Nanosecond X-Ray Photon Correlation Spectroscopy on Magnetic Skyrmions.

We report an x-ray photon correlation spectroscopy method that exploits the recent development of the two-pulse mode at the Linac Coherent Light Source. By using coherent resonant x-ray magnetic scattering, we studied spontaneous fluctuations on nanosecond time scales in thin films of multilayered Fe/Gd that exhibit ordered stripe and Skyrmion lattice phases. The correlation time of the fluctuations was found to differ between the Skyrmion phase and near the stripe-Skyrmion boundary. This technique will enable a significant new area of research on the study of equilibrium fluctuations in condensed matter.

Observation of deflection of a beam of multi-GeV electrons by a thin crystal.

We report on an experiment performing channeling and volume reflection of a high-energy electron beam using a quasimosaic, bent silicon (111) crystal at the End Station A Test Beam at SLAC. The experiment uses beams of 3.35 and 6.3 GeV. In the channeling orientation, deflections of the beam of 400 µrad for both energies with about 22% efficiency are observed, while in the volume-reflection orientation, deflection of the beam by 120 µrad at 3.35 GeV and by 80 µrad at 6.3 GeV is observed with 86%-95% efficiency. Quantitative measurements of the channeling efficiency, surface transmission, and dechanneling length are taken. These are the first quantitative measurements of channeling and volume reflection using a primary beam of multi-GeV electrons.

Highly efficient star formation in NGC 5253 possibly from stream-fed accretion.

Gas clouds in present-day galaxies are inefficient at forming stars. Low star-formation efficiency is a critical parameter in galaxy evolution: it is why stars are still forming nearly 14 billion years after the Big Bang and why star clusters generally do not survive their births, instead dispersing to form galactic disks or bulges. Yet the existence of ancient massive bound star clusters (globular clusters) in the Milky Way suggests that efficiencies were higher when they formed ten billion years ago. A local dwarf galaxy, NGC 5253, has a young star cluster that provides an example of highly efficient star formation. Here we report the detection of the J = 3→2 rotational transition of CO at the location of the massive cluster. The gas cloud is hot, dense, quiescent and extremely dusty. Its gas-to-dust ratio is lower than the Galactic value, which we attribute to dust enrichment by the embedded star cluster. Its star-formation efficiency exceeds 50 per cent, tenfold that of clouds in the Milky Way. We suggest that high efficiency results from the force-feeding of star formation by a streamer of gas falling into the galaxy.

Demonstration of single-crystal self-seeded two-color x-ray free-electron lasers.

A scheme for generating two simultaneous hard-x-ray free-electron laser pulses with a controllable difference in photon energy is described and then demonstrated using the self-seeding setup at the Linac Coherent Light Source (LCLS). The scheme takes advantage of the existing LCLS equipment, which allows two independent rotations of the self-seeding diamond crystal. The two degrees of freedom are used to select two nearby crystal reflections, causing two wavelengths to be present in the forward transmitted seeding x-ray pulse. The free-electron laser system must support amplification at both desired wavelengths.

Effects of oxytocin, lipopolysaccharide (LPS), and polyunsaturated fatty acids on prostaglandin secretion and gene expression in equine endometrial explant cultures.

Increased secretion of prostaglandin F(2)α (PGF(2)α) within the uterus because of uterine inflammation can cause luteolysis and result in early embryonic loss. Supplementation with polyunsaturated fatty acids (PUFAs) has been shown to influence PG production in many species, although the effects on the mare remain unknown. The present study aimed to determine fatty acid uptake in equine endometrial explants and evaluate their influence on PG secretion and expression of enzymes involved in PG synthesis in vitro. Equine endometrial explants were treated with 100 µM arachidonic acid, eicosapentaenoic acid, or docosahexaenoic acid and then challenged with oxytocin (250 nM) or lipopolysaccharide (LPS; 1 µg/mL). Production of PGF(2)α and PG E(2) (PGE(2)) was measured, and mRNA expression of enzymes involved in PG synthesis was determined with quantitative real-time PCR. Media concentrations of PGF(2)α and PGE(2) were higher (P < 0.0001) from endometrial explants challenged with oxytocin or LPS compared with controls despite which fatty acid was added. Only DHA lowered (P < 0.0001) media concentrations of PGF(2)α and PGE(2) from explants. Endometrial explants stimulated with oxytocin had increased expression of PG-endoperoxide synthase 1 (PTGS1; P < 0.02), PG-endoperoxide synthase 2 (PTGS2; P < 0.001), PG F(2)α synthase (PGFS; P < 0.01), PG E(2) synthase (PGES; P < 0.01), and phospholipase A(2) (PLA(2); P < 0.005) compared with controls and regardless of fatty acid treatment; whereas stimulation with LPS increased expression of PTGS2 (P < 0.004), PGFS (P < 0.03), PGES (P < 0.01), and PLA(2) (P < 0.01) compared with controls and regardless of fatty acid treatment. Treatment with PUFAs, specifically DHA, can influence PG secretion in vitro through mechanisms other than enzyme expression.

