CYP2D6 genotype and induction of intestinal drug transporters by rifampin predict presystemic clearance of carvedilol in healthy subjects.
Clinical trials have indicated that the combined beta- and alpha-adrenergic receptor blocker carvedilol improves the survival rate in patients with advanced chronic heart failure. The objective of our study was the identification and quantification of factors that modulate steady-state serum concentrations of carvedilol and its enantiomers and that may influence therapeutic efficacy and safety. The influence of genetic variants of cytochrome P450 (CYP) 2D6 and CYP2C9 and of transporter proteins (P-glycoprotein, multidrug resistance protein 2 [MRP2]) on the disposition of carvedilol and its enantiomers after intravenous (5 mg) and long-term oral administration (25 mg for 7 days) was assessed in 12 healthy subjects. The intestinal expression of P-glycoprotein and MRP2 was analyzed by quantitative real-time polymerase chain reaction and immunohistochemical techniques. The area under the serum concentration-time curve (AUC) values of carvedilol were significantly (P <.05) increased in 6 subjects with CYP2D6 deficiency, with effects being more pronounced for R(+)-carvedilol (230 +/- 72.6 ng. h/mL versus 93.9 +/- 64.6 ng. h/mL in extensive metabolizers) than for S(-)-carvedilol (62.9 +/- 21.1 ng. h/mL versus 32.7 +/- 14.5 ng. h/mL). The AUC and fecal excretion of intravenous carvedilol were correlated with the intestinal expression of MDR1 messenger ribonucleic acid (mRNA) (r = -0.67, P =.001; r = 0.83, P =.002) and MRP2 mRNA (r = -0.74, P <.001; r = 0.70, P =.025). Furthermore, we measured the disposition of long-term oral carvedilol after comedication of the pregnane X receptor ligand rifampin (INN, rifampicin) (600 mg, 9 days), which up-regulates both P-glycoprotein and MRP2 but not CYP2D6. Rifampin decreased the AUC of carvedilol to an extent independent of the CYP2D6 genotype (poor metabolizers, 341 +/- 147 ng. h/mL versus 126 +/- 41.7 ng. h/mL; extensive metabolizers, 173 +/- 102 ng. h/mL versus 74 +/- 41.4 ng. h/mL; both P <.05). The AUC was significantly correlated with intestinal expression of MDR1 mRNA (r = -0.671, P =.001) and MRP2 mRNA (r = -0.595, P <.006). Variable plasma concentrations of carvedilol during long-term administration are predicted by CYP2D6 genotype and intestinal expression of P-glycoprotein and MRP2.


Successful Systemic Thrombolysis in an Adolescent With Acute Ischemic Stroke.
Current treatment guidelines for acute ischemic stroke do not recommend thrombolytic therapy in children and adolescents as data are still very scarce. We report the case of a 15-year-old boy who suddenly developed severe left-sided weakness and speech difficulty while stooling. Upon arrival at our Emergency Department, the National Institute of Health Stroke Scale (NIHSS) score was 18. Urgent neurovascular ultrasound showed a distal occlusion of the right internal carotid artery and occlusion at the origin of the middle cerebral artery (MCA) and the anterior cerebral artery. He was treated 2 hours after symptom onset with intravenous recombinant tissue plasminogen activator without any complication. At the end of thrombolysis, a complete recanalization was shown by transcranial Doppler sonography, although a brain magnetic resonance imaging disclosed an acute right middle cerebral artery stroke. At discharge, the boy had mild weakness on his left leg and slight left facial palsy: the NIHSS score was 2. To our knowledge, this is the first intravenous thrombolytic treatment ever reported in an adolescent in Italy. Despite the lack of evidence regarding the safety and the efficacy of recombinant tissue plasminogen activator in pediatric stroke, this treatment option should be considered, especially in adolescents presenting within 3 hours from symptom onset in centers with consolidated experience in adult thrombolysis.


c-Myc-activated USP2-AS1 suppresses senescence and promotes tumor progression via stabilization of E2F1 mRNA.
The c-Myc oncoprotein plays a prominent role in cancer initiation, progression, and maintenance. Long noncoding RNAs (lncRNAs) are recently emerging as critical regulators of the c-Myc signaling pathway. Here, we report the lncRNA USP2-AS1 as a direct transcriptional target of c-Myc. Functionally, USP2-AS1 inhibits cellular senescence and acts as an oncogenic molecule by inducing E2F1 expression. Mechanistically, USP2-AS1 associates with the RNA-binding protein G3BP1 and facilitates the interaction of G3BP1 to E2F1 3'-untranslated region, thereby leading to the stabilization of E2F1 messenger RNA. Furthermore, USP2-AS1 is shown as a mediator of the oncogenic function of c-Myc via the regulation of E2F1. Together, these findings suggest that USP2-AS1 is a negative regulator of cellular senescence and also implicates USP2-AS1 as an important player in mediating c-Myc function.


Interactions between the nucleus and cytoplasmic organelles during the cell cycle of Euglena gracilis in synchronized cultures. IV. An aggregate form of chloroplasts in association with the nucleus appearing prior to chloroplast division.
Cells of Euglena gracilis were synchronized by applying a 14-h light:10-h dark regimen under photoautotrophic conditions and a 10-h light:14-h dark regimen under photoorganotrophic conditions. At a stage just prior to chloroplast division in the cell cycle of these synchronized cultures, chloroplasts temporarily gathered in the posterior part of the cell and were connected to each other by many bridges. Part of the chloroplast aggregate surrounded about half of the nuclear surface, making connections or close contacts at many sites. A chromosome was always attached to the inner membrane of the nuclear envelope at the site of association with the chloroplast. The nucleoids in these aggregate chloroplasts, examined by staining with 4',6-diamidino-2-phenylindole, a DNA fluorochrome, showed profiles of strings or strands with branchings, under photoorganotrophic conditions at least, and some parts of the branchings came close to the site of association with the nucleus. The association between the chloroplast aggregate and the nucleus was also observed in Euglena cells placed in continuous darkness after synchronization under photoorganotrophic conditions, suggesting that these organellar associations are related to the Euglena cell cycle but are not the result of light:dark alternations used for cell synchronization.


Automatic quantification of myocardium and pericardial fat from coronary computed tomography angiography: a multicenter study.
To develop a deep learning-based method for simultaneous myocardium and pericardial fat quantification from coronary computed tomography angiography (CCTA) for the diagnosis and treatment of cardiovascular disease (CVD). We retrospectively identified CCTA data obtained between May 2008 and July 2018 in a multicenter (six centers) CVD study. The proposed method was evaluated on 422 patients' data by two studies. The first overall study involves training model on CVD patients and testing on non-CVD patients, as well as training on non-CVD patients and testing on CVD patients. The second study was performed using the leave-center-out approach. The method performance was evaluated using Dice similarity coefficient (DSC), Jaccard index (JAC), 95% Hausdorff distance (HD95), mean surface distance (MSD), residual mean square distance (RMSD), and the center of mass distance (CMD). The robustness of the proposed method was tested using the nonparametric Kruskal-Wallis test and post hoc test to assess the equality of distribution of DSC values among different tests. The automatic segmentation achieved a strong correlation with contour (ICC and R > 0.97, p value < 0.001 throughout all tests). The accuracy of the proposed method remained high through all the tests, with the median DSC higher than 0.88 for pericardial fat and 0.96 for myocardium. The proposed method also resulted in mean MSD, RMSD, HD95, and CMD of less than 1.36 mm for pericardial fat and 1.00 mm for myocardium. The proposed deep learning-based segmentation method enables accurate simultaneous quantification of myocardium and pericardial fat in a multicenter study. • Deep learning-based myocardium and pericardial fat segmentation method tested on 422 patients' coronary computed tomography angiography in a multicenter study. • The proposed method provides segmentations with high volumetric accuracy (ICC and R > 0.97, p value < 0.001) and similar shape as manual annotation by experienced radiologists (median Dice similarity coefficient ≥ 0.88 for pericardial fat and 0.96 for myocardium).


Protein NMR in biomedical research.
Nuclear magnetic resonance (NMR) spectroscopy is a versatile biophysical technique with wide applicability in drug discovery research, particularly for the detection and characterization of molecular interactions. This review highlights in a comprehensive manner the aspects of biomolecular NMR which are most beneficial for pharmaceutical research and presents them as contributions to the different stages of a drug discovery program: target selection, assay development, lead generation and lead optimization. Emphasis is put on the concept of the particular NMR application, rather than on technical details, and on recent examples. Finally, an appendix of frequently asked questions is given.


Integrated vermi-pisciculture--an alternative option for recycling of solid municipal waste in rural India.
Vermicomposts as a biofertilizer can be a great option for pond manuring as they never cause any long term harm to the soil like chemical fertilizer. In this study vegetable and horticulture waste was used as an important media for vermiculture. Three separate cemented tanks (6 m(3) each) were used in the system as control tank, vermicompost fertilized tank and inorganic fertilizer manured tank. Monoculture of fish was carried out with cat fish, Clarias batrachus. The produced earthworms were used as fish feed. Regular monitoring of water parameter was conducted in three different ponds. Specifically, the algal biomass variation was quite helpful in analyzing the behavior of the ponds. NPK value of soil samples were analyzed intermittently to know the eutrophication level. Despite the hot summer temperature in northern part of India, which is not ideal for fish growth, we have recorded an encouraging growth performance in organic manured pond along with inorganic fertilizer treated and control pond. Among eutrophicated pond, the fish biomass from vermicompost fed pond showed an increasing trend compared to inorganic fertilizer treated pond. Water retention capacity of vermicompost pond soil was better in comparison to other ponds. Result shows that the low cost model by integrating two production system vermiculture and pisciculture could be a commercially and environmentally viable option.


Tissue distribution and function of the Aryl hydrocarbon receptor repressor (AhRR) in C57BL/6 and Aryl hydrocarbon receptor deficient mice.
The Aryl hydrocarbon receptor repressor (AhRR) is a new member of bHLH-PAS proteins which is important in the regulation of cell growth and differentiation. The AhRR shares structural similarities with Aryl hydrocarbon receptor (AhR) and AhR nuclear translocator (ARNT). The AhRR is thought to be involved in transcriptional control of AhR-regulated genes by sequestering ARNT. Most of the knowledge of regulation and function of the AhRR is from studies in cell lines. Here, we report the tissue distribution of AhRR in AhR deficient and wild type C57BL/6 mice. In addition, the inducibility of the AhRR and Cytochrome P450 (CYP) 1A1 in response to benzo(a)pyrene (B(a)P) (10 mg/kg bw i.p.) was investigated. The results show that the AhRR mRNA expression pattern in untreated C57BL/6 mice varies across tissues with high levels in hearts and brains. In other tissues, AhRR mRNA expression was low. In contrast to wild-type animals, the tissue levels in AhR-/- mice were about two to three orders of magnitude lower. Treatment of wild-type animals with B(a)P resulted in an induced AhRR expression in liver, spleen, lung and ovary. No significant induction of AhRR mRNA was found in brain and heart tissues, which have a constitutively high level of AhRR expression. Simultaneous measurements of CYP1A1 and AhRR mRNA expression do not strongly support the view that the AhRR tissue pattern triggers the tissue specific responsiveness of AhR-regulated genes to B(a)P treatment.


