Spiral density waves in a young protoplanetary disk.
Gravitational forces are expected to excite spiral density waves in protoplanetary disks, disks of gas and dust orbiting young stars. However, previous observations that showed spiral structure were not able to probe disk midplanes, where most of the mass is concentrated and where planet formation takes place. Using the Atacama Large Millimeter/submillimeter Array, we detected a pair of trailing symmetric spiral arms in the protoplanetary disk surrounding the young star Elias 2-27. The arms extend to the disk outer regions and can be traced down to the midplane. These millimeter-wave observations also reveal an emission gap closer to the star than the spiral arms. We argue that the observed spirals trace shocks of spiral density waves in the midplane of this young disk.


Mitogens and protein synthesis inhibitors induce ornithine decarboxylase gene transcription through separate mechanisms in the BC3H1 muscle cell line.
Ornithine decarboxylase (ODCase), the rate-limiting enzyme in polyamine biosynthesis, exhibits dramatic fluctuations in activity in response to a variety of hormones and growth factors and has been shown to be down-regulated during myogenesis. In the present study, the molecular mechanisms involved in expression of ODCase mRNA were examined in cells of the BC3H1 muscle line. Proliferating, undifferentiated cells in medium with 20% fetal calf serum displayed high levels of ODCase mRNA and enzyme activity. The transfer of proliferating cells to medium containing 0.5% serum resulted in their withdrawal from the cell cycle and a 20- to 50-fold reduction in the steady-state level of ODCase mRNA within 24 h. Down-regulation of ODCase mRNA was paralleled by a decrease in ODCase enzyme activity and ODCase gene transcription. ODCase mRNA was rapidly reinduced by exposure of quiescent, differentiated cells to medium with 20% serum or by inhibition of protein synthesis with cycloheximide. The accumulation of ODCase mRNA after mitogenic stimulation or protein synthesis inhibition was accompanied by an increase in ODCase gene transcription. The mechanisms whereby mitogens and protein synthesis inhibitors induced ODCase transcription appeared to be different since cycloheximide potentiated the effects of mitogens, resulting in superinduction of ODCase transcription to a level significantly greater than in the presence of mitogens alone. These results indicate that ODCase down-regulation during myogenesis is controlled primarily at the level of ODCase gene transcription. These data also demonstrate that ODCase expression is regulated by antagonistic signals, positive signals for transcription elicited by mitogens and negative signals from endogenous protein repressors that influence ODCase transcription.


Subunit arrangement in the dodecameric chloroplast small heat shock protein Hsp21.
Unfolding proteins are prevented from irreversible aggregation by small heat shock proteins (sHsps) through interactions that depend on a dynamic equilibrium between sHsp subunits and sHsp oligomers. A chloroplast-localized sHsp, Hsp21, provides protection to client proteins to increase plant stress resistance. Structural information is lacking concerning the oligomeric conformation of this sHsp. We here present a structure model of Arabidopsis thaliana Hsp21, obtained by homology modeling, single-particle electron microscopy, and lysine-specific chemical crosslinking. The model shows that the Hsp21 subunits are arranged in two hexameric discs, similar to a cytosolic plant sHsp homolog that has been structurally determined after crystallization. However, the two hexameric discs of Hsp21 are rotated by 25° in relation to each other, suggesting a role for global dynamics in dodecamer function.


Exercise in the fight against thrombosis: friend or foe?
Thrombosis is a major cause of morbidity and mortality in Western countries and is associated with a range of chronic diseases such as cardiovascular disease, renal disease, diabetes, and various autoimmune conditions. Improved health care and approaches to the treatment of disease are leading to aging populations that will probably result in an increase in the incidence of thrombosis and associated manifestations over the next few decades. Adopting a physically active lifestyle through regular exercise has been proposed to lower the risk of developing thrombosis. Indeed, it has been demonstrated that exercise is beneficial for health, although there is inconsistent data from studies investigating the effect of exercise on the risk of thrombosis, with reports of both increased and decreased risk across a variety of cohorts. Studies in this area are difficult to critique due to the variety of confounders such as age, body composition, fitness level, underlying disease and treatment, as well as exercise intensity, frequency, duration, and energy expenditure. In younger individuals and those with chronic conditions such as cardiovascular and kidney disease, there is evidence that physical activity is beneficial for the lowering of thrombotic risk, whereas in older individuals the risk is more likely to be unchanged or increased. This review will explore whether exercise is a "friend" or "foe" in lowering the risk of thrombosis. It will also discuss whether elite athletes have a lower risk of thrombosis and whether exercise may help to reduce thrombotic risk in individuals with chronic disease.


Validity test of the IPD-Work consortium approach for creating comparable job strain groups between Job Content Questionnaire and Demand-Control Questionnaire.
This study aims to test the validity of the IPD-Work Consortium approach for creating comparable job strain groups between the Job Content Questionnaire (JCQ) and the Demand-Control Questionnaire (DCQ). A random population sample (N = 682) of all middle-aged Malmö males and females was given a questionnaire with the 14-item JCQ and 11-item DCQ for the job control and job demands. The JCQ job control and job demands scores were calculated in 3 different ways: using the 14-item JCQ standard scale formulas (method 1); dropping 3 job control items and using the 11-item JCQ standard scale formulas with additional scale weights (method 2); and the approach of the IPD Group (method 3), dropping 3 job control items, but using the simple 11-item summation-based scale formulas. The high job strain was defined as a combination of high demands and low control. Between the 2 questionnaires, false negatives for the high job strain were much greater than false positives (37-49% vs. 7-13%). When the method 3 was applied, the sensitivity of the JCQ for the high job strain against the DCQ was lowest (0.51 vs. 0.60-0.63 when the methods 1 and 2 were applied), although the specificity was highest (0.93 vs. 0.87-0.89 when the methods 1 and 2 were applied). The prevalence of the high job strain with the JCQ (the method 3 was applied) was considerably lower (4-7%) than with the JCQ (the methods 1 and 2 were applied) and the DCQ. The number of congruent cases for the high job strain between the 2 questionnaires was smallest when the method 3 was applied. The IPD-Work Consortium approach showed 2 major weaknesses to be used for epidemiological studies on the high job strain and health outcomes as compared to the standard JCQ methods: the greater misclassification of the high job strain and lower prevalence of the high job strain.


The mayo classification system for traumatic brain injury severity.
To develop a single TBI severity classification system based on commonly used TBI severity measures and indicators that (1) maximally uses available positive evidence to classify TBI severity in three categories: (a) Moderate-Severe (Definite) TBI, (b) Mild (Probable) TBI, (c) Symptomatic (Possible) TBI; (2) reflects current clinical knowledge and relevance; and (3) classifies a larger number of cases than single indicator systems with reasonable accuracy. The study sample of a defined population consisted of 1501 unique Olmsted County residents with at least one confirmed TBI event from 1985 to 1999. Within the sample, 1678 TBI events were confirmed. Single measures of TBI severity were not available in a large percentage of these events, i.e., Glasgow Coma Scale (GCS) was absent in 1242 (74.0%); loss of consciousness, absent in 178 (70.2%), posttraumatic amnesia (PTA), absent in 974 (58.1%), head CT, not done in 827 (49.3%). The Mayo Classification System for TBI Severity was developed to classify cases based on available indicators that included death due to TBI, trauma-related neuroimaging abnormalities, GCS, PTA, loss of consciousness and specified post-concussive symptoms. Using the Mayo system, all cases were classified. For the Moderate-Severe (Definite) TBI classification, estimated sensitivity was 89% and estimated specificity was 98%. By maximally using relevant available positive evidence, the Mayo system classifies a larger number of cases than single indicator systems with reasonable accuracy. This system may be of use in retrospective research and for determination of TBI severity for planning postacute clinical care.


Heterotopic hairs in the tongue of two dogs.
The occurrence of hairs in the median sulcus of the tongue in two dogs, a 7-year-old Rottweiler and a 10-year-old Cocker spaniel, is reported as an incidental observation at necropsy. The gross pathology of both cases and the histological and scanning electron microscopical findings in one case indicated a heterotopic development of the hairs, with secondary inflammation. The term pili heterotopici mediani linguae is suggested.


Process and outcome constructs for evaluating community-based participatory research projects: a matrix of existing measures.
Community-based participatory research (CBPR) has been widely used in public health research in the last decade as an approach to develop culturally centered interventions and collaborative research processes in which communities are directly involved in the construction and implementation of these interventions and in other application of findings. Little is known, however, about CBPR pathways of change and how these academic-community collaborations may contribute to successful outcomes. A new health CBPR conceptual model (Wallerstein N, Oetzel JG, Duran B et al. CBPR: What predicts outcomes? In: Minkler M, Wallerstein N (eds). Communication Based Participatory Research, 2nd edn. San Francisco, CA: John Wiley & Co., 2008) suggests that relationships between four components: context, group dynamics, the extent of community-centeredness in intervention and/or research design and the impact of these participatory processes on CBPR system change and health outcomes. This article seeks to identify instruments and measures in a comprehensive literature review that relates to these distinct components of the CBPR model and to present them in an organized and indexed format for researcher use. Specifically, 258 articles were identified in a review of CBPR (and related) literature from 2002 to 2008. Based on this review and from recommendations of a national advisory board, 46 CBPR instruments were identified and each was reviewed and coded using the CBPR logic model. The 46 instruments yielded 224 individual measures of characteristics in the CBPR model. While this study does not investigate the quality of the instruments, it does provide information about reliability and validity for specific measures. Group dynamics proved to have the largest number of identified measures, while context and CBPR system and health outcomes had the least. Consistent with other summaries of instruments, such as Granner and Sharpe's inventory (Granner ML, Sharpe PA. Evaluating community coalition characteristics and functioning: a summary of measurement tools. Health Educ Res 2004; 19: 514-32), validity and reliability information were often lacking, and one or both were only available for 65 of the 224 measures. This summary of measures provides a place to start for new and continuing partnerships seeking to evaluate their progress.