Effects of dietary protein and bacterial lipopolysaccharide infusion on nitrogen metabolism and hormonal responses of growing beef steers.

Purified lipopolysaccharide (LPS) infusion in cattle induces clinical and metabolic responses similar to gram-negative bacterial infection. Effects of LPS and dietary protein on rectal temperature, serum hormones, haptoglobin, plasma urea N and AA, and N balance were evaluated in 24 steers (250 +/- 2.8 kg of BW). Treatments were a 2 x 3 factorial of LPS (0 vs. 1.5 microg/kg of BW; -LPS vs. +LPS) and diets containing (DM basis) 1) 14.5% CP, 11.6% ruminally degradable protein (RDP), and 2.9% ruminally undegradable protein (RUP; CP14.5CON); 2) 16.3% CP, 13.4% RDP, and 2.9% RUP (CP16RDP); and 3) 16.1% CP, 11.2% RDP, and 4.9% RUP (CP16RUP). Diet RDP and RUP were altered using casein, fish meal, and corn gluten meal. Steers were adapted to diets (1.1 Mcal/kg of NE(g); DM fed at 1.8% BW) for 14 d and were infused (intravenously 1 mL/min) with LPS (in 100 mL of saline) on d 15. Rectal temperature and serum cortisol, prolactin, haptoglobin, and insulin increased, glucose initially increased and then declined, and serum thyroxine and triiodothyronine decreased for +LPS vs. -LPS steers (LPS x hour; P < 0.01). Serum IGF-I was less (P < 0.01) for +LPS vs. -LPS steers. Plasma urea N increased in response to LPS (LPS x hour; P = 0.02) and was greater for +LPS steers fed CP16RDP and CP16RUP vs. CP14.5CON, but greater in -LPS steers fed CP16RUP vs. CP16RDP and CP14.5CON (LPS x diet; P = 0.04). Plasma Met, Thr, Leu, Ile, Phe, Trp, Gly, Ser, Asn, and Tyr decreased, and plasma Ala increased in response to LPS (LPS x hour; P < 0.01). Plasma Orn initially increased and then decreased in +LPS vs. -LPS steers (LPS x hour; P < 0.01). No LPS x diet interactions (P > or = 0.15) occurred for DM, OM, NDF and N intake, fecal excretion, or apparent digestibility. Dietary DM, OM, NDF, and N intake, and retained N were less (P < 0.01) for +LPS than -LPS steers. Total N intake, apparent N digestibility, and retained N were greater (P < or = 0.05) for steers fed CP16RDP and CP16RUP vs. CP14.5CON. An LPS x diet interaction (P = 0.05) occurred for N retention (% N intake) because N retention was less for +LPS than -LPS steers when fed CP14.5CON, but not different between +LPS and -LPS steers when fed CP16RDP and CP16RUP. These results demonstrate that LPS infusion alters serum hormones, plasma AA, and N balance in cattle and imply that growing steers exposed to LPS may require greater dietary protein concentrations to account for altered intake and metabolic AA demand.

Characterisation of a type 2 bovine viral diarrhoea virus isolated from cattle in the UK.

Two genotypes of bovine viral diarrhoea virus (BVDV) are recognised. Type 2 was first recognised when virulent strains caused significant losses among cattle in North America. Subsequently, BVDV type 2 has been found in many other countries, but recent studies have shown that only type 1 BVDV is circulating in the UK herds (sheep and cattle) with type 1a predominating. During routine genotyping of UK BVDV isolates, a type 2 isolate was identified. Phylogenetic analysis of the 5'-untranslated region of the viral genome showed it to be a BVDV type 2a, most similar to a low virulent US strain of BVDV type 2. Antigenic typing with a panel of monoclonal antibodies verified this classification. This is the first confirmed isolation of BVDV type 2 found circulating in the UK.

