The need for a transparent, ethical, and successful relationship between academic scientists and the pharmaceutical industry: a view of the Group for the Respect of Ethics and Excellence in Science (GREES).

UNLABELLED: This paper provides recommendations for fair and unbiased relationship between academic scientists and the pharmaceutical industry. INTRODUCTION: Real or perceived problems in the relationship between academics and the industry have been the subject of much recent debate. It has been suggested that academic clinicians should sever all links with the industry-a view that is rarely challenged. METHODS: Academic experts and members of the pharmaceutical industry were invited to an expert consensus meeting to debate this topic. This meeting was organized by the Group for the Respect of Ethics and Excellence in Science. Conflict of interest, competing interest, right and duties of academic scientist, authorship, and staff and student education were discussed. RESULTS: Guidelines for a transparent, ethical, strong, and successful partnership between the academic scientist and the pharmaceutical industry have been provided. CONCLUSIONS: The Group support interactions between the industry and clinicians provided that it is transparent and ethical.

Conditions for production, and some characteristics, of mycobacterial growth inhibitory factor produced by spleen cells from mice immunized with viable cells of the attenuated H37Ra strain of Mycobacterium tuberculosis.

Mycobacterial growth inhibitory factor (MycoIF), found in supernatant fluids of mouse spleen cell cultures that have been stimulated in vitro with homologous antigen, inhibited the intracellular multiplication of virulent tubercle bacilli within normal mouse peritoneal macrophages in vitro. Antigenically stimulated H37Ra-immunized mouse spleen cells required 72 h of incubation to produce supernatant fluids that would cause intracellular inhibition. Supernatant fluids from 48-h mouse spleen cell cultures were not able to produce intracellular inhibition. Investigation of the culture conditions showed that at lease 1.0% human serum was required in the tissue culture medium for the production of MycoIF by spleen cells from immunized mice. MycoIF activity was noted only in supernatant fluids from spleen cell cultures incubated with antigen for 72 h. MycoIF was nondialyzable and unaffected by freezing, lyophilization, or incubation at 60 C for 30 min. However, MycoIF was inactivated after incubation at 80 C for 30 min. MycoIF was unaffected by low hydrogen ion concentrations (pH 7 to 12), but exposure to higher hydrogen ion concentrations (pH 6, pH 5) significantly decrease MycoIF activity, and exposure to pH 4 to 2 abolished all activity. Supernatant fluids diluted 1:32 were still able to produce significant intracellular inhibition of growth of virulent tubercle bacilli.

Molecular weight and other characteristics of mycobacterial growth inhibitory factor produced by spleen cells obtained from mice immunized with viable attenuated mycobacterial cells.

Exposure of mycobacterial growth inhibitory factor (MycoIF) to trypsin, chymotrypsin, or neuraminidase decrease its ability to produce intracellular inhibition of mycobacterial growth within macrophages, suggesting that MycoIF was a glycoprotein. MycoIF was unaffected by deoxyribonuclease or ribonuclease. Supernatant fluids from antigenically stimulated H37Ra-immunized mouse spleen cells exposed to puromycin were unable to produce significant intracellular inhibition. This indicated that the presence of MycoIF activity in supernatant fluids required protein synthesis. The filtration of MycoIF-containing supernatant fluids on Sephadex G-150 demonstrated that significant MycoIF activity appeared only in those fractions which eluted on the downward side of the serum albumin peak. Based on protein standards filtered through the Sephadex gel, the molecular weight of MycoIF was calculated to be between 20,000 and 35,000. These calculations assumed that MycoIF is a globular protein. Attempts to purify MycoIF by anion exchange chromatography (diethylaminoethylcellulose) was not successful.

Macrophage migration inhibitory activity of mycobacterial growth inhibitory factor and the effect of a number of factors on mycobacterial growth inhibitory factor activity.

Mycobacterial growth inhibitory factor (MycoIF), which inhibits the intracellular multiplication of virulent tubercle bacilli within normal peritoneal macrophages in vitro, was tested for its ability to inhibit the migration of normal peritoneal exudate cells. The migration of peritoneal exudate cells was not inhibited by MycoIF. It was also shown that normal peritoneal macrophages infected with virulent Mycobacterium tuberculosis, strain H37Rv, required 72 h of incubation with spleen cell culture supernatant fluids containing MycoIF in order to inhibit intracellular bacillary multiplication. Treatment of infected macrophages with trypsin before their exposure to MycoIF abolished the ability of MycoIF to inhibit intracellular mutiplication of tubercle bacilli. Incubation of infected macrophages with goat anti-mouse globulin before their exposure to MycoIF also blocked the action of MycoIF.

Influence of the interdigestive myoelectric complex on enteric flora in the rat.

