Social exclusion reliably engages the default network: A meta-analysis of Cyberball.
Social exclusion refers to the experience of being disregarded or rejected by others and has wide-ranging negative consequences for well-being and cognition. Cyberball, a game where a ball is virtually tossed between players, then leads to the exclusion of the research participant, is a common method used to examine the experience of social exclusion. The neural correlates of social exclusion remain a topic of debate, particularly with regards to the role of the dorsal anterior cingulate cortex (dACC) and the concept of social pain. Here we conducted a quantitative meta-analysis using activation likelihood estimation (ALE) to identify brain activity reliably engaged by social exclusion during Cyberball task performance (Studies = 53; total N = 1,817 participants). Results revealed consistent recruitment in ventral anterior cingulate and posterior cingulate cortex, inferior and superior frontal gyri, posterior insula, and occipital pole. No reliable activity was observed in dACC. Using a probabilistic atlas to define dACC, fewer than 15% of studies reported peak coordinates in dACC. Meta-analytic connectivity mapping suggests patterns of co-activation are consistent with the topography of the default network. Reverse inference for cognition associated with reliable Cyberball activity computed in Neurosynth revealed social exclusion to be associated with cognitive terms Social, Autobiographical, Mental States, and Theory of Mind. Taken together, these findings highlight the role of the default network in social exclusion and warns against interpretations of the dACC as a key region involved in the experience of social exclusion in humans.


Body composition, lipid profile and clinical parameters are predictors of prognosis in patients with heart failure: Two-year follow-up.
Heart failure (HF) is a complex syndrome that leads to changes in body composition and eventually results in unfavorable outcomes. This study aimed to evaluate body composition, lipid profiles and clinical parameters of patients with HF, and their associations with both survival and unfavorable clinical outcomes. This prospective cohort study included 94 adults and older people with HF. Body composition was assessed by bioelectrical impedance analysis (BIA). Anthropometric variables and lipid profile were also evaluated. Electronic medical records were checked to collect information on clinical outcomes (mortality and hospitalization), considering a follow-up period of 24 months. Survival was calculated using the Kaplan-Meier estimate, and the curves compared using Log-Rank. The death risk rate (Hazard Ratio, HR) was calculated using Cox's univariate models. Mean age was 55.1 (13.9) years and there was a higher frequency of males. There was a predominance of HF with reduced ejection fraction, and ischemic etiology. Patients with New York Heart Association (NYHA) functional classification I/II had a better overall survival rate at 24 months than those with NYHA III/IV (univariate HR 4.93 (1.76-13.82); p = 0.001). Greater survival rates were found in patients without chronic kidney disease (CKD) (univariate HR 2.93 (1.59-5.39); p = 0.01). In the multivariate analyses, both dyslipidemia (adjusted HR 3.84 (1.22-12.00); p = 0.021) and increased fat mass index (FMI) were associated with overall survival rate (adjusted HR 3.59 (1, 10-11.74); p = 0.034). The severity of HF symptoms and the presence of chronic kidney disease are associated with higher mortality. Increased fat mass index and dyslipidemia are predictors of favorable outcomes in this population.


Briefing in family characteristics of microRNAs and their applications in cancer research.
MicroRNAs (miRNAs) are endogenous, short, non-coding RNA molecules that are directly involved in the post-transcriptional regulation of gene expression. Dysregulation of miRNAs is usually associated with diseases. Since miRNAs in a family intend to have common functional characteristics, proper assignment of miRNA family becomes heuristic for better understanding of miRNA nature and their potentials in clinic. In this review, we will briefly discuss the recent progress in miRNA research, particularly its impact on protein and its clinical application in cancer research in a view of miRNA family. This article is part of a Special Issue entitled: Computational Proteomics, Systems Biology & Clinical Implications. Guest Editor: Yudong Cai.


Fluoxetine in the environment may interfere with the neurotransmission or endocrine systems of aquatic animals.
Antidepressants are extensively used to treat the symptoms of depression in humans, and the environmentally discharged drugs potentially threaten aquatic organisms. In this study, the acute toxic effects of fluoxetine (FLX) were investigated in two aquatic organisms, the freshwater polyp (Hydra magnipapillata) and Javanese medaka (Oryzias javanicus). The median lethal concentration (LC50) of FLX in H. magnipapillata was 3.678, 3.082, and 2.901 mg/L after 24, 48, and 72 h, respectively. Morphological observations of the FLX-exposed H. magnipapillata showed that 1.5 mg/L FLX induced the contraction of the tentacles and body column. The LC50 of FLX in O. javanicus was 2.046, 1.936, 1.532, and 1.237 mg/L after 24, 48, 72, and 96 h, respectively. Observation of the behavior of the FLX-exposed fish showed that FLX reduced their swimming performance at a minimum concentration of 10 µg/L. The half-maximal effective concentration (EC50) of FLX for swimming behavior in O. javanicus was 0.135, 0.108, and 0.011 mg/L after 12, 24, and 96 h, respectively. Transcriptomic analyses indicated that FLX affects various physiological and metabolic processes in both species. FLX exposure induced oxidative stress, reproductive deficiency, abnormal pattern formation, DNA damage, and neurotransmission disturbance in H. magnipapillata, whereas it adversely affected O. javanicus by inducing oxidative stress, DNA damage, endoplasmic reticulum stress, and mRNA instability. Neurotransmission-based behavioral changes and endocrine disruption were strongly suspected in the FLX-exposed fish. These results suggest that FLX affects the behavior and metabolic regulation of aquatic organisms.


Differentiated thyroid cancer: a stage adapted approach to the treatment of regional lymph node metastases.
The controversy in the management of regional lymph nodes in patients with differentiated thyroid cancer is discussed on the basis of a review of the literature. Since no prospective studies have yet compared limited dissections ('node picking') with more extensive dissections [(modified) radical neck dissection], a retrospective analysis was performed using two patient groups in which patients were managed differently with regard to the preoperative diagnosis and treatment of regional lymph node metastases. Only patients with proven lymph node metastases were included in the study. Because of selection methods necessary to create comparable patient groups, only 83 patients could be included in the analysis. There was no difference in survival or recurrence rate in either group, although recurrences occurred less frequently in the explored side of the neck after MRND (3.9% vs. 6.3%). More postoperative morbidity was found in the patients who had been subjected to a more extensive search for and treatment of lymph node metastases. Because of the relatively small number of patients only the difference in occurrence of accessory nerve palsies reached statistical significance (P = 0.05). It is advocated that only in the case of papillary carcinoma with limited lymph node involvement node picking is the procedure of choice. In all other cases a modified radical neck dissection should be standard treatment.


Quantification of 11 enzyme activities of lysosomal storage disorders using liquid chromatography-tandem mass spectrometry.
Lysosomal storage disorders (LSDs) are characterized by the accumulation of lipids, glycolipids, oligosaccharides, mucopolysaccharides, and other biological substances because of the pathogenic deficiency of lysosomal enzymes. Such diseases are rare; thus, a multiplex assay for these disorders is effective for the identification of affected individuals during the presymptomatic period. Previous studies have demonstrated that such assays can be performed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) with multiple reaction monitoring (MRM) detection. An assay procedure to quantify the activity of 11 enzymes associated with LSDs was provided. First, a validation study was performed using dried blood spot (DBS) samples with 100% and 5% enzyme activity for quality control (QC). Under the assay condition, the analytical range, defined as the ratio of the peak area of the enzyme reaction products from the DBS for QC with 100% enzyme activity to that from the filter paper blank sample, was between 14 for GALN and 4561 for GLA. Based on these results, the distribution of the enzyme activity for the 11 LSD enzymes was further examined. Consistent with the previous data, the enzyme activity exhibited a bell-shaped distribution with a single peak. The averaged enzyme activity for the healthy neonates was as follows: GLA, 3.80 ± 1.6; GAA, 10.6 ± 4.8; IDUA, 6.4 ± 2.3; ABG, 8.6 ± 3.1; ASM, 3.3 ± 1.1; GALC, 2.8 ± 1.3; ID2S, 16.7 ± 6.1; GALN, 1.2 ± 0.5; ARSB, 17.0 ± 8.7; NAGLU, 4.6 ± 1.5; and GUSB, 46.6 ± 19.0 μmol/h/L (mean ± SD, n = 200). In contrast, the enzyme activity in disease-affected individuals was lower than the minimum enzyme activity in healthy neonates. The results demonstrate that the population of disease-affected individuals was distinguished from that of healthy individuals by the use of LC-MS/MS.


Identification and characterization of a motilin-like peptide and its receptor in teleost.
Although putative motilin receptor sequences have been reported in teleost, there is no proof for the existence of the motilin gene in teleost. In this study, we have identified a motilin-like gene in the genome of several fish species and cloned its cDNA sequence from zebrafish. The zebrafish motilin-like precursor shares very low amino acid (aa) identities with the previously reported motilin precursors. Processing of the zebrafish motilin-like precursor may generate a 17-aa C-terminal amidated mature peptide, the motilin-like peptide (motilin-LP). A putative zebrafish motilin receptor (MLNR) was also identified in zebrafish. In cultured eukaryotic cells transfected with the zebrafish MLNR, zebrafish motilin-LP could enhance both CRE-driven and SRE-driven promoter activities. Tissue distribution studies indicated that the zebrafish motilin-like gene is mainly expressed in the intestine and liver while the zebrafish MLNR gene is highly expressed in brain regions, suggesting that motilin-LP behaves like other gut hormones to regulate brain functions. These data suggest that the presence of a unique motilin/MNLR system in teleost.


FIA acid-base titrations with a new flow-through pH detector.
A pH-sensitive detector for flow-through potentiometry based on a graphite/quinhydrone composite electrode was applied for flow-injection analysis (FIA) titrations. Hydrochloric acid and acetic acid were titrated by injection of samples into a sodium hydroxide carrier solution. System conditions were optimised by variation of flow rate, injection volume and titrant concentration. The parameters sampling frequency, residence time and dispersion coefficient were determined. The evaluation of peak width (time between the two inflection points on each side of the peak), peak area and slope of the raising edge of the peak lead were studied with respect to their use for calibration. Hydrochloric acid and acetic acid could be titrated down to a concentration of 2 x 10(-4) mol L(-1) using 150-microL injection volumes, which is almost ten times lower than can be achieved using colour indicators and a spectrophotometric detection.


