Home smoking restrictions before, during and after pregnancy-a qualitative study in rural China.
Worldwide, many nonsmokers (often women and children) are exposed to second-hand smoke (SHS) in home settings, as men retain their traditional power and control within their family and women and children have limited agency to intervene. This study, set up to explore home smoking management in rural China, found that some women were able to positively intervene to restrict men's smoking at three key stages: prior to conception, during their pregnancy and at the early years of their children's lives. By utilizing dominant social, health and political narratives about the importance of raising a healthy child supported by the One-Child Policy in China, combined with the fear of health risks of SHS to young children, the women were able to use their elevated status as bearer and carers of the only children to subvert the pre-eminence of men in domestic environments, enabling them to positively influence home smoking. While this study highlights the possibility for future smoking cessation initiatives in China by incorporating family carers' elevated awareness of protection of children's health in key stages of childhood, there is also a need for further health education, as family members were unsure why they needed to keep children smoke-free, which may partially explain why few households were smoke-free.


Effects of ethanol on early potassium currents in Aplysia: cell specificity and influence of channel state.
The effects of ethanol (EtOH) on the early potassium current, IA, were examined in 3 identified neurons of Aplysia using voltage-clamp techniques. The primary effect of EtOH on this current was a pronounced increase in the time constant of decay. However, this effect was cell specific, being evident in cells MCC and R15 but not in cell B1. Other parameters of IA were not greatly affected in any of the cells, in comparison with the effects on decay time constant. Baseline parameters of IA were measured in each of the cells to determine whether subpopulations of IA channel might exist, and be differentially sensitive to EtOH. While differences did appear among cells, they were not consistent with an explanation of EtOH's actions based upon distribution of channel subtypes. The effect of EtOH on IA decay was dependent upon the voltage-clamp protocol used. When inactivation of IA developed at -20 mV, the slower development of inactivation noted above occurred. When inactivation without channel opening was produced by means of a prepulse to -40 mV, EtOH speeded up the development of inactivation. A number of possible explanations for these findings are discussed. Most of the effects of EtOH occurred within 1 min of application of the drug, suggesting relatively rapid access to the site of action. Effects continued to develop over succeeding minutes. This slower-developing effect may reflect either a delayed access to channels due to slower diffusion into or lateral movement within the lipid phase of the membrane, or it may indicate that channels are accessible to the EtOH molecule only when in certain states.


Supplementing Closed Ecological Life Support Systems with In-Situ Resources on the Moon.
In this review, I explore a broad-based view of technologies for supporting human activities on the Moon and, where appropriate, Mars. Primarily, I assess the state of life support systems technology beginning with physicochemical processes, waste processing, bioregenerative methods, food production systems and the robotics and advanced biological technologies that support the latter. We observe that the Moon possesses in-situ resources but that these resources are of limited value in closed ecological life support systems (CELSS)-indeed, CELSS technology is most mature in recycling water and oxygen, the two resources that are abundant on the Moon. This places a premium on developing CELSS that recycle other elements that are rarified on the Moon including C and N in particular but also other elements such as P, S and K which might be challenging to extract from local resources. Although we focus on closed loop ecological life support systems, we also consider related technologies that involve the application of biological organisms to bioregenerative medical technologies and bioregenerative approaches to industrial activity on the Moon as potential future developments.


As-needed opioid dosing for emergency patients with a daily use of opioids.
Patients with a daily use of opioids have a higher risk of insufficient pain treatment during hospitalization than other patients. This study aimed to examine whether as-needed opioid doses (PRN) were adequately adjusted when patients were admitted to the emergency department (ED) with pain. Patients, with a daily use of opioids, who received PRN opioid within the first 3 h after admission at the ED at Aarhus University Hospital, Denmark, were prospectively included from February 2021 to June 2021. The primary outcome was the proportion of patients receiving an inadequate initial dose of PRN opioid, here defined as <15% of daily dose of opioids (DDO) based on sparse evidence, but aligning with the prevailing clinical practice. Secondary outcomes included risk of an inadequate PRN dose in relation to DDO (patients were dichotomized into two groups (DDO <60 or ≥60 mg/day). Among 252 patients admitted to the ED with a daily use of opioids, 149 were admitted with pain and 82 received a PRN opioid dose within 3 h. Twenty-seven out of 82 (33%) patients received a PRN dose of <15% of DDO (95% CI: 23.7-43). When dichotomised; 10 out of 50 (20%) patients with a DDO <60 mg/day (95% CI: 10.0-33.7) versus 17 out of 32 (53.1%) patients with a DDO ≥60 mg/day (95% CI: 34.7-70.9) received an inadequate PRN dose (relative risk, RR: 2.65 [95% CI: 1.4-5.1]). Patients with daily use of opioids presenting in the ED with acute pain had a high risk of inadequate PRN opioid dose, especially if the DDO was high. Awareness about and education focusing on sufficient PRN doses for patients with a daily use of opioids is (still) called for.


Serum uric acid: a futile bystander in endothelial function?
This study aims to investigate the relationship between serum uric acid levels and endothelial function, oxidative stress, and hemodynamic parameters, and to determine if uric acid levels provide additional insights beyond traditional factors like ageing and hypertension in volunteers with low cardiovascular risk factors. Serum uric acid is known for its antioxidant properties, but it may also contribute to cardiovascular risk. The study enrolled 40 male participants, divided into three groups based on age and blood pressure status. Group 1 comprised younger participants, group 2 included older individuals without hypertension, and group 3 consisted of older patients with hypertension. The study assessed endothelial function using laser Doppler imaging and measured acetylcholine- and sodium nitroprusside-induced hyperaemia. The heat microcirculatory response was also examined in the presence of L-NAME, an inhibitor of NOS synthase. The study evaluated oxidative stress and arterial stiffness by measuring allantoin, angiotensin II, Homocitrulline/Lysine, and Chloro-Tyrosine/Tyrosine ratios, as well as by performing non-invasive measurements of aortic augmentation indexes and carotid-femoral pulse wave velocity. The study found that uric acid levels did not differ significantly among the three groups. Augmentation indexes increased with ageing, but hypertension did not have an additional effect. Blood pressure and carotid-femoral pulse wave velocity differed among the groups, with the lowest values among younger participants and the highest values among older individuals with hypertension. Allantoin and angiotensin II levels did not differ among the groups. However, Homocitrulline/Lysine and Chloro-Tyrosine/Tyrosine ratios were significantly lower in young subjects. Correlation and multivariable analysis showed that uric acid had no effect on any of the studied parameters. Despite a strong association between ageing and systolic blood pressure with impaired endothelial function, oxidative stress, and arterial stiffness, only ageing retained a significant effect in the multivariable analysis. In healthy or hypertensive adults with normal renal function, serum uric acid appears to be a futile bystander in endothelial function, oxidative stress, and arterial stiffness, in contrast to ageing, which reduces NO bioavailability. This study suggests that traditional factors such as ageing and hypertension should be the focus of clinical assessment and management of cardiovascular risk, rather than uric acid levels.


Single turnovers of the EcoRI restriction endonuclease.
Single turnovers of the EcoRI restriction endonuclease, cleaving its recognition site on the covalently closed form of plasmid pMB9, were examined. Two methods were used to monitor the progress of the reactions: one involved quenching the reaction at various times followed by the electrophoretic separation of the products cleaved in one and in both strands of the duplex; the other employed a stopped-flow fluorimeter to measure the amount of ethidium bromide bound to the DNA as it changes when the DNA, cleaved in at least one strand, dissociates from the enzyme. Two procedures were used to initiate the reactions. For some, one solution containing the enzyme was mixed with a second containing both DNA and MgCl2: in these reactions, the fluorescence changed at the same rate as the cleavage of the first strand of the duplex. Other reactions were started by the addition of MgCl2 to a pre-equilibrium of enzyme and DNA: here, both strands of the DNA were cleaved faster than before, with the fluorescence signal now occurring at the same time as the cleavage of the second strand. The different kinetics from the two assays and the two mixing procedures are consistent with the rates of these reactions being controlled by protein conformational changes. These may affect either one subunit alone within the dimeric EcoRI enzyme, allowing the enzyme to cleave only one strand of the DNA in each turnover. Alternatively, both subunits of the dimer may change, so that the enzyme then cleaves both strands during the life-time of one enzyme-DNA complex.


Italian Physicians' Opinions on Rotavirus Vaccine Implementation.
Rotavirus (RV) infection is the main cause of severe acute gastroenteritis (GE) in the pediatric population and has a major impact in both developing and industrialized countries. The reduction of severe RVGE cases, followed by death or hospitalization, is considered the main benefit of RV vaccination, even though its implementation often faces obstacles. In Italy, the recently approved National Immunization Plan aims to overcome the differences among regions, offering a universal free RV vaccination. The aim of the study was to evaluate the opinions on benefit and acceptability of RV vaccination related to the perception of the burden of RV disease. Data were collected from 108 physicians in 2015 by a questionnaire consisting of 12 questions; some answers were compared with those obtained with a similar tool in 2011. The majority of respondents (76.2%) was convinced of the benefit of the vaccine and 57.4% recommended it routinely, but more than half indicated a <25% adherence to RV vaccination among their patients. As the main reasons of vaccine refusal, skepticism about the vaccine (60.4%) and its cost (34.1%) were indicated. Our data confirm that more information and counselling are needed to increase RV vaccine coverage.


Identifying priority medicines policy issues for New Zealand: a general inductive study.
To identify priority medicines policy issues for New Zealand. Stakeholders from a broad range of healthcare and policy institutions including primary, secondary and tertiary care. Exploratory, semistructured interviews were conducted with 20 stakeholders throughout New Zealand. The interviews were digitally recorded, transcribed and coded into INVIVO 10, then compared and grouped for similarity of theme. Perceptions, experiences and opinions regarding New Zealand's medicines policy issues were recorded. A large proportion of stakeholders appeared to be unaware of New Zealand's (NZ) medicines policy. In general, the policy was considered to offer consistency to guide decision-making. In the context of Pharmaceutical Management Agency's (PHARMAC's) fixed budget for procuring and subsidising medicines, there was reasonable satisfaction with the range of medicines available-rare disorder medicines being the clear exception. Concerns raised were by whom and how decisions are made and whether desired health outcomes are being measured. Other concerns included inconsistencies in evidence and across health technologies. Despite attempts to improve the situation, lower socioeconomic groups (including rural residents) Māori and Pacific ethnicities and people with rare disorders face challenges with regards to accessing medicines. Other barriers include, convenience to and affordability of prescribers and the increase of prescription fees from NZ$3 to NZ$5. Concerns related to the PHARMAC of New Zealand included: a constraining budget; non-transparency of in-house analysis; lack of consistency in recommendations between the Pharmacology and Therapeutics Advisory Committee. Constraints and inefficiencies also exist in the submission process to access high-cost medicines. The results suggest reasonable satisfaction with the availability of subsidised medicines. However, some of the major challenges include access to medicines in vulnerable groups, increasing costs and demand for new medicines, access to prescribers, budgetary constraints, cultural and health literacy, patient affordability and evidence requirement for gaining subsidy for medicines.


