Convex optimization of coincidence time resolution for a high-resolution PET system.
We are developing a dual panel breast-dedicated positron emission tomography (PET) system using LSO scintillators coupled to position sensitive avalanche photodiodes (PSAPD). The charge output is amplified and read using NOVA RENA-3 ASICs. This paper shows that the coincidence timing resolution of the RENA-3 ASIC can be improved using certain list-mode calibrations. We treat the calibration problem as a convex optimization problem and use the RENA-3's analog-based timing system to correct the measured data for time dispersion effects from correlated noise, PSAPD signal delays and varying signal amplitudes. The direct solution to the optimization problem involves a matrix inversion that grows order (n(3)) with the number of parameters. An iterative method using single-coordinate descent to approximate the inversion grows order (n). The inversion does not need to run to convergence, since any gains at high iteration number will be low compared to noise amplification. The system calibration method is demonstrated with measured pulser data as well as with two LSO-PSAPD detectors in electronic coincidence. After applying the algorithm, the 511 keV photopeak paired coincidence time resolution from the LSO-PSAPD detectors under study improved by 57%, from the raw value of 16.3 ±0.07 ns full-width at half-maximum (FWHM) to 6.92 ±0.02 ns FWHM ( 11.52 ±0.05 ns to 4.89 ±0.02 ns for unpaired photons).


Reassessment of (90)Sr, (137)Cs, and (134)Cs in the Coast off Japan Derived from the Fukushima Dai-ichi Nuclear Accident.
The years following the Fukushima Dai-ichi nuclear power plant (FDNPP) accident, the distribution of (90)Sr in seawater in the coast off Japan has received limited attention. However, (90)Sr is a major contaminant in waters accumulated within the nuclear facility and in the storage tanks. Seawater samples collected off the FDNPP in September 2013 showed radioactive levels significantly higher than pre-Fukushima levels within 6 km off the FDNPP. These samples, with up to 8.9 ± 0.4 Bq·m(-3) for (90)Sr, 124 ± 3 Bq·m(-3) for (137)Cs, and 54 ± 1 Bq·m(-3) for (134)Cs, appear to be influenced by ongoing releases from the FDNPP, with a characteristic (137)Cs/(90)Sr activity ratio of 3.5 ± 0.2. Beach surface water and groundwater collected in Sendai Bay had (137)Cs concentrations of up to 43 ± 1 Bq·m(-3), while (90)Sr was close to pre-Fukushima levels (1-2 Bq·m(-3)). These samples appear to be influenced by freshwater inputs carrying a (137)Cs/(90)Sr activity ratio closer to that of the FDNPP fallout deposited on land in the spring of 2011. Ongoing inputs of (90)Sr from FDNPP releases would be on the order of 2.3-8.5 GBq·d(-1) in September 2013, likely exceeding river inputs by 2-3 orders of magnitude. These results strongly suggest that a continuous surveillance of artificial radionuclides in the Pacific Ocean is still required.


Balance features in Alzheimer's disease and amnestic mild cognitive impairment.
We evaluated alterations of balance by stabilometry in patients with amnestic mild cognitive impairment (aMCI) and with mild-moderate Alzheimer's disease (AD). Fifteen patients with aMCI and 15 with mild AD were recruited according to the current diagnostic criteria. Fifteen healthy subjects of the same age range were recruited as controls. Stabilometry was carried out using a commercial 4 load cell platform. Statistical analysis of between group differences was performed using one-way analysis of variance for parametric data and Kruskal-Wallis tests for non-parametric data. Spearman correlation coefficients were used to investigate the association between cognitive test scores and stabilometric data. All stabilometry measures were significantly altered in mild AD patients compared to normal controls. Antero-posterior sway was found to be the most sensitive parameter, since it correlated with the ADAS-cog orientation subscale in AD patients, and also discriminated between aMCI and normal controls. Our study shows that impairment in balance is a feature not only of AD, but also of aMCI. The alterations found suggest that a progressive failure of the vestibular system, possibly linked to reduced hippocampal performance, may be responsible for such a feature. Further research must be focused on studying the predictive value of stabilometry in the conversion of aMCI.


Targeting Mitochondrial Structure Sensitizes Acute Myeloid Leukemia to Venetoclax Treatment.
The BCL2 family plays important roles in acute myeloid leukemia (AML). Venetoclax, a selective BCL2 inhibitor, has received FDA approval for the treatment of AML. However, drug resistance ensues after prolonged treatment, highlighting the need for a greater understanding of the underlying mechanisms. Using a genome-wide CRISPR/Cas9 screen in human AML, we identified genes whose inactivation sensitizes AML blasts to venetoclax. Genes involved in mitochondrial organization and function were significantly depleted throughout our screen, including the mitochondrial chaperonin CLPB. We demonstrated that CLPB is upregulated in human AML, it is further induced upon acquisition of venetoclax resistance, and its ablation sensitizes AML to venetoclax. Mechanistically, CLPB maintains the mitochondrial cristae structure via its interaction with the cristae-shaping protein OPA1, whereas its loss promotes apoptosis by inducing cristae remodeling and mitochondrial stress responses. Overall, our data suggest that targeting mitochondrial architecture may provide a promising approach to circumvent venetoclax resistance. SIGNIFICANCE: A genome-wide CRISPR/Cas9 screen reveals genes involved in mitochondrial biological processes participate in the acquisition of venetoclax resistance. Loss of the mitochondrial protein CLPB leads to structural and functional defects of mitochondria, hence sensitizing AML cells to apoptosis. Targeting CLPB synergizes with venetoclax and the venetoclax/azacitidine combination in AML in a p53-independent manner.See related commentary by Savona and Rathmell, p. 831.This article is highlighted in the In This Issue feature, p. 813.


Generative Deep Neural Networks for Inverse Materials Design Using Backpropagation and Active Learning.
In recent years, machine learning (ML) techniques are seen to be promising tools to discover and design novel materials. However, the lack of robust inverse design approaches to identify promising candidate materials without exploring the entire design space causes a fundamental bottleneck. A general-purpose inverse design approach is presented using generative inverse design networks. This ML-based inverse design approach uses backpropagation to calculate the analytical gradients of an objective function with respect to design variables. This inverse design approach is capable of overcoming local minima traps by using backpropagation to provide rapid calculations of gradient information and running millions of optimizations with different initial values. Furthermore, an active learning strategy is adopted in the inverse design approach to improve the performance of candidate materials and reduce the amount of training data needed to do so. Compared to passive learning, the active learning strategy is capable of generating better designs and reducing the amount of training data by at least an order-of-magnitude in the case study on composite materials. The inverse design approach is compared with conventional gradient-based topology optimization and gradient-free genetic algorithms and the pros and cons of each method are discussed when applied to materials discovery and design problems.


Combined immune checkpoint blockade as a therapeutic strategy for BRCA1-mutated breast cancer.
Immune checkpoint inhibitors have emerged as a potent new class of anticancer therapy. They have changed the treatment landscape for a range of tumors, particularly those with a high mutational load. To date, however, modest results have been observed in breast cancer, where tumors are rarely hypermutated. Because BRCA1-associated tumors frequently exhibit a triple-negative phenotype with extensive lymphocyte infiltration, we explored their mutational load, immune profile, and response to checkpoint inhibition in a Brca1-deficient tumor model. BRCA1-mutated triple-negative breast cancers (TNBCs) exhibited an increased somatic mutational load and greater numbers of tumor-infiltrating lymphocytes, with increased expression of immunomodulatory genes including PDCD1 (PD-1) and CTLA4, when compared to TNBCs from BRCA1-wild-type patients. Cisplatin treatment combined with dual anti-programmed death-1 and anti-cytotoxic T lymphocyte-associated antigen 4 therapy substantially augmented antitumor immunity in Brca1-deficient mice, resulting in an avid systemic and intratumoral immune response. This response involved enhanced dendritic cell activation, reduced suppressive FOXP3+ regulatory T cells, and concomitant increase in the activation of tumor-infiltrating cytotoxic CD8+ and CD4+ T cells, characterized by the induction of polyfunctional cytokine-producing T cells. Dual (but not single) checkpoint blockade together with cisplatin profoundly attenuated the growth of Brca1-deficient tumors in vivo and improved survival. These findings provide a rationale for clinical studies of combined immune checkpoint blockade in BRCA1-associated TNBC.


Adsorption and dissociation of NO on stepped Pt (533).
We present an experimental and theoretical investigation of the adsorption, desorption, and dissociation of NO on the stepped Pt (533) surface. By combining temperature programmed desorption and reflection absorption infrared spectroscopy, information about the adsorption sites at different temperatures is obtained. Surprisingly, metastable adsorption structures of NO can be produced through variation of the dosing temperature. We also show that part of the NO molecules adsorbed on the step sites dissociates around 450 K. After dissociation the N atoms can desorb either by combining with an O fragment, or with another N atom, resulting in NO and N(2). The N(2) production can be enhanced by coadsorbing CO on the surface: CO scavenges the oxygen atom, thereby suppressing associative recombinative desorption of N and O atoms. Density functional theory calculations are used to reveal the adsorption energies and vibrational frequencies of adsorbed NO as well as barriers for dissociation of NO and for diffusion of N atoms. The combined experimental results and theoretical calculations reveal that dissociation of NO is the rate limiting step in the formation of N(2).


The role of the midcingulate cortex in monitoring others' decisions.
A plethora of research has implicated the cingulate cortex in the processing of social information (i.e., processing elicited by, about, and directed toward others) and reward-related information that guides decision-making. However, it is often overlooked that there is variability in the cytoarchitectonic properties and anatomical connections across the cingulate cortex, which is indicative of functional variability. Here we review evidence from lesion, single-unit recording and functional imaging studies. Taken together, these support the claim that the processing of information that has the greatest influence on social behavior can be localized to the gyral surface of the midcingulate cortex (MCCg). We propose that the MCCg is engaged when predicting and monitoring the outcomes of decisions during social interactions. In particular, the MCCg processes statistical information that tracks the extent to which the outcomes of decisions meet goals when interacting with others. We provide a novel framework for the computational mechanisms that underpin such social information processing in the MCCg. This framework provides testable hypotheses for the social deficits displayed in autism spectrum disorders and psychopathy.


