Cell type-specific adaptation of cellular and nuclear volume in micro-engineered 3D environments.
Bio-functionalized three-dimensional (3D) structures fabricated by direct laser writing (DLW) are structurally and mechanically well-defined and ideal for systematically investigating the influence of three-dimensionality and substrate stiffness on cell behavior. Here, we show that different fibroblast-like and epithelial cell lines maintain normal proliferation rates and form functional cell-matrix contacts in DLW-fabricated 3D scaffolds of different mechanics and geometry. Furthermore, the molecular composition of cell-matrix contacts forming in these 3D micro-environments and under conventional 2D culture conditions is identical, based on the analysis of several marker proteins (paxillin, phospho-paxillin, phospho-focal adhesion kinase, vinculin, β1-integrin). However, fibroblast-like and epithelial cells differ markedly in the way they adapt their total cell and nuclear volumes in 3D environments. While fibroblast-like cell lines display significantly increased cell and nuclear volumes in 3D substrates compared to 2D substrates, epithelial cells retain similar cell and nuclear volumes in 2D and 3D environments. Despite differential cell volume regulation between fibroblasts and epithelial cells in 3D environments, the nucleus-to-cell (N/C) volume ratios remain constant for all cell types and culture conditions. Thus, changes in cell and nuclear volume during the transition from 2D to 3D environments are strongly cell type-dependent, but independent of scaffold stiffness, while cells maintain the N/C ratio regardless of culture conditions.


Far-infrared spectroscopy of protein higher-order structures.
Far-infrared (FIR) spectroscopy in the spectral region of 50-450 cm(-1) was used to study a series of protein higher-order structures constructed using β-lactoglobulin and polyomavirus capsid protein VP1. There were marked differences in the spectra for β-lactoglobulin monomer and dimer and between untreated β-lactoglobulin and heat-induced gels formed at neutral pH. Untreated β-lactoglobulin and heat-induced gels formed at acidic pH exhibited little difference in their spectra. Assembly of the quaternary structure of polyomavirus virus-like particles also caused large changes in the FIR spectra. These findings suggest that FIR spectroscopy may prove useful in studying some protein quaternary and higher-order structures. There was evidence of detection of β-lactoglobulin dimerization, intermolecular disulfide bonding in heat-induced neutral gels, and polyomavirus virus-like particle assembly but no evidence that FIR could detect β-lactoglobulin fibrils with their polymeric structure and hydrogen-bonded intermolecular β-pleated sheeting.


Comparison of 3D dose distributions for HDR 192Ir brachytherapy sources with normoxic polymer gel dosimetry and treatment planning system.
Radiation fluence changes caused by the dosimeter itself and poor spatial resolution may lead to lack of 3-dimensional (3D) information depending on the features of the dosimeter and quality assurance of dose distributions for high-dose rate (HDR) iridium-192 ((192)Ir) brachytherapy sources is challenging and experimental dosimetry methods used for brachytherapy sources are limited. In this study, we investigated 3D dose distributions of (192)Ir brachytherapy sources for irradiation with single and multiple dwell positions using a normoxic gel dosimeter and compared them with treatment planning system (TPS) calculations. For dose calibration purposes, 100-mL gel-containing vials were irradiated at predefined doses and then scanned in an magnetic resonance (MR) imaging unit. Gel phantoms prepared in 2 spherical glasses were irradiated with (192)Ir for the calculated dwell positions, and MR scans of the phantoms were obtained. The images were analyzed with MATLAB software. Dose distributions and profiles derived with 1-mm resolution were compared with TPS calculations. Linearity was observed between the delivered dose and the reciprocal of the T2 relaxation time constant of the gel. The x-, y-, and z-axes were defined as the sagittal, coronal, and axial planes, respectively, the sagittal and axial planes were defined parallel to the long axis of the source while the coronal plane was defined horizontally to the long axis of the source. The differences between measured and calculated profile widths of 3-cm source length and point source for 70%, 50%, and 30% isodose lines were evaluated at 3 dose levels using 18 profiles of comparison. The calculations for 3-cm source length revealed a difference of > 3mm in 1 coordinate at 50% profile width on the sagittal plane and 3 coordinates at 70% profile width and 2 coordinates at 50% and 30% profile widths on the axial plane. Calculations on the coronal plane for 3-cm source length showed > 3-mm difference in 1 coordinate at 50% and 70% and 2 coordinates at 30% profile widths. The point source measurements and calculations for 50% profile widths revealed a difference > 3mm in 1 coordinate on the sagittal plane and 2 coordinates on the axial plane. The doses of 3 coordinates on the sagittal plane and 4 coordinates on the axial plane could not be evaluated in 30% profile width because of low doses. There was good agreement between the gel dosimetry and TPS results. Gel dosimetry provides dose distributions in all 3 planes at the same time, which enables us to define the dose distributions in any plane with high resolution. It can be used to obtain 3D dose distributions for HDR (192)Ir brachytherapy sources and 3D dose verification of TPS.


Evidence to Support the Anti-Cancer Effect of Olive Leaf Extract and Future Directions.
The traditional Mediterranean diet (MD) is associated with long life and lower prevalence of cardiovascular disease and cancers. The main components of this diet include high intake of fruit, vegetables, red wine, extra virgin olive oil (EVOO) and fish, low intake of dairy and red meat. Olive oil has gained support as a key effector of health benefits and there is evidence that this relates to the polyphenol content. Olive leaf extract (OLE) contains a higher quantity and variety of polyphenols than those found in EVOO. There are also important structural differences between polyphenols from olive leaf and those from olive fruit that may improve the capacity of OLE to enhance health outcomes. Olive polyphenols have been claimed to play an important protective role in cancer and other inflammation-related diseases. Both inflammatory and cancer cell models have shown that olive leaf polyphenols are anti-inflammatory and protect against DNA damage initiated by free radicals. The various bioactive properties of olive leaf polyphenols are a plausible explanation for the inhibition of progression and development of cancers. The pathways and signaling cascades manipulated include the NF-κB inflammatory response and the oxidative stress response, but the effects of these bioactive components may also result from their action as a phytoestrogen. Due to the similar structure of the olive polyphenols to oestrogens, these have been hypothesized to interact with oestrogen receptors, thereby reducing the prevalence and progression of hormone related cancers. Evidence for the protective effect of olive polyphenols for cancer in humans remains anecdotal and clinical trials are required to substantiate these claims idea. This review aims to amalgamate the current literature regarding bioavailability and mechanisms involved in the potential anti-cancer action of olive leaf polyphenols.


Root-microbe systems: the effect and mode of interaction of Stress Protecting Agent (SPA) Stenotrophomonas rhizophila DSM14405(T.).
Stenotrophomonas rhizophila has great potential for applications in biotechnology and biological control due to its ability to both promote plant growth and protect roots against biotic and a-biotic stresses, yet little is known about the mode of interactions in the root-environment system. We studied mechanisms associated with osmotic stress using transcriptomic and microscopic approaches. In response to salt or root extracts, the transcriptome of S. rhizophila DSM14405(T) changed drastically. We found a notably similar response for several functional gene groups responsible for general stress protection, energy production, and cell motility. However, unique changes in the transcriptome were also observed: the negative regulation of flagella-coding genes together with the up-regulation of the genes responsible for biofilm formation and alginate biosynthesis were identified as a single mechanism of S. rhizophila DSM14405(T) against salt shock. However, production and excretion of glucosylglycerol (GG) were found as a remarkable mechanism for the stress protection of this Stenotrophomonas strain. For S. rhizophila treated with root exudates, the shift from the planktonic lifestyle to a sessile one was measured as expressed in the down-regulation of flagellar-driven motility. These findings fit well with the observed positive regulation of host colonization genes and microscopic images that show different colonization patterns of oilseed rape roots. Spermidine, described as a plant growth regulator, was also newly identified as a protector against stress. Overall, we identified mechanisms of Stenotrophomonas to protect roots against osmotic stress in the environment. In addition to both the changes in life style and energy metabolism, phytohormons, and osmoprotectants were also found to play a key role in stress protection.


Microtensile bond strength of several adhesive systems to different dentin depths.
To determine the bond strength of five adhesives systems to either superficial or deep dentin. Extracted human third molars had their crowns transversally sectioned either next to the occlusal DEJ or next to the pulp, to expose flat, superficial or deep dentin surfaces. The surfaces were bonded with: 1) three two-step, total-etch, self-priming adhesives (Single Bond, Prime&Bond NT, and Excite), 2) a two-step, self-etching primer (Clearfil SE Bond), and 3) a single-step, self-etching all-in-one adhesive (Etch & Prime 3.0) according to manufacturers' directions. Composite build-ups were constructed incrementally with Z250. After storage for 24 hours in water at 37 degrees C, the teeth were sectioned to obtain several bonded beams of 1.0 mm2 cross-sectional area. Each beam was tested in tension in an Instron machine at 0.5 mm/minute. Results were analyzed by two-way ANOVA and post-hoc multiple comparisons (P = 0.05). Bonded interfaces were also examined by TEM. Nanoleakage was examined using a silver-staining technique. Single Bond, Prime&Bond NT and Clearfil SE Bond performed equally when were bonded to superficial dentin; the lowest value was obtained with Etch & Prime 3.0. On deep dentin, the highest bond strengths were attained with Clearfil SE Bond and Prime & Bond NT. The bond strengths of Prime & Bond NT and Excite were significantly higher to deep than to superficial dentin and the rest of the adhesives showed similar bond strength to both dentin depths. Nanoleakage was manifested to variable extent within all hybrid layers examined.


HLA-B27 heavy chains distinguished by a micropolymorphism exhibit differential flexibility.
Although the products of the HLA subtypes B*2705 and B*2709 differ only in residue 116 (Asp versus His) within their peptide-binding grooves, they are differentially associated with inflammatory rheumatic diseases such as ankylosing spondylitis (AS): B*2705 occurs in AS patients, whereas B*2709 is only rarely encountered. The reasons for this distinct association are still unclear but could include subtype-specific conformational and dynamic properties of these antigens. The present study was undertaken to investigate structural and dynamic differences between B*2705 and B*2709 and their possible relationship to subtype-specific disease association. The membrane-distal segments of the B*2705 and B*2709 heavy chains were expressed in vitro and reconstituted together with beta(2)-microglobulin and a peptide. HLA-B27 complexes loaded with 2 self peptides (TIS [RRLPIFSRL] and pVIPR [RRKWRRWHL]) and a sequence-related viral peptide (pLMP2 [RRRWRRLTV]) were studied by isotope-edited infrared spectroscopy to detect differences in their structure and flexibility at physiologic temperature. Our analyses revealed the existence of subtype-specific conformational differences between the 2 HLA-B27 heavy chains at physiologic temperature, which are undetectable using x-ray crystallography. Irrespective of the bound peptide, the heavy chain of the B*2705 complex exhibited higher conformational flexibility than the B*2709 heavy chain. The present study demonstrates the existence of previously undetected systematic conformational and dynamic differences between the heavy chains of the 2 HLA-B27 subtypes. Since effector cell recognition of cells expressing HLA antigens is dependent on the dynamic properties of the interacting cell surface molecules, this HLA-B27 subtype-specific heavy chain flexibility could have a role in the distinct association of HLA-B27 subtypes with spondylarthritides.


