Separate the Sheep from the Goats: Use and Limitations of Large Animal Models in Intervertebral Disc Research.
Approximately 5,168 large animals (pigs, sheep, goats, and cattle) were used for intervertebral disc research in identified studies published between 1985 and 2016. Most of the reviewed studies revealed a low scientific impact, a lack of sound justifications for the animal models, and a number of deficiencies in the documentation of the animal experimentation. The scientific community should take suitable measures to investigate the presumption that animal models have translational value in intervertebral disc research. Recommendations for future investigations are provided to improve the quality, validity, and usefulness of animal studies for intervertebral disc research. More in vivo studies are warranted to comprehensively evaluate the suitability of animal models in various applications and help place animal models as an integral, complementary part of intervertebral disc research.


[Correction of immunological status of patients with cholelithiasis for the prevention of postoperative complications].
An investigation of immune status of patients with chronic calculous cholecystitis prior to operation and in the postoperative period has proved the development of a pronounced secondary immune deficiency involving T and B-systems of the immune control as well as factors of nonspecific resistance. These changes may be considered as risk factors of the development of infectious complications in the postoperative period. Prophylactics of the secondary immune deficiency by administration of Contrical is indicated which permitted the amount of postoperative complications and the period of stay at the hospital to be decreased.


Conventional MRI evaluation of gliomas.
MRI using T(1) weighted, T(2) weighted and gadolinium-enhanced sequences plays a central clinical role in diagnosis, characterisation, surveillance and therapeutic monitoring of gliomas. Such conventional MRI protocols provide high resolution multiplanar structural information, and substantially improved tissue characterisation compared with CT. However, the MRI signal lacks biological specificity, e.g. T(2) weighted dependent signal abnormality is dominated by tissue water content, and contrast enhancement reflects a non-specific increase in blood-brain barrier permeability. This limits non-invasive glioma diagnosis, characterisation and therapeutic planning and assessment of active tumour load may be confounded by treatment-related effects. The complex features of glioma morphology and often subtle changes between MRI examinations are also frequently difficult to detect reliably by visual inspection of the images, even by an experienced radiologist. Moreover, the most widely used response criteria in clinical practice and therapeutic trials rely on linear measurements of enhancing tumour and are further challenged by the irregular shape and heterogeneous composition of gliomas. This contributes to the poor correlation of these criteria with hard clinical endpoints. While conventional MRI is widely available and provides essential anatomical information, the lack of pathology-specific biomarkers available from standard MRI sequences and methods of image analysis used limit overall diagnostic and prognostic efficacy of the examination.


A Dual Repeat Cis-Element Determines Expression of GERANYL DIPHOSPHATE SYNTHASE for Monoterpene Production in Phalaenopsis Orchids.
Phalaenopsis bellina is a scented orchid emitting large amount of monoterpenes. GERANYL DIPHOSPHATE SYNTHASE (PbGDPS) is the key enzyme for monoterpene biosynthesis, and shows concomitant expression with the emission of monoterpenes during flower development in P. bellina. Here, we identified a dual repeat cis-element in the GDPS promoter that is critical for monoterpene biosynthesis in Phalaenopsis orchids. A strong correlation between the dual repeat and the monoterpene production was revealed by examination of the GDPS promoter fragments over 12 Phalaenopsis species. Serial-deletion of the 2-kb GDPS promoter fragments demonstrated that the integrity of the dual repeat was crucial for its promoter activities. By screening the Arabidopsis transcription factors (TFs) cDNA library using yeast one-hybrid assay, AtbZIP18, a member of group I of bZIP TFs, was identified to be able to bind the dual repeat. We then identified PbbZIP4 in the transcriptome of P. bellina, showing 83% identity in the DNA binding region with that of AtbZIP18, and the expression level of PbbZIP4 was higher in the scented orchids. In addition, PbbZIP4 transactivated the GDPS promoter fragment containing the dual repeat in dual luciferase assay. Furthermore, transient ectopic expression of PbbZIP4 induced a 10-fold production of monoterpenoids in the scentless orchid. In conclusion, these results indicate that the dual repeat is a real TF-bound cis-element significant for GDPS gene expression, and thus subsequent monoterpene biosynthesis in the scented Phalaenopsis orchids.


Outliers in SAR and QSAR: is unusual binding mode a possible source of outliers?
A lead optimization is usually carried out by structure-activity relationship (SAR) and/or quantitative structure-activity relationship (QSAR) studies. One of the assumptions in SAR and QSAR studies is that similar analogs bind to the same binding site in a similar binding mode. One often observes that there are outliers, especially in QSAR. However, most QSAR studies are carried out focusing their attention to the development of QSAR and leave the outliers without much attention. We searched a number of ligand-bound X-ray crystal structures from the protein structure database to find evidences that could indicate a possible source of outliers in SAR or QSAR. Our results show that unusual binding mode could be a source of outliers.


Prevalence of Bilateral JOCD of the Knee and Associated Risk Factors.
Juvenile osteochondritis dissecans (JOCD) of the knee affects cartilage and subchondral bone surface. Multifocal JOCD is described as multiple lesions within the knee or presence of lesions in other joints. The true prevalence of bilaterality of JOCD is unknown. The purpose of this study is to determine the prevalence of bilateral JOCD and to identify potential risk factors for bilateral disease. We evaluated 108 consecutive patients presenting for JOCD at a single pediatric hospital system. If an OCD lesion of the knee was found, contralateral knee x-rays were performed. Lesion location was documented according to Cahill and Berg, magnetic resonance imaging (MRI) grading documented according to Dipaola, and if surgical treatment was undertaken, intraoperative grading performed according to Guhl. Patients with unilateral JOCD were compared with those with bilateral disease. Statistical analysis of categorical data was performed utilizing likelihood ratio χ test or Fisher exact test and continuous data compared using nonparametric Wilcoxon 2-sample test. There were 85 male (79%) and 23 females (21%) with an average age of 12.3 years (range, 6 to 18 y). Sixty-three percent of lesions were located on the medial femoral condyle and 33% on the lateral femoral condyle. Ninety percent of all lesions were considered weight-bearing lesions. Eighty percent were considered stable on MRI evaluation. Of those lesions that underwent surgical intervention, 61% were either grade I or II lesions. Seventy-three of 108 patients (68%) underwent some form of surgical intervention. Thirty-one patients (29%) were found to have contralateral JOCD lesions. Thirty-nine percent of contralateral lesions found on contralateral radiographs were asymptomatic at presentation and nearly all of those evaluated with MRI (16 of 18) were stable. Sixty-nine percent of contralateral lesions were located on the medial femoral condyle, 27% on the lateral femoral condyle, and 94% were considered weight-bearing lesions. Twelve of 31 contralateral lesions (39%) underwent surgical intervention. Comparing patients with unilateral and bilateral disease, female patients (P<0.05) and younger age at presentation (P<0.009) were risk factors for bilateral JOCD. No statistical difference among other variables was seen with regard to location, MRI or operative stability of lesion, or presence of symptoms. In our consecutive series of 108 patients with JOCD, we found a 29% incidence of bilateral disease. Almost 40% of contralateral lesions were asymptomatic upon presentation. Female sex and younger age at presentation were significant risk factors for bilateral disease. Lesion location, stability, and pain were not statistically significant variables. The authors recommend bilateral radiographic knee evaluation for all patients found to have JOCD. Level IV-retrospective case series.


Studies of the optical properties of metal-pliable polymer composite materials.
Polymer-nano-metallic-particle composites have demonstrated technological potential due to their unique optical and electrical properties. Herein, we report on composites prepared via physical vapor deposition of silver metal onto pliable poly(dimethylsiloxane) (PDMS) polymer. Rapid Ag diffusion and nano-metallic-particle formation in a phase-separated surface layer of the PDMS creates unique sub-surface-based composites whose properties vary based on rate of deposition and average Ag thickness. Additionally, nanometallic-particle spacing can be altered with fair reproducibility and reversibility by physically manipulating the Ag-PDMS composite. The optical properties of the materials are studied by visible wavelength optical extinction spectrometry and surface-enhanced Raman scattering (SERS), including studies performed during physical manipulation. Direct current (DC) conductivity measurements were made during Ag deposition to study percolation conditions for the materials. Depth-profiling was performed by X-ray photoelectron spectrometry. Sample Raman spectral data collected with the composite as a SERS substrate are included. A practical technological characteristic of these composite materials arises from their potential to be molded into functional devices.


Emergency subtotal colectomy in a Jehovah's Witness with massive lower gastrointestinal bleeding: challenges encountered and lessons learned.
A 66-year-old woman who was a Jehovah's Witness had massive lower gastrointestinal bleeding and subsequent hypovolemic shock, necessitating a subtotal colectomy. During the postoperative period, her hemoglobin level decreased to a low of 2.6 g/dL, prolonging her dependence on mechanical ventilation. Prudent perioperative care resulted in a successful outcome. Blood-conserving techniques are indispensable in the management of Jehovah's Witnesses who have massive blood loss. Maximizing oxygen transport, minimizing blood loss, using a cell saver when permissible, providing optimal ventilatory support, performing tracheostomy early if prolonged mechanical ventilation is expected, and augmenting hemoglobin production with administration of iron and erythropoietin are techniques that can facilitate successful outcome in patients who refuse blood transfusion.


The yeast forkhead transcription factors fkh1 and fkh2 regulate lifespan and stress response together with the anaphase-promoting complex.
Forkhead box O (FOXO) transcription factors have a conserved function in regulating metazoan lifespan. A key function in this process involves the regulation of the cell cycle and stress responses including free radical scavenging. We employed yeast chronological and replicative lifespan assays, as well as oxidative stress assays, to explore the potential evolutionary conservation of function between the FOXOs and the yeast forkhead box transcription factors FKH1 and FKH2. We report that the deletion of both FKH genes impedes normal lifespan and stress resistance, particularly in stationary phase cells, which are non-responsive to caloric restriction. Conversely, increased expression of the FKHs leads to extended lifespan and improved stress response. Here we show the Anaphase-Promoting Complex (APC) genetically interacts with the Fkh pathway, likely working in a linear pathway under normal conditions, as fkh1Δ fkh2Δ post-mitotic survival is epistatic to that observed in apc5(CA) mutants. However, under stress conditions, post-mitotic survival is dramatically impaired in apc5(CA) fkh1Δ fkh2Δ, while increased expression of either FKH rescues APC mutant growth defects. This study establishes the FKHs role as evolutionarily conserved regulators of lifespan in yeast and identifies the APC as a novel component of this mechanism under certain conditions, likely through combined regulation of stress response, genomic stability, and cell cycle regulation.


