The zebrafish secretome.
The secretome is a functionally rich proteome subset, including cellular membrane and extracellular proteins processed through the secretory pathway. In this study, Danio rerio and Homo sapiens RefSeq proteins were analyzed with SignalP, TargetP, Phobius, and pTarget algorithms. About 16.5% of the zebrafish proteome and 17.0% of the human proteome possessed predicted N-terminal signal sequences. Nearly half of these proteins were subsequently classified as soluble, as they lacked predicted transmembrane domains. The soluble proteins were further subclassified, predicting 1345 (3.8%) zebrafish and 1207 (3.2%) human proteins as extracellular. Comparison of the zebrafish and human soluble secretome proteins identified 372 as orthologs, on the basis of reciprocal BLAST best hits. The computational characterization of the zebrafish proteins found many more members of the secretome than annotated in the SwissProt database. Only 180 of the 2078 zebrafish SwissProt protein entries, and 995 of the 19,294 human SwissProt protein entries were annotated with secreted protein locales. A specific investigation of the fibroblast growth factor and matrix metalloproteinase (MMP) protein families confirmed the prediction data and generated annotation of three additional putative MMP zebrafish proteins. This study presents the first known published description of the zebrafish secretome since the completion of the zebrafish genome sequencing project.


Observation of Scaling in the Dynamics of a Strongly Quenched Quantum Gas.
We report on the experimental observation of scaling in the time evolution following a sudden quench into the vicinity of a quantum critical point. The experimental system, a two-component Bose gas with coherent exchange between the constituents, allows for the necessary high level of control of parameters as well as the access to time-resolved spatial correlation functions. The theoretical analysis reveals that when quenching the system close to the critical point, the energy introduced by the quench leads to a short-time evolution exhibiting crossover reminiscent of the finite-temperature critical properties in the system's universality class. Observing the time evolution after a quench represents a paradigm shift in accessing and probing experimentally universal properties close to a quantum critical point and allows in a new way benchmarking of quantum many-body theory with experiments.


Foamy virus nuclear RNA export is distinct from that of other retroviruses.
Most retroviruses express all of their genes from a single primary transcript. In order to allow expression of more than one gene from this RNA, differential splicing is extensively used. Cellular quality control mechanisms retain and degrade unspliced or partially spliced RNAs in the nucleus. Two pathways have been described that explain how retroviruses circumvent this nuclear export inhibition. One involves a constitutive transport element in the viral RNA that interacts with the cellular mRNA transporter proteins NXF1 and NXT1 to facilitate nuclear export. The other pathway relies on the recognition of a viral RNA element by a virus-encoded protein that interacts with the karyopherin CRM1. In this report, we analyze the protein factors required for the nuclear export of unspliced foamy virus (FV) mRNA. We show that this export is CRM1 dependent. In contrast to other complex retroviruses, FVs do not encode an export-mediating protein. Cross-linking experiments indicated that the cellular protein HuR binds to the FV RNA. Inhibition studies showed that both ANP32A and ANP32B, which are known to bridge HuR and CRM1, are essential for FV RNA export. By using this export pathway, FVs solve a central problem of viral replication.


Challenges of implementing point-of-care testing (POCT) glucose meters in a pediatric acute care setting.
To investigate factors contributing to analytical bias in POCT glucose values generated by the NICU versus the core laboratory. The LifeScan Flexx hospital system glucose meters (SureStep) were used in precision and comparison studies between the NICU and laboratory (ABL715 and Vitros 950). Analysis of 40 neonatal blood samples revealed a positive bias between the NICU glucose meters versus either the laboratory glucose meter or instrument (mean difference of 0.28 and 0.21 mmol/L, respectively). Linear regression analysis (R2 = 0.0584) of the difference in glucose results versus time elapsed between measurements indicated that the bias observed between the NICU and laboratory glucose meters was not due to in vitro glycolysis for samples transported on ice. Further analysis indicated that the bias appeared to be mostly operator driven, with different NICU operators exhibiting different mean biases. Increasing the amount of blood applied to the SureStep Pro test strip (e.g., 60 vs. 20 microL), led to higher values for glucose concentration for the same blood. Nearly 50% of all glucose values reported for the NICU were obtained by the SureStep Flexx glucose meters in a 3-month period following the introduction of POCT, yet the number of laboratory-reported glucose results for the same period increased by 21% as compared to the previous year. Operator error appears to be a source of bias present between the NICU and laboratory, and despite glucose meter utilization in the NICU, the number of glucose measurements by the central laboratory increased after POCT introduction.


[A 30-year experience with combined radiotherapy of cervix cancer].
The development of the method of complex radiation treatment at the Institute in 1946-1980 is followed. Clinical data on 2,313 patients with stage I, II and III carcinoma of the uterine cervix formed the basis of a multifactor analysis. Five-year survival rates and frequency of complications following different modalities of radiation therapy were studied. The 5-year survival rate in cases of orthovolt X-ray therapy or intracavitary gammatherapy from unfixed sources was 54.2% (all stages of carcinoma). The results were improved by 11.9% (66.3% versus 54.2%) when remote gammatherapy using increased focal doses in the zones of parametral and lymphogenic metastatic spread was employed and the exposed volume and the focus of lesion were properly aligned. The development of pretreatment X-ray topometric procedures was followed by a further improvement in the results (72.2% as compared with 60.7%), particularly in cases of stage III carcinoma. The 5-year survival rate rose to 80.1% (all stages) as a result of application of complex radiation therapy preceded by suitable topometric preparation.


p38 MAP kinase is necessary for melanoma-mediated regulation of VE-cadherin disassembly.
Vascular endothelial (VE)-cadherin is localized to the endothelial borders and the adherens junctions, which are regulated by changes in mitogen-activated protein (MAP) kinases, GTPases, and intracellular calcium. We previously showed that melanoma cells induce VE-cadherin disassembly through contact with human umbilical vein endothelial cells in coculture. However, the exact mechanism by which melanoma cells signal endothelial cells to induce VE-cadherin junction disassembly is not well understood. In this study, VE-cadherin junction disassembly was further examined under fluorescence microscopy. We found that melanoma-induced VE-cadherin junction disassembly and upregulation of p38 MAP kinase in endothelial cells is regulated by both soluble factors from melanomas, particularly interleukin (IL)-8, IL-6, and IL-1beta, and through vascular cell adhesion molecule-1. Neutralizing melanoma-secreted soluble factors reduced endothelial gap formation. Endothelial cells transfected with MAP kinase kinase 6, a direct activator of p38 MAP kinase, increased VE-cadherin-mediated gap formation, facilitating melanoma transendothelial migration. In contrast, endothelial cells transfected with small-interfering RNA against p38 MAP kinase expression largely prevented melanoma transendothelial migration in Boyden chamber experiments. These findings indicate that p38 MAP kinase proteins regulate VE-cadherin junction disassembly, facilitating melanoma migration across endothelial cells.


Ferroelectric polymer scaffolds based on a copolymer of tetrafluoroethylene with vinylidene fluoride: fabrication and properties.
A solution blow spinning technique is a method developed recently for making nonwoven webs of micro- and nanofibres. The principal advantage of this method compared to a more traditional electrospinning process is its significantly higher production rate. In this work, the solution blow spinning method was further developed to produce nonwoven polymeric scaffolds based on a copolymer of tetrafluoroethylene with vinylidene fluoride solution in acetone. A crucial feature of the proposed method is that high-voltage equipment is not required, which further improves the method's economics. Scanning electron microscopy analysis of the samples demonstrated that the surface morphology of the nonwoven materials is dependent on the polymer concentration in the spinning solution. It was concluded that an optimum morphology of the nonwoven scaffolds for medical applications is achieved by using a 5% solution of the copolymer. It was established that the scaffolds produced from the 5% solution have a fractal structure and anisotropic mechanical properties. X-ray diffraction, infrared spectroscopy, Raman spectroscopy and differential scanning calorimetry demonstrated that the fabricated nonwoven materials have crystal structures that exhibit ferroelectric properties. Gas chromatography has shown that the amount of acetone in the nonwoven material does not exceed the maximum allowable concentration of 0.5%. In vitro analysis, using the culture of motile cells, confirmed that the nonwoven material is non-toxic and does not alter the morpho-functional status of stem cells for short-term cultivation, and therefore can potentially be used in medical applications.


Clinician-targeted intervention and patient-reported counseling on physical activity.
Limited time and lack of knowledge are barriers to physical activity counseling in primary care. The objective of this study was to examine the effectiveness of a clinician-targeted intervention that used the 5As (Ask, Advise, Agree, Assist, Arrange) approach to physical activity counseling in a medically underserved patient population. Family medicine clinicians at 2 community health centers were randomized to Group 1 or Group 2 intervention. Both clinician groups participated in 4 training sessions on the 5As for physical activity counseling; Group 2 training took place 8 months after Group 1 training. Both groups were trained to refer patients to a community exercise program. We used a pre-post analysis to evaluate the effectiveness of the intervention on clinician use of 5As. Eligible patients (n = 319) rated their clinicians' counseling skills by using a modified Physical Activity Exit Interview (PAEI) survey. Clinicians (n = 10) self-assessed their use of the 5As through a survey and interviews. Both patient and clinician groups had similar sociodemographic characteristics. The PAEI score for both groups combined increased from 6.9 to 8.6 (on a scale of 0-15) from baseline to immediately postintervention (P = .01) and was 8.2 (P = .09) at 6-month follow-up; most of the improvement in PAEI score was due to increased use of 5As skills by Group 2 clinicians. Group 1 reported difficulty with problem solving, whereas Group 2 reported ease of referral to the community exercise program. A clinician training intervention showed mixed results for 5As physical activity counseling.


Performance of the ATLAS trigger system in 2015.
During 2015 the ATLAS experiment recorded [Formula: see text] of proton-proton collision data at a centre-of-mass energy of [Formula: see text]. The ATLAS trigger system is a crucial component of the experiment, responsible for selecting events of interest at a recording rate of approximately 1 kHz from up to 40 MHz of collisions. This paper presents a short overview of the changes to the trigger and data acquisition systems during the first long shutdown of the LHC and shows the performance of the trigger system and its components based on the 2015 proton-proton collision data.


