Immunochemistry of apamin—bee venom neurotoxin—I. Radioimmunoassay with apamin and its derivatives

Antibodies against apamin, a neurotoxic polypeptide from bee venom were raised in rabbits by immunization with apamin or apamin-BSA conjugates. ³H-apamin or ¹²⁵I-apamin were used in radioimmunoassay with anti-apamin for the detection of the apamin antigenic site. The inhibitory activity toward the labelled apamin-anti-apamin binding was maximal with unlabelled apamin and decreased in the range: apamin > Cys¹, Lys⁴-disuccinilated apamin > Cys¹, Lys⁴-diacetylated apamin > Cys¹, Lys₄-diacetylated apamin with carboxymethylated His¹⁸. Dipyrimidyl-Orn¹³,Orn¹⁴-apamin derivative almost had no inhibitory activity on labelled apamin binding emphasizing that Arg¹³, Arg¹⁴ are the most essential for the apamin topographic antigenic site.     

A monoclonal antibody (SJ-9A4) to P24 present on common alls, neuroblastomas and platelets - I. Characterization and development of a unique radioimmunometric assay

We report the development and characterization of SJ-9A4, a monoclonal antibody (MoAb) produced against common acute lymphoblastic leukemia (C-ALL) cell lines. SJ-9A4 reacted with C-ALL, B-cell chronic lymphocytic leukemia (B-CLL), platelets and C-ALL neuroblastoma (NB) and the K562 cell lines. It had no significant reactivity with erythrocytes, granulocytes, circulating T or B lymphocytes, monocytes, granulocytic cell lines or a Ewing's sarcoma cell line. SJ-9A4 was shown to recognize the same region as two other MoAb to the p24 antigen, BA-2 and DU-ALL-1, as demonstrated by their ability to inhibit the binding of labeled SJ-9A4 to NALM-1 and NB cells. Other MoAb: J5, PI 153/3 and monoclonal anti-HLA-DR antibodies gave no inhibition. A solid phase indirect radioimmunometric assay (IRA) was developed which enabled the detection of P24 from C-ALL cells, utilizing its ability to bind the Ricinus communis agglutinin (RCA1) or wheat germ agglutinin (WGA) and SJ-9A4 simultaneously. When BA-2 and DU-ALL-1 were used in place of SJ-9A4, similar IRA results were obtained. Using the RCA1/SJ-9A4-IRA, P24 from as few as 1.6 X 10(4) cells of a C-ALL cell line could be detected; however, similar extracts of NB cell lines were negative despite high levels of SJ-9A4 binding to intact cells. The presence of P24 in NB extracts was demonstrated by (1) preincubation of NB extracts with SJ-9A4 which blocked MoAb binding to P24 and (2) immunoadsorption of P24 from solubilized membranes of 35S-methionine (met) labeled NB cells. Treatment of NB cells with neuraminidase did not result in IRA binding when either RCA1 or WGA were used as the solid phase lectin indicating that the differences in lectin affinity are not due to over sialation of NB membrane glycoproteins. These findings demonstrate a difference in the glycosylation of P24 from C-ALL and NB cells.     

Activation of hepatic guanylate cyclase by nitrosyl-heme complexes: Comparison of unpurified and partially purified enzyme

The objective of this study was to evaluate the effects of several agents on activation of both unpurified and partially purified hepatic soluble guanylate cyclase by performed NO (nitric oxide or nitrosyl)-heme complexes. Guanylate cyclase was activated by NO complexes of the heme compounds, hematin, hemoglobin, myoglobin, catalase and cytochrome c, and also by the reaction product of NO and ferredoxin, a non-heme, iron sulfur electron transfer protein. NO-lipoxygenase, which contains non-heme iron, did not activate guanylate cyclase. NO-heme complexes activated unpurified enzyme almost equally well in the presence of either Mg²⁺ or Mn²⁺. However, activation of purified (350- to 750-fold) guanylate cyclase was markedly greater with Mg²⁺ than with Mn²⁺. At concentrations that did not alter basal enzymatic activity, Ca²⁺ markedly inhibited guanylate cyclase activation in the presence of Mg²⁺ but not of Mn²⁺. Hemoproteins inhibited activation of unpurified and purified enzyme by NO-heme complexes, and increasing the concentrations of the latter overcame the inhibition. Gel filtration studies indicated that uncomplexed and NO-complexed hematin bind to common or adjacent sites on guanylate cyclase. Whereas dl-dithiothreitol enhanced activation, ferricyanide, cystine, o-iodosobenzoic acid and ethacrynic acid inhibited activation of guanylate cyclase by NO-heme complexes. The data indicate that the effects of these diverse agents on guanylate cyclase activation by preformed NO-heme are similar to their effects on enzyme activation by NO and nitroso compounds, both of which readily form NO-heme complexes. Therefore, the effects of these diverse agents may be on guanylate cyclase rather than on NO-heme formation.     