Associations of olfactory dysfunction with anthropometric and cardiometabolic measures: Findings from the 2013-2014 national health and nutrition examination survey (NHANES).
We examined associations of olfactory dysfunction with anthropometric and cardiometabolic measures in a nationally representative sample of US adults. In the 2013-2014 National Health and Nutrition Examination Survey (NHANES), 3,815 participants, 40 years and older, completed a standardized taste and smell protocol, which consisted of an 8-item odor identification test and a chemosensory questionnaire. Measured dysfunction was incorrect identification of ≥3 of 8 odors; self-reported dysfunction was affirmative response to either a recent smell problem, worse ability since age 25, or phantosmia. Survey-weighted linear regression models tested associations of olfactory dysfunction with body mass index (BMI), waist circumference (WC), blood pressure, serum total cholesterol (TC) with fractions, triglycerides, and glucose levels. Models were adjusted for age, race, education, physical activity, self-reported general health condition, smoking history, and income-to-poverty ratio, stratifying by sex and age group (middle-age 40-64 years; older ≥65 years). Relative to normal, measured olfactory dysfunction was associated with lower BMI [β=-1.6 (95% CI: -3.2, -0.01)] in older men. In middle-age women, dysfunction was associated with higher BMI and WC, whether assessed by examination [β's for BMI=3.1 (0.6, 5.5), WC=5.0 (0.3, 9.8)] or self-report [β's for BMI=2.5 (0.6, 4.3), WC=6.1 (2.2, 9.9)]. Measured dysfunction was associated with significantly higher TC [β=12.8 (7.5, 18.1)] and LDL [β=18.1 (9.1, 27.2)] among older men, but significantly lower TC [β=-15.0 (-25.0, -5.7)] and marginally lower LDL [β=-12.0 (-25.0, 1.2)] among older women. Between measured dysfunction and fasting glucose, the association was inverse [β=-7.9 (-13.0, -2.6)] among middle-age men, but positive [β=15.6 (1.5, 29.7)] among older women. No significant associations were observed with blood pressure levels. In conclusion, among US adults ≥40 years, olfactory dysfunction is associated with anthropometric and glucose and lipid levels, with associations varying by sex and age group.


Structures of mithramycin analogues bound to DNA and implications for targeting transcription factor FLI1.
Transcription factors have been considered undruggable, but this paradigm has been recently challenged. DNA binding natural product mithramycin (MTM) is a potent antagonist of oncogenic transcription factor EWS-FLI1. Structural details of MTM recognition of DNA, including the FLI1 binding sequence GGA(A/T), are needed to understand how MTM interferes with EWS-FLI1. We report a crystal structure of an MTM analogue MTM SA-Trp bound to a DNA oligomer containing a site GGCC, and two structures of a novel analogue MTM SA-Phe in complex with DNA. MTM SA-Phe is bound to sites AGGG and GGGT on one DNA, and to AGGG and GGGA(T) (a FLI1 binding site) on the other, revealing how MTM recognizes different DNA sequences. Unexpectedly, at sub-micromolar concentrations MTMs stabilize FLI1-DNA complex on GGAA repeats, which are critical for the oncogenic function of EWS-FLI1. We also directly demonstrate by nuclear magnetic resonance formation of a ternary FLI1-DNA-MTM complex on a single GGAA FLI1/MTM binding site. These biochemical and structural data and a new FLI1-DNA structure suggest that MTM binds the minor groove and perturbs FLI1 bound nearby in the major groove. This ternary complex model may lead to development of novel MTM analogues that selectively target EWS-FLI1 or other oncogenic transcription factors, as anti-cancer therapeutics.


Current status and implications of microRNAs in ovarian cancer diagnosis and therapy.
Ovarian cancer is the fifth most common cancer among women and causes more deaths than any other type of female reproductive cancer. Currently, treatment of ovarian cancer is based on the combination of surgery and chemotherapy. While recurrent ovarian cancer responds to additional chemotherapy treatments, the progression-free interval becomes shorter after each cycle, as chemo-resistance increases until the disease becomes incurable. There is, therefore, a strong need for prognostic and predictive markers to help optimize and personalize treatment in order to improve the outcome of ovarian cancer. An increasing number of studies indicate an essential role for microRNAs in ovarian cancer progression and chemo-resistance. MicroRNAs (miRNAs) are small endogenous non-coding RNAs (~22bp) which are frequently dysregulated in cancer. Typically, miRNAs are involved in crucial biological processes, including development, differentiation, apoptosis and proliferation. Two families of miRNAs, miR-200 and let-7, are frequently dysregulated in ovarian cancer and have been associated with poor prognosis. Both have been implicated in the regulation of epithelial-to-mesenchymal transition, a cellular transition associated with tumor aggressiveness, tumor invasion and chemo-resistance. Moreover, miRNAs also have possible implications for improving cancer diagnosis; for example miR-200 family, let-7 family, miR-21 and miR-214 may be useful in diagnostic tests to help detect ovarian cancer at an early stage. Additionally, the use of multiple target O-modified antagomirs (MTG-AMO) to inhibit oncogenic miRNAs and miRNA replacement therapy for tumor suppressor miRNAs are essential tools for miRNA based cancer therapeutics. In this review we describe the current status of the role miRNAs play in ovarian cancer and focus on the possibilities of microRNA-based therapies and the use of microRNAs as diagnostic tools.


Correlation of genotypes for thiopurine methyltransferase and inosine triphosphate pyrophosphatase with long-term clinical outcomes in Korean patients with inflammatory bowel diseases during treatment with thiopurine drugs.
There is a lack of research describing the associations between thiopurine methyltransferase (TPMT)/inosine triphosphate pyrophosphatase (ITPA) genotypes and long-term clinical outcomes. We investigated whether TPMT/ITPA genotypes predicted long-term clinical response in Korean patients with inflammatory bowel diseases (IBDs) undergoing thiopurine treatment. A total of 204 patients with IBD in whom thiopurine treatment was indicated were enrolled and categorized by TPMT and ITPA genotypes. Long-term follow-up clinical data for these patients were analyzed with specific focus on disease relapse. Of the 204 patients, 162 (79.4%) patients using thiopurines achieved remission and were included in an analysis of long-term clinical outcomes. There were no significant differences in disease relapse-free survival between wild and mutant types of TPMT (P=0.903) or ITPA (P=0.392), according to the results of the log-rank analysis. Our study suggests that TPMT and ITPA genotypes may not affect the rates of disease relapse in IBD patients treated with thiopurines. Further studies are indicated to confirm the utility of TPMT/ITPA genotyping to guide clinicians formulating individualized treatments for IBD patients requiring thiopurine therapy.


Surveillance of sight loss due to delay in ophthalmic treatment or review: frequency, cause and outcome.
PurposeTo determine the frequency of patients suffering harm due to delay in ophthalmic care in the UK over a 12-month period.MethodsPatients with deterioration in vision in at least one eye of 3 lines of Snellen acuity or 15 letters on ETDRS chart or deterioration in visual field deviation of 3 decibels due to health service initiated delay in review or care were ascertained through the BOSU using prospective active surveillance involving all UK consultant ophthalmologists. Demographic details, diagnosis, cause and length of delay, and vision loss were then sought by questionnaire.Results238 cases reported between March 2015 and February 2016. 197/238 questionnaires were returned (83%). Twenty-eight reports were out of the study period or did not meet the case definition. Median age was 76 years (range: 1 to 98 years). Median delay was 22 weeks (range: 2 days to 5½ years). Seventy two per cent experienced permanent reduction in visual acuity, 23% permanent deterioration in visual field. Main diagnoses were Glaucoma 42%, Age-related Macular Degeneration (AMD) 23%, and Diabetic Retinopathy (DR) 16%. Eighteen patients were eligible for Severely Sight Impaired (SSI) or Sight Impaired (SI) registration. Main causes were delayed follow-up (76%), lost referral (7%), and delayed treatment (8%).ConclusionPatients are suffering preventable harm due to health service initiated delay leading to permanently reduced vision. This is occurring in patients of all ages, but most consistently in those with chronic conditions. Delayed follow-up or review is the cause in the majority of cases indicating a lack of capacity within the hospital eye service.


An approach to the patient with unexplained chest pain.
After coronary artery disease has been excluded as a cause for unexplained chest pain, oesophageal disease must be considered. Ambulatory pH monitoring reveals abnormal oesophageal acid exposure in 50% of patients with recurrent non-cardiac chest pain.


Preparation of Selenium-Based Drug-Modified Polymeric Ligand-Functionalised Fe3O4 Nanoparticles as Multimodal Drug Carrier and Magnetic Hyperthermia Inductor.
In recent years, much effort has been invested into developing multifunctional drug delivery systems to overcome the drawbacks of conventional carriers. Magnetic nanoparticles are not generally used as carriers but can be functionalised with several different biomolecules and their size can be tailored to present a hyperthermia response, allowing for the design of multifunctional systems which can be active in therapies. In this work, we have designed a drug carrier nanosystem based on Fe3O4 nanoparticles with large heating power and 4-amino-2-pentylselenoquinazoline as an attached drug that exhibits oxidative properties and high selectivity against a variety of cancer malignant cells. For this propose, two samples composed of homogeneous Fe3O4 nanoparticles (NPs) with different sizes, shapes, and magnetic properties have been synthesised and characterised. The surface modification of the prepared Fe3O4 nanoparticles has been developed using copolymers composed of poly(ethylene-alt-maleic anhydride), dodecylamine, polyethylene glycol and the drug 4-amino-2-pentylselenoquinazoline. The obtained nanosystems were properly characterised. Their in vitro efficacy in colon cancer cells and as magnetic hyperthermia inductors was analysed, thereby leaving the door open for their potential application as multimodal agents.