This study was designed to define the role of the interdigestive myoelectric complex in small intestinal bacteriostasis. In rats, six monopolar electrodes were surgically sewn to the small intestine at equal intervals. One week later myoelectric activity was recorded. Under different experimental conditions, segments of duodenum and ileum were cultured quantitatively, both aerobically and anaerobically. Five groups of 6 electrode-equipped animals each were studied after an overnight fast: rats in which (a) the interdigestive myoelectric complex was present, (b) the interdigestive myoelectric complex was disrupted for 6 h using morphine sulfate, (c) the interdigestive myoelectric complex was disrupted for 15 h using morphine sulfate, (d) the interdigestive myoelectric complex was disrupted for 15 h using phenylephrine, and (e) the interdigestive myoelectric complex returned after 15 h of morphine sulfate effect. In control rats and during baseline records before drug administration in the other four groups, the interdigestive myoelectric complex was present. Activity fronts cycled at regular intervals in the proximal small intestine and moved aborally. Activity fronts disappeared following both morphine and phenylephrine, with varying degrees of inhibition of spike activity. Titers of microorganisms increased after 6 h, becoming statistically significant at 15 h; this effect was seen with both drugs. However, titers were similar to controls in groups 5. These results show that the interdigestive myoelectric complex is an important regulator at bacterial growth in the small intestine.

Methicillin-resistant Staphylococcus aureus susceptibility testing with Abbott MS-2 system.

The antimicrobial susceptibilities of 100 methicillin=resistant Staphylococcus aureus strains were concurrently determined by the Abbott MS-2 System and by the standard disk diffusion method. Agreement between the two methods was 94% or greater for all of the antibiotics tested except for methicillin and gentamicin. This study indicates that the Abbott MS-2 cannot be relied upon for detection of methicillin resistance in clinical S. aureus isolates.

Employment of tuberculostasis in serum-agar medium for the study of production and activity of Mycobactin.

Mycobactin (M), an iron-chelating product of tubercle bacilli, neutralized serum tuberculostasis by removing growth-essential iron from transferrin (Tr) and supplying the metal to the bacteria. The competition for iron between Tr and M has been demonstrated by the agar-plate diffusion test. This test is suitable not only for the study of Tr-iron-M interplay but also for the evaluation of serum tuberculostasis. Extremely poor solubility of M in water and consequently its association with lipoidal cell wall of tubercle bacillus was overcome by the use of water-dispersible and surface-active Tween 80. The addition of Tween 80 to culture media insured the presence of M in spent media; otherwise M was extracted from bacillary cells with a solution of Tween 80 or a mixture of ethanol and Tween 80. Although M was produced irrespective of the amount of iron present in culture medium, its production in iron-poor medium was more prolific than in iron-rich medium. M extracted from BCG or H(37)Rv cells neutralized serum tuberculostasis as effectively for the homologous as for heterologous strains. However, the extract of virulent bacilli was much more active in the neutralization than similar extract prepared from attenuated cells; whether this difference is of quantitative or qualitative nature remains to be determined.

Murine antibody-dependent cellular cytotoxicity to herpes simplex virus-infected target cells.

Freshly collected peritoneal cells (PC) and cultured spleen cells (SC) (but not fresh SC) from nonimmune mice could mediate antibody-dependent cellular cytotoxicity (ADCC) against herpes simplex virus (HSV)-infected cells in the presence of mouse or human sera containing antibody to HSV. PC also demonstrated variable natural killer cell cytotoxicity to infected cells. Both PC and cultured SC required high concentrations of antibody and high effector to target cell ratios for optimal ADCC. The time kinetics of the reaction appeared to depend on the state of activation of the effector cells. In both PC and SC populations, ADCC activity was limited to adherent cells, and was profoundly inhibited by particulate latex or silica. The murine effector cell found in PC and SC able to mediate ADCC to HSV-infected cells appears to be a macrophage.

Effect of herpes simplex virus infection on murine antibody-dependent cellular cytotoxicity and natural killer cytotoxicity.

Mice intraperitoneally inoculated with a sublethal dose of herpes simplex virus (HSV) produced immunoglobulin G antibody-dependent cellular cytotoxicity (ADCC) and radioimmunoassay (RIA) antibody as early as 3 days after infection. There was a rise in natural killer cytotoxicity (NKC) to infected and uninfected target cells 1 to 3 days postinfection mediated by nonadherent peritoneal cells (PC) in mice inoculated with HSV, but also with other substances commonly used in tissue culture media. HSV caused the highest and most consistent increase in NKC. PC-NKC, as ADCC, was inhibited by latex and silica, both macrophage inhibitors. PC-ADCC markedly declined 3 to 8 days after HSV inoculation. This was not due to a soluble or cellular suppressor factor, was not reversed by incubation or trypsin treatment of PC, was not associated with a change in PC Fc receptors, adherence, or acridine orange staining characteristics, and could not be induced by inactivated HSV. In vitro inoculation of PC with HSV similarly caused a reduction in the ability of PC to mediate ADCC to HSV-infected target cells. These data demonstrate the complex stimulatory and inhibitory interactions between virus and host defense mechanisms.

A rabbit model of intracerebral hematoma.

The epiphenomena that seem to cause deterioration and death after spontaneous interacerebral hematoma (SICH) might best be studied in an animal model. Therefore, the principles for developing such a model and techniques to study these phenomena were evaluated. Animals will tolerate injection of 3%-5% of their brain volume with a high proportion of clots. Fluorescein can be used to study the blood-brain barrier, and gravimetry to study edema. Others have found that injection of a paraffin/oil mixture can be employed for a control model. Refinement of the fluorescein technique, development of a primate model, and directions for future research are suggested.