light-harvesting complexes of cyanobacteria and red algae. Its low excitation-energy level and the correspondingly redshifted absorption and fluorescence emission play an important role in funnelling excitation energy from the hundreds of chromophores of the extramembraneous phycobilisome to the reaction centres within the photosynthetic membrane. In the absence of crystal structures of these low-abundance terminal emitters, the molecular basis for the extreme redshift and directional energy transfer is largely unknown. Here, the crystal structure of trimeric AP-B [(ApcD/ApcB)3] from Synechocystis sp. PCC 6803 at 1.75 Å resolution is reported. In the crystal lattice, eight trimers of AP-B form a porous, spherical, 48-subunit assembly of 193 Å in diameter with an internal cavity of 1.1 × 10(6) Å(3). While the overall structure of trimeric AP-B is similar to those reported for many other phycobiliprotein trimers, the chromophore pocket of the α-subunit, ApcD, has more bulky residues that tightly pack the phycocyanobilin (PCB). Ring D of the chromophores is further stabilized by close interactions with ApcB from the adjacent monomer. The combined contributions from both subunits render the conjugated rings B, C and D of the PCB in ApcD almost perfectly coplanar. Together with mutagenesis data, it is proposed that the enhanced planarity effectively extends the conjugation system of PCB and leads to the redshifted absorption (λmax = 669 nm) and fluorescence emission (679 nm) of the ApcD chromophore in AP-B, thereby enabling highly efficient energy transfer from the phycobilisome core to the reaction centres.


[Goniosynechiae as a complication of non-penetrating deep sclerectomy by inappropriate use of mydriatic: case report].
Non-penetrating deep sclerectomy has been an alternative to trabeculectomy, because it has the advantage of reducing intraocular pressure (IOP), without penetrating the anterior chamber. Thus, it avoids a sudden decompression and a plain anterior chamber. In this paper, we described a case of a female patient, 53 years old, black, with diagnosis of open-angle primary glaucoma, with unsatisfactory clinical control and who was submitted to non-penetrating deep sclerectomy. The patient developed goniosynechiae at the site of the sclerectomy, after inappropriate use of mydriatic eye-drops during the postoperative period, leading to an increase of IOP and, therefore, failure of the surgery. The use of mydriatic eye-drops during the postoperative period of deep sclerectomy is not indicated, because in this case, it led to surgery failure.


Protective role for proteoglycans against cationic lipid cytotoxicity allowing optimal transfection efficiency in vitro.
A dependence on proteoglycans for cationic lipid-mediated gene transfer has been suggested in previous studies [Mislick and Baldeschwieler (1996) Proc. Natl. Acad. Sci. U.S.A. 93, 12349-12354; Mounkes, Zhong, Cipres-Palacin, Heath and Debs (1998) J. Biol. Chem. 273, 26164-26170]. We have evaluated the mechanism of proteoglycan involvement in cationic lipid-mediated gene transfer. DNA plasmid uptake and gene expression were studied in wild-type Chinese hamster ovary (CHO) cells (CHO-K1), heparan sulphate-deficient CHO cells (pgsD-677) and proteoglycan-deficient CHO cells (pgsB-618). At an optimal ratio of cationic lipid to DNA, a substantial decrease in reporter gene expression was observed in proteoglycan-deficient cells compared with that in heparan sulphate-deficient and wild-type cells. However, there were no differences in reporter gene expression between the cell lines when transfected by electroporation. Moreover, all cell lines exhibited equal cationic-lipid-DNA complex uptake activities, as assessed by the measurement of intracellular (32)P-labelled and rhodamine-labelled DNA plasmid. An analysis of reflected-light images of wild-type and proteoglycan-deficient cells suggested that cationic lipids were preferentially toxic to proteoglycan-deficient cells. Cell-growth assays confirmed this, showing that cationic lipids exhibited a greater anti-proliferative activity in proteoglycan-deficient cells and in chlorate-treated wild-type cells than in the other cell lines. The growth-inhibitory effect of cationic lipids was abrogated by the addition of exogenous sulphated glycosaminoglycans. We conclude that the glycosaminoglycan part of proteoglycans serves a protective role against cationic lipid cytotoxicity, allowing optimal transfection efficiency in vitro.


Health-Related Quality of Life as a Prognostic Factor in Patients After Resection of Hepatic Malignancies.
Numerous studies have argued health-related quality of life (HRQoL) measures to be prognostic of survival in patients with chronic disease processes including cancer; however, only a few small studies have evaluated HRQoL changes in the setting of oncologic resections. The objectives of the present study were to investigate factors predicting HRQoL, the change in HRQoL over time, and HRQoL prognostic value in patients undergoing surgical resection of hepatic malignancies. We administered the Functional Assessment of Cancer Therapy-Hepatobiliary, Center for Epidemiologic Studies-Depression, Functional Assessment of Cancer Therapy-Fatigue, and Brief Pain Inventory to 128 patients with primary and metastatic hepatic malignancies enrolled between January 2008 to November 2011 and November 2013 to June 2015. Quality of life was obtained at the baseline, 4, 8, and 12 mo, using HRQoL questionnaires. The mean age of all patients included was 61 y, 42.6% had hepatocellular carcinoma, and 50.7% had metastatic colorectal carcinoma. HRQoL decreased from baseline at the 4-mo follow-up but stabilized to preoperative values at 8 and 12 mo. Depressive symptoms (P < 0.001), pain (P = 0.032), and fatigue (P < 0.001) were associated with HRQoL before surgery. Variables associated with HRQoL at 8 mo included extrahepatic recurrence (P = 0.002), depressive symptoms (P < 0.001), pain (P < 0.001), fatigue (P < 0.001), tumor macrovascular (P = 0.011), and microvascular invasion (P = 0.003). Using Cox regression and adjusting for demographics and disease-specific factors, preoperative HRQoL was significantly associated with overall survival. HRQoL is independently associated with survival in patients with liver malignancies undergoing surgical resection. Major curative liver surgery can be performed with short-term worsening of HRQoL but long-term improvement and stabilization in overall quality of life for patients with cancer.


Mean scalar concentration profile in a sheared and thermally stratified atmospheric surface layer.
Using only dimensional considerations, Monin and Obukhov proposed a "universal" stability correction function φ(c)(ζ) that accounts for distortions caused by thermal stratification to the mean scalar concentration profile in the atmospheric surface layer when the flow is stationary, planar homogeneous, fully turbulent, and lacking any subsidence. For nearly 6 decades, their analysis provided the basic framework for almost all operational models and data interpretation in the lower atmosphere. However, the canonical shape of φ(c)(ζ) and the departure from the Reynold's analogy continue to defy theoretical explanation. Here, the basic processes governing the scalar-velocity cospectrum, including buoyancy and the scaling laws describing the velocity and temperature spectra, are considered via a simplified cospectral budget. The solution to this cospectral budget is then used to derive φ(c)(ζ), thereby establishing a link between the energetics of turbulent velocity and scalar concentration fluctuations and the bulk flow describing the mean scalar concentration profile. The resulting theory explains all the canonical features of φ(c)(ζ), including the onset of power laws for various stability regimes and their concomitant exponents, as well as the causes of departure from Reynold's analogy.


[Cell therapy plus transmyocardial laser revascularization: a proposed alternative procedure for refractory angina].
We tested the hypothesis that TMLR combined with intramyocardial injection of BMC is safe, and may help increase the functional capacity of patient with refractory angina. Nine patients (eight men), 65+/-5 years old, with refractory angina for multivessel disease and previous myocardial revascularization procedures (CABG/PCI), not candidates for another procedure due to the extension of the disease were enrolled. TMLR (11+/-3 laser drills) was performed via a limited thoracotomy using a CO2 Heart Laser System. BMC were obtained immediately prior to surgery, and the lymphomonocytic fraction separated by density gradient centrifugation. During surgery, 5 mL containing approximately 1.9+/-0.3 x 10(8) BMC were delivered by multiple injections in the ischemic myocardium. Before (B) and 6 months (6M) after the procedure, patient underwent clinical evaluation and myocardial perfusion assessment by cardiac magnetic resonance imaging (MRI) during pharmacological stress with dypiridamole. No major complications or deaths occurred during the procedure. One patient died after 2 years (non cardiac cause). There was a reduction in the ischemic score as assessed by MRI from 1.64+/-0.10 (B) to 0.88+/-0.09 (6M) (P=0.01). Clinically, there was a reduction in functional class of angina from 3.7+/-0.2 (B) to 1.3+/-0.2 (6M) (P<0.0001). In this initial experience, the combined strategy of TMLR plus cell therapy appeared to be safe, and may have synergistically acted to reduce myocardial ischemia, with clinically relevant improvement in functional capacity. Provided these data are confirmed in a larger, randomized, controlled trial with longer follow-up, this strategy could be used as a novel therapeutic option for treating pt with refractory angina.


