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Y Yabu

Publications and source records attributed to Y Yabu.

At least 37 records · Page 2Linked to original sources

Immune mechanism facilitating clearance of Trypanosoma gambiense by IgG3 antibody from infected host.

The aggregation of trypansomes, immune adherence to macrophage and protection against infection of Trypanosoma gambiense are associated with IgG3 antibody. IgG3-mediated clumping trypanosomes are readily dissociated by the aid of complement to become free. Dissociation of the clumped trypanosomes in the equivalence area released approximately fifty percent of previous bound surface antigens. These antigens were capable of binding again to new IgG3 antibody. Experiments indicated that complement deposition altered functionally bivalent IgG3 antibody in the immune complex into a univalent one. Such event in the presence of complement is of great advantage to the infected host in killing pathogens in vivo, as it allows more antibodies to attach to surface antigens and subsequently initiate complement activity.

Animals↗

An effective axenic culture system for Trypanosoma brucei rhodesiense blood stream forms in vitro.

An effective axenic culture system for Trypanosoma brucei rhodesiense (ILRAD 1501) bloodstream forms is demonstrated. Bloodstream forms were continuously grown in 25 mM HEPES-buffered D-MEM supplemented with 10 microM bathocuproine sulfonate (BCS), 100 microM cysteine, and 20% heat-inactivated fetal bovine serum at 37 degrees C in vitro. At the initiation of the culture, T. b. rhodesiense bloodstream forms required the presence of 0.2 IU/ml insulin and 1 mM pyruvate, while bloodstream forms were grown in the culture medium without these supplements 4 days after initiation of the culture. Under this culture condition, T. b. rhodesiense bloodstream forms increased in number to 7 to 8 x 10(6) trypanosomes/ml, by day 4 after initiation of the culture. The trypanosomes cultured in this axenic system for 150 days were typically long and slender and retained their virulence for mice. This axenic culture system is extremely useful for in vitro cloning of T. b. rhodesiense bloodstream forms in vitro.

Animals↗

Long-term culture and cloning system for Trypanosoma brucei gambiense bloodstream forms in semi-defined medium in vitro.

A new semi-defined medium extremely useful for the long-term cultivation and cloning of Trypanosoma b. gambiense (Wellcome strain) bloodstream forms is described. Bloodstream forms could be continuously grown in 25 mM HEPES-buffered D-MEM supplemented with 10 microM bathocuproine sulfonate (BCS), 100 microM cysteine, and 20% heat-inactivated fetal calf serum at 37 degrees C in vitro. Under these culture conditions, T. b. gambiense bloodstream forms increased in number up to 2-3 x 10(6) trypanosomes/ml by day 3 after initiation of the culture. The trypanosomes maintained in this culture system for 200 days retained their infectivity for mice. Morphologically, they were long and slender, and a surface coat was evident on the cell surface and flagellar membrane. In vitro cloning with single bloodstream forms of T. b. gambiense could be achieved with high efficiency.

Animals↗

Trypsin-stimulated transformation of Trypanosoma brucei gambiense bloodstream forms to procyclic forms in vitro.

The effect of trypsin treatment on the transformation of monomorphic Trypanosoma b. gambiense (Wellcome strain) bloodstream forms to procyclic forms was studied in HEPES-buffered RPMI 1640 medium supplemented with 20% inactivated fetal calf serum in the presence of GA-1 cells as feeder layers at 27 degrees C. In this system, 35%-40% of the bloodstream forms transformed to procyclic forms within 24 h, and over 95% of the trypanosomes changed into procyclic forms by day 3 after initiation of the culture. Established cultures of procyclic forms yielded up to 1.5-2 x 10(7) trypanosomes/ml. However, transformation of nontreated and inhibited trypsin-treated bloodstream forms were prolonged compared to trypsin-treated populations. In this experiment, the first procyclic forms could be detected on day 7 after initiation of the culture and transformation was complete within 15 days. The transformation of T. b. gambiense from bloodstream to procyclic forms required the living GA-1 cells as feeder layer cells, but established cultures of procyclic forms could be maintained in the culture medium without feeder cells for more than 300 days.

