Coordinated transcriptional regulation patterns associated with infertility phenotypes in men.
Microarray gene-expression profiling is a powerful tool for global analysis of the transcriptional consequences of disease phenotypes. Understanding the genetic correlates of particular pathological states is important for more accurate diagnosis and screening of patients, and thus for suggesting appropriate avenues of treatment. As yet, there has been little research describing gene-expression profiling of infertile and subfertile men, and thus the underlying transcriptional events involved in loss of spermatogenesis remain unclear. Here we present the results of an initial screen of 33 patients with differing spermatogenic phenotypes. Oligonucleotide array expression profiling was performed on testis biopsies for 33 patients presenting for testicular sperm extraction. Significantly regulated genes were selected using a mixed model analysis of variance. Principle components analysis and hierarchical clustering were used to interpret the resulting dataset with reference to the patient history, clinical findings and histological composition of the biopsies. Striking patterns of coordinated gene expression were found. The most significant contains multiple germ cell-specific genes and corresponds to the degree of successful spermatogenesis in each patient, whereas a second pattern corresponds to inflammatory activity within the testis. Smaller-scale patterns were also observed, relating to unique features of the individual biopsies.


The prospects for mass population screening in colorectal cancer.
Screening leads to an increased detection of tumours localized to the bowel at the time of surgical treatment which are likely to have an improved prognosis. However, because of the biases associated with screening it would be unwise to advocate a nationwide programme until the results of the controlled trials now in progress are available. At the present time the detection of occult blood in the faeces is the only feasible method of population screening. Haemoccult is a reliable, well tried test but lacks sensitivity. Several new chemical and immunological tests are available but require further evaluation.


Expression of ectopeptidases in scleroderma.
To examine the expression and concentrations of three ectopeptidases likely to be involved in regulating the functional levels of adhesion molecules and the turnover of connective tissue components, in patients with scleroderma (systemic sclerosis) (SSc) and in normal individuals. Monoclonal antibodies against these antigens were used for immunoperoxidase staining of cryostat skin sections and for flow cytometric (fluorescence activated cell sorter) analysis of cultured dermal fibroblasts grown from SSc patients and normal controls. Although neutral endopeptidase-24.11 (NEP) (CD10) was not detected in either SSc or normal skin, aminopeptidase N (APN) (CD13) and dipeptidyl peptidase IV (DPPIV) (CD26) were both readily visualised. However, DPPIV appeared to be present in smaller concentrations in the SSc biopsy specimens. Moreover, while fibroblasts grown in vitro from both SSc and normal skin also had similar concentrations of APN, the expression of DPPIV in the cultured SSc cells was found to be very much less than that present in the normal fibroblasts. It is noteworthy that NEP, which was not detected in the tissue sections, was nevertheless readily detected in fibroblasts in culture. These results show that a number of cell surface proteases are expressed by dermal fibroblasts both in vivo and in vitro, and it is suggested that the marked downregulation of DPPIV in SSc could be at least partly responsible for the increased concentrations of adhesion molecules and matrix proteins associated with the molecular pathology of this disease.


Factors associated with healthy aging in Latin American populations.
Latin American populations may present patterns of sociodemographic, ethnic and cultural diversity that can defy current universal models of healthy aging. The potential combination of risk factors that influence aging across populations in Latin American and Caribbean (LAC) countries is unknown. Compared to other regions where classical factors such as age and sex drive healthy aging, higher disparity-related factors and between-country variability could influence healthy aging in LAC countries. We investigated the combined impact of social determinants of health (SDH), lifestyle factors, cardiometabolic factors, mental health symptoms and demographics (age, sex) on healthy aging (cognition and functional ability) across LAC countries with different levels of socioeconomic development using cross-sectional and longitudinal machine learning models (n = 44,394 participants). Risk factors associated with social and health disparities, including SDH (β > 0.3), mental health (β > 0.6) and cardiometabolic risks (β > 0.22), significantly influenced healthy aging more than age and sex (with null or smaller effects: β < 0.2). These heterogeneous patterns were more pronounced in low-income to middle-income LAC countries compared to high-income LAC countries (cross-sectional comparisons), and in an upper-income to middle-income LAC country, Costa Rica, compared to China, a non-upper-income to middle-income LAC country (longitudinal comparisons). These inequity-associated and region-specific patterns inform national risk assessments of healthy aging in LAC countries and regionally tailored public health interventions.


[Dirichlet's Theorem].
In the introduction to Chapter II of his work L'Intégrale de Fourier et ses Applications à l'Optique, Duffieux reduces Dirichlet's Theorem to a "specific summary." Developed by a convolution where the functions of influence apply [Rev. Opt. 34, 351 (1960)], Dirichlet's Theorem immediately gives data on the form of two functions that the Fourier transform introduces in Fraunhoffer diffraction at infinity. The distribution plane, where one normally cuts off Huygens' pupils, F(x,y), is composed not of points, but of diffraction figures or correlation functions. The function f(u,v) of a spread of frequencies is also a directive function, but it shows no correlation and has discontinuities. F(x,y) is linked with the undulatory theory of light, and f(u,v) represents a corpuscular flux. Two conclusions can be drawn therefrom: (1) the limited pupils must, correctly, be defined by the directive function f(u,v); (2) Fourier's equation establishes a relation between two aspects of the propagation of the lig t crossing a plane, one of which conforms to the undulatory theory of light, the other to his corpuscular theory. We are visibly lacking a corpusculatory theory of light and its coherent images.


Evaluation of a culturally adapted reminiscence therapy intervention: Improving mood, family and community connectedness in Spanish- and Vietnamese-speaking older adults.
Reminiscence therapy (RT) is an evidence-based treatment for alleviating depression and improving life satisfaction among elders, yet less is known about its efficacy in diverse ethnic groups. We report on the evaluation of a cultural adaptation of the RT intervention that combines reminiscence with three innovative elements: including family members in RT; conducting community events for participants to present their artwork and life-story books created during the intervention; and using ethnically matched bilingual community workers. The 12-week intervention with pre- and post-test assessments was completed by 73 Hispanic and 92 Vietnamese elders (mean age = 75 years; 62% female; mostly foreign-born, with limited English proficiency). Paired t-tests indicated statistically significant improvement in depression, loneliness, and life satisfaction. Sixty-two percent of participants noted improved relationships with family/friends through improved communication and shared understanding of the participant's life story. Fifty percent of respondents participated in a community event and the qualitative comments noted improved connection with their community through listening to others' life experiences and sharing their own. Participant perceptions of ethnically matched community workers were positive, but several indicated they could work with someone from a different ethnic background. Similar to general RT findings, the culturally adapted RT intervention demonstrated improvement in depression, loneliness, and life satisfaction among elders from two different ethnic backgrounds. Implications for future projects are discussed based on the findings and experience of conducting this intervention.


The Olfactory Bulb: An Immunosensory Effector Organ during Neurotropic Viral Infections.
In 1935, the olfactory route was hypothesized to be a portal for virus entry into the central nervous system (CNS). This hypothesis was based on experiments in which nasophayngeal infection with poliovirus in monkeys was prevented from spreading to their CNS via transection of olfactory tracts between the olfactory neuroepithelium (ONE) of the nasal cavity and the olfactory bulb (OB). Since then, numerous neurotropic viruses have been observed to enter the CNS via retrograde transport along axons of olfactory sensory neurons whose cell bodies reside in the ONE. Importantly, this route of infection can occur even after subcutaneous inoculation of arboviruses that can cause encephalitis in humans. While the olfactory route is now accepted as an important pathway for viral entry into the CNS, it is unclear whether it provides a way for infection to spread to other brain regions. More recently, studies of antiviral innate and adaptive immune responses within the olfactory bulb suggest it provides early virologic control. Here we will review the data demonstrating that neurotropic viruses gain access to the CNS initially via the olfactory route with emphasis on findings that suggest the OB is a critical immunosensory effector organ that effectively clears virus.


Different expression pattern of human cytomegalovirus-encoded microRNAs in circulation from virus latency to reactivation.
Human cytomegalovirus (HCMV) is a beta-hersvirinae that has a high latent infection rate worldwide and can cause serious consequences in immunocompromised patients when reactivation; however, the mechanism of how HCMV convert from latent to reactivation has rarely been investigated. In the present study, we aimed to perform a comprehensive analysis of the HCMV-encoded microRNA (miRNA) profile in serum of patients upon HCMV reactivation from latency and to further evaluate its clinical significance for the disease monitoring and preventing usefulness. Serum samples from 59 viremia patients and 60 age-gender matched controls were enrolled in this study for screening and validation of different expression of HCMV miRNAs. Serum concentrations of 22 known HCMV miRNAs were determined by a hydrolysis probe-based stem-loop quantitative reverse transcription polymerase chain reaction (RT-qPCR) assay. HCMV DNA was measured by quantitative real-time PCR (qPCR) with the whole blood sample. Serum HCMV IgG and IgM were assessed using enzyme linked immunosorbent assay (ELISA). Another 47 samples from 5 patients at different time points were collected to evaluate the monitoring effectiveness and disease prediction ability of differential expression HCMV-miRNAs during the antiviral treatment. The RT-qPCR analysis revealed that the serum levels of 16 of the 22 examined HCMV miRNAs were elevated in HCMV viremia patients compared with controls, and a profile of 8 HCMV miRNAs including hcmv-miR-US25-2-3p, hcmv-miR-US4-5p, hcmv-miR-US25-2-5p, hcmv-miR-US25-1-3p, hcmv-miR-US25-1, hcmv-miR-UL36, hcmv-miR-UL148D, hcmv-miR-US29-3p were markedly elevated (fold change > 2, P < 0.01). Receiver operating characteristic curve (ROC) analysis were performed on the selected HCMV-miRNAs in all of the patients and controls that enrolled in this study, and which ranged from 0.72 to 0.80 in the autoimmune patients. In addition, hcmv-miR-US25-1-3p levels were significantly correlated with HCMV DNA load (r = 0.349, P = 0.007), and were obviously higher in the reactivation set than the latency set in the autoimmune patients, which could be a predictor for the monitoring of the antiviral treatment. HCMV miRNAs profile showed markedly shift-switch from latency to reactivation in circulation from HCMV infected patients and hcmv-miR-US25-1-3p may be served as a predictor for the switch upon reactivation from latency in patients suffered with autoimmune diseases.