Herbal medicines as diuretics: a review of the scientific evidence.
There is increasing interest in the health and wellness benefits of herbs and botanicals. This is with good reason as they might offer a natural safeguard against the development of certain conditions and be a putative treatment for some diseases. One such area may be the lowering of blood pressure in those where it is elevated (i.e., hypertension). One class of clinical medicines used to lower blood pressure are known as diuretics and work by increasing the excretion of urine from the body as well as the amount of sodium in urine. There are a growing number of studies purporting diuretic effects with traditional medicines. The aim of this article was to review these studies and identify which extracts promote diuresis (which we assessed on terms of urine excreted and urinary sodium excretion) and also to identify the research needs in this area. We identified a number of species and genuses reporting diuretic effects. Of these, the most promising, at the present time, are the species Foeniculum vulgare, Fraxinus excelsior, Hibiscus sabdariffa, Petroselinum sativum and Spergularia purpurea, and species from the genuses Cucumis (Cucumis melo and Cucumis trigonus), Equisetum (Equisetum bogotense, Equisetum fluviatile, Equisetum giganteum, Equisetum hiemale var. affine and Equisetum myriochaetum), Lepidium (Lepidium latifolium and Lepidium sativum), Phyllanthus (Phyllanthus amarus, Phyllanthus corcovadensis and Phyllanthus sellowianus) and Sambucus (Sambucus mexicana and Sambucus nigra). However, there the number of studies is limited and we recommend that further studies be conducted to confirm reported effects. Such evidence is needed to provide scientific credence to the folklore use of traditional medicines and even be helpful in the development of future medicines, treatments and treatment guidelines.


Wave-shape-keeping media.
By using the coordinate transformation theory, we propose a media that is used to control the propagation of electromagnetic (EM) waves with the front shape and size being kept and thus to guide the wave energy along one certain direction, whatever the wave shape may be. We obtain the general formula of designing such media and present the typical example in the cylindrical EM wave case. Such media have many potential applications such as super waveguides, high direction antennas, wave-shape adapters, and other types of EM devices. The approaches presented in this paper are also applicable to acoustic waves.


Effect of monophasic pulsed current on heel pain and functional activities caused by plantar fasciitis.
Plantar fasciitis (PF) is a soft tissue disorder considered to be one of the most common causes of inferior heel pain. The aim of this study was to investigate the effect of monophasic pulsed current (MPC) and MPC coupled with plantar fascia-specific stretching exercises (SE) on the treatment of PF. Forty-four participants (22 women and 22 men, with a mean age of 49 years) diagnosed with PF were randomly assigned to receive MPC (n=22) or MPC coupled with plantar fascia-specific SE (n=22). Prior to and after 4 weeks of treatment, participants underwent baseline evaluation; heel pain was evaluated using a visual analogue scale (VAS), heel tenderness threshold was quantified using a handheld pressure algometer (PA), and functional activities level was assessed using the Activities of Daily Living subscale of the Foot and Ankle Ability Measure (ADL/FAAM). Heel pain scores showed a significant reduction in both groups compared to baseline VAS scores (P<0.001). Heel tenderness improved significantly in both groups compared with baseline PA scores (P<0.001). Functional activity level improved significantly in both groups compared with baseline (ADL/FAAM) scores (P<0.001). However, no significant differences existed between the 2 treatment groups in all post-intervention outcome measures. This trial showed that MPC is useful in treating inferior heel symptoms caused by PF.


Randomized controlled trial of 7-Day vs. 14-Day antibiotics for neonatal sepsis.
There are no evidence-based guidelines available regarding the duration of antibiotics in neonatal septicemia. We compared the effectiveness of a 7-day intravenous antibiotic regimen with the standard 14-day regime in blood-culture-proven sepsis in neonates. This was a controlled, blinded, randomized trial with stratification (for birth weight). Blood-culture-positive septic babies > or =32 weeks and/or > or =1500 g were enrolled if meningitis and other deep-seated focal infections were ruled out. Parental consent was obtained. Randomization to either 7-day or 14-day therapy was done on day 7 of antibiotics if the baby had clinically remitted by day 5. Blood culture was repeated 24 h after antibiotic completion. Subjects were observed in the hospital for at least 72 h, and followed-up for 28 days by weekly visits and telephonic contacts. The primary outcome was treatment failure within 28 days defined as a positive blood culture, or clinical signs accompanied by either positive CRP or adjudicated to be a relapse by an expert committee. A total of 120 babies were eligible, 51 were excluded (no consent: 12; non-remission: 39), and 69 were randomized to receive either a 7-day course (n = 34) or a 14-day course (n = 35) of antibiotics. Baselines variables were comparable in the two groups. Primary outcome assessment could be done in 33 cases in either group. There was a trend to greater treatment failures in the 7-day group compared with 14-day group (5 vs. 1, respectively; P = 0.19). On subgroup analysis of subjects with Staphylococcus aureus infection, those who received 7-day therapy (n = 7) had significantly more treatment failure than 14-day therapy (n = 7) (four and zero, respectively; P = 0.022), whereas on sub-group analysis of babies with non-S. aureus infections, treatment failure rates were identical (3.8% in both groups). On comparing the organisms isolated in the group of subjects which was not randomized by virtue of being symptomatic (n = 39) vs. the group which was randomized (n = 69), it was found that S. aureus infections were significantly commoner in the former group (61.5 vs. 21.3%, respectively; P < 0.001). Neonates > or =32 weeks and/or > or =1500 g with S. aureus sepsis require 14 days of antibiotics. S. aureus infection is also associated with failure to achieve clinical remission by the 5th day of antibiotic therapy. Larger trials are required to confirm whether neonates with non-S. aureus sepsis, whose symptoms remit by 5 days, can be treated with 7 days of antibiotics.


Titania coating of mesoporous silica nanoparticles for improved biocompatibility and drug release within blood vessels.
Blood vessel disease is a major contributor to cardiovascular morbidity and mortality and is hallmarked by dysfunction of the lining endothelial cells (ECs). These cells play a significant role in vascular homeostasis, through the release of mediators to control vessel diameter, hence tissue perfusion. Mesoporous silica nanoparticles (MSNs) can be used as potential drug delivery platforms for vasodilator drugs. Here, using an ex vivo model of vascular function, we examine the use of titania coating for improved biocompatibility and release dynamics of MSN loaded sodium nitroprusside (SNP). MSNs (95 ± 23 nm diameter; pore size 2.7 nm) were synthesised and fully characterised. They were loaded with SNP and coated with titania (TiO2), using the magnetron sputtering technique. Pre-constricted aortic vessels were exposed to drug loaded MSNs (at 1.96 × 1012 MSN mL-1) and the time course of vessel dilation observed, in real time. Exposure of viable vessels to MSNs lead to their internalization into the cytoplasm of ECs, while TiMSNs were also observed in the elastic lamina and smooth muscle cell layers. We demonstrate that titania coating of MSNs significantly improves their biocompatibility and alters the dynamics of drug release. A slow and more sustained relaxation was evident after uptake of TiMSN-SNP, in comparison to uncoated MSN-SNP (rate of dilation was 0.08% per min over a 2.5 h period). The use of titania coated MSNs for drug delivery to the vasculature may be an attractive strategy for therapeutic clinical intervention in cardiovascular disease. Cardiovascular disease is a major cause of mortality and morbidity worldwide, with a total global cost of over $918 billion, by 2030. Mesoporous silica nanoparticles (MSNs) have great potential for the delivery of drugs that can treat vessel disease. This paper provides the first description for the use of titania coated MSNs with increased vascular penetration, for the delivery of vasodilator drugs, without compromising overall vessel function. We demonstrate that titania coating of MSNs significantly improves their biocompatibility and uptake within aortic blood vessels and furthermore, enables a slower and more sustained release of the vasodilator drug, sodium nitroprusside within the vessel, thus making them an attractive strategy for the treatment of vascular disease.


Oncogenic effects of WNT5A in Epstein-Barr virus‑associated nasopharyngeal carcinoma.
The molecular events that drive the progression of Epstein-Barr virus (EBV)-associated nasopharyngeal carcinoma (NPC) are still to be elucidated. Here, we report for the first time the pathogenic significance of an NPC-associated gene, wingless-type MMTV integration site family, member 5A (WNT5A) and the contribution of EBV to its expression. WNT5A is a representative Wnt protein that activates non-canonical Wnt signalling. With regard to its role in carcinogenesis, there is conflicting evidence as to whether WNT5A has a tumour-promoting or tumour-suppressive role. We show that WNT5A is upregulated in primary NPC tissue samples. We also demonstrate that WNT5A expression was dramatically increased in NPC cell lines expressing the EBV-encoded LMP2A gene, suggesting that this EBV-encoded latent gene is responsible for upregulating WNT5A in NPC. In addition, in vitro WNT5A overexpression promotes the proliferation, migration and invasion of NPC cells. Our results not only reveal pro-tumorigenic effects of WNT5A in NPC but also suggest that WNT5A could be an important therapeutic target in patients with EBV-associated disease.


Heterogeneity of tardive dyskinesia. A multivariate analysis.
To determine whether tardive dyskinesia (TD) is a single abnormal movement syndrome or multiple syndromes involving different anatomical areas, we examined 228 out-patients diagnosed with TD at the Connecticut Mental Health Center in New Haven. Application of factor analysis to the seven anatomical severity scores of the Abnormal Involuntary Movement Scale yielded three statistically independent factors involving abnormal movements primarily of the jaw-tongue, face-lips, and extremities-trunk. Using logistic regression to predict the severity of these factors, we found that the severity of the orofacial scores was positively associated with age, schizoaffective or affective disorder, and living alone, while severity of non-orofacial movement was positively associated with current neuroleptic dose, non-use of psychiatric medication, and living alone. Our findings suggest that orofacial and non-orofacial dyskinetic movements may involve distinct clinical syndromes of TD, each having a different set of prognostic and, possibly, aetiological determinants.