MR imaging features of nongynaecologic cystic lesions of the pelvis.
The purpose of our article is to review the magnetic resonance imaging (MRI) features of nongynaecologic cystic lesions of the pelvis. The rising use of MRI for pelvic exploration will result in an increase in incidental detection of pelvic cystic cysts. Pelvic cysts of non gynecologic origin are less frequent than gynecologic cysts. However, they account for a wide range of abnormalities, and radiologists must be aware of their features and characteristics.


Cytoskeleton alterations induced by Geodia corticostylifera depsipeptides in breast cancer cells.
Crude extracts of the marine sponge Geodia corticostylifera from Brazilian Coast have previously shown antibacterial, antifungal, cytotoxic, haemolytic and neurotoxic activities. The present work describes the isolation of the cyclic peptides geodiamolides A, B, H and I (1-4) from G. corticostylifera and their anti-proliferative effects against sea urchin eggs and human breast cancer cell lineages. Its structure-activity relationship is discussed as well. In an initial series of experiments these peptides inhibited the first cleavage of sea urchin eggs (Lytechinus variegatus). Duplication of nuclei without complete egg cell division indicated the mechanism of action might be related to microfilament disruption. Further studies showed that the geodiamolides have anti-proliferative activity against human breast cancer cell lines (T47D and MCF7). Using fluorescence techniques and confocal microscopy, we found evidence that the geodiamolides A, B, H and I act by disorganizing actin filaments of T47D and MCF7 cancer cells, in a way similar to other depsipeptides (such as jaspamide 5 and dolastatins), keeping the normal microtubule organization. Normal cells lines (primary culture human fibroblasts and BRL3A rat liver epithelial cells) were not affected by the treatment as tumor cells were, thus indicating the biomedical potential of these compounds.


Unstable, heterogeneous physical states of nuclear bodies during the post-hepatectomy dedifferentiation-replication-redifferentiation sequence.
Nuclei are shown to undergo generalized changes of state during regeneration, as reflected by changes in equilibrium density of sucrose density gradients. The time-course of these phenomena suggests an association with changes occurring after S-phase and mitosis. Such changes are no longer observable in extensively purified nuclei (washed, polyamine-stabilized nuclei), but are apparent only in nuclei with minimal purification, rapidly isolated by centrifugation from liver homogenates. This effect implies that there exists a serious risk of selective loss of some nuclear types during standard purification procedures that use 2.1-2.4 M sucrose cushions. Although these nuclear states may largely reflect the metabolic state of the cells of origin, evidence is presented to suggest that loss and redistribution of macromolecules during handling procedures causes distortion of in vivo properties.


Fermi surface topology and the upper critical field in two-band superconductors: application to MgB2.
Recent measurements of the anisotropy of the upper critical field B(c2) on MgB2 single crystals have shown a puzzling strong temperature dependence. Here, we present a calculation of the upper critical field based on a detailed modeling of band structure calculations that takes into account both the unusual Fermi surface topology and the two gap nature of the superconducting order parameter. Our results show that the strong temperature dependence of the B(c2) anisotropy can be understood as an interplay of the dominating gap on the sigma band, which possesses a small c-axis component of the Fermi velocity, with the induced superconductivity on the pi-band possessing a large c-axis component of the Fermi velocity. We provide analytic formulas for the anisotropy ratio at T=0 and T=T(c) and quantitatively predict the distortion of the vortex lattice based on our calculations.


ECEL1 mutation implicates impaired axonal arborization of motor nerves in the pathogenesis of distal arthrogryposis.
The membrane-bound metalloprotease endothelin-converting enzyme-like 1 (ECEL1) has been newly identified as a causal gene of a specific type of distal arthrogryposis (DA). In contrast to most causal genes of DA, ECEL1 is predominantly expressed in neuronal cells, suggesting a unique neurogenic pathogenesis in a subset of DA patients with ECEL1 mutation. The present study analyzed developmental motor innervation and neuromuscular junction formation in limbs of the rodent homologue damage-induced neuronal endopeptidase (DINE)-deficient mouse. Whole-mount immunostaining was performed in DINE-deficient limbs expressing motoneuron-specific GFP to visualize motor innervation throughout the limb. Although DINE-deficient motor nerves displayed normal trajectory patterns from the spinal cord to skeletal muscles, they indicated impaired axonal arborization in skeletal muscles in the forelimbs and hindlimbs. Systematic examination of motor innervation in over 10 different hindlimb muscles provided evidence that DINE gene disruption leads to insufficient arborization of motor nerves after arriving at the skeletal muscle. Interestingly, the axonal arborization defect in foot muscles appeared more severe than in other hindlimb muscles, which was partially consistent with the proximal-distal phenotypic discordance observed in DA patients. Additionally, the number of innervated neuromuscular junction was significantly reduced in the severely affected DINE-deficient muscle. Furthermore, we generated a DINE knock-in (KI) mouse model with a pathogenic mutation, which was recently identified in DA patients. Axonal arborization defects were clearly detected in motor nerves of the DINE KI limb, which was identical to the DINE-deficient limb. Given that the encoded sequences, as well as ECEL1 and DINE expression profiles, are highly conserved between mouse and human, abnormal arborization of motor axons and subsequent failure of NMJ formation could be a primary cause of DA with ECEL1 mutation.


Release of histamine from leucocytes and its determinants in vitro in relation to bronchial responsiveness to inhaled histamine and exercise in vivo.
The hypothesis studied is that increased responsiveness in asthma is not limited to the airways. Forty asthmatic children were analysed for their bronchial responsiveness (BR) to exercise. Twenty patients revealed bronchial obstruction after exercise while the remainder did not. These observations were compared with the responsiveness of leucocytes, which was determined by their histamine 'releasability'. Twenty healthy children served as controls. Release of histamine induced by calcium ionophore-aided calcium influx was significantly higher in both groups of asthmatics than in the healthy children (P less than 0 X 005). Similar findings were obtained by induction of microtubule aggregation due to deuterium oxide (D2O). The S-shaped dose-response relationship with D2O was shifted to the left in the patients with BR to exercise compared to patients without (P less than 0 X 025). The slope was increased in both patient groups compared with the healthy children (P less than 0 X 01). It is concluded that the mean 'releasability' of histamine release due to both stimulants correlated well (P less than 0 X 01). This suggests that the 'releasability' is determined by the responsiveness of the microtubules. This may also apply to allergen-induced histamine release, as was revealed from studies with anti-IgE. The differences in histamine release found in relation to BR due to exercise were also present if the patients were divided according to BR due to histamine. A significant relationship existed between the degree of BR to histamine and the responsiveness of the microtubules (P less than 0 X 02).


Post-synaptic activity evoked in the rostral ventrolateral medullary neurones by stimulation of the defence areas of hypothalamus and midbrain in the rat.
Neurones in the rostral ventrolateral medulla (RVLM) of the rat were recorded intracellularly (n = 30) and extracellularly (n = 91) in vivo. 91% of them had spontaneous activity with frequencies of 1.1-29.9 Hz. Onset latencies of the excitation induced by the stimulation of the hypothalamic and midbrain defence areas ranged from 1.5 to 44 ms and 2 to 60 ms respectively. There was no statistical difference between two groups. Excitation followed by inhibition or inhibition followed by excitation was observed in these processes. Excitatory post-synaptic potentials (EPSPs) were summated by simultaneous stimulation of both sites and the onset latency was changed with the change of stimulus intensity. It is concluded that projections from the defence areas of hypothalamus and midbrain are relayed to RVLM neurones forming excitatory and inhibitory synapses; one mechanism of the effect summation caused by both sites is via EPSPs.


Connecting complex disorders through biology.
Mutations in CTC1, which encodes a key telomere component, have been identified as the cause of Coats plus syndrome. This discovery provides an important pathophysiological link between Coats plus and the clinically related telomere disorders dyskeratosis congenita, Revesz syndrome and Hoyeraal-Hreidarsson syndrome.


The effects of combined early oral health care and early mobilisation on the incidence of stroke-associated pneumonia.
To investigate the effects of combined early oral healthcare and early mobilisation on the incidence of stroke-associated pneumonia during hospitalisation of acute stroke patients. In this single-centre, non-blinded, before-and-after cohort study, patients received basic stroke rehabilitation by a multidisciplinary team within 72 h of symptom onset from July to September 2016 and from July to September 2018. Patients were divided into two groups: 1) patients who received combined early oral healthcare and early mobilisation (early intervention group) (n=107), and 2) patients who received usual care (control group) (n=107). The relationship between the stroke-associated pneumonia incidence and prognosis was examined. The early intervention group had a significantly lower incidence of stroke-associated pneumonia than the control group (0.93% vs. 7.48%; P=0.01). Moreover, the early intervention group had a significantly lower proportion of patients who died or required medical care because of recurrent pneumonia at discharge (0.93% vs. 5.6%; P=0.04). In contrast, there were no significant differences between the two groups regarding the Revised Hasegawa's Dementia Scale on day 14 (22.5 vs. 23; P=0.87), Functional Independence Measure on day 14 (112 vs. 116; P=0.06), and rate of total oral diet (Food Intake LEVEL Scale ≥7) at discharge (95.2% vs. 93.5%; P=0.55). Combined early oral healthcare and early mobilisation by a multidisciplinary team significantly decreased the stroke-associated pneumonia incidence within 7 days and reduced the percentage of patients who died or required medical care because of recurrent pneumonia after stroke.