Dynamical state transitions into addictive behaviour and their early-warning signals.
The theory of critical transitions in complex systems (ecosystems, climate, etc.), and especially its ability to predict abrupt changes by early-warning signals based on analysis of fluctuations close to tipping points, is seen as a promising avenue to study disease dynamics. However, the biomedical field still lacks a clear demonstration of this concept. Here, we used a well-established animal model in which initial alcohol exposure followed by deprivation and subsequent reintroduction of alcohol induces excessive alcohol drinking as an example of disease onset. Intensive longitudinal data (ILD) of rat drinking behaviour and locomotor activity were acquired by a fully automated drinkometer device over 14 weeks. Dynamical characteristics of ILD were extracted using a multi-scale computational approach. Our analysis shows a transition into addictive behaviour preceded by early-warning signals such as instability of drinking patterns and locomotor circadian rhythms, and a resultant increase in low frequency, ultradian rhythms during the first week of deprivation. We find evidence that during prolonged deprivation, a critical transition takes place pushing the system to excessive alcohol consumption. This study provides an adaptable framework for processing ILD from clinical studies and for examining disease dynamics and early-warning signals in the biomedical field.


Efficacy of electrical stimulation to increase muscle strength in people with neurological conditions: a systematic review.
Weakness in partially paralysed muscles is a disabling impairment for people with neurological conditions. Strength training programmes are widely administered to address this impairment. There is a common belief that the effectiveness of strength training programmes can be enhanced by the addition of electrical stimulation. The purpose of this systematic review was to assess the efficacy of electrical stimulation for increasing voluntary strength in people with neurological conditions. Eligible randomized trials of electrical stimulation were identified by searches of computerized databases. The search yielded 11,267 abstracts, of which 60 were retrieved. Two assessors independently reviewed full text versions of these articles. Eighteen studies satisfied the inclusion criteria. These studies involved participants with spina bifida (n = 1), cerebral palsy (n = 1), peripheral nerve lesion (n = 1), multiple sclerosis (n = 1), spinal cord injury (n = 3) and stroke (n = 11). The mean (SD) PEDro score for trial quality was 4.9 (1.0) out of 10. Meta-analyses of studies involving similar patients were not done because of insufficient data or lack of homogeneity. The results of all studies were analysed individually. Several studies suggest a modest beneficial effect of electrical stimulation in patients with stroke. It is not clear whether patients with other types of neurological disabilities benefit from electrical stimulation in the same way.


Mutational analysis of PVX TGBp3 links subcellular accumulation and protein turnover.
Potato virus X (PVX) TGBp3 is required for virus cell-to-cell transport, has an N-terminal transmembrane domain, and a C-terminal cytosolic domain. In the absence of virus infection TGBp3:GFP is seen in the cortical and perinuclear ER. In PVX infected cells the TGBp3:GFP fusion is also seen in the nucleoplasm indicating that events during PVX infection trigger entry into the nucleus. Mutational analysis failed to identify a nuclear targeting domain. Mutations inhibiting TGBp3 association with the ER and inhibiting virus movement did not block TGBp3:GFP in the nucleoplasm. A mutation disrupting the N-terminal transmembrane domain of TGBp3 caused the fusion to accumulate in the nucleus indicating that nuclear import is regulated by ER interactions. Tunicamycin, an ER-stress inducing chemical, caused lower levels of GFP and TGBp3:GFP to accumulate in virus infected protoplasts. MG115 and MG132 were used to demonstrate that wild-type and mutant TGBp3:GFP fusions were degraded by the 26S proteasome. These observations are consistent with an ER-associated protein degradation (ERAD) pathway suggesting that PVX TGBp3, similar to aberrant ER proteins, is translocated to the cytoplasm for degradation. Nuclear accumulation of mutant and wild-type TGBp3:GFP is independent of other PVX proteins and may be another feature of an ERAD pathway.


Phyto-oestrogens and cancer.
Phyto-oestrogens have been suggested to have a preventive effect against various cancers. This review includes a discussion of the consumption of phyto-oestrogen-rich foods such as soy, a source of isoflavones, and whole grain products, which contain lignans, and their role in the prevention of breast, prostate, and colon cancer. In women, a soy-containing diet is only slightly protective against breast cancer, if at all, but is more likely to be beneficial if initiated before puberty or during adolescence. These findings are supported by conclusions of studies of immigrants and other epidemiological studies. However, in one case-control study and one prospective study, a low-lignan diet increased the risk of breast cancer. Experimental evidence also exists for an inhibitory effect of soy and rye bran on prostate-cancer growth and for rye bran or isolated lignans on colon cancer. Whether these observed protective effects are caused by the presence of dietary phyto-oestrogens, or whether they are merely indicators of a healthy diet in general, has not been established.


The Effect of Interfacial Chemical Bonding in TiO2-SiO2 Composites on Their Photocatalytic NOx Abatement Performance.
The chemical bonding of particulate photocatalysts to supporting material surfaces is of great importance in engineering more efficient and practical photocatalytic structures. However, the influence of such chemical bonding on the optical and surface properties of the photocatalyst and thus its photocatalytic activity/reaction selectivity behavior has not been systematically studied. In this investigation, TiO2 has been supported on the surface of SiO2 by means of two different methods: (i) by the in situ formation of TiO2 in the presence of sand quartz via a sol-gel method employing tetrabutyl orthotitanium (TBOT); and (ii) by binding the commercial TiO2 powder to quartz on a surface silica gel layer formed from the reaction of quartz with tetraethylorthosilicate (TEOS). For comparison, TiO2 nanoparticles were also deposited on the surfaces of a more reactive SiO2 prepared by a hydrolysis-controlled sol-gel technique as well as through a sol-gel route from TiO2 and SiO2 precursors. The combination of TiO2 and SiO2, through interfacial Ti-O-Si bonds, was confirmed by FTIR spectroscopy and the photocatalytic activities of the obtained composites were tested for photocatalytic degradation of NO according to the ISO standard method (ISO 22197-1). The electron microscope images of the obtained materials showed that variable photocatalyst coverage of the support surface can successfully be achieved but the photocatalytic activity towards NO removal was found to be affected by the preparation method and the nitrate selectivity is adversely affected by Ti-O-Si bonding.


Partners or partisans? Patient participation at Marylebone Health Centre.
This paper outlines some of the issues which arose for patients and professionals involved in patient participation projects at the Marylebone health centre in London. It describes the projects undertaken and focuses on the practical implications of working with rather than for patients. Dilemmas surrounding patient participation are discussed, including the ways volunteers are rewarded, how doctors and patients can share knowledge, how participation is affected by professional boundaries, and why a regular group meeting may not necessarily be the best way to involve patients in decision making. The successes of patient participation are also highlighted.


Low-dose-rate vs. high-dose-rate intracavitary brachytherapy for carcinoma of the cervix: The University of Alabama at Birmingham (UAB) experience.
To review the clinical outcome retrospectively of cervical cancer patients treated definitively with either high-dose-rate (HDR) or low-dose-rate (LDR) brachytherapy. One hundred sixty patients (44 Stage I, 83 Stage II, and 33 Stage III) were treated from 1990 to 2000 with curative intent for carcinoma of the cervix. One hundred three LDR patients were compared to 57 HDR patients. Two groups were treated during the same period. An external beam dose of 45 Gy to the entire pelvis was delivered at 1.8 Gy per fraction to most patients before the first intracavitary insertion in both groups. Brachytherapy was delivered in one to two LDR implants or four to five HDR implants at 6 Gy per fraction. The prescribed dose to Point A for LDR was at least 80-85 Gy. Patient characteristics were similar for each cohort. Point A doses were similar for each stage. The primary endpoints assessed were survivals and failure sites. Endpoints were estimated using the Kaplan-Meier method and comparisons between treatment groups were performed using the log-rank test. The median followup was 48 months for the LDR group and 59 months for the HDR group. For all stages combined and stage for stage in both groups, there was no statistically significant difference in locoregional control, cause-specific survival, and overall survival for LDR compared with HDR. Locoregional control and overall survival were 78% and 60% for LDR compared to 76% and 55% for HDR at 3 years, respectively (p = 0.96 and p = 0.48). Median cause-specific survival values for LDR vs. HDR were 71 and 81 months, respectively (p = 0.62). The cause-specific survival for LDR patients was 62% compared with 59% for HDR patients at 3 years. For Stage IB2, II, and III LDR patients, cause-specific survival rates were 62%, 67%, and 45%, compared to 67%, 57%, and 33% for HDR at 3 years, respectively (p = 0.75, p = 0.95, and p = 0.48). For patients with a recorded site of first failure, the most common site was locoregional (56%) and then distant metastases (26%). Eight patients who were cancer free developed late complications requiring surgical intervention. Two patients were in the HDR group (3.5%) and 5 in the LDR group (4.8%). Similar outcome was observed for LDR compared with HDR intracavitary brachytherapy for the entire cohort. In this review, HDR group was not inferior to LDR group in advanced stages. This is likely because our patients were treated with brachytherapy after a high dose of external pelvic radiotherapy in both LDR and HDR patients.


The SCHOOL of nature: II. Protein order, disorder and oligomericity in transmembrane signaling.
Recent reports have revealed that many proteins that do not adopt globular structures under native conditions, thus termed intrinsically disordered proteins (IDPs), are involved in cell signaling. Intriguingly, physiologically relevant oligomerization of IDPs has been recently observed and shown to exhibit unique biophysical characteristics, including the lack of significant changes in chemical shift and peak intensity upon binding. In this work, I summarize several distinct features of protein disorder that are especially important as related to receptor-mediated transmembrane signal transduction. I also hypothesize that interactions of IDPs with their protein or lipid partners represent a general biphasic process with the "no disorder-to-order" fast interaction which, depending on the interacting partner, may or may not be accompanied by the slow formation of a secondary structure. Further, I suggest signaling-related functional connections between protein order, disorder, and oligomericity and hypothesize that receptor oligomerization induced or tuned upon ligand binding outside the cell is translated across the membrane into protein oligomerization inside the cell, thus providing a general platform, the Signaling Chain HOmoOLigomerization (SCHOOL) platform, for receptor-mediated signaling. This structures our current multidisciplinary knowledge and views of the mechanisms governing the coupling of recognition to signal transduction and cell response. Importantly, this approach not only reveals previously unrecognized striking similarities in the basic mechanistic principles of function of numerous functionally diverse and unrelated surface membrane receptors, but also suggests the similarity between therapeutic targets, thus opening new horizons for both fundamental and clinically relevant studies.


