Differential susceptibility of strain 2 and strain 13 guinea pigs to induction of experimental autoimmune thyroiditis. Twenty-seven of 30 Strain 2 guinea pigs immunized with 100 mug guinea pig thyroglobulin (GPTG) prepared either from Hartley (H) or Strain 13 (13) thyroids developed experimental autoimmune thyroiditis (EAT) whereas only 1 of 39 similarly immunized Strain 13 guinea pigs developed EAT. F1 hybrids behaved like the Strain 2 parent in that 16 of 20 developed EAT. The susceptible Strain 2 and F1 guinea pigs had higher hemagglutinating antibody titers to GPTG than the Strain 13 guinea pigs whereas all three strains had similar delayed skin reactivity to GPTG. The results suggest that susceptibility of guinea pigs to thyroiditis is genetically controlled and that production of antithyroglobulin antibody may be important in determining disease susceptibility.


Micro-radioimmunoassay for antibodies to Newcastle disease virus in the chicken. An indirect micro-radioimmunoassay is described in which chicken anti-Newcastle disease virus antibody was detected, with radioactively labeled rabbit anti-chicken Fab, on virus-infected microcultures of chick embryo fibroblasts. Newcastle disease virus-infected microcultures were formalin fixed and stored at 4 degrees C for up to 4 months without affecting the sensitivity of the test. The micro-technique was found to be highly sensitive and specific assay of anti-viral antibody and may allow detection of immunoglobulin class of anti-Newcastle disease virus antibody.


Serum IgE levels during the potentiated reagin response to egg albumin in rats infected with Nippostrongylus brasiliensis. Rats were immunized with 10 mug EA and alum; a low titer IgE anti-EA response was observed; subsequent infection with N. brasiliensis greatly potentiated this response. Serum from these rats was tested for its IgE content by a recently developed radioimmunoassay for rat IgE. The average serum level of IgE was 1.15 mug/ml; this level was not altered by immunization, but infection with N. brasiliensis was accompanied by an 80-fold increase in concentration of serum IgE. This increase could not be accounted for by IgE anti-EA or anti-worm antibodies. Infection with N. brasiliensis appears to have an adjuvant effect on IgE antibody responses to diverse environmental antigens as well as the experimentally induced IgE anti-EA response.


Tolerance and contact sensitivity to DNFB in mice. IV. Desensitization as a manifestation of increased proliferation of sensitized cells. Contact sensitivity to 2, 4-dinitro-1-fluorobenzene (DNFB) in mice could be rapidly and specifically suppressed by a single i.v. injection of DNBSO3 just before the elicitation skin test. The effects of desensitization were also demonstrated in vitro by the diminished response of cultured desensitized lymph node cells to specific antigenic stimulation (DNBSO3). The effect was dependent on the dose of DNBSO3 injected, and it occurred immediately and disappeared within 2 days. We believe that the mechanism is a functional defect of antigen-reactive cells which have been triggered to proliferate by the desensitizing antigen. These rapidly cycling cells may not migrate properly or respond adequately to the skin test challenge and are also refractory to additional antigen stimulation in vitro. The mechanism so postulated is derived from the finding that there was an inverse relation between the proliferative rates of the cells and their degree of specific reactivity to antigen. When the desensitizing antigen raised the proliferative rate of lymph node cells, the specific reactivity of both the whole animal and of the isolated lymphoid cells was low and vice versa.


Complement-dependent alterations in the handling of immune complexes by NZB/W mice. Serum of normal mammals contains factors which can release antigen-antibody complexes from the surfaces of leukocytes and platelets. This "complex release activity" (CRA) is mediated by the alternative pathway of complement activation, and is measured by a kinetic assay of the release of lymphocyte-bound soluble immune complexes. CRA activity was measured in the sera of NZB/W mice, which develop an autoimmune disease with aging. CRA in these mice declined rapidly after 16 weeks of  age, and by 32 weeks was barely detectable. Serum C3 levels also declined in these mice. An association test correlating CRA and C3 concentration was highly significant. The decreased complement levels in the older mice led to profound changes in their handling of injected soluble immune complexes, as demonstrated in studies of the kinetics of distribution of the complexes between plasma and circulating cells. These abnormalities may be related to the diseases which occurs spontaneously in NZB/W mice, as well as in human immune complex diseases.


Antibody-dependent lymphocyte cytotoxicity in the murine sarcoma virus system: activity of IgM and IgG with specificity for MLV determined antigen(s). Antisera with specificity for Moloney leukemia virus-(MLV) determined antigen(s) were studied for their ability to induce MLV antigen bearing target cell reduction by lymphocytes in microcytotoxicity assays. Sera from animals which had regressed Moloney sarcoma virus (MSV) tumors as well as sera from animals with progressively growing MSV tumors were found to induce normal lymphocytes to be active against the targets. Regressor serum was found also to induce cytotoxicity by immune lymphocytes from a tumor-bearing animal 15 days after MSV and from a regressor 50 days after MSV infection. Both the 19S and 7S Sephadex G-200 fractions of the antisera were found to induce cytotoxicity by normal lymphocytes and to potentiate the cytotoxicity of MSV immune lymphocytes. These activities were shown to be IgM and IgG, respectively, by the use of Sepharose-coupled anti-mouse IgM and anti-mouse IgG columns. All activity was removed by passing sera over both columns.


Serologic and biochemical studies of the Ia system of the mouse H-2 gene complex. Further evidence for I-C subregion. Three antisera were made against B10.A(5R) tissues to show the presence of an I-Cd subregion in that strain: 1) B10 anti-B10.A(5R) [anti-Dd, I-Cd]; 2) (B10 times C3H.9L) F1 anti-B10.A(5R) [anti-Ddi1; 3) (B10 times HTI)F1 anti-B10.A(5R) [anti-I-Cd]. Serologic analyses of the antisera clearly showed the presence of an Id segment in B10.A(5R) and provided further evidence for the existence of a new I-C subregion. These anti-I-Cd sera define the Ia.7 specificity and analyses of several recombinants map Ia.7 within the I-C subregion. Ia.7 is present in I-Ck, I-Cd and Ip strains. Biochemical analyses showed that the Ia.7 molecule has a molecular weight of 31,000 and is distinct from the 46,000 m.w. H2K or D molecules. The possible roles of the I-C region in immune response, mixed lymphocyte reaction, and graft-vs-host reaction are discussed.


Malaria (Plasmodium knowlesi) merozoites: immunity and the surface coat. Immune serum agglutinated merozoites of Plasmodium knowlesi in culture. Agglutinated merozoites attached to erythorcytes but were usually unable to invade. Marked aggregation of merozoites was associated with reduced invasion of erythrocytes in these cultures. The agglutination and reduced invasion were immunologically specific, for the effect of serum was greatest against homologous strains of P. knowlesi. Merozoite agglutination was caused by the binding of surface coats on adjacent parasites. This coat appeared on the plasma membrane of merozoites after it was exposed to culture medium, both with and without immune serum. The coat consisted of protein or glycoprotein, since it was susceptible to trypsin treatment. It appears that antibodies directed against this surface coat are crucial for reduced invasion of erythrocytes, at least in vitro.


The nature of murine immune response to nucleic acids. The antibody response of mice to nucleic acids appears to be genetically regulated, and may show varying patterns. SJL/J mice and AKR/Cu mice are high responders to denatured DNA complexed with MBSA and produce 7S as well as 19S antibodies. DBA/2 mice respond with low titers to this immunogen which consists of 19S antibody exclusively. C57BL/6 mice are high responders to denatured DNA, but produce only 19S antibodies. Denatured DNA-MBSA is immunogenic when injected without complete Freund's adjuvant, thereby suggesting that this antigen might be thymus independent. The pattern of immune responses of some strains to the polyribonucleotides poly(I)-poly(C) and poly(A)-poly(U) is opposite to that observed with denatured DNA. Thus, DBA/2 mice are high responders and SJL/J mice are low responders. However, the low responders SJL/J mice produce 7S as well as 19S antibody which suggests that different mechanisms operate for the regulation of immune responses to the two types of nucleic acids. Whereas no antibody is elicited upon immunization with DNA in the absence of a carrier protein, poly(A)-poly(U) and poly(I)-poly(C) are immunogenic when injected without MBSA. Immunization with complexes of the polyribonucleotides and MBSA enhanced the responses to these immunogens; however, the strain differences were not completely abolished. Poly(A)-poly(U) did not enhance the low immune responses of DBA/2 mice to denatured DNA when injected i.v. following immunization with the DNA-MBSA. However, simultaneous immunization of DBA/2 and SJL/J mice with poly(A)-poly(U) and denatured DNA-MBSA resulted in high antibody titers to both immunogens in either mouse strain.


Delayed clearance of morphine from the circulation of rabbits immunized with morphine-6-hemisuccinate bovine serum albumin. Morphine clearance from the circulation of normal rabbits and rabbits with circulating anti-morphine antibody was studied. Individual animals were injected with 14C trace-labeled morphine in saline (6 mg morphine/kg body weight). The rabbits were bled at various times after morphine injection and the amount of morphine present in the serum at each time interval was determined. Morphine could be detected in the serum of normal animals for 1 week following injection and up to 12 weeks following injection in at least one animal with antibodies to morphine. The rate of morphine clearance in animals immunized to morphine-6-hemisuccinate-bovine serum albumin was not significantly different from normal during the first 4 hr after morphine injection. However, by 24 hr after injection, the rate of morphine clearance in morphine-6-hemisuccinate-bovine serum albumin-immunized animals was significantly slower than in normal animals. The amount of morphine present in the sera of the rabbits 24 hr after injection was related to the antigen-binding capacity and independent of antibody avidity. However, the rate of morphine clearance at times greater than 24 hr after injection was related to the average antibody avidity and not related to the antigen-binding capacity.


A new immunoassay for C3. Application of the cell-linked antigen radioactive antibody (CLARA) technique. The cell-linked antigen radioactive antibody (CLARA) method of immunoassay was applied to the C3 component of the complement system. C3 from whole complement was fixed on erythrocytes sensitized with IgM antibody. The IgG1 fractions of monospecific sheep anti-C3 were labelled with 125I and measurement of their specific binding to C3-bearing erythrocytes was used to quantitate the relative amount of fixed C3. Competititive inhibition of this specific binding was used to assay fluid phase C3. 0.3-0.6mug/ml of murine C3 were detected in the supernatant fluid of in vitro cultures of mouse spleen cells.