Bereaved informal cancer carers making sense of their palliative care experiences at home.
This qualitative study explored the positive meanings constructed and ascribed to the experience of providing palliative care at home by bereaved informal cancer carers, a group of individuals who are in a position to make sense of their caring experiences as a coherent whole. Twenty-two bereaved cancer carers, living in New South Wales (NSW), Australia, were interviewed as part of a larger mixed-method study examining the experience of informal cancer care. Participants were recruited through cancer support groups and cancer clinics, and through the Cancer Council NSW. Accounts of positive aspects of palliative caring were analysed using a thematic analytical approach from a constructionist perspective. The findings indicated that these bereaved carers gave accounts that accentuated the benefit and satisfaction derived from providing direct palliative care at home, which enabled them to construct positive meanings associated with their participation in the dying process, and as a result to ascribe subjectively meaningful interpretations to their loved ones' death and their sense of loss. This included a sense of reward for doing something good, meeting the expressed needs of the patient, continuing with normal life as much as possible, improving the conditions of the relationship and meeting cultural expectations of the right thing to do. Being present at the point of death was positioned as rewarding because it facilitated the process of saying goodbye, fostered inclusion of others, provided closure and was a spiritual experience. These findings suggest that there are positive and rewarding aspects associated with providing informal cancer care in a palliative context, and these aspects were pertinent and meaningful for carers in their endeavours to reconcile the difficulties and loss they experienced. This has implications for the prevention and amelioration of distress experienced by informal cancer carers, and suggests that future research should not ignore the positive aspects of providing palliative care.


The measurement of long-term health-related quality of life after injury: comparison of EQ-5D and the health utilities index.
Empirical head-to-head comparison of the health utility index (HUI) mark 2 and 3 and the EuroQol-5D (EQ-5D) in injury patients of all severity levels to obtain more insight into the strengths and limitations of the multi-attribute utility measures (MAUI) to estimate utility losses in injury populations. A self-assessment survey that included the EQ-5D, HUI2 and HUI3 to measure generic health-related quality of life. Injury patients in The Netherlands 2 years after they attended the emergency department. Shannon's index and Shannon's evenness index were used to assess absolute and relative informativity, both for the summary scores and by dimension. The study also analysed convergent and construct validity of the MAUI. Mean summary scores significantly differed between the instruments, with highest summary scores for HUI2 (0.88), followed by HUI3 (0.80) and EQ-5D (0.78). Absolute and relative informativity by dimension was highest for the HUI3 descriptive system. The HUI3 was most sensitive for ageing and comorbidity. The largest differences between the MAUI were found for pain/discomfort and anxiety/depression/emotion. The largest differences in discriminative power between EQ-5D and HUI (mark 2 and 3) were seen for skull-brain injury, internal organ injury and upper extremity fractures. Different MAUI resulted in significantly different summary scores. The instruments and their dimensions performed differently for injury severity levels, ageing, comorbidity and injury groups. A combination of the HUI and EQ-5D should be used in studies on injury-related disability, because the combination covers all relevant health dimensions, is applicable in all kinds of injury populations and in widely different age ranges.


Chemotactic factor inactivation by stimulated human neutrophils mediated by myeloperoxidase-catalyzed methionine oxidation.
Leukocyte chemoattractants were inactivated when exposed to human neutrophils and either ingestible particles or phorbol esters. Loss of biologic activity was time- and temperature-dependent, required physiologic concentrations of viable neutrophils and a halide, and was inhibited by azide or catalase. Neutrophils from patients with either hereditary myeloperoxidase deficiency or chronic granulomatous disease failed to inactivate the chemoattractants unless purified myeloperoxidase or H2O2, respectively, was added. Susceptibility to inactivation by neutrophils correlated with the presence of methionine in the attractant. Loss of chemotactic activity was blocked by low concentrations of methionine and by higher concentrations of other reducing agents, but was unaffected by oxidized methionine. Paper chromatography demonstrated that exposure of a formyl-methionyl peptide chemotactic factor to either the cellfree myeloperoxidase system or stimulated neutrophils resulted in its conversion to a molecular species whose location in the chromatographs was identical to that of the peptide containing oxidized methionine. Thus, stimulated human neutrophils inactivate peptide chemoattractants by secretion of myeloperoxidase and H2O2, which combine with halides to form oxidants that react with a critical methionine residue. We suggest that myeloperoxidase-catalyzed oxidation of thioethers may constitute an inflammatory control mechanism as well as a general means of modifying the functional properties of biologic mediators.


A Critical Review of State-of-the-Art and Emerging Approaches to Identify Fracking-Derived Gases and Associated Contaminants in Aquifers.
High-volume, hydraulic fracturing (HVHF) is widely applied for natural gas and oil production from shales, coals, or tight sandstone formations in the United States, Canada, and Australia, and is being widely considered by other countries with similar unconventional energy resources. Secure retention of fluids (natural gas, saline formation waters, oil, HVHF fluids) during and after well stimulation is important to prevent unintended environmental contamination, and release of greenhouse gases to the atmosphere. Here, we critically review state-of-the-art techniques and promising new approaches for identifying oil and gas production from unconventional reservoirs to resolve whether they are the source of fugitive methane and associated contaminants into shallow aquifers. We highlight future research needs and propose a phased program, from generic baseline to highly specific analyses, to inform HVHF and unconventional oil and gas production and impact assessment studies. These approaches may also be applied to broader subsurface exploration and development issues (e.g., groundwater resources), or new frontiers of low-carbon energy alternatives (e.g., subsurface H2 storage, nuclear waste isolation, geologic CO2 sequestration).


Surgical factors influence the outcome after Ivor-Lewis esophagectomy with intrathoracic anastomosis for adenocarcinoma of the esophagogastric junction: a consecutive series of 240 patients at an experienced center.
Despite a considerable number of randomized studies, the surgical approach to locally advanced adenocarcinoma of the esophagogastric junction (AEG) I and II is still discussed controversially. Thus, we evaluated the surgical risk and outcome after an abdominothoracic esophagectomy (Ivor-Lewis) with intrathoracic anastomosis as standard procedure. Between 1998 and 2006, a total of 240 consecutive patients underwent standardized right thoracoabdominal esophagectomy with two-field lymphadenectomy and intrathoracic anastomosis (Ivor-Lewis operation) for AEG I (n = 206) or AEG II (n = 34). A total of 157 patients (65.4%) had neoadjuvant chemotherapy. Postoperative morbidity occurred in 17.9% (43 of 240). Overall mortality was 3.8% (9 of 240). The majority of patients (4 of 9) died because of severe pulmonary complications (44.4%) irrespective of surgical complications. Neoadjuvant chemotherapy did not increase morbidity or mortality. The median overall survival was 51 months. Multivariate analysis including age >75 years, clinical response to chemotherapy, complications, R-category and N-category revealed R-category (P = .005; relative risk [RR] 0.32, 95% confidence interval [95% CI] 0.14-0.70) and complications (P < .001, RR 0.16, 95% CI 0.08-0.35) as independent prognostic factors for all patients. Complications was the only independent prognostic factor (P < .001, RR 0.09, 95% CI 0.08-0.35) for the R0 resected patients. At an experienced center, Ivor-Lewis resection is a safe surgical procedure. Outcome of patients was significantly influenced by surgical factors such as complete resection and complications. Neoadjuvant chemotherapy did not lead to higher morbidity and mortality. The high mortality from non-surgery-related complications emphasizes the importance of careful preoperative evaluation of comorbidities and patient selection.


Social and personal consequences of disability in adults with hip and knee arthroplasty. A French national community based survey.
To describe and compare participation restrictions and environmental factors of persons with and without hip or knee arthroplasty in a national community based survey: the Handicap, Disability, Dependence Survey. During the 1999 French Census, a screening questionnaire was proposed to 417,500 persons, for which the response rate was 86%. A stratified random sample with an overrepresentation of disabled persons was performed to constitute the selected population. A computer assisted interview was proposed to 21,760 persons, with a 78% response rate. Chronic conditions, impairment, disability, participation restrictions, and the description of environmental factors were ascertained from the subjects' reports. The hip and knee arthroplasty group was estimated at 691,000 persons in the French population. Although reporting a higher level of disability, this population did not report more participation restrictions than the general population in terms of their economic situation, housing, social relationships, and holidays. Moreover, when comparing environmental factors, this population reported better housing accessibility, more assistive devices (OR 5.2, 95% CI 3.7-7.2), specific fittings (OR 2.9, 95% CI 2.0-4.2), and helpers (OR 1.8, 95% CI 1.3-2.5). These environmental factors may have compensated for the higher level of disability. Nevertheless, individuals with hip and knee arthroplasty reported more disadvantages when moving within their environment (OR 2.1, 95% CI 1.5-2.9). This study provides a detailed description based on a national random sample of participation restrictions and environmental factors of adults with hip and knee arthroplasty.


