Increased expression of urotensin II and urotensin II receptor in human diabetic nephropathy.
Urotensin II (UII) is an 11-amino acid vasoactive peptide, recently identified as the ligand for a novel G protein-coupled receptor, GPR-14 (renamed urotensin receptor [UT]). In addition to its potent vasoconstrictive actions, UII also has trophic and profibrotic effects, leading to its implication in the pathogenesis of heart failure. However, elevated plasma UII levels also were reported in association with renal impairment and diabetes. Accordingly, the present study sought to examine the expression and localization of UII and its receptor in kidney tissue from patients with diabetic nephropathy. We quantified UII and UT gene expression in renal biopsy tissue samples from patients with diabetic nephropathy by using quantitative real-time polymerase chain reaction and determined the intrarenal distribution of their peptides by means of immunohistochemistry. In human diabetic tissue, gene expression of UII and UT were increased 45- and almost 2,000-fold in comparison to control nephrectomy tissue, respectively (P < 0.0001). Immunohistochemical studies showed intense UII peptide staining in diabetic tissue localized predominantly to tubular epithelial cells, and fluorescein-labeled ligand binding studies showed a similar tubular pattern of distribution. In the context of its known biological actions, the dramatic overexpression of UII and its receptor implicate this vasoactive peptide as a possible novel factor in the pathogenesis of diabetic nephropathy.


[Pharmacotherapy follow-up for patients admitted to the Internal Medicine Department of Hospital Infanta Margarita].
In pharmacotherapeutic follow-up a pharmacist is responsible for drug-related patient needs (DRPN) by detecting, preventing and solving medication-related problems aiming at specific results to improve patient quality of life. Drug-related problems are pharmacotherapy failures leading to failed therapeutic goals or undesirable events. In this study, Daders methodology for pharmacotherapeutic follow-up was used in patients admitted to the Internal Medicine Department of Hospital Infanta Margarita, Cabra-Córdoba, Spain. In all, 85 DRPNs (2.7 DRPNs per patient) were identified, and 36 pharmaceutical procedures were performed, with physicians accepting 92% of said procedures. Forty-nine percent of drug-related problems were related to need, 40% to effectiveness, and 11% to safety. The presence of a pharmacist at the Internal Medicine Department allows the detection of DRPNs that are mostly related to need and effectiveness. Pharmaceutical procedures are widely accepted by medical teams.


A cautionary note: Toxicity of polyethylene glycol 200 injected intraperitoneally into mice.
The parenteral administration of hydrophobic substances in vivo requires the use of organic solvents to ensure sufficient solubility and avoid precipitation. Dimethyl sulfoxide is commonly used for this purpose. Based on the common assumption that polyethylene glycol (PEG) is non-toxic, our local regulatory authorities recently recommended the use of PEG instead. However, mice injected intraperitoneally (i.p.) with PEG 200 at a dose of 8 mL/kg (i.e. 9 g/kg) did not tolerate PEG 200 well, and half of the animals had to be euthanized. Our results demonstrate that although PEG 200 is generally considered to be harmless, it can be toxic when injected i.p. and is painful for the recipient mice. Nevertheless, it can be used as a solvent for repeated i.p. injections in mice at a dose of 2 mL/kg (i.e. 2.25 g/kg) without obvious signs of systemic toxicity.


Impact of supine versus prone position on endoscopic retrograde cholangiopancreatography performance: a retrospective study.
Endoscopic retrograde cholangiopancreatography (ERCP) is frequently performed in the prone or supine position. We compared the technical success and other outcomes between these positions. This was a retrospective cohort study using the Clinical Outcomes Research Initiative database. Demographics, procedure and fluoroscopy time, visualization of main structures, and technical success rates were compared between the supine and prone positions. Univariate and multivariate regressions were performed to adjust for age, sex, ethnicity and clinical setting. A total of 21,090 patients who underwent ERCP were included, of whom 1769 (8.4%) were supine and 19,321 (91.6%) were prone. The common bile duct (CBD) was visualized and cannulated in 89.1% of supine vs. 91.4% of prone positions (P=0.017), while the ampulla was visualized in 97.1% of supine vs. 97.7% of prone (P=0.414). The ERCP was incomplete in 10% of supine vs. 5% of prone cases (P<0.001). On multivariate analysis, supine position required shorter procedure times than prone (adjusted odds ratio [aOR] 0.98, 95% confidence interval [CI] 0.98-0.98; P<0.001). The supine position also yielded lower odds of CBD visualization and cannulation (aOR 0.63, 95%CI 0.44-0.91; P=0.011) and higher odds of an incomplete examination (aOR 1.84, 95%CI 1.46-2.30; P<0.001) vs. prone. The supine position leads to shorter procedures but is more likely to result in poorer visualization and cannulation of the CBD and an incomplete examination. This may reflect the technical difficulty of performing ERCP in the supine position for the endoscopist. Our study supports recommendations for an individualized ERCP approach.


Effect of excitotoxic lesions of rat medial prefrontal cortex on spatial memory.
The involvement of medial prefrontal cortex (mPFC) in spatial learning was examined in two memory tasks using spatial components, the Morris water maze and the three-panel runway. Using the Morris water maze task, with an invisible platform, the effects of NMDA mPFC lesions were assessed in a procedure reflecting spatial learning and memory, including a spatial reversal. In the three-panel runway, a delayed matching-to-position procedure was used in which rats were required to find food at the end of the runway after passing through one of three panel gates set into four barriers spaced equally apart along the maze. In addition, mPFC lesions were assessed behaviorally in two behavioral tests known to be sensitive to mPFC dysfunction: the food hoarding paradigm and spontaneous locomotion in the open field. Consistent with the documented effects of mPFC damage, NMDA mPFC lesions impaired food hoarding behavior and increased spontaneous exploratory locomotion. In the Morris water maze and the three-panel runway, mPFC-lesioned rats showed relatively few effects, supporting the conclusion that the damage inflicted to the mPFC had no consequence for the processing of spatial information. However, mPFC lesioned animals showed slower acquisition during both the training trial in the three-panel runway and the reversal training in the Morris water maze. These results suggest that spatial memory did not depend on mPFC integrity in the Morris water maze and the three-panel runway experiments, and address the issue of deficits induced by mPFC lesions in memory tasks dependent on non-mnemonic processes such as attentional processes and/or a reduced behavioral flexibility to environmental changes.


Activated Sludge and other Aerobic Suspended Culture Processes.
This is a literature review for the year 2015 and contains information specifically associated with suspended growth processes including activated sludge, upflow anaerobic sludge blanket, and sequencing batch reactors. The review encompasses modeling and kinetics, nutrient removal, system design and operation. Compared to past reviews, many topics show increase in activity in 2015. These include, fate and effect of xenobiotics, industrial wastes treatment with sludge, and pretreatment for the activated sludge. These topics are referred to the degradation of constituents in activated sludge. Other sections include population dynamics, process microbiology give an insight into the activated sludge. The subsection in industrial wastes: converting sewage sludge into biogases was also mentioned.


The heat is on: protecting your patients from nature's silent killer.
Hot weather may be uncomfortable, but for many, it can be a killer. This article describes different types of heat-related illness, risk factors for heat stroke, and outlines targeted nursing interventions and critical environmental assessments. Nurses can use this information to identify at-risk patients and conduct patient and family teaching. Because home care and hospice nurses and other staff often work in severe heat, this knowledge is also important for personal health and safety.


Characterization and Biosynthesis of a Rare Fungal Hopane-Type Triterpenoid Glycoside Involved in the Antistress Property of Aspergillus fumigatus.
A rare fungal hopane-type triterpenoid glycoside fumihopaside A was identified by genome mining in combination with HPLC-MS/MS in Aspergillus fumigatus. Combining genetic deletions in A. fumigatus with heterologous reconstitutions in Aspergillus nidulans of the afum gene cluster, we identified one fungal squalene hopane cyclase AfumA charging the cyclization of the hopene skeleton, one cytochrome P450, and one UDP-glycosyltransferase. Bioassays indicated that fumihopaside A plays important roles in protecting A. fumigatus against heat or ultraviolet stress.


Role of Adaptive Immune and Impacts of Risk Factors on Adaptive Immune in Alzheimer's Disease: Are Immunotherapies Effective or Off-Target?
The pathogenesis of Alzheimer's disease (AD) is complex. Still it remains unclear, which resulted in all efforts for AD treatments with targeting the pathogenic factors unsuccessful over past decades. It has been evidenced that the innate immune is strongly implicated in the pathogenesis of AD. However, the role of adaptive immune in AD remains mostly unknown and the results obtained were controversial. In the review, we summarized recent studies and showed that the molecular and cellular alterations in AD patients and its animal models involving T cells and B cells as well as immune mediators of adaptive immune occur not only in the peripheral blood but also in the brain and the cerebrospinal fluid. The risk factors that cause AD contribute to AD progress by affecting the adaptive immune, indicating that adaptive immunity proposes a pivotal role in this disease. It may provide a possible basis for applying immunotherapy in AD and further investigates whether the immunotherapies are effective or off-target?


Thyroid uptake neck phantoms are not created equal.
The main purpose of this communication is to alert nuclear medicine departments to the fact that the earlier version of the water phantom grossly overestimates soft-tissue attenuation in the neck, resulting in calculated thyroid uptake values which are significantly overestimated (in hyperthyroid patients we noted uptake values approaching or exceeding 100%). We believe that the solid Lucite phantom (which is the one recommended by IAEA) better approximates the human neck soft tissue overlying the thyroid. Institutions that continue to use the water phantom should be aware that their thyroid uptakes will be relatively elevated and the normal range must be shifted accordingly. Our normal range is 10%-30% uptake of 123I at 24 hr for the Lucite phantom. For the water phantom, the estimated normal range would be 15%-45%. In addition, the phantom type should be considered when comparing uptake results with those from another institution for a particular patient. Also, treatment doses for Grave's disease could be significantly affected, if such doses are calculated by a formula that depends on uptake.


TBC1D3 promotes neural progenitor proliferation by suppressing the histone methyltransferase G9a.
Genomic changes during human linage evolution contribute to the expansion of the cerebral cortex to allow more advanced thought processes. The hominoid-specific gene TBC1D3 displays robust capacity of promoting the generation and proliferation of neural progenitors (NPs), which are thought to contribute to cortical expansion. However, the underlying mechanisms remain unclear. Here, we found that TBC1D3 interacts with G9a, a euchromatic histone lysine N-methyltransferase, which mediates dimethylation of histone 3 in lysine 9 (H3K9me2), a suppressive mark for gene expression. TBC1D3 displayed an inhibitory role in G9a's histone methyltransferase activity. Treatment with G9a inhibitor markedly increased NP proliferation and promoted human cerebral organoid expansion, mimicking the effects caused by TBC1D3 up-regulation. By contrast, blockade of TBC1D3/G9a interaction to disinhibit G9a caused up-regulation of H3K9me2, suppressed NP proliferation, and impaired organoid development. Together, this study has demonstrated a mechanism underlying the role of a hominoid-specific gene in promoting cortical expansion.


