Applying Precision Medicine and Immunotherapy Advances from Oncology to Host-Directed Therapies for Infectious Diseases.
To meet the challenges of increasing antimicrobial resistance, the infectious disease community needs innovative therapeutics. Precision medicine and immunotherapies are transforming cancer therapeutics by targeting the regulatory signaling pathways that are involved not only in malignancies but also in the metabolic and immunologic function of the tumor microenvironment. Infectious diseases target many of the same regulatory pathways as they modulate host metabolic functions for their own nutritional requirements and to impede host immunity. These similarities and the advances made in precision medicine and immuno-oncology that are relevant for the current development of host-directed therapies (HDTs) to treat infectious diseases are discussed. To harness this potential, improvements in drug screening methods and development of assays that utilize the research tools including high throughput multiplexes already developed by oncology are essential. A multidisciplinary approach that brings together immunologists, infectious disease specialists, and oncologists will be necessary to fully develop the potential of HDTs.


Intelligent biogenic pH-sensitive and amine-responsive color-changing label for real-time monitoring of shrimp freshness.
This study developed an intelligent, pH-sensitive and amine-responsive colorimetric label based on chitosan, whey protein and thymol blue by controlling the pH value of the film-forming solution. The obtained label was used to monitor shrimp freshness in real time. The results of this study offer a new approach for developing highly intelligent biogenic labels for freshness monitoring during seafood preservation and processing. The pH 2.0 chitosan-whey protein-thymol blue (CWT-pH 2.0) label exhibited remarkable properties, including the highest tensile strength (5.90 MPa), excellent thermal stability, low water solubility (27.80%) and highly sensitive color responsiveness. The characterization techniques of scanning electron microscopy, X-ray diffraction and Fourier transform infrared spectroscopy confirmed the effective immobilization of thymol blue within the film-forming matrix through hydrogen bonding. Furthermore, the CWT-pH 2.0 label demonstrated visible color changes in the presence of volatile ammonia concentrations ranging from 25 to 25 000 ppm. Consequently, the label successfully facilitated real-time monitoring of shrimp freshness during storage at 4 °C. Importantly, the release rate of thymol blue from the label in food simulants was minimal, measuring only 2.53%. The CWT-pH 2.0 label exhibits significant potential as a highly intelligent biogenic label for freshness monitoring in seafood preservation and processing. © 2023 Society of Chemical Industry.


Patient and Caregiver Perspectives on Terms Used to Describe Kidney Health.
The language used to communicate important aspects of kidney health is inconsistent and may be conceptualized differently by patients and health professionals. These problems may impair the quality of communication, care, and patient outcomes. We aimed to describe the perspectives of patients on terms used to describe kidney health. Patients with CKD (n=54) and caregivers (n=13) from the United States, United Kingdom, and Australia participated in ten focus groups to discuss terms for kidney health (including kidney, renal, CKD, ESKD, kidney failure, and descriptors for kidney function). We analyzed the data using thematic analysis. We identified four themes: provoking and exacerbating undue trauma (fear of the unknown, denoting impending death, despair in having incurable or untreatable disease, premature labeling and assumptions, judgment, stigma, and failure of self); frustrated by ambiguity (confused by medicalized language, lacking personal relevance, baffled by imprecision in meaning, and/or opposed to obsolete terms); making sense of the prognostic enigma (conceptualizing level of kidney function, correlating with symptoms and effect on life, predicting progression, and need for intervention); and mobilizing self-management (confronting reality, enabling planning and preparation, taking ownership for change, learning medical terms for self-advocacy, and educating others). The obscurity and imprecision of terms in CKD can be unduly distressing and traumatizing for patients, which can impair decision making and self-management. Consistent and meaningful patient-centered terminology may improve patient autonomy, satisfaction, and outcomes.


Toll-like receptor-9 expression is inversely correlated with estrogen receptor status in breast cancer.
Toll-like receptor 9 (TLR9) recognizes microbial and vertebrate DNA. We previously demonstrated TLR9 expression in human breast cancer cell lines and showed that TLR9 ligands stimulate their in vitro invasion. The aim of this study was to characterize TLR9 expression in clinical breast cancer specimens. Immunohistochemical staining intensity was compared with known baseline prognostic factors and distant metastasis-free survival. TLR9 expression was detected in 98% of the tumors studied (n = 141). The mean TLR9 staining intensity was higher in ER- than in the highly ER+ breast cancers (p = 0.039). High-grade tumors had significantly increased TLR9 expression (p = 0.027) compared with lower-grade tumors. The highest TLR9 expression was detected in the mucinous and the lowest in the tubular breast cancers (p = 0.034). Distant metastasis-free survival was higher in the lower TLR9-expressing half of the cohort than in the higher TLR9-expressing group (p = 0.118). TLR9 expression did not correlate with menopausal, PgR or Her2 status, patient age, tumor proliferative or invasive characteristics.


Kinetic theory of Jeans instability.
Kinetic treatment of the Jeans gravitational instability, with collisions taken into account, is presented. The initial-value problem for the distribution function which obeys the kinetic equation, with the collision integral conserving the number of particles, is solved. Dispersion relation is obtained and analyzed. New modes are found. Collisions are shown not to affect the Jeans instability criterion. Although the instability growth rate diminishes, the collisions they cannot quench the instability. However, the oscillation spectrum is modified significantly: even in the neighborhood of the threshold frequency omega=0 (separating stable and unstable modes) the spectrum of oscillations can strongly depend on the collision frequency. Propagating (rather than aperiodic) modes are also found. These modes, however, are strongly damped.


Two-year clinical evaluation of a packable resin-based composite.
Packable resin-based composites were introduced in 1998, but few clinical studies have been conducted to evaluate them. The authors conducted a clinical study to determine the two-year performance of SureFil (Dentsply DeTrey GmbH, Konstanz, Germany) packable posterior resin-based composite in Class I and II restorations. An operator (L.S.T.) restored 55 cavities in 36 patients (16 Class I restorations and 39 Class II restorations). After cavity preparation, she etched the enamel with 34 percent phosphoric acid, applied Prime & Bond NT (Dentsply DeTrey GmbH) to dentin and etched enamel for 20 seconds and then cured it for 20 seconds. She restored the cavity using 3- to 5-millimeter increments of SureFil. Independent examiners assessed the restorations after placement and at six months, one year and two years for color matching, marginal discoloration, marginal adaptation, secondary caries, surface texture, anatomical form and postoperative sensitivity, using the Ryge criteria. The authors assessed the changes in the parameters during the two-year period using a software program with Friedman test analysis with a Bonferroni adjustment at significant level of P = .05. At baseline, 31 restorations were graded as Bravo for color match. At the six-month and one-year recall periods (n = 55), 53 restorations remained unchanged. Two restorations from the same patient fell out after one month. After two years (n = 50), there were five Bravos for surface staining and three for marginal adaptation (P < .05). After two years of clinical service, SureFil packable resin-based composite had a success rate of 96 percent, and the authors considered it successful in Class I and II restorations. SureFil packable resin-based composite can be successful in clinical situations with limited-sized cavities and proper application of restorative techniques.


From Burnout to Wellness: A Professional Imperative.
Professional burnout is a barrier to physician wellness. Burnout is prevalent across medicine, and otolaryngology as a specialty ranks near the mean. We review burnout levels in various subgroups of otolaryngology, including academic chairs, faculty, and residents. Risk factors of burnout are discussed, which differ by subgroup. Finally, we propose measures that could help minimize burnout and promote healthy and satisfying careers.


MRNA expression and immunocytochemical localization of leptin receptor in the oviduct of the laying hen (Gallus domesticus).
The role of leptin in female reproduction is fairly well established in mammals, whereas reports concerning leptin action in birds are scarce. The aim of the present study was to detect leptin receptor (LEP-R) mRNA and to localize the leptin receptor protein in the oviduct of laying hens 2h after ovulation by the RT-PCR method and immunocytochemical staining. The RT-PCR reaction demonstrated expression of the long form ofleptin receptor mRNA in all examined oviductal parts (infundibulum, magnum, isthmus and shell gland) and the weakest level was found in the isthmus. The expression of the short isoform was lower than the long form in all examined tissue samples and no differences between oviductal parts were observed. Immunostaining specific for leptin receptor was found in the walls of all examined oviductal parts. The intensity of the immunopositive reaction was the strongest in the epithelium of all examined parts of the oviduct and in the endothelium and muscles of blood vessels. The weakest immunopositive reaction was observed in tubular glands, the connective tissue layer and in circular and longitudinal muscles. The results obtained in this experiment suggest that the oviduct may be a target tissue for leptin, where this polypeptide hormon may participate in egg formation and/or its transport through the oviduct of the domestic hen.


Distinct domains of the GATA-1 cofactor FOG-1 differentially influence erythroid versus megakaryocytic maturation.
FOG family zinc finger proteins play essential roles in development through physical interaction with GATA factors. FOG-1, like its interacting partner GATA-1, is required for normal differentiation of erythroid and megakaryocytic cells. Here, we have developed a functional assay for FOG-1 based on its ability to rescue erythroid and megakaryocytic maturation of a genetically engineered FOG-1(-/-) cell line. We demonstrate that interaction through only one of FOG-1's four GATA-binding zinc fingers is sufficient for rescue, providing evidence against a model in which FOG-1 acts to bridge multiple GATA-binding DNA elements. Importantly, we find that distinct regions of FOG-1 differentially influence erythroid versus megakaryocyte maturation. As such, we propose that FOG-1 may modulate the fate of a bipotential erythroid/megakaryocytic precursor cell.


One-electron electron-molecule potentials consistent with ab initio Møller-Plesset theory.
Ab initio electronic structure methods such as Møller-Plesset (MP) theory can be used to compute electron affinities (EAs) of molecules or clusters of molecules to reasonable accuracy (+/-a few tenths of an electron volt). For systems in which an electron is bound to a closed-shell neutral in a manner that primarily localizes the excess electron exterior to most of the electron density of the neutral, we investigate how a given level of ab initio description can be accurately described by a one-electron potential governing the excess electron's interaction with the neutral. We show what ingredients such a potential must possess not only to reproduce the ab initio EA but also to have long-range electrostatic, polarization, and other contributions identical to the ab initio potential. In particular, we show that using Hartree-Fock level electrostatic moments and polarizability can produce a one-electron potential consistent with MP2 theory. To be consistent with MP3 theory, MP2-level electrostatics and polarizabilities must be used. The long-range components of the ab initio potential are shown to embody both orbital relaxation induced by the excess electron and the dispersion interactions between the excess electron and the other electrons of the neutral. Even though these individual contributions do not necessarily scale as r(-4), they are shown to combine into a total potential that can be represented in the familiar polarization form -(1)/(2)alphar(-4). These findings suggest that electrostatic potentials combined with polarization potentials scaling as r(-4) can indeed describe the relaxation (induction) and dispersion energies of an excess electron. Finally, how these observations might assist in constructing new electron-molecule potentials is also discussed.