Compensation algorithm for the phase-shift error of polarization-based parallel two-step phase-shifting digital holography.
We propose an algorithm for compensating the phase-shift error of polarization-based parallel two-step phase-shifting digital holography, which is a technique for recording a spatial two-step phase-shifted hologram. Although a polarization-based system of the technique has been experimentally demonstrated, there had been the problem that the phase difference of two phase-shifted holograms had been changed by the extinction ratio of the micropolarizer array attached to the image sensor used in the system. To improve the performance of the system, we established and formulated an algorithm for compensating the phase-shift error. Accurate spatial phase-shifting interferometry in the system can be conducted by the algorithm regardless of phase-shift error due to the extinction ratio. By the numerical simulation, the proposed algorithm was capable of reducing the root mean square errors of the reconstructed image by 1/4 and 1/5 in amplitude and phase, respectively. Also, the algorithm was experimentally demonstrated, and the experimental results showed that the system employing the proposed algorithm suppressed the conjugate image, which slightly appeared in the image reconstructed by the system not employing the algorithm, even when the extinction ratio was 10:1. Thus, the effectiveness of the proposed algorithm was numerically and experimentally verified.


Development and experimental validation of a finite element model of total ankle replacement.
Total ankle replacement remains a less satisfactory solution compared to other joint replacements. The goal of this study was to develop and validate a finite element model of total ankle replacement, for future testing of hypotheses related to clinical issues. To validate the finite element model, an experimental setup was specifically developed and applied on 8 cadaveric tibias. A non-cemented press fit tibial component of a mobile bearing prosthesis was inserted into the tibias. Two extreme anterior and posterior positions of the mobile bearing insert were considered, as well as a centered one. An axial force of 2kN was applied for each insert position. Strains were measured on the bone surface using digital image correlation. Tibias were CT scanned before implantation, after implantation, and after mechanical tests and removal of the prosthesis. The finite element model replicated the experimental setup. The first CT was used to build the geometry and evaluate the mechanical properties of the tibias. The second CT was used to set the implant position. The third CT was used to assess the bone-implant interface conditions. The coefficient of determination (R-squared) between the measured and predicted strains was 0.91. Predicted bone strains were maximal around the implant keel, especially at the anterior and posterior ends. The finite element model presented here is validated for future tests using more physiological loading conditions.


Epidemiologically and clinically relevant Group B Streptococcus isolates do not bind collagen but display enhanced binding to human fibrinogen.
Group B Streptococcus (GBS) is the leading cause of neonatal septicemia and meningitis. Pili appendages were shown to play a critical role in bacterial adhesion and colonization of human tissues. Recently it was claimed that binding of the pilus-associated adhesin PilA to collagen is a critical, initial step in promoting interactions with the α2β1 integrin expressed on brain endothelial cells. Here we show that strain NCTC10/84 used in this study is not representative for GBS isolates and question the importance of collagen as a critical extracellular matrix component for GBS infections of the central nervous system.


Visualization of early intramembranous ossification by electron microscopic and spectroscopic imaging.
We present electron microscopic and electron spectroscopic images of putative nucleation sites and early mineral deposits during intramembranous ossification of the murine perichondrial ring. Electron spectroscopic imaging (ESI) permits the quantitative determination and direct visualization of spatial distribution of atomic elements within specimens at high spatial resolution. In this study ESI was used to determine the elemental distributions of phosphorus, sulfur, and calcium. Nucleation and subsequent mineralization in the perichondrial ring occurred sequentially along the longitudinal axis. Proximal regions of the ring contained a matrix with only a few nucleation sites that are characterized in conventional electron micrographs as small loci of low-density material in which dense particles are located. Elemental maps of these sites that we obtained by ESI reveal a sulfur-containing matrix in which localized concentrations of phosphorus occur. With further maturation the loci became centers for the genesis of numerous dense rods or crystals. These mineral deposits contained increased concentrations of P, S, and Ca, compared with the surrounding matrix. The appearance of S at nucleation sites and its persistence in developing mineral deposits suggests that a sulfur-containing moiety may serve as a locus within the osteoid matrix to attain high local concentrations of Ca and P, which leads to the controlled local formation of calcium phosphates. Calcification of the perichondrial ring has been found to occur in the absence of matrix vesicles, which illustrates that these membrane-bounded organelles are not obligatory sites for nucleation in this matrix.


Recombinant adeno-associated virus-mediated global anterograde delivery of glial cell line-derived neurotrophic factor to the spinal cord: comparison of rubrospinal and corticospinal tracts in the rat.
Amyotrophic lateral sclerosis (ALS) is characterized by progressive loss of spinal lower motoneurons. Gene delivery is a promising strategy to deliver therapeutic molecules to these vulnerable cells. However, definition of an optimal route of delivery capable of accessing neurons over a considerable extent of the neuraxis represents a significant logistical problem. Intramuscular vector injections are not ideal as this approach would involve hundreds of injections to completely treat an ALS patient and also would be dependent on retrograde transport of the viral platform of choice. Alternatively, upper motoneurons could deliver trophic factors over considerable distances by anterograde transport after a relatively localized intracerebral injection. To test this approach, the present study was designed to compare the corticospinal (CST) and rubrospinal (RST) tracts for their ability to transport recombinant adeno-associated virus serotype 5 (rAAV5)-derived green fluorescent protein (GFP) or glial cell line-derived neurotrophic factor (GDNF) to the spinal cord. Unilateral injections of rAAV5-GFP into the red nucleus (RN) or motor cortex of normal rats produced GFP-positive fibers in the appropriate descending tracts extending to the lumbar spinal cord. For both tracts, GFP-positive axonal projections into the spinal gray matter were consistently observed. GDNF immunohistochemistry demonstrated that confirmed RN injections resulted in GDNF-positive fibers projecting into spinal gray matter as seen in the GFP group. In contrast, confirmed cortical rAAV5-GDNF injections resulted in less evident staining in spinal cord. Spinal cord GDNF levels were elevated at distances up to 72 mm from the injection sites, and confirmed that RST-related GDNF transport to spinal cord surpassed CST-associated delivery.


Linking private, for-profit providers to public sector services for HIV and tuberculosis co-infected patients: A systematic review.
Tuberculosis (TB) is the leading cause of infectious disease deaths worldwide and is the leading cause of death among people with HIV. The World Health Organization (WHO) has called for collaboration between public and private healthcare providers to maximize integration of TB/HIV services and minimize costs. We systematically reviewed published models of public-private sector diagnostic and referral services for TB/HIV co-infected patients. We searched PubMed, the Cochrane Central Register of Controlled Trials, Google Scholar, Science Direct, CINAHL and Web of Science. We included studies that discussed programs that linked private and public providers for TB/HIV concurrent diagnostic and referral services and used Review Manager (Version 5.3, 2015) for meta-analysis. We found 1,218 unduplicated potentially relevant articles and abstracts; three met our eligibility criteria. All three described public-private TB/HIV diagnostic/referral services with varying degrees of integration. In Kenya private practitioners were able to test for both TB and HIV and offer state-subsidized TB medication, but they could not provide state-subsidized antiretroviral therapy (ART) to co-infected patients. In India private practitioners not contractually engaged with the public sector offered TB/HIV services inconsistently and on a subjective basis. Those partnered with the state, however, could test for both TB and HIV and offer state-subsidized medications. In Nigeria some private providers had access to both state-subsidized medications and diagnostic tests; others required patients to pay out-of-pocket for testing and/or treatment. In a meta-analysis of the two quantitative reports, TB patients who sought care in the public sector were almost twice as likely to have been tested for HIV than TB patients who sought care in the private sector (risk ratio [RR] 1.98, 95% confidence interval [CI] 1.88-2.08). However, HIV-infected TB patients who sought care in the public sector were marginally less likely to initiate ART than TB patients who sought care from private providers (RR 0.89, 95% CI 0.78-1.03). These three studies are examples of public-private TB/HIV service delivery and can potentially serve as models for integrated TB/HIV care systems. Successful public-private diagnostic and treatment services can both improve outcomes and decrease costs for patients co-infected with HIV and TB.


Quantifying the incidence of severe-febrile-illness hospital admissions in sub-Saharan Africa.
Severe-febrile-illness (SFI) is a common cause of morbidity and mortality across sub-Saharan Africa (SSA). The burden of SFI in SSA is currently unknown and its estimation is fraught with challenges. This is due to a lack of diagnostic capacity for SFI in SSA, and thus a dearth of baseline data on the underlying etiology of SFI cases and scant SFI-specific causative-agent prevalence data. To highlight the public health significance of SFI in SSA, we developed a Bayesian model to quantify the incidence of SFI hospital admissions in SSA. Our estimates indicate a mean population-weighted SFI-inpatient-admission incidence rate of 18.4 (6.8-31.1, 68% CrI) per 1000 people for the year 2014, across all ages within areas of SSA with stable Plasmodium falciparum transmission. We further estimated a total of 16,200,337 (5,993,249-27,321,779, 68% CrI) SFI hospital admissions. This analysis reveals the significant burden of SFI in hospitals in SSA, but also highlights the paucity of pathogen-specific prevalence and incidence data for SFI in SSA. Future improvements in pathogen-specific diagnostics for causative agents of SFI will increase the abundance of SFI-specific prevalence and incidence data, aid future estimations of SFI burden, and enable clinicians to identify SFI-specific pathogens, administer appropriate treatment and management, and facilitate appropriate antibiotic use.


Sex differences in cocaine use and experiences: a double standard revived?
Women's use of prescription medication exceeds that of men's and yet is not viewed with the alarm and disapproval that accompanies women's lower levels of use of alcohol and illicit drugs. Reports in the media, based on anecdotal accounts, have identified women as a group at particular risk for cocaine addiction and have suggested that their problems with cocaine are greater than men's. After reviewing the scientific literature and analyzing the results of an original research study, this paper argues that there is no evidence that women's cocaine use exceeds that of men's, that women's rates of use are growing faster than men's, or that female cocaine users experience more problems than male cocaine users. Since the deviant image of the female cocaine user is a social construction lacking a factual basis, we conclude that a different standard is being applied to women who use cocaine than to men who use cocaine.