High harmonic emission from a superposition of multiple unrelated frequency fields.
We report observations and analysis of high harmonic generation driven by a superposition of fields at 1290 nm and 780 nm. These fields are not commensurate in frequency and the superposition leads to an increase in the yield of the mid-plateau harmonics of more than two orders of magnitude compared to using the 1290 nm field alone. Significant extension of the cut-off photon energy is seen even by adding only a small amount of the 780 nm field. These observations are explained by calculations performed in the strong field approximation. Most importantly we find that enhancement is found to arise as a consequence of both increased ionization in the sum-field and modification of the electron trajectories leading to an earlier return time. The enhanced yield even when using modest intensity fields of 5 x 10(13) Wcm(-2) is extended to the 80 eV range and is a promising route to provide a greater photon number for applications in XUV imaging and time-resolved experiments at a high repetition rate.


Risk of subarachnoid haemorrhage according to number of affected relatives: a population based case-control study.
Relatives of patients with aneurysmal subarachnoid haemorrhage (SAH) have an increased risk of this type of stroke. In a population-based study, we analysed individualized risks of SAH according to the number of affected first-degree relatives. We retrieved all patients diagnosed with SAH in 2001-05 from the Swedish Inpatient Register. For each of the 5,282 patients, we identified five controls (n = 26,402) through the nationwide Register of Total Population. Through the Multi-generation Register, we retrieved all first-degree relatives for patients and controls and checked whether these 130,373 relatives had been diagnosed with SAH. By means of conditional logistic regression, we calculated odds ratios with corresponding 95% confidence intervals (95% CI) for the risk of SAH according to the number of affected relatives, and to the gender, age and type of kinship of the patient and affected relative. The odds ratio of SAH for individuals with one affected first-degree relative was 2.15 (95% CI 1.77-2.59). For individuals with two affected first-degree relatives, the odds ratio was 51.0 (95% CI 8.56-1117). Gender, age and type of kinship did not influence the risk for individuals with one or more affected relatives. The risk of SAH is slightly increased in the cases with one, but strongly increased in cases with two or more affected first-degree relatives. The latter strongly increased risk corresponds to a considerable absolute life-time risk of SAH and underscores the need to consider screening for aneurysms in these individuals.


Epidermal growth factor stimulates chloride transport in primary cultures of weanling and adult rabbit colonocytes.
We have shown that Ca2+-dependent regulation of Cl- secretion in the mammalian colon exhibits age dependence. Because epidermal growth factor (EGF) has a well-established role in growth and can increase intracellular calcium [Ca2+]i, it is conceivable that its developmental influence may extend to the regulation of intestinal ion transport. In this study, we examined the role of EGF in the regulation of Cl- transport in the developing rabbit distal colon. Because serum contains growth factors, which could have confounded our studies, we first established an optimal milieu for testing EGF in primary cultures of adult rabbit distal colonocytes by culturing them for 24 h in media containing 0%, 1%, 5%, and 20% serum. Chloride transport (millimoles per second) and [Ca2+]i were measured with use of the fluorescent indicator N-(ethoxycarbonylmethyl)-6-methoxyquinolinium bromide (MQAE) and Fura-2AM, respectively. Serum depletion had no effect on cell number, DNA content, or basal Cl- transport, but it significantly affected cell viability. In media with 0%, 1%, or 20% serum, bethanechol, 8BrcAMP, taurodeoxycholate, and EGF stimulated Cl- transport to a similar extent. EGF maximally stimulated Cl- transport at 16.3 nmol/L and 20 minutes. Bethanechol, but not EGF, increased [Ca2+]i. EGF did not alter bethanechol-stimulated Cl- transport or [Ca2+]i. EGF acts via an EGF-receptor and mitogen activated protein kinase (MAPK) signaling pathway, since stimulation of Cl- transport was abolished by genistein, AG1478, and PD98059. Weanling and adult colonocytes, cultured in 1% serum, showed similar basal and EGF-stimulated Cl- transport. EGF stimulates rabbit colonic Cl- transport via a Ca2+-independent, tyrosine kinase- and MAPK-dependent pathway, and its effects are not age dependent.


[The measurement of methane dissolved in water using raman spectroscopy assisted with CCl4 extraction].
While under laboratory conditions, the concentration of methane dissolved in water is too low to be detected because of the low solubility of methane using Raman spectroscopy. In the present paper, a novel approach based on CCl4 extraction was introduced, and used in the measurement of methane dissolved in water using Raman spectroscopy under laboratory conditions. Saturated aqueous solution of CH4, CCl4 solution after extraction of CH4 from the saturated aqueous solution and the saturated CCl4 solution of CH4 were prepared, and the Raman spectra of three samples were obtained. The obtained results show that the CH4 dissolved in saturated aqueous solution(the concentration of CH4 is about 1.14 mmol x L(-1)) can't been detected using Raman spectroscopy under laboratory conditions, but the CH4 Raman peak can be clearly seen in the spectra obtained for CCl4 solution after extraction. All the results demonstrate that the proposed approach can improve the Raman spectroscopy sensitivity of methane detection dissolved in water, and this approach has significant potential to be developed as an effective method for detection of methane dissolved in water.


Approaching a diagnostic point-of-care test for pediatric tuberculosis through evaluation of immune biomarkers across the clinical disease spectrum.
The World Health Organization (WHO) calls for an accurate, rapid, and simple point-of-care (POC) test for the diagnosis of pediatric tuberculosis (TB) in order to make progress "Towards Zero Deaths". Whereas the sensitivity of a POC test based on detection of Mycobacterium tuberculosis (MTB) is likely to have poor sensitivity (70-80% of children have culture-negative disease), host biomarkers reflecting the on-going pathological processes across the spectrum of MTB infection and disease may hold greater promise for this purpose. We analyzed transcriptional immune biomarkers direct ex-vivo and translational biomarkers in MTB-antigen stimulated whole blood in 88 Indian children with intra-thoracic TB aged 6 months to 15 years, and 39 asymptomatic siblings. We identified 12 biomarkers consistently associated with either clinical groups "upstream" towards culture-positive TB on the TB disease spectrum (CD14, FCGR1A, FPR1, MMP9, RAB24, SEC14L1, and TIMP2) or "downstream" towards a decreased likelihood of TB disease (BLR1, CD3E, CD8A, IL7R, and TGFBR2), suggesting a correlation with MTB-related pathology and high relevance to a future POC test for pediatric TB. A biomarker signature consisting of BPI, CD3E, CD14, FPR1, IL4, TGFBR2, TIMP2 and TNFRSF1B separated children with TB from asymptomatic siblings (AUC of 88%).


Accessory spleen-like masses in oncology patients: Are they always benign?
To assess retrospectively the significance of accessory spleen-like mass (ASLM) in oncology patients undergoing positron emission tomography/computed tomography (PET/CT). The results of PET/CT of 913 patients (278 lymphoma; 635 solid tumors) were reviewed. The number, size, location and attenuation of all ASLMs, and spleen attenuation, were recorded. ASLM fluorodeoxyglucose uptake was graded as normal (less than or equal to that in the liver) or representative of malignancy (more than in the liver). Follow-up PET/CT in patients with ASLM was reviewed when available. ASLM size and attenuation for spleen and ASLM were compared by unpaired Student's t test. The χ(2) and Fisher's exact tests were used to compare ASLM frequency and uptake for lymphomatous and solid tumors, respectively. ASLM frequency was 14.8%, with 152 ASLMs found in 135 patients. Mean attenuation was lower in ASLM compared with spleen by enhanced and non-enhanced CT (80.7 ± 20.4 HU vs 92.0 ± 14.4 HU, P < 0.0011 and 42.3 ± 9.0 HU vs 51.5 ± 6.3 HU, P < 0.0001, respectively). ASLM incidence was higher in lymphoma patients (56/278, 20.1%) than in those with solid tumors (56/278, 20.1% vs 79/635, 12.4%, P = 0.0036). Pathological uptake was found in four (7.1%) lymphoma patients but not in any patients with a solid tumor (P = 0.028) and it upstaged one patient with lymphoma. Follow-up PET/CT within 3-16 mo was available in 54% of patients with ASLM. Lesion regression was noted in all four pathological ASLMs on follow-up PET/CT after chemotherapy. In patients with lymphoma, ASLM can represent malignancy, and thus further characterization with PET/CT might be warranted. Patients with neoplasia other than ASLM can be confidently diagnosed with accessory spleen.


Peripheral neuropathy in children and adolescents treated for cancer.
Peripheral neuropathy is a well recognised treatment-related toxicity in children with cancer, associated with exposure to neurotoxic chemotherapy agents. Acute damage can occur in sensory, motor, or autonomic neurons, with symptoms that are rarely life threatening, but often severe enough to interfere with function during therapy and after treatment ends. The type of neuropathy and specific symptoms are associated with multiple factors including age at time of therapy, genetic predisposition, chemotherapy type and cumulative dose, and exposure to other agents during therapy. In this Review, we describe the peripheral neuropathy phenotype in children during cancer therapy and among survivors who have completed therapy, to summarise genetic and treatment-related risk factors for neuropathy, and to outline strategies to monitor and detect neuropathy during and after therapy. Additionally, we outline strategies for medical management of neuropathy during treatment and potential rehabilitation interventions to prevent or remediate functional loss.


Metabolism of excess methionine in the liver of intact rat: an in vivo 2H NMR study.
L-Methionine is the most toxic amino acid if supplied in excess, and the metabolic basis for this toxicity has been extensively studied, with varying conclusions. It is demonstrated here that in vivo 2H NMR spectroscopy provides a useful approach to the study of the hepatic metabolism of methionine in the anesthetized rat. Resonances corresponding to administered L-[methyl-2H3]methionine, and to the transmethylation product sarcosine, are observed during the first 10-min period after an intravenous injection of the labeled methionine, and the time dependence has been followed for a period of 5 h. A third resonance, assigned to the N-trimethyl groups of carnitine, phosphorylcholine, and other metabolites, becomes observable several hours after administration of the deuteriated methionine. In addition, there is a small increase in the intensity of the HDO resonance over the period of the study, which is interpreted to reflect the ultimate oxidation of the labeled sarcosine methyl group via mitochondrial sarcosine dehydrogenase. Additional small 2H resonances assigned to N1-methylhistidine and creatine could be observed in perchloric acid extracts of the livers of rats treated with the deuteriated methionine. Inhibition of the flux through the transmethylation pathway is observed in the rat pretreated with the S-ethyl analogue of methionine, ethionine. These data provide strong support for the importance of glycine transmethylation in the catabolism of excess methionine.