Multiple repeat prostate biopsies and the detection of clinically insignificant cancer in men with large prostates.
To determine the impact of repeating prostate biopsies on the risk of detecting clinically insignificant prostate cancer (PCa) in larger prostate glands. We performed a retrospective cohort study using patients enrolled in our institutional PCa registry from 1991 to 2008 to assess the association of prostate volume and clinically insignificant PCa in men undergoing multiple prostate biopsies. Patients were stratified by prostate volume into 2 cohorts (<50 cm(3) or ≥50 cm(3)). Additionally, patients were stratified by prostate biopsy on which PCa was identified (1 biopsy or ≥3 biopsies). Within the subgroup of patients with prostate volume ≥50 cm(3) requiring ≥3 biopsies before cancer diagnosis, 72.6% (45/62) had pathologic Gleason scores ≤6 and 81.6% (49/60) had an estimated tumor volume of ≤10% at the time of radical prostatectomy. This was significantly different from patients with prostate volume <50 cm(3) diagnosed on their first biopsy, in which only 48.5% (656/1349) were found to have Gleason scores ≤6 and 54.2% (705/1300) had estimated tumor volume ≤10% (P <.01). There was no significant difference in the rate of Gleason score upgrading at time of prostatectomy between any of the subgroups. PCas detected in men with prostatic enlargement requiring multiple biopsies are more likely to be low-grade, low-volume tumors at final pathology than men without prostate enlargement. Men with larger prostates who have already had prior negative biopsies should be counseled regarding the increased risk of detecting clinically insignificant PCa with additional biopsies.


Dietary fiber intake reduces risk for colorectal adenoma: a meta-analysis.
Reports on the association between dietary fiber intake and risk of colorectal adenoma (CRA), the precursor of colorectal cancer, have been inconsistent. We conducted a meta-analysis of case-control and cohort studies to analyze this association. We searched the MEDLINE and EMBASE databases to identify relevant studies published through July 2013. A random-effects model was used to estimate summary relative risks (SRRs) and 95% confidence intervals (CIs) for associations between fiber intake and CRA risk. Heterogeneity among studies was assessed using the Cochran Q and I(2) statistics. Our meta-analysis included 20 studies involving 10,948 subjects with CRA. The SRRs of CRA for total dietary fiber were 0.72 (95% CI, 0.63-0.83) in a high- vs low-intake analysis and 0.91 (95% CI, 0.87-0.95) per 10-g/day increase in fiber intake in a dose-response model. Subgroup analyses indicated a significant inverse association between total fiber intake and CRA risk in case-control studies (SRR, 0.66; 95% CI, 0.56-0.77), but not in cohort studies (SRR, 0.92; 95% CI, 0.76-1.10). The SRRs of CRA were 0.84 for fruit fiber (95% CI, 0.76-0.94; n = 6 studies), 0.93 for vegetable fiber (95% CI, 0.84-1.04; n = 6 studies), and 0.76 for cereal fiber (95% CI, 0.62-0.92; n = 9 studies) in high- vs low-intake analyses. Our findings support the hypothesis that high dietary fiber intake is associated inversely with CRA risk. Further studies with prospective designs that use validated questionnaires and control for important confounders are warranted.


Regulatory control and the costs and benefits of biochemical noise.
Experiments in recent years have vividly demonstrated that gene expression can be highly stochastic. How protein concentration fluctuations affect the growth rate of a population of cells is, however, a wide-open question. We present a mathematical model that makes it possible to quantify the effect of protein concentration fluctuations on the growth rate of a population of genetically identical cells. The model predicts that the population's growth rate depends on how the growth rate of a single cell varies with protein concentration, the variance of the protein concentration fluctuations, and the correlation time of these fluctuations. The model also predicts that when the average concentration of a protein is close to the value that maximizes the growth rate, fluctuations in its concentration always reduce the growth rate. However, when the average protein concentration deviates sufficiently from the optimal level, fluctuations can enhance the growth rate of the population, even when the growth rate of a cell depends linearly on the protein concentration. The model also shows that the ensemble or population average of a quantity, such as the average protein expression level or its variance, is in general not equal to its time average as obtained from tracing a single cell and its descendants. We apply our model to perform a cost-benefit analysis of gene regulatory control. Our analysis predicts that the optimal expression level of a gene regulatory protein is determined by the trade-off between the cost of synthesizing the regulatory protein and the benefit of minimizing the fluctuations in the expression of its target gene. We discuss possible experiments that could test our predictions.


Mobility closure in the upper class: assessing time and forms of capital.
Most understandings of the ways classes become social groupings centre on processes of mobility closure whereby mutual appreciation and recognition within classes arise from homogenous experiences over time. The mapping of such structured biographies, however, remains understudied. This paper explores intra- and intergenerational mobility patterns in the upper strata of the Norwegian class structure and aims to include temporal processes and multiple forms of capital in the quantification of class trajectories. By combining multiple correspondence analysis and social sequence analysis, two important but often neglected aspects of recruitment to the upper class are emphasized: first, by introducing multiple forms of capital, different ways of maintaining mobility closure are demonstrated; second, different pathways to power are highlighted by distinguishing between divergent class careers. A key aim of the analysis is to explore internal divisions within the upper class in forms of parental capital (an 'origin space') and link these divisions to a typology of 'destination careers' in adulthood. The analysis suggests that individuals from modest origins are more likely to have careers that feature a biographically late arrival and/or short-term affiliations to upper-class positions whereas individuals from families rich in capital are more likely to have stable careers in the upper-class fractions from which they originate. The analysis thus reveals important divisions in the trajectories of Norwegians who reach the upper class; not only are there differences in their upbringing in terms of the availability of different amounts and types of capital but such divisions also seem linked to their own class careers later in adulthood.


Facing one's implicit biases: From awareness to acknowledgment.
Expanding on conflicting theoretical conceptualizations of implicit bias, 6 studies tested the effectiveness of different procedures to increase acknowledgment of harboring biases against minorities. Participants who predicted their responses toward pictures of various minority groups on future implicit association tests (IATs) showed increased alignment between implicit and explicit preferences (Studies 1-3), greater levels of explicit bias (Studies 1-3), and increased self-reported acknowledgment of being racially biased (Studies 4-6). In all studies, effects of IAT score prediction were significant even when participants did not actually complete IATs. Effects of predicting IAT scores were moderated by nonprejudicial goals, in that IAT score prediction increased acknowledgment of bias for participants with strong nonprejudicial goals, but not for participants with weak nonprejudicial goals (Study 4). Mere completion of IATs and feedback on IAT performance had inconsistent effects across studies and criterion measures. Instructions to attend to one's spontaneous affective reactions toward minority group members increased acknowledgment of bias to the same extent as IAT score prediction (Study 6). The findings are consistent with conceptualizations suggesting that (a) implicit evaluations are consciously experienced as spontaneous affective reactions and (b) directing people's attention to their spontaneous affective reactions can increase acknowledgment of bias. Implications for theoretical conceptualizations of implicit bias and interventions that aim to reduce discrimination via increased acknowledgment of bias are discussed. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


Diurnal cortisol rhythm as a predictor of lung cancer survival.
Poorly coordinated diurnal cortisol and circadian rest-activity rhythms predict earlier mortality in metastatic breast and colorectal cancer, respectively. We examined the prognostic value of the diurnal cortisol rhythm in lung cancer. Lung cancer patients (n=62, 34 female) were within 5 years of diagnosis and had primarily non small-cell lung cancer, with disease stage ranging from early to advanced. Saliva collected over two days allowed calculation of the diurnal cortisol slope and the cortisol awakening response (CAR). Lymphocyte numbers and subsets were measured by flow cytometry. Survival data were obtained for 57 patients. Cox Proportional Hazards analyses were used to test the prognostic value of the diurnal cortisol rhythm on survival calculated both from study entry and from initial diagnosis. The diurnal cortisol slope predicted subsequent survival over three years. Early mortality occurred among patients with higher slopes, or relatively "flat" rhythms indicating lack of normal diurnal variation (Cox Proportional Hazards p=.009). Cortisol slope also predicted survival time from initial diagnosis (p=.012). Flattened profiles were linked with male gender (t=2.04, df=59, p=.046) and low total and cytotoxic T cell lymphocyte counts (r=-.39 and -.30, p=.004 and .035, respectively). After adjustment for possible confounding factors, diurnal slope remained a significant, independent predictor of survival. Flattening of the diurnal cortisol rhythm predicts early lung cancer death. Data contribute to growing evidence that circadian disruption accelerates tumor progression.


Selenium-enriched Spirulina protects INS-1E pancreatic beta cells from human islet amyloid polypeptide-induced apoptosis through suppression of ROS-mediated mitochondrial dysfunction and PI3/AKT pathway.
Human islet amyloid polypeptide (hIAPP) aggregation is linked to loss of pancreatic beta cells in type 2 diabetes, in part due to oxidative stress. Currently, little is known about the effects of selenium-enriched Spirulina on beta cells with the presence of hIAPP. In this study, INS-1E rat insulinoma cells were used as a model to evaluate in vitro protective effects of Se-enriched Spirulina extract (Se-SE) against hIAPP-induced cell death, as well as the underlying mechanisms. Flow cytometric analysis was used to evaluate cell apoptosis, mitochondrial membrane potential (ΔΨm) and ROS generation. Caspase activity was measured using a fluorometric method. Western blotting was applied to detect protein expression. Our results showed that exposure of INS-1E cells to hIAPP resulted in cell viability loss, LDH release and appearance of sub-G peak. However, cytotoxicity of hIAPP was significantly attenuated by co-treatment with Se-SE. Se-SE also inhibited hIAPP-induced activation of caspase-3, -8 and -9. Additionally, hIAPP-induced accumulation of ROS and superoxide was suppressed by co-treatment with Se-SE. Moreover, Se-SE was able to prevent hIAPP-induced depletion of ΔΨm and intracellular ATP, reduction in mitochondrial mass, changes in the expression of Bcl-2 family members, release of mitochondrial apoptogenic factors. Furthermore, hIAPP-mediated AKT inhibition was restored by co-treatment with Se-SE. Our results showed that Se-SE protects INS-1E cells from hIAPP-induced cell death through preventing ROS overproduction, mitochondrial dysfunction and modulating PI3K/AKT pathway.


Modelling the interdependence between the stoichiometry of receptor oligomerization and ligand binding for a coexisting dimer/tetramer receptor system.
Many G protein-coupled receptors have been shown to exist as oligomers, but the oligomerization state and the effects of this on receptor function are unclear. For some G protein-coupled receptors, in ligand binding assays, different radioligands provide different maximal binding capacities. Here we have developed mathematical models for co-expressed dimeric and tetrameric species of receptors. We have considered models where the dimers and tetramers are in equilibrium and where they do not interconvert and we have also considered the potential influence of the ligands on the degree of oligomerization. By analogy with agonist efficacy, we have considered ligands that promote, inhibit or have no effect on oligomerization. Cell surface receptor expression and the intrinsic capacity of receptors to oligomerize are quantitative parameters of the equations. The models can account for differences in the maximal binding capacities of radioligands in different preparations of receptors and provide a conceptual framework for simulation and data fitting in complex oligomeric receptor situations.