Induction of G2 arrest and apoptosis of raji cells by continuous low-dose beta irradiation with 188Re-perrhenate.
Continuous irradiation with exponentially reducing beta-rays induces cell death, known as apoptosis. The aim of this study was to investigate the G2 arrest and apoptosis caused by the beta-ray emitted by the radioisotope (188)Re. Doses of 0.4 Gy (3.7 MBq), 4 Gy (37 MBq), and 40 Gy (370 MBq), were added to Blymphoma Raji cells, and cell viability, apoptosis, and DNA cell-cycle changes were assayed. (188)Re showed time- and dose-dependent effects on cell viability and on cell apoptosis and necrosis. At a (188)Re dose of 0.4 Gy, G(2) cell-cycle arrest was observed after 16 hours, and 4,6-diamidino-2-phenylindole (DAPI) staining indicated a slow, time-dependent increase in apoptotic bodies. At a (188)Re dose of 40 Gy, DNA fragmentation was observed at 2 hours, indicative of early damage in the nucleus. In summary, our results showed that continuous irradiation with low-dose beta-rays induced G(2) arrest and progressive apoptosis, which may be characteristic mechanisms of radionuclide therapy.


Efficient mRNA delivery with graphene oxide-polyethylenimine for generation of footprint-free human induced pluripotent stem cells.
Clinical applications of induced pluripotent stem cells (iPSCs) require development of technologies for the production of "footprint-free" (gene integration-free) iPSCs, which avoid the potential risk of insertional mutagenesis in humans. Previously, several studies have shown that mRNA transfer can generate "footprint-free" iPSCs, but these studies did not use a delivery vehicle and thus repetitive daily transfection was required because of mRNA degradation. Here, we report an mRNA delivery system employing graphene oxide (GO)-polyethylenimine (PEI) complexes for the efficient generation of "footprint-free" iPSCs. GO-PEI complexes were found to be very effective for loading mRNA of reprogramming transcription factors and protection from mRNA degradation by RNase. Dynamic suspension cultures of GO-PEI/RNA complexes-treated cells dramatically increased the reprogramming efficiency and successfully generated rat and human iPSCs from adult adipose tissue-derived fibroblasts without repetitive daily transfection. The iPSCs showed all the hallmarks of pluripotent stem cells including expression of pluripotency genes, epigenetic reprogramming, and differentiation into the three germ layers. These results demonstrate that mRNA delivery using GO-PEI-RNA complexes can efficiently generate "footprint-free" iPSCs, which may advance the translation of iPSC technology into the clinical settings.


Sorafenib inhibits liver regeneration in rats.
Sorafenib is a multikinase inhibitor with antiangiogenic and antiproliferative properties, approved for the treatment of hepatocellular carcinoma. The effect of Sorafenib on liver regeneration in healthy rats was investigated. Sixty Wistar rats received either Sorafenib (group S; 15 mg/kg) or placebo for 14 days prior to resection and until sacrifice. After a 70% partial hepatectomy, the rats were euthanized on post-operative days (POD) 2, 4 or 8. Hepatocyte proliferation was estimated by immunohistochemistry for Ki-67 antigen using stereological methods on sections prepared by systematic uniform random sampling. Seven animals (12%) died after surgery. Death rates were similar in treated rats and controls. At hepatectomy, the body weight was significantly lower in group S rats. The liver weight and regeneration rates were lower in group S rats on PODs 2, 4 and 8. Hepatocyte proliferation was significantly lower in group S animals on PODs 2 and 4. Alanine aminotransferase ALAT was significantly higher in the Sorafenib-treated group on PODs 2, 4 and 8. Alkaline phosphatase ALP and bilirubin levels were similar in the two groups, although bilirubin was elevated in group S rats on POD 8. In this rat model, Sorafenib did not increase post-hepatectomy mortality, but was associated with a significant impaired liver weight gain, regeneration rates and hepatocyte proliferation.


[Effects of intense pulse light on the mRNA expression of type I procollagen in human fibroblasts].
To investigate the effects of intense pulse light (IPL) on the mRNA expression of type I procollagen in human fibroblasts. Four healthy female subjects aged 19 - 41, underwent IPL on the back skin 3 times with an interval of 2 weeks. Before the IPL and after 1, 2, and 3 times of treatment samples of back skin were obtained to undergo real time quantitative PCR to detect the mRNA expression of type I procollagen. The mRNA expression level of type I procollagen after one time of IPL treatment was [(4.7 +/- 1.0) x 10(-4)], not significantly different from that before treatment [(2.1 +/- 0.7) x 10(-4), P = 0.100]. The mRNA expression levels of type I procollagen after treatment for 2 times and 3 -time were (7.9 +/- 1.7) x 10(-4) and (11.1 +/- 2.4) x 10(-4) respectively, both significantly higher than that before treatment (both P < 0.05). Promoting the mRNA transcription of type I procollagen in dermal fibroblasts, IPL therapy is effective in dermal remodeling and wrinkle removing.


Acquired hemolytic anemia after minor ABO incompatible renal transplantation.
Passenger lymphocytes in grafted kidney have been reported to result in autoimmune hemolytic anemia (AIHA) in minor ABO incompatible transplants. We followed up 15 cases of minor ABO incompatible transplants for one year. Two patients developed severe but self-limited auto-immune hemolytic anemia within two weeks of renal transplantation. Both were blood group A and had received a kidney from a group O donor. Preoperative cross-match and antibody screen was negative; however subsequent to the hemolytic episode, group-specific blood was incompatible and patients were transfused with group O, cross-match compatible blood. Serological tests showed positive direct anti-globulin test (DAT) and anti-A was eluted from both cases.


Water, land, fire, and forest: Multi-scale determinants of rainforests in the Australian monsoon tropics.
The small rainforest fragments found in savanna landscapes are powerful, yet often overlooked, model systems to understand the controls of these contrasting ecosystems. We analyzed the relative effect of climatic variables on rainforest density at a subcontinental level, and employed high-resolution, regional-level analyses to assess the importance of landscape settings and fire activity in determining rainforest density in a frequently burnt Australian savanna landscape. Estimates of rainforest density (ha/km2) across the Northern Territory and Western Australia, derived from preexisting maps, were used to calculate the correlations between rainforest density and climatic variables. A detailed map of the northern Kimberley (Western Australia) rainforests was generated and analyzed to determine the importance of geology and topography in controlling rainforests, and to contrast rainforest density on frequently burnt mainland and nearby islands. In the northwestern Australian, tropics rainforest density was positively correlated with rainfall and moisture index, and negatively correlated with potential evapotranspiration. At a regional scale, rainforests showed preference for complex topographic positions and more fertile geology. Compared with mainland areas, islands had significantly lower fire activity, with no differences between terrain types. They also displayed substantially higher rainforest density, even on level terrain where geomorphological processes do not concentrate nutrients or water. Our multi-scale approach corroborates previous studies that suggest moist climate, infrequent fires, and geology are important stabilizing factors that allow rainforest fragments to persist in savanna landscapes. These factors need to be incorporated in models to predict the future extent of savannas and rainforests under climate change.


Evaluation of the position of the posterior superior alveolar artery in relation to the maxillary sinus using the Cone-Beam computed tomography scans.
The aim of the present study was to evaluate the diameter, relationship and position of the posterior superior alveolar artery and its relationship with the alveolar ridge, the medial wall of the maxillary sinus, the prevalence of pathologic conditions and the maxillary sinus septa on CBCT images. A total of 200 CBCT images (400 maxillary sinuses) of patients over 20 years of age were evaluated. The distances between the lower border of the artery and the alveolar crest and between the artery and the medial wall of the sinus and the diameter of the artery were measured. The position of the artery, the presence of pathologic conditions and septa were recorded in the posterior region in: a) males edentulous in the posterior region; b) males having teeth in the posterior region; c) females edentulous in the posterior region; and d) females having teeth in the posterior region. The mean distance between the artery and the alveolar crest, irrespective of groupings, was 16.17±1.63 mm, with significant differences between the groups (P<0.05). The mean distance between the artery and the medial wall of the sinus was 11.65±1.21 mm, with no significant differences between the groups (P=0.796). The mean diameter of the canal was 1.37±0.44 mm, with no significant differences between the 4 groups (P=0.570). The position of the artery was intraosseous in 73.2%, beneath the sinus membrane in 21.7% and external to the lateral wall of the sinus in 4.9% of the cases. The overall prevalence rates of pathologic conditions and septa in the maxillary sinus were 45.7% and 26%, respectively. CBCT technique is useful for such evaluations and for possible variations in maxillary sinuses and presence of septa and pathologic entities in maxillary sinuses. Key words:Maxillary sinus, maxillary artery, Cone-Beam computed tomography.


Prolonged PD1 Expression on Neonatal Vδ2 Lymphocytes Dampens Proinflammatory Responses: Role of Epigenetic Regulation.
A successful pregnancy depends on the maintenance of tolerance at the fetal-maternal interface; strong inflammation in the placental bed is generally associated with adverse fetal outcomes. Among the mechanisms that foster tolerance and limit inflammation, the fetal immune system favors Th2 or regulatory responses over Th1 responses. The unintended consequence of this functional program is high susceptibility to infections. Human Vδ2 T cells mount innate-like responses to a broad range of microorganisms and are poised for Th1 responses before birth. In infants they likely play a key role in protection against pathogens by exerting early Th1 effector functions, improving function of other innate cells, and promoting Th1 polarization of adaptive responses. However, their propensity to release Th1 mediators may require careful regulation during fetal life to avoid exaggerated proinflammatory responses. We investigated molecules with the potential to act as a rheostat for fetal Vδ2 cells. Programmed death 1 (PD1) is a negative regulator of T cell responses and a determinant of tolerance, particularly at the fetal-maternal interface. Neonatal Vδ2 cells upregulate PD1 shortly after activation and, unlike their adult counterparts, express this molecule for at least 28 d. Engagement of PD1 by one of its ligands, PDL1, effectively dampens TCR-mediated responses (TNF-α production and degranulation) by neonatal Vδ2 cells and may thus help maintain their activity within safe limits. PD1 expression by neonatal Vδ2 cells is inversely associated with promoter DNA methylation. Prolonged PD1 expression may be part of a functional program to control Vδ2 cell inflammatory responses during fetal life.


