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R D Lane

Publications and source records attributed to R D Lane.

68 records · Page 4Linked to original sources
PubMed1985

A short-duration polyethylene glycol fusion technique for increasing production of monoclonal antibody-secreting hybridomas.

In this study the exposure period of the lymphocyte-myeloma cell mixture to the fusogen was evaluated for its influence upon the yield of total hybridoma colonies and those which secreted monoclonal antibodies. Sp2/0 and FOX-NY myeloma cells were fused for varying periods with murine splenic lymphocytes immunized with sheep red blood cells. The optimal fusion period consisted of adding the fusogen (5.0 ml Kodak 1450 PEG, 0.5 ml dimethylsulfoxide, and 4.5 ml of phosphate-buffered saline, pH 7.0) to the cell mixture over a 45 s period at 37 degrees C. The fusion process was stopped by gradually diluting the mixture in 50 ml of RPMI-1640. After 10 min, the cells were centrifuged, resuspended in selective medium with feeder macrophages and cultured. In comparison to common, longer fusion techniques, this procedure produces approximately a 5-fold increase in the number of hybrids produced when using the Sp2/0 cells and a 30-fold increase in the number of hybrids produced when using the FOX-NY cells as the fusion partner. In both cases, virtually all the wells contain monoclonal antibody-secreting hybridoma colonies. This high efficiency fusion technique can be used most advantageously to produce monoclonal antibodies against weak immunogens or to reduce the time needed for immunization with stronger immunogens.

Animals↗
PubMed1985

Subcellular localization of bovine heart calcium-dependent protease inhibitor.

Four monoclonal antibodies for a calcium-dependent protease inhibitor protein were produced by fusing Sp2/0 myeloma cells with spleen cells from a Balb/C mouse immunized with purified bovine heart inhibitor. Each of the monoclonal antibodies was highly specific for the inhibitory protein as revealed by electro-blot analysis. The antibodies recognized different antigenic sites on CNBr peptides prepared from the purified inhibitor protein. Immunofluorescent microscopy of sections from bovine heart ventricles treated with each of the antibodies demonstrated the same fluorescent pattern. Fluorescence was observed at or near the sarcolemma of the myocytes, and along the Z-discs of relaxed myofibrils within the myocytes. Contracted myofibrils did not appear to bind antibody. Immunostaining of glycerinated relaxed cardiac myofibrils revealed staining at the Z-discs. One of the antibodies could also stain the Z-disc region of bovine skeletal muscle myofibrils.

Animals↗
PubMed1985

Assessment of tardive dyskinesia using the Abnormal Involuntary Movement Scale.

Over a 10-month period, 33 patients with tardive dyskinesia (TD) were evaluated with the Abnormal Involuntary Movement Scale (AIMS) simultaneously and independently by two experienced and two inexperienced raters. The experienced raters generally had higher levels of agreement and their scores were more consistent over time. It is concluded that experience with TD influences AIMS inter-rater reliability and that it is useful to differentiate TD movements into the dimensions of quality, frequency, and amplitude, dimensions not currently used in the AIMS. The usefulness and difficulty of developing more specific guidelines for AIMS ratings are discussed.

Ambulatory Care↗
PubMed1984

Comparison of polyethylene glycols as fusogens for producing lymphocyte-myeloma hybrids.

In order to improve the yield of hybridomas for monoclonal antibody production, 8 different sources and molecular weights of polyethylene glycol (PEG) were compared as fusing agents. Sp2/0 myeloma cells were fused with murine splenic lymphocytes immunized with sheep red blood cells. The Kodak 1450 PEG produced the maximum number of hybridomas. The optimal technique consisted of slowly adding 1 ml of freshly prepared fusogen (5 g Kodak 1450 PEG, 0.5 ml dimethylsulfoxide, and 5 ml of phosphate-buffered saline, pH 7.0) to the cells over a 1 min period, incubating the mixture at 37 degrees C for 90 s, then gradually diluting the mixture in 50 ml of Hanks' buffered salt solution. After 10 min, the cells are centrifuged, resuspended in selective medium with feeder macrophages and cultured. This procedure routinely produces between 600-3,000 hybridomas per fusion.

Animals↗
PubMed1984

Localization of a macrophage chemokinetic factor on neoplastic cells.