Preschoolers with autism show greater impairment in receptive compared with expressive language abilities.
In early typical language development, children understand words before they are able to use them in speech. Children with autism spectrum disorders (ASD) generally show impairments in both the comprehension and the production of language. However, the relative degree of delay or impairment in each of these sub-domains may also be atypical and remains less well-understood. Relative delay in receptive and expressive language skills was examined within a large sample of preschoolers with autism. Children's language abilities varied from pre-verbal to fluent speech. Scores on one direct clinician assessment and two parent-report measures of language were obtained for 152 preschoolers with core autism. As expected, on average, the language ability of the children with autism was lower than typical age norms, albeit with substantial individual variability. On all three language measures, receptive ability was relatively more impaired than expressive ability. Higher non-verbal ability was associated with such an atypical language profile. Recognition of the marked receptive language impairment relative to expressive language, found to affect at least one-third of preschoolers with autism in this sample, has important implications for interacting with these children and for informing appropriate targets in language and communication intervention.


A Staphylococcus aureus ypfP mutant with strongly reduced lipoteichoic acid (LTA) content: LTA governs bacterial surface properties and autolysin activity.
Many Gram-positive bacteria produce lipoteichoic acid (LTA) polymers whose physiological roles have remained a matter of debate because of the lack of LTA-deficient mutants. The ypfP gene responsible for biosynthesis of a glycolipid found in LTA was deleted in Staphylococcus aureus SA113, causing 87% reduction of the LTA content. Mass spectrometry and nuclear magnetic resonance spectroscopy revealed that the mutant LTA contained a diacylglycerol anchor instead of the glycolipid, whereas the remaining part was similar to the wild-type polymer except that it was shorter. The LTA mutant strain revealed no major changes in patterns of cell wall proteins or autolytic enzymes compared with the parental strain indicating that LTA may be less important in S. aureus protein attachment than previously thought. However, the autolytic activity of the mutant was strongly reduced demonstrating a role of LTA in controlling autolysin activity. Moreover, the hydrophobicity of the LTA mutant was altered and its ability to form biofilms on plastic was completely abrogated indicating a profound impact of LTA on physicochemical properties of bacterial surfaces. We propose to consider LTA and its biosynthetic enzymes as targets for new antibiofilm strategies.


[Relative biological effectiveness of tritium oxide based on nucleic acid metabolic indices].
Relative biological effectiveness (RBE) of tritium oxide, as compared to gamma-rays (137Cs), with regard to LD50/30 is 2,32 +/- 0,69 for rats. The RBE coefficients for tritium oxide are obtained with regard to some indices of nucleic acid metabolism in the thymus and spleen during the dose formation (0-14 days). The RBE of tritium oxide increases with a decrease in radiation dose as determined according to the concentration and content of DNA per organ and activity of thymus DNAases.


Parents in Prison: a child abuse and neglect prevention strategy.
Parents in Prison is an innovative family support service housed at the Tennessee State Prison for Men. Developed in response to inmates' recognition of the need for child abuse and neglect prevention services relevant to men who are in prison, the program model has potential for becoming a major service delivery strategy. Leadership for Parents in Prison is provided by an inmate committee which receives consultation and assistance from a community advisory board and an institutional sponsor. Program components--correspondence and classroom courses, monthly events featuring community guest speakers, and family-focused social activities and projects--address family needs during incarceration and upon return to community living. These diverse components and the flexible service delivery format permit widespread inmate participation not readily available in a prison setting. Parents in Prison successes demonstrate that the period of incarceration can be used to improve parental skills and knowledge and to strengthen family relationships. They also demonstrate the viability of a family support service which relies on inmate leadership, community volunteer participation, and institutional support. The future success of this model depends on replication in other prison settings, dissemination of the program products which have been and continue to be developed, and rigorous, systematic examination of the impact that participation has on child abuse and neglect problems associated with a father's incarceration.


Hip resurfacing arthroplasty complicated by mismatched implant components.
Metal-on-metal hip resurfacing has gained popularity as a feasible treatment option for young and active patients with hip osteoarthritis and high functional expectations. This procedure should only be performed by surgeons who have trained specifically in this technique. Preoperative planning is essential for hip resurfacing in order to execute a successful operation and preview any technical problems. The authors present a case of a man who underwent a resurfacing arthroplasty for osteoarthritis of the left hip that was complicated by mismatched implant components that were revised three days afterwards for severe pain and leg length discrepancy. Such mistakes, although rare, can be prevented by educating operating room staff in the size and colour code tables provided by the companies on their prostheses or implant boxes.


Functional epialleles at an endogenous human centromere.
Human centromeres are defined by megabases of homogenous alpha-satellite DNA arrays that are packaged into specialized chromatin marked by the centromeric histone variant, centromeric protein A (CENP-A). Although most human chromosomes have a single higher-order repeat (HOR) array of alpha satellites, several chromosomes have more than one HOR array. Homo sapiens chromosome 17 (HSA17) has two juxtaposed HOR arrays, D17Z1 and D17Z1-B. Only D17Z1 has been linked to CENP-A chromatin assembly. Here, we use human artificial chromosome assembly assays to show that both D17Z1 and D17Z1-B can support de novo centromere assembly independently. We extend these in vitro studies and demonstrate, using immunostaining and chromatin analyses, that in human cells the centromere can be assembled at D17Z1 or D17Z1-B. Intriguingly, some humans are functional heterozygotes, meaning that CENP-A is located at a different HOR array on the two HSA17 homologs. The site of CENP-A assembly on HSA17 is stable and is transmitted through meiosis, as evidenced by inheritance of CENP-A location through multigenerational families. Differences in histone modifications are not linked clearly with active and inactive D17Z1 and D17Z1-B arrays; however, we detect a correlation between the presence of variant repeat units of D17Z1 and CENP-A assembly at the opposite array, D17Z1-B. Our studies reveal the presence of centromeric epialleles on an endogenous human chromosome and suggest genomic complexities underlying the mechanisms that determine centromere identity in humans.


A conserved PTEN/FOXO pathway regulates neuronal morphology during C. elegans development.
The phosphatidylinositol 3-kinase (PI3K) signaling pathway is a conserved signal transduction cascade that is fundamental for the correct development of the nervous system. The major negative regulator of PI3K signaling is the lipid phosphatase DAF-18/PTEN, which can modulate PI3K pathway activity during neurodevelopment. Here, we identify a novel role for DAF-18 in promoting neurite outgrowth during development in Caenorhabditis elegans. We find that DAF-18 modulates the PI3K signaling pathway to activate DAF-16/FOXO and promote developmental neurite outgrowth. This activity of DAF-16 in promoting outgrowth is isoform-specific, being effected by the daf-16b isoform but not the daf-16a or daf-16d/f isoform. We also demonstrate that the capacity of DAF-16/FOXO in regulating neuron morphology is conserved in mammalian neurons. These data provide a novel mechanism by which the conserved PI3K signaling pathway regulates neuronal cell morphology during development through FOXO.


Effects of anticoagulant, serum, and temperature on the natural killer activity of human peripheral blood mononuclear cells stored overnight.
The effects of immediate versus delayed cell separation, storage temperature, presence of serum, and type of anticoagulation on the natural killer (NK) cytotoxicity of human mononuclear cells were assessed. The NK cytotoxicity of Ficoll-Hypaque-separated peripheral blood mononuclear cells (PBMC) was tested in a 3-h chromium-51 release assay with K562 cells at various effector/target cell ratios. The NK activities of PBMC from blood anticoagulated with either heparin or EDTA and then immediately separated and assayed were not different (42.9 +/- 2.5% for heparin and 40.3 +/- 4.6% for EDTA). When these separated cells were cultured in medium with 10% fetal calf serum and stored at 4,25, or 37 degrees C for 18 h before the assay, there was a significant increase in cytotoxicity. PBMC from blood stored in heparin or EDTA for 18 h before separation had reduced NK cytotoxicity, particularly if they were kept at 37 degrees C. When separated PBMC were cultured in medium with 10% human AB serum, however, samples held at 25 and 37 degrees C decreased in cytotoxicity but samples held at 4 degrees C maintained the cytotoxicity demonstrated at the baseline level with fresh cells. We recommend that heparinized blood be used for NK assays and that the PBMC be isolated immediately and held overnight at 4 degrees C in medium with 10% AB serum if the assay must be delayed. The NK cytotoxicity under these storage conditions most closely matches the results obtained when the PBMC are isolated and tested on the same day. IF PBMC isolation must also be postponed, it is best to store the blood in heparinized tubes at 25 degrees C to prevent loss of cytotoxic function.


Effector T-cell infiltration positively impacts survival of glioblastoma patients and is impaired by tumor-derived TGF-β.
In glioma-in contrast to various other cancers-the impact of T-lymphocytes on clinical outcome is not clear. We investigated the clinical relevance and regulation of T-cell infiltration in glioma. T-cell subpopulations from entire sections of 93 WHO°II-IV gliomas were computationally identified using markers CD3, CD8, and Foxp3; survival analysis was then done on primary glioblastomas (pGBM). Endothelial cells expressing cellular adhesion molecules (CAM) were similarly computationally quantified from the same glioma tissues. Influence of prominent cytokines (as measured by ELISA from 53 WHO°II-IV glioma lysates) on CAM-expression in GBM-isolated endothelial cells was determined using flow cytometry. The functional relevance of the cytokine-mediated CAM regulation was tested in a transmigration assay using GBM-derived endothelial cells and autologous T-cells. Infiltration of all T-cell subsets increased in high-grade tumors. Most strikingly, within pGBM, elevated numbers of intratumoral effector T cells (T(eff), cytotoxic and helper) significantly correlated with a better survival; regulatory T cells were infrequently present and not associated with GBM patient outcome. Interestingly, increased infiltration of T(eff) cells was related to the expression of ICAM-1 on the vessel surface. Transmigration of autologous T cells in vitro was markedly reduced in the presence of CAM-blocking antibodies. We found that TGF-β molecules impeded transmigration and downregulated CAM-expression on GBM-isolated endothelial cells; blocking TGF-β receptor signaling increased transmigration. This study provides comprehensive and novel insights into occurrence and regulation of T-cell infiltration in glioma. Specifically, targeting TGF-β1 and TGF-β2 might improve intratumoral T-cell infiltration and thus enhance effectiveness of immunotherapeutic approaches.


