The contributions of Dr. Roswell Park to epilepsy and spinal surgery.

ROSWELL PARK, M.D., (1852-1914) is remembered for founding the world's first cancer institute that now bears his name a century ago, The Roswell Park Cancer Institute, and for an unfortunate association with the mortal wounding of President William McKinley in Buffalo, NY, in 1901. Park's accomplishments as a pioneer American neurosurgeon have been overlooked. After Park was appointed as Chair of Surgery at the University of Buffalo in 1884, he became the first American surgeon to precisely localize and remove a posttraumatic epileptic focus in the absence of external scars in 1886. Park introduced American physicians and surgeons to David Ferrier's research on localization of cerebral cortical function and Victor Horsley's techniques for extirpating epileptic foci. In 1895, Park became the first American surgeon to successfully treat spina bifida. In the same year, he wrote the first American monograph on surgery of the head. Park's case reports of successful operations on patients deemed almost incurable reveal boldness and ingenuity. Park's untimely death truncated a promising career.

Contributions of the founders of Craig Colony to epileptology and public care of epileptics: 1890-1915.

Craig Colony in Sonyea, New York, was America's first comprehensive public epilepsy center. The background to its establishment (the first patients were admitted in 1896) and its role as a model for other institutions is described. The history of the first 25 years of the Colony is recounted and the contributions to epileptology, and the legacy to health care, of the founders--William Pryor Letchworth, Frederick Peterson, Roswell Park, William P. Spratling and Frederick Munson--are assessed.

The importance of Spratling.

William P. Spratling made important contributions to American epileptology at the beginning of this century. He was the first medical superintendent of Craig Colony for Epileptics from 1893 to 1908, cofounder and president of the National Association for the Study of Epilepsy, and first editor of its scholarly journal, Transactions. During his tenure at Craig Colony, Spratling established standards for safe and humane public care of epileptics. He started the first American residency training program emphasizing epileptology. Spratling conducted the first American multicenter research on the causes of death in epilepsy. The dosage of bromide therapy, which he empirically determined, remains correct. In his book Epilepsy and Its Treatment, Spratling substantiated the cortical origin theory of epilepsy developed by Jackson and Gowers. He was the first American to postulate and investigate a biochemical etiology of generalized seizures in the absence of anatomic lesions. Despite signal accomplishments, his untimely, tragic death may explain why he remains obscure.

William P. Letchworth: philanthropist and pioneer epileptologist.

Although neither a physician nor a scientist, William Pryor Letchworth significantly improved the care and treatment of epileptics at the beginning of this century. As commissioner of the New York State Board of Charities and later president, he established Craig Colony, America's first comprehensive epilepsy facility. In Care and Treatment of Epileptics, he summarized contemporary medical and social knowledge of epilepsy. As cofounder and president of the National Association for the Study of Epilepsy, he introduced from Europe his improvements of the colony plan of construction and financed Transactions, the society's scholarly journal. He combined a sensitivity to the needs of the unfortunate with the resolve of a successful businessman. Although William Pryor Letchworth is remembered for his philanthropy and the park in western New York that bears his name, his signal contributions to modern concepts of epilepsy are unknown to most physicians. This article will acquaint readers with the life and accomplishments of this philanthropist and pioneer epileptologist.

New soluble-formazan assay for HIV-1 cytopathic effects: application to high-flux screening of synthetic and natural products for AIDS-antiviral activity.

We have developed an effective and optimally safe microculture method for rapid and convenient assay of the in vitro cytopathic effects of human immunodeficiency virus (HIV-1) on human lymphoblastoid or other suitable host cells. The assay procedure is applicable to the evaluation of drug effects on in vitro infections induced directly in cultured host cells by cell-free HIV-1 or by coculture with H9 cells chronically infected with HIV-1. The assay uses a newly developed tetrazolium reagent that is metabolically reduced by viable cells to yield a soluble, colored formazan product measurable by conventional colorimetric techniques. This simple microassay minimizes the number of plate manipulations typically required with other assay methods and, coupled with computerized data collection and analysis, facilitates large-scale screening of agents for potential antiviral activity. To support and enhance the discovery of new anti-HIV-1 agents, the National Cancer Institute is offering investigators worldwide the opportunity to submit new candidate agents for anti-HIV-1 screening with this method.

Comparison of intrapulmonary, percutaneous intrathoracic, and subcutaneous models for the propagation of human pulmonary and nonpulmonary cancer cell lines in athymic nude mice.