Review: pathophysiology of intracranial hypertension and noninvasive intracranial pressure monitoring.
Measurement of intracranial pressure (ICP) is crucial in the management of many neurological conditions. However, due to the invasiveness, high cost, and required expertise of available ICP monitoring techniques, many patients who could benefit from ICP monitoring do not receive it. As a result, there has been a substantial effort to explore and develop novel noninvasive ICP monitoring techniques to improve the overall clinical care of patients who may be suffering from ICP disorders. This review attempts to summarize the general pathophysiology of ICP, discuss the importance and current state of ICP monitoring, and describe the many methods that have been proposed for noninvasive ICP monitoring. These noninvasive methods can be broken down into four major categories: fluid dynamic, otic, ophthalmic, and electrophysiologic. Each category is discussed in detail along with its associated techniques and their advantages, disadvantages, and reported accuracy. A particular emphasis in this review will be dedicated to methods based on the use of transcranial Doppler ultrasound. At present, it appears that the available noninvasive methods are either not sufficiently accurate, reliable, or robust enough for widespread clinical adoption or require additional independent validation. However, several methods appear promising and through additional study and clinical validation, could eventually make their way into clinical practice.


A Novel Composite Metric for Predicting Patient Satisfaction With Spinal Cord Stimulation.
To identify relationships between clinical assessments of chronic pain to enable the generation of a multivariate model to predict patient satisfaction with spinal cord stimulation (SCS) treatment. Data from an exploratory clinical trial of sub-perception SCS (SPSCS) were reviewed. Forty-seven subjects tested multiple SPSCS programs for three to four days each. At the end of each program period, subjects recorded pain intensity, patient satisfaction with treatment (PSWT), modified patient global impression of change, and physical activity tolerance times. Twelve outcome variables were evaluated. Pearson's correlation coefficient was used to assess pair-wise correlations. Multigenerational mixed effects modeling was performed to create a model to best explain relationships between those variables. A final model was generated that predicted PSWT using evening pain intensity (EPI) and the interaction between EPI and walking tolerance time. The mixed effects model allows for visualization of the interactions between EPI, walking tolerance time, and patient satisfaction with SCS. Patient-centered outcomes are desirable when evaluating complex multidimensional health impairments but accurately predicting patient satisfaction with treatment remains a challenge. Understanding the variables that predict (either by causation or association) satisfaction would be useful for clinicians. The results of this study suggest that a composite measure of activity tolerance (i.e., walking tolerance) and pain intensity can predict patient satisfaction with SCS therapy. This study highlights the utility of composite outcomes metrics in evaluating the benefits of SCS for chronic low back and leg pain.


Prognostic importance of RASSF2 expression in patients with gastric cancer who had undergone radical gastrectomy.
Although Ras-association domain family of gene 2 (RASSF2) has been shown to undergo promoter methylation at high frequency in some cancer types and in brain metastases, its clinical utility as a useful prognostic molecular marker remains unclear in gastric cancer. Prognostic significance of RASSF2 expression was retrospectively analysed by immunohistochemically in 105 patients with gastric cancer who underwent curative gastrectomy. Low RASSF2 expression was detected in 58 (55 %) patients, whereas 47 patients (45 %) had high RASSF2 expression. Lymph node involvement, pT stage, TNM stage, vascular invasion, perineural invasion and the presence of recurrence were found to be significantly related to RASSF2 expression levels. Low PRL-3 expression was closely correlated with lymph node metastasis (p = 0.001), advanced pT stage (p = 0.021), advanced TNM stage (p < 0.001), the presence of vascular invasion (p < 0.001), perineural invasion (p = 0.018) and high prevalence of recurrence (p = 0.003) compared with high RASSF2 expression. The median disease-free survival (DFS) time for patients with low RASSF2 expression was significantly worse than that of patients with high RASSF2 expression (10.2 vs. 50.6 months, p < 0.001). In addition, patients with high RASSF2 expression had the higher overall survival (OS) interval compared to patients with low RASSF2 expression (NR vs. 14.9 months, p < 0.001). In the multivariate analysis, the rate of RASSF2 expression levels was an independent prognostic factor, for DFS [p < 0.001, HR 0.12 (0.10-0.88)] and OS [p < 0.001, HR 0.10 (0.04-0.46)], as were pT stage and TNM stage, respectively. RASSF2 may be an important molecular marker for carcinogenesis, prognosis and progression in gastric cancer, but the potential value of RASSF2 expression as a useful molecular marker in gastric cancer progression should be evaluated, comprehensively. It would be possible to develop treatments targeting RASSF2 and advance new treatment strategies for gastric cancer.


Reactive lymphoid hyperplasia 1 month after LASIK surgery.
This study was conducted to investigate a case of reactive lymphoid hyperplasia following laser assisted in situ keratomileusis (LASIK). A 31-year-old man who underwent LASIK presented 1 month later with a fleshy conjunctival (plical) tumor in the left eye. An excision biopsy of the tumor was performed. Histopathology of the excised tumor revealed reactive lymphoid hyperplasia. Conjunctival lymphomas can masquerade as chronic conjunctivitis and can be preceded by reactive lymphoid hyperplasia. It is important to identify and differentiate these tumors. This report describes the unusual occurrence of a lymphoid conjunctival tumor after LASIK eye surgery.


Non-metabolic role of alpha-enolase in virus replication.
Viruses are extremely complex and highly evolving microorganisms; thus, it is difficult to analyse them in detail. The virion is believed to contain all the essential components required from its entry to the establishment of a successful infection in a susceptible host cell. Hence, the virion composition is the principal source for its transmissibility and immunogenicity. A virus is completely dependent on a host cell for its replication and progeny production. Occasionally, they recruit and package host proteins into mature virion. These incorporated host proteins are believed to play crucial roles in the subsequent infection, although the significance and the molecular mechanism regulated are poorly understood. One such host protein which is hijacked by several viruses is the glycolytic enzyme, Enolase (Eno-1) and is also packaged into mature virion of several viruses. This enzyme exhibits a highly flexible nature of functions, ranging from metabolic to several non-metabolic activities. All the glycolytic enzymes are known to be moonlighting proteins including enolase. The non-metabolic functions of this moonlighting protein are also highly diverse with respect to its cellular localization. Although very little is known about the virological significance of this enzyme, several of its non-metabolic functions have been observed to influence the virus replication cycle in infected cells. In this review, we have attempted to provide a comprehensive picture of the non-metabolic role of Eno-1, its significance in the virus replication cycle and to stimulate interest around its scope as a therapeutic target for treating viral pathologies.


Prenatal Carbamazepine Exposure and Academic Performance in Adolescents: A Population-Based Cohort Study.
To investigate whether children born to mothers who used carbamazepine during pregnancy had worse academic performance in adolescence. This population-based cohort study included all live-born singletons in Denmark between 1996 and 2002 who participated in the national ninth-grade exit examination (n = 370,859). Those born to mothers with prescription of antiseizure medications other than carbamazepine during pregnancy were excluded. We examined the association of in utero exposure to maternal carbamazepine redeemed during pregnancy (n = 290) with academic performance of offspring, defined by the scores in Danish and mathematics in ninth-grade exit examination. We estimated mean z-score difference with linear regression adjusted for socioeconomic factors and potential indications, including epilepsy and medication for other psychiatric disorders. Additional analyses addressing confounding by indication included comparison between in utero exposed vs past exposed and between past exposed and never exposed. In utero exposure to valproate monotherapy was used as a positive control and in utero exposure to lamotrigine as a negative control. At the age of 16.1 (SD 0.4) years, adolescents in utero exposed to maternal carbamazepine monotherapy had lower scores both in Danish and mathematics in ninth-grade exit examination (adjusted z-score difference, -0.14 [95% CI -0.24 to -0.05] and -0.17 [95% CI -0.28 to -0.07], respectively). In utero exposure to carbamazepine monotherapy was associated with lower scores than past exposure only (adjusted z-score difference, -0.24 [95% CI -0.41 to -0.06] for Danish and -0.25 [95% CI -0.44 to -0.06] for mathematics), while past exposure to carbamazepine was associated with minor decrease in offspring's academic performance (adjusted z-score difference, -0.02 [95% CI -0.09 to 0.06] for Danish and -0.07 [95% CI -0.16 to 0.01] for mathematics). The association was also observed for in utero exposure to valproate monotherapy, but not for in utero exposure to lamotrigine. In utero exposure to carbamazepine was associated with poorer academic performance in adolescence, as represented by lower scores in ninth-grade exit examination in Danish and mathematics. Additional studies are needed to confirm these findings because of limitations in this study and variable findings in prior studies. This study provides Class III evidence that academic performance, as reflected in ninth-grade exit examinations in Danish and mathematics, was worse among those exposed to carbamazepine monotherapy in utero, compared with those without in utero exposure to antiseizure medications.


Effects of COVID-19 vaccination on platelet counts and bleeding in children, adolescents, and young adults with immune thrombocytopenia.
Coronavirus disease 2019 (COVID-19) vaccines rarely cause de novo immune thrombocytopenia (ITP) but may worsen preexisting ITP in adults. Whether severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccines impact platelet counts and bleeding in children, adolescents, and young adults (C-AYA) with preexisting ITP is unknown. We report here the very limited effect of COVID-19 vaccination on platelet counts and bleeding in a single-center series of 2 C-AYA with ITP. No patient experienced worsening bleeding and only one child had a significant decrease in platelet count which improved spontaneously to her baseline without intervention. SARS-CoV2 vaccination was safe in C-AYA with ITP in this small cohort.


Interactions of oxidative stress with thiamine homeostasis promote neurodegeneration.
Thiamine-dependent processes are diminished in brains of patients with several neurodegenerative diseases. The decline in thiamine-dependent enzymes can be readily linked to the symptoms and pathology of the disorders. Why the reductions in thiamine linked processes occur is an important experimental and clinical question. Oxidative stress (i.e. abnormal metabolism of free radicals) accompanies neurodegeneration and causes abnormalities in thiamine-dependent processes. The vulnerability of thiamine homeostasis to oxidative stress may explain deficits in thiamine homeostasis in numerous neurological disorders. The interactions of thiamine with oxidative processes may be part of a spiral of events that lead to neurodegeneration, because reductions in thiamine and thiamine-dependent processes promote neurodegeneration and cause oxidative stress. The reversal of the effects of thiamine deficiency by antioxidants, and amelioration of other forms of oxidative stress by thiamine, suggest that thiamine may act as a site-directed antioxidant. The data indicate that the interactions of thiamine-dependent processes with oxidative stress are critical in neurodegenerative processes.