Seroconversion and fever are dose-dependent in a nonhuman primate model of inhalational COVID-19.
While evidence exists supporting the potential for aerosol transmission of SARS-CoV-2, the infectious dose by inhalation remains unknown. In the present study, the probability of infection following inhalation of SARS-CoV-2 was dose-dependent in a nonhuman primate model of inhalational COVID-19. The median infectious dose, assessed by seroconversion, was 52 TCID50 (95% CI: 23-363 TCID50), and was significantly lower than the median dose for fever (256 TCID50, 95% CI: 102-603 TCID50), resulting in a group of animals that developed an immune response post-exposure but did not develop fever or other clinical signs of infection. In a subset of these animals, virus was detected in nasopharyngeal and/or oropharyngeal swabs, suggesting that infected animals without signs of disease are able to shed virus and may be infectious, which is consistent with reports of asymptomatic spread in human cases of COVID-19. These results suggest that differences in exposure dose may be a factor influencing disease presentation in humans, and reinforce the importance of public health measures that limit exposure dose, such as social distancing, masking, and increased ventilation. The dose-response data provided by this study are important to inform disease transmission and hazard modeling, and, ultimately, mitigation strategies. Additionally, these data will be useful to inform dose selection in future studies examining the efficacy of therapeutics and vaccines against inhalational COVID-19, and as a baseline in healthy, young adult animals for assessment of the importance of other factors, such as age, comorbidities, and viral variant, on the infectious dose and disease presentation.


Mathematical models of amino acid panel for assisting diagnosis of children acute leukemia.
The altered concentrations of amino acids were found in the bone marrow or blood of leukemia patients. Metabolomics technology combining mathematical model of biomarkers could be used for assisting the diagnosis of pediatric acute leukemia (AL). The concentrations of 17 amino acids was measured by targeted liquid chromatograph-tandem mass spectrometry in periphery blood collected using dried blood spots. After evaluation, the mathematical models were further evaluated by prospective clinical validation cohort for AL diagnosis. The concentrations of 13 in 17 amino acids were statistically different between the periphery blood dried serum dots measured by targeted LC-MS/MS. The receiver operating characteristic analysis for the models of amino acid panel showed that the area under curve for AL diagnosis were 0.848, 0.834 and 0.856 by SVM, RF and XGBoost. The Kappa values in further prospectively evaluated clinical cohort were 0.697, 0.703 and 0.789 (p > 0.05) respectively, and the accuracies for the models were 84.86%, 85.20% and 89.46% respectively with further clinical validation. The established mathematical model is a faster, cheaper and more convenient way than conventional methods, and no significant difference on the effect of diagnosis comparing with conventional methods. The mathematical model can be clinically useful for assisting pediatric AL diagnosis.


Human immunodeficiency virus can infect CD4-negative human fibroblastoid cells.
Certain human immunodeficiency virus (HIV) strains were shown to infect human fibroblastoid cells. Replication was demonstrated only by coculturing normal peripheral blood mononuclear cells or human T-lymphotropic virus type 1-transformed T-cell lines with the infected human cells. This infection of human fibroblastoid cells did not involve the CD4 molecule and did not have the properties of endocytosis. Human sera could be distinguished by their ability to neutralize HIV infection of the fibroblastoid versus human T-cell lines. These observations demonstrate further that other mechanisms for viral entry, besides CD4 binding, must be considered for HIV. They also indicate the wide cellular host range and heterogeneity of HIV strains. The possibility that fibroblastoid cells serve as a reservoir for the AIDS virus and are involved in connective tissue disorders of infected individuals merits attention.


The effects of simvastatin on the incidence of heart failure in patients with coronary heart disease.
Although treatment of myocardial overload effectively reduces death from progression of heart failure, it is not known whether the retardation of progressive coronary artery disease obtained with lipid lowering treatment will prevent the onset and consequences of heart failure in patients without previous symptoms of heart failure. In the Scandinavian Simvastatin Survival Study, 4444 patients with coronary heart disease without evidence of heart failure were randomized to placebo (n = 2223) or simvastatin 20-40 mg (n = 2221) and followed for more than 5 years. Among the patients who received placebo, 228 (10.3%) were diagnosed with heart failure during follow-up evaluation compared with 184 (8.3%) of patients who received simvastatin (P < .015). Mortality was 31.9% in the placebo group and 25.5% in the simvastatin group among patients who developed heart failure. These compare with 9.2 and 6.6%, respectively, among non-heart failure patients. There were 45 hospitalizations because of acute heart failure in the placebo group and 23 in the simvastatin group (NS). Patients who developed heart failure were more likely to have suffered a recurrent myocardial infarction and have a history of diabetes, peripheral artery disease, and hypertension than patients who did not develop congestive heart failure. Long-term prevention with simvastatin reduces the occurrence of heart failure in a cohort of patients with coronary heart disease without previous evidence of congestive heart failure.


Effects of muscle injury severity on localized bioimpedance measurements.
Muscle injuries in the lower limb are common among professional football players. Classification is made according to severity and is diagnosed with radiological assessment as: grade I (minor strain or minor injury), grade II (partial rupture, moderate injury) and grade III (complete rupture, severe injury). Tetrapolar localized bioimpedance analysis (BIA) at 50 kHz made with a phase-sensitive analyzer was used to assess damage to the integrity of muscle structures and the fluid accumulation 24 h after injury in 21 injuries in the quadriceps, hamstring and calf, and was diagnosed with magnetic resonance imaging (MRI). The aim of this study was to identify the pattern of change in BIA variables as indicators of fluid [resistance (R)] and cell structure integrity [reactance (Xc) and phase angle (PA)] according to the severity of the MRI-defined injury. The % difference compared to the non-injured contralateral muscle also measured 24-h after injury of R, Xc and PA were respectively: grade I (n = 11; -10.4, -17.5 and -9.0%), grade II (n = 8; -18.4, -32.9 and -16.6%) and grade III (n = 2; -14.1, -52.9 and -43.1%), showing a greater significant decrease in Xc (p < 0.001). The greatest relative changes were in grade III injuries. However, decreases in R, that indicate fluid distribution, were not proportional to the severity of the injury. Disruption of the muscle structure, demonstrated by the localized determination of Xc, increased with the severity of muscle injury. The most significant changes 24 h after injury was the sizeable decrease in Xc that indicates a pattern of disrupted soft tissue structure, proportional to the severity of the injury.


Development of a modified diagnostic classification system for voice disorders with inter-rater reliability study.
Diversity in nomenclature and on-going dilemmas over the conceptual bases for the classification of voice disorders make it virtually impossible for the collation and accurate comparison of evidence-based data across different clinical settings. This has significant implications for treatment outcome studies. The first aim of this study was to develop a modified diagnostic classification system for voice disorders with clearly defined operational guidelines by which we might reliably distinguish voice disorders from one another. The second aim was to establish the face validity and reliability of the system as an effective diagnostic tool for the allocation of patients to different diagnostic groups for clinical and research purposes. After the Diagnostic Classification System for Voice Disorders (DCSVD) had been developed, it was used in an inter-rater reliability study for the independent assessment of 53 new consecutive patients referred to the Voice Analysis Clinics of three tertiary hospitals. There were three raters present for the assessment and diagnostic allocation of each patient. The high levels of inter-rater reliability suggest this may be a robust classification system that has good face validity and even at this early stage, strong construct validity.


Rubisco condensate formation by CcmM in β-carboxysome biogenesis.
Cells use compartmentalization of enzymes as a strategy to regulate metabolic pathways and increase their efficiency1. The α- and β-carboxysomes of cyanobacteria contain ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco)-a complex of eight large (RbcL) and eight small (RbcS) subunits-and carbonic anhydrase2-4. As HCO3- can diffuse through the proteinaceous carboxysome shell but CO2 cannot5, carbonic anhydrase generates high concentrations of CO2 for carbon fixation by Rubisco6. The shell also prevents access to reducing agents, generating an oxidizing environment7-9. The formation of β-carboxysomes involves the aggregation of Rubisco by the protein CcmM10, which exists in two forms: full-length CcmM (M58 in Synechococcus elongatus PCC7942), which contains a carbonic anhydrase-like domain8 followed by three Rubisco small subunit-like (SSUL) modules connected by flexible linkers; and M35, which lacks the carbonic anhydrase-like domain11. It has long been speculated that the SSUL modules interact with Rubisco by replacing RbcS2-4. Here we have reconstituted the Rubisco-CcmM complex and solved its structure. Contrary to expectation, the SSUL modules do not replace RbcS, but bind close to the equatorial region of Rubisco between RbcL dimers, linking Rubisco molecules and inducing phase separation into a liquid-like matrix. Disulfide bond formation in SSUL increases the network flexibility and is required for carboxysome function in vivo. Notably, the formation of the liquid-like condensate of Rubisco is mediated by dynamic interactions with the SSUL domains, rather than by low-complexity sequences, which typically mediate liquid-liquid phase separation in eukaryotes12,13. Indeed, within the pyrenoids of eukaryotic algae, the functional homologues of carboxysomes, Rubisco adopts a liquid-like state by interacting with the intrinsically disordered protein EPYC114. Understanding carboxysome biogenesis will be important for efforts to engineer CO2-concentrating mechanisms in plants15-19.


A wireless sensor enabled by wireless power.
Through harvesting energy by wireless charging and delivering data by wireless communication, this study proposes the concept of a wireless sensor enabled by wireless power (WPWS) and reports the fabrication of a prototype for functional tests. One WPWS node consists of wireless power module and sensor module with different chip-type sensors. Its main feature is the dual antenna structure. Following RFID system architecture, a power harvesting antenna was designed to gather power from a standard reader working in the 915 MHz band. Referring to the Modbus protocol, the other wireless communication antenna was integrated on a node to send sensor data in parallel. The dual antenna structure integrates both the advantages of an RFID system and a wireless sensor. Using a standard UHF RFID reader, WPWS can be enabled in a distributed area with a diameter up to 4 m. Working status is similar to that of a passive tag, except that a tag can only be queried statically, while the WPWS can send dynamic data from the sensors. The function is the same as a wireless sensor node. Different WPWSs equipped with temperature and humidity, optical and airflow velocity sensors are tested in this study. All sensors can send back detection data within 8 s. The accuracy is within 8% deviation compared with laboratory equipment. A wireless sensor network enabled by wireless power should be a totally wireless sensor network using WPWS. However, distributed WPWSs only can form a star topology, the simplest topology for constructing a sensor network. Because of shielding effects, it is difficult to apply other complex topologies. Despite this limitation, WPWS still can be used to extend sensor network applications in hazardous environments. Further research is needed to improve WPWS to realize a totally wireless sensor network.