The effects of restaurant menu calorie labeling on hypothetical meal choices of females with disordered eating.
Concerns have been raised that obesity public policy measures may have harmful effects on individuals with eating disorders. However, little research has investigated this topic. We examined the impact of a popular obesity public policy, menu calorie labeling, on hypothetical food choices of women with disordered eating. Seven hundred sixteen adult females completed an online survey in which they were randomly assigned to receive a restaurant menu with or without calorie information listed. Participants selected foods representative of a meal they would choose to consume and answered questions on restaurant ordering and menu labeling. Participants completed the Eating Disorder Examination Questionnaire (Fairburn & Beglin, ) to assess global eating pathology. Diagnoses of anorexia nervosa (AN), bulimia nervosa (BN), and binge-eating disorder (BED) were also derived from this measure. Generalized linear modeling examined the impact of menu label condition, disordered eating, and the menu label by disordered eating interaction on hypothetical food selection and related variables. When disordered eating was examined continuously, menu labeling did not differentially affect food selections of those with elevated disordered eating (p = .45). However, when examined by eating disorder diagnosis, participants with AN or BN ordered significantly fewer (p < .001) and participants with BED ordered significantly more (p = .001) calories in the menu label versus no label condition. Menu labeling may decrease the calories ordered among individuals with AN or BN and increase calories ordered among individuals with BED.


A comparison of mechanisms proposed for the conversion of mitomycins into mitosenes.
Two mechanisms proposed for the acid-catalyzed conversions of mitomycins into mitosenes were investigated by deuterium incorporation methods. Four different mitomycins, an aziridinomitosene, and an N-acetylmitomycin all underwent the conversion in acetic acid-d with no incorporation of deuterium at C-1. This evidence suggests that the mechanism based on initial elimination of the elements of methanol to give an aziridinomitosene is more likely the correct one. The products of these reactions had considerable variation in the ratios of cis to trans isomers: 7-aminomitosanes gave a predominance of trans and 7-methoxymitosanes gave a predominance of cis. Treatment of mitomycin C with DCl in D2O gave predominantly cis product with about 45% deuterium exchange at C-1. The isomeric 2,7-diamino-1-methoxymitosenes previously obtained by treating mitomycin C in methanol containing acetic acid were found to have stereochemistry opposite to that originally assigned by us.


Epidemiological, clinical and pathological features of primary cardiac hemangiosarcoma in dogs: a review of 51 cases.
In the study presented here, we aimed to describe the epidemiological, clinical and pathological findings of 51 canine cases with histologically-verified diagnoses of primary cardiac hemangiosarcoma (HSA). The medical data for each dog, including signalment, presenting complaints, physical examination findings, results of various diagnostic testing performed and method of treatment, were checked. In addition, all 51 cases were re-examined pathologically. The tumor occurred most frequently in older Golden Retrievers, followed by Maltese dogs and Miniature Dachshunds. Mass lesions of HSA were found more commonly in the right auricle (RAu) (25/51) and right atrium (RA) (21/51), and the RA masses were significantly (P<0.001) larger than the RAu masses. The echocardiographic detection rate of masses in the RAu group (60%; 15/25) was significantly lower than that in the RA group (95%; 20/21). Survival time was significantly (P<0.05) longer for 5 dogs that received adjuvant chemotherapy after tumor resection than for 12 dogs that did not. In this series, the Maltese (9/51) and Miniature Dachshund (7/51), as well as the Golden Retriever, were represented more frequently than other breeds. The lower echocardiographic detection rate of RAu masses compared with RA masses may be related to tumor size and/or location. The significantly longer survival time for dogs receiving adjuvant chemotherapy indicates that postoperative chemotherapy could be useful for dogs with cardiac HSA.


[Effect of crystallinity on the rate of solution of oxytetracycline hydrochloride].
The crystal structure of 2 forms of oxytetracycline hydrochloride prepared with the method of crystallization from various solvents was studied. It was shown that these forms were 2 polymorphic modifications of the antibiotic with different crystal structures. The modification prepared with the method of crystallization from methanol was characterized by better dissolution in water. Its dissolution rate constant was 3 times as higher which makes this crystalline modification of oxytetracycline hydrochloride preferable for preparation of the antibiotic pharmaceutical forms with high bioavailability levels.


Anti-disialosyl antibodies mediate selective neuronal or Schwann cell injury at mouse neuromuscular junctions.
The human paralytic neuropathy, Miller Fisher syndrome (MFS) is associated with autoantibodies specific for disialosyl epitopes on gangliosides GQ1b, GT1a, and GD3. Since these gangliosides are enriched in synaptic membranes, anti-ganglioside antibodies may target neuromuscular junctions (NMJs), thereby contributing to disease symptoms. We have shown previously that at murine NMJs, anti-disialosyl antibodies induce an alpha-latrotoxin-like effect, electrophysiologically characterized by transient massive increase of spontaneous neurotransmitter release followed by block of evoked release, resulting in paralysis of the muscle preparation. Morphologically, motor nerve terminal damage, as well as perisynaptic Schwann cell (pSC) death is observed. The relative contributions of neuronal and pSC injury to the paralytic effect and subsequent repair are unknown. In this study, we have examined the ability of subsets of anti-disialosyl antibodies to discriminate between the neuronal and glial elements of the NMJ and thereby induce either neuronal injury or pSC death. Most antibodies reactive with GD3 induced pSC death, whereas antibody reactivity with GT1a correlated with the extent of nerve terminal injury. Motor nerve terminal injury resulted in massive uncontrolled exocytosis with paralysis. However, pSC ablation induced no acute (within 1 h) electrophysiological or morphological changes to the underlying nerve terminal. These data suggest that at mammalian NMJs, acute pSC injury or ablation has no major deleterious influence on synapse function. Our studies provide evidence for highly selective targeting of mammalian NMJ membranes, based on ganglioside composition, that can be exploited for examining axonal-glial interactions both in disease states and in normal NMJ homeostasis.


Viral persistence redirects CD4 T cell differentiation toward T follicular helper cells.
CD4 T cell responses are crucial to prevent and control viral infection; however, virus-specific CD4 T cell activity is considered to be rapidly lost during many persistent viral infections. This is largely caused by the fact that during viral persistence CD4 T cells do not produce the classical Th1 cytokines associated with control of acute viral infections. Considering that CD4 T cell help is critical for both CD8 T cell and B cell functions, it is unclear how CD4 T cells can lose responsiveness but continue to sustain long-term control of persistent viral replication. We now demonstrate that CD4 T cell function is not extinguished as a result of viral persistence. Instead, viral persistence and prolonged T cell receptor stimulation progressively redirects CD4 T cell development away from the Th1 response induced during an acute infection toward T follicular helper cells. Importantly, this sustained CD4 T cell functionality is critical to maintain immunity and ultimately aid in the control of persistent viral infection.


Dynamic measurement of the optical properties of bovine enamel demineralization models using four-dimensional optical coherence tomography.
Dental enamel mineral loss is multifactorial and is consequently explored using a variety of in vitro models. Important factors include the presence of acidic pH and its specific ionic composition, which can both influence lesion characteristics. Optical coherence tomography (OCT) has been demonstrated as a promising tool for studying dental enamel demineralization. However, OCT-based characterization and comparison of demineralization model dynamics are challenging without a consistent experimental environment. Therefore, an automated four-dimensional OCT system was integrated with a multispecimen flow cell to measure and compare the optical properties of subsurface enamel demineralization in different models. This configuration was entirely automated, thus mitigating any need to disturb the specimens and ensuring spatial registration of OCT image volumes at multiple time points. Twelve bovine enamel disks were divided equally among three model groups. The model demineralization solutions were citric acid (pH 3.8), acetic acid (pH 4.0), and acetic acid with added calcium and phosphate (pH 4.4). Bovine specimens were exposed to the solution continuously for 48 h. Three-dimensional OCT data were obtained automatically from each specimen at a minimum of 1-h intervals from the same location within each specimen. Lesion dynamics were measured in terms of the depth below the surface to which the lesion extended and the attenuation coefficient. The net loss of surface enamel was also measured for comparison. Similarities between the dynamics of each model were observed, although there were also distinct characteristic differences. Notably, the attenuation coefficients showed a systematic offset and temporal shift with respect to the different models. Furthermore, the lesion depth curves displayed a discontinuous increase several hours after the initial acid challenge. This work demonstrated the capability of OCT to distinguish between different enamel demineralization models by making dynamic quantitative measurements of lesion properties. This has important implications for future applications in clinical dentistry.


Recognition of deglycosylated larval proteins of Gnathostoma spinigerum by a monoclonal antibody and human gnathostomiasis antiserum.
The study on the recognition of 35S-labelled somatic antigens of Gnathostoma spinigerum advanced third-stage larva (aL3) has revealed that the mAb GN6/24 immunoprecipitated 26- and 24-kDa proteins from the undigested and N-glycosidase F-digested larval extracts, respectively. The recognition of the deglycosylated form of the glycoprotein indicated that the mAb reacted with the peptide epitope on the 26-kDa protein. Human gnathostomiasis antiserum immunoprecipitated most of the N-glycosidase F-digested larval proteins including the deglycosylated 26-kDa protein.