[Complications of plastic and reconstructive operations on the large intestine and their prevention].
The paper analyzes the complex clinico-experimental studies carried out by the surgeons, endoscopist, roentgenologist, and morphologist. Under special attention were the immediate and the late results of such variants of plastic and reconstructive operations on the large intestine when the remaining right portions of the large intestine were used for the substitution of defects of the left ones (coloplasty). Per 120 plastic and reconstructive operations there were 38 complications with 5 lethal outcomes (4,1%). The typical complications after coloplasty are described and means for their prevention are proposed.


Initial and Sustained Response Effects of 3 Vagal Maneuvers in Supraventricular Tachycardia: A Randomized, Clinical Trial.
For acute termination of supraventricular tachycardia (SVT), vagal maneuvers, including the standard Valsalva maneuver (sVM), modified Valsalva (mVM) maneuver, and carotid sinus massage (CSM), are first-line interventions. There is no criterion standard technique. This prospective, randomized study was aimed at analyzing the success rates of these 3 vagal maneuvers as measured by sustaining sinus rhythm at the fifth minute and SVT termination. We conducted this prospective, randomized controlled study in an emergency department (ED). We enrolled all the patients who were admitted to the ED and diagnosed with SVT. We randomly assigned them to 3 groups receiving sVM, mVM, and CSM and recorded the patients' responses to the vagal maneuvers and SVT recurrence after vagal maneuvers. The study was completed with 98 patients. A total of 25 (25.5%) instances of SVT were initially treated successfully with vagal maneuvers. The success rate was 43.7% (14/32 cases) from mVM, 24.2% (8/33) for sVM, and 9.1 % (3/33) for CSM (p < 0.05). At the end of the fifth minute, only 12.2% (12/98) of all patients had sinus rhythm. Sinus rhythm persisted in 28.1% (9/32) of patients in the mVM group, 6.1% (2/33) of patients in the sVM group, and 3% (1/33) in the CSM group at the fifth minute (p < 0.05). mVM is superior to the CSM maneuver in terminating SVT and maintaining rhythm. We conclude that it is beneficial to use mVM, which is more effective and lacks side effects.


Preliminary PET Study of 18F-FC119S in Normal and Alzheimer's Disease Models.
To evaluate the efficacy of 18F-FC119S as a positron emission tomography (PET) radiopharmaceutical for the imaging of Alzheimer's disease (AD), we studied the drug absorption characteristics and distribution of 18F-FC119S in normal mice. In addition, we evaluated the specificity of 18F-FC119S for β-amyloid (Aβ) in the AD group of an APP/PS1 mouse model and compared it with that in the wild-type (WT) group. The behavior of 18F-FC119S in the normal mice was characteristic of rapid brain uptake and washout patterns. In most organs, including the brain, 18F-FC119S reached its maximum concentration within 1 min and was excreted via the intestine. Brain PET imaging of 18F-FC119S showed highly specific binding of the molecule to Aβ in the cortex and hippocampus. The brain uptake and binding values for the AD group were higher than those for the WT group. These results indicated that 18F-FC119S would be a candidate PET imaging agent for targeting Aβ plaque.


Stage 4 s neuroblastoma: features, management and outcome of 268 cases from the Italian Neuroblastoma Registry.
Infants diagnosed with stage 4 s neuroblastoma commonly experience spontaneous disease regression, with few succumbing without response to therapy. We analyzed a large cohort of such infants enrolled in the Italian Neuroblastoma Registry to detect changes over time in presenting features, treatment and outcome. Of 3355 subjects aged 0-18 years with previously untreated neuroblastoma diagnosed between 1979 and 2013, a total of 280 infants (8.3%) had stage 4 s characteristics, 268 of whom were eligible for analyses. Three treatment eras were identified on the basis of based diagnostic and chemotherapy adopted. Group 1 patients received upfront chemotherapy; Group 2 and 3 patients underwent observation in the absence of life-threatening symptoms (LTS), except for Group 3 patients with amplified MYCN gene, who received more aggressive therapy. The three groups were comparable, with few exceptions. Ten-year overall survival significantly increased from 76.9 to 89.7% and was worse for male gender, age 0-29 days and presence of selected LTS on diagnosis, elevated LDH, and abnormal biologic features. Infants who underwent primary resection ± chemotherapy did significantly better. On multivariate analysis, treatment eras and the association of hepatomegaly to dyspnea were independently associated with worse outcome. Our data confirm that stage 4 s neuroblastoma is curable in nearly 90% of cases. Hepatomegaly associated to dyspnea was the most important independent risk factor. The cure rate could be further increased through timely identification of patients at risk who might benefit from surgical techniques, such as intra-arterial chemoembolization and/or liver transplantation, which must be carried out in institutions with specific expertise.


[Current problems in the formation of a system of monitoring health status of children and adolescents].
Discusses obviously limited potentialities of obtaining systematized analytical data by organs and institutions of social sphere. Such potentialities are particularly low in public health, specifically pediatric research and practice, where analytical materials would help improve the development of strategic programs of maternity and childhood protection and adopt adequate managing solutions at all levels. One of the constructive decisions aimed at the formation of a system of sociohygienic monitoring of children's health and ensuring integration of efforts of all responsible organs is creation of Federal Center of Social prevention for Children and Adolescents. The Center will ensure economy and rational utilization of finances and staff at the state and territorial level. Its purpose is to unite the interests of all profile organs and institutions for ensuring social well-being of children and adolescents.


Performance of ultra-high-frequency ultrasound in the evaluation of skin involvement in systemic sclerosis: a preliminary report.
High frequency ultrasound allows visualization of epidermis, dermis and hypodermis, precise measurement of skin thickness, as well as assessment of skin oedema, fibrosis and atrophy. The aim of this pilot cross-sectional observational study was to assess the performance and multiobserver variability of ultra-high-frequency (UHF) (50 MHz) ultrasound (US) in measuring skin thickness as well as the capacity of UHF-derived skin features to differentiate SSc patients from healthy controls. Twenty-one SSc patients (16 limited and five diffuse SSc) and six healthy controls were enrolled. All subjects underwent US evaluation by three experts at three anatomical sites (forearm, hand and finger). Dermal thickness was measured and two rectangular regions of interest, one in dermis and one in hypodermis, were established for texture feature analysis. UHF-US allowed a precise identification and measurement of the thickness of the dermis. The dermal thickness in the finger was significantly higher in patients than in controls (P < 0.05), while in the forearm it was significantly lower in patients than in controls (P < 0.001). Interobserver variability for dermal thickness was good to excellent [forearm intraclass correlation coefficient (ICC) = 0.754; finger ICC = 0.699; hand ICC = 0.602]. Texture computed analysis of dermis and hypodermis was able to discriminate between SSc and healthy subjects (area under the curve >0.7). These preliminary data show that skin UHF-US allows a very detailed imaging of skin layers, a reliable measurement of dermal thickness, and a discriminative capacity between dermis and hypodermis texture features in SSc and healthy subjects.


Assessing participation in a community-based health planning and services programme in Ghana.
Community participation is increasingly seen as a pre-requisite for successful health service uptake. It is notoriously difficult to assess participation and little has been done to advance tools for the assessment of community participation. In this paper we illustrate an approach that combines a 'social psychology of participation' (theory) with 'spider-grams' (method) to assess participation and apply it to a Community-based Health Planning and Services (CHPS) programme in rural Ghana. We draw on data from 17 individual in-depth interviews, two focus group discussions and a community conversation with a mix of service users, providers and community health committee members. It was during the community conversation that stakeholders collectively evaluated community participation in the CHPS programme and drew up a spider-gram. Thematic analysis of our data shows that participation was sustained through the recognition and use of community resources, CHPS integration with pre-existing community structures, and alignment of CHPS services with community interests. However, male dominance and didactic community leadership and management styles undermined real opportunities for broad-based community empowerment, particularly of women, young people and marginalised men. We conclude that combining the 'spider-gram' tool and the 'social psychology of participation' framework provide health professionals with a useful starting point for assessing community participation and developing recommendations for more participatory and empowering health care programmes.


Protection of synapses against Alzheimer's-linked toxins: insulin signaling prevents the pathogenic binding of Abeta oligomers.
Synapse deterioration underlying severe memory loss in early Alzheimer's disease (AD) is thought to be caused by soluble amyloid beta (Abeta) oligomers. Mechanistically, soluble Abeta oligomers, also referred to as Abeta-derived diffusible ligands (ADDLs), act as highly specific pathogenic ligands, binding to sites localized at particular synapses. This binding triggers oxidative stress, loss of synaptic spines, and ectopic redistribution of receptors critical to plasticity and memory. We report here the existence of a protective mechanism that naturally shields synapses against ADDL-induced deterioration. Synapse pathology was investigated in mature cultures of hippocampal neurons. Before spine loss, ADDLs caused major downregulation of plasma membrane insulin receptors (IRs), via a mechanism sensitive to calcium calmodulin-dependent kinase II (CaMKII) and casein kinase II (CK2) inhibition. Most significantly, this loss of surface IRs, and ADDL-induced oxidative stress and synaptic spine deterioration, could be completely prevented by insulin. At submaximal insulin doses, protection was potentiated by rosiglitazone, an insulin-sensitizing drug used to treat type 2 diabetes. The mechanism of insulin protection entailed a marked reduction in pathogenic ADDL binding. Surprisingly, insulin failed to block ADDL binding when IR tyrosine kinase activity was inhibited; in fact, a significant increase in binding was caused by IR inhibition. The protective role of insulin thus derives from IR signaling-dependent downregulation of ADDL binding sites rather than ligand competition. The finding that synapse vulnerability to ADDLs can be mitigated by insulin suggests that bolstering brain insulin signaling, which can decline with aging and diabetes, could have significant potential to slow or deter AD pathogenesis.


