The Effect of Thymoquinone on Oxidative Stress Parameters and Apolipoprotein E in Alzheimer Model in Rats.
In this study, it was aimed to investigate the effect of thymoquinone (TQ) on oxidative stress and apolipoprotein E (ApoE) in an experimental Alzheimer's model created with AlCl3 and D-galactose in rats. Thirty-six Wistar Albino male rats saline group (Group 1), aluminum chloride (AlCl3) + D-galactose (D-Gal) group (Group 2), AlCl3 + D-Gal + TQ group (Group 3) were divided into 3 groups. The study was completed with 33 rats. Group 1 was given saline intraperitoneally (i.p) for 28 days. 2nd group; D-Gal at a dose of 60 mg/kg/day and AlCl3 at a dose of 40 mg/kg/day were given i.p. daily for 28 days. 3rd group; D-Gal at a dose of 60 mg/kg/day and AlCl3 at a dose of 40 mg/kg/day were given i.p. daily for 28 days. Group 3 rats were given 20 mg/kg/day TQ in corn oil by gavage for 14 days. Malonyl dialdehyde (MDA), superoxide dismutase (SOD), total antioxidant capacity (TAS), total oxidant capacity (TOS), glutathione peroxidase (GsH-Px), and ApoE levels were determined in the blood and brain tissues of rats in all three groups. One-way ANOVA test was used in the statistical analysis of the data. Means of TAS, TOS, GSH-Px, SOD, MDA, and ApoE in blood and brain tissue of all three groups (excluding ApoE in brain tissue) were different from each other and this difference was statistically significant (p < 0.05). In this study, TQ, it was determined that all oxidative stress parameters examined had positively affected and decreased blood tissue ApoE levels. TQ can be used as an antioxidant and curative in Alzheimer's disease.


H60: A Unique Murine Hematopoietic Cell-Restricted Minor Histocompatibility Antigen for Graft-versus-Leukemia Effect.
Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is an important treatment for many types of hematological malignancies. Matching of donor and recipient for the major histocompatibility complex (MHC) improves the HSCT reconstitution, but donor-derived T cells reactive to non-MHC encoded minor histocompatibility antigens (MiHAs) can induce graft-versus-host disease (GVHD) while also being needed for graft-versus-leukemia (GVL) effects. MiHAs are allelically variant self-peptides presented conventionally on MHC molecules, but are alloantigenic in transplantation settings. Immunodominant MiHAs are most strongly associated with GVHD and GVL. There is need for mouse paradigms to understand these contradictory effects. H60 is a highly immunodominant mouse MiHA with hematopoietic cell-restricted expression. Immunodominance of H60 is tightly associated with its allelic nature (presence vs. absence of the transcripts), and the qualitative (TCR diversity) and quantitative (frequency) traits of the reactive T cells. The identity as a hematopoietic cell-restricted antigen (HRA) of H60 assists the appearance of the immunodominace in allo-HSCT circumstances, and generation of GVL effects without induction of serious GVHD after adoptive T cell transfer. Also it allows the low avidity T cells to escape thymic negative selection and exert GVL effect in the periphery, which is a previously unevaluated finding related to HRAs. In this review, we describe the molecular features and immunobiology in detail through which H60 selectively exerts its potent GVL effect. We further describe how lessons learned can be extrapolated to human allo-HCST.


QM/MM studies of the electronic structure of the compound I intermediate in cytochrome c peroxidase and ascorbate peroxidase.
Cytochrome c peroxidase (CcP) and ascorbate peroxidase (APX) both involve reactive haem oxoferryl intermediates known as 'compound I' species. These two enzymes also have a very similar structure, especially in the vicinity of the haem group. Despite this similarity, the electronic structure of compound I in the two enzymes is known to be very different. Compound I intermediates have three unpaired electrons, two of which are always situated on the Fe-O core, whilst the third is located in a porphyrin orbital in APX and many other compound I species. In CcP, however, this third unpaired electron is positioned on a tryptophan residue lying close to the haem ring. The same residue is present in the same position in APX, yet it is not oxidized in that case. We report QM/MM calculations, using accurate B3LYP density functional theory for the QM region, on the active intermediate for both enzymes. We reproduce the observed difference in electronic structure, and show that it arises as a result of subtle electrostatic effects which affect the ionization potential of both the tryptophan and porphyrin groups. The computed structures of both enzymes do not involve deprotonation of the tryptophan group, or protonation of the oxoferryl oxygen.


[Methods for in vitro antifungal susceptibility testing].
During the last years, a large amount of work has been completed to improve the methods used for in vitro antifungal susceptibility testing. Reference techniques are currently available both for yeasts and filamentous fungi, but in some instances, technical improvement are needed. Etest is another well standardized method that can be used as an alternative on a routine basis in the clinical microbiology laboratory. Studies of in vitro-in vivo correlations have led to the definition of susceptibility breakpoints for yeasts for fluconazole, itraconazole, and flucytosine.


Event-recorder monitoring in the diagnosis of atrial fibrillation in symptomatic patients: subanalysis of the SOPAT trial.
In the SOPAT (suppression of paroxysmal atrial tachyarrhythmias) trial, a high number of asymptomatic atrial fibrillation (AF) episodes were registered in patients with symptomatic AF. This subanalysis was performed to answer three questions: (1) Does antiarrhythmic therapy influence the ratio of symptomatic to asymptomatic AF? (2) Are symptoms during AF dependent on the heart rate (HR)? (3) Do symptoms correlate with an episode of AF? Within 60 months 1,033 patients with symptomatic AF were randomized to either quinidine plus verapamil (Q+V) 480/240 mg/day, Q+V 320/160 mg/day, sotalol 320 mg/day, or placebo, and followed up by daily and symptom-triggered ECG event recording. Over 188,634 ECGs were analyzed (87% SR, 7% AF). Symptoms were reported in only 46% of AF-ECGs. Quinidine plus verapamil reduced the ratio of symptomatic to asymptomatic AF, whereas sotalol had no effect (median [%] (25/75% quartile): Q+V 480/240: 33 (0/79), Q+V 320/160: 45 (1/82), sotalol: 56 (7/93), placebo: 63 (8/92)). The HR during AF directly correlates with the occurrence of symptoms (P < 0.001) and was significantly lower during asymptomatic AF in all groups (mean +/- SD: sympt.: 113 +/- 27/minute, asympt.: 103 +/- 27/minute, P < 0.001). Both antiarrhythmics reduced the mean HR compared to placebo (P < 0.001). Analyzing all symptom-triggered ECGs, AF was diagnosed in only 37%, whereas the majority were SR. Taken together, in contrast to sotalol, Q+V reduces the ratio of symptomatic to asymptomatic AF compared with placebo, at least in part by decreasing the heart rate. Furthermore, patients' symptoms are not a reliable surrogate parameter for the prevalence of AF.


Evaluation of Intracameral Pentablock Copolymer Thermosensitive Gel for Sustained Drug Delivery to the Anterior Chamber of the Eye.
To investigate PTSgels (Pentablock copolymers) as an injectable formulation technology for sustained ocular drug delivery. Drug release profile, tolerability, and polymer degradation for one of the thermosensitive, biodegradable, and biocompatible compositions were investigated through intracameral (IC) injection in rabbits. New Zealand White rabbit eyes were injected IC (50 μL) with 100 μg near-infrared-immunoglobulin G (NIR-IgG) in balanced salt solution (BSS) or 20% PTSgel; or with PTSgel or BSS alone. Ocular irritation scoring, intraocular pressure (IOP), and corneal thickness (CT) measurement, as well as color and infrared photography, were performed for up to 28 days postinjection. Upon euthanasia at 7, 14, or 28 days, eyes underwent ex vivo imaging (Xenogen IVIS) followed by tissue fixation and histopathology. IC injection of PTSgel (liquid at room temperature) was performed without difficulty using a 31G needle. The polymer quickly gelled in the IC space resulting in an inferior anterior chamber deposit. The tested PTSgel was well tolerated, with no significant changes in IOP or CT. Eyes injected with NIR-IgG in PTSgel had visible NIR-IgG through 9 days postinjection, and ex vivo imaging detected a strong NIR-IgG signal in the anterior chamber through day 28. The gel deposit steadily decreased in size over time and was nearly eliminated by 28 days. The PTSgel released IgG for 28 days and was well tolerated. The polymer degraded in parallel with drug release. These results demonstrate the potential of intracameral PTSgel formulations for sustained delivery of biologic therapies to the ocular anterior segment.


Conditioned inhibition in preweanling rats.
Inhibitory conditioning is a very well established phenomenon in associative learning that has been demonstrated in both humans and adult animals. But in spite of the fact that this topic has generated much empirical and theoretical work, there are no published studies assessing inhibitory learning during the early ontogeny of the rat. In this study we test the possibility of finding conditioned inhibition in infant rats (Day 10) using a conditioned taste aversion procedure. We tested whether the consumption of saccharin (A) was reduced when paired with a LiCl injection compared to the presentation of saccharin in compound with a lemon odor (AX) without any aversive consequence. After training, retardation, and summation tests were conducted in order to evaluate the inhibitory properties of the lemon odor (X). The results of this study showed that in male pups, after conditioned inhibition training, stimulus X passed both retardation and summation tests. These results indicate that conditioned inhibition can be established in the early development of the rat, suggesting that animals at this stage of ontogeny have the capacity to acquire and to express inhibitory conditioning, although this effect appears to be sex-dependent.


How large are nonadiabatic effects in atomic and diatomic systems?
With recent developments in simulating nonadiabatic systems to high accuracy, it has become possible to determine how much energy is attributed to nuclear quantum effects beyond zero-point energy. In this work, we calculate the non-relativistic ground-state energies of atomic and molecular systems without the Born-Oppenheimer approximation. For this purpose, we utilize the fixed-node diffusion Monte Carlo method, in which the nodes depend on both the electronic and ionic positions. We report ground-state energies for all systems studied, ionization energies for the first-row atoms and atomization energies for the first-row hydrides. We find the ionization energies of the atoms to be nearly independent of the Born-Oppenheimer approximation, within the accuracy of our results. The atomization energies of molecular systems, however, show small effects of the nonadiabatic coupling between electrons and nuclei.