Inactivation of Escherichia coli phosphoribosylpyrophosphate synthetase by the 2',3'-dialdehyde derivative of ATP. Identification of active site lysines.
The enzyme 5-phosphoribosyl-alpha-1-pyrophosphate (PRPP) synthetase from Escherichia coli was irreversibly inactivated on exposure to the affinity analog 2',3'-dialdehyde ATP (oATP). The reaction displayed complex saturation kinetics with respect to oATP with an apparent KD of approximately 0.8 mM. Reaction with radioactive oATP demonstrated that complete inactivation of the enzyme corresponded to reaction at two or more sites with limiting stoichiometries of approximately 0.7 and 1.3 mol of oATP incorporated/mol of PRPP synthetase subunit. oATP served as a substrate in the presence of ribose-5-phosphate, and the enzyme could be protected against inactivation by ADP or ATP. Isolation of radioactive peptides from the enzyme modified with radioactive oATP, followed by automated Edman sequencing allowed identification of Lys181, Lys193, and Lys230 as probable sites of reaction with the analog. Cysteine 229 may also be labeled by oATP. Of these four residues, Lys193 is completely conserved within the family of PRPP synthetases, and Lys181 is found at a position in the sequence where the cognate amino acid (Asp181) in human isozyme I PRPP synthetase has been previously implicated in the regulation of enzymatic activity. These results imply a functional role for at least two of the identified amino acid residues.


[Localised tetanus with no obvious entry site: about a case in Bamako (Mali)].
Tetanus is a major public health problem in sub-Saharan Africa. Localised tetanus is rare, unlike generalized tetanus which has been sufficiently described in the literature. We report a case of localised tetanus with no obvious entry site managed in the Department of Infectious Diseases in Bamako. The study involved a retired nurse aged 59 years who had not undergone tetanus booster immunisation within the last 10 years, corresponding to the date of her last delivery. She was referred to our Hospital with dysphagia associated with inability to open the buccal cavity. Patient's history was characterized by long-term therapy associated with many specialized consultations without any improvement. The diagnosis of localised tetanus with no obvious entry site was retained after having excluded any other local disorder. Outcome was favorable ten days after adequate management. Underdiagnosed or unknown to health-care providers, localised tetanus may mimic other diseases delaying diagnosis and management. Targeted campaign to build awareness should be implemented in order to improve adherence with immunization schedules.


Maximum distance-gradient for robust image registration.
To make up for the lack of concern on the spatial information in the conventional mutual information based image registration framework, this paper designs a novel spatial feature field, namely the maximum distance-gradient (MDG) vector field, for registration tasks. It encodes both the local edge information and globally defined spatial information related to the intensity difference, the distance, and the direction of a voxel to a MDG source point. A novel similarity measure is proposed as the combination of the multi-dimensional mutual information and an angle measure on the MDG vector field. This measure integrates both the magnitude and orientation information of the MDG vector field into the image registration process. Experimental results on clinical 3D CT and T1-weighted MR image volumes show that, as compared with the conventional mutual information based method and two of its adaptations incorporating spatial information, the proposed method can give longer capture ranges at different image resolutions. This leads to more robust registrations. Around 2000 randomized rigid registration experiments demonstrate that our method consistently gives much higher success rates than the aforementioned three related methods. Moreover, it is shown that the registration accuracy of our method is high.


Immunocytochemical localization of cathepsins B and H in rat liver.
Light and electron microscopic localization of cathepsins B and H in rat liver was investigated by immunoenzyme and protein A-gold techniques. For light microscopy (LM), semi-thin sections of the Epon-embedded material were stained by the immunoenzyme technique after removal of epoxy resin. For electron microscopy (EM), ultra-thin sections of the Lowicryl K4M-embedded material were stained by the protein A-gold technique. By LM, reaction deposits for cathepsins B and H were present in the cytoplasmic granules of parenchymal cells and endothelial cells, and Kupffer cells. The sinus-lining cells and the parenchymal cells showed the similar staining intensity. By EM, gold particles were present exclusively in lysosomes of all the cell types cited above. The same results were obtained from quantitative analysis. In addition, Golgi complexes themselves were mostly negative but some small vesicles on the trans side of them were labeled for these proteinases. The results indicate that cathepsins B and H are present in the lysosomes of rat liver and that these enzymes seem to be transported by small vesicles from endoplasmic reticulum to lysosomes via tubuloreticular network of the trans Golgi region.


Prediction of clinical outcomes from rTMS in depressed patients with lateral visual field stimulation: a replication.
The authors sought to replicate an earlier finding that baseline lateral visual field stimulation, a procedure shown to activate the contralateral hemisphere and induce affective changes, predicted the clinical outcomes of a 10-day course of left-sided rapid transcranial magnetic stimulation (rTMS). For 23 patients there was a significant 1-tailed Pearson correlation between the percent improvement in response to rTMS and their lateralized affective responses to lateral visual field stimulation. These correlations, across the whole group and within genders, were almost identical to those previously reported from 37 patients studied at a different site.


A gene-environment interplay between omega-3 supplementation and APOE ε4 provides insights for Alzheimer's disease precise prevention amongst high-genetic-risk population.
The present study aimed to explore whether and how omega-3 (ω-3) supplementation could interact with genetic factors to modulate cognitive functions, amyloid pathologies, and Alzheimer's disease (AD) risk. A total of 1,670 non-demented participants (mean age 73 years, 47% females, 41% APOE ε4 carriers) were followed up for 10 years. Hierarchical regressions, linear mixed-effects models, and Cox proportional hazards models were used to examine the interaction effects of ω-3 supplementation with APOE ε4 and polygenic hazard scores, after adjusting for age, gender, education, cognitive diagnosis, insomnia, depression, anxiety, and cardiovascular risk score. Individuals who progress to AD during the follow-up tend to take a shorter duration of ω-3 at baseline than those stable, for whom the difference remained significant only amongst APOE ε4 carriers (p < 0.01). The interaction term (APOE ε4 × ω-3) accounted for a significant amount of variance in cognition and cerebral amyloid burden. Long-term ω-3 use protected cognition (especially memory function) and lowered amyloid burden and AD risk only amongst APOE ε4 carriers. Mediation analysis suggested that amyloid pathologies, brain reserve capacities, and brain metabolism mediated the relationships of ω-3 use with memory and global cognition for APOE ε4 (+) carriers. Similar interaction and mediation effects were also indicated amongst high-risk subjects defined by polygenic hazard scores. Long-term ω-3 intake may have a role in AD prevention in genetically at-risk populations.


Cytokines in airway inflammation.
With over 50 potential asthma mediators, cytokines are the latest group of substances which have been investigated for their potential role in this disease. The use of murine models of allergic inflammation has facilitated the investigation of the role of individual cytokines in this response. The use of targeted gene disruption, overexpression of genes and monoclonal antibodies directed against cytokines have allowed a detailed examination of the role cytokines play in IgE production, eosinophil recruitment and bronchial hyperresponsiveness, which are the characteristic features of the asthma phenotype. Despite the introduction of this new methodology, conflicting reports relating to the role of cytokines in allergic inflammation, highlight the complexity of allergic inflammation and challenge the notion that a single cytokine can explain the asthma phenotype.


Applications of CRISPR/Cas gene-editing technology in yeast and fungi.
Genome editing technology has progressed rapidly in recent years. Although traditional gene-editing methods, including homologous recombination, zinc finger endonucleases, and transcription activator-like effector nucleases, have substantial implications for research in genetics and molecular biology, but they have remarkable limitations, including their low efficiency, high error rate, and complex design. A new gene-editing technology, the CRISPR/Cas system, was developed based on studies of archaeal and bacterial immune responses to viruses. Owing to its high target efficiency, simple primer design, and wide applications, the CRISPR/Cas system, whose developers were awarded the Nobel Prize in Chemistry in 2020, has become the dominant genomic editing technology in academia and the pharmaceutical industry. Here, we briefly introduce the CRISPR/Cas system and its main applications for genome engineering, metabolic engineering, and transcriptional regulation in yeast, filamentous fungi, and macrofungi. The polygene and polyploid editing, construction of yeast chromosomes, yeast library creation, regulation of metabolic pathways, and CRISPR activation/CRISPR interference systems are mainly summarized and discussed. The potential applications for the treatment of fungal infections and the further transformation and application of the CRISPR/Cas system in fungi are also proposed and discussed.


Dander is the best epithelial source for dog allergenic extract preparations.
Four different commercially available raw materials of dog epithelia (dander, hair and skin scraping, skin scraping, and whole skin) were compared by means of biochemical methods such as SDS-PAGE and chromatography with the SMART System, immunochemical methods such as RAST and SDS-PAGE immunoblotting, and cutaneous tests. Dander extract clearly exhibited the highest IgE binding and in vivo allergenic activity. The dog major allergens Can fI and the 19-kDa protein were adequately detected only in dander extract. Thus, it is concluded that dander is the most suitable epithelial source for preparing dog allergenic extracts.


High-throughput, high-damage-threshold broadband beam splitter for high-order harmonics in the extreme-ultraviolet region.
We demonstrate a high-throughput and high-damage-threshold beam splitter for high-order harmonics in the soft-x-ray region that uses Si and (or) SiC plates set at Brewster's angle with respect to the pump wavelength. The beam splitters are guaranteed to have a damage threshold of at least 0.8 TW/cm2 (average power density, 0.25 W/cm2) and an attenuation rate of 10(-4)-10(-5) for a 30-fs pump pulse. The measured reflection efficiency at the 27th harmonic (29.6 nm) was 0.56 for Si and 0.45 for SiC. These beam splitters are useful not only for high harmonics but also for longitudinally pumped x-ray lasers.


Safety and efficacy of switching from low molecular weight heparin to dabigatran in patients undergoing elective total hip or knee replacement surgery.
The aim of this study was to assess the safety and efficacy of switching therapy from low molecular weight heparin (LMWH; enoxaparin) to dabigatran for prevention of venous thromboembolic events (VTE) in patients undergoing elective total hip or knee replacement surgery (THR/TKR). This was a prospective, multicenter, open-label, single-arm, observational, study in patients undergoing THR or TKR who were to receive enoxaparin 40 mg for thromboprophylaxis. Enoxaparin was initiated before or after surgery according to local practice, and was switched to dabigatran 220 mg once daily at a time point chosen by the investigator. The coprimary endpoints were major bleeding events, and the composite of symptomatic VTE and all-cause mortality, from last use of enoxaparin to 24 h after last intake of dabigatran. Altogether, 168 (81 THR, 87 TKR) patients were enrolled, of whom 161 received both enoxaparin and dabigatran, 2 received dabigatran only and 5 received enoxaparin only. The median time of the first dabigatran tablet was 24.0 h after the last LMWH dosage and the median number of days on dabigatran treatment was 36 days. No symptomatic VTE or death occurred during the study. One major bleeding event was seen at the surgical site and required treatment cessation. Three minor bleeding events were observed. In the normal clinical setting, switching from LMWH to dabigatran in patients who had undergone THR and TKR was safe and effective in preventing VTE. The reported adverse events and serious adverse events were consistent with the known safety profile for dabigatran. Switching from a subcutaneous to an oral anticoagulant may offer greater convenience in the outpatient setting after discharge. ClinicalTrials.gov identifier NCT01153698.