Clobazam higher-evening differential dosing as an add-on therapy in refractory epilepsy.
Clobazam treatment tailored to the timing of patient's seizures may improve seizure control. We aim to describe the safety and efficacy of higher-evening differential dose of clobazam as add-on therapy in patients with night-time/early morning seizures. Differential dosing with higher evening dosing was started based on a high proportion of seizures (>80%) at nighttime (6p.m. to 6a.m.). Differential dosing was defined as providing more than 50% of the total daily dose of clobazam after 6p.m. Twenty-seven patients were treated with clobazam differential dosing as an add-on therapy. The median age was 9.1 years, with 11 (40.7%) females and median of the first follow-up was 2.7 months. Patients with differential dosing tolerated a higher median total clobazam dose of 0.8mg/kg/d at first follow-up, as compared to 0.6mg/kg/d in controls. In differential dose, the median percentage of the total clobazam dose administered in the evening was 66.7%. Differential dose patients exhibited a median seizure reduction of 75% as compared to 50% in controls (p<0.005). Patients with generalized seizures benefited the most from differential dosing with a 77.5% median seizure reduction, as compared to 50% in controls (p=0.017). Higher-evening differential dose of clobazam improved seizure control in patients with predominantly nighttime and early-morning seizures. Chronotherapy tailored to the patients' seizure susceptibility patterns may improve care in epilepsy patients as differential dosing may allow for higher overall treatment doses at times of greatest seizure susceptibility without increased side effects at other times.


Effect of systemic L-DOPA administration on extracellular kynurenate levels in the rat striatum.
Dopaminergic stimulation, caused by direct dopamine receptor agonists or by indirectly acting agents such as d-amphetamine or methylphenidate, causes a functionally significant decrease in the concentration of kynurenate (KYNA), an astrocyte-derived endogenous antagonist of both N-methyl-D-aspartate (NMDA) and alpha7 nicotinic acetylcholine receptors, in the rat brain. The present study was designed to examine if this effect can be duplicated by the systemic administration of dopamine's bioprecursor L-DOPA. Experiments were conducted in unanesthetized rats, and the extracellular levels of KYNA were determined in striatal microdialysate samples. L-DOPA caused a dose-dependent, transient reduction in striatal KYNA, reaching a nadir of -37.5% 1.5 h after the administration of 200 mg/kg of the drug. This effect was abolished in animals with a 6-hydroxydopamine-induced lesion of the nigrostriatal pathway, but was not influenced by a prior striatal quinolinate lesion. These data confirm the dopaminergic control of striatal KYNA formation and suggest that the interactions are mediated by astrocytic dopamine receptors. This modulation of striatal KYNA levels might provide a link between dopaminergic, glutamatergic and cholinergic neurotransmission in the normal and diseased striatum.


Core Entrustable Professional Activities (EPAs) and the Transition from Medical School to Residency: the Postgraduate Year One Resident Perspective.
The Association of American Medical Colleges (AAMC) proposed thirteen core Entrustable Professional Activities (EPAs) that all graduates should be able to perform under indirect supervision upon entering residency. As an underlying premise is that graduates ready to do so will be better prepared to transition to the responsibilities of residency, we explored the relationship between postgraduate year (PGY)-1 residents' self-assessed preparedness to perform core EPAs under indirect supervision at the start of residency with their ease of transition to residency. Using response data to a questionnaire administered in September 2019 to PGY-1 residents who graduated from AAMC core EPA pilot schools, we examined between-group differences and independent associations for each of PGY-1 position type, specialty, and "EPA-preparedness" score (proportion of EPAs the resident reported as prepared to perform under indirect supervision at the start of residency) and ease of transition to residency (from 1 = much harder to 5 = much easier than expected). Of 274 questionnaire respondents (19% of 1438 graduates), 241 (88% of 274) had entered PGY-1 training and completed all questionnaire items of interest. EPA-preparedness score (mean 0.71 [standard deviation 0.26]) correlated with ease of transition (3.1 [0.9]; correlation = .291, p < .001). In linear regression controlling for specialty (among other variables), EPA-preparedness score (β-coefficient 1.08; 95% confidence interval .64-1.52; p < .001) predicted ease of transition to residency. Graduates who felt prepared to perform many of the core EPAs under indirect supervision at the start of PGY-1 training reported an easier-than-expected transition to residency. The online version contains supplementary material available at 10.1007/s40670-021-01370-3.


Contributing factors of crash injury severity at public highway-railroad grade crossings in the U.S.
The Moving Ahead for Progress in the 21st Century (MAP-21) includes a separate program that supports safety improvements to reduce the number of fatalities and injuries at public highway-railroad grade crossings (HRGCs). This study identifies the significant factors affecting crash injury severity at public HRGCs in the United States. Crashes from 2009 through 2013 on 5,528 public HRGCs, extracted from the Federal Railroad Administration database, were used in the analysis. A comprehensive list of risk factors was explored. Examples include predictors related to geographic region of crash, geometry (e.g., area type and pavement marking type), railroad (e.g., warning device type and railroad class), traffic (e.g., train speed and vehicles annual average daily traffic "AADT"), highway user (e.g., driver age and gender), and environment (e.g., lighting and weather conditions). The study used the mixed logit model to better capture the complex highway user behavior at HRGCs. Female highway users were at higher risk of involvement in injuries and fatalities compared to males. Higher train speeds, very old drivers, open areas, concrete road surface types, and railroad equipment striking highway users before crash, were all found to increase the injury likelihood. On the other hand, young and middle-age drivers, non-passing of standing vehicles at HRGCs, industrial areas, and presence of warning bells were found to reduce injuries and fatalities. The mixed logit model succeeded in identifying contributing factors of crash severity at public HRGCs and potential countermeasures to reduce both fatalities and injuries are suggested. It is important to install warning bells at public HRGCs, especially at those with high number of injury and fatality crashes. Enforcement of traffic nearby HRGCs is necessary to prevent vehicles from overtaking of standing vehicles.


Folding of a de novo designed native-like four-helix bundle protein.
The folding of a model native-like dimeric four-helix bundle protein, (alpha(2))(2), was investigated using guanidine hydrochloride, hydrostatic pressure, and low temperature. Unfolding by guanidine hydrochloride followed by circular dichroism and intrinsic fluorescence spectroscopy revealed a highly cooperative transition between the native-like and unfolded states, with free energy of unfolding determined from CD data, DeltaG(unf) = 14.3 +/- 0.8 kcal/mol. However, CD and intrinsic fluorescence data were not superimposable, indicating the presence of an intermediate state during the folding transition. To stabilize the folding intermediate, we used hydrostatic pressure and low temperature. In both cases, dissociation of the dimeric native-like (alpha(2))(2) into folded monomers (alpha(2)) was observed. van't Hoff analysis of the low temperature experiments, assuming a two-state dimer 171-monomer transition, yielded a free energy of dissociation of (alpha(2))(2) of DeltaG(diss) = 11.4 +/- 0.4 kcal/mol, in good agreement with the free energy determined from pressure dissociation experiments (DeltaG(diss) = 10.5 +/- 0.1 kcal/mol). Binding of the hydrophobic fluorescent probe 4,4'-dianilino-1,1'-binaphthyl-5,5'-disulfonic acid (bis-ANS) to the pressure- and cold-dissociated states of (alpha(2))(2) indicated the existence of molten-globule monomers. In conclusion, we demonstrate that the folding pathway of (alpha(2))(2) can be described by a three-state transition including a monomeric molten globule-like state.


Prospective multi-center study of oncologic outcomes of robot-assisted partial nephrectomy for pT1 renal cell carcinoma.
Partial nephrectomy has been increasingly recommended over radical nephrectomy for the management of small renal masses based on improved renal functional outcomes without sacrifice of oncologic effectiveness. Robot-assisted partial nephrectomy (RAPN) has been introduced in an effort to offer another minimally invasive option for nephron-sparing surgery. However, reports of RAPN have been limited to short-term perioperative outcomes. The goal of this study is to report and evaluate the initial oncologic outcomes of RAPN. Utilizing prospectively obtained data on RAPN performed by four surgeons at four separate tertiary care centers, we selected patients with unilateral, localized, non-familial, pathologically-confirmed pT1 renal cell carcinoma and a minimum post-operative follow-up of 12 months. Utilizing prospectively obtained data on RAPN performed by four surgeons at four separate tertiary care centers, we selected patients with unilateral, localized, non-familial, pathologically-confirmed pT1 renal cell carcinoma and a minimum post-operative follow-up of 12 months. Survival analysis (disease-free, cancer-specific, and overall survival) was performed, and Kaplan-Meier curves were generated. RAPN was performed in 124 patients with a median tumor size of 3.0 cm (IQR 2.2-4.2 cm). Median follow-up was 29 months (range 12-46 months). Positive parenchymal surgical margins occurred in two patients (1.6 %), both of whom were recurrence-free at 30 and 34 months after surgery. The three-year Kaplan-Meier estimated disease-free survival was 94.9 %, cancer-specific survival was 99.1%, and overall survival was 97.3 %. In our cohort of patients with small renal carcinomas who were followed for a median of 29 months, recurrence and survival outcomes were similar to those reported for open and laparoscopic partial nephrectomy. Further long-term outcomes will be needed to definitively claim that RAPN is oncologically equivalent to other surgical approaches.


A review of the environmental corrosion, fate and bioavailability of munitions grade depleted uranium.
Depleted uranium (DU) is a by-product of nuclear fuel enrichment and is used in antitank penetrators due to its high density, self-sharpening, and pyrophoric properties. Military activities have left a legacy of DU waste in terrestrial and marine environments, and there have been only limited attempts to clean up affected environments. Ten years ago, very little information was available on the dispersion of DU as penetrators hit their targets or the fate of DU penetrators left behind in environmental systems. However, the marked increase in research since then has improved our knowledge of the environmental impact of firing DU and the factors that control the corrosion of DU and its subsequent migration through the environment. In this paper, the literature is reviewed and consolidated to provide a detailed overview of the current understanding of the environmental behaviour of DU and to highlight areas that need further consideration.


Construction of immunogens for synthetic malaria vaccines.
The immunogenicity of a peptide consisting of eight repeats of the tetrapeptide sequence NANP (Asn-Ala-Asn-Pro) contained in the circumsporozoite protein of Plasmodium falciparum was investigated in mice under different modes of presentation. This peptide was able to produce biologically active antibodies when administered with adjuvant and linked to a protein carrier. However, a (NANP) peptide polymerized by carbodiimide was found to be immunogenic in the absence of protein carrier in H-2b mice. In contrast, the (NANP)8 peptide polymerized by glutaraldehyde was not immunogenic in the same strain. Furthermore, the efficacy of murabutide in saline, as an immunological adjuvant, was compared to the efficacy of Freund's complete adjuvant.