Distribution of Barley yellow dwarf virus-PAV in the Sub-Antarctic Kerguelen Islands and Characterization of Two New Luteovirus Species.
A systematic search for viral infection was performed in the isolated Kerguelen Islands, using a range of polyvalent genus-specific PCR assays. Barley yellow dwarf virus (BYDV) was detected in both introduced and native grasses such as Poa cookii. The geographical distribution of BYDV and its prevalence in P. cookii were analyzed using samples collected from various sites of the archipelago. We estimate the average prevalence of BYDV to be 24.9% in P. cookii, with significant variability between sites. BYDV genetic diversity was assessed using sequence information from two genomic regions: the P3 open reading frame (ORF) (encoding the coat protein) and the hypervariable P6 ORF region. The phylogenetic analysis in the P3 region showed that BYDV sequences segregate into three major lineages, the most frequent of which (Ker-I cluster) showed close homology with BYDV-PAV-I isolates and had very low intra-lineage diversity (0.6%). A similarly low diversity was also recorded in the hypervariable P6 region, suggesting that Ker-I isolates derive from the recent introduction of BYDV-PAV-I. Divergence time estimation suggests that BYDV-PAV-I was likely introduced in the Kerguelen environment at the same time frame as its aphid vector, Rhopalosiphum padi, whose distribution shows good overlap with that of BYDV-Ker-I. The two other lineages show more than 22% amino acid divergence in the P3 region with other known species in the BYDV species complex, indicating that they represent distinct BYDV species. Using species-specific amplification primers, the distribution of these novel species was analyzed. The high prevalence of BYDV on native Poaceae and the presence of the vector R. padi, raises the question of its impact on the vulnerable plant communities of this remote ecosystem.


Kinetics of O2 Entry and Exit in Monomeric Sarcosine Oxidase via Markovian Milestoning Molecular Dynamics.
The flavoenzyme monomeric sarcosine oxidase (MSOX) catalyzes a complex set of reactions currently lacking a consensus mechanism. A key question that arises in weighing competing mechanistic models of MSOX function is to what extent ingress of O2 from the solvent (and its egress after an unsuccessful oxidation attempt) limits the overall catalytic rate. To address this question, we have applied to the MSOX/O2 system the relatively new simulation method of Markovian milestoning molecular dynamics simulations, which, as we recently showed [ Yu et al. J. Am. Chem. Soc. 2015 , 137 , 3041 ], accurately predicted the entry and exit kinetics of CO in myoglobin. We show that the mechanism of O2 entry and exit, in terms of which possible solvent-to-active-site channels contribute to the flow of O2, is sensitive to the presence of the substrate-mimicking competitive inhibitor 2-furoate in the substrate site. The second-order O2 entry rate constants were computed to be 8.1 × 10(6) and 3.1 × 10(6) M(-1) s(-1) for bound and apo MSOX, respectively, both of which moderately exceed the experimentally determined second-order rate constant of (2.83 ± 0.07) × 10(5) M(-1) s(-1) for flavin oxidation by O2 in MSOX. This suggests that the rate of flavin oxidation by O2 is likely not strongly limited by diffusion from the solvent to the active site. The first-order exit rate constants were computed to be 10(7) s(-1) and 7.2 × 10(6) s(-1) for the apo and bound states, respectively. The predicted faster entry and slower exit of O2 for the bound state indicate a longer residence time within MSOX, increasing the likelihood of collisions with the flavin isoalloxazine ring, a step required for reduction of molecular O2 and subsequent reoxidation of the flavin. This is also indirectly supported by previous experimental evidence favoring the so-called modified ping-pong mechanism, the distinguishing feature of which is an intermediate complex involving O2, the flavin, and the oxidized substrate simultaneously in the cavity. These findings demonstrate the utility of the Markovian milestoning approach in contributing new understanding of complicated enyzmatic function.


Evaluation of the minimum infectious dose of porcine epidemic diarrhea virus in virus-inoculated feed.
OBJECTIVE To determine the minimum infectious dose of porcine epidemic diarrhea virus (PEDV) in virus-inoculated feed. ANIMALS 30 crossbred 10-day-old pigs. PROCEDURES Tissue culture PEDV was diluted to form 8 serial 10-fold dilutions. An aliquot of stock virus (5.6 × 10(5) TCID50/mL) and each serial PEDV dilution were mixed into 4.5-kg batches of feed to create 9 PEDV-inoculated feed doses; 1 virus-negative dose of culture medium in feed was also created. Pigs were challenge exposed via oral administration of PEDV-inoculated feed, and fecal swab specimens were collected. All pigs were euthanized 7 days after challenge exposure; fresh tissues were collected and used for PCR assay, histologic examination, and immunohistochemical analysis. RESULTS The PCR cycle threshold (Ct) decreased by approximately 10 when PEDV was added to feed, compared with results for equivalent PEDV diluted in tissue culture medium. Pigs became infected with PEDV when challenge exposed with the 4 highest concentrations (lowest concentration to cause infection, 5.6 × 10(1) TCID50/g; Ct = 27 in tissue culture medium and 37 in feed). CONCLUSIONS AND CLINICAL RELEVANCE In this study, PEDV in feed with detectable Ct values of 27 to 37 was infective. The Ct was 37 for the lowest infective PEDV dose in feed, which may be above the limit of detection established for PEDV PCR assays used by some diagnostic laboratories. Overall, results indicated 5.6 × 10(1) TCID50/g was the minimum PEDV dose in feed that can lead to infection in 10-day-old pigs under the conditions of this study.


Domains of increased thickness in microvillar membranes of the small intestinal enterocyte.
The apical surface of the enterocyte is sculpted into a dense array of cylindrical microvillar protrusions by supporting actin filaments. Membrane microdomains (rafts) enriched in cholesterol and glycosphingolipids comprise roughly 50% of the microvillar membrane and play a vital role in orchestrating absorptive/digestive action of dietary nutrients at this important cellular interface. Increased membrane thickness is believed to be a morphological characteristic of rafts. Thus, we investigated whether the high contents of lipid rafts in the microvillar membrane is reflected in local variations in membrane thickness. We measured membrane thickness directly from electron micrographs of sections of fixed mucosal tissue. Indeed, mapping of the microvillar membrane revealed a biphasic distribution of membrane thickness. As a point of reference the thickness distribution of the basolateral membrane was clearly monophasic. The encountered domains of increased thickness (DITs) occupied 48% of the microvillar membrane and from the data we estimated the area of a single DIT to have a lower limit of 600 nm(2). In other experiments we mapped the organization of biochemically defined lipid rafts by immunogold labeling of alkaline phosphatase, a well documented raft marker. Strikingly, the alkaline phosphatase localized to distinct regions of the membrane in a pattern similar to the observed distribution of DITs. Although we were unable to measure membrane thickness directly on the immunogold labeled specimens, and thereby establish an unequivocal connection between DITs and rafts, we conclude that the brush border membrane of the enterocyte contains microdomains distinguishable both by membrane morphology and protein composition.


Enhanced desensitization and phosphorylation of the beta 1-adrenergic receptor in rat adipocytes by peroxovanadate.
Peroxovanadate (PVN) is an insulin-like agent that inhibits the dephosphorylation of the insulin receptor kinase. PVN inhibited the lipolytic action of 0.1 microM isoproterenol by 88%, which is a relatively specific beta 1 catecholamine agonist at this concentration, but was largely ineffective against beta 3 agonists or forskolin. To determine whether PVN-mediated desensitization of the beta 1 AR was associated with enhanced phosphorylation, we immunoprecipitated the beta 1 AR from rat adipocytes that were metabolically labeled with 32PO4. Isoproterenol enhanced the net phosphorylation of the beta 1 AR by 8 +/- 2-fold over control. PVN increased the net phosphorylation of the beta 1 AR by 5 +/- 0.5-fold, and together with isoproterenol, they enhanced the phosphorylation of the beta 1 AR by 2-fold over isoproterenol alone. Phosphoamino acid analysis of the phosphorylated receptor revealed phosphate incorporation into serine that was proportional to the radioactivity incorporated into the immunoprecipitated receptor. PVN inhibited the serine/threonine phosphatase calcineurin, suggesting that inhibition of receptor dephosphorylation may play a role in the actions of PVN. Cyanogen bromide cleavage of the phosphorylated beta 1 AR generated a phosphoprotein with a molecular mass consistent with carboxyl-terminal phosphorylation. Furthermore, the magnitude of receptor phosphorylation by isoproterenol was 3-fold larger than that due to forskolin, suggesting that beta 1 AR is a substrate for the beta AR kinase that phosphorylates carboxyl-terminal residues in the beta(2) AR. Our findings suggest that PVN may be a powerful new tool with which to study the phosphorylation of other G protein-coupled receptors.


On the possibility of quantitative refractive-index tomography of large biomedical samples with hard X-rays.
One of the most promising applications of the X-ray phase-contrast imaging is the three dimensional tomographic reconstruction of the index of refraction. However, results reported so far are limited to relatively small samples. We present here the tomographic reconstruction of the index of refraction distribution of a large biomedical sample (> 10 cm diameter). A quantitative study comparing the absorption and phase contrast (analyzer-based) tomography images shows that the distribution of the index of refraction obtained with the phase contrast method provides a more accurate depiction (3-10 times larger signal to noise ratio values) of the sample internal structure. Thanks to the higher sensitivity of this method, the improved precision was obtained using an incoming photon fluence on the sample several times smaller than in the case of absorption imaging.


Mirex-induced impairment of hepatobiliary function. Suppressed biliary excretion of imipramine and sulfobromophthalein.
Effect of pre-exposure to the pesticide mirex on hepatic uptake, metabolism, and biliary excretion of the tricyclic antidepressant, imipramine (IMP), was studied by using isolated perfused rat liver preparations obtained from control and treated animals. Pretreatment of male rats with mirex (50 mg/kg/day, po, for 3 days) resulted in a marked suppression (91%) of biliary excretion of endogenously formed polar metabolites of IMP. This impairment of biliary excretory function is related neither to the bile flow nor to the rate of metabolism of IMP. These conclusions are supported by the following observations: 1) pre-exposure to mirex results in a significant increase in bile secretion; 2) endogenously formed metabolites of IMP accumulate in increasing concentrations in the perfusate of liver preparations from mirex-treated rats; 3) over 98% of the IMP was metabolized by livers of both control and mirex-treated rats. Furthermore, biliary excretion of exogenously provided polar metabolites of IMP was also suppressed by 88% of that of control livers. Addition of mirex (5 X 10(-5) M) to the perfusate of control liver preparations was without effect on biliary excretion of IMP metabolites. Biliary excretion of sulfobromophthalein was also markedly suppressed (90%) by preexposure to mirex. These results suggest that mirex-induced impairment of hepatobiliary function is not specific to substrates. The mechanism of mirex-induced impairment is located at the site of transfer of otherwise readily excretable substances such as metabolites of IMP and sulfobromophthalein.