Nonclinical evaluation of therapeutic cytokines: immunotoxicologic issues.
Cytokines are a diverse family of immunoregulatory proteins whose function has been the focus of intensive research for the past 25 years. Modern molecular biology techniques have resulted in production of these proteins in quantities sufficient to consider them as candidates for drug development. Due to the often pleiotropic nature of their action, in order to design meaningful nonclinical safety assessment studies, the practicing toxicologist must be able to differentiate between an exaggerated, but expected pharmacologic response and toxicity associated with their administration. Species specificity, nontraditional dose-response relationships, antigenicity, and unanticipated immunotoxicity are a few issues that must be considered when designing study protocols and interpreting data. In addition, despite validated methodology to examine the immune system as a potential target organ for toxicity, limitations exist in the ability to predict the autoimmune and allergenic potential of proteins such as these. Despite these challenges and limitations, over the past decade, many of these molecules have found clinical utility to treat a wide variety of diseases.


A study of five year old children born after elective induction of labour.
A controlled retrospective study of some short-term and long-term effects of elective induction of labour was conducted. There did not appear to be any increase in the incidence of maternal complications during labour, or of neonatal problems in the induced-labour group compared with the remainder of the mothers in the hospital. In the follow-up phase of the study, two treatment groups of children who were delivered after amniotomy and amniotomy plus the administration of oxytocin were compared with each other, and with a control group of children born after spontaneous labour. The children were assessed at the age of five years on verbal and non-verbal subtests of a standardized intelligence scale, tests of gross motor and fine motor coordination, and auditory and visual tests. A full physical examination was also performed. No statistically significant differences nor trends of clinical interest were found between the groups on any measure.


Cytotoxic interactions of methylene blue with trypanosomatid-specific disulfide reductases and their dithiol products.
Methylene blue (MB) is known to have trypanocidal activity. We tested the interactions of MB with a number of trypanosomatid-specific molecules of the antioxidant metabolism. At pH 7, trypanothione and other (di)thiols were oxidized to disulfides by the phenothiazine drug. MB inhibited Trypanosoma cruzi trypanothione reductase (TR) (K(i)=1.9 microM), and served as a significant subversive substrate of this enzyme (K(M)=30 microM, k(cat)=4.9s(-1)). With lipoamide dehydrogenase, the second thiol-generating flavoenzyme of T. cruzi, the catalytic efficiency for MB reduction was found to be almost 10(6)M(-1)s(-1). When the system MB-enzyme-molecular oxygen acts as a NAD(P)H-driven redox cycler, a reactive oxygen species, H(2)O(2) or superoxide, is produced in each cycle. Since MB is an affordable, available, and accessible drug it might be tested--alone or in drug combinations--against trypanosomatid-caused diseases of animal and man.


mTORC1 activity is essential for erythropoiesis and B cell lineage commitment.
Mechanistic target of rapamycin (mTOR) is a serine/threonine protein kinase that mediates phosphoinositide-3-kinase (PI3K)/AKT signalling. This pathway is involved in a plethora of cellular functions including protein and lipid synthesis, cell migration, cell proliferation and apoptosis. In this study, we proposed to delineate the role of mTORC1 in haemopoietic lineage commitment using knock out (KO) mouse and cell line models. Mx1-cre and Vav-cre expression systems were used to specifically target Raptorfl/fl (mTORC1), either in all tissues upon poly(I:C) inoculation, or specifically in haemopoietic stem cells, respectively. Assessment of the role of mTORC1 during the early stages of development in Vav-cre+Raptorfl/fl mice, revealed that these mice do not survive post birth due to aberrations in erythropoiesis resulting from an arrest in development at the megakaryocyte-erythrocyte progenitor stage. Furthermore, Raptor-deficient mice exhibited a block in B cell lineage commitment. The essential role of Raptor (mTORC1) in erythrocyte and B lineage commitment was confirmed in adult Mx1-cre+Raptorfl/fl mice upon cre-recombinase induction. These studies were supported by results showing that the expression of key lineage commitment regulators, GATA1, GATA2 and PAX5 were dysregulated in the absence of mTORC1-mediated signals. The regulatory role of mTOR during erythropoiesis was confirmed in vitro by demonstrating a reduction of K562 cell differentiation towards RBCs in the presence of established mTOR inhibitors. While mTORC1 plays a fundamental role in promoting RBC development, we showed that mTORC2 has an opposing role, as Rictor-deficient progenitor cells exhibited an elevation in RBC colony formation ex vivo. Collectively, our data demonstrate a critical role played by mTORC1 in regulating the haemopoietic cell lineage commitment.


Spatial-temporal distribution and pedestrian exposure assessment of size-fractionated particles on crosswalk of urban intersection.
Vehicles tend to produce more pollutants especially particles at an urban intersection than other segments. Meanwhile, pedestrians at an intersection are inevitably exposed to high particle level and suffered from the health problem. Especially, some particles can deposit in different thoracic areas of the respiratory system and cause serious health problems. Hence, in this paper, the particles from 0.3 to 10 μm in 16 channels were measured to compare the spatio-temporal characteristics of them on the crosswalk and the roadside. Based on the roadside of fixed measurements, submicron particles (< 1 μm) are discovered to have a high relation with traffic signal and exhibit a bimodal distribution pattern in the green phase. On the crosswalk of mobile measurements, submicron particles present decreasing trend along the crosswalk while crossing. Additionally, mobile measurements were conducted across six time intervals that correspond to different pedestrian's journey when passing the crosswalk. The results showed that all size particles in the first three journeys present high concentrations than that in other journeys. Furthermore, pedestrian exposure to all 16 channel particles was assessed. The total and regional deposition fractions of these particles in different sizes and age groups are determined. What ought to be paid attention to is that these real-world measurement results contribute to advancing the understanding of pedestrian exposure to size-fractionated particles on crosswalk and assisting the pedestrian to make better informed choice so as to limit particle exposure in these pollution hotspots.


Pain complaint as the first symptom of oral cancer: a descriptive study.
To identify pain as the initial symptom of oral cancer patients. Hospital charts of 1412 patients (1977 to 1998) with oral cancer were reviewed (238 female and 1174 male). Pain was the initial complaint in 19.2% of the sample. Oral cancer (ratio 4:1) and initial pain (ratio 9:1; P= .001) was prevalent in men. There were 12 different complaints of pain: sore throat (37.6%), tongue pain (14.0%), mouth pain (12.9%); pain when swallowing (11.1%), dental pain (5.9%); earache (5.9%); pain in the palate (4.1%); burning mouth (3.3%); gingival pain (2.2%); pain when chewing (1.1%); neck pain (1.1%), and facial pain (0.7%). Pain was associated with advanced TNM staging and location of tumor in the tongue (P= .004) and the tongue/mouth floor (P< .006). There were 12 different descriptions of pain; pain was related to TNM staging in the tongue and the tongue/mouth floor. The data presented reinforce the suggestion that patients with orofacial pain need systematized evaluation and sometimes require an interdisciplinary approach.


Immunohistochemistry of matrix metalloproteinases and their inhibitors in thoracic aortic aneurysms and aortic valves of patients with Marfan's syndrome.
Thoracic aortic aneurysms (TAAs) and valvular insufficiency, the main cardiovascular lesions in Marfan's syndrome, are associated with destruction of connective tissue; however, their pathogenesis remains unclear. To test the hypothesis that changes in the activity of the matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) are responsible for the damage to connective tissue in these lesions, histochemical studies of the immunoreactivity (IR) for MMPs and their tissue TIMPs (MMP-1, MMP-2, MMP-3, MMP-9, TIMP-1, and TIMP-2) were made in TAAs (n = 7) and aortic valves (n = 5) from 7 patients with Marfan's syndrome. All TAAs showed cystic medial necrosis (CMN), with loss of elastic fibers and smooth muscle cells. Extensive areas of myxoid change were found in all aortic valves. Areas of CMN showed no IR for any MMPs or TIMPs. The IR of smooth muscle cells at the borders of areas of CMN was stronger for all MMPs, especially MMP-2 and MMP-9, than in other regions. The surfaces of disrupted elastic fibers showed IR for MMP-2 and MMP-9. Areas of myxoid change showed similar but less pronounced alterations. We hypothesize that the defect in fibrillin-1 in Marfan's syndrome leads to (1) formation of elastin that is abnormally aggregated and more easily degraded by MMPs than is normal elastin, (2) upregulation of the synthesis of MMPs, (3) progressive destruction of connective tissue by these enzymes, and (4) development of TAAs and valvular lesions.


Steady-state kinetic analysis of substrate pair cycling between two enzymes: application to a mediated electron transport between the cytoplasmic membrane and the periplasmic nitrite reductase of Paracoccus denitrificans.
An extended kinetic model is presented for the process catalysed by two enzymes mutually connected by the cycling of two reversibly interconvertible chemically relative species. Expressions are derived for the steady-state velocity, limiting velocity (V) and the half-saturation concentration of the cycling substrate (A(0.5)). It is shown that the velocity depends on the total concentration of cycling substrate hyperbolically if both enzymes have equal activities. Based on these theoretical considerations, an experimental comparison was made between pseudoazurin and cytochrome c(550) as physiological electron transfer mediators for nitrite reduction in an in vitro reconstituted part of the respiratory chain of Paracoccus denitrificans. Pseudoazurin exhibited 1.7-fold higher V and 14-fold higher A(0.5) than cytochrome c(550) under the experimental conditions used (20 mM Tris chloride, pH 7.3, 30 degrees C).


