Behavioral, emotional, and situational context of purging episodes in anorexia nervosa.
The current study examined behavioral, emotional, and situational factors involved in purging among women with anorexia nervosa (AN). Women with AN (n=118) completed a two-week ecological momentary assessment protocol involving daily reports of eating disorder behaviors, mood, and stressful events. Generalized estimating equations examined the likelihood and context of purging following eating episodes involving both overeating and loss of control (binge eating; BE); loss of control only (LOC); overeating only (OE); and neither loss of control nor overeating (non-pathological eating; NE). Relative to NE, purging was more likely to occur following BE, LOC, and OE (Wald chi-square = 18.05; p < .001). BE was more strongly associated with subsequent purging than LOC but not OE; the latter two did not differ from one another. Negative affect predicted purging following NE (Wald chi-square = 7.71; p = .005). Binge eating involving large amounts of food was the strongest predictor of purging in AN, which challenges the notion that loss of control is the most salient aspect of experiencing distress in bulimia nervosa and BE disorder. Parallel to findings from the BE literature, negative affect strongly predicted purging following NE. Further research should clarify the function and triggers of purging in AN.


Preoperative Management of a Jehovah's Witness Patient Undergoing Head and Neck Surgery With High Risk of Bleeding.
Surgery in head and neck district is often associated with bleeding as major complication and need of blood transfusions. Homologous blood transfusions may be unacceptable to some patient groups such as the Jehovah's Witness (JW) patients. Refusal of potentially life-saving treatment creates ethical dilemmas for treating clinicians.This is the first report in literature which examines the management and treatment of a female JW patient who underwent major surgical procedure for squamous cell carcinoma of the jaw with a high risk of hemorrage which rejected any possibility of blood and hemocomponent transfusion by virtue of her religious principles.


Formulation and In Vitro Characterization of Bioactive Mesoporous Silica with Doxorubicin and Metronidazole Intended for Bone Treatment and Regeneration.
The purpose of this study was to evaluate the surface mineralization activity and in vitro drug behavior potential of two forms of mesoporous silica: powder and granulate. Ordered mesoporous SiO2 powder was synthesized by surfactant-assisted sol-gel process using tetraethoxysilane as a silica precursor and hexadecyltrimethylammonium bromide as the structure-directing agent. The granulate was prepared using silica powder and ethyl cellulose as a binding agent. Metronidazole (MT)-an anti-inflammatory substance and doxorubicin hydrochloride (ChD)-an anti-cancer drug were chosen as drug models for delivery studies. The results of structural characteristic studies, utilizing transmission electron microscope (TEM) and scanning electron microscope (SEM) images, powder X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and nitrogen adsorption-desorption (BET) measurements, show that obtained materials have two-dimensional hexagonal p6mm symmetry, high specific surface area, narrow pore size, and a satisfactory mineralization behavior in the simulated body solution (SBF, pH = 7.4). The release rate of drugs depends upon the structural features of the drug molecules and the form of the carrier material. Of both the drugs analyzed, faster release was observed for small MT molecules characterized by weaker interactions with the carrier. In addition, the slower drug release was observed with granulate form due to increased diffusion barrier for drugs. Obtained results prove that the MT/ChD-loaded silica formulations could be attractive materials for filling bone defects and for local delivery systems.


Epigallocatechin-3-O-gallate inhibits fibroblast contraction of floating collagen gel: interaction between epigallocatechin-3-O-gallate and platelet derived growth factor.
The effects of (-)-epigallocatechin gallate (EGCG) on the contraction of floating collagen gel by fibroblasts were investigated. EGCG inhibited collagen gel contraction dose-dependently. On the basis of the fact that platelet-derived growth factor (PDGF) is one of the serum components with stimulatory activity in collagen gel contraction, we examined the possibility that interaction between EGCG and PDGF may be involved in this inhibition mechanism. We confirmed this by recombinant PDGF-BB in the present system and we found that EGCG inhibited PDGF-stimulated collagen gel contraction. The results of affinity chromatography indicated that PDGF was bound by EGCG immobilized on agarose gel as detected by enzyme-linked immunoassay and Western blotting. These findings suggest that binding of EGCG to PDGF is at least partly involved in the mechanism of inhibition of collagen gel contraction by EGCG.


Estrogen genotoxicity causes preferential development of Fuchs endothelial corneal dystrophy in females.
Fuchs endothelial corneal dystrophy (FECD) is a genetically complex, age-related, female-predominant disorder characterized by loss of post-mitotic corneal endothelial cells (CEnCs). Ultraviolet-A (UVA) light has been shown to recapitulate the morphological and molecular changes seen in FECD to a greater extent in females than males, by triggering CYP1B1 upregulation in females. Herein, we investigated the mechanism of greater CEnC susceptibility to UVA in females by studying estrogen metabolism in response to UVA in the cornea. Loss of NAD(P)H quinone oxidoreductase 1 (NQO1) resulted in increased production of estrogen metabolites and mitochondrial-DNA adducts, with a higher CEnC loss in Nqo1-/- female compared to wild-type male and female mice. The CYP1B1 inhibitors, trans-2,3',4,5'-tetramethoxystilbene (TMS) and berberine, rescued CEnC loss. Injection of wild-type male mice with estrogen (E2; 17β-estradiol) increased CEnC loss, followed by increased production of estrogen metabolites and mitochondrial DNA (mtDNA) damage, not seen in E2-treated Cyp1b1-/-male mice. This study demonstrates that the endo-degenerative phenotype is driven by estrogen metabolite-dependent CEnC loss that is exacerbated in the absence of NQO1; thus, explaining the mechanism accounting for the higher incidence of FECD in females. The mitigation of estrogen-adduct production by CYP1B1 inhibitors could serve as a novel therapeutic strategy for FECD.


Dietary cocoa ameliorates non-alcoholic fatty liver disease and increases markers of antioxidant response and mitochondrial biogenesis in high fat-fed mice.
Cocoa powder, derived Theobroma cacao, is a popular food ingredient that is commonly consumed in chocolate. Epidemiological and human intervention studies have reported that chocolate consumption is associated with reduced risk of cardiometabolic diseases. Laboratory studies have reported the dietary supplementation with cocoa or cocoa polyphenols can improve obesity and obesity-related comorbidities in preclinical models. Non-alcoholic fatty liver disease (NAFLD), one such comorbidity, is a risk factor for cirrhosis and hepatocellular carcinoma. Limited studies have examined the effect of cocoa/chocolate on NAFLD and underlying hepatoprotective mechanisms. Here, we examined the hepatoprotective effects of dietary supplementation with 80 mg/g cocoa powder for 10 wks in high fat (HF)-fed obese male C57BL/6J mice. We found that cocoa-supplemented mice had lower rate of body weight gain (22%), hepatic triacylglycerols (28%), lipid peroxides (57%), and mitochondrial DNA damage (75%) than HF-fed controls. These changes were associated with higher hepatic superoxide dismutase and glutathione peroxidase enzyme activity and increased expression of markers of hepatic mitochondrial biogenesis. We also found that the hepatic protein expression of sirtuin 3 (SIRT3), and mRNA expression of peroxisome proliferator activated receptor g coactivator (PGC) 1a, nuclear respiratory factor 1, and forkhead box O3 were higher in cocoa-treated mice compared to HF-fed controls. These factors play a role in coordinating mitochondrial biogenesis and expression of mitochondrial antioxidant response factors. Our results indicate that cocoa supplementation can mitigate the severity of NAFLD in obese mice and that these effects are related to SIRT3/PGC1a-mediated increases in antioxidant response and mitochondrial biogenesis.


Impaired GABAergic inhibition in the visual cortex of brain-derived neurotrophic factor heterozygous knockout mice.
Brain derived neurotrophic factor (BDNF) promotes the formation, maturation and stabilization of inhibitory synapses in the central nervous system. In addition, BDNF has been suggested to regulate the critical period for ocular dominance plasticity in the visual system. Here we further evaluated the role of BDNF in the visual cortex by studying the GABAergic synaptic transmission under conditions of chronically reduced levels of BDNF. Whole-cell patch-clamp recordings were performed from pyramidal neurons located in layers II/III of visual cortical slices in heterozygous BDNF knockout mice (BDNF (+/-)) and their wild-type littermates at the age of 21-25 days. The BDNF (+/-) mice showed a decreased frequency and amplitude of miniature inhibitory postsynaptic currents (mIPSCs) as well as a reduced amplitude and prolonged decay time constant of evoked IPSCs. Further analyses indicated an impaired presynaptic GABAergic function in BDNF (+/-) mice, as shown by the decreased release probability, steady-state release and synchronous release of GABA. However, the number of functional release sites remained unchanged. In line with these observations, an impaired glutamate-driven GABA release was observed in BDNF (+/-) mice. Furthermore, the overall balance in the strength of cortical excitation to inhibition shifted towards a decreased inhibition. Finally, the reversal potential for chloride-mediated evoked IPSCs was not affected. These findings suggested that chronically reduced levels of BDNF strongly impair the GABAergic inhibitory function in visual cortex by altering postsynaptic properties and by reducing presynaptic GABA release as well as the overall strength of inhibition onto pyramidal neurons within the cortical network. These impairments of inhibitory function are compatible with a rather immature status of the GABAergic system in BDNF (+/-) mice, which supports the hypothesis that the level of expression for BDNF critically affects maturation and function of the GABAergic inhibition.


Acetate metabolism in a pta mutant of Escherichia coli W3110: importance of maintaining acetyl coenzyme A flux for growth and survival.
In order to study the physiological role of acetate metabolism in Escherichia coli, the growth characteristics of an E. coli W3100 pta mutant defective in phosphotransacetylase, the first enzyme of the acetate pathway, were investigated. The pta mutant grown on glucose minimal medium excreted unusual by-products such as pyruvate, D-lactate, and L-glutamate instead of acetate. In an analysis of the sequential consumption of amino acids by the pta mutant growing in tryptone broth (TB), a brief lag between the consumption of amino acids normally consumed was observed, but no such lag occurred for the wild-type strain. The pta mutant was found to grow slowly on glucose, TB, or pyruvate, but it grew normally on glycerol or succinate. The defective growth and starvation survival of the pta mutant were restored by the introduction of poly-beta-hydroxybutyrate (PHB) synthesis genes (phbCAB) from Alcaligenes eutrophus, indicating that the growth defect of the pta mutant was due to a perturbation of acetyl coenzyme A (CoA) flux. By the stoichiometric analysis of the metabolic fluxes of the central metabolism, it was found that the amount of pyruvate generated from glucose transport by the phosphoenolpyruvate-dependent phosphotransferase system (PTS) exceeded the required amount of precursor metabolites downstream of pyruvate for biomass synthesis. These results suggest that E. coli excretes acetate due to the pyruvate flux from PTS and that any method which alleviates the oversupply of acetyl CoA would restore normal growth to the pta mutant.


