Immunohistological detection of tumour growth fraction (Ki-67 antigen) in formalin-fixed and routinely processed tissues.
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Biomedical subjects
Publications and source records attributed to M H Becker.
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The monoclonal antibody Ki-67 reacts with a human nuclear cell proliferation-associated antigen that is expressed in all active parts of the cell cycle. Recently we have raised monoclonal antibodies, MIB 1-3, against recombinant parts of the Ki-67 antigen. These antibodies are true Ki-67 equivalents, as demonstrated by immunostaining of fresh specimens, biochemistry, and molecular biological techniques. Formalin-fixed, paraffin-embedded sections routinely processed for immunohistochemistry failed to stain for Ki-67 and MIB 2. Antibodies MIB 1 and MIB 3 labelled mitotic figures, while non-mitotic proliferating cells were negative under these conditions. However, when dewaxed microwave oven-processed paraffin sections of formalin-fixed tissues were used, MIB 1 and MIB 3 gave strong nuclear staining of those cells presumed to proliferate under a variety of normal and neoplastic conditions. Moreover, routine decalcification or depigmentation techniques did not alter the immunoreactivity of MIB 1 and MIB 3 with microwave-processed paraffin sections. This method is highly reproducible, easy to perform at low cost, and no additional technical skill is needed because after microwave treatment just routine immunohistochemical methods are used. Since we have successfully applied this new method to sections obtained from paraffin blocks stored for a long time (in one case more than 60 years), the assessment of cell kinetics through the detection of Ki-67 antigen is now possible on archival material collected in histopathology departments all over the world.
The immunohistochemical characteristics of the monoclonal antibody IND.64 are very similar to those of the monoclonal antibody Ki-67. The aim of this study was to further characterize this new antibody and to compare it with Ki-67 using immunobiochemical methods. Our results demonstrate that the similarity between the antibodies holds true even at the molecular level. Immunoblot analysis of IM-9-cell lysates with both antibodies showed a double band with apparent molecular weights of 395 kD and 345 kD, respectively. Competition ELISAs using a synthetic peptide derived from the thus far determined Ki-67 cDNA sequence as competitor, indicate that IND.64 may recognize the same epitope as Ki-67. The IND.64 epitope resides at least within a 20 amino acid sequence which also contains the Ki-67 epitope. Since IND.64 is of the IgG2b subclass, while Ki-67 is of the IgG1 subclass, the two antibodies may be useful for double immunostaining. In addition, IND.64 may help in determining the still unknown function of the antigen it recognizes.
This paper presents findings from the evaluation of a self-management education program based on self-regulation principles. Older men and women (N = 324) were randomly assigned to program and control groups. Outcomes were measured using the Sickness Impact Profile. Twelve months following baseline data collection, psychosocial functioning of program participants was significantly better than that of controls. Different program effects were noted when results were analyzed by participant gender.
AIM: To characterise a newly developed mouse monoclonal antibody JC1 which recognises a nuclear antigen present in proliferating cells in normal tissues and neoplastic lesions, and which is absent in resting cells. METHODS: The methodology was established using a representative range of frozen sections from normal tissues and from certain tumours which were immunostained with antibodies Ki67 and JC1. The molecular weight of the antigen recognised by JC1 was obtained by western blot analysis and this was compared with that of Ki67. IM-9 cell lysates containing Ki67 derived plasmids were also tested with JC1 antibody. RESULTS: Biochemical investigation indicated that the antigen recognised by JC1 gives two molecular weight bands of 212 and 123 kilodaltons, which is distinct from the well characterised anti-proliferation monoclonal antibody Ki67 (395-345 kilodaltons). In addition recombinant Ki67 protein is not recognised by JC1. Immunohistological reactivity was seen in areas known to contain numerous proliferating cells such as lymphoid germinal centres, splenic white matter, cortical thymocytes and undifferentiated spermatogonia. In tumours many cells from adenocarcinomas, oat cell carcinomas, squamous cell carcinomas of lung, and seminomas were labelled by JC1 with a distribution and proportion similar to that seen with Ki67. In normal tissues the only apparent difference was in testis where JC1 stained a considerably greater number of cells than Ki67. In all cases studied the new antibody showed nuclear reactivity only. JC1 did not show any cytoplasmic crossreactivity with squamous cells as is frequently seen with Ki67. CONCLUSION: Antibody JC1, which recognises a nuclear antigen present in proliferating cells, should provide a useful adjunct to Ki67 as a marker of proliferation especially in those cases such as squamous cell carcinomas where a Ki67 index cannot be determined.
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In 1987 the Centers for Disease Control published a Universal Precautions Policy establishing blood and body fluid procedures to be used consistently with all patients. An important and unequivocal Universal Precautions Policy recommendation with regard to avoidance of needlestick injuries is that needles should never be recapped. We examined the recapping-related attitudes and behaviors of physicians and nurses at four large teaching hospitals with patients with acquired immunodeficiency syndrome and with Universal Precautions Policy in-service training programs. Compliance was found to be considerably less than optimal. According to unannounced needle counts in disposal boxes, the percentage of recapped needles was always greater than 25% and exceeded 50% in four instances. Recapping was related to inadequate knowledge, concerns about personal risk, forgetfulness, being "too busy" to follow the Universal Precautions Policy, and the misperception that recapping is a way to avoid needlestick injury. Strategies are suggested to improve and supplement traditional in-service education.