[Pathology of basal ganglia in neurodegenerative diseases].
Intra- and/or extracellular proteinaceous inclusions in the brain tissue are characteristic pathological markers of many neurodegenerative diseases. Tau protein in neurofibrillary tangles and beta-amyloid in senile plaques are associated with Alzheimer's disease. Tau is associated with various neurological conditions, which are collectively referred to as tauopathies. Alpha-synucleinopathy is a term that collectively refers to a set of diseases in which neurodegeneration is accompanied by intracellular accumulation of alpha-synuclein in neurons or glial cells. Recently, TDP-43 has been identified as a major disease protein in the ubiquitinated inclusions in deseases such as amyotrophic lateral sclerosis and frontotemporal lobar degeneration with tau-negative, ubiquitin-positive inclusions. Thus, these neurodegenerative disorders comprise a new disease class, namely, TDP-43 proteinopathy. In this article, we review the present understanding of histopathological features of basal ganglia lesions in protein conformation disorders, including tauopathy, alpha-synucleinopathy, and TDP-43 proteinopathy.


Glucocorticoid Receptor-Tethered Mineralocorticoid Receptors Increase Glucocorticoid-Induced Transcriptional Responses.
Mineralocorticoid and glucocorticoid receptors (MRs and GRs) constitute a functionally important dual receptor system detecting and transmitting circulating corticosteroid signals. High expression of MRs and GRs occurs in the same cells in the limbic system, the primary site of glucocorticoid action on cognition, behavior, and mood; however, modes of interaction between the receptors are poorly characterized. We used chromatin immunoprecipitation with nucleotide resolution using exonuclease digestion, unique barcode, and single ligation (ChIP-nexus) for high-resolution genome-wide characterization of MR and GR DNA binding profiles in neuroblastoma cells and demonstrate recruitment to highly similar DNA binding sites. Expressed MR or GR showed differential regulation of endogenous gene targets, including Syt2 and Ddc, whereas coexpression produced augmented transcriptional responses even when MRs were unable to bind DNA (MR-XDBD). ChIP confirmed that MR-XDBD could be tethered to chromatin by GR. Our data demonstrate that MR can interact at individual genomic DNA sites in multiple modes and suggest a role for MR in increasing the transcriptional response to glucocorticoids.


Estradiol regulation of P-glycoprotein expression in mouse kidney and human tubular epithelial cells, implication for renal clearance of drugs.
P-glycoprotein (P-gp/ABCB1) is an ATP-binding cassette drug efflux transporter expressed in a variety of tissues that affects the pharmacokinetic disposition of many drugs. Although several studies have reported gender-dependent differences in the expression of P-gp, the role of sex hormones in regulating the expression of P-gp and its transport activity has not been well understood. In this study, we demonstrated that 17β-estradiol has the ability to induce the expression of P-pg in mouse kidneys and cultured human renal proximal tubular epithelial cells. After intravenous injection of a typical P-gp substrate, digoxin, renal clearance in female mice was approximately 2-fold higher than that in male mice. The expression of murine P-gp and its mRNA (Abcb1a and Abcb1b) were also higher in female mice than in male mice. The expression of P-gp in cultured renal tissues prepared from female and male mice was significantly increased by 17β-estradiol, but not testosterone. Similar 17β-estradiol-induced expression of P-gp was also detected in cultured human tubular epithelial cells, accompanied by the enhancement of its transport activity of digoxin. The present findings suggest the contribution of estradiol to female-predominant expression of P-gp in renal cells, which is associated with sex-related disparities in the renal elimination of digoxin.


Contribution of BubR1 to oxidative stress-induced aneuploidy in p53-deficient cells.
DNA aneuploidy is observed in various human tumors and is associated with the abnormal expression of spindle assembly checkpoint (SAC) proteins. Oxidative stress (OS) causes DNA damage and chromosome instability that may lead to carcinogenesis. OS is also suggested to contribute to an increase in aneuploid cells. However, it is not clear how OS is involved in the regulation of SAC and contributes to carcinogenesis associated with aneuploidy. Here we show that an oxidant (KBrO3) activated the p53 signaling pathway and suppressed the expression of SAC factors, BubR1, and Mad2, in human diploid fibroblast MRC5 cells. This suppression was dependent on functional p53 and reactive oxygen species. In p53 knockdown cells, KBrO3 did not suppress BubR1 and Mad2 expression and increased both binucleated cells and cells with >4N DNA content. BubR1 and not Mad2 downregulation suppressed KBrO3-induced binucleated cells and cells with >4N DNA content in p53 knockdown cells, suggesting that BubR1 contributes to enhanced polyploidization by a mechanism other than its SAC function. In analysis of 182 gastric cancer specimens, we found that BubR1 expression was significantly high when p53 was positively stained, which indicates loss of p53 function (P = 0.0019). Moreover, positive staining of p53 and high expression of BubR1 in tumors were significantly correlated with DNA aneuploidy (P = 0.0065). These observations suggest that p53 deficiency may lead to the failure of BubR1 downregulation by OS and that p53 deficiency and BubR1 accumulation could contribute to gastric carcinogenesis associated with aneuploidy.


Polymerase Chain Reaction-Based Malaria Diagnosis Can Be Increasingly Adopted during Current Phase of Malaria Elimination in India.
Despite commendable progress in the control of malaria in India and other countries, there are hidden reservoirs of parasites in human hosts that continually feed malaria transmission. Submicroscopic infections are known to be a significant proportion in low-endemic settings like India and these infections do possess transmission potential. Hence, these reservoirs of infection add to the existing roadblocks for malaria elimination. It is crucial that this submerged burden of malaria is detected and treated to curtail further transmission. The currently used diagnostic tools including the so-called "gold standard" of microscopy are incapable of detecting these submicroscopic infections and thus are suboptimal. It is an opportune time to usher in more sensitive molecular tools like polymerase chain reaction (PCR) for routine diagnosis at all levels of healthcare as an additional diagnostic tool in routine settings. Polymerase chain reaction assays have been developed into user-friendly formats for field diagnostics and are near point of care. In India, because of the COVID-19 pandemic, these are being used rampantly across the country. The facilities created for COVID-19 diagnosis can easily be co-opted and harnessed for malaria diagnosis to augment surveillance by the inclusion of molecular techniques like PCR in the routine national malaria control program.


On a calculus-based statistics course for life science students.
The choice of pedagogy in statistics should take advantage of the quantitative capabilities and scientific background of the students. In this article, we propose a model for a statistics course that assumes student competency in calculus and a broadening knowledge in biology. We illustrate our methods and practices through examples from the curriculum.


CAR T Cell Therapy in Primary Brain Tumors: Current Investigations and the Future.
Chimeric antigen receptor T cells (CAR T cells) are engineered cells expressing a chimeric antigen receptor (CAR) against a specific tumor antigen (TA) that allows for the identification and elimination of cancer cells. The remarkable clinical effect seen with CAR T cell therapies against hematological malignancies have attracted interest in developing such therapies for solid tumors, including brain tumors. Glioblastoma (GBM) is the most common primary brain tumor in adults and is associated with poor prognosis due to its highly aggressive nature. Pediatric brain cancers are similarly aggressive and thus are a major cause of pediatric cancer-related death. CAR T cell therapy represents a promising avenue for therapy against these malignancies. Several specific TAs, such as EGFR/EGFRvIII, IL13Rα2, B7-H3, and HER2, have been targeted in preclinical studies and clinical trials. Unfortunately, CAR T cells against brain tumors have showed limited efficacy due to TA heterogeneity, difficulty trafficking from blood to tumor sites, and the immunosuppressive tumor microenvironment. Here, we review current CAR T cell approaches in treating cancers, with particular focus on brain cancers. We also describe a novel technique of focused ultrasound controlling the activation of engineered CAR T cells to achieve the safer cell therapies. Finally, we summarize the development of combinational strategies to improve the efficacy and overcome historical limitations of CAR T cell therapy.


Effects of Feeding Frequency Using a Commercial Automated Feeding Device on Gastric Ulceration in Exercised Quarter Horses.
The objective of this study was to evaluate two feeding regimens: traditional grain feeding (twice a day, traditional fed [TF]) compared with horses fed with an automated feeder programmed to deliver grain in 20 equal aliquots (fractioned fed [FF]) and its effects on gastric ulceration, body weight, and body condition score (BCS). Thirty-one Quarter Horses were maintained in individual stalls during the 60-day experimental period and worked at moderate exercise intensity. Gastroscopies were performed before grain adaptation at 0, 30, and 60 days. Weight and BCS were evaluated biweekly. There were no changes in squamous (equine squamous gastric disease [ESGD]) ulcers in FF horses over the study period and higher ESGD ulcer scores in TF horses over both the 30 and 60-day time points. The proportion of horses with equine glandular gastric disease (EGGD) were significantly lower with fractioned feed when compared with traditional feed on day 30, but no difference was seen at day 60. No significant differences were found when comparing the two groups for BCS or body weight throughout the study period. This study further supports that fasting period is a strong contributing factor for gastric ulcer formation. FF of the daily total grain led to a decrease in time fasting when compared with TF which reduced the severity and incidence of both ESGD and EGGD. The use of the commercial feeder for FF may aid in lessening prevalence of gastric ulcers in young training Quarter Horses.


Structure-activity relationship in the auxin activity of mono-substituted phenylacetic acids.
The analysis of substituent constants for the lipophilic and electronic factors in the auxin activity of substituted phenylacetic acids in elongation of coleoptile segments shows that these factors parallel those for the phenoxyacetic acids but assign reactivity in growth promotion to the meta position of phenylacetic acid. The inhibitory effects with supra-optimal concentrations are highly dependent on the lipophilic character of the molecules.


Comparative activity of cefepime, alone and in combination, against clinical isolates of Pseudomonas aeruginosa and Pseudomonas cepacia from cystic fibrosis patients.
The comparative in vitro activity and synergy of cefepime were evaluated with clinical isolates of Pseudomonas aeruginosa and Pseudomonas cepacia from cystic fibrosis patients. The activity of cefepime, both alone and in combination, was comparable to those of other antibiotics. The clinical efficacy of cefepime in cystic fibrosis patients merits investigation.


