Metabolic response in severely traumatized patients. Acute severe trauma induces alterations in glucose, protein and fat metabolism through neurohormonal mechanisms. This study investigated the temporal relationship between serum cortisol, insulin levels and metabolic changes in glucose, protein and fat following acute severe trauma. Thirty adult patients admitted with polytrauma were investigated after two to three days of admission compared with thirty normal adults. There were highly significant increases in fasting blood glucose, ketones, triglycerides, cortisol and insulin, and highly significant decreases in total proteins and albumin. The study speculate that post traumatic hyperglycemia, decreased total proteins and albumin are major metabolic changes following severe trauma. These changes were mediated through neurohormonal mechanism included mainly the increased blood cortisol level.


Schistosoma haematobium (Egyptian strain) transmission microscopy of normal and praziquantel treated worms. The ultrastructure of the tegument and subtegumental structures of S. haematobium worms was examined by TEM. The effect of praziquantel on residual worms which remained after a curative dose was also studied. The male S. haematobium worms were characterized by deep tortuous pits and sharp apically directed spines. Spherical and elliptical bodies were also observed. The esophageal tegument was described. The effect of praziquantel on the male worms revealed flattening of spines and their disappearance in some regions as well as disruption and vacuolization of the tegument in others.


Evaluation of adult Schistosoma mansoni and cercarial antigens in serodiagnosis of schistosomiasis using IHAT and ELISA. Examination of 200 bilharzial cases divided into four groups and 50 normal controls to evaluate the efficiency of adult Schistosoma mansoni and cercarial antigens in serodiagnosis of schistosomiasis using IHAT and ELISA revealed that IHA test showed high sensitivity that decreased with the progress of clinical status of the disease if adult worm antigen was utilized. Also IHAT adult antigen was more sensitive for serodiagnosis than with cercarial antigen, while ELISA is less sensitive using both cercarial and adult antigens.


Naturally occurring toxoplasma antibodies in serum and milk of lactating women. Serum and milk of lactating women were tested for toxoplasmosis using specific-IgG IFAT. Apparently healthy 70 women were selected: 54 from rural and 16 from urban areas. Serum and milk were simultaneously collected from each one. Sera were positive in 22 (31.4%) of the total 70; including 16 (29.6%) and 6 (37.5%) of rural and urban groups respectively. No statistical significant difference was found for positivity and titre levels between the two groups (P greater than 0.05). Milk was positive in 12 (17.1%) of the 70 women; including 10 (18.5%) and 2 (12.5%) from rural and urban groups respectively, having no statistical significant difference (P greater than 0.05). Comparing serum and milk for positivity and titre levels, also there was no statistical significant difference (P greater than 0.05). It is concluded that relatively low antibody levels in serum could be excreted in milk and may be protective for suckling babies. Occurrence of antibodies in serum and milk are homogeneously distributed between rural and urban inhabitants.


Sampling genotypes on complex pedigrees with phenotypic constraints: the origin of the B allele among the Polar Eskimos. Exact probability calculations are often infeasible on large complex pedigrees. Conditional independences, however, occurring as a natural consequence of Mendelian inheritance of genetic traits, define a locally dependent Markov random field on the state space of all genotypic configurations on the pedigree. The underlying Markov chain is irreducible for most traits determined by a diallelic locus. For a given pedigree and a known genetic model, the Gibbs sampler can be used to obtain good estimates of the posterior distribution of genotypes given the observed data. The areas of pedigree analysis to which such an approach would be most directly relevant include genetic counselling and selective animal breeding, together with questions about ancestral genotypes and the ancestral paths of rare alleles. The method is illustrated by tracing the ancestral paths of a rare allele in a simple diallelic system on a highly complex Eskimo pedigree.


The dynamics and control of bovine tuberculosis in possums. A model for the dynamics of an infectious disease in a wild animal population is analysed. The model incorporates susceptible, infected but not infectious, and infectious classes, with no recovery from disease, density-dependent regulation of host birth and death rates, and pseudo-vertical transmission. The existence and local stability of equilibrium levels are determined. Expressions for the intensity of three alternative control measures - the culling, sterilization, and vaccination of host animals - required to eradicate the disease are obtained. Finally, the model is applied to possible control measures against bovine tuberculosis infection in possums in New Zealand.


The population dynamics of nematode infections of ruminants: the effect of seasonality in the free-living stages. A previously published differential equation model for the dynamics of nematode infections of ruminants is modified to allow for time-dependent development and loss rates in the free-living stages. This converts the nonlinear autonomous system to one with periodic coefficients. Fourier transform methods are used to analyse the response of the modified system to these forced oscillations. For biologically reasonable parameter values, the annual pattern of parasitism is determined for the nonautonomous system, and compared with that for the autonomous system with periodic perturbations, and a system incorporating both effects. It is found that, whereas periodic perturbations due to management intervention determine the qualitative annual patterns of larval abundance and host infection, the seasonal dynamics of the larval stages change the magnitude of the adult worm burden.


[Clinical significance of direct immunobead test to detect anti-sperm antibody]. Direct immunobead test (IBT) (IgG.IgA) was carried out for 290 infertile men to detect antisperm antibodies. The results, compared with semen data analyzed by traditional methods and a computer assisted semen analyzer and with hormone levels, were as follows. 1) In 21(7.2%) out of the 290 men, IgG-immunobead (IB) binding was observed in more than 50% of their motile spermatozoa; in 10(3.4%) of the 290, IgA-IB binding. 2) In all of the cases with proven fertility, the IB binding rate was less than 60% for IgG-IB, and less than 40% for IgA-IB. This suggested that antibodies attached to sperm could be among the factors in male infertility. 3) Antibodies attached to the sperm head with binding rate of less than 40% were not considered to be a causative factor in male infertility. 4) The incidence of positive IgG-IB was higher, and also in each case the IgG-IB binding rate was higher than the IgA-IB. 5) IgG-IB binding parts of each spermatozoon included IgA-IB binding parts. 6) No significant relationship was observed between the results of traditional semen analysis, LH.FSH.PRL levels or PENETRAK and the rates of IB binding.


[Evaluation of myometrial invasion in patients with endometrial cancer by magnetic resonance imaging]. Magnetic Resonance Imaging (MRI) has been widely used for the diagnosis of gynecologic disease. We studied the clinical usefulness of MRI in 34 patients with endometrial cancer from July, 1988 to August, 1991. Pre-operative pelvic MRI was carried out to evaluate the myometrial invasion in these patients. MRI was accurate in predicting the presence or absence of myometrial invasion in 23 of 34 cases (68%), while echo or CT (computed tomography) was accurate in 26% and 8%. In conclusion, MRI is considered to be the most useful examination for the evaluation of myometrial invasion in endometrial cancer.


[A group study on prognosis of ovarian cancer in Japan]. An assessment has been made, with the cooperation of 22 institutes, of 1,185 cases of ovarian cancer as subjects who were treated in the period from January, 1980, to December, 1987. As a result, (1) As for distribution by clinical staging at the initial examination, the cases in Stage III were the most numerous, followed by those in Stage I, and if classified according to the histological type, serous cystadenocarcinoma was the most frequently observed in Stage III, and undifferentiated and unclassified carcinomata were observed in Stages III and IV. (2) In the examination of prognostic factors, it was confirmed that the clinical stage, histological type and diameter of the remaining tumor after the initial operation were important factors. (3) A significant difference was observed between the grade of histomorphological differentiation and prognosis, the difference was chiefly due to the deviated distribution of clinical staging in each group of differentiation. (4) A favorable trend was observed in the prognosis by patient's age toward the younger layer. (5) When the starting time of the therapy is considered, a trend toward improvement has been seen year by year, and it is considered that the beneficial effect of chemotherapy with CDDP contributes to this.


[Effect of suppletory estrogen or 1,25(OH)D3 on bone mineral content]. Post-menopausal changes in bone mineral content (BMC) and the effect of suppletory estrogen or 1,25(OH)D3 was studied. BMC was evaluated by means of quantitative computed tomography. Post-menopausal BMC (age 45-49) was 158.4 +/- 42.0 mg/cm3 which was significantly lower than in other subjects in the same aged group (192.3 +/- 27.1 mg/cm3). At two years, ten years and more than ten years post-menopause it decreased significantly by 17%, 30%, 55% respectively, compared with that of pre-menopausal BMC. The administration of conjugated estrogen (0.625 mg) or 1,25(OH)D3 was effective in preventing bone loss, although a transient decrease in BMC was observed in some cases to which 1,25(OH)D3 was administered. In cases of pre-menopausal bilateral oophorectomy, BMC was found to decrease significantly to the post-menopausal level about two years after operation. However, long-term estrogen replacement therapy tends to inhibit the decrease.


[A trial of low-dose aspirin therapy in high-risk pregnancy]. Intra-uterine growth retardation, intra-uterine fetal death and pre-eclampsia have common abnormalities: A reduction of uteroplacental perfusion, lack of vasodilation of spiral arteries and subsequent thrombosis. These physiological processes have been explained by an imbalance between prostacyclin and thromboxane A2 production. Many studies have suggested that treatment with low-dose aspirin and steroids is effective in preventing pregnancy loss or pre-eclampsia, but the mechanism has not been established. We evaluated the effectiveness of these therapies in patients at risk for pregnancy loss with the aspect of intracellular ionized calcium mobilization. Low-dose aspirin directs the prostacyclin/thromboxane A2 balance to the dominance of prostacyclin and steroids suppress the activities of lupus anticoagulant or antiphospholipid antibodies. The intracellular ionized calcium concentration in platelets is decreased significantly after these therapies. Concerning the pathological examination of placenta, there were deposits of fibrin in only 2 out of 8 cases and there were no abnormal findings in the other 6 cases. These data show that the aggregation of platelets is suppressed in microvascular circulations. These therapies do not cause any adverse effect on the mother or fetus. It is concluded that low-dose aspirin therapy with steroids is useful for patients with a poor obstetrical history.


[Morphological investigation in microscopic mole]. Although regulations for treating trophoblastic disease have been established, the diagnostic standards and biological behavior of the microscopic mole are not always described distinctly. I observed morphological changes in the various villi existing between abortion villi and normal villi or total mole and investigated problems related to the microscopic mole. As a result, (1) the disappearance of the villous vessels, central liquefaction and proliferation of the villous cells were observed, though at a low incidence, not only in patients with abortion but also in those with normal villi; (2) When the short axis of the villi was 1,000 microns or less, there were no patients in which villus vessels disappeared, or central liquefaction or proliferation of the villous cells occurred; (3) Among patients with villi having a short axis of 1,000 microns or more, there were patients in whom trophoblastic disease could not be completely distinguished from total mole. A successive tumor developed in 2 patients in the group with (3) findings; (4), I propose that patients who satisfy the criteria in (3), above, should be regarded as those with microscopic mole while others should be regarded as those with hydropic degeneration in a narrow sense.