Sample size estimates for atrial fibrillation endpoints.
This article reviews the current sample size requirements for studies evaluating the efficacy of atrial therapies. Sample sizes for several study designs and endpoints are presented. However, emphasis is given to studies conducted in patients with implantable devices in the light of new available data from recent trials involving such patients. Studies utilizing mortality require >5,000 patients followed for at least 2 years. Symptomatic episode recurrence represents a possible alternate endpoint but still requires >400 patients followed for 1 year. Parallel design studies for device atrial fibrillation (AF) burden and frequency endpoints were found to be of little value because they would require thousands of patients. In contrast, cross-over studies utilizing device AF burden or AF frequency endpoints provide the best option in terms of sample size and study duration. A 30% therapy efficacy (reduction in AF burden or frequency) can be detected with just a little over 100 patients followed for 6 months. Therefore, cross-over studies with device-based endpoints are a viable option to evaluate and screen possible new rhythm control treatments before evaluating their effect on clinical outcome variables that may require larger trials with longer follow-up times.


Grids in topographic maps reduce distortions in the recall of learned object locations.
To date, it has been shown that cognitive map representations based on cartographic visualisations are systematically distorted. The grid is a traditional element of map graphics that has rarely been considered in research on perception-based spatial distortions. Grids do not only support the map reader in finding coordinates or locations of objects, they also provide a systematic structure for clustering visual map information ("spatial chunks"). The aim of this study was to examine whether different cartographic kinds of grids reduce spatial distortions and improve recall memory for object locations. Recall performance was measured as both the percentage of correctly recalled objects (hit rate) and the mean distance errors of correctly recalled objects (spatial accuracy). Different kinds of grids (continuous lines, dashed lines, crosses) were applied to topographic maps. These maps were also varied in their type of characteristic areas (LANDSCAPE) and different information layer compositions (DENSITY) to examine the effects of map complexity. The study involving 144 participants shows that all experimental cartographic factors (GRID, LANDSCAPE, DENSITY) improve recall performance and spatial accuracy of learned object locations. Overlaying a topographic map with a grid significantly reduces the mean distance errors of correctly recalled map objects. The paper includes a discussion of a square grid's usefulness concerning object location memory, independent of whether the grid is clearly visible (continuous or dashed lines) or only indicated by crosses.


Testing virtual reality-based cue-exposure software: Which cue-elicited responses best discriminate between patients with eating disorders and healthy controls?
Virtual reality (VR) technologies have been proposed as a new tool able to improve on in vivo exposure in patients with eating disorders. This study assessed the validity of a VR-based software for cue exposure therapy (CET) in people with bulimia nervosa (BN) and binge eating disorder (BED). Fifty eight outpatients (33 BN and 25 BED) and 135 healthy participants were exposed to 10 craved virtual foods and a neutral cue in four experimental virtual environments (kitchen, dining room, bedroom, and cafeteria). After exposure to each VR scenario, food craving and anxiety were assessed. The frequency/severity of episodes of uncontrollable overeating was also assessed and body mass index was measured prior to the exposure. In both groups, craving and anxiety responses when exposed to the food-related virtual environments were significantly higher than in the neutral-cue virtual environment. However, craving and anxiety levels were higher in the clinical group. Furthermore, cue-elicited anxiety was better at discriminating between clinical and healthy groups than cue-elicited craving. This study provides evidence of the ability of food-related VR environments to provoke food craving and anxiety responses in BN and BED patients and highlights the need to consider both responses during treatment. The results support the use of VR-CET in the treatment of eating disorder patients characterized by binge-eating and people with high bulimic symptoms.


Testing the effects of brief intervention in primary care for problem drug use in a randomized controlled trial: rationale, design, and methods.
A substantial body of research has established the effectiveness of brief interventions for problem alcohol use. Following these studies, national dissemination projects of screening, brief intervention (BI), and referral to treatment (SBIRT) for alcohol and drugs have been implemented on a widespread scale in multiple states despite little existing evidence for the impact of BI on drug use for non-treatment seekers. This article describes the design of a study testing the impact of SBIRT on individuals with drug problems, its contributions to the existing literature, and its potential to inform drug policy. The study is a randomized controlled trial of an SBIRT intervention carried out in a primary care setting within a safety net system of care. Approximately 1,000 individuals presenting for scheduled medical care at one of seven designated primary care clinics who endorse problematic drug use when screened are randomized in a 1:1 ratio to BI versus enhanced care as usual (ECAU). Individuals in both groups are reassessed at 3, 6, 9, and 12 months after baseline. Self-reported drug use and other psychosocial measures collected at each data point are supplemented by urine analysis and public health-related data from administrative databases. This study will contribute to the existing literature by providing evidence for the impact of BI on problem drug use based on a broad range of measures including self-reported drug use, urine analysis, admission to drug abuse treatment, and changes in utilization and costs of health care services, arrests, and death with the intent of informing policy and program planning for problem drug use at the local, state, and national levels. ClinicalTrials.gov NCT00877331.


The cleaved-Caspase-3 antibody is a marker of Caspase-9-like DRONC activity in Drosophila.
The cleaved-Caspase-3 antibody is a popular tool in apoptosis research in Drosophila. As the antibody was raised against cleaved human Caspase-3, it was assumed that it detects cleaved DRICE and DCP-1, Caspase-3-like effector caspases in Drosophila. However, as shown here, strong immunoreactivity persists in apoptotic models doubly mutant for drICE and dcp-1. In contrast, mutants of the apoptosome components DRONC (Caspase-9-like) and ARK (Apaf-1 related) do not label with the cleaved-Caspase-3 antibody. By peptide blocking experiments and further genetic studies, we provide evidence that the cleaved-Caspase-3 antibody recognizes multiple proteins including DCP-1 and likely DRICE, but also at least one additional unknown protein, all of which require DRONC for epitope exposure. The unknown substrate may be involved in non-apoptotic functions of DRONC. Because the cleaved-Caspase-3 antibody not only detects cleaved Caspase-3-like proteins in Drosophila, but also other proteins in a DRONC-dependent manner, it is more accurate to consider the cleaved-Caspase-3 antibody as a marker for DRONC activity, rather than effector caspase activity.


[Propoleum and Peyronie's disease].
Peyronie's disease is a plastic penile induration which represents a well-known problem affecting men in their middle age, who suffer penile incurvation during erections, pain and poor consistency at the site of incurvation. There is no satisfactory treatment for this disease. A patient with Peyronie's disease who received propoleum treatment for a giardiasis infection referred spontaneous improvement of this penile incurvation as treatment cycles were repeated. So, we decided to treat a group of patients with this pathologic entity with propoleum to evaluate its efficacy. Initially, 25 patients were studied, only 13 completed the study 6 months later. age, penile incurvation, presence or not of pain with erections, detection and measurement (cm) of the fibrous plaque on physical examination and ultrasound, and description of its radiologic characteristics. Propoleum powder (300 mg) was administered for 6 months. Patient age was predominantly between 51 and 60 years. Upward deviation was the most frequent presentation; it disappeared in 2 patients in the "Improved" group. Most patients had no pain. 3 patients out of 13 had no plaque on examination at the end of treatment. Regarding incurvation 10 patients improved, 2 improved slightly, and only one improved somewhat during the first three months but then after remained the same. Penile x-ray was not useful as a diagnostic test; penile ultrasound was greatly useful. Penile incurvation improved in 77% of the patients. Fibrous plaques were reduced more than 0.5 cm on ultrasound (average 0.64 cm).


Anxiety and depression in working-age cancer survivors: a register-based study.
Anxiety and depression can be a long-term strain in cancer survivors. Little is known about the emotional situation of cancer survivors who have to deal with work- and family-related issues. The purpose of this study was to investigate anxiety and depression in working-age cancer survivors and associated factors. A register-based sample of 3370 cancer survivors (25 to 55 years at time of diagnosis) diagnosed up to six years prior to the survey was recruited from two German cancer registries. Demographic and medical characteristics as well as self-reported measures were used. Overall, approximately 40% of the survivors reported moderate to high anxiety scores and approximately 20% reported moderate to high depression scores. Compared to the general population, working-age cancer survivors were more anxious but less depressed (p < .001). Subgroups with regard to time since diagnosis did not differ in anxiety or depression. Anxiety and depression in cancer survivors were associated with various variables. Better social support, family functioning and physical health were associated with lower anxiety and depression. Overall, we found higher anxiety levels in cancer survivors of working-age than in the general population. A considerable portion of cancer survivors reported moderate to high levels of anxiety and depression. The results indicate the need for psychosocial screening and psycho-oncological support e.g. in survivorship programs for working-age cancer survivors. Assessing the physical health, social support and family background might help to identify survivors at risk for higher emotional distress.


The Contextual Association Network Activates More for Remembered than for Imagined Events.
The human capacities to remember events from the past and imagine events in the future rely on highly overlapping neural substrates. Neuroimaging studies have revealed brain regions that are more active for imagined events than remembered events, but the reverse pattern has not been shown consistently. Given that remembered events tend to be associated with more contextual information ( Johnson et al. 1988), one might expect a set of regions to demonstrate greater activity for remembered events. Specifically, regions sensitive to the strength of contextual associations might be hypothesized to show greater activity for remembered events. The present experiment tests this hypothesis. fMRI was used to identify brain regions within the contextual association network ( Bar and Aminoff 2003); regions within this network were then examined to see whether they showed differential activity during remembering and imagining. Bilateral regions within the parahippocampal cortex and retrosplenial complex responded more strongly to remembered past events, supporting work that suggests these events have more contextual information associated with them. Follow-up voxel-wise analysis demonstrated the specificity of these results, as did re-analysis of previous experimental datasets. These results suggest that a key differentiating feature of remembering and imagining is the strength of contextual associations.