Enhancement of periodontal tissue regeneration by transplantation of bone marrow mesenchymal stem cells.
The use of suitable cells transplanted into periodontal osseous defects appears to be a powerful strategy to promote periodontal tissue regeneration. Mesenchymal stem cells (MSCs) isolated from bone marrow have the potential for multilineage differentiation. The purpose of this study was to examine whether auto-transplantation of MSCs into periodontal osseous defects would be useful for periodontal tissue regeneration. Bone marrow MSCs were isolated from beagle dogs and expanded in vitro. The expanded MSCs were mixed with atelocollagen (2% type I collagen) at final concentrations of 2 x 10(6), 5 x 10(6), 1 x 10(7), or 2 x 10(7) cells/ml, and auto-transplanted into experimental Class III defects. Atelocollagen alone was implanted into the defects as a control. Periodontal tissue healing was evaluated by histological and morphometric analyses 1 month after transplantation. The defects were regenerated with cementum, periodontal ligament, and alveolar bone in the MSC-atelocollagen groups. Less periodontal tissue regeneration was observed in the control group compared to the MSC-atelocollagen groups. Morphometric analysis revealed that the percentage of new cementum length in the 5 x 10(6) and 2 x 10(7) cells/ml groups and the percentage of new bone area in the 2 x 10(7) cells/ml group were significantly higher than in the control group (P < 0.01). These findings suggest that auto-transplantation of bone marrow mesenchymal stem cells is a novel option for periodontal tissue regeneration.


Total Robotic Myomectomy in a Complex Case: Extending the Limits.
We present the case of a 30-year-old female with primary infertility who had multiple large myomas up to 22 weeks uterine size. She had a body mass index of 42 kg/m2 and also concurrent endometriosis, for which she was on medical management. After meticulous preoperative planning, total robotic myomectomy and endometriosis clearance was done. Four large myomas weighing a total of 750 g were removed after morcellation. Console time was 160 min, and she made an uneventful recovery with only 0.1 g/dL drop in hemoglobin. Robotic myomectomy is considered as an improvement over laparotomy for patients with up to three myomas and when the surgical time does not exceed 4 hours. We were successful in our attempt at total robotic myomectomy to extend these limits and had a positive surgical outcome.


A Multiscale Mathematical Model of Tumour Invasive Growth.
Known as one of the hallmarks of cancer (Hanahan and Weinberg in Cell 100:57-70, 2000) cancer cell invasion of human body tissue is a complicated spatio-temporal multiscale process which enables a localised solid tumour to transform into a systemic, metastatic and fatal disease. This process explores and takes advantage of the reciprocal relation that solid tumours establish with the extracellular matrix (ECM) components and other multiple distinct cell types from the surrounding microenvironment. Through the secretion of various proteolytic enzymes such as matrix metalloproteinases or the urokinase plasminogen activator (uPA), the cancer cell population alters the configuration of the surrounding ECM composition and overcomes the physical barriers to ultimately achieve local cancer spread into the surrounding tissue. The active interplay between the tissue-scale tumour dynamics and the molecular mechanics of the involved proteolytic enzymes at the cell scale underlines the biologically multiscale character of invasion and raises the challenge of modelling this process with an appropriate multiscale approach. In this paper, we present a new two-scale moving boundary model of cancer invasion that explores the tissue-scale tumour dynamics in conjunction with the molecular dynamics of the urokinase plasminogen activation system. Building on the multiscale moving boundary method proposed in Trucu et al. (Multiscale Model Simul 11(1):309-335, 2013), the modelling that we propose here allows us to study the changes in tissue-scale tumour morphology caused by the cell-scale uPA microdynamics occurring along the invasive edge of the tumour. Our computational simulation results demonstrate a range of heterogeneous dynamics which are qualitatively similar to the invasive growth patterns observed in a number of different types of cancer, such as the tumour infiltrative growth patterns discussed in Ito et al. (J Gastroenterol 47:1279-1289, 2012).


Who failed to enroll in Medicare Part D, and why? Early results.
Early results on the Medicare Part D prescription drug program, from a survey of people age sixty-five and older who were interviewed just before enrollment started and just after it ended, indicate that Medicare has met its target of 90 percent coverage. Enrollment rates in vulnerable subpopulations-poor health, low income, or cognitive impairment-are almost high enough to offset lower rates of other coverage. However, sizable numbers of elderly people remain uncovered, contrary to their self-interest. Seniors give Part D mixed reviews, and majorities are less satisfied with Medicare and with the government as a result of their experience with this program.


Implementing a health promotion strategy in occupational therapy education and practice.
In response to the growing emphasis on health promotion, this article addresses four major questions: 1) How is occupational therapy's client-centered philosophy reflected in the principles outlined by the World Health Organization (1984)? 2) What are the roles of occupational therapy in health promotion? 3) How will occupational therapy education foster this development? 4) How will the practice of occupational therapy respond to the challenges and implement the strategies outlined in "Achieving Health for All: A Framework for Health Promotion"? Despite similarities between occupational therapy's client-centered approach and the health promotion framework, the need for occupational therapy education to reflect the developments in health promotion and prevention is very evident. A four part model is proposed, and the implications for graduate education as well as continuing professional education are outlined. Recommendations for practice, education, research and development are discussed.


[Optimization of the immunoelectrophoresis technic (western blot) for the confirmation of human immunodeficiency virus infection (HIV) in Panama].
The purpose of this study is to report the results of the authors' investigation to apply the western blot technique (WB UP-LCS) in the diagnosis of human immunodeficiency virus type 1 (HIV-1) infection. To do this, the authors separated the proteins of the HIV-1 virus by electrophoresis, based on their molecular weight, in poliacilamide gel with SDS (SDS-PAGE) during 3 hours at 200 volts. Then they electrotransferred these proteins to nitrocellulose paper during four hours at 200 milliamperes, with the aid of external cooling. The nitrocellulose strips were evaluated considering the incubation time (1 and 16 hours), two conjugates (human anti IgG with Peroxidase and human anti IgG Biotin plus Streptatividine with Peroxidase) and two dilutions of the patients' sera (1/50 and 1/100). Based on their results the Authors conclude that, in the first place, the optimal conditions for the test include a dilution of 1/100 of the patients serum, incubation of the serum for 16 hours and the use of the conjugate of anti human IgG with Biotin and Streptavidine with Peroxidase; secondary, that the immunologic reactivity against proteins p24 and gp 160/120 is the most important diagnostic criterion for the confirmation of infection with HIV-1 and that they obtained a diagnostic correlation of 100% at a cost which was 5 to 7 times less than that of the commercial system.


Changes in the molecular packing of fibrillin microfibrils during extension indicate intrafibrillar and interfibrillar reorganization in elastic response.
Fibrillin-rich microfibrils are the major structural components of the extracellular matrix that provide elasticity in a majority of connective tissues. The basis of elastic properties lies in the organization of fibrillin molecules, which, unfortunately, is still poorly understood. An X-ray diffraction study of hydrated fibrillin-rich microfibrils from zonular filaments has been conducted to give an insight into the molecular structure of microfibrils in intact tissue. A series of measurements was taken during controlled tissue extension to observe alterations in the lateral packing of microfibrils. Computer-generated simulated patterns were used to fit the experimental X-ray scattering data and to obtain the fibril diameter and lateral distance between the fibrils. The results suggest a nonlinear correlation between external strain and decrease in fibril diameter and lateral spacing. This was accompanied by a nonlinear increase in axial periodicity and a structure with a 160-nm periodicity, which is reported here for the first time using X-ray diffraction. These changes may reflect the unraveling of fibrillin from the complex folded arrangement into a linear structure. This finding supports a pleating model where fibrillin molecules are highly folded within the microfibrils; more importantly, the connection is made between the interaction of individual microfibrils and the change in their suprafibrillar coherent organization during extension. We suggest that the intermediate states observed in our study reflect sequential unfolding of fibrillin and can explain the process of its reversible unraveling.


A randomized clinical trial on the effects of remote intercessory prayer in the adverse outcomes of pregnancies.
The scope of this article was to investigate whether intercessory prayer (IP) influences the adverse outcomes of pregnancies. A double-blind, randomized clinical trial was conducted with 564 pregnant women attending a prenatal public health care service. The women were randomly assigned to an IP group or to a control group (n = 289 per group). They were simultaneously and randomly assigned to practice prayer off-site or not. The following parameters were evaluated: Apgar scores, type of delivery and birth weight. The mean age of the women was 25.1 years of age (± 7.4), and the average gestational age was 23.4 weeks (± 8.1). The average number of years of schooling for the women was 8.1 years (± 3.1). The women in the IP and control groups presented a similar number of adverse medical events with non-significant p. No significant differences were detected in the frequency of adverse outcomes in pregnant women who practiced IP and those in the control group.


Symptoms of posttraumatic stress disorder and depression among children in tsunami-affected areas in southern Thailand.
On December 26, 2004, an undersea earthquake occurred off the northwestern coast of Sumatra, Indonesia. The tsunami that followed severely impacted all 6 southwestern provinces of Thailand, where approximately 20,000 children were directly affected. To assess trauma experiences and the prevalence of symptoms of posttraumatic stress disorder (PTSD) and depression among children in tsunami-affected provinces in southern Thailand. Population-based mental health surveys were conducted among children aged 7 to 14 years in Phang Nga, Phuket, and Krabi provinces from February 15-22, 2005 (2 months posttsunami), and September 7-12, 2005 (9 months posttsunami). Trauma experiences and symptoms of PTSD and depression as measured by a tsunami-modified version of the PsySTART Rapid Triage System, the UCLA PTSD Reaction Index, and the Birleson Depression Self-Rating Scale. A total of 371 children (167 displaced and living in camps, 99 not displaced from villages affected by the tsunami, and 105 not displaced from unaffected villages) participated in the first survey. The prevalence rates of PTSD symptoms were 13% among children living in camps, 11% among children from affected villages, and 6% among children from unaffected villages (camps vs unaffected villages, P = .25); for depression symptoms, the prevalence rates were 11%, 5%, and 8%, respectively (P = .39). In multivariate analysis of the first assessment, having had a delayed evacuation, having felt one's own or a family member's life to have been in danger, and having felt extreme panic or fear were significantly associated with PTSD symptoms. Older age and having felt that their own or a family member's life had been in danger were significantly associated with depression symptoms. In the follow-up survey, 72% (151/210) of children from Phang Nga participated. Prevalence rates of symptoms of PTSD and depression among these children did not decrease significantly over time. This assessment documents the prevalence of mental health problems among children in tsunami-affected provinces in southern Thailand at 2 and 9 months posttsunami. Traumatic events experienced during the tsunami were significantly associated with symptoms of PTSD and depression. These data may be useful to target mental health services for children and may inform the design of these interventions.


