Wendell D. Winters

BCG immunotherapy of bladder cancer: Inhibition of tumor recurrence and associated immune responses

Fifty‐one patients with confirmed bladder cancer have enrolled in a prospective evaluation of BCG immunotherapy. Following resection of existing tumors, patients were stratified according to tumor grade and number of previous recurrences and randomly assigned to control or BCG treatment groups. Immunotherapy consisted of six weekly administrations of Pasteur strain BCG using 120 mg intravesically and 5 mg percutaneously. Immunotherapy side effects were minimal and no patient required postponement of BCG treatments. Eleven control (46%) compared with five (22%) BCG‐treated patients had tumor recurrence (P = 0.078, χ2). Prolongation of the disease‐free interval with BCG treatment was significant at the P = 0.016 level by Wilcoxon analysis. Four control and two BCG‐treated patients had multiple recurrences. Comparing total episodes of recurrence, nineteen of 79 (24%) control and eight of 85 (7%) BCG group cystoscopic examinations revealed tumor (P = 0.006, χ2). Immunologic correlates of response to immunotherapy were not statistically significant since only five BCG‐treated patients had tumor recurrence. However, four of these five patients evidenced impaired LIF response to PPD at the time of tumor recurrence, and impairment of skin test reactivity and BCG humoral antibody response were more commonly seen in this subgroup of patients.

In Vitro Exposure to Electromagnetic Fields: Changes in Tumour Cell Properties

Two human colon cancer cell lines, Colo 205 and Colo 320 DM, have been studied for their responses to 60 Hz-generated electromagnetic fields (EMF) using soft agar cloning and monoclonal antibody binding assays to assess exposure-induced changes. Cellular responses have been studied after 24 h continuous exposure of cells concurrently to four experimental conditions; i.e. no EMF (E-M-), magnetic field only (M+, 1.0 G rms), electric field only (E+, 300 mA/m2 rms), and combined electric plus magnetic fields at these intensities (E+M+). Under these conditions, both cell lines demonstrated significantly increased colony formation in soft agar and increased expression of tumor associated antigens after exposure to E+M+ and to M+ as compared to unexposed controls.

Antiviral Activity of Aloe Extracts against Cytomegalovirus

Substances extracted from the leaves of aloe plants have been reported to have antiviral effects against enveloped viruses in in vitro test systems. In the present studies, we have assessed the antiviral activity of partially purified extracts prepared from the gel portion of leaves of Aloe barbadensis Miller against human cytomegalovirus (CMV) by plaque inhibition tests (PIT), flow cytometry and morphometry assays. Six test extracts of gel filet portions of aloe leaves were prepared; i.e. R1 from immature leaves harvested in the early summer, S1 from mature leaves harvested in the autumn, F1 from S1 after freezing at −20°C, and R2, S2 and F2 which were ethanol treated extracts of R1, S1 and F1, respectively. When test aloe extracts were added at various concentrations during the course of infection with CMV, R1, S1 and F2 at concentrations of 10−1 inhibited plaque formation. The addition of S1 to the medium between 12 and 36 h after initiation of CMV infection, a time of high DNA synthesis, caused the most effective plaque inhibition. Flow cytometric and morphometric analyses revealed no significant differences in aloe extract treated, CMV infected cells compared with non‐aloe treated CMV infected control cells. The results suggest that a major mechanism of inhibition of CMV infection by aloe extracts is through interference with DNA synthesis.

Increased antibiotic resistance of E. coli exposed to static magnetic fields

The present study demonstrates that exposure of bacteria to medium strength static magnetic fields can significantly alter antibiotic sensitivity. Cultures of Escherichia coli were exposed to fields produced by permanent magnets. Samples of bacterial cultures continuously growing in the presence and in the absence of static magnetic fields were left untreated or were treated with an antibiotic and measured at 45 min intervals for cell growth and survival. It was found that exposure of E. coli to the static fields significantly increased antibiotic resistance. Bioelectromagnetics 22:129-137, 2001. Published 2001 Wiley-Liss, Inc.

Sensitivity of Human Oral Tumor Cells to Human Adenovirus

Human cells in early passage cultures of benign oral tumors, normal oral tissues, normal lung tissues and, especially, in long-term established oral carcinoma cultures were highly susceptible to infection by human adenovirus types 5, 21, and 31. In contrast, replication of each adenovirus type was markedly limited in inoculated cells of newly-established oral squamous cell carcinoma cultures.