The propagation efficiencies, growth patterns, histological appearances, and roentgenographic demonstration of tumors derived from six continuous human pulmonary tumor cell lines implanted intrathoracically (i.t.) and intrabronchially (i.b.) were compared with the conventional s.c. implantation method at three different tumor cell inocula (N = 184, i.b.; N = 185, i.t.; N = 180, s.c.). A tumor-related mortality of 100% was noted when the six different human lung tumor cell lines, including A549 adenocarcinoma, NCI-H125 adenosquamous carcinoma, NCI-H460 large cell undifferentiated carcinoma, NCI-H69 small cell carcinoma, and NCI-H358 and NCI-H322 bronchioloalveolar cell carcinomas, were implanted i.b. at a 1.0 x 10(6) tumor cell inoculum. A similar (92%) tumor-related mortality was observed when these same lung tumor cell lines were implanted i.t. at a 1.0 x 10(6) tumor cell inoculum (P greater than 0.10), whereas minimal (5%) tumor-related mortality was noted when cells from the six different cell lines were implanted s.c. (P less than 0.001). In addition, a dose-dependent, tumor-related mortality was noted for either i.t. or i.b. implantation when lower (1.0 x 10(5) or 1.0 x 10(4] tumor cell inocula were employed. Histological characteristics and growth patterns of tumors propagated employing the three implantation techniques were closely comparable for all three propagation methods and, in all instances, histological appearances of the tumors were representative of the current tumor cell lines from which they were derived. Approximately 30% of the lung tumors propagated i.t. grew in the chest wall and/or in the lung parenchyma as well as in the pleural space. In contrast, tumors propagated i.b. grew predominantly in the lung parenchyma. When five nonpulmonary human tumor cell lines (including U251 glioblastoma, LOX amelamontic melanoma, HT-29 colon adenocarcinoma, OVCAR 3 ovarian adenocarcinoma, and adriamycin-resistant MCF-7 breast adenocarcinoma) were propagated i.b. or i.t., there was considerable site-specific variability in tumor-related mortality depending on the tumor type. These data demonstrate that both the i.b. and i.t. models should be useful for the in vivo propagation and study of certain human pulmonary and nonpulmonary carcinomas as well as being advantageous for future studies of cancer biology and developmental therapeutics.

Feasibility of drug screening with panels of human tumor cell lines using a microculture tetrazolium assay.

For the past 30 years strategies for the preclinical discovery and development of potential anticancer agents have been based largely upon the testing of agents in mice bearing transplantable leukemias and solid tumors derived from a limited number of murine as well as human sources. The feasibility of implementing an alternate approach, namely combined in vitro/in vivo screening for selective cytotoxicity among panels of human tumor cell lines derived from a broad spectrum of human solid tumors is under investigation. A group of 30 cell lines acquired from a variety of sources and representing 8 lung cancer pathologies as well as 76 cell lines representing 10 other categories of human cancer (carcinomas of colon, breast, kidney, prostate, ovary, head and neck; glioma; leukemia; melanoma; and sarcoma) have exhibited acceptable growth characteristics and suitable colorimetric profiles in a single, standard culture medium. Measurements of in vitro growth in microculture wells by cell-mediated reduction of tetrazolium showed excellent correlation (0.89 less than r2 less than 0.98) with measurements of cellular protein in adherent cell line cultures as well as viable cell count in suspension cell line cultures (0.94 less than r2 less than 0.99). Since the microculture tetrazolium assay provides sensitive and reproducible indices of growth as well as drug sensitivity in individual cell lines over the course of multiple passages and several months' cultivation, it appears suitable for initial-stage in vitro drug screening.

Metastasis models for human tumors in athymic mice: useful models for drug development.

Although human tumor xenografts have been extensively used for preclinical evaluation of antitumor agents, most of this work has utilized subcutaneous or subrenal capsule assays based on change in tumor size. To obtain experimental models more reflective of the human clinical situations, we have developed several metastatic models that are based on and complement a panel of cell strains used in large-scale in vitro drug screening. One melanoma and four lung tumors produced metastatic lesions in the lung within 60 days following subcutaneous, intraperitoneal, or intrasplenic inoculation of BALB/C athymic nude mice. Several tumors also produced liver lesions, and one lung tumor strain showed metastasis to the brain. The metastatic lesions histologically resembled the tumors that grew at the inoculation site. In vitro and in vivo cell strains were rederived from the metastatic lesions. These systems may provide practical models for experimental drug and immunotherapeutic trials.

Characterization of infection and replication of Mason-Pfizer monkey virus in human cell cultures.

Human cells derived from both normal and neoplastic tissues can be infected by Mason-Pfizer monkey virus (MPMV) without accompanying cytopathology. Infection of cell cultures such as human rhabdomyosarcoma (A204) results in a persistenly infected cell line which can be subcultured over 30 sequential culture passages without significant change in phenotype properties according to reverse, transcriptase (RT), MPMV p27 antigen content, virus particle count and infectivity titre. Productive virus infections were established at relatively low virus particle (VP) input multiplicities (p.i.m.; about 0.06 VP/cell) In A204 cell cultures. At higher p.i.m. (about 600 to 6000 VP/cell) newly synthesized virus was detected within 4 days post infection. Although virus production was cumulative following primary infection, after subculture of infected cultures MPVM production was greater during active cell division. Using synchronization techniques, MPMV replication in persistently infected cultures was found to be cell cycle-dependent. The major internal antigen, p27, was synthesized in G2 and newly synthesized virus particles were released predominantly during mitosis and early G1. Colcemid arrest of cells during mitosis inhibited subsequent MPMV release. Consequently, production of extracellular virus depends upon the progression of cells through the mitotic stage. These data, which provided a basic understanding of the virus-host relationship that occurs in primate cells productively infected with MPMV, were used as a guideline for isolating MPMV-like viruses from experimentally and naturally infected Rhesus monkey.