Strength of the pnicogen bond in complexes involving group Va elements N, P, and As.
A set of 36 pnicogen homo- and heterodimers, R3E···ER3 and R3E···E′R′3, involving differently substituted group Va elements E = N, P, and As has been investigated at the ωB97X-D/aug-cc-pVTZ level of theory to determine the strength of the pnicogen bond with the help of the local E···E′ stretching force constants k(a). The latter are directly related to the amount of charge transferred from an E donor lone pair orbital to an E′ acceptor σ* orbital, in the sense of a through-space anomeric effect. This leads to a buildup of electron density in the intermonomer region and a distinct pnicogen bond strength order quantitatively assessed via k(a). However, the complex binding energy ΔE depends only partly on the pnicogen bond strength as H,E-attractions, H-bonding, dipole-dipole, or multipole-multipole attractions also contribute to the stability of pnicogen bonded dimers. A variation from through-space anomeric to second order hyperonjugative, and skewed π,π interactions is observed. Charge transfer into a π* substituent orbital of the acceptor increases the absolute value of ΔE by electrostatic effects but has a smaller impact on the pnicogen bond strength. A set of 10 dimers obtains its stability from covalent pnicogen bonding whereas all other dimers are stabilized by electrostatic interactions. The latter are quantified by the magnitude of the local intermonomer bending force constants XE···E′. Analysis of the frontier orbitals of monomer and dimer in connection with the investigation of electron difference densities, and atomic charges lead to a simple rationalization of the various facets of pnicogen bonding. The temperature at which a given dimer is observable under experimental conditions is provided.


Memory for everyday information in students with learning disabilities.
This study compared students with and without learning disabilities (LD) on their recall of academic information and information encountered in the students' everyday lives. The academic recall measures included a sentence listening span test, a rhyming words working memory test, and a visual matrix working memory task. Students' cued recall of all the tasks was also measured. The everyday working memory tasks included a dance episode event recall test; a library procedure recall test; and recall tests of commonly found objects, such as a coin, a telephone, and a McDonald's sign. Compared to students without LD, students with LD performed poorly on both the academic recall tasks and the everyday recall tasks. These results support the notion that some students with LD may have working memory problems that affect their performance on tasks other than reading. The results of the cued recall showed that the availability of cues significantly decreased the ability group differences on many of the academic and everyday tasks. This result replicates prior research findings that students with LD do not use retrieval strategies effectively and that some students with LD may have a production deficiency that affects their retrieval of previously encoded information.


Signals that initiate, augment, and provide directionality for human keratinocyte motility.
Human keratinocytes (HK) migration plays a critical role in the re-epithelialization of acute skin wounds. Although extracellular matrices (ECM) and growth factors (GF) are the two major pro-motility signals, their functional relationship remains unclear. We investigated how ECM and GF regulate HK motility under defined conditions: (1) in the absence of GF and ECM and (2) with or without GF with cells apposed to a known pro-motility ECM. Our results show that HK migrate on selected ECM even in the total absence of GF. This suggests that certain ECM alone are able to "initiate" HK migration. Unlike ECM, however, GF alone cannot initiate HK migration. HK cannot properly migrate when plated in the presence of GF, regardless of the concentration, without an ECM substratum. The role of GF, instead, is to augment ECM-initiated motility and provide directionality. To gain insights into the mechanism of action by ECM and GF, we compared, side-by-side, the roles of three major mitogen-activated protein kinase cascades, extracellular-signal-regulated kinase (ERK)1/2, p38, and c-Jun N-terminal kinase (JNK). Our data show that ERK1/2 is involved in mediating collagen's initiation signal and GF's augmentation signal. p38 is specific for GF's augmentation signal. JNK is uninvolved in HK motility. Constitutively activated p38 and ERK1/2 alone could not initiate HK migration. Co-expression of both constitutively activated p38 and ERK1/2, however, could partially mimic the pro-motility effects of collagen and GF. This study reveals for the first time the specific functions of ECM and GF in cell motility.


Characterization of the large deletion in the GALC gene found in patients with Krabbe disease.
Globoid cell leukodystrophy (GLD) of Krabbe disease results from mutations in the galactocerebrosidase (GALC) gene. Previously, we had identified a large deletion in the GALC gene together with a C to T polymorphism at cDNA position 502 in a significant number of cases of infantile Krabbe disease; however, the deletion breakpoint had not been found. In this paper we show that the deletion is approximately 30 kb starting near the middle of the 12 kb intron 10, and includes all of the coding region through exon 17 plus an additional 9 kb. The deletion junction contains a 4 bp direct repeat and is preceded by sequence identified as belonging to the Alu family of interspersed repetitive elements. Using genomic DNA and a PCR-based test to detect normal and deleted sequences at that location, a large number of patients with all clinical types of GLD were analyzed. Of 21 infantile patients found to be heterozygous for the 502T polymorphism reported previously, 15 had the deletion, one could not tested and five, including a Hmong child, did not have the deletion. Sixteen other infantile patients previously tested were found to be either homozygous (10) or heterozygous (6) for the deletion. In addition, five patients with juvenile and adult GLD were found to be heterozygous for the deletion. In every case tested, the deletion was always found on the same allele as the 502T polymorphism. However, other disease-causing mutations have been found on the 502T allele. With careful genotype analysis these families can receive improved genetic information including patient and carrier identification and preimplantation diagnosis.


Neuropathology of autonomic nervous system in Parkinson's disease.
Lewy body formation has been considered to be a marker for neuronal degeneration, because postmortem studies of Parkinson's disease (PD) patients have shown loss of neurons in the predilection sites for Lewy bodies. We systemically studied the autonomic nervous system in patients with PD. Lewy bodies were widely distributed in the hypothalamus, sympathetic system (intermediolateral nucleus of the thoracic cord and sympathetic ganglia) and parasympathetic system (dorsal vagal and sacral parasympathetic nuclei). The number of neurons in the intermediolateral nucleus was significantly reduced. Furthermore, Lewy bodies were also found in the enteric nervous system of the alimentary tract, cardiac plexus, pelvic plexus and adrenal medulla. These findings indicate that both central and peripheral autonomic nervous systems are involved in the disease process in PD.


The importance of airway and lung microbiome in the critically ill.
During critical illness, there are a multitude of forces such as antibiotic use, mechanical ventilation, diet changes and inflammatory responses that could bring the microbiome out of balance. This so-called dysbiosis of the microbiome seems to be involved in immunological responses and may influence outcomes even in individuals who are not as vulnerable as a critically ill ICU population. It is therefore probable that dysbiosis of the microbiome is a consequence of critical illness and may, subsequently, shape an inadequate response to these circumstances.Bronchoscopic studies have revealed that the carina represents the densest site of bacterial DNA along healthy airways, with a tapering density with further bifurcations. This likely reflects the influence of micro-aspiration as the primary route of microbial immigration in healthy adults. Though bacterial DNA density grows extremely sparse at smaller airways, bacterial signal is still consistently detectable in bronchoalveolar lavage fluid, likely reflecting the fact that lavage via a wedged bronchoscope samples an enormous surface area of small airways and alveoli. The dogma of lung sterility also violated numerous observations that long predated culture-independent microbiology.The body's resident microbial consortia (gut and/or respiratory microbiota) affect normal host inflammatory and immune response mechanisms. Disruptions in these host-pathogen interactions have been associated with infection and altered innate immunity.In this narrative review, we will focus on the rationale and current evidence for a pathogenic role of the lung microbiome in the exacerbation of complications of critical illness, such as acute respiratory distress syndrome and ventilator-associated pneumonia.


Further validation of the Self-Defeating Personality Scale.
In a sample of 51 women and 35 men those who scored higher on the Self-defeating Personality Scale reported having "dropped out" of useful activities more often and scored as less assertive on the Assertiveness Self-report Inventory. Women were more likely than men to report having experienced at least one self-defeating relationship. Attempts to replicate gender differences reported earlier were partially successful. The results provide additional validation for the Self-defeating Personality Scale.


Exploring interrelationships between diabetes and periodontal disease in African Americans.
Population-based data on interrelationships between diabetes and periodontal diseases among African Americans are limited. This article is an overview of our knowledge regarding the bidirectional relationship between diabetes and periodontal diseases and a descriptive analysis of data from the Third National Health and Nutrition Examination Survey (NHANES III), focusing on the diabetes-periodontal diseases interrelationship in African Americans. Results of the analysis are consistent with the current body of evidence supporting a bidirectional relationship between diabetes and periodontal diseases and indicate generally poorer periodontal health and glycemic control among African Americans. The results also indicate significantly lower dental care use in dentate African Americans with diabetes than in the US non-Hispanic white population with diabetes.


Effect of leukoreduction and pathogen reduction on the hemostatic function of whole blood.
There is renewed interest in the use of whole blood (WB) for resuscitation of patients in hemorrhagic shock. Leukoreduction with platelet-sparing filters and pathogen reduction may be used to improve the safety profile of WB, yet the effects of leukoreduction and pathogen reduction on WB hemostatic function are not well characterized. Blood from 32 healthy group O donors was divided into treatment groups (n = 8 for each group): untreated, pathogen reduced (PR+ ), leukoreduced using an in-line filter (LR+ ), or PR+ LR+ . Units were stored without agitation for 21 days between 1° and 6°C, with sampling on days 0 (pre- and post-treatments), 1, 3, 5, 10, 15, and 21 for hemostatic function as assessed by thromboelastometry, thrombin generation, platelet activation factors, and platelet impedance aggregometry. From day 3 (D3) to D15 of storage, platelet count was reduced in PR+ /LR+ units compared to PR- /LR- units. From D10 to D21 of storage, maximum clot firmness (MCF) was reduced in PR+ /LR+ units compared to PR- /LR- units. From D3 to D21 of storage, platelet aggregation was reduced in PR+ /LR+ units compared to PR- /LR- units. Total thrombin generation was similar in all groups from D0 to D21. The combination of LR with a platelet-sparing filter and PR significantly reduces hemostatic function compared to either treatment alone or untreated WB. The clinical consequences of LR and PR of WB in patients with severe bleeding should be examined in trials before both are used in combination in patients.


