Lifetime memories from persistently supple synapses.
It is here proposed that the evanescent network derived from malleable or supple synapses is the substrate for long-lasting memory. The subjective sense of memory permanence is not derived, as suggested by Bain and others, from the stabilization of synaptic structure which gives rise to consolidated distributed networks. This generally held wisdom that synapses are activated and ultimately stabilized to reflect the long-lasting substrate of memory is reinforced by increased interest in the importance of sparse coding in memory consolidation. One line of evidence for sparse coding derives from studies on the lateral nucleus neurons of the amygdala (for review, see Josselyn, 2010). These findings lead to the conclusion that a small number of neurons are both necessary and sufficient to retrieve the fear engram. Recently, it has been shown that sparse coding in the dentate gyrus of the hippocampus is sufficient for retrieving the fear conditioning engram (Liu et al., 2012). One implication of these findings is that memory is stored in selected cells and that this restricted storage is more or less permanent, as the authors note: "… Defined cell populations can form the cellular basis for fear memory engrams (Liu et al., 2012, p. 89)." But the problem with this model is that because new learning is incorporated into existing networks, stabilization would work against this integration. For this reason and because of obligatory processes of metabolic protein turnover and ongoing synaptic malleability, the "supple synapse" model is proposed (Routtenberg, A., Rekart, 2005; Routtenberg, 2008a,b). Specifically, long-lasting memory is derived from the instructive, use-dependent sampling of neural networks selected from a very large universe of networks that are evanescent because they are linked together by supple synapses. Importantly, if such suppleness did not exist, the stabilization of synapses would then prevent the physiological malleability of brain circuitry that is essential both for proper integrated information storage and for flexible information retrieval. A corollary of this proposal is that it suggests an alternative view of consolidation: the same agent which is disruptive immediately after learning is no longer effective later because the network, over time, becomes widely distributed and evanescent. Thus, time-dependence is replaced with space-dependence.


Follicular fluid C3a-peptide promotes oocyte maturation through F-actin aggregation.
Immature cumulus-oocyte complexes are retrieved to obtain mature oocytes by in vitro maturation (IVM), a laboratory tool in reproductive medicine to obtain mature oocytes. Unfortunately, the efficiency of IVM is not satisfactory. To circumvent this problem, we therefore intended to commence with the composition of ovarian follicular fluid (FF), an important microenvironment influencing oocyte growth. It is well known that FF has a critical role in oocyte development and maturation. However, the components in human FF remain largely unknown, particularly with regard to small molecular peptides. In current study, the follicular fluid derived from human mature and immature follicles were harvested. The peptide profiles of FF were further investigated by using combined ultrafiltration and LC-MS/MS. The differential peptides were preliminary determined by performing differentially expressed analysis. Human and mouse oocyte culture were used to verify the influence of differential peptides on oocyte development. Constructing plasmids, cell transfecting, Co-IP, PLA etc. were used to reveal the detail molecular mechanism. The results from differentially expressed peptide as well as cultured human and mouse oocytes analyses showed that highly conserved C3a-peptide, a cleavage product of complement C3a, definitely affected oocytes development. Intriguingly, C3a-peptide possessed a novel function that promoted F-actin aggregation and spindle migration, raised the percentage of oocytes at the MII stage, without increasing the chromosome aneuploidy ratio, especially in poor-quality oocytes. These effects of C3a-peptide were attenuated by C3aR morpholino inhibition, suggesting that C3a-peptide affected oocytes development by collaborating with its classical receptor, C3aR. Specially, we found that C3aR co-localized to the spindle with β-tubulin to recruit F-actin toward the spindle and subcortical region of the oocytes through specific binding to MYO10, a key regulator for actin organization, spindle morphogenesis and positioning in oocytes. Our results provide a new perspective for improving IVM culture systems by applying FF components and also provide molecular insights into the physiological function of C3a-peptide, its interaction with C3aR, and their roles in enabling meiotic division of oocytes.


Piperlongumine inhibits proliferation and survival of Burkitt lymphoma in vitro.
Piperlongumine (PL), a pepper plant alkaloid from Piper longum, kills solid tumor cells in a highly selective, potent fashion. To evaluate whether PL may have similar effects on malignant blood cells, we determined the efficacy with which PL inhibits the B-lymphocyte derived neoplasm, Burkitt lymphoma (BL). Low micromolar concentrations of PL (IC(50) = 2.8 μM × 8.5 μM) curbed growth and survival of two EBV(+) BL cell lines (Daudi, Raji) and two EBV BL cell lines (Ramos, DG-75), but left normal peripheral blood B-lymphocytes unharmed. PL-dependent cytotoxicity was effected in part by reduced NF-κB and MYC activity, with the former being caused by inhibition of IκBα degradation, nuclear translocation of p65, and binding of NF-κB dimers to cognate DNA sequences in gene promoters. In 4 of 4 BL cell lines, the NF-κB/MYC-regulated cellular target genes, E2F1 and MYB, were down regulated, while the stress sensor gene, GADD45B, was up regulated. The EBV-encoded oncogene, LMP-1, was suppressed in Daudi and Raji cells. Considering that NF-κB, MYC and LMP-1 play a crucial role in the biology of many blood cancers including BL, our results provide a strong preclinical rationale for considering PL in new intervention approaches for patients with hematologic malignancies.


Is the nexus necessary for cell-to-cell coupling of smooth muscle?
Electronmicroscopic study of electrically coupled smooth muscles was undertaken to determine the distribution of nexuses in various types of smooth muscle. The study revealed that while nexal structures were commonplace in some types of smooth muscle, they were very rare or absent in others, even though in some cases these cells were only a few nanometers distant from one another. The persistence in thin section of these structures in the main circular muscle of dog intestine after poor fixation, fixation under strain, cell shrinkage, and metabolic damage of various sorts seems to rule out the thesis that they are labile. The absence of nexuses in longitudinal muscle of dog intestine examined both by thin section and by freeze fracture suggests that in this tissue they are absent or very rare in vivo and cannot account for electrical coupling. Nexuses were discernible in thin sections of main circular muscle after a variety of experimental conditions of fixation. Metabolic inhibition or in vitro permanganate fixation partially destroyed nexal contacts. These procedures induced tissue, membrane apposition and an accompanying increase in the number of structures which resemble nexuses at low magnification (nexus-like structures). "Nexus-like" structures occurred in all smooth muscle fixed by in vitro permanganate associated with apposition of membranes and poor preservation of basement membrane. A technique of in vitro permanganate fixation was developed which prevented tissue swelling; consequently "nexus-like" structures were absent in tissues so treated. The suggestion is made that some structures described in the literature as nexuses, following permanganate fixation, may represent "nexus-like" structures. The balance of evidence suggests that nexuses need not be present for electrical coupling of some smooth muscle cells, in which other types of cell-to-cell contacts must be invoked.


Synthesis of biologically active N- and O-linked glycans with multisialylated poly-N-acetyllactosamine extensions using P. damsela α2-6 sialyltransferase.
Sialosides on N- and O-linked glycoproteins play a fundamental role in many biological processes, and synthetic glycan probes have proven to be valuable tools for elucidating these functions. Though sialic acids are typically found α2-3- or α2-6-linked to a terminal nonreducing end galactose, poly-LacNAc extended core-3 O-linked glycans isolated from rat salivary glands and human colonic mucins have been reported to contain multiple internal Neu5Acα2-6Gal epitopes. Here, we have developed an efficient approach for the synthesis of a library of N- and O-linked glycans with multisialylated poly-LacNAc extensions, including naturally occurring multisialylated core-3 O-linked glycans. We have found that a recombinant α2-6 sialyltransferase from Photobacterium damsela (Pd2,6ST) exhibits unique regioselectivity and is able to sialylate internal galactose residues in poly-LacNAc extended glycans which was confirmed by MS/MS analysis. Using a glycan microarray displaying this library, we found that Neu5Acα2-6Gal specific influenza virus hemagglutinins, siglecs, and plant lectins are largely unaffected by adjacent internal sialylation, and in several cases the internal sialic acids are recognized as ligands. Polyclonal IgY antibodies specific for internal sialoside epitopes were elicited in inoculated chickens.


Synovial sarcoma: a multivariate analysis of prognostic factors in 112 patients with primary localized tumors of the extremity.
Synovial sarcoma is a high-grade tumor that is associated with poor prognosis. Previous studies analyzing prognostic factors are limited because of inclusion of heterogeneous cohorts of patients with nonextremity and recurrent tumors. The objective of this study was to determine independent prognostic factors of primary synovial sarcoma localized to the extremity. Between July 1, 1982, and June 30, 1996, 112 patients underwent surgical resection for cure at our institution and then were followed-up prospectively. Clinical and pathologic factors examined for prognostic value included age, sex, tumor site and location, depth, size, microscopic status of surgical margins, invasion of bone or neurovascular structures, and monophasic or biphasic histology. The end points analyzed were the time to first local recurrence that was not preceded by a distant recurrence, time to any distant recurrence, and time to disease-related mortality. These end points were modeled using the method of Kaplan and Meier and analyzed by the log-rank test and Cox regression. The median duration of follow-up among survivors in this cohort of 112 patients was 72 months. The 5-year local-recurrence, distant-recurrence, and mortality rates were 12%, 39%, and 25%, respectively. Tumor size > or = 5 cm (P =.001; relative risk [RR] = 2. 7; 95% confidence interval [CI], 1.5 to 5.2) and the presence of bone or neurovascular invasion (P =.04; RR = 2.3; 95% CI, 1.0 to 5. 3) were independent adverse predictors of distant recurrence. Tumor size > or= 5 cm (P =.003; RR = 2.3; 95% CI, 1.4 to 6.3) and the presence of bone or neurovascular invasion (P =.03; RR = 2.7; 95% CI, 1.0 to 6.5) were also independent adverse predictors of mortality. The natural history of primary synovial sarcoma of the extremity is related to tumor size and invasion of bone and neurovascular structures.


Predictors of Response and Remission to Antidepressants in Geriatric Depression: A Systematic Review.
Geriatric depression is a heterogeneous disorder that increases morbidity and mortality in a population that is already vulnerable. Predicting response and remission to antidepressants could help clinicians to optimize the management of antidepressants and reduce the consequences of depression. The aim of this article is to present results of a systematic review of the literature on predictive factors related to antidepressant response and remission in older adults with depression. We identified sociodemographic, clinical, neuropsychological, neuroimaging, and genetic factors that could be potential predictors of outcomes. Inconsistent findings and methodological differences among studies, however, limit the generalizability and application of these predictors in clinical practice. The results of our review confirm that geriatric depression includes many subgroups of patients with particular endophenotypes that may influence the course of depression. Further studies are needed to characterize depression subgroups in order to better understand the pathophysiology of late life depression and to find specific predictors for each group of patients.