Are Improvements Still Needed to the Modified Hospital Readmissions Reduction Program: a Health and Retirement Study (2000-2014)?
To address concerns that the Hospital Readmissions Reduction Program (HRRP) unfairly penalized safety net hospitals treating patients with high social and functional risks, Medicare recently modified HRRP to compare hospitals with similar proportions of high-risk, dual-eligible patients ("peer group hospitals"). Whether the change fully accounts for patients' social and functional risks is unknown. Examine risk-standardized readmission rates (RSRRs) and hospital penalties after adding patient-level social and functional and community-level risk factors. Using 2000-2014 Medicare hospital discharge, Health and Retirement Study, and community-level data, latent factors for patient social and functional factors and community factors were identified. We estimated RSRRs for peer groups and by safety net status using four hierarchical logistic regression models: "base" (HRRP model); "patient" (base plus patient factors); "community" (base plus community factors); and "full" (all factors). The proportion of hospitals penalized was calculated by safety net status. 20,255 fee-for-service Medicare beneficiaries (65+) with eligible index hospitalizations MAIN MEASURES: RSRRs KEY RESULTS: Half of safety net hospitals are in peer group 5. Compared with other hospitals, peer group 5 hospitals (most dual-eligibles) treated sicker, more functionally limited patients from socially disadvantaged groups. RSRRs decreased by 0.7% for peer groups 2 and 4 and 1.3% for peer group 5 under the patient and full (versus base) models. Measured performance improved after adjusting for patient risk factors for hospitals in peer group 4 and 5 hospitals, but worsened for those in peer groups 1, 2, and 3. Under the patient (versus base) model, fewer safety net hospitals (48.7% versus 51.3%) but more non-safety net hospitals (50.0% versus 49.1%) were penalized. Patient-level risk adjustment decreased RSRRs for hospitals serving more at-risk patients and proportion of safety net hospitals penalized, while modestly increasing RSRRs and proportion of non-safety net hospitals penalized. Results suggest HRRP modifications may not fully account for hospital variation in patient-level risk.


Evaluation of doctoral nursing programs in Japan by faculty members and their educational and research activities.
Evaluation of doctoral programs in nursing is becoming more important with the rapid increase in the programs in Japan. This study aimed to evaluate doctoral nursing programs by faculty members and to analyze the relationship of the evaluation with educational and research activities of faculty members in Japan. Target settings were all 46 doctoral nursing programs. Eighty-five faculty members from 28 programs answered the questionnaire, which included 17 items for program evaluation, 12 items for faculty evaluation, 9 items for resource evaluation, 3 items for overall evaluations, and educational and research activities. A majority gave low evaluations for sources of funding, the number of faculty members and support staff, and administrative systems. Faculty members who financially supported a greater number of students gave a higher evaluation for extramural funding support, publication, provision of diverse learning experiences, time of supervision, and research infrastructure. The more time a faculty member spent on advising doctoral students, the higher were their evaluations on the supportive learning environment, administrative systems, time of supervision, and timely feedback on students' research. The findings of this study indicate a need for improvement in research infrastructure, funding sources, and human resources to achieve quality nursing doctoral education in Japan.


Organization of hindlimb muscle afferent projections to lumbosacral motoneurons in the chick embryo.
We have examined the organization of muscle afferent projections to motoneurons in the lumbosacral spinal cord of chick embryos between stage 37, when muscle afferents first reach the motor nucleus, and stage 44, which is just before hatching. Connectivity between afferents and motoneurons was assessed by stimulating individual muscle nerves and recording the resulting motoneuron synaptic potentials intracellularly or electrotonically from other muscle nerves. Most of the recordings were made in the presence of DL-2-amino-5-phosphonovaleric acid (APV), picrotoxin, and strychnine to block long-latency excitatory and inhibitory pathways. Activation of muscle afferents evoked slow, positive potentials in muscle nerves but not in cutaneous nerves. These potentials were abolished in 0 mM Ca2+, 2mM Mn2+ solutions, indicating that they were generated by the action of chemical synapses. The muscle nerve recordings revealed a wide-spread pattern of excitatory connections between afferents and motoneurons innervating six different thigh muscles, which were not organized according to synergist-antagonist relationships. This pattern of connectivity was confirmed using intracellular recording from identified motoneurons, which allowed the latency of the responses to be determined. Short-latency potentials in motoneurons were produced by activation of homonymous afferents and the heteronymous afferents innervating the hip flexors sartorius and anterior iliotibialis. Stimulation of anterior iliotibialis afferents also resulted in some short-latency excitatory postsynaptic potentials (EPSPs) in motoneurons innervating the knee extensor femorotibialis, though other connections were of longer latency. Afferents from the adductor, a hip extensor, did not evoke short-latency EPSPs in any of these three types of motoneurons. Short-latency, but not long-latency EPSPs, persisted during repetitive stimulation at 5 Hz, suggesting that they were mediated monosynaptically. Long-latency, fatigue-sensitive potentials were maintained in the presence of APV, picrotoxin, and strychnine, suggesting that polysynaptic pathways utilize non-NMDA receptors as well as NMDA receptors. We found no difference in the pattern of inputs to femorotibialis motoneurons between stage 37-39 and near hatching at stage 44, suggesting muscle afferent projections to these motoneurons are correct at stage 37, when the afferents first reach the lateral motor column in substantial numbers.


Long-lived charge-separated state generated in a ferrocene-meso,meso-linked porphyrin trimer-fullerene pentad with a high quantum yield.
A meso,meso-linked porphyrin trimer, (ZnP)3, as a light-harvesting chromophore, has been incorporated for the first time into a photosynthetic multistep electron-transfer model including ferrocene (Fc) as an electron donor and fullerene (C60) as an electron acceptor, to construct the ferrocene-meso,meso-linked porphyrin trimer-fullerene system Fc-(ZnP)3-C60. Photoirradiation of Fc-(ZnP)3-C60 results in photoinduced electron transfer from both the singlet and triplet excited states of the porphyrin trimer, 1(ZnP)3* and 3(ZnP)3*, to the C60 moiety to produce the porphyrin trimer radical cation-C60 radical anion pair, Fc-(ZnP)3*+-C60*-. Subsequent formation of the final charge-separated state Fc+-(ZnP)3-C60*- was confirmed by the transient absorption spectra observed by pico- and nanosecond time-resolved laser flash photolysis. The final charge-separated state decays, obeying first-order kinetics, with a long lifetime (0.53 s in DMF at 163 K) that is comparable with that of the natural bacterial photosynthetic reaction center. More importantly, the quantum yield of formation of the final charge-separated state (0.83 in benzonitrile) remains high, despite the large separation distance between the Fc+ and C60*- moieties. Such a high quantum yield results from efficient charge separation through the porphyrin trimer, whereas a slow charge recombination is associated with the localized porphyrin radical cation in the porphyrin trimer. The light-harvesting efficiency in the visible region has also been much improved in Fc-(ZnP)3-C60 because of exciton coupling in the porphyrin trimer as well as an increase in the number of porphyrins.


Evaluation of a commercial radioisotopic kit for folate assays.
A commercial kit for the radioisotopic assay of folate in serum, the Bio-Rad 'Quanta Count' folate kit, produced lower results than the Lactobacillus casei microbiological assay method. Its normal range was 2-0-13-0 microng/l and the reproducibility was similar to that of the microbiological assay method. The kit was also satisfactory for whole bood folate assays. The cost requires careful consideration before the kit is used for routine purposes.


A simple high-yield synthesis of high-purity Hägg carbide (χ-Fe5C2) nanoparticles with extraordinary electrochemical properties.
Iron carbides are of eminent interest in both fundamental scientific research and in the industry owing to their properties such as excellent mechanical strength and chemical inertness. They have been found very effective in Fischer-Tropsch synthesis exploring heterogeneous catalysis for the production of chemicals such as liquid fuel and they have also been employed as successful promoters for the oxygen reduction reaction (ORR) and hydrogen evolution reaction (HER). However, so far there have been only a few reports on the application of iron carbide nanoparticles in the field of electrochemical sensing. Here, we present a stable form of Hägg carbide nanoparticles synthesized from a rare form of iron(iii) oxide (β-Fe2O3). The as-prepared nanomaterial was characterized employing X-ray powder diffraction and Mössbauer spectroscopy to prove its composition as well as an extraordinary high purity level. It turned out that Hägg carbide nanoparticles prepared by thermally treated β-Fe2O3 exhibited excellent electrochemical properties including low charge transfer resistivity (Rct) compared to the other tested materials. Moreover, the Hägg carbide nanoparticles were tested as a promising electrocatalyst for voltammetric detection of the antibiotic metronidazole proving its practical applicability.


Activation of human eosinophils by IL-13. Induction of CD69 surface antigen, its relationship to messenger RNA expression, and promotion of cellular viability.
To study the effect of IL-13 on CD69 expression and cell viability in human eosinophils, purified human peripheral blood eosinophils from healthy donors were incubated with increasing concentrations of IL-13. The expression of CD69 was analyzed by flow cytometry (FACS). Surface expression of CD69, which was absent on untreated eosinophils, was induced by IL-13 at concentrations ranging from 1 ng/ml to 1 microgram/ml in a concentration-dependent manner. In contrast, neutrophils expressed CD69 neither spontaneously nor following incubation with IL-13. Semiquantitative reverse transcription-PCR for CD69 mRNA showed constitutive CD69 mRNA expression in purified human eosinophils. Incubation of eosinophils with IL-13 further increased CD69-specific mRNA. Analysis of intracellularly stored CD69 in eosinophils permeabilized with saponin revealed intracellular binding of anti-CD69 Abs in all isolated eosinophils. After stimulation with 100 ng/ml IL-13 for 24 h, the concentration of intracellular CD69 decreased by 41 +/- 9%. Furthermore, IL-13 at a concentration of 100 ng/ml enhanced eosinophil viability, as assessed by propidium iodide staining after 4 days in culture from 8.6 +/- 5.5% in control medium to 50.7 +/- 6.8% following stimulation with IL-13 treatment. The effect of Il-13 on eosinophil viability as well as that on CD69 expression were both neutralized by anti-IL-13 Abs. In conclusion, our results demonstrate that IL-13 specifically activates human eosinophils, as determined by the expression of CD69 cell surface protein and mRNA expression. Furthermore, IL-13 significantly prolongs eosinophil survival in vitro. The data suggest that IL-13 may play a role in the activation of eosinophils.


Properties of multiple G.A mismatches in stable oligonucleotide duplexes.
The solution structure of the deoxydecanucleotide [sequence: see text] has been determined by NMR methods. This duplex, which contains six G.A mismatches and four Watson-Crick base pairs, is thermodynamically more stable than a decamer where T.A base pairs are substituted for the G.A mismatches, and is less stable than the duplex that contains G.C base pairs. Circular-dichroism spectroscopy indicates an overall B-like conformation for the decamer, but stronger than usual base stacking. 1H-NMR spectroscopy revealed that the N1H groups of the mismatched guanine residues are not hydrogen bonded, and 31P-NMR showed the presence of BII phosphate conformations for the GpA steps. Detailed analysis of the NMR data showed that all nucleotides have anti glycosidic torsion angles and S type sugar puckers. The G.A mismatches pair in the amino form as originally proposed by Li et al. [Li, Y., Zon, G. & Wilson, W. D. (1991) Proc. Natl Acad. Sci. USA 88, 26-30], which results in extensive base-base stacking between the tandem G.A base pairs and their nearest neighbours. The terminal G.A base pairs are less stable than the central base pairs and show evidence of an equilibrium between two conformations, one involving BII phosphate.