A Case of Primary Postpartum Bleeding due to Vaginal Laceration after Vaginal Delivery: Successful Management with Transcatheter Arterial Embolization.
Postpartum hemorrhage is one of the major causes of maternal mortality. There are medical and surgical options to control the bleeding, some of which can impair future fertility. Transcatheter arterial embolization might be a useful option in the management of intractable postpartum bleeding before the consideration of more invasive and radical methods. In this report, we report a 33-year-old patient who presented with primary postpartum hemorrhage due to vaginal laceration and was eventually treated with transcatheter arterial embolization.


9,10-Dihydroanthracenyl structures: original ligands for the synthesis of polymetallic complexes through selective π-coordination.
N-Phenylsuccinimides and the corresponding pyrrolidines containing 9,10-dihydroanthracenyl skeletons were used for exploring their ability to coordinate RuCp(+) moieties through the aromatic rings. Electronic and steric tuning of ligands led to mono-, bi- and tri-metallic complexes. A full structural study was carried out in order to characterize the ruthenium organometallic compounds, proving the different coordination modes both in solution (NMR) and in solid state (X-ray diffraction). A modelling study (at DFT level) was completed with the aim to understand the selective π-coordination observed for mono- and bi-metallic complexes.


Three-dimensional structure of bovine NADH:ubiquinone oxidoreductase (complex I) at 22 A in ice.
NADH:ubiquinone oxidoreductase (complex I) is the first and largest complex in the electron transport chain of mitochondria. The bovine complex purified from cardiac muscle consists of at least 42 different subunits with a combined molecular mass of about 890 kDa. The three-dimensional structure of the complex was determined at 22 A from single particles embedded in vitrified ice using electron cryo-microscopy. The structure was calculated using a new program to align particles, to correct for the contrast transfer function of the microscope, and to carry out the three-dimensional reconstruction of the complex. The bovine complex has the overall L-shaped appearance found in earlier studies of the closely related complex I from Neurospora crassa, but it differs by having a thin stalk region linking the membrane-bound globular arm with the intrinsic membrane domain. Thus, the stalk which measures about 30 A in diameter is likely to contain part of the electron transfer pathway linking the NADH binding site in the globular arm with the ubiquinone binding site in the membrane domain. The globular domain of bovine complex I is significantly bigger than that of the N. crassa enzyme, suggesting that the apparent additional subunit complexity of the bovine enzyme is associated with the globular part.


Factors influencing women's decision to seek antenatal care in the Andes of Peru.
This qualitative study aimed to assess factors influencing pregnant women's decision to seek or avoid antenatal care (ANC) in the Andes of Peru. Open-ended, semi-structured interviews were conducted with 24 women utilizing ANC (+) and 10 women avoiding ANC (-). Interviews were translated to English from Quechua and Spanish, transcribed, and analyzed using grounded theory. Factors influencing ANC- women included: expecting criticism for having additional children; long ANC wait time and inconvenient hours of operation; and masculine gender of health workers. For ANC+ women, motivating factors included: maximizing positive health outcomes; past negative maternity experiences; pressure from family members; and avoidance of rumored fines or fees associated with ANC non-attendance and in-hospital deliveries, respectively. Both ANC+ and ANC- women were fearful and embarrassed about possible criticism for having additional children and the gender of the health workers, yet they weighed these factors differently. To better understand how rural women make decisions about ANC attendance, it is important to consider the value they place on the factors influencing their decision, and their emotional assessment of such issues.


Size-related effects and the influence of metabolic traits and morphology on swimming performance in fish.
Energy metabolism fuels swimming and other biological processes. We compared the swimming performance and energy metabolism within and across eight freshwater fish species. Using swim tunnel respirometers, we measured the standard metabolic rate (SMR) and maximum metabolic rate (MMR) and calculated the critical swimming speed (U crit). We accounted for body size, metabolic traits, and some morphometric ratios in an effort to understand the extent and underlying causes of variation. Body mass was largely the best predictor of swimming capacity and metabolic traits within species. Moreover, we found that predictive models using total length or SMR, in addition to body mass, significantly increased the explained variation of U crit and MMR in certain fish species. These predictive models also underlined that, once body mass has been accounted for, U crit can be independently affected by total length or MMR. This study exemplifies the utility of multiple regression models to assess within-species variability. At interspecific level, our results showed that variation in U crit can partly be explained by the variation in the interrelated traits of MMR, fineness, and muscle ratios. Among the species studied, bleak Alburnus alburnus performed best in terms of swimming performance and efficiency. By contrast, pumpkinseed Lepomis gibbosus showed very poor swimming performance, but attained lower mass-specific cost of transport (MCOT) than some rheophilic species, possibly reflecting a cost reduction strategy to compensate for hydrodynamic disadvantages. In conclusion, this study provides insight into the key factors influencing the swimming performance of fish at both intra- and interspecific levels.


GILZ, a new target for the transcription factor FoxO3, protects T lymphocytes from interleukin-2 withdrawal-induced apoptosis.
Interleukin-2 (IL-2) withdrawal is a physiologic process inducing cell death in activated T lymphocytes. Glucocorticoid-induced leucine zipper (GILZ) has recently been identified as a protein modulating T-cell receptor activation by repressing various signaling pathways. We report here that IL-2 deprivation leads to expression of GILZ in T lymphocytes. We then characterized the human gilz promoter and showed that FoxO3 (Forkhead box class O3) binding to the Forkhead responsive elements identified in the promoter is necessary for induction of gilz expression upon IL-2 withdrawal. To assess the functional consequences of this induction, we used 2 strategies, GILZ overexpression and GILZ silencing in murine IL-2-dependent CTLL-2 cells. GILZ overexpression protects CTLL-2 cells from IL-2 withdrawal-induced apoptosis, whereas cell death is accelerated in cells unable to express GILZ. Concomitantly, the expression of Bim is inhibited in GILZ-overexpressing cells and enhanced when GILZ expression is impaired. Furthermore, GILZ inhibits FoxO3 transcriptional activity that leads to inhibition of Bim expression but also to down-regulation of GILZ itself. Therefore, GILZ is a transiently expressed protein induced upon IL-2 withdrawal that protects T cells from the onset of apoptosis.


Decomposing the rural-urban gap in factors associated with childhood immunisation in sub-Saharan Africa: evidence from surveys in 23 countries.
About 31 million children in sub-Saharan Africa (SSA) suffer from immunisation preventable diseases yearly and more than half a million children die because of lack of access to immunisation. Immunisation coverage has stagnated at 72% in SSA over the past 6 years. Due to evidence that full immunisation of children may be determined by place of residence, this study aimed at investigating the rural-urban differential in full childhood immunisation in SSA. The data used for this study consisted of 26 241 children pooled from 23 Demographic and Health Surveys conducted between 2010 and 2018 in SSA. We performed a Poisson regression analysis with robust Standard Errors (SEs) to determine the factors associated with full immunisation status for rural and urban children. Likewise, a multivariate decomposition analysis for non-linear response model was used to examine the contribution of the covariates to the observed rural and urban differential in full childhood immunisation. All analyses were performed using Stata software V.15.0 and associations with a p<0.05 were considered statistically significant. More than half of children in urban settings were fully immunised (52.8%) while 59.3% of rural residents were not fully immunised. In all, 76.5% of rural-urban variation in full immunisation was attributable to differences in child and maternal characteristics. Household wealth was an important component contributing to the rural-urban gap. Specifically, richest wealth status substantially accounted for immunisation disparity (35.7%). First and sixth birth orders contributed 7.3% and 14.9%, respectively, towards the disparity while 7.9% of the disparity was attributable to distance to health facility. This study has emphasised the rural-urban disparity in childhood immunisation, with children in the urban settings more likely to complete immunisation. Subregional, national and community-level interventions to obviate this disparity should target children in rural settings, those from poor households and women who have difficulties in accessing healthcare facilities due to distance.


Dynamometric Analysis of Normative Auricular Stiffness and Comparison With Operated Prominent Ears.
Stiffness of the auricular cartilage is the main determining factor for the choice of operative technique of the prominent ear deformity. The aim of this study is to evaluate the stiffness of normal appearing ears objectively and quantitatively, compare the results with the operated prominent ear patients, and present prospective short-term dynamometric evaluation of the operated prominent ear patients. A total of 190 volunteers without ear deformities were recruited and 9 age groups were formed: group (5-9), group (10-14), group (15-19), group (20-24), group (25-29), group (30-34), group (35-39), group (40-49), and group (50+). Total 28 ears (14 patients) with otoplasty were included in the study as group (operated 5-9) and group (operated 10-14). In addition, 3 patients with prominent ear deformity were prospectively followed for dynamometric changes that occur with otoplasty operation. The auriculocephalic angle (ACA) was measured once and auricle to scalp distance was measured at 4 different standardized levels. Ear stiffness was measured on each ear individually at 4 different points over the antihelix using digital computer-aided dynamometry. Each ear was compared in terms of ACA, distance, and dynamometric values. Dynamometric values tend to increase with age, which increase and peak around 35 years of age and declines after 40 years of age. Measurements of the first 2 age groups were statistically different compared with the other groups. Postoperative dynamometric measurements (DNM) of group (operated 5-9) were similar with normative values of group (5-9) and postoperative satisfaction visual analogue scale (VAS) score was 92.8%. Postoperative DNM of group (operated 10-14) were higher compared with normative values of group (10-14) for each different measuring level and the postoperative satisfaction VAS score was 75.3. A total of 3 patients with prominent ears had lower dynamometric values preoperatively; these values approached closer to normative values of their age group postoperatively. Results show that auricular cartilage stiffens and malleability decreases with increased age. This stiffness peaks in the 35-39 age group and declines after 40 years of age. Dynamometric values increase, at all levels, suggesting increased cartilage stiffness is related to age. In the scope of these results, cartilage sparing techniques are more suitable for 5 to 14 years of age and cartilage-cutting techniques are more suitable for older patients.