Changes in circulating insulin-like growth factor-I, insulin-like growth factor binding proteins, and leptin in weaned pigs infected with Salmonella enterica serovar Typhimurium.

One of the hallmarks of the pathophysiology of enteric disease in young pigs is reduced growth performance. This reduction in growth is associated with changes in the endocrine somatotropic growth axis. Our laboratory previously demonstrated that circulating insulin-like growth factor-I (IGF-I) was reduced in pigs infected with Salmonella enterica serovar Typhimurium (S. typhimurium) while circulating growth hormone remained unchanged. The objective of the current study was to determine if infection with S. typhimurium also was associated with changes in circulating IGF binding proteins (IGFBP). In addition, pigs experiencing active enteric disease have reduced feed intake. Because this inappetence may be related to systemic appetite reduction signals, we also evaluated circulating leptin in pigs undergoing active S. typhimurium-induced enteric disease. Crossbred pigs were penned in environmentally controlled rooms with free access to feed and water. Following an acclimation period, pigs were gavaged with 10(10) cfu of S. typhimurium (SAL; n=6) or were given a similar volume of sterile growth media (CON; n=6). Rectal temperatures and feed intakes were measured daily through 168 h to track the time course of the response to S. typhimurium infection. Samples of serum were obtained by jugular venipuncture at 0, 24, 48, 96 and 168 h after infection. Sera were frozen until evaluation for IGF-I by immunoradiometric assay (IRMA). In addition, sera were subjected to western ligand blotting utilizing 125I-IGF-I and 125I-IGF-II. Images were evaluated for total IGFBP and IGFBP-3 by densitometric analyses. Rectal temperature was increased in SAL pigs 24h post-infection (P<0.001) but not at other times. Feed intake was reduced in SAL pigs during the intervals 24-72 h (P<0.001) and 96-144 h (P<0.05) after infection. Serum IGF-I, expressed as a percentage of the 0 h concentration, was reduced in SAL pigs versus CON pigs at 48 h (28.1+/-18.7% versus 102.2+/-17.1%; P<0.01) and 96 h (20.0+/-18.7% versus 128.4+/-17.0%; P<0.0001) post-infection. Both total IGFBP and IGFBP-3, as estimated by ligand blotting, also were reduced in infected pigs at 48 h postchallenge (P<0.05). IGFBPs were similar between the two treatments at other sampling times. Concentrations of IGFBP-3 also were estimated utilizing an IRMA for human IGFBP-3. Serum IGFBP-3 was reduced in S. typhimurium-infected pigs at 24 h (P<0.01), 48 h (P<0.001), 96 h (P<0.001), and 168 h (P<0.05). Serum leptin levels were similar between SAL and CON pigs. The data suggest that swine enteric disease is associated with reduced circulating IGF-I and reductions in total IGFBP and IGFBP-3. However, serum leptin was not affected by enteric disease challenge.

An extragalactic supernebula confined by gravity.

Little is known about the origins of globular clusters, which contain hundreds of thousands of stars in a volume only a few light years across. Radiation pressure and winds from luminous young stars should disperse the star-forming gas and disrupt the formation of the cluster. Globular clusters in our Galaxy cannot provide answers; they are billions of years old. Here we report the measurement of infrared hydrogen recombination lines from a young, forming super star cluster in the dwarf galaxy NGC5253. The lines arise in gas heated by a cluster of about one million stars, including 4,000-6,000 massive, hot 'O' stars. It is so young that it is still enshrouded in gas and dust, hidden from optical view. The gases within the cluster seem bound by gravity, which may explain why the windy and luminous O stars have not yet blown away those gases. Young clusters in 'starbursting' galaxies in the local and distant Universe may also be gravitationally confined and cloaked from view.

Deconvolution of surface topology for quantification of initial wear in highly cross-linked acetabular components for THA.

