The therapeutic alliance in treatment of traumatized youths: relation to outcome in a randomized clinical trial.
We examined the contribution of alliance to the outcome of therapy with traumatized youths across two different treatment conditions (trauma-focused cognitive behavioral therapy [TF-CBT] and therapy as usual [TAU]). Participants were 156 youths (M age = 15.1 years, range = 10-18; 79.5% girls), randomly assigned to TF-CBT or TAU. Symptoms were assessed pretreatment, midtreatment (Session 6), and posttreatment (Session 15). Alliance was assessed after Sessions 1 and 6, using the Therapeutic Alliance Scale for Children-Revised (TASC-R). Alliance scores were comparable across treatment conditions, but TF-CBT participants had significantly lower posttraumatic stress symptoms (PTSS) posttreatment (d = 0.51). Hierarchical regression analyses showed that there were no significant alliance effects in models without an Alliance × Treatment Group interaction: Alliance ratings were significant predictors of reduction in PTSS (Est. = -0.53, p = .003, 95% confidence interval [CI] = -0.87 to -0.18) and additional outcomes measured in TF-CBT but not in TAU (PTSS posttreatment: Est. = 0.01, p = .647, 95% CI = -0.29 to 0.47). This study was the first to investigate the contribution of alliance to outcome among adolescents with posttraumatic symptoms, treated with TF-CBT or TAU. Our findings indicated that there was an important interaction between alliance and therapeutic approach, as alliance predicted outcome in TF-CBT, but not in the nonspecific treatment condition. A positive working relationship appeared to be especially important in the context of this evidence-based treatment, which requires youth involvement in specific therapy tasks. Further, findings showed that use of a manual did not compromise alliance formation.


Dynamic object perception by pigeons: discrimination of action in video presentations.
Two experiments examined the discrimination by pigeons of relative motion using computer-generated video stimuli. Using a go/no-go procedure, pigeons were tested with video stimuli in which the camera's perspective went either "around" or "through" an approaching object in a semi-realistic context. Experiment 1 found that pigeons could learn this discrimination and transfer it to videos composed from novel objects. Experiment 2 found that the order of the video's frames was critical to the discrimination of the videos. We hypothesize that the pigeons perceived a three-dimensional representation of the objects and the camera's relative motion and used this as the primary basis for discrimination. It is proposed that the pigeons might be able to form generalized natural categories for the different kinds of motions portrayed in the videos.


Wrinkles are neither beautiful nor nice: The effect of facial wrinkles on person perception and interpersonal closeness.
With age, we develop facial wrinkles, which change the appearance of the face making it less attractive. Winkles and folds also "mimic" facial expressions, such that older neutral faces appear more emotional. Both processes can influence first impressions negatively. We obtained ratings of attractiveness, closeness, and emotional expressivity as well as first impressions from 353 participants, recruited from Prolific, for avatars with and without facial wrinkles. Older appearing, wrinkled faces were judged as less attractive and less favorably on traits related to pleasantness and trustworthiness, they were perceived as showing more negative emotions and participants rated themselves as less close. The effects on first impressions and perceived closeness were mediated by attractiveness and perceived negative emotions. These findings suggest that in initial encounters older people may often be perceived as less pleasant for no other reason than the wrinkles in their face and the judgments of attractiveness and emotionality associated with them.


Anatomical relationships of intracavernous internal carotid artery to intracavernous neural structures.
The objective is to correlate the intracavernous internal carotid artery (ICA) with the position of the intracavernous neural structures. The cavernous sinuses of nine injected cadaveric heads were dissected bilaterally. As measured on computed tomographic angiograms from 100 adults, anatomical relationships and measurements of intracavernous ICA and neural structures were studied and correlated to the intracavernous ICA curvature. Intracavernous ICAs were classified as normal and redundant. The meningohypophyseal trunk (MHT) of normal ICAs appeared to be closely related to the abducens nerve compared with redundant ICAs (5.5 ± 2.1 mm versus 10.0 ± 2.5 mm, respectively; p = 0.001). The position of the inferolateral trunk (ILT) varied along the horizontal segment of the intracavernous ICA. On imaging studies the ICA curvature correlated with the kyphotic degree of the skull and similarity of the ICA curvature between sides. The safety margin for preventing iatrogenic intracavernous nerve injury during surgical exploration or transarterial embolization of vascular lesions around the MHT is high with redundant ICAs. In contrast, a transvenous endovascular approach via the inferior petrosal sinus may be too distant to reach the MHT when ICAs are redundant. Approaching lesions of the inferolateral trunk may be the same regardless of ICA type.


Comparative histometric analysis of the effects of high-intensity focused ultrasound and radiofrequency on skin.
High-intensity focused ultrasound (HIFU) and radiofrequency (RF) are used for non-invasive skin tightening. Neocollagenesis and neoelastogenesis have been reported to have a mechanism of controlled thermal injury. To compare neocollagenesis and neoelastogenesis in each layer of the dermis after each session of HIFU and monopolar RF. We analyzed the area fraction of collagen and elastic fibers using the Masson's Trichrome and Victoria blue special stains, respectively, before and after 2 months of treatments. Histometric analyses were performed in each layer of the dermis, including the papillary dermis, and upper, mid, and deep reticular dermis. Monopolar RF led to neocollagenesis in the papillary dermis, and upper, mid, and deep reticular dermis, and neoelastogenesis in the papillary dermis, and upper and mid reticular dermis. HIFU led to neocollagenesis in the mid and deep reticular dermis and neoelastogenesis in the deep reticular dermis. Among these treatment methods, HIFU showed the highest level of neocollagenesis and neoelastogenesis in the deep reticular dermis. HIFU affects deep tissues and impacts focal regions. Monopolar RF also affects deep tissues, but impacts diffuse regions. We believe these data provide further insight into effective skin tightening.


Antioxidant effect of Aloe vera gel extract in streptozotocin-induced diabetes in rats.
In the present study, an attempt has been made to evaluate the presence of antioxidant property in the alcoholic extract of Aloe vera leaf gel. Oral administration of Aloe vera gel extract at a concentration of 300 mg/kg to diabetic rats significantly decreased the levels of blood glucose, glycosylated hemoglobin and increased hemoglobin. The increased levels of lipid peroxidation and hydroperoxides in tissues of diabetic rats were reverted back to near normal levels after the treatment with gel extract. The extract treatment also resulted in a significant increase in reduced glutathione, superoxide dismutase, catalase, glutathione peroxidase and glutathione-S-transferase in the liver and kidney of diabetic rats. These results clearly show the antioxidant property of Aloe vera gel extract. The extract was also more effective than glibenclamide in restoring the values of these parameters.


Temporal development of near-native functional properties and correlations with qMRI in self-assembling fibrocartilage treated with exogenous lysyl oxidase homolog 2.
Advances in cartilage tissue engineering have led to constructs with mechanical integrity and biochemical composition increasingly resembling that of native tissues. In particular, collagen cross-linking with lysyl oxidase has been used to significantly enhance the mechanical properties of engineered neotissues. In this study, development of collagen cross-links over time, and correlations with tensile properties, were examined in self-assembling neotissues. Additionally, quantitative MRI metrics were examined in relation to construct mechanical properties as well as pyridinoline cross-link content and other engineered tissue components. Scaffold-free meniscus fibrocartilage was cultured in the presence of exogenous lysyl oxidase, and assessed at multiple time points over 8weeks starting from the first week of culture. Engineered constructs demonstrated a 9.9-fold increase in pyridinoline content, reaching 77% of native tissue values, after 8weeks of culture. Additionally, engineered tissues reached 66% of the Young's modulus in the radial direction of native tissues. Further, collagen cross-links were found to correlate with tensile properties, contributing 67% of the tensile strength of engineered neocartilages. Finally, examination of quantitative MRI metrics revealed several correlations with mechanical and biochemical properties of engineered constructs. This study displays the importance of culture duration for collagen cross-link formation, and demonstrates the potential of quantitative MRI in investigating properties of engineered cartilages. This is the first study to demonstrate near-native cross-link content in an engineered tissue, and the first study to quantify pyridinoline cross-link development over time in a self-assembling tissue. Additionally, this work shows the relative contributions of collagen and pyridinoline to the tensile properties of collagenous tissue for the first time. Furthermore, this is the first investigation to identify a relationship between qMRI metrics and the pyridinoline cross-link content of an engineered collagenous tissue.


Deletion of Transferrin Receptor 1 in Parvalbumin Interneurons Induces a Hereditary Spastic Paraplegia-Like Phenotype.
Hereditary spastic paraplegia (HSP) is a severe neurodegenerative movement disorder, the underlying pathophysiology of which remains poorly understood. Mounting evidence has suggested that iron homeostasis dysregulation can lead to motor function impairment. However, whether deficits in iron homeostasis are involved in the pathophysiology of HSP remains unknown. To address this knowledge gap, we focused on parvalbumin-positive (PV+) interneurons, a large category of inhibitory neurons in the central nervous system, which play a critical role in motor regulation. The PV+ interneuron-specific deletion of the gene encoding transferrin receptor 1 (TFR1), a key component of the neuronal iron uptake machinery, induced severe progressive motor deficits in both male and female mice. In addition, we observed skeletal muscle atrophy, axon degeneration in the spinal cord dorsal column, and alterations in the expression of HSP-related proteins in male mice with Tfr1 deletion in the PV+ interneurons. These phenotypes were highly consistent with the core clinical features of HSP cases. Furthermore, the effects on motor function induced by Tfr1 ablation in PV+ interneurons were mostly concentrated in the dorsal spinal cord; however, iron repletion partly rescued the motor defects and axon loss seen in both sexes of conditional Tfr1 mutant mice. Our study describes a new mouse model for mechanistic and therapeutic studies relating to HSP and provides novel insights into iron metabolism in spinal cord PV+ interneurons and its role in the regulation of motor functions.SIGNIFICANCE STATEMENT Iron is crucial for neuronal functioning. Mounting evidence suggests that iron homeostasis dysregulation can induce motor function deficits. Transferrin receptor 1 (TFR1) is thought to be the key component in neuronal iron uptake. We found that deletion of Tfr1 in parvalbumin-positive (PV+) interneurons in mice induced severe progressive motor deficits, skeletal muscle atrophy, axon degeneration in the spinal cord dorsal column, and alterations in the expression of hereditary spastic paraplegia (HSP)-related proteins. These phenotypes were highly consistent with the core clinical features of HSP cases and partly rescued by iron repletion. This study describes a new mouse model for the study of HSP and provides novel insights into iron metabolism in spinal cord PV+ interneurons.