Sequential regulation of developmental events during polar morphogenesis in Caulobacter crescentus: assembly of pili on swarmer cells requires cell separation.
Pili, along with the flagellum and DNA bacteriophage receptors, are structural markers for polar morphogenesis in Caulobacter crescentus. Pili act as primary receptors for a number of small, C. crescentus-specific DNA and RNA bacteriophages, and the timing of pilus-dependent adsorption of bacteriophage phiCb5 in synchronized cell populations has led to the general conclusion that pili are formed coordinately with the flagellum and other polar surface structures in the predivisional cell. The use of rotary platinum shadow casting and electron microscopy as a direct assay for formation of flagella and pili in synchronous cell cultures now shows, however, that when expressed as fractions of the swarmer cell cycle, flagella are assembled on the predivisional cells at approximately 0.8 and that pili are assembled on the new swarmer cells at approximately 0.1 of the next cell cycle. Adsorption of pilus-specific bacteriophage phiCb5 prevented the loss of pili from swarmer cells during development, which suggests that these structures are retracted at the time of stalk formation. Examination of temperature-sensitive cell division mutants showed that the assembly of pili depends on completion of cell separation. These results indicate that the stage-specific events required for polar morphogenesis in C. crescentus occur sequentially, rather than coordinately in the cell cycle, and that the timing of these events reflects the order of underlying cell cycle steps.


Weapons of STAT destruction. Interferon evasion by paramyxovirus V protein.
The signal transducer and activator of transcription (STAT) family of proteins function to activate gene transcription downstream of myriad cytokine and growth factor signals. The prototype STAT proteins, STAT1 and STAT2, are required for innate and adaptive antimicrobial immune responses that result from interferon signal transduction. While many viruses have evolved the ability to avoid these antiviral cytokines, the Paramyxoviruses are distinct in their abilities to interfere directly with STAT proteins. Individual paramyxovirus species differ greatly in their precise mechanism of STAT signaling evasion, but a virus-encoded protein called V plays a central role in this process. The theme of V-dependent interferon evasion and its variations provide significant insights into virus-host interactions and viral immune evasion that can help define targets for antiviral drug design. Exposure of the viral weapons of STAT destruction may also be instructive for application to STAT-directed therapeutics for diseases characterized by STAT hyperactivity.


Efficacy of intacs intrastromal corneal ring segment relative to depth of insertion evaluated with anterior segment optical coherence tomography.
To evaluate the effect of implantation depth of Intacs microthin prescription inserts (Addition Technology Inc, Fremont, California) on visual and topographic outcomes in patients with post- laser-assisted in situ keratomileusis (LASIK) ectasias. Retrospective, observational case series. In this case series, 16 eyes of 12 patients were evaluated. All cases were post-LASIK ectasia that had undergone intrastromal corneal ring segment (ICRS) implantation. The planned insertion depth was 70% of stromal thickness using a manual dissector. At least 12 months postoperatively, all eyes underwent Visante (Carl Zeiss Meditec) AS-OCT to determine insertion depth. Cases were categorized into 3 groups based on the measured implantation depth: 40-59% thickness; 60-79% thickness; and ≥80% thickness. Visual, refractive and topographic outcomes were evaluated relative to implantation depth. The lowest improvement in the study parameters ocurred when the implantation depth was ≥80%. In this group, uncorrected visual acuity (UCVA) and best spectacle corrected VA (BSCVA) improved less than 0.5 lines. Manifest refractive spherical equivalent (MRSE) and mean keratometry (Km) change was less than 0.5 diopters (D). The greatest improvements were observed with implantation depth of 60-79% where UCVA and BSCVA increased by 4.5 and 2.5 lines respectively, and MRSE and Km changed by approximately 2.00 D. Less improvement was found when ICRS were implanted between 40-59% of stromal thickness. Implantation of ICRS greater than 80% of stromal thickness may have no effect on visual and topographic status.


Efficacy of rituximab in thymoma associated minimal change disease: case report.
Thymomas have been associated with a broad spectrum of autoimmune diseases. Minimal change disease (MCD) is the most frequent pathological lesion reported. Pathophysiological mechanisms involved in secondary MCD, and linking MCD to thymoma are not yet fully explained, although the hypothesis of T cell dysfunction has been suggested. The fundamental therapeutic principles are steroids and surgical treatment of thymoma, but failures and relapses often require immunosuppressant combinations. A 62-year-old female was admitted in our unit for a nephrotic syndrome associated with a thymoma. The diagnosis of thymoma associated MCD was confirmed by kidney biopsy. After surgical resection of the thymoma and steroid therapy, no remission was observed. Immunosuppressive therapy was then intensified with introduction of rituximab. Here, we report a steroid-resistant nephrotic syndrome secondary to MCD associated thymoma, which achieved complete remission after rituximab therapy. To the best of our knowledge, this is the first report of the use and efficacy of rituximab therapy in this pathology. Our case report suggests that primary and secondary MCD may share similar pathophysiological mechanisms. It does not allow us to draw any conclusions about the mechanism of action of rituximab, but we believe this report argues for the safety and efficacy of rituximab use in thymoma-associated MCD, and therefore constitutes a rationale for future studies.


Mild cognitive impairment and prospective memory: translating the evidence into neuropsychological practice.
There has been a recent rapid development of research characterizing prospective memory performance in mild cognitive impairment (MCI) in older age. However, this body of literature remains largely separated from routine clinical practice in neuropsychology. Furthermore, there is emerging evidence of effective interventions to improve prospective memory performance. Therefore, our objective in this article was to offer a clinical neuropsychological perspective on the existing research in order to facilitate the translation of the evidence-base into clinical practice. By conducting a critical review of the existing research related to prospective memory and MCI, we highlight how this data can be introduced into clinical practice, either within diagnostic assessment or clinical management. Prospective memory is impaired in older adults with MCI, with a pattern of performance that helps with differential diagnosis from healthy aging. Clinical neuropsychologists are encouraged to add prospective memory assessment to their toolbox for diagnostic evaluation of clients with MCI. Preliminary findings of prospective memory interventions in MCI are promising, but more work is required to determine how different approaches translate to increasing independence in everyday life.


Quantitative Correlation Between Hyaluronic Acid Filler and Hyaluronidase.
The hyaluronic acid-based filler (HA filler) is used worldwide in various applications. In particular, the HA filler is used in the plastics and cosmetic medical field for facial rejuvenation and contouring. In this setting, it is injected into the skin or underlying tissue. Complications of HA filler injection have been relieved using hyaluronidase. However, there is no standard dose to adjust for undesirable HA filler lumpness. In this study, the authors tried to analyze any quantitative correlation between HA filler and hyaluronidase. The back of each rat (total 14 rats) was divided into 4 sites. A volume of 0.5 mL HA filler was injected into the subdermal layer at each site and HA filler nodules were created on the dorsum of each rat. Each nodule was allocated to groups 1, 2, 3, and 4 according to the different concentrations of hyaluronidase. As a result, the injected HA filler volume doubled within 4 days of injection, and then decreased slowly thereafter in group 1 (control group with normal saline only). A 30 unit hyaluronidase treatment compensated for the initial volume increase (approximately 30%) with HA filler (0.5 ml) at the fourth day. Sixty units of hyaluronidase reduced the initial volume (0.5 mL) of overinjected or misplaced HA filler on the fourth day. Approximately 90 units of hyaluronidase can reduce to the volume by 0.25 mL (50%) of the injected HA filler on the fourth day. The authors believe that this quantitative analysis of hyaluronidase concentration is helpful to plan the amount of hyaluronidase for correction of HA filler injection errors.


The HSV-1 mechanisms of cell-to-cell spread and fusion are critically dependent on host PTP1B.
All herpesviruses have mechanisms for passing through cell junctions, which exclude neutralizing antibodies and offer a clear path to neighboring, uninfected cells. In the case of herpes simplex virus type 1 (HSV-1), direct cell-to-cell transmission takes place between epithelial cells and sensory neurons, where latency is established. The spreading mechanism is poorly understood, but mutations in four different HSV-1 genes can dysregulate it, causing neighboring cells to fuse to produce syncytia. Because the host proteins involved are largely unknown (other than the virus entry receptor), we were intrigued by an earlier discovery that cells infected with wild-type HSV-1 will form syncytia when treated with salubrinal. A biotinylated derivative of this drug was used to pull down cellular complexes, which were analyzed by mass spectrometry. One candidate was a protein tyrosine phosphatase (PTP1B), and although it ultimately proved not to be the target of salubrinal, it was found to be critical for the mechanism of cell-to-cell spread. In particular, a highly specific inhibitor of PTP1B (CAS 765317-72-4) blocked salubrinal-induced fusion, and by itself resulted in a dramatic reduction in the ability of HSV-1 to spread in the presence of neutralizing antibodies. The importance of this phosphatase was confirmed in the absence of drugs by using PTP1B-/- cells. Importantly, replication assays showed that virus titers were unaffected when PTP1B was inhibited or absent. Only cell-to-cell spread was altered. We also examined the effects of salubrinal and the PTP1B inhibitor on the four Syn mutants of HSV-1, and strikingly different responses were found. That is, both drugs individually enhanced fusion for some mutants and reduced fusion for others. PTP1B is the first host factor identified to be specifically required for cell-to-cell spread, and it may be a therapeutic target for preventing HSV-1 reactivation disease.


Near-UV laser treatment of extrinsic dental enamel stains.
The selective ablation of extrinsic dental enamel stains using a 400-nm laser is evaluated at several fluences for completely removing stains with minimal damage to the underlying enamel. A frequency-doubled Ti:sapphire laser (400-nm wavelength, 60-nanosecond pulse duration, 10-Hz repetition rate) was used to treat 10 extracted human teeth with extrinsic enamel staining. Each tooth was irradiated perpendicular to the surface in a back-and-forth motion over a 1-mm length using an ∼300-µm-diam 10th-order super-Gaussian beam with fluences ranging from 0.8 to 6.4 J/cm(2) . Laser triangulation determined stain depth and volume removed by measuring 3D surface images before and after irradiation. Scanning electron microscopy evaluated the surface roughness of enamel following stain removal. Fluorescence spectroscopy measured spectra of unbleached and photobleached stains in the spectral range of 600-800 nm. Extrinsic enamel stains are removed with laser fluences between 0.8 and 6.4 J/cm(2) . Stains removed on sound enamel leave behind a smooth enamel surface. Stain removal in areas with signs of earlier cariogenic acid attacks resulted in isolated and randomly located laser-induced, 50-µm-diam enamel pits. These pits contain 0.5-µm diam, smooth craters indicative of heat transfer from the stain to the enamel and subsequent melting and water droplet ejection. Ablation stalling of enamel stains is typically observed at low fluences (<3 J/cm(2) ) and is accompanied by a drastic reduction in porphyrin fluorescence from the Soret band. Laser ablation of extrinsic enamel stains at 400 nm is observed to be most efficient above 3 J/cm(2) with minimal damage to the underlying enamel. Unsound underlying enamel is also observed to be selectively removed after irradiation.


