Low prevalence of hepatitis C virus infection in Amerindians from Western Venezuela

Previous studies have not found hepatitis C virus (HCV) infection in Amerindians from Western Venezuela. A survey of 254 Bari and Yukpa natives aged 10-60 years (mean +/- SD age = 35 +/- 5.4 years) from four communities, two Bari and two Yukpa, in this area were studied to assess the prevalence of antibodies to HCV (anti-HCV) and HCV RNA among these indigenous populations. Serum samples were examined initially for anti-HCV by a four generation enzyme-linked immunosorbent assay (ELISA). Reactive samples were then tested using a third generation recombinant immunoblot assay (RIBA-3). Viral RNA was investigated in all immunoblot-reactive samples by a nested polymerase chain reaction (PCR) method. Six (2.3%) of 254 natives were positive by ELISA, one (2.2%) of these reactive samples were positive by RIBA, and four (1.5%) were indeterminate. Only two (0.8%) were positive by PCR, corresponding to 1 (2.1%) of 47 inhabitants of a Yukpa community and to 1 (2.2%) of 45 subjects of a Bari community. Iatrogenic is thought to play a role in acquisition of the infection. The findings indicate a HCV focus of low endemicity and are compatible with a low degree of exposures of the natives to the virus. Studies are necessary to assess the risk factors for infection in these Amerindians.     

Behavior of antithrombin III isoforms on immobilized heparins. Evidence that the isoforms bind to different numbers of low-affinity heparin sites

Antithrombin III exists in plasma as major and minor isoforms differing in affinity for heparin. The nature of the binding of each purified isoform to immobilized heparins was investigated. Unfractionated, mixed-affinity heparin bound each isoform with both high affinity and concentration-dependent low affinity. The isoforms were resolved when filtered through low-affinity heparin (heparin repeatedly passed over immobilized antithrombin III) columns. Following chemical modification of a specific tryptophan residue required for heparin binding, each isoform failed to bind to either low-affinity or mixed-affinity heparin-agarose, but elution of the modified higher-affinity isoform was retarded on both gels. Because the modified lower-affinity isoform eluted with the similarly sized bovine serum albumin in these experiments, the difference in isoform affinity for heparin appears to be the result of a unique, secondary heparin-binding site in the higher-affinity isoform that can bind a heparin site with low affinity for antithrombin III. This interpretation was supported by the chromatographic behavior of the isoforms on mixed-affinity agarose during reverse gradient elution. Two other populations of each of the tryptophan-modified isoforms were identified. Since these isoforms bound tightly to mixed-affinity heparin-agarose but eluted at lower salt concentrations than the corresponding unmodified isoforms, both isoforms may contain additional secondary sites that interact weakly with heparin. A general model of heparin-antithrombin III interaction is proposed in which a high-affinity heparin site initially interacts with a primary site on antithrombin III. The subsequent conformational change leads to a cooperative, entropy-driven association between secondary sites on the protein and low-affinity sites on heparin, stabilizing antithrombin III in its activated form.     

Separation of Chiral Molecules: A Way to Homochirality

A mechanism for separating chiral molecules is proposed. The separation of two enantiomers in an aquifer is considered. The molecules are dragged in the aquifer porous medium by a flow of water or of another liquid. The molecule velocity is u=v/R, where v is the liquid velocity, and R is the retardation factor. The aquifer consists of two one-dimensional layers disposed in series. The layers differ by the retardation factor or by the liquid velocity. The enantiomer velocity is a function of the enantiomer concentrations. This function is different in the two layers. For certain values of the model parameters and when the molecules entering the aquifer are enantiomerically enriched or when the medium is chiral, the concentration of one enantiomer increases around the interface between the two layers and in one layer, whereas the concentration of the other enantiomer decreases. Enantiomer synthesis and decay are not taken into account. The needed values of the parameters can be obtained when the enantiomers are moved by an alternating liquid flow, and the retardation factor oscillates in synchronism with the alternating liquid flow. The parameters of the model are then understood as quantities averaged over one oscillation period. The equations that give the values of the stationary concentrations of the enantiomers are found. The evolution of the enantiomer concentrations is determined by numerically solving a system of two nonlinear advection-dispersion equations. The proposed mechanism may have played a role in the emergence of biomolecular homochirality.     

Beaching of the tunicate<Emphasis Type="Italic"> Salpa thompsoni</Emphasis> at high levels of suspended particulate matter in the Southern Ocean

A mass death event of the pelagic tunicate Salpa thompsoni, which occurred in April 2002 in Potter Cove near the Argentinean/German Antarctic station Jubany (62°14′S–58°40′W), King George Island, South Shetland Islands, is described. Salps appeared on the beach 2 days after very strong (>80 km.h⁻¹) winds were registered, which accumulated particulate material in the inner part of the cove and probably also re-suspended bottom sediments. The sharp increase in particulate-matter concentrations in Potter Cove caused clogging of salp mucous filtering nets, and probably a combination of clogging, winds and tides caused dying salps to be washed out onto the beach.     

