J P Hansen

Clinical correlation of variations in the internal transcribed spacer regions of rRNA genes in Pneumocystis carinii f.sp. hominis.

ObjectivesTo analyse the importance of sequence variations in the internal transcribed spacer (ITS) regions 1 and 2 of the nuclear rRNA operon in AIDS patients with Pneumocystis carinii pneumonia (PCP). Design and methodsITS 1 and 2 genotypes were determined in 162 bronchoalveolar lavage samples from 130 patients participating in a prospective cohort study of PCP. ResultsA total of 49 different ITS genotypes were detected. ITS genotype was not associated with the clinical severity or outcome of PCP. In 37 of 162 (23%) samples infection with two or more genotypes was observed. A genotype switch was detected in six of 10 patients (60%) with recurrent episodes of PCP. However, genotype changes were also seen in 10 of 19 patients (53%) who had repeated bronchoscopies within the same episode of PCP. The same ITS type was observed twice in 13 (46%) of the 28 patients with repeat bronchoscopies during single or recurrent episodes of pneumonia, but in only 14 of 81 (17%) randomly selected pairs (P < 0.01). ConclusionAlthough the detection of ITS genotypes is not a random event, changes in genotype can be detected in a single episode of disease, with 23% of PCP patients being infected with more than one P. carinii genotype, thus complicating the use of this locus as a genetic marker to separate new infection from the reactivation of latent infection. ITS genotypes are not associated with the clinical severity of PCP.

Improved Humoral and Cellular Immune Responses Against the gp120 V3 Loop of HIV-1 Following Genetic Immunization with a Chimeric DNA Vaccine Encoding the V3 Inserted into the Hepatitis B Surface Antigen

The gp120‐derived V3 loop of HIV‐1 is involved in co‐receptor interaction, it guides cell tropism, and contains an epitope for antibody neutralization. Thus, HIV‐1 V3 is an attractive vaccine candidate. The V3 of the MN strain (MN V3) contains both B‐ and T‐cell epitopes, including a known mouse H‐2d‐restricted cytotoxic T lymphocyte (CTL) epitope. In an attempt to improve the immunogenicity of V3 in DNA vaccines, a plasmid expressing MN V3 as a fusion protein with the highly immunogenic middle (pre‐S2 + S) surface antigen of hepatitis B virus (HBsAg) was constructed. Epidermal inoculation by gene gun was used for genetic immunization in a mouse model. Antibody and CTL responses to MN V3 and HBsAg were measured and compared with the immune responses obtained after vaccination with plasmids encoding the complete HIV‐1 MN gp160 and HBsAg (pre‐S2 + S), respectively. DNA vaccination with the HIV MN gp160 envelope plasmid induced a slow and low titred anti‐MN V3 antibody response at 12 weeks post‐inoculation (p.i.) and a late appearing (7 weeks), weak and variable CTL response. In contrast, DNA vaccination with the HBsAg‐encoding plasmid induced a rapid and high titred anti‐HBsAg antibody response and a uniform strong anti‐HBs CTL response already 1 week p.i. in all mice. DNA vaccination with the chimeric MN V3/HBsAg plasmid elicited humoral responses against both viruses within 3–6 weeks which peaked at 6–12 weeks and remained stable for at least 25 weeks. In addition, specific CTL responses were induced in all mice against both MN V3 and HBsAg already within the first 3 weeks, lasting at least 11 weeks. Thus, HBsAg acts as a ‘genetic vaccine adjuvant’ augmenting and accelerating the cellular and humoral immune response against the inserted MN V3 loop. Such chimeric HIV–HBsAg plasmid constructs may be useful in DNA immunizations as a ‘carrier’ of protein regions or minimal epitopes which are less exposed or poorly immunogenic.

Exploring the WEP with a pulsed cold beam of antihydrogen

The AEGIS experiment, currently being set up at the Antiproton Decelerator at CERN, has the objective of studying the free fall of antimatter in the Earth?s gravitational field by means of a pulsed cold atomic beam of antihydrogen atoms. Both duration of free fall and vertical displacement of the horizontally emitted atoms will be measured, allowing a first test of the WEP with antimatter.

Nosocomial Hiv-transmission in an outpatient clinic detected by epidemiological and phylogenetic analyses

OBJECTIVE To determine if a case of HIV-infection in a patient (GP) with common variable immunodeficiency, and with no known risk factors for HIV-infection, could be due to horizontal nosocomial transmission. METHODS For determination of time of transmission stored serum-samples from GP were analysed for HIV RNA content. Patient records were used to identify patients, who had received intravenous therapy on the same day as GP. Samples from GP and these possible source patients were identified and phylogenetic analyses of the env, gag and RT-encoding region of pol were performed. Furthermore, routines in conjunction with intravenous therapy were examined. RESULTS We identified a patient (FDL) harbouring virus almost indistinguishable from the virus isolated from GP. The pairwise nucleotide distance between the C2-V3-C3 region of the env and gag sequences from the two patients were 1.9 and 0.9% respectively. In addition, GP harboured HIV RNA with a foscarnet resistance mutation further lending support to virus from the foscarnet-treated FDL being the source of the infection. Interestingly, GP experienced increases in immunoglobulin production after contracting the HIV-infection, and decreases after antiretroviral-induced viral suppression. A clinical procedure which, under stressful conditions, could lead to breaches in infection control measures was identified. The source of the infection was most likely a contaminated multidose vial. CONCLUSION Through epidemiological and phylogenetic analyses a case of horizontal nosocomial HIV-transmission was disclosed. Identification of multidose vials as possible vehicles for horizontal nosocomial transmission recently led to the recommendation of restriction of the use of multidose vials, a recommendation supported by the present study. The study underlies the importance of a constant survey of infection control precautions.