Simple and sensitive test for the determination of phenolic compounds in urine and its application to melanoma. A simple procedure is outlined for the determination of phenolic compounds in urine.  The method involves the diazotization of p-nitroaniline and subsequent coupling of the diazonium chloride with phenolic compounds.  The diazo compound formed is determined by measuring the absorbance at 400 nm.  The extinction coefficients for several compounds have been determined.  Application of this method to determine the levels of phenolic compounds in urine in patients with melanoma showed that the assay may be of diagnostic value.  The amounts of phenols in 24-hr urine samples from patients with melanoma, patients with various other diseases, and healthy individuals were determined.  A statistical analysis of these results showed that the results for patients with melanoma were higher than those for healthy individuals and patients with other diseases at 1 percent probability level.  A comparison of this test with three other methods (Thormählen test, ferric chloride test, and ferrocitrate test) for determination of melanogens showed that the diazo test is the most sensitive one for detecting melanoma.


Absolute concentrations of dithranol and triacetyl-dithranol in the skin layers after local treatment: in vivo investigations with four different types of pharmaceutical vehicles. The pharmacokinetics of dithranol (anthralin) and its triacetate were investigated by employing a method which determines the quantity of the drug that penetrated into single layers of the human skin in vivo.  For this purpose, tritium-labeled dithranol or triacetyl-dithranol was incorporated into four different ointments.  The ointments were applied to the skin and biopsies were taken after 10, 30, 100, and 1000 min.  The horny layer was removed before biopsy by Scotch tape stripping.  The biopsies were sliced horizontally and the tritium determined in each sample.  Dithranol as well as its triacetate penetrated best from more hydropholic ointments (Vaseline and aqueous wool-wax-alcohol ointment).  From hydrophilic ointments (polyethylene glycol ointment and aqueous hydrophilic cream), only poor penetration was observed.  Dithranol penetrated in far greater amounts than its triacetate, and the two compounds revealed fundamentally different penetration kinetics in epidermis and dermis.  The data indicate that the triacetate was not split into its parent compound, dithranol, in substantial quantity, as the data obtained show the criteria of two independent substances.


DNA repair in tumor cells from the variant form of xeroderma pigmentosum. Cells from most patients with xeroderma pigmentosum (XP) can be shown to be defective in repairing ultraviolet (UV) light-induced damage to their DNA, for they have a reduced rate of UV-induced thymidine incorporation.  XP variants, however, have clinical manifestations of XP, but all their tissues tested to date have a normal rate of UV-induced 3H-thymidine incorporation. We have now tested tumor cells from an XP variant and from a typical XP patient.  The variant's tumor cells, in contrast to those of the typical patient, had no detectable defect in their UV-induced thymidine corporation.  We conclude, therefore, that the cells that formed tumors in this XP variant resemble his other cells in DNA repair capacity, and do not represent a minor cell population with the kind of DNA repair defect that is reflected in reduced UV-induced thymidine incorporation.


IgE in human eccrine sweat.

IgE in human eccrine sweat. By the use of a sensitive and specific double antibody radioimmunoassay, immunoglobulin  E was demonstrated in sweat from 6 of 11 healthy volunteers; the concentrations of IgE ranged from 1 to 3.9 ng/ml of sweat.  Six of seven patients with dermatitis and elevated levels of serum IgE also had high IgE levels in sweat; the highest IgE value in sweat was 75.5 ng/ml and was noted in a patient with atopic dermatitis. Thus, all the main immunoglobulin classes can be demonstrated in eccrine sweat.


Effect of polysorbate 85 on human skin.

Effect of polysorbate 85 on human skin. Ten percent polysorbate 85, a nonionic surfactant, was applied on the upper arm of 15 healthy individuals under occlusive dressing daily for 4 days.  The other arm was similarly treated with the ointment base (white petrolatum USP) to provide the control area.  At the end of treatment, macroscopic observations indicated minor erythema in 11 cases and no visible changes were noted on the surfactant-treated areas of 4 persons or any of the control areas.  No definite histologic changes were observed by microscopic evaluations.  The results of biochemical assays, however, were more definitive. The content of the epidermal phospholipids was elevated within a range of 5 to 65 percent as a result of the treatment with polysorbate 85 preparation.  Radioactive tracer studies indicated higher rates of 32P incorporation into epidermal phospholipids, TCA-soluble, DNA and RNA fractions of the surfactant-treated skin.  Results resemble those that were documented in earlier studies with rabbit skin.


The innervation of hyperplastic epidermis in the mouse: a light microscopic study. The innervation of the skin of hairless mice has been studied following induction of epidermal hyperplasia by physical and chemical methods. Physical stimuli comprised ultraviolet irradiation, heat, wounding, and friction. Effective chemicals included benzene, carbon tetrachloride, chloroform, creosote, formaldehyde, hexadecane, hydrobromic acid, sodium lauryl sulfate, and turpentine. Epidermal hyperplasia, however produced, was associated with growth of sensory nerve fibers into the outer part of the epidermis. Following a single 10-min exposure to an ultraviolet sunalmp at 40 cm, the nerves extended into the epidermis within 24 hr and disappeared during the second week as the epidermis reverted to its normal thickness. Repeated irradiation (until tumors appeared) was accompanied by persistent hyperplasia and neural invasion. Of 32 papillomas examined, intraepithelial nerves were found in 28. The presence and location of nerves in the tumor epithelium were related to the incorporation of tactile hair disc epithelium. The hyperplastic regenerative epithelium at the margins of skin ulcers were also invaded by nerves which sometimes followed the migrating epithelium across the ulcer floor. Since the regenerative epithelium was not directly treated, it was concluded that the proliferation of nervous tissue in response to skin injury was the result of the hyperplasia per se, regardless of the method used to produce it.


The detection of carotenoid pigments in human skin. Carotenoid pigments were extracted chemically from both epidermis and dermis obtained from non-carotenemic individuals at autopsy.  Absorption maxima characteristic of beta-carotene were found in the extracts of specimens of epidermis following cantharidin application in volunteers made carotenemic by the ingestion of beta-carotene (180 mg/day for 10 weeks).  These maxima were absent in the extracts of epidermis obtained from the volunteers before beta-carotene ingestion.


Effect of heparin in experimental gram-negative septicemia. Young rabbits were given live encapsulated or heat-hilled Pasteurella multocida by the intraperitoneal route. Those animals given live organisms regularly developed clinical illness, positive blood cultures, positive limulus lysate tests on plasma, leukopenia, thrombocytopenia, and disseminated intravascular coagulation and died. Death occurred in 68% of these rabbits by 12 hr after infection and in 100% by 24 hr. Septicemic animals given heparin as an anticoagulant similarly developed clinical illness, leukopenia, and thrombocytopenia, but no laboratory evidence of disseminated intravascular coagulation. The mortality rate in these animals was  90% 12 hr after infection and 100% 24 hr after infection. None of the animals given heat-killed organisms developed leukopenia, thormbocytopenia, or disseminated intravascular coagulation, and none died. The data show that the survivial rate in septicemic rabbits was not improved with heparin, although fibrinogen consumption (disseminated intravascular coagulation) was abolished.


Hepatitis A: report of a common-source outbreak with recovery of a possible etiologic agent. I. Epidemiologic studies. From the end of October to the end of December 1973, an epidemic of hepatitis A affected 40 students at a state university, 11 employees of two restaurants, and 11 other residents in a southwestern metropolitan community. Two distinct epidemic waves of illness occurred, and investigation implicated two local restaurants as sources of infection, one for each epidemic wave. An index patient who prepared food was identified at each restaurant, and the two restaurants were linked by the fact that the index patient who worked at the second restaurant had patronized the first restaurant. Foods implicated as vehicles of transmission included 'guacamole,' green salad, spaghetti, and hamburger. The findings represent a rare example of a 'double' common-source outbreak with two distinct epidemic waves related to two separate eating establishments.


Hepatitis A: report of a common-source outbreak with recovery of a possible etiologic agent. II. Laboratory studies. During investigation of a food-borne outbreak of hepatitis A among university students in a southwestern metropolitan community, immune electron microscopic examination of a concentrated stool suspension pooled from seven acutely ill individuals revealed viruslike particles 17-nm in diameter. These particles were initially coated by antibody contained in the convalescent-phase serum of one of the ill students as well ad by antibody in convalescent plasma of a prison volunteer originally infected with the MS-1 strain of hepatitis A virus. Rises in titer of antibody to this particle were demonstrated by immune electron microscopy in acute and convalescent sera from student patients as well as in pre-inoculation and convalescent sera from the prison volunteer. Two chimpanzees inoculated intravenously with the concentrated preparation of pooled human stools developed viral hepatitis. During acute illness their feces contained particles morphologically identical to those in the inoculum. These findings represent the first reported recovery of the presumed etiologic agent of hepatitis A from a naturally occurring community outbreak of disease in the United States.


Effects of cyclophosphamide on hepatitis B virus infection and challenge in chimpanzees. The role of antibodies to the surface antigen of hepatitis B virus (HBsAg) and to the hepatitis B core antigen (HBcAg) and of cell-mediated immunity in hepatitis B virus infections and in resistance to challenge was studied in six chimpanzees. Two chimpanzees were studied during primary hepatitis B virus infection while being treated with cyclophosphamide (5-15 mg/kg im three times per week). Cyclophosphamide treatment may have predisposed these chimpanzees to chronic hepatitis B virus infections. Four other chimpanzees convalescent from primary hepatitis B virus infection (subtype ayw) were challanged with hepatitis Bvirus; two were callenged with heterologous virus (subtype adw) without immunosuppression, and two were challenged with homologous virus (subtype syw)while being treated with cyclophosphamide. Chimpanzees with residual humoral immunity, demonstrable by serum antibodies against the group determinant a and subdeterminant y of HBsAg, resisted challenge with either heterologous or homologous hepatitis B virus despite cyclophosphamide immunosuppression in the latter case. Antibodies to the hepatitis B virus core did not appear necessary for protection against hepatitis B virus infection in these chimps.