The role of the sex-determining region of the Y chromosome (SRY) in the etiology of 46,XX true hermaphroditism.
The syndrome of 46,XX true hermaphroditism is a clinical condition in which both ovarian and testicular tissue are found in one individual. Both Mullerian and Wolffian structures are usually present, and external genitalia are often ambiguous. Two alternative mechanisms have been proposed to explain the development of testicular tissue in these subjects: (1) translocation of chromosomal material encoding the testicular determination factor (TDF) from the Y to the X chromosome or to an autosome, or (2) an autosomal dominant mutation that permits testicular determination in the absence of TDF. We have investigated five subjects with 46,XX true hermaphroditism. Four individuals had a normal 46,XX karyotype; one subject (307) had an apparent terminal deletion of the short arm of one X chromosome. Genomic DNA was isolated from these individuals and subjected to Southern blot analysis. Only subject 307 had Y chromosomal sequences that included the pseudoautosomal boundary, SRY (sex-determining region of Y), ZFY (Y gene encoding a zinc finger protein), and DXYS5 (an anonymous locus on the distal short arm of Y) but lacked sequences for DYZ5 (proximal short arm of Y) and for the long arm probes DYZ1 and DYZ2. The genomic DNA of the other four subjects lacked detectable Y chromosomal sequences when assayed either by Southern blotting or after polymerase chain reaction amplification. Our data demonstrate that 46,XX true hermaphroditism is a genetically heterogeneous condition, some subjects having TDF sequences but most not. The 46,XX subjects without SRY may have a mutation of an autosomal gene that permits testicular determination in the absence of TDF.


Calculating effective dose on a cone beam computed tomography device: 3D Accuitomo and 3D Accuitomo FPD.
This study evaluates two methods for calculating effective dose, CT dose index (CTDI) and dose-area product (DAP) for a cone beam CT (CBCT) device: 3D Accuitomo at field size 30x40 mm and 3D Accuitomo FPD at field sizes 40x40 mm and 60x60 mm. Furthermore, the effective dose of three commonly used examinations in dental radiology was determined. CTDI(100) measurements were performed in a CT head dose phantom with a pencil ionization chamber connected to an electrometer. The rotation centre was placed in the centre of the phantom and also, to simulate a patient examination, in the upper left cuspid region. The DAP value was determined with a plane-parallel transmission ionization chamber connected to an electrometer. A conversion factor of 0.08 mSv per Gy cm(2) was used to determine the effective dose from DAP values. Based on data from 90 patient examinations, DAP and effective dose were determined. CTDI(100) measurements showed an asymmetric dose distribution in the phantom when simulating a patient examination. Hence a correct value of CTDI(w) could not be calculated. The DAP value increased with higher tube current and tube voltage values. The DAP value was also proportional to the field size. The effective dose was found to be 11-77 microSv for the specific examinations. DAP measurement was found to be the best method for determining effective dose for the Accuitomo. Determination of specific conversion factors in dental radiology must, however, be further developed.


Silencing Autophagy-Related Gene 2 (ATG2) Results in Accelerated Senescence and Enhanced Immunity in Soybean.
Autophagy plays a critical role in nutrient recycling and stress adaptations. However, the role of autophagy has not been extensively investigated in crop plants. In this study, soybean autophagy-related gene 2 (GmATG2) was silenced, using virus-induced silencing (VIGS) mediated by Bean pod mottle virus (BPMV). An accelerated senescence phenotype was exclusively observed for the GmATG2-silenced plants under dark conditions. In addition, significantly increased accumulation of both ROS and SA as well as a significantly induced expression of the pathogenesis-related gene 1 (PR1) were also observed on the leaves of the GmATG2-silenced plants, indicating an activated immune response. Consistent with this, GmATG2-silenced plants exhibited a significantly enhanced resistance to Pseudomonas syringae pv. glycinea (Psg) relative to empty vector control plants (BPMV-0). Notably, the activated immunity of the GmATG2-silenced plants was independent of the MAPK signaling pathway. The fact that the accumulation levels of ATG8 protein and poly-ubiquitinated proteins were significantly increased in the dark-treated GmATG2-silenced plants relative to the BPMV-0 plants indicated that the autophagic degradation is compromised in the GmATG2-silenced plants. Together, our results indicated that silencing GmATG2 compromises the autophagy pathway, and the autophagy pathway is conserved in different plant species.


The possible role of NS3 protease activity of hepatitis C virus on fibrogenesis and miR-122 expression in hepatic stellate cells.
The various roles of hepatitis C virus (HCV) NS3 protein in viral pathogenesis are emphasized, especially in the progression of fibrosis and tumors. The levels of miR-122 have been widely accepted as a critical factor in viral pathogenesis and disease progression. However, the possible correlation between miR-122 levels and fibrosis state has been less investigated. Therefore, in this study, plasmids expressing protease competent and protease mutated non-structural proteins 3 (NS3) were transfected into LX-2 cell line. Subsequently, the total RNA was extracted and real-time PCR was performed to measure the expression level of miR-122, collagen type 1 alpha 1 (COL1A1), alpha smooth muscle actin (α-SMA), and tissue inhibitor of metaloproteinase 1 (TIMP-1). Moreover, the transforming growth factor beta (TGF-β) levels in the supernatants of transfected cells were evaluated by ELISA. The gene expression analysis of fibrotic genes and TGF-β cytokine in LX-2 cells showed that protease competent NS3 had a significant fibrogenic impact when compared to protease defective NS3 or GFP control plasmids (P <0.001). The results also demonstrated that the expression of miR-122 was downregulated in both versions of the cells transfected with NS3 plasmids (P <0.01) irrespective of protease function. These results suggested that the protease function of NS3 protein is a crucial factor for the induction of hepatic fibrosis but it doesn't play a complete role in the expression of miR-122.


Homoglutathione confers tolerance to acifluorfen in transgenic tobacco plants expressing soybean homoglutathione synthetase.
Homoglutathione (hGSH), which is present in some leguminous plants, is preferred over GSH in in vitro conjugation of acifluorfen and fomesafen by glutathione S-transferase. To investigate the function of hGSH in in vivo detoxification of xenobiotics, we evaluated herbicide tolerance of transgenic tobacco plants expressing soybean homoglutathione synthetase in the cytosol or chloroplasts. Transgenic plants synthesizing hGSH in the cytosol were more tolerant to acifluorfen than wild-type plants, whereas enhanced tolerance to fomesafen was not observed. Transgenic plants synthesizing hGSH in the chloroplasts showed no enhanced tolerance to acifluorfen or fomesafen.


[The effectiveness of intravaginal estriol tablet administration in women with urge incontinence].
In postmenopausal patients, stress and urge incontinence often occur as a consequence of oestrogen deficiency. We performed a comparative study in a prospectively randomised and placebo controlled group of women with urge incontinence to investigate different dosages of intravaginally applied oestriol. Clinical and urodynamic parameters were also compared. 15 women (Group I) received 1 mg oestriol applied daily intravaginally over 3 weeks; 15 women (Group II) received a daily dosage of 3 mg and 10 women received a placebo. A complete clinical and urodynamic evaluation was carried out twice at a 4-week interval. The women receiving a daily dose of 3 mg oestriol applied intravaginally demonstrated a significant improvement of the parameters strong desire to void, pollakisuria, and nycturia. No improvement was seen in patients with motoric urge incontinence.


The association between the level of adherence to the Mediterranean diet and successful aging: An analysis of the ATTICA and MEDIS (MEDiterranean Islands Study) epidemiological studies.
The aim of the present work was to evaluate the association between the level of adherence to the Mediterranean diet and successful aging, taking into consideration the overall dietary pattern, rather than individual foods, among middle-aged and older individuals. A harmonized dataset of middle-aged and older participants (>50 years old) from the ATTICA (n = 1,128) and MEDIS (n = 2,221) population-based cross-sectional studies was used. Socio-demographic, anthropometric, clinical and lifestyle characteristics were measured in both studies and harmonized using standardized procedures. Level of adherence to the Mediterranean diet was evaluated using the MedDietScore (range 0-55, "Low" adherence <34, "High">38). Successful aging was evaluated using the validated successful aging index (SAI, range 0-10) comprising of health-related, social, lifestyle and clinical characteristics. Adherence to the Mediterranean diet was positively associated with SAI (b ± SE "High" vs. "Low": 1.094 ± 0.130, p < 0.001 and "Moderate" vs "Low": 0.476 ± 0.156, p = 0.003). Higher adherence to the Mediterranean diet was independently associated with a higher level of successful aging. Mediterranean diet or dietary patterns that are close to this pattern, should be actively promoted and encouraged to middle aged and older people to achieve successful aging.


Ultrasoft Alginate Hydrogels Support Long-Term Three-Dimensional Functional Neuronal Networks.
Neuron development and function are exquisitely sensitive to the mechanical properties of their surroundings. Three-dimensional (3D) cultures are therefore being explored as they better mimic the features of the native extracellular matrix. Limitations of existing 3D culture models include poorly defined composition, rapid degradation, and suboptimal biocompatibility. Here we show that ionically cross-linked ultrasoft hydrogels made from unmodified alginate can potently promote neuritogenesis. Alginate hydrogels were characterized mechanically and a remarkable range of stiffness (10-4000 Pa) could be produced by varying the macromer content (0.1-0.4% w/v) and CaCl2 concentration. Dissociated rat embryonic cortical neurons encapsulated within the softest of the hydrogels (0.1% w/v, 10 mM CaCl2) showed excellent viability, extensive formation of axons and dendrites, and long-term activity as determined by calcium imaging. In conclusion, alginate is an off-the-shelf, easy to handle, and inexpensive material, which can be used to make ultrasoft hydrogels for the formation of stable and functional 3D neuronal networks. This 3D culture system could have important applications in neuropharmacology, toxicology, and regenerative medicine.


Erratum.
In the report "Positron emission tomography reveals elevated D(2) dopamine receptors in drugnaive schizophrenics" by D. F. Wong et al. (19 Dec., p. 1558), reference 8 should have read, "D. F. Wong et al., Science 226, 1393 (1984); D. F. Wong et al., J. Cereb. Blood Flow Metab. 6, 147 ( 1986).


[Blood donation after reaching 65 years of age].
European regulations for blood donation recommend a maximum donor age of 65 years. On the other hand, the percentage of the population in this age group is rapidly increasing in Western countries, and in autologous blood donation programs this limitation has already been abandoned. In a prospective study we examined blood donation in elderly donors (18 male and 5 female; mean age 65 years, range 64-69) in comparison to a younger control group (15/9; 58, 55-63). All donors were regular blood donors and had donated for at least 3 years. We investigated the exercise capacity before and after donation of 450 ml whole blood by examination of the physical working capacity (PWC) at heart rates of 110/min and 130/min through treadmill exercise testing and determined the blood viscosity. Additionally, whole-blood count, hemoglobin, plasma-ferritin levels and total iron binding capacity were measured immediately after donation and on days 7, 28 and 49. We found a decrease in whole-blood viscosity and a moderate increase in PWC at heart rates of 110/min and 130/min after donation in both groups. Red cell count and values of hemoglobin and ferritin were significantly lower in both groups after donation and returned to pre-donation values by day 49 in the younger control group. We detected no deterioration in exercise capacity after whole-blood donation in elderly blood donors over 65 years when compared with a younger control group. We suggest that blood donation in otherwise healthy persons aged over 65 years should be accepted.


