"Normal suppression" to dexamethasone in Cushing's disease: an expression of decreased metabolic clearance for dexamethasone.

The adrenal cortical function of a patient with pituitary-dependent Cushing's syndrome exhibited normal responsiveness to conventional doses of dexamethasone (Dex) over several years of evaluation. "Periodic hormonogenesis" did not seem to explain the phenomenon. Plasma concentrations of Dex were measured to ascertain whether an abnormality in Dex metabolism might explain the apparent discrepancy in Dex responsiveness. Plasma levels of Dex after oral administration of the steroid were higher than normal, suggesting that decreased clearance of Dex accounts for the phenomenon of "normal suppression" in this patient with Cushing's syndrome.


Effect of indomethacin on the hypothalamic-pituitary-adrenal axis in man.

The effect of the cyclo-oxygenase inhibitor, indomethacin, on hypothalamic-pituitary-adrenal function was investigated in five normal males. Baseline 0800 and 1600 h plasma ACTH and cortisol, plasma ACTH and 11-deoxycortisol responses to metyrapone, and plasma cortisol responses to dexamethasone and exogenous ACTH were not affected by indomethacin. However, the area under the curve for plasma ACTH after iv injection of regular insulin (0.1 U/kg) was significantly decreased during indomethacin administration (control, 88.0 +/- 32.6 cm2, mean +/- sd; indomethacin, 47.6 +/- 23.1, P less than 0.01). Plasma cortisol levels were decreased only at 30 min. Our results support the hypothesis that prostaglandins or any of their precursors play a role in the hypothalamic control of ACTH secretion and indicate that evaluation of hypophyseal ACTH secretory capacity by means of insulin-induced hypoglycemia may yield abnormal results in patients receiving indomethacin.


Formation of 2- and 4-hydroxyestrogens by brain, pituitary, and liver of the human fetus.

When [6,7-3H]estradiol was incubated with tissue homogenates of the brain, the pituitary, and the liver of two human female fetuses, a number of radioactive metabolites more "polar" than the incubated substrate were detected. Among these, the identification of two types of catecholestrogens, i.e. the 2- and 4-hydroxyestrogens, was of major interest. Compared on the basis of wet weight of tissues (250 mg), the conversion of estradiol to 2-hydroxyestrogens (2-hydroxyestradiol and 2-hydroxyestrone) was 0.8% in the frontal cortex, 1.0% in the hypothalamus, 2.1% in the pituitary, and 7.8% in the liver. For the first time, the formation of 4-hydroxyestrogens was demonstrated. The percentages of incubated estradiol hydroxylated at C-atom 4 (4-hydroxyestradiol and 4-hydroxyestrone) were 0.5 in the cortex, 0.4% in the hypothalamus, .1% in the pituitary, and 0.5% in the liver. The results show that fetal brain and pituitary tissue can hydroxylate estradiol in positions 2 and 4 to a similar extent, whereas in the liver, about 15 times more 2-hydroxy than 4-hydroxy compounds are formed. Moreover, the 2-hydroxylating capacity of the liver is definitely greater than that of the brain, whereas the 4-hydroxylating capacity is about the same as that of the brain.


Pathophysiological studies in idiopathic hypercalciuria: use of an oral calcium tolerance test to characterize distinctive hypercalciuric subgroups.

Twenty-one unselected patients with recurrent nephrolithiasis and normocalcemic hypercalciuria with or without hypophosphatemia and 18 normal subjects were studied with an oral calcium tolerance test and for 3- to 5-day periods while consuming a low normal (400 mg) and high-normal (1000 mg) calcium intake. The oral calcium tolerance test consisted of the measurement of the calcemic, calciuric, and parathyroid (assessed by determinations of serum immunoreactive parathyroid hormone and nephrogenous cAMP) responses to acute 1000- or 350-mg doses of calcium. Nineteen patients displayed normal results for basal serum calcium, parathyroid function, and fasting calcium excretion, and striking calcemic (mean increase in serum calcium, 0.9 vs. 0.2 mg/dl in the normal subjects) and calciuric (mean increase in urinary calcium, 0.33 vs. 0.15 mg calcium/100 ml GF in the normal subjects) responses to the 1000-mg calcium tolerance test, associated with a mean 54% suppression in nephrogenous cAMP. These patients were operationally defined as having "absorptive" hypercalciuria. The variable occurrence of hypophosphatemia in this group suggested that the pathogenesis of "absorptive" hypercalciuria may be complex and/or multifactorial. There were strong positive correlations between the calciuric response to the calcium tolerance test and fractional isotopic calcium absorption (r = 0.75, P less than 0.00), the calcemic responses to the test (r = 0.71, P less than 0.001) and the calciuric responses noted on the 1000- vs. the 400-mg daily calcium intake (r = 0.78, P less than 0.001). Two patients displayed low or low normal basal serum calcium, increased parathyroid function, increased fasting calcium excretion, and a striking calciuric but minimal calcemic response to the 1000-mg calcium tolerance test, associated with a moderate suppression in nephrogenous cAMP. These patients were operationally defined as having "renal" hypercalciuria. Several lines of evidence indicated that the hyperparathyroidism in these patients was physiological or secondary, including the near normalization of parathyroid function on the daily 1000-mg calcium intake. A steep slope of calcium excretion on calcium intake (due to increased calcium absorption) was noted in all hypercalciuric patients and accounted for the significantly improved diagnostic accuracy of screening patients for hypercalciuria on the high-normal calcium intake. The simple measurement of total cAMP excretion (nanomoles per 100 ml GF) and urinary calcium on the 1000-mg daily calcium intake seemed to provide reliable separation of patients with "renal" from those with "absorptive" hypercalciuria. A physiological (350 mg) dose of oral calcium produced a 30% suppression of nephrogenous cAMP in normal subjects; this suggests that dietary calcium exerts an important control of parathyroid function under physiological circumstances.


Cortisol and androgen secretion in a case of Nelson's syndrome with paratesticular tumors: response to cyproheptadine therapy.

Bilateral paratesticular tumors were observed in a 32-yr-old man 14 yr after he developed a pituitary tumor after adrenalectomy for Cushing's disease (Nelson's syndrome). Plasma ACTH concentrations were markedly elevated (mean, 6350 pg/ml), but urinary free cortisol concentrations were low (27-31 micrograms/24 h). Catheterization revealed a spermatic to peripheral venous gradient for cortisol consistent with secretion of this steroid by the tumor. This was confirmed by decreased cortisol excretion after tumor excision. Serum androgen (testosterone, androstenedione, dihydrotestosterone, and dehydroepiandrosterone-sulfate) and progestin (progesterone and 17-hydroxyprogesterone) concentrations were decreased and did not decline further after tumor removal. These latter observations suggested that the paratesticular tumors did not secrete appreciable testosterone or any of its immediate precursors. Serum gonadotropin levels were also low. Cyproheptadine treatment resulted in a marked lowering of plasma ACTH concentrations (221-320 pg/ml). This was associated with an increase in both plasma LH and testosterone concentrations. These observations are consistent with the hypothesis that ACTH may directly affect LH and testosterone secretion.


Urinary free 18-hydroxy-11-desoxycorticosterone excretion in normal and hypertensive patients.

One hundred fourteen hypertensives and 20 normal controls were examined using a new clinical technique of measuring 24-h urinary free 18-hydroxy-11-desoxycorticosterone (18-OH-DOC) excretion in response to dietary salt manipulations and ACTH injections. The object was to avoid potential errors of random plasma sampling. Mean urinary free 18-OH-DOC in normals on 110 milliequivalent sodium diet was 1.84 +/- 0.69 microgram (mean +/- SD) and represented about 2% of the daily secretion rate of this steroid. Both in normals and hypertensives, urinary free 18-OH-DOC approximately doubled on low salt (P less than 0.01 for each) and rose about 10 times in response to ACTH injection (P less than 0.05 and P less than 0.01, respectively). Plasma and urinary free 18-OH-DOC showed good correlation in patients with essential hypertension on a low salt diet (r = 0.45, P less than 0.01). Suppressed renin patients showed no propensity toward excess 18-OH-DOC excretion and hypertensives with elevated 18-OH-DOC could not be distinguished by their aldosterone levels, cortisol levels, nor their responses to various stimuli. These data suggest 18-OH-DOC is predominantly secreted under ACTH control and, to a smaller extent, in response to salt changes. Hypertension characterized by chronic overproduction of 18-OH-DOC forms only a small percentage of the hypertensive population. It is proposed that measuring 24-h urinary free 18-OH-DOC excretion may be the best method of assessing its rate of secretion without resorting to injection of radiolabeled material.


On the isolation of human pituitary hormones.

Six human pituitary hormones can be obtained from a single batch of human pituitary glands by the procedure described. Starting from an acetone powder of the glands, the hormones are segregated into three fractions; one containing GH and PRL, a second containing the glycoprotein hormones, and the third fraction containing smaller peptides including ACTH and MSH. Clinical grade GH is extracted from the residue at alkaline pH, followed by a series of ammonium sulfate and pH fractionations. Further chromatography on G-100 yields a highly purified GH preparation. FSH is purified by chromatography on carboxymethyl cellulose at pH 5.4 followed by gel filtration on G-100 and fractionation on sulfopropyl Sephadex C-50 at pH 5.4. LH and TSH are separated by DEAE-cellulose chromatography at pH 9.5. The GH and glycoprotein hormones are recovered in a highly purified form and good yields. The GH is obtained in an active form which is easily soluble and well suited for clinical use. A large part of the PRL is also saved during the extraction procedure.


Plasma parathyroid hormone and calcium are related to sleep stage cycles.