Ongoing Transposon-Mediated Genome Reduction in the Luminous Bacterial Symbionts of Deep-Sea Ceratioid Anglerfishes.
Diverse marine fish and squid form symbiotic associations with extracellular bioluminescent bacteria. These symbionts are typically free-living bacteria with large genomes, but one known lineage of symbionts has undergone genomic reduction and evolution of host dependence. It is not known why distinct evolutionary trajectories have occurred among different luminous symbionts, and not all known lineages previously had genome sequences available. In order to better understand patterns of evolution across diverse bioluminescent symbionts, we de novo sequenced the genomes of bacteria from a poorly studied interaction, the extracellular symbionts from the "lures" of deep-sea ceratioid anglerfishes. Deep-sea anglerfish symbiont genomes are reduced in size by about 50% compared to free-living relatives. They show a striking convergence of genome reduction and loss of metabolic capabilities with a distinct lineage of obligately host-dependent luminous symbionts. These losses include reductions in amino acid synthesis pathways and abilities to utilize diverse sugars. However, the symbiont genomes have retained a number of categories of genes predicted to be useful only outside the host, such as those involved in chemotaxis and motility, suggesting that they may persist in the environment. These genomes contain very high numbers of pseudogenes and show massive expansions of transposable elements, with transposases accounting for 28 and 31% of coding sequences in the symbiont genomes. Transposon expansions appear to have occurred at different times in each symbiont lineage, indicating either independent evolutions of reduction or symbiont replacement. These results suggest ongoing genomic reduction in extracellular luminous symbionts that is facilitated by transposon proliferations.IMPORTANCE Many female deep-sea anglerfishes possess a "lure" containing luminous bacterial symbionts. Here we show that unlike most luminous symbionts, these bacteria are undergoing an evolutionary transition toward small genomes with limited metabolic capabilities. Comparative analyses of the symbiont genomes indicate that this transition is ongoing and facilitated by transposon expansions. This transition may have occurred independently in different symbiont lineages, although it is unclear why. Genomic reduction is common in bacteria that only live within host cells but less common in bacteria that, like anglerfish symbionts, live outside host cells. Since multiple evolutions of genomic reduction have occurred convergently in luminous bacteria, they make a useful system with which to understand patterns of genome evolution in extracellular symbionts. This work demonstrates that ecological factors other than an intracellular lifestyle can lead to dramatic gene loss and evolutionary changes and that transposon expansions may play important roles in this process.


Expression and identification of recombinant soluble single-chain variable fragment of monoclonal antibody MC3.
To generate soluble single chain variable fragments (ScFv) of monoclonal antibody MC3 recognizing colorectal and gastric carcinomas. mRNA was isolated from the hybridoma cell line producing MC3 and the DNAs encoding variable domains of heavy and light chains (VH and VL) of the antibody were amplified separately by RT-PCR and assembled into ScFv DNA with a linker DNA. The ScFv DNA was ligated into the phagemid vector pCANTAB5E and the ligated sample was transformed into E.coli TG1. The transformed cells were infected with M13KO7 helper phage to yield recombinant phages. After two rounds of panning with gastric carcinoma cell line AGS highly expressing MC3-binding antigen, the phage clones displaying ScFv fragments of the antibody were selected by ELISA. 4 phage clones showing strong signal in ELISA were used to infect E.coli HB2151 to express soluble ScFvs. The soluble ScFvs were identified by Dot blot and Western blot, and their antigen-binding activity was assayed by ELISA. The VH and VL DNAs of the ScFv DNA derived from phage clone 19 were sequenced. The VH,VL and ScFv DNAs were about 340 bp, 320 bp and 750 bp respectively. After two rounds of panning to the recombinant phages, 18 antigen-positive phage clones were selected from 30 preselected phage clones by ELISA. All the soluble ScFvs derived from the 4 out of the 18 antigen-positive phage clones were about M(r)32000 and concentrated in periplasmatic space under the given culture condition. The soluble ScFvs could bind the antigen, and they shared the same binding site with MC3. The sequences of the VH and VL DNAs of the MC3 ScFv showed that the variable antibody genes belonged to the IgG1 subgroup,kappa-type. The soluble ScFv of MC3 is successfully produced, which not only provides a possible novel targeting vehicle for in vivo and in vitro study on associated cancers, but also offers the antibody a stable genetic source.


Retinal oximetry and fractal analysis of capillary maps in sickle cell disease patients and matched healthy volunteers.
Fractal analysis can be used to quantitatively analyze the retinal microvasculature and might be a suitable method to quantify retinal capillary changes in sickle cell disease (SCD) patients. Retinal oximetry measurements might function as a proxy for the pathophysiology of cerebrovascular diseases. Moreover, hypoxia has an important role in the pathophysiology of diabetic and other retinopathies. However, little is known about the oximetry around the macula in SCD patients. With this study, we explored the feasibility to perform these quantified measurements in SCD patients. Retinal microvascular and oximetry measurements were performed in eight SCD patients and eight healthy matched controls. Oximetry pictures and non-invasive capillary perfusion maps (nCPM) were obtained by the retinal function imager. Measurements were conducted twice on two different study days. Measured variables included monofractal dimension (Dbox), relative saturation, deoxygenated hemoglobin (deoxyHb), and oxygenated hemoglobin (oxyHb) concentration. No statistically significant differences in vessel density were found in the different annular zones (large vessels, p = 0.66; small vessels, p = 0.66) and anatomical quadrants (large vessels, p = 0.74; small vessels, p = 0.72). Furthermore, no significant between-group differences were found in the other different anatomical quadrants and annular zones around the fovea for relative saturation levels and deoxygenated Hb. However, the oxyHb levels were significantly lower in SCD patients, compared with those in matched controls in the temporal quadrants (p = 0.04; p = 0.02) and the superior nasal quadrant (p = 0.05). Our study demonstrated the feasibility of multispectral imaging to measure retinal changes in oxygenation in both SCD patients and matched volunteers. The results suggest that in SCD patients before any structural microvascular changes in the central retina are present, functional abnormalities can be observed with abnormal oximetry measurements.


Human Tissue Processing and Transplantation in MESOT States: How to Promote?
Middle East, with more than 650 million inhabitants, has one the lowest mean donation rate in the world in the context of organ procurement from deceased donors with <2 per million population as actual deceased organ donation rate in 2014. Tissue processing and transplantation in this region are also restricted to a few countries among those representing Middle East Society for Organ Transplantation. Aiming to promote human tissue processing and transplantation in this region, as a life-enhancing therapeutic modality, we have to share our know-how and facilities in order so that the patients in different countries gain almost equal benefit of this vital procedure. To take different infrastructure of Middle Eastern countries into consideration and their plans to extend their activities, this intention will be come to the reality and materialized by primarily sharing the processed tissues among member states in a centralized manner and to offer processing services and banking to locally retrieved tissues.


Long-term care: a substantive factor in financial planning.
More than 50 percent of women will enter a nursing home at some point in their lives. About one-third of men living to age 65 will also need nursing home care. Planning for long-term care is even more important since Medicare covers very little of the cost of such care. The Indiana Partnership Plan is one program designed to help fund the long-term care costs while allowing individuals protect other financial assets.


A data-driven approach to quality risk management.
An effective clinical trial strategy to ensure patient safety as well as trial quality and efficiency involves an integrated approach, including prospective identification of risk factors, mitigation of the risks through proper study design and execution, and assessment of quality metrics in real-time. Such an integrated quality management plan may also be enhanced by using data-driven techniques to identify risk factors that are most relevant in predicting quality issues associated with a trial. In this paper, we illustrate such an approach using data collected from actual clinical trials. Several statistical methods were employed, including the Wilcoxon rank-sum test and logistic regression, to identify the presence of association between risk factors and the occurrence of quality issues, applied to data on quality of clinical trials sponsored by Pfizer. ONLY A SUBSET OF THE RISK FACTORS HAD A SIGNIFICANT ASSOCIATION WITH QUALITY ISSUES, AND INCLUDED: Whether study used Placebo, whether an agent was a biologic, unusual packaging label, complex dosing, and over 25 planned procedures. Proper implementation of the strategy can help to optimize resource utilization without compromising trial integrity and patient safety.


[The characteristics of the development in houseflies of resistance to synergized and nonsynergized permethrin].
Peculiarities of the development of room fly resistance during selection with permetrine have been studied in the laboratory conditions. A strain possessing a 50-fold level of resistance to permetrine has been obtained. A range of cross-resistance of this strain to insecticides of various chemical classes has been determined. It has been established that the basic mechanism regulating resistance to permetrine is an increase in cytochrome P-450 monooxygenases. Resistance can be overcome using a mixture consisting of permetrine and monooxygenase inhibitor piperonylbutoxide. The mixture retains its efficacy in almost 20 generations; later on the development of resistance to the mixture is accompanied by an increase in the level of resistance to nonsynergized piretroides and other preparations.


[pH-dependence of inhibitory action of eosin Y on ATPase activity of smooth muscle actomyosin complex of the uterus].
Research of pH-dependence of inhibitory action of eosin Y (2',4',5',7'-tetrabromofluorescin) on ATPase of contractile proteins of smooth muscles of the uterus has shown that the increase of concentration of this inhibitor (from 0.1 to 10 microM) influenced the profile of pH-dependence of ATPase activity of actomyosin: in the presence of 0.1 microM eosin Y the change of optimal value of pH has been observed in more sour side in relation to the control; at the increase of concentration of eosin Y (from 0.5 to 10 microM) the strongly pronounced optimum of pH is absents in general. The ability of eosin Y to inhibit the ATPase activity of contractile complex is dependent on pH of incubation environment. The change of pH from 6.0 to 7.2 results in a 9-fold decrease of magnitude of apparent constant of inhibition Ki (from 6.5 +/- 0.8 microM to 0.74 +/- 0.07 microM). The obtained results indicate that the diminishing of concentration of H+ in an incubation environment favors the increase of affinity ATPase of actomyosin for eosin Y and prove the important role of ionization processes in the system "enzyme-substrate-inhibitor" for realization of inhibitory action of eosin Y.


A differential diagnosis between uterine leiomyoma and leiomyosarcoma using transcriptome analysis.
The objective of this study was to estimate the accuracy of transcriptome-based classifier in differential diagnosis of uterine leiomyoma and leiomyosarcoma. We manually selected 114 normal uterine tissue and 31 leiomyosarcoma samples from publicly available transcriptome data in UCSC Xena as training/validation sets. We developed pre-processing procedure and gene selection method to sensitively find genes of larger variance in leiomyosarcoma than normal uterine tissues. Through our method, 17 genes were selected to build transcriptome-based classifier. The prediction accuracies of deep feedforward neural network (DNN), support vector machine (SVM), random forest (RF), and gradient boosting (GB) models were examined. We interpret the biological functionality of selected genes via network-based analysis using GeneMANIA. To validate the performance of trained model, we additionally collected 35 clinical samples of leiomyosarcoma and leiomyoma as a test set (18 + 17 as 1st and 2nd test sets). We discovered genes expressed in a highly variable way in leiomyosarcoma while these genes are expressed in a conserved way in normal uterine samples. These genes were mainly associated with DNA replication. As gene selection and model training were made in leiomyosarcoma and uterine normal tissue, proving discriminant of ability between leiomyosarcoma and leiomyoma is necessary. Thus, further validation of trained model was conducted in newly collected clinical samples of leiomyosarcoma and leiomyoma. The DNN classifier performed sensitivity 0.88, 0.77 (8/9, 7/9) while the specificity 1.0 (8/8, 8/8) in two test data set supporting that the selected genes in conjunction with DNN classifier are well discriminating the difference between leiomyosarcoma and leiomyoma in clinical sample. The transcriptome-based classifier accurately distinguished uterine leiomyosarcoma from leiomyoma. Our method can be helpful in clinical practice through the biopsy of sample in advance of surgery. Identification of leiomyosarcoma let the doctor avoid of laparoscopic surgery, thus it minimizes un-wanted tumor spread.