Evaluation of the surface morphology of short-term retrieved cross-linked acetabular components requires differentiation between the features generated during machining and the smaller-scale morphologies generated during the in vivo wear process. Previously, the distinction between the waviness of machining and the roughness of wear has been related to the grain size of the UHMWPE. Here a low-frequency cutoff is proposed, based on the maximum spectral frequency of machining marks, rather than on the grain size of the bulk UHMWPE material, as a reliable method for deconvolving machining marks from in vivo wear following short-term implantation. To this end, as-machined articulating surfaces of conventional (GUR 1050) and two groups of highly cross-linked UHMWPE acetabular components were examined to determine whether they exhibited a periodic surface morphology with a well-defined spatial frequency. The surface frequency spectra revealed low-frequency peaks associated with the machining marks, which were unique to each type of implant. Furthermore, the surface frequency spectra appeared uniform within a single group of implants. Statistically significant differences in the surface roughness and waviness were observed between the three groups of new implants. Our research suggests that machining marks can be effectively deconvolved from the articulating surface with the use of a Fourier transform algorithm with a single cutoff frequency of 0.08 1/microm, corresponding to a wavelength of 12.5 microm. The results of this study provide a unified conceptual framework for discriminating between waviness and roughness of the articulating surface for machined orthopedic components. The distinction between waviness and roughness is expected to be crucial for the comprehensive evaluation of wear surfaces after short-term implantation, when machining marks may be partially worn away or plastically deformed in vivo.

Effects of a Quillaja saponaria extract on growth performance and immune function of weanling pigs challenged with Salmonella typhimurium.

Ninety-six pigs (initially 8.9 kg and 24 d of age) were used in a 28-d experiment to determine the effects of Quillaja saponaria extract (QS) on weanling pig growth performance and immune function in response to enteric disease challenge with Salmonella typhimurium (ST). Experimental treatments were arranged in a 2 x 4 factorial with main effects of disease challenge (control vs ST-challenge) and dietary addition of QS (0, 125, 250, or 500 mg/kg). Pigs were fed QS diets for 14 d and then challenged orally with ST or sterile media. There were no differences in ADG or ADFI among dietary treatments, but gain/feed ratio (G/ F) was depressed (P < 0.06) in pigs fed 250 mg/kg QS. ST-challenge reduced ADG (P < 0.05), ADFI (P < 0.05), and G/F (P < 0.05) 1 wk after challenge. Daily estimates revealed reductions in feed intake in ST-infected pigs on d 2 to 5 following infection (P < 0.05), and rectal temperature was increased maximally 2 d following infection (P < 0.05). There was a marked decline in serum IGF-I during the 6 d after ST-infection (P < 0.05). ST-challenge produced a rise (P < 0.05) in serum haptoglobin on d 7 after challenge, and serum alpha1-acid glycoprotein (AGP) in ST-challenged pigs also was elevated (P < 0.05) above controls on d 7 and 14 after challenge. Serum immunoglobulin (Ig) M increased (P < 0.05) over time in both groups, and serum IgM of ST-challenged pigs was greater than controls on d 7 after challenge (P < 0.05). Serum IgG was not affected by enteric disease challenge; however, on d 7 and 14 after disease challenge, serum IgG for both groups was greater (P < 0.05) than on d 0. Dietary QS had no significant influence on any of the end points used to characterize the acute phase response to ST-challenge. Phagocytic cell function was depressed in pigs fed 250 (P < 0.05) and 500 (P < 0.05) mg/kg as compared to pigs fed 125 mg/kg QS. Yet, there was no difference in phagocytic function among pigs fed 0, 250, or 500 mg/kg QS. We conclude that this model of enteric disease invokes an acute phase response accompanied by decreases in feed intake and serum IGF-I. Furthermore, dietary QS, at the levels fed in this study, appears to offer little benefit to growth performance or immune function in the presence or absence of ST-challenge.

Effects of Ascophyllum nodosum extract on growth performance and immune function of young pigs challenged with Salmonella typhimurium.

