Biomedical subjects
S Hopkins
Publications and source records attributed to S Hopkins.
Prolonged therapy with the fusion inhibitor T-20 in combination with oral antiretroviral agents in an HIV-infected individual.
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Bilin organization in cryptomonad biliproteins.
The bilin organization of three cryptomonad biliproteins (phycocyanins 612 and 645 and phycoerythrin 545) was examined in detail. Two others (phycocyanin 630 and phycoerythrin 566) were studied less extensively. Phycocyanin 645 and phycoerythrin 545 were suggested to have one bilin in each monomeric (alphabeta) unit of the dimer (alpha2beta2) isolated from the others, and the remaining six bilins may be in pairs. One pair was found across the monomer-monomer interface of the protein dimer, and two identical pairs were proposed to be within the monomer protein units. For phycocyanin 612, a major surprise was that a pair of bilins was apparently not found across the monomer-monomer interface, but the remaining bilins were distributed as in the other two cryptomonad proteins. The effect of temperature on the CD spectra of phycocyainin 612 demonstrated that two of the bands (one positive and one negative) behaved identically, which is required if they are coupled. The two lowest-energy CD bands of phycocyanin 612 originated from paired bilins, and the two higher-energy bands were from more isolated bilins. The paired bilins within the protein monomers contained the lowest-energy transition for these biliproteins. Using the bilins as naturally occurring reporter groups, phycocyanin 612 was shown to undergo a reversible change in tertiary structure at 40 degrees C. Protein monomers were shown to be functioning biliproteins. A hypothesis is that the coupled pair of bilins within the monomeric units offers important advantages for efficient energy migration, and other bilins transfer energy to this pair, extending the wavelength range or efficiency of light absorption.
Syringe exchange and risk of infection with hepatitis B and C viruses.
The authors utilized a cohort study among Seattle injection drug users (IDUs) to assess whether participation in a syringe exchange program was associated with incidence of hepatitis B virus (HBV) and hepatitis C virus (HCV) infection. Susceptible IDU subjects (187 seronegative for antibody to HCV, and 460 seronegative for core antibody to HBV) were identified in drug treatment, corrections, and social service agencies from June 1994 to January 1996, and followed for seroconversion one year later. The subjects included in the analysis were Seattle-King County (Washington State) area IDUs enrolled in a larger multipurpose cohort study, the Risk Activity Variables, Epidemiology, and Network Study (RAVEN Study). There were 39 HCV infections (20.9/100/year) and 46 HBV infections (10.0/100/year). There was no apparent protective effect of syringe exchange against HBV (former exchange users, relative risk (RR) = 0.68, 95% confidence interval (CI) 0.2-2.5; sporadic exchange users, RR = 2.4, 95% CI 0.9-6.5; regular users, RR = 1.81, 95% CI 0.7-4.8; vs. RR = 1.0 for nonusers of the exchange; adjusted for daily drug injection). Neither did the exchange protect against HCV infection (sporadic users, RR = 2.6, 95% CI 0.8-8.5; regular users, RR = 1.3, 95% CI 0.8-2.2; vs. RR = 1.0 for nonusers; adjusted for recent onset of injection and syringe sharing prior to enrollment). While it is possible that uncontrolled confounding or other bias obscured a true beneficial impact of exchange use, these data suggest that no such benefit occurred during the period of the study.
Strategies to increase enrollment in children's health insurance programs: a report of the New York Academy of Medicine.
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A discussion of the legal aspects of female genital mutilation.
The purpose of this paper is to examine the position of the nurse/midwife in the United Kingdom when involved with the care of a woman or female child who has suffered genital mutilation, which is an illegal practice in this country and most other areas of the world. The types of circumcision commonly practised are introduced, the prevalent reasons for the continuation of the practice among certain ethnic groups are presented, and the range of issues to be considered by the nurse is examined. These include international and national legal aspects which do not exist in isolation and are considered in context with cultural, medical, human rights and gender issues. Nursing legal issues include child protection, consent, advocacy and confidentiality, which invoke the Code Of Professional Conduct of the United Kingdom Central Council for Nursing Midwifery & Health Visiting. Midwives and nurses working in the field of gynaecology have raised questions regarding possible courses of action to take when presented with this issue. Increased knowledge can help to inform those decisions. Therefore, implications for future practice are addressed, together with recommendations to assist nurses with decision making when faced with this scenario in the future.