Indocyanine Green-Mediated Photodynamic Therapy Reduces Methicillin-Resistant Staphylococcus aureus Drug Resistance.
Methicillin-resistant Staphylococcus aureus (MRSA) skin-wound infections are associated with considerable morbidity and mortality. Indocyanine green (ICG), a safe and inexpensive dye used in clinical imaging, can be activated by near-infrared in photodynamic therapy (PDT) and photothermal therapy (PTT) to effectively kill MRSA. However, how this treatment affects MRSA drug sensitivity remains unknown. The drug-sensitivity phenotypes, bacterial growth rate, and cell-wall thickness of three MRSA strains were analyzed after ICG-PDT. Drug-resistant gene expressions were determined by polymerase chain reaction (PCR) and quantitative reverse transcription (qRT)-PCR. Related protein expressions were examined with immunoblotting. Drug sensitivity was further evaluated in animal models. MRSA that survived the treatment grew faster, and the cell wall became thinner compared to parental cells. These cells became more sensitive to oxacillin, which was partly related to mecA complex gene deletion. Skin necrosis caused by ICG-PDT-treated MRSA infection was smaller and healed faster than that infected with parental cells. With oxacillin therapy, no bacteria could be isolated from mouse lung tissue infected with ICG-PDT-treated MRSA. ICG-PDT drives MRSA toward an oxacillin-sensitive phenotype. It has the potential to develop into an alternative or adjuvant clinical treatment against MRSA wound infections.


Lung tumor response in strain a mice exposed to tobacco smoke: some dose-effect relationships.
Male strain A/J mice were exposed for 5 mo in a whole-body inhalation chamber to 3 different concentrations of a mixture of cigarette sidestream and mainstream smoke (99, 120, and 176 mg/m(3) of total suspended particulate material, TSP). After an additional 4-mo recovery period in air, lung tumor multiplicities and incidences were determined. The two highest smoke concentrations produced significantly more lung tumors than did the low dose and control groups, although the response to the high was slightly less than to the medium dose. Lung tumor incidences were in all three groups significantly higher than in controls. Lung displacement volume was increased in a dose-dependent manner, but morphometric analysis of the tissues failed to provide evidence for airspace enlargement. Plasma cotinine levels were dose-dependent and similar after 1-day and 5-day exposure. The shape of the dose-response curve and a comparison with previous data suggest that cigarette smoke is only a comparatively weak mouse lung carcinogen.


Mechanism of Ba2+ block of M-like K channels of rod photoreceptors of tiger salamanders.
IKx is a voltage-dependent K+ current in the inner segment of rod photoreceptors that shows many similarities to M-current. The depression of IKx by external Ba2+ was studied with whole-cell voltage clamp. Ba2+ reduced the conductance and voltage sensitivity of IKx tail currents and shifted the voltage range over which they appeared to more positive potentials. These effects showed different sensitivities to Ba2+: conductance was the least sensitive (K0.5 = 7.6 mM), voltage dependence intermediate (K0.5 = 2.4 mM) and voltage sensitivity the most sensitive (K0.5 = 0.2 mM). Ca2+, Co2+, Mn2+, Sr2+, and Zn2+ did not have actions comparable to Ba2+ on the voltage dependence or the voltage sensitivity of IKx tail currents. In high K+ (100 mM), the voltage range of activation of IKx was shifted 20 mV negative, as was the tau-voltage relation. High K+ did not prevent the effect of Ba2+ on conductance, but abolished its ability to affect voltage dependence and voltage sensitivity. Ba2+ also altered the apparent time-course of activation and deactivation of IKx. Low Ba2+ (0.2 mM) slowed both deactivation and activation, with most effect on deactivation; at higher concentrations (1-25 mM), deactivation and activation time courses were equally affected, and at the highest concentrations, 5 and 25 mM Ba2+, the time course became faster than control. Rapid application of 5 mM Ba2+ suggested that the time dependent currents in Ba2+ reflect in part the slow voltage-dependent block and unblock of IKx channels by Ba2+. This blocking action of Ba2+ was steeply voltage-dependent with an apparent electrical distance of 1.07. Ba2+ appears to interact with IKx channels at multiple sites. A model which assumes that Ba2+ has a voltage-independent and a voltage-dependent blocking action on open or closed IKx channels reproduced many aspects of the data; the voltage-dependent component could account for both the Ba(2+)-induced shift in voltage dependence and reduction in voltage sensitivity of IKx tail currents.


A New 3-Day Standardized Eyeblink Conditioning Protocol to Assess Extinction Learning From Infancy to Adulthood.
Associative learning can be observed from the neonatal period onward, providing opportunities to examine changes in basic learning and memory abilities. One method that is suitable to study associative learning is classical eyeblink conditioning (EBC) which is dependent on the cerebellum. Extinction learning can be systematically investigated in this paradigm by varying the context during learning and extinction. Because of methodological difficulties and ethical challenges, no studies have compared extinction learning using EBC across human development. Our goal was to test feasibility of a 3-day delay EBC paradigm that can be used from infancy to adulthood. Acceptance/safety was tested especially for infancy by investigating attrition rates and parental report on infant wellbeing. On a paradigm side, we tested if the paradigm leads to successful acquisition and extinction. An air puff served as unconditional stimulus (US) and a tone as conditional stimulus (CS). On day 1 during acquisition, participants received 36 US-CS pairings in context A. On day 2, participants received 12 acquisition trials in context A to consolidate association learning, followed by 48 extinction trials (tone alone presentations) in context B. Renewal was assessed on day 3 and incorporated 12 CS alone trials presented in both the acquisition context and the extinction context. Eyeblink responses were videotaped and coded offline. The protocol was tested with 12-36-months-old infants (N = 72), adolescents (N = 8), and adults (N = 8). Concerning the acceptance/safety side, attrition ranged from 21 to 58% in infant samples due to the complex preparation of the children for the paradigm. However, attrition is equal to or lower than other infant learning paradigms. Parents of infant samples were very interested in the paradigm and reported low levels of infant stress, exhaustion, and negative feelings during the sessions. Data quality was very high, and no participant had to be excluded because of insufficient data. Concerning the paradigm side, participants showed successful acquisition and extinction as a group. The procedure is ethically sound, feasible, tolerated by many infants, and acceptable among parents. The data show successful acquisition and extinction rates, making the paradigm a valuable tool for investigating developmental changes in extinction learning over the lifespan.


Cortactin Modulates Lung Endothelial Apoptosis Induced by Cigarette Smoke.
Cigarette smoke (CS) is the primary cause of Chronic Obstructive Pulmonary Disease (COPD), and an important pathophysiologic event in COPD is CS-induced apoptosis in lung endothelial cells (EC). Cortactin (CTTN) is a cytoskeletal actin-binding regulatory protein with modulation by Src-mediated tyrosine phosphorylation. Based upon data demonstrating reduced CTTN mRNA levels in the lungs of smokers compared to non-smokers, we hypothesized a functional role for CTTN in CS-induced mitochondrial ROS generation and apoptosis in lung EC. Exposure of cultured human lung EC to CS condensate (CSC) led to the rearrangement of the actin cytoskeleton and increased CTTN tyrosine phosphorylation (within hours). Exposure to CS significantly increased EC mitochondrial ROS generation and EC apoptosis. The functional role of CTTN in these CSC-induced EC responses was explored using cortactin siRNA to reduce its expression, and by using a blocking peptide for the CTTN SH3 domain, which is critical to cytoskeletal interactions. CTTN siRNA or blockade of its SH3 domain resulted in significantly increased EC mitochondrial ROS and apoptosis and augmented CSC-induced effects. Exposure of lung EC to e-cigarette condensate demonstrated similar results, with CTTN siRNA or SH3 domain blocking peptide increasing lung EC apoptosis. These data demonstrate a novel role for CTTN in modulating lung EC apoptosis induced by CS or e-cigarettes potentially providing new insights into COPD pathogenesis.


Cost of hospitalisation and length of stay due to hypoglycaemia in patients with diabetes mellitus: a cross-sectional study.
This study aims to estimate the length of stay and hospitalisation cost of hypoglycaemia, and to identify determinants of variation in the length of stay and hospitalisation cost among individual patients with type 1 or 2 diabetes mellitus. A cross-sectional study was conducted using inpatients records for patients with diabetes mellitus who had been hospitalised due to hypoglycaemic events in two private hospitals in Amman, Jordan between January 2009 and May 2017. All hospitalisation costs were inflated to the equivalent costs in 2017. Hospitalisation cost was estimated from the patient's perspective in Jordanian dinars (JOD). Descriptive analyses and correlation between sociodemographic or clinical characteristics with the cost and length of stay were explored. Predictors of hypoglycaemic hospitalisation cost and length of stay were determined using logistic regression. During the study period a total of 126 patients with diabetes mellitus were hospitalised due to an incident of hypoglycaemia. The mean patient age was 64.2 (SD=19.6) years; half were male. Patients admitted for hypoglycaemia stayed in hospital for a median duration of two days (IQR=2 days). The median cost of hospitalisation for hypoglycaemia was 163.2 JOD (USD 230.1) (IQR=216.3 JOD). We found that the Glasgow coma score was positively associated with length of stay (0.345, p=0.008), and older age was correlated with higher hospitalisation cost (0.207, p=0.02). Patients with a family history of diabetes had higher hospitalisation costs and longer duration of stay (0.306 and 0.275, p<0.05). In addition, being a male patient (0.394, p<0.05) and with an absence of smoking history was associated with longer duration of stay (0.456, p<0.01), but not with higher hospitalisation cost. Costs associated with the incidence of hypoglycaemic events are not low and constitute a large cost component of managing and treating diabetes mellitus. Male patients and patients having a family history of diabetes should receive extra care and education on the prevention of hypoglycaemic events, and a treatment de-intensification approach should be considered if necessary, so we can prevent its associated hospitalisation costs and length of stay.