Nerve growth factor and norepinephrine concentrations in weight-bearing and non-weight-bearing bones of euthyroid and hyperthyroid rats.
Thyroid hormone, nerve growth factor (NGF) and norepinephrine (NE) and weight-bearing affect bone metabolism, yet interactions between these factors and osseous tissue have not been investigated. Therefore, the aims of the study were to measure NGF and NE concentrations in weight-bearing and non-weight-bearing bones from euthyroid (control) and hyperthyroid (HT) rats. Hyperthyroidism was induced by oral intake of triiodothyronine (90 mg/kg/day) for 21 days. Histomorphometry on distal femurs verified significant trabecular bone loss in HT rats compared to euthyroid animals. NGF concentrations were assayed via ELISA, whilst NE concentrations were measured via HPLC and ECD. In euthyroid rats: (i) the concentration of NGF in ribs (914 ng/g) was almost 3-fold greater than in femurs (326 ng/g wet weight of tissue) (ii) the concentrations of NE in ribs (74.7 ng/g) and calvaria (87.4 ng/g) were 2.5-3.5-fold greater than either femurs (24.0 ng/g) or tibiae (30.5 ng/g) and (iii) NE concentrations were comparable between ribs (74.7 ng/g) and calvaria (87.4 ng/g) and similar between tibiae (30.5 ng/g) and femurs (24.0 ng/g). In HT rats: (i) the concentration of NGF in ribs (1802 ng/g) was 4-fold greater than in femurs (402 ng/g) (ii) NE concentrations in ribs (23.3 ng/g) and calvaria (13.6 ng/g) were 4.5-fold and 2.6-fold greater respectively than in tibiae (5.2 ng/g), while ribs had almost a 2-fold higher concentration of NE than calvaria. In HT rats compared to euthyroid animals: (i) NGF concentrations almost doubled in ribs but there was little change in the NGF concentration in femurs (ii) there was a reduction in NE concentrations in calvaria by 84%, in ribs by 69% in tibiae by 83% and 55% in femur (NS). (i) Non-weight-bearing is associated with higher concentrations of NGF and NE than weight-bearing in bones in euthyroid and HT rats; (ii) Hyperthyroidism exerts opposite effects on NGF and NE in bone and (iii) Hyperthyroidism interacts with weight-bearing to determine NGF and NE concentrations in bone. Therefore, the influence of thyroid hormone on NGF and NE in bone may need to be taken into account when considering the action of thyroid hormone on bone in either euthyroid or hyperthyroid states.


Examination of virus-infected cultured cells by scanning electron microscopy.
The scanning electron microscope (SEM) was used to detect changes in morphology of BSC-1 cells after infection with vesicular stomatitis virus (VSV) or herpes simplex virus. The morphological changes of the infected cells were related to the length of time of infection and to the virus used. Extensive alteration to the cytoplasm could be seen 24 and 48 hr after infection with 10 and 320 TCID(50) of VSV. Within 24 hr after infection with 1 TCID(50) of herpes simplex, a few nuclei were swollen. However, 72 hr after infection with 100 TCID(50) of herpesvirus, many nuclei were swollen and appeared in large aggregates, probably representing formation of a polykaryocyte. Corresponding samples stained with May-Grunwald-Giemsa were observed in the light microscope and morphological changes were compared to those seen with the SEM.


Assessment of the German and Italian Stress Cardiomyopathy Score for Risk Stratification for In-hospital Complications in Patients With Takotsubo Syndrome.
Takotsubo syndrome (TTS) is an acute, reversible heart failure syndrome featured by significant rates of in-hospital complications. There is a lack of data for risk stratification during hospitalization. To derive a simple clinical score for risk prediction of in-hospital complications among patients with TTS. In this prognostic study, 1007 consecutive patients were enrolled in the German and Italian Stress Cardiomyopathy (GEIST) registry from July 1, 2007, through December 31, 2017, and identified as the derivation cohort; 946 patients were enrolled in the Spanish Registry for Takotsubo Cardiomyopathy (RETAKO) as the external score validation. An admission risk score was developed using a stepwise multivariable regression analysis from 2 registries. Data analysis was performed from March 1, 2018, through July 31, 2018. In-hospital complications were defined as death, pulmonary edema, need for invasive ventilation, and cardiogenic shock. Four variables were identified as independent predictors of in-hospital complications and were used for the score: male sex, history of neurologic disorder, right ventricular involvement, and left ventricular ejection fraction (LVEF). Of the 1007 patients enrolled in the GEIST registry, 107 (10.6%) were male, with mean (SD) age of 69.8 (11.4) years. Overall rate of in-hospital complications was 23.3% (235 of 1007) (death, 4.0%; pulmonary edema, 5.8%; invasive ventilation, 6.4%; and cardiogenic shock, 9.1%). The GEIST prognosis score was derived by providing 20 points each for male sex and history of neurologic disorders and 30 points for right ventricular involvement and then subtracting the value in percent of LVEF (decimal values between 0.15 and 0.70). Score accuracy on area under the receiver operating characteristic curve analysis was 0.71, with a negative predictive power of 87% with scores less than 20. External validation in the RETAKO population (124 [13.1%] male; mean [SD] age, 69.5 [14.9] years) revealed an area under the curve of 0.73 (P = .46 vs GEIST derivation cohort). Stratification into 3 risk groups (<20, 20-40, and >40 points) classified 316 patients (40.9%) as having low risk; 342 (44.3%) as having intermediate risk, and 114 (14.8%) as having high risk of complications. The observed in-hospital complication rates were 12.7% for low-risk patients, 23.4% for intermediate-risk patients, and 58.8% for high-risk patients (P < .001 for trend). After 2.6 years of follow-up, patients with in-hospital complications had significantly higher rates of mortality than those without complications (40% vs 10%, P = .01). The GEIST prognostic score may be useful in early risk stratification for TTS. High-risk patients with TTS may require an intensive care unit stay, and low-risk patients with TTS could be discharged within a few days. In-hospital complications in patients with TTS may be associated with increased risk of long-term mortality.


Triple valve surgery through a less invasive approach: early and mid-term results.
A partial upper sternotomy has become established as a less invasive approach mainly for single and double valve surgery. This report evaluates the clinical outcomes of triple valve surgery performed through a partial upper sternotomy. We reviewed the medical records of 37 consecutive patients (28 men, 76%) who underwent triple valve surgery through a partial upper sternotomy between 2005 and 2015. The patients' mean age was 67 ± 17 years; 27 (73%) were in New York Heart Association Class III or IV. Aortic and mitral valve insufficiency was more common than stenosis. Ninety-three percent of surviving patients were followed for a mean period of 58 ± 24 months. Aortic valve procedures consisted of 24 (65%) replacements and 13 (35%) repairs. The mitral valve was repaired in 28 (76%) patients, whereas tricuspid valve repair was feasible in all patients. No conversion to full sternotomy was necessary. Myocardial infarction was not observed. Chest tube drainage was 330 ± 190 ml, and 4 patients required reopening for bleeding (1, 3%) or tamponade (3, 8%). One stroke was observed due to heparin-induced thrombocytopaenia after initial unremarkable neurological recovery. Early mortality included 5 (13.5%) patients. Actuarial survival at 5 years was 52 ± 10%. A partial upper sternotomy provides adequate exposure to all heart valves. We did not experience technical limitations with this approach. Wound dehiscence, postoperative bleeding, intensive care unit and hospital stay and early deaths were low compared to data from other published series of triple valve surgery through a full median sternotomy. Early and mid-term outcomes were not adversely affected by this less invasive approach.


Synthetic peptides corresponding to third hypervariable region of human monoclonal IgM rheumatoid factor heavy chains define an immunodominant idiotype.
Synthetic peptides corresponding to eight individual heavy chain complementarity-determining regions (CDR) of three human monoclonal IgM anti-IgG (rheumatoid factor [RF]) paraproteins elicited rabbit antibodies with markedly different properties. All antisera recognized the immunizing peptide, and several reacted with the isolated IgM heavy chain on immunoblots. However, only the antisera against peptides representing the third CDR bound consistently and specifically to the intact IgM-RF molecule. These data indicate that the third CDR of human mu chains comprises an immunodominant idiotype, and suggest that the D gene segment may be especially important in creating idiotypic diversity. Synthetic peptides corresponding to the third heavy chain CDR of human paraproteins may be clinically useful for the specific induction of antiidiotypic antibodies.


In vivo gene transfer to the human biliary tract.
The human biliary tract offers an excellent model for gene transfer studies for a variety of diseases localized to the liver. The aim of this study was to determine if a viable liver might be employed to study viral transfection of the human biliary system in order to mimic in vivo human experiments. Using a normal human liver initially procured for transplantation, but subsequently found unsuitable, and with an intact biliary tree, the hepatic vascular supply was accessed for continuous perfusion. The common and left hepatic biliary system was isolated by balloon catheterization. A replication defective adenoviral vector containing the Escherichia coli beta-galactosidase (lac Z) reporter gene (AdCMVLacZ) was injected into the catheter-isolated left and common bile duct lumen. Viral exposure to the right duct system was prevented by ligation. The bile duct segments were excised and prepared for enzymatic (X-gal) staining. Intense staining was observed in the biliary epithelium exposed to the adenoviral vector. No evidence of beta-galactosidase staining was noted in the unexposed biliary mucosa. We report direct transfection of biliary epithelial cells from normal human liver with a recombinant adenovirus. Our data suggest potential therapeutic applications for gene therapy of hepatobiliary disorders.


Runge-Kutta neural network for identification of dynamical systems in high accuracy.
This paper proposes Runge-Kutta neural networks (RKNNs) for identification of unknown dynamical systems described by ordinary differential equations (i.e., ordinary differential equation or ODE systems) with high accuracy. These networks are constructed according to the Runge-Kutta approximation method. The main attraction of the RKNNs is that they precisely estimate the changing rates of system states (i.e., the right-hand side of the ODE x =f(x)) directly in their subnetworks based on the space-domain interpolation within one sampling interval such that they can do long-term prediction of system state trajectories. We show theoretically the superior generalization and long-term prediction capability of the RKNNs over the normal neural networks. Two types of learning algorithms are investigated for the RKNNs, gradient-and nonlinear recursive least-squares-based algorithms. Convergence analysis of the learning algorithms is done theoretically. Computer simulations demonstrate the proved properties of the RKNNs.