Effect of early mobilization on healing of nerve repair: histologic observations in a canine model.
The effect of early mobilization on the healing of nerve repair was studied in a canine model. Median and ulnar nerves in the left wrist of 16 adult mongrel dogs were transected and immediately repaired. No motion of the repaired forelimb was allowed in the immobilized group (n = 10), while controlled passive motion between 30 and 90 degrees of wrist flexion was begun on the first postoperative day for 10 minutes twice daily in the mobilized group (n = 6). The pattern of revascularization and collagen formation at neurorrhaphy was examined by transillumination of India ink-injected specimen and by conventional histologic sections. Revascularization of nerve repair was found to occur by ingrowth of capillaries from proximal and distal nerve ends, which typically crossed the neurorrhaphy by 3 weeks in the immobilized group. Following early mobilization, there was a persistent "hypovascular zone" at the nerve repair site for up to 6 weeks. In addition, more scar tissue was generated by early motion according to gross observation and quantitative collagen analysis. Early mobilization, therefore, seems to impede nerve regeneration by delaying revascularization and enhancing scar formation.


Fusion rates in multilevel, instrumented anterior cervical fusion for degenerative disease with and without the use of bone morphogenetic protein.
The goal of this study was to compare the rates of solid arthrodesis and complications following multilevel, instrumented anterior cervical fusion in patients treated with and without bone morphogenetic protein (BMP). The authors conducted a retrospective cohort study of patients who underwent multilevel (2+ level) anterior cervical fusions performed for degenerative disc disease with or without the concurrent use of BMP-2 from 1997 to 2012. The dosage throughout the study ranged from 2.1 to 0.26 mg/level (mean 1.0 mg/level). All patients were evaluated postoperatively by means of radiographs and CT scans to determine fusion status. The overall fusion rate for the patients treated without BMP (n = 23) was 82.6% compared with a 100% fusion rate in the group treated with BMP (n = 22) (p = 0.04). The pseudarthrosis rates increased with number of fusion levels in patients who did not receive BMP, whereas all patients in the group treated with BMP had solid arthrodesis. Furthermore, there were 2 instrumentation failures in the non-BMP group. There was a direct correlation between the incidence of complications and the dosage of BMP used per level, with no complications reported at doses equal to or less than 1.1 mg/level. The overall rate of bony arthrodesis was increased following the use of BMP in multilevel anterior cervical fusion. Traditional methods without BMP had a high rate of pseudarthrosis. The complications associated with the use of BMP appeared to be dose related and of low incidence when BMP is used in doses equal to or less than 1.1 mg/level.


[Extracorporeal shock-wave lithotripsy as treatment of lithiasis in horseshoe kidney].
Horseshoe kidney is the most frequent renal congenital anomaly, with an estimated general population prevalence of 1/400-500. This entity may be asymptomatic for the entire lifetime, or produce symptoms from associated complications such as lithiasis, hydronephrosis, or recurrent infections. The incidence of lithiasis varies between 20% and 80% of the patients. The treatment of lithiasis in this entity is controversial due to problems derived from the anatomy of the kidney and its drainage when stones are treated with external shock wave lithotripsy (ESWL). We tried to evaluate the efficacy of ESWL treatment and to establish the ideal conditions in which this technique may be considered treatment of choice. A total of 25 patients were evaluated (17 males and 8 females). Patient age ranged from 8 to 75 year old. Three patients presented with bilateral lithiasis. 27 renal units were treated, 18 of which were left (66.7%) and 7 right. Stone size was measured in cm2 of surface. Lithiasis was located at the renal pelvis in 13 cases (48.1%), and lower calyx in 7 (25.9%). Four patients required JJ stent insertion. Mean number of shock waves per session was 3480 (range 1000-4000). Two Shock wave generators were used for treatment, the Dornier lithotripter S and the Dornier MFL-5000, with a range of 10-120 KV for the first one and 14-23 KV for the second. Follow up KUB X-rays were performed at 3, 6 and 12 months to evaluate fragmentation and elimination rates. In our series total fragmentation was 85.2%, and partial fragmentation (fragments > 6 mm) 14.8%. Elimination rates were satisfactory with 37.4% total elimination, and 48.1% partial elimination (Fragments < 6 mm). 14.8% of the patients had no elimination at all. We consider ESWL the first therapeutic option for cases of lithiasis with a mean area 4 cm2 or less and pelvic location. In case of great lithiasic areas other therapeutic options should be considered (open surgery or percutaneous nephrolithotomy) either in monotherapy or complementary to ESWL.


Ly6G ligation blocks recruitment of neutrophils via a β2-integrin-dependent mechanism.
Ly6G is a glycosylphosphatidylinositol (GPI)-anchored protein of unknown function that is commonly targeted to induce experimental neutrophil depletion in mice. In the present study, we found that doses of anti-Ly6G Abs too low to produce sustained neutropenia remained capable of inhibiting experimental arthritis, leaving joint tissues free of infiltrating neutrophils. Thioglycollate-stimulated peritonitis was also attenuated. No alteration in neutrophil apoptosis was observed, implicating impaired recruitment. Indeed, Ly6G ligation abrogated neutrophil migration toward LTB(4) and other chemoattractants in a transwell system. Exploring the basis for this blockade, we identified colocalization of Ly6G and β2-integrins by confocal microscopy and confirmed close association by both coimmunoprecipitation and fluorescence lifetime imaging microscopy. Anti-Ly6G Ab impaired surface expression of β2-integrins in LTB(4)-stimulated neutrophils and mimicked CD11a blockade in inhibiting both ICAM-1 binding and firm adhesion to activated endothelium under flow conditions. Correspondingly, migration of β2-integrin-deficient neutrophils was no longer inhibited by anti-Ly6G. These results demonstrate that experimental targeting of Ly6G has functional effects on the neutrophil population and identify a previously unappreciated role for Ly6G as a modulator of neutrophil migration to sites of inflammation via a β2-integrin-dependent mechanism.


Genetic risk stratification to reduce inhibitor development in the early treatment of hemophilia A: a SIPPET analysis.
A recent randomized trial, the Survey of Inhibitors in Plasma-Product Exposed Toddlers (SIPPET), showed a higher risk of inhibitor development with recombinant factor VIII (rFVIII) than plasma-derived concentrates (pdFVIII). We investigated whether risk stratification by F8 mutation identifies patients who do not suffer this deleterious effect of rFVIII. Among 235 randomized patients with severe hemophilia A previously untreated with FVIII concentrate, 197 with null mutations were classified as high risk and 38 with non-null mutations were classified as low risk. With pdFVIII, no inhibitors occurred in those with low genetic risk, whereas high-risk patients had a cumulative incidence of 31%. The risk among low- and high-risk patients did not differ much when they were treated with rFVIII (43% and 47%, respectively). This implies that patients with low genetic risk suffer disproportionate harm when treated with rFVIII (risk increment 43%), as also shown by the number needed to harm with rFVIII, which was 6.3 for genetically high-risk patients and only 2.3 for low-risk patients. Risk stratification by F8 mutation does not identify patients who can be safely treated with rFVIII, as relates to immunogenicity. This trial was registered at the European Clinical Trials Database (EudraCT) as #2009-011186-88 and at www.clinicaltrials.gov as #NCT01064284.


Detection of emerging contaminants (UV filters, UV stabilizers and musks) in marine mussels from Portuguese coast by QuEChERS extraction and GC-MS/MS.
The UV filters and musk fragrances have come into focus because these compounds are contained and increasingly used not only in sunscreen products but also in many products of daily use, such as cosmetics, skin creams, plastics or varnish. In view of this, the main objective of the present work was to develop and validate a method for the determination of three UV filters, two UV stabilizers and four musks in mussel samples (Mytilus galloprovincialis). The procedure combined a QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) extraction method with an analysis by gas chromatography-tandem mass spectrometry (GC-MS/MS). The methodology allowed the determination of target analytes at trace concentration levels (ng/g), with mean recoveries ranging from 91 to 112%. A monitoring study was conducted in four beaches in the Portuguese coast which are impacted by recreational activities and outflow of treated waste water effluents in rivers. The results are used to assess the occurrence of UV filters in comparison with UV stabilizers and musk fragrances which indicate other activities than bathing.


Significance of sciatic scoliotic list in operated patients with lumbar disc herniation.
The authors retrospectively reviewed the relation between the location of disc herniation and pre- and postoperative changes in sciatic scoliotic list in 40 patients with surgically confirmed lumbar disc herniation who had sciatic scoliotic list with postoperative recovery. Clinical factors associated with scoliosis also were included. To evaluate the significance and pathomechanism of sciatic scoliotic list. The proposed causes of lumbar sciatic scoliosis mainly imply an alleviation of nerve root irritation in relation to the anatomic location of disc herniation relative to the nerve root. The pre- and postoperative serial Cobb angle between L1 and L5 in anteroposterior lumbar radiographs in the standing position were measured. The relation between the convex side of scoliosis and clinical parameters in terms of the side of symptoms, age, gender, duration of low back pain or leg pain, the angle of a positive straight leg raising test, and the time required for recovery of sciatic scoliosis were investigated. In addition, magnetic resonance imaging also was performed in five recent cases from 40 patients. The average Cobb angle decreased from 10.7 degrees to 2.7 degrees within an average of 7.5 months after surgery. The preoperative Cobb angle of patients with disc herniation medial to the nerve root was significantly higher than that just beneath or lateral to the nerve root. Thirty-two of 40 patients (80.0%) had a lumbar disc herniation at the convex side of scoliosis, irrespective of the transverse location of the herniation. The time required for scoliosis disappearance in disc herniation located lateral to the nerve root tended to be longer than that for other types of disc herniation. Magnetic resonance imaging through the paramedian planes showed enlargement of the intervertebral foramen at the convex side of scoliosis, compared with that at the concave side in five recent cases from the current study. These results suggest that sciatic scoliotic list is not a predictive factor of the anatomic location of disc herniation; rather, it is only suggestive of the side of disc herniation. The location of disc herniation may aide in the preoperative estimation of the recovery of the scoliosis.


Accurate scoring of the apnea-hypopnea index using a simple non-contact breathing sensor.
Sleep apnea is a serious condition that afflicts many individuals and is associated with serious health complications. Polysomnography, the gold standard for assessing and diagnosing sleep apnea, uses breathing sensors that are intrusive and can disrupt the patient's sleep during the overnight testing. We investigated the use of breathing signals derived from non-contact force sensors (i.e. load cells) placed under the supports of the bed as an alternative to traditional polysomnography breathing sensors (e.g. nasal pressure, oral-nasal thermistor, chest belt and abdominal belt). The apnea-hypopnea index estimated using the load cells was not different than that estimated using standard polysomnography leads (t44 = 0.37, P = 0.71). Overnight polysomnography sleep studies scored using load cell breathing signals had an intra-class correlation coefficient of 0.97 for the apnea-hypopnea index and an intra-class correlation coefficient of 0.85 for the respiratory disturbance index when compared with scoring using traditional polysomnography breathing sensors following American Academy of Sleep Medicine guidelines. These results demonstrate the feasibility of using unobtrusive load cells installed under the bed to measure the apnea-hypopnea index.


