The immune response of inbred mouse strains to DNP-BGG

The immune response of six inbred mouse strains (SJL, A, C57BL/6, CBA, BALB/c, and DBA/1) to DNP₅₆BGG was tested under three separate immunization schedules: 1 Μg DNP-BGG in 1 mg Al(OH)₃ adjuvant, 50 Μg DNP-BGG in 1 mg A1(OH)₃ adjuvant, and 1 Μg DNP-BGG in complete Freund's adjuvant. Individual serum samples were titered using a modified Farr assay. It was found that the first schedule allowed classification of the mice into responder (SJL, A) and nonresponder (C57BL/6, CBA, BALB/ c, DBA/1) strains. The second schedule produced quantitative as well as qualitative differences among the strains and allowed classification of the mice into higher-responder (SJL, A), intermediate-responder (C57BL/6, CBA, BALB/c), and low-responder (DBA/1) categories. When complete Freund's adjuvant was used in the third schedule, the differences among strains became insignificant. The sera from each strain were pooled and assayed for relative antibody affinity and IgM content. Both of these parameters were dependent largely on the dose of antigen and type of adjuvant used, rather than on the particular mouse strain being studied. The mechanism of adjuvant action, and possible cell interactions in the genetic control of the immune response, are discussed.     

Phototropism in a red alga, Griffithsia pacifica

In the red alga, Griffithsia pacifica, shoot portions of a plantare positively phototropic and rhizoids are negatively phototropic.We have studied the phototropic response of rhizoids which elongateby tip growth. For 45 min after the beginning of unilateralillumination a rhizoid grows straight, then phototropic curvaturebegins and continues rapidly until the rhizoid is growing awayfrom the light. Curvature is 70–80% complete after 3 hr.If the unilateral stimulus is given for a short time (15 min),curvature again begins at 45 min. However, within an additional30–45 min the rhizoid stops growing away from the lightand wanders back towards its original direction of growth. Phototropismis elicited by light of wavelengths from 350 nm to 500 nm; inlight of wavelengths above 550 nm, little, if any, responseoccurs. ¹Present address: Division of Natural Sciences, University ofCalifornia, Santa Cruz, California 95064, U.S.A. (Received December 10, 1976; )     

The role of citrate in lipid synthesis in developing soybean cotyledons

The role of citrate and the citrate cleavage enzyme in lipidsynthesis in developing soybean cotyledons (Glycine max L. Merr.var. Harosoy 63) was investigated. The activity of the enzymewas inhibited by (—) hydroxycitrate, which is a specificinhibitor of citrate cleavage by this enzyme. Incorporationof label from citrate-1-¹⁴C and -5-¹⁴C indicated that the citratemolecule is cleaved between carbons 3 and 4. Acetyl CoA-¹⁴Cand oxaloacetate-¹⁴C phenylhydrazone were isolated as productsof the citrate cleavage reaction. The production of oxaloacetate-¹⁴C-phenylhdrazonefrom citrate-6-¹⁴C was carried out using a nucleotide free enzymepreparation and did not require the addition of ATP or CoA.Therefore it would appear that the citrate cleavage reactionis not CoA dependent in developing soybean seeds. Incorporationof pyruvate-2-¹⁴C into the crude lipid fraction was shown torequire both the particulate and soluble fractions. Apparentlyin soybeans, as in animal systems, pyruvate is oxidized by thepyruvate dehydrogenase complex and the acetyl CoA formed condenseswith oxaloacetate to produce citrate in the mitochondria. Citrateis then transported out of the mitochondria to the cytosol whereit is cleaved to form acetyl CoA for lipid synthesis. ¹ Cooperative investigations of the Agricultural Research Service,U.S. Department of Agriculture, and Illinois Agricultural ExperimentStation. ² This research represents partial fulfillment of the Ph. D.requirements of Daniel R. Nelson. Presently at Monsanto AgriculturalProducts Co., St. Louis, MO 63141, U.S.A. (Received January 12, 1977; )     