Microcirculation in the macular area of eyes with an idiopathic epiretinal membrane.
To measure the microcirculation in the macula area of eyes before and after the removal of an idiopathic epiretinal membrane (ERM) by vitreous surgery. Tissue blood flow in the macula was examined with the Heidelberg retina flowmeter in 12 eyes with an ERM before and 3, 6, and 12 months after vitrectomy. Blood flow measurements were performed in a 10 x 5 degree area in the macula that was divided into superior and inferior areas. At least three measurements were obtained, and the mean blood flow (MBF) was calculated by scanning laser Doppler flowmetry. The ratio of the MBF in the affected eye to that in the fellow eye was used for statistical analysis. The ratios of the MBF in the affected to the fellow eye in the superior area before and 1, 3, 6, and 12 months after surgery were 0.73, 0.51, 0.77, 0.88, and 1.01, respectively. The ratios of the MBF in the inferior area for the same periods were 0.70, 0.52, 0.75, 0.83, and 0.98, respectively. Eyes with an ERM showed abnormal hemodynamics in the macula that gradually improved after vitrectomy to a status comparable with the MBF of the normal fellow eye.


Complete mitochondrial genomes of two gelechioids, Mesophleps albilinella and Dichomeris ustalella (Lepidoptera: Gelechiidae), with a description of gene rearrangement in Lepidoptera.
We sequenced the entire mitochondrial genome (mitogenome) of two gelechioids, Mesophleps albilinella and Dichomeris ustalella, and compared their genome organization and sequence composition to those of available gelechioid mitogenomes for an enhanced understanding of Gelechioidea genomic characteristics. We compared all available lepidopteran mitogenome arrangements, including that of M. albilinella, which is unique in Gelechioidea, to comprehend the extensiveness and mechanisms of gene rearrangement in Lepidoptera. The genomes of M. albilinella and D. ustalella are 15,274 and 15,410 bp in size, respectively, with the typical sets of mitochondrial (mt) genes. The COI gene begins with CGA (arginine) in all sequenced gelechioids, including M. albilinella and D. ustalella, reinforcing the feature as a synapomorphic trait, at least in the Gelechioidea. Each 353- and 321-bp long A + T-rich region of M. albilinella and D. ustalella contains one (D. ustalella) or two (M. albilinella) tRNA-like structures. The M. albilinella mitogenome has a unique gene arrangement among the Gelechioidea: ARNESF (the underline signifies an inverted gene) at the ND3 and ND5 junction, as opposed to the ARNSEF that is found in ancestral insects. An extensive search of available lepidopteran mitogenomes, including that of M. albilinella, turned up six rearrangements that differ from those of ancestral insects. Most of the rearrangements can be explained by the tandem duplication-random loss model, but inversion, which requires recombination, is also found in two cases, including M. albilinella. Excluding the MIQ rearrangement at the A + T-rich region and ND2 junction, which is found in nearly all Ditrysia, most of the remaining rearrangements found in Lepidoptera appear to be independently derived in that they are automorphic at several taxonomic scales, although current mitogenomic data are limited, particularly for congeneric data.


Generating daily high-resolution and full-coverage XCO2 across China from 2015 to 2020 based on OCO-2 and CAMS data.
China has set a goal to achieve carbon neutrality by 2060, and satellite remote sensing allows for acquiring large-range and high-resolution carbon dioxide (CO2) data, which can aid in achieving this goal. However, satellite-derived column-averaged dry-air mole fraction of CO2 (XCO2) products often suffer from substantial spatial gaps due to the impacts of narrow swath and clouds. Here, this paper generates daily full-coverage XCO2 data at a high spatial resolution of 0.1° over China during 2015-2020, by fusing satellite observations and reanalysis data in a deep neural network (DNN) framework. Specifically, DNN constructs the relationships between Orbiting Carbon Observatory-2 satellite XCO2 retrievals, Copernicus Atmosphere Monitoring Service (CAMS) XCO2 reanalysis data, and environmental factors. Then, daily full-coverage XCO2 data can be generated based on CAMS XCO2 and environmental factors. Results show that a satisfactory performance is reported in multiform validations, with RMSE and R2 of 0.99 ppm and 0.963 in terms of the sample-based cross-validation, respectively. The independent in-situ validation also indicates high consistency (R2 = 0.866 and RMSE = 1.71 ppm) between XCO2 estimates and ground measurements. Based on the generated dataset, spatial and seasonal distributions of XCO2 across China are investigated, and a growth rate of 2.71 ppm/yr is found from 2015 to 2020. This paper generates long time series of full-coverage XCO2 data, which helps promote our understanding of carbon cycling. The dataset is available from https://doi.org/10.5281/zenodo.7793917.


Estimating the prevalence and strength of non-independent predator effects.
Understanding whether multiple predator species have independent effects on shared prey is critical for understanding community dynamics. We describe the prevalence and strength of non-independence between predators by quantifying the prey's risk of predation and the degree to which it deviates from the risk predicted from a null model of independent predator effects. Specifically, we document how frequently non-independent effects occur among ten different multiple predator combinations with mayfly larvae as prey. These predator combinations vary both predator density and predator species richness. Overall, the predator effects were non-independent and translated to an average of 27% fewer prey being consumed compared to independent predator effects. Non-independence of this magnitude is likely to have population level consequences for the prey and influence the distribution or prey preference of predators. Closer inspection shows that much of the risk reduction in this system is weak, to the point of being indistinguishable from independent predator effects, while few effects are strong. This pattern of many weak interactions and few strong ones parallels the pattern of interaction strengths documented previously in intertidal communities. Consequently, understanding strong interactors in multiple predator systems may help us understand the importance of a species.


Which are best for successful aging prediction? Bagging, boosting, or simple machine learning algorithms?
The worldwide society is currently facing an epidemiological shift due to the significant improvement in life expectancy and increase in the elderly population. This shift requires the public and scientific community to highlight successful aging (SA), as an indicator representing the quality of elderly people's health. SA is a subjective, complex, and multidimensional concept; thus, its meaning or measuring is a difficult task. This study seeks to identify the most affecting factors on SA and fed them as input variables for constructing predictive models using machine learning (ML) algorithms. Data from 1465 adults aged ≥ 60 years who were referred to health centers in Abadan city (Iran) between 2021 and 2022 were collected by interview. First, binary logistic regression (BLR) was used to identify the main factors influencing SA. Second, eight ML algorithms, including adaptive boosting (AdaBoost), bootstrap aggregating (Bagging), eXtreme Gradient Boosting (XG-Boost), random forest (RF), J-48, multilayered perceptron (MLP), Naïve Bayes (NB), and support vector machine (SVM), were trained to predict SA. Finally, their performance was evaluated using metrics derived from the confusion matrix to determine the best model. The experimental results showed that 44 factors had a meaningful relationship with SA as the output class. In total, the RF algorithm with sensitivity = 0.95 ± 0.01, specificity = 0.94 ± 0.01, accuracy = 0.94 ± 0.005, and F-score = 0.94 ± 0.003 yielded the best performance for predicting SA. Compared to other selected ML methods, the effectiveness of the RF as a bagging algorithm in predicting SA was significantly better. Our developed prediction models can provide, gerontologists, geriatric nursing, healthcare administrators, and policymakers with a reliable and responsive tool to improve elderly outcomes.


Molecular Survey and Microscopic Examination of Haemoparasites Infecting Lizards from Morocco.
Understanding the factors influencing parasite diversification, their distribution and infection patterns among hosts provides valuable information on the impact of parasitism on natural populations and the dynamics and evolution of disease outbreaks. In this study, we investigated prevalence and intensity of infection by blood parasites, the diversity, distribution, host specificity and infection patterns of apicomplexan parasites in lizards of Morocco. Samples from lizard specimens were collected between 2019 and 2021 from different prospected localities in Morocco. In total, 1500 lizards were collected, and their blood and tail tips were screened for blood parasites. Using a PCR approach, we detected two distinct apicomplexan parasite groups (haemogregarines and haemococcidians) and also Stramenopiles (Proteromonas). A significant difference was identified between hosts families, with no agamids or skinks infected and with particularly high infection rates in lacertids. Twenty-four samples were sequenced and included in a phylogenetic assessment. Two samples of Atlantolacerta andreanskyi were infected with unidentified species of Proteromonas. Three samples were infected with haemococcidians, including two samples with Schellackia and another sample with Lankesterella, more closely related to others isolated from bird hosts. These are the first reports of Lankesterella and Proteromonas in these lizards from Morocco. Nineteen different isolates of the complex of Hepatozoon/Karyolysus were identical or very similar to previously sequenced parasites from North Africa and Iberian Peninsula and included new host species. Our results highlight the need for future research on the vectors involved in transmission that may influence the ecology and distribution patterns of these parasites. Given that Morocco is a hot spot of endemicity and reptile richness, our results are relevant not only in the context of their taxonomy, but can also have implications for conservation.


Three-dimensional correction of scoliosis by a double spring reduction system as a dynamic internal brace: a pre-clinical study in Göttingen minipigs.
Adolescent idiopathic scoliosis (AIS) is a major skeletal deformity that is characterized by a combination of apical rotation, lateral bending and apical lordosis. To provide full 3D correction, all these deformations should be addressed. We developed the Double Spring Reduction (DSR) system, a (growth-friendly) concept that continuously corrects the deformity through two different elements: A posterior convex Torsional Spring Implant (TSI) that provides a derotational torque at the apex, and a concave Spring Distraction System (SDS), which provides posterior, concave distraction to restore thoracic kyphosis. To determine whether the DSR components are able to correct an induced idiopathic-like scoliosis and to compare correction realized by the TSI alone to correction enforced by the complete DSR implant. Preclinical randomized animal cohort study. Twelve growing Göttingen minipigs. Coronal Cobb angle, T10-L3 lordosis/kyphosis, apical axial rotation, relative anterior lengthening. All mini-pigs received the TSI with a contralateral tether to induce an idiopathic-like scoliosis with apical rotation (mean Cobb: 20.4°; mean axial apical rotation: 13.1°, mean lordosis: 4.9°). After induction, the animals were divided into two groups: One group (N=6) was corrected by TSI only (TSI only-group), another group (N=6) was corrected by a combination of TSI and SDS (DSR-group). 3D spinal morphology on CT was compared between groups over time. After 2 months of correction, animals were euthanized. Both intervention groups showed excellent apical derotation (TSI only-group: 15.0° to 5.4°; DSR-group: 11.2° to 3.5°). The TSI only-group showed coronal Cobb improvement from 22.5° to 6.0°, while the DSR-group overcorrected the 18.3° Cobb to -9.2°. Lordosis was converted to kyphosis in both groups (TSI only-group: -4.6° to 4.3°; DSR-group: -5.2° to 25.0°) which was significantly larger in the DSR-group (p<.001). The TSI alone realized strong apical derotation and moderate correction in the coronal and sagittal plane. The addition of distraction on the posterior concavity resulted in more coronal correction and reversal of induced lordosis into physiological kyphosis. This study shows that dynamic spring forces could be a viable method to guide the spine towards healthy alignment, without fusing it or inhibiting its growth.