[A study on relationship between functional sterility and ultrasonographic endometrial images]. By means of vaginal ultrasonography 38 cases with functional sterility were investigated. Ultrasonographic endometrial thickness and texture, along with the maximum follicle diameter, serum estradiol and progesterone levels were measured in connection with the outcome of treatment for sterility. Endometrial texture was classified into 4 types: even-hypoechoic (P1) and mixed (P2) in late proliferative and even-echogenic (S1) and mixed (S2) in mid-secretory phases. Histological findings were also compared with ultrasonographic images and the following results were obtained. 1. Endometrial thickness and outcome of sterility. 1) Endometrial thickness in mid-proliferative and mid-secretory phases was 9.7 +/- 1.4 mm (mean +/- SD) and 13.9 +/- 1.7, respectively, in a group with good prognosis, while it was 6.7 +/- 1.8 and 10.1 +/- 2.2 in a poor prognosis group. The endometrium was significantly thinner in the latter group than in the former (p less than 0.05). 2) No. difference was found in estradiol, progesterone levels or the maximum diameter of the follicle. 2. Endometrial texture and outcome of sterility. 1) The outcome was identical in both groups whether the endometrium in late proliferative phase was of the even-hypoechoic or of the mixed type. 2) In the mid-secretory phase, comparison of patients where endometrium was of the even-echogenic type (S1) with those who had the mixed type (S2), revealed clearly poorer prognosis in the latter (p less than 0.05). 3) Serum estrogen and progesterone levels were not significantly different in any endometrial patterns and no connection was found between endometrial textures and the hormonal levels. 3. Histological findings in cases where mixed pattern endometrium was observed.(ABSTRACT TRUNCATED AT 250 WORDS)


Lipopolysaccharide, tumor necrosis factor, and interleukin-1 interact to cause hypotension. Lipopolysaccharide (LPS) causes the syndrome of septic shock by initiating the release of endogenous mediators such as tumor necrosis factor (TNF) and interleukin-1 (IL-1) from macrophages. Hypotension is one of the important clinical features of septic shock; however, TNF is only hypotensive in high doses. Therefore we have investigated the interactions of low, nonhypotensive doses of LPS, IL-1, and TNF in the restrained unanesthetized rabbit. Combinations of nonhypotensive doses of TNF, IL-1, and LPS produced significant (p less than 0.05) decreases in blood pressure as compared with doses of each of the substances alone. TNF bioactivity in animals that were made hypotensive with combinations of TNF, IL-1, and LPS was lower than in animals that were made hypotensive with TNF alone. This suggests that TNF release that is stimulated by LPS is not the sole cause of the hypotension that is seen in this model of endotoxic shock. In this model, interactions of LPS, IL-1, and TNF occur and may explain hypotension during some episodes of sepsis.


Activated peripheral blood mononuclear cells in IgA nephropathy express platelet-derived growth factor B-chain messenger RNA. We investigated platelet-derived growth factor (PDGF) A- and B-chain messenger RNA expression in peripheral blood mononuclear cells or T cells obtained from 26 patients with IgA nephropathy and 15 healthy age-matched control subjects. Most patients with IgAN (88%) showed elevated PDGF B-chain expression in peripheral blood mononuclear cells; no PDGF B-chain expression was detected in peripheral blood mononuclear cells of normal control subjects. In T cells from both patients with IgAN and normal control subjects, however, PDGF B-chain messenger RNA expression was not detected. A positive correlation was noted between PDGF B-chain messenger RNA levels and urinary protein excretion. The PDGF B-chain messenger RNA expression in peripheral blood mononuclear cells also correlated with histopathologic changes in renal tissue obtained from patients with IgAN. Sixty-five percent of patients with grade III or IV histopathologic findings showed strong PDGF B-chain gene expression in their peripheral blood mononuclear cells. We could not detect PDGF A-chain messenger RNA expression in either peripheral blood mononuclear cells and T cells obtained from patients with IgAN and from healthy control subjects. These studies suggest that abnormally regulated PDGF B-chain expression in peripheral blood mononuclear cells may be associated with the progression of IgAN and may be useful as an indicator of disease activity.


Thrombin induction of plasminogen activator inhibitor mRNA in human umbilical vein endothelial cells in culture. We have previously reported that incubation of human umbilical vein endothelial cells (HUVECs) with human alpha-thrombin causes a time- and concentration-dependent increase in secreted plasminogen activator inhibitor type 1 (PAI-1) activity (Gelehrter TD, Sznycer-Laszuk R. J Clin Invest 1986;77:165-9). Here we report that the regulation of PAI-1 activity by thrombin is secondary to the thrombin-induced increase in PAI-1 mRNA accumulation. Incubation of HUVECs for 6 to 24 hours with 0.3 to 1.0 U/ml thrombin causes a 1.8-fold to 10-fold increase in PAI-1 activity and a 1.5-fold to threefold increase in accumulation of both the 3.2 and the 2.2 kilobase PAI-1 mRNAs. These effects are prevented by the prior addition of hirudin, a specific thrombin inhibitor. Inhibition of RNA synthesis with actinomycin D blocks the thrombin induction of PAI-1 mRNA accumulation. The protein synthesis inhibitor, cycloheximide, which itself markedly stimulates the accumulation of PAI-1, appears to prevent the induction by thrombin, suggesting that thrombin may act by inducing another effector such as interleukin-1. Consistent with this hypothesis is our observation that simultaneous addition of antibodies to interleukin-1-alpha prevents the thrombin induction of PAI-1 activity and mRNA.


The role of cytokines in polymyositis: interferon-gamma induces class II and enhances class I major histocompatibility complex antigen expression on cultured human muscle cells. Aberrant expression of class II major histocompatibility complex molecules has been found on target cells of various autoimmune diseases, including muscle fibers in patients with polymyositis-dermatomyositis. In this study the effects of a number of recombinant human cytokines, individually and in combination, on class I and class II molecule expression by cultured human muscle cells were examined with monoclonal antibodies and an immunoperoxidase technique. The following cytokines were tested: interferon-gamma, tumor necrosis factor-alpha, tumor necrosis factor-beta, interleukin-2, interleukin-1 alpha and interleukin-1 beta. Only IFN-gamma induced expression of class II molecules in muscle cells. It also enhanced the preexisting class I molecule expression by muscle cells. These findings suggest that IFN-gamma is involved in the aberrant expression of major histocompatibility complex molecules in the affected muscles of patients with polymyositis-dermatomyositis.


Surfactant protein A in bronchoalveolar lavage fluid. We measured surfactant protein A and phosphatidylcholine in the bronchoalveolar lavage fluid from healthy volunteers and several groups of patients with lung diseases to obtain information on surfactant in the lung. We developed three types of enzyme-linked immunosorbent assays that used combinations of polyclonal antiserum and monoclonal antibodies. Phosphatidylcholine was assessed by enzymatic measurement. The median amounts of surfactant protein A in bronchoalveolar lavage fluid determined by the enzyme-linked immunosorbent assay with monoclonal antibodies were as follows: control subjects (n = 10), 2.82 mg/L (range, 0.92 to 5.17 mg/L); patients with asthma (n = 13), 1.89 mg/L (range, 0.45 to 2.95 mg/L); and patients with pulmonary sarcoidosis (n = 20), 2.98 mg/L (range, 0.68 to 7.02 mg/L). The median phosphatidylcholine concentrations in the bronchoalveolar lavage fluid were as follows: control subjects (n = 10), 20 mumol/L (range, 3 to 37 mumol/L); patients with asthma (n = 12), 24 mumol/L (range, 3 to 55 mumol/L); and patients with pulmonary sarcoidosis (n = 20), 26 mumol/L (range, 4 to 76 mumol/L). As a group, the patients with asthma had less surfactant protein A in bronchoalveolar lavage fluid than did the control subjects (Mann-Whitney U test, p less than 0.05). The surfactant protein A levels measured by the enzyme-linked immunosorbent assay with polyclonal antiserum and by a competitive enzyme-linked immunosorbent assay were also lower in bronchoalveolar lavage fluid from patients with asthma than in that from control subjects. The phosphatidylcholine concentrations in all groups were similar.(ABSTRACT TRUNCATED AT 250 WORDS)


Quantitative determination of porphyrins in rat and human urine and evaluation of urinary porphyrin profiles during mercury and lead exposures. Measurement of urinary porphyrin excretion patterns (porphyrin profiles) is useful in the diagnosis and evaluation of diseases and disorders of porphyrin metabolism. However, experimental investigation of such disorders with rodent models has been hampered by the lack of an efficient procedure for the isolation and quantitative evaluation of porphyrins in rodent urine. This article describes an analytic procedure that overcomes the principal difficulties encountered with determination of porphyrins in rodent urine, including the loss of porphyrins during their isolation and interference of porphyrin fluorescence by contaminating materials. The procedure entails application of an acidified urine sample to a preconditioned C-18 preparatory column, preferential separation of essentially all potentially interfering contaminants by sequential phosphate-methanol elution, and selective isolation of porphyrins, which are then separated and quantitated by high-performance liquid chromatography and spectrofluorometric techniques. This method has been used to characterize urinary porphyrin excretion patterns in male rats and to define the distinctive changes in porphyrin profiles associated with prolonged exposure to porphyrinogenic metals. The porphyrin excretion patterns of male and female human subjects are also described. This method is applicable to the investigation of urinary porphyrin profile changes associated with exposure to a wide range of porphyrinogenic chemicals in both animals and human subjects.


Dimethyl sulfoxide decreases lung neutrophil sequestration and lung leak. To investigate basic mechanisms of acute edematous lung injury (adult respiratory distress syndrome), the formylated tripeptide formyl-norleucyl-leucyl-phenylalanine (FNLP) was instilled intratracheally into hamsters. Intratracheal FNLP produced time-dependent and dose-dependent increases in neutrophils recoverable by lung lavage (neutrophil alveolitis) and leak of intravenously injected albumin into the extravascular lung space (lung leak). Treatment with dimethyl sulfoxide (DMSO) decreased (p less than 0.05) neutrophil alveolitis and lung leak in hamsters given FNLP intratracheally. The effect of DMSO on various neutrophil functions was also studied in vitro. Addition of DMSO at concentrations (about 0.20%) measured in plasma of hamsters given DMSO decreased (p less than 0.05) neutrophil chemotaxis but not neutrophil superoxide anion generation or adherence to cultured endothelial cell monolayers or nylon fiber in vitro. We conclude that intratracheal FNLP causes neutrophil alveolitis and lung leak and that DMSO treatment ameliorates these processes, possibly by inhibiting neutrophil chemotaxis.