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To investigate the effects of an educational program on the compliance-enhancing behavior of pediatricians and the subsequent regimen adherence of their patients, it was necessary to enlist the participation of a large number of pediatric practitioners. The physicians had to be motivated to attend two evenings of tutorial training; to enroll their patients in the project; and to complete detailed study documents. Ultimately, 90 of the 97 pediatricians practicing in the community took part. The following factors are felt to have contributed to the widespread participation of the pediatricians: 1) the subject was interesting and of relevance to clinical practice; 2) the study was designed so as not to interfere with office routine; 3) major demands were not made on the patients; 4) due consideration was given to the value of the physicians' participation; and 5) communication between the investigators and the practitioners was facilitated by a liaison pediatrician.
The Health Belief Model (HBM) has been applied to a variety of health conditions: most are less threatening and require less complex responses than those arising in the case of AIDS. The utility of the HBM in understanding preventive behaviors in AIDS is examined in a cohort of homosexual men at two different time points. Longitudinal analyses estimated the relationship of indices assessing susceptibility, severity, benefits, and barriers, as well as sociodemographic factors, to sexual behavior across the next 18 months. Analyses further defined these associations in specific subgroups of participants, such as those initially at lower or higher risk. In general, measures of severity and socioeconomic advantage had the most consistently beneficial effect on various measures of behavior. Little or no beneficial effect was observed for other components of the HBM, notably a measure of perceived susceptibility. These analyses suggest that the special features of AIDS may require development of more adequate theoretical frameworks.
The campomelic syndrome is characterized by dwarfism, craniofacial anomalies, bowing of the tibiae and femora, cutaneous dimpling overlying the tibial bend, respiratory distress, and early death. Otolaryngologic manifestations include flat facies with a broad nasal bridge, low-set ears, cleft palate, mandibular hypoplasia, and tracheobronchial malacia. The underlying pathologic feature appears to be disturbance in cartilage growth involving the affected bones and the respiratory tract cartilage. The cause is unknown. We report clinical and histopathologic features in two cases of this syndrome. The endochondral layer of the otic capsule contained no cartilage cells. The cochlea was short and flattened, presenting a scala communis. The vestibule and the canals were deformed by bone invasion. Defective endochondral ossification of the petrooccipital synchondroses possibly explains the shortened skull base seen in this syndrome. The tracheobronchial malacia significantly contributes to respiratory distress and neonatal death.
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Negative attitudes toward obese patients by health professionals have been attributed to termination of weight reducing attempts. This study measured attitudes concerning the obese held by professional participants in a continuing education conference on causes and treatment of obesity. Substantial variation was found on items comprising obesity-related belief dimensions: (a) Disparaging image of the obese; (b) causes of obesity; and (c) ways to lose weight. Evidence is presented for the association among measures of these dimensions. While background and educational characteristics and conditions of practice were not related to such attitudes, the professional's personal experience with successful weight reduction was the best predictor of favorable attitudes.
A field experiment was designed to test the effects on weight change in obese children of (1) communications varying in threat and (2) health beliefs of the child's mother, in terms of perceived vulnerability to health threats, the efficacy of taking actions, and barriers to acting. Treatments involved messages containing more or less threatening material regarding obesity, plus a control (no communication) condition. Beliefs were assessed by means of personal interviews. All mothers and children received dietary counseling and were scheduled for four follow-up visits, which included weighing. The 182 participants were from low-income areas served by a large hospital pediatric clinic. High-threat messages yielded the most consistent weight loss, followed by low threat and control. In addition, both general beliefs concerning health and specific beliefs about obesity and dieting predicted weight loss. Both messages and mothers' beliefs acted together on keeping appointments and making efforts to comply. Results are discussed in terms of the effects of threats and health belief-behavior linkages.
Health services utilization may be influenced by the structure of the health system and the behavior of health professionals as well as by the actions of individual patients. This research examines the responses of obstetricians toward women seeking abortion. The population for this study includes all obstetrician-gynecologists with any private practice in Maryland during 1975 (473). Each responding physician (443) was presented with a case history vignette describing, in a telephone interview, a woman who is pregnant and considering an abortion. The sociodemographic characteristics of the woman were systematically varied to determine effects of patient attributes on physicians' patient management decisions. Decisions to refer the patient or to participate personally in her care were found to be associated most strongly with the patients' financial resources. Three hundred and twelve obstetricians returned a mail questionnaire, probing their own attitudes and characteristics. Physicians' liberal or conservative attitudes toward expansion of reproductive health care services and their level of disturbance by the abortion procedure were also influential in these patient management decisions. Simultaneous examination of both patient and physician characteristics indicated that the former had the greater weight in accounting for referral decisions.