Treadmill exercise ameliorates symptoms of Alzheimer disease through suppressing microglial activation-induced apoptosis in rats.
Alzheimer disease (AD) is a most common form of dementia and eventually causes impairments of learning ability and memory function. In the present study, we investigated the effects of treadmill exercise on the symptoms of AD focusing on the microglial activation-induced apoptosis. AD was made by bilateral intracerebroventricular injection of streptozotocin. The rats in the exercise groups were made to run on a treadmill once a day for 30 min during 4 weeks. The distance and latency in the Morris water maze task and the latency in the step-down avoidance task were increased in the AD rats, in contrast, treadmill exercise shortened these parameters. The numbers of terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling-positive and caspase-3-positive cells in the hippocampal dentate gyrus were decreased in the AD rats, in contrast, treadmill exercise suppressed these numbers. Expressions of glial fibrillary acidic protein (GFAP) and cluster of differentiation molecule 11B (CD11b) in the hippocampal dentate gyrus were increased in the AD rats, in contrast, treadmill exercise suppressed GFAP and CD11b expressions. Bax expression was increased and Bcl-2 expression was decreased in the hippocampus of AD rats, in contrast, treadmill exercise decreased Bax expression and increased Bcl-2 expression. The present results demonstrated that treadmill exercise ameliorated AD-induced impairments of spatial learning ability and short-term memory through suppressing apoptosis. The antiapoptotic effect of treadmill exercise might be ascribed to the inhibitory effect of treadmill exercise on microglial activation.


Origin and diversification of the human parasite Schistosoma mansoni.
Schistosoma mansoni is the most widespread of the human-infecting schistosomes, present in 54 countries, predominantly in Africa, but also in Madagascar, the Arabian Peninsula, and the Neotropics. Adult-stage parasites that infect humans are also occasionally recovered from baboons, rodents, and other mammals. Larval stages of the parasite are dependent upon certain species of freshwater snails in the genus Biomphalaria, which largely determine the parasite's geographical range. How S. mansoni genetic diversity is distributed geographically and among isolates using different hosts has never been examined with DNA sequence data. Here we describe the global phylogeography of S. mansoni using more than 2500 bp of mitochondrial DNA (mtDNA) from 143 parasites collected in 53 geographically widespread localities. Considerable within-species mtDNA diversity was found, with 85 unique haplotypes grouping into five distinct lineages. Geographical separation, and not host use, appears to be the most important factor in the diversification of the parasite. East African specimens showed a remarkable amount of variation, comprising three clades and basal members of a fourth, strongly suggesting an East African origin for the parasite 0.30-0.43 million years ago, a time frame that follows the arrival of its snail host. Less but still substantial variation was found in the rest of Africa. A recent colonization of the New World is supported by finding only seven closely related New World haplotypes which have West African affinities. All Brazilian isolates have nearly identical mtDNA haplotypes, suggesting a founder effect from the establishment and spread of the parasite in this large country.


Insulin-like growth factors decrease oxygen-regulated erythropoietin production by human hepatoma cells (Hep G2).
We examined the effects of insulin-like growth factors (IGFs) and insulin on erythropoietin (EPO) production by human hepatoma cells (Hep G2). Compared with normoxia (20% O2), EPO production by Hep G2 cells during a 72-h incubation was stimulated fivefold by exposure to low oxygen tension (1% O2) and nearly threefold by exposure to cobalt chloride (100 microM). IGF-I caused a concentration-dependent attenuation of EPO formation under normoxic conditions and inhibited (maximally 50%) EPO production stimulated by either low oxygen tension or cobalt [half-maximal effect (ED50) approximately 5 nM]. The increase of EPO mRNA levels in response to hypoxia was significantly reduced by IGF-I. Similarly to IGF-I, IGF-II (ED50 approximately 8 nM) and insulin (ED50 approximately 80 nM) also inhibited EPO formation in Hep G2 cells. IGF-I (100 pM-100 nM) stimulated the incorporation of radiolabeled alanine as a measure for total protein synthesis, 3H-labeled thymidine incorporation into DNA, and glycogen synthesis at 20 and 1% O2 in a concentration-dependent fashion. IGF-I exhibited a high affinity for the IGF-I receptor (apparent Kd approximately 3 nM). Unlabeled insulin was greater than 100-fold less potent than IGF-I in competing for 125I-IGF-I binding (apparent Kd approximately 360 nM). Conversely, insulin bound to the insulin receptor with high affinity (apparent Kd approximately 0.3 nM), whereas IGF-I was less than 1% as potent in competing for 125I-insulin binding. In summary, IGFs and insulin exert a negative control function on oxygen-regulated EPO production in Hep G2 cells. The inhibitory effect of IGFs and insulin on EPO formation appears to be mediated via the IGF-I receptor.


Induction of the cytokine signal regulator SOCS3/CIS3 as a therapeutic strategy for treating inflammatory arthritis.
Immune and inflammatory systems are controlled by multiple cytokines, including ILs and INFs. These cytokines exert their biological functions through Janus tyrosine kinases and STAT transcription factors. One such cytokine, IL-6, has been proposed to contribute to the development of rheumatoid arthritis (RA). We found that STAT3 was strongly tyrosine phosphorylated in synovial tissue of RA patients, but not those with osteoarthritis. Blockade of the IL-6-gp130-JAK-STAT3-signaling pathway might therefore be beneficial in the treatment of RA. We show here that the mRNA for the endogenous cytokine signaling repressor CIS3/SOCS3 is abundantly expressed in RA patients. To determine whether CIS3 is effective in treating experimental arthritis, a recombinant adenovirus carrying the CIS3 cDNA was injected periarticularly into the ankle joints of mice with antigen-induced arthritis or collagen-induced arthritis (CIA). Periarticular injection of CIS3 adenovirus drastically reduced the severity of arthritis and joint swelling compared with control groups. CIS3 was more effective than a dominant-negative form of STAT3 in the CIA model. Thus, induction of CIS3 could represent a new approach for effective treatment of RA.


The longest tumor diameter in one dimension as a predictor for skeletal metastasis in renal cell carcinoma.
Renal cell carcinoma (RCC) comprises a diverse group of malignant neoplasms that have multifarious histopathological features and biological behavior. One-third of RCC patients develops skeletal metastasis with a poor 5-year survival rate. Data explaining how some of these tumors show sooner bony metastasis than expected is sparse. The objective of this study was to identify whether tumor size can act as a predictor of bony metastases among patients of RCC. We retrospectively reviewed contrast enhanced computed tomography (CECT) scan and clinical records of 66 patients with RCC, who fulfilled specified inclusion criteria. Patients who had bony metastasis at the time of presentation were selected as case and those without skeletal metastasis were referred to as controls. Receiver operating characteristic (ROC) curve analysis was used to determine the appropriate cut-off value for tumor size, which was measured as the longest tumor diameter (LTD) in one-dimensional (1D). Of the 66 patients selected, 30% developed bone metastasis. The tumor size of RCCs significantly correlated with the presence of skeletal metastasis in our study. None of the patients with 1D LTD <4.8 cm on CECT were found to have skeletal metastasis. ROC analysis revealed that the accuracy of the LTD in predicting bone metastasis was high with an area under ROC curve of 0.823. A cut-off value of 7.5 cm had a sensitivity of 78.9% and specificity of 80.9%. The 1D LTD with a cut-off value of 7.5 cm, at the time of presentation is an important predictor of skeletal metastasis. The result of this study may have role in triage of patients into a subgroup which mandates more aggressive treatment and monitoring.


The Versatility of 980 nm Diode Laser in Dentistry: A Case Series.
Introduction: Laser surgery has been considered a popular alternative over conventional modalities in dentistry during the last few years. Among different types of lasers, diode lasers have gained special attention in oral soft tissue surgery. Case Reports: Five patients were referred to a private office. After careful evaluation of medical history and oral examination, oral diagnosis and treatment plan of each patient was established as follows: (1) A 21-year-old female with ankyloglossia (tongue-tie); (2) A 65-year-old female with a poor denture fit needing vestibuloplasty and frenectomy; (3) A 10-year-old male patient with pigmented gingiva in mandible and maxilla; (4) A 14-year-old female needing exposure of maxillary right canine for bracket bonding; and (5) A 25-year-old female patient who has a gingival maxillary frenum with a nodule. The treatment plan for all the patients was laser surgery with diode laser at 980 nm, in continuous mode. Results: All the patients experienced normal healing process with no postoperative complications. Favorable outcomes of laser surgery were observed on follow-up sessions. Conclusion: Considering the versatility of the 980 nm diode laser in oral soft tissue surgeries and the advantages of laser surgery, this study suggests the use of 980 nm diode laser in this regard.


Inhibition of AMPK-associated autophagy enhances caffeic acid phenethyl ester-induced cell death in C6 glioma cells.
An increasing number of studies show that AMP-activated protein kinase (AMPK) activation can inhibit apoptosis. To clarify the antitumor mechanism of caffeic acid phenethyl ester (CAPE) and achieve increased therapeutic efficiency, we investigated the potential roles of AMPK and autophagy in CAPE treatment against C6 glioma cells. The roles of AMPK and autophagy inhibition in CAPE's cytotoxic action were investigated. Phosphorylation of AMPK and mitogen-activated protein kinases (MAPKs) were observed in tumor cells following CAPE treatment. A combination of CAPE and the AMPK inhibitor, compound C, resulted in augmented cell death. Similar effects of compound C were observed in response to changes in the mitochondrial membrane potential ( ΔΨ(m)). Small interfering RNA-mediated AMPK downregulation increased CAPE-induced cell death. The results suggest that AMPK activation plays a role in diminishing apoptosis. CAPE treatment induced an increase in LC3 conversion as represented by the LC3-II/LC3-I ratio. Enlarged lysosomes and autophagosomes were present according to electron microscopy. The autophagy inhibitor, 3-MA, caused increased CAPE cytotoxicity, which suggests that autophagy induction protected glioma cells from CAPE. The combination of CAPE with autophagy and AMPK inhibitors markedly enhanced the cytotoxicity toward C6 glioma cells. Accordingly, CAPE-triggered activation of AMPK and the autophagic response protected tumor cells from apoptotic death. This provides new insights for combined therapy to enhance the therapeutic potential of cancer treatments.