Plasma histamine but not anaphylatoxin levels correlate with generalized urticaria from infusions of anti-lymphocyte monoclonal antibodies. Anti-lymphocyte monoclonal antibodies have shown promise in trials for therapy of lymphocyte malignancies but are associated with a high frequency of immediate-type anaphylactoid reactions. We have previously demonstrated that these immediate-type anaphylactoid reactions are not mediated by immunoglobulin E to anti-lymphocyte monoclonal antibodies. To gain insight into the mechanisms of these immediate-type anaphylactoid reactions, we measured plasma levels of histamine and anaphylatoxins (C3a, C4a, C5a) during 11 infusions in eight patients who received anti-lymphocyte monoclonal antibodies (T101 and Lym-1). Three patients experienced generalized urticaria (two with severe angioedema); a fourth patient had three isolated hives but without generalized manifestations of an immediate-type anaphylactoid reaction. Plasma histamine levels after infusions that were associated with generalized urticaria were significantly higher than those during infusions that were not associated with generalized urticaria (mean, 3.47 vs 0.18 ng/ml, p less than 0.001). Increases in C3a and C4a levels were observed after some infusions, but these did not correlate with generalized urticaria. Measurable rises in plasma C5a levels after infusions were not detected. Although these data should be viewed as preliminary considering the limited number of patients studied, the observed histamine release demonstrates that mast cell or basophil activation that is not mediated by immunoglobulin E to anti-lymphocyte monoclonal antibodies occurs in the pathogenesis of immediate-type anaphylactoid reactions from anti-lymphocyte monoclonal antibodies. Although activation of the classical complement pathway may occur in some anti-lymphocyte monoclonal antibody infusions, this does not appear to explain immediate-type anaphylactoid reactions.(ABSTRACT TRUNCATED AT 250 WORDS)


Effects of probucol in renal function and structure in rats with subtotal kidney ablation. Probucol is a bisphenolic compound that lowers serum cholesterol and also has potent antioxidant properties. The present studies examined the effects of probucol administration on renal function and structure in a rat model of subtotal renal ablation. After subtotal nephrectomy, rats were fed an isocaloric rat chow diet containing 22.8% protein with or without the addition of 1% probucol. After 4 weeks, clearance studies were performed for determination of glomerular filtration rate (inulin clearance) and effective renal plasma flow (paraaminohippurate clearance). After completion of clearance studies and measurements of arterial blood pressure, the animals were exsanguinated and renal tissue was obtained for histologic evaluation. There were no differences in body weight, hematocrit, and blood pressure between the two groups of rats 4 weeks after subtotal nephrectomy. Rats with a remnant kidney given probucol had a significantly lower serum cholesterol level (47.4 +/- 5.3 mg/dl vs 87.2 +/- 10.4 mg/dl) and urea nitrogen level (40.7 +/- 3.2 mg/dl vs 63.6 +/- 8.1 mg/dl) than the control group. Rats given probucol also had significantly greater values for inulin clearance and clearance of paraaminohippurate and significantly less proteinuria than control rats. Also, rats with a remnant kidney given probucol had a significantly greater number of normal glomeruli (6.2% +/- 2.1% vs 1.1% +/- 0.9%) and a lesser number of severely affected glomeruli, grades III and IV (26.0% +/- 5.9% vs 50.9% +/- 9.1%) than rats with a remnant kidney not given probucol. Tubulointerstitial changes also were significantly less in rats with a remnant kidney given probucol.(ABSTRACT TRUNCATED AT 250 WORDS)


Microplate and dot immunoassays for the serodiagnosis of tuberculosis. A simple dot enzyme immunoassay based on the recognition of serum IgG antibody to a 30,000 dalton native antigen purified from culture filtrates of Mycobacterium tuberculosis was developed and compared with a standard plate enzyme-linked immunosorbent assay in the serodiagnosis of tuberculosis. The previously described favorable test characteristics of plate enzyme-linked immunoassay were confirmed; although the dot enzyme immunoassay was promising, it was less satisfactory. Dot enzyme immunoassay may have its most promising use as a screening test for situations of limited technical facilities. Both plate enzyme-linked immunoassay and dot enzyme immunoassay had markedly reduced sensitivities in persons with human immunodeficiency virus infection.


The effects of inhibition of plasma cholinesterase and hepatic microsomal enzyme activity on cocaine, benzoylecgonine, ecgonine methyl ester, and norcocaine blood levels in pigs. We measured the blood levels of cocaine and its three major metabolites, benzoylecgonine, ecgonine methyl ester, and norcocaine, in three groups of male pigs weighing about 26 kg (25.75 +/- 0.25 kg) to determine the effects of inhibition of plasma cholinesterase and hepatic microsomal enzyme activity on cocaine metabolism. In addition, systemic elimination half-life, volume of distribution, and clearance of cocaine were calculated for the three groups. Group 1 pigs (n = 4) were pretreated with normal saline solution, group 2 pigs (n = 4) were pretreated with tetraisopropyl pyrophosphoramide, a specific plasma cholinesterase inhibitor, and group 3 pigs (n = 4) were pretreated with cimetidine, a hepatic microsomal enzyme inhibitor, all administered intramuscularly. Pigs were anesthetized with intravenous sodium thiopental; a carotid arterial cannula and an external jugular catheter were then inserted for the administration of cocaine and for blood sampling. Forty-five minutes later, when pigs were again completely awake, cocaine 3 mg/kg was given intravenously. Arterial blood samples were collected for the analysis of cocaine and cocaine metabolite levels just before and at 5, 10, 15, 30, 45, 60, 120, 180, and 1440 minutes after the administration of cocaine. Cocaine and cocaine metabolite blood levels were analyzed with high-pressure liquid chromatography methods and plasma cholinesterase activity was measured with a colorimetric method. The blood levels of cocaine and cocaine metabolites were significantly different among the three groups (p less than 0.05, analysis of variance). Statistically significant differences in half-life, volume of distribution and clearance were also seen among the three groups.(ABSTRACT TRUNCATED AT 250 WORDS)


IEC-18, a nontransformed small intestinal cell line for studying epithelial permeability. Small intestinal epithelium is leaky and allows permeation of hydrophilic molecules of various sizes. Passively absorbed hydrophilic permeability probes have been shown to permeate across intestinal epithelium mainly through the paracellular pathways. In this study we introduce microporous filter-grown IEC-18 epithelial cells, a nontransformed small intestinal cell line, as a in vitro model of intestinal epithelium for the study of epithelial permeability. IEC-18 cells, originally derived from native rat ileal crypts, form confluent epithelium when grown on hydrated collagen-coated Millicell-CM permeable inserts (Millipore Corp., Bedford, Mass.). With scanning and transmission electron microscopy, the presence of tight junctions and desmosomes between cells and the development of microvilli at the apical surface were confirmed. Immunofluorescent labeling of ZO-1 proteins and desmoplakins verified the presence of tight-junctional proteins (ZO-1) and desmosomes in the intercellular junctions of confluent IEC-18 epithelium. The net electrical resistance of IEC-18 epithelium (28 omega-cm2) was similar to resistance values obtained from small intestinal tissue with (50 to 100 omega-cm2) or without (20 to 45 omega-cm2) muscularis and serosal layers. Assessment of mannitol and dextran permeation revealed early "maturation" of paracellular pathway, with increasing restriction of permeation to both probes through day 4. Resistance across IEC-18 epithelium also reached plateau levels between 4 and 7 days. Permeability studies with various probes indicate that cross-sectional diameter rather than molecular weight of the probe is the important determinant of permeation rate. IEC-18 epithelium selectively restricted the permeation of probes proportional to probe size; permeation of larger probes such as albumin was negligible. We conclude that cultured IEC-18 epithelial cells, because of their native crypt origin, similarity in resistance to small intestinal epithelia, retention of ability to differentiate into villus-like enterocytes, and permeability characteristics, are a useful model of intestinal epithelium for the study of permeability and paracellular transport.


Pulmonary emphysema in a nonsmoker with normal alpha-1-antitrypsin. Premature and rapidly progressing emphysema of the lung is unique in the absence of cigarette smoking or an inherited homozygous alpha-1-antitrypsin deficiency. We describe a young man with radiographic evidence of early onset bullous emphysema without the usual risk factors and provide a review of the alternative explanations proposed for this form of lung damage.


DaCosta's syndrome: chronic symptomatic hyperventilation. In 1871, DaCosta published his observation of somatic symptoms preceded by significant hyperventilation. More than a century later, the hyperventilation syndrome remains a poorly defined but common clinical condition. Although familiar to most practitioners of medicine when it presents as an acute phenomenon, the diagnosis may go unrecognized in its chronic form. The ability of a chronic hyperventilatory state to mimic a life-threatening cardiopulmonary disease is not always appreciated.


Nonsteroidal anti-inflammatory drug induced renal syndromes. Nonsteroidal anti-inflammatory drugs (NSAIDs) may cause acute renal failure from unopposed vasoconstriction or acute interstitial nephritis. NSAID induced hemodynamic renal failure is characterized by sudden oliguria, often with decreased fractional excretion of sodium, occurring in patients with decreased effective circulating fluid volume or preexisting renal disease. Allergic interstitial nephritis from NSAIDs may occur at any time during therapy with the drugs and may present as renal failure with or without the nephrotic syndrome. Although chronic renal failure has been reported, both renal syndromes usually resolve when treatment with the NSAID is discontinued. Renal function should be measured soon after initiation of therapy in patients at risk for the hemodynamic effects of the drugs and periodically thereafter. Patients should be warned about the potential toxicity of the drugs.