Subtypes of Depression—Diagnosis and Medical Management

Depression is both a common and a greatly undertreated illness in the United States today. The focus of this review is a definition of the characteristics of four subtypes of depression which appear to be differentially sensitive to four different classes of medications. The tricyclic antidepressants should be used for patients with unipolar depression and vegetative symptoms. Lithium appears to be most effective for bipolar depressives. The monoamine oxidase (MAO) inhibitors are best used for patients with atypical depression. Antipsychotic medications appear to be useful for depressed patients with psychotic symptoms or agitation. Recent pharmacokinetic and biochemical data, including serum lithium levels, plasma tricyclic levels, and the predictive ability of pretreatment urinary 3-methoxy-4-hydroxyphenylglycol (MHPG) levels are also reviewed.     

Familial Mediterranean Fever: Recent Advances in Pathogenesis and Management

The success of colchicine therapy in the management of familial Mediterranean fever has provided new direction to investigations into the pathogenesis of this disease. Examination of HLA antigen frequencies in 53 patients with familial Mediterranean fever and appropriate controls, as well as various immunologic studies have yielded no significant differences. However, B lymphocyte typing and assays for immune complexes, lymphokines and prostaglandins may be of potential interest. Preliminary studies indicate that leukocytes of patients with familial Mediterranean fever release increased amounts of lysozyme (P<0.01), when subjected to high temperatures, and of both lysozyme and myeloperoxidase at low osmotic concentrations. The known and potential effects of colchicine on leukocyte and cellular metabolism, and the current status of colchicine prophylaxis are reviewed. In patients receiving an optimum colchicine dose of 1.5 to 1.8 mg per day, side effects have been minimal and the frequency of attacks has been decreased significantly.     

Augmented therapeutic results elicited by splenectomy in combined modality treatment of spontaneous murine breast cancer

Summary Because it had been reported that splenectomy produces a tumor-inhibitory effect in several transplantable tumor systems when the surgery is performed before tumor challenge, we attempted to examine this putative immunological manipulation in a therapeutic situation.A spontaneous, autochthonous, murine breast tumor system was utilized in the present studies, and treatment was initiated in animals bearing large tumors (averaging 0.5 g). To amplify any immunological benefit ensuing from splenectomy, the tumor burden in the host was reduced by ancillary treatment with enucleative tumor surgery or with enucleative tumor surgery plus cytoreductive combination chemotherapy.Splenectomy performed in conjunction with enucleative tumor surgery was associated with an increment of cure in each of four separate experiments in comparison to treatment with enucleative tumor surgery alone. In four of five experiments utilizing different combinations or schedules of chemotherapeutic agents following enucleative tumor surgery, the addition of splenectomy resulted in a decrease in the rate of tumor recurrence as well as an increment in the cure rate. In the fifth experiment, splenectomy resulted in a decrease in the rate of tumor recurrence, but did not effect the ultimate cure rate.Although the nature of the immunological changes resulting from splenectomy are incompletely defined at present, these results provide encouragement in the search for immunological treatments for solid tumors.This work was supported in part by Contract No. N01-CM-73703 and Grant IR01CA-14768-01A1, both from the National Cancer Institute, National Institutes of Health, Department of Health, Education and Welfare, USA, and in part by a grant from the Chemotherapy Foundation of New York, Inc.     

The natural occurrence of insulin receptors in groups on adipocyte plasma membranes as demonstrated with monomeric ferritin-insulin