PET Imaging of the Human Nicotinic Cholinergic Pathway in Atherosclerosis.
During the past years, non-neuronal vascular nicotinic acetylcholine receptors (nAChRs) increasingly have gained interest in cardiovascular research, as they are known to mediate the deleterious effects of nicotine and nitrosamines, components of tobacco smoke, on the vasculature. Because smoking is a major risk factor for the development of atherosclerosis, it is obvious that understanding the pathophysiologic role of nAChRs in the atherosclerotic disease process, as well as in the development of new diagnostic and therapeutic nAChR-related options, has become more important. Accordingly, we briefly summarize the pathophysiologic role of vascular nAChRs in the atherosclerotic disease process. We also provide an overview of currently available nAChR positron emission tomography (PET) tracers and their performance in the noninvasive imaging of vascular nAChRs, as well as potential nAChR PET tracers that might be an option for vascular nAChR PET imaging in the future.


"Weight cycling" and mortality: how do the epidemiologists explain the role of intentional weight loss?
In the past 5 years, four prospective, epidemiologic studies of "weight cycling" found that mortality is higher for persons with unstable body weight than for persons whose body weight is relatively stable [1-3,5]. These findings have generated considerable interest and controversy in both the scientific community and the lay press because of the belief that the weight fluctuations were caused by intentional weight loss. None of these studies, however, collected information that would indicate whether the weight changes were intentional or unintentional. This review examines the reasons given by the authors of these studies to support the inference that intentional weight loss caused the increase in mortality. The authors acknowledged that weight loss can be caused by preexisting illness, and they usually made efforts to control for the confounding effects of illness in their analyses. However, they generally concluded that the weight fluctuations resulted from unsuccessful dieting, but no data were presented on the incidence or causes of weight fluctuation in the populations from which the samples were drawn. In none of the studies did the authors recommend that obese persons stop trying to lose weight. The reader is cautioned to await further studies in which intentionality of weight loss is directly assessed before concluding that dieting to reduce weight increases the risk of mortality.


[Role of sympathoadrenal activations in congestive heart failure].
Although sympathoneuronal activation plays a major role in maintaining circulatory homeostasis in decompensated heart failure, protracted activation adversely affects survival in patients with chronic heart failure. Endstage heart failure is associated with beta-adrenergic receptor subsensitivity phenomenon including downregulation of beta1-adrenergic receptors, uncoupling induced by receptor kinase activations, increased level of inhibitory guanine nucleotide binding protein. Abnormal calcium handling occurs as a result of dissociation of ryanodine receptors and its modulatory protein, FK506 binding proteins in addition to defective calcium uptake. Phosphoinositide 3-kinase gamma as well as defective neuronal uptake mechanism of norepinephrine have a pivotal role in mediating these alterations in adrenergic receptor signalings. In this regards, modulation of excessive adrenergic activations is essential to achieve successful management of patients with congestive heart failure.


Resveratrol protects BV2 mouse microglial cells against LPS-induced inflammatory injury by altering the miR-146a-5p/TRAF6/NF-κB axis.
Objective: To investigate the role of miR-146a-5p in the effects of resveratrol (RSV) on inflammatory response in BV2 mouse microglial cells. Materials and methods: BV2 cells were pretreated by RSV and stimulated with lipopolysaccharide (LPS). Cell Viability was checked using a MTT assay. Real-Time PCR was performed to detect the levels of pro-inflammatory cytokines (tumor necrosisfactor-α-TNF-α, interleukin-1β-IL-1β and interleukin-6 - IL-6) and miR-146a-5p expression. Western blot was used to analyze the protein expression of TNF receptor associated factor 6 (TRAF6) and phospho-nuclear factor kappa B (pNF-κB). Gain-of-function and loss-of-function analysis of miR-146a-5p was performed using transfection of miR-146a-5p mimic and miR-146a-5p inhibitor, respectively. Results: Pretreatment with RSV significantly and dose dependently inhibited LPS-induced production of TNF-α, IL-1β and IL-6 in BV2 cells. MiR-146a-5p was significantly upregulated after LPS treatment, and further increased in RSV and LPS-co-treated cells. MiR-146a-5p overexpression via miR-146a-5p mimic transfection downregulated the mRNA level of TNF-α, IL-1β and IL-6, as well as abrogated the protein expression of TRAF6 and pNF-κB in BV2 cells exposed to LPS. More importantly, the reducion of TNF-α, IL-1β and IL-6 level by RSV were reversed by miR-146a-5p silence via miR-146a-5p inhibitor transfection. Furthermore, silencing miR-146a-5p attenuated the inhibitory effect of RSV on the TRAF6/NF-κB pathway which was activated after induction with LPS. Conclusions: RSV can suppress LPS-induced inflammatory injury via modulating the miR-146a-5p/TRAF6/NF-κB axis in BV2 mouse microglial cells.


Breathing patterns in asthmatic children during attack.
Breathing patterns in asthmatic children during attack were studied using multielectrode impedance pneumography. A great number of respiration patterns were analyzed by a personal computer. During severe attacks, a pattern of prolonged expiratory time was noted. Inspiratory time, however, did not change, regardless of the severity of the asthmatic attack. No remarkable changes in breathing patterns were recognized during mild attacks. It was, therefore, considered that breathing patterns remained essentially unchanged except in cases of severe asthmatic attacks.


Identification of PP1alpha as a caspase-9 regulator in IL-2 deprivation-induced apoptosis.
One of the mechanisms that regulate cell death is the reversible phosphorylation of proteins. ERK/MAPK phosphorylates caspase-9 at Thr(125), and this phosphorylation is crucial for caspase-9 inhibition. Until now, the phosphatase responsible for Thr(125) dephosphorylation has not been described. Here, we demonstrate that in IL-2-proliferating cells, phosphorylated serine/threonine phosphatase type 1alpha (PP1alpha) associates with phosphorylated caspase-9. IL-2 deprivation induces PP1alpha dephosphorylation, which leads to its activation and, as a consequence, dephosphorylation and activation of caspase-9 and subsequent dissociation of both molecules. In cell-free systems supplemented with ATP caspase-9 activation is induced by addition of cytochrome c and we show that in this process PP1alpha is indispensable for triggering caspase-9 as well as caspase-3 cleavage and activation. Moreover, PP1alpha associates with caspase-9 in vitro and in vivo, suggesting that it is the phosphatase responsible for caspase-9 dephosphorylation and activation. Finally, we describe two novel phosphatase-binding sites different from the previously described PP1alpha consensus motifs, and we demonstrate that these novel sites mediate the interaction of PP1alpha with caspase-9.


Catalyst and doping methods for arc graphene.
Nitrogen-doped graphene synthesis with ∼g scale has been accomplished using the arc discharge method. The defects formed in the synthesis process were reduced by adding various metal catalysts, among which Bi2O3 was found to be the most effective. Adding dopants to the starting materials increased the electrical conductivity of the graphene product, and the doping concentration in graphene was tuned by adjusting the amount of nitrogen dopants. A step-wise technique to fabricate graphene thin films was developed, including dispersion, separation, and filtering processes. The arc graphene can also find its potential application in supercapacitors, taking advantage of its large surface area and improved conductivity by doping.


Predictors of mortality in chronic pulmonary aspergillosis.
Chronic pulmonary aspergillosis (CPA) is a chronic progressive infection that destroys lung tissue in non-immunocompromised patients. Contemporary series suggest 50-85% 5-year mortality, with few prognostic factors identified.A cohort of 387 CPA patients referred to the UK's National Aspergillosis Centre from 1992 to June 2012 was studied until June 2015. The impact of objective and subjective variables including age, sex, previous pulmonary conditions, dyspnoea score, quality of life, serum albumin and C-reactive protein and radiological appearances were assessed using Kaplan-Meier curves, log rank tests and Cox proportional hazards modelling. In samples of patients, retrospective review of time from likely onset of CPA to referral and cause of death were also investigated.Survival was 86%, 62% and 47% at 1, 5 and 10 years, respectively. Increased mortality was associated with nontuberculous mycobacterial infection (hazard ratio 2.07, 95% CI 1.22-3.52; p<0.001) and chronic obstructive pulmonary disease (1.57, 1.05-2.36; p=0.029) as well as higher age (1.053, 1.03-1.07 per year; p<0.001), lower albumin (0.92, 0.87-0.96 per g·L-1), lower activity (1.021, 1.01-1.03 per point increase in St George's Respiratory Questionnaire activity domain; p<0.001) and having one, and especially, bilateral aspergillomas (p<0.001).Several factors impact on mortality of CPA, and can be evaluated as tools to assess CPA prognosis.


CGG repeat RNA G-quadruplexes interact with FMRpolyG to cause neuronal dysfunction in fragile X-related tremor/ataxia syndrome.
Fragile X-related tremor/ataxia syndrome (FXTAS) is a neurodegenerative disease caused by CGG triplet repeat expansions in FMR1, which elicit repeat-associated non-AUG (RAN) translation and produce the toxic protein FMRpolyG. We show that FMRpolyG interacts with pathogenic CGG repeat-derived RNA G-quadruplexes (CGG-G4RNA), propagates cell to cell, and induces neuronal dysfunction. The FMRpolyG polyglycine domain has a prion-like property, preferentially binding to CGG-G4RNA. Treatment with 5-aminolevulinic acid, which is metabolized to protoporphyrin IX, inhibited RAN translation of FMRpolyG and CGG-G4RNA-induced FMRpolyG aggregation, ameliorating aberrant synaptic plasticity and behavior in FXTAS model mice. Thus, we present a novel therapeutic strategy to target G4RNA prionoids.


[Effect of electroacupuncture on expression of stromal cell derived factor-1α in focal ischemic cerebral tissue of rats with cerebral ischemia/ reperfusion injury].
To observe the effect of electroacupuncture (EA) of "Baihui" (GV20) and "Zusanli" (ST36) on the expression of stromal cell derived factor-1α (SDF-1α) and CD34 in ischemic cortex tissue of cerebral ische-mia /reperfusion injury (CI/RI) rats, so as to study its mechanisms underlying improving CI/RI. Thirty male SD rats were equally and randomly divided into sham operation, model and EA groups (n=10 rats in each group). The CI/RI model was established by occlusion of the middle cerebral artery for 120 min, followed by reperfusion. EA (40 Hz, 1-2 mA) was applied to GV20 and left ST36 for 20 min, once daily for successive 14 days. The neurological deficit severity was assessed by using Longa's and colleagues' methods. The histopathological changes of the ischemic tissues were observed after H.E. staining and the expression of SDF-1α and CD34 in the ischemic cortex tissues was detected by immunohistochemistry. After modeling, the neurological deficit score, and the numbers of SDF-1α and CD34-positive cells in the ischemic cerebral cortex tissue were significantly increased in the model group (P<0.05). Following EA intervention, the increased neurological deficit score was reversed at the 3rd, 7th and 14th day, and the increased SDF-1α and CD34-positive cells were significantly further up-regulated in the EA group (P<0.05). H.E. staining showed tissue edema, widening of the intercellular space, cavitation-like changes, neuronal shrinkage, nuclear pyknosis with hyperchroma or even disappearance, and aggregation of inflammatory and neurogliocytes in the model group. These situations were relatively milder in the EA group. EA of GV20 and ST36 can improve neurological function of CI/RI rats, which may be associated with its effect in up-regulating the expression of SDF-1α and CD34 proteins in the ischemic cerebral cortex tissues.