Requirements for the nuclear export of the small ribosomal subunit.
Eukaryotic ribosome biogenesis requires multiple steps of nuclear transport because ribosomes are assembled in the nucleus while protein synthesis occurs in the cytoplasm. Using an in situ RNA localization assay in the yeast Saccharomyces cerevisiae, we determined that efficient nuclear export of the small ribosomal subunit requires Yrb2, a factor involved in Crm1-mediated export. Furthermore, in cells lacking YRB2, the stability and abundance of the small ribosomal subunit is decreased in comparison with the large ribosomal subunit. To identify additional factors affecting small subunit export, we performed a large-scale screen of temperature-sensitive mutants. We isolated new alleles of several nucleoporins and Ran-GTPase regulators. Together with further analysis of existing mutants, we show that nucleoporins previously shown to be defective in ribosomal assembly are also defective in export of the small ribosomal subunit.


Alkaloid Lindoldhamine Inhibits Acid-Sensing Ion Channel 1a and Reveals Anti-Inflammatory Properties.
Acid-sensing ion channels (ASICs), which are present in almost all types of neurons, play an important role in physiological and pathological processes. The ASIC1a subtype is the most sensitive channel to the medium's acidification, and it plays an important role in the excitation of neurons in the central nervous system. Ligands of the ASIC1a channel are of great interest, both fundamentally and pharmaceutically. Using a two-electrode voltage-clamp electrophysiological approach, we characterized lindoldhamine (a bisbenzylisoquinoline alkaloid extracted from the leaves of Laurus nobilis L.) as a novel inhibitor of the ASIC1a channel. Lindoldhamine significantly inhibited the ASIC1a channel's response to physiologically-relevant stimuli of pH 6.5-6.85 with IC50 range 150-9 μM, but produced only partial inhibition of that response to more acidic stimuli. In mice, the intravenous administration of lindoldhamine at a dose of 1 mg/kg significantly reversed complete Freund's adjuvant-induced thermal hyperalgesia and inflammation; however, this administration did not affect the pain response to an intraperitoneal injection of acetic acid (which correlated well with the function of ASIC1a in the peripheral nervous system). Thus, we describe lindoldhamine as a novel antagonist of the ASIC1a channel that could provide new approaches to drug design and structural studies regarding the determinants of ASIC1a activation.


Economics of zoonoses surveillance in a 'One Health' context: an assessment of Campylobacter surveillance in Switzerland.
Cross-sectorial surveillance and general collaboration between the animal and the public health sectors are increasingly recognized as needed to better manage the impacts of zoonoses. From 2009, the Swiss established a Campylobacter mitigation system that includes human and poultry surveillance data-sharing within a multi-sectorial platform, in a 'One Health' approach. The objective of this study was to explore the economics of this cross-sectorial approach, including surveillance and triggered interventions. Costs and benefits of the One Health and of the uni-sectorial approach to Campylobacter surveillance were identified using an economic assessment framework developed earlier. Cost information of surveillance activities and interventions was gathered and disability-adjusted life years (DALYs) associated with the disease estimated for 2008 and 2013. In the first 5 years of this One Health approach to Campylobacter mitigation, surveillance contributed with information mainly used to perform risk assessments, monitor trends and shape research efforts on Campylobacter. There was an increase in costs associated with the mitigation activities following integration, due mainly to the allocation of additional resources to research and implementation of poultry surveillance. The overall burden of campylobacteriosis increased by 3·4-8·8% to 1751-2852 DALYs in 2013. In the timing of the analysis, added value associated with this cross-sectorial approach to surveillance of Campylobacter in the country was likely generated through non-measurable benefits such as intellectual capital and social capital.


[Metabolic state of the other eye in post-traumatic uveitis].
Biochemical analysis of the paired eye lacrimal fluid lipid peroxides (diene conjugates, Schiff's bases, TBA-active products), antioxidant enzymes (catalase, superoxide dismutase), dehydrogenases (lactate and malate dehydrogenases), and lactic and pyruvic acids was carried out during the early and remote posttraumatic period in 468 patients after perforating injuries to the eye. Signs of activation of lipid peroxidation, antioxidants, dehydrogenases, and of glycolysis intensification were detected. These changes corresponded to the results of analysis of lacrimal fluid of the injured eye and depended on the severity of injury, time elapsed after it, and severity and course of posttraumatic uveitis. The author claims that the detected metabolic shifts in the lacrimal fluid of paired eye are involved in the pathogenesis of posttraumatic uveitis.


SMAD4: A critical regulator of cardiac neural crest cell fate and vascular smooth muscle development.
During embryogenesis, cardiac neural crest-derived cells (NCs) migrate into the pharyngeal arches and give rise to the vascular smooth muscle cells (vSMCs) of the pharyngeal arch arteries (PAAs). vSMCs are critical for the remodeling of the PAAs into their final adult configuration, giving rise to the aortic arch and its arteries (AAAs). We investigated the role of SMAD4 in NC-to-vSMC differentiation using lineage-specific inducible mouse strains. We found that the expression of SMAD4 in the NC is indelible for regulating the survival of cardiac NCs. Although the ablation of SMAD4 at E9.5 in the NC lineage led to a near-complete absence of NCs in the pharyngeal arches, PAAs became invested with vSMCs derived from a compensatory source. Analysis of AAA development at E16.5 showed that the alternative vSMC source compensated for the lack of NC-derived vSMCs and rescued AAA morphogenesis. Our studies uncovered the requisite role of SMAD4 in the contribution of the NC to the pharyngeal arch mesenchyme. We found that in the absence of SMAD4+ NCs, vSMCs around the PAAs arose from a different progenitor source, rescuing AAA morphogenesis. These findings shed light on the remarkable plasticity of developmental mechanisms governing AAA development.


Channelized model observer for the detection and estimation of signals with unknown amplitude, orientation, and size.
As a task-based approach for medical image quality assessment, model observers (MOs) have been proposed as surrogates for human observers. While most MOs treat only signal-known-exactly tasks, there are few studies on signal-known-statistically (SKS) MOs, which are clinically more relevant. In this paper, we present a new SKS MO named channelized joint detection and estimation observer (CJO), capable of detecting and estimating signals with unknown amplitude, orientation, and size. We evaluate its estimation and detection performance using both synthesized (correlated Gaussian) backgrounds and real clinical (magnetic resonance) backgrounds. The results suggest that the CJO has good performance in the SKS detection-estimation task.


Bundling of collagen fibrils influences osteocyte network formation during bone modeling.
Osteocytes form a cellular network by gap junctions between their cell processes. This network is important since intercellular communication via the network is essential for bone metabolism. However, the factors that influence the formation of this osteocyte network remain unknown. As the early stage of osteocyte network formation occurs on the bone surface, we observed a newly formed trabecular bone surface by orthogonal focused ion beam-scanning electron microscopy. The embedding late osteoblast processes tended to avoid bundled collagen fibrils and elongate into sparse collagen fibrils. Then, we examined whether the inhibition of bundling of collagen fibrils using a potent lysyl oxidase inhibitor, β-aminopropionitrile (BAPN) changed the cellular network of the chick calvaria. The osteocyte shape of the control group was spindle-shape, while that of the BAPN group was sphere-shaped. In addition, the osteocyte processes of the control group were elongated vertically to the long axis of the cell body, whereas the osteocyte processes of the BAPN group were elongated radially. Therefore, it was suggested that the bundling of collagen fibrils influences normal osteocyte network formation during bone modeling.


Manganese inhibits poly(ADP-ribosyl)ation in human cells: a possible mechanism behind manganese-induced toxicity?
For humans manganese is both an essential trace element and, at higher doses, a toxic metal. Due to the ubiquitous occurrence of manganese in foodstuff, in industrial countries daily dietary uptake is higher as compared to the estimated daily requirement. Therefore manganese deficiency is extremely rare. In contrast chronic manganese toxicity, affecting primarily the central nervous system, is more prevalent. Thus manganese occupational and dietary overexposure has been shown to cause progressive, permanent, neurodegenerative damage, resulting in syndromes similar to idiopathic Parkinson's disease. To date modes of manganese neurotoxic action are poorly understood and in most studies oxidative stress is postulated as the underlying mechanism. The present study searched on the cellular level for a molecular mechanism behind manganese-induced neurotoxicity and investigated bioavailability, cytotoxicity and genotoxicity of MnCl(2), as well as its impact on the DNA damage response in human cells (HeLa S3) in culture. Whereas up to 10 µM MnCl(2) showed no induction of DNA strand breaks after 24 h incubation, manganese strongly inhibited H(2)O(2)-stimulated poly(ADP-ribosyl)ation at low, completely non-cytotoxic, for certain human exposure, relevant concentrations starting at 1 µM. Thereby inhibition of this essential DNA damage response signalling reaction was not due to a reduced gene expression or protein level of the responsible polymerase PARP-1. Taken together, the results indicate that manganese, under conditions of either overload due to high exposure or disturbed homeostasis, can disturb the cellular response to DNA strand breaks, which has been shown before (S. Katyal and P. J. McKinnon, Mech. Ageing Dev., 2008, 129, 483-491) to result in neurological diseases.


