Day-to-day reliability of basal heart rate and short-term and ultra short-term heart rate variability assessment by the Equivital eq02+ LifeMonitor in US Army soldiers.

INTRODUCTION: The present study determined the (1) day-to-day reliability of basal heart rate (HR) and HR variability (HRV) measured by the Equivital eq02+ LifeMonitor and (2) agreement of ultra short-term HRV compared with short-term HRV. METHODS: Twenty-three active-duty US Army Soldiers (5 females, 18 males) completed two experimental visits separated by >48 hours with restrictions consistent with basal monitoring (eg, exercise, dietary), with measurements after supine rest at minutes 20-21 (ultra short-term) and minutes 20-25 (short-term). HRV was assessed as the SD of R-R intervals (SDNN) and the square root of the mean squared differences between consecutive R-R intervals (RMSSD). RESULTS: The day-to-day reliability (intraclass correlation coefficient (ICC)) using linear-mixed model approach was good for HR (0.849, 95% CI: 0.689 to 0.933) and RMSSD (ICC: 0.823, 95% CI: 0.623 to 0.920). SDNN had moderate day-to-day reliability with greater variation (ICC: 0.689, 95% CI: 0.428 to 0.858). The reliability of RMSSD was slightly improved when considering the effect of respiration (ICC: 0.821, 95% CI: 0.672 to 0.944). There was no bias for HR measured for 1 min versus 5 min (p=0.511). For 1 min measurements versus 5 min, there was a very modest mean bias of -4 ms for SDNN and -1 ms for RMSSD (p≤0.023). CONCLUSION: When preceded by a 20 min stabilisation period using restrictions consistent with basal monitoring and measuring respiration, military personnel can rely on the eq02+ for basal HR and RMSSD monitoring but should be more cautious using SDNN. These data also support using ultra short-term measurements when following these procedures.

'Super boots' for soldiers: theoretical ergogenic and thermoprotective benefits of energetically optimised military combat boots.

Soldiers typically perform physically demanding tasks while wearing military uniforms and tactical footwear. New research has revealed a substantial increase of ~10% in energetic cost of walking when wearing modern combat boots versus running shoes. One approach to mitigating these costs is to follow in the footsteps of recent innovations in athletic footwear that led to the development of 'super shoes', that is, running shoes designed to lower the energetic cost of locomotion and maximise performance. We modelled the theoretical effects of optimised combat boot construction on physical performance and heat strain with the intent of spurring similarly innovative research and development of 'super boots' for soldiers. We first assessed the theoretical benefits of super boots on 2-mile run performance in a typical US Army soldier using the model developed by Kipp and colleagues. We then used the Heat Strain Decision Aid thermoregulatory model to determine the metabolic savings required for a physiologically meaningful decrease in heat strain in various scenarios. Combat boots that impart a 10% improvement in running economy would result in 7.9%-15.1% improvement in 2-mile run time, for faster to slower runners, respectively. Our thermal modelling revealed that a 10% metabolic savings would more than suffice for a 0.25°C reduction in heat strain for the vast majority of work intensities and durations in both hot-dry and hot-humid environments. These findings highlight the impact that innovative military super boots would have on physical performance and heat strain in soldiers, which could potentially maximise the likelihood of mission success in real-world scenarios.

Clinical detection of Hepatitis C viral infection by yeast-secreted HCV-core:Gold-binding-peptide.