Long non-coding RNA H19 correlates with unfavorable prognosis and promotes cell migration and invasion in ovarian cancer.
The purpose of this study is to investigate the expression pattern of lncRNA H19 in OC tissues and to detect the ability of H19 to influence OC cell migration and invasion in vitro. We quantified the levels of H19 within the obtained cancerous and adjacent noncancerous tissues from 258 OC patients. H19 association with patient progression-free survival (PFS) was analyzed by a Kaplan-Meier plot. Expression levels of H19 were reduced by small interfering RNA transfection against H19 or restored by a H19 overexpression plasmid transfection in OC cells. H19 effects on OC cell migration and invasion in vitro were evaluated using wound-healing assay and transwell invasion assay. Wound healing assay and transwell invasion assay were used to evaluate the effects of H19 on OC cell migration and invasion in vitro. H19 is upregulated remarkably in primary OC tissues and human OC cell lines (OVCAR3, SKOV3, A2780, and Caov-3). We found that the median PFS was longer in patients with lower levels of H19 than in those with high levels, suggesting that overexpression of H19 was linked to poor prognosis in OC patients. Intriguingly, the depletion of H19 expression induced by small interfering RNA inhibited the capability of migration and invasion of OC cell lines. Restoration of H19 in OC cell lines significantly increased cell migration and invasion. The key finding of the present study suggests that overexpression of H19 may be associated with an unfavorable prognosis for OC and is likely to be a possible contributory force involved in OC cell migration and invasion. H19 may provide a new and attractive target for future prognostic and therapeutic intervention of OC patients.


Brain Network Organization Following Post-Stroke Neurorehabilitation.
Brain network analysis can offer useful information to guide the rehabilitation of post-stroke patients. We applied functional network connection models based on multiplex-multilayer network analysis (MMN) to explore functional network connectivity changes induced by robot-aided gait training (RAGT) using the Ekso, a wearable exoskeleton, and compared it to conventional overground gait training (COGT) in chronic stroke patients. We extracted the coreness of individual nodes at multiple locations in the brain from EEG recordings obtained before and after gait training in a resting state. We found that patients provided with RAGT achieved a greater motor function recovery than those receiving COGT. This difference in clinical outcome was paralleled by greater changes in connectivity patterns among different brain areas central to motor programming and execution, as well as a recruitment of other areas beyond the sensorimotor cortices and at multiple frequency ranges, contemporarily. The magnitude of these changes correlated with motor function recovery chances. Our data suggest that the use of RAGT as an add-on treatment to COGT may provide post-stroke patients with a greater modification of the functional brain network impairment following a stroke. This might have potential clinical implications if confirmed in large clinical trials.


Preliminary assessment and treatment guidelines for hip disorders in young adults.
The treatment of hip disorders in adolescents and young adults has evolved rapidly during the past decade because of enhanced understanding of prearthritic and early arthritic hip disease combined with improved diagnostic skills, better patient selection criteria, and refinements in alternative surgical procedures. Despite these advancements, the diverse clinical conditions and available surgical options highlight the need to develop optimal diagnostic and treatment guidelines for young patients with compromised hip function. Comprehensive care of young patients with hip disorders includes diverse surgical techniques such as arthroscopy, pelvic osteotomies, osteoplasty, femoral osteotomies, surgical dislocation, grafting procedures, arthrodesis, and prosthetic replacement. Optimal clinical results depend on the combination of careful patient selection and successful application of the appropriate surgical procedure. We outline general guidelines for the assessment and treatment of hip pain in young adult patients.


Tumor penetrating nanomedicine targeting both an oncomiR and an oncogene in pancreatic cancer.
Developing new targeted therapy for pancreatic cancer is one of the major current challenges in cancer research. KRAS mutations and miRNA dysregulation (e.g. miR-21-5p oncomiR) play key roles in Pancreatic Ductal Adenocarcinoma (PDAC), leading to rapid progression of the disease. As the KRAS mutation is a main driver of PDAC, anti-KRAS strategies remain a major therapeutic approach for PDAC treatment. Previously, utilization of either siKRAS or small chemically modified single-stranded RNA molecules that specifically disable miR-21 (anti-miR-21) were effective in slowing PDAC tumor growth in various tumor models when packaged in an innovative delivery system (TPN) required for efficient drug delivery to the PDAC tumor site. Here we have tested the utility of targeting the KRAS pathway through multiple mechanisms and via dual targeting of a PDAC oncomiR and oncogene. Initially we found that miR-217, which has been shown to directly regulate KRAS expression, is downregulated in our PDAC samples, thus we tested the benefits of anti-miR-21, miR-217 mimic or siKRAS loaded into the tumor-penetrating nanoparticles (TPN) that we had previously shown to potently target the largely impenetrable PDAC tumors, and found an enhanced anti-tumoral response upon dual treatments in KRAS-mutated PDAC models.


Increase of condylar displacement between centric relation and maximal habitual intercuspation after occlusal splint therapy.
The present study assessed condylar displacement between initial maximal habitual intercuspation (MHI) and centric relation (CR), recorded after using a deprogramming occlusal splint for an average period of 7.8 +/- 2.1 months prior to any orthodontic treatment. The sample consisted of 22 subjects, 11 male and 11 female, with an average age of 14.2 +/- 1.4 years, with Class II malocclusion and with no apparent signs or symptoms of temporomandibular dysfunction (TMD). Condylar displacement was measured using a Panadent axis position indicator in decimal fractions of a millimeter. The original mean vertical displacements and the corresponding standard deviations were 4.24 +/- 2.53 mm and 3.86 +/- 2.72 mm, respectively, for the right and left sides. Because a significant negative correlation was observed between original condylar displacements and age factors, the displacement values were statistically adjusted to 2.74 +/- 2.00 mm and 2.44 +/- 1.93 mm. On the horizontal plane, the mean displacements measured were -0.72 +/- 1.53 mm on the right side and -0.51 +/- 1.98 mm on the left. The mean displacement on the transversal plane was 0.03 +/- 0.87 mm. A comparison between these values and those observed in non-deprogrammed groups, as well as those published in the related literature, indicates that use of occlusal splints results in greater mean condylar displacement values, especially vertically, between CR and MHI positions, which contributed to a more accurate orthodontic diagnosis.


Extension of the Kubelka-Munk theory to an arbitrary substrate: a Monte Carlo approach.
In this work we review and-to some extent-upgrade one of the main theories of light flux through homogeneous isotropic media, namely, the Kubelka-Munk (K-M) theory, and in particular the later expansion made by Kubelka to obtain the reflectance of a specimen when a substrate lies underneath. We have completed this solution by calculating the transverse energy density in the specimen and the transmission of the whole. We show that this last result-compatible with Kubelka's upgrade for layered media-also allows for the calculation of the specimen/substrate absorption split. In order to validate these expressions, the results were reproduced by means of a Monte Carlo simulation working on a layered medium under the same assumptions as the K-M theory. Interestingly, the numerical procedure introduces new capabilities in the model regarding the history of any absorbed or outgoing elemental light beam, such as the recording of its time-of-flight through a given system.


Transgenic switchgrass (Panicum virgatum L.) biomass is increased by overexpression of switchgrass sucrose synthase (PvSUS1).
Sucrose synthase (SUS) converts sucrose and uridine di-phosphate (UDP) into UDP-glucose and fructose. UDP-glucose is used by the cellulose synthase to produce cellulose for cell wall biosynthesis. For lignocellulosic feedstocks such as switchgrass, the manipulation of cell walls to decrease lignin content is needed to reduce recalcitrance of conversion of biomass into biofuels. Of perhaps equal importance for bioenergy feedstocks is increasing biomass. Four SUS genes were identified in switchgrass. Each gene contained 14 or 15 introns. PvSUS1 was expressed ubiquitously in the tissues tested. PvSUS2 and PvSUS6 were highly expressed in internodes and roots, respectively. PvSUS4 was expressed in low levels in the tissues tested. Transgenic switchgrass plants overexpressing PvSUS1 had increases in plant height by up to 37%, biomass by up to 13.6%, and tiller number by up to 79% compared to control plants. The lignin content was increased in all lines, while the sugar release efficiency was decreased in PvSUS1-overexpressing transgenic switchgrass plants. For switchgrass and other bioenergy feedstocks, the overexpression of SUS1 genes might be a feasible strategy to increase both plant biomass and cellulose content, and to stack with other genes to increase biofuel production per land area cultivated.


Using pens as an incentive for trial recruitment of older adults: An embedded randomised controlled trial.
Background: Meeting recruitment targets for randomised controlled trials is challenging. This trial evaluated the effectiveness of including a pen within the trial invitation pack on the recruitment of older adults into a randomised controlled trial. Methods: This trial was embedded within the Occupational Therapist Intervention Study, a falls-prevention randomised controlled trial. Potential participants (n = 1862), who were posted an invitation pack from two General Practitioner practices, were randomised to either not receive a pen (n = 1295) or receive a pen (n = 648) with their invitation pack, using a 2:1 ratio. The primary outcome was the likelihood of being randomised, and therefore fully recruited, to the host trial. To be randomised to the host trial, participants had to: return a consent form and screening form; be eligible on their screening form; and return a baseline questionnaire and a monthly falls calendar. Secondary outcomes were: the likelihood of returning (and time to return) a screening form; being eligible for the host trial; and remaining in the trial for at least 3 months. Results: The likelihood of being randomised to the host trial did not differ between the pen group (4.5%) and no pen group (4.3%; odds ratio 1.04; 95% confidence interval: 0.65 to 1.67; p = 0.86). There were marginal differences in secondary outcomes in favour of the pen group, particularly in screening form return rates, though these differences were not statistically significant. Conclusion: Pens may not be an effective incentive for the recruitment of older adults into randomised controlled trials, though future trials are required. Registration: ISRCTN22202133; SWAT 37.