The nature of the attenuation of Salmonella typhimurium strains expressing human papillomavirus type 16 virus-like particles determines the systemic and mucosal antibody responses in nasally immunized mice.
We have recently shown by using a recombinant Salmonella typhimurium PhoPc strain in mice the feasibility of using a Salmonella-based vaccine to prevent infection by the genital human papillomavirus type 16 (HPV16). Here, we compare the HPV16-specific antibody responses elicited by nasal immunization with recombinant S. typhimurium strains harboring attenuations that, in contrast to PhoPc, are suitable for human use. For this purpose, chi4989 (Deltacya Deltacrp) and chi4990 [Deltacya Delta(crp-cdt)] were constructed in the ATCC 14028 genetic background, and comparison was made with the isogenic PhoPc and PhoP- strains. Although the levels of expression of HPV16 virus-like particle (VLP) were similar in all strains, only PhoPc HPV16 induced sustained specific antibody responses after nasal immunization, while all strains induced high antibody responses with a single nasal immunization when an unrelated viral hepatitis B core antigen was expressed. The level of the specific antibody responses induced did not correlate with the number of recombinant bacteria surviving in various organs 2 weeks after immunization. Our data suggest that the immunogenicity of attenuated Salmonella vaccine strains does not correlate with either the number of persisting bacteria after immunization or the levels of in vitro expression of the antigen carried. Rather, the PhoPc phenotype appears to provide the unique ability in Salmonella to induce immune responses against HPV16 VLPs.
Quantitation of a 36-amino-acid peptide inhibitor of HIV-1 membrane fusion in animal and human plasma using high-performance liquid chromatography and fluorescence detection.
Selective extraction of a 36-amino-acid peptide (DP-178, T20, pentafuside) from the protein matrices of animal and human plasma was achieved using acetonitrile containing 1% trifluoroacetic acid and 1% n-nonyl-beta-D-glucopyranoside. The peptide concentration of the extract was measured using reversed-phase high-performance liquid chromatography (RP-HPLC) and fluorescence detection. The eluent was excited at 280 nm and the intrinsic fluorescence signal was collected at 350 nm. Recovery of T20 from the plasma matrices was 75% (mouse), 60% (rat), 50% (cynomolgus monkey), and 55% (human) based on parallel-processed aqueous T20 standard solutions. The fluorescence peak area vs. concentration of T20 was linear in the range 4-160 ng/ml based on the final solute concentration in the HPLC vial, corresponding to original plasma concentrations of 100-4000 ng/ml. Experiments with truncated analogs of T20 demonstrate that this assay offers the advantage of detecting metabolites attributable to bio-transformation degradation processes differing by as little as one amino acid from the original peptide.
Phycoerythrin 545: monomers, energy migration, bilin topography, and monomer/dimer equilibrium.
Phycoerythrin 545 was isolated having an alpha2beta2 (dimer) protein structure at pH 6.0 and 2 g/L protein concentration with eight bilin chromophores. Monomers (alphabeta) were produced by lowering the protein concentration to 0.15 g/L and the pH to 4.5. Dimer dissociation was monitored by dynamic light scattering and gel-filtration column chromatography. Monomers were stable and had bilin optical spectra different from the alpha2beta2 dimers, although they have very similar protein secondary structures. The optical spectra of phycoerythrin 545 showed four types of behavior with temperature: 10-20 degrees C, dimers; 40-50 degrees C, dimers/monomers; 60 degrees C, nearly fully disordered; 70 degrees C, disordered alpha and beta polypeptides. At 40 degrees C, the protein dissociated partially to monomer, which could be totally reversed to dimers at 20-25 degrees C. The visible circular dichroism difference spectrum for the protein dimers minus monomers exhibited positive and negative bands--such spectra may indicate exciton splitting between closely-spaced bilins. Circular dichroism also revealed a spectrum suggesting exciton coupling for the second excited state of the bilins. Ultrafast fluorescence using a two-photon method showed the fastest time for protein dimers to be 2. 4 ps and monomers had a 39-ps lifetime. Phycocyanin 645 was found to have a 550-fs lifetime.