Review of complications associated with endoscopic pancreatic cyst-gastrostomy: a single-institution experience.
Pancreatic fluid collections can form after episodes of pancreatitis, either acute or chronic. The majority will resolve spontaneously but when decompression is mandated, endoscopic drainage is the method of choice. However, it is not void of complications. We retrospectively reviewed the charts of 65 patients who underwent endoscopic drainage of pancreatic fluid collections in our institution. The primary outcomes examined included the incidence and type of complications associated with the endoscopic approach. Endoscopic ultrasound was utilized in 86.2% and transgastric approach was used in 81.5% of the cases. The complication rate was 17%. Specifically, complications recorded were infection (6%), perforation and acute abdomen necessitating surgical intervention (4.6%), pneumoperitoneum that was managed nonoperatively (3%), upper gastrointestinal bleed in the knife puncture site that resolved spontaneously (1.5%), and stent migration (1.5%). One patient died remotely to the endoscopic drainage after paracentesis of ascites that resulted in hemorrhagic shock. This study is one of the largest studies reporting the associated morbidity and mortality after endoscopic cyst-gastrostomy. Major and minor complications occurred at a rate of 17% in our study. Endoscopic approach is a safe draining method and should remain the approach of choice for pancreatic fluid collection decompression.


Teratogenic effects of alcohol: a decade of brain imaging.
Heavy alcohol exposure can have serious and long-lasting effects on the developing fetal brain. In the last decade, researchers have utilized quantitative structural magnetic resonance imaging (MRI) to examine the brains of living children and adults with histories of heavy prenatal alcohol exposure. In addition to microcephaly, these studies indicated structural abnormalities in various regions of the brain, including the cerebellum, corpus callosum, and the basal ganglia. Most recently, we have utilized novel imaging and analytic techniques to study the brain as a whole in an effort to elucidate more subtle differences than was possible with earlier techniques. Results indicated displacements in the corpus callosum, increased gray matter densities in both hemispheres in the perisylvian regions, and altered gray matter asymmetry in portions of the temporal lobes in the brains of alcohol-exposed subjects. In addition, prominent shape abnormalities were observed in the brains of these subjects, with narrowing in the temporal region and reduced brain growth in portions of the frontal lobe. These results imply that brain growth continues to be adversely affected long after the prenatal insult and that the brain regions most affected may be consistent with the neurocognitive deficits characteristic of individuals prenatally exposed to alcohol.


Adolescent Mental Health Literacy: Young People's Knowledge of Depression and Social Anxiety Disorder.
Understanding why nearly 80% of youth ages 6-18 years with a mental disorder fail to receive treatment represents an important public health priority. International data suggest that underrecognition of mental illness and the need for treatment are barriers to service utilization. This study extends work to a U.S. sample of 1,104 adolescents. High School students were invited to participate in a self-report study assessing knowledge and beliefs regarding mental illness. Participants completed the survey in groups at school and read vignettes portraying peers experiencing major depression, social anxiety disorder, and a situation where the individual has to cope with a common life stressor followed by a series of questions in reference to each vignette. Adolescents had better recognition of depression than social anxiety disorder and were more likely to recommend seeking help for it. However, <50% of youth recognized depression. Family, friends, and counselors were recommended as sources of help. Differences according to the sex of the respondent and person in the vignette were observed. These data are among the first to provide information regarding the mental health literacy of American adolescents and suggest potential points for intervention. Pending replication of the findings herein, efforts to help adolescents recognize mental health problems and to increase the likelihood of recommending professional help will be important.


Neurophysiological correlates of executive function: a comparison of European-canadian and chinese-canadian 5-year-old children.
This study explored the neurophysiological correlates of executive function (EF) in young children from two different cultural backgrounds. Twenty European-Canadian and 17 Chinese-Canadian 5-year-olds participated in a go/no-go task, during which high-density electroencephalographic (EEG) data were recorded. No cultural group differences were observed in children's behavioral performance on the task, but marked differences were revealed by ERP analyses, which focused on the amplitude and latency of the N2 waveform. Chinese-Canadian children showed larger (i.e., more negative) N2 amplitudes than European-Canadian children on the right side of the scalp on no-go trials, as well as on the left side of the scalp on go trials, and for all children, larger N2 amplitudes were associated with faster median reaction times. Source analyses of the N2 were consistent with the hypothesis that compared to European-Canadian children, Chinese-Canadian children showed more activation in dorsomedial, ventromedial, and (bilateral) ventrolateral prefrontal cortex. These findings reveal that EEG can provide a measure of cultural differences in neurocognitive function that is more sensitive than behavioral data alone; that Chinese-Canadian children show a pattern of hemispheric differentiation in the context of this task than that is more pronounced than that of age-matched European-Canadian children; that the asymmetrically lateralized N2 may be a reliable marker of both effortful inhibition (on the right) and effortful approach (on the left); and that the neural correlates of EF may vary across samples of healthy participants, even in children.


Single-Cell Hemoprotein Diet Changes Adipose Tissue Distributions and Re-Shapes Gut Microbiota in High-Fat Diet-Induced Obese Mice.
We have previously observed that feeding with single-cell hemoprotein (heme-SCP) in dogs (1 g/day for 6 days) and broiler chickens (1 ppm for 32 days) increased the proportion of lactic acid bacteria in the gut while reducing their body weights by approximately 1~2%. To define the roles of heme-SCP in modulating body weight and gut microbiota, obese C57BL/6N mice were administered varied heme-SCP concentrations (0, 0.05, and 0.5% heme-SCP in high fat diet) for 28 days. The heme-SCP diet seemed to restrain weight gain till day 14, but the mice gained weight again later, showing no significant differences in weight. However, the heme-SCP-fed mice had stiffer and oilier bodies compared with those of the control mice, which had flabby bodies and dull coats. When mice were dissected at day 10, the obese mice fed with heme-SCP exhibited a reduction in subcutaneous fat with an increase in muscle mass. The effect of heme-SCP on the obesity-associated dyslipidemia tended to be corroborated by the blood parameters (triglyceride, total cholesterol, and C-reactive protein) at day 10, though the cor-relation was not clear at day 28. Notably, the heme-SCP diet altered gut microbiota, leading to the proliferation of known anti-obesity biomarkers such as Akkermansia, Alistipes, Oscillibacter, Ruminococcus, Roseburia, and Faecalibacterium. This study suggests the potential of heme-SCP as an anti-obesity supplement, which modulates serum biochemistry and gut mi-crobiota in high-fat diet-induced obese mice.


Surviving the NCLEX Dip.
Schools of nursing are charged with preparing graduates to meet minimal practice standards upon workforce entry. To that end, students must pass the National Council of State Boards of Nursing Licensure Examination (NCLEX-RN®) upon graduation. Increased rigor and higher recent passing standards for the exam can contribute to declining scores, which negatively impact graduates and the nursing program. The purpose of this article is to describe a successful 10-step action plan implemented to remediate a brief dip in NCLEX-RN pass rates at one university. The plan contributed to a 10 percent increase in NCLEX scores within one year.


Result of endoscopic biliary drainage in hilar cholangiocarcinoma.
Patients with hilar obstruction usually require bilateral biliary drainage. The prognosis of patients who fail bilateral biliary drainage after contrast injection into both intrahepatic ducts is poor due to a high infection rate in the undrained segments. The incidence of post-endoscopic retrograde cholangiopancreatography cholangitis in those with successful bilateral biliary drainage was less, but still significant. Incomplete subsegmental intrahepatic duct drainage is suggested to be responsible for post-biliary drainage cholangitis in cases of advanced hilar tumors. This study was undertaken to determine the incidence of post-endoscopic retrograde cholangiopancreatography cholangitis, jaundice resolution, and stent clogging in different types of malignant biliary obstruction after biliary drainage. From our endoscopic retrograde cholangiopancreatography database, there were 63 patients who underwent endoscopic biliary drainage between September 2000 and November 2001, for malignant biliary obstruction. Sixty-one endoscopic retrograde cholangiopancreatographies had biliary drainage performed (2 patients who failed biliary drainage were excluded). We divided our patients into 3 groups: Group 1 = Bismuth I, Group 2 = Bismuth II, and Group 3 = Bismuth III and IV. All but 2 Group 1 patients had successful biliary endoprosthesis (plastic [n = 13], metallic [n = 12], failed [n = 2]) placement into an extrahepatic duct. All patients from Group 2 (n = 10) and 20 patients from Group 3 (n = 26) had successful bilateral biliary drainage. Unilateral biliary drainage was performed in 6 patients from Group 3, each with a plastic endoprosthesis. The incidence of post-biliary drainage cholangitis (new onset of fever >38.5 degrees C with leukocytosis), jaundice resolution (normal bilirubin level), and the duration of endoprosthesis patency were compared among the 3 groups. The incidences of post-endoscopic retrograde cholangiopancreatography cholangitis, jaundice resolution, and the duration of endoprosthesis patency were: Group 1 (4%, 96%, and 87.2 days, respectively), Group 2 (10%, 100%, and 69.1 days, respectively) and Group 3 (57.7%, 73.1%, and 41.3 days, respectively). Of those patients who did not undergo surgery, patients from Group 3 required endoprosthesis exchange sooner than others. The outcome of biliary drainage in patients with advanced hilar tumors (Bismuth III or IV) was poorer than hilar tumor at earlier stages (Bismuth I or II).


[Mutagenesis, induced by phosphotriester analogs of oligonucleotides and directed to the cleavage site of double-spiral DNA].
The mutagenic properties of phosphotriester analogues revealed in course of interaction with linearized plasmid DNA were studied. The plasmid-based model system permitting one to test reliably the induced mutations is proposed. The efficiency of mutagenesis was shown to depend on the length of the oligonucleotide-mutagen and the genotype of the transformed Escherichia coli strain. The possible mechanisms involved in mutagenesis are discussed.