Elevating maternal insulin-like growth factor-I in mice and rats alters the pattern of fetal growth by removing maternal constraint. Fetal growth is normally constrained by maternal factors. This constraint is demonstrated by the usual inverse linear relationship between litter size and mean fetal weight. Cross-breeding experiments between mice of lines selected for high or low plasma insulin-like growth factor (IGF-I) levels suggested that elevations in maternal IGF-I abolish (P less than 0.01) this constraining effect and reverse the usual positive relationship between fetal and placental size in late gestation. This was confirmed by treating mice and rats throughout pregnancy with IGF-I. In normal mice and in low IGF-I line mice treatment with IGF-I (10 micrograms 8-hourly s.c. from day 1 to 19 of pregnancy) abolished maternal constraint whereas 0.9% (w/v) NaCl treatment did not. In Wistar rats osmotic pumps were implanted to deliver IGF-I (1 microgram/g body weight per day), bovine GH (bGH; 0.6 microgram/g body weight per day) or saline from day 1 to 19 of pregnancy. IGF-I therapy but not bGH or saline abolished (P less than 0.01) maternal constraint and altered (P less than 0.01) the relationship between placental and fetal weight. When high or low IGF-I line mice embroys were transplanted into a normal line of mice, the expected negative relationship (P less than 0.05) between mean fetal weight and litter size was maintained. However, the embryos of the high line were heavier (P less than 0.05) than those from the low line irrespective of fetal number, suggesting a direct role for IGF-I in the regulation of fetal growth.(ABSTRACT TRUNCATED AT 250 WORDS)


A comparison of plasma vasopressin and oxytocin concentrations during the oestrous cycle of the ewe. Blood samples were collected twice daily (09.30 and 17.00 h) via indwelling jugular-vein cannulae from five ewes throughout an entire oestrous cycle. Additional frequent samples were taken at 30-min intervals for 8 h on each of days 3 (early luteal phase), 9 (midluteal phase), 12 (late luteal phase) and 0 (day of oestrus). Plasma concentrations of arginine vasopressin and oxytocin were measured in all samples by radioimmunoassay and progesterone was measured in the twice-daily samples only. Both oxytocin and progesterone showed the expected pattern of plasma concentrations, increasing during the early luteal phase, reaching a plateau and declining either preceding (oxytocin) or at (progesterone) luteolysis. Vasopressin concentrations showed a significant dependence on the day of cycle (P less than 0.05, analysis of variance) with concentrations lowest at oestrus and minor peaks on days 4 and 8-9. There was no correlation between the concentrations of vasopressin and progesterone. Vasopressin values were significantly higher in the morning than in the afternoon samples (1.3 cf. 0.9 pmol/l; P less than 0.001). Analysis of the frequent samples showed a significant (P less than 0.001) dependence on the time of day for vasopressin but not oxytocin. Values were high throughout the morning, declined to a trough at 15.00 h and rose again by 17.00 h. We conclude that there is a minor variation in the vasopressin concentration during the oestrous cycle, which is not related to the circulating progesterone concentration but could be regulated by oestradiol. We also provide evidence for a diurnal rhythm in the release of vasopressin into the plasma in the ewe.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of active immunization of heifers against inhibin on plasma FSH concentrations, ovarian follicular development and ovulation rate. To study the effects of immunoneutralization of endogenous inhibin on gonadotrophin secretion and ovarian function, prepubertal heifers (n = 6) were actively immunized against a synthetic peptide replica of the N-terminal sequence of bovine inhibin alpha subunit (bI alpha(1-29)Tyr30) coupled to ovalbumin. In contrast to ovalbumin-immunized controls (n = 6), bI alpha(1-29)Tyr30-immunized heifers had detectable inhibin antibody titres (% binding to 125I-labelled bovine inhibin at 1:2000 dilution of plasma) of 17 +/- 3% (S.E.M.) at puberty, rising to 31 +/- 5% by the end of the study period 7 months later. Neither age (immunized: 295 +/- 8 days; controls: 300 +/- 5 days) nor body weight (immunized: 254 +/- 13 kg; controls 251 +/- 9 kg) at onset of puberty differed between groups. Although the difference did not reach statistical significance, mean plasma FSH concentrations recorded in inhibin-immunized heifers remained 35-40% higher than in controls throughout the 12-week period leading up to puberty (P = 0.14) and during nine successive oestrous cycles studied after puberty (P = 0.10). Plasma LH concentrations did not differ between groups at any time during the study. Inhibin immunization had no effect on oestrous cycle length (immunized: 19.8 +/- 0.5 days; controls: 19.9 +/- 0.5 days).(ABSTRACT TRUNCATED AT 250 WORDS)


3 beta-hydroxysteroid dehydrogenase inhibitor reduces ovarian steroid production but increases ovulation rate in the ewe: interactions with gonadotrophins and inhibin. Two experiments were carried out during the breeding season in ewes, first to investigate the effects of oral administration of a 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) inhibitor (epostane) on the number of corpora lutea, and secondly to investigate the mechanism through which epostane acts. In the first experiment Dorset Horn ewes were treated orally with 25, 50, 100 or 200 mg epostane twice daily between days 10 and 15 of the oestrous cycle. All doses of epostane resulted in an increase in the number of corpora lutea per ewe, although the response was curvilinear, with the 25 mg dose showing the largest response and the 200 mg group the smallest response. Although there was no difference between groups in the number of ewes showing oestrus, the higher doses of epostane had a detrimental effect on fertility. In the second experiment Welsh Mountain ewes were treated twice daily with 25 mg epostane from day 10 of the oestrous cycle and the ovaries were removed for analysis during either the luteal or the follicular phases. Treatment significantly increased the number of follicles greater than 6 mm in diameter, but significantly reduced in-vitro follicular oestradiol and testosterone production. Despite a marked increase in peripheral inhibin concentrations there was no effect on in-vitro inhibin production. Epostane treatment also caused a significant reduction in peripheral FSH concentrations and an increase in mean LH concentration. The latter was due to an increase in LH pulse frequency during the luteal phase and LH pulse amplitude during the follicular phase. These results confirm that treatment of ewes with epostane orally has a significant effect on follicular steroidogenesis and causes a significant increase in the number of corpora lutea per ewe. This effect on ovulation rate is not via an increase in peripheral FSH concentration, but may be caused by a reduction in follicular steroid activity either directly on the ovary or via an alteration in the pattern of LH secretion.


Tissue kallikrein is associated with prolactin-secreting cells within human growth hormone-secreting adenomas. Tissue kallikrein is a serine protease which may be involved in the intracellular processing of prolactin in the anterior pituitary gland. The expression of tissue kallikrein, in the rat, is promoted by oestrogen and inhibited by dopamine. Human and rat prolactinomas contain markedly increased amounts of tissue kallikrein; this is comparatively reduced if patients are pretreated with the dopamine agonist, bromocriptine, before surgery. Some GH-secreting adenomas are mixed and also contain prolactin-secreting cells. We therefore investigated 27 GH-immunostaining human pituitary adenomas for the presence of immunoreactive tissue kallikrein. Sixteen of the adenomas had positive immunostaining for prolactin; eight of these patients had associated clinical hyperprolactinaemia before the tumour was removed. Tissue kallikrein immunoreactivity was found in ten adenomas, all of which also had prolactin immunopositivity. There was a close relationship between the percentage of cells staining for prolactin and tissue kallikrein but not for GH. A further eight adenomas had patchy positivity, i.e. less than 1% of cells immunostained for tissue kallikrein and six of these also had some prolactin-staining cells. Nine out of eleven purely GH-staining adenomas had no tissue kallikrein immunopositivity, the remaining two showing patchy staining. A review of bromocriptine responsiveness, as assessed by mean GH hormone levels during oral glucose tolerance tests before and after therapy was commenced, indicated that patients with adenomas which stained for prolactin and tissue kallikrein were more likely to respond to bromocriptine than those which failed to do so.


The supraoptic nucleus in the pig hypothalamus: postnatal development and the effect of gonadal steroids. Postnatal development of the supraoptic nucleus (SON) in the pig hypothalamus was studied morphometrically. The volume of the SON increased from 6.2 +/- 0.45 (S.E.M.) mm3 at 7 weeks postnatally to 18.5 +/- 1.35 mm3 at 2.5 years of age. A sex difference was found at the development point when the SON volume increased, with earlier SON enlargement in males. This sex difference was 30% at 30 weeks and 50% at 1 year of age. At 2.5 years of age no difference in volume was apparent between the sexes. The number of SON neurones was similar for all age groups concerned (43,500 +/- 1475), except for the 2.5-year-old females where 40% more were found (55,500 +/- 3285). No significant difference was found in neurone number between gonadectomized and sham-operated animals, but the operation caused a 30% reduction in the number of neurones and SON volume. Testosterone supplementation following gonadectomy, during the first 4 weeks postnatally, resulted in sexual dimorphism, the males having more SON neurones than the females. The volume showed only a trend in the same direction. Testosterone supplementation at other ages did not result in any difference when compared with controls. The results of this study show that the postnatal development of the SON of the pig is sexually dimorphic, and that it continues after puberty in females. In contrast to the vasopressin- and oxytocin-containing nucleus, the development of the SON was not influenced by gonadectomy and only slightly by gonadal steroids.


Concanavalin-A-bound and -unbound prolactin in normal and hyperprolactinaemic rats. Concanavalin-A (Con-A)-bound and -unbound forms of prolactin were studied in female Wistar-Furth rats, both normal and with hyperprolactinaemia induced by treatment with oestrogen or a prolactinoma graft. In normal rats, Con-A-bound prolactin was the major circulating form (more than 50%) and a minor pituitary component (less than 10%), essentially as 25 kDa prolactin. In oestrogen-treated rats, plasma prolactin levels were 100-fold higher and pituitary weight was fivefold higher than in the controls, but total pituitary prolactin content was unmodified. Under oestrogen, Con-A-bound prolactin represented about one-third of the total hormone levels in the plasma and less than 10% in the pituitary. In the pituitary, bound prolactin was found essentially as 25 kDa and unbound prolactin as 22, 30 and 40-45 kDa. A similar increase in plasma prolactin levels was induced 6 months after the graft of a prolactinoma. Pituitary weights and total pituitary prolactin contents were slightly decreased. Plasma and pituitary Con-A-bound prolactin levels were similar to those observed in oestrogen-treated rats. On the other hand, unbound prolactin was only present as a 22 kDa monomer. In the tumour, Con-A-bound prolactin (essentially as 25 kDa prolactin) represented one-third of the total hormone level and unbound prolactin was composed of the 22 and 45 kDa forms, this latter form being partially transformed into 22 kDa by heating.(ABSTRACT TRUNCATED AT 250 WORDS)


Antiprogesterone RU486 induces dissociation of LH and FSH secretion in the cyclic rat: effect of anti-inhibin serum. Administration of the antiprogesterone RU486 to 4-day cyclic rats from metoestrus to pro-oestrus increases serum levels of LH while decreasing levels of FSH. If it is assumed that there is only one gonadotrophin-releasing hormone, there is no direct explanation for the decrease in FSH concentrations. The purpose of these experiments was to investigate the effect of RU486 on gonadotrophin secretion in cyclic rats during periods when the secretion of LH and FSH diverges. RU486 blunted the transient increase in FSH concentration on the afternoon of metoestrus and the compensatory ovarian hypertrophy on the next day of oestrus in unilaterally ovariectomized 4-day cyclic rats. In addition, bilateral ovariectomy reversed the effect of RU486 on the basal secretion of FSH. RU486 induced an increase in basal LH concentrations. Since ovarian inhibin decreases the basal release of FSH, and decreases in peripheral inhibin seem to be responsible for the transient rise in FSH during the oestrus cycle, the effect of RU486 on serum levels of LH and FSH during dioestrus in rats injected with a sheep anti-inhibin serum (AIS) were further evaluated. Treatment with AIS increased FSH levels in oil-treated rats without altering the levels of LH. In contrast, the effects of AIS on FSH secretion were blunted in RU486-treated rats. The results suggest that inhibin might be involved in the RU486-induced decrease of FSH secretion in cyclic rats.