Associations between biological markers of prenatal stress and infant negative emotionality are specific to sex.
Fetal programming is the idea that environmental stimuli can alter the development of the fetus, which may have a long-term effect on the child. We have recently reported that maternal prenatal cortisol predicts infant negative emotionality in a sex-dependent manner: high prenatal cortisol was associated with increased negative emotionality in females, and decreased negative emotionality in males. This study aims to test for this sex-specific effect in a different cohort, and investigate whether sex differences in fetal programming may be specific to glucocorticoid mechanisms by also examining a maternal salivary alpha-amylase (sAA) by sex interaction. 88 pregnant women (mean gestational age=27.4 weeks, SD=7.4) collected saliva samples at home over two working days to be assayed for the hormone cortisol (range=0.13-88.22nmol/l) and the enzyme alpha-amylase (range=4.57-554.8units/ml). Samples were collected at waking, 30-min post-waking and 12h post-waking. Two months after birth participants reported infant negative emotionality using the distress to limits subscale of the Infant Behavior Questionnaire. The interaction between maternal prenatal cortisol and infant sex to predict distress to limits approached significance (p=0.067). In line with our previous finding there was a positive association between prenatal cortisol and negative emotionality in females, and a negative association in males. The interaction between sAA and sex to predict distress was significant (p=0.025), and the direction of effect was the same as for the cortisol data; high sAA associated with increased negative emotionality in females and reduced negative emotionality in males. In line with our previous findings, this research adds to an emerging body of literature, which suggests that fetal programming mechanisms may be sex-dependent. This is the first study to demonstrate that maternal prenatal sAA may be an important biomarker for infant behavior, and the findings have implications for understanding sex differences in developmental psychopathology.


Molecular Engineering of D-π-A Type of Blue-Colored Dyes for Highly Efficient Solid-State Dye-Sensitized Solar Cells through Co-Sensitization.
A novel blue-colored organic donor-π-acceptor sensitizer, the so-called MKA16 dye, has been employed to construct solid-state dye-sensitized solar cells (ssDSSCs). Using 2,2',7-,7'-tetrakis( N, N-di- p-methoxyphenyl-amine) 9,9'-spirobifuorene (Spiro-OMeTAD) as hole-transport material, a good conversion efficiency of 5.8% was recorded for cells based on the MKA16 dye and a high photovoltage of 840 mV in comparison with 5.6% efficiency using the known (Dyenamo Blue) dye. By co-sensitization using the orange-colored D35 dye and MKA16 together, the solid-state solar cells showed an excellent efficiency of 7.5%, with a high photocurrent of 12.41 mA cm-2 and open-circuit voltage of 850 mV. The results show that the photocurrent of ssDSSCs can be significantly improved by co-sensitization mainly attributed to the wider light absorption range contributing to the photocurrent. In addition, results from photo-induced absorption spectroscopy show that the dye regeneration is efficient in co-sensitized solar cells. The current results possible routes of improving the design of aesthetic and highly efficient ssDSSCs.


Sympathetic improvement of cancer vaccine efficacy.
The link between stress, other psychological factors and response to cancer, or even the cancer incidence and metastasis, is well established. The inhibition of β-Adrenergic receptors (β-AR) using β-blockers was demonstrated to have an inhibitory effect on cancer recurrence. Direct effects on the stress-induced suppression of anti-tumor immune responses were also shown. In a recent issue of Cancer Immunology Research, Daher and colleagues studied the molecular mechanism behind this protective effect in the context of cancer vaccination. They provided evidence that the β-AR signaling affected the priming of naïve CD8 + T cells in their myeloma model, rather than effector CD8 + T cells which downregulated the expression of β-AR after activation and became insensitive to such signaling. Blocking the β-adrenergic signaling during vaccination led to increased expansion and effector functions of antigen-specific CD8 + T cells and reduced tumor growth. This has implications for the clinical use of β-blockers as adjuvants to enhance cancer vaccination and other types of immunotherapy.


Food provision and the nutritional implications of food choices made by young adult males, in a young offenders' institution.
The nutritional adequacy of diets provided by a prison was assessed by analysis of the kitchen menu for 1 week of a 4-week cycle. Dietary intakes were determined using a predefined 7-day diet diary in which prisoners indicated what they had eaten, and how much. A total of 159 prisoners took part in the study. The food provided by the prison kitchen was broadly in line with current dietary recommendations. Vitamin content exceeded recommendations, with the exception of niacin in the vegetarian menu (12.6 mg compared with the reference nutrient intake of 16.8 mg). Selenium content was low in all menus, but particularly in the vegetarian menu in 1997 where it equalled the lower reference nutrient intake (LRNI) (39.5 microg). Food choices made by prisoners resulted in a wide variation in dietary intakes. Fat intake (as a proportion of energy) exceeded the recommended 35% in 82% of diets in 1996, and 64% of diets in 1997. In 1996, 34% of prisoners had intakes above 40% energy as fat. High fat intakes were largely the result of consuming items from the prison shop. Vitamin D intakes were low (3.4 and 3.3 microg in 1996 and 1997, respectively) compared with the recommendation (10 microg) for those with limited exposure to sunlight. Intakes of a number of minerals fell below recommendations, with some prisoners barely meeting the LRNI. This was particularly notable for selenium where 35% of prisoners in 1996, and 60% of prisoners in 1997 had intakes below the LRNI.


Regulation of aging and age-related disease by DAF-16 and heat-shock factor.
The Caenorhabditis elegans transcription factor HSF-1, which regulates the heat-shock response, also influences aging. Reducing hsf-1 activity accelerates tissue aging and shortens life-span, and we show that hsf-1 overexpression extends lifespan. We find that HSF-1, like the transcription factor DAF-16, is required for daf-2-insulin/IGF-1 receptor mutations to extend life-span. Our findings suggest this is because HSF-1 and DAF-16 together activate expression of specific genes, including genes encoding small heat-shock proteins, which in turn promote longevity. The small heat-shock proteins also delay the onset of polyglutamine-expansion protein aggregation, suggesting that these proteins couple the normal aging process to this type of age-related disease.


Induction of the neural crest state: control of stem cell attributes by gene regulatory, post-transcriptional and epigenetic interactions.
Neural crest cells are a population of multipotent stem cell-like progenitors that arise at the neural plate border in vertebrates, migrate extensively, and give rise to diverse derivatives such as melanocytes, craniofacial cartilage and bone, smooth muscle, peripheral and enteric neurons and glia. The neural crest gene regulatory network (NC-GRN) includes a number of key factors that are used reiteratively to control multiple steps in the development of neural crest cells, including the acquisition of stem cell attributes. It is therefore essential to understand the mechanisms that control the distinct functions of such reiteratively used factors in different cellular contexts. The context-dependent control of neural crest specification is achieved through combinatorial interaction with other factors, post-transcriptional and post-translational modifications, and the epigenetic status and chromatin state of target genes. Here we review the current understanding of the NC-GRN, including the role of the neural crest specifiers, their links to the control of "stemness," and their dynamic context-dependent regulation during the formation of neural crest progenitors.


Insight and prefrontal cortex in first-episode Schizophrenia.
Few studies have investigated the neurobiological basis of impaired insight in antipsychotic-naive schizophrenia. However, the relationship between insight and specific prefrontally mediated cognitive functions suggests that insight deficits may be an expression of prefrontal cortical dysfunction. This study was designed to examine the relationship among insight, neurocognition, and dorsolateral prefrontal cortex (DLPFC) volumes in first-episode antipsychotic-naive schizophrenia subjects. DLPFC volumes were compared between 35 first-episode schizophrenia subjects with good (n = 17) and poor insight (n = 18). Morphometric measurements were based on MRI scans by trained raters blind to clinical information. First-episode schizophrenia subjects with poor insight showed decreased right DLPFC volumes relative to those with good insight. In addition, those with poor insight had higher levels of perseverative errors (PEs) on the Wisconsin Card Sort Test (WCST). No differences in other neuropsychological measures were found between the good and poor insight groups. Similarly, no differences were found between schizophrenia subjects with good versus poor insight on any of the psychopathological measures employed in this study. These findings suggest that poor insight in schizophrenia may be a function of specific prefrontally mediated neurocognitive deficits rather than a global impairment in neuropsychological functioning or different profiles of psychopathology.


Influence of non-Newtonian behavior of blood on flow in an elastic artery model.
Two different non-Newtonian models for blood, one a simple power law model exhibiting shear thinning viscosity, and another a generalized Maxwell model displaying both shear thinning viscosity and oscillatory flow viscoelasticity, were used along with a Newtonian model to simulate sinusoidal flow of blood in rigid and elastic straight arteries. When the spring elements were removed from the viscoelastic model resulting in a purely viscous shear thinning fluid, the predictions of flow rate and WSS were virtually unaltered. Hence, elasticity of blood does not appear to influence its flow behavior under physiological conditions in large arteries, and a purely viscous shear thinning model should be quite realistic for simulating blood flow under these conditions. When a power law model with a high shear rate Newtonian cutoff was used for sinusoidal flow simulation in elastic arteries, the mean and amplitude of the flow rate were found to be lower for a power law fluid compared to a Newtonian fluid experiencing the same pressure gradient. The wall shear stress was found to be relatively insensitive to fluid rheology but strongly dependent on vessel wall motion for flows driven by the same pressure gradient. The effect of wall motion on wall shear stress could be greatly reduced by matching flow rate rather than pressure gradient. For physiological flow simulation in the aorta, an increase in mean WSS but a reduction in peak WSS were observed for the power law model compared to a Newtonian fluid model for a matched flow rate waveform.