To study dynamic interactions among parathyroid hormone (PTH), plasma calcium, and brain states, seven normal subjects were studied for a total of eight nights in our sleep laboratories. Plasma samples were obtained at 10- to 20-min intervals for PTH and calcium determinations. Electroencephalogram, eye movements, and muscle tone were recorded to determine sleep stages. On each night, several distinct peaks in PTH concentration were seen, which in some cases exceeded the all night mean PTH by as much as 300%. Peaks in plasma PTH were significantly nonrandom and tended to recur about every 100 min. PTH concentration was significantly related to cycles of stages 3 and 4 sleep. Total plasma calcium varied less but was significantly related to cycles of rapid eye movement sleep and to cycles of stage 2 sleep. PTH and calcium were significantly interrelated, especially at high frequencies above 40 cycles/day (1 cycle 36 min). In the 14.4 cycles/day (1 cycle/100 min) frequency range where most PTH and calcium variability was found, however, PTH and calcium were more closely related to sleep stages than to each other. These results suggest that the regulation of PTH and calcium is complex and may involve interactions with neural systems.


Treatment of Cushing's syndrome with trilostane (WIN 24,540), an inhibitor of adrenal steroid biosynthesis.

Seven patients with Cushing's syndrome were treated with trilostane (WIN 24,540) 4 alpha,5-epoxy-17 beta-hydroxy-3-oxo-5 alpha-androstane-2 alpha-carbonitrile), an inhibitor of adrenal steroid biosynthesis. Trilostane treatment reduced steroid biosynthesis and it also improved biochemical manifestations of the disease in all of the patients treated. The average cortisol secretory rate decreased significantly with treatment, from 47.1 to 23.4 mg/24 h (P less than 0.005), and urinary 17-hydroxycorticosteroids decreased from 15.7 to 8.7 mg/24 h (P less than 0.01). Urinary free cortisol excretion decreased from 277 to 88 microgram/24 h (P less than 0.01), and 0800 h plasma cortisol levels declined from 25.0 to 12.0 microgram/dl (P less than 0.05). Conversely, dehydroepiandrosterone sulfate excretion in urine increased from 1.3 to 5.8 mg/24 h (P less than 0.0025) and in plasma increased from 162 mg/24 h (P less than 0.025). Plasma and urinary free dehydroepiandrosterone increased 2-fold. Urinary 17-ketosteroid excretion increased from 18 to 43 mg/24 h (P less than 0.001). A significant reduction in urinary excretion of tetrahydroaldosterone, tetrahydrodeoxycorticosterone, and 18-hydroxytetrahydrodeoxycorticosterone was observed with treatment. Inhibition of steroid biosynthesis was accompanied by a 2-fold increase in PRA and no change in serum cholesterol levels. Mean arterial blood pressure decreased with treatment from 109 to 97 mm Hg (P less than 0.005), and fasting blood sugar decreased from 117 to 98 mg/dl (P less than 0.005), accompanied by rise in plasma potassium levels from 3.8 to 4.3 milliequivalents/liter (P less than 0.025). Two patients on long term therapy also showed an improvement in clinical features of their disease. There were no significant treatment-related carcinoma, simultaneously producing both an excessive amount of cortisol and ACTH, is described. It is concluded that trilostane is an effective inhibitor of 3 beta-hydroxysteroid dehydrogenase enzyme system in human adrenal gland; it inhibits biosynthesis of cortisol and it is useful in the treatment of Cushing's syndrome.


Effects of thyroid-stimulating hormone on human thyroid carcinoma and adjacent normal tissue.

Effects of TSH on the adenylate cyclase-cAMP system and some parameters of intermediary metabolism were investigated in human thyroid carcinoma and adjacent normal thyroid tissue. Basal adenylate cyclase activity and cAMP concentrations were significantly higher in carcinomatous tissue. Basal [1-14C]glucose oxidation, 32Pi incorporation into phospholipids, and organification of iodide were similar in both tissues. Stimulation of cAMP by TSH was significantly greater in normal compared to carcinomatous tissue. In neither tissue was there a good correlation between TSH stimulation of adenylate cyclase activity and cAMP concentrations. The TSH stimulation of 32Pi incorporation into phospholipids by TSH was significantly greater in normal tissue. The mean effect of TSH on iodide organification and glucose oxidation was similar in normal and carcinomatous tissue. Although specific binding of TSH was demonstrated in both normal and carcinomatous tissue, it did not correlate very well with stimulation of adenylate cyclase activity. Hormones other than TSH also augmented adenylate cyclase activity in two of the carcinomas. In individual patients, the relative responsivity of carcinomatous tissue compared to normal was not always consistent when all of the metabolic parameters were considered.


Alteration of feedback mechanism of estrogen on gonadotropin by sulpiride-induced hyperprolactinemia.

Four normally cycling women received an iv injection of 20 mg Premarin (conjugated estrogens equivalent to 20 mg estrone sulfate) on the seventh day of two consecutive cycles; the second experiment was performed under sulpiride-induced hyperprolactinemia (mean PRL level = 906 microU MRC standard 71/222 per ml; significantly 7.8 times greater than mean control level of 115 microU MRC standard 71/222 per ml, P less than 0.001). In comparison to the control experiment, sulpiride-induced hyperprolactinemia prevented the occurrence of any gonadotropin peak within the 84 h of estrogen administration; the negative feedback effect of estrogen on gonadotropin secretion was maintained and was even potentiated. These alterations of feedback mechanisms of estrogen were considered to be related to hyperprolactinemia itself rather than to sulpiride. Five other normally cycling women received iv injections of 100 microgram LRH on the 22nd day of a cycle under sulpiride-induced hyperprolactinemia since the onset of menstruation. Their mean LH response was somewhat greater (although not statistically significant) and their mean FSH response was considerably greater (P less than 0.001 at all times) than those of a control group of 10 women tested in their luteal phase. The results of these LRH tests under sulpiride-induced hyperprolactinemia give some support to the concept that hyperprolactinemia interferes at the hypothalamic or higher level with cyclic release of indogenous LRH.


Comparison of absorption of cortisone acetate and hydrocortisone hemisuccinate.

In four patients who required maintenance glucocorticoid therapy after bilateral adrenalectomy for Cushing's disease, we compared the effects of im injection and oral ingestion of cortisone acetate and hydrocortisone hemisuccinate. By the former route of administration, cortisone acetate was not effective in elevating plasma cortisol levels or in suppressing plasma adrenocorticotropin, although hydrocortisone was. When given by mouth, no significant difference was found between the two steroids. Therefore, in the treatment of acute adrenal insufficiency or in the maintenance of patients with chronic adrenal insufficiency and in their preparation for surgery or other stressful situations, we advise against im injection of cortisone acetate. Oral ingestion, however, is appropriate for maintenance.


Age-related response of plasma testosterone, delta 4-androstenedione, and cortisol to adrenocorticotropin in infants, children, and adults.

The response of plasma testosterone (T), delta 4-androstenedione, and cortisol (F) to the administration of synthetic Zn beta 1-24 ACTH (0.5 mg/m2 im every 12 h for 3 days) was ascertained in 20 infants, 35 prepubertal children, 4 early pubertal boys, and 15 adults. At all ages and in both sexes, a significant rise in delta 4-androstenedione and F was observed (P less than 0.00001), whereas the response of T showed a sex difference: T levels increased in females (P less than 0.001) at all ages in response to ACTH, while they decreased (P less than 0.01) in males at periods of active testicular secretion (early infancy, puberty, and adulthood). In prepubertal boys, in the absence of significant Leydig cell activity, T levels increased after ACTH, as they did in girls. The post-ACTH values of T, expressed as percent-ages of the control levels, were significantly lower (P less than 0.01) in infants (3.17 +/- 16.7%) than in pubertal boys (59.5 +/- 14.6%) or adult men (57.9 +/- 7.7%). F levels were significantly higher after ACTH stimulation at 1-4 months of age (253 +/- 129 micrograms/dl) than at any later age studied (64 +/- 17 micrograms/dl). It would seem, therefore, that the suppressive effect of ACTh on testicular secretion might be glucocorticoid mediated and its magnitude might be related to circulating levels of F.


Circulating parathyroid hormone activity: familial hypocalciuric hypercalcemia versus typical primary hyperparathyroidism.

Three indices of circulating parathyroid hormone (PTH) activity were compared between two groups: the first a group of 23 patients from three large kindreds with autosomal dominant hypercalcemia without hypercalciuria [familial hypocalciuric hypercalcemia (FHH)] and the second a group of 64 patients with typical primary hyperparathyroidism (1HPT) manifesting comparable hypercalcemia. The group with 1HPT differed from normal with respect to plasma PTH 1HPT concentration (normal, less 0.2 ng/ml), urinary cAMP excretion per 100 ml glomerular filtrate (U cAMP/GF) (normal, 2.3 x/divided by 0.6 nmol/100 ml glomerular filtrate; mean, x/divided 1 SD), and renal tubular maximum of phosphate transport corrected for glomerular filtration rate (TMP/GFR; normal, 3.4 +/- 0.4 mg/dl; mean, +/- 1 SD). The group with 1HPT also diverged significantly from the group with FHH for all three indices: for PTH, 0.37 x/divided by .48 vs. 0.25 x/divided .46 (P less than 0.05); for UcAMP/GF, 4.3 x/divided by .53 vs. 2.6 x/divided .60 (P less than 0.0005); and for TMP/GFR, 2.0 +/- 0.6 vs. 2.6 +/- 0.7 (P less than 0.01). The between-group differences for all three indices were also significant after adjustment for their variation with serum calcium. However, only the difference in TMP/GFR remained significant after adjustment for covariance attributable to serum calcium concentration, age, and creatinine clearance. The group with FHH differed from normal for TMP/GFR but not for UcAMP/GF. However, analysis of changes in UcAMP/GF and serum calcium concentration around the time of parathyroidectomy in three patients with FHH suggested that the parathyroid glands contributed to the abnormalities of mineral homeostasis in at least one. It was concluded that higher serum concentrations of PTH do not account for the lower renal clearance of calcium and magnesium in FHH calcium concentration, the group with FHH showed indices suggesting lower circulating PTH activity than the group with 1HPT.