Structure of oxalacetate acetylhydrolase, a virulence factor of the chestnut blight fungus.
Oxalacetate acetylhydrolase (OAH), a member of the phosphoenolpyruvate mutase/isocitrate lyase superfamily, catalyzes the hydrolysis of oxalacetate to oxalic acid and acetate. This study shows that knock-out of the oah gene in Cryphonectria parasitica, the chestnut blight fungus, reduces the ability of the fungus to form cankers on chestnut trees, suggesting that OAH plays a key role in virulence. OAH was produced in Escherichia coli and purified, and its catalytic rates were determined. Oxalacetate is the main OAH substrate, but the enzyme also acts as a lyase of (2R,3S)-dimethyl malate with approximately 1000-fold lower efficacy. The crystal structure of OAH was determined alone, in complex with a mechanism-based inhibitor, 3,3-difluorooxalacetate (DFOA), and in complex with the reaction product, oxalate, to a resolution limit of 1.30, 1.55, and 1.65 A, respectively. OAH assembles into a dimer of dimers with each subunit exhibiting an (alpha/beta)(8) barrel fold and each pair swapping the 8th alpha-helix. An active site "gating loop" exhibits conformational disorder in the ligand-free structure. To obtain the structures of the OAH.ligand complexes, the ligand-free OAH crystals were soaked briefly with DFOA or oxalacetate. DFOA binding leads to ordering of the gating loop in a conformation that sequesters the ligand from the solvent. DFOA binds in a gem-diol form analogous to the oxalacetate intermediate/transition state. Oxalate binds in a planar conformation, but the gating loop is largely disordered. Comparison between the OAH structure and that of the closely related enzyme, 2,3-dimethylmalate lyase, suggests potential determinants of substrate preference.


Novel vascular effects of isoprenaline following pacing-induced heart failure in the dog.
Isoprenaline produced concentration-dependent contractions of the in vitro canine saphenous vein which were attenuated by phentolamine (10(-6) M) and pacing-induced heart failure. Both at control and peak heart failure, a biphasic response was seen in the dorsal pedal artery, consisting of an initial relaxation followed by a contraction; phentolamine and heart failure enhanced the relaxation component. In the presence of propranolol, the isoprenaline-induced contraction was sensitive to yohimbine, but resistant to prazosin. Therefore it is concluded that isoprenaline interacts not only with vascular beta-adrenoceptors, but also with alpha 2-adrenoceptors. Endothelial denudation resulted in a diminished response to isoprenaline in control saphenous vein and dorsal pedal artery but not in vessels from dogs with heart failure. The observation that the contractile response to isoprenaline diminishes in heart failure implies a specific down-regulation of peripheral vascular alpha 2-adrenoceptors.


DMRforPairs: identifying differentially methylated regions between unique samples using array based methylation profiles.
Array based methylation profiling is a cost-effective solution to study the association between genome methylation and human disease & development. Available tools to analyze the Illumina Infinium HumanMethylation450 BeadChip focus on comparing methylation levels per locus. Other tools combine multiple probes into a range, identifying differential methylated regions (DMRs). These tools all require groups of samples to compare. However, comparison of unique, individual samples is essential in situations where larger sample sizes are not possible. DMRforPairs was designed to compare regional methylation status between unique samples. It identifies probe dense genomic regions and quantifies/tests their (difference in) methylation level between the samples. As a proof of concept, DMRforPairs is applied to public data from four human cell lines: two lymphoblastoid cell lines from healthy individuals and the cancer cell lines A431 and MCF7 (including 2 technical replicates each). DMRforPairs identified an increasing number of DMRs related to the sample phenotype when biological similarity of the samples decreased. DMRs identified by DMRforPairs were related to the biological origin of the cell lines. To our knowledge, DMRforPairs is the first tool to identify and visualize relevant and significant differentially methylated regions between unique samples.


A Phase 4, Open-Label, Single-Arm Study Assessing the Efficacy and Safety of Ivabradine in African American Patients with Heart Failure and Reduced Ejection Fraction.
There are limited data on ivabradine therapy in black patients and none in African Americans. We performed an open-label, prospective study at two centers in the United States. African American patients with heart failure (HF) (N = 30), left ventricular ejection fraction ≤ 35%, and in sinus rhythm with resting heart rate (HR) ≥ 70 bpm received ivabradine 2.5-7.5 mg twice daily for 57 days. The primary endpoint was change in HR from baseline to day 57, compared with the -5 bpm change observed in the absence of ivabradine in the placebo group of the SHIFT study. The safety endpoint was treatment-emergent adverse events (TEAEs). Exploratory endpoints were change from baseline to day 57 in 6-minute walk test (6MWT) distance, HR difference during a 6MWT (i.e. HR at minute 6 - resting HR), and physical activity counts. At day 57, the estimated least squares mean change from baseline in HR was -9.5 bpm (95% CI -13.0, -6.0). The estimated mean treatment difference with ivabradine versus a presumed -5 bpm change from baseline HR, as seen in the placebo group of the SHIFT study, was -4.5 bpm (95% CI -8.0, -1.0; p = 0.013). The mean (SE) changes in 6MWT distance and HR difference during the 6MWT were 16.3 (10.8) meters and 2.3 (3.7) bpm, respectively. Ivabradine therapy did not result in greater physical activity. TEAEs were reported in 11 (36.7%) patients. These data support ivabradine use in African American patients with HF with reduced ejection fraction who meet typical treatment criteria. ClinicalTrials.gov identifier, NCT03456856.


Outcomes of bedside sutureless umbilical closure without endotracheal intubation for gastroschisis repair in surgical infants.
Newborns with gastroschisis have historically undergone surgical repair under general anesthesia. Our institution recently transitioned to the sutureless umbilical closure for gastroschisis. We sought to evaluate the feasibility of bedside gastroschisis repair without endotracheal intubation. A retrospective review was performed of neonates with gastroschisis who underwent sutureless umbilical closure from 2011 to 2015. Clinical characteristics and outcomes between groups were compared. In total, 53 infants underwent sutureless umbilical closure. Closure without endotracheal intubation was attempted in 23 (43%) babies and was successful in 15 (65%) infants. Two of the 8 patients who required intubation needed a temporary silo. Neonates successfully repaired without intubation were more premature (p < 0.01), smaller at birth (p = 0.01), and repaired nearly an hour sooner (p < 0.01). There were no differences in time to full enteral nutrition, length of stay, bowel ischemia, or sepsis. Bedside sutureless umbilical closure without intubation is feasible and effective in newborns with gastroschisis. The procedure decreases time to gastroschisis closure. Smaller and more premature neonates were more likely to be successfully closed without intubation.


Preferential adsorption and activity of monocomponent cellulases on lignocellulose thin films with varying lignin content.
Understanding the enzymatic hydrolysis of cellulose and the influence of lignin in the process are critical for viable production of fuels and chemicals from lignocellulosic biomass. The interactions of monocomponent cellulases with cellulose and lignin substrates were investigated by using thin films supported on quartz crystal microgravimetry (QCM) resonators. Trichoderma reesei exoglucanase (CBH-I) and endoglucanase (EG-I) bound strongly to both cellulose and lignin but EG-I exhibited a distinctive higher affinity with lignin, causing a more extensive inhibition of the cellulolytic reactions. CBH-I was found to penetrate into the bulk of the cellulose substrate increasing the extent of hydrolysis and film deconstruction. In the absence of a cellulose binding domain (CBD) and a linker, the CBH-I core adsorbed slowly and was not able to penetrate into the film. Conversely to CBH-I, EG-I exhibited activity only on the surface of the lignocellulose substrate even when containing a CBD and a linker. Interestingly, EG-I displayed a clearly different interaction profile as a function of contact time registered by QCM.


Folate grafted thiolated chitosan enveloped nanoliposomes with enhanced oral bioavailability and anticancer activity of docetaxel.
Folate grafted and thiolated chitosan was synthesized and wrapped on the surface of mixed phosphatidylcholine based nanoliposomes (NLs) to improve the oral absorption and targeted pharmacological activity of anti-cancer drugs against breast cancer. In this study, a chitosan derived thiomer, having intrinsic properties of P-glycoprotein (P-gp) efflux pump inhibition, mucoadhesion and controlled drug release at a target site, was exploited to improve the performance of docetaxel (DTX) loaded NLs for better oral pharmacokinetics, targeted anti-cancer activity, liposomal stability and the physical characteristics of NLs. Thiomer enveloped nanoliposomes (ENLs) and bare nanoliposomes (NLs) were synthesized with the ingredient ratio pre-determined via Response Surface Methodology (RSM) plots by Design Expert® software. ENLs and NLs were thoroughly characterized for their surface chemistry, particle size, zeta potential, PDI, encapsulation efficiency, stability and release profile. ENLs were spherical in shape with a particle size of 328.5 ± 30 nm, a positive zeta potential of 18.81 ± 2.45 and a high encapsulation efficiency of 83% for DTX. Controlled release of DTX from formulations was observed for over 72 h for each formulation. The presence of thiol groups at the surface of the ENLs resulted in higher swelling and in situ gelling properties compared to the corresponding NLs. Furthermore, ENL/mucin mixtures showed a time dependent increase in viscosity for up to 12 h, leading to a 19.07-fold increased viscosity. Ex vivo permeation and P-glycoprotein inhibiting properties, studied in rat's small intestine, showed a 9.6-fold higher permeation and 13-fold enhancement of DTX in the presence of ENLs. In vitro cytotoxicity studies indicated that the ENLs can efficiently kill MD-MB-231 breast cancer cells with 200 fold lower IC50 values than DTX alone as a positive control. The pharmacokinetic study revealed that the ENLs significantly improved the oral bioavailability of DTX i.e. up to 13.6 fold as compared to an aqueous dispersion of DTX. Therefore, these enveloped hybrid nanoliposomes (ENLs) have the potential to be developed as useful nanocarriers for efficient oral delivery and breast cancer management using DTX.


[Stem cells and prenatal therapy].
The special biological properties of stem cells bring us new hope for the research and application of prenatal therapy. The basic research of prenatal therapy utilizing stem cells developed quickly,providing reference and direction for clinical therapy. This review is mainly related to the progress in this field in recent years.