Physiological, biochemical and genetic responses of Caucasian tea (Camellia sinensis (L.) Kuntze) genotypes under cold and frost stress.
Cold and frost are two serious factors limiting the yield of many crops worldwide, including the tea plant (Camellia sinensis (L.) Kuntze). The acclimatization of tea plant from tropical to temperate climate regions resulted in unique germplasm in the North-Western Caucasus with extremely frost-tolerant genotypes. The aim of the current research was to evaluate the physiological, biochemical and genetic responses of tolerant and sensitive tea cultivars exposed to cold (0 to +2 °C for 7 days) and frost (-6 to -8 °C for 5 days). Relative water content, cell membranes integrity, pH of the cell sap, water soluble protein, cations, sugars, amino acids were measured under cold and frost. Comparative expression of the following genes ICE1, CBF1, WRKY2, DHN1, DHN2, DHN3, NAC17, NAC26, NAC30, SnRK1.1, SnRK1.2, SnRK1.3, bHLH7, bHLH43, P5CS, LOX1, LOX6, LOX7 were analyzed. We found elevated protein (by 3-4 times) and cations (potassium, calcium and magnesium) contents in the leaves of both cultivars under cold and frost treatments. Meanwhile, Leu, Met, Val, Thr, Ser were increased under cold and frost, however tolerant cv. Gruzinskii7 showed earlier accumulation of these amino acids. Out of 18 studied genes, 11 were expressed at greater level in the frost- tolerant cultivar comparing with frost-sensitive one: ICE1, CBF1, WRKY2, DHN2, NAC17, NAC26, SnRK1.1, SnRK1.3, bHLH43, P5CS and LOX6. Positive correlations between certain amino acids namely, Met, Thr, Leu and Ser and studied genes were found. Taken together, the revealed cold responses in Caucasian tea cultivars help better understanding of tea tolerance to low temperature stress and role of revealed metabolites need to be further evaluated in different tea genotypes.


The effect of statin therapy on testosterone levels in subjects consulting for erectile dysfunction.
Previous clinical studies on effect of statins treatment on testosterone (T) levels have produced mixed results. The aim of the present study is to evaluate the association between statin therapy and hormonal parameters in a large series of subjects seeking medical care at our unit for erectile dysfunction (ED). A consecutive series of 3,484 (mean age 51.6 + or - 13.1 years) patients with ED was studied. Several hormonal and biochemical parameters were investigated, along with ANDROTEST structured interview measuring hypogonadism-related symptoms. Among the patients studied, 244 (7%) patients were being treated with statins. After adjustment for confounding factors (including body mass index and Progetto Cuore cardiovascular (CV) risk engine score), both total and calculated free testosterone levels were significantly lower in subjects taking statins, when compared to the rest of the sample (hazard ratio [HR] = 0.93 [0.90; 0.96] and 0.26 [0.01; 0.18] for each decrement of total T and calculated free T, respectively; both P < 0.0001). The use of statins was also associated with a reduced testis volume and a higher prevalence of hypogonadism-related symptoms and signs, as assessed by higher ANDROTEST score (HR = 1.12 [1.03; 1.21]; P < 0.01 after adjustment for confounders). Follicle-stimulating hormone levels were significantly higher in subjects treated with statins when compared to the rest of the sample, while there was a trend toward higher luteinizing hormone levels, but this did not reach statistical significance. The lower levels of total and calculated free T observed in subjects treated with statins were also confirmed comparing them with age-waist circumference and CV risk score matched controls. Finally, subjects being treated with statins showed lower prolactin levels when compared to the rest of the sample. Our data demonstrated that statin therapy might induce an overt primary hypogonadism and should be considered as a possible confounding factor for the evaluation of testosterone levels in patients with ED.


Polyhydroxyalkanoates as biomaterials.
Polyhydroxyalkanoates (PHAs) are biopolymers synthesized by bacteria under unbalanced growth conditions. These biopolymers are considered as potential biomaterials for future applications because they are biocompatible, biodegradable, and easy to produce and functionalize with strong mechanical strength. Currently, PHAs are being extensively innovated for biomedical applications due to their prerequisite properties. The wide range of biomedical applications includes drug delivery systems, implants, tissue engineering, scaffolds, artificial organ constructs, etc. In this article we review the utility of PHAs in various forms (bulk/nano) for biomedical applications so as to bring about the future vision for PHAs as biomaterials for the advancement of research and technology.


Ribonucleotide reductase inhibitors and future drug design.
Ribonucleotide reductase (RR) is a multisubunit enzyme responsible for the reduction of ribonucleotides to their corresponding deoxyribonucleotides, which are building blocks for DNA replication and repair. The key role of RR in DNA synthesis and cell growth control has made it an important target for anticancer therapy. Increased RR activity has been associated with malignant transformation and tumor cell growth. Efforts for new RR inhibitors have been made in basic and translational research. In recent years, several RR inhibitors, including Triapine, Gemcitabine, and GTI-2040, have entered clinical trial or application. Furthermore, the discovery of p53R2, a p53-inducible form of the small subunit of RR, raises the interest to develop subunit-specific RR inhibitors for cancer treatment. This review compiles recent studies on (1) the structure, function, and regulation of two forms of RR; (2) the role in tumorigenesis of RR and the effect of RR inhibition in cancer treatment; (3) the classification, mechanisms of action, antitumor activity, and clinical trial and application of new RR inhibitors that have been used in clinical cancer chemotherapy or are being evaluated in clinical trials; (4) novel approaches for future RR inhibitor discovery.


Effects of aging on figure-ground perception: Convexity context effects and competition resolution.
We examined age-related differences in figure-ground perception by exploring the effect of age on Convexity Context Effects (CCE; Peterson & Salvagio, 2008). Experiment 1, using Peterson and Salvagio's procedure and black and white stimuli consisting of 2 to 8 alternating concave and convex regions, established that older adults exhibited reduced CCEs compared to younger adults. Experiments 2 and 3 demonstrated that this age difference was found at various stimulus durations and sizes. Experiment 4 compared CCEs obtained with achromatic stimuli, in which the alternating convex and concave regions were each all black or all white, and chromatic stimuli in which the concave regions were homogeneous in color but the convex regions varied in color. We found that the difference between CCEs measured with achromatic and colored stimuli was larger in older than in younger adults. Our results are consistent with the hypothesis that the senescent visual system is less able to resolve the competition among various perceptual interpretations of the figure-ground relations among stimulus regions.


A cardinal role for cathepsin d in co-ordinating the host-mediated apoptosis of macrophages and killing of pneumococci.
The bactericidal function of macrophages against pneumococci is enhanced by their apoptotic demise, which is controlled by the anti-apoptotic protein Mcl-1. Here, we show that lysosomal membrane permeabilization (LMP) and cytosolic translocation of activated cathepsin D occur prior to activation of a mitochondrial pathway of macrophage apoptosis. Pharmacological inhibition or knockout of cathepsin D during pneumococcal infection blocked macrophage apoptosis. As a result of cathepsin D activation, Mcl-1 interacted with its ubiquitin ligase Mule and expression declined. Inhibition of cathepsin D had no effect on early bacterial killing but inhibited the late phase of apoptosis-associated killing of pneumococci in vitro. Mice bearing a cathepsin D(-/-) hematopoietic system demonstrated reduced macrophage apoptosis in vivo, with decreased clearance of pneumococci and enhanced recruitment of neutrophils to control pulmonary infection. These findings establish an unexpected role for a cathepsin D-mediated lysosomal pathway of apoptosis in pulmonary host defense and underscore the importance of apoptosis-associated microbial killing to macrophage function.


Fluoroquinolone-induced tendinopathy: etiology and preventive measures.
Tendinopathy is a serious health problem and its etiology is not fully elucidated. Among intrinsic and extrinsic predisposing factors of tendinopathy, the impact of therapeutic agents, especially fluoroquinolone (FQ) group antibiotics, is recently being recognized. FQs are potent bactericidal agents widely used in various infectious diseases, including community acquired pneumonia and bronchitis, chronic osteomyelitis, traveler's diarrhea, typhoid fever, shigellosis, chronic bacterial prostatitis, uncomplicated cervical and urethral gonorrhea and prophylaxis of anthrax. FQs have an acceptable tolerability range. However, many lines of evidence for developing tendinitis and tendon rupture during FQ use have resulted in the addition of a warning in patient information leaflets. FQ-induced tendinopathy presents a challenge for the clinician because healing response is poor due to low metabolic rate in mature tendon tissue and tendinopathy is more likely to develop in patients who are already at high risk, such as elderly, solid organ transplant recipients and concomitant corticosteroid users. FQs become photo-activated under exposure to ultraviolet light, and this process results in formation and accumulation of intracellular reactive oxygen species (ROS). The subsequent FQ-related oxidative stress disturbs mitochondrial functions, leading to apoptosis. ROS overproduction also has direct cytotoxic effects on extracellular matrix components. Understanding the mechanisms of the FQ-associated tendinopathy may enable designing safer therapeutic strategies, hence optimization of clinical response. In this review, we evaluate multi-factorial etiology of the FQ-induced tendinopathy and discuss proposed preventive measures such as antioxidant use and protection from natural sunlight and artificial ultraviolet exposure.


Cord blood DNA methylome in newborns later diagnosed with autism spectrum disorder reflects early dysregulation of neurodevelopmental and X-linked genes.
Autism spectrum disorder (ASD) is a neurodevelopmental disorder with complex heritability and higher prevalence in males. The neonatal epigenome has the potential to reflect past interactions between genetic and environmental factors during early development and influence future health outcomes. We performed whole-genome bisulfite sequencing of 152 umbilical cord blood samples from the MARBLES and EARLI high-familial risk prospective cohorts to identify an epigenomic signature of ASD at birth. Samples were split into discovery and replication sets and stratified by sex, and their DNA methylation profiles were tested for differentially methylated regions (DMRs) between ASD and typically developing control cord blood samples. DMRs were mapped to genes and assessed for enrichment in gene function, tissue expression, chromosome location, and overlap with prior ASD studies. DMR coordinates were tested for enrichment in chromatin states and transcription factor binding motifs. Results were compared between discovery and replication sets and between males and females. We identified DMRs stratified by sex that discriminated ASD from control cord blood samples in discovery and replication sets. At a region level, 7 DMRs in males and 31 DMRs in females replicated across two independent groups of subjects, while 537 DMR genes in males and 1762 DMR genes in females replicated by gene association. These DMR genes were significantly enriched for brain and embryonic expression, X chromosome location, and identification in prior epigenetic studies of ASD in post-mortem brain. In males and females, autosomal ASD DMRs were significantly enriched for promoter and bivalent chromatin states across most cell types, while sex differences were observed for X-linked ASD DMRs. Lastly, these DMRs identified in cord blood were significantly enriched for binding sites of methyl-sensitive transcription factors relevant to fetal brain development. At birth, prior to the diagnosis of ASD, a distinct DNA methylation signature was detected in cord blood over regulatory regions and genes relevant to early fetal neurodevelopment. Differential cord methylation in ASD supports the developmental and sex-biased etiology of ASD and provides novel insights for early diagnosis and therapy.