Ninety-five pigs (initially 7.1 kg and 24 d of age) were used in a 28-d experiment to determine the effects of Ascophyllum nodosum seaweed extract (ANOD) on young pig growth performance and immune function in response to enteric disease challenge with Salmonella typhimurium (ST). Experimental treatments were arranged in a 2 x 4 factorial with main effects of disease challenge (control vs ST-challenge) and dietary addition of ANOD (0, 0.5, 1.0, and 2.0% of diet). Pigs were fed ANOD diets for 14 d and then challenged orally with ST or sterile media. There were no main effects of ANOD on growth performance end points, although there were significant quadratic effects of ANOD on ADG (P < 0.04) and final weight (P < 0.003), both being greatest at 1.0% ANOD. There was a positive linear effect of ANOD inclusion on ADFI (P < 0.07) and a negative linear effect on the gain-to-feed ratio (G/F) (P < 0.05). ST-challenge reduced ADG (P < 0.05), ADFI (P < 0.05), and G/F (P < 0.05) in the first week following challenge. Daily estimates revealed reductions in feed intake in ST-infected pigs on d 2 to 4 following infection (P < 0.05). Rectal temperature was increased maximally 2 d following ST-infection (P < 0.05). A disease challenge x time interaction (P < 0.001) was observed for serum haptoglobin and alpha1-acid glycoprotein. Serum immunoglobulin M (IgM) was not influenced by disease challenge, but IgM declined (P < 0.001) in all pigs over time. Serum immunoglobulin G (IgG) also was not influenced by disease challenge, but IgG tended (P < 0.08) to increase over time. In vitro culture of porcine alveolar macrophages with 10 mg/mL ANOD elevated (P < 0.05) prostaglandin E2 (PGE2) production over that of controls at 3 and 24 h of culture. There was no interleukin-10 response by porcine splenocytes cultured in vitro with 0.005, 0.05, 0.5, or 5 mg/mL ANOD. We conclude that this model of enteric disease elicits an acute phase response that is accompanied by increased rectal temperature and diminished feed intake. Furthermore, our results indicate some beneficial effects of dietary ANOD on growth performance and no influence of dietary ANOD on immune response in the presence or absence of ST-challenge. However, high ANOD concentrations are capable of activating porcine alveolar macrophages in vitro to secrete PGE2.

Circulating cortisol, tumor necrosis factor-alpha interleukin-1beta, and interferon-gamma in pigs infected with Actinobacillus pleuropneumoniae.

This study evaluated the time course of systemic cytokine concentrations in an acute model of pneumonia in pigs challenged intranasally with Actinobacillus pleuropneumoniae. Feed intake and serum cortisol were measured as overt clinical and systemic markers of disease onset, respectively, and serum tumor necrosis factor-alpha, interleukin-1beta, and interferon-gamma as representative systemic inflammatory markers. Crossbred barrows (n = 15), approximately 5 wk of age, were used in the study. Pigs were housed in an environmentally controlled facility at 25 degrees C and under continuous illumination in pens measuring approximately 1.5 m2. Pigs had free access to water and an unmedicated diet. Approximately 1 wk prior to disease challenge, pigs were fitted nonsurgically with venous catheters. At challenge, pigs were given 5 x 10(8) CFU Actinobacillus pleuropneumoniae intranasally (n = 8) or a similar volume of sterile growth media intranasally (Control; n = 7). Feed intake was estimated by the change in feeder weight at 12-h intervals from -12 to 72 h relative to the time of disease challenge. Blood sampling began 12 h prior to challenge and continued until 72 h after challenge. Pigs were sampled at -12, -6, and 0 h, then at 90-min intervals until 12-h post-challenge, continuing at 3-h intervals until 24-h post-challenge, then again at 6-h intervals until 72 h after challenge. Serum was harvested and frozen until assayed for cortisol, tumor necrosis factor-alpha, interleukin-1beta, and interferon-gamma. Feed intake was reduced in Actinobacillus pleuropneumoniae pigs during the intervals 0 to 12 h (P < 0.001), 24 to 36 h (P < 0.001), 48 to 60 h (P <0.05), and 60 to 72 h (P < 0.05). TheActnobacillus pleuropneumoniae-challenged pigs had elevated serum cortisol from 180-min to 18-h post-challenge (P < 0.001) and also at 36 (P < 0.05), 42 (P < 0.001), and 60 (P < 0.05) h following infection. Circulating cytokines were not affected by disease challenge. Thus, in this experimental model of pneumonia, weaned pigs demonstrated expected behavioral and endocrine characteristics of disease in the absence of significant changes in circulating inflammatory cytokines.