Microglial migration mediated by ATP-induced ATP release from lysosomes.
Microglia are highly motile cells that act as the main form of active immune defense in the central nervous system. Attracted by factors released from damaged cells, microglia are recruited towards the damaged or infected site, where they are involved in degenerative and regenerative responses and phagocytotic clearance of cell debris. ATP release from damaged neural tissues has been suggested to mediate the rapid extension of microglial process towards the site of injury. However, the mechanisms of the long-range migration of microglia remain to be clarified. Here, we found that lysosomes in microglia contain abundant ATP and exhibit Ca(2+)-dependent exocytosis in response to various stimuli. By establishing an efficient in vitro chemotaxis assay, we demonstrated that endogenously-released ATP from microglia triggered by local microinjection of ATPγS is critical for the long-range chemotaxis of microglia, a response that was significantly inhibited in microglia treated with an agent inducing lysosome osmodialysis or in cells derived from mice deficient in Rab 27a (ashen mice), a small GTPase required for the trafficking and exocytosis of secretory lysosomes. These results suggest that microglia respond to extracellular ATP by releasing ATP themselves through lysosomal exocytosis, thereby providing a positive feedback mechanism to generate a long-range extracellular signal for attracting distant microglia to migrate towards and accumulate at the site of injury.


Patient Perceptions of Treatment Delivery Platforms for Cognitive Behavioral Therapy for Insomnia.
Stepped care has given rise to the proliferation of abbreviated CBT-I programs and delivery formats. This includes interventions delivered by allied health professionals and those delivered electronically through the Internet. This article aims to explore patient perceptions between electronic and face-to-face (FTF) delivery platforms for (abbreviated) CBT-I. Patients with insomnia from specialist sleep or psychology clinics and those from the general community in Sydney, Australia. Semistructured interviews were conducted with patients with insomnia, guided by a schedule of questions and a choice task to explore patient perceptions of the different CBT-I treatment delivery platforms (e.g., perceived advantages and disadvantages or willingness to engage with either platform). Interviews were transcribed verbatim and analyzed using Framework Analysis. Participants also completed a battery of clinical mood and insomnia measures. Fifty-one interviews were conducted with patients with insomnia from specialist sleep or psychology clinics (n = 22) and the general community (n = 29). Synthesis of the qualitative data set revealed three themes pertinent to the patients' perspective toward electronic and FTF CBT-I delivery: Concepts of Efficacy, Concerns About Treatment, and Treatment on My Terms. Participants' choice to engage with either platform was also informed by diverse factors including perceived efficacy of treatment, personal commitments, lifestyle, and beliefs about sleep and insomnia. Clarifying patient treatment priorities and allaying potential concerns about engaging with an electronic treatment platform represent important steps for disseminating eCBT-I into mainstream practice.


[Study on digitization of difference in drug color and odor of Magnoliae Officinalis Cortex before and after perspiration].
To digitalize the changes in characters of Magnoliae Officinalis Cortex after perspiration with colorimeter and electronic nose. With perspired and non-perspired Magnoliae Officinalis Cortex as objective, colorimeter and electronic nose were used to detect their color characteristic parameter and odor characteristic parameter. Finally, an identification model was established. In terms of drug color, the color characteristic parameter model was established for perspired and non-perspired Magnoliae Officinalis Cortex on the basis of L*, a*, b* color spaces. The range of 90% of reference values of perspired Magnoliae Officinalis Cortex: L* (52.22-59.42), a* (5.36-7.68), b* (22.04-27.05). The range of 90% of reference values of non-perspired Magnoliae Officinalis Cortex: L* (38.42-47.31), a* (9.63-11.85), b* (18.48-25.53). In terms of drug odor, the principal component analysis (PCA) and the partial least squares method (PLS) showed significant difference between perspired and non-perspired Magnoliae Officinalis Cortex. The difference in drug color and odor of Magnoliae Officinalis Cortex before and after perspiration can be digitalized according to color and odor characteristic parameters tested with colorimeter and electronic nose.


Protocol of an expertise based randomized trial comparing surgical Venae Sectio versus radiological puncture of Vena Subclavia for insertion of Totally Implantable Access Port in oncological patients.
Totally Implantable Access Ports (TIAP) are being extensively used world-wide and can be expected to gain further importance with the introduction of new neoadjuvant and adjuvant treatments in oncology. Two different techniques for the implantation can be selected: A direct puncture of a central vein and the utilization of a Seldinger device or the surgical Venae Sectio. It is still unclear which technique has the optimal benefit/risk ratio for the patient. A single-center, expertise based randomized, controlled superiority trial to compare two different TIAP implantation techniques. 100 patients will be included and randomized pre-operatively. All patients aged 18 years or older scheduled for primary elective implantation of a TIAP under local anesthesia who signed the informed consent will be included. The primary endpoint is the primary success rate of the randomized technique. Control Intervention: Venae Sectio will be employed to insert a TIAP by a surgeon; Punction of V. Subclavia will be used to place a TIAP by a radiologist. Duration of study: Approximately 10 months, follow up time: 90 days. The PORTAS 2 - Trial will be conducted in accordance with the protocol and in compliance with the moral, ethical, and scientific principles governing clinical research as set out in the Declaration of Helsinki (1989) and Good Clinical Practice (GCP). The Center of Clinical Trials at the Department of Surgery, University Hospital Heidelberg is responsible for design and conduct of the trial including randomization and documentation of patients' data. Data management and statistical analysis will be performed by the independent Institute for Medical Biometry and Informatics (IMBI), University of Heidelberg. The trial is registered at ClinicalTrials.gov (NCT00600444).


A GT dinucleotide repeat polymorphism in intron 1 of the H-cadherin (CDH13) gene.
A polymorphic dinucleotide (GT) repeat sequence was isolated from a genomic clone containing the human H-cadherin (CDH13) gene at 16q24.1-q24.2. This polymorphic marker will be a useful tool in the genetic study of various cancers.


Expression, purification, and biochemical properties of arginase from Bacillus subtilis 168.
The arginine-degrading and ornithine-producing enzymes arginase has been used to treat arginine-dependent cancers. This study was carried out to obtain the microbial arginase from Bacillus subtilis, one of major microorganisms found in fermented foods such as Cheonggukjang. The gene encoding arginase was isolated from B. subtilis 168 and cloned into E. coli expression plasmid pET32a. The enzyme activity was detected in the supernatant of the transformed and IPTG induced cell-extract. Arginase was purified for homogeneity from the supernatant by affinity chromatography. The specific activity of the purified arginase was 150 U/mg protein. SDS-PAGE analysis revealed the molecular size to be 49 kDa (Trix·Tag, 6×His·Tag added size). The optimum pH and temperature of the purified enzyme with arginine as the substrate were pH 8.4 and 45°C, respectively. The Km and Vmax values of arginine for the enzyme were 4.6 mM and 133.0 mM/min/mg protein respectively. These findings can contribute in the development of functional fermented foods such as Cheonggukjang with an enhanced level of ornithine and pharmaceutical products by providing the key enzyme in arginine-degradation and ornithine-production.


[Estimation of glycated hemoglobin values as a mathematical model].
By using various assays of glycated hemoglobin, the author deduced a model of this index by its level, association with major metabolic parameters from the linear and nonlinear correlations and regression. The duration of diabetes mellitus, serum total cholesterol, and postprandial glycemia were found to have the most considerable impact on the formation of different models of this index.


A possible association between exposure to n-hexane and parkinsonism.
Recently, some case-control studies and case reports have shown an association between solvent exposure and parkinsonisms. We present a 55-year-old male parkinsonian patient with chronic exposure to n-hexane for 17 years. The results of neurophysiological (electromyography, evoked potentials), neuroradiological (MRI) and neuropsychological tests performed on the patient suggest a role of this solvent at the level of the central nervous system. Biological susceptibility to neurotoxic compounds is discussed briefly.


Loading enhances the occurrence of startle responses in leg muscles.
The startle reflex is an involuntary reaction to sudden sensory input and consists of a generalized flexion response. Startle responses in distal leg muscles occur more frequently during standing compared to sitting. We hypothesized that sensory input from load receptors modulates the occurrence of startle responses in leg muscles. We administered sudden startling auditory stimuli (SAS) to 11 healthy subjects while (1) sitting relaxed, (2) standing relaxed, (3) standing while bearing 60% of their weight on the right leg, (4) standing while bearing 60% of their weight on the left leg, and (5) standing with 30% body weight support ('bilateral unloaded'). The requested weight distribution for each condition was verified using force plates. Electromyography data were collected from both tibialis anterior (TA) and the left sternocleidomastoid muscles. In the TA, startle responses occurred much more frequently during normal standing (26% of trials) compared to both sitting (6% of trials, p<0.01) and bilateral unloading (3% of trials, p<0.01). In the asymmetrical stance conditions, startle responses in the TA were more common in the loaded leg (21% of trials) compared to the unloaded leg (10% of trials, p<0.05). The occurrence of startle responses in the leg muscles was strongly influenced by load. Hence, it is likely that information from load receptors influences startle response activity. We suggest that, in a stationary position, startling stimuli result in a descending volley from brainstem circuits, which is gated at the spinal level by afferent input from load receptors.