Animals↗

Polyclonal B-cell stimulative and immunosuppressive activities at different developmental stages of Trypanosoma gambiense.

Bloodstream trypomastigote and cultured procyclic (insect midgut) forms of a monomophic strain of Trypanosoma gambiense were tested for their abilities to induce polyclonal B-cell activation (PBA) and immunosuppression (IS) in mice. Injection of a cell homogenate of bloodstream trypomastigotes induced both PBA and IS, while neither PBA nor IS was observed in mice injected with a cell homogenate of cultured procyclics. The results indicate that the substance(s) inducing PBA or IS is related to the developmental stage of the parasites.

Animals↗

Urinary iodide excretion measured with an iodide-selective ion electrode: studies on normal subjects of varying ages and patients with thyroid diseases.

As a part of studies concerning clinical application of the measurement of urinary iodide with an iodide-selective ion electrode, we report here the reference values for the iodide content or concentration in morning spot urine specimens from normal subjects of varying ages and studies with patients with thyroid diseases in Japan. The number distribution of the iodide content or concentration in the morning specimens appeared logarithmic normal in adults, children and infants. Normal ranges found in 95 per cent of populations of healthy subjects were 5.3 to 62.0 mumoles/g Creatinine (Cr) for adults, 5.3 to 42 mumoles/g Cr for children, and 1.9 to 56 microM for infants, respectively. Urinary iodide concentrations in breast-fed infants varied widely compared with those in bottle-fed infants. Mean values were 16.4 microM for breast-fed infants and 8.6 microM for bottle-fed infants, and they were not statistically different. Urine samples from the infants with transient hypothyroidism, who had undergone amniofetography showed extraordinarily high iodide concentrations, even though they were measured at 20th, 29th and 30th days after birth. Although urinary iodide excretion in patients with simple goiter was within normal limits, the mean was statistically lower than that in normal controls (p less than 0.001). Because of the simplicity and rapidity of the electrode method, we strongly recommend it for use in examining iodide excretion in patients with various thyroid diseases.

Adolescent↗

A four generation study of familial dysalbuminemic hyperthyroxinemia: diagnosis in the presence of an acquired excess of thyroxine-binding globulin.

We have studied the largest kindred with familial dysalbuminemic hyperthyroxinemia (FDH) thus far reported, comprising thirty-three blood relations in four generations and three of their spouses. Our objective was to complement previous evidence concerning the precise mode of inheritance of FDH and to detect any other features of the disorder that had not yet been noted. Among the thirty three, there were thirteen patients with FDH, eight males and five females, in all of whom the abnormality appeared to be fully expressed. Within the kindred, no affected female has borne a female child, but transmission from female to male, male to male, and male to female has been observed. Among the offspring of individuals with FDH, the overall observed frequency of FDH in three filial generations was 12/22, or 54.5 per cent, yielding a computed penetrance ratio of 1.09. Four of the patients with FDH had been investigated for hyperthyroidism, and two of them had mistakenly been treated. Of particular interest were two women with FDH who were receiving oral contraceptives and whose serum thyroxine-binding globulin (TBG) concentrations were increased. The results of their thyroid function tests differed from those of patients with FDH whose TBG concentrations were normal and mainly suggested the presence of only a high TBG state. The diagnosis of FDH in these two patients was obscured, and probably would not have been made were it not for the present investigation, which led to the electrophoretic demonstration of increased binding of T4 by serum albumin.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗

A new type of albumin with predominantly increased binding affinity for 3,3',5-triiodothyronine in a patient with Graves' disease.