Computed tomography angiography versus angiography for guiding percutaneous coronary interventions in bifurcation lesions - A prospective randomized pilot study.
There is no data on the impact of coronary computed tomography angiography (coronary CTA), as an addition to angiography, on the outcomes of percutaneous coronary interventions (PCI) in bifurcation lesions. Patients with stable coronary artery disease scheduled for elective bifurcation PCI were randomized 1:1 to planning the procedure based on coronary CTA and angiography (CTA group) or angiography alone (CA group). The primary efficacy endpoint was the immediate angiographic result. Secondary efficacy endpoints were: a) procedural characteristics and b) postprocedural fractional flow reserve (FFR) in the side branch (SB) in a subgroup of patients. Safety outcomes were: a) periprocedural myocardial infarction, b) contrast use and c) radiation dose. PCI of 45 lesions in the CTA group and 47 lesions in the CA group was performed. Postprocedural lumen diameters in the main branch (MB) and SB, frequency of SB compromise or occlusion and mean SB FFR values were not different between study groups. Two or more stents were implanted less frequently in the CTA group than in the CA group (18% vs. 43%, p = 0.01). This difference was driven by less frequent MB 2-stent overlap in the CTA group (7 vs. 21%, p = 0.046) and numerically less SB stenting (11% vs. 21%, p = 0.07). Proximal optimization technique was used more frequently in the CTA group (44% vs. 21%, p = 0.018). CTA-assisted bifurcation PCI leads to similar immediate results compared with angiography alone, however is associated with higher use of single-stent procedures with proximal optimization, less frequent 2-stent overlap and less SB stenting.


Drying banana seeds for ex situ conservation.
The ability of seeds to withstand drying is fundamental to ex situ seed conservation but drying responses are not well known for most wild species including crop wild relatives. We look at drying responses of seeds of Musa acuminata and Musa balbisiana, the two primary wild relatives of bananas and plantains, using the following four experimental approaches: (i) We equilibrated seeds to a range of relative humidity (RH) levels using non-saturated lithium chloride solutions and subsequently measured moisture content (MC) and viability. At each humidity level we tested viability using embryo rescue (ER), tetrazolium chloride staining and germination in an incubator. We found that seed viability was not reduced when seeds were dried to 4% equilibrium relative humidity (eRH; equating to 2.5% MC). (ii) We assessed viability of mature and less mature seeds using ER and germination in the soil and tested responses to drying. Findings showed that seeds must be fully mature to germinate and immature seeds had negligible viability. (iii) We dried seeds extracted from ripe/unripe fruit to 35-40% eRH at different rates and tested viability with germination tests in the soil. Seeds from unripe fruit lost viability when dried and especially when dried faster; seeds from ripe fruit only lost viability when fast dried. (iv) Finally, we dried and re-imbibed mature and less mature seeds and measured embryo shrinkage and volume change using X-ray computer tomography. Embryos of less mature seeds shrank significantly when dried to 15% eRH from 0.468 to 0.262 mm3, but embryos of mature seeds did not. Based on our results, mature seeds from ripe fruit are desiccation tolerant to moisture levels required for seed genebanking but embryos from immature seeds are mechanistically less able to withstand desiccation, especially when water potential gradients are high.


Potassium exchange in the whole cell, cytoplasm, and nucleus of amphibian oocytes.
Potassium isotope exchange was studied in whole oocytes, with and without ovarian follicles, and in oocyte cytoplasm and nucleus. Cryomicrodissection was used to prevent solute redistribution during nuclear and cytoplasmic separation. Manual follicle removal causes a small decrease in the K+ of the preparation. No effect of follicle removal is seen on 42K+ exchange. Whole oocyte exchange is multiphasic and reflects the presence of two intracellular K+ fractions. One of these fractions is present in both nucleus and cytoplasm. It exhibits first-order exponential kinetics, apparently established at the cell membrane. The second fraction is restricted to cytoplasm and exchanges at an imperceptible rate. The fractions differ in pre- and posthibernation oocytes. These observations clarify the mechanism whereby nuclear/cytoplasmic K+ concentration asymmetries are maintained and the mechanisms responsible for the high K+-activity coefficient previously reported in these cells.


Immunomodulatory activities of alpha-mangostin on peripheral blood mononuclear cells.
Mangosteen (Garcinia mangostana L.) a tropical fruit, has been used in traditional medicine. A frequently used part of mangosteen is the pericarp, containing a high content of xanthones. alpha-Mangostin, one of the major xanthone derivatives, exhibits a variety of actions, including antimicrobial, antioxidant, cytotoxic and antitumor; however, its function on the immune system is still equivocal. This study aimed to examine the immunomodulatory activities of alpha-mangostin on lymphocyte lineage and cytokine production in human peripheral blood mononuclear cells (PBMCs). The cytotoxic activity of alpha-mangostin was measured by MTT assay. The concentration of alpha-mangostin at 5.55 microg/mL resulted in a 50% survival of PBMCs, which was as potent a cytotoxic activity as that of paclitaxel. After 24 h of PBMCs culture, the percentages of T cells (CD3+), B cells (CD19+) and NK cells (CD3-CD16+CD56+) were not significantly changed by treatment with 1, 2 and 4 microg/mL of alpha-mangostin compared with untreated-PBMCs; in addition, the percentages of these lymphocytes treated with the combination of alpha-mangostin (1, 2 and 4 microg/mL) and the mitogen concanavalin A (ConA) was not significantly different from that of ConA-treated PBMCs. For cytokine secretion, alpha-mangostin (1, 2 and 4 microg/mL) did not significantly induce either proinflammatory cytokines (i.e., TNF-alpha and IL-1beta) or cytokine of adaptive immunity (i.e., IL-2). The combination of alpha-mangostin (1, 2 and 4 microg/mL) and ConA did not significantly alter the relative difference of TNF-alpha and IL-1beta compared with ConA-treated PBMCs; however, these combinations could significantly decrease the relative difference of IL-2 compared with ConA-treated PBMCs. These data indicated that alpha-mangostin was able to inhibit IL-2 release without interfering with human immune cells; therefore, further studies are necessary to investigate its effect on IL-2 production.


Functional constipation as a neglected condition in laryngectomized patients.
When considering the quality of life of patients who undergo total laryngectomy, constipation problems are often overlooked. This study aims to determine whether surgical or concurrent chemoradiotherapy treatments matter in the development of functional constipation in patients with advanced-stage laryngeal cancer. Patients without presurgical constipation (according to Rome IV) who were treated with either surgery (n=30) or chemoradiation (n=25) were evaluated for constipation development. Interim evaluation and recommendations were made on the 3rd month. On the 3rd and 6th months, the frequency of constipation between groups was compared using the Rome IV criteria and European Organization for Research and Treatment of Cancer Quality of Life Scale-C30 constipation symptom score. On the 3rd month, the frequency of constipation was higher in the surgery group than in the chemoradiation group (76.7% vs. 28%, p<0.001). On the 6th month, although still higher, the frequency of constipation decreased following the recommendations in laryngectomized patients (56.7% vs. 36%, p=0.17). C30 Constipation "yes/no" inquiry failed to detect 11 (36.6%) and 8 (26.6%) laryngectomized patients with constipation on the 3rd and 6th months, respectively. Constipation is more frequent and develops in the early period of postlaryngectomy. Patients seem to benefit from additional special rehabilitation recommendations. Constipation-prone laryngectomized patients should be screened with the Rome IV criteria, which effectively detect constipation.


Initiation of Flower Buds in Rhododendron after Application of Growth Retardants.
Vegetative terminals of azalea (Rhododendron spp.) plants of varying ages initiated flower buds promptly after application of the chemical growth retardants, tributyl-2,4-dichlorobenzylphosphonium chloride (phosfon) and (2-chloroethyl) trimethylammonium chloride (CCC), as soil drenches. This response occurred under environmental conditions which prevented or limited flower bud initiation in untreated plants. Normal flowering followed exposure of the treated plants to dormancy-breaking cool storage.