Low doses of paraquat and polyphenols prolong life span and locomotor activity in knock-down parkin Drosophila melanogaster exposed to oxidative stress stimuli: implication in autosomal recessive juvenile parkinsonism.
Previous studies have shown that polyphenols might be potent neuroprotective agents in Drosophila melanogaster wild type Canton-S acutely or chronically treated with paraquat (PQ), a selective toxin for elimination of dopaminergic (DAergic) neurons by oxidative stress (OS), as model of Parkinson's disease (PD). This study reports for the first time that knock-down (K-D) parkin Drosophila melanogaster (TH-GAL4; UAS-RNAi-parkin) chronically exposed to PQ (0.1-0.25 mM), FeSO(4) (Fe, 0.1mM), deferoxamine (DFO, 0.01 mM) alone or (0.1mM) PQ in combination with polyphenols propyl gallate (PG, 0.1mM) and epigallocathecin gallate (EGCG, 0.1, 0.5mM) showed significantly higher life span and locomotor activity than untreated K-D flies or treated with (1, 5, 20mM) PQ alone. Whilst gallic acid (GA, 0.1, 0.5mM) alone or in the presence of PQ provoked no effect on K-D flies, epicathecin (EC, 0.5mM) only showed a positive effect on prolonging K-D flies' life span. It is shown that PG (and EGCG) protected protocerebral posterolateral 1 (PPL1) DAergic neurons against PQ. Interestingly, the protective effect of low PQ concentrations, DFO and iron might be explained by a phenomenon known as "hormesis." However, pre-fed K-D flies with (0.1mM) PQ for 7 days and then exposed to (0.25 mM) for additional 8 days affect neither survival nor climbing of K-D Drosophila compared to flies treated with (0.25 mM) PQ alone. Remarkably, K-D flies treated with 0.1mM PQ (7 days) and then with (0.25 mM) PQ plus PG (8 days) behaved almost as flies treated with (0.25 mM) PQ. Taken these data suggest that antioxidant supplements that synergistically act with low pro-oxidant stimuli to prolong and increase locomotor activity become inefficient once a threshold of OS has been reached in K-D flies. Our present findings support the notion that genetically altered Drosophila melanogaster as suitable model to study genetic and environmental factors as causal and/or modulators in the development of autosomal recessive juvenile Parkinsonism (AR-JD)/PD. Most importantly, we have shown for the first time that low amounts of stressors induce a health-promoting extending effect in K-D parkin flies. Altogether our present results open new avenues for the screening, testing and development of novel antioxidant drugs against OS stimuli in neurodegenerative disorders.


Ionic-size dependent electroosmotic flow in ion-selective biomimetic nanochannels.
Electrokinetic phenomena, especially electroosmosis in ion-selective environments, play a key role in many systems, from ion-selective nanopores to cellular processes. In this paper, the impact of ionic size on the electroosmotic flow through an ion-selective soft slit nanochannel is analytically studied. Meanwhile, the modified Poisson-Boltzmann and the modified Navier-Stokes equations were used for modeling the electrostatics and the electrohydrodynamics of the problem, respectively, and the derived equations were solved by linearizing method. The results reveal the importance of considering the effect of ionic size in the calculation, as the steric effects, especially at high charge densities of polyelectrolytes (PELs), dramatically alter both the ions arrangement and the electric potential; and amplify the electroosmotic flow. Considering Debye-Huckel parameters of 4 and 10 for the electrolyte layer and the PEL, respectively, we demonstrate that the dimensionless electroosmotic velocity in a soft nanochannel having a dimensionless soft layer thickness of 0.2, from 3.2 by ignoring the steric effect, can reach the value of 6 by considering the steric effect of ν=0.3.


Modulation of the catalytic properties of alpha-chymotrypsin by chemical modification at Tyr 146.
alpha--Chymotrypsin was modified with three different diazonium salts derived from D--phenylalanine. All three reagents react selectively with tyrosine 146 on the surface of the enzyme. The spectral and enzymatic properties of the azochymotrypsins are characteristic for each of the proteins. Dependent on the structure of the reagent employed for modification, the activity towards rho-nitroanilide substrate can be higher or about the same as that of the native enzyme.


Rescue of vaccinia virus lacking the E3L gene by mutants of E3L.
Vaccinia virus with the E3L gene deleted was able to replicate in RK-13 but not HeLa cells. This host range phenotype could be complemented by an E3L gene expressed transiently from a plasmid. Analysis of mutants of E3L indicates that the ability to complement deletion of E3L correlates with the ability of mutated proteins to bind double-stranded RNA but not with their ability to migrate to the nucleus.


Enhanced simultaneous absorption of NOx and SO2 in oxidation-reduction-absorption process with a compounded system based on Na2SO3.
Oxidation of sulfite and competitive absorption existed in Na2SO3 solution for simultaneous removal of NOx and SO2, inhibited the long-term high-efficiency when used for practical applications. A matching strategy was developed to solve these problems. Antioxidants combination was used to retard the oxidation of antioxidant and enhance inhibition of S(IV) (tetravalent sulfur) oxidation. Hydroquinone (HQ) and sodium thiosulfate (ST) showed a positive synergistic effect on inhibition of S(IV) oxidation. When SO2 concentration was 500 and 2000 ppmV, the addition of 0.1 wt.% HQ and 1 wt.% ST decreased the percentage of S(IV) oxidized by oxygen by over 30% and 40%, respectively. Alkali (Na2CO3) alleviated the competitive absorption between NOx and SO2. Moreover, Na2CO3 exhibited an enhancement effect on the absorption of NOx and SO2 when coupled with anti-oxidants. While the increase of oxygen pressure accelerated the oxidation of S(IV), the anti-oxidants can retard the oxidation. The measurement of pH suggested the removal efficiency of NOx highly depended on SO32⁻ concentration rather than pH. The further investigation of the mechanism suggested the match effect was related to the interaction between ST and the intermediate product of HQ. The match strategy holds a potential for application of SO32⁻ to denitration.


Prevalence of circumcision among men and boys aged 14 to 59 years in the United States, National Health and Nutrition Examination Surveys 2005-2010.
In 2009, an estimated 3590 new heterosexually acquired HIV infections occurred in males in the United States. Three randomized controlled trials demonstrated that male circumcision decreased a man's risk for HIV acquisition through heterosexual sex. We describe circumcision prevalence in US males and determine circumcision prevalence among males potentially at increased risk for heterosexually acquired HIV infection. We estimated circumcision prevalence among men and boys aged 14 to 59 years using data from the National Health and Nutrition Examination Surveys 2005-2010. We defined men and boys with 2 or more female partners in the last year as potentially at increased risk for heterosexually acquired HIV infection. Estimated circumcision prevalence was 80.5%. Prevalence varied significantly by year of birth, race/ethnicity, health insurance type, and family income. Circumcision prevalence among men and boys reporting 2 or more female partners in the last year was 80.4%, which corresponded to an estimated 3.5 million uncircumcised men and boys potentially at increased risk for heterosexually acquired HIV infection. Of these men and boys, 48.3% lacked health insurance. Circumcision prevalence in the United States differs by demographic group, and half of uncircumcised men and boys potentially at increased risk for heterosexually acquired HIV are uninsured. These data could inform recommendations and cost analyses concerning circumcision in the United States.


Determination of N,N-dimethyltryptamine in beverages consumed in religious practices by headspace solid-phase microextraction followed by gas chromatography ion trap mass spectrometry.
A novel analytical approach combining solid-phase microextraction (SPME)/gas chromatography ion trap mass spectrometry (GC-IT-MS) was developed for the detection and quantification N,N-dimethyltryptamine (DMT), a powerful psychoactive indole alkaloid present in a variety of South American indigenous beverages, such as ayahuasca and vinho da jurema. These particular plant products, often used within a religious context, are increasingly consumed throughout the world following an expansion of religious groups and the availability of plant material over the Internet and high street shops. The method described in the present study included the use of SPME in headspace mode combined GC-IT-MS and included the optimization of the SPME procedure using multivariate techniques. The method was performed with a polydimethylsiloxane/divinylbenzene (PDMS/DVB) fiber in headspace mode (70 min at 60 °C) which resulted in good precision (RSD<8.6%) and accuracy values (71-109%). Detection and quantification limits obtained for DMT were 0.78 and 9.5 mg L(-1), respectively and good linearity (1.56-300 mg L(-1), r(2)=0.9975) was also observed. In addition, the proposed method showed good robustness and allowed for the minimization of sample manipulation. Five jurema beverage samples were prepared in the laboratory in order to study the impact of temperature, pH and ethanol on the ability to extract DMT into solution. The developed method was then applied to the analysis of twelve real ayahuasca and vinho da jurema samples, obtained from Brazilian religious groups, which revealed DMT concentration levels between 0.10 and 1.81 g L(-1).


Intramyocardial left ventricle-to-coronary artery stent: a novel approach for the treatment of coronary artery disease.
The direct intramyocardial left ventricle-to-coronary artery stent may provide an optional minimally invasive technique for coronary artery bypass graft surgery. We seek to test whether blood flow and regional myocardial function improve with this stent in totally ischemic myocardium. The stent device was implanted in 8 anesthetized dogs using an open chest approach, arteriotomy of the proximal left anterior descending coronary artery, and connection of the vessel to the left ventricular chamber. Regional coronary blood flow and myocardial function were monitored under three conditions: normal coronary flow (baseline), coronary ligation, and stent flow. Left anterior descending coronary ligation markedly reduced coronary artery blood flow and regional myocardial function. With flow solely from the stent, the blood flow pattern changed to one with high peak forward flow during systole compared with baseline (94.8 +/- 48.9 versus 56.8 +/- 21.1 mL/min; p < 0.05) and one with significant negative backflow during diastole compared with baseline (-37.4 +/- 23.1 versus 11.3 +/- 17.2 mL/min; p < 0.05). However, the resultant mean forward flow increased to approximately 50% of baseline compared with less than 5% of baseline after coronary ligation. Regional myocardial function diminished entirely after coronary ligation, but recovered to approximately 60% of baseline with the stent. Normal systemic hemodynamics and global ventricular contractile function were maintained with the stent. The left ventricle-to-coronary artery stent is a simple and readily deployable device that allows the perfusion of epicardial vessels directly from the left ventricle and can provide significant blood flow to improve the performance of ischemic myocardium. It may provide an effective, alternative means of treating coronary artery disease when standard coronary artery bypass graft surgery is not suitable.


Diversity of Babesia spp. in cervid ungulates based on the 18S rDNA and cytochrome c oxidase subunit I phylogenies.
Free ranging ungulates, represented in Europe mostly by several deer species, are important hosts for ticks and reservoirs of tick-borne infections. A number of studies have focused on the prevalence of tick borne pathogens in deer chiefly with the aim to determine their potential role as reservoir hosts for important human and livestock pathogens. However, genetic similarity of Babesia spp. forming a group commonly termed as a clade VI that accommodates the deer piroplasms, complicates this task and has led to the description of a bewildering array of poorly characterised strains. This study aims to resolve this issue by using two independent genetic loci, nuclear 18S rRNA and mitochondrial cytochrome c oxidase subunit I genes, used in parallel to identify Babesia isolates in free-ranging red, sika, and roe deer in two areas of their co-occurrence in the Czech Republic. The COX1 loci, in contrast to 18S rRNA gene, shows a clear difference between interspecific and intraspecific variation at the nucleotide level. The findings confirm B. divergens, Babesia sp. EU1 and B. capreoli in studied deer species as well as common presence of another unnamed species that matches a taxon previously referred to as Babesia sp. or Babesia cf. odocoilei or Babesia CH1 group in several other sites throughout Europe. The invasive sika deers enter the life cycle of at least three piroplasmid species detected in native deer fauna. The presence of B. divergens in both sika and red deer in an area where bovine babesiosis is apparently absent raises important questions regarding the epidemiology, host specificity and taxonomic status of the parasite.