Lower urinary tract symptom improvement after radical prostatectomy correlates with degree of prostatic inflammation.
To determine if prostatic inflammation at the time of radical prostatectomy (RP) was associated with the International Prostate Symptom Score (IPSS). We performed a proof of principle analytic case control study of patients who underwent RP between January 2005 and August 2008 for lower urinary tract symptoms (LUTS). We reviewed pathology slides of those who had a change of 4 points or greater, as measured by the IPSS and correlated inflammation with change in IPSS. Multivariate linear regression analyses were performed to determine the association of IPSS with degree of inflammation based on the number of inflammatory cells. Of 249 patients, 136 had complete data and 47 (18.8%) underwent pathologic review. The median change in IPSS for the study cohort was -7.0 points compared to +1.0 point for the control cohort. On univariate analysis, the average improvement in IPSS in patients with severe inflammation was (r = -6.02, 95% confidence interval [CI] -11.0 to -1.1, P = .018) after RP. On multivariate analysis, adjusting for age, body mass index (BMI), year of surgery, history of prostatitis, Gleason score, prostate-specific antigen (PSA), prostate weight, and nerve sparing status, only patients with severe prostatic inflammation had significant improvement in their IPSS (r = -5.93, 95% CI -10.81 to -1.04, P = .004). Prostatic inflammation measured in prostatectomy specimens is associated with worse baseline IPSS than matched cohorts. Specifically, severe inflammation is an independent predictor of IPSS improvement at 1 year after RP.


Integrated rapid-diagnostic-test reader platform on a cellphone.
We demonstrate a cellphone-based rapid-diagnostic-test (RDT) reader platform that can work with various lateral flow immuno-chromatographic assays and similar tests to sense the presence of a target analyte in a sample. This compact and cost-effective digital RDT reader, weighing only ~65 g, mechanically attaches to the existing camera unit of a cellphone, where various types of RDTs can be inserted to be imaged in reflection or transmission modes under light-emitting diode (LED)-based illumination. Captured raw images of these tests are then digitally processed (within less than 0.2 s per image) through a smart application running on the cellphone for validation of the RDT, as well as for automated reading of its diagnostic result. The same smart application then transmits the resulting data, together with the RDT images and other related information (e.g., demographic data), to a central server, which presents the diagnostic results on a world map through geo-tagging. This dynamic spatio-temporal map of various RDT results can then be viewed and shared using internet browsers or through the same cellphone application. We tested this platform using malaria, tuberculosis (TB) and HIV RDTs by installing it on both Android-based smartphones and an iPhone. Providing real-time spatio-temporal statistics for the prevalence of various infectious diseases, this smart RDT reader platform running on cellphones might assist healthcare professionals and policymakers to track emerging epidemics worldwide and help epidemic preparedness.


Coregulation of multiple signaling mechanisms in pp60v-Src-induced closure of Cx43 gap junction channels.
Attenuation in gap junctional coupling has consistently been associated with induction of rapid or synchronous cell division in normal and pathological conditions. In the case of the v-src oncogene, gating of Cx43 gap junction channels has been linked to both direct phosphorylation of tyrosines (Y247 and 265) and phosphorylation of the serine targets of Erk1/2 (S255, 279 and 282) on the cytoplasmic C-terminal domain of Cx43. However, only the latter has been associated with acute, rather than chronic, gating of the channels immediately after v-src expression, a process that is mediated through a "ball-and-chain" mechanism. In this study we show that, while ERK1/2 is necessary for acute closure of gap junction channels, it is not sufficient. Rather, multiple pathways converge to regulate Cx43 coupling in response to expression of v-src, including parallel signaling through PKC and MEK1/2, with additional positive and negative regulatory effects mediated by PI3 kinase, distinguished by the involvement of Akt.


Probing domain mobility in a flavocytochrome.
The crystal structures of various different members of the family of fumarate reductases and succinate dehydrogenases have allowed the identification of a mobile clamp (or capping) domain [e.g., Taylor, P., Pealing, S. L., Reid, G. A., Chapman, S. K., and Walkinshaw, M. D. (1999) Nat. Struct. Biol. 6, 1108-1112], which has been proposed to be involved in regulating accessibility of the active site to substrate. To investigate this, we have constructed the A251C:S430C double mutant form of the soluble flavocytochrome c(3) fumarate reductase from Shewanella frigidimarina, to introduce an interdomain disulfide bond between the FAD-binding and clamp domains of the enzyme, thus restricting relative mobility between the two. Here, we describe the kinetic and crystallographic analysis of this double mutant enzyme. The 1.6 A resolution crystal structure of the A251C:S430C enzyme under oxidizing conditions reveals the formation of a disulfide bond, while Ellman analysis confirms its presence in the enzyme in solution. Kinetic analyses with the enzyme in both the nonbridged (free thiol) and the disulfide-bridged states indicate a slight decrease in the rate of fumarate reduction when the disulfide bridge is present, while solvent-kinetic-isotope studies indicate that in both wild-type and mutant enzymes the reaction is rate limited by proton and/or hydride transfer during catalysis. The limited effects of the inhibition of clamp domain mobility upon the catalytic reaction would indicate that such mobility is not essential for the regulation of substrate access or product release.


Artifact removal in Fourier-domain optical coherence tomography with a piezoelectric fiber stretcher.
We describe an artifact removal setup swept-source optical coherence tomography (OCT) system that enables high-speed full-range imaging. We implement a piezoelectric fiber stretcher to generate a periodic phase shift between successive A-scans, thus introducing a transverse modulation. The depth ambiguity is then resolved by performing a Fourier filtering in the transverse direction before processing the data in the axial direction. The dc artifact is also removed. The key factor is that the piezoelectric fiber stretcher can be used to generate discrete phase shifts with a high repetition rate. The proposed experimental setup is a much improved version of the previously reported B-M mode scanning for spectral-domain OCT in that it does not generate additional artifacts. It is a simple and low-cost solution for artifact removal that can easily be applied.


Challenges and opportunities for characterizing cognitive aging across species.
The gradual decline of cognitive ability with age, even in the absence of overt brain disease, is a growing problem. Although cognitive aging is a common and feared accompaniment of the aging process, its underlying mechanisms are not well understood and there are no highly effective means to prevent it. Additional research on cognitive aging is sorely needed, and methods that enable ready translation between human subjects and animal models stand to provide the most benefit. Here and in the six companion pieces in this special issue, we discuss a variety of challenges and opportunities for studying cognitive aging across species. We identify tests of associative memory, recognition memory, spatial and contextual memory, and working memory and executive function as cognitive domains that are age-sensitive and amenable to testing with parallel means in both humans and animal models. We summarize some of the important challenges in using animal models to test cognition. We describe unique opportunities to study cognitive aging in human subjects, such as those provided by recent large-scale initiatives to characterize cognition in large groups of subjects across the lifespan. Finally, we highlight some of the challenges of studying cognitive aging in human subjects.


CYP1A1 genotype modifies the impact of smoking on effectiveness of HAART among women.
We have recently shown that cigarette smoking is associated with lesser responses to potent antiretroviral therapies. Certain Cytochrome P-450 enzymes activate compounds derived from tobacco smoke into toxic forms that may promote HIV-1 gene expression through promotion of DNA-adduct formation by the oxidation of chemical constituents of cigarette smoke, such as polyaromatic hydrocarbons and dioxins. To explore the association between environmental and genetic factors to viral replication in women who smoke and receive highly active anti-retroviral therapy (HAART), we assessed the impact of polymorphisms in a panel of four Cytochrome P-450 genes (CYP1A1, CYP2A6, CYP2D6, and CYP2E1) and two Glutathione S-transferase genes (GSTM1 and GSST1) in 924 participants of the Women's Interagency HIV Study (WIHS). Our findings showed that GSTM1 and GSST1 deletions were not associated with HAART effectiveness. By contrast, homozygosity for the CYP1A1-m1 polymorphism, was associated with impaired viral response to treatment among smokers (relative hazard (RH) = 0.54; 95% confidence interval = 0.31-0.94) after adjustment for pretreament viral load, CD4 count, age, hepatitis C infection, prior HAART therapy and race, although it had no effect among nonsmokers. We conclude that the association of the CPY1A1-m1 variant with a reduced response to HAART therapy in HIV infected smokers is consistent with this enzyme's role in the metabolic conversion of environmental toxins to DNA adducts, which may directly promote HIV-1 gene expression.


Early treatment failure in intermediate-risk rhabdomyosarcoma: results from IRS-IV and D9803--a report from the Children's Oncology Group.
The goal of this study was to determine the frequency and clinical features of early treatment failure during induction chemotherapy before protocol radiation therapy for children with intermediate-risk rhabdomyosarcoma (RMS). Patients with intermediate-risk RMS enrolled onto the Intergroup Rhabdomyosarcoma Study-IV and the Children's Oncology Group D9803 study were reviewed for an early treatment failure. Early failure was defined as failure caused by progressive disease, death as a result of progressive disease, or death as a result of other causes occurring fewer than 120 days from study entry. Patients with parameningeal site RMS with high-risk features who received radiation therapy at week 1 were excluded from analysis. Overall survival (OS) was estimated using the Kaplan-Meier method. Fisher's exact test was used to compare differences between groups. Cumulative incidence of progression was estimated. Of 916 patients, 20 (2.2%) were found to have an early disease progression and did not receive planned protocol radiotherapy. Three additional early failures resulted from treatment-related death without progression. Median time to failure was 48 days (range, 7 to 106 days). Nineteen (95%) of the 20 patients experienced progression at their primary site. Five-year OS was 32% (95% CI, 12% to 54%) for patients experiencing an early progression. A small proportion of patients with intermediate-risk RMS suffer an early failure as a result of early progression (2.2%) or treatment-related mortality (0.3%). The majority of patients with early progression had a local failure. Earlier radiotherapy could potentially improve outcome by preventing early local progression.


A brain-machine-muscle interface for restoring hindlimb locomotion after complete spinal transection in rats.
A brain-machine interface (BMI) is a neuroprosthetic device that can restore motor function of individuals with paralysis. Although the feasibility of BMI control of upper-limb neuroprostheses has been demonstrated, a BMI for the restoration of lower-limb motor functions has not yet been developed. The objective of this study was to determine if gait-related information can be captured from neural activity recorded from the primary motor cortex of rats, and if this neural information can be used to stimulate paralysed hindlimb muscles after complete spinal cord transection. Neural activity was recorded from the hindlimb area of the primary motor cortex of six female Sprague Dawley rats during treadmill locomotion before and after mid-thoracic transection. Before spinal transection there was a strong association between neural activity and the step cycle. This association decreased after spinal transection. However, the locomotive state (standing vs. walking) could still be successfully decoded from neural recordings made after spinal transection. A novel BMI device was developed that processed this neural information in real-time and used it to control electrical stimulation of paralysed hindlimb muscles. This system was able to elicit hindlimb muscle contractions that mimicked forelimb stepping. We propose this lower-limb BMI as a future neuroprosthesis for human paraplegics.