Effect of stretching the superior vena cava on heart rate in rats.
The effect on the heart rate of stretching the superior vena cava 3-4 mm above its entrance to the right atrium was examined in rats anesthetized with chloralose. The superior vena cava was stretched 41 times in 16 rats. There was a mean increase of 24.1 +/- 1.9 (SE) beats/min. This response was abolished (6 rats) by sympathetic blockade (with propranolol hydrochloride and bretylium tosylate) and by bilateral vagosympathectomy (5 rats). Application of lidocaine to the cervical vagi also abolished the response in a reversible manner (4 rats). In five rats, histological examination of the superior vena cava revealed an abundance of nerves, which formed a net in the stretched region. Complex unencapsulated nerve endings similar to those demonstrated in the dog and cat were not found.


Closer look at time averages of the logistic map at the edge of chaos.
The probability distribution of sums of iterates of the logistic map at the edge of chaos has been recently shown [U. Tirnakli, Phys. Rev. E 75, 040106(R) (2007)] to be numerically consistent with a q-Gaussian, the distribution which-under appropriate constraints-maximizes the nonadditive entropy Sq, which is the basis of nonextensive statistical mechanics. This analysis was based on a study of the tails of the distribution. We now check the entire distribution, in particular, its central part. This is important in view of a recent q generalization of the central limit theorem, which states that for certain classes of strongly correlated random variables the rescaled sum approaches a q-Gaussian limit distribution. We numerically investigate for the logistic map with a parameter in a small vicinity of the critical point under which conditions there is convergence to a q-Gaussian both in the central region and in the tail region and find a scaling law involving the Feigenbaum constant delta. Our results are consistent with a large number of already available analytical and numerical evidences that the edge of chaos is well described in terms of the entropy Sq and its associated concepts.


Missing metal-linker connectivities in a 3-D robust sulfonate-based metal-organic framework for enhanced proton conductivity.
A rare example of a robust, sulfonate-based metal-organic framework (MOF) with a prototypical primitive-cubic topology has been successfully synthesized. Importantly, one-fourth of the metal-ligand connectivities residing in the MOF are missing, resulting in the appearance of coordinatively accessible Cu2+ sites and pendant sulfonate groups. This unprecedented crystal irregularity phenomenon of MOFs substantially enhances their affinity towards water molecules, thus increasing the proton conductivity by two orders of magnitude.


Beef flavor: a review from chemistry to consumer.
This paper briefly reviews research that describes the sensation, generation and consumer acceptance of beef flavor. Humans sense the five basic tastes in their taste buds, and receptors in the nasal and sinus cavities sense aromas. Additionally, trigeminal senses such as metallic and astringent are sensed in the oral and nasal cavities and can have an effect on the flavor of beef. Flavors are generated from a complex interaction of tastes, tactile senses and aromas taken collectively throughout the tongue, nasal, sinus and oral cavities. Cooking beef generates compounds that contribute to these senses and result in beef flavor, and the factors that are involved in the cookery process determine the amount and type of these compounds and therefore the flavor generated. A low-heat, slow cooking method generates primarily lipid degradation products, while high-heat, fast cookery generates more Maillard reaction products. The science of consumer acceptance, cluster analyses and drawing relationships among all flavor determinants is a relatively new discipline in beef flavor. Consumers rate beef that has lipid degradation products generated from a low degree of doneness and Maillard flavor products from fast, hot cookery the highest in overall liking, and current research has shown that strong relationships exist between beef flavor and consumer acceptability, even more so than juiciness or tenderness.


Cytogenetics and molecular genetics of malignant gliomas and medulloblastoma.
Malignant gliomas and medulloblastomas which are the most common primary malignant brain tumours of adult and children, respectively, resemble other neurogenic tumours as they frequently contain gene amplification and show non-random loss of specific chromosomal regions. In gliomas the gene which is most often amplified, is the epidermal growth factor receptor gene. In many cases the gene is rearranged as well, producing abnormally small epidermal growth factor receptor proteins. More than 80% of tumours have lost chromosome 10 and losses of 9p13, 17p and 22 occur in subgroups of cases. 17p loss is associated with point mutations of the p53 gene, but the relevant genes in the other chromosomal regions remain to be identified. For medulloblastoma the most frequent chromosomal abnormality is i(17q). Whether or not p53 gene mutations are the targets of 17p losses in these tumours remains to be determined. Approximately 5% of medulloblastoma biopsies contain gene amplification, although the incidence in medulloblastoma cell lines is more than 80%. c-myc is the gene which is most frequently amplified in this tumour type. The relationship of these various molecular genetic abnormalities to the biology of the tumours and the course of the patients remains largely unexplored.


[Justice and equity as determinants of medical decision-making].
Every medical decision-making is to do justice to the one who suffers and who considers his pain as nonsensical. This requirement falls into 3 universal ethical principles, governing the patient-doctor relationship: the autonomy of the person even if it is a young child, beneficence and non-maleficence. Adhering to these principles gives medical decision-making its ethical dimension. It implies that the doctor makes the best use of the emotions that he feels in front of his patient and/or in front of his relatives: respect for autonomy, the beneficent compassion and the fear of maleficence in diagnosis and care. If a paediatrician combines these 3 affects harmoniously, his attitude is in keeping with the requirement of justice, which is the quintessence of Ethics. However, reality is often more complex, a source of conflicting emotions and, in fine, a source of an anguish whose benefit is yet to alert on the necessity to carry on looking for the right decision-making: the emotional revision consists of a meta-analysis of the objective and subjective data of the problem. It preciously helps to establish a compromise of justice. Eventually, in order to be just, the doctor must include the concern of equity amongst the criteria of his decision-making, in other words a fair allocation of the goods and of the care services. Unfortunately, this concern is undermined by geopolitical, socio-economic and cultural factors, which vary greatly according to the environmental conditions that might mar what should be optimal ethical decisionmaking. The doctor cannot solve these problems on his own, but he has to know them in order to deal with them.


Differential regulation of nitric oxide synthase isoforms in experimental acute chagasic cardiomyopathy.
We have previously demonstrated induction and high level expression of IL-1beta, IL-6 and tumour necrosis factor-alpha in the myocardium during the acute stage of experimental Trypanosoma cruzi infection (Chagas' disease). The myocardial depressive effects of these cytokines are mediated in part by the induction of nitric oxide synthase (NOS), production of nitric oxide (NO) and formation of peroxynitrite. In this study we investigated the expression, activity and localization of NOS isoforms, and the levels of NO, malondialdehyde (a measure of oxidative stress), and peroxynitrite in rats at 1.5, 5, 10 and 15 days after infection with T. cruzi trypomastigotes. The myocardial inflammatory infiltrate and number of amastigote nests increased over the course of infection. A significant increase in tissue nitrate + nitrite levels, NOS2 mRNA, and NOS2 enzyme activity was observed at all time points in the infected compared with uninfected animals. The enzyme activity of constitutive NOS, tissue malondialdehyde levels, and NOS3 mRNA levels was only transiently increased after infection. The protein levels of the NOS isoforms paralleled their mRNA expression. While no positive nitrotyrosine immunoreactivity was detected in control myocardium, its levels increased in infected animals over time. Thus, by 1.5 days post-infection, when no parasite or immune cell infiltration could be detected, the myocardium expressed high levels of NOS and NO metabolites. Nevertheless, the early production of NO in the myocardium was not sufficient to clear the parasites.


Application of culturomics in fungal isolation from mangrove sediments.
Fungi play a crucial role in ecosystems, and they have been widely considered a promising source for natural compounds that are crucial for drug discovery. Fungi have a high diversity, but about 95% of them remain unknown to science. The description rate of fungi is very low, mainly due to the inability of most fungi to grow in artificial media, which could not provide a sufficiently similar environment to their natural habitats. Moreover, many species in nature are in a state of low metabolic activity which cannot readily proliferate without proper resuscitation. Previously developed culturomics techniques are mostly designed and applicable for bacteria, with few attempts for fungal isolation because of their significantly larger cell size and hyphal growth properties. This study attempted to isolate previously uncultured and rare fungi from mangrove sediments using newly developed fungal enrichment culture method (FECM) and fungal isolation chips (FiChips). Comparison of fungal community composition at different enrichment stages showed that FECM had great influence on fungal community composition, with rare taxa increased significantly, thus improving the isolation efficiency of previously uncultured fungi. Similarly, in situ cultivation using FiChips has a significant advantage in detecting and culturing rare fungi, as compared to the conventional dilution plate method (DPM). In addition, based on morphological comparisons and phylogenetic analyses, we described and proposed 38 new ascomycetous taxa, including three new families, eight new genera, 25 new species, and two new combinations (presented in additional file 1). Our study demonstrated that mangrove sediments harbor a high diversity of fungi, and our new isolation approaches (FECM and FiChips) presented a high efficiency in isolating hitherto uncultured fungi, which is potentially usable for fungal isolation in other similar environments. Video Abstract.