Microplastics in freshwater systems: a review of the emerging threats, identification of knowledge gaps and prioritisation of research needs.
Plastic contamination is an increasing environmental problem in marine systems where it has spread globally to even the most remote habitats. Plastic pieces in smaller size scales, microplastics (particles <5 mm), have reached high densities (e.g., 100,000 items per m(3)) in waters and sediments, and are interacting with organisms and the environment in a variety of ways. Early investigations of freshwater systems suggest microplastic presence and interactions are equally as far reaching as are being observed in marine systems. Microplastics are being detected in freshwaters of Europe, North America, and Asia, and the first organismal studies are finding that freshwater fauna across a range of feeding guilds ingest microplastics. Drawing from the marine literature and these initial freshwater studies, we review the issue of microplastics in freshwater systems to summarise current understanding, identify knowledge gaps and suggest future research priorities. Evidence suggests that freshwater systems may share similarities to marine systems in the types of forces that transport microplastics (e.g. surface currents); the prevalence of microplastics (e.g. numerically abundant and ubiquitous); the approaches used for detection, identification and quantification (e.g. density separation, filtration, sieving and infrared spectroscopy); and the potential impacts (e.g. physical damage to organisms that ingest them, chemical transfer of toxicants). Differences between freshwater and marine systems include the closer proximity to point sources in freshwaters, the typically smaller sizes of freshwater systems, and spatial and temporal differences in the mixing/transport of particles by physical forces. These differences between marine and freshwater systems may lead to differences in the type of microplastics present. For example, rivers may show a predictable pattern in microplastic characteristics (size, shape, relative abundance) based on waste sources (e.g. household vs. industrial) adjacent to the river, and distance downstream from a point source. Given that the study of microplastics in freshwaters has only arisen in the last few years, we are still limited in our understanding of 1) their presence and distribution in the environment; 2) their transport pathways and factors that affect distributions; 3) methods for their accurate detection and quantification; 4) the extent and relevance of their impacts on aquatic life. We also do not know how microplastics might transfer from freshwater to terrestrial ecosystems, and we do not know if and how they may affect human health. This is concerning because human populations have a high dependency on freshwaters for drinking water and for food resources. Increasing the level of understanding in these areas is essential if we are to develop appropriate policy and management tools to address this emerging issue.


The relationship between magnitude of proteinuria reduction and risk of end-stage renal disease: results of the African American study of kidney disease and hypertension.
The magnitude of proteinuria is associated with a graded increase in the risk of progression to end-stage renal disease and cardiovascular events. The objective of this study was to relate baseline and early changes in proteinuria and glomerular filtration rate (GFR) to long-term progression of hypertensive nondiabetic kidney disease. Post hoc analysis of a randomized 3 x 2 factorial trial. A total of 1094 African Americans with hypertensive renal disease (GFR, 20-65 mL/min per 1.73 m(2)) were followed up for a median of 3.8 years. Participants were randomized to a mean arterial pressure goal of 102 to 107 mm Hg (usual) or 92 mm Hg or less (lower) and to initial treatment with a beta-blocker (metoprolol), an angiotensin-converting enzyme inhibitor (ramipril), or a dihydropyridine calcium channel blocker (amlodipine) Baseline proteinuria and GFR predicted the rgate of GFR decline. For each 10-mL/min per 1.73 m(2) lower baseline GFR, an associated mean +/- SE 0.38 +/- 0.08-mL/min per 1.73 m(2) per year greater mean GFR decline occurred, and for each 2-fold higher proteinuria level, a mean +/- SE 0.54 +/- 0.05-mL/min per 1.73 m(2) per year faster GFR decline was observed (P < .001 for both). In multivariate analysis, the effect of baseline proteinuria GFR decline persisted. Initial change in proteinuria from baseline to 6 months predicted subsequent progression, with this relationship extending to participants with baseline urinary protein levels less than 300 mg/d. The change in the level of proteinuria is a predictor of subsequent progression of hypertensive kidney disease at a given GFR. A prospective trial is needed to confirm this observation.


Medical education at the University of Wisconsin-Madison: comparison of medical student and resident education as to the number of physicians practicing in the state of Wisconsin.
The primary mission of a state-supported medical education is to produce physicians who will practice in that state. Medical school and residency graduates at the University of Wisconsin-Madison were compared as to how often they practice in the state after completing training. Six hundred ninety-three medical student graduates from 1987 to 1991 were compared with 657 residency graduates from 1992 to 1996 at the University of Wisconsin-Madison. Chi-square was used to compare the groups as to the number of physicians produced who: 1) practice in Wisconsin and 2) practice primary care in Wisconsin. The residency training programs produced significantly more physicians (280) who practice in Wisconsin than did the medical school (246), X2 (df = 1) = 7.20, p < .01, and also significantly more primary care physicians, X2 (df = 1) = 6.16, p < .02. When this evidence-based information is used as a measure of medical education outcomes, residency training may be more effective at producing practicing physicians and should not be discounted when planning the educational and public health needs for the state of Wisconsin.


Public and physicians' support for euthanasia in people suffering from psychiatric disorders: a cross-sectional survey study.
Although euthanasia and assisted suicide (EAS) in people with psychiatric disorders is relatively rare, the increasing incidence of EAS requests has given rise to public and political debate. This study aimed to explore support of the public and physicians for euthanasia and assisted suicide in people with psychiatric disorders and examine factors associated with acceptance and conceivability of performing EAS in these patients. A survey was distributed amongst a random sample of Dutch 2641 citizens (response 75%) and 3000 physicians (response 52%). Acceptance and conceivability of performing EAS, demographics, health status and professional characteristics were measured. Multivariable logistic regression analyses were performed. Of the general public 53% were of the opinion that people with psychiatric disorders should be eligible for EAS, 15% was opposed to this, and 32% remained neutral. Higher educational level, Dutch ethnicity, and higher urbanization level were associated with higher acceptability of EAS whilst a religious life stance and good health were associated with lower acceptability. The percentage of physicians who considered performing EAS in people with psychiatric disorders conceivable ranged between 20% amongst medical specialists and 47% amongst general practitioners. Having received EAS requests from psychiatric patients before was associated with considering performing EAS conceivable. Being female, religious, medical specialist, or psychiatrist were associated with lower conceivability. The majority (> 65%) of the psychiatrists were of the opinion that it is possible to establish whether a psychiatric patient's suffering is unbearable and without prospect and whether the request is well-considered. The general public shows more support than opposition as to whether patients suffering from a psychiatric disorder should be eligible for EAS, even though one third of the respondents remained neutral. Physicians' support depends on their specialization; 39% of psychiatrists considered performing EAS in psychiatric patients conceivable. The relatively low conceivability is possibly explained by psychiatric patients often not meeting the eligibility criteria.


Residential (inpatient) treatment compared with outpatient treatment for nicotine dependence.
To compare smoking abstinence outcomes between smokers treated in a residential (inpatient) program and those treated in an outpatient program to determine if residential treatment was superior to outpatient treatment in smokers with moderate to severe nicotine dependence. Patients treated in the residential nicotine dependence program at the Mayo Clinic, Rochester, Minn., between May 1, 1992, and January 31, 1996, were selected for this study. Each patient in the residential treatment group (n=146) was matched to 2 patients who received an outpatient nicotine dependence consultation by a trained counselor (n=292). Each patient was matched on age, sex, year seen, number of cigarettes smoked per day, longest previous abstinence, education, and marital status. Abstinence at 6 and 12 months was determined by self-report. For the purposes of analysis, each patient with missing outcome data was considered to be smoking. The 6-month abstinence rates for the residential group compared with the outpatient group were 45% and 26%, respectively (P<.001), and the 12-month abstinence rates were 45% and 23%, respectively (P<.001). After adjusting for matching variables that were not exactly matched (age, baseline number of cigarettes smoked per day, and longest previous abstinence) and the baseline variables, including education, age when started smoking, and degree of nicotine dependence, there was a significant effect of residential treatment on 6- and 12-month abstinence rates (P<.001). Odds ratio of 6-month abstinence in the residential group was 2.74 (95% confidence interval, 1.60-4.71; P<.001) and at 12 months was 3.03 (95% confidence interval, 1.74-5.27; P<.001). Residential treatment for tobacco dependence is superior to outpatient treatment in some smokers who are moderately to severely nicotine dependent.


Feature Learning Networks for Floor Sensor-based Gait Recognition.
Deep learning (DL) has become a powerful tool in many image classification applications but often requires large training sets to achieve high accuracy. For applications where the available data are limited, this can become a severely limiting factor in model performance. To address this limitation, feature learning network approaches that integrate traditional feature extraction methods with DL frameworks have been proposed. In this study, the performances of traditional methods: discrete wavelet transform (DWT), discrete cosine transform (DCT), independent component analysis (ICA), and principal component analysis (PCA); and their corresponding feature networks based on a convolutional neural network (CNN) framework: ScatNet (wavelet scattering network), DCTNet, ICANet, and PCANet, were investigated for use in pressure-based footstep recognition when the limited sample size is available for person authentication. The results show that the feature learning networks (90.6% accuracy) achieved significantly better performance on average than the conventional feature extraction methods (79.7% accuracy) (p < 0.05). Among the different feature networks, PCANet provided the best verification performance, with an accuracy of 92.2%. Feature learning networks are simple and effective approaches that can be a promising solution for applications like floor-based gait recognition in a security access scenario (such as workspace environment and border control) when small amounts of data are available for training models to differentiate between a larger group of users.


Innate interferons inhibit allergen and microbial specific T(H)2 responses.
Several studies provided evidence of innate interferons (IFNs) regulating T(H)2 cytokine production using purified CD4(+) memory cells and T(H)2 polarisation via interleukin-4 (IL-4). Vitally, none of these previous studies examined IFN attenuation of T(H)2 responses to allergen or antigen. This study therefore sought to investigate the abrogation of specific allergen- and antigen-stimulated T(H)2 response in peripheral blood mononuclear cells (PBMC) derived from 12 sensitised individuals by IFN-β and IFN-λ. PBMC were cultured in the presence of house dust mite (HDM) allergen, rhinovirus (RV), influenza vaccine and tetanus toxoid (TT)±either IFN-β or IFN-λ for 3 and 5 days. IFN-γ, IL-5 and IL-13 protein levels were measured by ELISA. Quantitative PCR (qPCR) was used to investigate induction of genes involved in control of T(H)2 cytokines. No alteration in T(H)1 IFN-γ allergen/antigen response was observed with addition of IFN-β or IFN-λ. Consistent abrogation of T(H)2 response to HDM and influenza was observed with IFN-β at both time points; attenuation was observed by day 5 with RV and TT. IFN-λ had no consistent effect on T(H)2 production except in the presence of RV (multiplicity of infection=5); a decrease in IL-5 alone was observed in the presence of trivalent inactivated influenza vaccine. GATA binding protein 3 (GATA3) and suppressors of cytokine signalling3 mRNA were differentially regulated in HDM and influenza-stimulated cultures±IFN-β. We concluded that IFN-β produced a strong and consistent abrogation of T(H)2 cytokine production in the presence of a range of allergen and antigen stimulants.