Temperature-programmed electrospray-differential mobility analysis for characterization of ligated nanoparticles in complex media.
An electrospray-differential mobility analyzer (ES-DMA) was operated with an aerosol flow-mode, temperature-programmed approach to enhance its ability to characterize the particle size distributions (PSDs) of nanoscale particles (NPs) in the presence of adsorbed and free ligands. Titanium dioxide NPs (TiO2-NPs) stabilized by citric acid (CA) or bovine serum albumin (BSA) were utilized as representative systems. Transmission electron microscopy (TEM) and inductively coupled plasma mass spectrometry were used to provide visual information and elemental-based PSDs, respectively. Results show that the interference resulting from electrospray-dried nonvolatile salt residual nanoscale particles (S-NPs) could be effectively reduced using the thermal treatment process: PSDs were accurately measured at temperatures above 200 °C for CA-stabilized TiO2-NPs and above 400 °C for BSA-stabilized TiO2-NPs. Moreover, TEM confirmed the volumetric shrinkage of S-NPs due to thermal treatment and also showed that the primary structure of TiO2-NPs was relatively stable over the temperature range studied (i.e., below 700 °C). Conversely, the shape factor for TiO2-NPs decreased after treatment above 500 °C, possibly due to a change in the secondary (aggregate) structure. S-NPs from BSA-stabilized TiO2-NPs exhibited higher global activation energies toward induced volumetric shrinkage than those of CA-stabilized TiO2-NPs, suggesting that activation energy is dependent on ligand size. This prototype study demonstrates the efficacy of using ES-DMA coupled with thermal treatment for characterizing the physical state of NPs, even in a complex medium (e.g., containing plasma proteins) and in the presence of particle agglomerates induced by interaction with binding ligands.


Lifestyle factors and stroke risk: exercise, alcohol, diet, obesity, smoking, drug use, and stress.
Various lifestyle factors have been associated with increasing the risk of stroke. These include lack of exercise, alcohol, diet, obesity, smoking, drug use, and stress. Guidelines endorsed by the Centers for Disease Control and Prevention and the National Institutes of Health recommend that Americans should exercise for at least 30 minutes of moderately intense physical activity on most, and preferably all, days of the week. Recent epidemiologic studies have shown a U-shaped curve for alcohol consumption and coronary heart disease mortality, with low-to-moderate alcohol consumption associated with lower overall mortality. High daily dietary intake of fat is associated with obesity and may act as an independent risk factor or may affect other stroke risk factors such as hypertension, diabetes, hyperlipidemia, and cardiac disease. Homocysteine is another important dietary component associated with stroke risk, while other dietary stroke risk factors are thought to be mediated through the daily intake of several vitamins and antioxidants. Smoking, especially current smoking, is a crucial and extremely modifiable independent determinant of stroke. Despite the obstacles to the modification of lifestyle factors, health professionals should be encouraged to continue to identify such factors and help improve our ability to prevent stroke.


The Limited Benefit of Fish Consumption on Risk of Hip Fracture among Men in the Community-Based Hordaland Health Study.
Hip fractures have a high prevalence worldwide. Few studies have investigated whether fish consumption is associated with risk of hip fractures. The objective of the present study was to investigate the effect of fish intake on the subsequent risk of a hip fracture because of the low number of studies on this topic. A community-based prospective cohort study of 2865 men and women from Hordaland county in Norway, born between 1925⁻1927 and enrolled in the study in 1997⁻1999. Information on hip fracture cases was extracted from hospital records until 31 December 2009. Baseline information on the intake of fish was obtained from a semi-quantitative food frequency questionnaire. Cox proportional hazard regression models with death as a competing risk were used to evaluate the association of fish intake with risk of hip fracture. During a mean (SD) follow-up time of 9.6 (2.7) years, 226 hip fractures (72 in men, 154 in women) were observed. The mean (SD) fish intake was 48 (25) g/1000 kcal. The association between fish intake and risk of hip fracture was not linear and displayed a threshold, with low intake of fish being associated with an increased risk of hip fracture in men (HR (Hazard Ratio) = 1.84, 95% CI 1.10, 3.08). In this community-based prospective study of men and women, a low intake of fish was associated with the risk of a hip fracture in men.


Generation and analyses of R8L barttin knockin mouse.
Barttin, a gene product of BSND, is one of four genes responsible for Bartter syndrome. Coexpression of barttin with ClC-K chloride channels dramatically induces the expression of ClC-K current via insertion of ClC-K-barttin complexes into plasma membranes. We previously showed that stably expressed R8L barttin, a disease-causing missense mutant, is retained in the endoplasmic reticulum (ER) of Madin-Darby canine kidney (MDCK) cells, with the barttin β-subunit remaining bound to ClC-K α-subunits (Hayama A, Rai T, Sasaki S, Uchida S. Histochem Cell Biol 119: 485-493, 2003). However, transient expression of R8L barttin in MDCK cells was reported to impair ClC-K channel function without affecting its subcellular localization. To investigate the pathogenesis in vivo, we generated a knockin mouse model of Bartter syndrome that carries the R8L mutation. These mice display disease-like phenotypes (hypokalemia, metabolic alkalosis, and decreased NaCl reabsorption in distal tubules) under a low-salt diet. Immunofluorescence and immunoelectron microscopy revealed that the plasma membrane localization of both R8L barttin and the ClC-K channel was impaired in these mice, and transepithelial chloride transport in the thin ascending limb of Henle's loop (tAL) as well as thiazide-sensitive chloride clearance were significantly reduced. This reduction in transepithelial chloride transport in tAL, which is totally dependent on ClC-K1/barttin, correlated well with the reduction in the amount of R8L barttin localized to plasma membranes. These results suggest that the major cause of Bartter syndrome type IV caused by R8L barttin mutation is its aberrant intracellular localization.


End of Life Care for Patients Dying of Stroke: A Comparative Registry Study of Stroke and Cancer.
Although stroke is a significant public health challenge and the need for palliative care has been emphasized for these patients, there is limited data on end-of-life care for patients dying from stroke. To study the end-of-life care during the last week of life for patients who had died of stroke in terms of registered symptom, symptom management, and communication, in comparison with patients who had died of cancer. This study is a retrospective, comparative registry study. A retrospective comparative registry study was performed using data from a Swedish national quality register for end-of-life care based on WHO`s definition of Palliative care. Data from 1626 patients who had died of stroke were compared with data from 1626 patients who had died of cancer. Binary logistic analyses were used to calculate odds ratios, with 95% CI. Compared to patients who was dying of cancer, the patients who was dying of stroke had a significantly higher prevalence of having death rattles registered, but a significantly lower prevalence of, nausea, confusion, dyspnea, anxiety, and pain. In addition, the stroke group had significantly lower odds ratios for health care staff not to know whether all these six symptoms were present or not. Patients who was dying of stroke had significantly lower odds ratio of having informative communication from a physician about the transition to end-of-life care and of their family members being offered bereavement follow-up. The results indicate on differences in end-of-life care between patients dying of stroke and those dying from cancer. To improve the end-of-life care in clinical practice and ensure it has consistent quality, irrespective of diagnosis, education and implementation of palliative care principles are necessary.


Stochastic model utilizing spectral and spatial characteristics.
In remote sensing, because of physical properties of targets, sensor pixels in spatial proximity to one another are class conditionally correlated. Our main objective is to exploit this spatial correlation. Therefore, a two-dimensional causal first order Markov model was used to extract the spatial and spectral information and, based upon it, new object classifiers with improved performance were developed. First, the minimum distance (MT) and the maximum likelihood (ML) object classifiers are discussed. Then, based on the proposed model, these two classifiers are modified, and a linear object classifier is introduced. Finally, experimental results are presented.


Structure of an archaeal TYW1, the enzyme catalyzing the second step of wye-base biosynthesis.
Wye bases are tricyclic bases that are found in archaeal and eukaryotic tRNAs. The most modified wye base, wybutosine, which appears at position 37 (the 3'-adjacent position to the anticodon), is known to be important for translational reading-frame maintenance. Saccharomyces cerevisiae TYW1 catalyzes the tri-ring-formation step in wye-base biosynthesis, with the substrate tRNA bearing N(1)-methylated G37. Here, the crystal structure of the archaeal TYW1 homologue from Pyrococcus horikoshii is reported at 2.2 A resolution. The amino-acid sequence of P. horikoshii TYW1 suggested that it is a radical-AdoMet enzyme and the tertiary structure of P. horikoshii TYW1 indeed shares the modified TIM-barrel structure found in other radical-AdoMet enzymes. Radical-AdoMet enzymes generally contain one or two iron-sulfur (FeS) clusters. The tertiary structure of P. horikoshii TYW1 revealed two FeS cluster sites, each containing three cysteine residues. One FeS cluster site was expected from the amino-acid sequence and the other involves cysteine residues that are dispersed throughout the sequence. The existence of two FeS clusters was confirmed from the anomalous Fourier electron-density map. By superposing the P. horikoshii TYW1 tertiary structure on those of other radical-AdoMet enzymes, the AdoMet molecule, which is necessary for the reactions of radical-AdoMet enzymes, was modelled in P. horikoshii TYW1. Surface plots of conservation rates and electrostatic potentials revealed the highly conserved and positively charged active-site hollow. On the basis of the surface properties, a docking model of P. horikoshii TYW1, the tRNA, the FeS clusters and the AdoMet molecule was constructed, with the nucleoside at position 37 of tRNA flipped out from the canonical tRNA structure.


On the relation between brain potentials and the awareness of voluntary movements.
We investigated the relation between neural events and the perceived time of voluntary actions or the perceived time of initiating those actions using the method of Libet. No differences were found in either movement-related potentials or perceived time of motor events between a fixed movement condition, where subjects made voluntary movements of a single finger in each block, and a free movement condition, in which subjects chose whether to respond with the left or the right index finger on each trial. We next calculated both the readiness potential (RP) and lateralised readiness potential (LRP) for trials with early and late times of awareness. The RP tended to occur later on trials with early awareness of movement initiation than on trials with late awareness, ruling out the RP as a cause of our awareness of movement initiation. However, the LRP occurred significantly earlier on trials with early awareness than on trials with late awareness, suggesting that the processes underlying the LRP may cause our awareness of movement initiation.