Enhanced plasmonic photovoltaic using embedded novel gear-shaped nanoparticles.
In this paper, novel gear-shaped nanoparticles are introduced for the first time to enhance the photovoltaic (PV) efficiency. This has been achieved via increasing the overall power absorption by the PV semiconductor material in both visible and near-infrared ranges. The modes of the new gear-shaped nanoparticles are investigated. A parametric study has been performed which demonstrates how the design parameters of the proposed nanoparticles can be engineered for best overall power absorption within a Si surrounding medium. A figure of merit (FoM) is defined that takes into account all objectives. An optimization technique is applied to obtain the optimum set of the gear's dimensions, penetration depth, and periodicity for the maximum possible FoM. The optimum gear-shaped nanoparticles design offers 48% enhancement in the FoM if compared with a bare Si block with no nanoparticles and 7% enhancement over the conventional disk-shaped nanoparticles. The enhancement gained by the embedded gear-shaped nanoparticles on the J-V characteristics of the PV is also studied, and the effects of changing the dimensions and the position of nanoparticles on the J-V characteristics enhancement are investigated.


Discovery of sinomenine/8-Bis(benzylthio)octanoic acid hybrids as potential anti-leukemia drug candidate via mitochondrial pathway.
Traditional Chinese medicine Qingfengteng primarily acquired from the dried canes of Sinomenium acutum (Thunb.) Rehd. et Wils. var. cinereum Rehd. et Wils. and S. acutum (Thunb.) Rehd. et Wils. For the therapeutic treatment of rheumatism, acute arthritis, and rheumatoid arthritis based on Qingfengteng, sinomenine hydrochloride was recently made the principal active ingredient in various dosage forms. 8-Bis(benzylthio)octanoic acid (CPI-613) was an orphan medicine that the FDA and EMA approved orphan for the treatment of certain resistant malignancies. Its unique mode of action and minimal toxicity toward normal tissues made for an apt pharmacophore. In order to expand the field of sinomenine anticancer structures, sinomenine/8-Bis(benzylthio)octanoic acid derivatives were designed and synthesized. Among them, target hybrids e4 stood out for having notable cytotoxic effects against cancer cell lines, especially for K562 cells, with IC50 values of 2.45 μM and high safety. In-depth investigations demonstrated that e4 caused apoptosis by stopping the cell cycle at G1 phase, and doing so by altering the morphology of the nucleus and causing membrane potential of the in mitochondria to collapse. These results indicated e4 exerted an antiproliferative effect through apoptosis induction via mitochondrial pathway.


X-ray magnetic resonance fusion to internal markers and utility in congenital heart disease catheterization.
X-ray magnetic resonance fusion (XMRF) allows for use of 3D data during cardiac catheterization. However, to date, technical requirements have limited the use of this modality in clinical practice. We report on a new internal-marker XMRF method that we have developed and describe how we used XMRF during cardiac catheterization in congenital heart disease. XMRF was performed in a phantom and in 23 patients presenting for cardiac catheterization who also needed cardiac MRI for clinical reasons. The registration process was performed in < 5 minutes per patient, with minimal radiation (0.004 to 0.024 mSv) and without contrast. Registration error was calculated in a phantom and in 8 patients using the maximum distance between angiographic and 3D model boundaries. In the phantom, the measured error in the anteroposterior projection had a mean of 1.15 mm (standard deviation, 0.73). The measured error in patients had a median of 2.15 mm (interquartile range, 1.65 to 2.56 mm). Internal markers included bones, airway, image artifact, calcifications, and the heart and vessel borders. The MRI data were used for road mapping in 17 of 23 (74%) cases and camera angle selection in 11 of 23 (48%) cases. Internal marker-based registration can be performed quickly, with minimal radiation, without the need for contrast, and with clinically acceptable accuracy using commercially available software. We have also demonstrated several potential uses for XMRF in routine clinical practice. This modality has the potential to reduce radiation exposure and improve catheterization outcomes.


Telomere maintenance in tumour cells.
Telomere attrition may regulate the proliferative life span of somatic cells and contribute to the genetic instability of tumour cells, and telomere maintenance is required for cell survival. The ample, but mainly correlative, evidence in support of these hypotheses is now beginning to be complemented by experimental results. There have also been unexpected findings that attest to the complexity of the biological processes and systems under study. Mammalian telomere biology has definitely entered into an exciting phase. Given the increasing pace of research on this topic, it seems most likely that a not too distant future will provide us with answers to many of the unresolved issues mentioned in this article.


Complementation of the movement-deficient mutations in potato virus X: potyvirus coat protein mediates cell-to-cell trafficking of C-terminal truncation but not deletion mutant of potexvirus coat protein.
The cell-to-cell movement of the GUS-tagged potato virus X (PVX) coat protein (CP) movement-deficient mutant was restored by potyviral CPs of potato virus A (PVA) and potato virus Y (PVY) in Nicotiana benthamiana leaves in transient cobombardment experiments. Viral cell-to-cell movement of PVX CP mutant was complemented in Nicotiana tabacum cv. SR1 transgenic plants expressing PVY CP: PVX RNA and polymerase were detected in the PVX CP mutant-inoculated leaves of transgenic plants. These findings demonstrated the ability of the PVX CP-deficient mutant to move from cell to cell but not long distances in the transgenic plants and suggest that CPs of potex- and potyviruses display complementary activities in the movement process. Potyviral CP alone is not able to carry out these activities, since the mutated PVX CP is indispensable for restored movement. No trans-encapsidation between potyviral CP and PVX RNA was observed. Therefore, potyviral CP facilitates the PVX CP mutant movement by the mechanism that cannot be explained by coat protein substitution. Our data also suggest that CP functioning in cell-to-cell movement is not restricted to a simple passive role in forming virions.


Effects of interferon-gamma on neuronal infections.
IFN-gamma is released in the brain, not only during acute infectious diseases or immunological reactions, but also for extended periods of time after clearance of infectious virus and during viral latency. In this review, we focus on the role of IFN-gamma in the interaction between a neuron and a pathogen, that is, the role in implementation of microbial destruction, stasis or persistence, its potential neuroprotective or toxic effects, and how this cytokine may affect neuronal function and the behavior of an individual.


Transdermal nicotine use in postmenopausal women: does the treatment efficacy differ in women using and not using hormone replacement therapy?
This study of postmenopausal female smokers (N = 94) asked: During short-term smoking abstinence, do the beneficial effects of transdermal nicotine replacement therapy (NRT) on acute symptomatology (i.e., withdrawal, cigarette craving, smoking urges, mood, depressive symptoms, motor speed, and reaction time) differ in women who use and do not use hormone replacement therapy (HRT)? Participants were recruited according to HRT and non-HRT use (self-selecting), then randomized within strata to active nicotine or placebo nicotine patch. After 1 baseline week of smoking, participants quit smoking for 2 weeks. Women received cessation counseling and were monitored for abstinence. Dependent measures were collected during five clinic visits. Two-way analysis of covariance (ANCOVA) were run on change scores for dependent variables, with nicotine patch group (active/placebo) and HRT group (HRT/non-HRT) as independent variables and age as a covariate. No interactions were found between HRT and patch condition, but both showed specific effects. During the first abstinent week, women on active nicotine patch (compared with placebo) experienced less severe withdrawal, greater reductions in cigarette cravings, and lower (more favorable) Factor 1 scores on the Questionnaire of Smoking Urges. During the second abstinent week, women using HRT (compared with the non-HRT group) exhibited better mood (Profile of Mood States scores) and less depression (Beck Depression Inventory scores). These results suggest the following: First, the efficacy of transdermal nicotine replacement is not adversely modified by women's HRT use; second, ovarian hormones might influence women's responses to smoking cessation, and thus should be considered in developing effective strategies for women to quit smoking.


The Role of p53 in HIV Infection.
This review aims to elucidate the multifaceted role of the tumor suppressor protein p53 in the context of HIV infection. We explore how p53, a pivotal regulator of cellular processes, interacts with various facets of the HIV life cycle. Understanding these interactions could provide valuable insights into potential therapeutic interventions and the broader implications of p53 in viral infections. Recent research has unveiled a complex interplay between p53 and HIV. Several reports have highlighted the involvement of p53 in restricting the replication of HIV within both immune and nonimmune cells. Various mechanisms have been suggested to unveil how p53 enforces this restriction on HIV replication. However, HIV has developed strategies to manipulate p53, benefiting its replication and evading host defenses. In summary, p53 plays a multifaceted role in HIV infection, impacting viral replication and disease progression. Recent findings underscore the importance of understanding the intricate interactions between p53 and HIV for the development of innovative therapeutic approaches. Manipulating p53 pathways may offer potential avenues to suppress viral replication and ameliorate immune dysfunction, ultimately contributing to the management of HIV/AIDS. Further research is warranted to fully exploit the therapeutic potential of p53 in the context of HIV infection.


Assessment of dental caries prevention program applied to a cohort of elementary school children of Kebemer, a city in Senegal.
Dental caries is frequently observed in children, particularly among those residing in developing countries. The most adapted strategies against this pathology remains prevention based on information, education, and communication (IEC), as well as on early diagnosis and treatment. We carried out a study that aimed to analyze the development of dental caries in a cohort of school children followed during their primary education. The objective was to assess the evolution of the dental status of a cohort of students during their elementary curriculum. A cohort of school children was followed during 6 years from the first grade to the sixth grade. Monitoring of these school children focused every year on IEC based on learning methods of brushing messages, dietary advice, systematic visits, fluoride use, and primary dental care. During the school year, the students were periodically subjected to education and communication briefings (IEC). Primary care consisted of extracting and descaling rhizalyzed teeth in the same period. The data from this review were collected using the World Health Organization questionnaire, and statistical analysis was performed with the software Epi-info version 6.04 d. The mean age of the 171 school children was 6 years in the first grade and 11 years in the sixth grade. In the first grade, the decayed permanent teeth prevalence was 31.6% and the In permanent teeth: Decayed, missing or filled teeth (DMF/T) was 0.47. The decayed primary teeth prevalence was 75% and the in primary teeth: decayed or filled teeth (df/t) 2.23. In the sixth year, the prevalence of decayed permanent teeth was 51% and DMF/T 0.36 whereas the decayed primary teeth prevalence was 12% and the df/t was 0.19. The prevalence of decayed permanent teeth increased from 31.6 to 51% whereas the mean DMF/T was not statistically different between school children of the first and sixth grade class. The promotion of oral health by IEC messages and monitoring of children constitute an effective preventive method for children's oral care.