From coral reefs to Joshua trees: What ecological interactions teach us about the adaptive capacity of biodiversity in the Anthropocene.
The pervasive loss of biodiversity in the Anthropocene necessitates rapid assessments of ecosystems to understand how they will respond to anthropogenic environmental change. Many studies have sought to describe the adaptive capacity (AC) of individual species, a measure that encompasses a species' ability to respond and adapt to change. Only those adaptive mechanisms that can be used over the next few decades (e.g. via novel interactions, behavioural changes, hybridization, migration, etc.) are relevant to the timescale set by the rapid changes of the Anthropocene. The impacts of species loss cascade through ecosystems, yet few studies integrate the capacity of ecological networks to adapt to change with the ACs of its species. Here, we discuss three ecosystems and how their ecological networks impact the AC of species and vice versa. A more holistic perspective that considers the AC of species with respect to their ecological interactions and functions will provide more predictive power and a deeper understanding of what factors are most important to a species' survival. We contend that the AC of a species, combined with its role in ecosystem function and stability, must guide decisions in assigning 'risk' and triaging biodiversity loss in the Anthropocene. This article is part of the theme issue 'Ecological complexity and the biosphere: the next 30 years'.


Influence of CMV/EBV serostatus on respiratory infection incidence during 4 months of winter training in a student cohort of endurance athletes.
The purpose of this study was to examine the influence of previous infection with cytomegalovirus (CMV) or Epstein Barr virus (EBV) on the incidence, severity and duration of upper respiratory tract infections (URTIs) in endurance athletes during a 4-month winter training period. Blood samples were obtained from 236 subjects (166 males and 70 females, aged 18-35 years) at the start of the study period. Plasma samples were analysed for CMV and EBV serostatus. Weekly training and daily illness logs were kept for the next 16 weeks. With regard to CMV/EBV serostatus, the results indicated that athletes with previous CMV infection (n = 58, 25 % of cohort) had significantly fewer URTI symptom days (median 2 vs. 4 days, p = 0.033) during the study period than those with no previous infection (n = 178, 75 %), whereas positive EBV serostatus (n = 197, 84 %) had no influence on URTI episode incidence, severity or duration. Moreover, we found that athletes with prior infection of both CMV and EBV (n = 50, 21 %) had 50 % fewer URTI episodes (p = 0.04) and symptom days (median 2 vs 8 days, p = 0.01) than athletes who were seronegative for both CMV and EBV (n = 31, 13 %). Previous coinfection with CMV and EBV might promote protective immune surveillance to lower the risk of URTI. Further research is required to clarify why previous CMV and EBV infection reduces the incidence of URTI.


Representation of dynamic interaural phase difference in auditory cortex of awake rhesus macaques.
Neurons in auditory cortex of awake primates are selective for the spatial location of a sound source, yet the neural representation of the binaural cues that underlie this tuning remains undefined. We examined this representation in 283 single neurons across the low-frequency auditory core in alert macaques, trained to discriminate binaural cues for sound azimuth. In response to binaural beat stimuli, which mimic acoustic motion by modulating the relative phase of a tone at the two ears, these neurons robustly modulate their discharge rate in response to this directional cue. In accordance with prior studies, the preferred interaural phase difference (IPD) of these neurons typically corresponds to azimuthal locations contralateral to the recorded hemisphere. Whereas binaural beats evoke only transient discharges in anesthetized cortex, neurons in awake cortex respond throughout the IPD cycle. In this regard, responses are consistent with observations at earlier stations of the auditory pathway. Discharge rate is a band-pass function of the frequency of IPD modulation in most neurons (73%), but both discharge rate and temporal synchrony are independent of the direction of phase modulation. When subjected to a receiver operator characteristic analysis, the responses of individual neurons are insufficient to account for the perceptual acuity of these macaques in an IPD discrimination task, suggesting the need for neural pooling at the cortical level.


Local control following single-dose intraoperative radiotherapy prior to surgical excision of early-stage breast cancer.
Multiple partial breast radiotherapy techniques are available. We have previously presented the technical details of our procedure of delivering partial breast irradiation with a single fraction of intraoperative radiotherapy (IORT) targeting the tumor in situ prior to partial mastectomy. This study details our completed, single-institution trial. An IRB-approved, DSMB-monitored phase II trial was performed with the following inclusion criteria: women age ≥48, ultrasound-visible invasive ductal cancers <3 cm, clinically negative axillary nodes. IORT was delivered using mobile electron irradiator, at least a 1.5-cm radial and 1-cm deep margin; patients received 15 Gy and immediately underwent partial mastectomy. Ipsilateral breast recurrence was classified as true/marginal, elsewhere in the breast or nodal basin. Kaplan-Meier methods were used to estimate survival functions and exact 95% confidence intervals are reported. Between 2003 and 2007, 71 women underwent IORT (median follow-up: 3.5 years). For patients with tumor-involved or close margins, additional therapy was required: 7 patients, total mastectomy; 11, whole breast radiation. Four women experienced invasive ipsilateral breast failures (1 new primary, 3 margin recurrences) for a 3-year local control rate of 49 of 53 (94.8%; 95% confidence interval 92.4% [95% CI] 84.2–98.3%), actuarial three-year in breast recurrence was 8% (95% CI 2–18%), and breast cancer-specific survival was 100%. Intraoperative radiotherapy delivered to an in situ tumor is feasible, but our local control rate at 3.5 years is concerning. Possible changes to this technique to improve local control rates include better preoperative imaging (MRI), routine intraoperative ultrasound, and improved IORT delivery (larger cone, increased dose).


Vitamin B12 Supplementation in Diabetic Neuropathy: A 1-Year, Randomized, Double-Blind, Placebo-Controlled Trial.
To investigate the effect of normalizing vitamin B12 (B12) levels with oral B12 (methylcobalamin) 1000 μg/day for one year in patients with diabetic neuropathy (DN). In this prospective, double-blind, placebo-controlled trial, 90 patients with type 2 diabetes on metformin for at least four years and both peripheral and autonomic DN were randomized to an active treatment group (n = 44) receiving B12 and a control group (n = 46) receiving a placebo. All patients had B12 levels less than 400 pmol/L. Subjects underwent measurements of sural nerve conduction velocity (SNCV), sural nerve action potential (amplitude) (SNAP), and vibration perception threshold (VPT), and they performed cardiovascular autonomic reflex tests (CARTs: mean circular resultant (MCR), Valsalva test, postural index, and orthostatic hypotension). Sudomotor function was assessed with the SUDOSCAN that measures electrochemical skin conductance in hands and feet (ESCH and ESCF, respectively). We also used the Michigan Neuropathy Screening Instrument Questionnaire and Examination (MNSIQ and MNSIE, respectively) and questionnaires to evaluate quality of life (QoL) and level of pain (pain score). B12 levels increased from 232.0 ± 71.8 at baseline to 776.7 ± 242.3 pmol/L at follow-up, p < 0.0001, in the active group but not in the control group. VPT, MNSIQ, QoL, pain score, SNCV, SNAP, and ESCF significantly improved in the active group (p < 0.001, p = 0.002, p < 0.0001, p < 0.000, p < 0.0001, p < 0.0001, and p = 0.014, respectively), whereas CARTS and MNSIE improved but not significantly. MCR, MNSIQ, SNCV, SNAP, and pain score significantly deteriorated in the control group (p = 0.025, p = 0.017, p = 0.045, p < 0.0001, and p < 0.0001, respectively). The treatment of patients with DN with 1 mg of oral methylcobalamin for twelve months increased plasma B12 levels and improved all neurophysiological parameters, sudomotor function, pain score, and QoL, but it did not improve CARTS and MNSIE.


Enhanced radiation sensitivity in prostate cancer by gold-nanoparticles.
Nanotechnology is an emerging field with significant translational potential in medicine. In this study, we applied gold nanoparticles (GNP) to enhance radiation sensitivity and growth inhibition in radiation-resistant human prostate cancer cells. Gold nanoparticles (GNPs) were synthesized using HAuCl4 as the gold particle source and NaBH4 as the reductant. Either thio-glucose or sodium citrate was then added to the solution separately to bind the GNPs to form thio-glucose-capped gold nanoparticles (Glu-GNP) and neutral gold nanoparticles (TGS-GNPs). Human prostate carcinoma DU-145 cells were exposed to vehicle, irradiation, 15nM TGS-GNPs, or 15nM Glu-GNPs, or GNPs plus irradiation. The uptake assays of GNP were performed using hemocytometer to count cells and the mass spectrometry was applied to calculate gold mass. The cytotoxicity induced by GNPs, irradiation, or GNPs plus irradiation was measured using a standard colorimetric MTT assay. Exposure to Glu-GNPs resulted in a three times increase of nanoparticle uptake compared to that of TGS-GNPs in each target cell (p < 0.005). Cytoplasmic intracellular uptake of both TGS-GNPs and Glu-GNPs resulted in a growth inhibition by 30.57% and 45.97% respectively, comparing to 15.88% induced by irradiation alone, in prostate cancer cells after exposure to the irradiation. Glu-GNPs showed a greater enhancement, 1.5 to 2 fold increases within 72 hours, on irradiation cytotoxicity compared to TGS-GNPs. Tumour killing, however, did not appear to correlate linearly with nanoparticle uptake concentrations. These results showed that functional glucose-bound gold nanoparticles enhanced radiation sensitivity and toxicity in prostate cancer cells. In vivo studies will be followed to verify our research findings.


Constraints to implementing the Essential Health Package in Malawi.
Increasingly seen as a useful tool of health policy, Essential or Minimal Health Packages direct resources to interventions that aim to address the local burden of disease and be cost-effective. Less attention has been paid to the delivery mechanisms for such interventions. This study aimed to assess the degree to which the Essential Health Package (EHP) in Malawi was available to its population and what health system constraints impeded its full implementation. The first phase of this study comprised a survey of all facilities in three districts including interviews with all managers and clinical staff. In the second and third phase, results were discussed with District Health Management Teams and national level stakeholders, respectively, including representatives of the Ministry of Health, Central Medical Stores, donors and NGOs. The EHP in Malawi is focussing on the local burden of disease; however, key constraints to its successful implementation included a widespread shortage of staff due to vacancies but also caused by frequent trainings and meetings (only 48% of expected man days of clinical staff were available; training and meetings represented 57% of all absences in health centres). Despite the training, the percentage of health workers aware of vital diagnostic and therapeutic approaches to EHP conditions was weak. Another major constraint was shortages of vital drugs at all levels of facilities (e.g. Cotrimoxazole was sufficiently available to treat the average number of patients in only 27% of health centres). Although a few health workers noted some improvement in infrastructure and working conditions, they still considered them to be widely inadequate. In Malawi, as in similar resource poor countries, greater attention needs to be given to the health system constraints to delivering health care. Removal of these constraints should receive priority over the considerable focus on the development and implementation of essential packages of interventions.