Examination of the clonal deletion hypothesis following transfusions in rat cardiac transplants.
To test the clonal deletion hypothesis, different levels of immunization were carried out by 0, 1, 2, or 3 transfusions preoperatively, together with 10 methods of immunosuppression. In this way, it was hoped that we could determine the optimal way to immunize and the optimal immunosuppressant treatment to deactivate the responding cells. With the Buffalo-to-Lewis rat heart allograft model, the control mean survival rate was 7 days. Rats with 2 or 3 transfusions had 5- and 12-day survival rates, respectively. If azathioprine was given in addition before grafting, 19- and 18-day survival rates were seen, and if azathioprine was given after grafting, extended survivals of 33 and 34 days were achieved. The longest survival rate of greater than 52 days was obtained by a single transfusion and 25 mg/kg/day of cyclophosphamide given before transplantation. Splenectomy following 2 or 3 transfusions prolonged survival rates to 16-18 days, suggesting that mechanical removal of immunized cells is somewhat effective. Most of the antibody produced by one transfusion was IgM, as were antibodies that resulted from transfusions followed by azathioprine. It is possible that reduction of IgG-producing cells is important. These results are consistent with the clonal deletion theory, although they do not necessarily provide final proof. The experiments suggest that the optimal transfusion effect may be obtained with minimal immunization (1 transfusion) and proper immunosuppression before transplantation.


Guest-induced pore breathing controls the spin state in a cyanido-bridged framework.
Iron(ii) spin cross-over (SCO) compounds combine a thermally driven transition from the diamagnetic low-spin (LS) state to the paramagnetic high-spin (HS) state with a distinct change in the crystal lattice volume. Inversely, if the crystal lattice volume was modulated post-synthetically, the spin state of the compound could be tunable, resulting in the inverse effect for SCO. Herein, we demonstrate such a spin-state tuning in a breathing cyanido-bridged porous coordination polymer (PCP), where the volume change resulting from guest-induced gate-opening and -closing directly affects its spin state. We report the synthesis of a three-dimensional coordination framework {[FeII(4-CNpy)4]2[WIV(CN)8]·4H2O}n (1·4H2O; 4-CNpy = 4-cyanopyridine), which demonstrates a SCO phenomenon characterized by strong elastic frustration. This leads to a 48 K wide hysteresis loop above 140 K, but below this temperature results in a very gradual and incomplete SCO transition. 1·4H2O was activated under mild conditions, producing the nonporous {[FeII(4-CNpy)4]2[WIV(CN)8]}n (1) via a single-crystal-to-single-crystal process involving a 7.3% volume decrease, which shows complete and nonhysteretic SCO at T1/2 = 93 K. The low-temperature photoswitching behavior in 1 and 1·4H2O manifested the characteristic elasticity of the frameworks; 1 can be quantitatively converted into a metastable HS state after 638 nm light irradiation, while the photoactivation of 1·4H2O is only partial. Furthermore, nonporous 1 adsorbed CO2 molecules in a gated process, leading to {[FeII(4-CNpy)4]2[WIV(CN)8]·4CO2}n (1·4CO2), which resulted in a 15% volume increase and stabilization of the HS state in the whole temperature range down to 2 K. The demonstrated post-synthetic guest-exchange employing common gases is an efficient approach for tuning the spin state in breathing SCO-PCPs.


Exon skipping and gene transfer restore dystrophin expression in hiPSC-cardiomyocytes harbouring DMD mutations.
With an incidence of ~1:3,500 to 5,000 in male children, Duchenne muscular dystrophy (DMD) is an X-linked disorder in which progressive muscle degeneration occurs and affected boys usually die in their twenties or thirties. Cardiac involvement occurs in 90% of patients and heart failure accounts for up to 40% of deaths. To enable new therapeutics such as gene therapy and exon skipping to be tested in human cardiomyocytes, we produced human induced pluripotent stem cells (hiPSC) from seven patients harbouring mutations across the DMD gene. Mutations were retained during differentiation and analysis indicated the cardiomyocytes showed a dystrophic gene expression profile. Antisense oligonucleotide-mediated skipping of exon 51 restored dystrophin expression to ~30% of normal levels in hiPSC-cardiomyocytes carrying exon 47-50 or 48-50 deletions. Alternatively, delivery of a dystrophin minigene to cardiomyocytes with a deletion in exon 35 or a point mutation in exon 70 allowed expression levels similar to those seen in healthy cells. This demonstrates that DMD hiPSC-cardiomyocytes provide a novel tool to evaluate whether new therapeutics can restore dystrophin expression in the heart.


Convergence between temperament ratings in early infancy.
This study assesses the degree of convergence between two temperament questionnaires promoted for use with relatively young infants. The mothers of 90 normal, healthy infants were asked to complete the infant Behavior Questionnaire (IBQ) and the Early Infancy Temperament Questionnaire (EITQ) when their babies were approximately 3 months of age; the returned questionnaires were reduced into the dimensions defined by the instruments' authors. The EITQ includes a General Impressions rating scale that provides a more global estimate of the same temperament dimensions, and these scores were also examined. Infant ratings on both questionnaires were similar in magnitude to results of other published reports. Comparisons across instruments showed that the IBQ and EITQ seem to measure similar aspects of behavioral style, because all of the IBQ dimensions were significantly related to the appropriate EITQ dimensions. These results support the validity of both questionnaires for use with infants in the first months of life.


The gp116 of the gp58/116 complex of human cytomegalovirus represents the amino-terminal part of the precursor molecule and contains a neutralizing epitope.
The glycoprotein complex gp58/116 of human cytomegalovirus (HCMV) represents a dominant antigen for the humoral immune response. We have used the human monoclonal antibody C23, which is capable of neutralizing HCMV in tissue culture without the addition of complement, to study the origin of gp116 as well as the amino acid sequence recognized by the antibody. Our results show that gp116 is derived from the same open reading frame as gp58 and that it represents the amino-terminal portion of the precursor protein. Using prokaryote-expressed beta-galactosidase-gp116 fusion proteins, the binding site of C23 was located to between amino acids 27 to 84 of the amino-terminal portion of gp116. Analyses of HCMV-positive human sera revealed that this portion of the molecule is immunogenic during natural infection.


Building a bright, evidence-informed future: a conversation starter from the incoming editors.
Health Research and Policy Systems (HARPS) has gone from strength to strength since it was established in 2003. As new Editors-in-Chief, we look forward to a bright future for HARPS, and we would like to start a conversation with you, HARPS readers, authors, editors and others, about how HARPS can best support ongoing progress and debate on evidence-informed health research policy and systems, particularly in developing countries. As a starting point for discussion, we would like to highlight three areas that we are passionate about, namely supporting an integrated community of researchers and policy-makers; building a focus on how health research and policy systems can support achievement of the Sustainable Development Goals; and strengthening our commitment to communicating and disseminating the work published in HARPS. We invite you to contribute your thoughts, ideas and suggestions on the future of HARPS, as we work together towards an evidence-informed future.


[Mutants of Bacillus subtilis--producer of alkaline proteinase, sporulating in the presence of high concentrations of glucose].
Glur mutants of Bacillus subtilis A-50 capable of sporulating in the medium containing 5% glucose are isolated. The mutants are divided into three groups. The first group comprises glur mutants whose level of proteolytic activity is lower than that of the initial strain A-50. The productivity of mutants of the 2nd group is equal to the level of alkaline proteinase synthesis characteristics of the original strain. In mutants of the third group the level of alkaline protease synthesis exceeds the productivity of the initial strain. There was no correlation between the efficiency of spore formation in the mutants obtained and the level of alkaline protease activity. The mutants of different groups differ in their ability to grow on different sugars.