Association of Cigarette Smoking Patterns Over 30 Years With Audiometric Hearing Impairment and Speech-in-Noise Perception: The Atherosclerosis Risk in Communities Study.
The implications of cigarette smoking and smoking cessation for hearing impairment remain unknown. Many studies on this topic have failed to account for attrition among smokers in their findings. To assess the association of cigarette smoking patterns with audiometric and speech-in-noise hearing measures among participants of the Atherosclerosis Risk in Communities Study. This cross-sectional study included participants of the Atherosclerosis Risk in Communities Study from 4 US communities. The analysis includes data from visit 1 (1987-1989) through visit 6 (2016-2017); data were analyzed from March 16 through June 25, 2021. Audiometric hearing and speech-in-noise testing was offered to all participants at visit 6. Participants with incomplete audiometric data or missing data for educational level, body mass index, drinking status, a diabetes or hypertension diagnosis, or occupational noise were excluded. In addition, individuals were excluded if they self-reported as having other than Black or White race and ethnicity, or if they self-reported as having Black race or ethnicity and lived in 2 predominantly White communities. Smoking behavior was classified from visit 1 (1987-1989) to visit 6 (2016-2017) using group-based trajectory modeling based on self-reported smoking status at each clinic visit. Hearing was assessed at visit 6. An audiometric 4-frequency (0.5, 1, 2, 4 kHz) pure-tone average (PTA) was calculated for the better-hearing ear and modeled as a continuous variable. Speech-in-noise perception was assessed via the Quick Speech-in-Noise Test (QuickSIN) and modeled continuously. Attrition during the 30 years of follow-up was addressed by inverse probability of attrition weighting. A total of 3414 participants aged 72 to 94 years (median [IQR] age, 78.8 [76.0-82.9] years; 2032 [59.5%] women) when hearing was measured at visit 6 (2016-2017) were included in the cohort; 766 (22.4%) self-identified as Black and 2648 (77.6%) as White individuals. Study participants were classified into 3 smoking groups based on smoking behavior: never or former smoking at baseline (n = 2911 [85.3%]), quit smoking during the study period (n = 368 [10.8%]), and persistent smoking (n = 135 [4.0%]). In fully adjusted models, persistent smoking vs never or former smoking was associated with an average 2.69 (95% CI, 0.56-4.81) dB higher PTA (worse hearing) and 1.42 (95% CI, -2.29 to -0.56) lower QuickSIN score (worse performance). Associations were stronger when accounting for informative attrition during the study period (3.53 [95% CI, 1.14-5.93] dB higher PTA; 1.46 [95% CI, -2.52 to -0.41] lower QuickSIN scores). Smoking cessation during the study (vs never or former smoking) was not associated with changes in hearing. In this cross-sectional study, persistent smoking was associated with worse audiometric hearing and speech-in-noise perception. Hearing measures among participants who quit smoking during the study period did not differ from those for never or former smokers, indicating that smoking cessation (as opposed to persistent smoking) may have benefits for hearing health.


Chemical composition and antibacterial activity of essential oil of Heracleum rechingeri Manden from Iran.
The chemical composition of the essential oil from Heracleum rechingeri was analysed using GC-MS. Twenty-seven compounds, accounting for 94.62% of the extracted essential oil, were identified. The main oil compounds were octyl acetate (29.49%), elemicine (23.06%), (E)-caryophyllene (9.26%), caryophyllene oxide (6.42%), terpinolene (6.12%) and (Z)-3-octenyl acetate (4.72%). The antimicrobial activity of the essential oil was tested against three different bacteria by disc diffusion method and showed maximum inhibitory activity against Gram-positive bacteria, especially Bacillus subtilis.


Virtual and dynamic hierarchical architecture: an overlay network topology for discovering grid services with high performance.
This paper presents an overlay network topology called Virtual and Dynamic Hierarchical Architecture (VDHA) for discovering Grid services with high performance. Service discovery based on VDHA have scalable, autonomous, efficient, reliable and quick responsive. We propose two service discovery algorithms. Full Search Query and Discovery Protocol (FSQDP) discovers the nodes that match the request message from all N nodes, which has time complexity O(logN), space complexity O(nvg) (nvg being node numbers of each virtual group), and message-cost O(N), and Domain-Specific Query and Discovery Protocol (DSQDP) searches nodes in only specific domains with time complexity O(nvg), space complexity O(nvg), and message-cost O(nvg). In this paper, we also describe VDHA, its formal definition, and Grid Group Management Protocol.


Primary malignant melanoma of the esophagus.
Primary malignant melanoma of the esophagus (PMME) comprises only 0.1-0.2% of all malignant esophageal tumors. PMME tumors are highly aggressive and metastasize early via hematogenic and lymphatic pathways. Treatment outcome is poor because the cancer has often advanced at the time of diagnosis. Inoperability, unsuccessful treatment with radiotherapy and chemotherapy in advanced tumors and metastases have contributed to its poor prognosis. Here, we present the endoscopic features, endoscopic ultrasonography findings and management of a PMME case.


Clinical and radiographic success of (partial) pulpotomy and pulpectomy in primary teeth: A systematic review.
The aim of this systematic review was to evaluate the clinical outcome of partial pulpotomy, pulpotomy and pulpectomy for treating primary teeth with normal or infected pulp or with irreversible pulpitis. Two reviewers on Pubmed and ISI Web of Science performed a comprehensive literature review of publications from 1966 until July 2019. Pico outline was used to facilitate literature research. Among abstracts, publications were selected according to the following criteria: prospective clinical study, correct indication for the performed treatment, clear definition of clinical and/or radiographic success criteria and at least 6-month follow-up period. The strict selection criteria under the keywords "pulpotomy", "partial pulpotomy" and "pulpectomy" resulted in a limited amount of randomised controlled trials (RCT) or controlled clinical trials (CT). Qualitative assessment of the selected clinical studies and level of evidence was included according to the criteria described by the Oxford Centre for Evidence-Based Medicine (CEBM). Prerequisites for a successful pulpotomy are symptom-free teeth, sterile removal of coronal pulp and haemostasis. Both MTA and formocresol perform well for partial pulpotomies after caries exposure. Formocresol had been the most popular amputation material for pulpotomies. Due to the potential side effects, other medicaments, such as ferric sulfate, mineral trioxide aggregate (MTA) or NaOCl are suggested. Grey and white MTA yeld the same results. Lasers are not recommended due to their large diversity. Regarding pulpectomy, the conditions, procedures, and evaluation for the treatment were not well defined in the studies. Nevertheless, there is evidence to use calcium hydroxide, zinc oxide eugenol paste or iodoform based pastes as root filling materials for non-vital molars. Pulpectomies showed better success rates than pulpotomies. Stainless steel crowns are recommended as definite restorations after both endodontic treatments. Longer follow-up periods, further clinical studies with comparable conditions and clear definition of evaluation criteria are needed to further confirm the results of endodontic treatment in primary teeth.


Bacterial community dynamics in high and low bioavailability soils following laboratory exposure to a range of hexahydro-1,3,5-trinitro-1,3,5-triazine concentrations.
Few studies have examined the potential long-term effects of high concentrations of hexahydro-1,3,5-trinitro-1,3,5-triaxine (RDX) on bacterial communities in soil. In the present study, a sandy loam soil and a silt loam soil (high and low bioavailability, respectively) were artificially contaminated with RDX (0, 50, 500, 1,500, 5,000, 10,000, and 15,000 mg/kg soil). Bacterial communities from each treatment were monitored over 63 d to characterize the effects of RDX exposure on bacterial activity, biomass, functional diversity (Biolog microtiter plates), and structural diversity (denaturing gradient gel electrophoresis of 16S rDNA). Bacterial communities native to the high bioavailability soil were inherently different than bacterial communities native to the silt loam soil, not only in terms of bacterial activity and biomass, but also in terms of bacterial community functional and structural diversity. Soil RDX contamination was correlated with decreased bacterial biomass in the silt loam soil treatments and with decreased bacterial activity in the sandy loam soil treatments on day 7. Soil RDX contamination did not cause a significant shift in the functional diversity of the bacterial communities native to the silt loam soil, but was correlated with a shift in identities of substrates used by bacterial communities native to the sandy loam soil on day 7. Bacterial community structure was insensitive to the gradient of RDX concentrations at the beginning of the incubation. However, the identities of carbon substrates used by bacterial communities in both soil types were affected by long-term incubation with RDX.


Thymoquinone-loaded self-nano-emulsifying drug delivery system against ischemia/reperfusion injury.
In the present study, a self-nano-emulsifying drug delivery system (SNEDDS) was developed to evaluate the efficiency of thymoquinone (TQ) in hepatic ischemia/reperfusion. SNEDDS was pharmaceutically characterized to evaluate droplet size, morphology, zeta potential, thermodynamic stability, and dissolution/diffusion capacity. Animals were orally pre-treated during 10 days with TQ-loaded SNEDDS. Biochemical analyses, hematoxylin-eosin staining, indirect immunofluorescence, and reverse transcription polymerase chain reaction (RT-PCR) were carried out to assess cell injury, oxidative stress, inflammation, and apoptosis. The TQ formulation showed good in vitro characteristics, including stable nanoparticle structure and size with high drug release rate. In vivo determinations revealed that TQ-loaded SNEDDS pre-treatment of rats maintained cellular integrity by decreasing transaminase (ALT and AST) release and preserving the histological characteristics of their liver. The antioxidant ability of the formulation was proven by increased SOD activity, reduced MDA concentration, and iNOS protein expression. In addition, this formulation exerted an anti-inflammatory effect evidenced by reduced plasma CRP concentration, MPO activity, and gene expressions of TLR-4, TNF-α, NF-κB, and IL-6. Finally, the TQ-loaded SNEDDS formulation promoted cell survival by enhancing the Bcl-2/Bax ratio. In conclusion, our results indicate that TQ encapsulated in SNEDDS significantly protects rat liver from I/R injury.