Cost-effectiveness of rivaroxaban versus apixaban for the initial treatment of venous thromboembolism and extended prevention of recurrences in the UK.
Aim: To evaluate the relative cost-effectiveness of using rivaroxaban vs apixaban for the initial treatment plus extended prevention of venous thromboembolism (VTE) in the UK. Extended prevention was assessed using a 10-mg rivaroxaban dose, as the 20-mg dose has already been evaluated. Methods: A Markov model compared the health outcomes and costs of treating VTE patient cohorts with either rivaroxaban (15 mg twice daily for 3 weeks, followed by 20 mg once daily for 6 months, then extended prevention with 10 mg once daily) or apixaban (10 mg twice daily for 1 week, followed by 5 mg twice daily for 6 months, then extended prevention with 2.5 mg twice daily) over a lifetime horizon. The model included an initial acute treatment and prevention phase (0-6 months) and an extended prevention phase (6-18 months). Efficacy and safety data were derived from two network meta-analyses. Reference treatment comparators were derived from the EINSTEIN-Pooled study and EINSTEIN-CHOICE trial. Healthcare costs and utility data were derived from published literature. Results: The rivaroxaban regimen was associated with increased quality-adjusted life years (QALYs) and slightly lower total costs compared with apixaban over a lifetime horizon. Deterministic and probabilistic sensitivity analyses demonstrated that rivaroxaban remained a cost-effective alternative to apixaban over a wide range of parameters. Incremental cost-effectiveness ratio estimates were below the £20,000 per QALY threshold in 74.1% of 2,000 model simulations. Scenario analyses further supported that rivaroxaban is a cost-effective alternative to apixaban. Limitations: Clinical and safety inputs were derived from network meta-analysis, which are subject to inherent limitations whereby small differences between study designs may severely impact efficacy and safety outcomes. Furthermore, these inputs were based on data from clinical trials, which may not reflect real-world data. Conclusions: Rivaroxaban was associated with a slightly lower total cost and increased QALYs compared with apixaban for VTE management in the UK over a lifetime horizon.


Reliability, Validity, and Clinical Utility of the Dominic Interactive for Adolescents-RevisedA DSM-5-Based Self-Report Screen for Mental Disorders, Borderline Personality Traits, and Suicidality.
The Dominic Interactive for Adolescents-Revised (DIA-R) is a multimedia self-report screen for 9 mental disorders, borderline personality traits, and suicidality defined by the fifth edition of the Diagnostic and Statistical Manual of Mental Disorders ( DSM-5). This study aimed to examine the reliability and the validity of this instrument. French- and English-speaking adolescents aged 12 to 15 years ( N = 447) were recruited from schools and clinical settings in Montreal and were evaluated twice. The internal consistency was estimated by Cronbach alpha coefficients and the test-retest reliability by intraclass correlation coefficients. Cutoff points on the DIA-R scales were determined by using clinically relevant measures for defining external validation criteria: the Schedule for Affective Disorders and Schizophrenia for School-Aged Children, the Beck Hopelessness Scale, and the Abbreviated-Diagnostic Interview for Borderlines. Receiver operating characteristic (ROC) analyses provided accuracy estimates (area under the ROC curve, sensitivity, specificity, likelihood ratio) to evaluate the ability of the DIA-R scales to predict external criteria. For most of the DIA-R scales, reliability coefficients were excellent or moderate. High or moderate accuracy estimates from ROC analyses demonstrated the ability of the DIA-R thresholds to predict psychopathological conditions. These thresholds were generally capable to discriminate between clinical and school subsamples. However, the validity of the obsessions/compulsions scale was too low. Findings clearly support the reliability and the validity of the DIA-R. This instrument may be useful to assess a wide range of adolescents' mental health problems in the continuum of services. This conclusion applies to all scales, except the obsessions/compulsions one.


Validation of Predicting Hyperglycemic Crisis Death Score: A Risk Stratification Tool for Appropriate Disposition of Hyperglycemic Crisis Patients from the Emergency Department.
Predicting hyperglycemic crisis death (PHD) score is a simple, rapid tool with six independent mortality predictors to calculate 30-day mortality and appropriately dispose patients to Intensive Care Unit (ICU) or ward. This study aimed at validating the efficiency of PHD score as a decision rule for prognosticating 30-day mortality and classifying hyperglycemic crisis patients for appropriate disposition from the emergency department (ED). This is a prospective, observational study done in the ED of a teaching hospital over 14 months. All patients aged >18 years and who met the criteria of hyperglycemic crisis were enrolled. Thirty-day mortality of six independent predictors was the primary end point. Using PHD, risk scores were calculated and patients were disposed as per physician's clinical judgment. Finally, the treating physician's decision and PHD score disposition were compared and the efficiency of PHD in predicting 30-day mortality was analyzed. Multiple logistic regression models were used for analysis. Receiver operating characteristic curve was drawn, and area under the curve along with sensitivity, specificity, positive predictive value, and negative predictive value was analyzed. P < 0.05 was considered statistically significant. A total of 133 patients were included. On applying PHD score, 69, 39, and 25 patients were in the low-, intermediate-, and high-risk groups, respectively, with a mortality rate of 5.8%, 20.5%, and 56%, respectively. On comparing physician disposition with PHD score, an increasing mortality was noticed in ICU, and PHD showed equal weight in risk stratification and appropriate disposition of patients. In adult patients with hyperglycemic crisis, PHD score is validated as a straightforward, prompt tool for predicting 30-day mortality and aids in disposition. The mortality rate in the PHD score Model II was similar to the physician's clinical decision.


Aurora kinase A is required for hematopoiesis but is dispensable for murine megakaryocyte endomitosis and differentiation.
Aurora kinase A (AURKA) is a therapeutic target in acute megakaryocytic leukemia. However, its requirement in normal hematopoiesis and megakaryocyte development has not been extensively characterized. Based on its role as a cell cycle regulator, we predicted that an Aurka deficiency would lead to severe abnormalities in all hematopoietic lineages. Here we reveal that loss of Aurka in hematopoietic cells causes profound cell autonomous defects in the peripheral blood and bone marrow. Surprisingly, in contrast to the survival defects of nearly all hematopoietic lineages, deletion of Aurka was associated with increased differentiation and polyploidization of megakaryocytes both in vivo and in vitro. Furthermore, in contrast to other cell types examined, megakaryocytes continued DNA synthesis after loss of Aurka. Thus, like other cell cycle regulators such as Aurkb and survivin, Aurka is required for hematopoiesis, but is dispensable for megakaryocyte endomitosis. Our work supports a growing body of evidence that the megakaryocyte endomitotic cell cycle differs significantly from the proliferative cell cycle.


A zebrafish model of chordoma initiated by notochord-driven expression of HRASV12.
Chordoma is a malignant tumor thought to arise from remnants of the embryonic notochord, with its origin in the bones of the axial skeleton. Surgical resection is the standard treatment, usually in combination with radiation therapy, but neither chemotherapeutic nor targeted therapeutic approaches have demonstrated success. No animal model and only few chordoma cell lines are available for preclinical drug testing, and, although no druggable genetic drivers have been identified, activation of EGFR and downstream AKT-PI3K pathways have been described. Here, we report a zebrafish model of chordoma, based on stable transgene-driven expression of HRASV12 in notochord cells during development. Extensive intra-notochordal tumor formation is evident within days of transgene expression, ultimately leading to larval death. The zebrafish tumors share characteristics of human chordoma as demonstrated by immunohistochemistry and electron microscopy. The mTORC1 inhibitor rapamycin, which has some demonstrated activity in a chordoma cell line, delays the onset of tumor formation in our zebrafish model, and improves survival of tumor-bearing fish. Consequently, the HRASV12-driven zebrafish model of chordoma could enable high-throughput screening of potential therapeutic agents for the treatment of this refractory cancer.


Resuspension of sediment-associated Escherichia coli in a natural stream.
In this study, a tracer bacteria was used to investigate the resuspension and persistence of sediment-associated bacteria in a small alluvial stream. The study was conducted in Swan Creek, located within the Grand River watershed of Ontario, Canada. A 1.1-m2 section of the bed was seeded with a strain of Escherichia coli resistant to nalidixic acid (E. coli NAR). The survival, transport, and redistribution of the tracer bacteria within a 1.7-km river section downstream of the source cell was assessed for a 2-mo period following the introduction of the tracer bacteria. This study has illustrated that enteric bacteria can survive in bed sediments for up 6 wk and that inactivation of the tracer bacteria resembled typical first-order decay. Critical conditions for resuspension, as well as resuspension rates, of sediment-associated bacteria were determined for several storm events. The critical shear stress for E. coli NAR resuspension in Swan Creek ranged from 1.5 to 1.7 N m(-2), which is comparable with literature values for critical shear stresses for erosion of cohesive sediments. Bacteria resuspension was primarily limited to the rising limb of storm hydrographs implying that a finite supply of sediment-associated bacteria are available for resuspension during individual storm events. The information presented in this paper will further the development of representative microbial water quality models.


Mechanism, regulation, and ecological role of bacterial cyanide biosynthesis.
A few bacterial species are known to produce and excrete hydrogen cyanide (HCN), a potent inhibitor of cytochrome c oxidase and several other metalloenzymes. In the producer strains, HCN does not appear to have a role in primary metabolism and is generally considered a secondary metabolite. HCN synthase of proteobacteria (especially fluorescent pseudomonads) is a membrane-bound flavoenzyme that oxidizes glycine, producing HCN and CO2. The hcnABC structural genes of Pseudomonas fluorescens and P. aeruginosa have sequence similarities with genes encoding various amino acid dehydrogenases/oxidases, in particular with nopaline oxidase of Agrobacterium tumefaciens. Induction of the hcn genes of P. fluorescens by oxygen limitation requires the FNR-like transcriptional regulator ANR, an ANR recognition sequence in the -40 region of the hcn promoter, and nonlimiting amounts of iron. In addition, expression of the hcn genes depends on a regulatory cascade initiated by the GacS/GacA (global control) two-component system. This regulation, which is typical of secondary metabolism, manifests itself during the transition from exponential to stationary growth phase. Cyanide produced by P. fluorescens strain CHA0 has an ecological role in that this metabolite accounts for part of the biocontrol capacity of strain CHA0, which suppresses fungal diseases on plant roots. Cyanide can also be a ligand of hydrogenases in some anaerobic bacteria that have not been described as cyanogenic. However, in this case, as well as in other situations, the physiological function of cyanide is unknown.


Transposition of the carbenicillin-hydrolyzing beta-lactamase gene.
We isolated a new transposon Tn2101, from plasmid Rms433 in Enterobacter cloacae. Tn2101 encoded the formation of type IV (carbenicillin-hydrolyzing) beta-lactamase and multiple resistance to streptomycin, sulfanilamide, spectinomycin, and mercury in addition to ampicillin. Tn2101 was transposable between conjugative (or nonconjugative) plasmids and the host chromosome. Transposition occurred independently of the general recombination ability of the host cell. Tn2101 had a molecular size of 9.5 x 10(6) and contained short inverted repeat terminal sequences.