Synchronizing Mammalian Cells for Mitotic Analysis of the Localization of Survivin.
This paper describes an easy method to enrich the harvest of adherent mammalian cells at each stage of mitosis (from prometaphase to cytokinesis) by combining Eg5 inhibition using dimethylenastron (DMA) with mitotic shake-off, followed by timed release from the drug.


Where should patients with or at risk of delirium be treated in an acute care system? Comparing the rates of delirium in patients receiving usual care vs alternative care: A systematic review and meta-analysis.
Delirium is an acute condition that occurs in hospitalised patients and leads to poor patient outcomes that can last long term. Therefore, the importance of prevention is undeniable and adopting new models of care for at-risk patients should be prioritised. This systematic review and meta-analysis will assess the effectiveness of different interventions designed to prevent or manage delirium in acutely unwell hospitalised patients. MEDLINE, EMBASE, PsycINFO, OpenGrey, Web of Science and reference lists of journals were searched. Eligible studies reported on incidence or duration of delirium, used a validated delirium diagnostic tool and compared an intervention to either a control or another intervention group. Meta-analyses were conducted, and GRADEpro software was used to assess the certainty of evidence. This review is registered on PROSPERO. A total of 59 studies were included and 33 were eligible for meta-analysis. Delirium incidence was most significantly reduced by non-pharmacological multicomponent interventions compared with usual care, with pooled risk ratios of 0.57 (95% CI: 0.44 to 0.73, 10 randomised controlled trials) and 0.47 (95% CI: 0.35 to 0.64, six observational studies). Single-component interventions did not significantly reduce delirium incidence compared with usual care in seven randomised trials (risk ratio = 0.92, 95% CI: 0.81 to 1.04). The most effective single-component intervention in reducing delirium incidence was a hospital-at-home intervention (risk ratio = 0.29, 95% CI: 0.09 to 0.87). Non-pharmacological multicomponent interventions are effective in preventing delirium; however, the same cannot be said for other interventions because of uncertain results. There is some evidence that providing multicomponent interventions in patients' homes is more effective than in a hospital setting. Therefore, researching the benefits of hospital-at-home interventions in delirium prevention is recommended.


Biomethane is produced by acetate cleavage, not direct interspecies electron transfer: genome-centric view and carbon isotope.
Understanding the source of methane (CH4) is of great significance for improving the anaerobic fermentation efficiency in bioengineering, and for mitigating the emission potential of natural ecosystems. Microbes involved in the process named direct interspecies electron transfer coupling with CO2 reduction, i.e., electrons released from electroactive bacteria to reduce CO2 into CH4, have attracted considerable attention for wastewater treatment in the past decade. However, how the synergistic effect of microbiota contributes to this anaerobic carbon metabolism accompanied by CH4 production still remains poorly understood, especial for wastewater with antibiotic exposure. Results show that enhancing lower-abundant acetoclastic methanogens and acetogenic bacteria, rather than electroactive bacteria, contributed to CH4 production, based on a metagenome-assembled genomes network analysis. Natural and artificial isotope tracing of CH4 further confirmed that CH4 mainly originated from acetoclastic methanogenesis. These findings reveal the contribution of direct acetate cleavage (acetoclastic methanogenesis) and provide insightsfor further regulation of methanogenic strategies.


Global public health surveillance under new international health regulations.
The new International Health Regulations adopted by the World Health Assembly in May 2005 (IHR 2005) represents a major development in the use of international law for public health purposes. One of the most important aspects of IHR 2005 is the establishment of a global surveillance system for public health emergencies of international concern. This article assesses the surveillance system in IHR 2005 by applying well-established frameworks for evaluating public health surveillance. The assessment shows that IHR 2005 constitutes a major advance in global surveillance from what has prevailed in the past. Effectively implementing the IHR 2005 surveillance objectives requires surmounting technical, resource, governance, legal, and political obstacles. Although IHR 2005 contains some provisions that directly address these obstacles, active support by the World Health Organization and its member states is required to strengthen national and global surveillance capabilities.


Use of nesiritide (human B-type natriuretic peptide) in infants following cardiac surgery.
We report the use of nesiritide in two infants following cardiac surgery. Both infants had increased intracardiac filling pressures postoperatively, despite traditional afterload reduction and diuretics. Both infants demonstrated clinical improvement with nesiritide therapy. There is currently limited data available describing the use of nesiritide in infants.


A qualitative study of UK pharmacy pre-registration graduates' views and reflections on pharmacist prescribing.
Pre-registration pharmacy graduates are a key group in relation to developing and implementing innovations in practice. To explore the views and reflections on pharmacist prescribing of UK pre-registration pharmacy graduates. Semi-structured telephone interviews with a sample (n = 12) of pre-registration pharmacy graduates. Most reported a desire to train as prescribers, largely for reasons of professional development, but acknowledged the need first to develop as pharmacists. They perceived the lack of organisational strategy, pharmacist prescribers' self-confidence and additional workload to be barriers and were aware of the value of interprofessional relationships as potential facilitators to pharmacist prescribing. While participants were supportive of pharmacist prescribing, they highlighted the need first to gain experience as pharmacists and were acutely aware of barriers to prescribing implementation.


Promoter methylation regulates cadherin switching in squamous cell carcinoma.
Cadherins are cell adhesion molecules that modulate the epithelial phenotype and regulate tumor invasion. To identify the role of promoter methylation in regulating E-cadherin expression and in the "switching" of cadherins in oral squamous cell carcinoma (SCC), we studied 14 cell lines for cadherin expression. Immunoblotting revealed that only two (HOC-313 and HA-376) showed strong up-regulation of N-cadherin, and neither expressed E-cadherin. These results were confirmed by PCR. Furthermore, analysis of genomic DNA showed that the lack of E-cadherin expression in the two cell lines was not due to gene deletion. In both cell lines, methylation-specific PCR indicated extensive methylation of the 5' CpG island in the E-cadherin promoter. After treatment with a DNA methylation inhibitor (5-Aza-2-deoxycytidine), both immunoblotting and immunofluorescence staining showed that HA-376 cells newly expressed E-cadherin with a parallel decrease in their N-cadherin expression. Multiplex RT-PCR demonstrated that the down-regulation of N-cadherin mRNA was coordinately regulated with E-cadherin expression. Thus, methylation of the 5' CpG island in the E-cadherin promoter induces reciprocal expression of E- and N-cadherins in oral SCC by an unknown mechanism that appears to be mediated at the level of N-cadherin gene expression. These events may play an important role in the regulation of tumor cell mobility and invasion.


Identification of underexplored mesenchymal and vascular-related cell populations in human skeletal muscle.
Skeletal muscle repair and maintenance are directly and indirectly supported by interstitial cell populations such as vascular cells and fibro-adipogenic progenitors (FAPs), a subset of which express Twist2 and possess direct myogenic potential. Furthermore, work in rodents has highlighted the potential of pericytes to act as progenitor cells, giving rise to muscle cells and transdifferentiating into endothelial cells. However, less is understood about these populations in human skeletal muscle. Here, we performed single-cell RNA sequencing (scRNAseq) on ∼2,000 cells isolated from the human semitendinosus muscle of young individuals. This demonstrated the presence of a vascular-related cell type that expressed pericyte and pan-endothelial genes that we localized to large blood vessels within skeletal muscle cross sections and termed endothelial-like pericytes (ELPCs). RNA velocity analysis indicated that ELPCs may represent a "transition state" between endothelial cells and pericytes. Analysis of published scRNAseq data sets revealed evidence for ELPCs in trunk and heart musculature, which showed transcriptional similarity. In addition, we identified a subset of FAPs expressing TWIST2 mRNA and protein. Human TWIST2-expressing cells were anatomically and transcriptionally comparable to mouse Twist2 cells as they were restricted to the myofiber interstitium, expressed fibrogenic genes but lacked satellite cell markers, and colocalized with the FAPs marker PDGFRα in human muscle cross sections. Taken together, these results highlight the complexity of stromal cells residing in human skeletal muscle and support the utility of scRNAseq for discovery and characterization of poorly described cell populations.


Structural and functional analysis of somatic coding and UTR indels in breast and lung cancer genomes.
Insertions and deletions (Indels) represent one of the major variation types in the human genome and have been implicated in diseases including cancer. To study the features of somatic indels in different cancer genomes, we investigated the indels from two large samples of cancer types: invasive breast carcinoma (BRCA) and lung adenocarcinoma (LUAD). Besides mapping somatic indels in both coding and untranslated regions (UTRs) from the cancer whole exome sequences, we investigated the overlap between these indels and transcription factor binding sites (TFBSs), the key elements for regulation of gene expression that have been found in both coding and non-coding sequences. Compared to the germline indels in healthy genomes, somatic indels contain more coding indels with higher than expected frame-shift (FS) indels in cancer genomes. LUAD has a higher ratio of deletions and higher coding and FS indel rates than BRCA. More importantly, these somatic indels in cancer genomes tend to locate in sequences with important functions, which can affect the core secondary structures of proteins and have a bigger overlap with predicted TFBSs in coding regions than the germline indels. The somatic CDS indels are also enriched in highly conserved nucleotides when compared with germline CDS indels.


A phase 2 study of the bivalent VLP norovirus vaccine candidate in older adults; impact of MPL adjuvant or a second dose.
Acute norovirus gastroenteritis causes significant morbidity and in uncommon cases fatality in older adults. We investigated the safety and immunogenicity of bivalent virus-like particle (VLP) vaccine candidate formulations with and without monophosphoryl lipid A (adjuvant MPL) in this population. In this phase II, double-blind, controlled trial 294 healthy adults, ≥ 60 years of age, were randomized (1:1:1:1) to four groups to receive one or two intramuscular immunizations 28 days apart, with 26 18-49 year-old controls who received one MPL-free dose. One-dose groups received placebo on Day 1. Vaccine formulations contained 15 μg GI.1 and 50 μg GII.4c VLP antigens and 500 μg Al(OH)3, with or without 15 μg MPL. We measured histo-blood group antigen blocking (HBGA) antibodies and ELISA Ig at Days 1, 8, 29, 57, 211 and 393, and avidity indices and cell-mediated immunity (CMI). Solicited local and systemic adverse events (AE) were assessed for 7 days and unsolicited AEs for 28 days after each injection. After one dose HBGA antibodies to both VLP antigens increased with similar kinetics and magnitude in all groups; geometric mean titres (GMTs) persisted above baseline through Day 393. GMTs were similar across age strata (18-49, 60-74, 75-84 and ≥ 85 years of age) and unaffected by a second vaccination or MPL. Total Ig showed similar responses. No clinically relevant differences or changes in avidity or CMI were observed between formulations. Both formulations were well tolerated with no vaccine-related SAEs, the most frequent AEs being mild injection site pain and fatigue. Adults over 60 years of age displayed no safety concerns and had similar immune responses to the norovirus VLP vaccine candidate as younger adults, unaffected by increasing age, a second dose or inclusion of MPL. This data supports the further development of the MPL-free vaccine candidate for older adults.