T-box genes in the ascidian Ciona intestinalis: characterization of cDNAs and spatial expression.
Members of the T-box family of transcription factors share an evolutionarily conserved DNA-binding domain and play significant roles in various processes of embryonic development. Vertebrate T-box genes are categorized into the following five major subfamilies (eight groups), depending on sequence similarities: Brachyury, Tbx1 (Tbx1/10, Tbx15/18/22, Tbx20), Tbx2/3/4/5 (Tbx2/3 and Tbx4/5), Tbx6, and Tbr/Eomes/TBX21. Ascidians are primitive chordates, and their tadpole larva are considered to represent the simplified and basic body plan of vertebrates. In addition, it has been revealed that the ascidian genome contains the basic ancestral complement of genes involved in development. The present characterization of cDNAs and survey of the Ciona intestinalis draft genome demonstrated that the Ciona genome contains a single copy gene for each of the Brachyury, Tbx1/10, Tbx15/18/22, Tbx20, Tbx2/3, and Tbr/Eomes/TBX21 groups, and at least three copies of the Tbx6 subfamily. Each of the Ciona T-box genes shows a characteristic expression pattern, although that of Tbx20 was not determined in the present study. These results provide basic information that will be useful for future studies of the function of each gene, genetic cascades of different T-box genes, and genome-wide surveys of evolutionary changes in the T-box gene structure and organization in this primitive chordate.


Glycogene expression alterations associated with pancreatic cancer epithelial-mesenchymal transition in complementary model systems.
The ability to selectively detect and target cancer cells that have undergone an epithelial-mesenchymal transition (EMT) may lead to improved methods to treat cancers such as pancreatic cancer. The remodeling of cellular glycosylation previously has been associated with cell differentiation and may represent a valuable class of molecular targets for EMT. As a first step toward investigating the nature of glycosylation alterations in EMT, we characterized the expression of glycan-related genes in three in-vitro model systems that each represented a complementary aspect of pancreatic cancer EMT. These models included: 1) TGFβ-induced EMT, which provided a look at the active transition between states; 2) a panel of 22 pancreatic cancer cell lines, which represented terminal differentiation states of either epithelial-like or mesenchymal-like; and 3) actively-migrating and stationary cells, which provided a look at the mechanism of migration. We analyzed expression data from a list of 587 genes involved in glycosylation (biosynthesis, sugar transport, glycan-binding, etc.) or EMT. Glycogenes were altered at a higher prevalence than all other genes in the first two models (p<0.05 and <0.005, respectively) but not in the migration model. Several functional themes were shared between the induced-EMT model and the cell line panel, including alterations to matrix components and proteoglycans, the sulfation of glycosaminoglycans; mannose receptor family members; initiation of O-glycosylation; and certain forms of sialylation. Protein-level changes were confirmed by Western blot for the mannose receptor MRC2 and the O-glycosylation enzyme GALNT3, and cell-surface sulfation changes were confirmed using Alcian Blue staining. Alterations to glycogenes are a major component of cancer EMT and are characterized by changes to matrix components, the sulfation of GAGs, mannose receptors, O-glycosylation, and specific sialylated structures. These results provide leads for targeting aggressive and drug resistant forms of pancreatic cancer cells.


Gene expression of osteoclast differentiation factor is induced by lipopolysaccharide in mouse osteoblasts via Toll-like receptors.
Osteoclast differentiation factor (ODF), a recently identified cytokine of the TNF family, is expressed as a membrane-associated protein in osteoblasts and stromal cells. ODF stimulates the differentiation of osteoclast precursors into osteoclasts in the presence of M-CSF. Here we investigated the effects of LPS on the gene expression of ODF in mouse osteoblasts and an osteoblast cell line and found that LPS increased the ODF mRNA level. A specific inhibitor of extracellular signal-regulated kinase or protein kinase C inhibited this up-regulation, indicating that extracellular signal-regulated kinase and protein kinase C activation was involved. A protein synthesis inhibitor, cycloheximide, rather enhanced the LPS-mediated increase of ODF mRNA, and both a neutralizing Ab of TNF-alpha and a specific inhibitor of PGE synthesis failed to block the ODF mRNA increase by native LPS. Thus, LPS directly induced ODF mRNA. Mouse osteoblasts and an osteoblast cell line constitutively expressed Toll-like receptor (TLR) 2 and 4, which are known as putative LPS receptors. ODF mRNA increases in response to synthetic lipid A were defective in primary osteoblasts from C3H/HeJ mice that contain a nonfunctional mutation in the TLR4 gene, suggesting that TLR4 plays an essential role in the process. Altogether, our results indicate that ODF gene expression is directly increased in osteoblasts by LPS treatment via TLR, and this pathway may play an important role in the pathogenesis of LPS-mediated bone disorders, such as periodontitis.


Measurement of the nonlinear elasticity of red blood cell membranes.
The membranes of human red blood cells (RBCs) are a composite of a fluid lipid bilayer and a triangular network of semiflexible filaments (spectrin). We perform cellular microrheology using the dynamic membrane fluctuations of the RBCs to extract the elastic moduli of this composite membrane. By applying known osmotic stresses, we measure the changes in the elastic constants under imposed strain and thereby determine the nonlinear elastic properties of the membrane. We find that the elastic nonlinearities of the shear modulus in tensed RBC membranes can be well understood in terms of a simple wormlike chain model. Our results show that the elasticity of the spectrin network can mostly account for the area compression modulus at physiological osmolality, suggesting that the lipid bilayer has significant excess area. As the cell swells, the elastic contribution from the now tensed lipid membrane becomes dominant.


Baroreflex control of coronary conductance in normotensive and renal hypertensive conscious dogs with complete heart block.
1. The autonomic and local control of the circumflex coronary bed evoked through acute rises in aortic pressure (aortic balloon inflation) was analysed in conscious, normotensive and hypertensive dogs in complete heart block with ventricles paced at 100 beats/min. 2. In normotensive dogs there was an initial rise (6-12 s) in circumflex conductance preceding a fall in conductance as aortic pressure increased and was sustained. The initial rise, but not the later fall, in circumflex conductance was partly due to a cholinoreceptor and partly due to alpha-adrenoreceptor mechanisms. Local constrictor mechanisms were responsible for the later fall in conductance. 3. The initial cholinoreceptor component of the rise in circumflex conductance is initiated through arterial baroreflex mechanisms. 4. The initial autonomic mechanisms regulating circumflex conductance appear to be absent in renal hypertension. 5. The absence of the transient autonomic rise in circumflex flow and conductance in response to rapid elevations in aortic pressure in certain forms of hypertension, e.g; during tachyarrhythmias and behavioural disturbances, may result in myocardial ischaemia when it is least appropriate.


Comparison of lyophilized versus liquid modified vaccinia Ankara (MVA) formulations and subcutaneous versus intradermal routes of administration in healthy vaccinia-naïve subjects.
Modified vaccinia Ankara (MVA) is being developed as a safer smallpox vaccine and is being placed in the US Strategic National Stockpile (SNS) as a liquid formulation for subcutaneous (SC) administration at a dose of 1×10(8) TCID50 in a volume of 0.5mL. This study compared the safety and immunogenicity of the standard formulation, dose and route with both a more stable, lyophilized formulation and with an antigen-sparing intradermal (ID) route of administration. 524 subjects were randomized to receive either a full dose of Lyophilized-SC, a full dose of Liquid-SC or 20% (2×10(7) TCID50 in 0.1mL) of a full dose Liquid-ID MVA on Days 0 and 28. Safety and immunogenicity were followed through 180 days post second vaccination. Among the 3 groups, the proportion of subjects with moderate/severe functional local reactions was significantly different (P=0.0013) between the Lyophilized-SC group (30.3%), the Liquid-SC group (13.8%) and Liquid-ID group (22.0%) only after first vaccination; and for moderate/severe measured erythema and/or induration after any vaccination (P=0.0001) between the Lyophilized-SC group (58.2%), the Liquid-SC group (58.1%) and the Liquid-ID group (94.8%) and the reactions lasted longer in the Liquid-ID group. In the ID Group, 36.1% of subjects had mild injection site skin discoloration lasting ≥6 months. After second vaccination Day (42-208), geometric mean of peak neutralization titers were 87.8, 49.5 and 59.5 for the Lyophilized-SC, Liquid-SC and Liquid-ID groups, respectively, and the maximum number of responders based on peak titer in each group was 142/145 (97.9%), 142/149 (95.3%) and 138/146 (94.5%), respectively. At 180 days after the second vaccination, geometric mean neutralization titers declined to 11.7, 10.2 and 10.4 with only 54.3%, 39.2% and 35.2% of subjects remaining seropositive for the Lyophilized-SC, Liquid-SC and Liquid-ID groups, respectively. Both the Lyophilized-SC and Liquid-ID groups were considered non-inferior (primary objective) to the Liquid-SC group. Transitioning to a lyophilized formulation, which has a longer shelf life, will not negatively impact immunogenicity. In a situation where insufficient vaccine is available, ID vaccination could be used, increasing the number of available doses of vaccine in the SNS 5-fold (i.e., from 20 million to 100 million doses).


Socioeconomic status and HRT prescribing: a study of practice-level data in England.
Concerns have been raised that women from deprived backgrounds are less likely to be receiving hormone replacement therapy (HRT) treatment and its benefits, although evidence in support of this is lacking. To investigate general practice HRT prescription trends and their association with markers of socioeconomic deprivation. Cross-sectional study of primary care prescribing data in England in 2018. Practice-level prescribing rate was defined as the number of items of HRT prescribed per 1000 registered female patients aged ≥40 years. The association between Index of Multiple Deprivation (IMD) score and HRT prescribing rate was tested using multivariate Poisson regression, adjusting for practice proportions of obesity, smoking, hypertension, diabetes, coronary heart disease and cerebrovascular disease, and practice list size. The overall prescribing rate of HRT was 29% lower in practices from the most deprived quintile compared with the most affluent (incidence rate ratio [IRR] = 0.71; 95% confidence interval [CI] = 0.68 to 0.73). After adjusting for all cardiovascular disease outcomes and risk factors, the prescribing rate in the most deprived quintile was still 18% lower than in the least deprived quintile (adjusted IRR = 0.82; 95% CI = 0.77 to 0.86). In more deprived practices, there was a significantly higher tendency to prescribe oral HRT than transdermal preparations (P<0.001). This study highlights inequalities associated with HRT prescription. This may reflect a large unmet need in terms of menopause care in areas of deprivation. Further research is needed to identify the factors from patient and GP perspectives that may explain this.