Effectiveness of antipsychotic drugs in patients with chronic schizophrenia: efficacy, safety and cost outcomes of CATIE and other trials.
After the introduction of clozapine in 1990, second-generation-or atypical-antipsychotics were thought to be more effective, safer, and less costly than first-generation-or typical-antipsychotics. However, only limited evidence on the efficacy and safety of atypicals was available. Because of this knowledge gap, the National Institute of Mental Health sponsored the Clinical Antipsychotic Trials of Intervention Effectiveness (CATIE) to compare first- and second-generation antipsychotics. The CATIE trial found that antipsychotic drug treatments are generally effective overall but have various limitations as reflected by high rates of discontinuation due to both efficacy and tolerability problems. In addition, the trial found that conventional agents with intermediate potency were comparably effective with atypical agents.


[Pharmacoradiography of the gastro intestinal tract (author's transl)].
Pharmacoradiography of the digestive system improves often accuracy of diagnostic information compared to conventional X-ray examinations. Pharmacoradiography should be used more often; the examiner has to be familiar with the principal effects and side effects of the pharmaceutica. The present paper gives a short summary about drugs and examination techniques most often used.


Influence of Disease Duration on Circulating Levels of miRNAs in Children and Adolescents with New Onset Type 1 Diabetes.
Circulating microRNAs (miRNAs) have been implicated in several pathologies including type 1 diabetes. In the present study, we aimed to identify circulating miRNAs affected by disease duration in children with recent onset type 1 diabetes. Forty children and adolescents from the Danish Remission Phase Cohort were followed with blood samples drawn at 1, 3, 6, 12, and 60 months after diagnosis. Pancreatic autoantibodies were measured at each visit. Cytokines were measured only the first year. miRNA expression profiling was performed by RT-qPCR. The effect of disease duration was analyzed by mixed models for repeated measurements adjusted for sex and age. Eight miRNAs (hsa-miR-10b-5p, hsa-miR-17-5p, hsa-miR-30e-5p, hsa-miR-93-5p, hsa-miR-99a-5p, hsa-miR-125b-5p, hsa-miR-423-3p, and hsa-miR-497-5p) were found to significantly change in expression (adjusted p-value < 0.05) with disease progression. Three pancreatic autoantibodies, ICA, IA-2A, and GAD65A, and four cytokines, IL-4, IL-10, IL-21, and IL-22, were associated with the miRNAs at different time points. Pathway analysis revealed associations with various immune-mediated signaling pathways. Eight miRNAs that were involved in immunological pathways changed expression levels during the first five years after diagnosis and were associated with variations in cytokine and pancreatic antibodies, suggesting a possible effect on the immunological processes in the early phase of the disease.


Facing the Growing COVID-19 Infodemic: Digital Health Literacy and Information-Seeking Behaviour of University Students in Slovenia.
The entire world is faced with the COVID-19 pandemic, which is also accompanied by an infodemic. This refers to the rapid spread of (accurate and false) information, mainly through internet usage increasing. Digital health literacy (DHL) is therefore important for addressing challenges related to online health information and services, as well as for navigation through the complex information landscape with huge amounts of different (and conflicting) information about COVID-19. The aim of this study is to examine the level of DHL in relation to COVID-19 in Slovenian university students and to determine online information-seeking behaviour in order to plan and prepare effective communication interventions for this sub-population. A cross-sectional survey, administered by an online questionnaire, was conducted to collect data on DHL. A total of 3621 students participated, of whom 70% were female and the average age was 22.65 years (SD = 4.65). Bivariate analyses were performed to assess the association of key characteristics with DHL. Overall, the results show that the level of DHL among students is sufficient. Most difficulties were reported in assessing the reliability of information (n = 1484, 49.3%). Approximately one third of the students (n = 847, 27.9%) reported having problems in finding information of their interest, and somewhat more (n = 900, 29.6%) reported difficulties in making a selection among all the information found. Students with a sufficient level of DHL are more likely to seek information through search engines and websites of official institutions, while students with a limited level of DHL more frequently reported using social media for health information searches. It is necessary to establish interventions for a systematic lift of the DHL and health literacy (HL) of all population groups.


Effects of silica on lung collagen.
A single intratracheal injection of 50 mg crystalline silica (quartz) into rats produces silicosis. This animal model may be used to study collagen metabolism during the early, middle, and late phases of lung injury, corresponding respectively to the stages of lung injury, development of discrete granulomas, and development of mature silicotic nodules. The early phase is characterized by a rapid increase in the rate of synthesis of lung collagen (within one week of instillation) and increased deposition of excess lung collagen (significant increases within two weeks of instillation). Later phases are characterized by a continuing increase in deposition of excess lung collagen for at least one year after instillation. Silica-induced fibrosis is unique among all the animal models (and in most human fibrotic diseases) thus far examined, in that the excess collagen deposited in the lung contains normal ratios of the two major collagen types of the lung: types I and III. This collagen is nonetheless biochemically different from normal lung collagen. There are reproducible and characteristic differences in the intermolecular cross-links of the collagen in lungs from rats injected with silica. Within one month of silica instillation (the earliest time point examined thus far), an increased hydroxylysine content of collagen can be appreciated. The reducible dysfunctional cross-links are also more likely to be derived from hydroxylysine (i.e. the ratio of dihydroxylated to monohydroxylated cross-links increases). Within four months of silica instillation (and increasingly thereafter), increased amounts of the mature trifunctional cross-link hydroxypyridinium (derived from three residues of hydroxylysine) can also be appreciated, seemingly paralleling the evolution of mature silicotic nodules in these lungs. These changes in cross-linking of lung collagen seem to be common to all the animal models of pulmonary fibrosis examined, and are also consistent with changes occurring in human fibrotic lungs. Preliminary observations suggest that the locus of cross-linking remains the same: hydroxylysine replaces lysine in the primary structure of a specific collagen alpha chain to form the altered cross-links. Thus, there may be molecular markers for the collagen of fibrosis in diseased lungs.


Quality control in the apoA-I secretory pathway: deletion of apoA-I helix 6 leads to the formation of cytosolic phospholipid inclusions.
From a total of 47 known apolipoprotein A-I (apoA-I) mutations, only 18 are linked to low plasma HDL apoA-I concentrations, and 78% of these map to apoA-I helices 6 and 7 (residues 143-186). Gene transfer and transgenic mouse studies have shown that several helix 6 apoA-I mutations have reduced hepatic HDL production. Our objective was to examine the impact of helix 6 modifications on intracellular biosynthetic processing and secretion of apoA-I. Cells were transfected with wild-type or mutant apoA-I, radiolabeled with [(35)S]Met/Cys, and then placed in unlabeled medium for up to 4 h. Results show that >90% of newly synthesized wild-type apoA-I was secreted by 60 min. Over the same length of time, only 20% of helix 6 deletion mutant (Delta 6 apoA-I) was secreted, whereas 80% remained cell associated. Microscopic and biochemical studies revealed that cell-associated Delta 6 apoA-I was located predominantly within the cytoplasm as lipid-protein inclusions, whereas wild-type apoA-I was localized in the endoplasmic reticulum/Golgi. Results using other helix deletions or helix 6 substitution mutations indicated that only complete removal of helix 6 resulted in massive cytoplasmic accumulation. These data suggest that alterations in native apoA-I conformation can lead to aberrant trafficking and accumulation of apolipoprotein-phospholipid structures. Thus, conformation-dependent alterations in intracellular trafficking and turnover may underlie the reduced plasma HDL concentrations observed in individuals harboring deletion mutations within helix 6.


miR-335 and miR-363 regulation of neuroblastoma tumorigenesis and metastasis.
microRNA (miRNA) functions broadly as post-transcriptional regulators of gene expression, and disproportionate miRNAs can result in dysregulation of oncogenes in cancer cells. We have previously shown that gastrin-releasing peptide receptor (GRP-R) signaling regulates tumorigenicity of neuroblastoma cells. Herein, we sought to characterize miRNA profile in GRP-R silenced neuroblastoma cells, and to determine the role of miRNAs on tumorigenicity and metastatic potential. Human neuroblastoma cell lines, BE(2)-C and SK-N-SH, were used for our study. Stably transfected GRP-R silenced cells were assessed for miRNA profiles. Cells were transfected with miR-335, miR-363, or miR-CON, a nontargeting control, and in vitro assays were performed. In vivo functions of miR-335 and miR-363 were also assessed in a spleen-liver metastasis murine model. GRP-R silencing significantly increased expression of miR-335 and miR-363 in BE(2)-C cells. Overexpression of miR-335 and miR-363 decreased tumorigenicity as measured by clonogenicity, anchorage-independent growth, and metastasis determined by cell invasion assay and liver metastasis in vivo. We report, for the first time, that GRP-R-mediated tumorigenicity and increased metastatic potential in neuroblastoma are regulated, in part, by miR-335 and miR-363. A better understanding of the anti-tumor functions of miRNAs could provide valuable insights to discerning molecular mechanisms responsible for neuroblastoma metastasis.


Neuropsychological performance in patients with depression is associated with clinical, etiological and genetic risk factors.
The present study explores neuropsychological functioning in patients with depression with reference to key clinical, etiological and genetic features. In comparison to healthy volunteers, patients with severe depression demonstrated poorer performance on all neuropsychological tests except for WAIS-R Vocabulary and a 64-item computerized version of the Wisconsin Card Sorting Test. On average, patients exhibited significant impairments (greater than -2 standard deviation units) on tests of simple reaction time, Part B of the Trail Making Test and Raven's Colored Progressive Matrices. When performance decrements were analyzed with reference to key clinical features, patients with melancholia performed more poorly on WAIS-R Vocabulary, semantic fluency and choice reaction time than those with nonmelancholic depression. After controlling for age, those patients with late-onset depression (i.e., age of onset > or =50 years) exhibited poorer performance on a computerized version of the Tower of London test in comparison to those with an early onset. While there was no relationship between neuropsychological test scores and summed vascular risk factors or apolipoprotein E genotypes, presence of the methylenetetrahydrofolate reductase gene mutation was associated with slowed reaction time. The differential relationships between clinical, etiological and genetic risks and neuropsychological performance supports the presence of unique pathophysiological mechanisms in distinct subgroups of patients. These findings underscore the need to consider subtypes when investigating neuropsychological deficits in patients with depression.