The corticothalamic system in sleep.
The transition from wakefulness to NREM sleep is associated with typical signs of brain electrical activity, characterized by prolonged periods of hyperpolarization and increased membrane conductance in thalamocortical (TC) neurons, with the consequence that incoming messages are inhibited and the cerebral cortex is deprived of signals from the outside world. There are three major oscillations during NREM sleep. Spindles are generated within the thalamus, due to thalamic reticular (RE) neurons that impose rhythmic inhibitory sequences onto TC neurons, but the widespread synchronization of this rhythm is governed by corticothalamic projections. There are two types of delta activity: clock-like waves generated in TC neurons by the interplay between two hyperpolarization-activated inward currents; and cortical waves that survive extensive thalamectomy. The hallmark of NREM sleep activity is the slow oscillation, generated intracortically, which has the virtue of grouping the other types of sleep activities, thus leading to a coalescence of different rhythms that can only be observed in intact-brain animals and humans. Far from being epiphenomena, with no functional role, NREM sleep oscillations, particularly spindles and their experimental model augmenting responses, produce synaptic plasticity in target cortical neurons and resonant activity in corticothalamic loops, as in "memory" processes. Upon brain arousal, spindles are blocked by inhibition of RE neurons, the spindles' pacemakers; clock-like delta rhythm is obliterated by depolarization of TC neurons; and the cortically generated slow oscillation is abolished by selective erasure of its hyperpolarizing components. Fast (beta and gamma) oscillations are roduced by the depolarizing effects of mesopontine cholinergic neurons acting on TC neurons and nucleus basalis neurons acting on cortical neurons.


Determinants of virological failure and antiretroviral drug resistance in Mozambique.
The objective of this study was to inform public health actions to limit first-line ART failure and HIV drug resistance in Mozambique. This was a cross-sectional study. HIV-1-infected adults on first-line ART for at least 1 year attending routine visits in the Manhiça District Hospital, in a semi-rural area in southern Mozambique with no HIV-1 RNA monitoring available, were evaluated for clinical, socio-demographic, therapeutic, immunological and virological characteristics. Factors associated with HIV-1 RNA ≥1000 copies/mL and HIV drug resistance were determined using multivariate logistic regression. The study included 334 adults on first-line ART for a median of 3 years, of which 65% (214/332) had suppressed viraemia, 11% (37/332) had low-level viraemia (HIV-1 RNA 150-999 copies/mL) and 24% (81/332) had overt virological failure (HIV-1 RNA ≥1000 copies/mL). HIV drug resistance was detected in 89% of subjects with virological failure, but in none with low-level viraemia. Younger age [OR = 0.97 per additional year (95% CI = 0.94-1.00), P = 0.039], ART initiation at WHO stage III/IV [OR = 2.10 (95% CI = 1.23-3.57), P = 0.003] and low ART adherence [OR = 2.69 (95% CI = 1.39-5.19), P = 0.003] were associated with virological failure. Longer time on ART [OR = 1.55 per additional year (95% CI = 1.00-2.43), P = 0.052] and illiteracy [OR = 0.24 (95% CI = 0.07-0.89), P = 0.033] were associated with HIV drug resistance. Compared with HIV-1 RNA, clinician's judgement of ART failure, based on clinical and immunological outcomes, only achieved 29% sensitivity and misdiagnosed 1 out of every 4.5 subjects. Public health programmes in Mozambique should focus on early HIV diagnosis, early ART initiation and adherence support. Virological monitoring drastically improves the diagnosis of ART failure, enabling a better use of resources.


The role of new β-lactamase inhibitors in gram-negative infections.
In recent years, traditional β-lactams have dramatically reduced their effectiveness against gram-negative bacteria mainly because of their ability to express multiple β-lactamase or carabapenemases that are not hydrolyzed by the old β-lactam inhibitors (BLIs) such as clavulanic acid, tazobactam, and sulbactam. New BLIs molecules have been developed to face the need of compounds that are active against multidrug or pandrug resistant gram-negative pathogens. The aim of this review is to summarize the new generation of BLIs and β-lactams combinations. A number of new molecules with activity against Ambler class A (e.g., extended-spectrum β-lactamases, serine carbapenemases), class C (e.g., AmpC), or class D (e.g., oxacillinase-48) have been recently approved in combination with old β-lactams for the treatment of multidrug-resistant bacteria, and other agents are under investigation. These new compounds include diazabicyclooctanones non-β-lactam inhibitors (e.g., avibactam, relebactam, nacubactam) and boronic acid inhibitors (e.g., vaborbactam). Newly approved and investigational new BLIs are expected to offer many advantages for the management of patients with multidrug-resistant gram-negative pathogens. Promising characteristics of new compounds include high activity against multi drug resistance gram-negative bacteria and a favorable safety profile.


The RNA binding protein Quaking represses splicing of the Fibronectin EDA exon and downregulates the interferon response.
Quaking (QKI) controls RNA metabolism in many biological processes including innate immunity, where its roles remain incompletely understood. To illuminate these roles, we performed genome scale transcriptome profiling in QKI knockout cells with or without poly(I:C) transfection, a double-stranded RNA analog that mimics viral infection. Analysis of RNA-sequencing data shows that QKI knockout upregulates genes induced by interferons, suggesting that QKI is an immune suppressor. Furthermore, differential splicing analysis shows that QKI primarily controls cassette exons, and among these events, we noted that QKI silences splicing of the extra domain A (EDA) exon in fibronectin (FN1) transcripts. QKI knockout results in elevated production and secretion of FN1-EDA protein, which is a known activator of interferons. Consistent with an upregulation of the interferon response in QKI knockout cells, our results show reduced production of dengue virus-2 and Japanese encephalitis virus in these cells. In conclusion, we demonstrate that QKI downregulates the interferon system and attenuates the antiviral state.


Understanding the impacts of cellular environments on ligand binding of membrane receptors by computational simulations.
Binding of cell surface receptors with their extracellular ligands initiates various intracellular signaling pathways. However, our understanding of the cellular functions of these receptors is very limited due to the fact that in vivo binding between ligands and receptors has only been successfully measured in a very small number of cases. In living cells, receptors are anchored on surfaces of the plasma membrane, which undergoes thermal undulations. Moreover, it has been observed in various systems that receptors can be organized into oligomers prior to ligand binding. It is not well understood how these cellular factors play roles in regulating the dynamics of ligand-receptor interactions. Here, we tackled these problems by using a coarse-grained kinetic Monte Carlo simulation method. Using this method, we demonstrated that the membrane undulations cause a negative effect on ligand-receptor interactions. We further found that the preassembly of membrane receptors on the cell surface can not only accelerate the kinetics of ligand binding but also reduce the noises during the process. In general, our study highlights the importance of membrane environments in regulating the function of membrane receptors in cells. The simulation method can be potentially applied to specific receptor systems involved in cell signaling.


Evolution and impact of a dedicated ultrasound clinic on clinical rheumatology practice at an academic medical center.
Rheumatologic ultrasonography (RhUS) has grown in scope and application over the past 20 years. While many studies have shown the benefits of RhUS, few have investigated the efficacy of a dedicated clinic. This study explores the impact of a dedicated ultrasound clinic on patients and rheumatologists at an academic medical center (AMC). We analyzed claims data for patient visits, X-rays (XR), magnetic resonance imaging (MRI), and RhUS from an AMC with an established RhUS clinic, alongside two affiliated community medical practices (CMPs) without RhUS. We also analyzed RhUS clinic records on referral indication, procedures, results, and follow-up treatment changes. Pre- and post-RhUS visit patient surveys and referring physician (RP) surveys assessed experience and impact of the RhUS clinic. From 2018 to 2021, referrals to the RhUS clinic substantially increased. In parallel, XR and MRI orders changed by -76 % and -43 % respectively, compared with 163 % and -24 % at CMPs. Discordance between RP pre-RhUS assessments and RhUS results were common. Patient surveys showed RhUS led to increased disease understanding and impacted thoughts and decisions about their therapy. RPs found utility in RhUS across a range of indications and were confident with RhUS results. These findings suggest a dedicated RhUS clinic can be a valuable resource in clinical rheumatology practice. Implementation of a RhUS clinic at this AMC spurred rapid adoption of RhUS into clinical decision-making with notable benefits for patients and physicians alike. This may serve as a model for implementation of similar clinics at other institutions.


Facing the Realities of Pragmatic Design Choices in Environmental Health Studies: Experiences from the Household Air Pollution Intervention Network Trial.
Objective: Household Air Pollution Intervention Network (HAPIN) investigators tested a complex, non-pharmacological intervention in four low- and middle-income countries as a strategy to mitigate household air pollution and improve health outcomes across the lifespan. Intervention households received a liquefied petroleum gas (LPG) stove, continuous fuel delivery and regular behavioral reinforcements for 18 months, whereas controls were asked to continue with usual cooking practices. While HAPIN was designed as an explanatory trial to test the efficacy of the intervention on four primary outcomes, it introduced several pragmatic aspects in its design and conduct that resemble real-life conditions. We surveyed HAPIN investigators and asked them to rank what aspects of the design and conduct they considered were more pragmatic than explanatory. Methods: We used the revised Pragmatic Explanatory Continuum Indicator Summary (PRECIS-2) to survey investigators on the degree of pragmatism in nine domains of trial design and conduct using a five-point Likert rank scale from very explanatory (1) to very pragmatic (5). We invited 103 investigators. Participants were given educational material on PRECIS-2, including presentations, papers and examples that described the use and implementation of PRECIS-2. Results: Thirty-five investigators (mean age 42 years, 51% female) participated in the survey. Overall, only 17% ranked all domains as very explanatory, with an average (±SD) rank of 3.2 ± 1.4 across domains. Fewer than 20% of investigators ranked eligibility, recruitment or setting as very explanatory. In contrast, ≥50% of investigators ranked the trial organization, delivery and adherence of the intervention and follow-up as very/rather explanatory whereas ≤17% ranked them as rather/very pragmatic. Finally, <25% of investigators ranked the relevance of outcomes to participants and analysis as very/rather explanatory whereas ≥50% ranked then as rather/very pragmatic. In-country partners were more likely to rank domains as pragmatic when compared to investigators working in central coordination (average rank 3.2 vs. 2.8, respectively; Wilcoxon rank-sum p < 0.001). Conclusion: HAPIN investigators did not consider their efficacy trial to be rather/very explanatory and reported that some aspects of the design and conduct were executed under real-world conditions; however, they also did not consider the trial to be overly pragmatic. Our analysis underscores the importance of using standardized tools such as PRECIS-2 to guide early discussions among investigators in the design of environmental health trials attempting to measure efficacy.