MitoHiFi: a python pipeline for mitochondrial genome assembly from PacBio high fidelity reads.
PacBio high fidelity (HiFi) sequencing reads are both long (15-20 kb) and highly accurate (> Q20). Because of these properties, they have revolutionised genome assembly leading to more accurate and contiguous genomes. In eukaryotes the mitochondrial genome is sequenced alongside the nuclear genome often at very high coverage. A dedicated tool for mitochondrial genome assembly using HiFi reads is still missing. MitoHiFi was developed within the Darwin Tree of Life Project to assemble mitochondrial genomes from the HiFi reads generated for target species. The input for MitoHiFi is either the raw reads or the assembled contigs, and the tool outputs a mitochondrial genome sequence fasta file along with annotation of protein and RNA genes. Variants arising from heteroplasmy are assembled independently, and nuclear insertions of mitochondrial sequences are identified and not used in organellar genome assembly. MitoHiFi has been used to assemble 374 mitochondrial genomes (368 Metazoa and 6 Fungi species) for the Darwin Tree of Life Project, the Vertebrate Genomes Project and the Aquatic Symbiosis Genome Project. Inspection of 60 mitochondrial genomes assembled with MitoHiFi for species that already have reference sequences in public databases showed the widespread presence of previously unreported repeats. MitoHiFi is able to assemble mitochondrial genomes from a wide phylogenetic range of taxa from Pacbio HiFi data. MitoHiFi is written in python and is freely available on GitHub ( https://github.com/marcelauliano/MitoHiFi ). MitoHiFi is available with its dependencies as a Docker container on GitHub (ghcr.io/marcelauliano/mitohifi:master).


Effects of prophylactic and therapeutic antimicrobial uses in small-scale chicken flocks.
Antimicrobials are extensively used both prophylactically and therapeutically in poultry production. Despite this, there are little data on the effect of antimicrobial use (AMU) on disease incidence rate and per cent mortality. We investigated the relationships between AMU and disease and between AMU and mortality using data from a large (n = 322 flocks) cohort of small-scale chicken flocks in the Mekong Delta, Vietnam, that were followed longitudinally from day old to slaughter (5,566 observation weeks). We developed a parameterized algorithm to emulate a randomized control trial from observational data by categorizing the observation weeks into 'non-AMU', 'prophylactic AMU' and 'therapeutic AMU'. To evaluate the prophylactic AMU effect, we compared the frequencies of clinical signs in 'non-AMU' and 'prophylactic AMU' periods. To analyse therapeutic AMU, we compared weekly per cent mortality between the weeks of disease episodes before and after AMU. Analyses were stratified by clinical signs (4) and antimicrobial classes (13). Prophylactic AMU never reduced the probability of disease, and some antimicrobial classes such as lincosamides, amphenicols and penicillins increased the risk. The risk of diarrhoea consistently increased with prophylactic AMU. Therapeutic AMU often had an effect on mortality, but the pattern was inconsistent across the combinations of antimicrobial classes and clinical signs with 14/29 decreasing and 11/29 increasing the per cent weekly mortality. Lincosamides, methenamines and cephalosporins were the only three antimicrobial classes that always decreased the mortality when used therapeutically. Results were robust respective to the parameters values of the weeks categorization algorithm. This information should help support policy efforts and interventions aiming at reducing AMU in animal production.


Noise characterization in circular dichroism spectroscopy.
Circular dichroism (CD), defined as the difference in absorption between left and right circularly polarized light, is used to spectroscopically study the structures of chiral materials. In this article, various methodologies are presented for characterizing the performance of CD spectrometers to determine (1) experimental conditions for optimal data collection, (2) noise characteristics dependent on machine parameters, (3) the relative significance of spectral data as a function of detector gain, and (4) stray light and dark current as a function of wavelength. The results of case studies of two commercial CD spectrometers (specifically, Jasco J810 and J815) are described. The analyses show that the variation of CD signal is Poisson distributed and hence can be considered shot noise. Also, optimum scan parameters are established and a weighting function of CD data significance is produced so that wavelength-dependent gain (as determined by the high tension, HT, voltage applied to the photomultiplier tube, PMT, detector) can be accommodated. Lastly, the amount of stray light and dark current for the photomultiplier tube is determined. Though specific to the Jasco CD spectrometers characterized in this study, it is expected that all CD spectrometers exhibit similar behavior and the methodology described here can be usefully applied to characterize CD spectrometers independent of manufacturer.


Identification and phylogenetic analysis of toxigenic cyanobacteria by multiplex randomly amplified polymorphic DNA PCR.
Randomly amplified polymorphic DNA PCR was used to generate unique and identifying DNA profiles for members of the cyanobacterial genera Anabaena and Microcystis, which are responsible for much of the production of nuisance blooms in various freshwater systems, including recreational and drinking water supplies. A method based on the combination of two 10-mer oligonucleotides in a single PCR was developed to provide specific and repeatable DNA fingerprints for cyanobacterial isolates. The strain-specific randomly amplified polymorphic DNA profiles made it possible to discriminate among all toxigenic cyanobacteria studied to the three taxonomic levels of genus, species, and strain. Analysis of DNA typing results obtained by the described method clearly distinguishes between the genera Anabaena and Microcystis. The markers produced for each strain were also applied to a phylogenetic analysis to infer genetic relatedness in this group of prokaryotes.


Industrial membrane processes in the treatment of process waters and liquors.
A review on pulp and paper industrial membrane processes using a variety of modules and processes is presented. Membranes are mostly used today to purify process waters and to recover coating colours. Ultrafiltration using tubular membrane modules or cross-rotational (CR) filtration has been widely applied for the purification of process waters. The reuse of UF membrane permeate has decreased the fresh water consumption to lower than 6 m³/t of paper in some paper machines. Some industrial membrane processes also recover valuable products from different streams (e.g lignosulphonates). Membranes are also combined with biological degradation processes in some paper mills. Nanofiltration has been used to purify the effluents discharged from the activated sludge process. At least two reverse osmosis plants purify river water to be used as raw water in the mill. Furthermore, advantages of different membrane modules and the current ways to treat membrane concentrate are discussed.


Exploring global Cryptosporidium emissions to surface water.
The protozoan parasite Cryptosporidium is a major cause of diarrhoea worldwide. This paper presents the first model-based inventory with 0.5 by 0.5 degree resolution of global Cryptosporidium emissions for the year 2000 from humans and animals to surface water. The model is based on nutrient distribution modelling, because the sources and transport of oocysts and nutrients to the surface water are comparable. Total emissions consist of point source emissions from wastewater and nonpoint source emissions by runoff of oocysts in manure from agricultural lands. Results indicate a global emission of 3 × 10(17) oocysts per year, with comparable contributions from point and nonpoint sources. Hot-spot areas for point sources are big cities in China, India and Latin America, while the area with the largest nonpoint source emissions is in China. Uncertainties in the model are large. Main areas for further study are (i) excretion rates of oocysts by humans and animals, (ii) emissions of humans not connected to sewage systems, and (iii) retention of oocysts to determine surface water pathogen concentrations rather than emissions. Our results are useful to health organisations to identify priority areas for further study and intervention.


Lyme borreliosis in human patients in Florida and Georgia, USA.
The aim of this study was to determine the cause of illness in several human patients residing in Florida and Georgia, USA, with suspected Lyme disease based upon EM-like skin lesions and/or symptoms consistent with early localized or late disseminated Lyme borreliosis. Using polymerase chain reaction (PCR) assays developed specifically for Lyme group Borrelia spp., followed by DNA sequencing for confirmation, we identified Borrelia burgdorferi sensu lato DNA in samples of blood and skin and also in lone star ticks (Amblyomma americanum) removed from several patients who either live in or were exposed to ticks in Florida or Georgia. This is the first report to present combined PCR and DNA sequence evidence of infection with Lyme Borrelia spp. in human patients in the southern U.S., and to demonstrate that several B. burgdorferi sensu lato species may be associated with Lyme disease-like signs and symptoms in southern states. Based on the findings of this study, we suggest that human Lyme borreliosis occurs in Florida and Georgia, and that some cases of Lyme-like illness referred to as southern tick associated rash illness (STARI) in the southern U.S. may be attributable to previously undetected B. burgdorferi sensu lato infections.


The role of anti-müllerian hormone (AMH) in assessing ovarian reserve.
Anti-müllerian hormone (AMH) has been suggested as a marker for the quantity of oocytes remaining within the ovaries (ovarian reserve). It was shown to correlate with antral follicle counts (AFC), outcomes from ovarian stimulation, and onset of menopause. Thus, AMH was previously considered to be the ideal marker of ovarian reserve because it is exclusively produced by granulosa cells and is the only marker that was thought to be stable throughout the menstrual cycle. However, recent studies demonstrate fluctuations in AMH levels during the menstrual cycles, questioning the utility of AMH as a marker of oocyte quantity. We report the case of a 32-yr-old Gravida 0 woman with idiopathic hypogonadotropic hypogonadism who presented for fertility treatment with unstable AMH levels. The patient's initial FSH and LH levels were both below 1.0 mIU/ml. Estradiol was 28 pg/ml. Her initial AMH and AFC were 0.20 ng/ml and 0, respectively. She underwent three cycles of fertility treatment. During the 16-wk course of treatment with human menopausal gonadotropins, normal follicular development was observed. Both AMH and AFC gradually increased during treatment and peaked at 1.26 ng/ml and 6, respectively. On the third cycle of treatment, she successfully conceived. In the case of idiopathic hypogonadotropic hypogonadism, AMH concentration increases because human menopausal gonadotropin stimulates the growth of FSH-dependent follicles. Thus, AMH has limitations because it only reflects the growing follicular pool that is responsive to gonadotropins. Therefore, AMH may not be solely reflective of the underlying primordial pool.


Electron microscopy of growing oocytes of Rana pipiens.
1. In the cytoplasm of oocytes of stage Y(0), prior to the appearance of yolk, one observes a few scattered profiles of endoplasmic reticulum and numerous filamentous mitochondria, usually distributed at random but sometimes clustered. As the nuclear membrane begins to bulge outward, small granules and short rods appear in the perinuclear cytoplasm and endoplasmic reticulum becomes more prominent throughout the cytoplasm. 2. Coincident with the appearance of the first yolk platelets, which are deposited in a narrow peripheral ring within the endoplasm at stage Y(1), protoplasmic processes, the microvilli, push out all over the surface of the oocyte. At the same time follicle cells pull away but remain attached to the oocyte at some points through finger-like processes which interdigitate with neighboring microvilli. It is estimated that the microvilli increase the absorptive area of the surface to about thirty-five times that of a simple sphere. Just beneath the microvillous layer is the basal protoplasm of the cortex, now containing tiny granules probably synthesized from newly absorbed raw materials. Cortical granules appear and become aligned below the basal layer on the external border of the endoplasm. Both the cortical granules and the yolk platelets measure up to 1 micro in diameter at this stage. 3. By stage Y(3) (yolk filling peripheral three-fourths of cytoplasm), the basal layer of the cortex is folded so that it appears in section as alternating ridges and valleys. The microvilli now extend from the summits of the cortical ridges. Small, ring-shaped granules are abundant in the cortex. Cortical granules have increased to 2 micro in diameter. 4. Yolk platelets continue to be synthesized around the cortical granules and in the subjacent endoplasm. The largest platelets measured in the interior cytoplasm at stage Y(4) (cytoplasm filled with yolk) were 3.7 micro wide by 5.8 micro long. Pigment granules increase in size from 0.15 micro in diameter at stage Y(3) to 0.30 micro in diameter at stage Y(4).