Controlled incremental filtration: a simplified approach to design and fabrication of high-throughput microfluidic devices for selective enrichment of particles.
The number of microfluidic strategies aimed at separating particles or cells of a specific size within a continuous flow system continues to grow. The wide array of biomedical and other applications that would benefit from successful development of such technology has motivated the extensive research in this area over the past 15 years. However, despite promising advancements in microfabrication capabilities, a versatile approach that is suitable for a large range of particle sizes and high levels of enrichment, with a volumetric throughput sufficient for large-scale applications, has yet to emerge. Here we describe a straightforward method that enables the rapid design of microfluidic devices that are capable of enriching/removing particles within a complex aqueous mixture, with an unprecedented range of potential cutoff diameter (below 1 μm to above 100 μm) and an easily scalable degree of enrichment/filtration (up to 10-fold and well beyond). A simplified model of a new approach to crossflow filtration - controlled incremental filtration - was developed and validated for its ability to generate microfluidic devices that efficiently separate particles on the order of 1-10 μm, with throughputs of tens of μL min(-1), without the use of a pump. Precise control of the amount of fluid incrementally diverted at each filtration "gap" of the device allows for the gap size (~20 μm) to be much larger than the particles of interest, while the simplicity of the model allows for many thousands of these filtration points to be readily incorporated into a desired device design. This new approach should enable truly high-throughput microfluidic particle-separation devices to be generated, even by users only minimally experienced in fluid mechanics and microfabrication techniques.


Gene structure and expression study of the SEDL gene for spondyloepiphyseal dysplasia tarda.
Spondyloepiphyseal dysplasia tarda (SEDL) is an X-linked recessive disorder of endochondral bone formation caused by mutations in the SEDL gene. Here we present the structural analysis and subcellular localization of human SEDL. The SEDL gene is composed of six exons and spans a genomic region of approximately 20 kb in Xp22. It contains four Alu sequences in its 3' UTR and an alternatively spliced MER20 sequence in its 5' UTR (exon 2). Complex alternative splicing was detected for exon 4. Altogether seven SEDL pseudogenes were detected in the human genome: SEDLP1, a transcribed retropseudogene (or retro-xaptonuon) on chromosome 19q13.4 with potential to encode a protein identical to that of the SEDL gene; SEDLP2, another retropseudogene (not transcribed) on chromosome 8; and five truncated pseudogenes, SEDLP3-SEDLP7, on chromosome Yq11.23. Based on the knowledge of the yeast SEDL ortholog we speculated that the SEDL protein may participate along the ER-to-Golgi transport compartments. To test this hypothesis we performed transient transfection studies with tagged recombinant mammalian SEDL proteins in Cos-7 cells. The tagged SEDL proteins localized to perinuclear structures that partly overlapped with the intermediate ER-Golgi compartment (ERGIC; or vesicular tubular complex, VTC). Two human SEDL mutations (157-158delAT and C271T(STOP)) introduced into SEDL FLAG and GFP constructs led to the misplacement of the SEDL protein primarily to the cell nucleus and partially to the cytoplasm. Based on these experiments we suggest that the COOH end of the SEDL protein might be responsible for proper targeting of SEDL along the ER-Golgi membrane compartments (including Golgi and ERGIC/VTC).


Axl receptor tyrosine kinase is a novel target of apigenin for the inhibition of cell proliferation.
The Axl receptor tyrosine kinase (RTK), along with Tyro 3 and Mer, belongs to the TAM subfamily that promotes survival, stimulates proliferation and/or inhibits apoptosis. In various types of human cancer, including breast, lung and prostate cancer, Axl expression is increased and correlates with an advanced clinical stage. In this study, we examined whether apigenin has an effect on Axl expression, which in turn can affect cell proliferation. The treatment of the non‑small cell lung cancer (NSCLC) cells, A549 and H460, with apigenin decreased Axl mRNA and protein expression in a dose‑dependent manner. Axl promoter activity was also inhibited by apigenin, indicating that apigenin suppressed Axl expression at the transcriptional level. Upon treatment with apigenin, the viability of both the A549 and H460 cells was gradually decreased and the anti-proliferative effects were further confirmed by the dose‑dependent decrease in the clonogenic ability of the apigenin‑treated cells. Subsequently, we found that the viability and clonogenic ability of the cells treated with apigenin was less or more affected by transfection of the cells with a Axl-expressing plasmid or Axl targeting siRNA, compared to transfection with the empty vector or control siRNA, respectively. In addition, apigenin increased the expression of p21, a cyclin-dependent kinase inhibitor, but reduced the expression of X-linked inhibitor of apoptosis protein (XIAP). These cell cycle arrest and pro-apoptotic effects of apigenin were also attenuated or augmented by the up- or downregulation of Axl expression, respectively, which suggests that Axl is a novel target of apigenin through which it exerts its inhibitory effects on cell proliferation. Taken together, our data indicate that apigenin downregulates Axl expression, which subsequently results in the inhibition of NSCLC cell proliferation through the increase and decrease of p21 and XIAP expression, respectively.


Improving selective androgen receptor modulator discovery and preclinical evaluation.
Selective androgen receptor modulators (SARMs) represent a new class of pharmaceuticals that may find wide clinical use. However, selectivity is not understood at the molecular level, which has made the discovery and preclinical evaluation of SARMs difficult. We review the current state of SARM discovery and preclinical evaluation, as well as our current understanding of the molecular mechanisms controlling AR selectivity. We then discuss how increasing our molecular knowledge of AR selectivity will help create better discovery and evaluation methods and lead to a wider array of safer SARMs. The SARM field has advanced rapidly, but without a solid foundation of molecular knowledge to inform discovery and preclinical evaluation methods. The field has also taken a narrow view of selectivity, disregarding many androgen-responsive tissues, which could lead to unforeseen and detrimental side effects with chronic administration of SARMs. An investment in basic research could accelerate the discovery of a new generation of more selective and safer SARMs that could be used to treat an expanded range of clinical conditions.


Three-year outcomes after acute kidney injury: results of a prospective parallel group cohort study.
Using a prospective study design, we aimed to characterise the effect of acute kidney injury (AKI) on long-term changes in renal function in a general hospital population. Hospitalised patients with AKI (exposed) and hospitalised patients without AKI (non-exposed), recruited at 3 months after hospital admission. Prospective, matched parallel group cohort study, in which renal function and proteinuria were measured at 3 months, 1 year and 3 years. Single UK centre. Clinical end points at 3 years were comparison of the following variables between exposed and non-exposed groups: renal function, prevalence of proteinuria and albuminuria and chronic kidney disease (CKD) progression/development at each time point. CKD progression was defined as a decrease in the estimated glomerular filtration rate (eGFR) of ≥25% associated with a decline in eGFR stage. 300 exposed and non-exposed patients were successfully matched 1:1 for age and baseline renal function; 70% of the exposed group had AKI stage 1. During follow-up, the AKI group had lower eGFR than non-exposed patients at each time point. At 3 years, the mean eGFR was 60.7±21 mL/min/1.73 m2 in the AKI group compared with 68.4±21 mL/min/1.73 m2 in the non-exposed group, p=0.003. CKD development or progression at 3 years occurred in 30 (24.6%) of the AKI group compared with 10 (7.5%) of the non-exposed group, p<0.001. Albuminuria was more common in the AKI group, and increased with AKI severity. Factors independently associated with CKD development/progression after AKI were non-recovery at 90 days, male gender, diabetes and recurrent AKI. AKI is associated with deterioration in renal function to 3 years, even in an unselected population with predominantly AKI stage 1. Non-recovery from AKI is an important factor determining long-term outcome.


Regeneration of stevia plant through callus culture.
Stevia rebaudiana Bertoni that conventionally propagated by seed or by cuttings or clump division which has a limitation of quality and quantity seed material. In present study, callus culture technique was tried to achieve rapid plant multiplication for quality seed material. Callus induction and multiplication medium was standardized from nodal as well as leaf sagments. It is possible to maintain callus on Murashige and Skoog medium supplemented with 6-benzyl amino purine and naphthalene acetic acid. Maximum callus induction was obtained on Murashige and Skoog medium incorporated with 6-benzyl amino purine (2.0-3.0 mg/l) and naphthalene acetic acid (2.0 mg/l) treatments. However, Murashige and Skoog medium containing 2.0 mg/l 6-benzyl amino purine+2.0 mg/l naphthalene acetic acid was found to be the best for callus induction. Higher regeneration frequency was noticed with Murashige and Skoog medium supplemented with 2.0 mg/l 6-benzyl amino purine+0.2 mg/l naphthalene acetic acid. Regenerated plants were rooted better on (1/4) Murashige and Skoog strength supplemented with 0.1 mg/l indole-3-butyric acid. The rooted plantlets were hardened successfully in tera care medium with 63 per cent survival rate. The developed protocol can be utilized for mass production of true to type planting material on large scale independent of season, i.e. external environmental conditions.


Predictors of readmission after outpatient plastic surgery.
Hospital readmissions have become a topic of focus for quality care measures and cost-reduction efforts. However, no comparative multi-institutional data on plastic surgery outpatient readmission rates currently exist. The authors endeavored to investigate hospital readmission rates and predictors of readmission following outpatient plastic surgery. The 2011 National Surgical Quality Improvement Program database was reviewed for all outpatient procedures. Unplanned readmission rates were calculated for all 10 tracked surgical specialties (i.e., general, thoracic, vascular, cardiac, orthopedics, otolaryngology, plastics, gynecology, urology, and neurosurgery). Multivariate logistic regression models were used to determine predictors of readmission for plastic surgery. A total of 7005 outpatient plastic surgery procedures were isolated. Outpatient plastic surgery had a low associated readmission rate (1.94 percent) compared with other specialties. Seventy-five patients were readmitted with a complication. Multivariate regression analysis revealed obesity (body mass index ≥ 30), wound infection within 30 days of the index surgery, and American Society of Anesthesiologists class 3 or 4 physical status as significant predictors for unplanned readmission. Unplanned readmission after outpatient plastic surgery is infrequent and compares favorably to rates of readmission among other specialties. Obesity, wound infection within 30 days of the index operation, and American Society of Anesthesiologists class 3 or 4 physical status are independent predictors of readmission. As procedures continue to transition into outpatient settings and the drive to improve patient care persists, these findings will serve to optimize outpatient surgery use.