Isolation and purification of hepatitis B antigen by electrochromatography. The electrochromatography column is a unique glass device designed to function both as a chromatography column and as an electrophoresis column; it can also be operated so that both column chromatography and electrophoresis can take place in one simultaneous operation (electrochromatography). When the column is used for electrochromatography, proteins of the same molecular size, but with different electrophoretic mobilities, will be separated by electrophoresis, whereas proteins with the same electrophoretic mobilities but of different molecular size will be separated by molecular sieving. If proteins vary only slightly in these physical properties, separation is greatly enhanced by the action of the electric field on the filtering molecules. Thus, electrochromatography allows proteins to be separated simply, rapidly, and with greater resolution than other systems currently available. Electrochromatography has been used successfully for the purification of hepatitis B antigen, which is not separated from closely related serum immunoglobulins by either chromatography or electrophoresis. This method may become an important tool for the isolation and purification of a large number of proteins and viruses not easily separated by means of current technology.


Antimicrobial therapy of experimental endocarditis caused by Staphylococcus aureus. The rate at which various antimicrobial agents eradicated Staphylococcus aureus from cardiac vegetations in a rabbit model of endocarditis was studied. The rate at which various drugs and combinations killed high titers of bacteria in broth correlated with the relative effectiveness of the agents in vivo. Gentamicin plus penicillin proved to be synergistic in vitro and more effective in eradicating bacteria from cardiac vegetations in vivo than was penicillin alone. Vancomycin killed bacteria at a rate similar to that for the combination of penicillin and gentamicin, and the rate for cefazolin was similar to that for penicillin alone. Clindamycin was less effective in vivo and in vitro than penicillin. Therapy with rifampin led to the emergence of resistant organisms, and, when penicillin, this drug was less effective in vitro and in vivo than was penicillin alone. This model appears to offer an effective method for evaluation of antimicrobial treatment of staphylococcal endocarditis.


Teratogenic effects of Colorado tick fever virus in mice. The teratogenic effects in mice of Colorado tick fever (CTF) virus, a human pathogen, were demonstrated. Various routes of inoculation, gestational periods, and dosages of virus were studied. The incidence of stillbirth and neonatal death increased significantly after inoculation of pregnant mice with CTF virus. CTF virus crossed the placenta and replicated to high titers in the placental and fetal tissues. Mice that survived the first six weeks of life had neutralizing antibody to CTF virus. Neutralization tests, in which pregnant mice were inoculated with both CTF virus and specific antiserum to CTF virus, validated the teratogenic effects of CTF virus in mice.


In vitro correlates of cell-mediated immunity in human tonsils after natural or induced Rubella virus infection. The techniques of hemagglutination inhibition and in vitro lymphocyte transformation and the assay of migration inhibitory factor were used for study of the development of rubella antibody activity in serum and tonsillar tissue washings and of the appearance of rubella-specific, cell-mediated immunity in circulating and tonsillar lymphocytes in groups of children who underwent tonsillectomy at various intervals after natural rubella infection,  subcutaneous immunization with HPV-77 DE/5 vaccine, or intranasal inoculation with RA27/3 live, attenuated rubella virus vaccine. Antibody response in serum and tonsillar tissue washings was detected regularly after natural infection or immunization. Development of specific cell-mediated immunity in circulating lymphocytes was regularly observed after natural infection and frequently after immunization. Natural infection or intranasal immunization with rubella vaccine resulted in the appearance of cell-mediated immunity in the tonsillar lymphoid tissue, and the response was detectable up to several years after natural infection in several cases. It is significant, however, that the level of cell-mediated immunity in tonsils was conspicuously low after subcutaneous immunization. These data suggest the induction of local cell-mediated immunity in tonsillar lymphoid tissue after local mucosal application of rubella virus.


The generation of superoxide anion by various types of phagocyte. Polymorphonuclear leukocytes derived from human peripheral blood, rabbit peritoneal exudates, and guinea pig peritoneal exudates generate measurable quantities of superoxide anion when challenged with opsonized zymosan, but not when challenged with zymosan that has not been opsonized. The generation of superoxide is dependent upon the concentration of cells and closely parallels the stimulation of the hexose monophosphate shunt in the phagocytes. In contrast, no superoxide can be detected when rabbit alveolar macrophages (either normal or activated by prior vaccination of the animal with bacille Calmette-Guérin) or rabbit peritoneal macrophages are similarly treated. These data suggest that although superoxide anion may be involved in the bactericidal activity of the polymorphonuclear leukocyte, it is less likely to play a significant role in the bactericidal activity of the macrophage.


Generalized Yersinia enterocolitica infection. The clinical course of generalized Yersinia enterocolitica infection in 13 patients was observed. Two syndromes were apparent; seven patients had an acute septicemic picture, whereas the rest had a subacute localizing infection that usually produced hepatic or splenic abscesses. The latter form often resembled amebic hepatitis clinically, and five of the six patients died despite antibiotic treatment. The acute septicemic form of the disease resembled systemic salmonellosis, and most cases responded to therapy with the appropriate antibiotics.


Short-term metabolism of (14-C) tryptophan in rats infected with Trypanosoma brucei gambiense. The pathways of short-term tryptophan metabolism in rats infected with Trypanosoma brucei gambiense were studied to determine whether this amino acid undergoes rapid conversion to tryptophol and indole lactic acid, two metabolites produced in vitro by T. brucei gambiense. Intravenously administered L-(3-14-C)-tryptophan was metabolized to at least two compounds in infected rats: tryptophol (indole ethanol) and indole acetic acid. Indole lactic acid was not detected as a metabolite. Approximately 76 percent of the labeled substrate was metabolized along these pathways in 30 min. Control rats exhibited no detectable metabolism of tryptophan during the same period. These results indicate that dietary tryptophan may be diverted from the kynurenine and serotonin pathways by parasite metabolism to compounds possessing potential pharmacological or toxic activity.


Kinetics of hemodiafiltration. I. In vitro transport characteristics of a hollow-fiber blood ultrafilter. The transport characteristics of a hollow-fiber blood ultrafilter were studied in vitro to provide an understanding of the factors which control solute removal rate and to permit design of a full-scale clinical device. The dependence of ultrafiltrate flux on transmembrane pressure difference, protein concentration, flow rate, and fiber geometry was correlated in terms of available theoretical analyses. Solute rejection was 1.0 for albumin and decreased to nearly zero for solutes of several thousand molecular weight. An analysis of overall hemodiafilter behavior showed that blood and ultrafiltrate flow rates of 200 ml. per minute can be attained with a device of reasonable size which would provide an inulin whole blood clearance of about 100 ml. per minute.


The effect of inosine, pyruvate, and inorganic phosphate on 2,3-diphosphoglycerate, adenine, and hypoxanthine nucleotide synthesis in outdated human erythrocytes. Phosphate compounds of red cells stored at 4 degrees C. in acid-citrate-dextrose solution for 15 to 44 days, incubated at 37 degrees C. for 2 and 4 hours with inosine, pyruvate, and inorganic phosphate, and then stored for 2 weeks have been examined by ion-exchange column chromatography. After 2 and 4 hours of incubation the 2,3-diphospholycerate level increased to 1,385 and 1,819 mumoles per 100 ml. of erythroeytes, respectively. After 4 hours of incubation the content of AIP increased to about 80 per cent of the physiologic level, while the content of AMP and ADP decreased considerably. These data suggest that the elevation of ATP is in part connected with the phosphorylation of AMP and ADP. The incorporation of inosine or hypoxanthine into the IMP and ITP has been proved. Four hours incubation period caused the synthesis of IMP and ITP to the mean value of 187 and 141 mumoles per 100 ml. of erythrocytes, respectively. No evidence of IDP formation was observed during the chromatographic separations of erythrocytic phosphate compounds. Storage of the erythrocyte suspension which has been incubated for 4 hours leads to the slower decomposition of 2,3-diphosphoglycerate, ATP, ITP, and IMP, and to the increase of AMP and ADP.


Enhancement of filterability of sickle cells by cyanate: an effect independent of oxygen saturation. Filterability and morphology of cyanate-treated sickle cells were compared to those of untreated cells at equal oxygen saturations to determine whether carbamylation inhibited sickling by an effect other than by its alteration of the oxygen dissociation curve. Morphology of treated and untreated cells was not significantly different at all levels of oxygen saturation examined. Filterability, on the other hand, was improved significantly by carbamylation. This latter finding indicates that carbamylation enhances deformability of sickle cells by a mechanism(s) in addition to its effect on red cell oxygen saturation. This mechanism(s) may account for the clinical benefit of cyanate therapy with doses which do not significantly affect the oxygen dissociation curve.


Impaired renal tubular reabsorption of sodium in hypothyroid man. The renal response to acute salt loading and to stimuli for rapid sodium conservation was studied in 14 patients with untreated myxedema and in 13 euthyroid control subjects in balance on a 155 mEq. sodium intake. The salt-loading studies reveal urinary excretion of sodium in the myxedema patients within the range of controls despite reductions of 34 per cent in glomerular filtration (p less than 0.001) and 37 per cent in filtered load of sodium (p less than 0.001) in the former group. The capacity to conserve sodium in response to stimuli for rapid sodium conservation [postural change and administration of a supramaximal dose of 9alpha-fluorohydrocortisone (9alpha-F)] was impaired in patients with myxedema. The per cent decrease in sodium excretion during the upright posture in the hypothyroid patients was 28 per cent, less than half that observed in the control subjects, 62 per cent (p less than 0.005). Following administration of 2 mg. of 9 alpha-F the per cent decrease in sodium excretion was less (p less than 0.05) in the hypothyroid patients (50 per cent) than in control subjects (72 per cent). In all studies, baseline sodium excretion was comparable in both groups. Fractional excretion of sodium was significantly increased in the hypothyroid patients prior to (p less than 0.005) and during saline loading (p less than 0.05) and at the time of the subnormal responses to stimuli for acute sodium conservation (p less than 0.05 less than 0.005). Potassium excretion was reduced in the hypothyroid patients, even after 9alpha-F. These observations indicate decreased tubular reabsorption of sodium in myxedema under the experimental conditions described. The findings are most consistent with a role for thyroid hormone in normal sodium reabsorption. That this is not related to mineralocorticoid deficiency is suggested by the impaired sodium reabsorptive response to 9alpha-F.


A simplified and rapid quantitative assay for propionic and methylmalonic acids in urine. Propionic and methylmalonic aciduria occur individually in inborn errors of metabolism and together in vitamin B12-deficient states. A method is described for the simultaneous and rapid extraction of these acids from urine and their quantification by a simple gas chromatographic technique. The assay is based upon the spontaneous and quantitative decarboxylation of methylmalonic acid (MMA) at 225 degrees C. to its monocarboxylic acid product, propionic acid. By utilizing another substituted malonic acid, ethylmalonic acid, as a specific internal standard, accurate quantitation is possible by peak height ratio analysis, Endogenous propionic acid is then measured at 130 degrees C., a temperature at which methylmalonic acid does not decarboxylate. The assay is rapid with a total running time of approximately 2 hours. The method provides excellent resolution of propionic acid excretion at or above 0.5 mg. per liter of urine. The level of resolution for methylmalonic acid, in the presence of propionic aciduria, was 5 mg. per liter of urine.