A new perspective of microbial survival and dissemination in a prospectively contaminated air-fluidized bed model

A major concern of users of air-fluidized beds has been the possibility that such beds might be a source of microbial contamination. The purpose of this series of prospective, controlled experiments was to measure quantitatively the dissemination and survival of Bacillus subtilis, a species of bacterium that forms desiccation-resistant spores, as it was associated with circulating and clumped microbeads after challenge in an air-fluidized bed operating under decontamination conditions of heating at 48 degrees C and microbead agitation by an air flow of 100% at 110 cu ft/min. Microbead samples collected after B. subtilis challenge from predesignated depths and locations within the air-fluidized bed at 0.25, 1, 2, 4, 24, and 48 hours were assayed for colony-forming units (CFU) of challenge bacteria by end point dilution and streak-plate assays. Results of three experiments with an average challenge of 1110 CFU of B. subtilis per gram of microbeads indicated that no more than 0.46 CFU of B. subtilis per gram of circulating microbeads and 0.78 CFU per gram of clumped microbeads were detected after 24 to 48 hours at decontamination conditions. Few microbes were detected during three control (sterile water) challenges of the air-fluidized bed operating at 33 degrees C with 95% to 100% air flow. This study has demonstrated that an air-fluidized bed operating at the described decontamination conditions for 24 to 48 hours caused significant decreases, averaging a thousandfold per gram, in levels of microbead-associated challenge bacteria.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of environmental level magnetic field exposures on transcription of CMV immediate early promoter DNA in a cell-free in vitro transcription system

Effects of environmental levels of magnetic fields (MFs) on RNA synthesis have been investigated by using a cell-free system for in vitro transcription. Transcription reaction mixtures containing CMV immediate early promoter DNA plus HeLa cell nuclear extracts were exposed to each of three different MF field strengths, i.e., 10, 50, and 100 microT. Each MF exposed extract was paired with a simultaneous sham-exposure control. The present results show no significant differences in amounts of RNA synthesis in extracts of MF exposed compared with that in the sham controls. This finding is in contrast to results of prior studies of DNA synthesis in cell-free systems that showed MF exposure effects. The results of the present cell-free system studies suggest that the marked differences of MF exposure effects on DNA and on RNA synthesis direct attention to the complexity involved in confirming significant effects of exposures to environmental levels of MFs in biosystems in vivo and in vitro.

Polypeptides of the Three Major Medicinal Aloes

Major polypeptide species of proteins have been identified and analysed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS‐PAGE) in fresh extracts of the gel portion of mature leaves of Aloe saponaria Haw, Aloe barbadensis Miller, and Aloe arborescens Miller var. natalensis Berger. Extracts of gel from freshly cut Aloe leaves were prepared by dissection, tissue disruption, ethanol extraction, differential centrifugation, and gel filtration methods. Comparisons of SDS‐PAGE profiles of Aloe leaf gels showed that all three medicinal Aloes share five major common polypetides. Other results indicated that Aloe saponaria and Aloe barbadensis share the most major polypeptides, i.e. 11, and Aloe barbadensis and Aloe arborescens the least, i.e. 5, while Aloe arborescens had three individual polypeptides not found in the other two species. This is the first report to describe the nature and types of polypeptides detected in extracts of leaf gel portions of mature Aloe plants of these three medicinal species. Accordingly, this report may be of considerable value in helping to identify and characterize those Aloe substances of medicinal importance which are associated with each or all of these medicinal Aloes.

Environmental magnetic fields change complementary DNA synthesis in cell-free systems.

A number of studies have shown that exposures to environmental magnetic fields (MFs) increase cellular transcription and enhance DNA synthesis. However, little is known about the basic mechanisms underlying specific biological responses to MFs. We looked directly at the effect of MFs by using a cell-free rabbit globin cDNA synthesis system. cDNA synthesis reaction mixtures were placed in each of four exposure conditions and were tested simultaneously. Condition one was the control, with no exposure during either single-strand or double-strand synthesis (M-M-). The second condition was MF exposure only during single-strand synthesis (M+M-). The third condition was exposure only during double-strand synthesis (M-M+). The fourth condition was exposure during both single-strand and double-strand synthesis (M+M+). Results showed that cDNA synthesis was affected by 5-100 microT, 60 Hz MFs. Double-strand cDNA synthesis increased with MF exposure only during double-strand synthesis reactions (M-M+), and the greatest increase of double-strand cDNA synthesis was detected when MF exposure was at 10 microT, only during double-strand synthesis. Double-strand cDNA synthesis decreased when only single-strand synthesis reactions were exposed (M+M-; 100 microT). An increase of cDNA synthesis caused increased synthesis of rabbit globin cDNA and large-sized molecules. These results suggest that exposure to MF induced structural changes of synthesized cDNA, therefore altering the amount of cDNA. Our results show that environmental MFs can significantly alter cDNA synthesis in a cell-free system.

Concomitant transfer of BCG-CW and canine virus antibodies from dams to pups.

Abstract Levels of antibodies against infectious canine hepatitis virus (ICHV) and Bacillus Calmette-Guerin cell wall (BCG-CW) antigens in sera from BCG-CW inoculated dams, uninoculated dams, neonate pups in litters born to these dams and normal female dogs were measured by solid phase bead-type radioimmunoassay (SBRIA) tests. Significantly higher levels of antibodies reactive with BCG-CW, but not ICHV, antigens were found in the sera from two day old pups born to BCG-CW inoculated, ICHV-immune dams than in sera from two day old pups born to ICHV-immune dams not inoculated with BCG-CW. These results demonstrate that maternal antibodies against antigens of a non-viable immunotherapeutically active preparation (BCG-CW), and against antigens of ICHV, a canine virus routinely used in vaccines, can be passively transferred from dams to newborn pups.