Integrated Bioactive Scaffold with Polydeoxyribonucleotide and Stem-Cell-Derived Extracellular Vesicles for Kidney Regeneration.
Kidney tissue engineering and regeneration approaches offer great potential for chronic kidney disease treatment, but kidney tissue complexity imposes an additional challenge in applying regenerative medicine for renal tissue regeneration. In this study, a porous pneumatic microextrusion (PME) composite scaffold consisting of poly(lactic-co-glycolic acid) (PLGA, P), magnesium hydroxide (MH, M), and decellularized porcine kidney extracellular matrix (kECM, E) is functionalized with bioactive compounds, polydeoxyribonucleotide (PDRN), and tumour necrosis factor-α (TNF-α)/interferon-γ (IFN-γ)-primed mesenchymal stem-cell-derived extracellular vesicles (TI-EVs) to improve the regeneration and maintenance of a functional kidney tissue. The combination of PDRN and TI-EVs showed a significant synergistic effect in regenerative processes including cellular proliferation, angiogenesis, fibrosis, and inflammation. In addition, the PME/PDRN/TI-EV scaffold induced an effective glomerular regeneration and restoration of kidney function compared to the existing PME scaffold in a partial nephrectomy mouse model. Therefore, such an integrated bioactive scaffold that combines biochemical cues from PDRN and TI-EVs and biophysical cues from a porous PLGA scaffold containing MH and kECM can be used as an advanced tissue engineering platform for kidney tissue regeneration.


Transcatheter closure of complex iatrogenic ventricular septal defect: a case report.
Iatrogenic membranous ventricular septal defects (VSDs) are rare complications of cardiothoracic surgery, such as septal myectomy for hypertrophic obstructive cardiomyopathy (HOCM). Transcatheter closure is considered an appealing alternative to surgery, given the increased mortality associated with repeated surgical procedures, but reports are extremely limited. We herein report the case of a 63-year-old woman with HOCM who underwent successful percutaneous closure of an iatrogenic VSD after septal myectomy. Two percutaneous techniques are discussed, namely the 'muscular anchoring' and the 'buddy wire delivery', aimed at increasing support and providing stability to the system during percutaneous intervention. Transcatheter closure represents an attractive minimally invasive approach for the management of symptomatic iatrogenic VSDs. The new techniques described could help operators to cross tortuous and tunnelled defects and to deploy closure devices in case of complex VSD anatomy.


[Xanthan production by Xanthomonas campestris B-1459].
The production of xantano from Xanthomonas campestris B-1459 was analyzed. The experiments were performed in shaked flasks at 250 rpm and 2.5 cm eccentricity. Using a base modified medium it was possible to achieve xantano concentration of 35 g/l in 72 h of process. The modified medium contained glucose as carbon source, and yeast extract, meat peptone, malt extract and amaranth meal as growth factors and nitrogen sources, in a KH2PO4/K2HPO4 buffer.


Multifunctional Hydrogel with Good Structure Integrity, Self-Healing, and Tissue-Adhesive Property Formed by Combining Diels-Alder Click Reaction and Acylhydrazone Bond.
Hydrogel, as a good cartilage tissue-engineered scaffold, not only has to possess robust mechanical property but also has to have an intrinsic self-healing property to integrate itself or the surrounding host cartilage. In this work a double cross-linked network (DN) was designed and prepared by combining Diels-Alder click reaction and acylhydrazone bond. The DA reaction maintained the hydrogel's structural integrity and mechanical strength in physiological environment, while the dynamic covalent acylhydrazone bond resulted in hydrogel's self-healing property and controlled the on-off switch of network cross-link density. At the same time, the aldehyde groups contained in hydrogel further promote good integration of the hydrogel to surrounding tissue based on aldehyde-amine Schiff-base reaction. This kind of hydrogel has good structural integrity, autonomous self-healing, and tissue-adhesive property and simultaneously will have a good application in tissue engineering and tissue repair field.


Autologous versus allogeneic unrelated donor transplantation for acute lymphoblastic leukemia: comparative toxicity and outcomes.
For patients with high-risk or relapsed acute lymphoblastic leukemia (ALL) lacking a related histocompatible donor, autologous (Auto) and unrelated donor (URD) transplantation are available options. We compared outcomes and toxicities in 712 patients with ALL (517 URD, 195 Auto) in first complete remission (CR1) or second complete remission (CR2) who underwent transplantation. All patients were <50 years old, although URD patientswere younger (median age, 14 versus 18 years, P < .002). The proportion of patients in CR1 versus CR2 was similar (36% versus 38%, P = .57), but more URD recipients than Auto recipients had high-risk karyotypes (25% versus 13%, P = .003) and white blood cell (WBC) counts > or =50 x 10(9)/L (33% versus 14%, P < .001). Engraftment was similar in URD and Auto recipients. Ex vivo purging delayed but did not prevent engraftment after Auto transplantation. Transplantation-related mortality was higher after URD transplantation (42%+/-8%) than after Auto transplantation (20%+/-12%) in CR1 (P = .004) and also in CR2. Conversely, relapse was more frequent after Auto transplantation in CR1 (Auto, 49%+/-12% versus URD, 14%+/-5%) and CR2 (64%+/-8% versus 25%+/-5%) (P < .0001). These findings showed net similar outcomes for these 2 transplantation choices. Transplantation in CR1 yielded similar 3-year survival rates for URD (51%+/-7%) and Auto (44%+/-12%), as did transplantation in CR2 (40%+/-6% versus 32%+/-9%, respectively). Multivariate regression analysis identified significantly better disease-free survival after the first 6 months in matched URD versus Auto in younger patients, in those in CR2 with CR1 >1year, WBC <50 x 10(9)/L, performance status > or =90%, and in those who have undergone transplantation since 1995. These comparative data suggest that both matched URD and Auto transplantation can yield extended survival. Although URD transplantation offers substantially better protection against leukemic relapse, improvements in allotransplantation safety and refinements in patient selection are required to better aid treatment decision making for the best overall survival.


Follow-up after radical surgery for colorectal cancer. Design of a randomized study.
The value of different follow-up examinations after radical surgery for colorectal cancer has not been proven. The risk of such programmes, including invasive examinations, may invalidate the possible benefit from early diagnosis of recurrent and metachronous cancer. The present trial is a randomized study, evaluating possible benefit from a very detailed programme compared to that of virtually no follow-up. The design is presented, but so far no more than 207 of the 600 patients wanted for trial, have been included. Differences in mortality rates, survival and morbidity will be evaluated and the influence of repeated polypectomy upon risk of metachronous colorectal cancer will also be estimated.


The effect of the Tylenol scare on parent's use of over-the-counter drugs.
We administered a questionnaire to determine changes in parental use and administration of over-the-counter (OTC) drugs after cyanide-laced Extra-Strength Tylenol capsules caused the deaths of at least seven people in Illinois in 1982. Three hundred area parents were studied and divided into three equally sized groups on the basis of economic, educational, and professional criteria. After the Tylenol murders, all groups became anxious about the safety of OTC drugs, but intergroup differences were highly significant (p = 0.001). Attitude changes, however, did not necessarily precipitate a change in behavior. Similar percentages of each of the groups who claimed to be adversely affected by the Tylenol scare gave OTC drugs as before.


Low-energy LED red light inhibits the NF-κB pathway and promotes hPDLSCs proliferation and osteogenesis in a TNF-α environment in vitro.
There are few studies on the effect of low-energy LED red light on periodontal tissue regeneration in an inflammatory environment. In this study, Cell Counting Kit-8 (CCK-8) assays were used to detect the effects of TNF-α at three different concentrations (0, 10 ng/ml, and 20 ng/ml) on the proliferation of human periodontal ligament stem cells (hPDLSCs), and 10 ng/ml was selected as the subsequent experimental stimulation concentration. CCK-8 assays were used to detect the effect of LED red light with energy density of 1 J/ cm2, 3 J/ cm2, and 5 J/cm2 on the proliferation of hPDLSCs. The promotion effect of energy density of 5 J/cm2 on the proliferation of hPDLSCS was the most obvious (p < 0.05). Set CON group, ODM group, ODM + 10 ng/ml TNF-α group, and ODM + 10 ng/ml TNF-α + 5 J/ cm2 LED red light group. Alkaline phosphatase staining and activity detection, alizarin red staining and calcium nodules quantitative detection of osteoblast differentiation products, real-time fluorescence quantitative PCR detection of osteoblast gene expression (Runx2, Col-I, OPN, OCN). The results showed that ODM showed the strongest osteoblast ability, followed by ODM + 10 ng/ml TNF-α + 5 J/ cm2 LED red light group. The osteoblast ability of ODM + 10 ng/ml TNF-α was decreased, but was not found in CON group. Western blot was used to detect the expression of NF-κB pathway protein and osteoblast-related proteins (Runx2, Col-I, OPN, OCN) after addition of PDTC inhibitor. The results showed that the expression of p-IκBα was increased and the expression of IκBα was decreased (p < 0.05). The expression of osteoblast protein increased after the addition of inhibitor (p < 0.05). Therefore, in an inflammatory environment constructed by 10 ng/ml TNF-α, 5 J/cm2 LED red light can upregulate the proliferation and osteogenesis of hPDLSCs by inhibiting NF-κB signaling pathway.


Y chromosome in Turner syndrome: detection of hidden mosaicism and the report of a rare X;Y translocation case.
Turner syndrome (TS) is a common genetic disorder in females associated with the absence of complete or parts of a second sex chromosome. In 5-12% of patients, mosaicism for a cell line with a normal or structurally abnormal Y chromosome is identified. The presence of Y-chromosome material is of medical importance because it results in an increased risk of developing gonadal tumours and virilisation. Molecular study and fluorescence in situ hybridisation approaches were used to study 74 Brazilian TS patients in order to determine the frequency of hidden Y-chromosome mosaicism, and to infer the potential risk of developing malignancies. Additionally, we describe one TS girl with a very uncommon karyotype 46,X,der(X)t(X;Y)(p22.3?2;q11.23) comprising a partial monosomy of Xp22.3?2 together with a partial monosomy of Yq11.23. The presence of cryptic Y-chromosome-specific sequences was detected in 2.7% of the cases. All patients with Y-chromosome-positive sequences showed normal female genitalia with no signs of virilisation. Indeed, the clinical data from Y-chromosome-positive patients was very similar to those with Y-negative results. Therefore, we recommend that the search for hidden Y-chromosome mosaicism should be carried out in all TS cases and not be limited to virilised patients or carriers of a specific karyotype.