A new type of serum albumin, that shows a markedly enhanced binding activity for 3,3', 5-triiodothyronine (T3), a somewhat increased activity for thyroxine (T4), and a normal activity for 3,3', 5-triiodothyronine (rT3) is described. This albumin was found in a patient with Graves' disease. After successful subtotal thyroidectomy, the existence of abnormal binding activity for T3 was suspected in this patient because of persistently increased total T3 concentrations in spite of elevated thyrotropin levels. Although free T3 and T4 concentrations measured by radioimmunoassay using commercial tracer analogue kits were markedly increased, those measured by equilibrium dialysis were within normal ranges. Electrophoretic studies revealed that these abnormalities were due to the markedly increased T3 binding activity by the serum albumin; that for T4 was also slightly increased. Scatchard plot analysis revealed that the association constant (Ka) for T3 of the patient's albumin was 5.1 X 10(6)/M (normal pooled albumin; 6.2 X 10(5)/M), and those for T4 and rT3 were 5.2 X 10(6)/M and 2.7 X 10(6)/M, respectively (normal pooled albumin; 2.1 X 10(6)/M for both T4 and rT3). The increased binding of albumin to T3 and T4 was markedly inhibited by barbitone, and 8-anilino-1-naphthalene-sulfonic acid. These characteristic features, and erroneously high values of free T3 and T4 concentrations measured by tracer analogue kits were similar to those seen in patients with familial dysalbuminemic hyperthyroxinemia, which have been previously reported. These findings strongly suggest that this albumin is a new variant in various dysalbuminemic syndromes, and the abnormal binding of iodothyronines moieties in these syndromes are not biochemically identical.

Adult↗

Decrease of free thyroxin in serum of lactating women.

We measured thyroid hormones and thyroxin-binding proteins in serum from 62 normal lactating and 52 nonlactating women at three months postpartum, and compared these values with those for 42 nonpregnant control women of similar age. Mean thyroxin concentrations in the lactating and nonlactating women were significantly (P less than 0.001) lower than that of the nonpregnant controls, but there was no significant difference (P greater than 0.2) in triiodothyronine concentration among these three groups. Free T4 concentration was significantly (P less than 0.01) lower in lactating women than in controls. The reverse-T3 concentrations in both lactating and nonlactating women were significantly (P less than 0.001) lower than in controls, and were significantly (P less than 0.001) lower in lactating than in nonlactating women. The concentration of thyroxin-binding globulin was significantly higher in lactating women than in controls, and the albumin concentration was significantly lower in women postpartum than in controls. Evidently, regulation of thyroid hormone in women postpartum, especially during lactation, differs from that in nonpregnant women.

Adult↗

Contribution of the complement system to antibody-mediated binding of Trypanosoma gambiense to macrophages.

The role of complement in the process of binding of trypanosomes to macrophages in the presence of specific antibody was studied. The aggregation of trypanosomes observed at the optimal antigen-antibody ratio or in the presence of excess antigen inhibited the binding. Complement caused clumped trypanosomes to dissociate, and the free trypanosomes, which were presumed to be coated with antibody that had fixed complement, readily attached to surfaces of phagocytes. Thus, complement was shown to contribute at the site of the antigen-antibody reaction to the creation of an environment suitable for the binding. It seems likely that the trypanosomes dissociated by complement adhered to C3 receptors of the macrophage. However, in the absence of complement and in regions of antibody excess, free trypanosomes also attached to phagocytes. Thus phagocytes may also have receptors for the Fc portion of aggregated antibody. Complement activated by the alternate pathway also enhanced attachment of trypanosomes to phagocytes, but the effect was not as rapid as it was when complement was activated by classical means.

Animals↗

Measurement of iodide in urine using the iodide-selective ion electrode.

A simple and rapid way to measure the concentration of iodide in urine with an iodide-selective ion electrode was described. Potentiometric equilibrium was attained in less than 5 min, and a linear calibration curve was obtained over the potassium iodide (KI) concentration range of 10(-2) to 10(-6) M. The coefficients of variation ranged from 6.2 to 10.0% within assay, and 5.4 to 14.4% between assays. The serial dilution of 3 urine samples with different concentration of iodide showed good linear correlations passing through zero. In practice, the chloride ions in urine did not cause serious errors in the measurement of iodide at molar ratios of chloride ion to iodide up to 2 X 10(4). A good linear correlation was obtained between iodide concentrations in urine determined by the electrode method and by the conventional chemical method (r = 0.92). A linear correlation was also observed between the iodide concentrations of 24 h collected urine and those of single morning urine (r = 0.91). The normal iodide content in single morning urine specimens from 127 Japanese people was 5.3 to 62.0 X 10(-6) moles/g creatinine.