Bridging the gap: health equality and the deficit framing of health.
The analyst tasked with measuring population health, with appraising healthcare investments, or allocating healthcare resources, may frame their task in one of two possible ways: either as being concerned with health assets (e.g. health expectancy or stock of QALYs), or with health deficits (a 'health gap', analogous to the poverty gap). In this paper, we discuss the consequences of taking the asset or the deficit concept as one's basic building block in developing a health measurement system when one has concerns about equitable distribution. We conclude that building metrics from a primitive health gap concept is possible and indeed may offer insights not otherwise easily accessible.


Sequential Linkage of Carbohydrate Antigens to Mimic Capsular Polysaccharides: Toward Semisynthetic Glycoconjugate Vaccine Candidates against Streptococcus pneumoniae Serotype 14.
Vaccines based on isolated polysaccharides successfully protect humans from bacterial pathogens such as Streptococcus pneumoniae. Because polysaccharide production and isolation can be technically challenging, glycoconjugates containing synthetic antigens are an attractive alternative. Typically, the shortest possible oligosaccharide antigen is preferable as syntheses of longer structures are more difficult and time-consuming. Combining several protective epitopes or polysaccharide repeating units as blocks by bonds other than glycosidic linkages would greatly reduce the synthetic effort if the immunological response to the polysaccharide could be retained. To explore this concept, we bridged the well-understood and immunologically potent RU of S. pneumoniae serotype 14 (ST14) with an aliphatic spacer and conjugated it to the carrier protein CRM197. Mice immunized with the spacer-bridged glycan conjugates produced high levels of specific antibodies after just one or two vaccine doses, while the tetrasaccharide repeating unit alone required three doses. The antibodies recognized specifically ST14 CPS, while no significant antibody levels were raised against the spacer or unrelated CPS. Synthetic vaccines generated antibodies with opsonic activity. Mimicking polysaccharides by coupling repeating unit antigens via an aliphatic spacer may prove useful also for the development of other glycoconjugate vaccine candidates, thereby reducing the synthetic complexity while enhancing a faster immune response.


Hyper-Acetylation of Histone H3K56 Limits Break-Induced Replication by Inhibiting Extensive Repair Synthesis.
Break-induced replication (BIR) has been implicated in restoring eroded telomeres and collapsed replication forks via single-ended invasion and extensive DNA synthesis on the recipient chromosome. Unlike other recombination subtypes, DNA synthesis in BIR likely relies heavily on mechanisms enabling efficient fork progression such as chromatin modification. Herein we report that deletion of HST3 and HST4, two redundant de-acetylases of histone H3 Lysine 56 (H3K56), inhibits BIR, sensitizes checkpoint deficient cells to deoxyribonucleotide triphosphate pool depletion, and elevates translocation-type gross chromosomal rearrangements (GCR). The basis for deficiency in BIR and gene conversion with long gap synthesis in hst3Δ hst4Δ cells can be traced to a defect in extensive DNA synthesis. Distinct from other cellular defects associated with deletion of HST3 and HST4 including thermo-sensitivity and elevated spontaneous mutagenesis, the BIR defect in hst3Δ hst4Δ cannot be offset by the deletion of RAD17 or MMS22, but rather by the loss of RTT109 or ASF1, or in combination with the H3K56R mutation, which also restores tolerance to replication stress in mrc1 mutants. Our studies suggest that acetylation of H3K56 limits extensive repair synthesis and interferes with efficient fork progression in BIR.


When thought suppression backfires: its moderator effect on eating psychopathology.
Recently, several studies have pointed the importance of thought suppression as a form of experiential avoidance in different psychopathological conditions. Thought suppression may be conceptualized as an attempt to decrease or eliminate unwanted internal experiences. However, it encloses a paradoxical nature, making those thoughts hyper accessible and placing an extra burden on individuals. This avoidance process has been associated with several psychopathological conditions. However, its role in eating psychopathology remains unclear. The present study aims to explore the moderation effect of thought suppression on the associations between body image-related unwanted internal experiences (unfavorable social comparison through physical appearance and body image dissatisfaction) and eating psychopathology severity in a sample of 211 female students. Correlational analyses showed that thought suppression is associated with psychological inflexibility and eating disorders' main risk factors and symptoms. Moreover, two independent analyses revealed that thought suppression moderates, as it amplifies, the impact of unfavorable social comparisons through physical appearance (model 1) and body image dissatisfaction (model 2) on disordered eating attitudes and behaviors. Hence, for the same level of these body-related internal experiences, young females who reveal higher levels of thought suppression present higher eating psychopathology. Taken together, these findings highlight the key role of thought suppression in eating psychopathology and present important clinical implications.


Metabolites of N-ethylbenzamide, N,N-diethylbenzamide and related compounds detected in rat urine by NMR spectroscopy.
In rats treated ip with N-ethylbenzamide (EB) at doses of 250-500 mg/kg, this amide was metabolized by hydrolysis to ethylamine and benzoic acid, the latter detected in the urine as hippuric acid. Proposed metabolic pathways were investigated by treatment of male Wistar rats with EB and N-ethyl-alpha-13C-benzamide followed by examination of urine samples by high-resolution NMR spectroscopy. Ethylamine was detected by 1H and 13C NMR techniques. Estimation of metabolite levels in 24-hr posttreatment urine by 1H NMR spectroscopy with an internal standard of 3-(trimethylsilyl)-1-propanesulfonic acid and correction for background levels of hippuric acid in control urine showed that the percentage of the dose excreted as hippuric acid was 52-76%. The corresponding range for the excretion of ethylamine (detected as an ethylammonium cation) was 27-41%. Signals for EB, benzamide, and benzoic acid were not above the background in urine of treated animals. A similar NMR study with unlabeled p-chloro-N-ethylbenzamide (p-Cl EB) resulted in the detection of p-chlorohippuric acid and ethylamine. Treatment of rats with N,N-diethylbenzamide and p-chloro-N,N-diethylbenzamide gave the same detectable metabolites in the urine as those found in experiments with EB and p-Cl EB. These observations could be rationalized with a metabolic pathway involving an initial oxidative mono-N-deethylation reaction followed by enzymatic hydrolysis of the secondary amides to ethylamine and benzoic acids and conjugation of the latter with glycine. N-Methylbenzamide was also found to be eliminated in the urine as hippuric acid.


Recent nutrition research: implications for foods of the future.
Dietary habits and the foods we ingest influence health. Nutrition is one factor that can be controlled in such a way as to influence an individual's destiny in such diseases as coronary heart disease (CHD) hypertension, stroke, cancer, osteoporosis, diabetes and gall stone disease. The role of nutrition in treatment and prevention of CHD is one example. It is not only a problem in Western Europe and in the United States anymore but with "westernization" of Asia and Africa and in many Eastern European countries, CHD is increasing at an alarming rate. Led by the American Heart Association, influential groups have provided dietary recommendations to the general public over the past 30 years. These recommendations have been based to a large extent on research carried out to determine the influence of various dietary components, particularly fatty acids and dietary cholesterol, on the risk factors leading to CHD. The results of these investigations can now be used by industry to provide foods for the future which will provide the public with more healthy choices and hopefully aid in the control of diseases which can be influenced by diet.


Innervation of the monkey vas deferens.
Fluorescent and acetylcholinesterase-positive cell bodies occur in the connective tissue surrounding the prostatic end of the monkey vas deferens. At both the testicular and prostatic ends, noradrenergic nerve fibres are distributed mainly in the smooth muscle coats while acetylcholinesterase-positive fibres are most densely distributed in the lamina propria but also quite significantly in the inner part of the smooth musculature. The percentage of cholinergic varicosities (containing a predominance of small agranular vesicles and some large granular vesicles) is higher in the smooth muscle coat than in the lamina propria at both the testicular (9.1% against 1.5%) and prostatic (19.38% against 5.16%) ends of the vas deferens and the percentages of cholinergic varicosities in both the lamina propria and smooth muscle coat (5.16% and 19.38% respectively) at the prostatic end are higher than those (1.5% and 9.1% respectively) at the testicular end. The percentage of noradrenergic varicosities (containing a predominance of small granular vesicles interspersed with some large granular vesicles and agranular vesicles) in the muscle coat is higher at the testicular (56.72%) than the prostatic (39.73%) end. In addition to cholinergic and noradrenergic nerves, varicosities resembling purinergic nerve fibres, varicosities containing flattened vesicles and sensory terminals are also present. In nearly all cases where no collagen fibres intervene between an axon varicosity and the smooth muscle membrane, the interval between the two is less than 180 nm, and may be as little as, or even less than, 20 nm. Lastly, close nerve-mast cell and nerve-fibroblast contacts have been observed.


Adrenergic pharmacology of human and canine peripheral veins.
A comparison has been made of the factors concerned with the response of canine and human saphenous veins to adrenergic stimulation. Both vessels have prejunctional muscarinic and beta-adrenergic receptors. When activated by appropriate agonists these receptors decrease and increase the output, respectively, of norepinephrine from the nerve endings. Both vessels have postjunctional alpha 1 and alpha 2 adrenoceptors and postjunctional beta adrenoceptors. Activation of the former two receptors leads to contraction of the smooth muscle, and of the latter to relaxation. There are, however, qualitative differences. In the human veins the responsiveness of the prejunctional beta adrenoceptors exceeds that of the postjunctional, whereas the reverse is true in the dog. As a consequence, in the human vein beta-adrenergic agonists augment, and in the canine veins they depress, the contractile response to sympathetic nerve stimulation.


Reduced kidney transplant rejection rate and pharmacoeconomic advantage of mycophenolate mofetil.
Several multinational controlled clinical trials have shown that triple therapy immunosuppressive regimens which include mycophenolate mofetil (MMF), cyclosporin A (CSA) and steroids (S) are superior compared with conventional regimens which include azathioprine (AZA), CSA and S, mainly because MMF reduces the rate of acute rejection episodes in the first 6 months after kidney transplantation. Post-marketing studies are useful to evaluate the general applicability and costs of MMF-based immunosuppressive regimens. Based on the excellent results of the published controlled clinical trials, we have changed the standard triple therapy immunosuppressive protocol (AZA+CSA+S) to an MMF-based regimen (MMF+CSA+S) at our centre. To analyse the impact of this change in regimen, we have monitored 6-month patient and graft survival, rejection rate, serum creatinine and CSA levels, as well as the costs of the immunosuppressive and anti-rejection treatments, in 40 consecutive renal transplant recipients (MMF group) and have compared the data with 40 consecutive patients transplanted immediately prior to the change in regimen (AZA group). Recipient and donor characteristics were similar in the AZA and MMF groups. Patient survival (37/40; 92.5% in the AZA group vs 38/40; 95% in the MMF group), graft survival (36/40 vs 36/40; both 90%) and serum creatinine (137+/-56 vs 139+/-44 micromol/l) after 6 months were not significantly different. However, the rate of acute rejection episodes (defined as a rise in creatinine without other obvious cause and treated at least with pulse steroids) was significantly reduced with MMF from 60 to 20% (P=0.0005). The resulting cost for rejection treatment was lowered 8-fold (from sFr. 2113 to 259 averaged per patient) and the number of transplant biopsies was lowered > 3-fold in the MMF group. The cost for the immunosuppressive therapy was increased 1.5-fold with MMF (from sFr. 5906 to 9231 per patient for the first 6 months). The change from AZA to MMF resulted in a significant reduction in early rejection episodes, resulting in fewer diagnostic procedures and rehospitalizations. The optimal long-term regimen in terms of patient and pharmacoeconomic benefits remains to be defined.