The effect of thymosin fraction 5 on lipid peroxidation in rabbits fed a high-cholesterol diet.
Plasma and erythrocyte lipid levels and susceptibility of erythrocytes to lipid peroxidation were determined in rabbits fed diet containing 2% (w/w) cholesterol, for 3 months. Hypercholesterolemic rabbits had high plasma and erythrocyte lipid peroxide levels as compared to control rabbits. After high-cholesterol diet, the rabbits in the experimental group were divided into two groups. The first group was fed a normal diet for 21 days and the second group was given normal diet plus thymosin F5 injections every other day for the same period. At the end of this period, plasma and erythrocyte lipid peroxide levels were significantly decreased in the group injected with thymosin F5.


Hybridization and introgression in two ecologically dissimilar Fundulus hybrid zones.
Hybridization and introgression appear more common in rapidly evolving groups, suggesting an important role in the evolutionary process. Detailed studies of how extrinsic or intrinsic forces regulate hybridization and introgression have the potential for broadening our understanding of mechanisms generating diversity. Species in the Fundulus notatus species complex have broad overlapping ranges and occur in replicated hybrid zones along predictable stream gradients. Typical hybrid zone structure has Fundulus olivaceus in headwaters, F. notatus downstream, and hybrid zones near confluences or abrupt shifts in habitat. Rarely, the typical upstream-downstream orientation is reversed raising questions as to how hybrid zones are formed and maintained. We used next-generation sequencing data to study hybridization and introgression in hybrid zones in neighboring drainages that differ in orientation (typical and reversed). We predicted extrinsic forces linked to stream gradients would result in noticeable differences between the two. Contrary to predictions, the data indicate the hybrid zones are remarkably similar. We used individual-based simulations to explore the potential role of intrinsic and extrinsic forces in generating and maintaining typical and reversed hybrid zones. Simulation results were consistent with reversed hybrid zones being formed from stochastic processes combined with strong intrinsic forces and weak extrinsic forces.


Structural and functional roles of a conserved proline residue in the alpha2 helix of Escherichia coli thioredoxin.
Proline 40 in Escherichia coli thioredoxin is located close to the redox active site (Cys32-Cys35) within the alpha2 helix. The conservation of this residue among most of the thioredoxins suggests that it could play an important role in the structure and/or function of this protein. We have substituted Pro40 for Ala by using site-directed mutagenesis and expressed the mutant P40A in E.coli. The effects of the mutation on the biophysical and biological properties of thioredoxin have been analyzed and compared with molecular dynamics simulations. Modeling predicted that the replacement of Pro40 by Ala induced a displacement of the active site which exposes Trp31 to the solvent and opens a cleft located between helices alpha2 and alpha3. The solvation free energy (SFE) calculation also indicated that P40A became more hydrophobic as W31 became more accessible. These predictions were totally in agreement with the experimental results. The mutant P40A exhibited chromatographic behavior and fluorescence properties very different from those of the wild-type (WT) protein, in relationship with the displacement of W31. The determination of the free energy of unfolding of P40A showed that the mutant was globally destabilized by 2.9 kcal/mol. However, the effect of the mutation on the transition curve was highly unusual as the midpoint of the unfolding transition increased, indicating that some local structures were actually stabilized by the mutation. Despite these structural modifications, neither the ability of the protein to reduce a chloroplastic enzyme nor its reactivity with the bacterial reductase decreased. The only functional difference was the higher stability of P40A in light activation of NADP-malate dehydrogenase under air, which suggests that the mutant was less rapidly re-oxidized than WT. Therefore, it can be concluded that Pro40 is not essential for maintaining the redox function of thioredoxin but rather is required for the stability of the protein.


Human serum antibody response against Streptococcus mutans antigens.
Antigens from Streptococcus mutans were examined to identify specific polypeptides that may have stimulated antibody responses and possibly play some role in caries immunity. A group of 10 adult human subjects was screened for serum antibodies reactive with antigens from S. mutans. Extracellular and cellular protein preparations from S. mutans LM7 (Bratthall serotype e) and V403 (biotype c) were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by Western electrophoretic transfer and immunoblotting analysis. Antibodies reactive with polypeptides ranging from 34 to 400 kilodaltons in apparent molecular mass were detected by these means. Radioimmunoassay competition experiments revealed that the cellular and extracellular antigens did not compete with each other for serum antibodies. Preabsorption of sera with extracellular proteins from other oral streptococcal species prior to immunoblotting indicated that the antigens unique to S. mutans have molecular masses greater than 100 kilodaltons, and each individual produced antibodies against different antigens of high molecular mass. Examination of sera from young children also indicated heterogeneous responses against S. mutans LM7 antigens.


Thermo-Electro-Responsive Redox-Copolymers for Amplified Solvation, Morphological Control, and Tunable Ion Interactions.
Electro-responsive metallopolymers can possess highly specific and tunable ion interactions, and have been explored extensively as electrode materials for ion-selective separations. However, there remains a limited understanding of the role of solvation and polymer-solvent interactions in ion binding and selectivity. The elucidation of ion-solvent-polymer interactions, in combination with the rational design of tailored copolymers, can lead to new pathways for modulating ion selectivity and morphology. Here, we present thermo-electrochemical-responsive copolymer electrodes of N-isopropylacrylamide (NIPAM) and ferrocenylpropyl methacrylamide (FPMAm) with tunable polymer-solvent interactions through copolymer ratio, temperature, and electrochemical potential. As compared to the homopolymer PFPMAm, the P(NIPAM0.9-co-FPMAm0.1) copolymer ingressed 2 orders of magnitude more water molecules per doping ion when electrochemically oxidized, as measured by electrochemical quartz crystal microbalance. P(NIPAM0.9-co-FPMAm0.1) exhibited a unique thermo-electrochemically reversible response and swelled up to 83% after electrochemical oxidation, then deswelled below its original size upon raising the temperature from 20 to 40 °C, as measured through spectroscopic ellipsometry. Reduced P(NIPAM0.9-co-FPMAm0.1) had an inhomogeneous depth profile, with layers of low solvation. In contrast, oxidized P(NIPAM0.9-co-FPMAm0.1) displayed a more uniform and highly solvated depth profile, as measured through neutron reflectometry. P(NIPAM0.9-co-FPMAm0.1) and PFPMAm showed almost a fivefold difference in selectivity for target ions, evidence that polymer hydrophilicity plays a key role in determining ion partitioning between solvent and the polymer interface. Our work points to new macromolecular engineering strategies for tuning ion selectivity in stimuli-responsive materials.


Enhanced production of LPS-induced cytokines during differentiation of human monocytes to macrophages. Role of LPS receptors.
In vitro cultivated human monocytes isolated from normal peripheral blood show a time-dependent differentiation into macrophages characterized by an increased expression of transferrin receptors, CD11/CD18, and CD14 Ag. We measured the secretion of TNF-alpha and IL-6 in freshly isolated monocytes and in differentiated macrophages after LPS treatment. Differentiated macrophages produced significantly higher amounts of TNF-alpha and IL-6 than freshly isolated monocytes. This increased secretion was not a result of an enhanced accumulation of TNF-alpha and IL-6 mRNA, as comparative levels of these transcripts were found in both cell types after LPS treatment. Furthermore, LPS did not induce an antiviral state to VSV3 in monocytes, but it reduced by 3 to 5 log10 the virus yield in differentiated macrophages. The addition of antibodies to IFN-beta completely inhibited the LPS-induced antiviral state to VSV, but antibodies to IFN-alpha, TNF-alpha, or IL-6 were ineffective. A marked accumulation of IFN-beta mRNA was found in both cell types after LPS treatment. Binding experiments with FITC-LPS revealed a slightly higher overall binding affinity for LPS in freshly explanted monocytes as compared with differentiated macrophages, even though the maximal binding was higher in macrophages. In both cell types, the LPS binding was partially inhibited by antibodies to CD14. These results demonstrate that: 1) in vitro differentiation of human monocytes to macrophages leads to an enhanced LPS response in terms of (a) progressive increase of IL-6/TNF-alpha production and (b) acquisition of an IFN-beta mediated antiviral state; 2) this enhanced response to LPS, largely CD14-independent, is not linked to any increased accumulation of cytokine mRNA, but is probably a result of an increased synthesis and/or secretion of these cytokines.


A structural perspective on the where, how, why, and what of nucleosome positioning.
The DNA in eukaryotic chromatin is packed by histones into arrays of repeating units called nucleosomes. Each nucleosome contains a nucleosome core, where the DNA is wrapped around a histone octamer, and a stretch of relatively unconstrained DNA called the linker DNA. Since nucleosome cores occlude the DNA from many DNA-binding factors, their positions provide important clues for understanding chromatin packing and gene regulation. Here we review the recent advances in the genome-wide mapping of nucleosome positions, the molecular and structural determinants of nucleosome positioning, and the importance of nucleosome positioning in chromatin higher order folding and transcriptional regulation.


[Contralateral reflux after unilateral antireflux surgery].
108 unilateral antireflux surgical procedures are analysed over a period of seven years. All the cases were operated upon for essential or primary renal reflux, using ureteral advancement techniques crossing the bladder midline. There were 10 patients who developed contralateral renal reflux (CRR) in the postoperative evolution (9,2%). The CRR disappeared in all cases, but three of them needed surgical treatment because of urinary tract deterioration (Reflux nephropathy, bladder lithiasis, and shift of reflux grade from I to III). Two different surgical antireflux techniques were utilised. Using the Glenn-Anderson's technique the incidence of CRR was 9/68 (13,2%) whereas this incidence was 1/40 (2,5%) using the Cohen's technique, which does not disrupt the trigonal area. The authors do not believe that systematic bilateral antireflux surgery for unilateral reflux is necessary. When CRR occurs an initially conservative management is recommended.


Role of Lamin A/C Gene Mutations in the Signaling Defects Leading to Cardiomyopathies.
Nuclear lamin A/C are crucial components of the intricate protein mesh that underlies the inner nuclear membrane and confers mainly nuclear and cytosolic rigidity. However, throughout the years a number of other key physiological processes have been associated with lamins such as modulation of both genes expression and the activity of signaling mediators. To further solidify its importance in cell physiology, mutations in the lamin A/C gene (LMNA) have been associated to diverse pathological phenotypes with skeletal muscles and the heart being the most affected systems. When affected, the heart develops a wide array of phenotypes spanning from dilated cardiomyopathy with conduction defects to arrhythmogenic right ventricular cardiomyopathy. The surprising large number of cardiac phenotypes reflects the equally large number of specific mutations identified in the LMNA gene. In this review, we underlie how mutations in LMNA can impact the activity and the spatial/temporal organization of signaling mediators and transcription factors. We analyzed the ever-increasing amount of findings collected in LmnaH222P/H222P mice whose cardiomyopathy resemble the most important features of the disease in humans and a number of key evidences from other experimental models. With this mini review, we attempt to combine the newest insights regarding both the pathogenic effects of LMNA mutations in terms of signaling abnormalities and cardiac laminopathies.