This study was designed to document whether the reported distribution of insulin receptors in small groups of receptor sites randomly distributed in the glycocalyx of adipocytes and isolated adipocyte plasma membranes was a naturally occurring phenomena or due to artifacts. Possible artifacts include: (1) oligomeric forms of ferritin in the ferritin-insulin preparation, (2) an uneven distribution of the glycocalyx on the plasma membrane, or (3) ligand-induced aggregation of occupied receptor complexes. Biogel A 1.5m chromatography of the ferritin-insulin conjugate revealed the ferritin in the ferritin-insulin complex to consist of 55% monomers, 15% dimers, and 30% oligomers. The monomer peak was purified (> 95%) for use in these studies. Cationic ferritin, a glycocalyx marker, when incubated with paraformaldehyde-fixed plasma membranes, was found to be uniformly distributed on the surface of the plasma membrane indicative of uniformly distributed glycocalyx. The ability to demonstrate and inhibit ligand-induced aggregation on the isolated plasma membrane was established with a multivalent ligand, ferritin-concanavalin A. More than 66% of the ferritin-concanavalin A receptors were found in large clusters of 5 or more and 34% as singletons or clusters of up to 4 when incubated at 24°C with fresh membranes. Only 38% of the ferritin-concanavalin A receptors were in large clusters; 62% were singletons or clusters up to 4 on membranes prefixed with paraformaldehyde before incubation. The distribution of the monomeric ferritin-insulin was similar on both adipocytes and purified adipocyte plasma membranes and was consistent with earlier reports with ferritin-insulin. The quantitative distribution of the monomeric ferritin-insulin as singletons or in groups of 2–6 was comparable between the intact cells and isolated membranes incubated at 24°C. The binding of 500 μUnits monomeric ferritin-insulin per ml to the isolated plasma membranes was studied under incubation conditions similar to those used with ferritin-concanavalin A. Under all three conditions, fresh membranes at 24°C and 0–4°C and prefixed membranes at 24°C, the pattern of distribution of the monomeric ferritin-insulin as singletons or groups of 2–6 was identical, indicating that the ligand was not causing aggregation into clusters as did the concanavalin A. Thus, the occurrence of insulin receptors in small groups appears to be a natural phenomenon in the plasma membrane structure of adipocytes.     

Evidence for interactions between the energy-dependent transport of sugars and the membrane potential in the yeastRhodotorula gracilis (Rhodosporidium toruloides)

Summary A membrane potential (inside negative) across the plasma membrane of the obligatory aerobic yeastRhodotorula gracilis is indicated by the intracellular accumulation of the lipid-soluble cations tetraphenylphosphonium and triphenylmethylphosphonium. The uptake of these ions is inhibited by anaerobic conditions, by uncouplers, by addition of diffusible ions, or by increase of the leakiness of the membrane caused by the polyene antibiotic nystatin. The membrane potential is strongly pH-dependent, its value increasing with decreasing extracellular proton concentration. Addition of transportable monosaccharides causes a depolarization of the electrical potential difference, indicating that the H⁺-sugar cotransport is electrogenic. The effect on the membrane potential is enhanced by increasing the sugar concentration. The half-saturation constants of depolarization ford-xylose andd-galactose were comparable to those of the corresponding transport system for the two sugars. All agents that depressed the membrane potential inhibited monosaccharide transport; hence the membrane potential provides energy for active sugar transport in this strain of yeast.     

BRAIN pH MEASUREMENTS USING A DIFFUSIBLE, LIPID SOLUBLE pH SENSITIVE FLUORESCENT INDICATOR

Brain tissue pH was measured in cats at normocapnia hypocapnia, hypercapnia, and death from anoxia using a pH sensitive fluorescent indicator (umbelliferone) with both molecular and ionic fluorophors. A ratio analysis of the indicator's calibrated 450 nm fluorescent tissue washout curves from 340 and 370 nm excitation permitted direct determinations using a nomogram. Possible errors in these measurements related to differential quenching, absorption, and changes in the redox state of the indicator were investigated in vitro and in vivo for brain tissue and blood. In animals with preserved autoregulation, brain pH varied linearly with arterial pH (art pH 7.0, brain pH 6.98: art pH 7.4, brain pH 7.24). Brain pH at death fell to 6.68. An analysis of the indicator clearance curves suggests these measurements reflect a component of the intracellular space and the lipid solubility of the indicator suggests this is a membranous component.