Potentiating SLC transporter activity: Emerging drug discovery opportunities.
Maintaining the integrity of cellular membranes is critical to protecting metabolic activities and genetic information from the environment. Regulation of transport across membranes of essential chemicals, including water, nutrients, hormones and many drugs, is therefore key to cellular homeostasis and physiological processes. The two main transporter superfamilies are ATP-binding cassette (ABC) transporters that primarily function as efflux transporters, and the solute carrier (SLC) transporters. SLC transporters encompass 52 gene families with almost 400 different human transporter genes. Although long under-explored, SLC transporters are an emerging drug target class and the molecular target of several approved inhibitor drugs, such as selective serotonin reuptake inhibitors (SSRIs) for depression and sodium/glucose co-transporter (SGLT2) inhibitors for diabetes. Interestingly though, although loss-of-function mutations in numerous human SLC transporters are linked to Mendelian diseases, few reports of SLC transporter activators have appeared, and only inhibitors have been advanced to clinical studies. In this commentary, we discuss several strategies for potentiating SLC transporter function, from direct acting potentiators to modulators of transcription, translation or trafficking. We review the progress made in recent years toward the understanding of the structural and molecular basis of SLC transporter function and the pathways and mechanisms that regulate SLC expression, and describe the opportunities these new insights present for discovery of SLC transporter potentiators. Finally, we highlight the challenges associated with the various approaches and provide some thoughts on future directions that might facilitate the search for SLC potentiators with therapeutic potential.


First-phase insulin secretion: does it exist in real life? Considerations on shape and function.
To fulfill its preeminent function of regulating glucose metabolism, insulin secretion must not only be quantitatively appropriate but also have qualitative, dynamic properties that optimize insulin action on target tissues. This review focuses on the importance of the first-phase insulin secretion to glucose metabolism and attempts to illustrate the relationships between the first-phase insulin response to an intravenous glucose challenge and the early insulin response following glucose ingestion. A clear-cut first phase occurs only when the beta-cell is exposed to a rapidly changing glucose stimulus, like the one induced by a brisk intravenous glucose administration. In contrast, peripheral insulin concentration following glucose ingestion does not bear any clear sign of biphasic shape. Coupling data from the literature with the results of a beta-cell model simulation, a close relationship between the first-phase insulin response to intravenous glucose and the early insulin response to glucose ingestion emerges. It appears that the same ability of the beta-cell to produce a pronounced first phase in response to an intravenous glucose challenge can generate a rapidly increasing early phase in response to the blood glucose profile following glucose ingestion. This early insulin response to glucose is enhanced by the concomitant action of incretins and neural responses to nutrient ingestion. Thus, under physiological circumstances, the key feature of the early insulin response seems to be the ability to generate a rapidly increasing insulin profile. This notion is corroborated by recent experimental evidence that the early insulin response, when assessed at the portal level with a frequent sampling, displays a pulsatile nature. Thus, even though the classical first phase does not exist under physiological conditions, the oscillatory behavior identified at the portal level does serve the purpose of rapidly exposing the liver to elevated insulin levels that, also in virtue of their up-and-down pattern, are particularly effective in restraining hepatic glucose production.


An insect TEP in a crustacean is specific for cuticular tissues and involved in intestinal defense.
In an attempt to identify genes encoding thioester-containing proteins in the freshwater crayfish, Pacifastacus leniusculus, three different cDNAs were found. A phylogenetic analysis of these proteins indicates that they can be classified into two subfamilies: two alpha-2-macroglobulins (Pl-A2M1, Pl-A2M2) showing a close similarity to shrimp A2M, and one insect TEP-like protein (Pl-TEP). This is the first report of an insect TEP-like protein in a crustacean. Crayfish Pl-A2M1, Pl-A2M2 and Pl-TEP cDNAs encode proteins with 1480, 1586 or 1507 amino acids, respectively. Pl-A2M1, Pl-A2M2 and Pl-TEP have the basic domain structure and functionally important residues for each molecule, and their mRNA was detected in different parts of the body, suggesting that they may have different functions. Pl-A2M1 was mainly expressed in hemocytes and Pl-A2M2 was highly expressed in heart and nerve, while Pl-TEP was exclusively expressed in cuticular tissues such as gill and intestine. RNA interference of Pl-TEP in vivo resulted in that these animals were slightly less resistant when fed with the bacterium, Pseudomonas libanensis/gessardii. Furthermore, when TEP activity was blocked using methylamine followed by bacterial feeding, the animals were killed to a higher extent compared to a control group. Taken together, this indicates that Pl-TEP and/or Pl-A2M1, Pl-A2M2 may be important for the immune defense in crayfish intestine and function as a pattern recognition protein in crayfish cuticular tissues.


Expression and cellular localization of inhibin alpha-subunit mRNA in equine fetal gonads.
The expression of inhibin alpha-subunit mRNA in equine fetal gonads during pregnancy (Days 90 to 300) was examined by means of Northern blot analysis. In all samples examined, a single species of transcript was detected at the size of 1.5 kb. A digoxigenin-labeled antisense cRNA probe specific to equine inhibin alpha-subunit was synthesized and in situ hybridization analysis to locate the inhibin alpha-subunit mRNA positive cells was performed using frozen tissue sections of equine fetal ovary (day 150 of pregnancy) and equine fetal testis (day 180 of pregnancy). In the fetal ovary, positive cells were seen throughout the interstitial area but did not show any particular localization. In the fetal testis, on the other hand, the antisense cRNA hybridized almost exclusively to the interstitial cells surrounding developing seminiferous cords and Sertoli cells within the cords. Positive signals were also detected in a limited number of the interstitial cells located away from the cords. These results suggest that in equine fetal gonads, inhibin and/or inhibin alpha-subunit related molecules such as the monomeric form are produced and these molecules may have a paracrine/autocrine role within the gonads.


Mood and memory deficits in a model of Gulf War illness are linked with reduced neurogenesis, partial neuron loss, and mild inflammation in the hippocampus.
Impairments in mood and cognitive function are the key brain abnormalities observed in Gulf war illness (GWI), a chronic multisymptom health problem afflicting ∼25% of veterans who served in the Persian Gulf War-1. Although the precise cause of GWI is still unknown, combined exposure to a nerve gas prophylaxis drug pyridostigmine bromide (PB) and pesticides DEET and permethrin during the war has been proposed as one of the foremost causes of GWI. We investigated the effect of 4 weeks of exposure to Gulf war illness-related (GWIR) chemicals in the absence or presence of mild stress on mood and cognitive function, dentate gyrus neurogenesis, and neurons, microglia, and astrocytes in the hippocampus. Combined exposure to low doses of GWIR chemicals PB, DEET, and permethrin induced depressive- and anxiety-like behavior and spatial learning and memory dysfunction. Application of mild stress in the period of exposure to chemicals exacerbated the extent of mood and cognitive dysfunction. Furthermore, these behavioral impairments were associated with reduced hippocampal volume and multiple cellular alterations such as chronic reductions in neural stem cell activity and neurogenesis, partial loss of principal neurons, and mild inflammation comprising sporadic occurrence of activated microglia and significant hypertrophy of astrocytes. The results show the first evidence of an association between mood and cognitive dysfunction and hippocampal pathology epitomized by decreased neurogenesis, partial loss of principal neurons, and mild inflammation in a model of GWI. Hence, treatment strategies that are efficacious for enhancing neurogenesis and suppressing inflammation may be helpful for alleviation of mood and cognitive dysfunction observed in GWI.


[Practical aspects of partition measurements according to GLP rules].
Experimental methods for octanol/water partition coefficient (logP) determination are surveyed. The terminology used in the literature, the lipophilicity/pH profile and the most important factors influencing the logP values have been discussed. Several new, recently developed direct logP determination methods are introduced including their advantages and limits of application. Some aspects of good laboratory practice of the shake-flask method are described and results of a validation study of pH-metric logP determination technique used PCA 101 pKa and logP analyser (Sirius, UK) are also shown. Questions have to be answered in method selection for logP measurement are summarized in flow chart (Fig. 8). The author based on her own experiences in lipophilicity measurements over decades, suggests the shake-flask method and the automated dual-phase potentiometric technique, as approaches fulfilling the GLP rules.


Evidence for direct geographic influences on linguistic sounds: the case of ejectives.
We present evidence that the geographic context in which a language is spoken may directly impact its phonological form. We examined the geographic coordinates and elevations of 567 language locations represented in a worldwide phonetic database. Languages with phonemic ejective consonants were found to occur closer to inhabitable regions of high elevation, when contrasted to languages without this class of sounds. In addition, the mean and median elevations of the locations of languages with ejectives were found to be comparatively high. The patterns uncovered surface on all major world landmasses, and are not the result of the influence of particular language families. They reflect a significant and positive worldwide correlation between elevation and the likelihood that a language employs ejective phonemes. In addition to documenting this correlation in detail, we offer two plausible motivations for its existence. We suggest that ejective sounds might be facilitated at higher elevations due to the associated decrease in ambient air pressure, which reduces the physiological effort required for the compression of air in the pharyngeal cavity--a unique articulatory component of ejective sounds. In addition, we hypothesize that ejective sounds may help to mitigate rates of water vapor loss through exhaled air. These explications demonstrate how a reduction of ambient air density could promote the usage of ejective phonemes in a given language. Our results reveal the direct influence of a geographic factor on the basic sound inventories of human languages.