Pro-convulsant effect of cefazolin sodium against pentylenetetrazol- or picrotoxin-induced convulsions in mice.
Cefazolin injection (3000 mg/kg, i.v.) in mice showed several behavioral excitations such as wild running, jumping, rolling, and finally undergoing severe convulsions followed by death. It's lower doses (500-2000 mg/kg, i.v.) were unable to produce any convulsions or behavioral excitations in mice. However, cefazolin (500 or 1000 mg/kg, i.v.) when administered before different doses of pentylenetetrazol (PTZ; 40 or 60 mg/kg, i.p.) or picrotoxin (PTX; 4 or 8 mg/kg, i.p.), it produced severe tonic-clonic convulsions in mice. The convulsions or behavioral excitations produced by 3000 mg/kg, i.v. cefazolin was also reversed by different doses of diazepam (0.5-2 mg/kg, i.p.) further proving the GABAergic modulatory effect of cefazolin. The results conclude the pro-convulsant action of cefazolin on PTZ- or PTX-induced convulsions, and further confirm the clinical reports.


Dual-carrier processing to convey temporal fine structure cues: Implications for cochlear implants.
Speech intelligibility in noise can be degraded by using vocoder processing to alter the temporal fine structure (TFS). Here it is argued that this degradation is not attributable to the loss of speech information potentially present in the TFS. Instead it is proposed that the degradation results from the loss of sound-source segregation information when two or more carriers (i.e., TFS) are substituted with only one as a consequence of vocoder processing. To demonstrate this segregation role, vocoder processing involving two carriers, one for the target and one for the background, was implemented. Because this approach does not preserve the speech TFS, it may be assumed that any improvement in intelligibility can only be a consequence of the preserved carrier duality and associated segregation cues. Three experiments were conducted using this "dual-carrier" approach. All experiments showed substantial sentence intelligibility in noise improvements compared to traditional single-carrier conditions. In several conditions, the improvement was so substantial that intelligibility approximated that for unprocessed speech in noise. A foreseeable and potentially promising implication for the dual-carrier approach involves implementation into cochlear implant speech processors, where it may provide the TFS cues necessary to segregate speech from noise.


Is circadian variation in theophylline trough serum concentrations determined by time of dosing?
Ten healthy volunteers were each given two separate courses of eight doses of theophylline 200 mg as a slow release preparation at 12 hourly intervals. In the first course (regimen 1) the dose was given at 11.00 h and 23.00 h and in the second (regimen 2) at 05.00 h and 17.00 h. Sixty hours after the start of each course, four consecutive trough serum theophylline concentrations were measured. On regimen 1 the trough concentration of theophylline was 6.2 +/- 2.2 mg/l (mean +/- s.d.) at 11.00 h compared with 5.4 +/- 1.9 mg/l at 23.00 h (P less than 0.01). This circadian variation was abolished during regimen 2 when the mean theophylline concentrations were identical at 5.2 mg/l. The higher trough concentrations after the evening dose on regimen 1 and the loss of circadian variation on regimen 2 may be due to the change in sleep period in relation to the time of dosing.


Pulmonary embolism with vertebral augmentation procedures.
With the prevalence of an aging American population on the rise, osteoporotic vertebral fractures are becoming a common occurrence, resulting in an increase in vertebral augmentation procedures and associated complications such as cement leakage, vertebral compressions, and pulmonary embolism. We describe a patient who presented with respiratory distress three years following kyphoplasty of the lumbar vertebra. Computed tomography (CT) angiogram of the chest confirmed the presence of polymethylmethacrylate (PMMA) cement in the lung fields and pulmonary vessels. We conducted a systematic review of the published literature identifying effective management strategies for the treatment of vertebroplasty-associated pulmonary embolism.


Diagnostic Challenge of Acute Hepatitis in Human Immunodeficiency Virus-Positive Patients: With or Without Tuberculosis.
Tuberculosis (TB) is the leading infectious disease that causes death worldwide, eclipsing HIV/AIDS. It may affect any organ, but the most common manifestation is related to the involvement of the lungs. Hepatic tuberculosis is often a manifestation of disseminated disease and less likely a localized disease. Our case illustrates an HIV-positive patient with disseminated tuberculosis that presented first as liver involvement. After the diagnosis was made through liver biopsy, pulmonary compromise ensued. We review the clinical presentation, diagnosis, and treatment options of disseminated and hepatic TB. Our case is a glimpse of the many faces TB can adopt, especially in HIV-positive patients.


TCW: transcriptome computational workbench.
The analysis of transcriptome data involves many steps and various programs, along with organization of large amounts of data and results. Without a methodical approach for storage, analysis and query, the resulting ad hoc analysis can lead to human error, loss of data and results, inefficient use of time, and lack of verifiability, repeatability, and extensibility. The Transcriptome Computational Workbench (TCW) provides Java graphical interfaces for methodical analysis for both single and comparative transcriptome data without the use of a reference genome (e.g. for non-model organisms). The singleTCW interface steps the user through importing transcript sequences (e.g. Illumina) or assembling long sequences (e.g. Sanger, 454, transcripts), annotating the sequences, and performing differential expression analysis using published statistical programs in R. The data, metadata, and results are stored in a MySQL database. The multiTCW interface builds a comparison database by importing sequence and annotation from one or more single TCW databases, executes the ESTscan program to translate the sequences into proteins, and then incorporates one or more clusterings, where the clustering options are to execute the orthoMCL program, compute transitive closure, or import clusters. Both singleTCW and multiTCW allow extensive query and display of the results, where singleTCW displays the alignment of annotation hits to transcript sequences, and multiTCW displays multiple transcript alignments with MUSCLE or pairwise alignments. The query programs can be executed on the desktop for fastest analysis, or from the web for sharing the results. It is now affordable to buy a multi-processor machine, and easy to install Java and MySQL. By simply downloading the TCW, the user can interactively analyze, query and view their data. The TCW allows in-depth data mining of the results, which can lead to a better understanding of the transcriptome. TCW is freely available from www.agcol.arizona.edu/software/tcw.


The John Insall Award: unicompartmental knee replacement: a minimum twenty-one-year followup, end-result study.
We report the results of a minimum 21-year followup of a consecutive series of 103 patients who had 136 Marmor cemented unicompartmental knee replacements done between 1975 and 1982. Patients were evaluated clinically and radiographically. At minimum 21-year followup 14 patients (19 knees) were alive, 87 patients (115 knees) had died, and only two patients (two knees) were lost to followup. The average age at surgery was 70.9 years. The average followup Hospital for Special Surgery knee score was 58. The average Knee Society final followup clinical and functional scores averaged 72 and 53 points, respectively. Nineteen knees (14%) were revised during the 21-year followup period: nine for progression of disease, eight for loosening, and two for pain, at an average of 10.6 years (range, 1-22 years). Of the 19 knees in the 14 patients who were still alive at final followup, seven (37%) were revised: two for tibial loosening, four for disease progression, and one for pain. Unicompartmental knee replacements in this relatively older age group of patients performed well at minimum 21-year followup. Although we are encouraged by these results, only 22% were done in patients who were younger than 65 years at the time of surgery and the results in this group were significantly less durable. Therapeutic study, Level IV-1 (case series). See the Guidelines for Authors for a complete description of levels of evidence.


Prediction of nitroxide hyperfine coupling constants in solution from combined nanosecond scale simulations and quantum computations.
We present a combined theoretical approach based on analyzing molecular dynamics trajectories (at the nanosecond scale) generated by use of classical polarizable force fields and on quantum calculations to compute averaged hyperfine coupling constants. That method is used to estimate the constant of a prototypical nitroxide: the dimethylnitroxide. The molecule is embedded during the simulations in a cubic box containing about 500 water molecules and the molecular dynamics is generated using periodic conditions. Once the trajectories are achieved, the nitroxide and its first hydration shell molecules are extracted, and the coupling constants are computed by considering the latter aggregates by means of quantum computations. However, all the water molecules of the bulk are also accounted for during those computations by means of the electrostatic potential fitted method. Our results exhibit that in order to predict accurate and reliable coupling constants, one needs to describe carefully the out-of-plane motion of the nitroxide nitrogen and to sample trajectories with a time interval of 400 fs at least to generate an uncorrelated large set of nitroxide structures. Compared to Car-Parrinello molecular dynamics techniques, our approach can be used readily to compute hyperfine coupling constants of large systems, such as nitroxides of great size interacting with macromolecules such as proteins or polymers.


Development of octreotide-conjugated polymeric prodrug of bufalin for targeted delivery to somatostatin receptor 2 overexpressing breast cancer in vitro and in vivo.
Development of polymeric prodrugs of small molecular anticancer drugs has become one of the most promising strategies to overcome the intrinsic shortcomings of small molecular anticancer drugs and improve their anticancer performance. In the current work, we fabricated a novel octreotide (Oct)-modified esterase-sensitive tumor-targeting polymeric prodrug of bufalin (BUF) and explored its anticancer performance against somatostatin receptor 2 overexpressing breast cancer. The obtained tumor-targeting polymeric prodrug of BUF, P(oligo[ethylene glycol] monomethyl ether methacrylate [OEGMA]-co-BUF-co-Oct), showed a nanosize dimension and controlled drug release features in the presence of esterase. It was demonstrated by in vitro experiment that P(OEGMA-co-BUF-co-Oct) showed enhanced cytotoxicity, cellular uptake, and apoptosis in comparison with those of free BUF. In vivo experiment further revealed the improved accumulation of drugs in tumor tissues and enhanced anticancer performance of P(OEGMA-co-BUF-co-Oct). Taken together, this study indicated that polymeric prodrug of BUF holds promising potential toward the treatment of somatostatin receptor 2 overexpressing breast cancer.