Antiprogesterone RU486 increases serum immunoreactive inhibin levels and LH:FSH and testosterone:oestradiol ratios in cyclic rats. Since administration of the antiprogesterone RU486 to cyclic rats results in a dissociation of basal LH and FSH secretion we studied its effects on peripheral levels of inhibin, oestradiol and testosterone throughout the oestrous cycle. Cyclic rats were given RU486 (2 mg) twice daily (09.00 and 17.00 h) on metoestrus, dioestrus and pro-oestrus. Oil-treated rats were used as controls. Serum concentrations of immunoreactive inhibin in oil-treated rats increased from metoestrus to pro-oestrus and decreased at oestrus. RU486-treated rats had serum inhibin concentrations significantly increased over oil-treated rats at dioestrus and pro-oestrus, but not at oestrus. At both pro-oestrus and oestrus serum concentrations of LH, testosterone and oestradiol were significantly raised in RU486-treated rats compared with oil-treated controls. In contrast, serum FSH concentrations in RU486-treated rats were decreased on both days. Ovaries from RU486-treated rats showed an increased testosterone content at pro-oestrus, mainly in the interstitial tissue. The results of the present study demonstrate that RU486 has a stimulatory effect on inhibin secretion, and offer an explanation for the decrease in basal serum FSH levels. The low FSH secretion on the morning of oestrus in spite of the low levels of inhibin suggests that progesterone is involved in FSH secretion at this time.


Short-term changes in the 3,5,3'-tri-iodothyronine nuclear receptor-binding capacity of porcine skeletal muscle following food intake. The time-course of changes in the nuclear 3,5,3'-tri-iodothyronine (T3) receptor-binding capacity (Bmax) of longissimus dorsi muscle has been investigated in cold-acclimated young pigs after a single large meal. Measurement of Bmax values 4, 8, 12 and 24 h after feeding indicated a decline in receptor numbers after food intake with the lowest values occurring at 8 h. The receptor numbers then increased significantly, with the values at 12 h being more than 50% higher than those obtained at 8 h. The Bmax values reached their highest level 24 h after feeding. No significant changes in the dissociation constant were observed. Possible reasons for the changes in T3 receptor numbers are discussed and it is suggested that the increase in T3 receptor numbers 12-24 h after feeding may play a role in enhancing the thermogenic capacity of the tissues in response to food.


Discordant secretion of relaxin by individual porcine large luteal cells: quantitative analysis by a reverse haemolytic plaque assay. Individual large luteal cells (LLC) derived from pregnant swine differ conspicuously in their ability to secrete the peptide hormone relaxin under basal and stimulated conditions--the phenomenon of functional heterogeneity. The purpose of this study was to quantitate knowledge of this phenomenon through use of a reverse haemolytic plaque assay, a technique that utilizes antibody-directed, complement-mediated erythrocyte lysis to detect hormone secretion by single LLCs in culture. Measurement of individual plaque areas (an index of the amount of relaxin secreted) demonstrated an approximate 100-fold range in the amount of relaxin secreted by a single cell under basal conditions. This range was doubled by exposure to the phorbol ester, 4 beta-phorbol 12 beta-myristate 13 alpha-acetate (PMA; 50 nmol/l). Under basal conditions, 50 and 80% of the total amount of relaxin was secreted by approximately 10 and 30% of all LLCs respectively. The size of these fractions was not influenced by the time of incubation (1-8 h), or by the presence of either of two non-specific stimulatory relaxin secretagogues, PMA (50 nmol/l) or arachidonic acid (1 mumol/l). The unimodal frequency distribution of plaque areas (under basal or stimulated conditions) suggests that relaxin-secreting LLCs comprise a discrete functional population of secretory cells, at least under these experimental conditions. We conclude that a remarkably small fraction of LLCs secretes the majority of relaxin, and that the size of this fraction was not influenced by time or secretagogues.


The ontogenic and functional relationships between growth hormone- and prolactin-releasing cells during the development of the bovine pituitary. The ontogeny of GH- and prolactin-releasing cells in the developing bovine pituitary was evaluated by reverse haemolytic plaque assays which allows for the detection of hormone release from individual pituitary cells in culture. With this approach, we observed that GH-releasing cells ontogenically preceded prolactin-releasing cells. In fact, GH secretors were observed as early as 59 days of gestational age while cells that released prolactin were not identified until 98 days. The amounts of both GH- and prolactin-releasing cells increased with time to reach more than 50% and 20% of all pituitary cells near term (term approximately 280 days) respectively. Interestingly, the first cells shown to release prolactin also released GH (i.e. were mammosomatotropes). This temporal and functional relationship between GH and prolactin secretors provides suggestive evidence that GH-secreting cells act as the progenitor cells for prolactin secretors via a functional intermediate, the mammosomatotrope.


Effects of gonadotrophin-releasing hormone, recombinant human activin-A and sex steroid hormones upon the follicle-stimulating isohormones secreted by rat anterior pituitary cells in culture. FSH is produced and secreted from the anterior pituitary gland of rats in multiple molecular forms. At times of high gonadotrophin-releasing hormone (GnRH) and oestrogen output (e.g. the morning of the day of pro-oestrus) the pituitary increases the production of FSH isoforms with isoelectric point (pI) values greater than 5.0, whilst sex steroid deprivation leads to the production of strongly acidic and less in-vitro biologically active FSH molecules. It is not known, however, whether sex steroids modulate the production of specific FSH isoforms by a direct action at the pituitary level or indirectly through altering the rate of synthesis and/or secretion of GnRH. In order to obtain some insight on this issue, we examined the charge heterogeneity of FSH secreted by cultured pituitary cells exposed to different FSH-releasing factors, oestradiol-17 beta and progesterone, alone or in different time-sequenced combinations. Anterior pituitary glands from 21-day-old female rats were enzymatically dispersed into a single cell suspension and cultured for 5 days. During days 1 to 3, cells were incubated in the absence of factors or steroid hormones; on days 3 to 4, cells were incubated in the absence (controls) or presence of either oestradiol-17 beta (3.67 nmol/l) or oestradiol-17 beta plus progesterone (3.67 and 31.8 nmol/l respectively). Finally, during days 4 to 6, GnRH (10 nmol/l) or recombinant human activin-A (2 nmol/l) were added to half of all culture wells. Media from each cell group were concentrated and the several forms of secreted FSH were then separated by polyacrylamide gel isoelectric focusing (pH range 6.5-4.0) and quantitated. All media concentrates contained several forms of immunoactive secreted FSH focusing within a pH range of 6.44-4.23. A large amount (51-76%) of total FSH recovered focused within a pI range of 4.9-4.0 (area 3), whilst 20-43% and 4-8% of the total were identified within pI range of 5.9-5.0 (area 2) and 6.5-6.0 (area 1) respectively.(ABSTRACT TRUNCATED AT 400 WORDS)


Desialylation of lysosomal membrane glycoproteins by Trypanosoma cruzi: a role for the surface neuraminidase in facilitating parasite entry into the host cell cytoplasm. Trypanosoma cruzi enters host cells via formation of an acidic vacuole which is subsequently disrupted, allowing the parasite access to the cytoplasm. We show that in an acid environment, release of the parasite surface neuraminidase is enhanced, and this release is likely mediated by a phosphatidylinositol-specific phospholipase C (PIPLC), since antibodies to a carbohydrate epitope (CRD) revealed in glycosylphosphatidylinositol (GPI)-anchored proteins after PIPLC cleavage remove the great majority of the soluble neuraminidase activity from culture supernatants. The neuraminidase is active at acidic pH, and is capable of desialylating known vacuolar constituents, i.e., lysosomal membrane glycoproteins. Parasite escape into the cytoplasm is significantly facilitated in terminal sialylation-defective mutant Lec 2 cells, and enzymatically desialylated membranes are more susceptible to lysis by a parasite hemolysin previously implicated in vacuole membrane rupture. These findings provide evidence that terminal sialylation on carbohydrate moieties contributes to maintaining lysosomal membrane integrity, and indicate a role for a protozoan-derived neuraminidase in facilitating parasite entry into host cells. These observations raise the possibility that other microbial neuraminidases may serve a similar function in acidic intracellular compartments.


Switch recombination breakpoints are strictly correlated with DNA recognition motifs for immunoglobulin S gamma 3 DNA-binding proteins. The deletion looping out model of switch (S) recombination predicts that the intervening DNA between switch regions will be excised as a circle. Circular excision products of immunoglobulin switch recombination have been recently isolated from lipopolysaccharide (LPS)-stimulated spleen cells. The recombination breakpoints in these large circles were found to fall within switch regions. Since switch recombination is clearly focused on switch regions, we hypothesized that some DNA-binding protein factor might be involved in specifically recognizing and facilitating the alignment of switch regions before recombination. Two DNA-binding proteins that specifically interact with two discrete regions of the S gamma 3 tandem repeat have been identified in crude and partially purified nuclear extracts derived from LPS- and dextran sulfate (DxS)-activated splenic B cells. The first factor has been found indistinguishable from NF-kappa B by mobility shift assays, methylation interference, competition binding studies, and supershift analysis using an antiserum specific for the p50 component. The second appears to be composed of two closely traveling mobilities that do not separate upon partial purification. This second complex is unique and specific for S gamma 3 by methylation interference assays and competition-binding analysis. The sites at which recombination occurs in the S gamma 3 switch region have been analyzed and found to strictly correlate with the binding sites of the S gamma 3 switch binding proteins.