A phosphatidylserine species inhibits a range of TLR- but not IL-1beta-induced inflammatory responses by disruption of membrane microdomains.
TLRs detect conserved molecular patterns that are unique to microbes, enabling tailored responses to invading pathogens and modulating a multitude of immunopathological conditions. We investigated the ability of a naturally occurring stearoyl-arachidonoyl form of phosphatidylserine (SAPS) to inhibit the proinflammatory effects of TLR agonists in models of inflammation investigating the interaction of leukocytes with epithelial and endothelial cells. The responses to LPS of both epithelial and endothelial cells were highly amplified in the presence of PBMCs. Coincubation with SAPS markedly inhibited activation of cocultures by LPS, principally through inhibition of the TLR4 signaling pathway in PBMCs; however, this was not through downmodulation of TLR4 or coreceptor expression, nor was IL-1beta-induced cytokine release affected. SAPS also impaired Pam(3)CSK(4) (TLR2/1), Gardiquimod (TLR7/8), and Streptococcus pneumoniae-induced cytokine release, but had only modest effects on poly(I:C) (TLR3)-induced responses. Fluorescence resonance energy transfer analysis of molecular associations revealed that SAPS disrupted the association of both TLR4 and TLR2 with their respective membrane partners that are required for signaling. Thus, our data reinforce the existence and importance of cooperative networks of TLRs, tissue cells, and leukocytes in mediating innate immunity, and identify a novel disrupter of membrane microdomains, revealing the dependence of TLR signaling on localization within these domains.


Novel ethanol production using biomass preprocessing to increase ethanol yield and reduce overall costs.
Ethanol biorefineries need to lower their overall production costs to become economically feasible. Two strategies to achieve this are to reduce costs using cheaper feedstocks or to increase the ethanol production yield. Low-cost feedstocks usually have high non-structural components (NSC) content; therefore, a new process is necessary to accommodate these feedstocks and overcome the negative effects of NSC. This study developed a novel ethanol biorefinery process including a biomass preprocessing step that enabled the use of lower-cost feedstocks while improving ethanol production without detoxification (overliming). Two types of poplar feedstocks were used, low-quality whole-tree chips (WTC) and high-quality clean pulp chips (CPC), to determine if the proposed process is effective while using feedstocks with different NSC contents. Technical assessment showed that acidic preprocessing increased the monomeric sugar recovery of WTC from 73.2% (untreated) to 87.5% due to reduced buffering capacity of poplar, improved sugar solubilization during pretreatment, and better enzymatic hydrolysis conversion. Preprocessing alone significantly improved the fermentability of the liquid fraction from 1-2% to 49-56% for both feedstocks while overliming improved it to 45%. Consequently, it was proposed that preprocessing can substitute for the detoxification step. The economic assessment revealed that using poplar WTC via the new process increased annual ethanol production of 10.5 million liters when compared to using CPC via overliming (base case scenario). Also, savings in total operating costs were about $10 million per year when using cheaper poplar WTC instead of CPC, and using recycled water for preprocessing lowered its total operating costs by 45-fold. The novel process developed in this study was successful in increasing ethanol production while decreasing overall costs, thus facilitating the feasibility of lignocellulosic ethanol biorefineries. Key factors to achieving this outcome included substituting overliming by preprocessing, enabling the use of lower-quality feedstock, increasing monomeric sugar recovery and ethanol fermentation yield, and using recycled water for preprocessing. In addition, preprocessing enabled the implementation of an evaporator-combustor downstream design, resulting in a low-loading waste stream that can be treated in a wastewater treatment plant with a simple configuration.


Cannabinoid Regulation of Fear and Anxiety: an Update.
Anxiety- and trauma-related disorders are prevalent and debilitating mental illnesses associated with a significant socioeconomic burden. Current treatment approaches often have inadequate therapeutic responses, leading to symptom relapse. Here we review recent preclinical and clinical findings on the potential of cannabinoids as novel therapeutics for regulating fear and anxiety. Evidence from preclinical studies has shown that the non-psychotropic phytocannabinoid cannabidiol and the endocannabinoid anandamide have acute anxiolytic effects and also regulate learned fear by dampening its expression, enhancing its extinction and disrupting its reconsolidation. The findings from the relevant clinical literature are still very preliminary but are nonetheless encouraging. Based on this preclinical evidence, larger-scale placebo-controlled clinical studies are warranted to investigate the effects of cannabidiol in particular as an adjunct to psychological therapy or medication to determine its potential utility for treating anxiety-related disorders in the future.


HilE represses the activity of the Salmonella virulence regulator HilD via a mechanism distinct from that of intestinal long-chain fatty acids.
The expression of virulence factors essential for the invasion of host cells by Salmonella enterica is tightly controlled by a network of transcription regulators. The AraC/XylS transcription factor HilD is the main integration point of environmental signals into this regulatory network, with many factors affecting HilD activity. Long-chain fatty acids, which are highly abundant throughout the host intestine, directly bind to and repress HilD, acting as environmental cues to coordinate virulence gene expression. The regulatory protein HilE also negatively regulates HilD activity, through a protein-protein interaction. Both of these regulators inhibit HilD dimerization, preventing HilD from binding to target DNA. We investigated the structural basis of these mechanisms of HilD repression. Long-chain fatty acids bind to a conserved pocket in HilD, in a comparable manner to that reported for other AraC/XylS regulators, whereas HilE forms a stable heterodimer with HilD by binding to the HilD dimerization interface. Our results highlight two distinct, mutually exclusive mechanisms by which HilD activity is repressed, which could be exploited for the development of new antivirulence leads.


Effects of Six Sequential Charged Particle Beams on Behavioral and Cognitive Performance in B6D2F1 Female and Male Mice.
The radiation environment astronauts are exposed to in deep space includes galactic cosmic radiation (GCR) with different proportions of all naturally occurring ions. To assist NASA with assessment of risk to the brain following exposure to a mixture of ions broadly representative of the GCR, we assessed the behavioral and cognitive performance of female and male C57BL/6J × DBA2/J F1 (B6D2F1) mice two months following rapidly delivered, sequential 6 beam irradiation with protons (1 GeV, LET = 0.24 keV, 50%), 4He ions (250 MeV/n, LET = 1.6 keV/μm, 20%), 16O ions (250 MeV/n, LET = 25 keV/μm 7.5%), 28Si ions (263 MeV/n, LET = 78 keV/μm, 7.5%), 48Ti ions (1 GeV/n, LET = 107 keV/μm, 7.5%), and 56Fe ions (1 GeV/n, LET = 151 keV/μm, 7.5%) at 0, 25, 50, or 200 cGy) at 4-6 months of age. When the activity over 3 days of open field habituation was analyzed in female mice, those irradiated with 50 cGy moved less and spent less time in the center than sham-irradiated mice. Sham-irradiated female mice and those irradiated with 25 cGy showed object recognition. However, female mice exposed to 50 or 200 cGy did not show object recognition. When fear memory was assessed in passive avoidance tests, sham-irradiated mice and mice irradiated with 25 cGy showed memory retention while mice exposed to 50 or 200 cGy did not. The effects of radiation passive avoidance memory retention were not sex-dependent. There was no effect of radiation on depressive-like behavior in the forced swim test. There was a trend toward an effect of radiation on BDNF levels in the cortex of males, but not for females, with higher levels in male mice irradiated with 50 cGy than sham-irradiated. Finally, sequential 6-ion irradiation impacted the composition of the gut microbiome in a sex-dependent fashion. Taxa were uncovered whose relative abundance in the gut was associated with the radiation dose received. Thus, exposure to sequential six-beam irradiation significantly affects behavioral and cognitive performance and the gut microbiome.


Inhibiting Phase Transfer of Protein Nanoparticles by Surface Camouflage-A Versatile and Efficient Protein Encapsulation Strategy.
Engineering a system with a high mass fraction of active ingredients, especially water-soluble proteins, is still an ongoing challenge. In this work, we developed a versatile surface camouflage strategy that can engineer systems with an ultrahigh mass fraction of proteins. By formulating protein molecules into nanoparticles, the demand of molecular modification was transformed into a surface camouflage of protein nanoparticles. Thanks to electrostatic attractions and van der Waals interactions, we camouflaged the surface of protein nanoparticles through the adsorption of carrier materials. The adsorption of carrier materials successfully inhibited the phase transfer of insulin, albumin, β-lactoglobulin, and ovalbumin nanoparticles. As a result, the obtained microcomposites featured with a record of protein encapsulation efficiencies near 100% and a record of protein mass fraction of 77%. After the encapsulation in microcomposites, the insulin revealed a hypoglycemic effect for at least 14 d with one single injection, while that of insulin solution was only ∼4 h.


Ten maxims for out of class learning to outclass the academic challenges of COVID-19.
This article was migrated. The article was marked as recommended. Responding to the COVID-19 pandemic, universities world over are aggressively moving their learning, teaching and assessment to online platforms. Educators suddenly feel unsettled, unprepared and unsupported. This inevitable segue from a traditional to online environment will need faculty to grapple quickly and efficiently with the technological imperatives. The purpose of this article is to present ten maxims to navigate the academic challenges posed by out of class learning. The article includes a user-friendly checklist to facilitate the blooming of online teachers.