Non chromatographic colorimetric assay for total taurine-conjugated bile acids: application of measurements of glycine: taurine ratio in bile. A direct colorimetric assay for total conjugated taurine from bile-rich duodenal aspirates is described. The method is based on complete acetylation of the free hydroxyl groups by acetic anhydride at 100 degrees C. of both the tri- and di-hydroxy bile acids in Folch extracted bile samples. Taurine-conjugated bile acids are measured by ion pair formation with Azure A and subsequent extraction of the complex into the organic phase of a biphasic system. Absorption at 645 nm. of this complex directly quantifies total taurine-conjugated bile acid. Total bile acids are then estimated by the 3alpha-hydroxysteroid dehydrogenase assay. The difference between the concentration of the total conjugated bile acid and of the total conjugated taurine determines the concentration of glycine conjugates and the glycine:taurine ratio. Potentially interfering materials such as sulphalipids, certain phospholipids, and unconjugated bile acids are removed by Folch extraction. The 3-hydroxysteroid sulfates (cholesteryl sulfate, lithocholate sulfate, and glycocholate sulfate) are not measurable by heating in acetic anhydride and do not interfere. Taurolithocholate-3-sulfate, under identical conditions, gives a measurable but very low color yield and in normal physiologic concentrations would contribute negligible color. As previously reported, this assay under prescribed conditions is selective for long-chain amphipathic sulfates or sulfonates with no measurable color yield with glycine conjugates, unconjugated bile acids, free fatty acids, or lecithin. Values for glycine:taurine ratios by the above-described method in both normal bile extracts and extracts from patients with either elevated or depressed ratios relate closely to those determined by thin-layer chromatography.


Sequential blood samples from the tail vein of rats and mice obtained with modified Liebig condenser jackets and vacuum. A simple method for obtaining multiple, sizeable blood samples rapidly and sequentially from mice and rats without the use of anesthesia utilizing an adapted Liebig condenser jacket connected to a vacuum line is described. The method permits the removal and insertion of sample tubes during the procedure so that anticoagulated and clotted specimens can be obtained during a single bleeding.


Effects of testosterone and prolactin or growth hormone on the accessory sex organs of castrated mice. In 5-day experiments, neither bovine prolactin (300 or 100 i.u./kg) nor ovine growth hormone (25 i.u./kg) alone significantly enhanced accessory sex organ weights in the castrated mouse. Seminal vesicle weights, and to a lesser extent anterior prostate gland weights, were augmented by the simultaneous injection of testosterone (1-5 mg/kg) daily plus prolactin or growth hormone. The effect was greater than that produced by testosterone alone. The levels of fructose in accessory sex organs used to indicate androgenic activity were similar in castrated mice receiving testosterone alone or in combination with prolactin or growth hormone. Prolactin alone did not influence uptake of (3H)testosterone by the seminal vesicles or anterior prostate gland over a 5 min period in vivo.


Oestrogen, progesterone and corticosteroid concentrations in peripheral plasma of sows during pregnancy, parturition, lactation and after weaning. Steroid hormone concentrations in plasma have been measured in blood samples taken from conscious sows with ear vein catheters. In late pregnancy, the plasma progesterone concentration ranged from 6 to 12 ng/ml and it decreased in all animals before the onset of parturition. Total unconjugated oestrogens increased to high values of up to about 3 ng/ml in late pregnancy and then declined after the onset of parturition. Oestrone was the predominant unconjugated oestrogen measured. Plasma corticosteroid (mainly cortisol) concentration was about 33 ng/ml and showed no consistent change at the time of parturition. During lactational anoestrum the plasma concentration of progesterone and total unconjugated oestrogens was very low, while that of corticosteroids was 21 ng/ml. When the piglets were weaned at 26-31 days, sows came into oestrus 4-12 days later, and this was preceded, or accompanied by, an increase in plasma oestrogens. In the luteal phase, plasma progesterone concentrations rose to 20-35 ng/ml. A sow whose piglets were removed at birth, showed signs of oestrus (vulval enlargement and a lordosis response), but a lack of receptivity to the boar associated with no detectable changes in the plasma oestrogen concentration; however, ovulation probably occurred since plasma progesterone values increased in a manner comparable to that found after the formation of normal corpora lutea in other sows. After a second non-receptive cycle, the sow was mated and became pregnant at the third post-weaning oestrus. At parturition the concentration of progesterone and total unconjugated oestrogens was greater in placental venous plasma than in maternal jugular plasma, which indicates placental synthesis of these hormones. A greater concentration of plasma corticosteroids in foetal blood than in placental venous or maternal jugular plasma suggests foetal synthesis in late pregnancy.


Superfusion in vitro in the study of ovarian steroidogenesis. A method for the continuous superfusion of porcine corpus luteum tissue is described which readily allows both the introduction of regulatory factors to the incubating tissue, and sampling of the tissue. Oestrogen (principally oestradiol) and progestin (principally progesterone) can be measured for up to 24 h in the superfusate from corpora lutea of all ages, and the secretion of both steroids is stimulated by the addition of luteinizing hormone. The pattern of response of both steroids to a pulse of gonadotrophin was similar in that a rapid transient increase in secretion occurred followed some time later by a secondary and more prolonged response. A second pulse of gonadotrophin introduced 6 h after the first also stimulated steroid secretion, indicating that during superfusion in vitro the porcine corpus luteum does not become refractory to the steroidogenic effect of gonadotrophin.


Restoration by oestradiol benzoate of a neural and hormonal rhythm in the ovariectomized rat. The rate of [35S]methionine incorporation into protein in discrete cerebral areas was measured before and after the administration of oestradiol benzoate (OB) to chronically ovariectomized rats. The circadian rhythm of incorporation which is normally seen in the intact cyclic female rat was deleted by ovariectomy. A daily rhythm of incorporation reappeared, however, in all the brain areas studied 30 h after a single injection of OB (20 mug), and was still present 12 days later. The release of luteinizing hormone (LH) after administration of 20 mug OB was measured in chronically ovariectomized animals and was found to be biphasic. High levels of LH after ovariectomy were initially reduced by negative feedback, but this phase was followed 52 h later by a facilitation of LH release between 15.00 and 18.00 h. The facilitation of LH release at this time of day was still detectable 12 days after the initial injection. The evidence for a functional link between the rhythm of neural activity which is reflected by [35S]methionine incorporation, and the ability to 'time' the facilitation of LH release is discussed.


Episodic release of luteinizing hormone in the domestic fowl. Plasma luteinizing hormone (LH) levels were measured by radioimmunoassay in serial samples taken from intact adult cockerels, gonadectomized fowl of both sexes and laying hens. By sampling at 30 and 10 min intervals, it was shown that in cockerels LH is released episodically. Each secretory episode lasted 90-120 min and was characterized by a 100-200% rise in LH levels over a period of 10-15 min followed by a more gradual, exponential decline. Mean plasma LH levels were found to be depressed in cockerels as a result of the handling associated with taking blood samples; in cases where LH levels were most depressed, an episodic pattern of secretion could not be demonstrated. In the gonadectomized fowl, an episodic pattern of LH secretion was shown when blood samples were taken at 10 or 5 min intervals. During each secretory episode, which lasted 20-45 min, LH levels rose by 20-60%. It is suggested that the increase in mean plasma LH levels in gonadectomized fowl (30-77 ng/ml) over those in cockerels (7-16 ng/ml) is related to an increase in the frequency of the episodic release of the hormone. Episodic discharges of LH could not be demonstrated in laying hens. In these birds the mean LH levels are low (1-5--2-4 ng/ml) and consequently may be maintained by secretory episodes which are of too low an amplitude to be detectable.


Plasma levels of luteinizing hormone in gonadectomized Japanese quail exposed to short or to long daylengths. Plasma levels of luteinizing hormone (LH) were measured by radioimmunoassay in gonadectomized male and female Japanese quail, exposed either to 8 h light: 16 h darkness per day (8L:16D; short days) or to 20L:4D (long days). In both sexes, exposure to long days increased LH levels and in the gonadectomized quail LH continued to rise over several weeks. Eventually the castrated quail had levels about five times higher than the control birds and the ovariectomized quail had levels about 14 times higher than their controls. Quail kept on short days had low LH levels while birds kept on long days and returned to short days resumed low levels after a delay of some days. Since very high levels of LH occurred in gonadectomized quail only when they were on long days, we conclude that the photoperiodic regulation of LH secretion does not operate solely by adjusting sensitivity to gonadal feedback, but works in a more direct manner. Ovariectomized females whether on long days (high LH) or short days (low LH) grew masculine plumage and castrated males retained male plumage. This confirms that the ovary is responsible for sexual dimorphism of plumage and shows that the action of the ovary is not mediated by LH (through feedback). The remaining rudimentary (right) gonad in ovariectomized females did not undergo visible hypertrophy and did not secrete enough hormone to stimulate the cloacal gland or oviduct.


Clearance and metabolism of neurophysins by rat kidney. The half-life in the circulation of 125I-labelled porcine neurophysins, injected intravenously into rats, was determined. The radioactivity in blood collected at intervals after injection was characterized as neurophysin by polyacrylamide gel electrophoresis. The half-life of neurophysin I was 1-5 plus or minus 0-1 (S.E.M.) min and that of neurophusin II was 1-7 plus or minus 0-1 min. The metabolic clearance rate of neurophysin I and neurophysin II was 1-9 plus or minus 0-1 and 1-3 plus or minus 0-2 ml/min/200 g respectively. Clearance from the circulation was due to both equilibration with the extravescular space and rapid specific accumulation by the kidney. The neurophysins were filtered at the glomerulus. Most of the neurophysin takken up by the kidney was incorporated into lysosomes, probably in the proximal tubule and degraded to low-molecular-weight metabolites which appeared in urine and which represented the principle mode of excretion of neurophysins.