Influence of plasma pretreatment on shear bond strength of self-adhesive resin cements to polyetheretherketone.
The aim of this study is to evaluate the adhesion between PEEK and two self-adhesive resin cements after plasma treatment. Eight hundred sixty-four polyetheretherketone (PEEK) disks were cut and polished to silicon carbide (SIC) P4000. One half of the specimens were randomly selected and pretreated with plasma, whereas the remaining 432 specimens remained untreated. Subsequently, specimens were randomly allocated to four groups (n = 108/group): Visio.link (Bredent), Signum PEEK Bond (Heraeus Kulzer), Ambarino P60 (Creamed), and a control group without additional treatment. Half of the specimens of each group (n = 54) were then cemented with either RelyX Unicem Automix 2 (3 M ESPE) or with Clearfil SA (Kuraray). All specimens were stored in water for 24 h (37 °C). Afterwards, specimens were divided into three groups (n = 18) for different aging levels: (1) no aging (baseline measurement), (2) thermal aging for 5,000 cycles (5/55 °C), and (3) thermal aging for 10,000 cycles (5/55 °C). Thereafter, shear bond strengths (SBS) were measured, and failure types (adhesive, mixed, and cohesive) were assessed. Data were analyzed using descriptive statistics, four- and one-way ANOVA followed by a post hoc Scheffé test (p < 0.05). No adhesion could be established without adhesive pretreatment, irrespectively, whether plasma was applied or not. Also, no bond strength was measured when Ambarino P60 was applied. In contrast, adhesive pretreatment resulted in SBS ranging between 8 and 15 MPa. No significant differences were found between the resin cements used. In general, no cohesive failures were observed. Groups without plasma treatment combined with Visio.link or Signum PEEK Bond showed predominantly mixed failure types. Control groups, plasma treated, or treated using Ambarino P60 groups fractured predominantly adhesively. The use of methyl methacrylate (MMA)-based adhesives allows bonding between PEEK and self-adhesive resin cements. Plasma treatment has no impact on bond to resin cements. PEEK reconstructions can be cemented using self-adhesive resin cements combined with pretreatment with MMA-based adhesives.


Relation between childhood experiences and adults' self-esteem: A sample from Baghdad.
Adverse childhood experiences are associated with significant functional impairments and loss of life in adolescence and adulthood. Literature documents the conversion of traumatic emotional experiences in childhood into psychological disorders later in life. The family is one of the most critical risks and resilient factors for mental health in adolescence and emerging adulthood. To estimate the effect of childhood experiences on self-esteem during adulthood in a sample from Baghdad city. This cross-sectional study was conducted in Baghdad city during the period from January 2013 through to January 2014. Multistage sampling techniques were used to choose 13 primary healthcare centers and eight colleges from three universities in Baghdad. Childhood experiences were measured by applying a modified standardized Adverse Childhood Experiences International Questionnaire (ACE-IQ) form. A total of 1040 subjects were surveyed and 1000 responded giving a response rate of 96.2%. The results revealed that 82.7% of the participants were confident within themselves, 14.9% (149) reported to feel a failure, while 28.3% of subjects expressed feeling useless at times. The score for family bonding is expected to significantly increase the score for self-esteem by a mean of 21.48. University, diploma and higher education are expected to significantly decrease the self-esteem score by a mean of - 6.31 compared to those with less than secondary school education. Parents education show statistically insignificant association with the mean score for self-esteem. The findings of this study give an insight into the essential role of childhood experiences in building self-esteem and adaptation later in their life. National health programs are suggested for intervention targeting early adverse childhood experiences and their consequences.


Preoperative variables predict persistent type 2 endoleak after endovascular aneurysm repair.
Persistent type 2 endoleaks (PT2, present >or=6 months) after endovascular aneurysm repair (EVAR) are associated with adverse outcomes. This study evaluated the preoperative risk factors and natural history of PT2 in order to define a population at high risk. From January 1999 to December 2007, 595 of 832 EVAR patients had long-term computed tomography follow-up and comprised the study cohort. Preoperative anatomic and clinical variables were correlated with PT2 using Cox regression. Composite hazard ratios (HRs) were constructed with clusters of high-risk preoperative variables. Primary end points, including spontaneous resolution, sac enlargement >5 mm, and freedom from reintervention, were evaluated using Kaplan-Meier analysis. There were 136 PT2 patients (23%) with a median follow-up of 34.8 months (range, 6.4-121.2 months). Positive predictive factors included patent inferior mesenteric artery (IMA; HR, 4.00; 95% confidence interval [CI], 1.62-9.90; P = .003), increasing number of patent lumbar arteries (HR, 1.24; 95% CI, 1.10-1.41; P = .0006), increasing age (HR, 1.04; 95% CI, 1.01-1.06; P = .005), and increasing luminal diameter on CT-contrast opacified lumen (HR, 1.03; 95% CI, 1.02-1.05; P = .0001). During follow-up, spontaneous PT2 resolution occurred in 34 patients (25%), sac diameter remained stable in 63 (46%), and rupture occurred in 2 (1.5%). Kaplan-Meier analysis estimated that 35.2% +/- 5.6% (95% CI, 23.8%-46.2%) of PT2 resolve spontaneously at 5 years after the index procedure. Freedom from sac enlargement >5 mm was 54.6% +/- 7.2% (95% CI, 40.6%-69.4%) at 5 years. Fifty-nine reinterventions were performed in 39 patients with PT2. Freedom from reintervention was 67.3% +/- 5.0% (95% CI, 57.0%-77.0%) at 5 years. The combination of a patent IMA and one risk factor of more than six patent lumbar arteries, maximum luminal diameter >30 mm, or age >70 years increased the odds of PT2 approximately ninefold. The combination of a patent IMA and any two risk factors increased the odds of PT2 approximately 18-fold. Several readily identifiable preoperative variables are associated with PT2 whose natural history was benign in but 35% of patients. On the basis of the composite high-risk HRs, there is accordingly a cohort of patients in whom perioperative interventions to preclude PT2 should be considered.


Discovery of a potent inhibitor of the antiapoptotic protein Bcl-xL from NMR and parallel synthesis.
The antiapoptotic proteins Bcl-x(L) and Bcl-2 play key roles in the maintenance of normal cellular homeostasis. However, their overexpression can lead to oncogenic transformation and is responsible for drug resistance in certain types of cancer. This makes Bcl-x(L) and Bcl-2 attractive targets for the development of potential anticancer agents. Here we describe the structure-based discovery of a potent Bcl-x(L) inhibitor directed at a hydrophobic groove on the surface of the protein. This groove represents the binding site for BH3 peptides from proapoptotic Bcl-2 family members such as Bak and Bad. Application of NMR-based screening yielded an initial biaryl acid with an affinity (K(d)) of approximately 300 microM for the protein. Following the classical "SAR by NMR" approach, a second-site ligand was identified that bound proximal to the first-site ligand in the hydrophobic groove. From NMR-based structural studies and parallel synthesis, a potent ligand was obtained, which binds to Bcl-x(L) with an inhibition constant (K(i)) of 36 +/- 2 nM.


Anchored boundary conditions for locally isostatic networks.
Finite pieces of locally isostatic networks have a large number of floppy modes because of missing constraints at the surface. Here we show that by imposing suitable boundary conditions at the surface the network can be rendered effectively isostatic. We refer to these as anchored boundary conditions. An important example is formed by a two-dimensional network of corner sharing triangles, which is the focus of this paper. Another way of rendering such networks isostatic is by adding an external wire along which all unpinned vertices can slide (sliding boundary conditions). This approach also allows for the incorporation of boundaries associated with internal holes and complex sample geometries, which are illustrated with examples. The recent synthesis of bilayers of vitreous silica has provided impetus for this work. Experimental results from the imaging of finite pieces at the atomic level need such boundary conditions, if the observed structure is to be computer refined so that the interior atoms have the perception of being in an infinite isostatic environment.


Building a competent health manager at district level: a grounded theory study from Eastern Uganda.
Health systems in low-income countries are often characterized by poor health outcomes. While many reasons have been advanced to explain the persistently poor outcomes, management of the system has been found to play a key role. According to a WHO framework, the management of health systems is central to its ability to deliver needed health services. In this study, we examined how district managers in a rural setting in Uganda perceived existing approaches to strengthening management so as to provide a pragmatic and synergistic model for improving management capacity building. Twenty-two interviews were conducted with district level administrative and political managers, district level health managers and health facility managers to understand their perceptions and definitions of management and capacity building. Kathy Charmaz's constructive approach to grounded theory informed the data analysis process. An interative, dynamic and complex model with three sub-process of building a competent health manager was developed. A competent manager was understood as one who knew his/her roles, was well informed and was empowered to execute management functions. Professionalizing health managers which was viewed as the foundation, the use of engaging learning approaches as the inside contents and having a supportive work environment the frame of the model were the sub-processes involved in the model. The sub-processes were interconnected although the respondents agreed that having a supportive work environment was more time and effort intensive relative to the other two sub-processes. The model developed in our study makes four central contributions to enhance the WHO framework and the existing literature. First, it emphasizes management capacity building as an iterative, dynamic and complex process rather than a set of characteristics of competent managers. Second, our model suggests the need for professionalization of health managers at different levels of the health system. Third, our model underscores the benefits that could be accrued from the use of engaging learning approaches through prolonged and sustained processes that act in synergy. Lastly, our model postulates that different resource investments and a varied range of stakeholders could be required at each of the sub-processes.


Comparison of risk scores for the prediction of stroke in African Americans: Findings from the Jackson Heart Study.
Evidence from existing cohort studies supports the prediction of incident coronary heart disease and stroke using 10-year cardiovascular disease (CVD) risk scores and the American Heart Association/American Stroke Association's cardiovascular health (CVH) metric. We included all Jackson Heart Study participants with complete scoring information at the baseline study visit (2000-2004) who had no history of stroke (n = 4,140). We used Kaplan-Meier methods to calculate the cumulative incidence of stroke and used Cox models to estimate hazard ratios and 95% CIs for stroke according to CVD risk and CVH score. We compared the discrimination of the 2 models according to the Harrell c index and plotted predicted vs observed stroke risk calibration plots for each of the 2 models. The median age of the African American participants was 54.5 years, and 65% were female. The cumulative incidence of stroke increased across worsening categories of CVD risk and CVH. A 1-unit increase in CVD risk increased the hazard of stroke (1.07, 1.06-1.08), whereas each 1-unit increase in CVH corresponded to a decreased hazard of stroke (0.76, 0.69-0.83). As evidenced by the c statistics, the CVH model was less discriminating than the CVD risk model (0.59 [0.55-0.64] vs 0.79 [0.76-0.83]). Both scores were associated with incident stroke in a dose-response fashion; however, the CVD risk model was more discriminating than the CVH model. The CVH score may still be preferable for its simplicity in application to broad patient populations and public health efforts.