The allocation of ecosystem net primary productivity in tropical forests.
The allocation of the net primary productivity (NPP) of an ecosystem between canopy, woody tissue and fine roots is an important descriptor of the functioning of that ecosystem, and an important feature to correctly represent in terrestrial ecosystem models. Here, we collate and analyse a global dataset of NPP allocation in tropical forests, and compare this with the representation of NPP allocation in 13 terrestrial ecosystem models. On average, the data suggest an equal partitioning of allocation between all three main components (mean 34 ± 6% canopy, 39 ± 10% wood, 27 ± 11% fine roots), but there is substantial site-to-site variation in allocation to woody tissue versus allocation to fine roots. Allocation to canopy (leaves, flowers and fruit) shows much less variance. The mean allocation of the ecosystem models is close to the mean of the data, but the spread is much greater, with several models reporting allocation partitioning outside of the spread of the data. Where all main components of NPP cannot be measured, litterfall is a good predictor of overall NPP (r(2) = 0.83 for linear fit forced through origin), stem growth is a moderate predictor and fine root production a poor predictor. Across sites the major component of variation of allocation is a shifting allocation between wood and fine roots, with allocation to the canopy being a relatively invariant component of total NPP. This suggests the dominant allocation trade-off is a 'fine root versus wood' trade-off, as opposed to the expected 'root-shoot' trade-off; such a trade-off has recently been posited on theoretical grounds for old-growth forest stands. We conclude by discussing the systematic biases in estimates of allocation introduced by missing NPP components, including herbivory, large leaf litter and root exudates production. These biases have a moderate effect on overall carbon allocation estimates, but are smaller than the observed range in allocation values across sites.


A multidimensional model of fatigue in patients with rheumatoid arthritis.
To evaluate a multidimensional model testing disease activity, mood disturbance, and poor sleep quality as determinants of fatigue in patients with rheumatoid arthritis (RA). The data of 106 participants were drawn from baseline of a randomized comparative efficacy trial of psychosocial interventions for RA. Sets of reliable and valid measures were used to represent model constructs. Structural equation modeling was used to test the direct effects of disease activity, mood disturbance, and poor sleep quality on fatigue, as well as the indirect effects of disease activity as mediated by mood disturbance and poor sleep quality. The final model fit the data well, and the specified predictors explained 62% of the variance in fatigue. Higher levels of disease activity, mood disturbance, and poor sleep quality had direct effects on fatigue. Disease activity was indirectly related to fatigue through its effects on mood disturbance, which in turn was related to poor sleep quality. Mood disturbance also indirectly influenced fatigue through poor sleep quality. Our findings confirmed the importance of a multidimensional framework in evaluating the contribution of disease activity, mood disturbance, and sleep quality to fatigue in RA using a structural equation approach. Mood disturbance and poor sleep quality played major roles in explaining fatigue along with patient-reported disease activity.


A rapid method for determining tRNA charging levels in vivo: analysis of yeast mutants defective in the general control of amino acid biosynthesis.
We describe a simple method to quantitate the intracellular levels of charged tRNA species representing all 20 amino acids. Small RNA species are isolated from yeast cells under conditions where amino acids remain bound to their cognate tRNAs. After chromatographic removal of free amino acids, the tRNAs are discharged, and the amounts of the released amino acids are then quantitated. This method was applied to yeast cells from a wild type strain and from three mutant strains that are defective both in the general control of amino acid biosynthesis and in protein synthesis. Two of these mutant strains, previously shown to be defective in the methionine or isoleucine tRNA synthetases, respectively contain undetectable amounts of charged methionine or isoleucine although their levels of the remaining 19 amino acids are similar to a wild type strain. In contrast, a gcd1 mutant strain has normal levels of all 20 amino-acyl tRNA species. Thus, gcd1 strains are defective in general control of amino acid biosynthesis for reasons other than artifactual starvation of an amino acid due to a failure in tRNA changing.


Microvascular cerebral blood flow fluctuations in association with apneas and hypopneas in acute ischemic stroke.
In a pilot study on acute ischemic stroke (AIS) patients, unexpected periodic fluctuations in microvascular cerebral blood flow (CBF) had been observed. Motivated by the relative lack of information about the impact of the emergence of breathing disorders in association with stroke on cerebral hemodynamics, we hypothesized that these fluctuations are due to apneic and hypopneic events. A total of 28 patients were screened within the first week after stroke with a pulse oximeter. Five (18%) showed fluctuations of arterial blood oxygen saturation ( ≥ 3 % ) and were included in the study. Near-infrared diffuse correlation spectroscopy (DCS) was utilized bilaterally to measure the frontal lobe CBF alongside respiratory polygraphy. Biphasic CBF fluctuations were observed with a bilateral increase of 27.1 % ± 17.7 % and 29.0 % ± 17.4 % for the ipsilesional and contralesional hemispheres, respectively, and a decrease of - 19.3 % ± 9.1 % and - 21.0 % ± 8.9 % for the ipsilesional and contralesional hemispheres, respectively. The polygraph revealed that, in general, the fluctuations were associated with apneic and hypopneic events. This study motivates us to investigate whether the impact of altered respiratory patterns on cerebral hemodynamics can be detrimental in AIS patients.


Cellular expression and siRNA-mediated interference of rhodopsin cis-acting splicing mutants associated with autosomal dominant retinitis pigmentosa.
To investigate the cellular expression of cis-acting splicing mutations in the rhodopsin gene (RHO) that lead to autosomal dominant or recessive retinitis pigmentosa (adRP/arRP) and the role of nonsense-mediated mRNA decay (NMD) in its pathogenic mechanism. To design a potential therapeutic RNAi-based suppression strategy for cis-acting adRP splicing mutants. Cells were transfected with genomic constructs encoding the human wild-type (WT) and c.531-2A>G, c.936+1G>T, c.937-1G>T and c.745G>T RHO mutants. Total RNA was quantified by RT-PCR and protein was analyzed by immunocytochemistry. Three small interfering (si)RNAs directed against adRP mutant transcripts were designed and assayed in COS7 cells. The RHO cis-acting splicing mutations causing adRP, c.531-2A>G and c.937-1G>T, induce cryptic splicing. In contrast, the c.936+1G>T mutation, which causes arRP, results in exon skipping. Although the c.531-2A>G and c.745G>T RHO sequence predicted a premature termination codon (PTC) that should be a target for NMD, these mutant proteins were detected in transfected cells. The siRNAs designed to interfere with adRP mutants silenced the corresponding mRNA with varying efficiency. Although two RHO mutations that cause different RP phenotypes were the target for the NMD mechanism, a fraction of mutant RNA transcript may circumvent the NMD mechanism and be translated into protein. Thus, different levels of mutant protein may be necessary to trigger the RP phenotype. The findings demonstrate the potential use of siRNA to interfere with cis-acting splicing RHO transcripts. However, limitations in the mutation sequence and incomplete mutant transcript elimination should be considered in a therapeutic approach for adRP.


[A new class of N-methyl-D-aspartic acid receptor agonists and antagonists--derivatives of imidazole-4,5- and pyrazole-3,4-dicarboxylic acids].
New agonists and antagonists of the N-methyl-D-aspartic acid (NMDA) receptors were found among the derivatives of 1- or 2-alkyl-substituted imidazole-4,5- and pyrazole-3,4-dicarboxylic acids. Lipophilic surrounding of the nitrogen atom in these compounds was found to determine their ability to be either agonists or antagonists, while the distance between the terminal acidic functions was the same. An increase in the lipophilicity can also cause loss of selective action upon the NMDA receptors and occurrence of non- NMDA antagonistic activity.


Events Associated With Changes in Reliance on the Veterans Health Administration Among Medicare-eligible Veterans.
We can learn something about how Veterans value the Veterans Health Administration (VHA) versus community providers by observing Veterans' choices between VHA and Medicare providers after they turn 65. For a cohort of Veterans who were newly age-eligible for Medicare, we estimated the change in VHA reliance (VHA outpatient visits divided by total VHA and Medicare visits) associated with specific events: receiving a life-threatening diagnosis, having a Medicare-paid hospitalization, or moving further from the VHA. A longitudinal cohort study of VHA and Medicare administrative data. A total of 5932 VHA users who completed a health survey in 1999 and became age-eligible for Medicare from 1998 to 2000 were followed through 2016. More Veterans chose to rely on the VHA than Medicare (64% vs. 36.%). For a VHA-reliant Veteran, a Medicare-paid hospital stay was associated with a decrease of 7.8 percentage points (pps) (P<0.001) in VHA reliance in the subsequent 12 months, but by 36 months reliance increased to near prehospitalization levels (-1.5 pps; P=0.138). Moving further from the VHA, or receiving a diagnosis of cancer, heart failure, or renal failure had no significant association with subsequent VHA reliance; however, a diagnosis of dementia was associated with a decrease in VHA reliance (-8.6 pps; P=0.026). A significant majority of newly Medicare-eligible VHA users voted with their feet in favor of sustaining the VHA as a provider of comprehensive medical care for Veterans. These VHA-reliant Veterans maintained their reliance even after receiving a life-threatening diagnosis, and after experiencing Medicare-provided hospital care.


Effects of fish age versus size on the development of whirling disease in rainbow trout.
We examined the effects of both fish age and size on the development of resistance to whirling disease in Erwin strain rainbow trout. Previously, we demonstrated that juvenile rainbow trout became resistant to development of the disease when first exposed to triactinomyxons of the parasite Myxobolus cerebralis at about 9 wk post-hatch when raised at 12 degrees C, but ages and sizes of fish used in that experiment were confounded (Ryce EKN, Zale AV, MacConnell E [2004] Dis Aquat Org 59:225-233). In this study, rainbow trout of the same age and different sizes, and the same size and different ages, were exposed to the parasite to distinguish the influences of age and size. Fish were reared at 3 different water temperatures prior to exposure to produce groups with different growth rates and were exposed to the parasite at 7 or 9 wk post-hatch. Disease severity was affected by both age and size at first exposure, but the effects were not independent. An increase in fork length from 36 to 40 mm among fish exposed at 7 wk post-hatch did not confer increased resistance, but the same increase in size at 9 wk post-hatch did. Similarly, an increase in age from 7 to 9 wk post-hatch among fish exposed at 36 mm fork length did not confer increased resistance, but the same increase in age at 40 mm did. Rainbow trout must be both 9 wk post-hatch or older and at least 40 mm in fork length at time of exposure to exhibit enhanced resistance to whirling disease. Resistance to disease was not associated with the level of skeletal ossification.


The role of food allergy in atopic dermatitis.
Atopic dermatitis (AD) is a chronic, pruritic, inflammatory skin disease affecting more than 10% of all children. Sensitization to foods triggers isolated skin symptoms in about 30% of children. These symptoms include immediate reactions within minutes after ingesting food without exacerbation of AD and early and late exacerbations of AD. It is important to identify clinically relevant sensitizations to foods using skin prick tests, a specific IgE blood test (ImmunoCAP; Phadia, Portage, MI, USA), and double-blind, placebo-controlled food challenges to initiate appropriate dietary interventions and avoid unnecessary dietary restrictions. Children with AD triggered by food allergens demonstrate a distinct immune response upon stimulation of their peripheral blood mononuclear cells with food allergen. A defective skin barrier and increased intestinal permeability appear to facilitate allergen sensitization. Appropriate skin care to maintain skin barrier function and dietary avoidance of highly allergenic foods during infancy may help to prevent allergen sensitization, thereby reducing the severity of AD and food allergies.