Characterization of protein interactions with positive and negative elements regulating the apoVLDLII gene.
Synthesis of avian apo very-low-density lipoprotein (apoVLDL)II is estrogen dependent and liver specific. Competence to express the apoVLDLII gene is not acquired until days 7-9 of embryogenesis and thus lags 5-6 days behind appearance of the liver primordial bud. It is not known whether the delayed ability to activate the gene is attributable to hepatic estrogen receptor profiles, or a requirement for other transcription factors not expressed at earlier stages of embryogenesis. The latter possibility is supported by developmental alterations in nuclease hypersensitivity flanking the gene that occur independently of estrogen administration. We have examined the influence of these hypersensitive regions on expression from the apoVLDLII promoter and have characterized novel protein-DNA interactions at two of them. One is located in a copy of the CR1 family of middle repetitive elements approximately 3.0 kb upstream from the start of the gene. We demonstrate by DNase I footprinting that the site contains an element which matches a predicted consensus silencer sequence. The other site contains no previously identified binding motifs. It is located between nucleotides -228 and -245 and is adjacent to an imperfect estrogen response element (ERE) that we demonstrate acts additively with a canonical ERE 30 nucleotides downstream. We have identified ubiquitous and liver-specific factors that display overlapping DNA contacts with the site. Mutation of G residues contacted by these proteins decreases hormone-inducible expression from the promoter 5- to 8-fold. Hepatic levels of the liver-enriched factor interacting with this site increase abruptly between days 7 and 9 of embryogenesis, suggesting that it may be an important determinant of the ability to express the apoVLDLII and possibly other liver-specific genes.


Phyto-management of Cr-contaminated soils by sunflower hybrids: physiological and biochemical response and metal extractability under Cr stress.
Chromium (Cr) is a biologically non-essential, carcinogenic and toxic heavy metal. The cultivation of Cr-tolerant genotypes seems the most favorable and environment friendly strategy for rehabilitation and remediation of Cr-contaminated soils. To prove this hypothesis and identify the Cr tolerance, the present study was performed to assess the physiological and biochemical response of sunflower genotypes to Cr stress. The seeds of six sunflower hybrids, namely FH-425, FH-600, FH-612, FH-614, FH-619, and FH-620, were grown in spiked soil for 12 weeks under increasing concentrations of Cr (0, 5, 10, and 20 mg kg-1). A seed germination test was also run under different concentrations of Cr (0, 5, 10, 200 mM) in petri dishes. Plants were harvested after 12 weeks of germination. Different plant attributes such as growth; biomass; photosynthesis; gas exchange; activity of antioxidant enzymes, i.e., superoxide dismutase (SOD), guaiacol peroxidase (POD), ascorbate (APX), and catalases (CAT); reactive oxygen species (ROS); lipid peroxidation; electrolyte leakage; and Cr concentration as well as accumulations in all plant parts were studied for the selection of the most Cr-tolerant genotype. Increasing concentration of Cr in soil triggered the reduction of all plant parameters in sunflower. Cr stress increased electrolyte leakage and production of reactive oxygen species which stimulated the activities of antioxidant enzymes and gas exchange attributes of sunflower. Chromium accumulation in the root and shoot increased gradually with increasing Cr treatments and caused reduction in overall plant growth. The accumulation of Cr was recorded in the order of FH-614 > FH-620 > FH-600 > FH-619 > FH-612 > FH-425. The differential uptake and accumulation of Cr by sunflower hybrids may be useful in selection and breeding for Cr-tolerant genotypes.


Prion-like activity of Cu/Zn superoxide dismutase: implications for amyotrophic lateral sclerosis.
Neurodegenerative diseases belong to a larger group of protein misfolding disorders, known as proteinopathies. There is increasing experimental evidence implicating prion-like mechanisms in many common neurodegenerative disorders, including Alzheimer disease, Parkinson disease, the tauopathies, and amyotrophic lateral sclerosis (ALS), all of which feature the aberrant misfolding and aggregation of specific proteins. The prion paradigm provides a mechanism by which a mutant or wild-type protein can dominate pathogenesis through the initiation of self-propagating protein misfolding. ALS, a lethal disease characterized by progressive degeneration of motor neurons is understood as a classical proteinopathy; the disease is typified by the formation of inclusions consisting of aggregated protein within and around motor neurons that can contribute to neurotoxicity. It is well established that misfolded/oxidized SOD1 protein is highly toxic to motor neurons and plays a prominent role in the pathology of ALS. Recent work has identified propagated protein misfolding properties in both mutant and wild-type SOD1, which may provide the molecular basis for the clinically observed contiguous spread of the disease through the neuroaxis. In this review we examine the current state of knowledge regarding the prion-like properties of SOD1 and comment on its proposed mechanisms of intercellular transmission.


OAZ1 knockdown enhances viability and inhibits ER and LHR transcriptions of granulosa cells in geese.
An increasing number of studies suggest that ornithine decarboxylase antizyme 1 (OAZ1), which is regarded as a tumor suppressor gene, regulates follicular development, ovulation, and steroidogenesis. The granulosa cells in the ovary play a critical role in these ovarian functions. However, the action of OAZ1 mediating physiological functions of granulosa cells is obscure. OAZ1 knockdown in granulosa cells of geese was carried out in the current study. The effect of OAZ1 knockdown on polyamine metabolism, cell proliferation, apoptosis, and hormone receptor transcription of primary granulosa cells in geese was measured. The viability of granulosa cells transfected with the shRNA OAZ1 at 48 h was significantly higher than the control (p<0.05). The level of putrescine and spermidine in granulosa cells down-regulating OAZ1 was 7.04- and 2.11- fold higher compared with the control, respectively (p<0.05). The CCND1, SMAD1, and BCL-2 mRNA expression levels in granulosa cells down-regulating OAZ1 were each significantly higher than the control, respectively (p<0.05), whereas the PCNA and CASPASE 3 expression levels were significantly lower than the control (p<0.05). The estradiol concentration, ER and LHR mRNA expression levels were significantly lower in granulosa cells down-regulating OAZ1 compared with the control (p<0.05). Taken together, our results indicated that OAZ1 knockdown elevated the putrescine and spermidine contents and enhanced granulosa cell viability and inhibited ER and LHR transcriptions of granulosa cells in geese.


Antitumor agents. 7. Synthesis and antitumor activity of novel hexacyclic camptothecin analogues.
Eleven novel hexacyclic and three 7,9-disubstituted pentacyclic derivatives of camptothecin were synthesized and evaluated for in vitro cytotoxic activity against P388, HOC-21, and QG-56 and in vivo antileukemic activity against P388 in mice. Hexacyclic compounds which have an additional 5-, 6-, or 7-membered ring cyclized at positions 7 and 9 of camptothecin were prepared by intramolecular cyclization of pentacyclic camptothecin derivatives or Friedländer condensation of the appropriate bicyclic amino ketone and tricyclic ketone. All of the hexacyclic compounds exhibited compatible or superior activity of 7-ethyl-10-hydroxycamptothecin (SN-38) in in vitro assays without regard to the size or type of the additional ring, and three of six compounds showed more than 300% T/C on in vivo assay. These results suggest that the potency of the hexacyclic ring system is higher than that of the original pentacyclic ring system of camptothecin and that the conformational rigidity of substituents at positions 7 and 9 is favorable for antitumor activity.


Bmi-1 is useful as a novel molecular marker for predicting progression of myelodysplastic syndrome and patient prognosis.
The International Prognostic Scoring System (IPSS) has been widely used to predict the prognosis of patients with myelodysplastic syndrome (MDS). However, IPSS does not always provide a sufficiently precise evaluation of patients to allow the appropriate choice of clinical interventions. Here, we analyzed the expression of Bmi-1, which is required to regulate the self-renewal in CD34+ cells from 51 patients with cases of MDS and acute myeloid leukemia preceded by MDS (MDS-AML). Higher positivity rate of Bmi-1 was preferentially seen in refractory anemia with excess blasts (RAEB), RAEB in transformation (RAEB-T), and MDS-AML compared with refractory anemia (RA) and RA with ringed sideroblasts (RARS). IPSS score was positively correlated with the percentage of Bmi-1 expression. Patients with RA and RARS with a higher percentage of Bmi-1+ cells showed disease progression to RAEB. Here, we propose Bmi-1 as a novel molecular marker to predict the progression and prognosis of MDS.


Joint cracking and popping: understanding noises that accompany articular release.
Articular release is a physiologic event that may or may not be audible. It is seen in patients with healthy joints as well as those with somatic dysfunction. After an articular release, there is a difference in joint spacing-with the release increasing the distance between articular surfaces. Not all noise that emanates from a joint signifies an articular release. A hypothesis about the noise that frequently accompanies this release is offered and includes anatomic, physiologic, and functional models of articular release. Repeated performance of articular release may decrease the occurrence of arthritis. Potential problems from repeated articular release (eg, hypermobility) are also examined.


Microfluidic Droplet-Storage Array.
A microfluidic droplet-storage array that is capable of the continuous operation of droplet formation, storing, repositioning, retrieving, injecting and restoring is demonstrated. The microfluidic chip comprised four valve-assisted droplet generators and a 3 × 16 droplet-storage array. The integrated pneumatically actuated microvalves enable the precise control of aqueous phase dispensing, as well as carrier fluid flow path and direction for flexible manipulating water-in-oil droplets in the chip. The size of droplets formed by the valve-assisted droplet generators was validated under various operating conditions such as pressures for introducing solutions and dispensing time. In addition, flexible droplet addressing in the storage array was demonstrated by storing droplets with various numbers and compositions in different storage units as well as rearranging their stored positions. Moreover, serial injections of new droplets into a retrieved droplet from a storage unit was performed to show the potential of the platform in sequential dosing on incubated droplet-based reactors at the desired timeline. The droplet-storage array with great freedom and flexibility in droplet handling could be applied for performing complex chemical and biologic reactions, especially in which incubation and dosing steps are necessary.


A review of data on the U.S. health sector spring 2006.
This report presents information on the state of the U.S. health system in the spring of 2006. It includes data on the uninsured and underinsured and their access to health care, on socioeconomic inequality in health care, and on the rising costs of the U.S. health system. It also presents information on the role of corporate money in health care, focusing on the pharmaceutical industry, Medicare HMOs, and corporate-government conflicts of interest. The author includes a survey of recent public opinion polls on health care and health system reform and an update on the U.S. national health insurance legislation. The article ends by reviewing recent data on international health systems and international system comparisons.


Role of epidermal growth factor expression in early mouse embryo lung branching morphogenesis in culture: antisense oligodeoxynucleotide inhibitory strategy.
Epidermal growth factor (EGF) expression and branching morphogenesis were inhibited using a 5' 15-mer antisense oligodeoxynucleotide (ODN) directed against EGF precursor mRNA in embryonic mouse lung in culture under chemically defined, serumless conditions. Antisense EGF ODN resulted in > 90% inhibition of EGF immunoreactive peptide synthesis, 75% reduction in branching morphogenesis, 73% decrease in DNA content, 64% decrease in RNA content, 73% decrease in protein synthesis, and 65% decrease in [3H]thymidine incorporation into DNA compared to Embryonic Day 11 controls in culture for 4 days. Sense ODN results were similar to control. Supplementing antisense ODN with EGF partially reversed antisense effects. The results further support a role for EGF in pulmonary organogenesis.