The effect of long acting glucocorticoids on menstrual abnormalities in patients with virilizing congenital adrenal hyperplasia.

Amenorrhea or oligomenorrhea occurs in more than 25% of young women with virilizing congenital adrenal hyperplasia due to 21-hydroxylase deficiency who are treated with conventional glucocorticoid therapy. In four such patients, we studied the variation in plasma concentration of ACTH, 17-hydroxyprogesterone (17-OHP), estradiol (E2), estrone, testosterone (T), dehydroepiandrosterone and its sulfate, androstenedione, and progesterone before and at frequent intervals after the administration of the patients' usual dose of oral glucocorticoids. We found that plasma ACTH, 17-OHP, and T were usually increased above the normal range early in the morning before the first daily dose of medication; plasma dehydroepiandrosterone and its sulfate were decreased below the normal range for the patient's pubertal stage; and androstenedione, estrone, and E2 were within the normal range. Two of the four patients had low basal plasma LH and impaired LH response to LRF. Administration of a long acting glucocorticoid (dexamethasone) to three patients, and methylprednisolone to one, resulted in continuous suppression of T into the normal range in all patients, in spite of continued abnormally increased concentration of ACTH and 17-OHP in two patients. In three patients, E2 concentrations were increased when a long acting glucocorticoid was given. In all four patients, menses began or became more regular during treatment with the long acting glucocorticoid. We conclude that a long acting glucocorticoid is useful in the management of menstrual abnormalities in adolescent patients with congenital adrenal hyperplasia  who have attained their adult height, and that monitoring the concentration of serum T in them is a valuable but not infallible procedure for assessing the effectiveness of therapy.


Abnormalities of triiodothyronine binding to lymphocyte and fibroblast nuclei from a patient with peripheral tissue resistance to thyroid hormone action.

T3 binding to lymphocyte nuclei has been studied in normal individuals and in a patient (MaG) with peripheral resistance to thyroid hormone action. This syndrome is defined by the presence of hypothyroidism or euthyroidism with high plasma levels of thyroid hormone. T3 bound to a single set of binding sites in normal adult lymphocyte nuclei with a mean Ka of 8.9 +/- 7.1 x 109 M-1, and a capacity of 4.4 +/- 2.9 fmol/100 micrograms DNA. A single binding site was also disclosed in MaG's lymphocytes with a Ka of 0.43 x 109 M-1 and a capacity of 10.5 fmol/100 micrograms DNA. This low affinity was not due to the presence of high plasma T3 level in the patient, since administration of 100 micrograms T3 to normal adult volunteers induced the presence of two different binding sites. The mechanism responsible for this phenomenon is unknown. To binding was also studied using cultured fibroblasts which were incubated in serum-less medium before the binding experiments. One single binding site (Ka, 1.9 x 10(10) M-1, capacity, 12.9 fmol/100 micrograms DNA) was detected in normal fibroblast nuclei. In contrast, a curvilinear Scatchard plot was obtained when MaG's fibroblasts were used. This result could be compatible with the presence of either two different binding sites or negative cooperativity. In support of the latter possibility, Hill plots gave a number lower than unity. The results suggest that the syndrome of peripheral tissue resistance to thyroid hormone action due to a defect at the level of the nuclear receptor. The possible existence of similar syndromes due to an alteration at the level of a post-T3-binding mechanism is not eliminated.


A familial syndrome of decrease in sensitivity to 1,25-dihydroxyvitamin D.

Typical features of hereditary vitamin D-dependent (pseudovitamin D-deficient) rickets were observed beginning at ages 20 and 5 months in a brother and sister. Both had calcium malabsorption correctable with high doses of 25-hydroxyvitamin D. During periods of hypocalcemia they both manifested secondary hyperparathyroidism with hypophosphatemia and high serum concentrations of endogenously produced 1,25-dihydroxyvitamin D. In each, normalization of serum calcium concentration and resolution of osteomalacia were obtained with continuous administration of high doses of ergocalciferol or high doses of 1,25-dihydroxycholecalciferol. Chemical features of vitamin D deficiency were corrected in the presence of high circulating concentrations of 1,25-dihydroxyvitamin D2, produced endogenously, or of 1,25-dihydroxyvitamin D3, administered by mouth. Serum concentrations of 25-hydroxyvitamin D2, 25-hydroxyvitamin D3, 24,25-dihydroxyvitamin D, and 1,25-dihydroxyvitamin D were normal in five first degree relatives. We conclude that in these five first degree relatives. We conclude that in these siblings, rickets and osteomalacia resulted from a hereditary decreased sensitivity to 1,25-dihydroxyvitamin D at the intestine and perhaps other vitamin D target tissues.


Ectopic production of high molecular weight calcitonin and corticotropin by human small cell carcinoma cells in tissue culture: evidence for separate precursors.

The DMS-79 continuous line of human small cell lung carcinoma cells, which produces immunoreactive (IR)-corticotropin (ACTH), -lipotropin (LPH), and -beta-endorphin (beta END), was found to produce IR-calcitonin (CT). Two major high molecular weight (HMW) forms of IR-CT were observed after gel exclusion chromatography under denaturing conditions (mol wt. approximately 7,000 and approximately 14,000), as well as a minor HMW IR-CT component (mol. wt. approximately 70,000). None of these IR-CT materials was extracted from DMS-79 medium by affinity chromatography using an ACTH antibody covalently bound to agarose. These results demonstrate ectopic production of HMW forms of CT and ACTH/LPH/beta END by human lung tumor cells in tissue culture, but do not support the existence of a common CT/ACTH/LPH/beta END precursor molecule.


Studies on platelet aggregation inhibitors in vivo. VIII. Effect of pentoxifylline on spontaneous tumor metastasis.

Pentoxifylline (Trental), a phosphodiesterase inhibitor with platelet aggregation inhibitory effect, was shown to decrease spontaneous metastases in a Wilms' tumor model in Furth-Wistar rats and in neuroblastoma C1300 in A/J mice. It was ineffective in the NIH renal adenocarcinoma in BALB/cCr mice.


Niemann-Pick disease: prenatal diagnoses and studies of sphingomyelinase activities.

Five pregnancies were monitored for couples at-risk for having a child with some form of Niemann-Pick disease (NPD). Three of these were for the classic Type A form in which affected children usually have less than 1% of normal sphingomyelinase activity. Two of these pregnancies were terminated after the cultured amniotic-fluid cells were determined to have less than 1% of normal sphingomyelinase activity (0.4 and 0.6 nmole/mg protein/hr versus the control mean of 61.7). In the other pregnancy at risk for Type A NPD near normal activity was measured and it was continued to term. The two other pregnancies were monitored for couples in which severely affected children were found to have partially deficient sphingomyelinase activity (about 20% of normal) in cultured skin fibroblasts. Cultural amniotic-fluid cells from one of these pregnancies also had about 20% of control sphingomyelinase activity, but the woman underwent a spontaneous abortion soon after the cells were received and no studies on the fetus were done. The other sample was taken at the time of abortion for social reasons. In this case the cultured amniotic-fluid cells and cultured fetal skin fibroblasts gave normal sphingomyelinase activity. Enzymatic studies on tissues from the two fetuses predicted to be affected with Type A NPD confirmed the prenatal diagnosis. Studies of sphingomyelinase activity in the brains from these fetuses and from a child who died with Type A NPD indicated significant levels of activity when measured at pH 7.4 in the presence of magnesium. The higher level of the pH 7.4 sphingomyelinase activity in developing brain may indicate some important role in normal brain development.


Pseudohypoparathyroidism and idiopathic hypoparathyroidism: relationship between serum calcium and parathyroid hormone levels and urinary cyclic adenosine-3',5'-monophosphate response to parathyroid extract.

Forty patients with hypocalcemia and/or Albright's hereditary osteodystrophy were studied. Based on the estimation of serum calcium and parathyroid hormone (PTH) levels as well as the urinary cAMP response to infusions with parathyroid extract, it was possible to classify all of the patients studied as cases with idiopathic hypoparathyroidism (n = 6, low PTH, normal cAMP response), pseudohypoparathyroidism (PHP) type I (n = 18, high PTH, low cAMP response) and type II (n = 2, high PTH, normal cAMP response), as well as pseudopseudohypoparathyroidism (n = 14, normal PTH, normal cAMP response). In three cases studied at the age of 12, 17, and 23 yr, the signs of Albright's hereditary osteodystrophy were not observed. PTH levels were unusually high for a given serum calcium concentration in some patients with PHP, the increased PTH levels were, however, normalized during iv calcium infusions. In two young children with PHP, a gradual increase of serum PTH levels occurred despite persistent normocalcemia over a period of 3 yr. This suggests that factors other than hypocalcemia or frequent small unobservable falls of the serum calcium concentration, such as a deficient formation of 1,25-dihydroxyvitamin D3, secretion of an abnormal PTH, or an abnormal metabolism of the hormone, may contribute to the secondary hyperparathyroidism in PHP.


Diurnal rhythm and disappearance half-time of endogenous plasma immunoreactive beta-MSH (LPH) and ACTH in man.