Thrust plate prosthesis for proximal femoral deformity: a series of 15 patients.
Patients with coxarthrosis and proximal femoral deformity experience problems with total hip arthroplasty. A custom-made prosthesis or a proximal osteotomy is required for such cases, and these also increase the rate of complications. The purpose of this study was to evaluate the results of the thrust plate prosthesis (TPP) in patients with deformity of the proximal femur. Fifteen patients (7 females, 8 males) with a mean age of 56.4 years (range 19-75 years) at the time of the surgery were included in the study. The etiology was traumatic coxarthrosis in 12, and nonunion of a femoral neck fracture with osteonecrosis of the femoral head in the remaining three. While the femoral component was a third-generation TPP in all patients, the acetabular component was a Protek expansion cup in 12, and a cementless standard cup in three patients. All operations were performed through a Hardinge approach. Patients were followed up for at least 3 years (range 36-116 months) and evaluated clinically with the Harris Hip Score. The mean preoperative Harris Hip Score increased from 51.2 (range 15-79) to 92.7 (range 60-100) at the latest assessment. In two cases, loosening of the femoral component was observed in zone 3, both 12 months postoperatively. One was replaced by an intramedullary prosthesis, and the other was asymptomatic. TPP is a good alternative for patients with malformations of the proximal femur. The use of TPP avoids technical difficulties and a custom-made prosthesis.


Chitinases and chitinase-like proteins as biomarkers in neurologic disorders.
Chitinases are hydrolytic enzymes widely distributed in nature. Despite their physiologic and pathophysiologic roles are not well understood, chitinases are emerging as biomarkers in a broad range of neurologic disorders, where in many cases, protein levels measured in the CSF have been shown to correlate with disease activity and progression. In this review, we will summarize the structural features of human chitinases and chitinase-like proteins and their potential physiologic and pathologic functions in the CNS. We will also review existing evidence for the role of chitinases and chitinase-like proteins as diagnostic and prognostic biomarkers in inflammatory, neurodegenerative diseases, and psychiatric disorders. Finally, we will comment on future perspectives of chitinase studies in neurologic conditions.


[Electrophoretic analysis of the proteins of different alphavirus strains].
Proteins of various alphavirus strains: Venezuelan equine encephalomyelitis, Semliki Forest, and Sindbis, were studied by a high resolution polyacrylamide gel electrophoresis. In addition to structural C, E1 and E2 proteins, the infected cells were found to contain a number of nonstructural polypeptides: B, 83 kD (analogue of nsP2), 75 kD (nsP3), PE2, precursor of E2 and a product of modification of nucleocapsid protein C27. In virions, in addition to the main structural polypeptides, protein components with molecular weights about 100 kD were found which, most likely, were aggregates of E1/E2 glycoproteins, and 40 kD, a product of E1 protein degradation. The alphavirus strains under study differed both in the electrophoretic mobility of the above-mentioned virus-specific polypeptides and in the stability of B and PE2 proteins in the infected cells. The intracellular stability of polyprotein B depended considerably also on the host cell.


Deer antlers as a model of Mammalian regeneration.
Deer antlers are cranial appendages that develop after birth as extensions of a permanent protuberance (pedicle) on the frontal bone. Pedicles and antlers originate from a specialized region of the frontal bone; the 'antlerogeneic periosteum' and the systemic cue which triggers their development in the fawn is an increase in circulating androgen. These primary antlers are then shed and regenerated the following year in a larger, more complex form. Antler growth is extremely rapid-an adult red deer can produce a pair of antlers weighing approximately 30kg in three months, and involves both endochondral and intramembranous ossification. Since antlers are sexual secondary characteristics, their annual cycles of growth have evolved to be closely coordinated to the reproductive cycle which, in temperate species, is linked to the photoperiod. Cessation of antler growth and death of the overlying skin (velvet) coincides with a rise in circulating testosterone as the autumn breeding season approaches. The 'dead' antlers remain attached to the pedicle until they are shed (cast) the following spring when circulating testosterone levels fall. In red deer, the species that we study, casting of the old set of antlers is followed immediately by growth of the new set. Although the anatomy of antler growth and the endocrine changes associated with it have been well documented, the molecular mechanisms involved remain poorly understood. The case for continuing to decipher them remains compelling, despite the obvious limitations of using deer as an experimental model, because this research will help provide insight into why humans and other mammals have lost the ability to regenerate organs. From the information so far available, it would appear that the signaling pathways that control the development of skeletal elements are recapitulated in regenerating antlers. This apparent lack of any specific 'antlerogenic molecular machinery' suggests that the secret of deers' ability to regenerate antlers lies in the particular cues to which multipotential progenitor/stem cells in an antler's 'regeneration territory' are exposed. This in turn suggests that with appropriate manipulation of the environment, pluripotential cells in other adult mammalian tissues could be stimulated to increase the healing capacity of organs, even if not to regenerate them completely. The need for replacement organs in humans is substantial. The benefits of increasing individuals' own capacity for regeneration and repair are self evident.


A conservative approach for a patient with traumatically induced urinary incontinence.
To consider a conservative management plan for a patient suffering from diurnal enuresis. A 12-yr-old girl complained of left flank pain and involuntary urine loss that apparently began a short time after she fell backwards and landed on her coccyx. She had to wear a sanitary pad prophylactically throughout each day for more than a year because of unpredictable urinary incontinence. Manual adjusting procedures and soft tissue therapy were applied initially. Procedures were modified later to include an intrarectal technique. The quantity of urine loss decreased slowly with the initial treatment approach but never resolved completely. The flank pain also decreased somewhat but never ceased altogether. After the introduction of the intrarectal technique, additional progress was reported and both the flank pain and urinary incontinence resolved completely. A 4-yr follow-up confirmed complete resolution. Conservative care helped an adolescent girl suffering from traumatically induced urinary incontinence.


Comparative transcriptome analysis of leaves during early stages of chilling stress in two different chilling-tolerant brown-fiber cotton cultivars.
Chilling stress generates significant inhibition of normal growth and development of cotton plants and lead to severe reduction of fiber quality and yield. Currently, little is known for the molecular mechanism of brown-fiber cotton (BFC) to respond to chilling stress. Herein, RNA-sequencing (RNA-seq)-based comparative analysis of leaves under 4°C treatment in two different-tolerant BFC cultivars, chilling-sensitive (CS) XC20 and chilling-tolerant (CT) Z1612, was performed to investigate the response mechanism. A total of 72650 unigenes were identified with eight commonly used databases. Venn diagram analysis identified 1194 differentially expressed genes (DEGs) with significant up-regulation in all comparison groups. Furthermore, enrichment analyses of COG and KEGG, as well as qRT-PCR validation, indicated that 279 genes were discovered as up-regulated DEGs (UDEGs) with constant significant increased expression in CT cultivar Z1612 groups at the dimensions of both each comparison group and treatment time, locating in the enriched pathways of signal transduction, protein and carbohydrate metabolism, and cell component. Moreover, the comprehensive analyses of gene expression, physiological index and intracellular metabolite detections, and ascorbate antioxidative metabolism measurement validated the functional contributions of these identified candidate genes and pathways to chilling stress. Together, this study for the first time report the candidate key genes and metabolic pathways responding to chilling stress in BFC, and provide the effective reference for understanding the regulatory mechanism of low temperature adaptation in cotton.


Characterization of the core mammalian clock component, NPAS2, as a REV-ERBalpha/RORalpha target gene.
The mammalian clock is regulated at the cellular level by a transcriptional/translational feedback loop. BMAL1/clock (or NPAS2) heterodimers activate the expression of the period (PER) and cryptochrome (CRY) genes acting as transcription factors directed to the PER and CRY promoters via E-box elements. PER and CRY proteins form heterodimers and suppress the activity of the BMAL1/clock (or NPAS2) completing the feedback loop. The circadian expression of BMAL1 is influenced by retinoic acid receptor-related orphan receptor α (RORα) and REV-ERBα, two nuclear receptors that target a ROR-response element in the promoter of the BMAL1 gene. Given that BMAL1 functions as an obligate heterodimer with either clock or NPAS2, it is unclear how the expression of the partner is coordinated with BMAL1 expression. Here, we demonstrate that NPAS2 is also a RORα and REV-ERBα target gene. Using a ChIP/microarray screen, we identified both RORα and REV-ERBα occupancy of the NPAS2 promoter. We identified two functional ROREs within the NPAS2 promoter and also demonstrate that both RORα and REV-ERBα regulate the expression of NPAS2 mRNA. These data suggest a mechanism by which RORα and REV-ERBα coordinately regulate the expression of the positive arm of the circadian rhythm feedback loop.


Protection of plant varieties and parts as intellectual property.
In view of the Supreme Court decision in Chakrabarty v. Diamond, Commissioner of Patents and Trademarks, it is possible that plant varieties can be protected under three different U.S. statutes: the Plant Variety Protection Act, the Plant Patent Law, and the General Patent Law. The Plant Variety Protection Act protects varieties that are reproduced by seed, whereas the Plant Patent Law protects varieties reproduced asexually. Varieties, irrespective of how they are reproduced, could be patentable under the General Patent Statute. It is not clear whether parts of plants can be protected by grants under the Plant Patent Law or Plant Variety Protection Act and it is possible that they will be best protected under the General Patent Statute and by maintaining them as trade secrets. Only time will show whether the existing statutes are sufficient to provide both guidance and adequate protection or whether changes in the law will be required.


Association of a novel human immunodeficiency virus type 1 protease substrate cleft mutation, L23I, with protease inhibitor therapy and in vitro drug resistance.
We observed a previously uncharacterized mutation in the protease substrate cleft, L23I, in 31 of 4,303 persons undergoing human immunodeficiency virus type 1 genotypic resistance testing. In combination with V82I, L23I was associated with a sevenfold reduction in nelfinavir susceptibility and a decrease in replication capacity. In combination with other drug resistance mutations, L23I was associated with multidrug resistance and a compensatory increase in replication capacity.