Genome-Wide Identification and Functional Characterization of the Chloride Channel TaCLC Gene Family in Wheat (Triticum aestivum L.).
In plants, chloride channels (CLC) are involved in a series of specific functions, such as regulation of nutrient transport and stress tolerance. Members of the wheat Triticum aestivum L. CLC (TaCLC) gene family have been proposed to encode anion channels/transporters that may be related to nitrogen transportation. To better understand their roles, TaCLC family was screened and 23 TaCLC gene sequences were identified using a Hidden Markov Model in conjunction with wheat genome database. Gene structure, chromosome location, conserved motif, and expression pattern of the resulting family members were then analyzed. Phylogenetic analysis showed that the TaCLC family can be divided into two subclasses (I and II) and seven clusters (-a, -c1, -c2, -e, -f1, -f2, and -g2). Using a wheat RNA-seq database, the expression pattern of TaCLC family members was determined to be an inducible expression type. In addition, seven genes from seven different clusters were selected for quantitative real-time PCR (qRT-PCR) analysis under low nitrogen stress or salt stress conditions, respectively. The results indicated that the gene expression levels of this family were up-regulated under low nitrogen stress and salt stress, except the genes of TaCLC-c2 cluster which were from subfamily -c. The yeast complementary experiments illustrated that TaCLC-a-6AS-1, TaCLC-c1-3AS, and TaCLC-e-3AL all had anion transport functions for NO3 - or Cl-, and compensated the hypersensitivity of yeast GEF1 mutant strain YJR040w (Δgef1) in restoring anion-sensitive phenotype. This study establishes a theoretical foundation for further functional characterization of TaCLC genes and provides an initial reference for better understanding nitrate nitrogen transportation in wheat.


MRI of carotid atherosclerosis.
Although MRI is widely used to observe atherosclerosis impacts on the vessel lumen, MRI also depicts the size of the plaque itself, its composition, and plaque inflammation, providing information beyond simple stenosis. This article summarizes the state of evidence for a clinical role for MRI of carotid atherosclerosis. MRI of carotid atherosclerosis has a proven role in pharmaceutical trials and may improve patient management once large-scale clinical trials have been completed.


Embedding Cobalt Atom Clusters in CNT-Wired MoS2 Tube-in-Tube Nanostructures with Enhanced Sulfur Immobilization and Catalyzation for Li-S Batteries.
Lithium-sulfur batteries are one of the most promising next-generation energy storage systems. The efficient interconversion between sulfur/lithium polysulfides and lithium sulfide is a performance-determining factor for lithium-sulfur batteries. Herein, a novel strategy to synthesize a unique tube-in-tube CNT-wired sulfur-deficient MoS2 nanostructure embedding cobalt atom clusters as an efficient polysulfide regulator is successfully conducted in Li-S batteries. It is confirmed that encapsulating MWCNTs into hollow porous sulfur-deficient MoS2 nanotubes embedded with metal cobalt clusters not only can accelerate electron transport and confine the dissolution of lithium polysulfide by physical/chemical adsorption, but also can catalyze the kinetics of polysulfide redox reactions. Based on DFT calculations, in situ spectroscopic techniques, and various electrochemical studies, catalytic effects of CNT/MoS2 -Co nanocomposite in Li-S battery are deeply investigated for the first time. The CNT/MoS2 -Co composite cathode exhibits a very remarkable rate capability (641 mAh g-1 at 5.0 C) and excellent cycling stability (capacity decay rate of 0.050% per cycle at 5.0 C) even at high sulfur mass loading of 3.6 mg cm-2 . More crucially, CNT/MoS2 -Co tube-in-tube nanostructures present a superior specific capacity of 650 mAh g-1 in a Li-S pouch cell at 0.2 C (4.0 mg cm-2 ).


The effect of hair colour on anaesthetic requirements and recovery time after surgery.
Patients with red hair are much more likely to have a variant of the melanocortin-1 receptor gene and this may affect sensitivity to general anaesthetics and pain response. We did a prospective, matched cohort study of 468 healthy adult patients undergoing general anaesthesia for elective surgery. All patients received an inhalational general anaesthetic. Anaesthetic drugs and doses used, hypnotic depth, recovery times, pain scores and quality of recovery scores were recorded. More men than women had red hair, so we did subgroup and multivariable analyses to account for this imbalance. There was no significant difference in recovery times, pain scores or quality of recovery scores in those with red hair. After adjusting for age, sex, American Society of Anesthesiologists physical status and duration of surgery, the recovery ratio for time to eye-opening in redheads was comparable to those with black or brown hair, 0.82 (0.57-1.19), P=0.30. We found no evidence that patient hair colour affects anaesthetic requirements or recovery characteristics in a broad range of surgical procedures.


Cyclosporin associated headache.
Despite successful orthoptic liver transplantation some patients develop a recurrent headache that interferes with their quality of life. To estimate the frequency of this symptom 34 patients who had undergone orthoptic liver transplantation were questioned about the history and character of any headache. Six patients described a recurrent headache typical of migraine only since transplantation. In two patients the pain improved after reduction of cyclosporin dosage and thereby plasma cyclosporin concentration.


Defining the reference range for right ventricular systolic strain by echocardiography in healthy subjects: A meta-analysis.
Right ventricular (RV) systolic strain has recently demonstrated prognostic value in various cardiovascular diseases. Despite this, the reference range including the lower limit of normal (LLN) and factors associated with RV strain measurements are not well-established. This meta-analysis aimed to determine the mean and LLN of two- (2D) and three-dimensional (3D) right ventricular global (RVGLS), free wall (RVFWLS) and interventricular septal wall (IVSLS) longitudinal strains in healthy individuals and factors that affect strain measurements. In this meta-analysis, Pubmed, Embase and Cochrane databases were searched until 31 July 2020 for eligible studies reporting RVGLS, RVFWLS and/or IVSLS in at least 30 healthy subjects. We pooled the means and LLNs of RV strains by two- (2D) and three- (3D) dimensional echocardiography, and performed meta-regression analyses. From 788 articles screened, 45 eligible studies totaling 4439 healthy subjects were eligible for analysis. Pooled means and LLNs with 95% confidence intervals for 2D- RV strains were RVGLS -23.4% (-24.2%, -22.6%) and -16.4% (-17.3%, -15.5%) in 27 studies; RVFWLS -26.9% (-28.0%, -25.9%) and -18.0% (-19.2%, -16.9%) in 32 studies; and IVSLS -20.4% (-22.0%, -18.9%) and -11.5% (-13.6%, -9.6%) in 10 studies, and similar results for 3D- RV strains. Right ventricular fractional area change and vendor software were associated with 2D-RVGLS and RVFWLS means and LLNs. We reported the pooled means and LLNs of RV systolic strains in healthy subjects, to define thresholds for abnormal, borderline and normal strains. Important factors associated with RV systolic strains include right ventricular fractional area change and vendor software.


Heterogeneity in Lewis-X and sialyl-Lewis-X antigen expression on monocytes in whole blood: relation to stimulus-induced oxidative burst.
By using flow cytometric analysis of cells in whole blood expressing high levels of CD14, we found a subpopulation of monocytes (8% of total) with higher scatter parameters, high capacity to produce reactive oxygen species (ROS), stronger expression of Lewis-X (CD15), sialyl-Lewis-X, CD11b and CD18 antigens, as well as an increased polymerized actin content. The size of this subpopulation increased after stimulation with lipopolysaccharide at the expense of the remaining monocytes, suggesting that its features were inducible. The membrane increase in Lewis-X and sialyl-Lewis-X expression observed during this conversion was largely due to the translocation of these carbohydrate structures from intracellular pools. Moreover, this subpopulation behaved as a primed monocyte subpopulation producing large amounts of H2O2 in response to N-formyl-methionyl-leucyl-phenylalanine. Increased H2O2 production was inhibited not only by anti-CD14 but also by anti-CD15 and anti-sialyl-Lewis-X monoclonal antibodies when added before lipopolysaccharide. These results show that lipopolysaccharide priming is regulated, at least in part, by Lewis-X and sialyl-Lewis-X structures expressed on the monocyte membrane. All together, this highly reactive and inducible subpopulation of monocytes, which share phenotypic and functional characteristics with neutrophils, might play an important role in host defenses and inflammatory responses.


Role of Matrix Metalloproteinase-2 in the Development of Atherosclerosis among Patients with Coronary Artery Disease.
Coronary artery disease (CAD) is a caused by atherosclerotic plaque buildup in the coronary arteries that supply blood and oxygen to the heart. Matrix metalloproteinase (MMP) is a family of zinc-dependent endopeptidase that is involved in various stages of atherosclerosis as demonstrated in in vitro and in vivo studies. MMP-2 is associated with both stable and unstable atherosclerotic plaque formation. The current review aimed to identify the role of MMP-2 in atherosclerosis development among CAD patients. Literature search was conducted through four online databases and only studies that were published from 2018 until February 2023 were included. The risk of bias was assessed by using the Newcastle-Ottawa Scale. A total of 10,622 articles were initially identified, and only eight studies that fulfilled the selection criteria were included in this review. The results showed that MMP-2 levels and activity were higher in patients with unstable CAD than those with stable CAD and healthy subjects. There was a significant association between MMP-2 levels and cardiovascular disease with MMP-14 levels, which is a pro-MMP-2 activator. In addition, two single nucleotide polymorphisms of the MMP-2 gene (rs243865 and rs243866) were significantly associated with the development of atherosclerosis. In conclusion, MMP-2 plays a crucial role in the development of atherosclerosis among patients with CAD and could be a potential target for CAD therapy.


Artesunate inhibits RANKL-induced osteoclastogenesis and bone resorption in vitro and prevents LPS-induced bone loss in vivo.
Osteoclasts are multinuclear giant cells responsible for bone resorption in lytic bone diseases such as osteoporosis, arthritis, periodontitis, and bone tumors. Due to the severe side-effects caused by the currently available drugs, a continuous search for novel bone-protective therapies is essential. Artesunate (Art), the water-soluble derivative of artemisinin has been investigated owing to its anti-malarial properties. However, its effects in osteoclastogenesis have not yet been reported. In this study, Art was shown to inhibit the nuclear factor-κB ligand (RANKL)-induced osteoclastogenesis, the mRNA expression of osteoclastic-specific genes, and resorption pit formation in a dose-dependent manner in primary bone marrow-derived macrophages cells (BMMs). Furthermore, Art markedly blocked the RANKL-induced osteoclastogenesis by attenuating the degradation of IκB and phosphorylation of NF-κB p65. Consistent with the in vitro results, Art inhibited lipopolysaccharide (LPS)-induced bone resorption by suppressing the osteoclastogenesis. Together our data demonstrated that Art inhibits RANKL-induced osteoclastogenesis by suppressing the NF-κB signaling pathway and that it is a promising agent for the treatment of osteolytic diseases.