A small nuclear acidic protein (MTI-II, Zn2+-binding protein, parathymosin) attenuates TNF-α inhibition of BMP-induced osteogenesis by enhancing accessibility of the Smad4-NF-κB p65 complex to Smad binding element.
Pro-inflammatory cytokines prevent bone regeneration in vivo and activation of nuclear factor-κB (NF-κB) signaling has been proposed to lead to suppression of bone morphogenetic protein (BMP)-induced osteogenesis via direct binding of p65 to Smad4 in vitro. Application of a small nuclear acidic protein (MTI-II) and its delivered peptide, MPAID (MTI-II peptide anti-inflammatory drug) has been described to elicit therapeutic potential via strong anti-inflammatory action following the physical association of MTI-II and MPAID with p65. However, it is unclear whether MTI-II attenuates tumor necrosis factor (TNF)-α inhibition of BMP-induced osteogenesis. Herein, we found that TNF-α-mediated suppression of responses associated with BMP4-induced osteogenesis, including expression of the osteocalcin encoding gene Ocn, Smad binding element (SBE)-dependent luciferase activity, alkaline phosphatase activity, and alizarin red S staining were largely restored by MTI-II and MPAID in MC3T3-E1 cells. Mechanistically, MTI-II and MPAID did not inhibit nuclear translocation of p65 or disassociate Smad4 from p65. Further, results from chromatin immunoprecipitation (ChIP) analyses revealed that Smad4 enrichment in cells over-expressing MTI-II and treated with TNF-α was equivalent to that in cells without TNF-α treatment. Alternatively, Smad4 enrichment was considerably decreased following TNF-α treatment in control cells. Moreover, p65 enrichment in the Id-1 promoter SBE was detected only when cells over-expressing MTI-II were stimulated with TNF-α. Overall, our study concludes that MTI-II restored TNF-α-inhibited suppression of BMP-Smad-induced osteogenic differentiation by enhancing accessibility of the Smad4-p65 complex to the SBE rather than by liberating Smad4 from p65.


A photodynamic antibacterial spray-coating based on the host-guest immobilization of the photosensitizer methylene blue.
In the "post-antibiotic era", healthcare-associated infection has become a global problem that threatens public health and causes huge economic losses. The development of antibacterial coatings based on non-antibiotic strategies is particularly important as drug-resistant bacteria continue to evolve. Photodynamic coatings are a high potential method to treat bacteria, however, the aggregation of photosensitizers on the coating affects the photodynamic capacity seriously. Herein, a photodynamic coating is developed based on the host-guest interaction between β-cyclodextrin and the photosensitizer methylene blue (MB). The host-guest interaction avoids aggregation of MB and results in a high singlet oxygen quantum yield. Consequently, efficient photoantibacterial activity towards methicillin-resistant Staphylococcus aureus is achieved by the photodynamic coating with very low MB density (0.53 ± 0.06 μg cm-2).


Invited article: comparative effectiveness research, evidence-based medicine, and the AAN.
Comparative effectiveness research (CER) is the study of the relative effects of treatments to determine which will be most likely to improve overall health for a specific condition. This area has received a great deal of political focus, and substantial funding for CER is included in the American Reinvestment and Recovery Act of 2009. The results of CER are intended to inform evidence-based guidelines and to improve the quality and effectiveness of medical care. In the absence of such research, guidelines often depend on consensus to rank available therapies. We believe that an increase in CER would clearly enhance evidence-based guidelines. However, the research must be performed and analyzed with great care to avoid reaching unhelpful, or even harmful, conclusions. Specifically, individual patient characteristics must be taken into account, study endpoints must approximate the most important patient outcomes, therapies must be used optimally within the studies, and the most relevant therapies for a given indication must be included for comparison. CER that is not performed or interpreted correctly could have the potential to affect negatively our choices of therapies. The neurology community must help inform the process of CER to ensure the highest-quality research, which in turn will result in the most valid outcomes.


Circulating interleukin-1 receptor antagonist (IL-1RA) serum levels in patients undergoing orthotopic heart transplantation.
In a pilot study we determined the serum levels of circulating interleukin-1 receptor antagonist (IL-1ra) in patients undergoing orthotopic heart transplantation and in control patients scheduled for open heart surgery without allograft transplantation. Blood samples were obtained from 12 transplant recipients and 7 controls prior to the operative procedures to determine baseline values. Serum levels of IL-1ra were measured within 12 h of decrossclamping of the aorta and every 24 h for the following 14 days. Endomyocardial biopsies were obtained weekly for the 1st month after transplantation. Compared to baseline values, IL-1ra serum levels 12 h after decrossclamping of the aorta were significantly higher both in the control group (507 +/- 165 vs 3980 +/- 452 pg/ml, P < 0.01) and among the transplant recipients (413 +/- 180 vs 4117 +/- 459 pg/ml, P < 0.01) IL-1ra levels remained significantly elevated for 2 and 5 days, respectively. There were no significant differences in the IL-1ra serum levels between the two groups throughout the observation period. Endomyocardial biopsies of two patients showed acute allograft rejection, Billingham grade IIIa and IIIb, respectively. In both cases, the rejection episodes were accompanied by a renewed and more pronounced elevation in the IL-1ra serum levels beyond 4000 pg/ml for at least 2 days. These preliminary results indicate that IL-1ra may be a nonspecific immune marker during the first few days after orthotopic heart transplantation and cardiopulmonary bypass. Moreover, renewed, prolonged increases in IL-1ra appear to be associated with rejection. Further studies are needed to confirm the predictive value of IL-1ra in the detection of acute allograft rejection.


Mucosal immunotherapy with CpG oligodeoxynucleotides reverses a murine model of chronic asthma induced by repeated antigen exposure.
Murine models of acute atopic asthma may be inadequate to study the effects of recurrent exposure to inhaled allergens, such as the epithelial changes seen in asthmatic patients. We developed a murine model in which chronic airway inflammation is maintained by repeated allergen [ovalbumin (OVA)] inhalation; using this model, we examined the response to mucosal administration of CpG DNA (oligonucleotides) and specific antigen immunotherapy. Mice repeatedly exposed to OVA developed significantly greater airway hyperresponsiveness and goblet cell hyperplasia, but not airway eosinophilia, compared with those exposed only twice. CpG-based immunotherapy significantly reversed both acute and chronic markers of inflammation as well as airway hyperresponsiveness. We further examined the effect of mucosal immunotherapy on the response to a second, unrelated antigen. Mice sensitized to both OVA and schistosome eggs, challenged with inhaled OVA, and then treated with OVA-directed immunotherapy demonstrated significant reduction of airway hyperresponsiveness and a moderate reduction in eosinophilia, after inhalation challenge with schistosome egg antigens. In this model, immunotherapy treatment reduced bronchoalveolar lavage (BAL) levels of Th2 cytokines (IL-4, IL-5, IL-13, and IL-10) without changing BAL IFN-gamma. Antigen recall responses of splenocytes from these mice demonstrated an antigen-specific (OVA) enhanced release of IL-10 from splenocytes of treated mice. These results suggest that CpG DNA may provide the basis for a novel form of immunotherapy of allergic asthma. Both antigen-specific and, to a lesser extent, antigen-nonspecific responses to mucosal administration of CpG DNA are seen.


Loving yourself abundantly: relationship of the narcissistic personality to self- and other perceptions of workplace deviance, leadership, and task and contextual performance.
The authors report results from 2 studies assessing the extent to which narcissism is related to self- and other ratings of leadership, workplace deviance, and task and contextual performance. Study 1 results revealed that narcissism was related to enhanced self-ratings of leadership, even when controlling for the Big Five traits. Study 2 results also revealed that narcissism was related to enhanced leadership self-perceptions; indeed, whereas narcissism was significantly positively correlated with self-ratings of leadership, it was significantly negatively related to other ratings of leadership. Study 2 also revealed that narcissism was related to more favorable self-ratings of workplace deviance and contextual performance compared to other (supervisor) ratings. Finally, as hypothesized, narcissism was more strongly negatively related to contextual performance than to task performance.


The effects of the statins lovastatin and cerivastatin on signalling by the prostanoid IP-receptor.
The prostanoid-IP receptor may be unique among G protein coupled receptors in that it is isoprenylated. In this study, we investigated the effects of the statins lovastatin and cerivastatin on signalling by the mouse (m) IP and the human (h) IP receptors, over-expressed in human embryonic kidney (HEK) 293 cells and by the hIP receptor, endogenously expressed in human erythroleukaemia cells. Both statins significantly reduced IP receptor-mediated cyclic AMP generation and intracellular calcium ([Ca(2+)](i)) mobilization in a time and concentration dependent manner but had no effect on signalling by the non-isoprenylated beta(2) adrenergic receptor or by the human prostanoid-TP receptor isoforms. Cerivastatin (IC(50), 50 - 90 nM) was significantly more potent than lovastatin (IC(50), 0.80 - 4.2 microM) in inhibiting IP receptor signalling. Whereas IC(50) values indicated that the hIP receptor was significantly more sensitive than the mIP receptor to the statins, the extent of inhibition of cyclic AMP generation by the mIP receptor was significantly greater than that of the hIP receptor to either statin, even at the highest concentrations used. Pretreatment with either statin significantly reduced IP receptor mediated desensitization of signalling by the h.TPalpha, but not by the h.TPbeta, receptor isoform. These data generated in whole cells point to the possibility that statin therapy may interfere with IP receptor signalling in vivo; such interference may be extenuated under conditions where circulating statin levels are elevated and may account, in part, for some of the pleiotropic affects of the statins not attributed solely to their lipid lowering properties.


Bone preservation with a conservative metaphyseal loading implant.
Proximal load transfer and the absence of distal fixation of the stem are crucial to obtain the best behaviour of the femoral bone after total hip replacement. In this study, dual energy X-ray absorptiometry (DEXA) was employed to understand and compare the bone density changes and thus the re-distribution of mechanical forces in two different extra short stems. Two cohorts of ten patients were included in this retrospective study. Both implants are custom-made and present a well defined lateral flare. The first model is fully coated and presents a short stem (Group A), the second is an unstemmed metaphyseal implant with a polished tip (Group B). DEXA scans were obtained in all patients at the two-year follow-up. A higher BMD was detected in ROIs 1, 2, 4, 5 in Group B confirming a preservation of the proximal bone mass and thus indirectly a more proximal load transfer. The results obtained confirm the importance of the geometry of the implant on proximal bone density. Loading both medial and lateral proximal femoral flares and the complete absence of the diaphyseal portion of the stem provide the optimal bone remodelling of the femur after total hip replacement.


Trend of burnout among Swiss doctors.
Over the last decade the Swiss health care system has undergone several changes, resulting in stronger economic constraints, a heavier administrative workload and limited work autonomy for doctors. In this context we examined the change in burnout prevalence over time among Swiss doctors surveyed during this period. Cross-sectional survey data collected by mail in 2002, 2004 and 2007 throughout the country were used. Measures included the Maslach Burnout Inventory (MBI), several socio-demographics (gender, living alone, having children), and work-related characteristics (number of years in current workplace, hours worked). Answers to the MBI were used to classify respondents into moderate (high score on either the emotional exhaustion or the depersonalisation/cynicism subscale) and high degree of burnout (scores in the range of burnout in all three scales). Rates of moderate-degree burnout increased from 33% to 42% among general practitioners (p = 0.002) and from 19% to 34% among paediatricians (p = 0.001) (high degree of burnout: 4% to 6% [p = 0.17] and 2% to 4% [p = 0.42] respectively). After adjustment for significant socio-demographic and work-related characteristics, an increased risk of moderate burnout was found for doctors surveyed in 2004 and 2007 (OR 1.6, 95%CI 1.3 to 2.0), general practitioners (OR 1.6, 95%CI 1.3 to 2.0) and French-speaking doctors (OR 1.5, 95%CI 1.3 to 1.9). An increased risk of high-degree burnout was found only for general practitioners (OR 1.8, 95%CI 1.05 to 3.0). Burnout levels among Swiss doctors have increased over the last decade, in particular among French-speaking doctors.