Adult↗

The successful system in long-term cultivation of Trypanosoma gambiense bloodstream forms.

Rat astroglioma cell line (GA-1) was extremely useful for long-term in vitro cultivation of Trypanosoma gambiense blood-stream forms. Parasites could be continuously grown at 37 degrees C for more than 200 days in the culture system, consisted of HEPES-buffered RPMI 1640 (pH 7.2, 300 milliosmole/kg) supplemented with 20% inactivated fetal calf serum in the presence of GA-1 cells. Parasites cultured for more than 200 days still retained not only their virulence for mice but also their original antigenic type. Morphologically, they resembled host infected bloodstream forms by way of having a subterminal kinetoplast and surface coat. The best growth rate of trypanosomes was obtained with 1 X 10(6) GA-1 cells/25 cm2 culture flask. Under this culture condition trypomastigote form populations increased in number up to 7-8 X 10(6) trypanosomes/ml by day 3 after initiation of the culture. The population doubling time in this culture system within the first 24 hours was almost the same as in mice. Most of the cultured trypanosomes were in suspension, but 15-20% of the parasites adhered to the surface of GA-1 cells. The culture system was also shown to be useful for cloning of T. gambiense which is important for separation of mutants.

Animals↗

Heterogeneity of thyroxine binding by serum albumins in normal subjects and patients with familial dysalbuminemic hyperthyroxinemia.

The nature and properties of the T4-binding albumins in the sera of normal subjects and patients with the syndrome of familial dysalbuminemic hyperthyroxinemia (FDH) were investigated by means of isoelectric focusing in polyacrylamide gels. Albumins isolated from normal sera and sera of patients with FDH displayed multiple protein bands, with isoelectric points between 4.65 and 5.75, and protein patterns were the same in the two groups. In normal albumin, [125I]T4 was consistently localized in two bands, termed B1 and B2, whose isoelectric points (pIs) were 5.44 +/- 0.03 and 5.31 +/- 0.02 (mean +/- SE), respectively. Occasionally, binding of far smaller proportions of [125I]T4 was seen in two additional bands of lower pI (5.22 +/- 0.03 and 4.97 +/- 0.07), termed B3 and B4, respectively. In FDH albumin, [125I]T4 was also localized in four bands whose PIs were almost identical to those of B1-B4 in normal albumin. In FDH albumin, however, bands corresponding to B1 and B2 bound only a minor fraction of [125I]T4, the great majority being bound by bands corresponding to B3 and B4, especially the latter. Identity of the four T4-binding albumins in normal and FDH albumin was suggested by their virtually identical pIs in both types of albumin, by the emergence or intensification of the B3 band in both after extraction of endogenous T4, and by the finding that an 125I-labeled derivative of T4 used in a commercial one-step assay for serum free T4 was bound by the B4 band in both normal and FDH albumin, though more intensely in the latter. Treatment of FDH albumin with increasing concentrations of dithiothreitol (DTT; 0.1-5.0 mM) caused a progressive loss of [125I]T4 binding by B3 and B4, leaving [125I]T4 bound by B1 or B2, and had little effect on the binding of [125I]T4 by B1 and B2 in normal albumin. These effects of DTT appeared to correlate well with earlier dialysis studies at physiological pH which revealed that the same concentrations of DTT decreased or abolished the high affinity binding of T4 in FDH albumin but had little effect on the residual lower affinity binding of T4 in FDH albumin or that characteristic of normal albumin. These findings suggest that T4 binding patterns evident during isoelectric focusing of albumin at low pH have relevance to binding of T4 by albumins at physiological pH.(ABSTRACT TRUNCATED AT 400 WORDS)

Alkylation↗

Phospholipase C activity in egg yolk sac of Japanese quail during embryonic development.

1. The amount of lipid decreased markedly at the later stage of embryonic development, coinciding with the remarkable decrease in yolk weight. 2. Phospholipase C activity in the yolk sac appeared on the 6th day of incubation and increased markedly as incubation proceeded. 3. The enzyme showed its optimum activity at pH 9.5 and at 40 degrees C. 4. The activity was enhanced 2-fold by 20 mM CaCl2 and by 40% by 4 mM deoxycholate but inhibited by 2 mM ZnCl2 and CuSO4.