Chorea in systematic lupus erythematosus; a case report.
We report a 33-year-old female who developed a movement disorder during maintenance therapy for systemic lupus erythematosus (SLE). She was diagnosed with SLE at the age of 25, and experienced an episode of SLE-associated hemophagocytic syndrome at age 27, which was successfully treated with intensive immunosuppressive therapy. In November 2012, during maintenance therapy with prednisolone (PSL) 5 mg/day and tacrolimus 0.5 mg/day, she developed acute-onset involuntary movements that were classified as chorea in combination with athetosis in her right limbs and right homonymous hemianopia, which subsided after about 1 h. Her laboratory tests on admission showed an elevated serum anti-double- stranded DNA antibody, positive serum anti-cardiolipin IgG, and an elevated IgG index in cerebrospinal fluid. Magnetic resonance imaging (MRI) showed no significant abnormality on admission, but an ischemic change in her left pallidum appeared on day 7. She was treated with a combination of high-dose corticosteroid, immunosuppressive agents (rituximab, cyclophosphamide, mycofenolate mofetil), antithrombotic therapy (heparin, cilostazol), and dopamine antagonists. Her symptoms remitted partially. Chorea in SLE is recognized as an anti-phospholipid-antibody-associated disorder. In our case, both immunological and ischemic mechanisms were thought to be involved.


Improved anti-leukemia activities of adoptively transferred T cells expressing bispecific T-cell engager in mice.
Despite the impressive clinical efficacy of T cells engineered to express chimeric antigen receptors (CAR-Ts), the current applications of CAR-T cell therapy are limited by major treatment-related toxicity. Thus, safer yet effective alternative approaches must be developed. In this study, we compared CD19 bispecific T-cell engager (BiTE)-transferred T cells that had been transfected by RNA electroporation with CD19 CAR RNA-transferred T cells both in vitro and in an aggressive Nalm6 leukemia mouse model. BiTEs were secreted from the transferred T cells and enabled both the transferred and bystander T cells to specifically recognize CD19(+) cell lines, with increased tumor killing ability, prolonged functional persistence, increased cytokine production and potent proliferation compared with the CAR-T cells. More interestingly, in comparison with CD3/CD28 bead-stimulated T cells, T cells that were expanded by a rapid T-cell expansion protocol (REP) showed enhanced anti-tumor activities for both CAR and BiTE RNA-electroporated T cells both in vitro and in a Nalm6 mouse model (P<0.01). Furthermore, the REP T cells with BiTE RNAs showed greater efficacy in the Nalm6 leukemia model compared with REP T cells with CAR RNA (P<0.05) and resulted in complete leukemia remission.


Effect of adrenergic neuron inhibitors on the vascular response to sympathetic nerve stimulation.
In anesthetized cats, the lumbar sympathetic trunk was stimulated, and the responses of the resistance and capacitance vessels of the hindlimb were recorded (by resistography and plethysmography, respectively). The drugs used were reserpine, methyldopa and bretylium tosylate. It was found that reserpine and methyldopa inhibit the response of the resistance vessels to a greater extent than that of the capacitance vessels, while bretylium tosylate has a greater effect on the transmission of constrictor impulses to the capacitance vessels. Reserpine mainly suppresses responses to low-frequency stimulation and bretylium tosylate--those to high-frequency stimulation, while methyldopa affects them to the same degree at all frequencies.


[Relationship between myocardial remodeling and neurohumoral activation in patients with cardiac failure].
50 healthy subjects and 685 patients with heart failure (NYHA functional class I-IV) due to coronary heart disease (CHD) aged 34-74 years (mean age 48.1 +/- 8.22 years) were examined. Cardio-hemodynamics was assessed by M- and B-echocardiography from parasternal, subcostal and apical view on a short and long axis with transmitral Doppler. Plasma levels of epinephrine, norepinephrine, serotonin, aldosterone, STH, hydrocortisone were measured in the patients. Progression of cardiac failure is characterized by evolution of the left ventricular ellipse form into a ball shape primarily due to an increase in cross sectional cavity size. The changes of the left ventricular geometry in heart failure patients are combined with progressive reduction of the relative wall thickness index, intensification of the myocardial stress and impairment of diastolic function. The clearest intercoupling between myocardial remodeling and neurohumoral activation was registered in patients with asymptomatic heart failure. With growing severity of left ventricular dysfunction, plasm activity of serotonin, hydrocortisone and aldosterone increase more than levels of norepinephrine, epinephrine and STH.


Think before you flush! A sustainable aquatic eco-system's relation to human health.
What we do every day at work and in our home lives can make a difference in the quality of our environment. Consider, for example, the flushing of pharmaceuticals into the sewer system can lead to water pollution resulting in a threat to aquatic and human life. In contrast, keeping aquatic life healthy may contribute to human health. Some aquatic-based medications are currently on the market. Others are in various stages of development. In this article the authors argue that, for the benefit of both human and marine life, it is time to implement safer disposal methods for unwanted medications. The authors begin by sharing nursing's guiding principles for environmental health; after which they review research related to pharmaceutical pollution of water resources; describe health care treatments derived from marine life; and discuss suggestions for promoting aquatic health. They conclude that by taking care to preserve aquatic life, we contribute to the quality of our own human lives.


Checklist of the parasites of European eel Anguilla anguilla (Linnaeus, 1758) (Anguillidae) in Poland.
The present paper lists all parasite species of the European eel Anguilla anguilla (Linnaeus, 1758), recorded in Poland, in both its saltwater and freshwater habitats. The list has been drawn up, based on data acquired since 1844. The majority of included parasite species are presented with fish infection parameters together with data on their developmental stages and occupied microhabitats, localities and dates of collection of the eels themselves. The database includes 62 parasite taxa (including 50 species, nine identified to the genus level and three to higher taxa), representing at least 47 genera and 39 families. The most frequently-noted parasites of the European eel are the cestode Bothriocephalus claviceps, the nematodes Anguillicoloides crassus, Camallanus lacustris and Raphidascaris acus and the acanthocephalan Acanthocephalus lucii. Four alien species have been noted from this host: A. crassus, the monogeneans Pseudodactylogyrus anguillae and Pseudodactylogyrus bini and the acanthocephalan Paratenuisentis ambiguus. The present list includes both new host records and earlier records not included in previous lists of parasites of eels.


Engineering the flagellar type III secretion system: improving capacity for secretion of recombinant protein.
Many valuable biopharmaceutical and biotechnological proteins have been produced in Escherichia coli, however these proteins are almost exclusively localised in the cytoplasm or periplasm. This presents challenges for purification, i.e. the removal of contaminating cellular constituents. One solution is secretion directly into the surrounding media, which we achieved via the 'hijack' of the flagellar type III secretion system (FT3SS). Ordinarily flagellar subunits are exported through the centre of the growing flagellum, before assembly at the tip. However, we exploit the fact that in the absence of certain flagellar components (e.g. cap proteins), monomeric flagellar proteins are secreted into the supernatant. We report the creation and iterative improvement of an E. coli strain, by means of a modified FT3SS and a modular plasmid system, for secretion of exemplar proteins. We show that removal of the flagellin and HAP proteins (FliC and FlgKL) resulted in an optimal prototype. We next developed a high-throughput enzymatic secretion assay based on cutinase. This indicated that removal of the flagellar motor proteins, motAB (to reduce metabolic burden) and protein degradation machinery, clpX (to boost FT3SS levels intracellularly), result in high capacity secretion. We also show that a secretion construct comprising the 5'UTR and first 47 amino acidsof FliC from E. coli (but no 3'UTR) achieved the highest levels of secretion. Upon combination, we show a 24-fold improvement in secretion of a heterologous (cutinase) enzyme over the original strain. This improved strain could export a range of pharmaceutically relevant heterologous proteins [hGH, TrxA, ScFv (CH2)], achieving secreted yields of up to 0.29 mg L-1, in low cell density culture. We have engineered an E. coli which secretes a range of recombinant proteins, through the FT3SS, to the extracellular media. With further developments, including cell culture process strategies, we envision further improvement to the secreted titre of recombinant protein, with the potential application for protein production for biotechnological purposes.


Characterization of primary cutaneous CD8+/CD30+ lymphoproliferative disorders.
CD30 primary cutaneous lymphoproliferative diseases include both lymphomatoid papulosis (LyP) and primary cutaneous anaplastic large cell lymphoma (PCALCL). The neoplastic cell of most primary CD30 lymphoproliferative disorders is CD4 positive. The terminology LyP "type D" has been used to describe a growing number of cases of LyP with a predominantly CD8 infiltrate. PCALCL with a CD8 phenotype has also been described, which presents a particularly difficult diagnostic and management challenge, given the difficulty in distinguishing it histologically from other cytotoxic lymphomas such as primary cutaneous aggressive epidermotropic CD8 cytotoxic T-cell lymphoma and CD8 gamma/delta and natural killer/T-cell lymphoma. We report 7 additional cases of these rare cutaneous CD8/CD30 lymphoproliferative disorders. We also present a unique case of CD8/CD30 LyP with histologic similarities to LyP type B. In all 7 of our cases of CD8 LyP and CD8 anaplastic large cell lymphoma, we found focal to diffuse MUM-1 positivity. We propose that MUM-1 may represent an adjunctive marker for CD8 lymphoproliferative disease. Finally, we review the current literature on cases of CD8 LyP and PCALCL. For the 106 cases examined, we found similar clinical and histologic features to those reported for traditional CD4CD30 LyP and PCALCL.


[The intrafollicular pressure of tertiary follicles in relation to their diameter in the cow ovary].
In ovaries of 31 cows, which were in prooestrus, the diameters of all visible tertiary follicles and their intrafollicular pressure were measured. The pressure increased largely, if the diameter of the follicles was higher than 5 mm. This high intrafollicular pressure reached the filtering pressure of capillaries. So it is possible, that the peripheral portions of the theca interna from the follicles become ischaemic. The following metabolic acidosis activates the collagenolytic enzyme in the follicular wall. It is concluded that these enzymes could be active in the digestion of the follicle wall during ovulation. In that time the high intrafollicular pressure in spite of higher diameter (over 10 mm) slowly decreased.


Prediction of effective properties and sound absorption of random close packings of monodisperse spherical particles: Multiscale approach.
The transport and sound absorption properties of random close packings of monodisperse spherical particles are explored following a multiscale approach. First, the discrete element method is used to simulate the free fall of the monodisperse particles in a bounded domain to create virtual samples that are representative of real samples. Different particle diameters ranging from 1 to 16 mm are studied. From the virtual samples, representative volume elements (RVEs) are defined. Local partial differential equations governing the transport properties are numerically solved on the RVEs. From the discretized RVEs and the numerical solutions, eight transport properties (porosity, tortuosity, and viscous and thermal static tortuosities, permeabilities, and characteristic lengths) are derived. Micro-macro relationships between these properties and the particle diameter are developed. They are validated against experimental measurements of the open porosity and sound absorption coefficients. The relationships are used to analyze the salient sound absorption features of such media, notably the resonant sound absorption behavior. Expressions allowing identification of the optimal particle diameter for a given thickness, or conversely, the optimal thickness for a given particle diameter, for achieving 100% absorption at the first resonant absorption are derived.