ABCB6 mRNA and DNA methylation levels serve as useful biomarkers for prediction of early intrahepatic recurrence of hepatitis C virus-related hepatocellular carcinoma.
The poor prognosis of hepatocellular carcinoma (HCC) can be explained largely by the high rate of intrahepatic recurrence (IHR). Identification of genes related to IHR is needed to improve the poor prognosis and important for personalized medicine. Eighty-one HCC specimens were used in this study. We screened for IHR-related genes by DNA microarray analysis. The validation of screening was performed by using real-time PCR. The methylation levels in genomic DNAs were measured by quantitative methylation-specific PCR. Six hepatoma cell lines were used for examination of ABCB6 expressional regulation. Time-to-event analyses for recurrence after surgery were analyzed by Kaplan-Meier analysis and Cox regression analysis with cutoff values obtained from receiver operating characteristic (ROC) analysis. We confirmed that ABCB6 mRNA levels were significantly higher in hepatitis C virus (HCV)-related HCCs with early IHR compared to HCV-related HCCs without early IHR (2.5-fold, P=0.01) and the corresponding non-cancerous livers (3.1-fold, P=0.05). Experiments with cell lines showed correlation between DNA methylation and mRNA levels of ABCB6. ROC analysis revealed that mRNA levels (0.81 area under the curve, 88% sensitivity and 72% specificity) and DNA methylation levels (0.81 area under the curve, 80% sensitivity and 80% specificity) of ABCB6 in HCV-related HCCs allowed for the accurate discrimination of the development of early IHR. Cox regression analysis revealed that ABCB6 mRNA levels was an independent risk factor for IHR of HCV-related HCC. Aberrant mRNA and DNA methylation levels of ABCB6 may serve as useful predictive biomarkers for early IHR of HCV-related HCC.


[Effects of rifampicin and streptomycin on the concentrations of testosterone and cortisol in the blood of men (author's transl)].
It has been shown previously that the biosynthesis and metabolism of oestrogens is affected by rifampicin. The present study was undertaken to investigate if rifampicin may also influence the plasma concentrations of testosterone and cortisol. Five male patients suffering from tuberculosis received rifampicin or streptomycin. During treatment with rifampicin testosterone concentrations were significantly elevated as compared to control conditions; this increase was found in the morning (8.00 h) as well as in the afternoon (16.30 h). When the same patients were treated with streptomycin, only irregular changes of testosterone concentrations were observed. The blood level of cortisol was increased in two patients, but unchanged in three patients who were treated with rifampicin. No changes at all were detected in patients under treatment with streptomycin. The results presented here show that rifampicin -- in contrast to streptomycin -- influences the biosynthesis of testosterone and possibly also that of cortisol.


Targeting of adenovirus via genetic modification of the viral capsid combined with a protein bridge.
A potential barrier to the development of genetically targeted adenovirus (Ad) vectors for cell-specific delivery of gene therapeutics lies in the fact that several types of targeting protein ligands require posttranslational modifications, such as the formation of disulfide bonds, which are not available to Ad capsid proteins due to their nuclear localization during assembly of the virion. To overcome this problem, we developed a new targeting strategy, which combines genetic modifications of the Ad capsid with a protein bridge approach, resulting in a vector-ligand targeting complex. The components of the complex associate by virtue of genetic modifications to both the Ad capsid and the targeting ligand. One component of this mechanism of association, the Fc-binding domain of Staphylococcus aureus protein A, is genetically incorporated into the Ad fiber protein. The ligand is comprised of a targeting component fused with the Fc domain of immunoglobulin, which serves as a docking moiety to bind to these genetically modified fibers during the formation of the Ad-ligand complex. The modular design of the ligand solves the problem of structural and biosynthetic compatibility with the Ad and thus facilitates targeting of the vector to a variety of cellular receptors. Our study shows that targeting ligands incorporating the Fc domain and either an anti-CD40 single-chain antibody or CD40L form stable complexes with protein A-modified Ad vectors, resulting in significant augmentation of gene delivery to CD40-positive target cells. Since this gene transfer is independent of the expression of the native Ad5 receptor by the target cells, this strategy results in the derivation of truly targeted Ad vectors suitable for tissue-specific gene therapy.


Risk of colorectal polyps and of malignancies in asymptomatic carriers of mutations in the main DNA mismatch repair genes.
Mutation carriers (Mut+) in DNA mismatch repair genes are predisposed to cancer of various organs and to adenomatous polyps; however, they may remain asymptomatic and cancer or polyp-free for several years. We purposed to analyse the clinical follow-up of individuals carrying constitutional mutations in the MLH1, MSH2 or MSH6 genes who were unaffected by benign polyps or malignant tumours at diagnosis. Mut + subjects (n.81) were members of Lynch syndromes in whom mutations were detected between 1993 and 2015; all were asymptomatic at diagnosis. They were informed of the cancer risk and surveillance was suggested. As controls, 113 nongene carriers (Mut-) in the same Lynch families were identified. About one-fourth of the mutation carriers developed polyps, mostly adenomas; polyps were less (12%, p < .05) in Mut - subjects, and hyperplastic lesions were the prevalent histology. More polyps were detected in MLH1 vs. MSH2 mutation carriers. In Mut+, 21 malignant tumours developed in 14 carriers vs. 4 tumours in 3 patients among Mut- (p < .001). Tumours were mostly of the Lynch spectrum; however, three glioblastomas were developed, together with neoplasms of various organs (duodenum, thyroid, skin, lung and cervix). Mean age of tumour occurrence was 43.0 years in Mut + vs. 53.0 among Mut-. Cancer developed more often in Mut+, with no consistent difference between MLH1 and MSH2 carriers. More polyps (mostly adenomas) were detected in MLH1 carriers. The majority (13 of 21) of malignant tumours occurred in organs for which there is no recommended surveillance, and were lethal in three patients.


The architecture of mammalian ribosomal protein promoters.
Mammalian ribosomes contain 79 different proteins encoded by widely scattered single copy genes. Coordinate expression of these genes at transcriptional and post-transcriptional levels is required to ensure a roughly equimolar accumulation of ribosomal proteins. To date, detailed studies of only a very few ribosomal protein (rp) promoters have been made. To elucidate the general features of rp promoter architecture, I made a detailed sequence comparison of the promoter regions of the entire set of orthologous human and mouse rp genes. A striking evolutionarily conserved feature of most rp genes is the separation by an intron of the sequences involved in transcriptional and translational regulation from the sequences with protein encoding function. Another conserved feature is the polypyrimidine initiator, which conforms to the consensus (Y)2C+1TY(T)2(Y)3. At least 60 % of the rp promoters contain a largely conserved TATA box or A/T-rich motif, which should theoretically have TBP-binding capability. A remarkably high proportion of the promoters contain conserved binding sites for transcription factors that were previously implicated in rp gene expression, namely upstream GABP and Sp1 sites and downstream YY1 sites. Over 80 % of human and mouse rp genes contain a transposable element residue within 900 bp of 5' flanking sequence; very little sequence identity between human and mouse orthologues was evident more than 200 bp upstream of the transcriptional start point. This analysis has provided some valuable insights into the general architecture of mammalian rp promoters and has identified parameters that might coordinately regulate the transcriptional activity of certain subsets of rp genes.


Participation of internal medicine department chairs in the internal medicine clerkship--results of a national survey.
To characterize the involvement of internal medicine department chairs in the core third-year internal medicine clinical clerkship. In 2003, the Clerkship Directors in Internal Medicine (CDIM) surveyed its membership. Along with demographics, clerkship directors were asked if the department's chair participated in the clerkship, the number of hours per month the clerkship director and chair discussed clerkship issues, and if published job expectations were discussed. The response rate was 62% (158/254): 103 responses (89 clerkship directors) represented unique medical schools, which formed the basis of the analysis. Eighty-two percent (84/103) reported the chair taught in the clerkship: 54% as teaching attending, 53% as ward attending, 13% as ambulatory attending, and 20% other (e.g., lectures, student rounds, morning report). Of them, 36% performed two activities; 14% three activities; and 2% four activities. Thirty-six percent of the clerkship directors discussed published expectations with their chair. They spent 1.7 (SD 2.2) hours per month with the chair discussing clerkship issues. However, 17% spent zero hours per month with the chair, and 29% spent zero hours per month with a dean's office representative. Chairs who taught spent more time each month with the clerkship director compared with chairs who did not teach (1.9 versus .82 hours, p = .01, Mann-Whitney). There was no association between the chairs' teaching and clerkship directors' demographics. Internal medicine department chairs are significantly involved in the clinical education of medical students, both administratively and through direct teaching. Chairs who teach spend more time discussing clerkship issues with the clerkship director. Chairs and clerkship directors should discuss expectations, and chairs should continue to visibly demonstrate their commitment to students' education.


Quantitative assessment of P-glycoprotein function in the rat blood-brain barrier by distribution volume of [11C]verapamil measured with PET.
The blood-brain barrier (BBB) is a functional barrier that hampers the delivery of various drugs to the brain by its physicoanatomical properties and by the presence of ATP-driven drug efflux pumps, such as P-glycoprotein (P-gp). The aims of this study were (1) to study whether the distribution volume (DV) is useful for quantification of (labeled) P-gp substrate kinetics over the BBB and (2) to study how brain DV is affected by P-gp modulation. We measured the kinetics of the P-gp substrate [11C]verapamil (0.1 mg/kg) in rat brains using positron emission tomography (PET) and arterial blood sampling. Cyclosporin A (CsA) at 0, 10, 15, 25, 35, and 50 mg/kg of body weight was used as a P-gp modulator. The [11C]verapamil kinetics were very well described by DV, computed by noncompartmental Logan analysis. Logan analysis resulted in excellent fits of dynamic PET data, revealing the reversible behavior of [11C]verapamil and its associated DV. The DV in unmodulated rats was 0.65 ml/ml +/- 0.23 (mean +/- SD). After modulation with 10, 15, 25, 35, and 50 mg/kg of CsA, DV values increased to 0.82 +/- 0.06, 1.04 +/- 0.20, 2.85 +/- 0.51, 2.91 +/- 0.64, and 3.77 +/- 1.23, respectively. The [11C]Verapamil kinetics were saturable at modulation levels above 25 mg/kg of CsA. The data fitted well by a four-parameter Hill plot (R2 = 0.79). In conclusion, the DV of [11C]verapamil is a valid and potent tool to measure the kinetics of (labeled) P-gp substrates in vivo at the BBB. The brain DV of [11C]verapamil increases dose dependently by P-gp modulation. Quantitative insight into in vivo P-gp modulation may be a promising step toward assessment of P-gp substrate delivery to human brains.