Association of the tyrosine kinase LCK with phospholipase C-gamma 1 after stimulation of the T cell antigen receptor. Stimulation of the T cell antigen receptor (TCR) activates a protein tyrosine kinase and leads to the tyrosine phosphorylation of phosphoinositide-specific phospholipase C-gamma 1 (PLC gamma 1). The molecular interactions involved in this phosphorylation are not known. After stimulation of the TCR on Jurkat T cells, tyrosine-phosphorylated proteins of 36, 38, 58, and 63 kD coprecipitate with PLC gamma 1. An identical pattern of proteins precipitate with TrpE fusion proteins that contain the Src homology (SH) 2 domains of PLC gamma 1, indicating that these regions of PLC gamma 1 are responsible for binding. TCR stimulation leads to an association between the SH2 domains of PLC gamma 1 and a protein tyrosine kinase, which, by peptide mapping, is identical to p56lck. These studies establish that p56lck associates with PLC gamma 1 as a result of TCR stimulation of Jurkat cells, suggesting that p56lck plays a central role in coupling the TCR to the activation of PLC gamma 1.


Unexpected effects of the severe combined immunodeficiency mutation on murine lymphomagenesis. Strain C.B17 scid/scid (SCID) mice, which lack functional T and B lymphocytes, show heightened susceptibility to the induction of thymic lymphomas by x-irradiation. Susceptibility is highest in thymus-chimeric SCID-BL mice (thymectomized SCID mice bearing a C57BL thymus graft). All SCID-BL lymphomas originate in the cells of the thymic graft (C57BL type) and lack murine leukemia virus expression. Both SCID and SCID-BL lymphomas are phenotypically CD4-8+ and/or CD4+8+, but only the SCID-BL tumors express CD3. Injection of C57BL or BALB/c bone marrow into irradiated SCID-BL mice prevents lymphoma development, but SCID marrow is completely ineffective. The results suggest that the scid condition enhances the activity of a putative lymphomagenic agent induced in the bone marrow by x-irradiation and that C57BL thymic cells are highly sensitive targets. Moreover, the failure of SCID bone marrow to protect against lymphomagenesis vs. the efficacy of marrow from immunocompetent donors points to involvement of T or B lineage cells in this process.


Correlation between lymphocyte-induced donor-specific tolerance and donor cell recirculation. Intravenous infusion of mice with major histocompatibility complex (MHC) incompatible lymphocytes can inhibit the response of recipient T cells capable of recognizing the injected cells, and can enhance survival of grafts sharing MHC with the injected cells. However, neither T cell inactivation nor graft survival enhancement is always achieved. This is particularly true for donor cells that are fully allogeneic (as compared to semiallogeneic) to the recipient. We show here that both donor-specific induced response reduction and graft survival enhancement are directly correlated with the ability of the injected lymphoid cells to persist in the recirculating lymphocyte pool of the host. Whether donor cells persist correlates inversely with the level of natural killer cell (NK) activity in the host. Fully allogeneic cells can only persist in hosts with low NK activity and can then induce response reduction. Both persistence and response reduction are abrogated by injection of the host with poly-I:C, a treatment that boosts host NK activity. The same treatment also destroys the ability of semiallogeneic injected cells to persist, to induce response reduction, and to enhance skin graft survival.


A major surface protein on group A streptococci is a glyceraldehyde-3-phosphate-dehydrogenase with multiple binding activity. The surface of streptococci presents an array of different proteins, each designed to perform a specific function. In an attempt to understand the early events in group A streptococci infection, we have identified and purified a major surface protein from group A type 6 streptococci that has both an enzymatic activity and a binding capacity for a variety of proteins. Mass spectrometric analysis of the purified molecule revealed a monomer of 35.8 kD. Molecular sieve chromatography and sodium dodecyl sulfate (SDS)-gel electrophoresis suggest that the native conformation of the protein is likely to be a tetramer of 156 kD. NH2-terminal amino acid sequence analysis revealed 83% homology in the first 18 residues and about 56% in the first 39 residues with glyceraldehyde-3-phosphate dehydrogenase (GAPDH) of eukaryotic or bacterial origin. This streptococcal surface GAPDH (SDH) exhibits a dose-dependent dehydrogenase activity on glyceraldehyde-3-phosphate in the presence of beta-nicotinamide adenine dinucleotide both in its pure form and on the streptococcal surface. Its sensitivity to trypsin on whole organism and its inability to be removed with 2 M NaCl or 2% SDS support its surface location and tight attachment to the streptococcal cell. Affinity-purified antibodies to SDH detected the presence of this protein on the surface of all M serotypes of group A streptococcal tested. Purified SDH was found to bind to fibronectin, lysozyme, as well as the cytoskeletal proteins myosin and actin. The binding activity to myosin was found to be localized to the globular heavy meromyosin domain. SDH did not bind to streptococcal M protein, tropomyosin, or the coiled-coil domain of myosin. The multiple binding capacity of the SDH in conjunction with its GAPDH activity may play a role in the colonization, internalization, and the subsequent proliferation of group A streptococci.


T cell receptor-mediated recognition of self-ligand induces signaling in immature thymocytes before negative selection. Shaping of the T cell repertoire by selection during intrathymic maturation involves T cell receptor (TCR) recognition of major histocompatibility complex/self-antigen complexes. In this communication, we studied the ability of minor lymphocyte stimulating (Mls) determinants to act as self-tolerogens in the selection of the T cell repertoire. We demonstrate that unprimed T cells from normal as well as TCR transgenic mice form Mls-specific conjugates with antigen-presenting cells, and that this TCR-ligand interaction leads to elevation of intercellular Ca2+ ([Ca2+]i). Peripheral T cells from TCR transgenic mice expressing receptors specific for self-Mls antigen show no reactivities to Mlsa. However, a proportion of immature thymocytes from these mice show specific binding and strong [Ca2+]i elevation in response to self-antigen-presenting cells, although these thymocytes do not proliferate. This self-reactivity of thymocytes is inhibited by antibodies specific for TCR, CD4, CD8, class II molecules, lymphocyte function-associated antigen 1, and intercellular adhesion molecule 1. These results demonstrate for the first time that before thymic negative selection, immature T cells can specifically interact with cells bearing self-antigen, and suggest that the resulting TCR-dependent signal transduction events provide a basis for negative selection of self-reactive T cells.


Natural occurrence and origin of somatically mutated memory B cells in mice. While most murine peripheral B cells express germline-encoded antibodies of classes M and D (mu+ delta+ cells), small numbers of memory B cells expressing somatically mutated immunoglobulin G antibodies are generated upon T cell-dependent immunization. Analyzing the antibody repertoire of the mu-delta- B cell pool in unimmunized mice, we show that these cells express somatically mutated VH genes and that most of these genes derive from a set of germline VH genes dominantly expressed by mu+delta+ B cells. Thus, class-switched memory B cells are generated in the absence of intentional immunization, presumably in response to environmental antigens. These cells are either recruited from mu+delta+ B cells or selected from newly arising B cells in parallel to the latter, by the same antigens.


Two subsets of epithelial cells in the thymic medulla. Information was sought on the features of epithelial cells in the murine thymic medulla. The expression of major histocompatibility complex (MHC) molecules on medullary epithelium was defined by light microscopy with the aid of bone marrow chimeras and MHC-transgenic mice. A proportion of medullary epithelial cells was found to show conspicuously high expression of conventional MHC (H-2) class I (K, D, L) and class II (I-A, I-E) molecules. These cells express a high density of the Y-Ae epitope, a complex of an E alpha peptide and I-Ab molecules found on typical bone marrow-derived cells. MHC+ medullary epithelial cells show limited expression of I-O molecules, a class of atypical nonpolymorphic MHC-encoded class II molecules present on B cells. Other medullary epithelial cells express a high density of I-O molecules but show little or no expression of typical MHC class I or II molecules. MHC and I-O expression thus appear to subdivide medullary epithelial cells into two phenotypically distinct subsets. This applies in adults. In the embryonic thymus most medullary epithelial cells express both types of molecules.


The human immunodeficiency virus gp120 binding site on CD4: delineation by quantitative equilibrium and kinetic binding studies of mutants in conjunction with a high-resolution CD4 atomic structure. The first immunoglobulin V-like domain of CD4 contains the binding site for human immunodeficiency virus gp120. Guided by the atomic structure of a two-domain CD4 fragment, we have examined gp120 interaction with informative CD4 mutants, both by equilibrium and kinetic analysis. The binding site on CD4 appears to be a surface region of about 900 A2 on the C" edge of the domain. It contains an exposed hydrophobic residue, Phe43, on the C" strand and four positively charged residues, Lys29, Lys35, Lys46, and Arg59, on the C, C', C", and D strands, respectively. Replacement of Phe43 with Ala or Ile reduces affinity for gp120 by more than 500-fold; Tyr, Trp, and Leu substitutions have smaller effects. The four positively charged side chains each make significant contributions (7-50-fold). This CD4 site may dock into a conserved hydrophobic pocket bordered by several negatively charged residues in gp120. Class II major histocompatibility complex binding includes the same region on CD4; this overlap needs to be considered in the design of inhibitors of the CD4-gp120 interaction.


Characterization of the interleukin 2 receptors (IL-2R) expressed on human natural killer cells activated in vivo by IL-2: association of the p64 IL-2R gamma chain with the IL-2R beta chain in functional intermediate-affinity IL-2R. Interleukin 2 (IL-2) receptors expressed on the surface of activated T cells and natural killer (NK) cells exhibit a variety of affinity states depending on their subunit composition. Low-affinity binding is associated with a 55-kDa alpha chain, intermediate-affinity binding with a 70-75-kD beta chain, and high-affinity binding with a bimolecular complex of the alpha and beta subunits. In a previous study of the IL-2 receptors expressed on NK cells obtained from cancer patients after in vivo IL-2 therapy, we documented a discrepancy between the level of beta chain and the level of intermediate-affinity IL-2 binding sites expressed on the cell surface. Based on this result, we postulated that formation of intermediate-affinity receptor sites required a component in addition to the beta chain, and that this component was present at limiting levels in the patient NK cells. In the present study we have examined the structure of the intermediate-affinity receptor complex using monoclonal antibodies that recognize the beta chain, but that do not interfere with its ability to bind IL-2. Evidence is presented establishing the physical association of a novel protein of 64 kD with the beta chain in intermediate-affinity IL-2 binding sites. This molecule, termed IL-2R gamma chain, coprecipitated with beta chains prepared from cells that had been incubated with IL-2, but was undetectable in immunoprecipitates prepared in the absence of IL-2. Examination of gamma chain expression in post-IL-2 therapy NK cells, where only low levels of intermediate-affinity IL-2 binding were detectable, revealed that the gamma chain was associated with, on average, only 10-12% of the beta chains expressed on such cells. This contrasted with approximately equal levels of beta and gamma chain expression on YT cells, a cell line that has both high levels of cell surface beta chain expression and high levels of IL-2 binding. Thus, the ratio of gamma chain to beta chain present in the immunoprecipitates roughly correlated with the proportion of beta chain involved in intermediate-affinity receptor sites. This result suggests that the 64-kD gamma chain is the component responsible for regulating the affinity of IL-2 association with the beta subunit. By further defining the structural components necessary for IL-2 receptor formation, these studies provide additional insight into mechanisms whereby lymphocytes might regulate their responsiveness to IL-2.