Access to affordable and field deployable diagnostics are key barriers to the control and eradication of many endemic and emerging infectious diseases. While cost, accuracy, and usability have all improved in recent years, there remains a pressing need for even less expensive and more scalable technologies. To that end, we explored new methods to inexpensively produce and couple protein-based biosensing molecules (affinity reagents) with scalable electrochemical sensors. Previous whole-cell constructs resulted in confounding measurements in clinical testing due to significant cross-reactivity when probing for host-immune (antibody) response to infection. To address this, we developed two complimentary strategies based on either the release of surface displayed or secretion of fusion proteins. These dual affinity biosensing elements couple antibody recognition (using antigen) and sensor surface adhesion (using gold-binding peptide-GBP) to allow single-step reagent production, purification, and biosensor assembly. As a proof-of-concept, we developed Hepatitis C virus (HCV)-core antigen-GBP fusion proteins. These constructs were first tested and optimized for consistent surface adhesion then the assembled immunosensors were tested for cross-reactivity and evaluated for performance in vitro. We observed loss of function of the released reagents while secreted constructs performed well in in vitro testing with 2 orders of dynamic range, and a limit of detection of 32 nM. Finally, we validated the secreted platform with clinical isolates (n = 3) with statistically significant differentiation of positive vs. non-infected serum (p < 0.0001) demonstrating the ability to clearly distinguish HCV positive and negative clinical samples.

Lithium-responsive genes and gene networks in bipolar disorder patient-derived lymphoblastoid cell lines.

Lithium (Li) is the mainstay mood stabilizer for the treatment of bipolar disorder (BD), although its mode of action is not yet fully understood nor is it effective in every patient. We sought to elucidate the mechanism of action of Li and to identify surrogate outcome markers that can be used to better understand its therapeutic effects in BD patients classified as good (responders) and poor responders (nonresponders) to Li treatment. To accomplish these goals, RNA-sequencing gene expression profiles of lymphoblastoid cell lines (LCLs) were compared between BD Li responders and nonresponders with healthy controls before and after treatment. Several Li-responsive gene coexpression networks were discovered indicating widespread effects of Li on diverse cellular signaling systems including apoptosis and defense response pathways, protein processing and response to endoplasmic reticulum stress. Individual gene markers were also identified, differing in response to Li between BD responders and nonresponders, involved in processes of cell cycle and nucleotide excision repair that may explain part of the heterogeneity in clinical response to treatment. Results further indicated a Li gene expression signature similar to that observed with clonidine treatment, an α2-adrenoceptor agonist. These findings provide a detailed mechanism of Li in LCLs and highlight putative surrogate outcome markers that may permit for advanced treatment decisions to be made and for facilitating recovery in BD patients.

Characterization of the complexity in short oscillating time series: An application to seismic airgun detonations.

Extracting frequency-derived parameters allows for the identification and characterization of acoustic events, such as those obtained in passive acoustic monitoring applications. Situations where it is difficult to achieve the desired frequency resolution to distinguish between similar events occur, for example, in short time oscillating events. One feasible approach to make discrimination among such events is by measuring the complexity or the presence of non-linearities in a time series. Available techniques include the delay vector variance (DVV) and recurrence plot (RP) analysis, which have been used independently for statistical testing, however, the similarities between these two techniques have so far been overlooked. This work suggests a method that combines the DVV method with the recurrence quantification analysis parameters of the RP graphs for the characterization of short oscillating events. In order to establish the confidence intervals, a variant of the pseudo-periodic surrogate algorithm is proposed. This allows one to eliminate the fine details that may indicate the presence of non-linear dynamics, without having to add a large amount of noise, while preserving more efficiently the phase-space shape. The algorithm is verified on both synthetic and real world time series.

Kaposi sarcoma-associated herpesvirus promotes tumorigenesis by modulating the Hippo pathway.

Kaposi sarcoma-associated herpesvirus (KSHV) is an oncogenic virus and the culprit behind the human disease Kaposi sarcoma (KS), an AIDS-defining malignancy. KSHV encodes a viral G-protein-coupled receptor (vGPCR) critical for the initiation and progression of KS. In this study, we identified that YAP/TAZ, two homologous oncoproteins inhibited by the Hippo tumor suppressor pathway, are activated in KSHV-infected cells in vitro, KS-like mouse tumors and clinical human KS specimens. The KSHV-encoded vGPCR acts through Gq/11 and G12/13 to inhibit the Hippo pathway kinases Lats1/2, promoting the activation of YAP/TAZ. Furthermore, depletion of YAP/TAZ blocks vGPCR-induced cell proliferation and tumorigenesis in a xenograft mouse model. The vGPCR-transformed cells are sensitive to pharmacologic inhibition of YAP. Our study establishes a pivotal role of the Hippo pathway in mediating the oncogenic activity of KSHV and development of KS, and also suggests a potential of using YAP inhibitors for KS intervention.