Real-time neural signals of perceptual priming with unfamiliar geometric shapes.
Perceptual priming is a type of item-specific implicit memory that is distinct from explicit memory. Neural signals of the processing responsible for perceptual priming can be difficult to isolate due to concurrent conceptual processing and explicit recognition. We successfully identified neural correlates of perceptual priming by using minimally meaningful, difficult-to-recognize, kaleidoscope images. Human participants were required to quickly indicate the number of colors present in each stimulus, and priming was shown by faster and more accurate visual discriminations for repeated compared with initial presentations. Electroencephalographic responses linked with this differential perceptual fluency were identified as negative potentials 100-300 ms poststimulus onset. Furthermore, different potentials recorded during initial presentations were indicative of perceptual learning, in that their amplitude predicted the magnitude of later priming. These electrophysiological findings show that the degree of perceptual learning engaged upon first encountering a novel visual stimulus predicts the degree of perceptual fluency experienced when the stimulus is processed a second time. It is thus possible to isolate multiple neural processing stages relevant to perceptual priming by using real-time measures of relevant neurophysiological activity in conjunction with experimental circumstances that limit the contaminating influences of other neurocognitive events.


Proteomic prediction of incident heart failure and its main subtypes.
To examine the ability of serum proteins in predicting future heart failure (HF) events, including HF with reduced or preserved ejection fraction (HFrEF or HFpEF), in relation to event time, and with or without considering established HF-associated clinical variables. In the prospective population-based Age, Gene/Environment Susceptibility Reykjavik Study (AGES-RS), 440 individuals developed HF after their first visit with a median follow-up of 5.45 years. Among them, 167 were diagnosed with HFrEF and 188 with HFpEF. A least absolute shrinkage and selection operator regression model with nonparametric bootstrap were used to select predictors from an analysis of 4782 serum proteins, and several pre-established clinical parameters linked to HF. A subset of 8-10 distinct or overlapping serum proteins predicted different future HF outcomes, and C-statistics were used to assess discrimination, revealing proteins combined with a C-index of 0.80 for all incident HF, 0.78 and 0.80 for incident HFpEF or HFrEF, respectively. In the AGES-RS, protein panels alone encompassed the risk contained in the clinical information and improved the performance characteristics of prediction models based on NT-proBNP and clinical risk factors. Finally, the protein predictors performed particularly well close to the time of an HF event, an outcome that was replicated in the Cardiovascular Health Study (CHS). A small number of circulating proteins accurately predicted future HF in the AGES-RS cohort of older adults, and they alone encompass the risk information found in a collection of clinical data. Incident HF events were predicted up to eight years, with predictor performance significantly improving for events occurring less than one year ahead, a finding replicated in an external cohort study. This article is protected by copyright. All rights reserved.


Characterizing multistationarity regimes in biochemical reaction networks.
Switch like responses appear as common strategies in the regulation of cellular systems. Here we present a method to characterize bistable regimes in biochemical reaction networks that can be of use to both direct and reverse engineering of biological switches. In the design of a synthetic biological switch, it is important to study the capability for bistability of the underlying biochemical network structure. Chemical Reaction Network Theory (CRNT) may help at this level to decide whether a given network has the capacity for multiple positive equilibria, based on their structural properties. However, in order to build a working switch, we also need to ensure that the bistability property is robust, by studying the conditions leading to the existence of two different steady states. In the reverse engineering of biological switches, knowledge collected about the bistable regimes of the underlying potential model structures can contribute at the model identification stage to a drastic reduction of the feasible region in the parameter space of search. In this work, we make use and extend previous results of the CRNT, aiming not only to discriminate whether a biochemical reaction network can exhibit multiple steady states, but also to determine the regions within the whole space of parameters capable of producing multistationarity. To that purpose we present and justify a condition on the parameters of biochemical networks for the appearance of multistationarity, and propose an efficient and reliable computational method to check its satisfaction through the parameter space.


Voriconazole in the treatment of aspergillosis, scedosporiosis and other invasive fungal infections in children.
To describe the safety and efficacy of voriconazole in children treated within the compassionate release program. Children received voriconazole on a compassionate basis for treatment of an invasive fungal infection if they were refractory to or intolerant of conventional antifungal therapy. Voriconazole was administered as a loading dose of 6 mg/kg every 12 h i.v. on Day 1 followed by 4 mg/kg every 12 h i.v. thereafter. When feasible the route of administration of voriconazole was changed from i.v. to oral (100 or 200 mg twice a day for patients weighing < 40 or > or = 40 kg, respectively). Outcome was assessed by investigators at the end of therapy or at the last visit as success (complete or partial response), stable infection, or failure, based on protocol-defined criteria. Sixty-nine children (ages 9 months to 15 years; median, 7 years) received voriconazole; 58 had a proven or probable fungal infection. Among these 58 patients 27 had hematologic malignancies and 13 had chronic granulomatous disease as the most frequent underlying conditions. Forty-two patients had aspergillosis, 8 had scedosporiosis, 4 had invasive candidiasis and 4 had other invasive fungal infections. The median duration of voriconazole therapy was 93 days. At the end of therapy 26 patients (45%) had a complete or partial response. Four patients (7%) had a stable response, 25 (43%) failed therapy and 4 (7%) were discontinued from voriconazole because of intolerance. Success rates were highest in patients with chronic granulomatous disease (62%) and lowest in patients with hematologic malignancies (33%). Two patients experienced treatment-related serious adverse events (ulcerated lips with rash, elevated hepatic transaminases or bilirubin). A total of 23 patients had voriconazole-related adverse events, 3 (13%) of which caused discontinuation of voriconazole therapy. The most commonly reported adverse events included elevation in hepatic transaminases or bilirubin (n = 8), skin rash (n = 8), abnormal vision (n = 3) and a photosensitivity reaction (n = 3). These data support the use of voriconazole for treatment of invasive fungal infections in pediatric patients who are intolerant of or refractory to conventional antifungal therapy.


Sports injury and illness epidemiology during the 2014 Youth Olympic Games: United States Olympic Team Surveillance.
To describe injury and illness incidence from the US Youth Olympic Team during the 2014 Youth Olympic Games (YOG). Electronic health records of Team USA athletes registered for the YOG were reviewed for patient encounters during the 2014 YOG. Medical encounters were defined as all medical services provided by a healthcare provider including evaluation, treatment and prophylactic services. All medical conditions were categorised by IOC Injury and Illness reporting criteria. Team USA was comprised of 48 male and 46 female athletes, aged of 14-18 years, representing 20 sports. There were 346 total medical encounters among 54 of the 94 registered athletes for a rate of 3.7 medical encounters per athlete. A total of 40 injuries were recorded (14 time loss injuries, 26 non-time loss) and 20 illnesses. This equates to 43% of athletes sustaining an injury, and 21% sustaining an illness, with a frequency of 426 injuries and 213 illnesses per 1000 registered athletes. The most commonly involved body regions for time loss injuries were the knee (21%) and ankle (12%). Time loss injuries most frequently occurred in competition (71%) and the most common type of injury was a ligament sprain (43%). The most common illnesses were respiratory (35%) and dermatological conditions (30%). The rates of injury, illness and medical encounters per athlete were greater than in previous reports of medical service provisions at youth Olympic-level sporting events.


Detection of Polioviruses in Sewage Using Cell Culture and Molecular Methods.
The work presented here demonstrates the utility of a two-step algorithm for environmental poliovirus surveillance based on: preselection of sewage samples tested for the presence of enteroviral genetic material-RT-PCR assay and detection of infectious viruses by cell culture technique (L20B for polioviruses and RD for polio and other non-polio enteroviruses). RD and L20B cell lines were tested to determine their sensitivity for isolation of viruses from environmental samples (sewage). Finally, we wanted to determine if sewage concentration affects the results obtained for RT-PCR and cell cultures.


CD4+CD25+ Regulatory T Cells Inhibit Natural Killer Cell Hepatocytotoxicity of Hepatitis B Virus Transgenic Mice via Membrane-Bound TGF-β and OX40.
CD4+CD25+ regulatory T cells (Tregs) are involved in the regulation of physiological and pathological hepatic immune responses, but the roles are not well explored in natural killer (NK) cell-mediated liver diseases. In this study, using the NK cell-mediated oversensitive liver injury model of hepatitis B virus transgenic (HBs-Tg) mice triggered by a low dose of concanavalin A, it was observed that an increased number of CD4+CD25+Foxp3+ Tregs were accumulated in the liver, along with the recovery of liver injury. Adoptive transfer of hepatic Tregs from HBs-Tg mice but not wild B6 mice could significantly attenuate the oversensitive liver injury via inhibiting liver accumulation and decreasing NK cell group 2D-mediated activation of NK cells in the recipient HBs-Tg mice. Furthermore, upregulated expression of membrane-bound TGF-β (mTGF-β) and OX40 on hepatic Tregs were demonstrated to account for inhibiting the NK cell-mediated hepatic injury in HBs-Tg mice through cell-cell contact, confirmed by antibody blockade and cell Transwell experiments in vivo and in vitro. Our findings for the first time indicated that CD4+CD25+ Tregs directly suppressed NK cell-mediated hepatocytotoxicity through mTGF-β and OX40/OX40L interaction in a cell-cell contact manner in HBV-associated liver disease.