Persistent fetal pulmonary hypoperfusion after acute hypoxia.
To determine the effects of duration of hypoxia on fetal pulmonary blood flow and vasoreactivity, we studied the response of the fetal pulmonary vascular bed before, during, and after prolonged (2-h) and more brief (30-min) exposures to acute hypoxia in 19 chronically instrumented unanesthetized fetal lambs. Left pulmonary arterial blood flow was measured by an electromagnetic flow transducer. Fetal PO2 was lowered by delivering 10-12% O2 to the ewe. During 2-h periods of hypoxia left pulmonary arterial blood flow decreased, and main pulmonary arterial and pulmonary vascular resistance increased. The increase in pulmonary vascular resistance was sustained throughout the 2-h period of hypoxia. After the return of the ewe to room air breathing, pulmonary vascular resistance remained elevated for at least 1 h despite the rapid correction of hypoxemia and in the absence of acidemia. In contrast, after 30 min of hypoxia, left pulmonary arterial blood flow, pulmonary arterial pressure, and pulmonary vascular resistance returned to base-line values rapidly with the termination of hypoxia. The persistent pulmonary hypoperfusion after 2 h of hypoxia was attenuated by alpha-adrenergic blockade and was characterized by a blunted vasodilatory response to increases in fetal PO2. When fetal PO2 was elevated during the posthypoxia period in the presence of alpha-blockade, pulmonary blood flow still remained unresponsive to increases in fetal PO2. We conclude that 2-h periods of acute hypoxia can decrease fetal pulmonary vasoreactivity, and we speculate that related mechanisms may contribute to the failure of the normal adaptation of the pulmonary circulation at birth.


In Vivo Characterization of 4 68Ga-Labeled Multimeric RGD Peptides to Image αvβ3 Integrin Expression in 2 Human Tumor Xenograft Mouse Models.
αvβ3 integrins play an important role in angiogenesis and cell migration in cancer and are highly expressed on the activated endothelial cells of newly formed blood vessels. Here, we compare the targeting characteristics of 4 68Ga-labeled multimeric cyclic arginine-glycine-aspartate (RGD)-based tracers in an αvβ3 integrin-expressing tumor model and a tumor model in which αvβ3 integrin is expressed solely on the neovasculature. Methods: Female BALB/c nude mice were subcutaneously injected with SK-RC-52 (αvβ3 integrin-positive) or FaDu (αvβ3 integrin-negative) tumor cells. 68Ga-labeled DOTA-(RGD)2, TRAP-(RGD)3, FSC-(RGD)3, or THP-(RGD)3 was intravenously administered to the mice (0.5 nmol per mouse, 10-20 MBq), followed by small-animal PET/CT imaging and ex vivo biodistribution studies 1 h after injection. Nonspecific uptake of the tracers in both models was determined by coinjecting an excess of unlabeled DOTA-(RGD)2 (50 nmol) along with the radiolabeled tracers. Results: Imaging and biodistribution data showed specific uptake in the tumors for each tracer in both models. Tumor uptake of 68Ga-FSC-(RGD)3 was significantly higher than that of 68Ga-DOTA-(RGD)2, 68Ga-TRAP-(RGD)3, or 68Ga-THP-(RGD)3 in the SK-RC-52 model but not in the FaDu model, in which 68Ga-FSC-(RGD)3 showed significantly higher tumor uptake than 68Ga-TRAP-(RGD)3 Most importantly, differences were also observed in normal tissues and in tumor-to-blood ratios. Conclusion: All tracers showed sufficient targeting of αvβ3 integrin expression to allow for tumor detection. Although the highest tumor uptake was found for 68Ga-FSC-(RGD)3 and 68Ga-THP-(RGD)3 in the SK-RC-52 and FaDu models, respectively, selection of the optimal tracer for specific diagnostic applications also depends on tumor-to-blood ratio and uptake in normal tissues; these factors should therefore also be considered.


Comparison of Suicidal People Who Use the Internet for Suicide-Related Reasons and Those Who Do Not.
Studies have shown that suicidal individuals who use the Internet for suicide-related purposes are more suicidal than nonsuicide-related users, but evidence from European countries is lacking. We assessed differences between suicidal individuals who go online for suicide-related purposes and other suicidal individuals in Austria. Participants were 53 individuals aged between 18 and 24 years who felt suicidal in the past year and completed an anonymous online survey. Individuals who went online for suicide-related purposes were more suicidal, and more depressed, compared with individuals who did not use the Internet for suicide-related purposes. There were no large differences with regard to social anxiety and perceived barriers for help-seeking between the two groups. This study finds that suicidal online users are more depressed and suicidal than nonusers in a sample from Central Europe. Online interventions to combat depression and suicidality need to be strengthened to reach out to this important target population.


Double-pass versus aberrometric modulation transfer function in green light.
Intraocular scattering can become an important source of optical degradation in the aging eye. To evaluate its relative contribution to the ocular modulation transfer function (MTF), a compact, dual experimental system comprising a laser ray tracing (LRT) wavefront sensor and a double-pass setup is used. An aberrometric MTF is estimated from aberration measurements, whereas a second MTF is derived from the double-pass point-spread function. While the former only accounts for the effect of aberrations (up to seventh order), the double-pass MTF includes the combined effect of both scattering and aberrations. A 532-nm laser light source is used to minimize choroidal scattering. Measurements are done on 19 normal, healthy eyes from three groups of subjects of different ages. The two MTFs are obtained for a 6-mm pupil diameter and partial refractive compensation. Intraocular scattering is modeled as a random wavefront aberration characterized by its variance and correlation length. These parameters are fitted from the differences between both MTFs. Our results show that double-pass and LRT techniques provide similar MTFs for most normal eyes, although small amounts of scattering, or high-order aberrations, could be measured in some eyes. A gradual increase in intraocular scattering with age is also observed.


Hepatitis C virus infection, cryoglobulinemia, and peripheral neuropathy: a case report.
Hepatitis C virus (HCV) is essentially hepatotropic but its manifestations can extend beyond the liver. It can be associated with autoimmune diseases, such as mixed cryoglobulinemia, membranoproliferative glomerulonephritis, autoimmune thyroiditis, and lymphoproliferative disorders. The mechanisms that trigger these manifestations are not completely understood. We describe a 48-year-old man with chronic HCV infection (circulating HCV RNA and moderate hepatitis as indicated by liver biopsy), cryoglobulinemia, and sensory and motor peripheral neuropathy. The diagnosis of multineuropathy was confirmed by clinical examination and electromyographic tests. A nerve biopsy revealed an inflammatory infiltrate in the perineurial space and signs of demyelination and axonal degeneration. The patient had no improvement of neurological symptoms with the use of analgesics and neuro-modulators. He was then treated with interferon-alpha (3 million units subcutaneously, 3 times per week) and ribavirin (500 mg orally, twice a day) for 48 weeks. Six months after the end of therapy, the patient had sustained viral response (negative HCV RNA) and remission of neurological symptoms, but cryoglobulins remained positive. A review of the literature on the pathogenesis and treatment of neurological manifestations associated with HCV infection is presented. This report underscores the need for a thorough evaluation of HCV-infected patients because of the possibility of extrahepatic manifestations. Antiviral treatment with interferon and ribavirin can be effective and should be considered in patients with neurological complications associated with HCV infection.


Experiences of disability consumer-directed care users in Australia: results from a longitudinal qualitative study.
The rapidly growing body of literature suggests that Consumer-directed Care (CDC) has the potential to empower consumers and improve the flexibility and quality of care. However, reports highlighting quality and risk concerns associated with CDC focusing on a longer time frame have been few. This paper presents the findings from a qualitative longitudinal evaluation of an Australian CDC programme. Focusing on the period between 2003 and 2008, it reports on the experiences of 12 families caring for a dependent family member. It is based on two external evaluations completed 6 and 36 months after enrollment, and one internal evaluation completed 48 months after enrollment. The findings were triangulated with internal memos, reports and minutes of meetings, as well as with the theoretical literature. The study demonstrates that CDC harbours considerable benefits for people with disabilities and their carers. However, the study also suggests that, over time, carers may experience an increased sense of isolation and lack of support as a result of their involvement in the CDC programme. The paper argues that the development of safeguards addressing these weaknesses is crucial for the sustainability of CDC programmes in contexts where risk cannot be simply transferred onto consumers.


Eight-year randomized clinical evaluation of Class II nanohybrid resin composite restorations bonded with a one-step self-etch or a two-step etch-and-rinse adhesive.
The aim of this study is to observe the durability of Class II nanohybrid resin composite restorations, placed with two different adhesive systems, in an 8-year follow-up. Seventy-eight participants received at random at least two Class II restorations of the ormocer-based nanohybrid resin composite (Ceram X) bonded with either a one-step self-etch adhesive (Xeno III) or a control two-step etch-and-rinse adhesive (Excite). The 165 restorations were evaluated using slightly modified United States Public Health Services (USPHS) criteria at baseline and then yearly during 8 years. One hundred and fifty-eight restorations were evaluated after 8 years. Three participants with five restorations (three Xeno III, two Excite) were registered as dropouts. Twenty-one failed restorations (13.3%) were observed during the follow-up. Twelve in the one-step self-etch adhesive group (13.5%) and nine in the two-step etch-and-rinse group (13.0%). This resulted in nonsignificant different annual failure rates of 1.69 and 1.63%, respectively. Fracture of restoration was the main reason for failure. Good clinical performance was shown during the 8-year evaluation and no significant difference in overall clinical performance between the two adhesives. Fracture was the main reason for failure. The one-step self-etch adhesive showed a good long-term clinical effectiveness in combination with the nanohybrid resin composite in Class II restorations.