Distribution of radioactivity in the organs of the rat and mouse after injection of (125I)alpha-melanocyte-stimulating hormone. The distribution of radioactivity after intrajugular injection of 125I-labelled alpha-melanocyte-stimulating hormone (alpha-MSH) was studied by whole-body autoradiography of the mouse and by direct measurement of radioactivity in individual organs of the rat. Very high uptake of radioactivity in the pineal gland was measured 5 min after the injection of (125I)alpha-MSH. Lower levels of accumulation of radioactivity were found in the kidney and in the posterior (including intermediate) lobe of the pituitary. High uptake was also found in the thyroid, stomach, and oesophagus. The specificity of uptake of (125I)alpha-MSH into the pineal and pituitary is suggested by the very low uptake of Na125I into those tissues.


Distribution of radioactivity in the organs of the rat and mouse after injection of L-(3H)prolyl-L-leucyl-glycinamide. The distribution of radioactivity after intrajugular injection of L-(3H)-prolyl-L-leucyl-glycinamide has been studied by whole-body autoradiography in the mouse and by direct measurement of radioactivity in individual organs of the rat. There is good agreement between results obtained with the two techniques and animal species. High levels of radioactivity were found in the pineal gland, anterior pituitary, posterior (including intermediate) lobe of the pituitary, and epididymal and brown fat. Lower uptake of radioactivity occurred in the submaxillary gland, kidney, and adrenal gland. The preferential uptake of radioactivity by the pineal gland after injection of the labelled tripeptide suggests a role for this hypothalamic hormone in the control of pineal activity.


Pituitary hormone secretion in the genectically male rat pseudohermaphrodite. Pituitary content or concentration of follicle-stimulating hormone (FSH), prolactin and growth hormone in the genetically androgen insensitive male rat pseudohermaphrodite is intermediate between normal male and female rats, while pituitary luteinizing hormone (LH) concentration and serum FSH levels are the same as in the normal male. The concentration of serum LH, prolactin and growth hormone indicated no sexual dimorphism. Although the pseudohermaphrodite is genetically male with a female phenotype, our results suggest some degree of masculinization of the hypothalamic-pituitary system.


A comparison of the characteristics of parturition induced by prostaglandin F2alpha, infused intra-aortically, with those following ovariectomy in the rat. Prostaglandin F2alpha (PGF2alpha) was infused intra-aortically at different rates into rats for 11-5 h from day 18 of gestation. At doses in excess of 5 mug/h premature delivery invariably occ'rred 40-41 h later. At lower doses an all-or-none effect was observed, some animals delivering prematurely and others continuing to term. Parturition could not be induced earlier than 40 h after the start of the treatment even by a continuous infusion of PGF2alpha, suggesting that the luteolytic role of PGF2alpha is more important than its oxytocic action in this case. The rate of fall in plasma progesterone was greater in the rats delivering prematurely than those delivering at term, although the final concentrations of between 15 and 30 ng/ml at delivery were the same. Labour appeared more normal in the animals receiving PGF2alpha than in those which had been bilaterally ovariectomized. The possible significance of this in relation to an effect of PGF2alpha on oestrogen secretion is discussed.


Partial feminization of hepatic steroid metabolism in male rats after neonatal administration of cyproterone acetate. The metabolism of (4-14C)4-androstene-3,17-dione, (4-14C)5alpha-androstane-3alpha, 17beta-diol and (1,2-3H)5alpha-androstane-3alpha, 17beta-diol 3,17-disulphate was studied using the microsomal fraction and the metabolism of (4-14C)4-androstene-3,17-dione was studied using the 105 000 g supernatant fraction of liver from male and female rats aged 5 months that had been treated with cyproterone acetate before (from day 13 of pregnancy) and after birth (until 3 weeks of age). Nearly all sex-dependent enzyme activities in the treated male rats were changes in a direction characteristic of female rats: 5alpha-reductase active on 4-androstene-3,17-dione increased in activity whereas 3beta- and 17alpha-hydroxysteroid reductases and 6beta- and 16alpha-hydroxylases active on 4-androstene-3,17-dione and 2alpha-, 2beta- and 18-hydroxylases active on 5alpha-androstane-3alpha,17beta-diol decreased in activity. Enzyme activities not under gonadal control, i.e. 3alpha- and 17beta-hydroxysteroid reductases active on 4-androstene-3,17-dione and 7alpha-hydroxylase active on both 4-androstene-3,17-dione and 5alpha-androstane-3alpha, 17beta-diol, were not affected by cyproterone acetate. The liver enzyme activities in treated female rate were generally not affected although significant effects were noted in two cases; in one of these (17alpha-hydroxysteroid reductase) a testosterone-like effect was observed. The results obtained are probably best explained in the following way: treatment with theanti-androgen during the neonatal period results in less efficient imprinting of the hypothalamo-hypophysial system leading to less pronounced masculine setting of sex-dependent enzyme levels and also to a relative androgen unresponsiveness. It is suggested that the biochemical methods used in the degree of neonatal sexual differentiation of the hypothalamo-hypophysial system than biological and psychological methods previously available.


Estimation of aldosterone, 11-deoxycorticosterone, 18-hydroxy-11-deoxy-corticosterone, corticosterone, cortisol and 11-deoxycortisol in human plasma by gas-liquid chromatography with electron capture detection. A method for determining the plasma concentrations of six major corticosteroids, aldosterone, 18-hydroxy-11-deoxycorticosterone (18-OH-DOC), corticosterone, deoxycorticosterone (DOC), cortisol and 11-deoxycortisol using gas-liquid chromatography with electron capture detection is described. Esterification of suitable derivatives of these compounds with heptafluorobutyric anhydride (HFB) allowed detection of quantitities of steroid, ranging from 0-3 pg for androstenetrione HFB (from cortisol) to 2-3 pg for corticosterone HFB. No detectable reagent blank was obtained for any compound when water was used instead of plasma and this was also the case when plasma from an adrenalectomized subject was analysed, with the exception of 18-OH-DOC where a reproducible but negligibly small blank occurred. Coefficients of variation for replicate determinations ranged from 8% for corticosterone to 17% for aldosterone. Concentrations in a series of normal human plasma samples were as follows: aldosterone, 4-0- 18-0 ng/100 ml; 18-OH-DOC, 20-16- ng/ml; corticosterone, 0-08 - 0.-80 mug/100 ml; DOC, 2-8 - 16-0 ng/100 ml; cortisol, 2-5 - 10-0 mug/100 ml; and 11-deoxycortisol, 40-0 - 400-0 ng/100 ml. When seven normal subjects were treated with dexamethasone concentrations of DOC, cortisol and 11-deoxycortisol fell to below the limit of the normal range, those of 18-OH-DOC and corticosterone were at the lower end of the normal range while the concentration of aldosterone was not significantly affected.


Effect of calcium deprivation on parathyroid hormone-mediated bone and kidney contributions to the maintenance of plasma calcium in rats. This study was designed to investigate the roles of bone and kidney in the acute regulation of plasma calcium by parathyroid hormone (PTH) during prolonged calcium deprivation. The effect of PTH was assessed by gland ablation. Animals were thyroparathyroidectomized or sham-operated and their urine was collected for 3 h. Subsequently they were anaesthetized and bled from the abdominal aorta. In rats fed on a low calcium diet, urinary hydroxyproline excretion was enhanced and, unlike animals fed on a normal diet, decreased 3 h after thyroparathyroidectomy (TPTX). In addition TPTX decreased plasma calcium by 0-45 mg/100 ml in normal rats compared with 1-94 mg/100 ml in animals fed on a calcium-deficient diet. Urinary calcium increased by 161 and 12 mug and accounted for 82 and 1-4 % of the fall in plasma calcium in normal and calcium-deprived animals respectively. The corresponding contributions of bone were 18 and 98-6%. These findings support the view that with prolonged calcium deprivation in adult rats, the relative contributions of bone and kidney to the acute regulation of the plasma calcium level by PTH are reversed. As a result, bone rather than kidney becomes the more important organ. At the same time non-PTH-mediated kidney reabsorption of calcium is increased.


Concentrations of prolactin in serial blood samples from goats before, during and after milking throughout lactation. Two experiments are described in which prolactin was measured in (a) serial blood samples taken at monthly intervals (April-November) from six lactating goats before, during and after milking and (b) in similar blood samples taken on days 2, 7 and 12 after parturition. The quantity of prolactin released at milking (April-August) varied both between goats and within the individual goat. No correlation was found between the duration of milking and the quantity of prolactin released into the circulation (r equals minus 0-24, P greater than 0-1) or the average concentration of prolactin found at milking (April-August) and the average daily milk yield of the individual goat (r equals minus 0-57, P greater than 0-1). No evidence was found for a release of prolactin conditioned by events normally preceding milking. The results showed a correlation between the average concentration of prolactin released at milking each month and the average milk yield of the six goats (r equals 0-67, P smaller than 0-05). The physiological significance of this correlation is discussed. No correlation was found between the average concentration of prolactin released during early lactation and the average milk yield of the goats (r equals 0-36, P greater than 0-1).


Seasonal factors affecting the release of prolactin in goats in response to milking. An investigation has been carried out in goats of the possible seasonal factors which might influence the autumn decline in the average concentration of prolactin released at milking. Serial blood samples were taken during milking, once a month between August and December 1972, from groups of mated, non-mated and ovariectomized goats and also from goats maintained in summer light conditions (17 h light: 7 hr darkness). Since high levels of progesterone occur in the circulation during the major part of the oestrous cycle and pregnancy in the goat, the same sampling procedure was carried out earlier in the year (June-August) in animals which had been implanted with progesterone tablets. Although high levels of progesterone in the circulation inhibited the release of prolactin at milking, oestrus, mating and pregnancy were not found to be responsible for the fall in prolactin levels in the blood in the autumn. In each of these cases the concentration of prolactin released at milking decreased uniformly between August and October. Housing goats in summer lighting conditions, however, maintained the release of prolactin at milking at a consistently high level from August until December, but had no effect on the milk yield which declined in a manner similar to that of the control animals. It is concluded that daylength is the predominant factor governing the autumn decline in the concentration of prolactin released at milking and that the decline is not the major cause of the fall in milk production during late lactation in goats.