Adult mouse kidneys become permissive to acute polyomavirus infection and reactivate persistent infections in response to cellular damage and regeneration.

Kidneys of newborn (but not adult) mice are normally high permissive for polyomavirus (Py) infection and readily establish persistent infections. We have proposed that ongoing cellular differentiation, which occurs in newborn mice, may be necessary for a high level of in vivo Py replication (R. Rochford, J. P. Moreno, M. L. Peake, and L. P. Villarreal, J. Virol. 66:3287-3297, 1992). This cellular differentiation requirement may also be necessary for the reactivation of a persistent Py kidney infection and could provide an alternative to the accepted view that reactivation results from immunosuppression. To examine this proposal, the ability of adult BALB/c mouse kidneys to support primary acute Py infection or to reactivate previously established persistent Py infections after kidney-specific damage was investigated. Kidney damage was induced by both chemical (glycerol, cisplatin, or methotrexate) and mechanical (through renal artery clamping to produce unilateral renal ischemia) treatments. We also examined the effects of epidermal growth factor (EGF), which enhances the rate of kidney regeneration, on Py replication. Using histopathologic techniques, in situ hybridization for Py DNA, and immunofluorescence for Py VP1 production, we established that both chemical damage and damage through renal artery clamping of adult kidneys promoted high levels of primary Py replication in these normally nonpermissive cells. This damage also promoted the efficient reactivation of Py replication from persistently infected kidneys, in the absence of immunosuppression. EGF treatment significantly increased acute Py replication and also reactivation in damaged kidneys. These results support the view that ongoing cellular division and differentiation may be needed both for high levels of acute Py replication and for reactivation of persistent infections in vivo.     

Intratumoral spread and increased efficacy of a p53-VP22 fusion protein expressed by a recombinant adenovirus

In vitro experiments have demonstrated intercellular trafficking of the VP22 tegument protein of herpes simplex virus type 1 from infected cells to neighboring cells, which internalize VP22 and transport it to the nucleus. VP22 also can mediate intercellular transport of fusion proteins, providing a strategy for increasing the distribution of therapeutic proteins in gene therapy. Intercellular trafficking of the p53 tumor suppressor protein was demonstrated in vitro using a plasmid expressing full-length p53 fused in-frame to full-length VP22. The p53-VP22 chimeric protein induced apoptosis both in transfected tumor cells and in neighboring cells, resulting in a widespread cytotoxic effect. To evaluate the anti-tumor activity of p53-VP22 in vivo, we constructed recombinant adenoviruses expressing either wild-type p53 (FTCB) or a p53-VP22 fusion protein (FVCB) and compared their effects in p53-resistant tumor cells. In vitro, treatment of tumor cells with FVCB resulted in enhanced p53-specific apoptosis compared to treatment with equivalent doses of FTCB. However, in normal cells there was no difference in the dose-related cytotoxicity of FVCB compared to that of FTCB. In vivo, treatment of established tumors with FVCB was more effective than equivalent doses of FTCB. The dose-response curve to FVCB was flatter than that to FTCB; maximal antitumor responses could be achieved using FVCB at doses 1 log lower than those obtained with FTCB. Increased antitumor efficacy was correlated with increased distribution of p53 protein in FVCB-treated tumors. This study is the first demonstration that VP22 can enhance the in vivo distribution of therapeutic proteins and improve efficacy in gene therapy.     

Keeping close to the river,shore and surface: the first marine migration of brown trout (Salmo trutta) and Arctic charr (Salvelinus alpinus) post-smolts

Acoustic telemetry was utilized to track 49 brown trout (Salmo trutta) and 37 Arctic charr (Salvelinus alpinus) first-time migrants of wild origin [post-smolts; mean L_F (fork length): 169 and 172 mm] in a large fjord in northern Norway. The S. trutta were registered at sea for more than twice the time of the S. alpinus (medians of 54 and 22 days, respectively). Both species were mostly detected near river mouths (>80% of detections) and almost exclusively spent their time (>95%) within the interior 18 km of the fjord. They were surface oriented, with most detections at <1 m depth and S. trutta deeper on average (median mean depths of 0.7 and 0.5 m, respectively). This study concludes that post-smolts of both species stay closer to the surface and to river mouths than larger veteran migrants. This study emphasizes the importance of river mouths and upper water layers for the survival of both species during their first marine migration.