A rabbit antiserum was produced against a high-molecular-weight fraction (360,000) of murine Lewis lung carcinoma (LLC)-conditioned media. This fraction contained a factor that increased the rate of random migration (chemokinesis) of activated macrophages (M phi) in male C57BL/6J mice. The specificity of the antiserum was demonstrated by the ability of the antiserum to absorb the chemokinetic activity from tumor-conditioned media. Immunofluorescence studies demonstrated cell surface localization of antigenetically similar material on the LLC, Ehrlich ascites cells, MCA9/14 and MCA64/8 mouse fibrosarcomas, and MBT-2 mouse bladder carcinoma. The antiserum further indicated the presence of the chemokinetic factor (CKF) on the surface of peritoneal M phi previously exposed to tumor media. The CKF was observed on approximately 90% of the Corynebacterium parvum-activated M phi and of the M phi activated by maleic anhydride-divinyl ether copolymer (fraction 2), on 10% of the oyster glycogen-elicited M phi, and on 0% of the unstimulated M phi. these data support the concept that the CKF is a common surface marker of neoplastic cells and that it is bound by activated tumoricidal M phi.

Animals↗
PubMed1982

Characterization of a macrophage chemokinetic factor in tumor cell culture media.

Media conditioned by tumor cells were studied for the presence of factor(s) that increase the rate of random migration (chemokinesis) of Corynebacterium parvum-activated macrophages. A capillary tube assay was developed and utilized to expediently monitor the chemokinetic activity of macrophages incubated in whole and fractionated media. Media conditioned by six different syngeneic and allogeneic mouse tumor cell lines demonstrated significantly higher chemokinetic activity than unconditioned or normal fibroblast conditioned media. The chemokinetically active component of the Lewis Lung conditioned media was found to be a trypsin sensitive, heat stable, high molecular weight (300,000-480,000 dalton range) factor that had no chemotactic (directional migration) activity. Pyran-activated macrophages also responded chemokinetically to the Lewis Lung factor while oyster glycogen and thioglycolate-elicited macrophages did not. The similarity and differences between the physical properties of the chemokinetic factor, other migration stimulating factors, and tumor-associated proteins are discussed.

Animals↗
PubMedDate not supplied

Sociodemographic correlates of alexithymia.

The association between alexithymia and sociodemographic variables is not well understood. Previous studies using the current 20-item and previous 26-item versions of the Toronto Alexithymia Scale (TAS) have shown inconsistent associations with age, sex, socioeconomic status, and years of education. We tested 380 subjects from the community stratified equally across sex, five age groups, and three socioeconomic classes. In addition to the TAS-20, we also administered the Levels of Emotional Awareness Scale (LEAS), a behavioral measure of the ability to be aware of and represent emotions in words, a core component of alexithymia. The TAS-20 and LEAS were each correlated with age, sex, socioeconomic status (SES), and years of education (P < .01) in the same direction and to approximately the same degree. Alexithymia (or low emotional awareness) is associated with older age, male sex, lower SES, and fewer years of education. The TAS-20 and LEAS are only slightly correlated (r = -.19, n = 380, P < .001), but their correlation is largely accounted for by their shared variance with these demographic variables. The convergence of findings with these two quite different measures and the nature of their overlap support the validity of these associations between alexithymia and sociodemographic variables.

Adolescent↗
PubMedDate not supplied

Projection status of calbindin- and parvalbumin-immunoreactive neurons in the superficial layers of the rat's superior colliculus.

Immunocytochemistry and retrograde labeling were used to define the thalamic projections of calbindin- and parvalbumin-containing cells in superficial layers of the rat's superior colliculus (SC). Quantitative analysis revealed that 90.8 +/- 2.2% (mean +/- standard deviation) of the calbindin-immunoreactive neurons in the stratum griseum superficiale (SGS) projected to the dorsal lateral geniculate nucleus (LGNd) and that 91.3 +/- 4.3% of calbindin-immunoreactive neurons in the stratum opticum (SO) projected to the lateral posterior nucleus (LP). In contrast, only 17.3 +/- 2.5% of parvalbumin-immunoreactive neurons in the SGS were found to project to the LGNd and 16.5 +/- 3.1% of the parvalbumin-immunoreactive SO cells were retrogradely labeled after LP injections. Few of the parvalbumin-immunoreactive neurons in either the SGS (7.2 +/- 2.5%) or the SO (9.2 +/- 2.5%) were GABA positive. The retrograde-labeling results suggest that parvalbumin-immunoreactive neurons in the rat's SO and SGS may either be primarily interneurons or have descending projections, while calbindin-containing cells are primarily thalamic projection neurons. These results are consistent with data from other rodents, but almost exactly the opposite of data that have been reported for the cat for these same populations of SC projection neurons. Such interspecies differences raise questions regarding the functional importance of expressing one calcium-binding protein versus another in a specific neuronal population.

Animals↗
PubMedDate not supplied

Induction of lateralized sympathetic input to the heart by the CNS during emotional arousal: a possible neurophysiologic trigger of sudden cardiac death.