Embryo vaccination against Marek's disease with serotypes 1, 2 and 3 vaccines administered singly or in combination.
Marek's disease virus (MDV) vaccines of serotypes 1 and 2 administered in 18-day-old embryonated eggs induced better protection against post-hatch challenge at 3 days with virulent MDV than vaccines given at hatch. Embryonal vaccination with a polyvalent vaccine containing equal quantities of serotypes 1 and 2 of MDV and serotype 3 virus (turkey herpesvirus, HVT) was also significantly more effective than post-hatch vaccination. These and earlier results indicate that protective efficacy of single or combined Marek's disease vaccine serotypes against post-hatch challenge at 3 days can be substantially improved if the vaccines are injected into 18-day embryos rather than at hatch. Injection of vaccines of serotypes 1 or 2 into embryonated eggs or hatched chicks did not cause detectable gross or microscopic lesions in chickens. Vaccine viruses of serotypes 1 and 2 could be isolated from spleen cells of chickens 1 week post-vaccination, and the titer of recoverable viruses was higher in chickens that received the vaccines at the 18th day of embryonation than in chickens vaccinated at hatch. Although embryo vaccination with HVT usually provided better protection than post-hatch vaccination against early post-hatch challenge with variant pathotypes of MDV, the protection was poor regardless of vaccination protocol. If challenge with variant pathotypes of MDV was delayed until embryonally or post-hatch HVT-vaccinated chickens were 21 days of age, protection of chickens by HVT was not enhanced. Thus, resistance induced by embryonal vaccination with HVT was qualitatively similar to that induced by post-hatch vaccination with this virus.


Isolation and functional characterization of P-TEFb-associated factors that control general and HIV-1 transcriptional elongation.
Originally identified as a factor crucial for RNA polymerase (Pol) II transcriptional elongation of cellular genes, the P-TEFb kinase was subsequently shown to also serve as a specific host co-factor required for HIV-1 transcription. Recruited by either the bromodomain protein Brd4 to cellular promoters for general transcription or the HIV-1 Tat protein to the viral LTR for activated HIV-1 transcription, P-TEFb stimulates the processivity of Pol II through phosphorylating the C-terminal domain of Pol II and a pair of negative elongation factors, leading to the synthesis of full-length transcripts. However, abundant evidence indicates that P-TEFb does not act alone in the cell and that all of its known biological functions are likely mediated through the interactions with various regulators. Although a number of P-TEFb-associated factors have already been identified, there are likely more yet to be discovered. Given that P-TEFb plays an essential role in HIV-1 transcription, a major challenge facing the field is to identify all the P-TEFb-associated factors and determine how they may modulate Tat-transactivation and HIV-1 replication. Described here is a set of experimental procedures that have not only enabled us to isolate and identify several P-TEFb-associated factors, but also provided the means to characterize their biochemical functions in HIV-1 transcriptional control. In light of the recent demonstrations that transcriptional elongation plays a much more important role in controlling metazoan gene expression than previously thought, the techniques presented here will also be useful for analyzing Pol II elongation of cellular genes.


Vigorous physical activity and carotid distensibility in young and mid-aged adults.
Although physical activity (PA) improves arterial distensibility, it is unclear which type of activity is most beneficial. We aimed to examine the association of different types of PA with carotid distensibility (CD) and the mechanisms involved. Data included 4503 Australians and Finns aged 26-45 years. Physical activity was measured by pedometers and was self-reported. CD was measured using ultrasound. Other measurements included resting heart rate (RHR), cardiorespiratory fitness (CRF), blood pressure, biomarkers and anthropometry. Steps/day were correlated with RHR (Australian men r = -0.10, women r = - 0.14; Finnish men r = -0.15, women r = -0.11; P<0.01), CRF and biochemical markers, but not with CD. Self-reported vigorous leisure-time activity was more strongly correlated with RHR (Australian men r = -0.23, women r = -0.19; Finnish men r = -0.20, women r = -0.13; P < 0.001) and CRF, and was correlated with CD (Australian men r = 0.07; Finnish men r = 0.07, women r = 0.08; P < 0.05). This relationship of vigorous leisure-time activity with CD was mediated by RHR independently of potential confounders. In summary, vigorous leisure-time PA but not total or less intensive PA was associated with arterial distensibility in young to mid-aged adults. Promotion of vigorous PA is therefore recommended among this population. RHR was a key intermediary factor explaining the relationship between vigorous PA and arterial distensibility.


An overview of nanotoxicity and nanomedicine research: principles, progress and implications for cancer therapy.
The toxic paradigms of chemotherapeutic drugs and nanoparticles are tightly linked. Whereas uncontrolled exposure of living systems to therapeutics/nanomaterials leads to toxicity, selective induction of cytotoxicity in cancer cells helps in the fight against cancer. The increasing understanding of nanotoxicity paradigms has recently resulted in important benchmarks for the safe design of nanomaterial-based drug delivery systems aiming to fight cancer. In this context, this review aims to compile and present recent advances, outcomes and interconnections between nanomaterial-based drug delivery and nanotoxicity disciplines in order to provide a comprehensive guidance for future research. First, the basic concepts and mechanisms of nanomaterial-based drug delivery and nanotoxicity are introduced. Second, we present a detailed classification of drug delivery strategies and nanotoxicity paradigms, supported by the most recent research studies with a special focus on the interconnections between nanotoxicity and drug delivery research, which are highlighted in order to explore future opportunities for developing advanced therapeutic approaches. Finally, this review is concluded with future prospects on the use of nanoparticles for manipulating the behavior of cells and animals.


Probing the substrate specificity of the intracellular brain platelet-activating factor acetylhydrolase.
Platelet-activating factor acetylhydrolases (PAF-AHs) are unique PLA2s which hydrolyze the sn-2 ester linkage in PAF-like phospholipids with a marked preference for very short acyl chains, typically acetyl. The recent solution of the crystal structure of the alpha(1) catalytic subunit of isoform Ib of bovine brain intracellular PAF-AH at 1.7 A resolution paved the way for a detailed examination of the molecular basis of substrate specificity in this enzyme. The crystal structure suggests that the side chains of Thr103, Leu48 and Leu194 are involved in substrate recognition. Three single site mutants (L48A, T103S and L194A) were overexpressed and their structures were solved to 2.3 A resolution or better by X-ray diffraction methods. Enzyme kinetics showed that, compared with wild-type protein, all three mutants have higher relative activity against phospholipids with sn-2 acyl chains longer than an acetyl. However, for each of the mutants we observed an unexpected and substantial reduction in the V(max) of the reaction. These results are consistent with the model in which residues Leu48, Thr103 and Leu194 indeed contribute to substrate specificity and in addition suggest that the integrity of the specificity pocket is critical for the expression of full catalytic function, thus conferring very high substrate selectivity on the enzyme.


CX3CL1 up-regulation is associated with recruitment of CX3CR1+ mononuclear phagocytes and T lymphocytes in the lungs during cigarette smoke-induced emphysema.
CX3CR1 is expressed on monocytes, dendritic cells, macrophages, subsets of T lymphocytes, and natural killer cells and functions in diverse capacities such as leukocyte adhesion, migration, and cell survival on ligand binding. Expression of the CX3CL1 gene, whose expression product is the sole ligand for CX3CR1, is up-regulated in human lungs with chronic cigarette smoke-induced obstructive lung disease. At present, it is unknown whether CX3CL1 up-regulation is associated with the recruitment and accumulation of immune cells that express CX3CR1. We show that mice chronically exposed to cigarette smoke up-regulate CX3CL1 gene expression, which is associated with an influx of CX3CR1+ cells in the lungs. The increase in CX3CR1+ cells is primarily comprised of macrophages and T lymphocytes and is associated with the development of emphysema. In alveolar macrophages, cigarette smoke exposure increased the expression of both CX3CR1 and CX3CL1 genes. The inducibility of CX3CR1 expression was not solely dependent on a chronic stimulus because lipopolysaccharide up-regulated CX3CR1 in RAW264.7 cells in vitro and in mononuclear phagocytes in vivo. Our findings suggest a mechanism by which macrophages amplify and promote CX3CR1+ cell accumulation within the lungs during both acute and chronic inflammatory stress. We suggest that one function of the CX3CR1-CX3CL1 pathway is to recruit and sustain divergent immune cell populations implicated in the pathogenesis of cigarette smoke-induced emphysema.


On the evolutionary interplay between dispersal and local adaptation in heterogeneous environments.
Dispersal, whether in the form of a dandelion seed drifting on the breeze, or a salmon migrating upstream to breed in a nonnatal stream, transports genes between locations. At these locations, local adaptation modifies the gene frequencies so their carriers are better suited to particular conditions, be those of newly disturbed soil or a quiet river pool. Both dispersal and local adaptation are major drivers of population structure; however, in general, their respective roles are not independent and the two may often be at odds with one another evolutionarily, each one exhibiting negative feedback on the evolution of the other. Here, we investigate their joint evolution within a simple, discrete-time, metapopulation model. Depending on environmental conditions, their evolutionary interplay leads to either a monomorphic population of highly dispersing generalists or a collection of rarely dispersing, locally adapted, polymorphic sub-populations, each adapted to a particular habitat type. A critical value of environmental heterogeneity divides these two selection regimes and the nature of the transition between them is determined by the level of kin competition. When kin competition is low, at the transition we observe discontinuities, bistability, and hysteresis in the evolved strategies; however, when high, kin competition moderates the evolutionary feedback and the transition is smooth.


Metformin inhibits digestive proteases and impairs protein digestion in mice.
Metformin is among the most prescribed medications worldwide and the first-line therapy for type 2 diabetes. However, gastrointestinal side effects are common and can be dose limiting. The total daily metformin dose frequently reaches several grams, and poor absorption results in high intestinal drug concentrations. Here, we report that metformin inhibits the activity of enteropeptidase and other digestive enzymes at drug concentrations predicted to occur in the human duodenum. Treatment of mouse gastrointestinal tissue with metformin reduces enteropeptidase activity; further, metformin-treated mice exhibit reduced enteropeptidase activity, reduced trypsin activity, and impaired protein digestion within the intestinal lumen. These results indicate that metformin-induced protein maldigestion could contribute to the gastrointestinal side effects and other impacts of this widely used drug.