Mammalian thioredoxin is a direct inhibitor of apoptosis signal-regulating kinase (ASK) 1.
Apoptosis signal-regulating kinase (ASK) 1 was recently identified as a mitogen-activated protein (MAP) kinase kinase kinase which activates the c-Jun N-terminal kinase (JNK) and p38 MAP kinase pathways and is required for tumor necrosis factor (TNF)-alpha-induced apoptosis; however, the mechanism regulating ASK1 activity is unknown. Through genetic screening for ASK1-binding proteins, thioredoxin (Trx), a reduction/oxidation (redox)-regulatory protein thought to have anti-apoptotic effects, was identified as an interacting partner of ASK1. Trx associated with the N-terminal portion of ASK1 in vitro and in vivo. Expression of Trx inhibited ASK1 kinase activity and the subsequent ASK1-dependent apoptosis. Treatment of cells with N-acetyl-L-cysteine also inhibited serum withdrawal-, TNF-alpha- and hydrogen peroxide-induced activation of ASK1 as well as apoptosis. The interaction between Trx and ASK1 was found to be highly dependent on the redox status of Trx. Moreover, inhibition of Trx resulted in activation of endogenous ASK1 activity, suggesting that Trx is a physiological inhibitor of ASK1. The evidence that Trx is a negative regulator of ASK1 suggests possible mechanisms for redox regulation of the apoptosis signal transduction pathway as well as the effects of antioxidants against cytokine- and stress-induced apoptosis.


Thermodynamic and kinetic characterization of ligand binding to the purine riboswitch aptamer domain.
Riboswitches are cis-acting genetic regulatory elements found commonly in bacterial mRNAs that consist of a metabolite-responsive aptamer domain coupled to a regulatory switch. Purine riboswitches respond to intracellular concentrations of either adenine or guanine/hypoxanthine to control gene expression. The aptamer domain of the purine riboswitch contains a pyrimidine residue (Y74) that forms a Watson-Crick base-pairing interaction with the bound purine nucleobase ligand that discriminates between adenine and guanine. We sought to understand the structural basis of this specificity and the mechanism of ligand recognition by the purine riboswitch. Here, we present the 2,6-diaminopurine-bound structure of a C74U mutant of the xpt-pbuX guanine riboswitch, along with a detailed thermodynamic and kinetic analysis of nucleobase recognition by both the native and mutant riboswitches. These studies demonstrate clearly that the pyrimidine at position 74 is the sole determinant of purine riboswitch specificity. In addition, the mutant riboswitch binds adenine and adenine derivatives well compared with the guanine-responsive riboswitch. Under our experimental conditions, 2,6-diaminopurine binds the RNA with DeltaH=-40.3 kcal mol(-1), DeltaS=-97.6 cal mol(-1)K(-1), and DeltaG=-10.73 kcal mol(-1). A kinetic determination of the slow rate (0.15 x 10(5)M(-1)s(-1) and 2.1 x 10(5)mM(-1)s(-1) for 2-aminopurine binding the adenine-responsive mutant riboswitch and 7-deazaguanine-binding guanine riboswitch, respectively) of association under varying experimental conditions allowed us to propose a mechanism for ligand recognition by the purine riboswitch. A conformationally dynamic unliganded state for the binding pocket is stabilized first by the Watson-Crick base pairing between the ligand and Y74, and by the subsequent ordering of the J2/3 loop, enclosing the ligand within the three-way junction.


A decrease in the plasma DHEA to cortisol ratio during smoking abstinence may predict relapse: a preliminary study.
Increases in depressive symptoms during smoking cessation have been associated with risk for relapse. Several studies have linked plasma levels of cortisol and dehydroepiandrosterone (DHEA) or DHEA-sulfate (DHEAS) to depressive symptoms. To determine whether changes in plasma cortisol, DHEA, or DHEAS levels and emergence of depressive symptoms during smoking cessation are associated with smoking relapse. Subjects were healthy non-medicated men and women, aged 39+/-12 years, who smoked, on average, 22 cigarettes per day. Depressive symptoms, smoking withdrawal symptoms, and plasma steroid levels were measured before and after 8 days of verified smoking abstinence. Relapse status at day 15 was then determined. In the full sample (n=63), there was a trend for changes in depressive symptoms to be associated with relapse. In the subset of 25 subjects with plasma neuroactive steroid data, there was a significant interaction between the change in the plasma DHEA/cortisol ratio from day 0 to day 8 and relapse status at day 15. This ratio was similar before abstinence, but lower at day 8 in relapsed, compared to abstinent, subjects. Changes in the DHEA/cortisol ratio tended to predict changes in depressive symptoms in the women only. A decrease in the plasma DHEA/cortisol ratio during 8 days of smoking abstinence was associated with relapse over the following week. Further research is needed to fully characterize sex-specific relationships between abstinence-induced changes in neuroactive steroid levels, depressive or withdrawal symptoms, and relapse. Such research may lead to new interventions for refractory smoking dependence.


When the mask falls: the role of facial motor resonance in memory for emotional language.
The recognition and interpretation of emotional information (e.g., about happiness) has been shown to elicit, amongst other bodily reactions, spontaneous facial expressions occurring in accordance to the relevant emotion (e.g. a smile). Theories of embodied cognition act on the assumption that such embodied simulations are not only an accessorial, but a crucial factor in the processing of emotional information. While several studies have confirmed the importance of facial motor resonance during the initial recognition of emotional information, its role at later stages of processing, such as during memory for emotional content, remains unexplored. The present study bridges this gap by exploring the impact of facial motor resonance on the retrieval of emotional stimuli. In a novel approach, the specific effects of embodied simulations were investigated at different stages of emotional memory processing (during encoding and/or retrieval). Eighty participants underwent a memory task involving emotional and neutral words consisting of an encoding and retrieval phase. Depending on the experimental condition, facial muscles were blocked by a hardening facial mask either during encoding, during retrieval, during both encoding and retrieval, or were left free to resonate (control). The results demonstrate that not only initial recognition but also memory of emotional items benefits from embodied simulations occurring during their encoding and retrieval.


Genetic relationships of American alligator populations distributed across different ecological and geographic scales.
Although much work has been conducted on coastal populations of the American alligator (Alligator mississippiensis), less is known about the population dynamics and genetic structure of populations of alligators confined to inland habitats. DNA microsatellite loci, derived from the American alligator, were used to investigate patterns of genetic variation within and between populations of alligators distributed at coastal and inland localities in Texas. These data were used to evaluate the genetic discreteness of different alligator stocks relative to their basic ecology at these sites. Observed mean heterozygosities across seven loci for both coastal and inland populations ranged from 0.50-0.61, with both inland and coastal populations revealing similar patterns of variation. Measures of F(st) revealed significant population differentiation among all populations; however, analyses of molecular variance (AMOVAs) failed to demonstrate any apparent geographic pattern relative to the population differentiation indicated by F(st) values. Each population contained unique alleles for at least one locus. Additionally, assignment tests based on the distribution of genotypes placed 76% of individuals to their source population. These genetic data suggest considerable subdivision among alligator populations, possibly influenced by demographic and life history differences as well as barriers to dispersal. These results have clear implications for management. Rather than managing alligators in Texas as a single panmictic population, translocation programs and harvest quotas should consider the ecological and genetic distinctiveness of local alligator populations.


Modulation of proliferation and differentiation of C2C12 skeletal muscle cells by fatty acids.
This study was performed to elucidate whether mitogen-activated protein kinases (MAPKs) are involved in the modulation of the proliferation and differentiation of skeletal muscle cells by fatty acids. C2C12 myoblasts were cultured in differentiation medium containing 2% horse serum for 3 days, and treated with each fatty acid. Phosphorylation levels of MAPKs were examined by immunoblot analysis. The mono-unsaturated fatty acids (MUFAs), oleic acid (OA) and n-6 polyunsaturated fatty acids (n-6 PUFAs), linoleic acid (LA), gamma-linoleic acid (GLA), and arachidonic acid (AA) increased the proliferation of C2C12 cells. On the other hand, n-3 polyunsaturated fatty acids (n-3 PUFAs) and saturated fatty acids (SFs) did not affect the proliferation of C2C12 cells. In addition, the treatment of cis-9, trans-11 conjugated linoleic acid (c9,t11 CLA) showed an increased cell proliferation. However, trans-10, cis-12 conjugated linoleic acid (t10,c12 CLA) significantly inhibited cell proliferation. Treatment of C2C12 cells with LA, OA, and c9,t11 CLA increased phosphorylation levels of ERK1/2 and JNK during proliferation. During cell differentiation, OA, LA, and c9,t11 CLA stimulated differentiation of C2C12 cells, whereas t10,c12 CLA inhibited differentiation. We also found that OA, LA, and c9, t11 CLA increased phosphorylation level of ERK1/2, but not JNK during differentiation. These results suggest that fatty acids are able to modulate the proliferation and differentiation of skeletal muscle and MAPKs may be involved in the modulation of the proliferation and differentiation of skeletal muscle cells by fatty acids.


Success of mineral trioxide aggregate in pulpotomized primary molars.
The aim of the present study was to compare, clinically and radiographically, the mineral trioxide aggregate (MTA) to formocresol (FC) when used as medicaments in pulpotomized vital human primary molars. The sample consisted of 120 primary molars, all teeth were treated with the same conventional pulpotomy technique. Sixty molars received FC and 60 received MTA throughout a random selection technique. At the end of 24-month evaluation period, 74 molars (36 FC, 38 MTA) were available for clinical and radiographic evaluation. None of the MTA treated teeth showed any clinical or radiographic pathology, while the FC group showed a success rate of 86.8% radiographically and 98.6% clinically. The difference between the two groups in the radiographic outcomes was statistically significant. It was concluded that MTA treated molars demonstrated significantly greater success. MTA seems to be a suitable replacement for formocresol in pulpotomized primary teeth.


Millisecond Timescale Motions Connect Amino Acid Interaction Networks in Alpha Tryptophan Synthase.
Tryptophan synthase is a model system for understanding allosteric regulation within enzyme complexes. Amino acid interaction networks were previously delineated in the isolated alpha subunit (αTS) in the absence of the beta subunit (βTS). The amino acid interaction networks were different between the ligand-free enzyme and the enzyme actively catalyzing turnover. Previous X-ray crystallography studies indicated only minor localized changes when ligands bind αTS, and so, structural changes alone could not explain the changes to the amino acid interaction networks. We hypothesized that the network changes could instead be related to changes in conformational dynamics. As such, we conducted nuclear magnetic resonance relaxation studies on different substrate- and products-bound complexes of αTS. Specifically, we collected 15N R2 relaxation dispersion data that reports on microsecond-to-millisecond timescale motion of backbone amide groups. These experiments indicated that there are conformational exchange events throughout αTS. Substrate and product binding change specific motional pathways throughout the enzyme, and these pathways connect the previously identified network residues. These pathways reach the αTS/βTS binding interface, suggesting that the identified dynamic networks may also be important for communication with the βTS subunit.