Neural stem cells, the subventricular zone and radiotherapy: implications for treating glioblastoma.
Over the past decade, advances in neuroscience have suggested that neural stem cells resident in specific regions of the adult brain may be involved in development of both primary and recurrent glioblastoma. Neurogenesis and malignant transformation occurs in the subventricular zone adjacent to the lateral ventricles. This region holds promise as a potential target for therapeutic intervention with radiotherapy. However, irradiation of a larger brain volume is not without risk, and significant side effects have been observed. The current literature remains contradictory regarding the efficacy of deliberate intervention with radiation to the subventricular zone. This critical review discusses the connection between neural stem cells and development of glioblastoma, explores the behavior of tumors associated with the subventricular zone, summarizes the discordant literature with respect to the effects of irradiation, and reviews other targeted therapies to this intriguing region.


High foetal mortality and birth intervals.
Abstract Studies of birth intervals have generally separated the process into: (1) the period of infertility following a pregnancy termination; (2) the length ofthe interval from the end of the infertile period to the next conception (which reflects the probability of conception), and (3) the average duration of pregnancy for both live births and foetal deaths.(1) Empirically derived intervals between successive live births average from 19 to 30 lunar months(2) (Table 1), depending upon the population studied and the birth order.(3) Estimates of conception rates using birth intervals data require some estimate of the extent to which foetal wastage increases the birth interval.


The role of the mussel Mytilus spp. in the transmission of ostreid herpesvirus-1 microVar.
The Pacific oyster Crassostrea gigas contributes significantly to global aquaculture; however, C. gigas culture has been affected by ostreid herpesvirus-1 (OsHV-1) and variants. The dynamics of how the virus maintains itself at culture sites is unclear and the role of carriers, reservoirs or hosts is unknown. Both wild and cultured mussels Mytilus spp. (Mytilus edulis, Mytilus galloprovincialis and hybrids) are commonly found at C. gigas culture sites. The objective of this study was to investigate if Mytilus spp. can harbour the virus and if viral transmission can occur between mussels and oysters. Mytilus spp. living at oyster trestles, 400-500 m higher up the shore from the trestles and up to 26 km at non-culture sites were screened for OsHV-1 and variants by all the World Organization for Animal Health (OIE) recommended diagnostic methods including polymerase chain reaction (PCR), quantitative PCR (qPCR), histology, in situ hybridization and confirmation using direct sequencing. The particular primers that target OsHV-1 and variants, including OsHV-1 microVar (μVar), were used in the PCR and qPCR. OsHV-1 μVar was detected in wild Mytilus spp. at C. gigas culture sites and more significantly the virus was detected in mussels at non-culture sites. Cohabitation of exposed wild mussels and naïve C. gigas resulted in viral transmission after 14 days, under an elevated temperature regime. These results indicate that mussels can harbour OsHV-1 μVar; however, the impact of OsHV-1 μVar on Mytilus spp. requires further investigation.


Absence of covalently linked core protein from newly synthesized hyaluronate.
1. Primary cultures of chondrocytes from the Swarm rat chondrosarcoma were labelled with either [3H]glucosamine or [14C]glucosamine, and hyaluronate synthesized by the cells was isolated from the cell layer. Parallel cultures were labelled with either [3H]serine or [3H]lysine, and identical fractions were isolated from the cell layer. Some cultures were dual-labelled. 2. In cultures labelled with [3H]serine for between 30 min and 24 h and extracted with 4.0 M-guanidine, a procedure that solubilizes predominantly extracellular macromolecules, small amounts of [3H]serine-labelled molecules were found associated with the hyaluronate fraction purified from the extract by dissociative CsCl-density-gradient centrifugation and dissociative Sepharose CL-2B chromatography. About 75% of the [3H]serine-labelled molecules in the fraction were specifically associated with hyaluronate, since they could be removed by prior treatment with proteinase-free Streptomyces hyaluronidase. The association of the [3H]serine-labelled molecules with hyaluronate was non-covalent, since they could be separated from it by further centrifugation in CsCl density gradients containing 4 M-guanidinium chloride and a zwitterionic detergent. 3. In other experiments the cultures were extracted with a sequential zwitterionic-detergent/guanidinium chloride procedure that completely solubilized the cell layer and enabled fractions containing newly synthesized cell-associated hyaluronate to be isolated. Zwitterionic detergent was present throughout. No [3H]lysine was incorporated into these fractions, irrespective of whether the cultures were pulsed concurrently with [3H]lysine and [14C]glucosamine or sequentially with [3H]lysine to prelabel the protein pool (24 h) followed by [14C]-glucosamine to label hyaluronate (1 h). 4. The results show that newly synthesized hyaluronate is not associated with covalently bound protein, and suggest that chain synthesis is initiated by a mechanism other than on to a core protein. Small amounts of [3H]serine-labelled molecules are, however, non-covalently associated with extracellular hyaluronate. Their identity is at present unknown, but they are probably of low molecular weight.


Efficiency of different air filter types for pig facilities at laboratory scale.
Air filtration has been shown to be efficient in reducing pathogen burden in circulating air. We determined at laboratory scale the retention efficiency of different air filter types either composed of a prefilter (EU class G4) and a secondary fiberglass filter (EU class F9) or consisting of a filter mat (EU class M6 and F8-9). Four filter prototypes were tested for their capability to remove aerosol containing equine arteritis virus (EAV), porcine reproductive and respiratory syndrome virus (PRRSV), bovine enterovirus 1 (BEV), Actinobacillus pleuropneumoniae (APP), and Staphylococcus (S.) aureus from air. Depending on the filter prototype and utilisation, the airflow was set at 1,800 m3/h (combination of upstream prefilter and fiberglass filter) or 80 m3/h (filter mat). The pathogens were aerosolized and their concentration was determined in front of and behind the filter by culture or quantitative real-time RT-PCR. Furthermore, survival of the pathogens over time in the filter material was determined. Bacteria were most efficiently filtered with a reduction rate of up to 99.9% depending on the filter used. An approximately 98% reduction was achieved for the viruses tested. Viability or infectivity of APP or PRRSV in the filter material decreased below the detection limit after 4 h and 24 h, respectively, whereas S. aureus was still culturable after 4 weeks. Our results demonstrate that pathogens can efficiently be reduced by air filtration. Consequently, air filtration combined with other strict biosecurity measures markedly reduces the risk of introducing airborne transmitted pathogens to animal facilities. In addition, air filtration might be useful in reducing bioaerosols within a pig barn, hence improving respiratory health of pigs.


[The influencing factors on suicide attempt among adolescents in South Korea].
Suicide has been considered a large public health concern in recent years because suicide mortality has been increasing rapidly. This study was done to investigate the prevalence and risk factors of a suicide attempt among adolescents in South Korea. The data of the 2006 Youth Health Risk Behavior web-based Survey collected by the Korean Center for Disease Control was analyzed using logistic regression for this study. The prevalence of a suicide attempt was 5.2% in South Korea. The risk factors of a suicide attempt were suicidal ideation (odds 31.83), depression (odds 7.98), drug use (odds 4.67), currently smoking (odds 3.19), feeling unhappiness (odds 2.77), stress (odds 2.60), currently drinking alcohol (odds 2.39), sexual activity (odds 2.33), living with neither parent (odds 2.24), initial alcohol drinking by age 9 (odds 1.80), health status (odds 2.15), skipped breakfast (odds 1.75), disease (odds 1.65), and school records (odds 1.22). Suicide screening and prevention programs should be developed, considering these risk factors. Suicide prevention programs should be applied to the at risk population prior to a suicide attempt.


A systematic assessment on COVID-19 preparedness and transition strategy in Bangladesh.
The COVID-19 pandemic of 2020 has overpowered the most advanced health systems worldwide with thousands of daily deaths. The current study conducted a situation analysis on the pandemic preparedness of Bangladesh and provided recommendations on the transition to the new reality and gradual restoration of normalcy. A complex adaptive system (CAS) framework was theorized based on four structural dimensions obtained from the crisis and complexity theory to help evaluate the health system of Bangladesh. Data sourced from published reports from the government, non-governmental organizations, and mainstream media up to June 15, 2020 were used to conduct a qualitative analysis and visualize the spatial distribution of countrywide COVID-19 cases. The findings suggested that Bangladesh severely lacked the preparedness to tackle the spread of COVID-19 with both short- and long-term implications for health, the economy, and good governance. Absence of planning and coordination, disproportionate resource allocations, challenged infrastructure, adherence to bureaucratic delay, lack of synchronized risk communication, failing leadership of concerned authorities, and incoherent decision-making have led to a precarious situation that will have dire ramifications causing many uncertainties in the coming days. Implementation of response protocols addressing the needs of the community and the stakeholders from the central level is urgently needed. The development of mechanisms for dynamic decision-making based on regular feedback and long-term planning for a smooth transition between the new reality and normalcy should also be urgently addressed in Bangladesh.


Gestational exposure to 1-NP induces ferroptosis in placental trophoblasts via CYP1B1/ERK signaling pathway leading to fetal growth restriction.
Fetal growth restriction (FGR) is a prevalent complication in obstetrics, yet its exact aetiology remains unknown. Numerous studies suggest that the degradation of the living environment is a significant risk factor for FGR. 1-Nitropyrene (1-NP) is a widespread environmental pollutant as a representative substance of nitro-polycyclic aromatic hydrocarbons. In this study, we revealed that 1-NP induced FGR in fetal mice by constructing 1-NP exposed pregnant mice models. Intriguingly, we found that placental trophoblasts of 1-NP exposed mice exhibited significant ferroptosis, which was similarly detected in placental trophoblasts from human FGR patients. In this regard, we established a 1-NP exposed cell model in vitro using two human trophoblast cell lines, HTR8/SVneo and JEG-3. We found that 1-NP not only impaired the proliferation, migration, invasion and angiogenesis of trophoblasts, but also induced severe cellular ferroptosis. Meanwhile, the ferroptosis inhibitor ferrostatin-1 (Fer-1) effectively rescued 1-NP-induced trophoblast biological function impairment. Mechanistically, we revealed that 1-NP regulated ferroptosis by activating the ERK signaling pathway. Moreover, we innovatively revealed that CYP1B1 was essential for the activation of ERK signaling pathway induced by 1-NP. Overall, our study innovatively identified ferroptosis as a significant contributor to 1-NP induced trophoblastic functional impairment leading to FGR and clarified the specific mechanism by which 1-NP induced ferroptosis via the CYP1B1/ERK signaling pathway. Our study provided novel insights into the aetiology of FGR and revealed new mechanisms of reproductive toxicity of environmental pollutants.