Ear-EEG from generic earpieces: a feasibility study.

The use of brain monitoring based on EEG, in natural environments and over long time periods, is hindered by the limited portability of current wearable systems, and the invasiveness of implanted systems. To that end, we introduce an ear-EEG recording device based on generic earpieces which meets key patient needs (discreet, unobstrusive, user-friendly, robust) and that is low-cost and suitable for off-the-shelf use; thus promising great advantages for healthcare applications. Its feasibility is validated in a comprehensive comparative study with our established prototype, based on a personalized earpiece, for a key EEG paradigm.

Auditory evoked responses from Ear-EEG recordings.

A method for brain monitoring based on measuring electroencephalographic (EEG) signals from electrodes placed in-the-ear (Ear-EEG) was recently proposed. The Ear-EEG recording methodology provides a non-invasive, discreet and unobtrusive way of measuring electrical brain signals and has great potential as an enabling method for brain monitoring in everyday life. This work aims at further establishing the Ear-EEG recording methodology by considering auditory evoked potentials, and by comparing Ear-EEG responses with conventional on-scalp recordings and with well established results from the literature. It is shown that both steady state and transient responses can be obtained from Ear-EEG, and that these responses have similar characteristics and quality compared to EEG obtained from conventional on-scalp recordings.

Estimating human response to taste using EEG.

In order to implement affective computing, there have been several studies to elicit human emotion using audio and video stimuli or by recalling previous events. Taste-elicited emotion has also been investigated using food to induce different levels of pleasure. This is monitored using a range of methods, from questionnaire feedback to electrophysiological responses of autonomic nervous system (ANS) and central nervous system (CNS). In this work, we establish that emotions elicited by taste can be monitored using electroencephalogram (EEG), and, for rigour, compare the response to a taste stimulus against the response to the recall of the same taste. The character of emotions were assessed using a subjective measurement, the hedonic score, which describes the pleasant or unpleasant moods of subjects in response to each taste. The classification performance of EEG responses shows excellent separability between the different emotions induced by different tastes. In addition, it is shown that emotion elicited by taste recall is stronger than the stimulus-elicited emotion.

An in-the-ear platform for recording electroencephalogram.

We introduce a novel approach to brain monitoring based on electroencephalogram (EEG) recordings from within the ear canal. While existing clinical and wearable systems are limited in terms of portability and ease of use, the proposed in-the-ear (ITE) recording platform promises a number of advantages including ease of implementation, minimally intrusive electrodes and enhanced accuracy (fixed electrode positions). It thus facilitates a crucial step towards the design of brain computer interfaces that integrate naturally with daily life. The feasibility of the ITE concept is demonstrated with recordings made from electrodes embedded on an earplug which are benchmarked against conventional scalp electrodes for a classic EEG paradigm.

A Yarbus-style experiment to determine auditory attention.

This paper presents an analysis of the merits of the original Yarbus experiment on eye movements with respect to judgments on differences in cognitive layer processes. The principles thus derived are applied to the development of an equivalent auditory experiment where, instead of eye movements, the response of the subject is observed by EEG measurements. Results from a preliminary trial are also included in which EEG analysis is used to ascertain the attended sound source in a multiple sound source environment. The investigation is part of ongoing research to improve the usefulness of hearing instruments and is also relevant in relation to other scientific investigations concerning the processing of sounds in complex acoustical environments by the human brain.

Differential effects of HIV-1 protease inhibitors on dendritic cell immunophenotype and function.