Cost-effectiveness analysis of ultrasonography screening for nonalcoholic fatty liver disease in metabolic syndrome patients.
Nonalcoholic fatty liver disease (NAFLD) can be diagnosed early by noninvasive ultrasonography; however, the cost-effectiveness of ultrasonography screening with intensive weight reduction program in metabolic syndrome patients is not clear. This study aims to estimate economic and clinical outcomes of ultrasonography in Thailand. Cost-effectiveness analysis used decision tree and Markov models to estimate lifetime costs and health benefits from societal perspective, based on a cohort of 509 metabolic syndrome patients in Thailand. Data were obtained from published literatures and Thai database. Results were reported as incremental cost-effectiveness ratios (ICERs) in 2014 US dollars (USD) per quality-adjusted life year (QALY) gained with discount rate of 3%. Sensitivity analyses were performed to assess the influence of parameter uncertainty on the results. The ICER of ultrasonography screening of 50-year-old metabolic syndrome patients with intensive weight reduction program was 958 USD/QALY gained when compared with no screening. The probability of being cost-effective was 67% using willingness-to-pay threshold in Thailand (4848 USD/QALY gained). Screening before 45 years was cost saving while screening at 45 to 64 years was cost-effective. For patients with metabolic syndromes, ultrasonography screening for NAFLD with intensive weight reduction program is a cost-effective program in Thailand. Study can be used as part of evidence-informed decision making. Findings could contribute to changes of NAFLD diagnosis practice in settings where economic evidence is used as part of decision-making process. Furthermore, study design, model structure, and input parameters could also be used for future research addressing similar questions.


Hip Instability in Patients With Down Syndrome.
The incidence of hip instability in children with Down syndrome is 1% to 7%. The natural history is often progressive, with the typical onset of hypermobility of the hip evolving to habitual dislocation, persistent subluxation, and fixed dislocation, and eventually leading to the loss of independent mobility. Treatment focuses on stabilizing the hip joint and depends on the patient's age and the severity of the disease. Typically, surgical intervention is recommended for the treatment of patients with habitual dislocation, subluxation, and complete dislocation of the hip. When indicated, surgical management must take into account associated anatomic abnormalities of the femur and acetabulum. Hip instability in Down syndrome may persist despite surgical intervention and remains a difficult condition to manage.


Computational fluid dynamics of the ascending aorta before the onset of type A aortic dissection.
We performed a pre-dissection computational fluid analysis of an ascending aortic aneurysm associated with unicuspid aortic valve. The analysis showed an abnormal helical flow pattern inside the aneurysm and an increased wall stress on the right postero-lateral wall of the ascending aorta. These values were largely higher than the theoretical cut-off for aortic wall dissection, their topographic distribution followed the intimal tear site as subsequently diagnosed by computed tomography scan and confirmed during the operation for dissection repair.


Protective effects of ginsenoside Re on lipopolysaccharide-induced cardiac dysfunction in mice.
The impaired cardiac function caused by reduced myocardial contractility is a typical manifestation of sepsis/septic shock. Ginsenoside Re (GS-Re) is one of the most abundant ingredients of ginseng. This study was designed to investigate the protective effects of GS-Re on lipopolysaccharide (LPS)-induced septic cardiac dysfunction and inflammatory response in mice. Mice were intragastrically administered with GS-Re (15 mg kg(-1)) for 1 week before the LPS challenge (10 mg kg(-1), i.p.). Cardiac function was evaluated 6 h after LPS induction. GS-Re pretreatment significantly protected against LPS-induced cardiac dysfunction. GS-Re ameliorated the imbalance between iNOS and eNOS, and prevented NF-κB activation and subsequent myocardial inflammatory responses in endotoxemic mice. The effects of GS-Re were closely associated with estrogen receptors (ERs), phosphatidylinositide 3-kinase (PI3K)/protein kinase B (AKT) signaling, and the mitogen-activated protein kinase signaling pathway, as characterized by the GS-Re-induced preservation of ERα, ERβ, and phospho-Akt and inhibition of phospho-ERK1/2, phospho-JNK, phospho-P38. However, GS-Re had no effect on LPS-induced activation of TLR-4. All these results showed that GS-Re pretreatment significantly attenuated LPS-induced cardiac dysfunction and inflammatory response.


Borreliacidal OspC antibodies specific for a highly conserved epitope are immunodominant in human lyme disease and do not occur in mice or hamsters.
Humans produce highly specific borreliacidal antibodies against outer surface protein C (OspC) shortly after infection with Borrelia burgdorferi sensu stricto. We previously demonstrated the epitope recognized by immunoglobulin M (IgM) and IgG OspC borreliacidal antibodies was located within the 50 amino acids nearest the carboxy (C) terminus. In this study, we show the immunodominant epitope is located in the highly conserved region within the seven C-terminal amino acids. Six early Lyme disease sera that contained borreliacidal activity and IgM and/or IgG OspC antibodies were chosen randomly and adsorbed with truncated OspC containing the 16 or 7 amino acids nearest the C terminus. Adsorptions with each truncated protein abrogated the borreliacidal activity completely. In addition, only small concentrations of OspC antibodies remained detectable by enzyme-linked immunosorbent assay and Western blotting. Moreover, borreliacidal OspC antibodies were not induced in laboratory mice or hamsters despite heavy infections with B. burgdorferi spirochetes. These findings confirm that borreliacidal antibodies comprise the majority of the IgM and IgG OspC antibody response in human Lyme disease and that the epitope is located in the highly conserved C terminus. In addition, rodent animal models appear to be inappropriate subjects for assessing the effectiveness of the epitope for serodiagnosis or as a human Lyme disease vaccine.


Activated Stat related transcription factors in acute leukemia.
Cell proliferation and differentiation are under the control of cytokines and growth factors. Different signaling pathways are involved in the transmission of a specific signal through successive phosphorylation and dephosphorylation of proteins leading to gene transcription necessary for growth and differentiation. The cytokines and growth factors activate the Stat family of transcription factors. The Jak-Stat pathway is essential for cytokine signal transduction. Dysregulation of this cascade might lead to uncontrolled hematopoiesis. Studies have been carried out to examine the functionality of this pathway in cells from patients with acute leukemia. Members of the Stat protein family (Stat1, Stat3 and Stat5) are constitutively activated in cells collected from some acute leukemias suggesting dysregulation of the Jak-Stat pathway. Evidence of the existence of constitutively activated spliced variants of Stat3 and Stat5 proteins are described. The mechanisms of such activation remain to be clarified.


Isolation and characterization of antioxidant protein fractions from melinjo (Gnetum gnemon) seeds.
The protein from the seeds of melinjo ( Gnetum gnemon ) was purified using a precipitation method and ion exchange chromatographic techniques to identify the potent antioxidant and free radical scavenging activities. Two antioxidant protein fractions were isolated from G. gnemon seed with molecular weights of approximately 30 kDa (Gg-AOPI) and 12 kDa (Gg-AOPII) by SDS-PAGE. The N-terminal amino acid sequence of Gg-AOPII is Gly-Asn-Gly-Lys-Ala-Thr-Val-Ala-Ile-Leu-Val-Lys-Glu-Lys-Val-Glu-Tyr-Gly-Glu-Glu, and the result of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) analysis showed that they were distinct from each other; no protein in database matching was found to both Gg-AOPI and Gg-AOPII. The antioxidant or free radical scavenging activities of Gg-AOPs were investigated by employing in vitro assay systems including the inhibition of linoleic acid autoxidation, scavenging effect on α,α-diphenyl-β-picrylhydrazyl free radical (DPPH), 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), reducing power, chelating abilities of metal ions Cu(2+) and Fe(2+), and protections against hydroxyl radical-mediated DNA damages. The result showed that two protein fractions exhibited significant (p < 0.05) antioxidant activities against free radicals such as DPPH, ABTS, and superoxide anion and showed activities similar to those of glutathione (G-SH) and BHT in a linoleic acid emulsion assay system. Moreover, Gg-AOPI and Gg-AOPII also exhibited notable reducing power and strong chelating effect on Fe(2+) and protected hydroxyl radical induced oxidative DNA damage. The data obtained by the in vitro systems obviously established the antioxidant potency of Gg-AOPs.


Examining the Cognitive Proficiency Index in rehabilitation patients.
This study examined the Wechsler Adult Intelligence Scale-Fourth Edition (WAIS-IV) Cognitive Proficiency Index (CPI) in relation to other WAIS-IV indices, overall test battery mean (OTBM), and impairment (IMP) in an outpatient rehabilitation setting. Participants (N = 329) were 35% female and 88% Caucasian with average age and education of 42.9 (SD = 13.5) and 13.6 (SD = 2.4) years, respectively. Participants were grouped by diagnosis and validity: traumatic brain injury (TBI; n = 176; 39% mild), cerebrovascular accident (CVA; n = 52), other neurologic and psychiatric conditions (OTH; n = 49), and questionable performance validity (QPV; n = 52). OTBM was calculated from non-WAIS-IV tests; IMP was dichotomously defined as four or more non-WAIS-IV scores below cutoff (≤35 T). Significant group differences were observed on CPI, WAIS-IV indices, OTBM, and IMP. CPI significantly contributed (β = .51) to a linear regression model predicting OTBM (R2 = .63) with education and GAI as covariates. A logistic regression model with IMP as the outcome and education, GAI, and CPI as predictors correctly classified 80% of cases with area under the curve of .86. A previously identified cutoff (CPI < 84) correctly classified 65-78% of clinical groups categorized by IMP. A novel cutoff (CPI ≤ 80) differentiated clinical participants with history of mild TBI from the QPV group with sensitivity of 44.2% and specificity of 89.7%. CPI showed incremental validity in predicting OTBM and IMP and warrants further study as a useful clinical addition to other WAIS-IV indices.


[Public health risk threats associated with coronavirus infection in Moscow and the need to counteract].
to determine the main threats to the health of the population of Moscow based on the analysis of the medical and demographic situation in Moscow and the Russian Federation in the context of the COVID-19 pandemic, in order to develop management decisions at the regional level. Materials of the Federal State Statistics Service (Rosstat), as well as statistical collections of the Russian Research Institute of Health of the Ministry of Health of Russia for 2013-2022 were used. Coronavirus infection has changed the morbidity and mortality rates of the population of the Russian Federation and its constituent entities. In the Russian Federation mortality in 2019-2021 increased from 12.3 to 13.6‰, while in Moscow the increase in the indicator is significant - from 9.5 to 16.7‰. The incidence of COVID-19 in Moscow in 2020-2021 (over the two COVID years) increased from 6191.0 to 8976.0‰oo. The noted decrease in incidence rates across almost all classes of diseases during the COVID period (2019-2020) was associated with the lower medical check-ups and preventive activities, while during 2019-2021 there was an increase in disease incidence of the following classes: mental disorders, behavior disorders; diseases of the blood and blood-forming organs and certain disorders involving the immune mechanism, etc., indicating the need for rehabilitation of this group of patients. The need to counter the threats to the health of the population of Moscow caused by the coronavirus infection requires the development of management decisions aimed at strengthening preventive activities and rehabilitation of patients in the post-COVID period.