Effect of bipolar disorder on lymphocyte inositol monophosphatase mRNA levels.
The activity of inositol monophosphatase (IMPase), the lithium (Li)-inhibitable enzyme in the phosphatidylinositol (PI) signal transduction system, has recently been found significantly lower in lymphoblastoid cell lines from bipolar (BP) patients, particularly in Li-responders. To probe for possible quick detection of the disease and prediction of the therapeutic response we repeated our study in fresh lymphocytes. Since IMPase in fresh lymphocytes is inhibited in vivo by ongoing Li treatment and its pre-Li activity cannot be evaluated, IMPase mRNA levels were measured. Relative (to beta-actin) mRNA levels were quantified by reverse transcriptase (RT)-PCR in 5 drug-free and 31 drug-treated BP patients compared with 36 control subjects in fresh lymphocytes. In agreement with our findings with IMPase activity, the small group of drug-free BP patients exhibited approximately 2/3 reduction in IMPase relative mRNA levels compared to control subjects. Approximately 2-fold elevation of these levels toward control values was found for patients treated with Li and other mood stabilizers. The study further suggests the possible importance of IMPase in the aetiology of BP disorder and in the mediation of the therapeutic efficacy of Li. It may be that chronic inhibition of IMPase activity by Li results in up-regulation of its gene at the transcriptional level.


Continuous cardiac output and mixed venous oxygen saturation monitoring.
Continuous assessment of cardiac output and SVO2 in the critically ill may be helpful in both the monitoring variations in the patient's cardiovascular state and in determining the efficacy of therapy. Commercially available continuous cardiac output (CCO) monitoring systems are based on the pulsed warm thermodilution technique. In vitro validation studies have demonstrated that this method provides higher accuracy and greater resistance to thermal noise than standard bolus thermodilution techniques. Numerous clinical studies comparing bolus with continuous thermodilution techniques have shown this technique similarly accurate to track each other and to have negligible bias between them. The comparison between continuous thermal and other cardiac output methods also demonstrates good precision of the continuous thermal technique. Accuracy of continuous oximetry monitoring using reflectance oximetry via fiberoptics has been assessed both in vitro and in vivo. Most of the studies testing agreement between continuous SVO2 measurements and pulmonary arterial blood samples measured by standard oximetry have shown good correlation. Continuous SVO2 monitoring is often used in the management of critically ill patients. The most recently designed pulmonary artery catheters are now able to simultaneously measure either SVO2 and CCO or SVO2 and right ventricular ejection fraction. This ability to view simultaneous trends of SVO2 and right ventricular performance parameters will probably allow the clinician to graphically see the impact of volume loading or inotropic therapy over time, as well as the influence of multiple factors, including right ventricular dysfunction, on SVO2. However, the cost-effectiveness of new pulmonary artery catheters application remains still questionable because no established utility or therapeutic guidelines are available.


Screening of congenital cytomegalovirus infection by real-time PCR in urine pools.
Human cytomegalovirus (HCMV) is the most frequent cause of congenital infection. Despite the fact that 5-17% of asymptomatic infected babies will develop late sequelae and, therefore, should be closely followed, most of these children will remain undetected, as screening of all newborns by viral culture is too expensive and no valid alternative has been widely accepted. The aim of this work was to demonstrate that pool testing can be used to screen HCMV congenital infection in newborns. For this purpose, a real-time PCR technique was tested in urine pools. This pool method was applied to all urine specimens from children received in the virology laboratory of the Centro Hospitalar Cova da Beira for diagnosis of HCMV infection for a period of 14 months. Ten out of the 160 urine samples were tested positive by shell-vial culture and were also detected by this pool method. Additionally, 100 urine specimens, collected in 2004 and culture negative for HCMV were included to test the specificity of this methodology, all of which were negative. In conclusion, these results suggest that urine pools can be used to detect HCMV-positive urines in children, with similar sensitivity and specificity when compared with the standard method. Because of the very significant reduction both in terms of labour and cost of testing materials, this methodology may represent a valid option for screening the HCMV congenital infection in newborns.


Ca(2+) -induced binding of anticoagulation factor II from the venom of Agkistrodon acutus with factor IX.
Anticoagulation factor II (ACF II), a coagulation factor X- binding protein from the venom of Agkistrodon acutus has both anticoagulant and hypotensive activities. Previous studies show that ACF II binds specifically with activated factor X (FXa) in a Ca(2+) -dependent manner and inhibits intrinsic coagulation pathway. In this study, the inhibition of extrinsic coagulation pathway by ACF II was measured in vivo by prothrombin time assay and the binding of ACF II to factor IX (FIX) was investigated by native polyacrylamide gel electrophoresis and surface plasmon resonance (SPR). The results indicate that ACF II also inhibits extrinsic coagulation pathway, but does not inhibit thrombin activity. ACF II also binds with FIX with high binding affinity in a Ca(2+) -dependent manner and their maximal binding occurs at about 0.1 mM Ca(2+) . ACF II has similar binding affinity to FIX and FX as determined by SPR. Ca(2+) has a slight effect on the secondary structure of FIX as determined by circular dichroism spectroscopy. Ca(2+) ions are required to maintain in vivo function of FIX Gla domain for its recognition of ACF II. However, Ca(2+) at high concentrations (>0.1 mM) inhibits the binding of ACF II to FIX. Ca(2+) functions as a switch for the binding between ACF II and FIX. ACF II extends activated partial thromboplastin time more strongly than prothrombin time, suggesting that the binding of ACF II with FIX may play a dominant role in the anticoagulation of ACF II in vivo.


Crystal Structure Dependent Dissolution of Non-Cubic Au Crystallites in Aqua Regia.
Properties of metal crystallites are governed by their morphologies and inherent crystal structures. In this work, bipyramidal Au microcrystallites hosting non-cubic lattices, body-centered orthorhombic and tetragonal (together termed as bc(o,t)), are investigated for their stability in aqua regia. Specifically, microcrystallites comprising 92 % of bc(o,t) have been subjected to aqua regia of different concentrations and the changes in morphology and lattice phases have been monitored using scanning electron microscopy and X-ray diffraction techniques. The dissolution process was found to be crystal structure dependent and begin at the bipyramidal tips enriched with fcc lattice while retaining the bc(o,t) rich body. Interestingly, with increasing the reaction times, the remaining core was found to be highly reluctant to dissolution and instead, transformed to tetragonal lattices which with increasing treatment, exhibited lattice parameters closer to that of fcc. The study reveals the presence of a bc(o,t)-fcc core-shell structure with the tips enriched with fcc.


Tough gel adhesive is an effective method for meniscal repair in a bovine cadaveric study.
To test tough gel adhesives to repair meniscus tears under relevant loading conditions and determine if they have adequate biomechanical properties to repair meniscus tears in a bovine cadaveric study. Cyclic compression tests on 24 dissected bovine knees were performed. The tough gel adhesive was used either as an adhesive patch or as a coating bonded onto commercially available surgical sutures. Forty-eight menisci were tested in this study; 24 complete radial tears and 24 bucket-handle tears. After preconditioning, the specimens underwent 100 cycles of compression, (800 N/0.5 Hz) on an Instron© machine and the size of the gaps measured. One third of the menisci were repaired with pristine sutures, one third with adhesive patches, and one third with sutures coated in adhesive gel. The size of the gaps was compared after 100 and 500 cycles of compression. The mean gap measured at the tear site without treatment was 6.46 mm (± 1.41 mm) for radial tears and 1.92 mm (± 0.65 mm) for bucket-handle tears. After treatment and 500 cycles of compression, the mean gap was 1.63 mm (± 1.41 mm) for pristine sutures, 1.50 mm (± 1.16 mm) for adhesive sutures and 2.06 mm (± 1.53 mm) for adhesive gel patches. There was no significant difference between treatments regardless of the type of tear. Also, the gaps for radial tears increased significantly with the number of compression cycles applied (p > 0.001). From a biomechanical standpoint, the tough adhesive gel patch is as effective as suturing. In addition, it would allow the repair of non-suturable tears and thus broaden the indications for meniscus repair. Controlled laboratory study.


Somatic gene mutation and human disease other than cancer: an update.
Somatic mosaicism is well known in disorders where the manifestations are readily seen, e.g. the skin in neurofibromatosis I. In single gene disorders of higher frequency, especially X-linked ones, the frequency of combined germ-line and somatic mosaicism is increasingly being appreciated, e.g. Duchenne Muscular Dystrophy. Cell separation techniques; such as the fluorescence-activated cell sorter (FACS) also detect much somatic mosaicism among blood cells in disorders such as paroxysmal nocturnal hemoglobinuria. Depending on the disorder and the class of mutation, in genes for which there are sufficient numbers of patients studied, 6-20% of cases are due to somatic mutation. This update of my previous review is stimulated by the rapid application of new technologies for the study of DNA variation in disease. The results of these studies implicate somatic mutation in a greater variety of genetic diseases and a wider spectrum of tissues than have previously been shown, including heart and kidney. The classes of mutation have also expanded beyond base pair changes, insertions/deletion (indels), and short tandem repeat mutations to include copy number variants and transposon-mediated mutations. I also briefly discuss previously well-known mosaicism for chromosomal mutations. Genomic sequencing, performed on DNA from blood, shows many mutations which are conclusively somatic in origin. It is still too early to see if there is a different pattern of somatic mutation compared to germ-line mutation. Though the parameters to allow careful quantification are not yet available, it seems that the frequency of gene mutation in embryonic cells is not markedly different than that in the germ line.


Emotional and cognitive consequences of head injury in relation to the glasgow outcome scale.
There is current debate over the issue of the best way of assessing outcome after head injury. One criticism of scales of disability and handicap such as the Glasgow outcome scale (GOS) is that they fail to capture the subjective perspective of the person with head injury. The aims of the study were to investigate aspects of the validity of structured interviews for the GOS, and address the issue of the relation between the GOS and subjective reports of health outcome. A total of 135 patients with head injury were assessed using the GOS and an extended GOS (GOSE) and other measures of outcome and clinical status at 6 months after injury. There were robust correlations between the GOS and measures of initial injury severity (particularly post-traumatic amnesia) and outcome assessed by disability scales (particularly the disbility rating scale (DRS)); however, associations with cognitive tests were generally modest. There were also strong correlations with self report measures of health outcome: both the GOS and GOSE were related to depression measured by the Beck depression inventory, mental wellbeing assessed by the general health questionnaire, and to all subscales of the short form-36. The GOS scales were also strongly associated with frequency of reported symptoms and problems on the neurobehavioural functioning inventory. The GOS and GOSE show consistent relations with other outcome measures including subjective reports of health outcome; they thus remain useful overall summary assessments of outcome of head injury.