Safety of trovafloxacin in treatment of lower respiratory tract infections.
Safety and toleration of oral trovafloxacin has been assessed in Phase III trials in patients with acute lower respiratory tract infections. Patients were treated orally with either trovafloxacin 100 or 200 mg (n=881) or a comparator (500 or 1000 mg amoxicillin, or 625 mg amoxicillin/clavulanic acid; n = 593). Adverse events were recorded in 112 (12.7%) trovafloxacin- and 74 (12.5%) comparator-treated patients. Frequency of effects on the autonomic nervous, musculoskeletal, respiratory, special senses, urinary and reproductive systems was <1%. Photosensitivity reactions were not reported in trovafloxacin-treated patients. Central or peripheral nervous system adverse effects (headache and dizziness) were slightly more common in trovafloxacin-treated patients (4.4% vs 1.9%). Patients treated with comparators experienced gastrointestinal events more frequently (6.1% vs 8.3%). Comparable incidences of adverse events were reported in patients > or = 65-years-old. Most events were mild to moderate in severity. Treatment was discontinued because of an adverse event in 18 (2%) trovafloxacin- and four (0.7%) comparator-treated patients. Despite the high prevalence of risk factors, serious adverse events were rare and the mortality rate over the 35-day study period was low: trovafloxacin 0.8%, comparator agents 1.5%. Laboratory test abnormalities were recorded in less than 1% of patients in either treatment group.
Safety and tolerability of intravenous-to-oral treatment and single-dose intravenous or oral prophylaxis with trovafloxacin.
BACKGROUND: The new fourth-generation fluoroquinolone, trovafloxacin, is active in vitro against gram-positive and gram-negative organisms, atypical pathogens, and anaerobes, and has pharmacokinetics permitting once-daily intravenous or oral dosing. Safety/tolerability data from phase II/III clinical trials of sequential intravenous alatrofloxacin to oral trovafloxacin and single-dose intravenous or oral prophylaxis are summarized. METHODS: All trials were double-blind, randomized, and multicenter. In multidose trials with alatrofloxacin, 1,257 patients requiring initial intravenous therapy received once-daily alatrofloxacin (trovafloxacin prodrug, 200 or 300 mg/day) followed by oral 200 mg/day trovafloxacin. An additional 444 patients received a single alatrofloxacin or trovafloxacin dose prophylactically for surgical procedures. RESULTS: Therapy with alatrofloxacin was well tolerated. The most common treatment-related adverse events in studies of intravenous alatrofloxacin followed by oral trovafloxacin were nausea, headache, insertion site reaction, and dizziness. In single-dose intravenous or oral prophylaxis studies, insertion-site reaction, pruritus, and insertion-site pain were the most common treatment-related adverse events. No serious quinolone toxicity or drug interactions were reported. The incidence of serious treatment-related adverse events was < 1% in both the alatrofloxacin and comparator groups. In comparative trials, mortality due to all causes after trovafloxacin was similar to that after comparative agents; there was no mortality related to trovafloxacin administration. CONCLUSION: In this large patient sample, intravenous alatrofloxacin followed by oral trovafloxacin was safe and well tolerated. Serious adverse events, such as phototoxicity, cardiovascular toxicity, and hemolytic anemia associated with older fluoroquinolones, were not noted with trovafloxacin. No interactions of trovafloxacin with other drugs were reported. The safety and tolerability of trovafloxacin, along with in vitro activity against key pathogens and pharmacokinetics permitting once-daily administration, support its oral and intravenous use in patients with obstetric, gynecologic, and intra-abdominal infections as well as for prophylaxis of surgical infection.
Potent suppression of HIV-1 replication in humans by T-20, a peptide inhibitor of gp41-mediated virus entry.