A simple validated clinical tool to predict the absence of coronary artery disease in patients with systolic heart failure of unclear etiology.
Coronary artery disease (CAD) is a major cause of systolic heart failure (HF). Identifying CAD as a cause of systolic HF has prognostic and treatment implications. Whether all patients with systolic HF of unclear etiology should undergo coronary angiography has been controversial. We sought to derive and validate a clinical prediction rule to exclude CAD as a cause of systolic HF. A derivation cohort was formed of consecutive patients who had undergone coronary angiography with a primary diagnosis of systolic HF of unclear etiology (ejection fraction <50%). Using multivariate logistic regression analysis, we derived a prediction rule for severe CAD (≥50% diameter stenosis in the left main, 3-vessel CAD, and 2-vessel CAD involving the proximal left anterior descending artery). The diagnostic performance of the defined prediction rule was prospectively validated in a separate cohort recruited from 2 institutions. Of the 124 patients in the derivation cohort, 27% had CAD, including 15% with severe CAD. The independent predictors of severe CAD included diabetes (odds ratio 5.1, p = 0.005), electrocardiographic Q waves or left bundle branch block (odds ratio 3.8, p = 0.02), and ≥2 nondiabetes risk factors: age (men ≥55 or women ≥65 years), dyslipidemia, hypertension, and tobacco use (odds ratio 4.8, p = 0.02). A prediction rule of having ≥1 independent predictor identified 97% of the patients with CAD and 100% of the patients with severe CAD. In the prospective validation cohort of 143 patients, the prediction rule had 98% sensitivity and 18% specificity for CAD but 100% sensitivity for severe CAD. In conclusion, a simple clinical prediction rule can accurately identify patients with CAD and eliminate the need for angiography in a substantial proportion of patients with systolic HF, with potentially significant cost savings and risk avoidance.


Regimens of less than six months for treating tuberculosis.
WHO recommends 6 months of treatment in TB programmes. The purpose of this review is to assess the effects of regimens lasting less than 6 months compared with longer regimens in the treatment of active TB. Search strategy: MEDLINE 1955-, Cochrane Infectious Diseases Trials Register, existing reviews, and researchers in the field. Date of the most recent search: January 1999. Randomized trials comparing two or more TB drug regimens, in which at least one regimen was <6 months and it was compared with at least one regimen that lasted longer, in any patients with active TB. One reviewer extracted data and assessed trial quality. Seven trials with a total of 9 comparisons of <6 months (range: 2-5 months) versus longer treatment were included. About 2200 patients were in the shorter regimens and about 1900 in the longer regimens (the same comparison groups were used for more than one shorter regimen, in two studies). Relapse rates were consistently higher after shorter duration treatment regimens, regardless of the comparison made, though they were all relatively low. Results were significantly better in the longer groups in the meta-analyses of 2, 3, and 4 months of treatment vs longer treatment (Peto OR = 6.1 [95%CI 2.19,17.01], 3.67 [2.42,5.58], 3.64 [1.71,7. 75] but not in the single trial of 5 vs. 7 months (Peto OR = 2.24 [0. 90,5.59]. Relapse rates after longer (comparison) regimens ranged from 0-7% at one year (or more), and in the shorter treatment arms, they ranged from 1-9% in 8 trials, and18% relapsed in the one remaining. There was little or no difference in the rates of adverse reactions or toxicity requiring a change of regimen or discontinuation of treatment. The "sterilizing efficacy" at the end of treatment varied little among treatments, providing no predictive value for relapse rates. Few or no deaths were reported in the individual trials, and in no case did enough deaths occur for a comparison of short vs. long regimens. Longer periods of treatment (at least up to 6 months) result in higher success rates in patients with active TB, but the differences are small. Under field conditions, where adherence to treatment is a big problem, and shorter regimens might improve adherence, these differences may not be evident. A comparison of <6 months vs. 6 months of treatment under programme conditions would be needed to determine this.


Mannitol, a novel bacterial compatible solute in Pseudomonas putida S12.
The aim of this study was to identify the compatible solutes accumulated by Pseudomonas putida S12 subjected to osmotic stress. In response to reduced water activity, P. putida S12 accumulated Nalpha-acetylglutaminylglutamine amide (NAGGN) simultaneously with a novel compatible solute identified as mannitol (using 13C- and 1H-nuclear magnetic resonance, liquid chromatography-mass spectroscopy and high-performance liquid chromatography methods) to maximum concentrations of 74 and 258 micromol g (dry weight) of cells(-1), respectively. The intracellular amounts of each solute varied with both the type and amount of osmolyte applied to induce osmotic stress in the medium. Both solutes were synthesized de novo. Addition of betaine to the medium resulted in accumulation of this compound and depletion of both NAGGN and mannitol. Mannitol and NAGGN were accumulated when sucrose instead of salts was used to reduce the medium water activity. Furthermore, both compatible solutes were accumulated when glucose was substituted by other carbon sources. However, the intracellular quantities of mannitol decreased when fructose, succinate, or lactate were applied as a carbon source. Mannitol was also raised to high intracellular concentrations by other salt-stressed Pseudomonas putida strains. This is the first study demonstrating a principal role for the de novo-synthesized polyol mannitol in osmoadaptation of a heterotrophic eubacterium.


A nationwide population-based study of migraine in Brazil.
The aim of this study was to estimate the 1-year prevalence of migraine and the degree of the association of migraine with some sociodemographic characteristics of a representative sample of the adult population of Brazil. This was a cross-sectional, population-based study. Telephone interviews were conducted on 3848 people, aged 18-79 years, randomly selected from the 27 States of Brazil. The estimated 1-year gender- and age-adjusted prevalence of migraine was 15.2%. Migraine was 2.2 times more prevalent in women, 1.5 times more in subjects with > 11 years of education, 1.59 times more in subjects with income of < 5 Brazilian Minimum Wages per month, and 1.43 times more in those who do not do any physical exercise. The overall prevalence of migraine in Brazil is 15.2%. Migraine is significantly more prevalent in women, subjects with higher education, with lower income, and those who do not exercise regularly, independently of their body mass index.


Delivery levels and behavior of 1,3-butadiene, acrylonitrile, benzene, and other toxic volatile organic compounds in mainstream tobacco smoke from two brands of commercial cigarettes.
Mainstream tobacco smoke (MTS) was collected from Camel and Marlboro cigarettes for the determination of the delivery levels and equilibrium gas/particle partitioning constants K(p) (m(3) microg(-)(1)) of 26 volatile organic compounds (VOCs) of toxicological interest. K(p) values are important for understanding the fractional distribution of each compound of interest between the gas and the particle phases of MTS. The experimental method involved (i) drawing a smoke sample into a Teflon sampling bag at 20 degrees C, (ii) allowing the smoke particulate matter (PM) to collect on the walls of the bag, (iii) sampling the bag to determine the initial gas phase concentration of each VOC, (iv) removing the gas phase from the bag, (v) refilling the bag with humidified nitrogen gas, (vi) reestablishing the gas/PM equilibrium, and (vii) redetermining the gas phase concentrations. For each smoke sample, a comparison of the initial and redetermined gas phase concentrations allowed calculation of the total (i.e., gas + particle) delivery level (= m(tot), ng cig(-)(1)) and K(p) value (= c(p)/c(g)) at 20 degrees C for each compound, where c(p) (ng microg(-)(1)) = concentration in the PM phase and c(g) (ng m(-)(3)) = concentration in the gas phase. Significant deliveries were observed for a number of carcinogenic VOCs. For the Camel cigarettes tested, the average m(tot) values for 1,3-butadiene, acrylonitrile, and benzene were 10(4.6), 10(4.4), and 10(4.8) ng cig(-)(1), respectively; for Marlboro, the m(tot) values were 10(5.0), 10(4.6), and 10(4.7) ng cig(-)(1), respectively. For each of the 26 VOCs, the smoke PM from the two brands yielded very similar K(p) values at 20 degrees C. In addition, the vapor pressure-dependent K(p) values of the 26 VOCs were in close agreement with predictions made by the Pankow theory of absorptive gas/particle partitioning [Pankow, J. F. (1994) Atmos. Environ. 28, 185-188]. These results can be used in general predictions of chemical behavior in tobacco smoke, including deposition mechanisms and rates in the respiratory tract from inhaled MTS. Example calculations are provided to illustrate how the gas phase fraction at equilibrium (f(g,e)) increases strongly with increasing compound vapor pressure and temperature and with dilution of the inhaled tobacco smoke total PM concentration (microg m(-)(3)).


Floristic diversity of regenerated tree species in Dipterocarp forests in Western Ghats of Karnataka, India.
The research was focused on exploring the structure, diversity and form of regeneration process of the Dipterocarp forests in Western Ghats in relation to environmental factors. Eight populations in the distribution range of Dipterocarp forests were selected. In each population 32 plots of 2mx2m were laid down randomly. Atotal of 1243 seedlings < or = 10 cm dbh (diameter at breast height) belonging to 99 species and 48 families were recorded. The number of regenerated tree species was found to be high in the populations of Mudigere (40), Sakleshpura (40) and Makuta (39), which are characterized by favorable locality factors and lower disturbances. The highest similarity index in species composition was recorded between the populations of Sampaje in Kodagu district and Gundya in Dakshina Kannada (60%) whereas the lowest similarity index was observed between the population of Sringeri in Chikmagalore and Sampaje (53%) and Gundya and Makuta (35%) in Kodagu district. Dipterocarpus indicus was found to be dominant among the regenerated tree species in all the sites studied except Gundy and Sampaje. The frequencies of regeneration classes (seedlings, saplings, poles and adult trees) were shaped as inverse J curve indicating the normal regeneration pattern under the present disturbance. The average disturbance of litter collection, grazing, fire, weeds and canopy opening were significant among different populations (p < or = 0.05). Negative correlation was observed between disturbance and species richness, number of individuals and density.