Carbon nanotubes for transdermal drug delivery.
In this study, drug carrier properties through skin and penetration enhancement effects of carbon nanotubes (CNTs) are presented. Multi-walled and double-walled carbon nanotubes were used. Penetration enhancement into the skin following passive diffusion and iontophoresis were determined. Possible solubility enhancement effects and drug adsorption properties of CNTs were investigated. CNTs were found to be a useful drug carrier system and they can provide a high loading and enhanced transdermal penetration for especially hydrophobic drugs. The electroconductive nature of CNTs allows easy application of iontophoresis and the additional advantage of CNTs appears to be using them as electrodes.


The rise of breastfeeding in the United States.
What factors influenced the resurgence of breastfeeding in the last decades of the twentieth century? This article has considered several explanations. Demographic trends, particularly the increased birth rate among black and Hispanic women, coupled with the resurgence of breastfeeding in these groups, may have contributed to the increase in the breastfeeding rate during the 1990s but likely played a minimal role in the earlier, more dramatic increase. The decrease in breastfeeding in the earlier part of the twentieth century may be partly attributable to increased maternal employment, but the resurgence of breastfeeding occurred during the late twentieth century--a period of unprecedented influx of new mothers into the workforce. There is no evidence that health care practitioners are providing more support for breastfeeding, and most international and US policies postdated the resurgence of breastfeeding, although they may have influenced the increase in the 1990s. A more plausible explanation of the resurgence of breastfeeding in all major segments of society is the pervasive influence of the natural-childbirth movement of the 1960s and 1970s, with its effects on the standard management of childbirth. Also, the increase in breastfeeding among low-income women may be attributable partly to programmatic changes in the provision of supplemental food through the WIC program and the targeting of breastfeeding-promotion efforts to the specific concerns of these women. Although breastfeeding increased at the end of the twentieth century relative to earlier decades, the disparity between the recommended rates and those achieved by US women is great. Thus, efforts to increase breastfeeding initiation and duration should continue, particularly for the groups that are at greatest risk for illness, such as minority and low-income infants. This article suggests that the strategies likely to have a lasting effect on future breastfeeding rates will be social pressures that affect existing barriers to breastfeeding. Such pressures may come from health maintenance organizations, insurance companies, and the US government, which are likely to increasingly recognize the costs of not breastfeeding to their institutions. The provision of flexible work hours and paid maternity leave, either by the US government or family-friendly workplaces, could increase the ability of employed women to optimally feed their infants. As Retsinas noted in an article on the cultural context of breastfeeding, "While it is 'known' that breastfeeding is better, our society is not structured to facilitate that choice." Efforts to improve breastfeeding rates need to make visible the wider cultural context in which infant-feeding choices are made and alter components that make it difficult for US women to feed their infants optimally.


Fully Integrated Oncology and Palliative Care Services at a Local Hospital in Mid-Norway: Development and Operation of an Innovative Care Delivery Model.
Early access to cancer palliative care is recommended. Descriptions of structures and processes of outpatient palliative care clinics operated within smaller hospitals are scarce. This paper presents the development and operation of a fully integrated cancer and palliative care outpatient clinic at a local hospital in a rural region of Mid-Norway offering palliative care concurrent with cancer treatment. A standardized care pathway was applied. Palliative care is in Norway part of the public healthcare system. Official recommendations recent years point out action points to improve delivery of palliative care. An integrated cancer and palliative care outpatient clinic at a local hospital and an innovative care delivery model was developed and operated in this setting. Patients were recruited for a descriptive study of the patient population. Clinical data were collected by clinical staff and 13 symptom intensities were reported by the patients. Cancer and palliative care were provided by one team of healthcare professionals trained in both fields. There was a close collaboration with the other departments at the hospital, with its affiliated tertiary hospital, and with community health and care services to provide timely referral, enhanced continuity, and improved coordination of care. Eighty-eight patients were included. Mean age was 65.6 years, the most common cancer diagnoses were digestive organs (22.7%), male genital organs (20.5%) or breast (25.0%), 75.0% had metastatic or locally advanced cancer, 59.1% were treated with non-curative intention and 93.1% had Karnofsky Performance Status ≥ 80%. Median scores of individual symptoms ranged from 0 to 3 (numerical rating scale, 0-10) and 61.0% reported at least one clinically significant symptom rating (≥ 4). This delivery model of integrated outpatient cancer and palliative care is particularly relevant in rural regions allowing cancer patients access to palliative care earlier in the disease trajectory and closer to home.


Massive subretinal and vitreous haemorrhages at presentation in idiopathic thrombocytopenic purpura: report of a case and review of literature.
A 21-year-old woman presented with acute decreased vision due to massive subretinal haemorrhage with macular exudation in the right eye and vitreous haemorrhage in the left eye. Haematological work-up revealed thrombocytopenia (platelets 14,000/μl) with anaemia (haemoglobin 6.3 gm/100 ml). A diagnosis of idiopathic thrombocytopenic purpura (ITP) was confirmed by a haematologist. Systemic therapy resulted in dramatic resolution of the fundus features with near complete restoration of visual acuity over three months. ITP can present with intraocular haemorrhages, especially if accompanied by anaemia. A high index of suspicion and appropriate systemic treatment can lead to a successful outcome.


Abbott monoclonal enzyme immunoassay measurement of estrogen receptors in human breast cancer: a European multicenter study.
A new enzyme immunoassay (Abbott ER-EIA Monoclonal) for the determination of estrogen receptor in cytosols from breast tumor specimens has been developed by Abbott Laboratories. To establish the correlation of the results from this new technique with currently existing steroid binding methods, a multicenter study was conducted in eight European laboratories. All participants followed the same protocol consisting of a familiarization phase, a proficiency evaluation, and a comparison of existing steroid binding methods with the new immunoassay using panel samples and clinical specimens. ER-EIA was compared with the multipoint dextran coated charcoal assay in six laboratories, four of which followed the EORTC protocol; of the remaining two laboratories, one used a single saturating dose assay, the other an isoelectric focusing assay. The results show no significant difference between reducing agents when used in the ER-EIA. Reproducibility for the immunoassay (interassay coefficient of variation, 6%, interlaboratory coefficient of variation, 11-19%) was somewhat better than that for the steroid binding methods (interlaboratory coefficient of variation, 12-32%). The correlation between the methods was dependent on the origin of the lyophilized specimens. In breast tumor samples, an excellent correlation, (not statistically different from 1) was found between the ER-EIA and the steroid binding method in six laboratories. One laboratory showed a slope of 1.1 for the correlation line; the laboratory using isoelectric focusing showed a slope of 1.9. The mean value determined by the enzyme immunoassay in premenopausal women was 74 fmol/mg cytosol protein, and in postmenopausal women it was 187 fmol/mg cytosol protein with no significant difference in the slope of the correlation line. Results suggest the usefulness of the new standardized enzyme immunoassay for routine use in the clinical laboratory.


Acetylglycerylether phosphorylcholine-(AGEPC) and leukotriene B4-stimulated cyclic AMP levels in human polymorphonuclear leukocytes.
It seems paradoxical that AGEPC induces a transient rise in cyclic AMP, yet the preincubation of neutrophils with agents that elevate cyclic AMP actually inhibits AGEPC-induced aggregation. However, similar transient elevations in cyclic AMP are observed using other stimulators of PMN function such as fmet-leu-phe, C5a (22), immune complexes (24), and phagocytosable particles (11). Elevations in cyclic AMP by PGE1, PGI2, dibutyryl cyclic AMP, and phosphodiesterase inhibitors, before the addition of an agonist, also blocked subsequent neutrophil activation in the above studies. Thus, these observations are not unique to AGEPC. However, the finding that the cyclooxygenase inhibitor indomethacin enhanced AGEPC-stimulated cyclic AMP accumulation is a novel observation and suggests that some oxygenated derivative of the 5-lipoxygenase pathway is responsible for the increase in cyclic AMP. The evidence for the association of the spike in cyclic AMP and the 5-lipoxygenase is strengthened by the observation that the 5-lipoxygenase inhibitor U-60257 attenuates the AGEPC-induced spike in cyclic AMP. It should be noted that U-60257 does not antagonize LTB4-stimulated cyclic AMP accumulation and has no direct influence on the neutrophil adenylate cyclase. The final correlation of the spike in cyclic AMP and the 5-lipoxygenase is made by the fact that LTB4 itself stimulates cyclic AMP levels in intact neutrophils as well as the adenylate cyclase in cell homogenates. As is the case with AGEPC, the transient spike in cyclic AMP induced by LTB4 is coincident with the onset of neutrophil aggregation. However, it is clear that the spike in neutrophil cyclic AMP induced by AGEPC can be dissociated from neutrophil aggregation.(ABSTRACT TRUNCATED AT 250 WORDS)