Restoring Agency to the Human Actor.
A cursory read of the social psychological literature suggests that when people find themselves in strong situations, they fail to display agency. The early classic studies of conformity, obedience, and bystander intervention, for example, are renowned for showing that when challenged by strong situational pressures, participants acquiesced-even if it meant abandoning their moral principles or disregarding their own sensory data. Later studies of learned helplessness, ego depletion, and stereotype threat echoed this "power of the situation" theme, demonstrating that exposure to (or the expectation of) a frustrating or unpleasant experience suppressed subsequent efforts to actualize goals and abilities. Although this work has provided many valuable insights into the influence of situational pressures, it has been used to buttress an unbalanced and misleading portrait of human agency. This portrait fails to recognize that situations are not invariably enemies of agency. Instead, strong situational forces often allow for, and may even encourage, expressions of human agency. We examine the nature, causes, and consequences of this phenomenon. We endorse a broader approach that emphasizes how responding to situational pressure can coexist with agency. This new emphasis should create greater convergence between social psychological models and the experience of agency in everyday life.


Phosphoproteomic Analysis of Paper Mulberry Reveals Phosphorylation Functions in Chilling Tolerance.
Paper mulberry is a valuable woody species with a good chilling tolerance. In this study, phosphoproteomic analysis, physiological measurement, and mRNA quantification were employed to explore the molecular mechanism of chilling (4 °C) tolerance in paper mulberry. After chilling for 6 h, 427 significantly changed phosphoproteins were detected in paper mulberry seedlings without obvious physiological injury. When obvious physiological injury occurred after chilling for 48 h, a total of 611 phosphoproteins were found to be significantly changed at the phosphorylation level. Several protein kinases, especially CKII, were possibly responsible for these changes according to conserved sequence analysis. The results of Gene Ontology analysis showed that phosphoproteins were mainly responsible for signal transduction, protein modification, and translation during chilling. Additionally, transport and cellular component organization were enriched after chilling for 6 and 48 h, respectively. On the basis of the protein-protein interaction network analysis, a protein kinase and phosphatases hub protein (P1959) were found to be involved in cross-talk between Ca2+, BR, ABA, and ethylene-mediated signaling pathways. We also highlighted the phosphorylation of BpSIZ1 and BpICE1 possibly impacted on the CBF/DREB-responsive pathway. From these results, we developed a schematic for the chilling tolerance mechanism at phosphorylation level.


A widened substrate selectivity filter of eukaryotic formate-nitrite transporters enables high-level lactate conductance.
Bacterial formate-nitrite transporters (FNT) regulate the metabolic flow of small weak mono-acids derived from anaerobic mixed-acid fermentation, such as formate, and further transport nitrite and hydrosulfide. The eukaryotic Plasmodium falciparumFNT is vital for the malaria parasite by its ability to release the larger l-lactate substrate as the metabolic end product of anaerobic glycolysis in symport with protons preventing cytosolic acidification. However, the molecular basis for substrate discrimination by FNTs has remained unclear. Here, we identified a size-selective FNT substrate filter region around an invariant lysine at the bottom of the periplasmic/extracellular vestibule. The selectivity filter is reminiscent of the aromatic/arginine constriction of aquaporin water and solute channels regarding composition, location in the protein, and the size-selection principle. Bioinformatics support an adaptation of the eukaryotic FNT selectivity filter to accommodate larger physiologically relevant substrates. Mutations that affect the diameter at the filter site predictably modulated substrate selectivity. The shape of the vestibule immediately above the filter region further affects selectivity. This study indicates that eukaryotic FNTs evolved to transport larger mono-acid substrates, especially l-lactic acid as a product of energy metabolism.


Lesbians' attitudes and practices of cervical cancer screening: a qualitative study.
Cervical cancer is the third most prevalent cancer in women, and since the introduction of the Papanicolaou test (Pap test or Pap smear), the incidence of cervical cancer and mortality rates worldwide have declined substantially. However significant disparities have been identified between the cervical screening rates of heterosexual and lesbian women. This study explores the attitudes and practices that lesbians have towards cervical cancer screening and aims to identify why such disparities occur. A qualitative methodology based on feminist perspectives was used to collect narrative data from lesbians about their attitudes and practices of cervical screening through the use of semi structured interviews. Nine women who self-identified as lesbian that were living in New South Wales were recruited for the study. Interviews were digitally recorded and transcribed verbatim. Data were analysed using a thematic analysis approach. Four main themes emerged from the data namely: Encountering cervical cancer: "my friends had some early cancer cells detected", Misconceptions related to risk: "I am a lesbian I don't need one", Imposed screening: "It's a requirement of IVF treatment" and, Promoting cervical screening: "I think it should be spoken about in schools". Consistent with the literature, the findings show that the majority of these women do not undertake cervical screening at the recommended rate. This study highlights the multiple and complex issues related to cervical cancer screening for lesbians, mainly through misconceptions and underestimation of risk. Specific and targeted educational and promotional strategies are required for both lesbians and health professionals to enhance cervical cancer screening rates for lesbians in Australia.


Mitochondrial sequence variation in the Guahibo Amerindian population from Venezuela.
New data were obtained on mitochondrial DNA (mtDNA) from Guahibo from Venezuela, a group so far not studied using molecular data. A population sample (n = 59) was analyzed for mtDNA variation in two control-region hypervariable segments (HV1 and HV2) by sequencing. The presence or absence of a 9-bp polymorphism in the COII/tRNA(Lys) region was studied by direct amplification and electrophoretic identification. Thirty-eight variable sites were detected in regions HV1 and HV2, defining 26 mtDNA lineages; 23.7% of these were present in a single individual. The 9-bp deletion was found in 3.39% of individuals. Nucleotide and haplotype diversities were relatively high compared with other New World populations. The identified sequence haplotypes were classified into four major haplogroups (A-D) according to previous studies, with high frequencies for A (47.46%) and C (49.15%), low frequency for B (3.39%), and an absence of D.


Advanced oxidation protein products change biological behaviors of rat endometrial epithelial cells by activating ERK/P38 signaling pathways.
Advanced oxidation protein products (AOPPs) are a family of oxidized protein compounds and could induce oxidative stress and inflammatory lesion in various cells. The accumulation of AOPPs was associated with female reproductive diseases such as polycystic ovary syndrome (PCOS), leiomyoma and endometriosis. However, the relationship between AOPPs and endometrial cells is unclear. To explore the effects of accumulated AOPPs on endometrial cells, we treated normal rat endometrial epithelial cells (rEECs) and endometriosis model rats with AOPPs. Primary rEECs were collected from 8-week-old female Wistar rats. Increasing the amount of AOPPs in the media of rEECs enhanced rEEC proliferation and migration, and inhibited apoptosis. Moreover, AOPPs triggered the production of reactive oxygen species and nitrite along with activated ERK and P38 signal and this, in turn, led to an upregulation of proliferation and migration. With the treatment of antioxidants or the inhibitors of ERK and P38, the above effects of AOPPs on rEECs were attenuated. Additionally, in an endometriosis rat model, a similar phenomenon was observed in that the growth of endometriotic implants were promoted by AOPPs and EECs were significantly increased. This study indicated that the accumulation of AOPPs could promote rEEC proliferation and migration through ERK and P38 signal both in vivo and in vitroThis article has an associated First Person interview with the first author of the paper.


Restriction enzyme and Southern hybridization analyses of Pseudomonas aeruginosa strains from patients with cystic fibrosis.
Conventional typing schemes for Pseudomonas aeruginosa may not discriminate strains. P. aeruginosa isolates from patients with cystic fibrosis were examined by restriction enzyme analysis and Southern hybridization. There was marked diversity of restriction enzyme analysis patterns among P. aeruginosa DNAs from cystic fibrosis isolates; however, sequential isolates obtained from individual patients showed very little variation over an 8-year period. DNA fragments from the alginate biosynthesis gene complex, the exotoxin A gene, and the elastase gene of P. aeruginosa were used in Southern hybridization analysis. The patterns of hybridization to the elastase and algD gene probes were highly conserved in all isolates, therefore, these DNA fragments are not useful in discriminating strains, in contrast to the exotoxin A gene probe.


Imaging of cartilage and bone: promises and pitfalls in clinical trials of osteoarthritis.
Imaging in clinical trials is used to evaluate subject eligibility, and/or efficacy of intervention, supporting decision making in drug development by ascertaining treatment effects on joint structure. This review focusses on imaging of bone and cartilage in clinical trials of (knee) osteoarthritis. We narratively review the full-text literature on imaging of bone and cartilage, adding primary experience in the implementation of imaging methods in clinical trials. Aims and constraints of applying imaging in clinical trials are outlined. The specific uses of semi-quantitative and quantitative imaging biomarkers of bone and cartilage in osteoarthritis trials are summarized, focusing on radiography and magnetic resonance imaging (MRI). Studies having compared both imaging methodologies directly and those having established a relationship between imaging biomarkers and clinical outcomes are highlighted. To make this review of practical use, recommendations are provided as to which imaging protocols are ideal for capturing specific aspects of bone and cartilage tissue, and pitfalls in their usage are highlighted. Further, the longitudinal sensitivity to change, of different imaging methods is reported for various patient strata. From these power calculations can be accomplished, provided the strength of the treatment effect is known. In conclusion, current imaging methodologies provide powerful tools for scoring and measuring morphological and compositional aspects of most articular tissues, capturing longitudinal change with reasonable to excellent sensitivity. When employed properly, imaging has tremendous potential for ascertaining treatment effects on various joint structures, potentially over shorter time scales than required for demonstrating effects on clinical outcomes.


Exchange resonance in MDM nanolayer systems: experiment and theory.
Exchange resonance spectra of three sandwich devices containing nanolayers of Cr, Mn, Co, Ni, and Eu were recorded at 77 K. We found that these spectra are significantly different from those obtained earlier for Fe-SiO2-Au three-layer nanosandwich device. Detailed theoretical approach was developed to analyze the recorded spectra, g-factor values, and relaxation properties of the spin-polarized states in the nanolayers. We found that the g-factor values and spin-lattice relaxation rates may be adequately described by the spin-orbit scattering mechanism. Electric charge density fluctuations may also contribute to spin-lattice relaxation in nanolayers. Second-order effects in the relaxation mechanism were also briefly considered.