In order first to establish the disappearance t 1/2 of endogenous immunoreactive human beta-MSH (RIA-h beta MSH), which is now known to represent the lipotropins, human beta-LPH and human gamma-LPH, the plasma concentrations of RIA-h beta MSH, RIA-hACTH, and fluorogenic corticosteroids (cortisol) were measured before and during infusion of cortisol in three patients with primary adrenocortical insufficiency from whom steroid replacement had been withdrawn. The t 1/2 for RIA-h beta MSH ranged from 44-133 min (mean, 83), and for RIA-hACTH ranged from 20-51 min (mean, 40). On this basis, 30-min sampling intervals were chosen to measure plasma concentrations of RIA-h beta MSH, RIA-hACTH, and cortisol in three normal subjects for 24 h. Similar diurnal rhythms were observed for all three hormones. Highest values (58.0 +/- 5.8 pg/ml) for RIA-h beta MSH were observed at about the time of awaking and lowest values (15.0 +/- 2.1 pg/ml) shortly after retiring. The secretion of both RIA-h beta MSH and RIA-hACTH appeared to be episodic, with about 8-10 secretory episodes/24 h, and the timing of their secretory episodes appeared to be nearly identical. There was excellent correlation (P less than 0.001) between simultaneous concentrations of RIA-h beta MSH and RIA-hACTH in all three subjects. These results establish the presence of a diurnal rhythm in plasma lipotropins (RIA-h beta MSH) and suggest that roughly equimolar amounts of ACTH and LPH are secreted simultaneously by the pituitary under basal conditions in man.


Cell-free translation of messenger RNA extracted from a human insulinoma.

Total nucleic acid has been extracted from a human pancreatic insulinoma. Purification of the messenger RNA (mRNA) fraction by oligo-dT cellulose chromatography yielded 200 micrograms poly(A)-rich mRNA. This mRNA produced a 2-fold stimulation of protein synthesis in a wheat germ cell-free system. Analysis of the translation products by gel filtration chromatography (Biogel P-30) revealed nothing smaller than an acid-alcohol-soluble protein larger than bovine proinsulin. In contrast, insulinoma slices incubated with labeled amino acids synthesized smaller proteins which comigrated with bovine proinsulin and insulin. [3H]Leucine-labeled cell-free proteins were electrophoresed on NaDodSO4-urea polyacrylamide slab gels. In the presence of insulinoma mRNA, discrete proteins of 25,000 and 11,500 mol wt were synthesized. The 11,500 mol wt protein was specifically immunoprecipitated with antiinsulin serum. Thus, cell-free translation of human insulinoma mRNA yields an immunoreactive insulin larger than proinsulin, which is the same size as fish and rat preproinsulins recently described.


A method for determining bone mineral content using Fourier image reconstruction and dual source technique.

The basic principles of a quantitative tomographic technique for bone mineral determination are considered. The bone mineral content could be evaluated from a single reconstruction using projection data from two different energies. Beam hardening effects and photon counting statistics are studied by computer simulation and found to be the limiting factors in precision and accuracy.


Computed tomographic demonstration of inferior vena cava invasion in a case of hepatocellular carcinoma.

A case of hepatocellular carcinoma with neoplastic invasion of the inferior vena cava is presented. Computed tomograms before and after injection of contrast medium revealed an unusual density enhancement of the wall of the invaded portion of the vena cava. This sign is correlated with an angiographic sign of venous invasion present in our case.


[Morphological characteristics and the degree of somatic maturity in children of short stature].

An auxological investigation involved 199 children with short stature ( hypostatura simplex) in preschool and school age. The analysis of the material showed that eight anthropometrical patterns and five somatometric indices, with the evaluation of the level of skeletal maturity, were all lower in children under study in comparison to the mean growth standard for Warsaw children. In all features differences were statistically significant. The biggest differences however, were found in the length of the upper extremities and the trunk. Observed differences increase with age. The authors concluded that the children under study were harmoniously delayed in morphological age which was confirmed by the level of the index of biological maturity based on skeletal age. It was found that the level of biological maturation was delayed but not to such a great extent as in growth pathology.


[Bilateral nephroblastoma in children].

The rarity of such form of nephroblastoma containing about 4% among embryonal renal tumors in this paper, was the subject of special attention. The necessity of preoperative differential diagnosis particularly in cases of congenital multicystic kidney, congenital hydronephrosis and retroperitoneal sympathetic neoplasms was stressed. Radiological investigation facilitate distinguishing of the three pathological forms. A multicystic degeneration of kidney is usually bilateral and it includes both the medulla and the renal cortex. The cystic patterns nearly equal in size are multiple and minute. In the bilateral Wilms' tumour besides the kidney enlargement there are deformations such as hydrocalyx , hydronephrosis or modeling signs. In case of congenital hydronephrosis the kidney keeps the same size and produces the thinner cortex zone. The neoplasms of the sympathetic nerve origin produce first the displacement of kidney and then the deformation of pelvico -calyceal system with modeling signs and sometimes with hydronephrosis. In the above mentioned entities, late urographical pictures are of great significance. The angiographic investigations of kidney play a very important part in the differential diagnosis and planning of operative procedures. The nephrographic phase of angiography shows the quantity and location of active renal parenchyma. A renal arteriography is also important in the evaluation of the applied therapy, particularly in case of multistage therapeutic procedures.


Supersensitivity to norepinephrine or dopamine antagonists after knife cuts that produce aphagia and adipsia in rats.

Aphagia and adipsia of equivalent duration were produced by knife cuts along the lateral border of the hypothalamus (PH cuts), or the medial surface of the globus pallidus (MP cuts) in male albino rats. Striatal dopamine (DA) was reduced by 75% in animals with PH cuts but only 50% by MP cuts. Hypothalamic norepinephrine was reduced 25% by PH cuts and was unaffected by MP cuts. Aphagia and adipsia were positively correlated with DA depletions only in rats with PH cuts. Presurgical catecholamine depletions produced by chronic injections of alpha-methyl-p-tyrosine did not alter the duration of aphagia or adipsia resulting from these knife cuts. However, following recovery of ingestive behavior, rats with PH and MP cuts were supersensitive to the anorexic effects of the dopamine-beta-hydroxylase inhibitor, diethyldithiocarbamate. Exaggerated anorexia was also observed after DA blockade by haloperidol or alpha-adrenergic blockade by phenoxybenzamine. The most pronounced effects of catecholamine blockade were observed in rats with PH cuts.


Debilitating interaction of adrenalectomy and intrahypothalamic implants of prostaglandin E2 upon open-field activity levels and sexual receptivity in estrogen-primed ovariectomized rats.

A group of estrogen-primed, ovariectomized rats was adrenalectomized and tested for sexual receptivity following hypothalamic implantations of PGE2. The combination of PGE2 and adrenalectomy led to severe debilitation as manifested by greatly reduced open-field activity scores and inhibition of estrogen and progesterone induced sexual receptivity. Neither exogenous progesterone nor corticosterone was able to restore these behaviors to normal levels. A mechanism involving PGE2 and adrenalectomy-induced transient ischemia was discussed as a possible cause of the debilitation.


(-)-Hydroxycitrate and conditioned aversions.

The capacity of various salts of (-)-hydroxycitrate to produce conditioned rejection of a 0.25% saccharin solution was evaluated. The ethylenediamine salt of (-)-hydroxycitrate produced strong conditioned rejection of saccharin under both deprivation and nondeprivation conditions, but this effect was less than produced by equimolar doses of lithium chloride. The sodium salt of hydroxycitrate produced no conditioned rejection of saccharin in water deprived rats but did so in nondeprived animals. In these experiments, food intake was reduced by (-)-hydroxycitrate only during the first hour following administration of the drug. The magnitude of appetite rejection did not correspond to the degree of conditioned rejection, lending support to the conclusion that the food intake reduction was not merely a consequence of aversive effects of the drug.


The aftercare clinic as a residency training site. 'Community neuropsychiatry'.

Refocusing service and research priorities in psychiatry toward problems of major mental illness and its long-term management requires that more specific attention be paid to this area by residency training programs. A "community neuropsychiatry" residency training experience based in a state hospital aftercare clinic is described as one model for achieving this objective. Supplemented by didactic course work in mental health administration, the history and theory of community psychiatry, and experiences in other, varied service settings, this program has proved to be valuable for learning and teaching skills basic to community psychiatry practice.


[Dangerous side effects of lithium therapy].

A report is made on two cases of a pharmacogenic delirium as a result of combined lithium-haloperidol therapy, and on one case of lithium-induced diabetes insipidus renales with second-degree high pressure. Also, the development of a hydrocephalus internus male-resorptivus through lithium is discussed. In all patients there was a possible disposition due to old age or cerebral injury. Based on their effect on synaptic transmission the potentiating effect of lithium and haloperidol is discussed. Under the influence of Na-K stimulated ATPase and, thus, of active transport and adenylcyclase a basic mechanism of lithium therapy and its side-effects is seen.


[Islet cell adenoma with acute neuropsychiatric symptoms].

The author, by reference to observations made on two cases of islet cell adenoma with cerebral symptoms, describes the most important symptoms of this multifaceted disease picture. Diagnostic possibilities are discussed, and surgical treatment as the method of choice is pointed out.


[Pathomechanisms of myasthenic crisis states].

From the point of view of the kind of fate of those people ill with myasthenia the crisis itself can be regarded as the most dangerous and usually the most problematic phenomenon. To the crisis belong all those conditions characteristic of serious respiratory failure and those occurring either as a direct result of myasthenia or due to other causes associated with other illnesses. Clinically, there are three main kinds of crisis: myasthenic, cholinergic, and oscillatory/,brittle". There is the possibility of 3 directions of study on the crisis: The careful analysis, coordination, and observation of the clinical picture, with precise notation of the course of the illness. Analysis of the neuro-muscular types of blockade which occur in myasthenia. Interpretation of changes such as resistance and intolerance observed during the process and which conform to the pathomechanism. The author deals with problems mentioned with regard to his casuistry of 300 people ill with myasthenia.


[Studies of a full night's sleep in patients with absences].