The use of Eugenol and electro-narcosis as anaesthetics: transcriptional impacts on the European eel (Anguilla anguilla L.).
Ecotoxicological studies aim to assess the potential environmental risks of various products. This implies the use of various biological models and tests on live animals. In case of handling fish and mammals, ethical rules have to be respected. The use of anaesthesia is considered to be the best way to ensure animal welfare. Eugenol and electro-narcosis are among the most popular chemical and physical anaesthetics used in fisheries and by field biologists. In this study, the genetic and endocrine impacts of these anaesthetics were assessed in order to establish whether the use of such methods could skew the results of ecotoxicological studies. Twenty yellow European eels (Anguilla anguilla) were submitted to Eugenol (50mg/L) and electro-narcosis until they reached a level of deep anaesthesia, while 20 other eels were kept aware. Five anaesthetized and five unanaesthetized eels were sacrificed and analysed directly after treatment and after 1, 7 and 21 days of recovery. At the brain level, Eugenol triggered an increase in the transcription level of genes encoding proteins involved in oxidative stress responses (catalase expression 2.5-fold, mitochondrial superoxide dismutase expression 3-fold), probably due to a hypoxic event during anaesthesia. Later impacts were detected in muscles 21 days after anaesthesia (ATP synthase subunit 6 3-fold, NADH deshydrogenase subunit 5 4-fold and mitochondrial superoxide dismutase 3-fold increased) revealing oxidative stress from an accrued mitochondrial respiratory metabolism. Hormone dosages showed that the use of Eugenol reduced the release of plasma cortisol during anaesthesia. However, this impact seemed to be reversible within one day. In case of electro-narcosis, no significant variation in transcriptional levels could be detected between anaesthetized and unanaesthetized eels. Our results suggest that the use of Eugenol as an aesthetic in ecotoxicological studies measuring gene expression or plasma cortisol concentration is not appropriate, while electro-narcosis does not seem to have any impact, at least on the parameters taken into consideration in this study.


Evaluation of the protected brush catheter and bronchoalveolar lavage in the diagnosis of nosocomial pneumonia.
We assess the sensitivity and specificity of the protected brush catheter (PBC) and bronchoalveolar lavage (BAL) in diagnosing nosocomial pneumonia in nonimmunocompromised critically ill patients. Computerized bibliographic literature searches of MEDLINE were performed, and the reference list of each article selected was reviewed. Of 496 citations, there were 19 articles (describing 18 studies) that proved relevant. Study quality was assessed, and descriptive information concerning study populations, interventions, and clinically relevant outcome measurements was extracted. The sensitivity and specificity of PBC were high (pooled estimates, 89.9 and 94.5%, respectively). Criteria for a positive BAL have varied between studies, and sensitivity ranged from 53.3 to 100%, whereas specificity was 98.6%. Most studies did not report whether antibiotics were withheld on the basis of negative test results. In those that did, the incidence of adverse outcomes consequent on withholding antibiotics was low. BAL and PBC, combined with the use of quantitative cultures, appear to increase accuracy in diagnosing pneumonia. The strength of inference is hampered, however, by the absence of a "gold standard" for the diagnosis of pneumonia. Moreover, the generalizability of these findings is limited by the fact that there are so few methodologically sound studies from so few centers. A randomized trial of PBC is needed.


Genetic aspects of fibrodysplasia ossificans progressiva.
Complete ascertainment of fibrodysplasia ossificans progressiva in the United Kingdom was attempted and 44 patients were identified. This indicates a point prevalence of 0.61 x 10(-6). The disease is determined as an autosomal dominant trait which has complete penetrance but variable expressivity. No evidence for genetic heterogeneity was found in this series. All patients represented fresh gene mutations and their biological fitness was zero. Geographical clustering of these new mutations was evident but conformed to the general population distribution. The direct estimate of the mutation rate was 1.8 (SE +/- 1.04) x 10(-6) mutations per gene per generation. A significant paternal age effect was evident for these new mutations in the United Kingdom.


Guidelines for future research in constraint-induced movement therapy for children with unilateral cerebral palsy: an expert consensus.
The aim of this study was to provide an overview of what is known about constraintinduced movement therapy (CIMT) in children with unilateral cerebral palsy (CP), to identify current knowledge gaps, and to provide suggestions for future research. Nine experts participated in a consensus meeting. A comprehensive literature search was conducted and data were summarized before the meeting. The core model produced by the European network for Health Technology Assessment was used as a framework for discussion and to identify critical issues for future research. All models of CIMT have demonstrated improvements in the upper limb abilities of children with unilateral CP. A consensus was reached on 11 important questions to be further explored in future studies. The areas of highest priority included the effect of dosage, the effect of repeated CIMT, and the impact of predictive factors, such as age, on the response to CIMT. Consensus suggestions for future study designs and the use of validated outcome measures were also provided. The CIMT construct is complex, and much remains unknown. It is unclear whether a specific model of CIMT demonstrates superiority over others and whether dosage of training matters. Future research should build upon existing knowledge and aim to provide information that will help implement CIMT in various countries with different healthcare resources and organizational structures.


Decreased expression of long noncoding RNA GAS5 indicates a poor prognosis and promotes cell proliferation in gastric cancer.
Gastric cancer is the second leading cause of cancer death and remains a major clinical challenge due to poor prognosis and limited treatment options. Long noncoding RNAs (lncRNAs) have emerged recently as major players in tumor biology and may be used for cancer diagnosis, prognosis, and potential therapeutic targets. Although downregulation of lncRNA GAS5 (Growth Arrest-Specific Transcript) in several cancers has been studied, its role in gastric cancer remains unknown. Our studies were designed to investigate the expression, biological role and clinical significance of GAS5 in gastric cancer. Expression of GAS5 was analyzed in 89 gastric cancer tissues and five gastric cancer cell lines by quantitative reverse-transcription polymerase chain reaction (qRT-PCR). Over-expression and RNA interference (RNAi) approaches were used to investigate the biological functions of GAS5. The effect of GAS5 on proliferation was evaluated by MTT and colony formation assays, and cell apoptosis was evaluated by hochest stainning. Gastric cancer cells transfected with pCDNA3.1 -GAS5 were injected into nude mice to study the effect of GAS5 on tumorigenesis in vivo. Protein levels of GAS5 targets were determined by western blot analysis. Differences between groups were tested for significance using Student's t-test (two-tailed). We found that GAS5 expression was markedly downregulated in gastric cancer tissues, and associated with larger tumor size and advanced pathologic stage. Patients with low GAS5 expression level had poorer disease-free survival (DFS; P = 0.001) and overall survival (OS; P < 0.001) than those with high GAS5 expression. Further multivariable Cox regression analysis suggested that decreased GAS5 was an independent prognostic indicator for this disease (P = 0.006, HR = 0.412; 95%CI = 2.218-0.766). Moreover, ectopic expression of GAS5 was demonstrated to decrease gastric cancer cell proliferation and induce apoptosis in vitro and in vivo, while downregulation of endogenous GAS5 could promote cell proliferation. Finally, we found that GAS5 could influence gastric cancer cells proliferation, partly via regulating E2F1 and P21 expression. Our study presents that GAS5 is significantly downregulated in gastric cancer tissues and may represent a new marker of poor prognosis and a potential therapeutic target for gastric cancer intervention.


The Predictive Validity of Four Intelligence Tests for School Grades: A Small Sample Longitudinal Study.
Intelligence is considered the strongest single predictor of scholastic achievement. However, little is known regarding the predictive validity of well-established intelligence tests for school grades. We analyzed the predictive validity of four widely used intelligence tests in German-speaking countries: The Intelligence and Development Scales (IDS), the Reynolds Intellectual Assessment Scales (RIAS), the Snijders-Oomen Nonverbal Intelligence Test (SON-R 6-40), and the Wechsler Intelligence Scale for Children (WISC-IV), which were individually administered to 103 children (Mage = 9.17 years) enrolled in regular school. School grades were collected longitudinally after 3 years (averaged school grades, mathematics, and language) and were available for 54 children (Mage = 11.77 years). All four tests significantly predicted averaged school grades. Furthermore, the IDS and the RIAS predicted both mathematics and language, while the SON-R 6-40 predicted mathematics. The WISC-IV showed no significant association with longitudinal scholastic achievement when mathematics and language were analyzed separately. The results revealed the predictive validity of currently used intelligence tests for longitudinal scholastic achievement in German-speaking countries and support their use in psychological practice, in particular for predicting averaged school grades. However, this conclusion has to be considered as preliminary due to the small sample of children observed.