The association of oxidative stress and disease activity in seborrheic dermatitis.
The pathogenesis of seborrheic dermatitis (SD) has not been clearly identified, and many factors are thought to play a role in its development. Recently, new studies have focused on increased oxidative stress (OS) in T cell-mediated skin diseases like psoriasis, contact dermatitis, and atopic dermatitis. However, there is no study investigating the status of OS in SD. In this study, we aimed to determine the status of OS in SD and the correlation of disease severity with OS. Fifty-four patients who were clinically and/or histopathologically diagnosed with SD were included in the study. Fifty-four healthy volunteers constituted the control group. Disease severity in patients with SD was scored according to the Seborrheic Dermatitis Area and Severity Index (SDASI). Serum total antioxidant status (TAS) and total oxidative status (TOS) were measured, and the oxidative stress index (OSI) was calculated in all patients and control subjects. The mean TAS values were significantly lower in the patient group than in the control group (p = 0.024). However, patients had significantly higher TOS and OSI values than the controls (p < 0.05). There was no correlation between SDASI and TAS, TOS, and OSI values. In this study, the association of oxidative stress and disease activity has first investigated in seborrheic dermatitis. It was found that OS was significantly higher in SD patients than in healthy subjects. In conclusion, our findings point to the possible role of the OS for the etiopathogenesis of SD.


Traffic-based adaptive bandwidth adjustment for flexible OTN connectivity in optical networks.
In conventional optical transport networks, the service form is the fixed bandwidth connectivity, which is not flexible for carrying bursting traffic. To support the time-varying traffic in an efficient way, researchers are studying the optical service units for building the more flexible optical transport network (OTN) connectivity, which is capable of dynamic hitless bandwidth adjustment. To better utilize the benefits of flexible connectivity, network operators need efficient algorithms to adjust the flexible connectivity bandwidth, especially in the network with a massive number of connections. In this paper, based on max-min fair bandwidth allocation criteria, we propose two traffic-based adaptive bandwidth adjustment algorithms to make bandwidth adjustment decisions, with the aim to improve bandwidth effectiveness. Simulation results show that the proposed algorithms can improve bandwidth utilization by up to 16%. Additionally, under high load conditions, it can reduce the loss in traffic bitrate by a maximum of 10%.


[The expression and correlation of inducible nitric oxide synthase, Bax and Bcl-2 in nasal polyps].
To detect the expressions of inducible nitric oxide synthase (iNOS), apoptosis-related gene Bax, Bcl-2 in nasal polyps,and discuss the their relationship. The apoptosis of 30 cases of nasal polyps was detected by TUNEL assay. The expressions of iNOS and Bax, Bcl-2 was detected by immunohistochemical SABC method. The expressions of iNOS and Bax, Bcl-2 was measured by western blot. 1) The weakly positive stained apoptotic cells were detected at the surface epithelial cells and glandular epithelial cells of nasal polyps by TUNEL assay. 2) Immunohistochemical method revealed that positive stainings of iNOS, Bax, Bcl-2 located in the cytoplasm of epithelial cells, glandular epithelial cells, endothelial cells and infiltrating inflammatory cells in nasal polyps. The expression of Bax was weak, while the expressions of iNOS and Bcl-2 were strong. 3) iNOS, Bcl-2 and Bax was detected by western blot. The expressions of these proteins were significantly different (P<0.01). The expression of iNOS and Bcl-2 had a positive correlation (r=0.851, P<0.01), while the expression of iNOS and Bax had a negative correlation (r=-0.714, P<0.01). The pro-apoptotic and anti-apoptotic proteins are co-existed in the nasal polyps, iNOS may play an important role in the pathogens of nasal polyps through inhibition of apoptosis.


Single-pulse photoacoustic technique for measuring IR multiphoton absorption by polyatomic molecules.
The single-pulse photoacoustic technique was used to measure cross sections for IR multiphoton absorption. A discussion of this technique and the applicable pressure range is given. The source of the signals from the microphone was identified. The first peak is proportional to the energy absorbed by the gas in the focal region, while the subsequently arriving signals correspond to a mixture of the signal from the entrance and exit volumes and the ringings of the microphone and the gas cell.


Pitx2c attenuation results in cardiac defects and abnormalities of intestinal orientation in developing Xenopus laevis.
The experimental manipulation of early embryologic events, resulting in the misexpression of the homeobox transcription factor pitx2, is associated with subsequent defects of laterality in a number of vertebrate systems. To clarify the role of one pitx2 isoform, pitx2c, in determining the left-right axis of amphibian embryos, we examined the heart and gut morphology of Xenopus laevis embryos after attenuating pitx2c mRNA levels using chemically modified antisense oligonucleotides. We demonstrate that the partial depletion of pitx2c mRNA in these embryos results in alteration of both cardiac morphology and intestinal coiling. The most common cardiac abnormality seen was a failure of rightward migration of the outflow tract, while the most common intestinal laterality phenotype seen was a full reversal in the direction of coiling, each present in 23% of embryos injected with the pitx2c antisense oligonucleotide. An abnormality in either the heart or gut further predisposed to a malformation in the other. In addition, a number of other cardiac anomalies were observed after pitx2c mRNA attenuation, including abnormalities of atrial septation, extracellular matrix restriction, relative atrial-ventricular chamber positioning, and restriction of ventricular development. Many of these findings correlate with cardiac defects previously reported in pitx2 null and hypomorphic mice, but can now be assigned specifically to attenuation of the pitx2c isoform in Xenopus.


Robotic Assist-As-Needed as an Alternative to Therapist-Assisted Gait Rehabilitation.
Body Weight Supported Treadmill Training (BWSTT) with therapists' assistance is often used for gait rehabilitation post-stroke. However, this training method is labor-intensive, requiring at least one or as many as three therapists at once for manual assistance. Previously, we demonstrated that providing movement guidance using a performance-based robot-aided gait training (RAGT) that applies a compliant, assist-as-needed force-field improves gait pattern and functional walking ability in people post-stroke. In the current study, we compared the effects of assist-as-needed RAGT combined with functional electrical stimulation and visual feedback with BWSTT to determine if RAGT could serve as an alternative for locomotor training. Twelve stroke survivors were randomly assigned to one of the two groups, either receiving BWSTT with manual assistance or RAGT with functional electrical stimulation and visual feedback. All subjects received fifteen 40-minutes training sessions. Clinical measures, kinematic data, and EMG data were collected before and immediately after the training for fifteen sessions. Subjects receiving RAGT demonstrated significant improvements in their self-selected over-ground walking speed, Functional Gait Assessment, Timed Up and Go scores, swing-phase peak knee flexion angle, and muscle coordination pattern. Subjects receiving BWSTT demonstrated significant improvements in the Six-minute walk test. However, there was an overall trend toward improvement in most measures with both interventions, thus there were no significant between-group differences in the improvements following training. The current findings suggest that RAGT worked at least as well as BWSTT and thus may be used as an alternative rehabilitation method to improve gait pattern post-stroke as it requires less physical effort from the therapists compared to BWSTT.


[Characterization of the Russian beef cattle breed gene pools using inter simple sequence repeat DNA analysis (ISSR analysis)].
The gene pools of beef cattle breeds bred in Russia were characterized on the basis of inter simple sequence repeat DNA analysis (ISSR analysis). Samples of Aberdeen Angus, Kalmyk, and Kazakh Whitehead breeds from Russia, as well as of Hereford breed, hybrids of Kazakh Whitehead and Hereford breeds, and Kazakh Whitehead breed from the Republic of Kazakhstan, were examined. In the examined breeds, 27 AG-ISSR fragments were identified, 25 of which were polymorphic. The examined breeds were different both in the fragment profiles (the presence/absence of individual ISSR fragments) and in their frequencies. It was demonstrated that the hybrid animals lacked some ISSR fragments that were present with high frequencies in parental forms, suggesting considerable genome rearrangement in the hybrid animals (at the regions of microsatellite localization) in crossings of the individuals from different breeds. The level of genetic diversity in Russian beef breeds was consistent with the values typical of farmed populations (breeds). The genetic diversity parameters assessed by applying Nei's gene diversity index and the Shannon index varied from 0.0218 to 0.0605 and from 0.0225 to 0.0819, respectively. The highest Shannon index value was detected in the Kalmyk breed (0.0837) and Kazakh Whitehead breed from Russia (0.0819), and the highest level of Nei's gene diversity index was found in the Kalmyk breed (0.0562) and in both populations of the Kazakh Whitehead breed (0.0509 and 0.0605). The high level of genetic similarity (according to Nei) was revealed between Russian beef cattle breeds and Hereford cattle: 0.839 (for the Kazakh Whitehead breed from Russia) and 0.769 (for the Kalmyk breed).


Differences in the effects of chenodeoxycholic and ursodeoxycholic acid on biliary lipid secretion and bile acid synthesis in patients with gallstones.
Measurements of biliary lipid secretion rates were performed in 10 patients with radiolucent gallstones before and after 4 wk of administration of chenodeoxycholic acid and ursodeoxycholic acid (1 g/day) in a randomized crossover study. The results of both bile acid feeding periods were similar in many respects: expansion of the bile acid pool, increase in bile acid and phospholipid secretion, reduction in cholesterol output, and decrease in percent saturation of hepatic bile, which was more pronounced with ursodeoxycholic than chenodeoxycholic acid therapy. Despite these similarities, the mechanisms by which these two litholytic bile acids induced these changes were quite different. Ursodeoxycholic acid, in contrast to chenodeoxycholic acid, only partially suppresses bile acid synthesis. During chenodeoxycholic acid feeding, the ratio of phospholipids to bile acids increased from 0.264 to 0.307 (p less than 0.05), indicating an increased coupling of phospholipids by chenodeoxycholic acid, whereas ursodeoxycholic acid did not alter this ratio. The molar ratio of cholesterol to bile acid during the chenodeoxycholic- and ursodeoxycholic-acid periods decreased significantly from 0.073 to 0.058 and 0.041, respectively. However, this ratio during the ursodeoxycholic-acid period was unchanged when the amount of ursodeoxycholic acid was subtracted from total bile acid (0.069), indicating that UDCA has little, if any, effect on the mobilization of hepatic cholesterol into bile.