Interaction between diet and gastrointestinal endocrine cells.
The gastrointestinal endocrine cells are essential for life. They regulate the gastrointestinal motility, secretion, visceral sensitivity, absorption, local immune defense, cell proliferation and appetite. These cells act as sensory cells with specialized microvilli that project into the lumen that sense the gut contents (mostly nutrients and/or bacteria byproducts), and respond to luminal stimuli by releasing hormones into the lamina propria. These released hormones exert their actions by entering the circulating blood and reaching distant targets (endocrine mode), nearby structures (paracrine mode) or via afferent and efferent synaptic transmission. The mature intestinal endocrine cells are capable of expressing several hormones. A change in diet not only affects the release of gastrointestinal hormones, but also alters the densities of the gut endocrine cells. The interaction between ingested foodstuffs and the gastrointestinal endocrine cells can be utilized for the clinical management of gastrointestinal and metabolic diseases, such as irritable bowel syndrome, obesity and diabetes.


Analysis of ERCC2/XPD functional polymorphisms in systemic lupus erythematosus.
Sunlight/ultraviolet (UV) irradiation has been recognized as an important risk factor for developing systemic lupus erythematosus (SLE). However, the interpretation of genetic variations involved in UV-light sensitivity is largely unknown. Recent studies indicated that two genetic variations of ERCC2/XPD gene (rs1799793 in exon 10 and rs13181 in exon 23) have been found to exert negative influences on nucleotide excision repair system. To analyse the possible contribution of the ERCC2/XPD functional single nucleotide polymorphisms in genetic susceptibility to SLE, the rs13181 and rs1799793 SNPs in ERCC2/XPD were genotyped by polymerase chain reaction followed by restriction fragment length polymorphism analysis. Association was studied by case-control analyses using samples from 172 SLE patients and 160 healthy controls. Haplotype analysis was performed to detect the association with genetic predisposition to SLE and the clinical features. Although these two functional genetic variations are linked to several immune dysfunction-induced diseases, no statistically significant differences in allele or genotype frequencies were observed between SLE patients and controls. Haplotype analysis showed that none of ERCC2/XPD haplotypes was associated with the incidence of SLE disease, nor the preference of clinical features. In conclusion, the ERCC2/XPD functional polymorphisms analysed in this study showed no association in genetic susceptibility to SLE.


X-ray and neutron scattering measurements of ordering in a Cu46Zr54 liquid.
The structural evolution of the equilibrium and supercooled Cu46Zr54 liquids was investigated with a combination of elastic neutron scattering (with isotopic substitution) and synchrotron x-ray scattering studies. The partial pair correlation functions were determined over a wide temperature range (∼270 °C). These show that the Cu-Cu and Zr-Zr ordering increases as the temperature decreases, while the Cu-Zr ordering decreases. This surprising result is in contradiction with the results from molecular dynamics studies.


Enhanced autophagic retrograde axonal transport by dynein intermediate chain upregulation improves Aβ clearance and cognitive function in APP/PS1 double transgenic mice.
Autophagosome accumulation is observed in the distal axons of Alzheimer disease (AD) patients and AD animal models, suggesting that deficient retrograde transport and impaired autophagic clearance of beta-amyloid (A β) contribute to AD pathogenesis. Expression of the retrograde axonal transport-related protein dynein intermediate chain (DIC) is also reduced in AD patients, but the contributions of DIC to AD pathology remain elusive. This study investigated the effects of DIC expression levels on cognitive function, autophagosome axonal transport, and A β clearance in the APP/PS1 double transgenic mouse model of AD. Autophagic activity was enhanced in the hippocampus of young (3-month-old) AD mice, as evidenced by greater expression of autophagosome markers, lysosome markers, axonal transport motors (including DIC), and dynein regulatory proteins. The expression levels of autophagosome markers remained elevated, whereas those of autophagic and axonal transport proteins decreased progressively with age, accompanied by spatial learning and memory deficits, axonal autophagosome accumulation, and A β deposition. Knockdown of DIC exacerbated while overexpression improved axonal transport, autophagosome maturation, Aβ clearance, and spatial learning and memory in aged AD mice. Our study provides evidence that age-dependent failure of axonal autophagic flux contributes to AD-associated neuropathology and cognitive deficits, suggesting DIC as a potential therapeutic target for AD.


Architecting highly hydratable polymer networks to tune the water state for solar water purification.
Water purification by solar distillation is a promising technology to produce fresh water. However, solar vapor generation, is energy intensive, leading to a low water yield under natural sunlight. Therefore, developing new materials that can reduce the energy requirement of water vaporization and speed up solar water purification is highly desirable. Here, we introduce a highly hydratable light-absorbing hydrogel (h-LAH) consisting of polyvinyl alcohol and chitosan as the hydratable skeleton and polypyrrole as the light absorber, which can use less energy (<50% of bulk water) for water evaporation. We demonstrate that enhancing the hydrability of the h-LAH could change the water state and partially activate the water, hence facilitating water evaporation. The h-LAH raises the solar vapor generation to a record rate of ~3.6 kg m-2 hour-1 under 1 sun. The h-LAH-based solar still also exhibits long-term durability and antifouling functionality toward complex ionic contaminants.


Research on object-plane constraints and hologram expansion in phase retrieval algorithms for continuous-wave terahertz inline digital holography reconstruction.
In terahertz inline digital holography, zero-order diffraction light and conjugate images can cause the reconstructed image to be blurred. In this paper, three phase retrieval algorithms are applied to conduct reconstruction based on the same near-field diffraction propagation conditions and image-plane constraints. The impact of different object-plane constraints on CW terahertz inline digital holographic reconstruction is studied. The results show that in the phase retrieval algorithm it is not suitable to impose restriction on the phase when the object is not isolated in the transmission-type CW terahertz inline digital holography. In addition, the effects of zero-padding expansion, boundary replication expansion, and apodization operation on reconstructed images are studied. The results indicate that the conjugate image can be eliminated, and a better reconstructed image can be obtained by adopting an appropriate phase retrieval algorithm after the normalized hologram extending to the minimum area, which meets the applicable range of the angular spectrum reconstruction algorithm by means of boundary replication.


In vitro evaluation of a custom cutting jig and custom plate for canine tibial plateau leveling.
To design and manufacture custom titanium bone plates and a custom cutting and drill guide by use of free-form fabrication methods and to compare variables and mechanical properties of 2 canine tibial plateau leveling methods with each other and with historical control values. 10 canine tibial replicas created by rapid prototyping methods. Application time, accuracy of correction of the tibial plateau slope (TPS), presence and magnitude of rotational and angular deformation, and replica axial stiffness for 2 chevron wedge osteotomy (CWO) methods were assessed. One involved use of freehand CWO (FHCWO) and screw hole drilling, whereas the other used jig-guided CWO (JGCWO) and screw hole drilling. Replicas used for FHCWO and JGCWO methods had similar stiffness. Although JGCWO and FHCWO did not weaken the replicas, mean axial stiffness of replicas after JGCWO was higher than after FHCWO. The JGCWO method was faster than the FHCWO method. Mean +/- SD TPS after osteotomy was lower for FHCWO (4.4 +/- 1.1 degrees ) than for JGCWO (9.5 +/- 0.4 degrees ), and JGCWO was more accurate (target TPS, 8.9 degrees ). Slight varus was evident after FHCWO but not after JGCWO. Mean postoperative rotation after JGCWO and FHCWO did not differ from the target value or between methods. The JGCWO method was more accurate and more rapid and resulted in more stability than the FHCWO method. Use of custom drill guides could enhance the speed, accuracy, and stability of corrective osteotomies in dogs.


The place of cyclical therapy for the treatment of membranous nephropathy in the era of rituximab.
Primary membranous nephropathy (MN) is the most frequent cause of nephrotic syndrome in adults, due to a variety of autoantibodies, most frequently against phospholipase A2 receptor (PLA2R). In severe cases or when spontaneous remission is not achieved, immunosuppression is required. Cyclical therapy, based on glucocorticoids and cyclophosphamide on alternate months for 6 months, has proven effective to induce remission and reduce the risk of end-stage renal disease. Since the early 2000s, rituximab (RTX) has emerged as a key player in the management of MN, showing overall comparable effectiveness and likely better safety compared with the cyclical regimen, despite the lack of adequately powered trials comparing the two approaches head to head. For these reasons, RTX is now considered the agent of choice for most patients with MN. However, there are still uncertainties. Around 20-40% of patients are resistant to RTX, especially in the setting of high anti-PLA2R levels, and this drug remains relatively unexplored in patients with the most severe disease. In these scenarios, although the expanding therapeutic armamentarium is probably going to provide further options, the cyclical regimen still plays a key role as a safety net. The aim of this article is to illustrate the role of cyclical therapy in the RTX era.


POLLEN LOADS AND PROGENY VIGOR IN CUCURBITA PEPO: THE NEXT GENERATION.
Previous research on the Black Beauty bush cv. of zucchini has documented a strong positive relationship between the size of the pollen load and the vigor (performance) of the progeny. Here we report the results of three studies designed to test the hypothesis that the previously observed differences in progeny vigor are heritable. Two studies examined the transmission of the pollen load effect to subsequent generations through the ovules (female role). The third study determined if there is genetic variation for pollen performance and if the pollen load effect could be transmitted to a subsequent generation through the pollen (male role). In each of these studies the vigor of the progeny from the subsequent generation was evaluated in the greenhouse and/or the field. The results of these studies reveal (1) that the ability to sire seeds does respond to selection imposed by high pollen loads, (2) that only 23 of the 35 total traits that we measured in the three studies of transmission to subsequent generations changed in the direction predicted by the pollen competition hypothesis, (3) that only 5 of the 35 traits were significantly affected by the size of the pollen load that produced the previous generation (but all 5 were in the direction predicted by the pollen competition hypothesis), and (4) that only one study produced an overall significant difference (MANOVA) attributable to the size of the pollen load that produced the previous generation (but it too was in the direction predicted by the pollen competition hypothesis). From these experiments we conclude that pollen competition appears to play a real but minor role in the production of differences in vigor between progeny arising from low versus high pollen loads. In Black Beauty bush cv. of zucchini, maternal effects, pollen-pistil interactions, or nonrandom patterns of seed abortion must play important roles as well.