PSGL-1, a Strategic Biomarker for Pathological Conditions in HIV Infection: A Hypothesis Review.
P-selectin glycoprotein ligand-1 (PSGL-1) has been established to be a cell adhesion molecule that is involved in the cellular rolling mechanism and the extravasation cascade, enabling the recruitment of immune cells to sites of inflammation. In recent years, researchers have established that PSGL-1 also functions as an HIV restriction factor. PSGL-1 has been shown to inhibit the HIV reverse transcription process and inhibit the infectivity of HIV virions produced by cells expressing PSGL-1. Cumulative evidence gleaned from contemporary literature suggests that PSGL-1 expression negatively affects the functions of immune cells, particularly T-cells, which are critical participants in the defense against HIV infection. Indeed, some researchers have observed that PSGL-1 expression and signaling provokes T-cell exhaustion. Additionally, it has been established that PSGL-1 may also mediate virus capture and subsequent transfer to permissive cells. We therefore believe that, in addition to its beneficial roles, such as its function as a proinflammatory molecule and an HIV restriction factor, PSGL-1 expression during HIV infection may be disadvantageous and may potentially predict HIV disease progression. In this hypothesis review, we provide substantial discussions with respect to the possibility of using PSGL-1 to predict the potential development of particular pathological conditions commonly seen during HIV infection. Specifically, we speculate that PSGL-1 may possibly be a reliable biomarker for immunological status, inflammation/translocation, cell exhaustion, and the development of HIV-related cancers. Future investigations directed towards our hypotheses may help to evolve innovative strategies for the monitoring and/or treatment of HIV-infected individuals.


[Myocardial serotonin metabolism after local ischemia and ischemic precondition].
To determine the effect of ischemic preconditioning upon myocardial serotonin and 5-hydroxyindolacetic acid (5-HIAA) dynamic in myocardial ischemia and reperfusion. 28 male Wistar rats anesthetized with urethane were randomly divided into 2 groups. In the control group (n = 13) rats were subjected to 30 min coronary occlusion and subsequent 120 min reperfusion. In the ex- perimental group (n = 15) ischemic preconditioning (3 x 3 min ischemia + 3 x 3 min reperfusion) before prolonged ischemia was used. Myocardial interstitial serotonin and 5-HIAA were measured using a microdialysis technique. Myocardial serotonin and 5-HIAA significantly increased af- ter ischemic preconditioning (p = 0.00298; p = 0.00187). In prolonged ischemia interstitial serotonin level was lower in the experimental group vs. control up to 20 min of ischemia (p < 0.05). We conclude that ischemic preconditioning increases interstitial myocardial serotonin, but inhibit serotonin increase in subsequent prolonged myocardial ischemia. After 20 minutes of reperfusion the lack of correlation between serotonin and 5-HIAA levels appeared which may be the evidence of serotonin uptake activation.


Evaluation of Zn, Cu, and Se Levels in the North American Autism Spectrum Disorder Population.
Metal ion dyshomeostasis and disparate levels of biometals like zinc (Zn), copper (Cu), and selenium (Se) have been implicated as a potential causative factor for Autism Spectrum Disorder (ASD). In this study, we have enrolled 129 children (aged 2-4 years) in North America, of which 64 children had a diagnosis of ASD and 65 were controls. Hair, nail, and blood samples were collected and quantitatively analyzed for Zn, Cu and Se using inductively coupled plasma mass spectrometry (ICP-MS). Of the analyzed biometals, serum Se (116.83 ± 14.84 mcg/mL) was found to be significantly lower in male ASD cases compared to male healthy controls (128.21 ± 9.11 mcg/mL; p < 0.005). A similar trend was found for nail Se levels in ASD (1.01 ± 0.15 mcg/mL) versus that of controls (1.11 ± 0.17 mcg/mL) with a p-value of 0.0132 using a stratified Wilcoxon rank sum testing. The level of Se in ASD cohort was co-analyzed for psychometric correlation and found a negative correlation between total ADOS score and serum Se levels. However, we did not observe any significant difference in Zn, Cu, and Zn/Cu ratio in ASD cases versus controls in this cohort of North American children. Further studies are recommended to better understand the biology of the relationship between Se and ASD status.


Collagen fibril bundles: a branching assembly unit in tendon morphogenesis.
The assembly, deposition and organization of collagen fibril bundles and their composite fibrils were studied during morphogenesis of the chick embryo tendon using electron microscopy, serial sections and computer-assisted three-dimensional reconstruction techniques. The 14-day chick embryo is a stage when tendon architecture is being established and rapid changes in the mechanical properties occur between days 14 and 17 of development. Tendon matrix structure develops from discrete subunits, bundles of collagen fibrils. The bundles branch; undergo a gradual rotation over several micrometers; are intimately associated with the cellular elements of the developing tendon; and form arborizing networks within and among fascicles. The organization of discrete fibril segments into bundles, during the establishment of tendon architecture and function, where the segmental fibrillar components could interact with the interfibrillar matrix as well as with adjacent fibrils would contribute to the stabilization of this structure. The observed gradual rotation of the bundles would serve to stabilize the immature bundle through the physical twining of the composite fibrils while the extensive branching of the bundles observed at 14-days of development and their intimate association with the cellular elements would provide a higher order of structure stabilization.


The mental model theory of spatial reasoning re-examined: the role of relevance in premise order.
The effect of premise order on spatial reasoning was investigated using a paradigm introduced by Byrne & Johnson-Laird in 1989. Three kinds of problems were developed: the first of these was expected to prompt one mental model, whereas the second and third were expected to prompt two mental models. The two-model problems differed from each other by the position (first or last) of a premise which was responsible for an indeterminacy. On the basis of Sperber & Wilson's Relevance theory, published in 1986 and 1995, it was predicted and observed that a two-model problem with the crucial premise in the last position was easier than a two-model problem with the crucial premise in the first position. Moreover, it was predicted and observed that the two-model problem with the crucial premise in the last position was as easy as the one-model problem. Implications of these results for the standard mental model theory of spatial reasoning are discussed.


Primary open-angle glaucoma.
Glaucoma is an optic neuropathy that is characterized by the progressive degeneration of the optic nerve, leading to visual impairment. Glaucoma is the main cause of irreversible blindness worldwide, but typically remains asymptomatic until very severe. Open-angle glaucoma comprises the majority of cases in the United States and western Europe, of which, primary open-angle glaucoma (POAG) is the most common type. By contrast, in China and other Asian countries, angle-closure glaucoma is highly prevalent. These two types of glaucoma are characterized based on the anatomic configuration of the aqueous humour outflow pathway. The pathophysiology of POAG is not well understood, but it is an optic neuropathy that is thought to be associated with intraocular pressure (IOP)-related damage to the optic nerve head and resultant loss of retinal ganglion cells (RGCs). POAG is generally diagnosed during routine eye examination, which includes fundoscopic evaluation and visual field assessment (using perimetry). An increase in IOP, measured by tonometry, is not essential for diagnosis. Management of POAG includes topical drug therapies and surgery to reduce IOP, although new therapies targeting neuroprotection of RGCs and axonal regeneration are under development.


Long-read RNA-seq atlas of novel microglia isoforms elucidates disease-associated genetic regulation of splicing.
Microglia, the innate immune cells of the central nervous system, have been genetically implicated in multiple neurodegenerative diseases. We previously mapped the genetic regulation of gene expression and mRNA splicing in human microglia, identifying several loci where common genetic variants in microglia-specific regulatory elements explain disease risk loci identified by GWAS. However, identifying genetic effects on splicing has been challenging due to the use of short sequencing reads to identify causal isoforms. Here we present the isoform-centric microglia genomic atlas (isoMiGA) which leverages the power of long-read RNA-seq to identify 35,879 novel microglia isoforms. We show that the novel microglia isoforms are involved in stimulation response and brain region specificity. We then quantified the expression of both known and novel isoforms in a multi-ethnic meta-analysis of 555 human microglia short-read RNA-seq samples from 391 donors, the largest to date, and found associations with genetic risk loci in Alzheimer's disease and Parkinson's disease. We nominate several loci that may act through complex changes in isoform and splice site usage.


Identification and expression analysis of zebrafish testis-specific gene 10 (tsga10).
Several clinical studies suggest that testis-specific gene antigen 10 (TSGA10) is a cancer-testis antigen with a discernible expression pattern in the testis. Recent studies have highlighted that TSGA10 overexpression in HeLa cells impairs the transcriptional activity of hypoxia-inducible factor alpha (HIF-1α) and inhibits angiogenesis. In this study, we used the zebrafish as a powerful model organism to identify and characterize the orthologue of TSGA10. We analyzed the gene expression pattern by RT-PCR and whole mount in situ hybridization and overexpressed the tsga10 protein by mRNA microinjection. Our results revealed that during early development, tsga10 expression is enriched, but gradually subsides between 0 and 72 hours post fertilization (hpf). There was no detectable transcript at the larval stages. In adult fish, we found high expression levels of tsga10 in the testis and unfertilized egg and low levels of gene expression in the brain, eyes and muscle. Overexpression of tsga10, using tsga10 mRNA microinjection into one-cell stage embryos, resulted in angiogenic and morphological defects at 24 and 48 hpf. This study clarified the expression pattern of tsga10 in different developmental stages and adult tissues, suggesting that tsga10 may have a related biological role in different cell types and tissues. Our results indicate that tsga10 mRNA at embryonic stages is maternally deposited, indicating a transient functional role during embryogenesis. Our findings suggest that tsga10 is a human orthologous gene relevant for future studies to elucidate its mechanism of action in angiogenesis.