Functional gland-preserving surgery for submandibular gland pleomorphic adenoma.
Conventional surgery for submandibular gland tumour is total excision of the gland affected by the tumour. Gland-preserving surgery is commonly used for benign tumours arising in the parotid gland but not in the submandibular gland. This study evaluated long-term oncological and functional outcomes after gland-preserving surgery in patients with pleomorphic adenoma of the submandibular gland via the submental approach. This longitudinal study included 105 consecutive patients with submandibular gland pleomorphic adenoma who underwent the gland-preserving surgery combined with the en-bloc resection of tumours via the submental approach. Salivary scintigraphy was performed 6 months after surgery, and ultrasonography was regularly followed. Intraoperative findings, postoperative complications, cosmetic and salivary functions, and tumour recurrence were assessed in these patients. Median tumour size and submental incision length were 2.0 and 3.3 cm, respectively. Median operation time and amount of blood loss were 25 min and 18.5 mL, respectively. None had marginal or lingual nerve paralysis and most patients were satisfied with incision scar and facial contour. The salivary function of the affected gland was equal to that of the unaffected gland. One patient (0.9%) had single nodular recurrence 54 months after surgery and others had no recurrence for follow-up of median 96 months. Pleomorphic adenoma of the submandibular gland can be safely removed by the gland-preserving surgery via the submental approach which has operation time, cosmetic and functional benefits with compromising oncological outcomes.


Loss of hepatic manganese transporter ZIP8 disrupts serum transferrin glycosylation and the glutamate-glutamine cycle.
ZIP8, encoded by SLC39A8, is a membrane transporter that facilitates the cellular uptake of divalent biometals including zinc (Zn), manganese (Mn), and iron (Fe). The hepatic system has long been accepted as the central modulator for whole-body biometal distribution. Earlier investigations suggest the propensity of ZIP8 to prioritize Mn influx, as opposed to Fe or Zn, in hepatocytes. Hepatic ZIP8 Mn transport is crucial for maintaining homeostasis of various Mn-dependent metalloenzymes and their associated pathways. Herein, we hypothesize that a drastic decrease in systemic Mn, via the loss of hepatic ZIP8, disrupts two unique cellular pathways, post-translational glycosylation and the glutamate-glutamine cycle. ZIP8 liver-specific knockout (LSKO) mice were chosen in an attempt to substantially decrease whole-body Mn levels. To further elucidate the role of Mn in serum glycosylation, a Mn-deficient diet was adopted in conjunction with the LSKO mice to model a near-complete loss of systemic Mn. After the treatment course, transferrin sialylation profiles were determined using imaged capillary isoelectric focusing (icIEF). We also investigated the role of Mn in the glutamate-glutamine cycle; the conversion of glutamate to glutamine in F/F and LSKO mice was assessed by the glutamine/glutamate ratio in cerebrospinal fluid (CSF) via HPLC-MS. An open-field study was ultimately conducted to check if these mice displayed atypical behavior. Two major biological pathways were found to be significantly altered due to the loss of hepatic ZIP8. We identified a disparity between F/F and LSKO transferrin sialylation profiles that were exacerbated under a Mn-deficient diet. Additionally, we discovered a neurotransmitter imbalance between the levels of glutamine and glutamate, exclusive to LSKO mice. This was characterized by the decreased glutamine/glutamate ratio in CSF. Secondary to the neurotransmitter alteration, LSKO mice exhibited an increase in locomotor activity in an open-field. Our model successfully established a connection between the loss of hepatic ZIP8 and two Mn-dependent cellular pathways, namely, protein glycosylation and the glutamate-glutamine cycle.


Functionally distinct subpopulations of cord blood CD34+ cells are transduced by adenoviral vectors with serotype 5 or 35 tropism.
Adenovirus serotype 5 (Ad5)-based vectors can be retargeted with fiber receptor specificity of serotype 35 adenovirus (Ad5F35) and thereby bypass the paucity of the coxsackie and adenovirus receptor (CAR) on hematopoietic cells by utilizing CD46 as cellular receptor. The gene transfer efficiency into NOD/SCID repopulating cells by an Ad5F35-GFP vector was investigated in comparison with its corresponding Ad5-GFP vector. Cord blood CD34+ cells were transduced following overnight culture under serum-free conditions supported by early acting cytokines. In agreement with previous findings, the Ad5F35-GFP vector showed significant superiority to the Ad5-GFP vector in gene transfer into cells with primitive immunophenotype. However, the Ad5F35-GFP vector allowed efficient gene transfer into both dividing and nondividing CD34+ cells, whereas the Ad5-GFP vector preferentially allowed gene transfer into dividing cells expressing lower levels of CD34 antigen, which correlated with high levels of CAR expression. The sorted GFP+ cells following Ad5F35-GFP transduction at relatively low multiplicity of infection consistently reconstituted the NOD/SCID mouse bone marrow with multilineage differentiation. In contrast, the GFP+ cells following Ad5-GFP transduction were nearly devoid of reconstitution capacity. Thus, Ad5F35 vectors encoding functional genes can facilitate transient genetic manipulation of human NOD/SCID repopulating cells.


Sufentanil versus fentanyl: efficacy and patient satisfaction with intrathecal pain management.
This study compared fentanyl vs. sufentanil in intrathecal pain pumps. H1: both reduce patient subjective pain ratings. H2: sufentanil is more effective than fentanyl. H3: overall satisfaction with pain control is greater with sufentanil. This is an archival study of patients in tertiary pain management (N = 97, mean age = 58.77, standard deviation = 14.88). Pain was measured using the subjective units of discomfort scale. Satisfaction with pain control/relief was measured by asking patients each visit if they are satisfied with pain management and is recorded in a "yes"/ "no" manner. Pain ratings were analyzed with repeated measures analysis of variance and satisfaction was analyzed with chi square. Sufentanil was found to be marginally more effective, but both medications controlled a significant degree of variance in pain reduction over time. A significantly greater number of patients maintained on sufentanil were satisfied with care than patients on fentanyl.


Photopatterned Membranes and Chemical Gradients Enable Scalable Phenotypic Organization of Primary Human Colon Epithelial Models.
Biochemical gradients across the intestinal epithelium play a major role in governing intestinal stem cell compartmentalization, differentiation dynamics, and organ-level self-renewal. However, scalable platforms that recapitulate the architecture and gradients present in vivo are absent. We present a platform in which individually addressable arrays of chemical gradients along the intestinal crypt long axis can be generated, enabling scalable culture of primary in vitro colonic epithelial replicas. The platform utilizes standardized well plate spacing, maintains access to basal and luminal compartments, and relies on a photopatterned porous membrane to act as diffusion windows while supporting the in vitro crypts. Simultaneous fabrication of 3875 crypts over a single membrane was developed. Growth factor gradients were modeled and then experimentally optimized to promote long-term health and self-renewal of the crypts which were assayed in situ by confocal fluorescence microscopy. The cultured in vitro crypt arrays successfully recapitulated the architecture and luminal-to-basal phenotypic polarity observed in vivo. Furthermore, known signaling regulators (e.g., butyrate and DAPT) produced measurable and predictable effects on the organized cell compartments, each decreasing crypt proliferation in the basal regions to negligible values. This platform is readily adaptable to the screening of tissue from individual patients to assay the impact of food and bacterial metabolites and/or drugs on colonic crypt dynamics. Importantly, the cassette is compatible with a wide range of sensing/detection modalities, and the developed fabrication methods should find applications for other cell and tissue types.


Supersensitization of neurochemical responses by L-DOPA and dopamine receptor agonists in the striatum of experimental Parkinson's disease model rats.
We studied the mechanisms of dopamine receptor agonist- and L-DOPA-mediated supersensitization in experimental Parkinson's disease model rats, by measuring in vivo acetylcholine (ACh) release, GTPase activities, and mRNA expression in the striatum of 6-hydroxydopamine-treated rats. D1 agonist (SKF38393) and D2/D3 agonists (bromocriptine and quinpirole) showed more potent stimulation or inhibitions on ACh release in the model rat than in the control. However, quinpirole-evoked stimulation of GTPase activity was enhance in the model rats, compared to the control, while there was no significant enhancement of the bromocriptine-evoked stimulation. On the other hand, L-DOPA at 0.3-10 pM showed a biphasic action including significant inhibition on the GTPase activity in the lesioned striatal membranes, but not in the control. In the RNAase protection assay, neither D1, D2, Gi1 alpha, GoA alpha nor Gs alpha mRNA expression in the model was significantly different from the control. These findings suggest that there is supersensitization of D1 and D2/D3 receptors in the experimental Parkinson's disease model, while the upregulation of their receptors or GTP-binding proteins (G-proteins) to be coupled to their receptors is unlikely involved in major parts of such mechanisms. In addition, the present report provides the first evidence that L-DOPA mediates neurochemical responses in the plasma membranes, possibly through its receptor.


Identification of LOX as a candidate prognostic biomarker in Glioblastoma multiforme.
Glioblastoma multiforme (GBM) is the most malignant type of glioma. GBM tumors grow rapidly, have a high degree of malignancy, and are characterized by a fast disease progression. Unfortunately, there is a lack of effective treatments. An effective strategy for the treatment of GBM would be to identify key biomarkers correlating with the occurrence and progression of GBM and developing these biomarkers into therapeutic targets. In this study, using integrated bioinformatics analysis, we identified differentially expressed genes (DEGs), including 130 genes that were upregulated in GBM compared to normal brain tissue, and 128 genes that were downregulated in GBM. Based on Gene Ontology enrichment analysis and Kyoto Encyclopedia of Genes and Genomes pathway analysis, these genes were associated with regulation of tumor cell adhesion, differentiation, morphology in GBM and were mainly enriched in Complement and coagulation cascades pathway. The Search Tool for the Retrieval of Interacting Genes (STRING) database was used to construct a Protein-Protein Interaction network. Ten hub genes were identified, including FN1, CD44, MYC, CDK1, SERPINE1, COL3A1, COL1A2, LOX, POSTN and EZH2, all of which were significantly upregulated in GBM, these results were confirmed by oncomine database exploration. Alteration analysis of hub genes found that patients with alteration in at least one of the hub genes showed shorter median survival times (p = 0.013) and shorter median disease-free survival times (p = 2.488E-3) than patients without alterations in any of the hub genes. Multiple tests for survival analysis showed that among individual hub genes only expression of LOX was correlated with patient survival (P < 0.05).GDS4467 data set was used to analyze the expression of LOX in gliomas with different degrees of malignancy, and it was found that the expression level of LOX was positively correlated with the malignant degree of gliomas.By analyzing GDS 4535 data set showed that the expression level of LOX was positively correlated with the differentiation degree of GBM cells CONCLUSION: This research suggests that FN1, CD44, MYC, CDK1, SERPINE1, COL3A1, COL1A2, LOX, POSTN and EZH2 are key genes in GBM. However, only LOX is correlated with patient survival and promotes glioblastoma cell differentiation and tumor recurrence. LOX may be a candidate prognostic biomarker and potential therapeutic target for GBM.