Animals↗

Elevated levels of circulating carcinoembryonic antigen in hypothyroidism.

The level of circulating carcinoembryonic antigen (CEA) was measured in 148 patients with various thyroid diseases. A significantly high frequency of positive CEA was observed in hypothyroid patients with Hashimoto's disease, but the serum CEA levels were not correlated with the serum calcitonin concentrations in Hashimoto's disease. The CEA levels were inversely correlated with the serum T4 concentrations (P less than 0.001) and were positively correlated with the serum TSH concentrations (P less than 0.001), but not with the titers of serum antithyroid antibodies or the size of goiter in autoimmune thyroid disease. Moreover, the increase in CEA was significantly related to the duration of hypothyroidism (P less than 0.001). The high CEA levels in all hypothyroid patients decreased when the patients attained a euthyroid state after thyroid hormone therapy for 4-9 months. The gradual decrease in the serum CEA levels during treatment was roughly correlated with the decreases in serum cholesterol concentration and serum lactic dehydrogenase and glutamic oxalacetic transaminase activities. On Sephadex G-200 column chromatography of serum from hypothyroid patients, the CEA immunoreactivity, like purified standard CEA, was recovered in the large molecular weight fraction. These findings indicate that elevated CEA levels in hypothyroid patients do not necessarily indicate malignancy. CEA elevation in hypothyroidism may be caused by decreased degradation of CEA.

Adolescent↗

Serum ratio of triiodothyronine to thyroxine, and thyroxine-binding globulin and calcitonin concentrations in Graves' disease and destruction-induced thyrotoxicosis.

The serum ratios of T3 to T4, and T4-binding globulin (TBG) and calcitonin concentrations were studied in cases of thyrotoxic Graves' disease and destruction-induced thyrotoxicosis. In 272 patients with Graves' disease, 209 of 240 (87%) untreated patients without complications had high T3 to T4 ratios (nanograms per micrograms) of more than 20. Six of 32 (19%) patients with Graves' disease who had complications (15 with pregnancy, 14 with increased TBG, and 3 with conditions associated with a low T3 syndrome) had high T3 to T4 ratios. Eleven of 74 (15%) patients with destruction-induced thyrotoxicosis (24 with subacute thyroiditis, 39 with postpartum transient thyrotoxicosis, and 11 with spontaneous transient thyrotoxicosis) had high T3 to T4 ratios. Patients who had serum T4 levels of more than 30 micrograms/dl and/or T3 levels of more than 800 ng/dl had Graves' disease. There was no significant correlation between the T3 to T4 ratio and activities of thyroid-stimulating immunoglobulins in thyrotoxic patients with Graves' disease who had no complications. The average serum levels of TBG in destruction-induced thyrotoxicosis and thyrotoxic Graves' disease were 20.7 +/- 4.3 micrograms/ml (mean +/- SD; n = 22), and 19.9 +/- 4.0 (n = 41), respectively, which were significantly lower than that in healthy subjects (22.7 +/- 4.4 micrograms/ml; n = 165), but there was no difference between the values in the two groups of thyrotoxicosis patients. The average serum level of calcitonin in destruction-induced thyrotoxicosis patients was 96.7 +/- 66.7 pg/ml (n = 21), which was significantly (P less than 0.05) higher than the values in patients with thyrotoxic Graves' disease (62.0 +/- 44.7 pg/ml; n = 26) and in healthy subjects (63.9 +/- 31.2 pg/ml; n = 29), but the difference in values in the two groups of thyrotoxicosis was not clinically useful because of considerable overlap of individual values. The T3 to T4 ratio is a simple and helpful index for the differentiation of the two types of thyrotoxicosis. A T3 to T4 ratio less than 20 in thyrotoxic patients before therapy is a laboratory signal of destruction-induced thyrotoxicosis or Graves' disease with complications, but final differentiation should be confirmed by measuring radioactive iodine uptake.

Adult↗