A modified method for the in vivo labeling of red blood cells with Tc-99m: concise communication.
The rate of incorporation of Tc-99m into red blood cells pretinned in vivo was measured by collecting blood samples in stannous DTPA solution, which served as a competing ligand for Tc-99m. This collection technique permitted a measurement of high-affinity red-cell labeling efficiency at the instant of sampling. At 0.5 min after injection only 62% of technetium is tightly bound to the red cell; this rises to 94.5% at 10 min. Based on the graded labeling of the red cells, the in vivo labeling procedure was modified by isolating pertechnetate and red blood cells tinned in vivo in a syringe during the first 10 min of labeling. The pertechnetate is thus prevented from distributing to extravascular compartments, and 90% of the injected Tc-99m is firmly bound to red blood cells at the time of injection. In a series of 23 patients, seven were tested with the in vivo method and seven with the modified in vivo method, and nine patients were tested with each method on separate occasions. A decrease in gastric activity and improved image quality were found with the modified method compared with the standard method of in vivo red-cell labeling.


Clinical features affecting efficacy of immune checkpoint inhibitors in pretreated patients with advanced NSCLC: a Danish nationwide real-world study.
Immune checkpoint inhibitors (ICIs) are implemented as standard treatment for patients with advanced non-small cell lung cancer (NSCLC) in first-line and subsequent-line treatment. However, certain subgroups such as patients with older age, poor performance status (PS), and severe comorbidity are underrepresented in the randomized controlled trials (RCTs). This study aimed to assess overall survival (OS), treatment data, and clinical features affecting second- or subsequent-line ICI efficacy in an unselected, Danish, nationwide NSCLC population. Patients with advanced NSCLC who started nivolumab or pembrolizumab as second-line or subsequent-line treatment between 1 September 2015, and 1 October 2018, were identified from institutional records of all Danish oncology departments. Clinical and treatment data were retrospectively collected. Descriptive statistics and survival analyses were performed. Data were available for 840 patients; 49% females. The median age was 68 years (19% were ≥75 years), 19% had PS ≥2, and 36% had moderate to severe comorbidity. The median OS (mOS) was 12.2 months; 15.1 months and 10.0 months in females and males, respectively. The median time-to-treatment discontinuation (mTTD) and median progression-free survival (mPFS) was 3.2 and 5.2 months, respectively. Patients with PS ≥2 had a mOS of 4.5 months, mTTD of 1.1 month, and mPFS of 2.0 months. In multivariable Cox regression analysis, male sex (HR = 1.35, 95% CI 1.11-1.62), PS >0 (PS 1, HR = 1.88, 95% CI 1.52-2.33; PS ≥2, HR = 4.15, 95% CI 3.13-5.5), liver metastases (HR = 1.72, 95% CI 1.34-2.22), and bone metastases (HR = 1.27, 95% CI 1.03-1.58) were significant poor prognostic OS factors. Danish real-world patients with advanced NSCLC treated with second- or subsequent-line ICI had an OS comparable to results from RCTs. Women, frail and older patients constituted a higher proportion than in previous RCTs. Clinical features associated with poor OS were male sex, PS ≥1 (in particular PS ≥2), bone-, and liver metastases.


Immunocytochemical assay for estrogen receptor: relationship to outcome of therapy in patients with advanced breast cancer.
We have used an immunoperoxidase technique utilizing a monoclonal antibody to the estradiol receptor to identify immunoreactive estradiol receptor in breast carcinomas and have examined the relationship between the immunoreactive estradiol receptor and response to therapy in patients with advanced breast cancer. Fifty-six patients were found to be assessable for response to endocrine therapy. Twenty-two showed an objective response to some form of endocrine manipulation, and all these had positively stained carcinomas. None of the 17 patients with negatively stained carcinomas responded to endocrine therapy. We conclude that the monoclonal antibody to estradiol receptor can help identify breast cancer patients who may respond to endocrine therapy.


Ethics review: 'Living wills' and intensive care--an overview of the American experience.
Withdrawal and limitation of life support in the intensive care unit is common, although how this decision is reached can be varied and arbitrary. Inevitably, the patient is unable to participate in this discussion because their capacity is limited by the nature of the illness and the effects of its treatment. Physicians often discuss these decisions with relatives in an attempt to respect the patient's wishes despite evidence suggesting that the relatives may not correctly reflect the patient's desires. Advance decisions, commonly known as 'living wills', have been proposed as a way of facilitating the maintenance of an individual's autonomy when they become incapacitated. Others have argued that legalising advance decisions is euthanasia by the back door. In October 2007 in England and Wales, advance decisions will become legally binding as part of the 2005 Mental Capacity Act. This has been the case in the USA for many years. The purpose of the present review is to examine the published literature regarding the effect of advance decisions in relation to the provision of adult critical care.


Rheumatoid arthritis: the role of the kinder and gentler therapies.
Rehabilitative therapies, as adjuncts to pharmacological and surgical therapies in rheumatoid arthritis (RA), help minimize pain and inflammation, improve functional capabilities, and, above all, enhance quality of life. The effects of heat, cold, and electrical stimulation on neurohumoral, inflammatory, and immunological mechanisms has been noted, although knowledge about their mode of action is lacking. There is, however, much that these modalities, in addition to splints, bracing, exercise, activities-of-daily-living training, and psychological, social, and vocational counseling can offer to reduce suffering and disability in patients with RA. Far more, however, needs to be learned to provide precise prescriptions and more effective applications.


A guinea-pig model of ultrasonically nebulized distilled water-induced bronchoconstriction.
Ultrasonically nebulized distilled water-induced bronchoconstriction (UNDW-IB) is specific to asthma. The mechanisms underlying UNDW-IB are not fully understood, and no reproducible animal model has been reported. The purpose of this study was to develop a guinea-pig model of UNDW-IB. Ultrasonically nebulized distilled water (UNDW) was inhaled 20 min after an aerosolized antigen challenge in passively sensitized and artificially ventilated guinea-pigs. UNDW was also inhaled 5 and 20 min after 0.1 mg x mL(-1) methacholine inhalation in nonsensitized animals. In addition, 0.1 mg x kg(-1) S-1452, a thromboxane A2 antagonist, or saline was given intravenously 5 min before UNDW inhalation in sensitized animals. The inhalation of UNDW caused bronchoconstriction, when inhaled 20 min after an antigen challenge in sensitized guinea-pigs. UNDW inhalation did not produce bronchoconstriction after saline inhalation in nonsensitized or sensitized guinea-pigs, or after antigen inhalation in nonsensitized animals. Methacholine-induced bronchoconstriction did not evoke UNDW-IB. Neither did S-1452 reduce the UNDW-IB. In conclusion, the guinea-pig model of ultrasonically nebulized distilled water-induced bronchoconstriction developed in this study suggests that allergic reaction, but not bronchoconstriction, can induce bronchial hyperresponsiveness to ultrasonically nebulized distilled water, and that thromboxane A2 is not involved in ultrasonically nebulized distilled water-induced bronchoconstriction.


Monoclonal anti-envelope antibody AP33 protects humanized mice against a patient-derived hepatitis C virus challenge.
End-stage liver disease (ESLD) caused by hepatitis C virus (HCV) infection is a major indication for liver transplantation. However, immediately after transplantation, the liver graft of viremic patients universally becomes infected by circulating virus, resulting in accelerated liver disease progression. Currently available direct-acting antiviral therapies have reduced efficacy in patients with ESLD and prophylactic strategies to prevent HCV recurrence are still highly needed. In this study, we compared the ability of two broadly reactive monoclonal antibodies (mAbs), designated 3/11 and AP33, recognizing a distinct, but overlapping, epitope in the viral E2 glycoprotein to protect humanized mice from a patient-derived HCV challenge. Their neutralizing activity was assessed using the HCV pseudoparticles and cell-culture-derived HCV systems expressing multiple patient-derived envelopes and a human-liver chimeric mouse model. HCV RNA was readily detected in all control mice challenged with a patient-derived HCV genotype 1b isolate, whereas 3 of 4 AP33-treated mice were completely protected. In contrast, only one of four 3/11-treated mice remained HCV-RNA negative throughout the observation period, whereas the other 3 had a viral load that was indistinguishable from that in the control group. The increased in vivo efficacy of AP33 was in line with its higher affinity and neutralizing capacity observed in vitro. Although mAbs AP33 and 3/11 target the same region in E2, only mAb AP33 can efficiently protect from challenge with a heterologous HCV population in vivo. Given that mAb AP33 efficiently neutralizes viral variants that escaped the humoral immune response and reinfected the liver graft of transplant patients, it may be a valuable candidate to prevent HCV recurrence. In addition, our data are valuable for the design of a prophylactic vaccine.


A human liver cell-based system modeling a clinical prognostic liver signature for therapeutic discovery.
Chronic liver disease and hepatocellular carcinoma (HCC) are life-threatening diseases with limited treatment options. The lack of clinically relevant/tractable experimental models hampers therapeutic discovery. Here, we develop a simple and robust human liver cell-based system modeling a clinical prognostic liver signature (PLS) predicting long-term liver disease progression toward HCC. Using the PLS as a readout, followed by validation in nonalcoholic steatohepatitis/fibrosis/HCC animal models and patient-derived liver spheroids, we identify nizatidine, a histamine receptor H2 (HRH2) blocker, for treatment of advanced liver disease and HCC chemoprevention. Moreover, perturbation studies combined with single cell RNA-Seq analyses of patient liver tissues uncover hepatocytes and HRH2+, CLEC5Ahigh, MARCOlow liver macrophages as potential nizatidine targets. The PLS model combined with single cell RNA-Seq of patient tissues enables discovery of urgently needed targets and therapeutics for treatment of advanced liver disease and cancer prevention.


Wireless real-time electronic data capture for self-assessment of motor function and quality of life in Parkinson's disease.
Frequent assessment of the symptoms in Parkinson's disease (PD) is important in both clinical and experimental settings, especially when motor fluctuations are present. Patient diaries are increasingly used in studies, allowing patients to stay in their home environments. However, traditional paper diaries may not reflect reality because of a lack in compliance or retrospective data entries. This study presents a comparison between paper diaries and a new method, real-time data capture with a hand-held computer (electronic diary). Twenty patients with PD diagnosed at least 5 years previously were randomly assigned to use either a paper diary or an electronic diary on 8 days during 1 month. Questions were answered every 2 hours over a 12-hour period on each day. Median compliance was 88% with the electronic diary and 98% with the paper diary, although strict compliance to the scheduled times by patients using the paper diary was 78%. Neither age nor earlier experience with computers affected the patient's ability to use the electronic diary. Electronic diaries can be used for self-assessment of PD symptoms. The real-time feature provides fast access to clean data with knowledge of true compliance.