T-20, a synthetic peptide corresponding to a region of the transmembrane subunit of the HIV-1 envelope protein, blocks cell fusion and viral entry at concentrations of less than 2 ng/ml in vitro. We administered intravenous T-20 (monotherapy) for 14 days to sixteen HIV-infected adults in four dose groups (3, 10, 30 and 100 mg twice daily). There were significant, dose-related declines in plasma HIV RNA in all subjects who received higher dose levels. All four subjects receiving 100 mg twice daily had a decline in plasma HIV RNA to less than 500 copies/ml, by bDNA assay. A sensitive RT-PCR assay (detection threshold 40 copies/ml) demonstrated that, although undetectable levels were not achieved in the 14-day dosing period, there was a 1.96 log10 median decline in plasma HIV RNA in these subjects. This study provides proof-of-concept that viral entry can be successfully blocked in vivo. Short-term administration of T-20 seems safe and provides potent inhibition of HIV replication comparable to anti-retroviral regimens approved at present.
Mice are protected from Helicobacter pylori infection by nasal immunization with attenuated Salmonella typhimurium phoPc expressing urease A and B subunits.
Live Salmonella typhimurium phoPc bacteria were tested as mucosal vaccine vectors to deliver Helicobacter pylori antigens. The genes encoding the A and B subunits of H. pylori urease were introduced into S. typhimurium phoPc and expressed under the control of a constitutive tac promoter (tac-ureAB) or a two-phase T7 expression system (cT7-ureAB). Both recombinant Salmonella strains expressed the two urease subunits in vitro and were used to nasally immunize BALB/c mice. The plasmid carrying cT7-ureAB was stably inherited by bacteria growing or persisting in the spleen, lungs, mesenteric or cervical lymph nodes, and Peyer's patches of immunized mice, while the plasmid carrying tac-ureAB was rapidly lost. Spleen and Peyer's patch CD4+ lymphocytes from mice immunized with S. typhimurium phopc cT7-ureAB proliferated in vitro in response to urease, whereas cells from mice given S. typhimurium phoPc alone did not. Splenic CD4+ cells from mice immunized with phoPc cT7-ureAB secreted gamma interferon and interleukin 10, while Peyer's patch CD4+ cells did not secrete either cytokine. Specific H. pylori anti-urease immunoglobulin G1 (IgG1) and IgG2A antibodies were detected following immunization, confirming that both Th1- and Th2-type immune responses were generated by the live vaccine. Sixty percent of the mice (9 of 15) immunized with S. typhimurium phoPc cT7-ureAB were found to be resistant to infection by H. pylori, while all mice immunized with phoPc tac-ureAB (15 of 15) or phoPc (15 of 15) were infected. Our data demonstrate that H. pylori urease delivered nasally by using a vaccine strain of S. typhimurium can trigger Th1- and Th2-type responses and induce protective immunity against Helicobacter infection.
Delivery systems and adjuvants for vaccination against HIV.
Epidemiologic studies have revealed that HIV-1 infections occur through contact with contaminated blood or during unprotected vaginal or anal intercourse. Hence, to protect against HIV infection, vaccines should induce both mucosal and systemic immune responses. We present a brief review of the different delivery systems and adjuvants which can be used to elicit mucosal immune responses. Oral or nasal administration of recombinant Salmonella vaccines can induce both mucosal and systemic immune responses against the carried antigen. The oral delivery of mucosal adjuvants (such as cholera toxin) in association with antigens has been shown to enhance mucosal and systemic immune responses against them. Recently developed vaccination strategies using naked DNA or recombinant adenovirus are also discussed.
Lack of an epidemiologic link between Kaposi's sarcoma and CNS lymphoma in patients with AIDS.
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Perioperative vasopressin secretion treated by demeclocycline.