Hepatitis B, aflatoxin B(1), and p53 codon 249 mutation in hepatocellular carcinomas from Guangxi, People's Republic of China, and a meta-analysis of existing studies.
The incidence of hepatocellular carcinomas (HCC) varies widely worldwide, with some of the highest incidence rates found in China. Chronic infection with the hepatitis B virus (HBV) and exposure to aflatoxins in foodstuffs are the main risk factors. A G to T transversion at codon 249 of the p53 gene (249(ser)) is commonly found in HCCs from patients in regions with dietary aflatoxin exposure. Because HBV infection is often endemic in high aflatoxin exposure areas, it is still unclear whether HBV acts as a confounder or as a synergistic partner in the development of the 249(ser) p53 mutation. Our report has two aims. First, we contribute data on HCCs from southern Guangxi, a high aflatoxin exposure area. Using DNA sequencing, we found that 36% (18 of 50) of tumors had a 249(ser) mutation. Also, 50% (30 of 60) were positive for p53 protein accumulation and 78% (28 of 36) were positive for HBV surface antigen, as detected by immunohistochemistry. Second, we present a meta-analysis, using our results along with those from 48 published studies, that examines the interrelationships among aflatoxin exposure, HBV infection, and p53 mutations in HCCs. We used a method that takes into account both within-study and study-to-study variability and found that the mean proportion of HCCs with the 249(ser) mutation was positively correlated with aflatoxin exposure (P = 0.0001). We found little evidence for an HBV-aflatoxin interaction modulating the presence of the p53 249(ser) mutation or any type of p53 mutation.


Revolutions in ophthalmology.
Cataract is the major cause of blindness worldwide. The transition of couching of cataract to most modem phaco-emulsification gained popularity with more scientific capsulorhexis. Ongoing research is taking place with laser technologies for removal of cataract. Surgery has an important role in advanced corneal diseases. Corneal transplantation is the most successful form. The refractive errors can be corrected by laser-assisted in-situ keratomileusis, intrastromal corneal rings and phacic intra-ocular lenses. Current research is going on to identify the gene responsible for glaucoma and once this is identified, medical therapy can be developed to control the disease. Thermocautery or photocoagulation and indenting the globe so as to bring the choroid nearer the break by scleral buckling forms the basis of modern surgery of retinal detachment. Vitreoretinal diseases, macular degeneration, diabetic retinopathy are no longer considered problematic. We are optimistic that more clinical breakthroughs are expected in the 21st century with new advances of ophthalmic researches.


Linked production of antibodies to mammalian DNA and to human polyomavirus large T antigen: footprints of a common molecular and cellular process?
To test whether the presence of antibodies to human polyomavirus large T antigen, a viral DNA-binding protein essential for productive polyomavirus replication, correlates with the presence of antibodies to single-stranded DNA (ssDNA), double-stranded DNA (dsDNA), or the autologous TATA-binding protein (TBP). Sera from patients with various diagnosed or suspected autoimmune syndromes were analyzed for the presence of antibodies to T antigen, DNA, or TATA-binding protein, and correlations were determined. Rheumatoid factor (RF) was studied as a control antibody. A highly significant correlation between antibodies to T antigen and antibodies to ssDNA or TATA-binding protein, but not between anti-T antigen antibodies and RF, was found in all patient groups. Of all sera that were positive for antibodies to dsDNA, 62% were positive for antibodies to T antigen (P<0.03). A non-self DNA-binding protein such as human polyomavirus large T antigen may render DNA immunogenic upon binding to nucleosomes when expressed in vivo. This is indicated by the strong correlation between antibodies to T antigen and antibodies to DNA or TBP and is consistent with a hapten-carrier model. This model implies cognate antigen-selective interaction of T antigen-specific T helper cells and DNA-specific B cells or B cells specific for other components of nucleosomes, consistent with the results of previous experiments.


Purification of xanthine dehydrogenase from rat liver: a rapid procedure with high enzyme yields.
Xanthine dehydrogenase (EC 188.8.131.52) was purified approximately 1000-fold from liver homogenates of adult male Sprague-Dawley rats. Enzyme recovery was good (greater than 20% of the starting activity was obtained), and the homogeneously pure enzyme had a molecular mass of approximately 300,000 Da. The purified protein exhibited a specific activity of 2470 units/mg protein and spectral properties identical to those of the best preparations of this enzyme reported by other investigators. Routine preparations of this enzyme also possess higher dehydrogenase:oxidase ratios (typically between 5 and 6) than do other xanthine dehydrogenase preparations so far reported in the literature. Maximum dehydrogenase:oxidase ratios, greater than 10, could be obtained from this procedure if only peak dehydrogenase fractions from the chromatography columns were saved. The present small-scale purification method, which can be completed in 48-60 h, utilizes ammonium sulfate fractionation, Sephadex G-200 column chromatography, Blue Dextran-Sepharose column chromatography, and preparative gel electrophoresis.


Factors influencing willingness to pay for accident risk reduction among personal car drivers in Thailand.
Thailand ranks near the top for the road accident fatality rate worldwide, and more and more vehicles are being registered in Thailand every year. Obtaining the opinions of road commuters may help us reduce road accidents in Thailand. This study seeks to understand damage value in road accidents for personal car drivers in Thailand, using the willingness to pay approach and establishing factors affecting willingness to pay with the theory of planned behavior (TPB). This study obtained data using questionnaires in face-to-face interviews with 1,650 personal cars drivers in Thailand. The average willingness to pay (WTP) for 50% fatality or injury reduction was 23.00 baht/person/50 km trip (US $0.74/person/50 km trip). We obtained the value of statistical life (VSL), assessing this to fall between US $815,385 and US $872,942, and the value of statistical injury (VSI), between US $150,059 and US $160,652. Overall, national damage was assessed at US $4,701,981,170 annually. According to the analysis of factors affecting WTP, TPB comprises four factors, namely, driver attitude, subjective norm, perceived behavioral control, and behavioral intention. Analysis using structural equation modeling (SEM) found all mentioned factors were relevant and positively influenced personal car drivers' WTP in Thailand, with a statistical significance at a 99% confidence interval (p < 0.01). This study can develop recommendations for relevant organizations to analyze the results as part of considerations regarding budget allocation and developments on road safety policy due to driver attitude as important as environmental factors or any other factors.


[Study of adiponectin expression in placenta and its correlation with preeclampsia].
To investigate the expression of adiponectin in placenta and its correlation with preeclampsia. Placental tissues were collected from normal term pregnancies (normal pregnancy group, n = 20), mild preeclampsia (mild preeclampsia group, n = 12) and severe preeclampsia (severe preeclampsia group, n = 22). The expression of adiponectin protein and the intensity of its mRNA in placenta were detected using immunohistochemistry and RT-PCR, respectively. Integral optical density (IOD) which represents the expression level of adiponectin protein, and the ratio of adiponectin cDNA PCR products to beta-actin cDNA PCR products which represents the intensity of transcription of adiponectin mRNA in placenta were analyzed. (1) The expression of adiponectin protein was observed in cytoplasm of placental cytotrophoblasts and syncytiotrophoblasts among three groups. There was no significant difference in adiponectin protein expression between maternal side and fetal side of placenta in three groups (all P > 0.05); (2) The expression of adiponectin protein in placenta in severe preeclampsia group (30 984 +/- 14 604) was significantly lower than that of mild preeclampsia group (58 360 +/- 8910, P < 0.01) and of normal pregnancy group (53 246 +/- 17 554, P < 0.01). There was also no significant difference in the expression of adiponectin protein in placenta between term delivery and preterm delivery in severe preeclampsia group (38 890 +/- 20 386 vs 29 319 +/- 8997, P > 0.05), however, the expression of adiponectin protein in placenta in term delivery of severe preeclampsia group was significantly lower than that of term delivery of normal pregnancy group (38 890 +/- 20 386 vs 53 246 +/- 17 554, P < 0.05); (3) The expression of adiponectin mRNA was detected in placental tissues among three groups also. The intensity of transcription of adiponectin in placenta in severe preeclampsia group (1.0 +/- 0.2) was markedly lower than that of mild preeclampsia group (2.9 +/- 0. 8, P < 0.05) and normal pregnancy group (3.3 +/- 1.1, P = 0.000). The expression of adiponectin decreases in placenta tissues of severe preeclampsia, indicating that the abnormal expression of adiponectin may be involved in the pathogenesis of preeclampsia.


Interactions of the Hindgut Mucosa-Associated Microbiome with Its Host Regulate Shedding of Escherichia coli O157:H7 by Cattle.
Cattle are the primary carrier of Escherichia coli O157:H7, a foodborne human pathogen, and those shedding >104 CFU/gram of feces of E. coli O157:H7 are defined as supershedders (SS). This study investigated the rectoanal junction (RAJ) mucosa-associated microbiota and its relationship with host gene expression in SS and in cattle from which E. coli O157:H7 was not detected (nonshedders [NS]), aiming to elucidate the mechanisms involved in supershedding. In total, 14 phyla, 66 families, and 101 genera of RAJ mucosa-associated bacteria were identified and Firmicutes (61.5 ± 7.5%), Bacteroidetes (27.9 ± 6.4%), and Proteobacteria (5.5 ± 2.1%) were the predominant phyla. Differential abundance analysis of operational taxonomic units (OTUs) identified 2 OTUs unique to SS which were members of Bacteroides and Clostridium and 7 OTUs unique to NS which were members of Coprococcus, Prevotella, Clostridium, and Paludibacter Differential abundance analysis of predicted microbial functions (using PICRUSt [phylogenetic investigation of communities by reconstruction of unobserved states]) revealed that 3 pathways had higher abundance (log2 fold change, 0.10 to 0.23) whereas 12 pathways had lower abundance (log2 fold change, -0.36 to -0.20) in SS. In addition, we identified significant correlations between expression of 19 differentially expressed genes and the relative abundance of predicted microbial functions, including nucleic acid polymerization and carbohydrate and amino acid metabolism. Our findings suggest that differences in RAJ microbiota at both the compositional and functional levels may be associated with E. coli O157:H7 supershedding and that certain microbial groups and microbial functions may influence RAJ physiology of SS by affecting host gene expression.IMPORTANCE Cattle with fecal E. coli O157:H7 at >104 CFU per gram of feces have been defined as the supershedders, and they are responsible for the most of the E. coli O157:H7 spread into farm environment. Currently, no method is available for beef producers to eliminate shedding of E. coli O157:H7 in cattle, and the lack of information about the mechanisms of supershedding greatly impedes the development of effective methods. This study investigated the role of the rectoanal junction (RAJ) mucosa-associated microbiome in E. coli O157:H7 shedding, and our results indicated that the compositions and functions of RAJ microbiota differed between supershedders and nonshedders. The identified relationship between the differentially abundant microbes and 19 previously identified differentially expressed genes suggests the role of host-microbial interactions involved in E. coli O157:H7 supershedding. Our findings provide a fundamental understanding of the supershedding phenomenon which is essential for the development of strategies, such as the use of directly fed microbials, to reduce E. coli O157:H7 shedding in cattle.