Comparison of metabolic syndrome prevalence using eight different definitions: a critical approach.
Multiple definitions of the metabolic syndrome (MS) have been proposed for children, adolescents and adults. The aim of this study was to analyse the variations in the MS prevalence using different definitions and to examine which factors influence the frequency of the MS in childhood and adolescence. The prevalence of the MS according to eight proposed definitions was studied in 1205 Caucasian overweight children and adolescents aged 4-16 years (mean body mass index (BMI) 27.3 kg/m2, mean age 11.8 years, 46% males, 39% prepubertal). Blood pressure, waist circumference and fasting triglycerides, HDL-cholesterol, total cholesterol, insulin and glucose concentrations were determined. Overweight was defined according to the International Task Force of Obesity in Childhood. Degree of overweight was calculated as standard deviation score of BMI (SDS-BMI). Insulin resistance was estimated based on the HOMA model. The prevalence of the MS varied significantly (p<0.001), being between 6% and 39% depending on the different definitions. Only 2% of the children fulfilled the criteria of the MS in all definitions. Insulin resistance and degree of overweight were associated with the MS. In most definitions, pubertal stage did not influence the occurrence of the MS. In a principal component analysis, total cholesterol, triglycerides and waist circumference showed high final communality estimates. Since the prevalence of the MS varied widely in overweight children and adolescents depending on the proposed definition used, an internationally accepted uniform definition of the MS is necessary to compare different populations and studies.


How discordant morphological and molecular evolution among microorganisms can revise our notions of biodiversity on Earth.
Microscopy has revealed tremendous diversity of bacterial and eukaryotic forms. Recent molecular analyses show discordance in estimates of biodiversity between morphological and molecular analyses. Moreover, phylogenetic analyses of the diversity of microbial forms reveal evidence of convergence at scales as deep as interdomain: morphologies shared between bacteria and eukaryotes. Here, we highlight examples of such discordance, focusing on exemplary lineages such as testate amoebae, ciliates, and cyanobacteria. These have long histories of morphological study, enabling deeper analyses on both the molecular and morphological sides. We discuss examples in two main categories: (i) morphologically identical (or highly similar) individuals that are genetically distinct and (ii) morphologically distinct individuals that are genetically the same. We argue that hypotheses about discordance can be tested using the concept of neutral morphologies, or more broadly neutral phenotypes, as a null hypothesis.


Light-induced degeneration and microglial response in the retina of an epibenthonic pigmented teleost: age-dependent photoreceptor susceptibility to cell death.
Constant intense light causes apoptosis of photoreceptors in the retina of albino fish. However, very few studies have been performed on pigmented species. Tench (Tinca tinca) is a teleost inhabiting dimly lit environments that has a predominance of rods within the photoreceptor layer. To test the hypothesis that constant high intensity light can result in retinal damage in such pigmented epibenthonic teleost species, photodegeneration of the retina was investigated in the larvae and in juveniles of tench to assess whether any damage may also be dependent on fish age. We exposed both groups of animals to 5 days of constant darkness, followed by 4 days of constant 20,000 lx light, and then by 6 days of recovery in a 14 h light:10 h dark cycle. The results showed that the retina of the larvae group exhibited abundant photoreceptor cell apoptosis during the time of exposition to intense light, whereas that of juveniles was indifferent to it. Damaged retinas showed a strong TUNEL signal in photoreceptor nuclei, and occasionally a weak cytoplasmic TUNEL signal in Müller glia. Specific labelling of microglial cells with Griffonia simplicifolia lectin (GSL) histochemistry revealed that photoreceptor cell death alerts microglia in the degenerating retina, leading to local proliferation, migration towards the injured outer nuclear layer (ONL), and enhanced phagocytosis of photoreceptor debris. During the first days of intense light treatment, Müller cells phagocytosed dead photoreceptor cells but, once microglial cells became activated, there was a progressive increase in the phagocytic capacity of the microglia.


Comparison of image quality with 62Cu and 64Cu-labeled radiotracers in positron emission tomography whole-body phantom imaging.
PET imaging is possible with copper (Cu) isotopes, (60)Cu, (61)Cu, (62)Cu and (64)Cu. Although (62)Cu- and (64)Cu-labeled radiotracers are often used for preclinical and clinical PET studies, we do not know which radiotracers have better image quality for tumor imaging. In this study, we compare image quality between (62)Cu and (64)Cu imaging with a different acquisition mode and reconstruction algorithm using a whole-body phantom for tumor imaging. In a National Electrical Manufacturers Association (NEMA) 2001 whole-body phantom, the concentration of (62)Cu-ATSM and (64)Cu-ATSM was, respectively, approximately 2.7 and 1.8MBq/mL in all the spheres and approximately 0.9 and 0.6MBq/mL in the background. After adjustment for true coincidence events between (62)Cu and (64)Cu, two-dimensional (2D) and three-dimensional (3D) PET scan data were acquired for 10min. The data were reconstructed using filtered back projection (FBP) and the ordered subset expectation maximization (OSEM) algorithm. Image quality of (62)Cu and (64)Cu was compared using recovery coefficient (RC), sphere-to-background ratio (SBR) and coefficient of variation (%COV). There were little significant differences between (62)Cu and (64)Cu imaging, visually. Recovery coefficients of (64)Cu images were higher than those of (62)Cu images. The RC of (64)Cu images with 3D acquisition mode and OSEM was the highest in all experiments. No SBR values were significantly different from the true value of 3.0 in 37mm sphere diameters, but 3D acquisition and OSEM yielded slight overestimations compared with 2D acquisition and FBP, the gold standard for quantification in PET studies. Percentage COV values of (64)Cu with OSEM were significantly lower than those of (62)Cu. Copper-64 radiotracers provide higher image quality than (62)Cu-radiotracers in whole-body tumor imaging only when the 3D acquisition mode and OSEM algorithm are applied. However, the quantitative values for smaller tumors may be slightly overestimated.


Glutamate/GABA+ ratio is associated with the psychosocial domain of autistic and schizotypal traits.
The autism and schizophrenia spectra overlap to a large degree in the social and interpersonal domains. Similarly, abnormal excitatory glutamate and inhibitory γ-aminobutyric acid (GABA) neurotransmitter concentrations have been reported for both spectra, with the interplay of these neurotransmitters important for cortical excitation to inhibition regulation. This study investigates whether these neurotransmitter abnormalities are specific to the shared symptomatology, and whether the degree of abnormality increases with increasing symptom severity. Hence, the relationship between the glutamate/GABA ratio and autism and schizophrenia spectrum traits in an unmedicated, subclinical population was investigated. A total of 37 adults (19 female, 18 male) aged 18-38 years completed the Autism Spectrum Quotient (AQ) and Schizotypal Personality Questionnaire (SPQ), and participated in the resting state proton magnetic resonance spectroscopy study in which sequences specific for quantification of glutamate and GABA+ concentration were applied to a right and left superior temporal voxel. There were significant, moderate, positive relationships between right superior temporal glutamate/GABA+ ratio and AQ, SPQ and AQ+SPQ total scores (p<0.05), SPQ subscales Social Anxiety, No Close Friend, Constricted Affect, Odd Behaviour, Odd Speech, Ideas of Reference and Suspiciousness, and AQ subscales Social Skills, Communication and Attention Switching (p<0.05); increased glutamate/GABA+ coinciding with higher scores on these subscales. Only the relationships between glutamate/GABA+ ratio and Social Anxiety, Constricted Affect, Social Skills and Communication survived multiple comparison correction (p< 0.004). Left superior temporal glutamate/GABA+ ratio reduced with increasing restricted imagination (p<0.05). These findings demonstrate evidence for an association between excitatory/inhibitory neurotransmitter concentrations and symptoms that are shared between the autism and schizophrenia spectra.


Identification of clinically relevant viridans streptococci by analysis of transfer DNA intergenic spacer length polymorphism.
The utility of PCR analysis of transfer DNA intergenic spacer length polymorphism (tDNA-ILP) for the identification to the species level of clinically relevant viridans streptococci was evaluated with a collection of reference strains of 15 species of the salivarius, anginosus, mitis and mutans rRNA homology groups. PCR products generated by using fluorescent, outwardly directed, consensus tDNA primers were analysed by electrophoresis on denaturating polyacrylamide gels and by laser fluorescence scanning. Eleven species showed specific and distinct tDNA patterns: Streptococcus cristatus, Streptococcus gordonii, Streptococcus oralis, Streptococcus mitis, Streptococcus pneumoniae, Streptococcus sanguinis, Streptococcus parasanguinis, Streptococcus anginosus, Streptococcus mutans, Streptococcus criceti and Streptococcus ratti. Indistinguishable patterns were obtained among two groups of species: Streptococcus vestibularis and Streptococcus salivarius on the one hand and Streptococcus constellatus and Streptococcus intermedius on the other. S. mitis strains produced heterogeneous patterns that could be separated into three groups: a group containing S. mitis biovar 1 and two S. mitis biovar 2 groups, one of which clustered with S. parasanguinis strains while the other showed patterns unrelated to other species. These results agree in part with protein electrophoretic analysis showing that S. mitis biovar 2 strains belong to several streptococcal taxa. In conclusion, PCR analysis of tDNA-ILP holds promise for rapid identification of viridans streptococci that are difficult to identify by phenotypic tests.


Continuity of pediatric ambulatory care in a universally insured population.
To describe the prevalence of continuity of care over a 5-year period in a complete cohort of urban children universally insured for medical care provided under fee-for-service reimbursement. All children enrolled in the study were born to women living in metropolitan Winnipeg between July 1, 1987 and December 31, 1988 (N = 12,590). All ambulatory physician services for this group were enumerated from computerized administrative databases for the period from birth through 60 months. Continuity of care, defined as the proportion of total care provided by the most frequently seen physician or physician practice over time, was calculated for each child. Descriptive analyses include an examination of maternal and household characteristics associated with children receiving 80% or more of total ambulatory care from a single provider source. From birth to 24 months, 51% of children received at least 80% of ambulatory visits from a single provider practice. This proportion of the children declined to 28% at 25 through 60 months. Children living in low-income neighborhoods had poorer continuity profiles. Other household factors associated with poor continuity included young maternal age, single maternal marital status, residential mobility, and inadequate maternal use of prenatal medical care. Households affiliated with pediatric practices had better continuity profiles than households affiliated with general medical practices. Despite universal medical insurance, barriers to a longitudinally continuous relationship with a primary care provider remain in this setting. Although this study has emphasized the description of those barriers associated with household characteristics, there is evidence that factors related to the organization and delivery of medical care are also relevant.