The effects of an HIV-1 immunogen (Remune) on viral load, CD4 cell counts and HIV-specific immunity in a double-blind, randomized, adjuvant-controlled subset study in HIV infected subjects regardless of concomitant antiviral drugs.

OBJECTIVE: We examined the activity of an HIV-1 immunogen (Remune) on viral load, CD4 cells and HIV-1 specific immunity. METHODS: Plasma and peripheral blood mononuclear cells were obtained in a predefined random subset of subjects (n = 252) from a multicentre, double-blind, adjuvant-controlled phase III clinical endpoint study. RESULTS: The subjects treated with the HIV-1 immunogen had a significantly greater decline in viral load at multiple time points (P < 0.05), a trend towards increased CD4+ T cell counts and significantly enhanced HIV-1 specific immune responses as measured by HIV-1 lymphocyte proliferation (P < 0.001) compared to the adjuvant control group. Furthermore, in the HIV-1 immunogen treated group, enhanced HIV-1 specific lymphocyte proliferative immune responses were associated with decreased HIV-1 plasma RNA. CONCLUSION: These results suggest that, in a predefined, random subset of subjects, a beneficial effect of the HIV-1 immunogen was observed on viral load, CD4+ T cells, and HIV-specific immunity. These differences were observed in a background of multiple drug therapies. Ongoing trials are evaluating the effect of the combination of this HIV-1 specific, immune-based therapy with potent antiviral drug therapy on virological outcomes.

Tumor necrosis factor alpha and human immunodeficiency virus-specific functional immune responses after immunization with Gp120-depleted, inactivated HIV-1 in incomplete Freund's adjuvant (REMUNE) in HIV-1-seropositive subjects.

OBJECTIVE: We examined the relation between tumor necrosis factor-alpha (TNF-alpha) levels and human immunodeficiency virus type 1 (HIV-1)-specific functional immune responses, as measured by HIV-1 antigen-stimulated lymphocyte proliferation and beta-chemokine production after immunization with gp120-depleted, inactivated HIV-1 in incomplete Freund's adjuvant (i.e., HIV-1 Immunogen; REMUNE, The Immune Response Corporation, Carlsbad, CA, U.S.A.). STUDY DESIGN/METHODS: HIV-1-seropositive subjects who enrolled in an open-label study were immunized with REMUNE every 12 weeks and monitored for 60 weeks. HIV-1 antigen-stimulated lymphocyte proliferation and RANTES production were measured in peripheral blood mononuclear cells (PBMCs). TNF-alpha levels were measured in serum. RESULTS: TNF-alpha (P = 0.0003) significantly decreased and HIV-1 antigen-stimulated RANTES production (P = 0.002) and lymphocyte proliferation (P = 0.07) increased after immunization with REMUNE. TNF-alpha levels negatively correlated with HIV-1 antigen-stimulated RANTES production (r = -0.71; P = 0.0002) and lymphocyte proliferation (r = -0.37; P = 0.09). CONCLUSIONS: This study demonstrated decreased TNF-alpha levels with a concomitant augmentation of HIV-specific functional immunity in subjects immunized with REMUNE. Because TNF-alpha has been implicated in the induction of anergy in HIV-1 infection, the ability to decrease TNF-alpha may allow the immune system to respond to HIV and non-HIV antigens. Larger studies are being conducted to confirm the clinical utility of REMUNE in combination with potent antiviral drugs.

Relationship between membrane potential, delayed rectifier K+ currents and hypoxia in rat pulmonary arterial myocytes.

Pulmonary arteries constrict in response to hypoxia, a process thought to involve oxygen sensing by K+ channels. We therefore investigated the effects of hypoxia on voltage-activated K+ currents in myocytes isolated from rat small pulmonary arteries using the patch-clamp recording technique. Experiments with iberiotoxin and intracellularly applied Ca2+ chelating agents revealed that hypoxia (PO2, 20-30 mmHg; throughout) inhibited the Ca(2+)-insensitive component of the delayed voltage-activated outward K+ current. Hypoxia did not affect the membrane potential of these cells until they were depolarized by extracellular application of 20 mM K+, current injection or endothelin-1. Hypoxia caused little depolarization in the presence of prostaglandin F2 alpha, an agonist which was ineffective at inducing depolarization. These results suggest that an initial 'priming' depolarization may confer a sensitivity to hypoxia by activating delayed rectifier (Kv) channels. Once active, these channels can then be closed by hypoxia, leading to further depolarization. It is unlikely, therefore, that Kv channels are involved in controlling the resting membrane potential of these cells.