Recent findings show that human immunodeficiency virus (HIV)-1 protease inhibitors designed to specifically inhibit the aspartic protease of HIV-1 nonetheless exert various effects on immune cell function in vitro and in vivo. Dendritic cells (DC), central players of the immune system, express several aspartic proteases that are important for DC function. In the present study, we demonstrate that all of the HIV-1 protease inhibitors tested affect DC maturation. In addition, saquinavir had a strong inhibitory effect on the T-cell stimulatory capacity of mature DC. In contrast, indinavir had only a slight effect on DC induced T-cell proliferation and allowed efficient transduction of DC with a replication-incompetent HIV-1 vector designed for DC-based immunotherapy. HIV-1 protease inhibitors that have little or no effect on DC function may be preferable for combination with immunotherapy for HIV/AIDS.

Topical treatment of cutaneous lesions of acquired immunodeficiency syndrome-related Kaposi sarcoma using alitretinoin gel: results of phase 1 and 2 trials.

OBJECTIVE: To evaluate the efficacy and safety of topical alitretinoin gel (9-cis-retinoic acid [LGD1057], Panretin gel; Ligand Pharmaceuticals, Inc, San Diego, Calif) in cutaneous Kaposi sarcoma (KS). DESIGN: Open-label, within-patient, controlled, dose-escalating phase 1 and 2 clinical trials. In all patients, 1 or more cutaneous KS lesions were treated with alitretinoin gel, and at least 2 other lesions served as untreated controls for up to 16 weeks. Alitretinoin (0.05% or 0.1% gel) was applied twice daily for the first 2 weeks and up to 4 times daily thereafter, if tolerated. SETTING: Nine academic clinical centers. PATIENTS: One hundred fifteen patients with biopsy-proven acquired immunodeficiency syndrome (AIDS)-related KS. MAIN OUTCOME MEASURES: AIDS Clinical Trials Group response criteria. RESULTS: Statistically significant clinical responses were observed in 31 (27%) of 115 patients for the group of treated index lesions compared with 13 (11%) for the group of untreated control lesions (P<.001). Responses occurred with low CD4(+) lymphocyte counts (<200 cells/microL) and in some patients with refractory response to previous systemic anti-KS therapy. The incidence of disease progression was significantly lower for treated index lesions compared with untreated control lesions (39/115 [34%] vs 53/115 [46%]; P =.02). Alitretinoin gel generally was well tolerated, with 90% of treatment-related adverse events confined to the application site and only mild or moderate in severity. CONCLUSIONS: Alitretinoin gel has significant antitumor activity as a topical treatment for AIDS-related KS lesions, substantially reduces the incidence of disease progression in treated lesions, and is generally well tolerated.

Evolution of envelope sequences of human immunodeficiency virus type 1 in cellular reservoirs in the setting of potent antiviral therapy.

In human immunodeficiency virus (HIV)-infected patients treated with potent antiretroviral therapy, the persistence of latently infected cells may reflect the long decay half-life of this cellular reservoir or ongoing viral replication at low levels with continuous replenishment of the population or both. To address these possibilities, sequences encompassing the C2 and V3 domains of HIV-1 env were analyzed from virus present in baseline plasma and from viral isolates obtained after 2 years of suppressive therapy in six patients. The presence of sequence changes consistent with evolution was demonstrated for three subjects and correlated with less complete suppression of viral replication, as indicated by the rapidity of the initial virus load decline or the intermittent reappearance of even low levels of detectable viremia. Together, these results provide evidence for ongoing replication. In the remaining three patients, virus recovered after 2 years of therapy was either genotypically contemporary with or ancestral to virus present in plasma 2 years before, indicating that virus recovery had indeed resulted from activation of latently infected cells.

A disulfide-bound HIV-1 V3 loop sequence on the surface of human rhinovirus 14 induces neutralizing responses against HIV-1.