Genomes from uncultivated prokaryotes: a comparison of metagenome-assembled and single-amplified genomes.
Prokaryotes dominate the biosphere and regulate biogeochemical processes essential to all life. Yet, our knowledge about their biology is for the most part limited to the minority that has been successfully cultured. Molecular techniques now allow for obtaining genome sequences of uncultivated prokaryotic taxa, facilitating in-depth analyses that may ultimately improve our understanding of these key organisms. We compared results from two culture-independent strategies for recovering bacterial genomes: single-amplified genomes and metagenome-assembled genomes. Single-amplified genomes were obtained from samples collected at an offshore station in the Baltic Sea Proper and compared to previously obtained metagenome-assembled genomes from a time series at the same station. Among 16 single-amplified genomes analyzed, seven were found to match metagenome-assembled genomes, affiliated with a diverse set of taxa. Notably, genome pairs between the two approaches were nearly identical (average 99.51% sequence identity; range 98.77-99.84%) across overlapping regions (30-80% of each genome). Within matching pairs, the single-amplified genomes were consistently smaller and less complete, whereas the genetic functional profiles were maintained. For the metagenome-assembled genomes, only on average 3.6% of the bases were estimated to be missing from the genomes due to wrongly binned contigs. The strong agreement between the single-amplified and metagenome-assembled genomes emphasizes that both methods generate accurate genome information from uncultivated bacteria. Importantly, this implies that the research questions and the available resources are allowed to determine the selection of genomics approach for microbiome studies.


Prion protein polymerisation triggered by manganese-generated prion protein seeds.
Prion diseases are neurodegenerative diseases that can be transmitted between individuals. The exact cause of these diseases remains unknown. However, one of the key events associates with the disease is the aggregation of a cellular protein, the prion protein. The mechanism of this is still unclear. However, it is likely that the aggregation is trigged by a seeding mechanism in which an oligomer of the prion protein is able to catalyse polymerisation of further prion protein into larger aggregates. We have developed a model of this process using an oligomeric species generated from recombinant protein by exposure to manganese. On fractionation of the seeding species, we estimated that the smallest size the oligomer would be is an octomer. We analysed the catalytic mechanism of the seeding oligomer and its interaction with substrate. Different domains of the protein are necessary for the seeding ability of the prion protein as opposed to those required for it to form a substrate for the polymerisation reaction. Prion seeds formed from different sheep alleles are able to reproduce the characteristics of scrapie in terms of resistance to disease. However, we were also able to generate prion seed from chicken PrP a species where no prion disease is known. Our findings provide an insight into the aggregation process of the prion protein and its potential relation to disease progress.


Dual anchoring of the GRASP membrane tether promotes trans pairing.
GRASP proteins share an N-terminal GRASP domain and mediate homotypic tethering of Golgi cisternae to form extended Golgi ribbons. The golgin GM130 is thought to bind the C-terminal side of the GRASP domain to recruit GRASP65 onto the Golgi whereas stable membrane association appears to also depend on anchoring of the N terminus by myristoylation. Here, we examine the nature of the GM130/GRASP65 interaction and test whether the dual membrane contacts of the GRASP domain have a role in tethering beyond membrane recruitment. GM130 was found to contain a C-terminal PDZ ligand that binds the putative groove of the second PDZ-like domain in GRASP65. To test tethering activity independent of targeting, we took advantage of a tethering assay carried out on the mitochondrial membrane in which the GRASP membrane attachment points were individually or simultaneously substituted with mitochondrially targeted transmembrane sequences. N-terminally anchored constructs tethered only if the C terminus was also anchored; and likewise, C-terminally anchored constructs tethered only if the N terminus was anchored. One explanation for the role of this dual anchoring is that it orients the GRASP domain to prevent cis interactions within the same membrane thereby favoring trans interactions between adjacent membranes. Indeed, singly anchored GRASP constructs, although nonfunctional in tethering, interacted with one another and also bound and inhibited dually anchored constructs. This work thus elucidates the GM130/GRASP65 interaction and supports a novel orientation-based model of membrane tether regulation in which dual membrane contact orients the tethering interaction interface to favor trans over cis interactions.


Feasibility of radioimmunotherapy of experimental pneumococcal infection.
Streptococcus pneumoniae is an important cause of community-acquired pneumonia, meningitis, and bacteremia. The problem of pneumococcal disease is exacerbated by increasing drug resistance. Furthermore, patients with impaired immunity are at high risk for invasive pneumococcal infections. Thus, there is an urgent need for new approaches to antimicrobial therapy. Antibody therapies take advantage of the specificity and high affinity of the antigen-antibody interaction to deliver antibacterial compounds to a site of infection in the form of naked or conjugated antibodies. We have recently established that radioimmunotherapy (RIT) can be used to treat experimental fungal infections in mice. In the present study, we investigated the feasibility of applying a RIT approach to the treatment of S. pneumoniae infection by evaluating the susceptibility of S. pneumoniae to radiolabeled antibody in vitro and in an animal infection model. For the specific antibody carrier, we used human monoclonal antibody D11, which binds to pneumococcal capsular polysaccharide 8. We have selected the alpha particle emitter (213)Bi as the radionuclide for conjugation to the antibody. Incubation of serotype 8 S. pneumoniae with (213)Bi-D11 resulted in dose-dependent killing of bacteria. RIT of S. pneumoniae infection in C57BL/6 mice showed that 60% more mice survived in the (213)Bi-D11-treated group (80 micro Ci) than in the untreated group (P < 0.01). The treatment did not cause hematological toxicity, as demonstrated by platelet counts. This feasibility study establishes that RIT can be applied to the treatment of bacterial infections.


Green Formation of Novel Pyridinyltriazole-Salicylidene Schiff Bases.
In this work, water was used as solvent for the eco-friendly synthesis of imines under microwave irradiation. In the first step of the study, 5-pyridinyl-3-amino-1,2,4-triazole hydrochlorides were synthesized in the reaction of amino guanidine hydrochloride with different pyridine carboxylic acids under acid catalysis. A green method for 5-pyridinyl-3-amino-1,2,4-triazoles was developed with the assistance of microwave synthesis. In the second step, the eco-friendly synthesis of imines was achieved by reacting 5- pyridinyl-2H-1,2,4-triazol-3-amine hydrochlorides with salicylic aldehyde derivatives to produce 2-(5- pyridinyl-2H-1,2,4-triazol-3-ylimino)methyl)phenol imines. Microwave experiments were done using a monomode Anton Paar Monowave 300 microwave reactor (2.45 GHz). Reaction temperatures were monitored by an IR sensor. Microwave experiments were carried out in sealed microwave process vials G10 with maximum reaction volume of 10 mL. When alternative methods were used, it was impossible to obtain good yields from ethanol. Nevertheless, the use of water was successful for this reaction. After 1-h microwave irritation, a yellow solid was obtained in 82% yield. In this work an eco-friendly protocol for the synthesis of Schiff bases from 5-(pyridin-2-, 3- or 4- yl)-3-amino-1,2,4-triazoles and substituted salicylic aldehydes in water under microwave irradiation was developed. Under the found conditions the high yields for the products were achieved at short reaction time and with an easy isolation procedure.


Causal effect of education on mortality in a quasi-experiment on 1.2 million Swedes.
In 1949-1962, Sweden implemented a 1-y increase in compulsory schooling as a quasi-experiment. Each year, children in a number of municipalities were exposed to the reform and others were kept as controls, allowing us to test the hypothesis that education is causally related to mortality. We studied all children born between 1943 and 1955, in 900 Swedish municipalities, with control for birth-cohort and area differences. Primary outcome measures are all-cause and cause-specific mortality until the end of 2007. The analyses include 1,247,867 individuals, of whom 92,351 died. We found lower all-cause mortality risk in the experimental group after age 40 [hazard ratio (HR) = 0.96, 95% confidence interval (CI) 0.93-0.99] but not before (HR = 1.03, 95% CI 0.98-1.07) or during the whole follow-up (HR = 0.98, 95% CI 0.95-1.01). After age 40, the experimental group had lower mortality from overall cancer, lung cancer, and accidents. In addition, exposed women had lower mortality from ischemic heart disease, and exposed men lower mortality from overall external causes. In analyses stratified for final educational level, we found lower mortality in the experimental group within the strata that settled for compulsory schooling only (HR = 0.94, 95% CI 0.89-0.99) and compulsory schooling plus vocational training (HR = 0.92, 95% CI 0.88-0.97). Thus, the experimental group had lower mortality from causes known to be related to education. Lower mortality in the experimental group was also found among the least educated, a group that clearly benefited from the reform in terms of educational length. However, all estimates are small and there was no evident impact of the reform on all-cause mortality in all ages.


Lower extremity arterial injury patterns and reconstructive outcomes in patients with severe lower extremity trauma: a 26-year review.
Management of severe traumatic lower extremity injuries remains a considerable challenge. Free tissue transfer is now a standard part of reconstruction for Gustilo IIIB and IIIC injuries. There is limited information on arterial injury patterns in this population. We undertook a review of our experience to gain insight on vascular injury patterns and surgical outcomes. A 26-year retrospective analysis was performed of all lower extremity Gustilo IIIB and IIIC injuries requiring microvascular reconstruction at New York University Medical Center. Patient demographics, Gustilo classification, angiographic findings (conventional/computed tomographic angiography/magnetic resonance angiography), recipient vessels, elapsed time from injury, flap choices, and outcomes were examined. Two hundred twenty-two free flaps on 191 patients were performed from September 1982 until March 2008. There were 151 males and 40 females ranging in age from 4 to 83 years (median age 33 years). Patients sustained either Gustilo IIIB (170 patients) or IIIC (21 patients) open fractures. One hundred fifty-four patients had angiograms (78.2% IIIB, 100% IIIC). Sixty-six (42.9%) had normal 3-vessel runoff and 88 (57.1%) were abnormal. Sixty-one patients (31.9%) had anterior tibial injuries, 17 patients (8.9%) had posterior tibial injuries, and 30 (15.7%) had peroneal injuries. Sixty-three complications occurred (11 early thrombosis, 33 requiring secondary procedures, and 10 requiring amputation). Angiography of severe lower extremity injuries requiring free flap reconstruction usually revealed arterial injury and is generally indicated. In our experience, the anterior tibial artery is most commonly injured and the posterior tibial artery is most likely to be spared and used as a recipient.