Seasonal Variation in Sea Turtle Density and Abundance in the Southeast Florida Current and Surrounding Waters.
Assessment and management of sea turtle populations is often limited by a lack of available data pertaining to at-sea distributions at appropriate spatial and temporal resolutions. Assessing the spatial and temporal distributions of marine turtles in an open system poses both observational and analytical challenges due to the turtles' highly migratory nature. Surface counts of marine turtles in waters along the southern part of Florida's east coast were made in and adjacent to the southeast portion of the Florida Current using standard aerial surveys during 2011 and 2012 to assess their seasonal presence. This area is of particular concern for sea turtles as interest increases in offshore energy developments, specifically harnessing the power of the Florida Current. While it is understood that marine turtles use these waters, here we evaluate seasonal variation in sea turtle abundance and density over two years. Density of sea turtles observed within the study area ranged from 0.003 turtles km-2 in the winter of 2011 to 0.064 turtles km-2 in the spring of 2012. This assessment of marine turtles in the waters off southeast Florida quantifies their in-water abundance across seasons in this area to establish baselines and inform future management strategies of these protected species.


Papillary muscle dyssynchrony as a cause of functional mitral regurgitation in non-ischemic dilated cardiomyopathy patients with narrow QRS complexes.
Mitral regurgitation (MR) increases mortality in dilated cardiomyopathy (DCM). We investigated the prevalence of functional MR in non-ischemic DCM patients with narrow QRS intervals and its association with papillary muscle dyssynchrony. Ninety-three patients were enrolled consecutively in this cross-sectional study. Patients were evaluated for the presence of intraventricular (DYS Sep-Lat Sys) and papillary muscle (DYS Inter PAP Sys) systolic dyssynchrony using tissue Doppler echocardiographic imaging (TDI). Two-dimensional and Doppler echocardiography were used for quantification of MR. Statistical analyses were performed using unpaired t test, Mann-Whitney U test, correlation and logistic regression analyses. Thirty-seven patients (39%) had significant DYS Sep-Lat Sys and 25 patients (26%) had DYS Inter PAP Sys. Patients with DYS Inter PAP Sys had lower basal septum systolic (p=0.007) and late diastolic velocities (p=0.049), greater MR volume (p=0.01), effective regurgitant orifice (ERO) (p=0.01), and E/A ratios (p=0.03) than the patients without DYS Inter PAP Sys. Fifty-five patients with narrow QRS intervals were also evaluated for DYS Inter PAP Sys. Patients with DYS Inter PAP Sys and narrow QRS had lower basal septum TDI peak systolic velocities (p=0.038), higher MR volume (p=0.03) and ERO (p=0.03). Logistic regression analysis revealed that NYHA Class III-IV (OR=6.4, 95% CI: 1.1-37.1, p=0.038) and DYS Inter PAP Sys (OR=9.5, 95% CI: 1.17-75.78, p=0.034) were the independent predictors of functional MR >20 ml. Papillary muscle systolic dyssynchrony is common and correlated with functional MR in non-ischemic DCM patients with sinus rhythm and narrow QRS. Papillary muscle systolic dyssynchrony may help predict patients who will benefit from cardiac resynchronization therapy.


Validation study of neurotrophin-3-releasing chitosan facilitation of neural tissue generation in the severely injured adult rat spinal cord.
It was previously reported that a tube holding chitosan carriers loaded with neurotrophin-3 (NT-3), after insertion into a 5 mm long transection gap in the adult rat spinal cord, triggered de novo neural tissue generation and functional recovery. Here, we report an effort to validate these findings using stringent blinding methodologies, which are crucial for robustness in reproducing biomedical studies. Radio frequency identification (RFID) chips were utilized to label rats that were randomly assigned into three experimental groups: transection with chitosan-NT-3 implant (C-NT3), transection only (T-controls), and laminectomy only (S-controls), blinding the experimenters to the treatments. Three months after surgery, animals only known by their RFID were functionally, electrophysiologically, and anatomically assessed. The data were then collected into the proper groups and statistically analyzed. Neural tissue with nestin-, Tuj1-, and NeuN-positive cells was found bridging the transection gap in C-NT3 rats, but not in T-controls. Motor- and somatosensory-evoked potentials were detected in C-NT3 rats and S-controls, but not in T-controls. Hind limb movement was significantly better in C-NT3 rats compared with T-controls. Our validation study indicates that C-NT3 implants facilitate neural tissue generation, at least in part, by eliciting endogenous neurogenesis. Our data support the use of C-NT3 implants for tissue remodeling in the injured spinal cord.


Design and control of glycerol-tert-butyl alcohol etherification process.
Design, economics, and plantwide control of a glycerol-tert-butyl alcohol (TBA) etherification plant are presented. The reaction takes place in liquid phase, in a plug flow reactor, using Amberlyst 15 as a catalyst. The products' separation is achieved by two distillation columns where high-purity ethers are obtained and a section involving extractive distillation with 1,4-butanediol as solvent, which separates TBA from the TBA/water azeotrope. Details of design performed in AspenPlus and an economic evaluation of the process are given. Three plantwide control structures are examined using a mass balance model of the plant. The preferred control structure fixes the fresh glycerol flow rate and the ratio glycerol + monoether : TBA at reactor-inlet. The stability and robustness in the operation are checked by rigorous dynamic simulation in AspenDynamics.


Risk perceptions and trust following the 2010 and 2011 Icelandic volcanic ash crises.
Eruptions at the Icelandic volcanoes of Eyjafjallajökull (2010) and Grimsvötn (2011) produced plumes of ash posing hazards to air traffic over northern Europe. In imposing restrictions on air traffic, regulators needed to balance the dangers of accidents or aircraft damage against the cost and inconvenience to travelers and industry. Two surveys examined how members of the public viewed the necessity of the imposed restrictions and their trust in different agencies as estimators of the level of risk. Study 1 was conducted with 213 British citizens (112 males, 101 females), who completed questionnaires while waiting for flights at London City Airport during May 2012. Study 2 involved an online survey of 301 Icelandic citizens (172 males, 127 females, 2 undeclared gender) during April 2012. In both samples, there was general support for the air traffic restrictions, especially among those who gave higher estimates of the likelihood of an air accident or mishap having otherwise happened. However, in both countries, the (minority of) respondents who had personally experienced travel disruption were less convinced that these restrictions were all necessary. Scientists, the International Civil Aviation Organization, and (in Iceland) the Icelandic Department of Civil Protection were all highly trusted, and seen as erring on the side of caution in their risk estimates. Airlines were seen as more likely to underestimate any risk. We conclude that perceptions of the balance between risk and caution in judgments under uncertainty are influenced by one's own motives and those attributed to others.


Electrical pacemaker mechanisms of pancreatic islet cells.
Glucose, the major physiological stimulus for insulin secretion, induces a periodic bursting pattern of Ca2+ action potentials that are thought to mediate the uptake of Ca2+ into the intracellular pool of free Ca2+, which controls the rate of insulin release. Evidence is reviewed that shows that the voltage-dependent Ca2+ spikes are driven by a slow, voltage-dependent plateau depolarization that may also be caused by Ca2+ influx. Current evidence suggests that this plateau conductance is periodically terminated in turn by a pacemaker current through membrane K+ channels that are activated by intracellular free Ca2+. The control of electrical activity by different modulators of insulin release may involve interactions with this system at several points, including changes of the sensitivity of K+ channels to intracellular Ca2+ and to changes of intracellular Ca2+ buffering capacity.


Prediction of Low and Very Low Bone Mineral Density Among Adult Survivors of Childhood Cancer.
To develop and validate prediction models for low and very low bone mineral density (BMD) on the basis of clinical and treatment characteristics that identify adult survivors of childhood cancer who require screening by dual-energy x-ray absorptiometry. White survivors of childhood cancer (n = 2,032; median attained age, 29.3 years [range, 18.1 to 40.9 years]) enrolled in the St Jude Lifetime Cohort (SJLIFE; development) and survivors treated at the Erasmus Medical Center (validation) in the Netherlands (n = 403; median age, 24.2 years [range, 18.0 to 40.9 years]) were evaluated with dual-energy x-ray absorptiometry to determine lumbar spine BMD and total-body BMD. Low and very low BMD were defined as lumbar spine BMD and/or total-body BMD z scores of -1 or lower or -2 or lower, respectively. Multivariable logistic regression was used to build prediction models; performance was assessed using receiver operating characteristic curves. Diagnostic values were calculated at different probabilities. Low BMD was present in 51% and 45% of SJLIFE and Dutch participants, respectively, and very low BMD was present in 20% and 10%, respectively. The model for low BMD included male sex (odds ratio [OR], 3.07), height (OR, 0.95), weight (OR, 0.98), attained age (OR, 0.97), current smoking status (OR, 1.48), and cranial irradiation (OR, 2.11). Areas under the curve were 0.72 (95% CI, 0.70 to 0.75) in the SJLIFE cohort and 0.69 (95% CI, 0.64 to 0.75) in the Dutch cohort. The sum of the sensitivity (69.0%) and specificity (64.0%) was maximal at the predicted probability of 50%. The model for very low BMD included male sex (OR, 3.28), height (OR, 0.95), weight (OR, 0.97), attained age (OR, 0.98), cranial irradiation (OR, 2.07), and abdominal irradiation (OR, 1.61), yielding areas under the curve of 0.76 (95% CI, 0.73 to 0.78; SJLIFE cohort) and 0.75 (95% CI, 0.67 to 0.83; Dutch cohort). Validated prediction models for low and very low BMD, using easily measured patient and treatment characteristics, correctly identified BMD status in most white adult survivors through age 40 years.


Coronal and root caries in the older Iowans: 9- to 11-year incidence.
This study reports on coronal and root caries incidence in elderly Iowans between 1987 and 1998. The sample consisted of 74 survivors of the Iowa 65+ Oral Health Study cohort who received oral examinations both in 1987 and between 1996-98. The study found that average untreated coronal and root increments were 0.96 and 0.69 surfaces, filled increments were 18.22 and 1.28 surfaces, and combined increments were 18.3 and 1.27 surfaces, respectively. Therefore, annualized untreated coronal and root increments were 0.10 and 0.07 surfaces, filled were 1.80 and 0.13 surfaces, and combined increments were 1.81 and 0.12 surfaces, respectively. The annualized attack rates were 2.13 for coronal and 0.80 for root caries. Approximately 93% of the subjects developed some new coronal and 43% some new root caries increment. Dental caries still constitute a significant problem since a large proportion of the elderly developed caries during this period. This suggests a need for improved preventive and treatment strategies for this aged population.