Subclinical Angioid Streaks with Pseudodrusen: A New Phenotype of Age-Related Macular Degeneration.
To describe subclinical angioid streaks (AS) as a frequent, peculiar age-related macular degeneration (AMD) phenotype, comparing features of eyes with subclinical AS with those of eyes with AMD without AS. This was a retrospective, observational study. Among a patient cohort with AMD, we selected patients without known causes for AS whose eyes showed signs of angioid streaks (AS) on structural optical coherence tomography (OCT) but not on fundus examination. Selected OCT features of AS were Bruch's membrane (BM) breaks and large BM dehiscences. Among 543 eyes of 274 patients with AMD (mean ± standard deviation: 82 ± 7 years), 73 eyes of 46 patients (81 ± 7 years; p = 0.432) showed AS features on OCT (OCT AS) that were not visible on fundus examination. Estimated prevalence of subclinical age-related AS was 13.4% (95% confidence interval 10.3-16.3%) in this AMD population. Fifty-three eyes (73%) with AS features were affected by peripapillary atrophy, often with a "petaloid-like" pattern, similar to typical features of AS disease. Almost all cases (97%) presented reticular pseudodrusen (RPD), with (41%) or without (59%) drusen showing a significant difference in RPD prevalence in OCT AS eyes in comparison to AMD eyes without subclinical AS using generalized estimating equations (P < 0.001). Among the 73 subclinical AS cases, 71 were affected by late AMD (57 with macular neovascularization, 14 with geographic atrophy), showing a more advanced AMD stage in comparison with AMD eyes without subclinical AS (P < 0.001). The following OCT features were disclosed: BM breaks in 100% of cases and BM dehiscences in 37%. Subclinical AS in eyes with AMD is a peculiar phenotype of the disease, with features suggesting a primary involvement of Bruch's membrane and clinical similarities with mild, late-onset pseudoxanthoma elasticum.


Expression of hepatitis B virus antigens in attenuated Salmonellae for oral immunization.
The aim of our work is to identify hepatitis B virus antigens that can be stably expressed in attenuated Salmonellae and elicit protective immune responses as live oral route vaccines. As a first carrier system, we expressed T-cell and B-cell epitopes of hepatitis B virus as fusion proteins with the non-toxic subunit B (LT-B) in attenuated Salmonellae. These recombinant Salmonellae elicited anti-LT-B T- and B-cell immune responses and anti-HBV nucleocapsid antigen (HBcAg) T-cell responses when fed to mice. To combine the protective potential and the high immunogenicity of HBc with the induction of virus neutralizing antibodies to HBV surface antigen, we constructed vectors expressing hybrid HBc/pre-S particles in which the pre-S epitopes were surface-exposed. With one of these vectors, stable constitutive high level expression of hybrid HBc/pre-S2 particles was achieved in several attenuated Salmonella strains. When recombinant Salmonellae expressing such hybrid HBc/pre-S2 fusion proteins were fed to mice, the animals developed high titres of anti-HBcAg-specific serum IgG after a single or multiple oral immunizations, depending on the strain used as a carrier. In addition, lower titered antibodies against the pre-S2 antibody-binding sites were elicited. This is the first HBV antigen eliciting high-titered immune responses after a single oral immunization in recombinant Salmonellae. The immunogenicity of periplasmic LT-B and cytoplasmic HBc/pre-S2 shows that surface exposure of a foreign antigen is not a prerequisite for its immunogenicity in live attenuated Salmonellae.


Unblending Borderline Personality and Bipolar Disorders.
Borderline Personality (BPD) and Bipolar (BP) disorders stimulate an academic debate between their distinction and the inclusion of Borderline in the Bipolar spectrum. Opponents to this inclusion attribute the important differences and possible diagnostic incomprehension to overlapping symptoms. We tested 248 Borderline and 113 Bipolar patients, consecutively admitted to the Psychiatric Unit, through DSM-IV Axis I and II Disorders (SCID-I/II), Hamilton Depression Rating Scale (HAM-D), Hamilton Anxiety Rating Scale (HAM-A), Young Mania Rating Scale (YMRS) and Borderline Personality Disorder Severity Index-IV (BPDSI-IV). All the tests statistically discriminated the disorders (p < 0.0001). Overlapping symptoms resulted significantly different (impulsivity = 5.32 in BPD vs 1.55 in BP, p < 0.0001; emotional instability = 7.11 in BPD vs 0.55 in BP, p < 0.0001) and the range of their scores gives the opportunity for an even more precise discrimination. Distinctive traits (e.g. irritability or sexual arousal) are also discussed in order to try to qualify the core of these disorders to a higher degree. Comorbidity proves to be extremely small (3.6%). However, Borderline patients with manic features offer a privileged point of view for a deeper analysis. This allows for the possibility of a more precise examination of the nature and load of each symptom. Borderline Personality and Bipolar Disorders can be distinguished with high precision using common and time-sparing tests. The importance of discriminating these clinical features may benefit from this evidence.


Structural analysis of ING3 protein and histone H3 binding.
Proteins belonging to the ING family regulate the transcriptional state of chromatin by recruiting remodeling complexes to sites with histone H3 trimethylated at Lysine 4 (H3K4me3). This modification is recognized by the Plant HomeoDomain (PHD) present at the C-terminal region of the five ING proteins. ING3 facilitates acetylation of histones H2A and H4 by the NuA4-Tip60 MYST histone acetyl transferase complex, and it has been proposed to be an oncoprotein. The crystal structure of the N-terminal domain of ING3 shows that it forms homodimers with an antiparallel coiled-coil fold. The crystal structure of the PHD is similar to those of its four homologs. These structures explain the possible deleterious effects of ING3 mutations detected in tumors. The PHD binds histone H3K4me3 with low-micromolar, and binds the non-methylated histone with a 54-fold reduced affinity. Our structure explains the impact of site directed mutagenesis experiments on histone recognition. These structural features could not be confirmed for the full-length protein as solubility was insufficient for structural studies, but the structure of its folded domains suggest a conserved structural organization for the ING proteins as homodimers and bivalent readers of the histone H3K4me3 mark.


Uterine rupture and perinatal outcome.
In view of the increasing number of caesarean sections (CS), we wanted to investigate the clinical aspects of uterine rupture including perinatal outcome. A retrospective investigation of 24,181 deliveries at Stockholm South General Hospital between 1999 and 2004. Patient notes from cases with ICD-codes 0710 and 0711 were studied together with charts from previous deliveries and neonatal data from the Paediatric Department. Some 22 cases of uterine rupture were found, giving an incidence of 0.9 per 1,000 deliveries. In all cases, the diagnosis was confirmed at laparotomy. In 19/22 cases, the rupture occurred in patients with a previous uterine scar, 18 of whom were delivered at term and one at 16 gestational weeks. One case of intrauterine fetal death was noted. Of the remaining 20 newborns, 9 had a 5-min Apgar </=7, one died after 3 days, and one had neurological sequele at follow-up. The remaining 18 newborns were healthy at discharge and up to 1 year of age. A previous uterine scar was the single most obvious risk factor. A positive finding was the relatively low risk of long-term post-asphyctic sequele in the newborns.


Factors Influencing the Sugar-Sweetened Beverage Intake of Caregivers of Adolescents in Appalachia.
To identify factors that influence the sugar-sweetened beverage (SSB) intake of caregivers of middle school-aged adolescents. Cross-sectional. Southwestern Virginia, US, part of Central Appalachia. Caregivers (n = 362) of adolescents enrolled in the Kids SIPsmartER trial. Participants were mostly female (91%) and non-Hispanic White (96%), and 21% received Supplemental Nutrition Assistance Program (SNAP) benefits. Caregiver daily SSB intake and demographics, personal-level, interpersonal-level, and environmental-level determinants. Descriptive statistics, 1-way ANOVA, and stepwise regression. On average, caregivers consumed 25.7 (SD, 33.2) fluid ounces of SSB per day. In the final model, which included all variables, age (β = -0.41; P < 0.05), receiving SNAP benefits (β = 14.19; P ≤ 0.01), behavioral intentions (β = -5.48; P ≤ 0.001), affective attitudes (β = -2.15, P < 0.05), perceptions of whether their adolescent frequently consumes high amounts of SSB (β = 1.92; P ≤ 0.001), and home availability (β = 7.43; P ≤ 0.01) were significantly associated with SSB intake. Caregivers of Appalachian middle school students are high SSB consumers. Findings highlight the importance of implementing behavioral interventions for caregivers of adolescents that target multiple levels of influence, including demographic, personal-level, interpersonal-level, and environmental-level factors. Interventions may be particularly important for communities and groups with higher SSB intakes, such as those in Appalachia and who receive SNAP benefits.


Immunohistochemical demonstration of insulin-like growth factor I in inflammatory lesions in Wegener's granulomatosis and idiopathic midline destructive disease.
The immunoreactivity of the trophic peptide insulin-like growth factor I (IGF-I; somatomedin C) was mapped in nasal mucosa biopsies from three patients with Wegener's granulomatosis (WG) and one with idiopathic midline destructive disease (IMDD; idiopathic midline granuloma). Strongly increased IGF-I immuno-reactivity restricted to cells bordering and in vessel walls and in granulomas (WG) was demonstrated, while necrotic and noninflammatory areas were negative. Treatment with steroids and cyclophosphamide reduced the IGF-I immunoreactivity. The abnormally increased IGF-I immunoreactivities in WG and IMDD probably reflects the reactive growth processes in diseased tissue and is not thought to be the primary cause of either disease. IGF-I may be formed locally by cells in and close to the vascular walls in areas with active disease resulting in e.g. vascular growth, granuloma formation, and finally vessel obliteration and necrosis. IGF-I is likely to form, possibly in concert with other trophic factors, a link in the chain of events resulting in the tissue abnormalities in WG and IMDD.