Nationwide survey on activities of regional core centers for the management of liver disease in Japan: Cumulative analyses by the Hepatitis Information Center 2009-2017.
Regional core centers for the management of liver disease, which are located in every prefecture in Japan, not only take the lead in hepatitis care in their respective regions, but also serve a wide range of other functions, such as education, promotion of hepatitis testing, treatment, and research. Since fiscal year 2010, the Hepatitis Information Center has conducted surveys of regional core centers throughout Japan regarding information about their facilities, programs for patient support, training, and education of medical personnel. By compiling and analyzing the results of these surveys, we have elucidated the status of regional core centers and the issues they currently have. We found that regional core centers have come to play widely varied roles in hepatitis treatment and have expanded their programs. These surveys also suggest that uniform accessibility of hepatitis treatment has been implemented throughout Japan. To continue serving their diverse roles, regional core centers require further development of hepatitis care networks that include specialized institutions, primary care physicians, and local and central governments; as well as collaboration with other professions and groups.


An active site mutant of Escherichia coli cyclopropane fatty acid synthase forms new non-natural fatty acids providing insights on the mechanism of the enzymatic reaction.
We have produced and purified an active site mutant of the Escherichia coli cyclopropane fatty acid synthase (CFAS) by replacing the strictly conserved G236 within cyclopropane synthases, by a glutamate residue, which corresponds to E146 of the homologous mycolic acid methyltransferase, Hma, producing hydroxymethyl mycolic acids. The G236E CFAS mutant had less than 1% of the in vitro activity of the wild type enzyme. We expressed the G236E CFAS mutant in an E. coli (DE3) strain in which the chromosomal cfa gene had been deleted. After extraction of phospholipids and conversion into the corresponding fatty acid methyl esters (FAMEs), we observed the formation of cyclopropanated FAMEs suggesting that the mutant retained some of the normal activity in vivo. However, we also observed the formation of new C17 methyl-branched unsaturated FAMEs whose structures were determined using GC/MS and NMR analyses. The double bond was located at different positions 8, 9 or 10, and the methyl group at position 10 or 9. Thus, this new FAMEs are likely arising from a 16:1 acyl chain of a phospholipid that had been transformed by the G236E CFAS mutant in vivo. The reaction catalyzed by this G236E CFAS mutant thus starts by the methylation of the unsaturated acyl chain at position 10 or 9 yielding a carbocation at position 9 or 10 respectively. It follows then two competing steps, a normal cyclopropanation or hydride shift/elimination events giving different combinations of alkenes. This study not only provides further evidence that cyclopropane synthases (CSs) form a carbocationic intermediate but also opens the way to CSs engineering for the synthesis of non-natural fatty acids.


[Idiopathic nephrotic syndrome and the HLA allele. Prevalence of age-related DR7].
High prevalence of HLA DR7 was well documented in idiopathic nephrotic syndrome (INS) of childhood. Our data in 43 INS, child and adult patients, did not show any significant difference from control group, specially for HLA DR7. Moreover a significant difference occurred when the data was compared in child and adult INS patients. HLA DR7 was more frequent when the nephrotic syndrome appeared before 15 years old. There was no correlation between the presence of HLA DR7 and those of allergy history or increased serum IgE level, on the opposite with previous data. The presence of DR7 would favor its onset at an earlier age. Alternatively, the difference in DR7 prevalence could be an additional argument for considering that the corticosensitivity and negative histological criteria might collect, under the same name, different glomerulopathies.


Treatment with an ascochlorin derivative, AS-6 increases 45Ca2+ binding on the plasma membrane of adipocytes in db/db mice.
Genetically obese diabetic mice (db/db) have greatly diminished 45Ca2+ binding on the plasma membranes of the adipocytes (45-55%) compared with their lean littermates. Treatment for 1 week with a diet admixture of AS-6 (0.1% in the diet) significantly restored the binding to a level comparable to the lean littermates. The addition of AS-6 in vitro had no effect on the binding, which eliminates the possibility that AS-6 is a Ca2+ ionophore. The results suggest that AS-6 treatment enhances the Ca2+ binding by causing structural alteration(s) in the membranes.


Radioimmunotherapy--a potential novel tool for pancreatic cancer therapy?
Pancreatic cancer is one of the most severe cancers and is predicted to rise up to the number two cancer killer by 2030. The ineffective treatment options available and that the cancer is silent until very late in its course are the main reasons for the poor outcome of the disease. Surgery is the only curative option but only available for 10-15 % of the patients, but even then many relapse due to metastases. Many new treatments are under way, and one of the promising ones is radioimmunotherapy (RIT). This review includes clinical trials with RIT in pancreatic cancer as well as a review of adverse events observed during treatment of other solid tumors. Additionally, preclinical studies are reviewed with emphasis on effect, adverse events, the tumor targeting as well as isotope function. Four clinical trials with pancreatic cancer have been conducted with positive results, and one phase III trial is underway. The use of RIT in patients with solid tumors has proven to be well tolerated, and the adverse effects are almost exclusively hematological. Multiple targets and isotopes have been evaluated preclinically, alone, or in combination with existing drug options. Smaller tumors have in several studies completely regressed, while larger ones have stabilized or progressed more slowly. Pancreatic cancer is one of the solid tumors where RIT have reached the longest. The tumor heterogeneity will most likely leave room for more than one treatment option, and several aspiring therapies are under way. RIT may become part of multimodality tumor-directed therapy for pancreatic cancer.


Epoxyeicosatrienoic acid prevents maladaptive remodeling in pressure overload by targeting calcineurin/NFAT and Smad-7.
Emerging evidence demonstrates that epoxyeicosatrienoic acids (EETs) as important active eicosanoids that regulate cardiovascular homeostasis, but the mechanisms underlying its favorable anti-hypertrophic benefits in overpressure model remain obscure. Four weeks after transverse aortic constriction (TAC), TAC mice developed maladaptive cardiac hypertrophy and consequent cardiac failure. Conversely, a cardiotropic adeno-associated viral vector (AAV9) encoding CYP2J2 prevented transverse aortic constriction-induced cardiac hypertrophy with preserved ejection fraction. EET also conferred protection against phenylephrine-induced hypertrophy in H9c2 cardiomyoblasts. Further investigations indicate CYP2J2/EET exerts protection against cardiac hypertrophy through opposing the increase of intracellular Ca2+ level and Ca2+-mediated calcineurin/NFATc3 signaling. Meanwhile, extended myocardial fibrosis in TAC mice was also effectively abolished with the administration of AAV9-2J2. Intriguingly, TAC mice display activated TGF-β/Samd-3 signaling with decreased Smad-7 expression, whereas AAV9-2J2 attenuated the phosphorylation of Smad-3 without altering TGF-β expression, whilst preservation of Smad-7. Subsequently, the differentiation of cardiac fibroblasts into myofibroblasts in the presence of TGF-β1 stimulation was significantly disrupted with EET treatment, accompanied by declined Smad-3 activation and collagen production, whereas inhibition of Smad-7 with SiRNA Smad-7 substantially abrogated these effects of EET on cardiac fibroblasts. EET has synergistic actions on cardiomyocytes and cardiac fibroblasts, preventing cardiac hypertrophy through inhibition of Ca2+-mediated calcineurin/NFATc3 signaling cascades, and ameliorating myocardial fibrosis dependent on Smad-7. This work further extends the potential mechanisms of EET, providing a novel therapeutic approach for the treatment of pathological remodeling and heart failure.


Effects of high and fluctuating pressure on microbial abundance and activity in a water hydraulic system.
The effects of high and fluctuating pressure up to 220 bar on microbial growth and activity were determined in a pilot-scale water hydraulic system. An increase in the pipeline pressure from 70 to 220 bar decreased the total and the viable cell number in the pressure medium from 2.2(+/-0.5)x10(5) to 4.9(+/-1.5)x10(4) cells/ml and from 5.7(+/-2.8)x10(4) to 1.3(+/-0.7)x10(4) cfu/ml, respectively. Microbial attachment in the non-pressurised tank of the hydraulic system increased with increasing pipeline pressure [from 1.0(+/-0.3) to 3.8(+/-2.7)x10(5) cells/cm(2) on stainless steel]. The phosphatase, aminopeptidase and beta-glucosidase activities in the pressurised medium were between 0.02 and 1.4 micromol/lh ( V(max)) and decreased in response to increasing pipeline pressure. The alpha-glucosidase activity was detected only at 70 bar and the glucuronidase activity only occasionally. Based on principal component and cluster analyses, both the pressure applied and the original filling water quality affected substrate utilisation patterns. This study demonstrated the capability of freshwater bacteria to tolerate high and fluctuating pressure in a technical water system. Microbial survival was due to attachment and growth on the surfaces of the non-pressurised components and the nutrient flux released by cell lysis in the pressurised components. In summary, high pressures in water hydraulic systems do not prevent potential microbiologically related operational problems.


Effects of racket moment of inertia on racket head speed, impact location and shuttlecock speed during the badminton smash.
How the racket properties impact performance of the badminton smash is relatively unknown, and further insight could help players/coaches select the most appropriate racket. Three-dimensional position data of the racket and shuttlecock were collected (500 Hz) for 20 experienced badminton players performing a series of forehand smashes with five swingweight ([Formula: see text]) perturbed rackets, ranging from 85-106 kg·cm2. [Formula: see text] was calculated using a balance board and simple pendulum method, and modal analysis was performed using laser vibrometry to capture the fundamental frequency and distal node location for each racket. As [Formula: see text] increased a reduction in racket head speed was found with on average a 0.7 m·s-1 decrease per 5 kg·cm2 increase in [Formula: see text], however this did not lead to slower shuttlecock speeds. The impact location tended to move closer to the tip as the fundamental frequency node moved closer to the tip (as [Formula: see text] increased), providing some evidence that participants may subconsciously strike the shuttlecock at the node location to provide desirable sensory feedback. The increase in racket head speed but not shuttlecock speed was likely due to the distal increase in longitudinal impact location as [Formula: see text] increased, as well as an increase in effective mass for a given impact location. Additionally, removal of the deformation component (additional racket head speed due to the racket noticeably bending and recovering) of racket head speed increased the effect size of the relationship with [Formula: see text], where rackets with greater [Formula: see text] had larger deformation velocities. The research provides further insight into the smash performance characteristics of experienced badminton players, particularly based on racket properties. Further research is required to confirm the coincidence between node location and longitudinal impact location.