[Borrelial lymphocytoma].
Borrelial lymphocytoma is a relatively rare but typical presentation of Lyme disease. Predilection sites are the ears in children and chest/nipples in adults. It is treated like an erythema migrans and has a good prognosis. A 16-year-old boy presented with a swollen, red and painful right nipple since several months. An ultrasound showed normal breast tissue. The patient was referred to the pediatric surgeon who performed an incision biopsy. Histopathological examination revealed follicular hyperplasia without signs of malignancy. An infectious cause, most likely Lyme disease, was suspected. Serological analysis and PCR of the tissue confirmed the diagnosis of a borrelial lymphocytoma, and the patient was treated with doxycycline with good result. Early recognition of the characteristic clinical presentation of borrelial lymphocytoma, supported by positive results from serologic testing for Lyme disease, avoids the need for additional and invasive diagnostic tests.


Low adherence to antidepressants is associated with increased mortality following stroke: A large nationally representative cohort study.
Post stroke depression is common and pervasive. In the general population, there has been some controversy that antidepressant (AD) medication is associated with premature mortality. Data is still lacking regarding the association between adherence to antidepressants (AD) and all-cause mortality. In this retrospective analysis of a population-based cohort of patients, 32,361 post-stroke patients who purchased at least one AD were followed for all-cause mortality over 4-years. Adherence to AD was measured as a ratio between dispensed and prescribed durations and was modeled as: non-adherence (<20%, n=8619), poor (20-50%, n=5108), moderate (50-80%, n=5656), and good (>80%, n=12,978) adherence. Multivariable survival analyses, adjusted for demographic and clinical variables including physical comorbidities known to influence mortality, were conducted. Unadjusted mortality rates were 16.5%, 20.2%, 22.2% and 23.7% in those classified as non-adherent, poor, moderate and good adherence respectively (χ2=174.6, p<0.0001). In the adjusted model, the non-adherent and poor adherence groups had significantly increased mortality Hazard Ratios (HR) of 1.25 (95% CI: 1.17-1.33) and 1.17 (95% CI: 1.09-1.26) respectively compared to the good adherence group. This nationally representative data suggests that poor adherence to AD is associated with increased all-cause mortality among people who had a stroke. Given our findings and the high prevalence of anxiety and depression along with AD effectiveness, efforts to promote AD adherence in this population may be warranted in clinical practice.


AAV CRISPR editing rescues cardiac and muscle function for 18 months in dystrophic mice.
Adeno-associated virus-mediated (AAV-mediated) CRISPR editing is a revolutionary approach for treating inherited diseases. Sustained, often life-long mutation correction is required for treating these diseases. Unfortunately, this has never been demonstrated with AAV CRISPR therapy. We addressed this question in the mdx model of Duchenne muscular dystrophy (DMD). DMD is caused by dystrophin gene mutation. Dystrophin deficiency leads to ambulation loss and cardiomyopathy. We treated 6-week-old mice intravenously and evaluated disease rescue at 18 months. Surprisingly, nominal dystrophin was restored in skeletal muscle. Cardiac dystrophin was restored, but histology and hemodynamics were not improved. To determine the underlying mechanism, we evaluated components of the CRISPR-editing machinery. Intriguingly, we found disproportional guide RNA (gRNA) vector depletion. To test whether this is responsible for the poor outcome, we increased the gRNA vector dose and repeated the study. This strategy significantly increased dystrophin restoration and reduced fibrosis in all striated muscles at 18 months. Importantly, skeletal muscle function and cardiac hemodynamics were significantly enhanced. Interestingly, we did not see selective depletion of the gRNA vector after intramuscular injection. Our results suggest that gRNA vector loss is a unique barrier for systemic AAV CRISPR therapy. This can be circumvented by vector dose optimization.


The international limits and population at risk of Plasmodium vivax transmission in 2009.
A research priority for Plasmodium vivax malaria is to improve our understanding of the spatial distribution of risk and its relationship with the burden of P. vivax disease in human populations. The aim of the research outlined in this article is to provide a contemporary evidence-based map of the global spatial extent of P. vivax malaria, together with estimates of the human population at risk (PAR) of any level of transmission in 2009. The most recent P. vivax case-reporting data that could be obtained for all malaria endemic countries were used to classify risk into three classes: malaria free, unstable (<0.1 case per 1,000 people per annum (p.a.)) and stable (> or =0.1 case per 1,000 p.a.) P. vivax malaria transmission. Risk areas were further constrained using temperature and aridity data based upon their relationship with parasite and vector bionomics. Medical intelligence was used to refine the spatial extent of risk in specific areas where transmission was reported to be absent (e.g., large urban areas and malaria-free islands). The PAR under each level of transmission was then derived by combining the categorical risk map with a high resolution population surface adjusted to 2009. The exclusion of large Duffy negative populations in Africa from the PAR totals was achieved using independent modelling of the gene frequency of this genetic trait. It was estimated that 2.85 billion people were exposed to some risk of P. vivax transmission in 2009, with 57.1% of them living in areas of unstable transmission. The vast majority (2.59 billion, 91.0%) were located in Central and South East (CSE) Asia, whilst the remainder were located in America (0.16 billion, 5.5%) and in the Africa+ region (0.10 billion, 3.5%). Despite evidence of ubiquitous risk of P. vivax infection in Africa, the very high prevalence of Duffy negativity throughout Central and West Africa reduced the PAR estimates substantially. After more than a century of development and control, P. vivax remains more widely distributed than P. falciparum and is a potential cause of morbidity and mortality amongst the 2.85 billion people living at risk of infection, the majority of whom are in the tropical belt of CSE Asia. The probability of infection is reduced massively across Africa by the frequency of the Duffy negative trait, but transmission does occur on the continent and is a concern for Duffy positive locals and travellers. The final map provides the spatial limits on which the endemicity of P. vivax transmission can be mapped to support future cartographic-based burden estimations.


SLURP-1 modulates corneal homeostasis by serving as a soluble scavenger of urokinase-type plasminogen activator.
Our previous study revealed the immunomodulatory property of the secreted lymphocyte antigen (Ly6)/urokinase-type plasminogen activator receptor (uPAR)-related protein-1 (SLURP1), abundantly expressed in the cornea and associated with the hyperkeratotic disorder Mal de Meleda. Here, we test the hypothesis that SLURP1 modulates the functions of membrane-tethered uPAR by acting as a soluble scavenger of its ligand urokinase-type plasminogen activator (uPA). Human corneal limbal epithelial (HCLE) and mouse corneal stromal fibroblast MK/T-1 cells were employed to examine the effect of SLURP1 on cell proliferation and migration. Human corneal limbal epithelial cell clones stably expressing SLURP1 under the control of cytomegalovirus (CMV) promoter were generated using lentiviral vectors. Recombinant 6× His-mouse Slurp1 and maltose-binding protein (MBP)-mouse uPA were expressed in Escherichia coli and partially purified using nickel-ion and amylose columns, respectively. Slurp1 interaction with uPA was detected using ligand blots, ELISA, pull-down assays, and immunofluorescent staining. Stable expression of SLURP1 in HCLE cells was confirmed by immunoblots and immunofluorescent staining. Human corneal limbal epithelial and MK/T-1 cell proliferation and migration rates were suppressed by exogenous SLURP1. Ligand blots, ELISA, and pull-down assays indicated that Slurp1 efficiently interacts with uPA. Immunofluorescent staining demonstrated that exogenous SLURP1 decreased the amount of cell surface-bound uPA in the leading edges of migrating cells. In gap-filling assays, wild-type HCLE cells responded to uPA by increasing their velocity and closing larger area, while the SLURP1-expressing HCLE cells failed to do so. SLURP1 modulates corneal homeostasis by serving as a soluble scavenger of uPA and regulating the uPA-dependent functions of uPAR.


Localization of pigment epithelium derived factor (PEDF) in developing and adult human ocular tissues.
To localize pigment epithelium-derived factor (PEDF) in developing and adult human ocular tissues. PEDF was localized in fetal and adult eyes by immunofluorescence with a polyclonal antibody (pAb) against amino acids 327-343 of PEDF, or a monoclonal antibody (mAb) against the C-terminal 155 amino acids of PEDF. Specificity of the antibodies was documented by Western blotting. PEDF mRNA was localized in adult retina by in situ hybridization. In developing retinas (7.4 to 21.5 fetal weeks, Fwks), pAb anti-PEDF labeled retinal pigment epithelium (RPE) granules, developing cones, some neuroblasts and many cells in the ganglion cell layer (GCL). In adult retinas, pAb anti-PEDF labeled rod and cone cytoplasm and nuclei of rods but not cones. Cells in the INL and GCL, choroid, corneal epithelium and endothelium, and ciliary body were also pAb PEDF-positive. Preadsorption of pAb anti-PEDF with the immunizing peptide blocked specific labeling in retina and other tissues, except for photoreceptor outer segments. In agreement with the immunolocalization with pAb anti-PEDF, in situ hybridization revealed PEDF mRNA in the RPE, photoreceptors, inner nuclear layer cells and ganglion cells in adult retina. In developing retinas 18 Fwks and older, and in adult retinas, mAb anti-PEDF labeled the interphotoreceptor matrix (IPM). Western blots of retina, cornea, and ciliary body/iris with pAb anti-PEDF produced several bands at about 46 kDa. With mAb anti-PEDF, retina produced one band at about 46 kDa; cornea and ciliary body/iris had several bands at about 46 kDa. PEDF, originally reported as a product of RPE cells, is present in photoreceptors and inner retinal cell types in developing and adult human eyes. Photoreceptors and RPE may secrete PEDF into the IPM.