A neurophysiologic mechanism for stress-induced cardiac arrhythmias is proposed based on the integration of two bodies of research that have until now developed independently: the role of hemispheric specialization in the mediation of emotional arousal and the role of a lateralized imbalance in sympathetic input to the heart in cardiac arrhythmogenesis. The specific hypothesis is that individuals who manifest more lateralized frontal lobe activity during emotional arousal may concomitantly generate more lateralized sympathetic input to the heart and be at increased risk for fatal cardiac arrhythmias. The theoretical background for the hypothesis is presented, empirical support for the hypothesis is reviewed, and implications for empirical research are discussed.

Arousal↗
PubMedDate not supplied

Impaired verbal and nonverbal emotion recognition in alexithymia.

Although clinical observations suggest that alexithymic individuals have a deficit in their ability to recognize emotional stimuli and that this deficit is not simply due to a problem in verbal labeling, these two hypotheses have not been empirically confirmed. Three hundred eighty participants in a community survey without current or past histories of psychiatric disorder completed two independent measures of alexithymia [the Levels of Emotional Awareness Scale (LEAS) and the Toronto Alexithymia Scale (TAS-20)] and the Perception of Affect Task (PAT), a 140-item measure of the ability to match emotion stimuli. The PAT includes four subtasks that require the subject to match verbal or nonverbal emotion stimuli with verbal or nonverbal emotion responses. The subtasks include matching sentences and words (verbal-verbal), faces and words (nonverbal-verbal), sentences and faces (verbal-nonverbal), and faces and photographs of scenes (nonverbal-nonverbal). Across the entire sample, higher (alexithymic) TAS-20 and lower LEAS scores were both correlated with lower accuracy rates on each of the subtasks of the PAT (p < .001), accounting for 10.5% and 18.4% of the variance, respectively. Fifty-one subjects met TAS-20 criteria for alexithymia. Alexithymic individuals scored lower than other subjects on purely nonverbal matching, purely verbal matching, and mixed verbal-nonverbal matching (all p < .001). These results suggest that alexithymia is associated with impaired verbal and nonverbal recognition of emotion stimuli and that the hallmark of alexithymia, a difficulty in putting emotion into words, may be a marker of a more general impairment in the capacity for emotion information processing.

Adolescent↗
PubMedDate not supplied

Pervasive emotion recognition deficit common to alexithymia and the repressive coping style.

OBJECTIVE: Previous research has demonstrated a deficit in the ability to recognize emotions in alexithymic individuals. The repressive coping style is thought to preferentially impair the detection of unpleasant compared with pleasant emotions, and the degree of deficit is typically thought to be less severe than in alexithymia. We compared emotion recognition ability in both individuals with alexithymia and those with the repressive coping style. METHODS: Three hundred seventy-nine subjects completed the 20-item Toronto Alexithymia Scale, the Levels of Emotional Awareness Scale, the Marlowe-Crowne Scale (a measure of repressive defensiveness), the Bendig Short Form of the Taylor Manifest Anxiety Scale, and the Perception of Affect Task. The Perception of Affect Task consists of four 35-item emotion recognition subtasks: matching sentences and words, faces and words, sentences and faces, and faces and photographs of scenes. The stimuli in each subtask consist of seven emotions (happiness, sadness, anger, fear, disgust, surprise, and neutral) depicted five times each. Recognition accuracy results were collapsed across subtasks within each emotion category. RESULTS: Highly alexithymic subjects (for all, p<.01) and those with low emotional awareness (for all, p<.001) were consistently less accurate in emotion recognition in all seven categories. Highly defensive subjects (including repressors) were less accurate in the detection of anger, sadness, fear, and happiness (for all, p<.05). Furthermore, scores on the Levels of Emotional Awareness Scale accounted for significantly more variance in performance on the Perception of Affect Task than scores on the Marlowe-Crowne Scale (p<.01). CONCLUSIONS: The results indicate that alexithymia and the repressive coping style are each associated with impairments in the recognition of both pleasant and unpleasant emotions and that the two styles of emotional self-regulation differ more in the magnitude than in the quality of these impairments.

Adaptation, Psychological↗
PubMedDate not supplied

The neuropsychophysiology of emotion.

The principles of hierarchical organization and progressive increase of differentiation and integration within a hierarchy provide a framework for understanding how the subjective experience of emotion, brain mechanisms mediating emotion and autonomic activation during emotional arousal are organized, and how the three domains are interrelated. Supporting evidence from empirical studies of three types, pairing one domain with each of the other two, are reviewed. This integrated model may bring coherence to the study of emotion, and facilitate an improved understanding of the role of emotion in systemic medical disorders.

Adult↗