Assessment of in vitro effects of direct thrombin inhibitors and activated factor X inhibitors through clot waveform analysis.
Clot waveform analysis (CWA) has been reported to extend the interpretation of clotting time measurement. The parameters obtained from successive derivatives of the clotting reaction curves reflect the rates of activation of individual coagulation factors, theoretically dissecting the cascade pathway. This study aims to assess the in vitro effects of direct thrombin inhibitors (DTIs) and activated factor X (FXa) inhibitors. CWA was applied to the activated partial thromboplastin time (APTT) assay of plasma samples spiked with each drug. For CWA of APTT measurement curves (APTT-CWA), the positive mode of clotting reaction curves was defined as the direction towards fibrin generation. All the maximum positive values in the successive derivatives were decreased dependently on the concentrations of each drug. Moreover, the negative values in the second and third derivatives appeared putatively due to consumption of thrombin and factor FXa, respectively, to form complexes with plasma serine protease inhibitors. The decrease of the maximum negative values observed dependently on the concentrations of each drug appeared to be consistent with the decreased generation of thrombin and factor FXa. The analysis of Hill coefficients of each drug in the dose-response of changes in the APTT-CWA parameters revealed a difference in anticoagulant cooperativity between DTIs versus FXa inhibitors. The APTT-CWA demonstrated evidence for the blockade of thrombin-positive feedback by DTIs and FXa inhibitors and that for the differences in anticoagulant cooperativity between them. The results demonstrate the usability of CWA for assessment of anticoagulation and provide insights into direct anticoagulants.


An imaging comparison of 64Cu-ATSM and 60Cu-ATSM in cancer of the uterine cervix.
Tumor uptake of copper(II)-diacetyl-bis(N(4)-methylthiosemicarbazone) (copper-ATSM), a hypoxia-targeting radiopharmaceutical, assessed by PET has been found to correlate with prognosis in several human cancers. Wide clinical utility of this tracer will require its labeling with a copper radionuclide having a longer half-life than the (60)Cu used in studies to date. The purpose of this work was to obtain the requisite preclinical data for copper-ATSM to file an investigational new drug application, followed by a crossover comparison of PET image quality and tumor uptake with (60)Cu-ATSM and (64)Cu-ATSM in women with cancer of the uterine cervix. The preclinical toxicology and pharmacology of a copper-ATSM formulation was examined using standard in vitro and in vivo assays, as well as 14-d toxicity studies in both rats and rabbits. For the clinical test-retest imaging study, 10 patients with cervical carcinoma underwent PET on separate days with (60)Cu-ATSM and (64)Cu-ATSM. Image quality was assessed qualitatively, and the tumor-to-muscle activity ratio was measured for each tracer. The toxicology and pharmacology data demonstrated that the formulation has an appropriate margin of safety for clinical use. In the patient study, we found that the image quality with (64)Cu-ATSM was better than that with (60)Cu-ATSM because of lower noise. In addition, we found that the pattern and magnitude of tumor uptake of (60)Cu-ATSM and (64)Cu-ATSM on studies separated by 1-9 d were similar. (64)Cu-ATSM appears to be a safe radiopharmaceutical that can be used to obtain high-quality images of tumor hypoxia in human cancers.


Driving forces for the growth of MIB-producing Planktothricoides raciborskii in a low-latitude reservoir.
In comparison with the middle- and high-latitude regions, the low-latitude regions are less associated with the occurrence of 2-methylisoborneol (MIB) episodes, since most of the previously identified MIB producers favor moderate/low light/temperature conditions. Here, we report a serious MIB outbreak over the period from Jul. 2018 to Jun. 2019 in a low-latitude reservoir with a mean annual water temperature of 25.6 °C. The MIB episode lasted for a long period, from Jul. 2018 to Jan. 2019, and Planktothricoides raciborskii was confirmed to be the main MIB producer. The growth characteristics of P. raciborskii were explored through both laboratory culturing and on-site verification experiments. The results indicated that this strain was not nutrient-sensitive at TN > 800 μg L-1 and TP > 10 μg L-1, but favored moderate light intensity (54 μmol photon m-2·s-1) and high temperature (30 °C). The two bloom-forming genera, Limnothrix and Aphanizomenon, favoring lower temperature and similar or relatively higher light intensity, showed much greater proliferation, about 13 folds (Limnothrix) and 58 folds (Aphanizomenon), from Dec. to Jun.; by contrast, the high water temperature (29.9 ± 2.8 °C) and light intensity (189.1 ± 87.6 μmol photon m-2·s-1) from Jul. to Nov. were not favorable to Limnothrix or Aphanizomenon, which might have created an opportunity for the growth of MIB-producing P. raciborskii. In addition, we also found that high temperature could promote the release of MIB from P. raciborskii cells, therefore exerting increased pressure on drinking water treatment processes.


Food insecurity is associated with greater acute care utilization among HIV-infected homeless and marginally housed individuals in San Francisco.
Food insecurity, or the uncertain availability of nutritionally adequate, safe foods, has been associated with poor HIV outcomes. There are few data on the extent to which food insecurity impacts patterns of health-care utilization among HIV-infected individuals. We examined whether food insecurity was associated with hospitalizations, Emergency Department (ED) visits, and non-ED outpatient visits. HIV-infected, homeless and marginally housed individuals participating in the San Francisco Research on Access to Care in the Homeless (REACH) cohort underwent quarterly structured interviews and blood draws. We measured food insecurity with the validated Household Food Insecurity Access Scale, and categorized participants as food secure, mild/moderately food insecure, and severely food insecure. Primary outcomes were: (1) any hospitalizations, (2) any ED visits, and (3) any non-ED outpatient visits. Generalized estimating equations were used to estimate model parameters, adjusting for socio-demographic (age, sex, ethnicity, education, income, housing status, health insurance) and clinical variables (CD4 nadir, time on antiretroviral therapy, depression, and illicit drug use). Beginning in November 2007, 347 persons were followed for a median of 2 years. Fifty-six percent of participants were food insecure at enrollment. Compared with food-secure persons, those with severe food insecurity had increased odds of hospitalizations [adjusted odds ratio (AOR) = 2.16, 95 % confidence interval (CI) = 1.50-3.09] and ED visits (AOR = 1.71, 95 % CI = 1.06-2.30). While the odds of an outpatient visit were 41 % higher for severely food insecure individuals, the effect was not statistically significant (AOR = 1.41, 95 % CI = 0.99-2.01). Mild/moderate food insecurity was also associated with increased hospitalizations (AOR = 1.56, 95 % CI = 1.06-2.30), ED visits (AOR = 1.57, 95 % CI = 1.22-2.03), and outpatient visits (AOR = 1.68, 95 % CI = 1.20-2.17). Food insecurity is associated with increased health services utilization among homeless and marginally housed HIV-infected individuals in San Francisco. Increased ED visits and hospitalizations are not related to fewer ambulatory care visits among food-insecure individuals. Addressing food insecurity should be a critical component of HIV treatment programs and may reduce reliance on acute care utilization.


Involvement of Oxidative and Endoplasmic Reticulum Stress in RDH12-Related Retinopathies.
Retinol dehydrogenase 12 (RDH12) is expressed in photoreceptor inner segments and catalyses the reduction of all-trans retinal (atRAL) to all-trans retinol (atROL), as part of the visual cycle. Mutations in RDH12 are primarily associated with autosomal recessive Leber congenital amaurosis. To further our understanding of the disease mechanisms, HEK-293 cell lines expressing wildtype (WT) and mutant RDH12 were created. The WT cells afforded protection from atRAL-induced toxicity and oxidative stress. Mutant RDH12 cells displayed reduced protein expression and activity, with an inability to protect cells from atRAL toxicity, inducing oxidative and endoplasmic reticulum (ER) stress, with upregulation of sXBP1, CHOP, and ATF4. Pregabalin, a retinal scavenger, attenuated atRAL-induced ER stress in the mutant RDH12 cell lines. A zebrafish rdh12 mutant model (rdh12u533 c.17_23del; p.(Val6AlafsTer5)) was generated through CRISPR-Cas9 gene editing. Mutant fish showed disrupted phagocytosis through transmission electron microscopy, with increased phagosome size at 12 months post-fertilisation. Rhodopsin mislocalisation and reduced expression of atg12 and sod2 indicated early signs of a rod-predominant degeneration. A lack of functional RDH12 results in ER and oxidative stress representing key pathways to be targeted for potential therapeutics.


Clopidogrel with or without omeprazole in coronary artery disease.
Gastrointestinal complications are an important problem of antithrombotic therapy. Proton-pump inhibitors (PPIs) are believed to decrease the risk of such complications, though no randomized trial has proved this in patients receiving dual antiplatelet therapy. Recently, concerns have been raised about the potential for PPIs to blunt the efficacy of clopidogrel. We randomly assigned patients with an indication for dual antiplatelet therapy to receive clopidogrel in combination with either omeprazole or placebo, in addition to aspirin. The primary gastrointestinal end point was a composite of overt or occult bleeding, symptomatic gastroduodenal ulcers or erosions, obstruction, or perforation. The primary cardiovascular end point was a composite of death from cardiovascular causes, nonfatal myocardial infarction, revascularization, or stroke. The trial was terminated prematurely when the sponsor lost financing. We planned to enroll about 5000 patients; a total of 3873 were randomly assigned and 3761 were included in analyses. In all, 51 patients had a gastrointestinal event; the event rate was 1.1% with omeprazole and 2.9% with placebo at 180 days (hazard ratio with omeprazole, 0.34, 95% confidence interval [CI], 0.18 to 0.63; P<0.001). The rate of overt upper gastrointestinal bleeding was also reduced with omeprazole as compared with placebo (hazard ratio, 0.13; 95% CI, 0.03 to 0.56; P = 0.001). A total of 109 patients had a cardiovascular event, with event rates of 4.9% with omeprazole and 5.7% with placebo (hazard ratio with omeprazole, 0.99; 95% CI, 0.68 to 1.44; P = 0.96); high-risk subgroups did not show significant heterogeneity. The two groups did not differ significantly in the rate of serious adverse events, though the risk of diarrhea was increased with omeprazole. Among patients receiving aspirin and clopidogrel, prophylactic use of a PPI reduced the rate of upper gastrointestinal bleeding. There was no apparent cardiovascular interaction between clopidogrel and omeprazole, but our results do not rule out a clinically meaningful difference in cardiovascular events due to use of a PPI. (Funded by Cogentus Pharmaceuticals; ClinicalTrials.gov number, NCT00557921.).