Plasma progesterone and its relationship to serum gonadotrophins in immature female rats. In immature female rats, low values for concentrations of plasma progesterone were generally found from days 6-15 and from days 25-32 of life. Maximum progesterone concentrations (13-0-14-1 ng/ml), comparable to metoestrous values in the adult rat, occurred on days 20-22. The progesterone appeared to be of ovarian origin since after ovariectomy, on day 18, low progesterone concentrations were found 1 and 2 days later (2-5 ng/ml and 1-3 ng/ml) as compared with control values of 10-7 ng/ml and 14-1 ng/ml. However, adrenalectomy also lowered progesterone concentrations, 1 and 2 days later (6-4 and 4-9 ng/ml). The effect of progesterone, either alone or in combination with oestradiol benzoate (OB), on serum gonadotrophins was studied in rats ovariectomized on day 18. The highest dose of progesterone (0-15 mg) only slightly diminished the rise in serum luteinizing hormone (LH) after ovariectomy and had no effect on serum follicle-stimulating hormone FSH). Oestradiol benzoate in a dose of 0-025 mug/100 g body weight was highly effective in preventing the rise in both LH and FSH concentrations, and OB treatment (resulting in a near-physiological oestradiol concentration) combined with progesterone treatment was more effective than treatment with OB alone. The results suggest that the amounts of progesterone and oestradiol present in the 20-day-old rat are adequate to cause the decrease in FSH level normally observed in immature female rats around this age.


Effect of serotonin and progesterone on induced ovulation in immature rats. Serotonin (50 and 100 mg/kg), given subcutaneously, inhibited induced ovulation in immature rats treated with pregnant mare serum gonadotrophin (PMSG). A single injection was effective if given 52-55 h after the PMSG, i.e. in the 3 h period after the critical period before ovulation. The injection of serotonin inhibited the release of luteinizing hormone (LH) from the pituitary since the pituitary levels were higher than in the control animals and there was complete inhibition of the plasma LH surge. The anti-ovulatory effect was reversed by administration of progesterone and endogenous plasma progesterone levels were reduced in the late evening after serontonin treatment. The site of action of serotonin appeared to be peripheral since it inhibited induced ovulation in hypophysectomized rats but was without effect in intact rats treated intraventricularly. It is suggested that progesterone is essential for the occurrence of induced ovulation and serotonin inhibits either its secretion at the ovarian level or its passage away from the ovary.


Thyrotrophin concentrations in the plasma of cattle, sheep and foetal lambs as measured by radioimmunoassay. A radioimmunoassay technique for measuring the thyrotrophin (TSH) concentration of sheep and cattle plasma is described. The sensitivity of the assay allowed the measurement of 1-50 ng TSH/ml unextracted plasma. Cross-reaction with ovine luteinizing hormone, prolactin and growth hormone was very low. The average recovery of added TSH was 103 plus or minus 4-1 (S.E.M.)% and the between-assay coefficient of variation was 13-8%. The normal plasma TSH levels of sheep and cattle were approximately 2-5 ng/ml (5 mu. bovine TSH/100ml). Foetal sheep had plasma TSH concentrations of approximately 3-2 ng/ml during the last 20 days of gestation. Levels of TSH in the circulation decreased abruptly after hypophysectomy of the foetal lamb and a decline in the plasma thyroxine (T4) concentrations was also apparent within 24 h of the operation. However, thyroidectomy of adult and foetal sheep did not increase plasma TSH concentrations until almost all the T4 had been cleared from the circulation. The injection of T4 into thyroidectomized sheep rapidly reduced plasma TSH concentrations to normal values. However, the continued injection of T4 did not further reduce TSH concentration. The injection of T4 or triiodothyronine into normal sheep was also without effect on plasma TSH concentrations.


Animal psychophysics: improvements in the tracking method. It is difficult to maintain stimulus control in animal psychophysical studies using the tracking method. Loss of stimulus control is characterized by wandering thresholds and responses in the absence of the stimulus. Rats were trained to make a variable number of licking responses to turn on an auditory stimulus. A response on a lever in the presence of the stimulus was reinforced with food. Two precedures were added to improve stimulus control. First, lever responses in the absence of the stimulus raised the intensity of the stimulus at the next presentation; second, rate of reinforcement in the threshold region was maintained at about the same level as that in the suprathreshold region by reducing the number of licking responses required to turn on the stimulus. Using these two procedures, stimulus control was improved and maintained and reliable auditory intensity thresholds were obtained.


Some effects of response-independent reinforcement on auditory generalization gradients. Two groups of six rats received discrimination training with two auditory stimuli differing in intensity. During one stimulus, the schedule was variable interval; during the other, it was either variable time or extinction. Both the variable time and extinction schedules resulted in differential rates of responding in the presence of the two stimuli. Extinction resulted in an earlier and more stable difference. Stimulus generalization gradients obtained along the noise-intensity dimension revealed peak shift with both procedures. In addition, a secondary peak to stimuli in between the two training stimuli occurred with the variable-time schedule.


The effect of azathioprine (Imuran) on the cell cycle of promonocytes and the production of monocytes in the bone marrow. The present communication concerns the effect of azathioprine on the mitotic activity of promonocytes and the production of monocytes. In vitro and in vivo labeling with [3H]thymidine showed that during azathioprine treatment the promonocytes synthesize DNA and that, contrary to expectation, the labeling index increases. Cytospectrophotometric determination of the Feulgen-DNA content of the promonocytes during azathioprine treatment showed an increase in the percentage of tetraploid promonocytes, and determination of the various phases of the cell cycle showed significantly increased DNA synthesis and cell cycle times as compared with the normal steady state. On the basis of these results it can be concluded that azathioprine arrests the cell cycle of the promonocytes late in the DNA synthesis phase or in the postsynthesis (G2) phase and mitosis does not occur. This timing of the effect of azathioprine had not been previously observed. The diminished mitotic activity of the promonocytes during azathioprine treatment depressed monocyte production. During treatment with 3 mg/kg azathioprine the cell cycle time of the promonocytes was on the average 5.5 h longer than in the normal steady state and the rate of monocyte production was reduced by 70%. During an acute inflammatory reaction too, monocyte production is affected by azathioprine. In animals not treated with azathioprine but with an acute inflammation the cell cycle time becomes shorter and the monocyte production increases, but animals treated with (3 mg/kg) azathioprine do not show this effect. The kinetics of the monocyte also changes under the low dosage of azathioprine. As consequence of the diminished production of monocytes, far fewer (about 50%) monocytes enter and leave the circulation than during the normal steady state. During an acute inflammatory reaction the numbers in transit through the circulation are slightly augmented but remain considerably lower than in nonazathioprine-trehat of animals not treated with azathioprine.


Augmentation of delayed-type hypersensitivity by doses of cyclophosphamide which do not affect antibody responses. Mice immunized with more SRBC than are required to produce optimal delayed-type hypersensitivity reactions, developed good antibody responses and poor delayed foot pad reactions. Cyclophosphamide treatment in low doses (20 mg/kg) before immunization, augmented the delayed-type hypersensitivity without affecting antibody responses. Cyclophosphamide did not augment delayed responses to optimal doses of SRBC (0.01%), but did augment the delayed hypersensitivity response of mice immunized with a suboptimal antigen dose (0.001%); which produced no detectable antibody response with or without cyclophosphamide pretreatment. These results suggest that antibody feedback is not the sole regulator of delayed reactions; the possibility that suppressor T cells may also be involved is discussed.


Firearm residue detection by instrumental neutron activation analysis. Work leading to an improved technique for firearm discharge residue detection by neutron activation is described. The troublesome and time-consuming postirradiation chemistry has been eliminated; also, sample size has been minimized to accommodate some 130 samples per irradiation capsule. To put the method in a proper perspective, previous work has been referenced and discussed.


Change in muzzle velocity due to freezing and water immersion of .22, long rifle, K.F. cartridges. A study of change in muzzle velocity due to freezing and water immersion of .22, long rifle, K. F. cartridges has been presented. A statistical criterion has been formulated to ascertain whether or not a cartridge undergoes a change in muzzle velocity due to a particular treatment. The muzzle velocity data of .22, long rifle, K. F. cartridges, obtained by an electronic timer before and after the various treatments, have been analyzed in the light of this criterion. These cartridges have generally been found to suffer considerable loss in muzzle velocity when immersed in water for three weeks and also when immersed in water for three days and simultaneously cooled to 0 degrees C. The forensic significance of this loss in muzzle velocity has been discussed.


A portable explosives identification kit for field use. This study has shown the feasibility of using a miniaturized TLC kit for the presumptive identification of explosives in the field. The process of TLC field screening of a post-explosion site is nondestructive, resolves constituent explosives from interfering backgrounds, and increases the residence time of isolated specimens in the adsorbed state for subsequent substantive identification by trace methods of instrumental analyses. The cost of the modified Kodak kit for explosives identification (excluding the microscope) is approximately +150. The stock items, such as the fluorescent silica gel sheets, Cab-O-Sil, and solvents, are of nominal cost. The kit is relatively simple to use and personnel with a minimal technical background can be trained in its use.


Human or not? A problem in skeletal identification. Frequently, bones submitted to physical anthropologists by law emforcement agencies as human prove to be nonhuman. This results in a waste of time, money, and effort of all concerned. A course has been designed that could be offered by a physical anthropologist either as a workshop or as part of a training program for law enforcement officers. This would consist of teaching through lectures and laboratory work the techniques utilized by physical anthropologists to differentiate human from nonhuman bone, the anatomical relationships of bones of the human skeleton, and methods of recording bones and artifacts as they are recovered so that their original positions are documented both in relationship to each other and to the terrain. Training in these techniques would aid forensic investigators and amplify the kinds of information available to physical anthropological consultants asked to identify skeletal remains recovered by members of a law enforcement agency.


Is the insanity defense "unconscionable"? The debate over the meaningfulness and utility of the insanity defense has continued unabated ober the years. President Nixon has referred to the "unconscionably abuse" of the defense. This paper, presented as part of a panel on the subject, has propounded the view that the defense is unconscionable, using that aspect of the definition dealing with unreasonableness. The historical antecedents and the religious and social philosophy of the concept of responsibility and nonresponsibility have been reviewed. In addition to the inapplicability of the concept to current social problems, and the difficulties of applying current psychiatric knowledge to effect a rational delineation between the two legal entities encompassed under the rubric of responsibility and nonresponsibility, the potential problems and the potential opportunities which may result from the abolition of the plea are presented. With these factors in mind, as well as the obvious failure of the legal-social-penal system in handling the problems of the behaviorally deviant, I believe that the use of the current system has hampered the development of possibly more reasonable alternative systems and that, therefore, the maintenance of the insanity defense is unreasonable and harmful to our society. The insanity defense, as currently constituted and institutionalized, has evolved into a rigid and archaic vestige of the legal system of an earlier era and therefore its use has, in the sense defined, indeed become "unconscionable" and lacking in social meaningfulness.