Biliary carcinoembryonic antigen levels in diagnosis of occult hepatic metastases from colorectal carcinoma.
To prospectively explore the role of carcinoembryonic antigen (CEA) in gallbladder bile in patients with colorectal carcinoma and the morphological and clinical features of neoplasia and the occurrence of hepatic metastases. CEA levels in the gallbladder and peripheral blood were studied in 44 patients with colorectal carcinoma and 10 patients with uncomplicated cholelithiasis. CEA samples were collected from the gallbladder bile and peripheral blood during the operation, immediately before extirpating the colorectal neoplasia or cholecystectomy. Values of up to 5 ng/ml were considered normal for bile and serum CEA. In the 44 patients with colorectal carcinoma who underwent operation with curative intent, the average level of serum CEA was 8.5 ng/ml (range: 0.1 to 111.0 ng/ml) and for bile CEA it was 74.5 ng/ml (range: 0.2 to 571.0 ng/ml). In the patients with uncomplicated cholelithiasis who underwent cholecystectomy, the average level of serum CEA was 1.9 ng/ml (range: 1.0 to 3.5 ng/ml) and for bile CEA it was 1.2 ng/ml (range: 0.3 to 2.9 ng/ml). The average duration of follow-up time was 16.5 months (range: 6 to 48 months). Four patients who underwent extirpation of the colorectal carcinoma without evidence of hepatic metastasis and with an average bile CEA value of 213.2 ng/ml presented hepatic metastases between three and seventeen months after removal of the primary colorectal neoplasia. Three of them successfully underwent extirpation of the hepatic lesions. High CEA levels in gallbladders of patients undergoing curative operation for colorectal carcinoma may indicate the presence of hepatic metastases. Such patients must be followed up with special attention to the diagnosis of such lesions.


Migration of human neutrophils in hydrated collagen lattices.
The migration of human neutrophilic granulocytes in hydrated collagen lattices was studied by a combination of cinemicroscopy, and scanning and transmission electron microscopy. The basic pattern of cell migration in collagen was similar to that observed previously for these cells on inert material surfaces; i.e., a cycle of cell extension and cytoplasmic flow into the leading extension. In general, however, neutrophils in collagen were less spread than on glass or plastic surfaces. Thin lamellipodia were absent and the leading extension of the cells was often an elaborately folded pseudopodium. In addition, neutrophils migrating in collagen were never observed to have retraction fibres at the tail end of the cells, although a uropod was usually seen. In the region of the uropod, extensive blebbing of the cells often occurred, and when this happened, forward movement of the cells ceased. At the ultrastructural level, both the leading pseudopodium and the blebs at the tail of the cell were found to contain a dense cytoskeletal network from which cell organelles were excluded. Finally, the cells were found to be coated with an extensive glycocalyx, and individual collagen fibres were sometimes observed within the glycocalyx.


The impact of NHS Health Checks on the prevalence of disease in general practices: a controlled study.
NHS Health Checks is a national case-finding and vascular risk assessment programme in England. No research has been published to assess the impact of NHS Health Checks on the prevalence of chronic disease in GP practices. To examine the impact of NHS Health Checks on the prevalence of hypertension, coronary heart disease (CHD), chronic kidney disease (CKD), atrial fibrillation (AF), and diabetes within practices, and compare this with usual medical care. A non-randomised controlled study in a mixed rural and urban county in England. Thirty-eight GP practices provided NHS Health Checks over a 3-year period. Forty-one practices that did not provide Health Checks acted as controls. t-tests and multiple linear regression were used to assess the difference in prevalence of disease between intervention group and control group practices, and the impact of NHS Health Checks on this. Throughout the duration of the study, 1142 previously undiagnosed cases of disease were detected through a total of 16 669 NHS Health Checks. Despite this, there were no significant differences in the change to the prevalence of diabetes, hypertension, CHD, CKD, and AF in practices providing NHS Health Checks compared with control practices. Regression analysis did not demonstrate that there was any significant association between the proportion of the eligible population of a practice having completed NHS Health Checks and changes in the prevalence of the five conditions studied. In practices providing NHS Health Checks, the change in the reported prevalence of diabetes, hypertension, CHD, CKD, and AF did not differ from that of practices providing usual care.


Osteocalcin protects against nonalcoholic steatohepatitis in a mouse model of metabolic syndrome.
Nonalcoholic fatty liver disease, particularly its more aggressive form, nonalcoholic steatohepatitis (NASH), is associated with hepatic insulin resistance. Osteocalcin, a protein secreted by osteoblast cells in bone, has recently emerged as an important metabolic regulator with insulin-sensitizing properties. In humans, osteocalcin levels are inversely associated with liver disease. We thus hypothesized that osteocalcin may attenuate NASH and examined the effects of osteocalcin treatment in middle-aged (12-mo-old) male Ldlr(-/-) mice, which were fed a Western-style high-fat, high-cholesterol diet for 12 weeks to induce metabolic syndrome and NASH. Mice were treated with osteocalcin (4.5 ng/h) or vehicle for the diet duration. Osteocalcin treatment not only protected against Western-style high-fat, high-cholesterol diet-induced insulin resistance but substantially reduced multiple NASH components, including steatosis, ballooning degeneration, and fibrosis, with an overall reduction in nonalcoholic fatty liver disease activity scores. Further, osteocalcin robustly reduced expression of proinflammatory and profibrotic genes (Cd68, Mcp1, Spp1, and Col1a2) in liver and suppressed inflammatory gene expression in white adipose tissue. In conclusion, these results suggest osteocalcin inhibits NASH development by targeting inflammatory and fibrotic processes.


The response of coarse root biomass to long-term CO2 enrichment and nitrogen application in a maturing Pinus taeda stand with a large broadleaved component.
Elevated atmospheric CO2 (eCO2 ) typically increases aboveground growth in both growth chamber and free-air carbon enrichment (FACE) studies. Here we report on the impacts of eCO2 and nitrogen amendment on coarse root biomass and net primary productivity (NPP) at the Duke FACE study, where half of the eight plots in a 30-year-old loblolly pine (Pinus taeda, L.) plantation, including competing naturally regenerated broadleaved species, were subjected to eCO2 (ambient, aCO2 plus 200 ppm) for 15-17 years, combined with annual nitrogen amendments (11.2 g N m-2 ) for 6 years. Allometric equations were developed following harvest to estimate coarse root (>2 mm diameter) biomass. Pine root biomass under eCO2 increased 32%, 1.80 kg m-2 above the 5.66 kg m-2 observed in aCO2 , largely accumulating in the top 30 cm of soil. In contrast, eCO2 increased broadleaved root biomass more than twofold (aCO2 : 0.81, eCO2 : 2.07 kg m-2 ), primarily accumulating in the 30-60 cm soil depth. Combined, pine and broadleaved root biomass increased 3.08 kg m-2 over aCO2 of 6.46 kg m-2 , a 48% increase. Elevated CO2 did not increase pine root:shoot ratio (average 0.24) but increased the ratio from 0.57 to 1.12 in broadleaved species. Averaged over the study (1997-2010), eCO2 increased pine, broadleaved and total coarse root NPP by 49%, 373% and 86% respectively. Nitrogen amendment had smaller effects on any component, singly or interacting with eCO2 . A sustained increase in root NPP under eCO2 over the study period indicates that soil nutrients were sufficient to maintain root growth response to eCO2 . These responses must be considered in computing coarse root carbon sequestration of the extensive southern pine and similar forests, and in modelling the responses of coarse root biomass of pine-broadleaved forests to CO2 concentration over a range of soil N availability.


Three-dimensional-object recognition by use of single-exposure on-axis digital holography.
On-axis phase-shifting digital holography requires recording of multiple holograms. We describe a novel real-time three-dimensional- (3-D-) object recognition system that uses single-exposure on-axis digital holography. In contrast to 3-D-object recognition by means of a conventional phase-shifting scheme that requires multiple exposures, our proposed method requires only a single digital hologram to be synthesized and used to recognize 3-D objects. A benefit of the proposed 3-D recognition method is enhanced practicality of digital holography for 3-D recognition in terms of its simplicity and greater robustness to external scene parameters such as moving targets and environmental noise factors. We show experimentally the utility of the single-exposure on-axis digital holography-based 3-D-object recognition method.


Pain Management After Hip Arthroscopy: Systematic Review of Randomized Controlled Trials and Cohort Studies.
Hip arthroscopy is often associated with significant postoperative pain and opioid-associated side effects. Effective pain management after hip arthroscopy improves patient recovery and satisfaction and decreases opioid-related complications. To collect, examine, and provide a comprehensive review of the available evidence from randomized controlled trials and comparative studies on pain control after hip arthroscopy. Systematic review. Using the Preferred Reporting Items for Systematic Reviews and Meta-analyses guidelines, a systematic review of the literature for postoperative pain control after hip arthroscopy was performed using electronic databases. Only comparative clinical studies with level 1 to 3 evidence comparing a method of postoperative pain control with other modalities or placebo were included in this review. Case series and studies without a comparative cohort were excluded. Several methods of pain management have been described for hip arthroscopy. A total of 14 studies met our inclusion criteria: 3 on femoral nerve block, 3 on lumbar plexus block, 3 on fascia iliaca block, 4 on intra-articular injections, 2 on soft tissue surrounding surgical site injection, and 2 on celecoxib (4 studies compared 2 or more methods of analgesia). The heterogeneity of the studies did not allow for pooling of data. Single-injection femoral nerve blocks and lumbar plexus blocks provided improved analgesia, but increased fall rates were observed. Fascia iliaca blocks do not provide adequate pain relief when compared with surgical site infiltration with local anesthetic and are associated with increased risk of cutaneous nerve deficits. Patients receiving lumbar plexus block experienced significantly decreased pain compared with fascia iliaca block. Portal site and periacetabular injections provide superior analgesia compared with intra-articular injections alone. Preoperative oral celecoxib, compared with placebo, resulted in earlier time to discharge and provided significant pain relief up to 24 hours. Perioperative nerve blocks provide effective pain management after hip arthroscopy but must be used with caution to decrease risk of falls. Intra-articular and portal site injections with local anesthetics and preoperative celecoxib can decrease opioid consumption. There is a lack of high-quality evidence on this topic, and further research is needed to determine the best approach to manage postoperative pain and optimize patient satisfaction.