An immunogenic sequence from the V3 loop of the MN isolate of human immunodeficiency virus type 1 (HIV-1), His-Ile-Gly-Pro-Gly-Arg-Ala-Phe, was transplanted onto a surface loop of the VP2 capsid protein of human rhinovirus 14. To optimize for virus viability and immunogenicity of the transplanted sequence, the HIV sequence was flanked by (1) a cysteine residue that could form a disulfide bond and (2) randomized amino acids (in either of two arrangements) to generate numerous presentations of the Cys-Cys loop. The location for engineering in VP2 was chosen by searching the geometries of disulfide-bound loops in known protein structures. A model for the structure of the transplanted V3 loop sequence was developed using molecular dynamics and energy minimization calculations. Proteolytic digestion with and without reducing agent demonstrated the presence of the disulfide bond in the chimeric virus examined. Monoclonal and polyclonal antibodies directed against the V3 region of the HIV-1MN strain potently neutralized two chimeric viruses. Guinea pig antisera against two chimeric viruses were able to neutralize HIV-1MN and HIV-1ALA-1 in cell culture. The ability of chimeric viruses to elicit antibodies capable of neutralizing the source of the transplanted sequence could be favorable for vaccine development.

Mobilization of peripheral blood progenitor cells for human immunodeficiency virus-infected individuals.

Gene therapy is becoming one of the most promising modalities for the treatment of acquired immunodeficiency syndrome. The purpose of this study was to investigate the mobilization and collection of peripheral blood progenitor cells from human immunodeficiency virus (HIV)-infected individuals using granulocyte colony-stimulating factor (G-CSF). A total of 10 patients (9 male, 1 female; median age 36.5 years) with varying circulating CD4+ cell counts (13.9-1467/microL) were administered 10 microg/kg G-CSF daily for 6 days. Peripheral white blood cells (WBCs), CD34+ cell counts, lymphocyte subsets, and plasma viremia were monitored before each G-CSF injection. An average sixfold increase in WBCs was observed, which stabilized on day 4 or thereafter. The level of CD34+ cells was increased by 20-fold, and did not differ between days 5 and 6. Smaller increases in CD4+, CD8+, and CD4+CD8+ cells were observed. HIV viral load, as measured by RNA copy number in plasma, was not significantly altered by G-CSF administration. The leukapheresis product (LP), collected on day 7, contained an average of 6.25+/-4.52 (mean +/- standard deviation) x 10(10) WBCs and 3.08+/-2.98 x 10(6) CD34+ cells/kg. The levels of different CD34+ cell subsets were similar to those in the LPs of G-CSF-mobilized healthy individuals from an earlier study. Primitive hematopoietic cells (CD38- and CD38-HLA-DR+ cells) were detected in LPs (1.19+/-0.46% and 0.87+/-0.23%, respectively, of CD34+ cells). All parameters (WBC counts, lymphocyte populations, CD34+ cells, and HIV-1 RNA copies) measured 3 weeks after leukapheresis returned to baseline values. The administration of G-CSF was well tolerated by the HIV patients; side effects included bone pain, headache, flulike symptoms, and fatigue. There were no correlations between baseline CD4+ cell count and the WBCs, mononuclear cells, or CD34+ cells collected in the LP. Similarly, no correlation existed between baseline CD4+ and CD34+ cells, peak CD34+ cells, or days to achieve peak CD34+ cell counts after G-CSF mobilization. Our results showed that: (1) maximal mobilization can be achieved after 4 days of G-CSF administration; (2) therapeutic quantities of hematopoietic cells can be collected and used for gene therapy; and (3) G-CSF administration is well tolerated and does not cause a clinically significant increase in viremia.

A controlled, Phase 1 clinical trial to evaluate the safety and effects in HIV-1 infected humans of autologous lymphocytes transduced with a ribozyme that cleaves HIV-1 RNA.