Clinical electro-encephalography and especially the simultaneous recording of EEG values and other physiological parameters can greatly contribute to investigations into the neurophysiological aspects of sleep disturbances in patients suffering from attacks. In the case of absences peak potentials have been found in all phases of sleep, with a reduction in the pattern-like arrangement of the peak potentials and an increase in the irregular spike-and-wave complexes. During the dream phase, however, the peak potentials of these patients were primarily in the form of "patterns" comparable to the situation when they were awake. This evidence of epileptic activity throughout all sleep phases of patients suffering from absences makes it necessary to give anticonvulsive drugs in three doses distributed evenly over the entire day.


Pyruvate kinase isozymes of Mucor racemosus: control of synthesis by glucose.

A variety of cultural conditions were examined to determine the relationship between pyruvate kinase isozyme patterns and morphology in Mucor racemosus. The results indicate that M. racemosus has two isozymes of pyruvate kinase, form A and form B, which are clearly separable on ion-exchange columns (diethylaminoethyl-cellulose). Addition of glucose to cultures growing on amino acids in air resulted in the induction of form A and the termination of form B synthesis. Cycloheximide added at the same time as glucose blocked the formation of form A but did not interfere with the termination of form B synthesis. Removal of glucose resulted in termination of form A synthesis and the induction of form B. Cycloheximide blocked the induction of form B and did not interfere with the termination of form A synthesis. The data show that the isozyme type is not directly related to morphology, but depends only on the presence or absence of glucose.


Functional stability of the bfe and tonB gene products in Escherichia coli.

The expression of several functional properties of the products of the bfe and tonB genes in Escherichia coli was measured after the specific termination of the synthesis of the products of these genes. This was accomplished by the use of a temperature-sensitive amber suppressor mutation, which allowed control, by manipulation of the growth temperature, of the level of product formed from suppressible mutant alleles of the bfe or tonB gene. The bfe product is an outer membrane receptor protein for vitamin B12, the E-colicins, and bacteriophage BF23. The identity of the tonB product is unknown, but it is necessary for a subsequent step of uptake of vitamin B12, iron chelates, all of the group B colicins, and bacteriophages T1 and phi 80. Results from a different experimental system had shown that the termination of expression of the bfe locus was rapidly followed by loss of sensitivity to colicins E2 and E3 and, subsequently, to bacteriophage BF23. This was confirmed with this experimental system. Receptors that were no longer functional for colicin or phage uptake remained fully effective for B12 uptake, showing that receptors are stable on the cell surface. This supports previous contentions for the presence of different functional states for colicin receptors. The functional properties of the tonB product, measured by B12 uptake or sensitivity to the group B colicin D, were unstable, declining extensively after cessation of its synthesis.


Physiological function of superoxide dismutase in glucose-limited chemostat cultures of Escherichia coli.

Conditions for continuous culture of Escherichia coli K-12 His- Thi- under glucose limitation were established. Both the capacity for respiration, at D greater than 0.2/h, and specific activity of superoxide dismutase increased as a function of specific growth rate, whereas peroxidase and catalase were either invariant with or inversely related to this growth rate. The abrupt increase in the availability of glucose, as a means of elevating the growth rate, was followed by an increase in superoxide dismutase, which reached a plateau before there was a significant increase in the growth rate. Thus, an increase in superoxide dismutase appeared to be a prerequisite for an increase in the rate of growth. Cells that had higher levels of superoxide dismutase, because of varying specific growth rates, were more resistant to the toxicity of hyperbaric oxygen. Superoxide dismutase thus behaved like an essential defense against the toxicity of oxygen. Sensitivity towards streptonigrin increased with specific growth rate in the range of 0.09 to 0.25/h but decreased with further increases in the growth rate. Since this antibiotic has been shown to shunt electrons to oxygen, with concomitant production of O2-, these results indicated a progressive deficiency of reducing power at growth rates below 0.25/h and a surfeit of reducing power with progressively greater protection against O2- by superoxide dismutase at growth rates greater than 0.25/h.


Rates of ion movement from plasma to endolymph in the dogfish.

The rates of movement of Na+, Rb+, Cl- and HCO3- from plasma to endolymph were studied in the elasmobranch fish, Squalus acanthias, by use of the appropriate isotopes. Rb+ was used as a marker for K+. The half-times to equilibrium for Na+, Rb+ and Cl- were about 100 hours; for HCO3- it was 6 hours. The equilibrium ratios, endolymph/plasma, are Na+ 0.87, K+ 26, Cl- 1.37, HCO3- 1.47. Carbonic anhydrase inhibition decreased the rate of HCO3- accumulation, suggesting that the process is actually the formation of endolymphatic HCO3- from plasma or tissue CO2. Increase in plasma pCO2 elevates endolymph HCO3- concentration. The secretory tissue contains carbonic anhydrase and Na-K-ATPase. These and other data suggest that a dominant feature of endolymph chemistry may be HCO3- formation linked in some fashion with K+ transport, through rates catalyzed by these two enzymes.


Control of beta-glucosidase synthesis in Mucor racemosus.

The beta-glucosidase of Mucor racemosus was shown to be synthesized when the organism was grown in the presence of such diverse carbon sources as glycerol, lactate, xylose, ribose, alpha-methylglucoside, alpha-phenylglucoside, maltose, and cellobiose. Enzyme synthesis was strongly repressed in the presence of hexoses. In addition, exogenous cyclic adenosine 3',5'-monophosphate (cAMP) resulted in enzyme repression. When cAMP was added exogenously after enzyme activity had accumulated, a reversible enzyme inactivation occurred. Growth on disaccharides (maltose or cellobiose) was severely retarded in the presence of cAMP, whereas that on glucose remained unaffected. The results indicate a probable role for cAMP in control of glucosidase synthesis in Mucor.


Glycine metabolism by Pseudomonas aeruginosa: hydrogen cyanide biosynthesis.

Hydrogen cyanide (HCN) production by Pseudomonas aeruginosa in a synthetic medium is stimulated by the presence of glycine. Methionine enhances this stimulation but will not substitute for glycine as a stimulator of cyanogenesis. Threonine and phenylalanine are effective substitutes for glycine in the stimulation of HCN production. Glycine, threonine, and serine are good radioisotope precursors of HCN, but methionine and phenylalanine are not. Cell extracts of P. aeruginosa convert [14C]threonine to [14C]glycine. H14CN is produced with low dilution of label from either [1-14C]glycine or [2-14C]glycine, indicating a randomization of label either in the primary or secondary metabolism of glycine. When whole cells were fed [1,2-14C]glycine, cyanide and bicarbonate were the only radioactive extracellular products observed.


Changes in protein synthesis on mitomycin C induction of wild-type and mutant CloDF13 plasmids.

Mitomycin C treatment of Escherichia coli K-12 cells containing the nonconjugative plasmid CloDF13 resulted in inhibition of host chromosome protein synthesis and a high rate of synthesis of two CloDF13-specified proteins whose molecular weights correspond to cloacin and immunity protein. Five molecules of immunity protein were synthesized for each cloacin DF13 molecule. Mitomycin C-treated cells containing a copy mutant of CloDF13 made three to four times as much of each protein as cells containing wild-type CloDF13. CloDF13 plasmids that contained the transposon Tn1 were isolated. Two did not induce after mitomycin C treatment, failing both to inhibit host cell synthesis and to produce the two new proteins. In minicells, they showed reduced CloDF13-specified protein synthesis and produced three Tn1-specified proteins.


Analysis of Euglena gracilis chloroplast deoxyribonucleic acid with a restriction endonuclease, EcoRI.

Cleavage of chloroplast deoxyribonucleic acid (DNA) of Euglena gracilis Z with restriction endonuclease RI from Escherichia coli (EcoRI) yielded 23 bands upon electrophoresis in gels of agarose. Four of the bands contained twice the stoichiometric amount of DNA. One of these bands contained two similarly sized fragments. The sum of the molecular weight of the 24 different fragments equaled the molecular weight of the circular molecule. The restriction fragments had different buoyant densities, with four having distinctly heavy densities in CsCl. Restriction fragments with a high buoyant density were preferentially lost when broken chloroplast DNA was purified by equilibrium density gradient centrifugation. Hybridization of chloroplast ribosomal ribonucleic acid to intact chloroplast DNA determined that there are two cistrons for 16S and 23S ribosomal ribonucleic acid. These two cistrons are located on six restriction fragments, all of which have buoyant densities greater than the intact molecule of chloroplast DNA.


Characterization of lambda Escherichia coli hybrids carrying chemotaxis genes.

Molecular cloning techniques were used to construct hybrid Escherichia coli lambda phage and isolate Col E1 factors that carried the cheB region of the E. coli genome. The products of these genes were examined by using a series of deletions in the phage to stimulate specific polypeptide synthesis in ultraviolet-irradiated cells and by using Col factor to program protein synthesis in minicells. Seven flagellar related polypeptides were synthesized. Three of these with apparent molecular weights of 38,000, 28,000, and 8,000 were associated with the cheB region; three polypeptides 63,000, 61,000, and 60,000 were associated with the region that maps between cheB and cheA. These bands were referred to as the triplet group. We suggest that these polypeptides are the same as the methyl-accepting chemotaxis protein described by Kort et al. (Proc. Natl. Acad. Sci. U.S.A. 72:3939-3943, 1975). Another polypeptide with a molecular weight of 12,000 is associated with the cheA region which also produces at least three gene products. We conclude that the cheA-cheB region in E. coli is complex. Further genetic and biochemical analyses are required to describe all of these products.


Selected translocation of plasmid genes: frequency and regional specificity of translocation of the Tn3 element.