Megakaryopoiesis and platelet production: insight into hematopoietic stem cell proliferation and differentiation.
Hematopoietic stem cells (HSCs) undergo successive lineage commitment steps to generate megakaryocytes (MKs) in a process referred to as megakaryopoiesis. MKs undergo a unique differentiation process involving endomitosis to eventually produce platelets. Many transcription factors participate in the regulation of this complex progress. Chemokines and other factors in the microenvironment where megakaryopoiesis and platelet production occur play vital roles in the regulation of HSC lineage commitment and MK maturation; among these factors, thrombopoietin (TPO) is the most important. Endomitosis is a vital process of MK maturation, and granules that are formed in MKs are important for platelet function. Proplatelets are firstly generated from mature MKs and then become platelets. The proplatelet production process was verified by novel studies that revealed that the mechanism is partially regulated by the invaginated membrane system (IMS), microtubules and Rho GTPases. The extracellular matrices (ECMs) and shear stress also affect and regulate the process while the mature MKs migrate from the marrow to the sub-endothelium region near the venous sinusoids leading to the release of platelets into the circulation. This review describes the entire process of megakaryopoiesis in detail, illustrates both the transcriptional and microenvironmental regulation of MKs and provides insight into platelet biogenesis.


The bacteriological quality of drinking water in Haldwani block of Nainital district, Uttarakhand, India.
A cross-sectional study was conducted to assess the bacteriological quality of water in Haldwani block, Nainital District, India. Stratified random sampling was used to categorize water sources and consumer points. In total, 108 samples were collected: 15 from the Gola river, 51 from water taps, 24 from water treatment plants and 18 from tube wells. Samples were tested for coliforms by the most probable number technique. Identification of species was done by standard procedures. Of 108 water samples, 58.8% were found to be polluted. All samples of water (n = 15) from different sites of the Gola river were found to be highly contaminated. Out of 24 water treatment plant samples, four samples were found unsatisfactory, while more than half (51.6%) of its supplies to water taps were polluted. From tube wells and their water taps, 88.8 and 60% samples were found safe for drinking respectively. Bacterial contamination of water treatment plants and their supplies indicates significant disparities in the efficiency of water treatment processes. Contamination of water taps of tube wells suggests leakage of pipes. There is an urgent need to improve these services to ensure the supply of safe water for consumers.


Information filtering via a scaling-based function.
Finding a universal description of the algorithm optimization is one of the key challenges in personalized recommendation. In this article, for the first time, we introduce a scaling-based algorithm (SCL) independent of recommendation list length based on a hybrid algorithm of heat conduction and mass diffusion, by finding out the scaling function for the tunable parameter and object average degree. The optimal value of the tunable parameter can be abstracted from the scaling function, which is heterogeneous for the individual object. Experimental results obtained from three real datasets, Netflix, MovieLens and RYM, show that the SCL is highly accurate in recommendation. More importantly, compared with a number of excellent algorithms, including the mass diffusion method, the original hybrid method, and even an improved version of the hybrid method, the SCL algorithm remarkably promotes the personalized recommendation in three other aspects: solving the accuracy-diversity dilemma, presenting a high novelty, and solving the key challenge of cold start problem.


Abortion and mental health: findings from The National Comorbidity Survey-Replication.
To examine whether a first abortion increases risk of mental health disorders compared with a first childbirth with and without considering prepregnancy mental health and adverse exposures, childhood economic status, miscarriage history, age at first abortion or childbirth, and race or ethnicity. A cohort study compared rates of mental disorders (anxiety, mood, impulse-control, substance use, eating disorders, and suicidal ideation) among 259 women postabortion and 677 women postchildbirth aged 18-42 years at the time of interview from The National Comorbidity Survey-Replication. The percentage of women with no, one, two, and three or more mental health disorders before their first abortion was 37.8%, 19.7%, 15.2%, and 27.3% and before their first childbirth was 57.9%, 19.6%, 9.2%, and 13.3%, respectively, indicating that women in the abortion group had more prior mental health disorders than women in the childbirth group (P<.001). Although in unadjusted Cox proportional hazard models, abortion compared with childbirth was associated with statistically significant higher hazards of postpregnancy mental health disorders, associations were reduced and became nonstatistically significant for five disorders after adjusting for the aforementioned factors. Hazard ratios and associated 95% confidence intervals dropped from 1.52 (1.08-2.15) to 1.12 (0.87-1.46) for anxiety disorders; from 1.56 (1.23-1.98) to 1.18 (0.88-1.56) for mood disorders; from 1.62 (1.02-2.57) to 1.10 (0.75-1.62) for impulse-control disorders; from 2.53 (1.09-5.86) to 1.82 (0.63-5.25) for eating disorders; and from 1.62 (1.09-2.40) to 1.25 (0.88-1.78) for suicidal ideation. Only the relationship between abortion and substance use disorders remained statistically significant, although the hazard ratio dropped from 3.05 (1.94-4.79) to 2.30 (1.35-3.92). After accounting for confounding factors, abortion was not a statistically significant predictor of subsequent anxiety, mood, impulse-control, and eating disorders or suicidal ideation. LEVEL OF EVEDIENCE: II.


The acute effects of low-dose TNF-α on glucose metabolism and β-cell function in humans.
Type 2 diabetes is characterized by increased insulin resistance and impaired insulin secretion. Type 2 diabetes is also associated with low-grade inflammation and increased levels of proinflammatory cytokines such as TNF-α. TNF-α has been shown to impair peripheral insulin signaling in vitro and in vivo. However, it is unclear whether TNF-α may also affect endogenous glucose production (EGP) during fasting and glucose-stimulated insulin secretion (GSIS) in vivo. We hypothesized that low-dose TNF- α would increase EGP and attenuate GSIS. Recombinant human TNF-α or placebo was infused in healthy, nondiabetic young men (n = 10) during a 4-hour basal period followed by an intravenous glucose tolerance test (IVGTT). TNF-α lowered insulin levels by 12% during the basal period (P < 0.05). In response to the IVGTT, insulin levels increased markedly in both trials, but there was no difference between trials. Compared to placebo, TNF-α did not affect EGP during the basal period. Our results indicate that TNF-α acutely lowers basal plasma insulin levels but does not impair GSIS. The mechanisms behind this are unknown but we suggest that it may be due to TNF-α increasing clearance of insulin from plasma without impairing beta-cell function or hepatic insulin sensitivity.


A full genome screen for autism with evidence for linkage to a region on chromosome 7q. International Molecular Genetic Study of Autism Consortium.
Autism is characterized by impairments in reciprocal social interaction and communication, and restricted and sterotyped patterns of interests and activities. Developmental difficulties are apparent before 3 years of age and there is evidence for strong genetic influences most likely involving more than one susceptibility gene. A two-stage genome search for susceptibility loci in autism was performed on 87 affected sib pairs plus 12 non-sib affected relative-pairs, from a total of 99 families identified by an international consortium. Regions on six chromosomes (4, 7, 10, 16, 19 and 22) were identified which generated a multipoint maximum lod score (MLS) > 1. A region on chromosome 7q was the most significant with an MLS of 3.55 near markers D7S530 and D7S684 in the subset of 56 UK affected sib-pair families, and an MLS of 2.53 in all 87 affected sib-pair families. An area on chromosome 16p near the telomere was the next most significant, with an MLS of 1.97 in the UK families, and 1.51 in all families. These results are an important step towards identifying genes predisposing to autism; establishing their general applicability requires further study.


Vitamin D and infectious diseases.
In the early 1920s the antirachitic effect of food irradiated with ultraviolet light and cod liver oil has been recognized. The antirachitic substance was identified and called "vitamin D". Since then the key role of vitamin D in calcium and bone homeostasis has been investigated. Moreover, it has been recognized that vitamin D is able to modulate a variety of processes and regulatory systems such as host defense, inflammation, immunity, and repair. According to recent studies, vitamin D deficiency is likely to be an important etiological factor in the pathogenesis of many chronic diseases, as well as it has been associated with higher mortality rate for respiratory disease. In this regard, either observational studies aimed to verify an association between low vitamin D level and the incidence of respiratory tract infections (RTIs) or clinical trials on the effect of vitamin D as a supplementary treatment in RTIs patients have been presented in the emerging clinical literature. Conflicting results have been demonstrated in several randomized, double-blind, placebo controlled trials concerning the vitamin D treatment in tuberculosis. Some studies suggest a beneficial effect by vitamin D but it could not be reproduced in larger studies so far. In conclusion, although basic science research suggests that vitamin D may play an important role in modulating immune functions, no strong evidence exists whether correction of vitamin D depletion may be useful in the prevention or treatment of infections. Further and larger studies may clarify the role of vitamin D in infection.


Campylobacter-Acanthamoeba interactions.
Campylobacter jejuni is a foodborne pathogen recognized as the major cause of human bacterial enteritis. Undercooked poultry products and contaminated water are considered as the most important sources of infection. Some studies suggest transmission and survival of this bacterial pathogen may be assisted by the free-living protozoa Acanthamoeba. The latter is known to play the role of a host for various pathogenic bacteria, protecting them from harsh environmental conditions. Importantly, there is a similarity between the mechanisms of bacterial survival within amoebae and macrophages, making the former a convenient tool for the investigation of the survival of pathogenic bacteria in the environment. However, the molecular mechanisms involved in the interaction between Campylobacter and Acanthamoeba are not well understood. Whilst some studies suggest the ability of C. jejuni to survive within the protozoa, the other reports support an extracellular mode of survival only. In this review, we focus on the studies investigating the interaction between Campylobacter and Acanthamoeba, address some reasons for the contradictory results, and discuss possible implications of these results for epidemiology. Additionally, as the molecular mechanisms involved remain unknown, we also suggest possible factors that may be involved in this process. Deciphering the molecular mechanisms of pathogen-protozoa interaction will assist in a better understanding of Campylobacter lifestyle and in the development of novel antibacterial drugs.


Sigma: multiple alignment of weakly-conserved non-coding DNA sequence.
Existing tools for multiple-sequence alignment focus on aligning protein sequence or protein-coding DNA sequence, and are often based on extensions to Needleman-Wunsch-like pairwise alignment methods. We introduce a new tool, Sigma, with a new algorithm and scoring scheme designed specifically for non-coding DNA sequence. This problem acquires importance with the increasing number of published sequences of closely-related species. In particular, studies of gene regulation seek to take advantage of comparative genomics, and recent algorithms for finding regulatory sites in phylogenetically-related intergenic sequence require alignment as a preprocessing step. Much can also be learned about evolution from intergenic DNA, which tends to evolve faster than coding DNA. Sigma uses a strategy of seeking the best possible gapless local alignments (a strategy earlier used by DiAlign), at each step making the best possible alignment consistent with existing alignments, and scores the significance of the alignment based on the lengths of the aligned fragments and a background model which may be supplied or estimated from an auxiliary file of intergenic DNA. Comparative tests of sigma with five earlier algorithms on synthetic data generated to mimic real data show excellent performance, with Sigma balancing high "sensitivity" (more bases aligned) with effective filtering of "incorrect" alignments. With real data, while "correctness" can't be directly quantified for the alignment, running the PhyloGibbs motif finder on pre-aligned sequence suggests that Sigma's alignments are superior. By taking into account the peculiarities of non-coding DNA, Sigma fills a gap in the toolbox of bioinformatics.