Ab initio solution and refinement of two high-potential iron protein structures at atomic resolution.
The crystal structure of the reduced high-potential iron protein (HiPIP) from Chromatium vinosum has been redetermined in a new orthorhombic crystal modification, and the structure of its H42Q mutant has been determined in orthorhombic (H42Q-1) and cubic (H42Q-2) modifications. The first two were solved by ab initio direct methods using data collected to atomic resolution (1.20 and 0. 93 A, respectively). The recombinant wild type (rc-WT) with two HiPIP molecules in the asymmetric unit has 1264 protein atoms and 335 solvent sites, and is the second largest structure reported so far that has been solved by pure direct methods. The solutions were obtained in a fully automated way and included more than 80% of the protein atoms. Restrained anisotropic refinement for rc-WT and H42Q-1 converged to R(1) = summation operator||F(o)| - |F(c)|| / summation operator|F(o)| of 12.0 and 13.6%, respectively [data with I > 2sigma(I)], and 12.8 and 15.5% (all data). H42Q-2 contains two molecules in the asymmetric unit and diffracted only to 2.6 A. In both molecules of rc-WT and in the single unique molecule of H42Q-1 the [Fe(4)S(4)](2+) cluster dimensions are very similar and show a characteristic tetragonal distortion with four short Fe-S bonds along four approximately parallel cube edges, and eight long Fe-S bonds. The unique protein molecules in H42Q-2 and rc-WT are also very similar in other respects, except for the hydrogen bonding around the mutated residue that is at the surface of the protein, supporting the hypothesis that the difference in redox potentials at lower pH values is caused primarily by differences in the charge distribution near the surface of the protein rather than by structural differences in the cluster region.


Freezing the Bioactive Conformation to Boost Potency: The Identification of BAY 85-8501, a Selective and Potent Inhibitor of Human Neutrophil Elastase for Pulmonary Diseases.
Human neutrophil elastase (HNE) is a key protease for matrix degradation. High HNE activity is observed in inflammatory diseases. Accordingly, HNE is a potential target for the treatment of pulmonary diseases such as chronic obstructive pulmonary disease (COPD), acute lung injury (ALI), acute respiratory distress syndrome (ARDS), bronchiectasis (BE), and pulmonary hypertension (PH). HNE inhibitors should reestablish the protease-anti-protease balance. By means of medicinal chemistry a novel dihydropyrimidinone lead-structure class was identified. Further chemical optimization yielded orally active compounds with favorable pharmacokinetics such as the chemical probe BAY-678. While maintaining outstanding target selectivity, picomolar potency was achieved by locking the bioactive conformation of these inhibitors with a strategically positioned methyl sulfone substituent. An induced-fit binding mode allowed tight interactions with the S2 and S1 pockets of HNE. BAY 85-8501 ((4S)-4-[4-cyano-2-(methylsulfonyl)phenyl]-3,6-dimethyl-2-oxo-1-[3-(trifluoromethyl)phenyl]-1,2,3,4-tetrahydropyrimidine-5-carbonitrile) was shown to be efficacious in a rodent animal model related to ALI. BAY 85-8501 is currently being tested in clinical studies for the treatment of pulmonary diseases.


Intrinsic direct repeats generate consistent post-transcriptional gene silencing in tobacco.
It is well documented that transgenes with inverted repeats can efficiently trigger post-transcriptional gene silencing (PTGS), presumably via a double stranded RNA induced by complementary sequences in their transcripts. We show here that transgenes with intrinsic direct repeats can also induce PTGS at a very high frequency (80-100%). A transgene with three or four repeats induced PTGS in almost 100% of the primary transformants, regardless of whether a strong (enhanced 35S promoter) or a relatively weak (chlorophyll a/b binding protein promoter) promoter was used. The PTGS induced by three or four repeats is consistently inherited in subsequent generations, and can inactivate homologous genes in trans. Based on the high frequency and consistent heritability, we propose that the intrinsic direct repeat within a transgene may act as a primary determinant of PTGS referred to as direct repeat-induced PTGS (driPTGS). Silencing occurred in all five genes, in this and two previous reports, suggesting that driPTGS might be a universal gene silencing mechanism both in dicotyledonous tobacco plants and monocotyledonous rice cells. In addition, driPTGS may help dissect the gene silencing mechanism and generate silenced phenotypes useful for research and plant biotechnology products.


[On the mechanism of antimotion sickness effects of mexidol].
The path-clamp method used within the whole-cell configuration in experiments with convoluted medullar oblongata sections obtained from white mongrel male rats aged 13 to 17 days evidenced that 5 mV of mexidol caused 96 +/- 2% inhibition of the excitation postsynaptic current in neurons of the medial vestibular nucleus generated by the depolarization step of 10 mV (holding potential = -70 my). This means that the antimotion sickness effect of mexidol has its origin in the ion mechanisms with involvement of the glutamate- and GABAergic components, primarily inhibition of ion currents through channels of the NMDA-receptor complex.


Solid-state carbon-13 nuclear magnetic resonance of humic acids at high magnetic field strengths.
Use of solid-state 13C nuclear magnetic resonance (NMR) spectroscopy has become commonplace in studies of humic substances in soils and sediments, but when modern high-field spectrometers are employed, care must be taken to ensure that the data obtained accurately reflect the chemical composition of these complex materials in environmental systems. In an effort to evaluate the quality of solid-state 13C NMR spectra obtained with modern high-field spectrometers, we conducted a series of experiments to examine spectra of various humic acids taken under a variety of conditions. We evaluate conditions for obtaining semiquantitative cross polarization magic angle spinning (CPMAS) 13C NMR spectra of humic acids at high magnetic field and spinning frequency. We examine the cross polarization (CP) dynamics under both traditional and ramp CP conditions on Cedar Creek humic acid. Fitted equilibrium intensities from these CP dynamic studies compare to within 3.4% of the intensities determined from a Bloch decay spectrum of the same sample. With a 1-ms contact time, ramp CP and traditional CP spectra were acquired on this sample and were found to compare to within 5.4% of the Bloch decay spectrum; however, the signal-to-noise ratio per hour of data acquisition was found to double under ramp CP conditions. These results demonstrate the power of applying modern solid-state NMR techniques at high magnetic field strengths. With these techniques, high-quality, semiquantitative spectra can be quickly produced, allowing the application of solid-state NMR techniques to more environmentally relevant samples, especially those where the quantity is limited.


Glomus tumour: a retrospective review of 15 years experience in a single institution.
Glomus tumours (GT), neoplasms of the glomus body comprise 4.5% of upper limb tumours. Seventy-five per cent of these occur in the hand, and most are subungual (50%). We performed a literature review and retrospective search of histopathologically confirmed GT seen from 1995 to 2009. Fifteen patients were identified, with an average age of 49.6 years. Eight were in the hand, 2 in the upper limb, 2 lower limb and 3 in the ear. Eighty-six per cent presented with pain and 50% underwent radiological investigation. Most diagnoses followed biopsy findings. Surgical excision resulted in a recurrence rate of 13%. The average time to diagnosis was 3.3 years. The average duration of symptoms was 7 to 11 years with an average of 2 to 3 consultations prediagnosis. MRI remains the most useful imaging modality (82 to 90% sensitivity). Excision biopsy is the most common treatment. Greater awareness is needed for quicker diagnosis. Glomus Tumour, Presentation, Imaging, Recurrence.


In situ imaging and control of layer-by-layer femtosecond laser thinning of graphene.
Although existing methods (chemical vapor deposition, mechanical exfoliation, etc.) are available to produce graphene, the lack of thickness control limits further graphene applications. In this study, we demonstrate an approach to precisely thin graphene films to a specific thickness using femtosecond (fs) laser raster scanning. By using appropriate laser fluence and scanning times, graphene thinning with an atomic layer precision, namely layer-by-layer graphene removal, has been realized. The fs laser used was configured in a four-wave mixing (FWM) system which can be used to distinguish graphene layer thickness and count the number of layers using the linear relationship between the FWM signal intensity and the graphene thickness. Furthermore, FWM imaging has been successfully applied to achieve in situ, real-time monitoring of the fs laser graphene thinning process. This method can not only realize the large-scale thinning of graphene with atomic layer precision, but also provide in situ, rapid imaging capability of graphene for an accurate assessment of the number of layers.


Intraarticular Injection of a Cross-Linked Sodium Hyaluronate Combined with Triamcinolone Hexacetonide (Cingal) to Provide Symptomatic Relief of Osteoarthritis of the Knee: A Randomized, Double-Blind, Placebo-Controlled Multicenter Clinical Trial.
To evaluate the efficacy and safety of an intraarticular injection of Cingal (Anika Therapeutics, Inc., Bedford, MA) compared with Monovisc (Anika Therapeutics, Inc., Bedford, MA) or saline for the treatment of knee osteoarthritis. This multicenter, double-blind, saline-controlled clinical trial randomized subjects with knee osteoarthritis (Kellgren-Lawrence grades I-III) to a single injection of Cingal (4 mL, 88 mg hyaluronic acid [HA] plus 18 mg triamcinolone hexacetonide [TH]), Monovisc (4 mL, 88 mg HA), or saline (4 mL, 0.9%). The primary efficacy outcome was change in WOMAC (Western Ontario and McMaster Universities Arthritis Index) Pain Score through 12 weeks with Cingal versus saline. Secondary outcomes included Patient and Evaluator Global Assessments, OMERACT-OARSI Responder index, and WOMAC Total, Stiffness, and Physical Function scores through 26 weeks. A total of 368 patients were treated (Cingal, n = 149; Monovisc, n = 150; saline, n = 69). Cingal improvement from baseline was significantly greater than saline through 12 weeks ( P = 0.0099) and 26 weeks ( P = 0.0072). WOMAC Pain was reduced by 70% at 12 weeks and by 72% at 26 weeks with Cingal. Significant improvements were found in most secondary endpoints for pain and function at most time points through 26 weeks. At 1 and 3 weeks, Cingal was significantly better than Monovisc for most endpoints; Cingal and Monovisc were similar from 6 weeks through 26 weeks. A low incidence of related adverse events was reported. Cingal provides immediate and long-term relief of osteoarthritis-related pain, stiffness, and function, significant through 26 weeks compared to saline. Cingal had similar immediate advantages compared with HA alone, while showing benefit comparable to HA at 6 weeks and beyond.