Systemic lupus erythematosus patients with central nervous system involvement show autoantibodies to a 50-kD neuronal membrane protein. An antibody was detected in the sera from patients with systemic lupus erythematosus (SLE) and central nervous system (CNS) involvement that reacted with a 50-kD antigen in the plasma membrane of brain synaptic terminals. The 50-kD antigen was solubilized with Triton X-100 from preparations enriched with synaptic plasma membranes, and was partially purified by molecular sieve filtration column chromatography. The sera of 19 of 20 CNS-SLE patients showed strong to moderate immunoreactivity with the 50-kD protein in Western blots. Immunoreactivity with the 50-kD protein was also detected in the cerebrospinal fluid of CNS-SLE patients. Control sera from healthy individuals did not react with the 50-kD protein. Low to background reactivity was detected in 35% of a group of SLE patients without CNS manifestations, and in 3% of patients displaying other connective tissue diseases. A total of 100 individuals were tested in this study. Purified autoantibodies to the 50-kD protein from CNS-SLE patients were used for immunofluorescent labeling of neuroblastoma cells. The immunofluorescent staining revealed a distinct macular distribution pattern on the surface of the cell membrane. Taken together, the data suggest that the 50-kD protein may be an important target for autoantibodies, preponderantly found in CNS-SLE patients, and that the antigen may play a role in the pathogenesis of some neurological manifestations in SLE.


Memory T cells are anergic to the superantigen staphylococcal enterotoxin B. We have used staphylococcal enterotoxin B (SEB) to study the role of naive and memory T cells in the induction of peripheral tolerance. After administration of SEB to mice, the numbers of naive and memory T cells increase, as does the proportion of memory T cells, which are unresponsive to further stimulation with SEB in vitro. In addition, memory T cells generated in response to conventional antigen, which proliferate and provide help to B cells in the presence of the conventional antigen, fail to respond to superantigen. Hence, memory T cells, in general, are anergized by SEB. These results suggest that SEB-induced activation and anergy reflect the combined responses of naive and memory T cells. The differential activation vs. anergy of naive and memory T cells by superantigen may be related to cytokine production and may play an important role in the etiology of autoimmune diseases or immunodeficiency diseases such as acquired immune deficiency syndrome.


Growth of epithelial cells in the thymic medulla is under the control of mature T cells. Epithelial cells in the thymic medulla are conspicuous in normal adult mice, but sparse in the early fetal thymus and the thymus of adult T cell-deficient SCID mice. To examine whether growth of medullary epithelial cells (MEC) depends upon local contact with mature T cells, we used the finding that the SCID thymus is unusually permeable to mature T cells entering from the bloodstream. When SCID mice received multiple injections of mature lymph node T cells from birth, the thymus accumulated large numbers of mature TCR+ T cells of resting phenotype, but contained virtually no immature (CD4+8+) cells. The injected T cells localized in the medullary region of the thymus and led to marked regeneration of MEC. These and other data suggest that the growth of MEC is under the control of mature T cells. Placing MEC under T cell control might be a device for regulating the size and integrity of the medulla, especially during the phase of rapid thymic growth. Maintaining the cellular components of the medulla in proper balance could be critical for ensuring efficient self tolerance induction.


Inhibition of tumor growth in vivo with a soluble CD44-immunoglobulin fusion protein. CD44H is the principal cell surface receptor for hyaluronate, which is a major glycosaminoglycan of the extracellular matrix. Expression of CD44H is enhanced in a variety of malignant tumors and correlates with tumor aggressiveness, supporting the notion that interaction between CD44H and hyaluronate may play an important role in tumor growth and dissemination. In this report we show that in vivo tumor formation by human lymphoma Namalwa cells, stably transfected with CD44H, can be suppressed by a soluble human CD44H-immunoglobulin fusion protein. Disruption of the interaction between CD44H and its physiologic ligands may provide a novel strategy for controlling tumor growth in vivo.


Evaluation of the functional equivalence of major histocompatibility complex class II A and E complexes. Most mice display two conventional major histocompatibility complex class II isotypes, A and E. Several A+E- strains have been observed, but never any that are A-E+. Because of this and because of hints from several lines of functional analysis, it has been proposed that the two isotypes might not operate equivalently. This proposition has not been directly testable until now because of the lack of an E-only strain. We report the production of such mice, exploiting previously created class II-transgenic and class II-"knock-out" lines. A+E-, A-E-, and A-E+ littermates have been compared by a number of parameters. We find that E and A molecules are, for the most part, functionally equivalent. However, subtle differences are seen in their ability to engage CD4 molecules on immature thymocytes, and in the profile of receptors on T cells selected into the periphery.


Biological movements look uniform: evidence of motor-perceptual interactions. Six experiments demonstrate a visual dynamic illusion. Previous work has shown that in 2-dimensional (2D) drawing movements, tangential velocity and radius of curvature covary in a constrained manner. The velocity of point stimuli is perceived as uniform if and only if this biological constraint is satisfied. The illusion is conspicuous: The variations of velocity in the stimuli exceed 200%. Yet movements are perceived as uniform. Conversely, 2D stimuli moving at constant velocity are perceived as strongly nonuniform. The illusion is robust: Exposure to true constant velocity fails to suppress it. Results cannot be explained entirely by the kinetic depth effect. The illusion is evidence of a coupling between motor and perceptual processes: Even in the absence of any intention to perform a movement, certain properties of the motor system implicitly influence perceptual interpretation of the visual stimulus.


The role of balance dynamics in the active perception of orientation. The Stoffregen and Riccio (1988) hypothesis that perceived orientation is determined primarily by balance dynamics was tested. Perception of orientation was evaluated in the context of a task that required Ss to control the roll orientation of a device in which they were seated. The device's direction of balance was manipulated across trials and thus was independent of gravity. Eighteen Ss participated in the investigation. After each trial, Ss estimated their mean tilt with respect to upright. Correlations of perceived tilt with tilt from balance were consistently higher than the correlations with gravity tilt. The dominance of balance over gravity depended on the magnitude of tilt from balance.


Symmetry breaking dynamics of human multilimb coordination. The dynamics of pattern formation and change are studied in a complex multicomponent system, specifically the arms and legs of human Ss. Among the novel features observed are differential stability of coordinative modes produced by limbs moving in the same versus different directions (Experiment 1); transitions between coordinative modes preceded by a slow drift in relative phase (Experiments 1 and 2); bifurcations or phase transitions from 1 four-limb pattern to another (Experiment 2); and spontaneous emergence of non-1:1-frequency- and phase-locked patterns, in addition to periods of relative coordination (Experiment 3). All observed relative phasing patterns and their dynamics (stability, loss of stability, intermittency) are shown to arise from the same underlying nonlinear dynamical structure, an important feature of which is broken symmetry.


Influence of animation on dynamical judgments. The motions of objects in the environment reflect underlying dynamical constraints and regularities. The conditions under which people are sensitive to natural dynamics are considered. In particular, the article considers what determines whether observers can distinguish canonical and anomalous dynamics when viewing ongoing events. The extent to which such perceptual appreciations are integrated with and influence common-sense reasoning about mechanical events is examined. It is concluded that animation evokes accurate dynamical intuitions when there is only 1 dimension of information that is of dynamical relevance. This advantage is lost when the observed motion reflects higher dimension dynamics or when the kinematic information is removed or degraded.


Changing affordances in stair climbing: the perception of maximum climbability in young and older adults. This experiment extended Warren's leg-length model by investigating the relevance of leg strength and joint flexibility on perceptual judgments of climbability. From a set of 8 stairs (riser heights: 38-91 cm), 24 older and 24 young adults were asked to identify the highest stair they could climb without using their hands or knees. Ss then attempted to climb the selected stair. Tall and short young observers perceived similar action boundaries despite leg-length differences. Tall and short older adults had divergent action boundaries when a single-scale leg-length model was applied. A regression model that used flexibility and leg-strength measurements provided a better fit of the older adult data, indicating that models applying functional (kinematic and kinetic) criteria might be useful in describing lawful relationships between organisms and the environment.


Nonvisual judgment of the crossability of path gaps. The ability of persons to locomote without vision with the aid of a hand-held probe was investigated in 4 experiments focusing on the negotiation of gaps in a pathway. In Experiment 1, Ss of extreme heights judged under sighted and blindfolded conditions whether gaps of different sizes could be crossed by walking. Crossability boundaries were lower for shorter Ss, but the height effect disappeared under normalization by leg length. Experiments 2-4 altered mechanical aspects of the probe (length and mass distribution) in the nonvisual case. Ss' performance was still reliable in each experiment, and their performances were constrained by parameters of the gap and of their action systems. Probe variables, however, had surprisingly little influence on the judgments of crossability. Results were discussed in terms of potential roles for information in different perceptual modalities.


Role of the inertia tensor in perceiving object orientation by dynamic touch. Ss wielded an occluded L-shaped rod and attempted to perceive the direction in which the rod was pointing with respect to the hand. The pattern of the rod's different resistances to rotation in different directions, quantified by the inertia tensor, changes systematically with the rod's orientation. Perception of orientation by wielding is possible if the tissue deformation consequences of the rod's inertia tensor are detectable. It was shown that perceived orientation was a linear function of actual orientation for both free and restricted wielding and for rods of different-size branches. The eigenvectors of the inertia tensor were implicated as the basis for this haptic perceptual capability. Results were discussed in reference to information-perception specificity and its implications for effortful or dynamic touch.


The role of spectral and dynamic cues in imagery for musical timbre. The musical quality of timbre is based on both spectral and dynamic acoustic cues. Four 2-part experiments examined whether these properties are represented in the mental image of a musical timbre. Experiment 1 established that imagery occurs for timbre variations within a single musical instrument, using plucked and bowed tones from a cello. Experiments 2 and 3 used synthetic stimuli that varied in either spectral or dynamic properties only, to investigate imagery with strict acoustic control over the stimuli. Experiment 4 explored whether the dimension of loudness is stored in an auditory image. Spectral properties appear to play a much larger role than dynamic properties in imagery for musical timbre.