Pelagic food web processes in an oligotrophic lake

Major pelagic carbon pathways, including primary production, release of extracellular products (EOC), bacterial production and zooplankton grazing were measured in oligotrophic Lake Almind (Denmark) and in enclosures (7 m3) subjected to artificial eutrophication. Simultaneous measurements at three days interval of carbon exchange rates and pools allowed the construction of carbon flow scenarios over a nineteen day experimental period.The flow of organic carbon was dominated by phytoplankton EOC release, which amounted from 44 to 58% of the net fixation of inorganic carbon. Gross bacterial production accounted for 33 to 75% of the primary production. The lower values of EOC release (44%) and bacterial production (33%) were found in the enclosures with added nutrients. The release of recently fixed photosynthetic products was the most important source of organic carbon to the bacterioplankton. Uptake of dissolved free amino acids was responsible for 52 to 62% of the gross bacterial production. Thus, amino acids constituted a significant proportion of the EOC. Zooplankton (< 50 µm) grazing on algae and bacteria accounted only for a minor proportion of the particulate production in May. Circumstantial evidence is presented that suggests the chrysophycean alga Dinobryon was the most important bacterial remover.The results clearly demonstrated EOC release and bacterial metabolism to be key processes in pelagic carbon cycling in this oligotrophic lake.

Monte-Carlo simulations of proton aurora

Abstract The spreading of a proton beam in the upper atmosphere is calculated based onMonte-Carlo simulations. The transport of the atoms is modelled in a magnetic field with dipolestrength. Neuralisation, ionisation and excitation mechanisms of the incoming particles areincluded from collision cross-sections of protons and hydrogen with an effective N 2 atmosphere. Assuming an isotropic pitch angle distribution for the incoming protons, theirspreading in the upper atmosphere and the return flux of the charged and neutral component ofthe hydrogen beam has been calculated. Depending on energy and the tilt angle of the magneticfield about 10% of the incoming particles return from the atmosphere as ENA (Energetic NeutralAtoms). The ENA returning from the atmosphere show a source region below 500 km for theincoming high energy protons. For low energy protons, the ENA originate mainly from twodifferent regions, one around 700 km and the other at 400 km altitude, reflecting their sensitivityto several charge exchange processes.

The N-dimensional coulomb problem : Stark effect in hyperparabolic and hyperspherical coordinates

We calculate the first-order energy shifts for the N-dimensional hydrogen atom exposed to a static electric field. The results are compared with numerical diagonalization of the Hamiltonian in a finite basis. Using simple scaling relations, we show how corrections to arbitrarily high order may be obtained from known results for the three-dimensional Coulomb problem.

Exact nondipole Kramers-Henneberger form of the light-atom hamiltonian: an application to atomic stabilization and photoelectron energy spectra.

The exact nondipole minimal-coupling Hamiltonian for an atom interacting with an explicitly time- and space-dependent laser field is transformed into the rest frame of a classical free electron in the laser field, i.e., into the Kramers-Henneberger frame. The new form of the Hamiltonian is used to study nondipole effects in the high-intensity, high-frequency regime. Fully three-dimensional nondipole ab initio wave packet calculations show that the ionization probability may decrease for increasing field strength. We identify a unique signature for the onset of this dynamical stabilization effect in the photoelectron spectrum.

Interference effects in the ionization of H2 by fast charged projectiles

A theoretical investigation of the experimentally observed (Stolterfoht N et al 2001 Phys. Rev. Lett. 87 023201) interference effects in the double differential cross sections for ionization of the hydrogen molecule by fast ion impact is reported. The H2/2H cross section ratios as a function of the ejected electron velocity show an oscillating pattern, for which Stolterfoht et al propose a formula C + G sin(k D)/(k D), where k is the electron momentum and D the internuclear separation in H2. Our analysis in its simplest form leads instead to a formula C + G sin(k|| D)/(k|| D) where k|| is the component of k parallel to the projectile velocity. The presented theoretical model thus explains why at 90° the interference pattern will be strongly suppressed. In addition to the simplified analysis a numerical evaluation of a more accurate model is presented, confirming the latter qualitative prediction.

Mutations in CCR5-coding sequences are not associated with SIV carrier status in African nonhuman primates.

African monkeys can be naturally infected with SIV but do not progress to AIDS. Since mutations in the human CCR5 gene have been shown to influence susceptibility to HIV infection and disease progression, we have now investigated whether mutations in CCR5-coding sequences in African nonhuman primates can explain species-specific differences in susceptibility to lentiviral infection. The animals studied comprise chronically infected monkeys corresponding to four natural hosts of SIV (Cercopithecus aethiops, Cercopithecus pygerythrus, Cercopithecus sabaeus, and Cercopithecus tantalus), noninfected animals from three species that are known to be susceptible to SIV infection (Cercopithecus patas, Cercopithecus Ihoesti, and Pan troglodytes), and monkeys of six species that do not carry SIV in the wild (Cercocebus galeritus, Cercocebus aterrimus, Cercopithecus ascanius, Cercopithecus nictitans, Cercopithecus neglectus, and Cercopithecus cephus). We observed a high degree of genetic divergence among the species. The rate of accumulation of amino acid mutations was, however, not higher in SIV carriers than in other nonhuman primates. No homozygous premature stop codons, deletions, or frameshift mutations were detected. In at least two animals, one infected AGM (Cercopithecus tantalus) and one noninfected monkey (Cercocebus aterrimus), the CCR5 alleles identified encode functional proteins, as they were identical in terms of amino acid sequence to that of functional CCR5 reported in the literature. We found no other consistent differences in the genetic variability of CCR5-coding sequences between the nonhuman primates that are carriers of SIV and those that are not.