A procedure is described that selects for the insertion of transposable antibiotic resistance elements in a variety of recipient replicons. The selected translocation procedure, which employs a plasmid having a temperature-sensitive defect in replication as a donor of transposable genetic elements, was used to investigate certain characteristics of the translocation process. Our results indicate that translocation of the Tn3 element from plasmid to plasmid occurs at a 10(3)- to 10(4)-times-higher frequency than from plasmid to chromosome. In both cases, continued accumulation of Tn3 on recipient genomes is prevented by development of an apparent equilibrium when only a small fraction of molecules in the recipient population contain Tn3. An alternative method for estimation of translocation frequency has shown that the translocation process is temperature sensitive and that its frequency is unaffected by the presence of host recA mutation. Insertions of Tn3 onto the 65 X 10(6)-dalton R6-5 plasmid in Escherichia coli are clustered on EcoRI fragments 3 (8 of 23 insertions) and 9 (7 of 23 insertions), which contain 12 and 5%, respectively, of the R6-5 genome. The occurrence of multiple insertions of Tn3 within EcoRI fragment 9, which contains the IS1 element and a terminus of the Tn4 element, is consistent with earlier evidence indicating that terminal deoxyribonucleic acid sequences of already present transposable elements may provide recognition sequences for subsequent illegitimate recombinational events.


Dominance of dnaA+ to dnaA in Escherichia coli.

The dominance of dnaA+ to the dnaA508 mutation was complete and was unaffected by the presence of a copy of the chromosomal replication origin on the episome. These results prove that the dnaA gene of Escherichia coli produces a diffusible product.


Pelvic osteomyelitis in children.

Six children (7 to 16 years of age) with pelvic osteomyelitis are described. Sites of involvement included the pubis in three patients, the ilium in two patients, and the ischium in one patient. All were right-sided. Each patient presented with a history of fever and an abnormal gait. In four, the point tenderness indicated the site of bony involvement. All patients had pain on abduction but free passive range of motion of the hip. Soft tissue swelling was present on admission pelvic roentgenograms in five patients. Intravenous pyelogram revealed deviation of the bladder toward the midline in each of four patients studied. Roentgenographic changes typical of osteomyelitis developed in four patients ten days to ten weeks after onset of symptoms. In four patients in whom an organism was identified, Staphylococcus aureus was isolated from blood and/or bone. All isolates were methicillin-sensitive and two were penicillin-sensitive. Purulent material was drained from three of the five patients who underwent surgical exploration of the pelvis. All patients received parenteral antistaphylococcal therapy for 3 to 5 1/2 weeks (mean, 4 weeks). Oral antibiotics were given to five patients for an additional 3 to 14 weeks. All patients recovered completely.


Antimicrobial activity of metronidazole in anaerobic bacteria.

The antimicrobial activity of metronidazole was investigated in anaerobic bacteria by use of time-viability studies. This antimicrobial agent has a rapid onset of bactericidal activity under proper reducing conditions. The bactericidal rates were not affected by inoculum size or nutritional requirements, nor by inhibition of growth and protein synthesis by chloramphenicol. Using supernatant fractions of actively growing cultures of susceptible organisms, we observed a disappearance of metronidazole and a loss of biological activity, but there was no significant change in preparations from resistant bacteria. The decrease in drug concentration with susceptible cells occurred during the time that its bactericidal action was being exerted. Extracts from susceptible organisms rapidly reduced the concentration of metronidazole, confirming previous observations which suggest that the drug acts as a terminal electron acceptor. Radioisotope experiments with [14C]metronidazole revealed that the compound was taken up by both resistant and susceptible bacteria, although there was a difference in rate and extent of accumulation. These studies demonstrate that metronidazole's antimicrobial activity against anaerobic bacteria is bactericidal and independent of growth rate, and that it involves the uptake and metabolism of the compound.


5-Propyl-2'-deoxyuridine: a specific anti-herpes agent.

In both primary rabbit kidney cells and human skin fibroblasts, 5-propyl-2'-deoxyuridine proved inhibitory to herpes simplex virus at a concentration as low as 1 micrograms/ml, whereas concentrations higher than 200 micrograms/ml were required to inhibit vaccinia virus replication or normal cell metabolism.


Sensory conduction studies in the early recognition of nerve disorders.

Early investigations into sensory conduction are reviewed and related to recent and ongoing research, specifically with relation to the early detection of nerve disorders. Current study on entrapment neuropathies and on the assessment of minor sensory conduction changes is presented. It is concluded that, in the early recognition of nerve disease, the clinical orientation of the electromyographer is essential to proper test interpretation.


Early recognition of nerve disorders by near-nerve recording of sensory action potentials.

Evaluated herein are the advantages and limitations of recording sensory action potentials with a gross electrode placed near the nerve. This technique has proved to be a sensitive measure of early nerve damage for two reasons: first, electrodes can be positioned in such a way that maximal velocity is determined only along the abnormal stretch of the nerve; and second, small late components originating from demyelinated, remyelinated, or regenerating fibers can be detected.


Histology and ultrastructure of alterations in neuropathy.

Histologic findings are described in nerves from men exposed to lead, from patients with discrete clinical signs of peripheral neuropathy, and from controls. Every nerve from control subjects showed an abnormality (paranodal remyelination, segmental remyelination, or regeneration) in teased fibers. The only histologic alteration in eight lead-exposed males without signs or symptoms of neuropathy was a slightly increased incidence of paranodal remyelination. Sixteen patients with discrete neurologic symptoms and signs had a loss of large myelinated fibers and an increased incidence of regenerated fibers among teased fibers. Electron microscopy of unmyelinated fibers showed an increased occurrence of Schwann-cell processes, of fibers undergoing degeneration, and of Schwann-cell subunits with many profiles as the earliest signs of abnormality. Clinically mild neuropathies may exhibit advanced regeneration in the case of unmyelinated fibers. The earliest sign of degeneration in myelinated fibers was a diminution in the number of axonal organelles.


Recognition of unmyelinated fiber disease: morphologic criteria.

An attempt is made to evaluate available information regarding the integrity of unmyelinated fibers, particularly somatic unmyelinated fibers in man and in other primates, with relation to early recognition of nerve disorders. Discussion is limited to morphologic aspects of these fibers.


Neurophysiological investigation of subclinical and minimal toxic neuropathies.

In the investigation of subclinical or minimal toxic neuropathies, selection of the proper electrophysiological test depends largely on the nature of the condition being studied. Examples are given of several toxic disorders, and their features are discussed in relation to the appropriate electrophysiological measurement.


Screening for peripheral neuropathy in patients treated by chronic hemodialysis.

Observations were made on the occurrence of peripheral neuropathy in a series of 139 patients admitted to a chronic dialysis program over a 10-year period. Evidence of neuropathy was obtained in approximately 50% of these patients over the total period. Once dialysis was instituted, distal paresthesiae evident before the inception of dialysis tended to clear rapidly. Occurrence of the symptom of "restless legs" correlated positively with the presence of neuropathy. Persistent neuropathy was more commonly sensory than motor, and its features were consistent with a predominant loss of large myelinated fibers. This analysis suggests that multiple factors may be involved in the origin of uremic neuropathy.


Magnetic resonance studies of Mn(II)-, Mn(III)-, and Fe(III)-conalbumin complexes.

Apoconalbumin binds Mn(II) at two sites with association constants of K1 = 7 (+/- 1) X 10(4) and K2 = 0.4 (+/- 0.25) X 10(4) M-1. The binding is tighter in the presence of excess bicarbonate resulting in K1 = 1.8 (+/- 0.2) X 10(5) and K2 = 3 (+/- 2) X 10(4) M-1. The electron paramagnetic resonance spectrum (at both 9 and 35 GHz) of Mn(II) bound at the tight site reveals a rhombic distortion (lambda = E/D approximately equal to 0.25-0.31) in the protein ligand environment of the mental ion. An evaluation of the 1/pT1p, paramagnetic contribution to the longitudinal relaxation rate of solvent protons with Mn(II)-, Mn(III)-, and Fe(III)-derivatives of conalbumin revealed that the mental ion in each site of conalbumin is accessible to one water molecule. For Mn(II)-conalbumin and Mn(III)-conalbumin species, inner coordination sphere protons are rapidly exchanging with the bulk solvent, while slow exchange conditions prevail for Fe(III)-conalbumin.


The effect of anticholinesterases on motor unit potentials in myasthenia gravis.

In most patients with myasthenia gravis, the mean duration and amplitude of motor unit potentials sampled in the extensor digitorum communis muscle, as well as the amplitude of the pattern of activity during full effort, increased after application of anticholinesterase. This appears to be a sensitive test for early detection of failure in neuromuscular transmission. In a series of 12 patients, the mean amplitude of motor unit potentials increased 30%-130% in 9, and the mean duration increased 10%-25% in 7. In 3 patients who failed to show an increase in amplitude in the extensor digitorum communis muscle, the amplitude of motor unit potentials increased in a facial or extraocular muscle after intravenous application of edrophonium. This increase in the amplitude or duration of motor unit potentials was even evident in 4 patients who failed to show a decrement in response of the opponens pollicis muscle to repetitive stimuli delivered to the median nerve at 3/sec and 5/sec. In 5 patients, responses evoked by stimuli delivered at 15/sec showed marked facilitation.


Secretory endometrium associated with breakthrough bleeding in estrogen-treated postmenopausal women.

Paradoxical secretory changes are described in benign endometria curetted for breakthrough bleeding in postmenopausal women on cyclic low dosage Premarin therapy. This purely histologic study, though favoring progesterone mediation, provides no definitive answer to the problem of pathogenesis. Future correlative morphologic and endocrine investigations are clearly indicated.


Incidental pregnancy luteoma associated with ectopic tubo-ovarian pregnancy: evidence in favor of origin from theca interna of an atretic follicle.

An incidental pregnancy luteoma was identified microscopically in an ovary submitted for ruptured ectopic tubo-ovarian pregnancy. The lesion consisted of a localized, non-encapsulated proliferation of thecal cells in the wall of an atretic follicle and had all the pathologic features of the typical late pregnancy luteoma. This finding is presented to support the origin of the pregnancy luteoma from the theca interna of atretic follicles.


Immunoprophylactic anti-Rho(D) treatment after mismatched transfusions.