Representation of Hispanics in the National Marrow Donor Program.
Since less than one-third of patients in need of a BMT find related donors, most patients will rely on registries of volunteer donors. For patients from minority ethnic groups the chances of finding matched unrelated donors are lower, in part due to the smaller representation of minorities in the registries. Our purpose was to determine the representation of Hispanics in the National Marrow Donor Program (NMDP), the largest registry of volunteer marrow donors in the United States. We analyzed a database provided by the NMDP that contained information on minorities. The number of Hispanic volunteer donors has increased 110-fold in the last 6 years. The proportion of Hispanics in the registry has also increased from 1.1% to 6%. Nevertheless, the proportion of Hispanic patients that received unrelated marrow transplants facilitated by the NMDP has increased only from 2.8% to 3.9% since 1989. Only 19.7% of the formal searches initiated by Hispanic patients resulted in transplants compared to the 30.4% observed in the Caucasian population. Despite increments in the number and proportion of Hispanic volunteer donors, the proportion of Hispanics that receive BMT from unrelated donors remains low. We conclude that, in addition to increased recruitment efforts, other strategies will be necessary in order to find enough marrow donors to meet the needs of the Hispanic population.


Ribosome Rescue Inhibitors Kill Actively Growing and Nonreplicating Persister Mycobacterium tuberculosis Cells.
The emergence of Mycobacterium tuberculosis (MTB) strains that are resistant to most or all available antibiotics has created a severe problem for treating tuberculosis and has spurred a quest for new antibiotic targets. Here, we demonstrate that trans-translation is essential for growth of MTB and is a viable target for development of antituberculosis drugs. We also show that an inhibitor of trans-translation, KKL-35, is bactericidal against MTB under both aerobic and anoxic conditions. Biochemical experiments show that this compound targets helix 89 of the 23S rRNA. In silico molecular docking predicts a binding pocket for KKL-35 adjacent to the peptidyl-transfer center in a region not targeted by conventional antibiotics. Computational solvent mapping suggests that this pocket is a druggable hot spot for small molecule binding. Collectively, our findings reveal a new target for antituberculosis drug development and provide critical insight on the mechanism of antibacterial action for KKL-35 and related 1,3,4-oxadiazole benzamides.


The effect of hypothermia therapy on cortical laminar disruption following ischemic injury in neonatal mice.
Hypothermia has been proposed as a treatment for reducing neuronal damage in the brain induced by hypoxic ischemia. In the developing brain, hypoxic ischemia-induced injury may give rise to cerebral palsy (CP). However, it is unknown whether hypothermia might affect the development of CP. The purpose of this study was to investigate whether hypothermia would have a protective effect on the brains of immature, 3-day old (P3) mice after a challenge of cerebral ischemia. Cerebral ischemia was induced in P3 mice with a right common carotid artery ligation followed by hypoxia (6% O2, 37°C) for 30 min. Immediately after hypoxic ischemia, mice were exposed to hypothermia (32°C) or normothermia (37°C) for 24 h. At 4 weeks of age, mouse motor development was tested in a behavioral test. Mice were sacrificed at P4, P7, and 5 weeks to examine brain morphology. The laminar structure of the cortex was examined with immunohistochemistry (Cux1/Ctip2); the number of neurons was counted; and the expression of myelin basic protein (MBP) was determined. The hypothermia treatment was associated with improved neurological outcomes in the behavioral test. In the normothermia group, histological analyses indicated reduced numbers of neurons, reduced cortical laminar thickness in the deep, ischemic cortical layers, and significant reduction in MBP expression in the ischemic cortex compared to the contralateral cortex. In the hypothermia group, no reductions were noted in deep cortical layer thickness and in MBP expression in the ischemic cortex compared to the contralateral cortex. At 24 h after the hypothermia treatment prevented the neuronal cell death that had predominantly occurred in the ischemic cortical deep layers with normothermia treatment. Our findings may provide a preclinical basis for testing hypothermal therapies in patients with CP induced by hypoxic ischemia in the preterm period.


New Continuum Approaches for Determining Protein-Induced Membrane Deformations.
The influence of the membrane on transmembrane proteins is central to a number of biological phenomena, notably the gating of stretch activated ion channels. Conversely, membrane proteins can influence the bilayer, leading to the stabilization of particular membrane shapes, topological changes that occur during vesicle fission and fusion, and shape-dependent protein aggregation. Continuum elastic models of the membrane have been widely used to study protein-membrane interactions. These mathematical approaches produce physically interpretable membrane shapes, energy estimates for the cost of deformation, and a snapshot of the equilibrium configuration. Moreover, elastic models are much less computationally demanding than fully atomistic and coarse-grained simulation methodologies; however, it has been argued that continuum models cannot reproduce the distortions observed in fully atomistic molecular dynamics simulations. We suggest that this failure can be overcome by using chemically and geometrically accurate representations of the protein. Here, we present a fast and reliable hybrid continuum-atomistic model that couples the protein to the membrane. We show that the model is in excellent agreement with fully atomistic simulations of the ion channel gramicidin embedded in a POPC membrane. Our continuum calculations not only reproduce the membrane distortions produced by the channel but also accurately determine the channel's orientation. Finally, we use our method to investigate the role of membrane bending around the charged voltage sensors of the transient receptor potential cation channel TRPV1. We find that membrane deformation significantly stabilizes the energy of insertion of TRPV1 by exposing charged residues on the S4 segment to solution.


Development of a completely encapsulated intraocular pressure sensor.
A completely encapsulated intraocular pressure (IOP) sensor equipped with telemetric signal and energy transfer is introduced integrated into a silicone disc for implantation into the eye. After implantation into enucleated pig eyes and into rabbit eyes in vivo, the IOP was recorded and compared to established techniques of IOP measurement. Pressure chamber tests showed that the sensor functioned correctly after biocompatible encapsulation in polydimethylsiloxane. In vivo and in vitro tests in rabbit and pig eyes demonstrated that the implanted system worked with the same precision as established techniques for IOP determination. The correlation between the measurements with the implanted device and pneumotonometry in several experiments was between 0.9 and 0.99. This device serves as a functioning model for the realization of a telemetric IOP sensor for integration into an artificial intraocular lens. Such a device will open new perspectives, not only in the management of glaucoma, but also in basic research for mechanisms of glaucoma.


The role of venous ultrasonography in the diagnosis of suspected deep venous thrombosis and pulmonary embolism.
This paper describes the role of venous ultrasonography in the diagnosis of suspected deep venous thrombosis and pulmonary embolism. Inability to compress the common femoral or popliteal vein is usually diagnostic of a first episode of deep venous thrombosis in symptomatic patients (positive predictive value of about 97%). Full compressibility of both of these sites excludes proximal deep venous thrombosis in symptomatic patients (negative predictive value of about 98%). In patients with suspected deep venous thrombosis or in those who present with suspected pulmonary embolism but have a nondiagnostic lung scan, the subsequent risk for symptomatic venous thromboembolism is very low (<2% during 6 months of follow-up) provided that ultrasonography of the proximal veins remains normal in the course of 1 week (suspected deep venous thrombosis) or 2 weeks (suspected pulmonary embolism). Anticoagulation and further diagnostic testing can usually be safely withheld in these situations. Venous ultrasonography is much less reliable for the diagnosis of asymptomatic, isolated distal, and recurrent deep venous thrombosis than for the diagnosis of a first episode of proximal deep venous thrombosis in symptomatic patients. Clinical evaluation of the probability of deep venous thrombosis or pulmonary embolism, preferably by using a validated clinical model, complements venous ultrasonographic findings and helps to identify patients who would benefit from additional (often invasive) diagnostic testing. Thus, venous ultrasonography is thought to be a very valuable test for the diagnosis and management of patients with suspected deep venous thrombosis or pulmonary embolism.


Plasma selenium is positively related to performance in neurological tasks assessing coordination and motor speed.
Parkinson's disease (PD) is a degenerative process affecting the striato nigral system (SN). Its etiology, although obscure, may involve oxidative damage. Selenium, an antioxidant, was shown to protect the SN in animal models. In the current study, we investigate the association between plasma selenium concentrations and the presence of "soft" neurological signs related to the SN. Plasma selenium concentration was assessed in participants of age ≥65 years in the InCHIANTI study, a population-based cohort study in Tuscany, Italy. PD was defined based on standard criteria. "Soft" neurological signs were ascertained by physical examination. A total of 1,012 participants were included. No association was found between the presence of PD and plasma selenium. There was, however, a strong association between plasma selenium and timed performance-based assessments. Lower levels of selenium were significantly associated withdecreased performance in neurological tests of coordination among older adults. Prospective studies are needed to further investigate the effects of selenium on SN dysfunction.


The human function curve - a paradigm for our times.
Persistent overloading causes fatigue followed by exhaustion and then ill-health resulting from homeostasis violation and reduced resistance. A breakdown is inevitable if the struggle continues. This course can be illustrated by a shaped performance-arousal curve. The paradigm clarifies the effects of psychosocial handicaps (low curves), of training (high curves), of arousal reduction (= relaxation), and of arousal rising from normal to destructive levels. The paradigm accommodates catastrophe theory and provides an unambiguous basis for screening and intervention in hypertension and coronary heart disease where there is a self-defeating struggle to close the ever widening gap between the actual performance capability and the intended.


Genetic differences in cytochrome P450 enzymes and antidepressant treatment response.
Antidepressant response varies between patients, possibly due to differences in the rate cytochrome P450 enzymes metabolise antidepressants into inactive compounds. Drug metabolism rates are influenced by common variants in the genes encoding these enzymes. However, it remains unclear whether treatment outcomes can be predicted by either CYP450 genotype or antidepressant serum concentration. In GENDEP (a pharmacogenetic study of depressed individuals treated with either escitalopram or nortriptyline), serum concentrations of antidepressants and their primary metabolite were measured after eight weeks treatment and variants in CYP2D6 and CYP2C19 were genotyped. Amongst patients taking escitalopram (n=223), the genotype CYP2C19 was significantly associated with escitalopram serum concentrations and desmethylescitalopram:escitalopram ratio. For those taking nortriptyline (n=161), the CYP2D6 genotype was significantly associated with nortriptyline and 10-hydroxynortriptyline serum concentrations and 10-hydroxynortriptyline:nortrip-tyline ratio. CYP450 genotypes conferring greater enzyme activity were linked to lower drug serum concentrations and higher metabolite:drug ratios. Nonetheless, no significant association was found between either CYP450 genotype or antidepressant serum concentration and treatment response. While there is a significant relationship between the CYP450 genotype and serum concentrations of escitalopram and nortriptyline, the genotypes are not predictive of differences in treatment response for either drug. Furthermore, differences in antidepressant serum concentrations are not associated with variability in treatment response.