Eliminating a severely retarded blind adolescent's tantrums using mild behavioral interruption: A case study

A blind 19 yr old severely retarded man was referred for behavior therapy because of violent and disruptive tantrums. Previous behavioral strategies had failed for various reasons. A very mild, brief, vestibular oriented physical procedure was designed to provide a low level disruptive effect. Intervention consisted of loud teacher demands to stop and work appropriately as well as guiding him through one 360° turn while standing. The data demonstrated rapid and long term success in eliminating the client's tantrums. The results were interpreted in terms of a behavioral interruptor of a chained sequence allowing refocusing of client attention and increased levels of reinforcement.     

Studies using releasing factors in Klinefelter's Syndrome: The responses of LH and FSH to luteinizing hormone-releasing hormone and of prolactin and TSH to thyrotropin-releasing hormone before and after testosterone

(1) During an investigation of their pituitary function, the TSH prolactin (PRL) and gonadotropin, (LH, FSH) response to TRH was measured in four patients with documented euthyroid Klinefelter's syndrome. (2) In all four patients, the serum testosterone was low, the gonadotropins were elevated and thyroid antibodies undetectable. (3) The identical study was repeated after each patient had received intramuscular testosterone cypionate 200 mg bi-weekly for three doses. This raised the serum testosterone at least four-fold. (4) Basal TSH was normal in all four patients but its response to TRH was blunted (mean peak TSH 9.6 μU/ml), compared with normal controls (mean peak TSH 18 μU/ml). Following testosterone this response to TRH was further suppressed to a mean peak of 3.4 μU/ml TSH. (5) Basal PRL levels were also normal and the mean peak response to TRH, 28 ng/ml. When repeated after testosterone, TRH produced a mean peak PRL of 58 ng/ml. (6) LH and FSH, both elevated basally, did not change with TRH administration, but both were stimulated by LHRH 100 μg i.v., LH briskly, FSH more variably. (7) FSH was suppressed sharply but LH only partially with testosterone therapy. (8) Our studies indicate that the TSH response to TRH in Klinefelter's syndrome is blunted and further suppressed with short term testosterone replacement. The PRL response to TRH under these same circumstances, however, is enhanced. Previous reports of abnormal LH feedback control are confirmed. (9) Klinefelter's syndrome is associated with subtle abnormalities in hypothalamic pituitary regulation not expected in typical primary hypogonadism.     

Increased excretion of N-carbamoyl compounds in patients with urea cycle defects.

Increased urinary levels of N-carbamoyl-beta-alanine, and also, on occasions, of N-carbamoylaspartate, were observed in patients with ornithine carbamoyl-transferase (EC 2.1.3.3) deficiency, argininosuccinate synthetase (EC 6.3.4.5) deficiency and argininosuccinate lyase (EC 4.3.2.1)deficiency, but not in a patient with carbamoylphosphate synthase deficiency. The relevance of these findings to the diagnosis of urea cycle defects is discussed.     

A methodological critique of Bandura's self-efficacy theory of behavior change

This brief theoretical note identifies a major methodological problem in Bandura's argument in favor of self-efficacy theory and provides two methods of controlling for it. His data could likely be accounted for by social contingencies operating within his highly structured behavioral approach situation.     

An unusual case of pleural, epididymal and sternoclavicular tuberculosis

A patient with pleural, epididymal and sternoclavicular tuberculosis is described in whom the history suggests that primary infection occurred near the time when he was involved in a fight. Epididymal and sternoclavicular lesions represent infection of loci minoris resistentiae secondary to traumatic injuries sustained in the fight and provide evidence for the concept of tuberculous bacillaemia accompanying primary infection.     

Effects of task difficulty on EEG measures of cerebral engagement

EEGs were recorded while subjects constructed 14 block designs of graded complexity, and also while reading and writing. Two measures of design difficulty were used: time to complete each construction and Ss' subjective ranking. For each task, average alpha power for each lead and R/L ratios (C₄/C₃, P₄/P₃) were calculated.Within-subject and group analyses both showed higher alpha power and higher R/L ratios with increasing difficulty. There were important differences between subjects: some showed significant increases in alpha power only on the left, some only on the right, some bilateral. Regardless of difficulty however, the R/L alpha ratios were significantly lower for block design than for writing, as in our previous studies.