80 cases of anti-Rho(D) treatment after mismatched transfusions are reviewed to compare the intramuscular (i.m.) and intravenous (i.v.) route of treatment. No severe reactions such as renal failure occurred with either method of anti-D treatment. If the Rh-immune globulin is injected i.m., 20 micrograms anti-D per ml red cells are used. For i.v. anti-D administration, 10-12 micrograms anti-D are suggested. The i.v. method of anti-D treatment is recommended for the future. Furthermore, a scheme of treatment for after mismatched transfusions is described.


Diffusion rates of cell surface antigens of mouse-human heterokaryons. I. Analysis of the population.

The rate of appearance, in a newly formed heterokaryon population, of cells bearing completely intermixed mouse and human surface antigens may be used to estimate diffusion constants for antigens on individual cells. From this estimate, it appears that the surface antigens in most cells do not diffuse at the rate expected, but rather move more slowly, by a factor of ten or more, than expected from either measured or calculated diffusion constants for proteins freely mobile in the plane of a lipid membrane. Differences in diffusion rates between cells are not due to effects of Sendai virus, or of trypsin. Restrictions on diffusion are apparently not due to cytochalasin B- or Colcemid-sensitive elements.


Topographical localization of the receptors for luteinizing hormone-releasing hormone on the surface of dissociated pituitary cells.

A derivative of the hypothalamic peptide luteinizing hormone-releasing hormone (LHRH) has been coupled to ferritin and the conjugate purified by gel chromatography. In its ability to stimulate the secretion of luteinizing hormone from pituitary cells in vitro, the conjugate has the same potency and specificity as the native peptide. When dissociated pituitary cells maintained in short-term culture are lightly fixed with formaldehyde and then incubated with the conjugate, examination in the electron microscope shows an even distribution of ferritin particles over the free cell surface of the gonadotrophin cells. This binding appears to be specific for the LHRH receptor since it is prevented by a 10-fold excess of native peptide. In addition to the gonadotrophin cells, some somatotrophin and thyrotrophin cells bind conjugate on their free surfaces under similar conditions. If living cells are incubated with the conjugate for 15 min, the bound conjugate becomes aggregated and then concentrated in one localized area of the cell surface. In this area, which lies immediately above the juxtanuclear Golgi complex, the plasma membrane is frequently invaginated in a manner which suggests that the bound, aggregated conjugate is internalized by endocytosis.


Juvenile nasopharyngeal angiofibroma: management of intracranial extension.

Juvenile nasopharyngeal angiofibroma (JNA) is a histologically benign, locally invasive tumor of the nasopharynx that is found primarily in the pubescent male. While most authors recommend surgical excision for smaller, localized extracranial tumors, opinion varies on the management of more aggressive tumors with intracranial extension. The authors present their experience over the past 15 years with 15 cases of JNA with intracranial extension, and formulate a plan of management. A combined otolaryngologic/neurosurgical approach is recommended in order to assess the extent of the tumor and to isolate feeding vessels. Lateral extension into the middle cranial fossae should be resected in continuity with the nasopharyngeal component, which is approached from below in a separate operative field. Medial extensions are transected, verified angiographically three months after surgery, and irradiated. The results of therapy are presented, and a case of dural penetration by JNA is described.


Is there a role for actin in virus budding?

Electrophoretic data from both sodium dodecyl sulfate-polyacrylamide gels (SDS-PAGE) and acid-urea gels reveal a protein in purified murine mammary tumor virus (MuMTV) which co-migrates with purified chick skeletal muscle actin. 125I-labeling of intact and disrupted virus preparations shows that the actin-like protein is not artifactually adsorbed to the outside of virions during isolation. Quantitative SDS-PAGE and examination of negatively stained preparations show that the actin cannot be accounted for by a contaminating population of virus-free vesicles. The ultrastructure of mammary epithelial cells and of Rous sarcoma virus-transformed chick embryo fibroblasts shows that virus extrusion is associated with filament-containing cellular processes. In particular, MuMTV is released from the ends of long microvilli which contain a bundle of 6-8-nm microfilaments and share other structural features with intestinal microvilli. We suggest that virus nucleoids require an interaction with host cell contractile proteins for their extrusion from the cell.


Glomus tumour of the fingers.

Clinical and therapeutic aspects of glomus tumour of the fingers have been considered by analyzing the records of 15 patients with glomus tumours. The tumour was found in the pulp in 5 patients and in the subungual region in 10. The diagnosis may be difficult to make and patients are often believed to be malingering. Complete surgical excision under magnification is curative.


Labile (borderline) hypertension--new aspects of a common disorder.

Labile hypertension in patients under 50 years of age (the non-atherosclerotic form) was found to be characterized by higher urinary excretion of catecholamines and particularly of homovanillic acid; when further analyzed it was shown to be a heterogeneous entity with two types of patients clinically and biochemically distinguishable from each other, from control subjects and from patients with stable hypertension. Reactivity to assuming an upright posture distinguishes one type of labile hypertension having a normal postural pulse rate response from another having an excessive postural increase in pulse rate. The first group also showed normal responses of plasma norepinephrine concentration and of urinary cyclic AMP to posture. The group with excessive pulse rate response, in contrast, showed a decrease in plasma norepinephrine and an excessive increase of urinary cyclic AMP excretion in response to upright posture. The results suggest that not only circulating catecholamines but also the reactivity of their target tissues (as probably reflected by cyclic AMP measurements) are important in bringing about signs of adrenergic excess. The hypothesis that cyclic AMP changes reflect beta-adrenergic receptor reactivity is strongly favoured by data indicating qualitative differences in cyclic AMP responses to beta-adrenergic stimulation or inhibition between control subjects and those labile hypertensive patients with clinical signs of excessive sympathetic reactivity. The study stresses the need for more precise definition of labile hypertension, for dynamic clinical and biochemical correlative studies, and for consideration not only of the circulating hormones but also of the "second messengers" (such as cyclic AMP and cyclic GMP) which reflect the cellular action of hormones. Blood pressure is a very labile parameter in health and disease. In one sense, therefore, hypertension can be considered "labile" in every hypertensive patient. Usually, however, labile (or borderline) hypertension is regarded as characterized by a blood pressure over 140/90 mmHg, falling below these values with physical and emotional rest. This clinical entity, which affects some 20% (variously estimated between 16 and 30%) of the adult population, gives rise to uncertainties in both definition and prognosis. In some patients labile hypertension represents the precursor of a fixed hypertensive state, whereas in many others it remains labile throughout life, never progressing to the stable phase nor becoming associated with hypertensive cardiovascular disease.(ABSTRACT TRUNCATED AT 400 WORDS)


Fibrillar bodies in leukemic cells revealed by polarization microscopy.

An optical polarizing microscope with a good coefficient of extinction permits the visualization of the cytoplasmic fibrillar body in living preparations and smears of leukemic cells (human leukemias and the L 5222 experimental leukemia). These inclusions are not visible by phase contrast microscopy nor in fixed and stained smears. The detection in living cells of fibrillar bodies makes it possible to study directly the conditions for their formation and their reaction to the effect of certain drugs.


[Role of thermography in the diagnosis of breast neoplasms].

319 cases of breast pathology of doubtful clinical diagnosis were studied by means of thermography and mammography, with histological verification: 214 proved to be malignant tumours and 105 benign. The role of thermography is discussed on the basis of these results; it would appear to be restricted to diagnostic aid status associated with other procedures, particularly clinical examination and mammography.


[Intestinal transit time in Togo (Western Africa) and Germany].

In five groups of Africans and Europeans in Togo (West-Africa) and Germany intestinal transit time and stool-weights have been studied. Native Africans on traditional food passed three times more stool (307 g/d) in half of the time (33 hours) as Europeans and Africans on Western diet did. The effect of dietary fibre on intestinal transit time and intestinal pathology is stressed.


Screening for neurobehavioral toxicity: the need for and examples of validation of testing procedures.

The need for a sensitive and reliable screen to assess environmental agents for potential behavioral and neurological toxicity is discussed. Factors involving strategy, choice of animals and doses, route of administration, duration of study and requirements for the selection of neurobehavioral tests are also evaluated. The primary emphasis concerns the need for standardization and validation of neurobehavioral tests to be used in neurotoxicology. It is suggested that test validation be accomplished by comparing the observed results of known neurotoxicants in animal models which are chosen to predict effects based on reported human symptomatology. As a means of demonstrating how test validation is used in our laboratory, data from a number of experiments concerning the effects of a variety of chemical agents on three measures of motor functioning were discussed. The neurobehavioral effects of acrylamide, and agent known to produce "dying-back" axonopathies, were assessed using separate techniques presumed to measure hindlimb and forelimb functioning and general motor activity. The prediction that acrylamide will first decrease hindlimb functioning, while decreasing forelimb grip strength and motor activity at higher doses, was confirmed. The validity of the hindlimb measurement was supported using a neurotoxicant, carbon disulfide, known to affect motor functioning in a manner similar to acrylamide. The validity of the forelimb technique was shown indirectly using normative data collected from rats of both sexes tested at various ages, i.e., males were stronger than females and grip scores changed as a function of age. The relative sensitivities of the fore- and hindlimb measurements were found to be approximately the same when used to assess the effects of known muscle relaxants, such as phenobarbital and chlordiazepoxide. Finally, it was predicted and confirmed that an environmental agent believed to affect behavior secondarily to effects on other organ systems would affect all measures of motor functioning at approximately the same dose.


Use of discrimination behavior for the evaluation of toxicants.

This study involved the application of discrimination behavior for the study of effects of environmental contaminants on the behavior of laboratory animals. Polybrominated biphenyl (PBB) was evaluated for effects on the acquisition and performance of a simple auditory discrimination by rats. Methyl ethyl ketone (MEK), methyl isobutyl ketone (MIBK) and carbon monoxide (CO) were evaluated for effects on a delayed match-to-sample discrimination task in the juvenile baboon. All of the contaminants slowed response times and increased extra responses. These findings suggest that discrimination behavior may be of value for the evaluation of environmental contaminants for effects on the central nervous system.