Immunological characterization of mouse mammary tumor virus p10 and its presence in mammary tumors and sera of tumor-bearing mice.

Mouse mammary tumor virus (MMTV) p10 and gp52 were purified and used as radiolabeled antigens in sensitive radioimmunoassays. These radioimmunoassays were specific for MMTV proteins since detergent-disrupted MMTV from C3H/HeN, RIII, and GR/N mice gave complete competition, whereas C3H/HeNf liver extracts and other lysed retroviruses did not. Both gp52 and p10 are coded by the viral genome, since MMTV grown in a heterologous cell line (feline kidney cells) competed in these assays. Sera from mammary tumor-bearing mice and mammary tumors from C3H/HeN and C3H/HeNf mice competed in both the gp52 and the p10 assays. Although these radioimmunoassays detected predominantly group-specific antigenic determinants in C3H/HeN and C3H/HeNf tumor extracts, type specificity was also found with gp52. Absorption of the anti-MMTV serum with C3H/HeNf tumor extracts removed all antibodies directed against p10 and decreased the anti-gp52 titer approximately 30-fold. When this absorbed antiserum was used at limiting dilution in the gp52 radioimmunoassay, C3H/HeN tumor extracts gave complete competition, whereas no competition was found with C3H/HeNf tumor extracts.

Immunological characterization of the low-molecular-weight DNA binding protein of mouse mammary tumor virus.

p14, a low-molecular-weight MMTV protein previously identified as having DNA-binding properties and encoded by the gag region of the MMTV genome, was purified by affinity chromatography on DNA-sepharose. Immunological characterization of the purified protein showed that MMTV p14 shares no cross-reactivity with gp52, gp36 and p10, antigens associated with the MMTV envelope, nor with p27 antigen found in the virion core. Purified MMTV p14 did show cross-reactivity with purified intracytoplasmic A particles, supporting the concept that A particles are morphological precursors to MMTV cores. In addition, shared antigenic determinants between intracytoplasmic A particles and MMTV p27, p20 and p10 were demonstrated. MMTV p14 did not cross-react with the low-molecular-weight DNA-binding proteins of MuLV or of type-C or -D viruses of higher mammals.

Prevalence of murine mammary tumor virus antibody and antigens in normal and tumor-bearing feral mice.

Approximately 20% of normal male and female feral mice (Mus musculus) from areas with populations having either high [Lake Casitas (LC) and La Puente] or low (Bouquet Canyon) spontaneous lymphoma incidence expressed murine mammary tumor virus (MuMTV) gp52 in specific tissues. Sera from a low percentage (6%) of mice from the same trapping areas contained precipitating antibody specific for MuMTV. Although moderate to high levels of MuMTV gp52 were expressed in mammary tumor tissues of 3 of 7 LC mice and 3 of 3 (C57BL/10ScSn X LC)F1 mice, the same animals showed no detectable MuMTV-precipitating antibody. Neither MuMTV antibody nor tumor-associated MuMTV gp52 was defected in 10 LC mice bearing lymphomas or in 5 LC mice bearing hepatomas. Low levels of MuMTV gp52 expression and MuMTV antibody were also detected in subspecies of M. musculus and in the more distantly related species M. cervicolar. Compared with normal and tumor-bearing inbred mice of high (C3H/HeN) and low (C3H/HeN foster-nursed on NIH Swiss) mammary tumor strains, normal and tumor-bearing feral mice express MuMTV gp52 and MuMTV-precipitating antibodies at low frequency.

Natural distribution of squirrel monkey retrovirus proviral sequences in primate DNAs.

3H-labelled 70S RNA of squirrel monkey retrovirus (SMRV) hybridized to a high degree (greater than 52%) to the DNA of various tissues of two squirrel monkeys. Hybridization of the same probe to DNAs of other primates including New World monkeys (Woolly monkey, capuchin, owl monkey), Old World monkeys (rhesus, African green), apes (gibbon, chimpanzee), and human (A204 cells infected with MPMV) revealed no significant hybridization. Analysis of the kinetics of hybridization indicated that SMRV provirus was present in multiple copies in various squirrel monkey tissues (C0t 1/2 = 120 to 400) and in SMRV-infected A204 cells at a low number of copies (C0t 1/2 = 1500). These results demonstrate that SMRV is an endogenous virus of squirrel monkeys and the first isolated from a New World monkey.