Identification of drugs and other toxic compounds from their ultraviolet spectra. Part II: Ultraviolet absorption properties of thirteen structural groups. The ultraviolet absorption spectra of 13 different chemical classes of drugs and toxic organic compounds were studied. A classification system has been developed in which compounds with the same conjugated molecular system and auxochrome substituents are grouped together. Each of these groups has characteristic absorption spectra, showing similarities in the number of major bands, position of maximum absorbance, pH effects, and solvent effects. The absorption maxima and molecular absorptivities are tabulated for approximately 100 compounds, and characteristic spectra of each designated group are illustrated. Classes of drugs included in this study are pyridine derivatives, hydrazines, pyridylamine derivatives, variously substituted phenols, barbiturates, ureides, imides, hydantoins, and conjugted ketones (enones).


Sexual asphyxia in the female.

Sexual asphyxia in the female. The author has reported what appears to be a clearly defined masochistic hanging in the female. This is supported by many of the usual characteristics inherent in most male hangings. Although it was not the function of the police, a background study of the victim by a medical authority unfortunately was not made, which is so often the case in deaths due to sexual asphyxia.


Narcotic abuse among homicide victims in Detroit. A record of 751 homicides occurred in Detroit in 1973. An attempt was made to explore the relationship between narcotics and a random sample of these homicides. It was demonstrated that in the group studied, 43% were narcotic users. It is suggested that if one includes pushers and dealers (possibly victims of a "drug war"), people killed by addicts, and those cases where a narcotic might not be detected in the biological sample, the percentage of homicide victims associated with narcotics traffic could be as high as 60 to 70%. An investigation into the manner (executed, killed in a robbery attempt, etc) helps substantiate this suggestion. Finally, unlike the majority of homicide victims, ethanol is infrequently found in the victim using narcotics.


Carisoprodol-related death in a child.

Carisoprodol-related death in a child. A child who ingested approximately 3500 mg of carisoprodol gradually deteriorated and died within 36 h. GC analysis of serum, urine, and gastric samples indicated that meprobamate was the principal metabolite of carisoprodol.


Experience in the identification of abuse drugs in urines collected under Treatment Alternatives to Street Crime. This paper describes the criteria and analytical approach that were employed to cope with a mass drug urine screening program. Motives and justifications for selecting one technique over another are covered, along with a detailed description of those methods selected. Advantages, disadvantages, and limitations of each procedure are discussed. Results are presented for over 11,500 samples of urine collected from the criminal segment of Philadelphia's population over a period of six months. These were examined for amphetamine, methamphetamine, amobarbital, butabarbital, pentobarbital, phenobarbital, secobarbital, cocaine, codeine, methadone, and morphine.


Quantitative extraction of tubocurarine, gallamine, and decamethonium from biological materials. A procedure is described for quantitative extraction of tubocurarine, gallamine, and decamethonium from blood, urine, and tissue. Ion pairs of the drugs are formed with picric acid and are extracted into a dichloromethane/1-pentanol solvent. Qualitative analysis of the extract is effected using thin-layer chromatography. Quantitative analysis is accomplished by elution of TLC areas containing the drug, formation of the ion pair with sodium 9,10-dimethoxyanthracene-2-sulfonate, and analysis of the ion pair by spectrophotofluorometry.


Slow PIII component of the carp electroretinogram. The slow PIII component of the electroretinogram (ERG) was studied in the isolated, aspartate-treated carp retina. Although the latter is richly populated with cones, slow PIII appeared to reflect almost exclusively the activity of rods; e.g. the spectral sensitivity of the potential paralleled closely the rod pigment curve, its operating range (i.e. the V-log I curve) was limited to 3 log units above absolute threshold, and raising background intensities to photopic levels produced saturation of the increment threshold function without evidence of a cone-mediated segment. Only after bleaching away a significant fraction of the porphyropsin was it possible to unmask a small photopic contribution to slow PIII, as evidenced by a displacement in the action spectrum to longer wavelengths. The spatial distribution of the slow PIII voltage within the retina (Faber, D.S. 1969. Ph.D. Thesis. State University of New York. Buffalo, N.Y.; Witkovsky, P.J. Nelson, and H. Ripps. 1973. J. Gen Physiol. 61:401) and its ability to survive aspartate treatment indicate that this potential arises in the Müller (glial) fiber. Additional support for this conclusion is provided by the slow rise time (several seconds) and long temporal integration (up to 40s) of the response. In many respects the properties of slow PIII resemble those of the c-wave, a pigment epithelial response also subserved by rod activity. On the other hand, the receptoral (fast PIII) and the b-wave components of the ERG behave quite differently. Unlike slow PIII, response saturation could not be induced, since both potentials are subserved by cones when the stimulus conditions exceed the limits of the scotopic range. Receptors appear to govern light adaptation at photopic background levels; both fast PIII and b-wave manifest identical incremental threshold values over this range of intensities. However, under scotopic conditions, the sensitivity of the b-wave is affected by luminous backgrounds too weak to alter fast PIII threshold, indicating a postreceptoral stage of adaptation.


Sensitivity of calcium efflux from squid axons to changes in membrane potential. Squid giant axons were internally dialyzed with a medium free of metabolic substrates but containing 45Ca buffered with EGTA to concentrations of free Ca++ in the range 0.01-230 muM. At (Ca)i of 1.0 muM OR GREATER, Ca efflux was in the range of 1-3 pmol/cm2 s, was dependent on (Na)o and (Ca)o, and was sensitive to membrane potential. At lower (Ca)i, the sensitivity of Ca efflux to membrane potential was greater. Hyperpolarization of the membrane increased, and depolarization decreased Ca efflux over the range of potentials studied (-20 to -100 mV). The maximum sensitivity of Ca efflux to membrane potential was of the order of an e-fold increase in Ca efflux for a 25-mV increase in Em; this sensitivity of Ca efflux to membrane potential was lost if (Na)o was removed and was greatly reduced when (Ca)i was increased to 230 muM.


Olfactory receptor responses to sex pheromone components in the redbanded leafroller moth. Electrical responses of single olfactory receptor neurons of the male redbanded leafroller moth were elicited by each of the principle components of the sex pheromone and six other behaviorally active compounds. Response frequencies to equal intensities of each of these compounds  and changes in response frequency with increasing amounts of any one compound, varied from receptor to receptor. These differences in response characteristics appear to be due to factors intrinsic to the olfactory recptor neuron and not to factors external to it. The encoding of odor quality by these receptor neurons cannot be in the simple presence or absence  of activity in any one of them. Rather, odor quality may be encoded by the pattern of activity which invariably arises across an ensemble of receptor neurons, each having its own distribution of sensitivities.


Membrane response to current pulses in spheroidal aggregates of embryonic heart cells. Hearts from chick embryos aged 4,7, or 14 days were dissociated into their component cells, and the cells allowed to reassociate in the form of smooth-surfaced spheroidal aggregates on a gyratory shaker. Records from intracellular electrodes inserted into two widely spaced cells in a spontaneously beating aggregate indicated that the action potentials occurred virtually simultaneously. In aggregates made quiescent with tetrodotoxin, the voltage response to a current pulse injected in one cell could be noted by recording with a second microelectrode at various distance from the current source. The magnitude of the response was found not to vary with distance. It is concluded that the component cells in an aggregate are normally tightly coupled electrically; the cell boundaries do not constitute an appreciable resistive barrier. Such ag-regates behave as virtually isopential systems, with properties similar to those of single spherical cells, as modeled by Eisenberg and Engel (1970. J. Gen. Physiol. 55:736-757). Passive membrane time constant ranged from 11 to 31 ms, with a mean value of 17 ms; this value did not vary with aggregate size. Input resistance (V/I) varied inversely with aggregate size, as predicted, but with much scatter in the measured values. Specific membrane resistance was calculated as either 13,000 or 800 ohm-cm2 depending on whether input resistance was attributed to the total cell surface membrane area or to the outer surface of the sphere alone. No systematic difference in passive electrical properties of aggregates composed of 4-, 7-, and 14-day cells was seen. It is concluded that these aggregates may be suitable for voltage clamp analysis of their excitable membrane properties.


Metabolism of acetylcholine in the nervous system of Aplysia californica. I. Source of choline and its uptake by intact nervous tissue. Although acetylcholine is a major neurotransmitter in Aplysia, labeling studies with methionine and serine showed that little choline was synthesized by nervous tissue and indicated that the choline required for the synthesis of acetylcholine must be derived exogenously. Aanglia in the central nervous system (abdominal, cerebral, and pleuropedals) all took up about 0.5 nmol of choline per hour at 9 muM, the concentration of choline we found in hemolymph. This rate was more than two orders of magnitude greater than that of synthesis from the labeled precursors. Ganglia accumulated choline by a process which has two kinetic components, one with a Michaelis constant between 2-8 muM. The other component was not saturated at 420 muM. Presumably the process with the high affinity functions to supply choline for synthesis of transmitter, since the efficiency of conversion to acetylcholine was maximal in the range of external concentrations found in hemolymph.


Metabolism of acetylcholine in the nervous system of Aplysia californica. II. Reginal localization and characterization of choline uptake. The choline required for synthesis of acetylcholine is derived exogenously by  Aplysia ganglia. Under physiological conditions choline was taken up primarlily by neuropile and nerves and not by cholinergic cell bodies. In addition, compared with their contents of choline acetyltransferase, those components of nervous tissue which contain nerve terminals and axons synthesized acetylcholine far more efficiently. Choline was accumulated by high and low affinity uptake processes; the high affinity process appeared to be characteristic of cholinergic nuerons (Swartz, J. H., M. L. Eisenstadt, and H. Cedar.1975. J. Gen. Physiol. 65:255).  The two uptake processes were similarly affected by  temperature with a  Q10 of 2.8. Both were dependent on a variety of ions in a complicated manner. High affinity uptake seemed to be more dependent on Na+, showed greater inhibition by ouabain, and was selectively inhibited by oxotremorine. We found that the functional state  of neurons did not alter uptake of radioactive choline by either process, nor did it change the conversion to radioactive acetylcholine.