A roundtable on responsible innovation with autologous stem cells in Australia, Japan and Singapore.
We report on a roundtable event hosted in Singapore that sought to identify some of the ethical and regulatory challenges in translating autologous cell-based interventions, particularly those claiming to involve stem cells, into safe and effective therapies and to propose some solutions to encourage responsible innovation with these products. Challenges are identified in the three areas of cell manufacturing and processing, innovative uses of autologous cells in clinical practice and standards of evidence. Proposed solutions are discussed within a co-operative model of statutory laws and regulations that can enable product development with autologous cells and professional codes and standards that can encourage ethical conduct in clinical practice. Future research should be directed toward establishing regional networks for the development of internationally consistent standards in manufacturing and ethical codes of conduct for innovating with stem cells, and other autologous cells, and fostering ongoing exchange between jurisdictions.


Altered glutamate and GABA release within thalamocortical circuitry in metabotropic glutamate receptor 4 knockout mice.
The metabotropic glutamate receptor 4 is highly expressed presynaptically on thalamocortical neurons that are involved in the pathogenesis of generalized absence seizures. Mutant mice devoid of metabotropic glutamate receptor 4 are completely resistant to absence seizures induced by low doses of GABA type A receptor antagonists. The purpose of this study was to test the hypothesis that there is altered glutamate and GABA release within thalamocortical circuitry in mice devoid of metabotropic glutamate receptor 4. Extracellular GABA and glutamate release were determined in ventrobasal thalamus, the nucleus reticularis thalami and laminae I-III, and IV-VI of cerebral cortex (laminae I-III of cerebral cortex, and laminae IV-VI of cerebral cortex) using in vivo microdialysis techniques on awake, free moving mice. A significant increase of both basal and K(+)-evoked glutamate release was detected in the ventrobasal thalamus, the nucleus reticularis thalami and laminae IV-VI of cerebral cortex of mice devoid of metabotropic glutamate receptor 4 mice. There also was a significant increase in both basal and K(+)-evoked GABA release in the mice devoid of metabotropic glutamate receptor 4, but a significant decrease of GABA release in laminae IV-VI of cerebral cortex. However, there was no alteration of either GABA or glutamate release in laminae I-III of cerebral cortex, cortical laminae that are not involved in absence seizures. These data indicate that deletion of the metabotropic glutamate receptor 4 gene results in a selective perturbation of glutamate and GABA release within the thalamocortical circuitry involved in the pathogenesis of absence seizures.


Holothuria leucospilota Polysaccharides Improve Immunity and the Gut Microbiota in Cyclophosphamide-Treated Immunosuppressed Mice.
Immunosuppression is one of the major risk factors for a series of diseases, such as tumor, rheumatoid arthritis, and microbial infection. Various natural products have attracted wide attention due to their immunomodulatory activities. Herein, the study investigates the regulation of Holothuria leucospilota polysaccharides (HLP) in immunosuppressed mice. Eight-week-old female BALB/c mice are injected intraperitoneally with cyclophosphamide (80 mg kg-1 body weight day-1 ) to establish the immunosuppressive model. After 12 days of HLP treatment, the immune organ indexes, serum cytokines, and immunoglobulin levels are significantly increased in immunosuppressed mice (p < 0.05). Real-time fluorescent quantitative polymerase chain reaction and western blotting analysis find that HLP improves the immune factors, T-cell markers, and Toll-like receptors (TLR) pathway-related proteins expression. Simultaneously, HLP significantly increases the short-chain fatty acids concentration and regulates the gut microbiota composition (p < 0.05). Furthermore, the metagenomics analysis shows that HLP increases the levels of functional genes involved in amino acid metabolism, carbohydrate metabolism, and growth activity of the gut microbiota. HLP intervention improves the mice's immune function, and the beneficial effects are closely associated with intestinal homeostasis regulation and TLR pathway activation. This study suggests the potential for HLP as prebiotics in novel immunopotentiators development.


Acute toxicity profile in prostate cancer with conventional and hypofractionated treatment.
To compare the acute toxicities in radical treatment of prostate cancer between conventional schedule (C-ARM) with 78 Gy/39 fractions and hypofractionation conformal treatment (H-ARM) with 69 Gy/23 fractions. This prospective double arm study consisted of 217 patients with prostate cancer, 112 in H-ARM and 105 in C-ARM arm. C-ARM received conventional six- field conformal radiotherapy with 78 Gy in 39 fractions while H-ARM received hypofractionation with 69 Gy in 23 fractions. Weekly assessment of acute reactions was done during treatment and with one, and 3 months using RTOG scale. Univariated analysis was performed to evaluate differences between the incidences of acute reaction in the treatment arms. Variables with p value less than 0.1 were included in the multivariated logistic regression. There was no difference between H-ARM versus C-ARM for severity and incidence in genitourinary (GU) and gastrointestinal (GI) acute toxicity. During the treatment comparing H-ARM with C-ARM no differences was observed for GI toxicity (grade 0-3; H-ARM=45.5%, 34%, 18.7% and 1.8% versus C-ARM=47.6%, 35.2%, 17.2% and 0). For acute GU toxicity no difference was detected between H-ARM (grade 0-3; 22.3%, 54.5%, 18.7% and 4.5%) and C-ARM (grade 0-3; 25.8%, 53.3%, 17.1% and 3.8%).At the 3- months follow-up, persistent Grade> =2 acute GU and GI toxicity were 2.5% and 1.8% in H-ARM versus 5.7% and 3% in C-ARM (p>0.05). In univariated and multivariated analyses, there was not any dosimetric predictor for GI and GU toxicity. Our data demonstrate that hypofractionated radiotherapy achieving high biological effective dose using conformal radiotherapy is feasible for prostate cancer, being well tolerated with minimal severe acute toxicity.


The endocrine background of human renal cell carcinoma. II. Attempt to induce the R-5020-binding components by oestrogens.
In an attempt to elevate the concentration of progestin-binding components in human renal cell carcinoma, 24 patients were treated prior to tumour nephrectomy with various doses of oestrogens, potent inducers of progestin receptor synthesis. Low-dose oestrogen treatment was found to be insufficient to significantly induce progestin-binder synthesis in the tumour tissue. In the carcinoma of patients receiving high-dose oestrogen treatment, the average concentration of R-5020-binding sites did not differ from that of the low-dose group. However, 2 out of these patients (12.5%) exhibited R-5020-binder concentrations in the tumour tissue which exceeded the highest quantities of progestin-binding components found in 88 untreated controls previously analysed in our laboratory.


Effects of intratracheal administration of bleomycin on GSH-shuttle enzymes, catalase, lipid peroxidation, and collagen content in the lungs of hamsters.
Intratracheal administration of one unit of bleomycin increased significantly the glucose-6-phosphate dehydrogenase activity to 147, 135, 163, 207, 278, and 148% of the control at 1, 2, 4, 7, 14, and 21 days post-treatment, respectively. The glutathione peroxidase activity was significantly increased to 147 and 124% of the control at 7 and 14 days post-treatment, respectively. The glutathione reductase and catalase activities were significantly elevated to 132, 174, and 145% of the control in the former and 160, 213, and 160% of the control in the latter case at 7, 14, and 21 days post-treatment, respectively. The amount of thiobarbituric acid reacting products as an index of lipid peroxidation was found to be significantly increased to 251, 248, and 253% of the control at 4, 7, and 14 days after treatment, respectively. The amount of total lung collagen in treated hamsters was significantly increased to 140, 197, and 245% of the control at 7, 14, and 21 days post-treatment, respectively. These findings indicate that the increases in the activities of antioxidant enzymes in lungs of bleomycin-treated hamsters result from the multiple cyto-dynamic events such as infiltration of inflammatory cells, and reparative and proliferative processes which generally follow lung injury caused by any oxidant.


Coding assignments of the genome of adult diarrhea rotavirus.
Adult diarrhea rotavirus (ADRV) has caused epidemics of diarrhea in China since 1982 and remains the only group B rotavirus associated with widespread disease in humans. We recently characterized the proteins of ADRV and have now proceeded to identify the gene segments encoding each protein. Viral RNA transcripts were synthesized in vitro with the endogenous viral RNA polymerase and separated by electrophoresis in agarose. The individual transcripts were translated in a cell-free system using nuclease-treated rabbit reticulocyte lysates. The translation products were compared with polypeptides found in purified virus and were characterized by SDS-PAGE, immunoprecipitation, and Western blot analysis using antisera to double- and single-shelled virions, virus cores, and monoclonal antibodies. Furthermore, individual RNA transcripts were hybridized to total dsRNA to determine their genomic origin. Based on this analysis, the core polypeptides VP1, VP2 and VP3 are encoded by segments 1, 2, and 3, respectively. The main polypeptides in the inner capsid, VP6, and the outer capsid, VP4 and VP7, are encoded by segments 6, 4, and 8 respectively. Segments 5, 7, and 9 code for 60, 45, and 30 kDa nonstructural polypeptides. Two other nonstructural polypeptides (24 and 25 kDa) are derived from gene segment 11. Gene segment 10 codes for a 26 kDa polypeptide that is precipitated with serum to ADRV and may be a structural protein VP9. With this exception, gene coding assignments of ADRV are comparable to those of the group A rotaviruses. Our results have clear implications for further work in cloning, sequencing, and expression genes of ADRV and can provide direction towards understanding the origin and the evolution of this virus.


Some comments on mapping from disease-specific to generic health-related quality-of-life scales.
An article by Lu et al. in this issue of Value in Health addresses the mapping of treatment or group differences in disease-specific measures (DSMs) of health-related quality of life onto differences in generic health-related quality-of-life scores, with special emphasis on how the mapping is affected by the reliability of the DSM. In the proposed mapping, a factor analytic model defines a conversion factor between the scores as the ratio of factor loadings. Hence, the mapping applies to convert true underlying scales and has desirable properties facilitating the alignment of instruments and understanding their relationship in a coherent manner. It is important to note, however, that when DSM means or differences in mean DSMs are estimated, their mapping is still of a measurement error-prone predictor, and the correct conversion coefficient is the true mapping multiplied by the reliability of the DSM in the relevant sample. In addition, the proposed strategy for estimating the factor analytic mapping in practice requires assumptions that may not hold. We discuss these assumptions and how they may be the reason we obtain disparate estimates of the mapping factor in an application of the proposed methods to groups of patients.