Ensuring equity: Pharmacogenetic implementation in rural and tribal communities.
Implementation strategies for pharmacogenetic testing have been largely limited to major academic medical centers and large health systems, threatening to exacerbate healthcare disparities for rural and tribal populations. There exists a need in Montana (United States)-a state where two-thirds of the population live in rural areas and with a large proportion of tribal residents-to develop novel strategies to make pharmacogenetic testing more broadly available. We established partnerships between University of Montana (UM) and three early adopter sites providing patient-centered care to historically neglected populations. We conducted 45 semi-structured interviews with key stakeholders at each site and solicited participant feedback on the utility of a centralized pharmacogenetic service at UM offering consultations to patients and providers statewide via telehealth. For settings serving rural patients-tribal and non-tribal-participants described healthcare facilities without adequate infrastructure, personnel, and funding to implement pharmacogenetic services. Participants serving tribal communities stressed the need for ethical practices for collecting biospecimens and returning genetic results to patients, largely due to historical and contemporary traumas experienced by tribal populations with regard to genetic research. Participants expressed that pharmacogenetic testing could benefit patients by achieving therapeutic benefit sooner, reducing the risk of side effects, and improving adherence outcomes for patients with limited access to follow-up services in remote areas. Others expressed concern that financial barriers to pharmacogenetic testing for patients of lower socioeconomic status would further exacerbate inequities in care. Participants valued the role of telehealth to deliver pharmacogenetic consults from a centralized service at UM, describing the ability to connect providers and patients to resources and expertise as imperative to driving successful pharmacogenetic implementation. Our results support strategies to improve access to pharmacogenetic testing for neglected patient populations and create opportunities to reduce existing healthcare inequities. By exploring critical challenges for pharmacogenetic implementation focused on serving underserved communities, this work can help guide equitable frameworks to serve as a model for other resource-limited settings looking to initiate pharmacogenetic testing.


Expression of the SOS genes of Escherichia coli in Salmonella typhimurium.
To lysogenize Salmonella typhimurium by Lambda phage, a region of 10.2 kb of Escherichia coli DNA carrying the nusA gene was cloned in a S. typhimurium strain containing a F'112 plasmid which codifies for the lamB region of E. coli. The strain of S. typhimurium obtained in this way, was lysogenized by lambda c IndO- bacteriophage harboring either a fusion between recA1 or sfiA genes of E. coli with lacZ gene. Likewise, pSE143 plasmid with a umu C::lacZ fusion was introduced in S. typhimurium. Afterwards, induction of these SOS genes was studied. Results obtained show that the basal transcription of both recA and sfiA genes of E. coli was higher in S. typhimurium than in E. coli. Nevertheless, induction of recA and sfiA genes by UV-irradiation and mitomycin C was higher in E. coli than in S. typhimurium. On the other hand, umuC gene of E. coli presents the same basal level of transcription in both E. coli and S. typhimurium species, although induction of this gene by UV-irradiation and mitomycin C was higher in S. typhimurium than in E. coli. Therefore, the plasmid pUA25 constructed in this work may be used to introduce, using the Lambda phage as a vector, the SOS genes of E. coli in other bacterial species which may be useful to study the relationship between their respective SOS systems.


Effect of hydrocortisone on macrophage response to lymphokine.
To define the suppressive effects of corticosteroids on mononuclear phagocyte antiprotozoal activity, normal resident peritoneal macrophages were exposed to hydrocortisone (HC) before, during, and after in vitro activation with cell-free supernatants (lymphokines). The presence of pharmacological concentrations of HC before or during lymphokine activation prevented normal macrophages from acquiring the capacity to either respond oxidatively to Toxoplasma gondii ingestion or to inhibit intracellular toxoplasma replication. HC had no effect, however, on the cells fully stimulated by lymphokine or on macrophages previously activated in vivo. These findings indicate that although HC does not impair the ability of activated macrophages to control intracellular protozoan infection, it does compromise the antimicrobial activity of the cell-mediated immune system by rendering normal macrophages unresponsive to lymphokine.


Prenatal exposure to β2-adrenergic receptor agonists and risk of autism spectrum disorders.
This study aims to investigate the association between prenatal exposure to terbutaline and other β2 adrenergic receptor (B2AR) agonists and autism spectrum disorders (ASDs). The methodology used is a case-control study among children born from 1995 to 1999 at Kaiser Permanente Northern California hospitals. Cases (n = 291) were children with an ASD diagnosis; controls (n = 284) were children without ASDs, randomly sampled and frequency-matched to cases on sex, birth year, and delivery hospital. Exposure to B2AR agonists during 30 days prior to conception and each trimester of pregnancy was ascertained from prenatal medical records and health plan databases. The frequency of exposure to any B2AR agonist during pregnancy was similar for mothers of children with ASD and mothers of controls (18.9% vs. 14.8%, P = 0.19). Exposure to B2AR agonists other than terbutaline was not associated with an increased risk for ASDs. However, terbutaline exposure for >2 days during the third trimester was associated with more than a fourfold increased risk for ASDs independent of indication although the limited sample size resulted in an imprecise and nonsignificant effect estimate (OR(adj) = 4.4; 95% confidence interval, 0.8-24.6). This analysis does not offer evidence linking B2AR exposure in pregnancy with autism risk. However, exposure to terbutaline during the third trimester for >2 days may be associated with an increased risk of autism. Should this result be confirmed in larger samples, it would point to late pregnancy as an etiologic window of interest in autism risk factor research.


Preparation of purified suspensions of Coxiella burneti by Genetron extraction followed by continuous-flow ultracentrifugation.
A method for the preparation of purified suspensions of Coxiella burneti by Genetron extraction followed by continuous-flow density gradient ultracentrifugation is described. Both phases of the Henzerling strain of C. burneti were found in a zone between 53 and 65% (w/w) sucrose. Based on chemical assays, the Genetron zonal rickettsial suspensions were found to be as pure as the rickettsial suspensions which were prepared by the ether extraction method currently in use for producing Q fever vaccines for human use.


Influence of sulphuric acid contaminants on Fricke dosimetry.
The sulphuric acid used for the preparation of the Fricke dosimeter solution may contain trace impurities that can affect the yield of ferric ions. Two methods, pre-irradiation or oxidation with hydrogen peroxide, have been proposed to reduce the influence of these impurities. Fricke users sometimes omit this treatment. In the present work Fricke solutions prepared from six different brands and qualities of sulphuric acid were compared in order to study any influences of the acid on the ferric ion yield. It was shown that the use of analytical grade sulphuric acid from one manufacturer resulted in a reduction of the ferric ion yield of about 5% at an absorbed dose of approximately 20 Gy. If this solution were to be used for an absolute dose determination together with epsilon(m) G values from the literature the absorbed dose would be underestimated by the same amount.


Successful Use of BRAF/MEK Inhibitors as a Neoadjuvant Approach in the Definitive Treatment of Papillary Craniopharyngioma.
Craniopharyngiomas are rare tumors that arise in the suprasellar region of the brain and are known for their aggressive nature despite their WHO grade I. This is due to the complex neuroanatomy of the sellar/suprasellar region and their proximity to the optic nerve apparatus, hypothalamic-pituitary tract, and other critical neuroanatomical structures. Definitive treatment is based on a multidisciplinary approach and often involves a combination of surgical, radiation, and medical therapy. However, there is high morbidity associated with surgery and RT due to the complex neuroanatomy of this region. Recently, BRAFV600E somatic mutation has been identified in most papillary craniopharyngiomas. This discovery has led to the novel use of RAF pathway inhibitors to treat these tumors. We report the successful use of dabrafenib (BRAF inhibitor) and trametinib (mitogen-activated protein kinase kinase inhibitor) in the neoadjuvant setting followed by definitive stereotactic radiosurgery. We propose an algorithm based on available literature on the integration of targeted therapy in the management of papillary craniopharyngiomas. Our observations, together with prior case reports, advocate the incorporation of targeted therapy for unresectable craniopharyngiomas and reinforce that treatment with dual-targeted therapy is safe and effective.


Listeria monocytogenes in the Chinese food system: strain characterization through partial actA sequencing and tissue-culture pathogenicity assays.
Human listeriosis is generally caused by consumption of ready-to-eat (RTE) foods that are stored for extended periods of time at refrigeration temperatures and that permit the growth of the causative agent, Listeria monocytogenes. Food-consumption patterns in China are undergoing rapid changes and more regular consumption of refrigerated-storage RTE foods may increase the risk of human listeriosis. In total, 40 L. monocytogenes isolates were obtained from food (n=32) and sewage (n=6) samples and from two human listeriosis cases that occurred in China. All isolates were characterized into molecular subtypes by DNA sequencing of the 597 bp 3'-terminal region of the virulence gene actA. Sequence data were used to classify the 40 Chinese L. monocytogenes isolates into sequence types and phylogenetic lineages, and to compare the sequence types of the Chinese isolates with those of isolates from the USA. Phylogenetic analyses showed that the Chinese isolates could be separated into two genetic lineages, with 14 and 26 isolates belonging to lineages I and II, respectively. Lineage II could be subdivided further into two clusters, IIA and IIB. Lineages I and II were identical to the two lineages described previously among US L. monocytogenes isolates. In total, 14 actA sequence types could be differentiated among the 40 Chinese L. monocytogenes isolates; two specific actA sequence types were found among both Chinese and US isolates. Isolates belonging to lineage II showed a significantly lower ability to invade and multiply within human intestinal epithelial Caco-2 cells than lineage I isolates. It was concluded that DNA sequencing of the 3'-terminal region of actA appears to be an effective method for rapid subtype and lineage classification of L. monocytogenes. As strains belonging to lineages I and II have previously been found among isolates from Europe and North America, these results show that L. monocytogenes clonal groups found in China are very similar to those found in the USA. Many L. monocytogenes strains may thus represent globally distributed clonal types. Together with the first description of two human listeriosis cases in China, these data indicate that changes in food-distribution and -consumption patterns in China and other countries will probably lead to the emergence of human listeriosis as a food-safety issue, as virulent strains of this pathogen appear to be present in the Chinese food supply.


Validity and reliability of Persian version of Chronic Liver Disease Questionnaire (CLDQ).
The aim of this study is to test the psychometric properties of the Persian version of the Chronic Liver Disease Questionnaire (CLDQ) in Iranian candidates for liver transplantation. One hundred and fifty-five consecutive adult patients awaiting liver transplantation completed the Persian version of CLDQ and the short-form health survey (SF-36). The etiology of cirrhosis, Child Pugh classification and Model for End stage Liver Disease (MELD) scores were taken from medical records. The scaling success rate for convergent validity was 100% for all domains, and the success rate for item discriminant validity was 95.8% (139/145). The internal consistency (Cronbach α) for the domains ranged from 0.65 to 0.89. Multitrait-multimethod correlation matrix and factor analysis revealed that the CLDQ and SF-36 measure different constructs of quality of life. The Persian version of the CLDQ, a disease-specific questionnaire for measuring health-related quality of life, is accepted by liver transplantation candidates with adequate reliability and validity. There is no significant correlation of Child Pugh classification and MELD score with quality of life.