Metabolism of acetylcholine in the nervous system of Aplysia californica. III. Studies of an indentified cholinergic neuron. [3H] choline and [3H] acetyl CoA were injected into the cell body of an identified cholinergic neuron, the giant R2 of the Aplysia abdominal ganglion, and the fate and distribution of the radioactivity studied. Direct eveidence was obtained that the availabliity of choline to the enzymatic machinery limits synthesis. [3H] choline injected intrasomatically was converted to acetylcholine far more efficiently than choline taken up into the cell body from the bath. Synthesis from injected [3H] acety CoA was increased more than an order of magnitude when the cosubstrate was injected together with a saturating amount of unlabeled choline. In order to study the kinetics of acetylcholine synthesis in the living neuron, we injected [3H] choline in amounts resulting in a range of intracellular concentrations of about four orders of magnitude. The maximal velocity was 300 pmol of acetylcholine/cell/h and the Michaelis constant was 5.9 mM [3H] choline; these values agreed well with those previously reported for choline acetyltransferase assayed in extracts of Aplysia nervous tissue. [3H] acetylcholine turned over within the injected neuron with a half-life of about 9 h. The ultimate product formed was betaine. Subcellular distribution of [3H] acetylcholine was studied using differential and gradient centrifuagtion, gel filtration, and  passage through cellulose acetate filters. A small portion of acetylcholine was contained in particulates the size and density expected of cholinergic vesicles.


Relationships between voltage and tension in sheep cardiac Purkinje fibers. The two-microelectrode technique of voltage clamping sheep cardiac Purkinje fibers was used to examine the changes in contraction which occur during trains of voltage clamps. (A "train" is defined as a series of voltage clamps delivered at a particular rate, beginning after a rest long enough that the effects of previous stimulation have died away.) Contractions showed striking staircases, or progressive changes in peak isometric tension, during trains. Short clamps, clamps to voltages more negative than --20 or --30 mV, or holding potentials less negative than the resting potential favored negative staircases, while long clamps, clamps to positive voltages, and holding potentials near the resting potential each favored positive staircases. The staircase behavior appeared to be due to changes in the initial rate of recovery of the ability to contract. The changes in staircase behavior as a function of clamp voltage suggested that the relationship between peak tension and clamp voltage should depend on the experimental design. When the steady-state contraction was plotted as a function of clamp voltage, voltage-tension relations like those recently reported for working ventricle were obtained, with a threshold between --30 and --40 mV and a steep relation between tension and voltage. When the first contraction after a rest was plotted, the threshold voltage was more negative, the curve was flatter, and the peak tensions at inside positive voltages were reduced.


Slow inward current and contraction of sheep cardiac Purkinje fibers. A "slow" inward current (Is) has been identified in ventricular muscle and Purkinje fibers of several mammalian species. The two-microelectrode voltage clamp technique is used to examine some of the relationships between Is and contraction of the sheep cardiac Purkinje fiber. "Tails" of inward current occurring on repolarization and extrapolation of Is recovery each show that the Is system may not inactivate completely during prolonged depolarization. The rate of recovery of Is after a depolarization is slow, and when a train of 300-ms clamps (frequency 1 s-1) is begun after a rest, Is is larger for the first clamp than it is  for succeedings clamps. For the first clamp after a rest, the thresholds for Is and tension are the same and there is a direct correlation between peak tension and peak Is for clamp voltages between threshold and minus 40 mV. After a clamp, however, the ability to contract recovers much more slowly than does Is. Therefore, since Is may occur under certain conditions without tension, the realtionship between Is and tension must be indirect. Calcium entering the cell via this current may replenish or augment an intracellular calcium pool.


Anticonvulsants in the treatment of aggression. A significant number of violent acts are committed by individuals in whom central nervous system instability can be demonstrated by special electroencephalographic (EEG) activation procedures utilizing alpha-chloralose as the activating agent. Furthermore, subcortical electrograms suggest that this instability is related to a circumscribed ictal phenomenon in the limbic system. The abruptness of the aggressive act, the fact that the behavior is so often out of character for the individual and inappropriate for the situation, as well as the confusion and partial amnesia which accompany these episodes lend clinical support for the ictal hypothesis. Some anticonvulsants not only block the activated abnormalities on the EEG but also lead to dramatic clinical improvement in those individuals showing repeated and frequent aggressive behavior. For instance, in one study 46.7 percent and 53.3 per cent of the patients demonstrated activated abnormalities on no drug and placebo, respectively. When these same patients were receiving chlorpormazine or trifluoperazine, the activation rates were 60.0 per cent and 73.3 per cent, respectively. On the other hand, when these same patients were placed on a regimen of chlordiazepoxide the activation rate was reduced to 20 per cent (p smaller than or equal to .01). Another study involved severely distrubed chronically hospitalized psychotic patients whose aggressive uncontrolled outbursts relegated then not only to a locked ward, but often to isolation rooms despite high doses of phenothiazines. A regimen of chlordiazepoxide and


Clinician attitudes toward the suicide attempter. The clinical and sociodemographic characteristics and dispositional outcome of 248 suicide attempters were studied in a general hospital emergency room. A mood adjective checklist was completed by evaluating psychiatric residents and three orthogonal factor analytically derived mood clusters were related to sociodemographic, clinical, and dispositional variables. Residents expressed warmth toward patients having low suicide risk and limited overall psychopathology. They felt anxious toward patients with high suicidal risk and significant psychopathology. Angry feelings were reported toward patients with high suicidal risk in the absence of recent precipitating events. Residents reported warmth toward patients admitted to private or mental health center vs. state hospital facilities and tended to devote more time to their clinical assessment of these patients. Feelings of anxiety and anger characterized the responses to state hospital admissions who were only briefly assessed. The findings have implications for the organization of clinical services and the training of mental health professionals. Supervision needs to be directed to the recognition and modification of therapist emotional attitudes as well as to the understanding of patient psychopathology. Clinician feelings can then be effectively utilized to "capture" a high risk, high drop-out population into effective follow-up care.


Autonomic control, selective attention and schizophrenic subtype. Groups of paranoid reactive and nonparanoid process schizophrenic subjects participated in an experiment in which they were reinforced to modify attentional and autonomic responsiveness to differential excitatory and inhibitory avoidance stimuli. Groups did not differ in autonomic responsiveness or in ability to modify GSR and FPV activity to the differential avoidance stimuli. Groups were found to differ in their response to experimental instructions. Parenoid subjects reported being able to exert considerable emotional control during emotional stress. Nonparanoid subjects reported an inability to exert emotional control during stressful stimulation.


Increasing speech intensity of chronic patients ("mumblers") bu shaping techniques. The present experiment sought to increase the volume of speech in three adult male institutionalized patients. The method consisted of shaping modeled responses. Reinforcement was contingent upon the vocalization surpassing the threshold on an electronic volume indicator, which could be set at various levels of sensitivity. Results indicated that the volume of speech was increased in all three cases, as a function of manipulation of reinforcement contingencies. Follow-up data indicated that the long term maintenance of the acquired behavior was dependent on the ward environment.


Student suicide: death as a life style.

Student suicide: death as a life style. This is a study of the psychology of college students who are seriously suicidal. Fifty college students who had made suicide attempts were studied in detail over a 5-year period. Short term therapy was administered in most cases and long term therapy in some. Psychological tests (Wechsler Adult Intelligence Scale, Rorschach, Thematic Apperception Test, Sentence Completion, Figure Drawing, and Word Association) were performed and used as an independent check on the data derived from the interviews. The study indicates the ways in which death has become a way of life for these students--an integral, ongoing part of their adaptation. It traces the origin of this adaptation in a family relationship that the students perceived as requiring their emotional extinction. These students are tied to their parents in a kind of death knot and have become overly suicidal when live--coming to college, graduating, becoming seriously involved with another person--threatens to unravel this knot.


The family histories of manic-depressive patients with and without alcoholism. An investigation of family histories of bipolar affective disorder patients with and without an additional diagnosis of alcoholism showed that affective disorder was seen with equal frequency in both populations of relatives. Alcoholism was significantly more prevalent in relatives of the alcoholic probands than in the relatives of those who did not themselves have alcoholism. These data suggest that bipolar affective disorder and alcoholism may be transmitted independent of one another.


A wasted hand. Case with uncommon neurological and radiological features caused by a cervical band. The symptomatology in the thoracic outlet syndrome is well known. A patient is reported in whom the finding of a unilateral pulse deficit was the alerting sign that led to the correct diagnosis. Angiography, performed with injections during different respiratory phases, visualized the pathogenetic mechanism underlying some of the clinical findings.


Ictal laughter: a case report with clinical, cinefilm, and EEG observations. Seizures with laughter, verbalization, and repetitive abduction-adduction movements of the upper limbs are described in a 13 year old male of normal intelligence. Cinéfilm and EEG records illustrate some of the points made in the text.


A regional technique for the study of sensitivity to curare in human muscle. A regional technique for the study of curare sensitivity of human muslce in vivo is described. 0-5 mg d-tubocurarine is given intravenously at the wrist while the circulation to the hand and forearm is occluded. Neuromuscular transsmission is then studied by delivering trains of stimuli to the ulnar nerve and recording changes in evoked muscle action potential (map) amplitude from the abductor digiti minimi. In most normal subjects, this dose causes a recognizable block in neuromuscular transmission. The amplitude of a single evoked MAP is depressed and declines further during trains of 3/s repetitive stimulation. Recovery usually takes place gradually during the 20-30 minutes after the release of the tourniquet. The technique will be of value in the study of latent disturbance of neuromuscular transmission in neurological and metabolic disorders.


A study of sensitivity to curare in myasthenic disorders using a regionaltechnique. A study of curare sensitivity has been made in ocular myasthenia, myasthenia gravis, and the myasthenic syndrome sometimes associated with carcinoma, using a regional technique. Sensitivity is greater than in normal subjects. The injection of 0-125 mg d-tubocurarine resulted in a neuromuscular transmission block in all but a few of those cases with ocular myasthenia and in all cases of generalized myasthenia. Furthermore, the block persisted for longer than in normal subjects after the release of the tourniquet. This sensitivity to curare was not directly related to myasthenic weakness. The reasons for this are discussed and an explanation for such curare sensitivity is suggested.


Non-invasive screening for surgical intracranial lesions. The value and reliability of the combined results of skull radiographs, electroencephalography, echoencephalography, isotope angiography, and brain scanning in 147 patients suspected of having an intracranial space occupying lesions are analysed. The overall accuracy of the technique was 79%. No false negatives were found. The advantages of adopting the system proposed by the authors in everyday clinical work is discussed.