Small platform stress attenuates the anxiogenic effect of diazepam withdrawal in the plus-maze test.
The effect of stress on the behavioural changes caused by diazepam withdrawal was studied in mice. Diazepam (2.5 mg/kg) or vehicle was injected intraperitoneally twice a day for 2 weeks. When the last vehicle or diazepam injection had been administered to the mice, 12 h later, the mice were subjected to small platform stress exposure or left in their home cages. Small platform stress was induced by placing mice on small platforms (3.5 cm diameter) surrounded by water for 24 h. This experimental model contains several factors of stress like rapid eye movement (REM) sleep deprivation, isolation, immobilization and falling into the water. The plus-maze test was carried out with stressed mice as well as with mice not subjected to stress. Small platform stress induced an anxiolytic effect and diazepam withdrawal--an anxiogenic effect in the plus-maze test. Small platform stress also attenuated the anxiogenic effect of diazepam withdrawal. On the basis of this data it was proposed that small platform stress counteracts the anxiogenic effect of diazepam withdrawal in the plus-maze test. It was also proposed that the effect of stress on the signs of benzodiazepine withdrawal depends on the characteristics and duration of stress.


Identification of a new small molecule chemotype of Melanin Concentrating Hormone Receptor-1 antagonists using pharmacophore-based virtual screening.
MCH receptor is a G protein-coupled receptor with two subtypes R1 and R2. Many studies have demonstrated the role of MCH-R1 in feeding and energy homeostasis. It has been proven that oral administration of small molecule MCH-R1 antagonists significantly reduces food intake and causes a dose-dependent weight loss. In this study, two ligand-based pharmacophores were developed and validated based on recently published MCH-R1 antagonists with diverse structures. Successful pharmacophores had one hydrogen bond acceptor, one positive ionizable, one ring aromatic and two or three hydrophobic groups. These 3D-QSAR models were used for virtual screening of the ZINC chemical database resulting in the identification of nine compounds with more than 50% displacement of radiolabeled MCH at a 20 μM concentration. Moreover, four of these compounds showed antagonistic activities in Aequorin functional assay, including MH-3 which is the first MCH-R1 antagonist based on a diazaspiro[4.5]decane scaffold. The most active compounds were also docked into our previously published MCH-R1 homology model to gain insights into their binding determinants. These compounds could represent a viable starting scaffold for the design of potent MCH-R1 antagonists with improved pharmacokinetic properties as an effective treatment for obesity.


Infertility and pregnancy after breast cancer: current knowledge and future perspectives.
Fertility impairment induced by adjuvant treatments and potential risk associated with pregnancy, are major concerns of young pre-menopausal patients with early breast cancer. Although current evidences suggest that pregnancy does not negatively affect prognosis, a low rate (3-8%) of pregnancy after breast cancer has been reported. Among the potential causes of such a low rate there are a high chance of spontaneous abortions (25%) as well as the fertility impairment induced by adjuvant treatments. No standard strategy to preserve fertility in breast cancer patients is available so far. Experimental approaches include cryopreservation strategies, and use of gonadotropin-releasing hormone (GnRH) agonists to render germinal epithelium quiescent and less sensitive to the chemotherapy cytotoxicity. Here, we reviewed current knowledge about incidence and risks of pregnancy after breast cancer, risks of ovarian failure after adjuvant treatments and experimental strategies aiming to preserve ovarian function and fertility in young breast cancer patients.


Isolation and characterization of further defective clones of a temperature sensitive mutant (ts-1) of respiratory syncytial virus.
After exposure of the temperature sensitive ts-1 mutant of respiratory syncytial virus to the chemical mutagne, nitrosoguanidine (NG), 2 clones of virus were recovered which were more temperature sensitive and stable genetically than the ts-1 mutant. The initial criterion used for selection of the 2 clones was decreased ability to produce plaques at 36 degrees C. Subsequently it was shown that the 2 clones grew less well at the restrictive temperatures of 37 degrees and 38 degrees C than did the ts-1 parent. Peak titers of the NG derived clones were decreased 10--30 fold at 37 degrees C and over 100-fold at 38 degrees C compared to ts-1. Complementation analysis indicated that the NG mutants retained the same complementation pattern as the ts-1 parent.


Environmental constraints on the invasion of Triadica sebifera in the eastern United States: an experimental field assessment.
Identifying the environmental constraints that affect the distribution of an invasive species is fundamental to its effective control. Triadica sebifera (Chinese tallow tree) has invaded the southeastern United States, but its potential for further range and habitat extension has been unresolved. We explored experimentally environmental factors in macro- and microhabitats that affect its persistence at five widely separated sites along the Atlantic seaboard of the United States and at two sites inland; three sites occur well beyond the tree's current range. At each site, seeds and young vegetative plants (0.5-0.65 m tall) of T. sebifera were placed in four microhabitats (closed-canopy upland, closed-canopy lowland, open-canopy upland, and open-canopy lowland). Plant growth, leaf CO(2) assimilation rates, leaf N concentrations and delta(13)C ratios, and stem water potential were measured for two growing seasons. Percent seed germination was consistently higher in open-canopy microhabitats and lowest at northern and inland sites. T. sebifera grew in all open-canopy microhabitats, even 300-500 km beyond its current distribution. Plant growth in closed-canopy habitats was lower, attributable to lower carbon gain per unit leaf area in shaded compared with open-canopy environments, especially at northern and inland sites. Neither competition, other than canopy shade, nor grazing was a key constraint on distribution at any scale. Our results demonstrate that T. sebifera is dispersal limited at landscape scales but limited locally by dispersal and overstory shade; it has yet to occupy the full extent of its new range in North America. Quantifying environmental factors both within and well beyond a species' current range can effectively highlight the limits on its distribution.


The role of the phosphatidylinositol 3-kinase (PI3K) pathway in the development and treatment of uterine cancer.
Uterine cancer is the most common gynecologic malignancy in the United States. Although surgery is often curative for women diagnosed in the early stages, prognosis for patients with advanced disease is poor. Alterations in the phosphatidylinositol 3-kinase (PI3K) pathway are known to play a significant role in the development of uterine cancer and provide a possible target for new therapies. PubMed was searched for articles of relevance to uterine cancer and the PI3K pathway. In addition, abstracts from key oncology congresses were scanned for data on novel therapeutic agents targeting the PI3K pathway. The PI3K pathway is an important promoter of cellular growth, metabolism, differentiation, proliferation, survival, and angiogenesis. It is often upregulated in uterine cancer, with the most frequent genetic alterations occurring in phosphatase and tensin homolog (PTEN), PIK3CA, and KRAS. Deregulation of the pathway has been associated with resistance to hormonal therapy and chemotherapy. Inhibitors of the PI3K pathway are in clinical testing in patients with solid tumors, including uterine cancer. Results with monotherapy demonstrate some clinical responses, but mainly as prolonged stable disease. PI3K pathway inhibitors are currently being evaluated in patients with tumors in which the PI3K pathway is deregulated. Another strategy being evaluated is the ability of PI3K pathway inhibitors to restore sensitivity to standard therapy. Investigational PI3K pathway inhibitors represent a promising new therapeutic strategy in uterine cancer. Exploration of effective drug combinations and their applicability to individual tumors will be important in the future clinical development of these agents.


Effect of HDL, from Japanese white rabbit administered a new cholesteryl ester transfer protein inhibitor JTT-705, on cholesteryl ester accumulation induced by acetylated low density lipoprotein in J774 macrophage.
We have previously reported a potent and specific cholesteryl ester transfer protein (CETP) inhibitor JTT-705 was a potentially anti-atherogenic compound (Nature 406 (2000) 203). In the present study, we investigated in vitro how this compound affects properties of high density lipoprotein (HDL) in Japanese white (JW) rabbits in terms of reverse cholesterol transport in J774 macrophages. Plasma HDL-cholesterol (C) level was significantly higher in the rabbits administered JTT-705 than in control rabbits on days 3 and 7. Both HDL(2) and HDL(3)-C levels were also significantly higher in JTT-705-administered rabbits than in control rabbits. During this period, plasma CETP activity was kept lower in JTT-705-administered rabbits than in controls. To determine how this compound affects the property of HDL particles, we investigated the C efflux induced by HDL from JTT-705-administered and control rabbits in J774 macrophages. Cholesterol ester (CE) concentration in J774 macrophages was reduced in proportion with increasing concentration of the added HDL to the culture media for J774 macrophages in both groups, suggesting that the HDL from JTT-705-administered rabbits was able to reduce CE concentration in J774 macrophages as efficiently as that from control rabbits. This result, together with the finding that the absolute HDL concentration increased in JW rabbits administered this CETP inhibitor, suggests that treatment with this new compound causes a beneficial effect on lipid metabolism in terms of anti-atherogenicity.


Estimate of anti-Plasmodium falciparum sporozoite activity in humans vaccinated with synthetic circumsporozoite protein (NANP)3.
A mathematical model was defined to estimate the degree of in vivo activity against Plasmodium falciparum sporozoites expressed by volunteers vaccinated with a synthetic peptide comprising the immunodominant epitope of the circumsporozoite protein. Relative to the course of infection in non-immunized controls, infections in vaccinated volunteers corresponded to the neutralization or delay of development of greater than 99% of challenge sporozoites.


Postoperative cage migration and subsidence following TLIF surgery is not associated with bony fusion.
Pseudarthrosis following transforaminal interbody fusion (TLIF) is not infrequent. Although cage migration and subsidence are commonly regarded as evidence of the absence of solid fusion, there is still no evidence of the influence of cage migration and subsidence on fusion. This study aimed to evaluate cage migration and subsidence using computed tomography (CT) DICOM data following lumbar interbody fusion. The effects of cage migration and subsidence on fusion and clinical outcomes were also assessed. A postoperative CT data set of 67 patients treated with monosegmental TLIF was analyzed in terms of cage position. To assess the effects of cage migration and subsidence on fusion, 12-month postoperative CT scans were used to assess fusion status. Clinical evaluation included the visual analog scale for pain and the Oswestry Disability Index. Postoperative cage migration occurred in 85.1% of all patients, and cage subsidence was observed in 58.2%. Radiological signs of pseudarthrosis was observed in 7.5% of the patients Neither cage migration nor subsidence affected the clinical or radiographic outcomes. No correlation was found between clinical and radiographic outcomes. The incidence of cage migration was considerable. However, as cage migration and subsidence were not associated with bony fusion, their clinical significance was considered limited.