Development and structure of the glandopreputial sulcus of the human clitoris with a special reference to glandopreputial glands.
The structure and development of the sulcus between the glans and prepuce of the human clitoris have hardly been investigated. Interest in its structure was raised when in the female, in contrast to the male, glands were found to develop from the solid lamella-like precursor of the glandopreputial sulcus. It prompted a further histological analysis of the sulcus in female fetuses and newborn and an extension of that study to clitorises of adult women. The investigation showed that in the clitoris, in contrast to the penis, the transformation of the glandopreputial lamella into the open sulcus was mostly incomplete and apparently remained so throughout life. As a most striking and probably exclusively female feature, two to eight eccrine glands developed from the base of the lamella in fetuses older than 14.5 weeks gestation. These glands formed secretory coils near and occasionally inside the adjacent distal corpora cavernosa. Some glands showed atresia, cystic dilatation, and squamous metaplasia. A remarkably similar picture was observed in the adult clitorises, in which the secretory coils were often found between the large blood vessels and nerves to the glans and were connected to the sulcus by long excretory ducts. All glands revealed unmistakably eccrine features. It is suggested that their secretion moistens the female glandopreputial sulcus, which is not lubricated by urethral secretion as in the male. The findings may explain the rare clitoral phimosis, cysts, and some pilonidal sinuses.


[Postoperative complications after rectal cancer surgery and management].
Although neoadjuvant treatment such as radiotherapy, even chemoradiotherapy has already widely used in middle-low rectal cancer, surgery remains the mainstay of therapy for patients with rectal cancer. Because neoadjuvant chemoradiotherapy combined with surgery was adopted for the locally advanced rectal cancer as a standard treatment based on guidelines, postoperative complications of rectal cancer surgery caused surgeons attention and concern gradually. Some severe complications should seriously affect patient's quality of life, even patient's survival. Therefore, surgeons should be fully aware of the severity of complications, and try to avoid the occurrence of complications. Thoroughly knowing the significance of comprehensive treatment in rectal cancer, meanwhile, realizing associated complications following rectal cancer operation, and choosing reasonable, standard, effective and economic management will result in obvious increase of therapeutic level in rectal cancer surgery in China. Through the literature review combined with our clinical experience, this paper analyzes the reasons of seven associated complications, including intra-operational bleeding, bladder damage, postoperative anastomotic leakage, rectovaginal fistula, postoperative bleeding, postoperative ileus, postoperative anastomotic stenosis and anterior resection syndrome, and shares the managements in order to provide more references to surgeons.


Heterogeneity of memory B cells.
Potential solid organ transplant recipients broadly sensitized to HLA have long wait times, low transplant rates and poor outcomes. The new kidney allocation system has improved access to the most highly sensitized recipients; however, their long-term outcomes are unknown. Emerging data suggest that memory B cell repertoire is broader than the plasma cell repertoire, therefore, despite refinements in anti-HLA antibody detection technology, donor-specific HLA- specific memory B cells may in fact be present in some, if not most, highly sensitized recipients with no detectable donor-specific antibodies. In addition, new findings have underscored the heterogeneity in memory B cell generation, and in the signals that determine memory versus plasma cell fate during primary antigen encounter, as well as memory B cell differentiation upon antigen reencounter into plasma cells or reentry into germinal centers to subsequently emerge as higher affinity and class-switched plasma cells. Thus, heterogeneity memory B cells generation may affect the efficacy of specific immunomodulation during the recall response. We propose that the ability to quantify donor-specific B cell in transplant recipients is urgently required to provide insights into the mechanisms of sensitization and recall, and for the early detection of acute and chronic AMR.


Distribution of HPV Genotypes in Patients with a Diagnosis of Anal Cancer in an Italian Region.
Objectives: Anal cancer is a rare disease. However, its incidence is increasing in some population groups. Infection caused by Human Papillomavirus (HPV) is strongly associated with the risk of anal cancer, whose variability depends on samples, histology, and HPV detection methods. The aim of the study was to assess prevalence and distribution of HPV genotypes in patients diagnosed with anal carcinoma. Methods: An observational, retrospective study was carried out in a tertiary care hospital in North Sardinia, Italy. Specimens of anal cancer diagnosed from 2002-2018 were selected. Demographic, epidemiological, and clinical variables were collected to assess their relationship with the occurrence of anal cancer. Results: The overall HPV positivity was 70.0% (21/30), with HPV-16 being the predominant genotype (~85%). The highest prevalence of anal cancer was in patients aged ≥55 years. HPV positivity was higher in women (p-value > 0.05) and in moderately differentiated samples (G2) (p-value < 0.05). p16INK4a and E6-transcript positivity were found in 57% and 24% of the HPV positive samples, respectively. The OS (overall survival) showed a not statistically significant difference in prognosis between HPV positive sand negatives (10, 47.6%, vs. 4, 44.4%; p-value = 0.25). Conclusions: HPV-DNA and p16INK4a positivity confirmed the role of HPV in anal carcinoma. Our findings could support the implementation and scale-up of HPV vaccination in males and females to decrease the incidence of HPV-associated cancers. Further studies are needed to better clarify the prognostic role of HPV/p16 status.


Unscrambling the Influence of Sodium Cation on the Structure, Bioactivity, and Erythrocyte Compatibility of 45S5 Bioactive Glass.
The 45S5 bioglass uttering Class A bioactivity promotes both osteoconduction as well as osteoinduction. Though one of the higher reactive bioactive materials known with structural and physiological influence upon ionic modulation, poor mechanical properties are perceived. The possible solution to overcome the weak stability is to choose material's composition that provides retained bioactivity and improved mechanical stability. Meanwhile, primary burst out of Na+ ions increases the local pH, harms cell life, and acts as a well-known disruptive modifying species that weakens the bioactive glass network, decreasing network connectivity, showing faster degradation and lowering mechanical stability. Therefore, in this study, more detailed systematic exploration on structural influence of sodium monovalent cation and its behavior on physiological environment was genuinely studied and reported that bioactivity of the bioactive glass can be highly achieved even without Na+ ions. The result exhibits benefits of sodium free bioactive glass (denoted as No Na+ BG) over Na+ BG and exhibits improved mechanical stability and also possible degradability, having in-built apatite phase even before immersion in simulated body fluid (SBF). Also, sodium free bioglass proved as a superior candidate for erythrocyte compatibility with rapid clotting tendency on interaction with blood and a promising replacement for 45S5 bioglass in all aspects especially in mechanical stability view, which can withstand more than 5 months in phosphate buffer saline (PBS).


Reversal of spherical remodelling of the left atrium after pulmonary vein isolation: incidence and predictors.
Pulmonary vein isolation (PVI) induces left atrial (LA) volume reduction, known as reverse remodelling (RR). The related changes in LA shape have not yet been evaluated. Left atrial sphericity (LASP) is a new shape-based marker of remodelling that compares LA geometry and a perfect sphere and is a powerful predictor of PVI success. We aimed to evaluate the effect of PVI on LASP and describe the concept of spherical and volumetric RR. Left atrial sphericity and volume were automatically obtained with self-customized software using a magnetic resonance imaging-based three-dimensional reconstruction of LA. Reverse remodelling was defined as improvement in LASP (spherical RR) or volume reduction (volumetric RR). In a series of 102 patients, spherical and volumetric RR was observed in 42.2 and 69%, respectively. Patients with paroxysmal atrial fibrillation (AF) had higher probability to present spherical RR as compared with patients with persistent AF (50.8 vs. 29.3%, P = 0.03). Patients with persistent AF showed significant post-procedural worsening of LASP (81.9 vs. 82.9%, P = 0.04). Patients with no recurrence showed a trend towards a higher proportion of spherical RR compared with those with recurrences (46.2 vs. 32.4%, respectively); no differences were observed in volumetric RR (62.1 vs. 62.9%, respectively). Paroxysmal AF was the only independent predictor of spherical RR. Pulmonary vein isolation leads to spherical RR in a substantial proportion of patients, and in higher proportion of patients with paroxysmal AF. Reverse remodelling may be caused by a combination of scarring and myocardial structural recovery. Changes in LASP might be more specific than volume reduction to detect favourable remodelling.


Effects of dietary protein content on renal parameters in normal cats.
This study evaluates the effect of dietary protein content on renal parameters in 23 healthy spayed female cats. The objective was to determine if cats eating diets high in protein will have higher serum urea nitrogen (UN) and creatinine values without a detectable change in kidney function, as assessed by urinalysis. A single random cross-over design was used. Cats were fed a standard maintenance diet for at least 1 month prior to the dietary trial. They were fed in two phases. For the first phase, cats were randomly assigned to receive either a high protein [HP=46% metabolizable energy (ME)] or low protein (LP=26% ME) diet. For the second phase, cats were fed whichever diet they were not fed during the phase I period. Blood and urine samples were collected at 2-week intervals for the duration of the study (10 weeks). UN, albumin, alanine aminotransferase and urine specific gravity were significantly higher, and creatinine and phosphorus were significantly lower (P<0.05) when cats were fed the HP diet as compared to when they were fed the LP diet, although none of the mean values were found to be outside of the corresponding reference interval. Dietary intake can result in clinically significant changes in UN and statistically significantly changes in several other biochemical analytes, although all analytes are likely to remain within normal reference intervals. Therefore, an accurate dietary history is necessary to help determine if renal parameters are being influenced by diet in a particular patient.


Optimal window of caloric restriction onset limits its beneficial impact on T-cell senescence in primates.
We have recently shown in non-human primates that caloric restriction (CR) initiated during adulthood can delay T-cell aging and preserve naïve CD8 and CD4 T cells into advanced age. An important question is whether CR can be initiated at any time in life, and whether age at the time of onset would modulate the beneficial effects of CR. In the current study, we evaluated the impact of CR started before puberty or during advanced age on T-cell senescence and compared it to the effects of CR started in early adulthood. Our data demonstrate that the beneficial effects of adult-onset CR on T-cell aging were lost by both early and late CR onset. In fact, some of our results suggest that inappropriate initiation of CR may be harmful to the maintenance of T-cell function. This suggests that there may be an optimal window during adulthood where CR can delay immune senescence and improve correlates of immunity in primates.


