ABSTRACT
Antigen detection with indirect immunohistochemical methods is hampered by high background staining if the primary antibody is from the same species as the examined tissue. This high background can be eliminated in unfixed cryostat sections of mouse skeletal muscle by boiling sections in PBS, and several proteins including even the low abundant dystrophin protein can then be easily detected with murine monoclonal antibodies. However, not all antigens withstand the boiling procedure. Immunoreactivity of some of these antigens can be restored by subsequent washing in Triton X-100, whereas immunoreactivity of other proteins is not restored by this detergent treatment. When such thermolabile proteins are labeled with polyclonal primary antibodies followed by dichlorotriazinylaminofluorescein-conjugated secondary antibodies and boiled, the fluorescence signal persists, and sections can then be processed with a monoclonal antibody for double immunostaining of a protein unaffected by boiling. This stability of certain fluorochromes on heating can also be exploited for double immunofluorescence labeling of two different thermostable proteins with murine monoclonal antibodies as well as for combination with Y-chromosome fluorescence in situ hybridization. Our method should extend the range of monoclonal antibodies applicable to tissues derived from the same species as the monoclonal antibodies.
Subject(s)
Antibodies, Monoclonal , Antigens/metabolism , Muscle, Skeletal/metabolism , Animals , Antigens/immunology , Buffers , Dystrophin/immunology , Dystrophin/metabolism , Frozen Sections , Hot Temperature , Immunohistochemistry , In Situ Hybridization, Fluorescence , Male , Mice , Mice, Inbred mdx , Phosphates , Sodium Chloride , Solutions , Y ChromosomeABSTRACT
Relapse prevention plays an important role in the treatment of alcoholics. Positive predictors are high self-efficacy and confidence in high-risk situations, positive outcome expectancies, sufficient coping-skills and low craving. We examined 64 primary German alcoholics. Each person met the criteria of alcohol dependence of DSM-IV and ICD-10. Subjects were included in an in-patient 4-step relapse prevention program. This manual-guided group psychotherapy is based on cognitive-behavioral approach focused on high-risk situations, phases of relapse, stress/relax and resources. Craving before and after treatment did not differ, so that using this kind of relapse prevention training might be suitable in both in- and out-patient programs. High values for insight, optimism, self-efficacy and confidence in high-risk situations might enhance individual coping-skills to balance patient's lifestyle focussing on an abstinent way of life.
ABSTRACT
Relapse prevention plays an important role in the treatment of alcoholics. Positive predictors are high self-efficacy and confidence in high-risk situations, positive outcome expectancies, sufficient coping-skills and low craving. We examined 64 primary German alcoholics. Each person met the criteria of alcohol dependence of DSM-IV and ICD-10. Subjects were included in an in-patient 4-step relapse prevention program. This manual-guided group psychotherapy is based on cognitive-behavioral approach focused on high-risk situations, phases of relapse, stress/relax and resources. Craving before and after treatment did not differ, so that using this kind of relapse prevention training might be suitable in both in- and out-patient programs. High values for insight, optimism, self-efficacy and confidence in high-risk situations might enhance individual coping-skills to balance patient's lifestyle focussing on an abstinent way of life.