ISOLATION FROM NORMAL MICE OF A PNEUMOTROPIC VIRUS WHICH FORMS ELEMENTARY BODIES.
1. A pneumotropic virus which forms elementary bodies has been isolated from apparently normal albino Swiss mice. 2. The antigenic relationship of this virus to those of meningopneumonitis, lymphogranuloma venereum, hamster pneumonia (7), and human pneumonitis (8) was established either by cross-immunity or complement fixation or both. 3. In spite of a relationship to other viruses, the virus could be differentiated from all the others studied by certain of its properties.


Vasomotor projections from the anteroventral third ventricle (AV3V) region.
The anteroventral third ventricle (AV3V) region is necessary for the development of a variety of models of experimental hypertension. Neuroanatomic studies have identified descending projections from this region to the mesencephalic central gray and tegmentum. This study was done to identify functionally the projections involved in cardiovascular regulation from the AV3V region to the mesencephalon. Regional hemodynamic responses to electrical stimulation of the AV3V region were compared with responses to stimulation of the rostral central gray, and these responses were compared before and after ablation of mesencephalic tissue. Stimulation of the AV3V region elicited hindquarter, vasodilation and renal and mesenteric vasoconstriction associated with a depressor response and bradycardia. These responses were partially dependent on adrenal catecholamines since adrenalectomy reduced hindquarter, mesenteric, and depressor responses. Regional hemodynamic responses to central gray stimulation were qualitatively similar and also demonstrated a partial dependence on adrenal catecholamines. After electrolyte ablation of central gray tissue, significant reductions in hindquarter, mesenteric, and depressor responses to AV3V stimulation were observed. These observations suggest that a cardiovascular control system important for the development of several models of hypertension may be dependent on neuronal projections from the rostral medial hypothalamus through the mesencephalic central gray.


Depressive personality vulnerability in chronic physical pain: Centrality of sociotropy.
Chronic physical pain is one of modern medicine's principal challenges. Recently, there has been a keen research interest in the role of depressive personality vulnerability (DPV) in the course of chronic pain. This is the first attempt to examine the role of three leading DPV dimensions-sociotropy, autonomy, and self-criticism-in chronic pain. Chronic pain patients (N = 428) were assessed four times as to their pain, disability, anxious depression, and pain-based catastrophizing. At Time 1, sociotropy, autonomy, and self-criticism were also assessed. The effects of sociotropy, autonomy, and self-criticism on pain, disability, anxious depression, and pain-based catastrophizing were examined using structural equation modeling analyses. All DPV dimensions uniquely predicted Time 1, but not Time 2, anxious depression. Sociotropy predicted Time 1 pain and catastrophizing over and above anxious depression, as well as an increase in catastrophizing over time. Autonomy predicted a decrease in catastrophizing and disability, and Time 1 anxious depression predicted an increase in self-criticism. Sociotropy appears to be a unique dimension of DPV in chronic pain.


Line Integral Alternating Minimization Algorithm for Dual-Energy X-Ray CT Image Reconstruction.
We propose a new algorithm, called line integral alternating minimization (LIAM), for dual-energy X-ray CT image reconstruction. Instead of obtaining component images by minimizing the discrepancy between the data and the mean estimates, LIAM allows for a tunable discrepancy between the basis material projections and the basis sinograms. A parameter is introduced that controls the size of this discrepancy, and with this parameter the new algorithm can continuously go from a two-step approach to the joint estimation approach. LIAM alternates between iteratively updating the line integrals of the component images and reconstruction of the component images using an image iterative deblurring algorithm. An edge-preserving penalty function can be incorporated in the iterative deblurring step to decrease the roughness in component images. Images from both simulated and experimentally acquired sinograms from a clinical scanner were reconstructed by LIAM while varying the regularization parameters to identify good choices. The results from the dual-energy alternating minimization algorithm applied to the same data were used for comparison. Using a small fraction of the computation time of dual-energy alternating minimization, LIAM achieves better accuracy of the component images in the presence of Poisson noise for simulated data reconstruction and achieves the same level of accuracy for real data reconstruction.


Oral contraceptives and breast cancer: review of the epidemiologic literature.
Oral contraceptives have caused little or no overall increase in risk of breast cancer in women in developed countries, even in long-term users or after more than two decades since initial exposure. Limited evidence suggests a possible small increase in risk in users of oral contraceptives in developing countries, and further studies of breast cancer and oral contraceptives in low risk populations are warranted. Oral contraceptives may enhance risk of breast cancer in young women with a history of benign breast disease, and this possibility should also be investigated further. Multiple investigations have fairly consistently shown risk of breast cancer in women under age 45 years to be increased slightly in long-term users of oral contraceptives. Further studies should be conducted to elucidate the mechanism of this apparent phenomenon.


On the modularity of the intrinsic flexibility of the µ opioid receptor: a computational study.
The µ opioid receptor (µOR), the principal target to control pain, belongs to the G protein-coupled receptors (GPCRs) family, one of the most highlighted protein families due to their importance as therapeutic targets. The conformational flexibility of GPCRs is one of their essential characteristics as they take part in ligand recognition and subsequent activation or inactivation mechanisms. It is assessed that the intrinsic mechanical properties of the µOR, more specifically its particular flexibility behavior, would facilitate the accomplishment of specific biological functions, at least in their first steps, even in the absence of a ligand or any chemical species usually present in its biological environment. The study of the mechanical properties of the µOR would thus bring some indications regarding the highly efficient ability of the µOR to transduce cellular message. We therefore investigate the intrinsic flexibility of the µOR in its apo-form using all-atom Molecular Dynamics simulations at the sub-microsecond time scale. We particularly consider the µOR embedded in a simplified membrane model without specific ions, particular lipids, such as cholesterol moieties, or any other chemical species that could affect the flexibility of the µOR. Our analyses highlighted an important local effect due to the various bendability of the helices resulting in a diversity of shape and volume sizes adopted by the µOR binding site. Such property explains why the µOR can interact with ligands presenting highly diverse structural geometry. By investigating the topology of the µOR binding site, a conformational global effect is depicted: the correlation between the motional modes of the extra- and intracellular parts of µOR on one hand, along with a clear rigidity of the central µOR domain on the other hand. Our results show how the modularity of the µOR flexibility is related to its pre-ability to activate and to present a basal activity.


Is the use of renin-angiotensin system inhibitors in patients with aortic valve stenosis safe and of prognostic benefit? A systematic review and meta-analysis.
Aortic valve stenosis (AVS) is associated with significant morbidity and mortality, especially in the presence of symptoms and echocardiographic signs of left ventricular remodelling (i.e. increase in left ventricular mass, left ventricular dilation, and systolic dysfunction). Renin-angiotensin system inhibitors (RASi) attenuate cardiac remodelling in various conditions, but the safety and efficacy of RASi in AVS is unsure. We performed a systematic review and meta-analysis to address these issues. We identified three smaller randomized clinical trials and five observational studies eligible for inclusion (PubMed, EMBASE, and Cochrane library search criteria: aortic stenosis, aortic valve, angiotensin-converting enzyme inhibitor in different combinations, published in English at any time up to 1 April 2016). Our analyses suggested that use of RASi was safe, with no observed increase in mortality risk [576/3389 patients receiving RASi vs. 1118/4384 controls died; relative risk 0.93 (95% confidence interval 0.78-1.11), P = 0.44]. Use of RASi was also observed to lower the risk of aortic valve replacement (AVR) surgery [67/2913 patients with RASi vs. 154/3666 controls underwent AVR; relative risk 0.68 (95% confidence interval 0.51-0.91), P = 0.01]. In current clinical practice (based on published literature; mainly observational studies), use of RASi appears to be safe in patients with AVS and may reduce the need for AVR, but the evidence is overall weak. Large-scale randomized clinical trials are warranted to address whether prescription of RASi to treatment-naïve patients may prevent disease progression, delay AVR surgery need, and lower the risk of mortality.


Effort-related decision making in humanized COMT mice: Effects of Val158Met polymorphisms and possible implications for negative symptoms in humans.
Catechol-o-methyltransferase (COMT) is an enzyme that metabolizes catecholamines, and is crucial for clearance of dopamine (DA) in prefrontal cortex. Val158Met polymorphism, which causes a valine (Val) to methionine (Met) substitution at codon 158, is reported to be associated with human psychopathologies in some studies. The Val/Val variant of the enzyme results in higher dopamine metabolism, which results in reduced dopamine transmission. Thus, it is important to investigate the relation between Val158Met polymorphisms using rodent models of psychiatric symptoms, including negative symptoms such as motivational dysfunction. In the present study, humanized COMT transgenic mice with two genotype groups (Val/Val (Val) and Met/Met (Met) homozygotes) and wild-type (WT) mice from the S129 background were tested using a touchscreen effort-based choice paradigm. Mice were trained to choose between delivery of a preferred liquid diet that reinforced panel pressing on various fixed ratio (FR) schedules (high-effort alternative), vs. intake of pellets concurrently available in the chamber (low-effort alternative). Panel pressing requirements were controlled by varying the FR levels (FR1, 2, 4, 8, 16) in ascending and descending sequences across weeks of testing. All mice were able to acquire the initial touchscreen operant training, and there was an inverse relationship between the number of reinforcers delivered by panel pressing and pellet intake across different FR levels. There was a significant group x FR level interaction in the ascending limb, with panel presses in the Val group being significantly lower than the WT group in FR1-8, and lower than Met in FR4. These findings indicate that the humanized Val allele in mice modulates FR/pellet-choice performance, as marked by lower levels of panel pressing in the Val group when the ratio requirement was moderately high. These studies may contribute to the understanding of the role of COMT polymorphisms in negative symptoms such as motivational dysfunctions in schizophrenic patients.