Effect of immunization with an inactivated gp120-depleted HIV-1 immunogen on beta-chemokine and cytokine production in subjects with HIV-1 infection.

OBJECTIVE: To measure beta-chemokine and cytokine production in HIV-1-infected subjects undergoing treatment with HIV-1 immunogen (REMUNE). DESIGN: Open label treatment study. METHODS: beta-Chemokine and cytokine production in peripheral blood mononuclear cell (PBMC) culture. RESULTS: Interferon-gamma production (p = 0.04) and lymphocyte proliferation (p = 0.001) to HIV-1 antigen-stimulated PBMCs increased after immunization with the HIV-1 immunogen. A correlation was demonstrated after immunization between HIV-1 antigen-stimulated lymphocyte proliferation and interferon-gamma levels (r = 0.53, p = 0.04). No significant change after immunization was seen for interleukin-4 production. A significant increase in mean levels of HIV-1 antigen-stimulated RANTES (i.e., regulated upon, activation normal T-cell expressed and secreted), was evident 1 month after immunization (p = 0.002) and remained elevated 3 months after immunization. RANTES production was decreased in CD8-depleted PBMC cultures. Mean serum HIV-1 RNA copy numbers and CD4 cell counts remained stable after immunization (p > 0.5). A correlation was demonstrated between HIV-1 antigen-stimulated interferon-gamma and RANTES production (r = 0.54, p = 0.002). CONCLUSIONS: This report describes an augmentation of beta-chemokines and TH1-type cytokines from PBMCs after immunization with the HIV-1 immunogen.

Cross-clade immune responses after immunization with a whole-killed gp120-depleted human immunodeficiency virus type-1 immunogen in incomplete Freund's adjuvant (HIV-1 immunogen, REMUNE) in human immunodeficiency virus type-1 seropositive subjects.

Lymphocyte proliferation responses to gp120-depleted HZ321 virus (clade A) antigen were compared to BAL human immunodeficiency virus (HIV) virus antigen (clade B) responses, clade E HIV virus antigen responses, and purified native p24 antigen responses in 15 human immunodeficiency virus type-1 (HIV-1) seropositive subjects immunized with a whole-killed inactivated gp120-depleted HIV-1 antigen in Incomplete Freund's adjuvant (HIV-1 immunogen, REMUNE). A significant increase in lymphocyte proliferation to HZ321 antigen was observed after immunization with the HIV-1 immunogen (p = 0.02). A strong association was demonstrated between the HIV-1 immunizing antigen, HZ321, and native p24 antigen responses (r = 0.80, p < 0.0001). Furthermore, a strong association in terms of proliferative responses was demonstrated between HZ321 virus (clade A) responses and BAL virus (clade B) (r = 0.95, p < 0.0001) and clade E virus antigen (r = 0.92, p < 0.0001). Proliferative responses to HIV antigens also correlated with baseline CD4 counts. Taken together, these results support the specificity of immune responses induced by REMUNE (HIV-1 immunogen). The development of cross-reactive immune responses between clades and to the more conserved epitopes of the virus have implications in the development of therapeutic and prophylactic HIV vaccines.

Early clinical markers and CD4 percentage in subjects with human immunodeficiency virus infection.

In a clinical trial involving asymptomatic, HIV-seropositive subjects treated with either the HIV-1 immunogen (an inactivated, gp120-depleted HIV-1 virus in incomplete Freund's adjuvant) or an adjuvant control, we examined the relationship between changes in the percentage of CD4 cells over time and early clinical markers of HIV disease progression. Subjects who had an early clinical event were more likely to have a greater decline in the percentage of CD4 cells than those subjects who did not have a clinical event (p = 0.054). The greatest decline in CD4 percentage occurred within 10 weeks prior to a clinical event (mean 11% decrease from baseline). Subjects from the quartile with the greatest decline in CD4 percentage had a fivefold greater risk of having a clinical event than subjects from the quartile with the second largest decline (p = 0.045). These results demonstrate a relationship between changes in the percentage of CD4 cells and early clinical events. Further validation of this association may be useful in clinical monitoring and in evaluating therapies to treat HIV infection.