The Reciprocity between Radiotherapy and Cancer Immunotherapy.
The clinical success of immune checkpoint inhibitors in treating metastatic and refractory cancers has generated significant interest in investigating their role in treating locally advanced diseases, thus requiring them to be combined with standard treatments in the hope of producing synergistic antitumor responses. Radiotherapy, in particular, has long been hypothesized to have actions complementary to those of immune checkpoint blockade, and a growing body of evidence indicates that cancer immunotherapy may also have radiosensitizing effects, which would provide unique benefit for locoregional treatments. Recent studies have demonstrated that when immune cells are activated by immunotherapeutics, they can reprogram the tumor microenvironment in ways that may potentially increase the radiosensitivity of the tumor. In this review, we highlight the evidence that supports reciprocal interactions between cancer immunotherapy and radiotherapy, where in addition to the traditional notion that radiation serves to enhance the activation of antitumor immunity, an alternative scenario also exists in which T-cell activation by cancer immunotherapy may sensitize tumors to radiation treatment through mechanisms that include normalization of the tumor vasculature and tissue hypoxia. We describe the empirical observations from preclinical models that support such effects and discuss their implications for future research and trial design.


Possible Role of Phosphatidylglycerol-Activated Protein Kinase C-βII in Keratinocyte Differentiation.
The epidermis is a continuously regenerating tissue maintained by a balance between proliferation and differentiation, with imbalances resulting in skin disease. We have previously found that in mouse keratinocytes, the lipid-metabolizing enzyme phospholipase D2 (PLD2) is associated with the aquaglyceroporin, aquaporin 3 (AQP3), an efficient transporter of glycerol. Our results also show that the functional interaction of AQP3 and PLD2 results in increased levels of phosphatidylglycerol (PG) in response to an elevated extracellular calcium level, which triggers keratinocyte differentiation. Indeed, we showed that directly applying PG can promote keratinocyte differentiation. We hypothesized that the differentiative effects of this PLD2/AQP3/PG signaling cascade, in which AQP3 mediates the transport of glycerol into keratinocytes followed by its PLD2-catalyzed conversion to PG, are mediated by protein kinase CβII (PKCβII), which contains a PG-binding domain in its carboxy-terminus. Method: To test this hypothesis we used quantitative RT-PCR, western blotting and immunocytochemistry. We first verified the presence of PKCβII mRNA and protein in mouse keratinocytes. Next, we found that autophosphorylated (activated) PKCβII was redistributed upon treatment of keratinocytes with PG. In the unstimulated state phosphoPKCβII was found in the cytosol and perinuclear area; treatment with PG resulted in enhanced phosphoPKCβII localization in the perinuclear area. PG also induced translocation of phosphoPKCβII to the plasma membrane. In addition, we observed that overexpression of PKCβII enhanced calcium- and PG-induced keratinocyte differentiation without affecting calcium-inhibited keratinocyte proliferation. These results suggest that the PG produced by the PLD2/AQP3 signaling module may function by activating PKCβII.


Can application of universal primers alone be a substitute for airborne-particle abrasion to improve adhesion of resin cement to zirconia?
To determine whether universal primers alone can deliver similar levels of adhesion of resin cement to zirconia ceramic when compared to their application in conjunction with airborne-particle abrasion. Sintered zirconia blocks (N = 160) (Lava, 3M ESPE), (5.25 × 5.25 × 3 mm3) were embedded in acrylic resin, polished, and randomly distributed into 16 groups (n = 10 per group), according to the factors "universal primer" (8 levels) and "air-particle abrasion" (2 levels): 1. ctr: control, without application of a universal primer; 2. AP: Alloy Primer; 3. MP: Monobond Plus; 4. MZP: Metal Zirconia Primer; 5. MZ: MZ Primer; 6. Sg: Signum Zirconia Bond; 7. SbU: Singlebond Universal; 8. ZP: Z Prime Plus. The universal primers were also used after air abrasion (A) of zirconia to form the following 8 groups: Ctr-A, AP-A, MP-A, MZP-A, MZ-A, Sg-A, SbU-A, and ZP-A. After ultrasonic cleaning, air abrasion was performed using Al2O3 particles (110 μm, 2.5 bar, 20 s at 10 mm) in a chairside air-abrasion device. After ultrasonic cleaning again, universal primers were applied according to each manufacturer's recommendation. The resin cement (RelyX ARC, 3M ESPE) was built up incrementally and photopolymerized on the zirconia surface using a silicone mold (Ø = 3.5, height = 3 mm). All specimens were stored in distilled water (60 days at 37°C) and then subjected to shear bond strength testing (SBS) in a universal testing machine (1 mm/min). On a separate set of zirconia specimens, contact angle measurements were made using the sessile drop technique with a goniometer after the application of universal primers on control and air-abraded zirconia surfaces. Data (MPa) were analyzed using one-way ANOVA, Tukey's test, and Student's t-test (α = 0.05). When universal primers were used alone, SbU presented significantly higher mean SBS (19.5 ± 5.8) that did the other primers (0 to 9.9 ± 6.6) (p = 0.001). When air abraded, the groups AP-A (14.1 ± 6.1), MP-A (15.9 ± 5.4), ZP-A (16.9 ± 7.3), SG-A (19.1 ± 2.1), SbU-A (12 ± 1.5) showed significant differences (p = 0.03). Adhesive performance of all universal primers was enhanced after air abrasion, with the exception of the SbU and MZ primers. After air abrasion, contact angle measurements were lower for the each primer (without air abrasion: 28.9 to 83.9; with air abrasion: 27.1 to 63.0), except for MZP. Air abrasion with 110 μm Al2O3 followed by universal primer application increased the bond strength of tested resin cement to zirconia, with the exception of SbU and MZ.


Chemistry, Nutrition, and Health-Promoting Properties of Hericium erinaceus (Lion's Mane) Mushroom Fruiting Bodies and Mycelia and Their Bioactive Compounds.
The culinary and medicinal mushroom Hericium erinaceus is widely consumed in Asian countries, but apparently not in the United States, for its nutritional and health benefits. To stimulate broader interest in the reported beneficial properties, this overview surveys and consolidates the widely scattered literature on the chemistry (isolation and structural characterization) of polysaccharides and secondary metabolites such as erinacines, hericerins, hericenones, resorcinols, steroids, mono- and diterpenes, and volatile aroma compounds, nutritional composition, food and industrial uses, and exceptional nutritional and health-promoting aspects of H. erinaceus. The reported health-promoting properties of the mushroom fruit bodies, mycelia, and bioactive pure compounds include antibiotic, anticarcinogenic, antidiabetic, antifatigue, antihypertensive, antihyperlipodemic, antisenescence, cardioprotective, hepatoprotective, nephroprotective, and neuroprotective properties and improvement of anxiety, cognitive function, and depression. The described anti-inflammatory, antioxidative, and immunostimulating properties in cells, animals, and humans seem to be responsible for the multiple health-promoting properties. A wide range of research advances and techniques are described and evaluated. The collated information and suggestion for further research might facilitate and guide further studies to optimize the use of the whole mushrooms and about 70 characterized actual and potential bioactive secondary metabolites to help prevent or treat human chronic, cognitive, and neurological diseases.


Thioalkalimicrobium cyclicum sp. nov. and Thioalkalivibrio jannaschii sp. nov., novel species of haloalkaliphilic, obligately chemolithoautotrophic sulfur-oxidizing bacteria from hypersaline alkaline Mono Lake (California).
Two strains of haloalkaliphilic, obligately autotrophic, sulfur-oxidizing bacteria were isolated from the oxygen-sulfide interface water layer of stratified alkaline and saline Mono Lake, California, USA. Strain ALM 1T was a dominant species in enrichment on moderate-saline, carbonate-buffered medium (0.6 M total Na+, pH 10) with thiosulfate as an energy source and nitrate as a nitrogen source. Cells of ALM 1T are open ring-shaped and are non-motile. It has a high growth rate and activity of thiosulfate and sulfide oxidation and very low sulfur-oxidizing activity. Genetic comparison and phylogenetic analysis suggested that ALM 1T (= DSM 14477T = JCM 11371T) represents a new species of the genus Thioalkalimicrobium in the gamma-Proteobacteria, for which the name Thioalkalimicrobium cyclicum sp. nov. is proposed. Another Mono Lake isolate, strain ALM 2T, dominated in enrichment on a medium containing 2 M total Na+ (pH 10). It is a motile vibrio which tolerates up to 4 M Na+ and produces a membrane-bound yellow pigment. Phylogenetic analysis placed ALM 2T as a member of genus Thioalkalivibrio in the gamma-Proteobacteria, although its DNA hybridization with the representative strains of this genus was only about 30%. On the basis of genetic and phenotypic properties, strain ALM 2T (= DSM 14478T = JCM 11372T) is proposed as Thioalkalivibrio jannaschii sp. nov..


Increased risk of respiratory illness associated with kerosene fuel use among women and children in urban Bangalore, India.
Kerosene is a widely used cooking and lighting fuel in developing countries. The potential respiratory health effects of cooking with kerosene relative to cooking with cleaner fuels such as liquefied petroleum gas (LPG) have not been well characterised. We sampled 600 households from six urban neighbourhoods in Bangalore, India. Each household's primary cook, usually the woman of the house, was interviewed to collect information on current domestic fuel use and whether there was any presence of respiratory symptoms or illness in her or in the children in the household. Our analysis was limited to 547 adult females (ages 18-85) and 845 children (ages 0-17) in households exclusively cooking with either kerosene or LPG. We investigated the associations between kerosene use and the likelihood of having respiratory symptoms or illness using multivariate logistic regression models. Among adult women, cooking with kerosene was associated with cough (OR=1.88; 95% CI 1.19 to 2.99), bronchitis (OR=1.54; 95% CI 1.00 to 2.37), phlegm (OR=1.51; 95% CI 0.98 to 2.33) and chest illness (OR=1.61; 95% CI 1.02 to 2.53), relative to cooking with LPG in the multivariate models. Among children, living in a household cooking with kerosene was associated with bronchitis (OR=1.91; 95% CI 1.17 to 3.13), phlegm (OR=2.020; 95% CI 1.29 to 3.74) and chest illness (OR=1.70; 95% CI 0.99 to 2.90) after adjusting for other covariates. We also found associations between kerosene use and wheezing, difficulty breathing and asthma in adults and cough and wheezing in children, though these associations were not statistically significant. Women and children in households cooking with kerosene were more likely to have respiratory symptoms and illnesses compared with those in households cooking with LPG. Transitioning from kerosene to LPG for cooking may improve respiratory health among adult women and children in this population.