A rare case of hepatic T-cell rich B-cell lymphoma (TCRBCL) in a juvenile dog.
A 7-month-old castrated male French Bull dog was presented with vomiting, lethargy, anorexia and weight loss of 2 weeks duration. The patient's history and clinical manifestations of suspected hepatopathy were subjected to ultrasonography, radiography, biochemical investigations and cytology of hepatic lesion. The cytologic impression was hepatic lymphoma, which was later confirmed by histopathology. The neoplastic cells were strongly diffusely immunoreactive for PAX5, but not immunoreactive for CD3, and B lymphocyte specific clonal proliferation was detected using by assay of antigen receptor rearrangement. Large numbers of immunoreactive mature non-neoplastic lymphocytes were admixed with the neoplastic cell population. Therefore, the immunohistochemical results were definitively consistent with a T-cell rich B-cell lymphoma (TCRBCL). This is the first description of a hepatic TCRBCL in a juvenile dog showing a poor response to aggressive chemotherapy.


Development of a bioassay for ovarian carcinoma colony-forming cells.
We have reviewed the application of our in vitro assay for human tumor stem cells to the cloning of human ovarian adenocarcinoma cells in soft agar. Tumor colonies grew from both effusions and biopsies from 85% of more than 100 ovarian cancer patients tested. Up to 2,000 colonies appeared after 10 to 14 days in culture, yielding a maximum plating efficiency of 1%. Cells from nonmalignant effusions did not form colonies under these conditions. The number of tumor colonies was proportional to the number of cells plated between concentrations of 104 to 106 cells/dish. Morphological and histochemical criteria showed that the colonies consisted of cells with the same characteristics as those of the original tumor. H3Tdr suicide colony-forming cells were actively in transient through the cell cycle. Removal of phagocytic cells with carbonyl iron markedly reduced the plating efficiency, and 2-mercaptoethanol could only partially substitute for macrophages. Spleen cell-conditioned medium from oil-primed BALB/c mice was not required. Endogenous macrophages within the tumor may provide the conditioning factor or factors required for in vitro growth. Thus, this assay is proving extremely useful for studying the biology and drug sensitivity of human ovarian cancer.


Tuberculosis Arthritis in the Wrist While Using Rituximab for Rheumatoid Arthritis Treatment.
Data from clinical trials indicate that there are no increased risk of tuberculosis (TB) infections in rheumatoid arthritis (RA) patients while using rituximab (RTX). Herein, we report a RA patient who developed TB arthritis while using RTX. A 49-year-old patient was treated with methotrexate and prednisolone along with RTX for two years. Later, she presented with increasing pain, swelling, redness and cutaneous fistulization in her left wrist for two months. The lesion on the wrist was debritted. Histopathologic evaluation revealed the presence of acid-fast bacilli. Polymerase chain reaction test and culture confirmed mycobacterium tuberculosis. RTX, methotrexate and prednisolone were withdrawn. The patient was treated with 12-month course of antituberculous treatment and responded well. The patient, who did not have pain or swelling in her other joints, was not given any treatment for RA after antituberculous treatment. Clinicians should keep in mind that TB infections may be encountered while using RTX. Latent TB screening may be appropriate in patients using concomitant corticosteroid and living in TB endemic areas.


Update on the epidemiology and management of cryptococcal meningitis.
Despite recent improvements in the diagnosis and treatment of cryptococcosis, cryptococcal meningitis is responsible for > 600,000 deaths/year worldwide. The aim of this work is to provide an update on the developments in its epidemiology and management. Understanding the pathogenesis of Cryptococcus has improved, and new insights for the virulence of the fungus and the host response have enabled scientists to design new ways to confront this infection. Additionally, invertebrate model hosts have greatly facilitated the research in this field. Importantly, the epidemiology of Cryptococcus gattii has continued to evolve, and the emergence of this highly virulent species in immunocompetent populations, especially in Northwestern America and British Columbia, warrants increased awareness because delayed diagnosis and inappropriate antifungal therapy is associated with high mortality. Diagnosis remains a challenge, but new techniques for early and inexpensive identification of the pathogen are under development. Management can vary, based on the patient population (HIV-seropositive, organ transplant recipients or non-transplant/non-HIV). In most patients, amphotericin B with flucytosine continues to be the most appropriate induction therapy. However, in organ transplant recipients the use of liposomal amphotericin B improves mortality compared with deoxycholate amphotericin B. Also, the combination of amphotericin B with fluconazole seems to be a reasonable alternative, while fluconazole with flucytosine is superior to fluconazole monotherapy.


[Molluscum contagiosum disclosing HTLV III infection].
We report here a case of cutaneous molluscum contagiosum infection, which led to the diagnosis of HTL V III infection. Profuse mollusca contagiosa are one of the cutaneous lesions indicating the presence of AIDS at any stage.


Intranasal administration of milnacipran in rats: evaluation of the transport of drugs to the systemic circulation and central nervous system and the pharmacological effect.
Recently, transnasal drug delivery has attracted a great deal of attention as an administration route to deliver drugs directly to the central nervous systems (CNS) and drug targeting of the CNS is expected to increase. In the present study, we investigated the possibility of using a transnasal delivery system for milnacipran, a serotonin-noradrenaline reuptake inhibitor (SNRI), by evaluating the transport to the systemic circulation and cerebrospinal fluid (CSF) and the pharmacological effect after intranasal (i.n.) administration. Moreover, the effect of chitosan as a bioadhesive material on the transport to the systemic circulation and CSF and the pharmacological effect after i.n. administration were evaluated. As a result, i.n. administration of milnacipran was found to produce a higher direct delivery to the CNS as well as to the systemic circulation, suggesting that this is a promising route of administration and an alternative to peroral (p.o.) administration. Furthermore, the i.n. co-administration with chitosan led to increased plasma and CSF concentrations and an enhanced pharmacological effect, evaluated by means of the forced swimming test. The results suggested that chitosan produced a long residence time of milnacipran in the nasal cavity due to its bioadhesive effect, leading to the enhanced transport of milnacipran from the systemic circulation to the CNS via the blood-brain barrier by an increase in systemic absorption as well as direct transport to the CNS, resulting in a higher antidepressant effect compared to that with p.o. administration.


Unjustified variations in patient management for acute myocardial infarction in the Rhône-Alpes region of France. PRIMA Group.
To examine determinants of use of cardiac procedures after acute myocardial infarction and identify variation factors. Observational prospective cohort study of 2,519 patients in 48 centers with a two-level logistic-regression analysis. Elderly patients were less likely to undergo pre- and inhospital thrombolysis (odds ratios, 0.71 and 0.64; 95% CI, 0.62-0.81 and 0.58-0.69, respectively). The elderly, females, and patients with heart failure on admission were less likely to undergo noninvasive tests (0.74, 0.62, and 0.51; 95% CI, 0.67-0.81, 0.46-0.83, and 0.38-0.68, respectively) and coronary angiography (0.38, 0.53, and 0.67; 95% CI, 0.34-0.42, 0.38-0.74, and 0.52-0.86, respectively) but not revascularization. Hospital factors were more difficult to interpret. Elderly, women, and heart failure patients underwent fewer cardiac procedures than lower-risk patients. Physicians should change their attitude toward these groups and use advanced procedures, bearing in mind the patients' needs rather than good procedural outcomes.


Microtechnique for serum opacity factor characterization of group A streptococci adaptable to the use of human sera.
We have developed a microtechnique for detection of streptococcal serum opacity factor (OF) and for typing of group A streptococci by inhibition of OF. This technique, which involves the use of single wells of standard 96-well tissue culture plates, offers several advantages over previous methods: no advance test preparation is required, allowing tests to be quickly and easily performed; only small quantities of reagents are required; results can be determined visually (qualitative) or by using a photometric enzyme-linked immunosorbent assay plate reader (quantitative); and human serum samples may be quickly and easily screened for OF-inhibitory antibody and subsequently used in place of difficult-to-produce and expensive hyperimmune animal sera for OF characterization of group A streptococci. Fifty-eight samples of normal adult human serum were tested by this new microtechnique for anti-OF antibodies, and 49 (84%) were found to have antibody against 1 or more of the 27 recognized OF-positive serotypes. OF antibodies to M-4, M-2, M-75, and M-48 were most common in these individuals. These 58 human serum samples collectively contained antibody to 25 of the 27 different OF-producing serotypes. Serum samples from four individuals were tested for persistence of OF antibody. OF antibodies to eight different serotypes present in the serum samples collected 7 to 12 years previously were present in the freshly collected sera, indicating that OF antibody persists in human antisera for many years. This new technique has distinct advantages and makes it possible for many laboratories to use this technique to characterize group A streptococci.


[Folate and Pregnancy, current concepts: It is required folic acid supplementation?]
Folate intake during pregnancy is essential for an adequate fetal and placental development and for the long time health of the individual. Its deficiency may induce fetal pathologies, including neural tube disease (NTD). Therefore, several countries implemented public policies to fortify foods with folic acid (FA). Chile started the fortification of wheat flour with FA in the year 2000, decreasing a 43% the prevalence of NTD. However, despite the high consumption of bread (the main fortified food with FA) by our population, a high number of pregnant women consume FA supplements, thus, over passing the maximal recommended FA intake. Additionally, if the diet is reduced in vitamin B12, the optimal ratio folates/vit B12 may be altered, thus inducing changes in the methylation of specific genes and other metabolic pathways, affecting fetal development and the long-term health of the neonates. We think that, after 16 years of the initiation of the fortification of wheat flour with FA, it is necessary to evaluate the possible side effects of a high intake of FA in the pregnant population and their offspring. This article shows antecedents about mechanisms of folates and vit B12 at cellular level, and their possible consequences of an elevated FA maternal intake on the offspring.


Kinetics of capillary condensation in nanoscopic sliding friction.
The velocity and humidity dependence of nanoscopic sliding friction has been studied on CrN and diamondlike carbon surfaces with an atomic force microscope. The surface wettability is found to be decisive. Partially hydrophilic surfaces show a logarithmic decrease of friction with increasing velocity, the slope of which varies drastically with humidity, whereas on partially hydrophobic surfaces we confirm the formerly reported logarithmic increase. A model for the thermally activated nucleation of water bridges between tip and sample asperities fully reproduces the experimental data.


Scanning electron microscopy of porcine jejunal loops infused with Escherichia coli endotoxin.
Phenol-extracted Escherichia coli endotoxin was infused into ligated jejunal loops in 3 pigs. Tissue samples were collected from the center of each loop 6 hours later and examined with scanning electron microscopy. Control loops infused with pyrogen-free water were examined, as well as nonligated biopsy and blind-loop samples. Loops infused with endotoxin had predominantly ridge-shaped villi, many actively secreting goblet cells, and numerous large epithelial ulcerations involving the tips and sides of villi. Tight junction disruption between epithelial cells was observed in 1 pig. In contrast, small ulcers were rarely seen in ligated control loops. Villi were predominantly tongue-shaped, and goblet cell activity was moderate. Pathologic changes were greater in endotoxin-infused loops than in water-infused loops at 6 hours after infusion.