Rates of and Factors Associated With Placebo Response in Trials of Pharmacotherapies for Nonalcoholic Steatohepatitis: Systematic Review and Meta-analysis.
It is important to know the extent of the placebo effect in designing randomized controlled trials for patients with nonalcoholic steatohepatitis (NASH), to accurately calculate sample size and define treatment endpoints. We performed a systematic review and meta-analysis of the placebo groups from randomized controlled trials of adults with NASH that provided histologic and/or magnetic resonance image-based assessments. We identified trials through a comprehensive search of MEDLINE, EMBASE, Cochrane Central Register of Controlled Trials, and Scopus, from each database's inception through January 2, 2018. We identified 39 randomized controlled trials, comprising 1463 patients who received placebo. Histologic assessment data (the nonalcoholic fatty liver disease activity scores, NAS) were available from 956 patients; magnetic resonance spectroscopy data were available from 295 patients and magnetic resonance proton density fat fraction measurements from 61 patients. Overall, 25% of patients given placebo had an improvement in NAS by 2 or more points (95% CI, 21%-29%) with a small amount of heterogeneity (I2 = 27%). There were improvements by at least 1 point in steatosis scores of 33% ± 3% of patients, in hepatocyte ballooning scores of 30% ± 3% of patients, in lobular inflammation scores of 32% ± 3% of patients, and in fibrosis scores of 21% ± 3% of patients, with a moderate amount of heterogeneity among trials (I2 range, 51%-63%). Patients given placebo had a statistically significant improvement in NAS (by 0.72 ± 0.19), with a large amount of heterogeneity (I2 = 96%). Univariate and multivariate meta-regression showed that trials with a higher baseline NAS, those conducted in South America, and those in which patients had a decrease in body mass index, were associated with greater improvements in NAS among patients given placebo. Patients given placebo had significant reductions in intrahepatic triglyceride, measured by magnetic resonance spectroscopy (by 1.45% ± 0.54%) with moderate heterogeneity (I2 = 40%), and in magnetic resonance proton density fat fraction (by 2.43 ± 0.89), without heterogeneity (I2 = 0). Mean serum levels of alanine and aspartate aminotransferases decreased significantly (by 11.7 ± 3.8 U/L and 5.9 ± 2.1 U/L, respectively; P < .01 for both). In a meta-analysis of randomized controlled trials of NASH, patients given placebo have significant histologic, radiologic, and biochemical responses. The placebo response should be considered in designing trials of agents for treatment of NASH.


Imaging of the Yellow Cameleon 3.6 indicator reveals that elevations in cytosolic Ca2+ follow oscillating increases in growth in root hairs of Arabidopsis.
In tip-growing cells, the tip-high Ca(2+) gradient is thought to regulate the activity of components of the growth machinery, including the cytoskeleton, Ca(2+)-dependent regulatory proteins, and the secretory apparatus. In pollen tubes, both the Ca(2+) gradient and cell elongation show oscillatory behavior, reinforcing the link between the two. We report that in growing root hairs of Arabidopsis (Arabidopsis thaliana), an oscillating tip-focused Ca(2+) gradient can be resolved through imaging of a cytosolically expressed Yellow Cameleon 3.6 fluorescence resonance energy transfer-based Ca(2+) sensor. Both elongation of the root hairs and the associated tip-focused Ca(2+) gradient show a similar dynamic character, oscillating with a frequency of 2 to 4 min(-1). Cross-correlation analysis indicates that the Ca(2+) oscillations lag the growth oscillations by 5.3 +/- 0.3 s. However, growth never completely stops, even during the slow cycle of an oscillation, and the concomitant tip Ca(2+) level is always slightly elevated compared with the resting Ca(2+) concentration along the distal shaft, behind the growing tip. Artificially increasing Ca(2+) using the Ca(2+) ionophore A23187 leads to immediate cessation of elongation and thickening of the apical cell wall. In contrast, dissipating the Ca(2+) gradient using either the Ca(2+) channel blocker La(3+) or the Ca(2+) chelator EGTA is accompanied by an increase in the rate of cell expansion and eventual bursting of the root hair tip. These observations are consistent with a model in which the maximal oscillatory increase in cytosolic Ca(2+) is triggered by cell expansion associated with tip growth and plays a role in the subsequent restriction of growth.


Pupillometry to differentiate idiopathic hypersomnia from narcolepsy type 1.
Idiopathic hypersomnia is poorly diagnosed in the absence of biomarkers to distinguish it from other central hypersomnia subtypes. Given that light plays a main role in the regulation of sleep and wake, we explored the retinal melanopsin-based pupil response in patients with idiopathic hypersomnia and narcolepsy type 1, and healthy subjects. Twenty-seven patients with narcolepsy type 1 (women 59%, 36 ± 11.5 years old), 36 patients with idiopathic hypersomnia (women 83%, 27.2 ± 7.2 years old) with long total sleep time (> 11/24 hr), and 43 controls (women 58%, 30.6 ± 9.3 years old) were included in this study. All underwent a pupillometry protocol to assess pupil diameter, and the relative post-illumination pupil response to assess melanopsin-driven pupil responses in the light non-visual input pathway. Differences between groups were assessed using logistic regressions adjusted on age and sex. We found that patients with narcolepsy type 1 had a smaller baseline pupil diameter as compared with idiopathic hypersomnia and controls (p < 0.05). In addition, both narcolepsy type 1 and idiopathic hypersomnia groups had a smaller relative post-illumination pupil response (respectively, 31.6 ± 13.9% and 33.2 ± 9.9%) as compared with controls (38.7 ± 9.7%), suggesting a reduced melanopsin-mediated pupil response in both types of central hypersomnia (p < 0.01). Both narcolepsy type 1 and idiopathic hypersomnia showed a smaller melanopsin-mediated pupil response, and narcolepsy type 1, unlike idiopathic hypersomnia, also displayed a smaller basal pupil diameter. Importantly, we found that the basal pupil size permitted to well discriminate idiopathic hypersomnia from narcolepsy type 1 with a specificity = 66.67% and a sensitivity = 72.22%. Pupillometry may aid to multi-feature differentiation of central hypersomnia subtypes.


Clinical implications of NSAID pharmacokinetics: special populations, special considerations.
Nonsteroidal anti-inflammatory drugs (NSAIDs) are the centerpiece of pharmacologic therapy for most rheumatic disorders and related conditions, and as such are used in great numbers. These drugs are relatively safe and effective, but the pharmacokinetics of NSAIDs can be substantially altered in certain groups of patients, including the elderly and patients with renal and hepatic disease. In these patients, the risk of NSAID toxicity is increased. An understanding of NSAID pharmacokinetics in these groups can help physicians to adjust therapeutic regimens in order to limit the potentially serious complications of long-term NSAID therapy. The author discusses the age-related physiologic changes that may affect the various areas of drug pharmacokinetics and ways in which NSAIDs may interact with other drugs. Risk assessment and monitoring methods are also discussed.


Macrophage subset sensitivity to endotoxin tolerisation by Porphyromonas gingivalis.
Macrophages (MΦs) determine oral mucosal responses; mediating tolerance to commensal microbes and food whilst maintaining the capacity to activate immune defences to pathogens. MΦ responses are determined by both differentiation and activation stimuli, giving rise to two distinct subsets; pro-inflammatory M1- and anti-inflammatory/regulatory M2- MΦs. M2-like subsets predominate tolerance induction whereas M1 MΦs predominate in inflammatory pathologies, mediating destructive inflammatory mechanisms, such as those in chronic P.gingivalis (PG) periodontal infection. MΦ responses can be suppressed to benefit either the host or the pathogen. Chronic stimulation by bacterial pathogen associated molecular patterns (PAMPs), such as LPS, is well established to induce tolerance. The aim of this study was to investigate the susceptibility of MΦ subsets to suppression by P. gingivalis. CD14(hi) and CD14(lo) M1- and M2-like MΦs were generated in vitro from the THP-1 monocyte cell line by differentiation with PMA and vitamin D3, respectively. MΦ subsets were pre-treated with heat-killed PG (HKPG) and PG-LPS prior to stimulation by bacterial PAMPs. Modulation of inflammation was measured by TNFα, IL-1β, IL-6, IL-10 ELISA and NFκB activation by reporter gene assay. HKPG and PG-LPS differentially suppress PAMP-induced TNFα, IL-6 and IL-10 but fail to suppress IL-1β expression in M1 and M2 MΦs. In addition, P.gingivalis suppressed NFκB activation in CD14(lo) and CD14(hi) M2 regulatory MΦs and CD14(lo) M1 MΦs whereas CD14(hi) M1 pro-inflammatory MΦs were refractory to suppression. In conclusion, P.gingivalis selectively tolerises regulatory M2 MΦs with little effect on pro-inflammatory CD14(hi) M1 MΦs; differential suppression facilitating immunopathology at the expense of immunity.


Nitric oxide mediates seasonal muscle potentiation in clam gills.
The physiology and timing of gill muscle potentiation were explored in the clam Mercenaria mercenaria. When isolated demibranchs were exposed twice (with an intervening wash) to the same concentration of 5-hydroxytryptamine, the second contraction was larger than the first. This potentiation was seasonal: it was present from November through June, and absent from July through October. Potentiation was not affected by the geographic origin of the clams, nor by their acclimation temperature. Potentiation was inhibited by the nitric oxide synthase (NOS) inhibitor L-NAME and mimicked by the nitric oxide (NO) donor DEANO. During the season of potentiation, immunoreactive NOS appeared in the gill muscles and the gill filament epithelium, but during the off-season, the enzyme occurred at the base of the gill filaments. Potentiation was inhibited by ODQ, which inhibits soluble guanylate cyclase (sGC), and it was mimicked by dibutyryl-cGMP, an analog of cyclic GMP (cGMP). Moreover, potentiation was inhibited by the protein kinase G (PKG) inhibitor Rp-8-CPT-cGMPS. During the season of potentiation, immunoreactive sGC was concentrated in the gill muscles and the gill filament epithelium; but during the off-season, immunoreactive sGC was found in the gill filament epithelium. These data suggest that the potentiation of gill muscle is mediated by a NO/cGMP/PKG signaling pathway.