Case of megaloblastic anemia caused by intestinal taeniasis.
A 61-year-old eutrophic male was diagnosed with vitamin B12 deficiency and megaloblastic anemia. A modified Schilling test suggested intestinal malabsorption unrelated to intrinsic factor deficiency. Subsequent colonoscopy revealed the presence of a Taenia tapeworm. The anemia resolved within days under therapy with niclosamide and temporary vitamin B12 supplements. The present case suggests that, in addition to other well-known parasitic agents, e.g., Diphyllobothrium latum and Giardia lamblia, Taenia infestation can also be a cause of intestinal vitamin B12 malabsorption.


Influenza vaccination induced leukocytoclastic vasculitis and pauci-immune crescentic glomerulonephritis.
Influenza vaccination is a widely accepted practice, particularly among the elderly and high-risk individuals. Minor and transitory side effects following the vaccination are common, while systemic complications are infrequently reported. We describe here a case of a patient who presented to the emergency room with arthralgia, myalgias and purpura, following influenza vaccination. Necrotizing vasculitis associated with pauci-immune glomerulonephritis was observed on kidney biopsy. With increasing use of influenza vaccination, attention should be drawn to the possible expression of systemic adverse effects such as vasculitis and glomerulonephritis.


Mechanism of bacterial bioluminescence: 4a,5-dihydroflavin analogs as models for luciferase hydroperoxide intermediates and the effect of substituents at the 8-position of flavin on luciferase kinetics.
Bioluminescence catalyzed by bacterial luciferases was measured using FMN, iso-FMN (6-methyl-8-nor-FMN), and FMN analogs carrying the following substituents at position 8: -H, -Cl, -F, SMe, SOMe, -SO2Me, or -OMe. The first-order rate constants for the decay of light emission correlate with the one-electron oxidation potentials of the 4a,5-dihydro forms of the FMN analogs. To determine the values of these potentials, isoalloxazine (flavin) derivatives having the 4a,5-propano-4a,5-dihydro structure and -H, -CH3, -Cl, -OCH3, and -NH2 as substituents at position 8 have been synthesized as models for the 4a-peroxy-4a,5-dihydroflavin intermediates occurring during catalysis by the flavin-dependent monooxygenase luciferase. The tetrahydropyrrole ring between positions 4a and 5 of these isoalloxazine derivatives stabilizes the 4a,5-dihydroflavin by impeding formation of the thermodynamically more stable 1,5-dihydro form. One-electron oxidation potentials (Eobs) were measured by cyclic voltammetry and used to determine the empirical coefficients in the Swain equation. On the basis of this, the one-electron oxidation potentials of 4a,5-propano-4a,5-dihydro analogs with other substituents in position 8 were calculated (Ecalc). The bioluminescence reaction rate is fastest with FMN analogs of lowest oxidation potential; i.e., the slope of the correlation is negative. This indicates that in the rate-limiting step the 4a,5-dihydroflavin moiety donates negative charge. The results are compatible with an intramolecular, chemically initiated electron exchange luminescence mechanism for the bacterial luciferase reaction.(ABSTRACT TRUNCATED AT 250 WORDS)


Shift in diversification in sister-clade comparisons: a more powerful test.
Tests of shift in diversification associated with key innovations or directional environmental change can be performed with sister-clade comparisons. This approach is attractive because it does not require detailed phylogenetic information. I propose a new likelihood ratio test based on fitting two models of diversification. I show how this test differs from a previous likelihood ratio test based on the geometric distribution. With simulations from a wide range of situations, I show that the new test performs much better than this test and the traditional test by Slowinski and Guyer. The proposed test performs at least as well as the species richness contrast test that has been proposed by several authors in four versions. A power analysis with low number of pairs of sister clades showed that the new test could detect a shift in diversification with five or less pairs of sister clades, whereas the diversity contrast test cannot detect any shift in this situation. The former appears as more powerful than the latter, and therefore is recommended when the number of pairs of sister clades is low (less than 10). All other tests should not be used as the present study showed they lack statistical power and robustness.


Rhizobium infection: lessons from the versatile nodulation behaviour of water-tolerant legumes.
Water-tolerant legumes provide bacteria with special ways of invading roots to establish N(2)-fixing symbiosis upon flooding. On well-aerated roots, root hair curling (RHC) invasion is used, whereas, under hydroponic conditions, rhizobia enter the cortex through cracks at lateral root bases (LRBs). Here, we compare the physiological and anatomical traits of these invasions. During waterlogging, accumulating ethylene inhibits the epidermal stages of RHC invasion. LRB invasion circumvents this step by direct colonization of the cortical tissue. By avoiding the epidermis for bacterial entry under hydroponic conditions, the stringent nodulation (Nod) factor perception systems that are active within the epidermis are not needed. Consequently, LRB invasion might be useful for analysing the requirement for Nod factor perception and other signal transduction systems downstream of the epidermis.


Third order aberrations in cassegrain-type telescopes and coma correction in servo-stabilized images.
Pointing errors in telescopes carried by space vehicles can be compensated by servo-control of the secondary mirror. This introduces some aberrations in the stabilized image, the magnitude of which depends on the asphericity of the telescope mirrors. A configuration is presented in which the centered image of an off-axis object point is free from coma. The required aspheric corrections and the residual aberrations are calculated. Also included is a summary of the third order aberration constants for some well-known centered cassegrain-type telescopes.


Conformation-specific pathways of beta-alanine: a vacuum ultraviolet photoionization and theoretical study.
We report a photoionization and dissociative photoionization study of beta-alanine using IR laser desorption combined with synchrotron vacuum ultraviolet (VUV) photoionization mass spectrometry. Fragments at m/z = 45, 44, 43, and 30 yielded from photoionization are assigned to NH(3)CH(2)CH(2)(+), NH(2)CHCH(3)(+), NH(2)CHCH(2)(+), and NH(2)CH(2)(+), respectively. Some new conformation-specific dissociation channels and corresponding dissociation energies for the observed fragments are established and determined with the help of ab initio G3B3 calculations and measurements of photoionization efficiency (PIE) spectra. The theoretical values are in fair agreement with the experimental results. Three low-lying conformers of the beta-alanine cation, including two gauche conformers G1+, G2+ and one anti conformer A+ are investigated by G3B3 calculations. The conformer G1+ (intramolecular hydrogen bonding N-H...OC) is found to be another precursor in forming the NH(3)CH(2)CH(2)(+) ion, which is complementary to the previously reported formation pathway that only occurs with the conformer G2+ (intramolecular hydrogen bonding O-H...N). Species NH(2)CHCH(2)(+) may come from the contributions of G1+, G2+, and A+ via different dissociation pathways. The most abundant fragment ion, NH(2)CH(2)(+), is formed from a direct C-C bond cleavage. Intramolecular hydrogen transfer processes dominate most of the fragmentation pathways of the beta-alanine cation.


In situ anticariogenic potential of glass ionomer cement.
The purpose of this study was to compare the amount of fluoride in plaque formed on glass ionomer cement or composite and to evaluate the effects of fluoride released on growth of cariogenic microflora, fluoride uptake, and secondary caries formation under in situ conditions of a high cariogenic challenge. Ten adult volunteers took part in this crossover study performed in two phases of 28 days. Eighty enamel blocks were randomly restored with glass ionomer cement (Chelon-Fil-Espe) or composite (Silux). During each phase of the study, an acrylic resin appliance, containing four enamel blocks restored with the same material, was constructed for each of the volunteers. During the experimental period, all subjects used fluoride-free dentifrice, refrained from brushing the restored enamel blocks, and immersed the appliances into 20% sucrose solution eight times a day. Fluoride levels, mutans streptococci, and lactobacilli were assessed in dental plaque. Fluoride uptake and microhardness profiles were determined in enamel around the restorations. Statistical analyses indicated a significantly higher level of fluoride (p < 0.05) and a lower level of mutans streptococci plaque formed on glass ionomer cement. Analysis of variance in a split-plot model indicated that in the enamel around the glass ionomer restoration the fluoride uptake was significantly greater (p < 0.025) and the mineral loss significantly lower (p < 0.01). The results show that glass ionomer cement presents a broad anticariogenic effect and may be of value in preventing secondary caries, even under conditions of a high caries risk.


Response to second treatment after initial failed treatment in a multicenter prospective infantile spasms cohort.
Infantile spasms (IS) represent a severe epileptic encephalopathy presenting in the first 2 years of life. Recommended first-line therapies (hormonal therapy or vigabatrin) often fail. We evaluated response to second treatment for IS in children in whom the initial therapy failed to produce both clinical remission and electrographic resolution of hypsarhythmia and whether time to treatment was related to outcome. The National Infantile Spasms Consortium established a multicenter, prospective database enrolling infants with new diagnosis of IS. Children were considered nonresponders to first treatment if there was no clinical remission or persistence of hypsarhythmia. Treatment was evaluated as hormonal therapy (adrenocorticotropic hormone [ACTH] or oral corticosteroids), vigabatrin, or "other." Standard treatments (hormonal and vigabatrin) were compared to all other nonstandard treatments. We compared response rates using chi-square tests and multivariable logistic regression models. One hundred eighteen infants were included from 19 centers. Overall response rate to a second treatment was 37% (n = 44). Children who received standard medications with differing mechanisms for first and second treatment had higher response rates than other sequences (27/49 [55%] vs. 17/69 [25%], p < 0.001). Children receiving first treatment within 4 weeks of IS onset had a higher response rate to second treatment than those initially treated later (36/82 [44%] vs. 8/34 [24%], p = 0.040). Greater than one third of children with IS will respond to a second medication. Choosing a standard medication (ACTH, oral corticosteroids, or vigabatrin) that has a different mechanism of action appears to be more effective. Rapid initial treatment increases the likelihood of response to the second treatment.


Diphenylphosphinoyl chloride as a chlorinating agent--the selective double activation of 1,2-diols.
Treatment of 1,2-diols with diphenylphosphinoyl chloride in pyridine produces beta-chloroethyl phosphinates which react with complete control of stereochemistry to give epoxides and azido-alcohols, useful intermediates in cyclopropane synthesis.