The purpose of this study was to evaluate the perioperative effects of demeclocycline on vasopressin (VP) in patients undergoing surgery, specifically coronary artery bypass grafting (CABG). This was a prospective, double-blind placebo-controlled clinical study using human subjects in a 575-bed tertiary care teaching community hospital. Thirty patients (20 males and 10 females) undergoing elective CABG over a 6-month period were randomized preoperatively to receive either demeclocycline or a placebo. Each patient received either a total of 1200 mg daily of demeclocycline or a placebo beginning 5 days preoperatively and continuing through postoperative day 2. Urine and serum osmolality, electrolytes, and VP levels were measured daily. Perioperative VP levels were significantly higher (P = 0.05) in the demeclocycline group despite decreased VP activity. The postoperative serum sodium and osmolality remained normal in the demeclocycline group and significantly decreased in the placebo group (P < 0.01). The urine osmolality increased significantly in the placebo group (P = 0.04) on postoperative day 1. We conclude that perioperative administration of demeclocycline reliably inhibits the effects of increased VP secretion commonly seen in patients undergoing CABG procedures. Applying these findings to surgical patients who are at increased risk of complicated fluid and electrolyte problems requires further study.
Biological activity and brain actions of recombinant rat interleukin-1alpha and interleukin-1beta.
IL-1alpha and IL-1beta have potent effects on the central nervous system resulting in fever, activation of the hypothalamic-pituitary-adrenal axis and behavioural depression. These effects have mainly been studied in rats, using recombinant human and mouse IL-1. Because IL-1alpha and IL-1beta show some species specificity in the potency of their biological activities, the objective of the present work was to directly compare the effects of recombinant rat IL-1alpha and IL-1beta in the rat system as a first step to dissect out the mechanisms that are involved in these effects. In vitro, recombinant rat IL-1alpha and IL-1beta bound with the same affinity as human IL-1 to the rat insulinoma Rin m5F cell line that mainly expresses type I IL-1 receptors. This binding activated IL-1 receptors, as shown by induction of the synthesis of TNF-alpha mRNA. In vivo, recombinant rat IL-1alpha and IL-1beta enhanced body temperature, increased plasma levels of corticosterone and ACTH, and depressed social behaviour. All these effects were obtained at doses 100-1,000 fold lower when IL-1 was injected centrally than when it was administered peripherally, indicating that they are centrally mediated. The relative potencies of recombinant rat IL-1alpha and IL-1beta were not the same depending on the endpoint and the route of injection, indicating that different mechanisms are likely to be involved in the various effects of IL-1 on the brain.
Hybrid hepatitis B virus core antigen as a vaccine carrier moiety. II. Expression in avirulent Salmonella spp. for mucosal immunization.
Hepatitis B virus (HBV) core antigen (HBcAg) is a highly immunogenic subviral particle. We and others have defined insertion sites for heterologous epitopes and successfully used hybrid particles to generate B and T cell immunity (reviewed in: Schödel et al. 1994a, 1995). Here we shall review recent progress in constructing avirulent Salmonella spp. expressing hybrid HBcAg particles carrying different epitopes. Hybrid HBcAg particles carrying virus neutralizing epitopes of the hepatitis B virus pre-S region or repeat epitopes of plasmodial circumsporozoite antigens were previously described (Schödel et al. 1992, 1994b). Salmonella spp. can be attenuated by defined genetic means so that they become avirulent, yet preserve invasiveness after oral uptake. Hybrid HBcAg-pre-S particles were expressed in Salmonella typhimurium and S. typhi vaccine strains. A single oral immunization of mice with such live recombinant S. typhimurium strains elicited a high titered serum anti-pre-S1 IgG response. Similarly, circumsporozoite repeat epitopes of three different malaria parasites were expressed as HBcAg-CS hybrids in recombinant S. spp. and were found to be highly immunogenic after oral immunization. To analyze mucosal immune responses, BALB/c mice were immunized with recombinant phoPc S. typhimurium expressing HBcAg by various mucosal routes (Hopkins et al., 1995). All routes of immunization resulted in high titered serum and local antibodies against HBcAg and S. typhimurium LPS. However, nasal immunization was most efficient in generating pulmonary IgA and rectal immunization in eliciting rectal IgA, suggesting some compartmentalization of the mucosal immune response.