Morphology of the cochlear nucleus of the normal and reeler mutant mouse.
The morphology of the cochlear nuclei of normal and reeler mutant mice were studied in Nissl-stained sections. The cochlear nucleus in both mice is divisible into three parts: the anteroventral, posteroventral, and dorsal nuclei. Nine cell types can be recognized in the normal mouse. In the anteroventral nucleus spherical cells occupy the rostral pole. Globular cells are located caudally and extend to the interstitial region of the anteroventral nucleus. In the posteroventral nucleus multipolar cells are located rostrally and dark-staining cells occupy the caudal pole. Multipolar cells are also present in the anteroventral nucleus and in the deep region and molecular layer of the dorsal cochlear nucleus. The dorsal and lateral aspects of the ventral nuclei are covered by a granule cell layer. The dorsal nucleus consists of superficial molecular and pyramidal layers and a deep region. The deep region contains small and giant cells as well as multipolar cells. The pyramidal layer is made up of pyramidal cells, horizontal cells, and granule cells. Small cells are also present in the molecular layer and throughout the ventral nuclei. The dorsal cochlear nucleus of the reeler mutant mouse is disorganized and the molecular layer is reduced in thickness. The organization of the pyramidal layer is disrupted with granule cells superficial to pyramidal and horizontal cells. Cells which appear to be homologous to pyramidal cells are also present in the deep region of the dorsal nucleus. The total number of granule cells is reduced by an average of 42% over the whole nucleus and the reduction in granule cells is greatest in the granule cell cap covering the dorsal and lateral surface of the ventral cochlear nuclei. The cytoarchitecture of the ventral cochlear nucleus appears normal.


Changing trend in bacterial etiology and antibiotic resistance in sepsis of intramural neonates at a tertiary care hospital.
Septicemia is an important cause of neonatal morbidity and mortality. However, organized data on causative organisms and their resistant pattern are scanty from developing countries. The changing trend in causative organisms and their antibiotic resistance is yet to be documented in India. The present study examines the trends in bacterial profile and antibiotic resistance of the organisms causing sepsis in hospitalized neonates. A retrospective laboratory-based analysis of blood cultures obtained from Neonatal Intensive Care Unit of a tertiary care hospital in New Delhi was done for the period of 1999-2014, divided into five phases. A total of 4700 isolates were considered. Over time, Gram-negative organisms have replaced Gram-positives as frequent isolates. Initially, there was predominance of Klebsiella pneumoniae, then of Staphylococcus aureus which recently has been changed with coagulase negative-Staphylococcus and Acinetobacter. Growing resistance against the first and second line of drugs has been noted, including methicillin-resistant S. aureus and vancomycin-resistant Enterococcus. The etiological profile of neonatal sepsis has changed tremendously in the past 15 years. High resistance against common drugs necessitates continued surveillance and review of empirical antibiotic policy for neonatal sepsis. These steps are important to effectively curtail the surge of further antibiotic resistance.


[The prevent miscarriage effects of Yun Kang oral liquid on threatened abortion rat induced by kidney deficiency and luteum inhibition].
Yun Kang oral liquid is a listed proprietary Chinese Medicine. To further evaluate its efficacy, this experiment established a kidney deficiency and luteum inhibition threatened abortion rat model to observe the effects of Yun Kang oral liquid. Sixty pregnant rats were randomly divided into normal control group (NC), model group (MG), dydrogesterone group (DT, 3.02 mg/kg), and Yun Kang oral liquid low-dose group (YK-L, 4 ml/kg), medium dose group (YK-M, 6 ml/kg), high dose group (YK-H, 9 ml/kg), 10 in each group. On the first day of pregnancy, each administration group was treated with the test drug at the prescribed dose every morning, and the NC group and the MG group were given an equal volume of purified water for 10 days; the rats were intragastrically administrated every afternoon, except for the NC group. In addition, the other groups were intragastrically administered with hydroxyurea at a dose of 450 mg/kg for 9 days, and mifepristone was administered at a dose of 4.0 mg/kg on the 10th day. On the 9th day of pregnancy, behavioral signs such as back temperature, grasping force, pain threshold, and autonomic activity were measured in each group. On the 11th day of pregnancy, blood was collected from the abdominal aorta in each group to determine serum levels of estradiol (E2) , progesterone (P) and thromboxane B2 (TXB2) . Ovary and fetal uterus were removed, the number and diameter of embryos were observed, and the ovary and uterus indexes were calculated. Compared with the NC group, the back temperature, grip, pain threshold, number of spontaneous activities, number of embryos, embryo diameter, uterus index and serum E2, P, TXB2 levels in the MG group were decreased significantly (P<0.05, 0.01). Compared with the MG group, the back temperature, grasping force, number of embryos, embryo diameter and serum E2 and P levels were increased significantly in each dose group (P<0.05, 0.01); the pain threshold, autonomic activity, and uterus index of YK-M and YK-H group were increased significantly (P<0.05); serum level of TXB2 in YK-H group were increased significantly (P<0.05). Yun Kang oral liquid has a clear kidney-filling effect on rats with threatened abortion caused by kidney deficiency-luteal suppression. The mechanism may be related to raising serum E2, P, TXB2 levels, improving kidney deficiency and improving embryo quality.


Dobutamine induced eosinophilic myocarditis and right heart failure requiring emergent biventricular assist device implantation.
Dobutamine is a rare and unrecognized cause of eosinophilic myocarditis (EM). It is even more unique for it to cause significant clinical deterioration. Patients with end stage cardiomyopathy have very little cardiac reserve and dobutamine induced myocarditis may cause enough harm to require urgent mechanical circulatory support. We describe a man with dilated cardiomyopathy that developed accelerated clinical deterioration from dobutamine induced EM. He had a predominance of right heart failure, requiring an urgent biventricular assist device and acutely responded to withdrawal of dobutamine.


The State of Essential Newborn Care by Delivery Location in Bangladesh.
Introduction Essential newborn care (ENC) around the time of birth is critical in improving neonatal survival. There is currently a gap in our knowledge of the use of ENC by place of delivery in Bangladesh. This study assesses the provision of ENC and examines the odds of newborns receiving ENC by different levels of delivery care in Bangladesh. Methods Descriptive statistics and logistic regressions were performed on ENC practices from the 2011 Bangladesh Demographic and Health Survey dataset. ENC practices included nonapplication of substances to the cord; application of antiseptic to the cord; drying newborn within 5 min; wrapping newborn within 5 min; delaying first bath until the first 72 h; and breastfeeding within 1 h. Key predictors included home delivery with a lay attendant, delivery with primary healthcare services and delivery with higher-level healthcare services. Results Coverage of ENC practices was low. Women who delivered with primary and higher-level healthcare services generally reported greater odds of their newborns receiving recommended ENC than women who had home delivery with a lay attendant, the referent category. However, the odds of delayed first bath until 72 h and breastfeeding within 1 h were not statistically different for newborns who were delivered with primary healthcare services. Discussion These findings have significant public health implications as primary healthcare facilities are the first point of entry into the healthcare system. Provision of ENC, particularly delayed first bath until 72 h and breastfeeding within 1 h, should be encouraged for all healthy mother-newborn pairs in Bangladesh.


Hierarchy of orientational phases and axial anisotropies in the gauge theoretical description of generalized nematic liquid crystals.
The paradigm of spontaneous symmetry breaking encompasses the breaking of the rotational symmetries O(3) of isotropic space to a discrete subgroup, i.e., a three-dimensional point group. The subgroups form a rich hierarchy and allow for many different phases of matter with orientational order. Such spontaneous symmetry breaking occurs in nematic liquid crystals, and a highlight of such anisotropic liquids is the uniaxial and biaxial nematics. Generalizing the familiar uniaxial and biaxial nematics to phases characterized by an arbitrary point-group symmetry, referred to as generalized nematics, leads to a large hierarchy of phases and possible orientational phase transitions. We discuss how a particular class of nematic phase transitions related to axial point groups can be efficiently captured within a recently proposed gauge theoretical formulation of generalized nematics [K. Liu, J. Nissinen, R.-J. Slager, K. Wu, and J. Zaanen, Phys. Rev. X 6, 041025 (2016)2160-330810.1103/PhysRevX.6.041025]. These transitions can be introduced in the model by considering anisotropic couplings that do not break any additional symmetries. By and large this generalizes the well-known uniaxial-biaxial nematic phase transition to any arbitrary axial point group in three dimensions. We find in particular that the generalized axial transitions are distinguished by two types of phase diagrams with intermediate vestigial orientational phases and that the window of the vestigial phase is intimately related to the amount of symmetry of the defining point group due to inherently growing fluctuations of the order parameter. This might explain the stability of the observed uniaxial-biaxial phases as compared to the yet to be observed other possible forms of generalized nematic order with higher point-group symmetries.


Charge density study of hydrogen [(2,4-diaminopyrimidin-1-io)methyl]phosphonate monohydrate.
The crystal structure and charge density of hydrogen (2,4-diaminopyrimidin-1-io)methyl]phosphonate monohydrate, C(5)H(9)N(4)O(3)P x H(2)O, have been determined by means of single-crystal X-ray diffraction. Diffraction data were collected at 105 K with Mo K alpha radiation to a resolution of sin theta/lambda = 1.08 A(-1). A four-circle diffractometer equipped with a CCD area detector was used to collect 50 161 reflections over 3 d. 6082 unique reflections with I > 3 sigma(I) were used in the multipole model to map the deformation electron density and gave the final statistical factors R(F) = 0.0329, wR(F) = 0.0235 and g.o.f. = 1.37. Structure determination revealed that two O atoms in the crystal structure of the title compound act as hydrogen-bond acceptors for more than one hydrogen bond. Examination of deformation electron density maps showed preferential polarization of the lone-pair electron density of the two O atoms into the shortest hydrogen bonds.


Heterogeneous differentiation of external cells in individual isolated early mouse inner cell masses in culture.
Inner cell masses (ICMs) were isolated from early blastocysts by immunosurgery and incubated in a dense suspension of melanin granules for 3 h after 21 h in culture. The majority of such labelled ICMs subsequently formed outgrowths in vitro in which either giant cells or small solitary cells contained melanin granules. However, a substantial minority produced outgrowths in which both types of cell were unequivocally labelled. Labelled cells appeared from the results of control experiments to have originated within the external layer of the ICM. The giant cells were indistinguishable morphologically from those formed by authentic trophectodermal tissue. The small cells were identified as belonging to the extraembryonic endodermal lineage on the basis of their distribution in host conceptuses following injection into blastocysts. These findings support the conclusion reached in previous studies that early ICM cells can engage in trophectodermal differentiation under certain conditions. In addition, by providing evidence that both trophectoderm and endoderm cells can differentiate from the outer layer of the same ICM, they argue that loss of cellular lability is not coordinated throughout this tissue. Heterogeneity in the differentiation of external cells may depend on differences in both the stage of the mitotic cycle and the number of such cycles that they have completed since fertilization. Finally, cell number in isolated early ICMs was found to increase approximately two-fold during the first 24 h of culture in the present experiments. This contrasts with the results of previous experiments in which cell number either increased more modestly or failed to do so altogether.