Longitudinal increases in blood biomarkers of inflammation or cardiovascular disease and the incidence of venous thromboembolism.
Essentials Inflammatory and cardiac diseases are associated with increased venous thromboembolism (VTE) risk. Our prospective study assessed rise in inflammatory or cardiac biomarkers and VTE risk. A greater 6-year rise in N-terminal natriuretic peptide is associated with increased VTE incidence. Volume overload or impending cardiac disease may contribute to VTE occurrence. Background We previously showed that participants in the population-based Atherosclerosis Risk in Communities (ARIC) cohort with elevated levels of blood biomarkers of inflammation or cardiac disease were at increased risk of venous thromboembolism (VTE). Objective We hypothesized that ARIC participants with larger 6-year increases in the levels of three biomarkers - C-reactive protein (CRP), N-terminal pro-B-type natriuretic peptide (NT-proBNP), and troponin T - would also have an increased subsequent risk of VTE. Methods We measured changes in the levels of these biomarkers in 9844 participants from 1990-1992 to 1996-1998, and then identified VTEs through 2015. Results A greater 6-year rise in the level of NT-proBNP, but not in that of CRP or troponin T, was significantly associated with increased VTE incidence over a median of 17.6 years of follow-up. After adjustment for other VTE risk factors, those whose NT-proBNP level rose from < 100 pg mL-1 to ≥ 100 pg mL-1 had a hazard ratio for VTE of 1.44 (95% confidence interval [CI] 1.15-1.80), as compared with the reference group with an NT-proBNP level of < 100 pg mL-1 at both times. This hazard ratio was slightly higher (1.66, 95% CI 1.19-2.31) during the first 10 years of follow-up, but was attenuated (1.24, 95% CI 0.99-1.56) after adjustment for prevalent and incident coronary heart disease, heart failure, and atrial fibrillation. Conclusions The two most likely explanations for our results are that: (i) an increasing NT-proBNP level reflects increasing subclinical volume overload and potentially increased venous stasis or subclinical PE that had gone unrecognized over time; or (ii) an increasing NT-proBNP level is a risk marker for impending cardiac disease that places patients at risk of VTE.


Epithelial‑mesenchymal transition in chronic rhinosinusitis (CRS) and the prognostic value of α‑SMA in postoperative outcomes of patients with CRS.
Tissue remodeling is the pathological basis of the symptoms encountered in chronic rhinosinusitis (CRS). Epithelial‑mesenchymal transition (EMT) may participate in this process. The present study was designed to investigate the involvement of EMT in CRS. In addition, the prognostic value of the EMT biomarker α‑smooth muscle actin (α‑SMA) was assessed in patients with CRS who underwent endoscopic sinus surgery (ESS). A total of 13 patients with CRS without nasal polyps (CRSsNP), 13 patients with CRS with nasal polyps (CRSwNP) and 13 control subjects were enrolled. The expression of EMT markers was determined in sinonasal specimens by qPCR, western blot and immunofluorescence assays. EMT features were evaluated in primary nasal epithelial cells (NECs) with transforming growth factor (TGF)‑β1 stimulation. The associations were assessed between α‑SMA expression and the clinical features of CRS. Epithelial and mesenchymal markers were overexpressed in the sinonasal specimens of both CRSsNP and CRSwNP patients. Alterations in the expression pattern were more apparent in the CRSsNP patients. Following incubation of primary NECs with TGF‑β1, a mesenchymal shape was acquired. In addition, NECs that co‑expressed α‑SMA and cytokeratin were readily detected and the protein levels of α‑SMA were elevated. In contrast to α‑SMA, the levels of E‑cadherin were decreased. The protein levels of α‑SMA were negatively correlated with endoscopic scores and several postoperative symptoms. In conclusion, partial EMT occurred in patients with CRS, notably in CRSsNP patients. Moreover, primary NECs could undergo EMT following TGF‑β1 treatment in vitro. In addition, α‑SMA could be considered an efficient predictor for postoperative endoscopic and symptomatic outcomes in patients with CRS treated with ESS.


Configuration of Materially Retained Carbon in Our Society: A WIO-MFA-Based Approach for Japan.
To achieve the goals of Paris Agreement, global society is directing much effort in substantially reducing greenhouse gas (GHG) emissions. In addition to energy-related efforts, prevention of carbon release into the atmosphere with carbon capture and storage (CCS) and/or utilization of biomass resources is considered indispensable to achieving the global objective. In this study, considering carbon-containing goods as carbon reservoirs in our society similar to forests and reservoirs enabling CCS, the flow of materially utilized carbon was quantified by input-output-based material flow analysis (IO-MFA). As a result, in 2011, 6.3 Mt-C of petroleum-derived carbon and 7.9 Mt-C of wood-derived carbon were introduced to the Japanese society as end-use products (e.g., automobiles and constructions) in various forms (e.g., plastics and synthetic rubbers). The total amount (14.2 Mt-C) corresponded to 4.1% (52.1 Mt-CO2) of annual CO2 emission in Japan in 2011. Subsequently, by referring to the technology that can treat carbon in the target forms in end-of-life products, the recoverability of carbon as a material has been discussed with respect to each form and end-use of carbon. By numerically showing the necessity and potential of implementing appropriate technologies, this study provides scientific direction for policymakers to establish a quality carbon cycle in our society.


The effect of recurrent practice at home on the acceptability of capillary blood glucose readings. Accuracy of self blood glucose testing.
To judge how reliably patients perform capillary blood glucose testing over time with recurrent practice at home and to assess if a clinic glucose meter is an acceptable alternative to the clinical laboratory for monitoring patient performance. We compared capillary blood glucose readings obtained by patients with their own equipment and the venous blood glucose determinations by the clinical laboratory at three biweekly visits during the initial phase in 40 subjects attending the diabetes clinic at the Veterans' Affairs Medical Center in Phoenix, Arizona. We also compared patient-generated readings using their own equipment and the readings obtained by the clinic glucose meter and strips at five weekly visits during the second phase in 11 subjects who continued further participation. Error grid analysis was used for both comparisons. Capillary blood glucose readings obtained with clinic glucose meter and strips on one hand were correlated with venous blood glucose levels determined by the clinical laboratory. During the initial phase, 30 subjects consistently obtained clinically acceptable comparisons (zone A on the error grid, i.e., within 20% of the laboratory value) or improved over time, 9 subjects showed deterioration, and 1 subject failed to obtain zone A results on any of the visits. Three subjects who had consistently obtained zone A results during the initial phase maintained their performance, whereas eight subjects who had failed to achieve zone A values by the end of the initial phase gradually improved and ultimately achieved zone A values by the end of the study. A highly significant correlation was noted between clinic meter readings and laboratory values (r = 0.93, P < 0.00001). Clinically acceptable user proficiency in capillary blood glucose testing can be maintained in most subjects, with recurrent intensive education during follow-up clinic visits. Therefore, we recommend that these comparisons be performed and patient's technique be observed at each visit to monitor their performance. The clinic glucose meter is a suitable alternative to a clinical laboratory for user proficiency checks.


Private practice dentists' views of oral health injustice.
There is a lack of research examining how dentists in private practice conceptualise the challenge presented by oral health injustice and how attitudes towards low-income patients might be influenced by the commercial nature of private dental care. This study provides insights into how dentists navigated the interactions between the commercial nature of dental practice and the provision of care to patients who either struggled to (or could not) afford the cost of self-funded care in private practice. Participants took part in semi-structured interviews and were invited to keep an online diary of reflections. Thematic analysis was used to examine the data to extract and synthesise understanding of how practitioners conceptualised issues surrounding providing care for the disadvantaged and how this related to the economic realities of private practice. Twenty participants were recruited and interviewed from a variety of private practice environments and roles. This report focuses on one specific theme within the data that explored how participants viewed patients who received public dental care, as well as those privately funded patients who could ill-afford their treatment. The findings raise how neoliberal attitudes towards oral healthcare and dental disease may act as a social determinant of health and contribute to the sustaining of structural barriers and inaction towards oral health injustice. For low-income patients, practitioners distinguish between (a) those who are deemed to be deserving of professional care and the charitable endeavours of the profession, and (b) those who are not. There appears to be no overt professional obligation to actively work towards the amelioration of oral health injustice.


Low-resource method for extracting the malarial biomarker histidine-rich protein II to enhance diagnostic test performance.
We have demonstrated the utility of a self-contained extraction device for the selective isolation, purification, and concentration of the malaria diagnostic protein biomarker Plasmodium falciparum histidine-rich protein II (pfHRPII) from human plasma and whole blood. The extraction cassette consists of a small-diameter tube containing a series of preloaded processing solutions separated by mineral oil valves. Nickel(II) nitrilotriacetic acid-functionalized magnetic particles are added to a parasite-spiked sample contained within the loading chamber of the device for capture of pfHRPII. The biomarker-bound magnetic particles are then entrained by an external magnetic field and transported through three wash solutions. Processing removes sample interfering agents, and the biomarker target is concentrated in the final chamber for subsequent analysis. At parasitemias of 200 parasites/μL, purification and concentration of pfHRPII with extraction efficiencies in excess of 70% total protein target are achieved. The concentration of nonspecific protein interfering agents was reduced by more than 2 orders of magnitude in the final extracted sample without the need for hours of processing time and specialized laboratory equipment. We have demonstrated an application of this low-resource technology by coupling extraction and concentration of pfHRPII within the cassette to a commonly employed rapid diagnostic test. Sample preprocessing improved the visual limit of detection of this test by over 8-fold, suggesting that the combination of both low-resource technologies could prove to be useful in malaria eradication efforts.


Quality assessment of observational studies in psychiatry: an example from perinatal psychiatric research.
In perinatal psychiatry, randomized controlled trials are often not feasible on ethical grounds. Many studies are observational in nature, while others employ large databases not designed primarily for research purposes. Quality assessment of the resulting research is complicated by a lack of standardized tools specifically for this purpose. The aim of this paper is to describe the Systematic Assessment of Quality in Observational Research (SAQOR), a quality assessment tool our team devised for a series of systematic reviews and meta-analyses of evidence-based literature regarding risks and benefits of antidepressant medication during pregnancy.


Ursolic acid alleviates hypercholesterolemia and modulates the gut microbiota in hamsters.
Ursolic acid (UA) is a triterpenoid acid widely abundant in fruits and vegetables such as apple, blueberry and cranberry. The present study was carried out to investigate the effect of UA supplementation in diet on blood cholesterol, intestinal cholesterol absorption and gut microbiota in hypercholesterolemic hamsters. A total of thirty-two hamsters were randomly assigned to four groups and given a non-cholesterol diet (NCD), a high-cholesterol diet containing 0.1% cholesterol (HCD), an HCD diet containing 0.2% UA (UAL), or an HCD diet containing 0.4% UA (UAH) for 6 weeks. Results showed that UA supplementation reduced plasma cholesterol by 15-16% and inhibited intestinal cholesterol absorption by 2.6-9.2%. The in vitro micellar cholesterol solubility experiment clearly demonstrated that UA could displace 40% cholesterol from micelles. In addition, UA decreased the ratio of Firmicutes to Bacteroidetes, whereas it enhanced the growth of short chain fatty acid (SCFA)-producing bacteria in the intestine. In conclusion, UA possessed a cholesterol-lowering activity and could favorably modulate the gut microbiota.