This Phase I study, "Ribozyme Gene Therapy of HIV-1 Infection" (UCSD HSC #971072, FDA BB-IND 6405), is a prospective, open-label trial of infusion of autologous gene-altered cells into asymptomatic HIV-1 seropositive individuals. The objectives of this trial are to test the safety, feasibility, and potential efficacy of T-cell ribozyme gene therapy of HIV-1 infection. To accomplish this, autologous CD8-depleted mononuclear cells are transduced with ribozyme expressing or control murine retroviral vectors, expanded ex vivo, and and infused. Subjects are monitored intensively to determine effects of infusion on HIV burden and replication. In addition, in vivo survival of control and ribozyme transduced cells is followed in an effort to obtain evidence of proof of concept. A unique strategy of sample blinding is introduced in this protocol, wherein both subject and control specimens are supplied to the research laboratory as coded samples, spiking blood from HIV seropositive volunteers matched for CD4 lymphocyte count with known but varying numbers of cells transduced with each vector. While this study is still in progress, preliminary results indicate that infusion of gene-altered, activated T-cells in HIV infected patients is safe, and that transduced cells can persist for long intervals in HIV-infected subjects. Results also suggest ribozyme transduced cells may possess a survival advantage in vivo.

Genetic variation in a human immunodeficiency virus type 2 live-virus Macaca nemestrina vaccine model.

Four pigtailed macaques were inoculated with an infectious, apathogenic human immunodeficiency virus type 2 (HIV-2) molecular clone (HIV-2KR) and subsequently challenged with a highly pathogenic strain, HIV-2287, together with two naive control animals. After challenge, two animals inoculated with a high dose of the immunizing strain were protected from CD4 decline and immunodeficiency. To examine the role of genetic heterogeneity in protection, fragments of the env gene were amplified from peripheral blood mononuclear cell DNA and plasma RNA of challenged animals by PCR, examined by using a heteroduplex tracking assay (HTA), and sequenced. By HTA, variation was detected principally within the V1 and V2 regions of envelope. Extent of variation in viral DNA clones as assessed by HTA correlated with inoculum size, as did the degree of variation in sequences of clones derived from viral DNA. Conversely, a rapid reduction in the number of plasma viral RNA variants was noted by HTA at 8 weeks postinfection in protected animals; this reduction was not present in naive or unprotected macaques. Sequences derived from plasma viral RNA were found to be more closely related than corresponding viral DNA sequences, and protection correlated with a significant reduction in variation in plasma RNA sequences in animals given the identical inocula of HIV-2287. Nonsynonymous mutations were significantly less prevalent in the protected animals. An additional potential glycosylation site was predicted to be present in the V2 region in all but one clone, and amino acid signatures related to protection were identified in viral DNA and RNA clones within both the V1 and V2 regions. Examination of the role of viral variation in this HIV-2 live-virus vaccine model may provide valuable insights into immunopathogenesis.

CXCR4 is required by a nonprimate lentivirus: heterologous expression of feline immunodeficiency virus in human, rodent, and feline cells.

A heterologous feline immunodeficiency virus (FIV) expression system permitted high-level expression of FIV proteins and efficient production of infectious FIV in human cells. These results identify the FIV U3 element as the sole restriction to the productive phase of replication in nonfeline cells. Heterologous FIV expression in a variety of human cell lines resulted in profuse syncytial lysis that was FIV env specific, CD4 independent, and restricted to cells that express CXCR4, the coreceptor for T-cell-line-adapted strains of human immunodeficiency virus. Stable expression of human CXCR4 in CXCR4-negative human and rodent cell lines resulted in extensive FIV Env-mediated, CXCR4-dependent cell fusion and infection. In feline cells, stable overexpression of human CXCR4 resulted in increased FIV infectivity and marked syncytium formation during FIV replication or after infection with FIV Env-expressing vectors. The use of CXCR4 is a fundamental feature of lentivirus biology independent of CD4 and a shared cellular link to infection and cytopathicity for distantly related lentiviruses that cause AIDS. Their conserved use implicates chemokine receptors as primordial lentivirus receptors.