Experimental and numerical modelling of the ventriculosinus shunt (El-Shafei shunt).
This study assesses malresorptive hydrocephalus treatment by ventriculosinus shunting with the shunt in the antegrade or retrograde position. First, an experimental model of the cerebral ventricles, the arachnoid villi, the cortical veins, and the superior sagittal sinus was built. For this purpose, the compliance of a human cortical vein was measured and then modelled by means of Penrose tubes. The dimensions of the superior sagittal sinus were determined in vivo by measurements on magnetic resonance imaging scans of 21 patients. Second, a numerical model of the cortical veins and the superior sagittal sinus was built. The numerical results were validated with the results from the experimental model. The experimental and numerical pressure difference between the intracranial pressure and the static sinus pressure was small (0-20 Pa) and corresponded to the theoretically expected values. No overdrainage was found in either the antegrade or the retrograde position of the shunt. Blood reflow was only found while mimicking lumbar puncture or changes in position with the experimental model (lowering the intracranial pressure or increasing the sinus pressure rapidly). Optimal results can be obtained with the shunt positioned in the most downstream half of the superior sagittal sinus. The experimental and numerical results confirm the potential of ventriculosinus shunting as therapy for malresorptive hydrocephalus patients. The ventriculosinus shunt thus proves to be a promising technique.


The (TAAAA)n microsatellite polymorphism in the SHBG gene influences serum SHBG levels in women with polycystic ovary syndrome.
Hyperandrogenaemia is a common feature of polycystic ovary syndrome (PCOS). The sex hormone-binding globulin (SHBG) gene was proposed as being a PCOS candidate gene. A possible influence of the microsatellite polymorphism (TAAAA)(n) in the SHBG gene on serum SHBG levels in PCOS patients was investigated. One hundred and twenty-three PCOS patients and 110 age-matched controls were included in the study. Peripheral blood samples were obtained. Genotyping of the (TAAAA)(n) polymorphism in the SHBG gene was performed. Serum LH, FSH, SHBG and total testosterone concentrations were determined. SHBG alleles with 6-11 TAAAA repeats were found. None of the SHBG alleles or genotypes were present at a significantly more frequent rate in PCOS patients compared with controls. Serum SHBG levels were significantly lower (P < 0.001) in PCOS patients compared with controls and were found to be strongly influenced by the (TAAAA)(n) SHBG polymorphism, in both the PCOS (55.3%) and control (33.1%) groups of patients. The (TAAAA)(n) SHBG gene polymorphism might be an important predictor for serum SHBG levels and, consequently, for hyperandrogenaemic clinical presentation of PCOS.


Substitutions of potentially phosphorylatable serine residues of Bax reveal how they may regulate its interaction with mitochondria.
During apoptosis, the pro-apoptotic protein Bax relocalizes from the cytosol to the mitochondrial outer membrane. This relocalization is associated to major conformational changes, namely at the N- and C-terminal ends of the protein. Substitution of residues located at critical positions within the protein potentially stimulates or inhibits this process. In the present study, we investigated the hypothesis that phosphorylation of serine residues might trigger these conformational changes, with a focus on Ser(163) and Ser(184), which have been shown to be phosphorylatable by protein kinases GSK3beta and Akt/PKB, respectively, and on Ser(60), which is located in a consensus target sequence for PKA. Substitutions of these serine residues by alanine or aspartate were done in wild type or previously characterized Bax mutants, and the capacity of the resulting proteins to interact with mitochondria and to release cytochrome c was assayed in yeast, which provides a tool to study the function of Bax, independently of the rest of the apoptotic network. We conclude that sequential phosphorylation of these serine residues might participate in the triggering of the different conformational changes associated with Bax activation during apoptosis.


Hemi-seesaw nystagmus in lateral medullary syndrome.
Seesaw and hemi-seesaw nystagmus are rare in lateral medullary syndrome. We describe a patient with lateral medullary syndrome who, in addition to other more typical ocular manifestations, demonstrated hemi-seesaw nystagmus (HSSN).


The Hype Surrounding Patent Foramen Ovale Closure and Cryptogenic Stroke: A Walk Through History.
Cryptogenic stroke (CS) represents one-third of all ischaemic strokes. Studies have shown approximately that half of patients with CS have concomitant patent foramen ovale (PFO), with clear data supporting paradoxical embolization as an aetiology of CS. This article is the first of a multi-part review and will detail the history of PFO closure and the clinical trials that have evaluated the efficacy of PFO device closure. Data favour PFO closure in CS for reducing stroke in appropriate patients and should be considered as a treatment modality.


Expression and localization of pannexin-1 and CALHM1 in porcine bladder and their involvement in modulating ATP release.
ATP release from urinary bladder is vital for afferent signaling. The aims of this study were to localize calcium homeostasis modulator 1 (CALHM1) and pannexin-1 expression and to determine their involvement in mediating ATP release in the bladder. To determine gene expression and cellular distribution, PCR and immunohistochemistry were performed, respectively, in the porcine bladder. CALHM1 and pannexin-1-mediated ATP release in response to hypotonic solution (0.45% NaCl)-induced stretch, and extracellular Ca2+ depletion ([Ca2+]0) was measured in isolated urothelial, suburothelial, and detrusor muscle cells. CALHM1 and pannexin-1 mRNA and immunoreactivity were detected in urothelial, suburothelial, and detrusor muscle layers, with the highest expression on urothelium. Hypotonic stretch caused a 2.7-fold rise in ATP release from all three cell populations (P < 0.01), which was significantly attenuated by the pannexin-1 inhibitor, 10Panx1, and by the CALHM1 antibody. Brefeldin A, a vesicular transport inhibitor, and ruthenium red, a nonselective CALHM1 channel blocker, also significantly inhibited stretch-mediated ATP release from urothelial cells. [Ca2+]0 caused a marked, but transient, elevation of extracellular ATP level in all three cell populations. CALHM1 antibody and ruthenium red inhibited [Ca2+]0-induced ATP release from urothelial cells, but their effects on suburothelial and detrusor cells were insignificant. 10Panx1 showed no significant inhibition of [Ca2+]0-induced ATP release in any types of cells. The results presented here provide compelling evidence that pannexin-1 and CALHM1, which are densely expressed in the porcine bladder, function as ATP release channels in response to bladder distension. Modulation of extracellular Ca2+ may also regulate ATP release in the porcine bladder through voltage-gated CALHM1 ion channels.


Neurotoxicity of Micrurus altirostris (Uruguayan coral snake) venom and its neutralization by commercial coral snake antivenom and specific antiserum raised in rabbits.
In this work, we studied the neuromuscular blockade caused by Micrurus altirostris venom (0.1-10 microg/mL) in indirect stimulated chick biventer cervicis and mouse phrenic nerve-diaphragm preparations and the ability of commercial antivenom (Instituto Butantan) and antiserum raised in rabbits to neutralize neurotoxicity and lethality in chicks and mice (LD(50) 0.042 and 0.255 mg/kg), injected i.m. and i.p., respectively, with venom (5 LD(50)):antivenom or antiserum mixtures (n = 6) of 1:1-1:2.5-1:5-1:10-1:20. The venom caused a complete and irreversible neuromuscular blockade in both preparations, inhibited the acetylcholine and carbachol contractures, without interfering on KCl response. The neuromuscular blockade was not Ca(2+) or temperature-dependent and did not affect the response to direct stimulation. Only a venom:antivenom or antiserum ratio of 1:20 neutralized the neuromuscular blockade in vitro and protected chicks and mice against 5 LD(50) of venom. Our results indicated that Micrurus altirostris venom interferes with postsynaptic neurotransmission and that commercial antivenom and rabbit antiserum have low efficacy in neutralizing the neurotoxicity and lethality of this venom.


Improved effect of amikacin-loaded poly(D,L-lactide-co-glycolide) nanoparticles against planktonic and biofilm cells of Pseudomonas aeruginosa.
Amikacin is one of the most effective antibiotics against Pseudomonas aeruginosa infections, but because of its high toxicity, the use of this antibiotic has been clinically limited. In the present study, amikacin was successfully loaded into a new formulation of nanoparticles (NPs) based on poly(d,l-lactide-co-glycolide) 50 : 50 in order to enhance the treatment efficacy. The synthetized amikacin-loaded PLGA nanoparticles with high drug loading and stability were used to eliminate P. aeruginosa cells in planktonic and biofilm states. P. aeruginosa PAO1 biofilm susceptibility studies were done using the minimum biofilm eradication concentration assay. The association of fluorescently labeled amikacin-loaded nanoparticles (A-NPs) with mouse monocyte macrophage cells (RAW 264.7), and the nanoparticles ability to interact and eradicate the bacterial cells even in the form of biofilms, was investigated using Flow cytometric studies and confocal laser scanning microscopy. Flow cytometric studies showed that these NPs were able to interact with planktonic and biofilm bacterial cells. Moreover, following 1 h of incubation of A-NPs with 1-day-old biofilm, it was found that particles penetrate through the entire biofilm thickness. Live/dead fluorescent staining followed by CLSM analysis showed that the A-NPs were more effective than free drug in biofilm eradication. The good antibacterial and antibiofilm activities of A-NPs, in addition to their ability to enter macrophages without any cytotoxicity for these cells, make them a potential candidate to treat P. aeruginosa infections.


Functional role of calcium signals for microglial function.
In this review we summarize mechanisms of Ca(2+) signaling in microglial cells and the impact of Ca(2+) signaling and Ca(2+) levels on microglial function. So far, Ca(2+) signaling has been only characterized in cultured microglia and thus these data refer rather to activated microglia as observed in pathology when compared with the resting form found under physiological conditions. Purinergic receptors are the most prominently expressed ligand-gated Ca(2+)-permeable channels in microglia and control several microglial functions such as cytokine release in a Ca(2+)-dependent fashion. A large variety of metabotropic receptors are linked to Ca(2+) release from intracellular stores. Depletion of these intracellular stores triggers a capacitative Ca(2+) entry. While microglia are already in an activated state in culture, they can be further activated, for example, by exposure to bacterial endotoxin. This activation leads to a chronic increase of [Ca(2+)](i) and this Ca(2+) increase is a prerequisite for the release of nitric oxide and cytokines. Moreover, several factors (TNFalpha, IL-1beta, and IFN-gamma) regulate resting [Ca(2+)](i) levels.