Perceptual interactions between musical pitch and timbre. These experiments examined perceptual interactions between musical pitch and timbre. Experiment 1, through the use of the Garner classification tasks, found that pitch and timbre of isolated tones interact. Classification times showed interference from uncorrelated variation in the irrelevant attribute and facilitation from correlated variation; the effects were symmetrical. Experiments 2 and 3 examined how musical pitch and timbre function in longer sequences. In recognition memory tasks, a target tone always appeared in a fixed position in the sequences, and listeners were instructed to attend to either its pitch or its timbre. For successive tones, no interactions between timbre and pitch were found. That is, changing the pitches of context tones did not affect timbre recognition, and vice versa. The tendency to perceive pitch in relation to other context pitches was strong and unaffected by whether timbre was constant or varying. In contrast, the relative perception of timbre was weak and was found only when pitch was constant. These results suggest that timbre is perceived more in absolute than in relative terms. Perceptual implications for creating patterns in music with timbre variations are discussed.


Musical duplex perception: perception of figurally good chords with subliminal distinguishing tones. In a variant of duplex perception with speech, phoneme perception is maintained when distinguishing components are presented below intensities required for separate detection, forming the basis for the claim that a phonetic module takes precedence over nonspeech processing. This finding is replicated with music chords (C major and minor) created by mixing a piano fifth with a sinusoidal distinguishing tone (E or E flat). Individual threshold intensities for detecting E or E flat in the context of the fixed piano tones are established. Chord discrimination thresholds defined by distinguishing tone intensity were determined. Experiment 2 verified masked detection thresholds and subliminal chord identification for experienced musicians. Accurate chord perception was maintained at distinguishing tone intensities nearly 20 dB below the threshold for separate detection. Speech and music findings are argued to demonstrate general perceptual principles.


The sum of the parts does not equal the whole: evidence from bihemispheric processing. The 5 experiments in this article examine whether bihemispheric processing can be predicted from the processing observed when information is directed initially to just 1 hemisphere. Ss decided if laterally presented words rhymed with a previously presented central target. Trials varied in the degree to which the information displayed was redundant. On some trials, the same word appeared twice; in others, different words appeared, but they both led to the same decision; and finally, on some trials, different words appeared, each of which also led to a different decision. The patterns found for unilateral and bilateral trials were distinct. Furthermore, the pattern observed on bilateral trials could not be predicted from that found on unilateral trials, suggesting that interhemispheric interaction cannot be surmised from observing a hemisphere processing information in relative isolation.


Global shape cannot be attended without object identification. Whether the global shape of objects can be processed without accessing semantic or identity information was tested. Ss judged which of 2 fragmented forms had the same global shape as a reference stimulus. Matching stimuli could be physically identical, semantically related, or unrelated. The reference stimulus and nonmatching (distractor) form could be semantically related or unrelated. Similarity effects in the related condition were unconfounded with matches nameable and nonnameable forms. For nameable forms, related matching forms facilitated performance; a related distractor disrupted performance. Semantic interference was eliminated when nameable distractors were replaced with nonnameable partners; semantic similarity effects on matching were eliminated with a nonnameable reference stimulus and with inverted targets and distractors. Access to information concerning global shape does not normally occur without object identification.


Reference frame and effects of orientation on finding the tops of rotated objects. Effects of stimulus orientation across trial blocks and the spatial reference frame were investigated with a task in which Ss, with their head upright or tilted, judged a dot to be near the top or the bottom of rotated line drawings of objects. Objects used in this task were also named. Response times from the first block of trials increased linearly for objects rotated from 0 degrees to 120 degrees from the upright. Across blocks, orientation effects diminished for naming but remained the same for top-bottom discriminations. Practice with top-bottom discriminations diminished orientation effects when the same objects were subsequently named. The spatial reference frame for top-bottom discriminations was midway between retinal and environmental coordinates. Specifying the location of object features is of greater importance for top-bottom discriminations than for naming and underlies orientation effects in these tasks.


Selective attention and visual search: revision of an allocation model and application to age differences. The present experiments examined a revised version of the Eriksen and Yeh model of attentional allocation during visual search. The results confirmed the assumption of the model that performance represents a weighted combination of focused- and distributed-attention trials, although Ss relied on focused attention more than was predicted. Consistent with the model, predictions on the basis of the assumption of a terminating search fit the data better than predictions on the basis of an exhaustive search. The effects of varying cue validity favored an interpretation of focused attention in terms of a processing gradient rather than a zoom lens. Although the allocation of attention across trials was similar for young and older adults, there was an age-related increase in the time required to allocate attention within individual trials.


Splitting focal attention.

Splitting focal attention. The study was based on the inverse relationship between the effect of attention on reaction time (RT) and the size of the area over which focal attention is allocated. Independent occurrence of this in 2 locations in the opposite hemifields would be evidence of attention splitting. In Experiment 1, in which the 2 locations were denoted by empty boxes, there was an inverse relationship between size of the stimulated box and RT. Experiment 2 replicated the finding with different stimulation conditions. In Experiment 3, no relationship was found between RT and length of a cuing line. In Experiment 4, in which attention was manipulated by central cues, there was an effect of box size on valid and neutral trials but not invalid trials. Observers could split focal attention and manipulate simultaneously 2 independent attentional foci on objects located in the opposite hemifields.


Temporary suppression of visual processing in an RSVP task: an attentional blink? . Through rapid serial visual presentation (RSVP), we asked Ss to identify a partially specified letter (target) and then to detect the presence or absence of a fully specified letter (probe). Whereas targets are accurately identified, probes are poorly detected when they are presented during a 270-ms interval beginning 180 ms after the target. Probes presented immediately after the target or later in the RSVP stream are accurately detected. This temporary reduction in probe detection was not found in conditions in which a brief blank interval followed the target or Ss were not required to identify the target. The data suggest that the presentation of stimuli after the target but before target-identification processes are complete produces interference at a letter-recognition stage. This interference may cause the temporary suppression of visual attention mechanisms observed in the present study.


Response time distributions and the Stroop Task: a test of the Cohen, Dunbar, and McClelland (1990) model. Cohen, Dunbar, and McClelland's (1990) model was tested for Strooplike interference tasks by studying the shape of the distribution of response latencies produced by Ss and by the model. The model correctly anticipates changes in mean response latency (M(RT)) across congruent and incongruent conditions. It does not, however, correctly anticipate changes in the shape of the distributions, even though changes in the shape of the distributions underlie the changes in M(RT). Thus the model predicts M(RT) successfully but for the wrong reason. It is concluded that the model is not an adequate account of Ss' performance in the Stroop task.


Mechanisms of ion and acid-base regulation at the gills of freshwater fish. This review examines the branchial mechanisms utilized by freshwater fish to regulate internal acid-base status and presents a model to explain the underlying basis of the compensatory processes. Rainbow trout, Oncorhynchus mykiss, and brown bullhead, Ictalurus nebulosus, were examined under a variety of experimental treatments which induced respiratory and metabolic acid-base disturbances. Acid-base regulation was achieved by appropriate adjustments of Na+ and Cl- net fluxes across the gills which, in turn, were accomplished by variable contributions of three different branchial mechanisms: 1) differential changes in Na+ and Cl- diffusive effluxes, 2) changes in internal substrate (H+, HCO3-) availability, and 3) morphological adjustments to the gill epithelium. Differential diffusive efflux of Na+ over Cl- was involved only during periods of metabolic alkalosis. The importance of internal substrate availability was demonstrated using a two-substrate model. According to the model, ionic flux rates (J(in)Cl-, J(in)Na+) are determined not only by the concentration of the external ion (Na+, Cl-) but also by the concentration of the internal counterion (H+, HCO3-). This system provides for an "automatic negative feedback" to aid in the compensation of metabolic acid-base disturbances. Morphological alteration of the gill epithelia and the associated regulation of chloride cell (CC) fractional area is an essential third mechanism which is especially important during respiratory acid-base disturbances. Specifically, fish vary the availability of the CC associated Cl-/HCO3- exchange mechanism by physical covering/uncovering of CCs by adjacent pavement cells.


Intracellular pH regulation and metabolic interactions in hepatic tissues. Intracellular pH (pHi) regulation in the vertebrate liver relies heavily on ionic transport mechanisms. Liver, in common with many tissues, has plasma membrane Na(+)-H+ and Cl(-)-HCO3- electroneutral exchangers which work in opposition to tightly control pHi. Mammalian livers also possess electrogenic Na(+)-HCO3- exchangers, capable of base uptake, which, when coupled to pHi-mediated changes in membrane potential, probably confer an additional measure of pHi control, compared to fish livers, where the transporter appears to be functionally absent. It is suggested that this may be a fundamental difference between aquatic and aerial breathing. pHi regulation has barely been examined in invertebrate hepatic tissues, but already some interesting differences are apparent. Notably, an electrogenic 2Na(+)-1H+ acid-extrusion system is present in apical membranes of crustacean hepatopancreas. Despite these ionic control systems, complex acid-base disturbances (e.g., "metabolic" acidosis) have been known for some time to influence hepatic metabolism in vertebrates, but few studies have carefully examined the independent effects of the acid-base variables involved. Thus mechanistic explanations for the effects of acid-base disturbances are scarce. Ureogenesis in mammals has been well studied, and several pH-related mechanisms are evident. In contrast, the pH-insensitivity of ureogenesis in fish liver may represent a second difference between aquatic and terrestrial species. In summary, by virtue of its metabolic diversity, liver represents a potentially important organ in acid-base balance, and an interesting study tissue for interrelationships between metabolism and acid-base balance.


Cytoskeletal sheets appear as universal components of mammalian eggs. The eggs of two mammalian species have been shown to contain novel cytoskeletal elements, referred to as cytoskeletal sheets, which undergo stage-specific changes in spatial organization at three key developmental transitions, fertilization, compaction, and blastocyst formation. If cytoskeletal sheets have an integral role in these developmental transitions, the sheets should be present in the eggs of other mammals as well. We examined the eggs of four additional species to determine if sheets were present. Our results indicate that sheets were present and they can be categorized into two classes based on their surface appearance. Cytoskeletal sheets in eggs of hamsters and rats have a smooth surface appearance, while eggs from humans, cows, pigs, and mice have a fibrous surface appearance. In addition, we observed that species-specific variations exist in the width of the sheets and in the density of the sheets (i.e., number per micron 2) in the eggs. These species-specific variations may relate to the role of the sheets during early development.