A hybrid CPU-GPGPU approach for real-time elastography.
Ultrasound elastography is becoming a widely available clinical imaging tool. In recent years, several real- time elastography algorithms have been proposed; however, most of these algorithms achieve real-time frame rates through compromises in elastographic image quality. Cross-correlation- based elastographic techniques are known to provide high- quality elastographic estimates, but they are computationally intense and usually not suitable for real-time clinical applications. Recently, the use of massively parallel general purpose graphics processing units (GPGPUs) for accelerating computationally intense operations in biomedical applications has received great interest. In this study, we investigate the use of the GPGPU to speed up generation of cross-correlation-based elastograms and achieve real-time frame rates while preserving elastographic image quality. We propose and statistically analyze performance of a new hybrid model of computation suitable for elastography applications in which sequential code is executed on the CPU and parallel code is executed on the GPGPU. Our results indicate that the proposed hybrid approach yields optimal results and adequately addresses the trade-off between speed and quality.


Magnetic ordering in the rare-Earth hexaborides.
Magnetic ordering in the rare-earth hexaborides has been studied by a variety of methods, and a clear correlation with the presence of conduction electrons has been found.


Influenza vaccination: options and issues.
Currently available vaccines have similar efficacy if they are matched to the most prevalent circulating strains. They also have comparable adverse effect profiles. The choice of a specific preparation of vaccine therefore requires consideration of cost, purity of the vaccine preparation in terms of the amount of egg protein and endotoxin, allergy to different constituents of the vaccine, reactogenicity profiles, as well as the preferred route of administration. Intradermal injection of the vaccine appears to be a viable alternative to the traditional intramuscular administration with the additional benefit of requiring a smaller volume of vaccine. Despite the documented benefits in various community and institutional settings, influenza vaccination has been underutilised by most target groups. A major obstacle to broader coverage of vaccination is the perceived ineffectiveness of the vaccine and the relatively benign nature of the illness in most patients. Uptake of vaccine among target populations, especially health care workers, needs to be improved through a concerted effort between frontline clinicians and health authorities.


NRAS Status Determines Sensitivity to SHP2 Inhibitor Combination Therapies Targeting the RAS-MAPK Pathway in Neuroblastoma.
Survival for high-risk neuroblastoma remains poor and treatment for relapsed disease rarely leads to long-term cures. Large sequencing studies of neuroblastoma tumors from diagnosis have not identified common targetable driver mutations other than the 10% of tumors that harbor mutations in the anaplastic lymphoma kinase (ALK) gene. However, at neuroblastoma recurrence, more frequent mutations in genes in the RAS-MAPK pathway have been detected. The PTPN11-encoded tyrosine phosphatase SHP2 is an activator of the RAS pathway, and we and others have shown that pharmacologic inhibition of SHP2 suppresses the growth of various tumor types harboring KRAS mutations such as pancreatic and lung cancers. Here we report inhibition of growth and downstream RAS-MAPK signaling in neuroblastoma cells in response to treatment with the SHP2 inhibitors SHP099, II-B08, and RMC-4550. However, neuroblastoma cell lines harboring endogenous NRAS Q61K mutation (which is commonly detected at relapse) or isogenic neuroblastoma cells engineered to overexpress NRASQ61K were distinctly resistant to SHP2 inhibitors. Combinations of SHP2 inhibitors with other RAS pathway inhibitors such as trametinib, vemurafenib, and ulixertinib were synergistic and reversed resistance to SHP2 inhibition in neuroblastoma in vitro and in vivo. These results suggest for the first time that combination therapies targeting SHP2 and other components of the RAS-MAPK pathway may be effective against conventional therapy-resistant relapsed neuroblastoma, including those that have acquired NRAS mutations. SIGNIFICANCE: These findings suggest that conventional therapy-resistant, relapsed neuroblastoma may be effectively treated via combined inhibition of SHP2 and MEK or ERK of the RAS-MAPK pathway.


Transapical transcatheter aortic valve implantation in a heart transplant recipient with severely depressed left ventricular function.
Transcatheter aortic valve implantation (TAVI) is becoming a valuable alternative to surgical aortic valve replacement in non-operable and high-risk surgical patients. As the population of heart donors and recipients ages, the prevalence of degenerative valvular disease after transplantation will increase. The optimal treatment strategy of valvulopathies in these patients with extensive comorbidity is still unknown because of insufficient published experience. We present a heart transplant recipient with renal failure, systolic heart failure and severe aortic stenosis who was successfully treated with transapical TAVI.


The effect of mouth closure on drooling and speech.
The efficacy of a feeding program to decrease drooling and increase vocalizations by promoting mouth closure was explored in two boys with cerebral palsy through the use of oral facilitation techniques. A single-subject ABA design was repeated across subjects. Baseline 1 consisted of 5 (Subject 1) and 10 (Subject 2) half-hour sessions during which the subjects were fed applesauce and apple juice without treatment. A 1-hr play session followed, during which saliva was collected on an absorbent bib and weighted. Bilabial vocalizations produced by the subjects were recorded and counted. A 12-session intervention phase followed, which was identical to the baseline phase except that treatment was implemented. Treatment consisted of techniques to reduce tactile hypersensitivity before feeding and the application of jaw control during feeding. Baseline 2 consisted of 7 sessions identical to baseline 1. The results suggest that this intervention decreases drooling but does not increase bilabial vocalizations. Longer treatment and follow-up may be needed in future efficacy studies.


A Clinical Decision Support System for Increasing Compliance with Protocols in Chemotherapy of Children with Acute Lymphoblastic Leukemia.
In this survey, a protocol-based Chemotherapy Prescription Decision Support System (CPDSS) was designed and evaluated to reduce medication errors in the chemotherapy process of children with ALL. The CPDSS algorithm was extracted by the software development team based on the protocol used by doctors to treat children with ALL. The ASP.Net MVC and SQL Server 2016 programming languages were used to develop the system. A 3-step evaluation (technical, retrospective, and user satisfaction) was performed on CPDSS designed at 2 children's hospitals in Tehran. The data were analyzed using descriptive statistics. At the technical evaluation step, users provided recommendations included in the system. In the retrospective CPDSS evaluation step, 1281 prescribed doses of the drugs related to 30 patients were entered into the system. CPDSS detected 735 cases of protocol deviations and 57 (95%, CI = 1.25-2.55) errors in prescribed chemotherapy for children with ALL. In the user satisfaction evaluation, the users approved two dimensions of the user interface and functionality of the system. With the provision of alerts, the CPDSS can help increase compliance with chemotherapy protocols and decrease the chemotherapy prescribing errors that can improve patient safety.


Disruption of tight junctions during traversal of the respiratory epithelium by Burkholderia cenocepacia.
Burkholderia cenocepacia is an opportunistic bacterial species capable of causing life-threatening respiratory tract infection in persons with cystic fibrosis (CF). Unlike most other pathogens in CF, which typically remain confined to the endobronchial spaces, B. cenocepacia can traverse airway epithelium to cause bacteremia and sepsis. The mechanisms by which this occurs, however, are unknown. We examined the transmigration of B. cenocepacia through polarized respiratory epithelium. Representatives of three "epidemic" lineages common among CF patients in North America were able to traverse polarized 16HBE14o- cells in vitro. Transmigration of bacteria was associated with significant perturbations in epithelial permeability, as measured by a loss of transepithelial electrical resistance and increased flux of bovine serum albumin across the cell layer. Terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling and trypan blue exclusion assays, as well as lactate dehydrogenase levels, did not indicate excessive cytotoxicity or cell death in infected cell layers. Rather, confocal fluorescence microscopy demonstrated the loss of occludin from tight junctions. In contrast, zonula occludens 1 was well preserved along intercellular borders. Western blot analysis showed a shift in the major occludin isoforms from high- to low-phosphorylation states during infection. These observations suggest that B. cenocepacia traverses polarized respiratory epithelium by the dephosphorylation and dissociation of occludin from the tight-junction complex.


Experimental study of teaching critical thinking in civic education in Taiwanese junior high school.
To effectively respond to the need for greater CT (critical thinking) in the classroom, this study examines the effects of cultivating CT skills within civic education to maximize its potential. Despite realizing the importance of CT in education, schools do not tend to apply it. Furthermore, since students frequently do not raise questions or otherwise think critically, CT modules are incorporated into civic learning to encourage students to question facts, interpret and analyse evidence, make reasoned inferences regarding events, and independently develop informed opinions. These skills are fundamental to cultivating an informed and humane citizenry. This investigation examines how teaching critical thinking in civic education affects the CT skills and disposition of junior high school students. The participants were two classes of eighth grade students in southern Taiwan, and were distributed into experimental and control groups. Each group comprised 34 students, with the experimental group containing 16 boys and 18 girls and the control group containing 18 boys and 16 girls. The teaching experiment in this study was initiated and performed using equivalent pre-test and post-test group design. During the 10-week experiment, the experimental group was taught using CT instruction in civic education, while the control group was not taught with any CT programme. The data analysis included: quantitative statistics of the two main sets of instruments for measuring the CT ability and disposition of experimental treatment, qualitative analysis of learning sheets, and surveys of student perceptions of instruction in CT. Statistical analysis showed that the experimental group significantly outperformed the control group on the scales of CT skill and disposition, but did not perform well on certain subscales (e.g. recognition of assumptions and deductions, truth-seeking, analyticity, and inquisitiveness). Notably however, when triangulating with qualitative analysis of learner transcripts and surveys, learners attempted to perform various dimensions of CT skill and disposition to various degrees. Student surveys indicated that the CT programme fostered their active listening and respect for different ideas, and moreover they learned to tolerate divergent views and examine their ideas for possible bias. The programme boosted student learning interest and sense of accomplishment, and nurtured their teamwork/communicative skills. Furthermore, a few students experienced improvements in their speaking skills and courage as their confidence and ability to express themselves improved. Finally, limitations of this investigation and implications for further research are discussed.


Surface hardness and wear of glass ionomers and compomers.
To determine the surface hardness and in vitro wear of glass ionomer cements (GIC) and compomers in comparison with a resin composite and to test for a possible correlation between the determined hardness and wear values. Samples were made of conventional GIC (Ketac-Fil, Fuji II, Fuji IX, Ketac-Prototype = Ketac-Molar), resin-modified GIC (Photac-Fil, Fuji II LC, Vitremer), compomers (Dyract, Compoglass) and resin composite (Z100) for the hardness test (n = 10) and for the wear test (n = 3). The materials were handled according to the manufacturers' instructions. The samples were stored in deionized water for 48 hours at 37 degrees C prior to the determination of Rockwell hardness and three-body in vitro wear. Hardness could not be measured on Ketac-Fil and Fuji II as the samples fractured before a reading was obtainable. Statistically significant differences were found in hardness and wear between brands and between the four types of material. Surface hardness: resin-modified GIC < conventional GIC = compomers < resin composite. In vitro wear: resin-modified GIC > compomers > conventional GIC > resin composite. A negative correlation between hardness and wear was recorded.