Insoluble "fibrin" in human aortic intima. Quantitative studies on the relationship between insoluble "fibrin", soluble fibrinogen and low density lipoprotein.

A quantitative assay for fibrin or other insoluble fibrin-like antigens ("fibrin") in small samples of intima is described. Tissue samples were subjected to electrophoresis directly from the intima into an antibody-containing gel to remove and measure fibrinogen and other soluble fibrin reactive antigens (FRA). The residual tissue was then exhaustively incubated with plasmin, and the soluble fragments generated from the insoluble "fibrin" were measured by quantitative immunoelectrophoresis. "Fibrin" accounted for about 2% of the tissue dry weight in normal intima and the ratio fibrinogen/"fibrin" was 1-1.5. In the gelatinous lesions, which seem to be the precursors of fibrous plaques, there was a small increase in "fibrin" but a substantial increase in fibrinogen and low density (LD)-lipoprotein, and the ratio fibrinogen/"fibrin" rose to about 3, which suggests that the increase in "fibrin" is secondary to increased permeation of fibrinogen. At the edges of large plaques there was also a threefold increase in fibrinogen, but "fibrin" increased fivefold, and accounted for 10% of the tissue dry weight. The same high concentration was found in the centres of large fibrous plaques with advanced atheroma lipid. Raised levels of "fibrin" were accompanied by raised levels of fibrinogen in most tissue samples. About 80% of the total soluble FRA could be clotted with thrombin; there was no significant difference between normal intima and lesions, and the proportion clotted was not related to "fibrin" content.


Participation of calcium in flagellar shortening and regeneration in Chlamydomonas reinhardii.

Cation chelators cause flagellar shortening in Chlamydomonas reinhardii. Most effective are EDTA and EGTA (1 mM) but pyrophosphate (10 mM) also is effective. Addition of 5 mM Ca2+ after shortening caused by 4 mM EGTA results in flagellar regeneration. Other divalent cations can replace Ca2+ with the following relative activities: Ca2+ greater than Sr2+ = Mn2+ much greater than Ba2+ = Mg2+. Although the specific ion requirement to reverse shortening is not clear, it is possible that all of the ions act by displacing one bound cation, presumably Ca2+. A specific requirement for Ca2+ in flagellar regeneration could be demonstrated, however, because as little as 50 microM EGTA in the presence of 500 microM Mg2+ delayed regeneration and prevented full regeneration. Ca2+ at 100 microM overcame this inhibition.


Very low density lipoproteins in normal and cholesterol-fed rabbits: lipid and protein composition and metabolism. Part 1. Chemical composition of very low density lipoproteins in rabbits.

The lipid and apoprotein composition of very low density lipoproteins (VLDL) (d less than 1.019 g/ml) from normal and hypercholesteremic (h.c.) rabbits are compared. Significant changes are observed in the apoproteins, as well as in the fatty acid composition of cholesterol esters, and in the relative distribution of phospholipids. Apoproteins show a marked increase of peptides with slow mobility (R2 and R3), corresponding to arginine-rich proteins, both in the rabbit and in some types of human hyperlipoproteinemias (hypothyroidism and Fredrickson Type III). VLDL were separated into the two fractions, VLDL-1 and VLDL-2. Peptides of slow mobility were shown to be present in a higher concentration in VLDL-1. Cholesterol esters of h.c. VLDL have a very high content of oleic acid, the 18:1/18:2 ratio being about three times higher than in normal VLDL. Phospholipid distribution in h.c. VLDL is characterized by an increase of sphingomyelin and decrease of phosphatidylethanolamine. The PC/Sph ratio is about one fifth that of normal. These changes in VLDL lipids are very similar to those occurring in the lipid composition of the atherosclerotic plaques in mammals.


Cellular recognition by rat liver cells of neuraminidase-treated erythrocytes. Demonstration and analysis of cell contacts.

Freshly isolated rat liver cells adhere firmly to neuraminidase-treated rat or mouse erythrocytes but not to untreated erythrocytes. Binding between cells occurs only in the presence of calcium and is specially inhibited by D-galactose. We therefore suggest that cell adherence is mediated by a galactose-specific hepatic membrane receptor. Ultrastructural analysis of contact regions revealed point-like interactions between hepatic microvilli and erythrocytes and no broad areas of membrane contact. When liver cells are cultivated in vitro they lose their ability to bind erythrocytes within 24 h.


Very low density lipoproteins in normal and cholesterol-fed rabbits: lipid and protein composition and metabolism. Part 2. Metabolism of very low density lipoproteins in rabbits.

The metabolic fate of very low density lipoproteins (VLDL) in normal and hypercholesteremic (h.c.) rabbits has been investigated. VLDL were labelled with 125I in the protein moieties and injected into normal and h.c. animals. The turnover of h.c. VLDL is markedly delayed as compared to that of normal VLDL, and conversion into lipoprotein classes of higher density is considerably decreased. This is observed when h.c. VLDL are injected either into h.c., or into normal rabbits. Arterial uptake of radioactivity is much higher with h.c. VLDL than with the normal lipoproteins, and it is highest when h.c. VLDL are injected into normal recipients. These data, together with those reported in the previous study, support the hypothesis that h.c. VLDL have an inherent atherogenicity. Injection of h.c. VLDL into normal animals also offers an experimental model for testing drugs or diets against atherosclerosis, using untreated animals.


Nuclear control of neurite induction in neuroblastoma cells.

Induction of neurite formation in neuroblastoma cells by dibutyryl cyclic 3':5'-AMP (db-cAMP) or prostaglandin EI (PGE1) was enhanced after enucleation. Cells selected for resistance to db-cAMP were induced to form neurites by db-cAMP or PGE1 only after, but not before enucleation. Inhibition of protein synthesis inhibited neurite induction in nucleated, but not in enucleated cells, and enucleated cells were less sensitive to inhibition of neurite formation by concanavalin A (ConA). Colchicine, vinblastine and cytochalasin B (CB), compounds that interfere with the assembly of microtubules and microfilaments, inhibited induction in both types of cells. It is suggested that enucleation removes a nuclear inhibitor of neurite induction by db-cAMP and PGE1, and that neurite induction in nucleated cells requires that cAMP activates the assembly of microtubules and microfilaments and inactivates the nuclear inhibitor.


Aggregation of sponge cells. XIV. Possible substitution of calcium ions by polycations.

Single cells from the siliceous sponge Geodia cydonium, obtained after chemical dissociation, reaggregate in the presence of the aggregation factor in Ca2+-containing medium to large aggregates. It was found that polyvalent organic cations (polylysine, spermine, spermidine, putrescine) enhance the Ca2+-mediated cell aggregation. In Ca2+-free medium these compounds also cause reaggregation; aggregates of a diameter up to 800 micron are formed within 120 min. Proteins, containing basic groups of amino acid residues have no influence on cell aggregation. Monovalent cations inhibit the reaggregation process. The enhancing effect of polyvalent organic cations on cell aggregation is dependent on the presence of the soluble aggregation factor. From the findings that polycations do not alter the duration of the lag phase (a characteristic of the aggregation factor-mediated Geodia cell reaggregation) and act in cooperation with the aggregation receptor, we assume that the polycations bind between the aggregation factor and the aggregation receptor.


Sister chromatid exchange in patients with viral disease.

Sister chromatid exchange (SCE) frequencies were studied in differentially stained chromosomes from lymphocytes of 17 patients with viral disease. The mean SCE score for the patients was 8.7 +/- 2.9 standard deviations. SCE scores were significantly elevated in the patients compared with the controls (p less than 0.01); however, variability in SCE means was observed in the patients. SCE elevations were also present in long term cultured Epstein Barr virus positive human B lymphocytes.


Tissue metabolism and enzyme activities in the rodent Heterocephalus glaber, a poor temperature regulator.

1. Tissue oxygen uptake and enzyme activities were investigated in the naked mole rat, Heterocephalus glaber, a mammal notable for its low body temperature and metabolism and poor temperature regulating ability. 2. Q10 for O2 uptake of Heterocephalus crude liver homogenates ranged from 1.91 for the temperature interval 25-30 degrees C to 1.76 within the range 30-38 degrees C, values similar to those reported for typical homoiotherms. 3. Km pyruvate of lactate dehydrogenase in heart muscle had the same temperature dependence in the mole rat and mouse. 4. O2 uptake and cytochrome oxidase activity of skeletal muscle were higher for mole rat than mouse. The reverse was true for heart muscle. Brain and liver O2 uptake showed similar values for both species, while kidney O2 uptake was highest in the mouse. 5. Pyruvate kinase activity in heart and skeletal muscle was higher in mouse than mole rat, suggesting a greater reliance on glycolysis in the former. 6. Na+, K+ -ATPase activity of liver and kidney was 60% higher in mouse than mole rat, while brain was 30% higher in mouse. 7. The results indicate that the effects of temperature on tissue metabolism in the mole rat conform to those in typical homoiotherms. The low body temperature and O2 uptake in the mole rat find no expression in the tissue respiratory capacity.


The perchlorosoluble proteins of the serum of the rainbow trout (Salmo gairdnerii Richardson): albumin like and hemoglobin binding fraction.

1. The main perchlorosoluble fraction of rainbow trout serum has some physico-chemical characters kindred to those of human serum albumin (low molecular weight, solubility with ammonium sulfate, electrophoretic mobility, no glycoproteinic staining). 2. However, on account of obvious differences (heterogeneity and existence of various phenotypes, lack of bromophenol blue or bilirubin binding, low concentration, solubility in perchloric acid), the term "para-albumin" seems more suitable to name this compound. 3. The perchlorosoluble fraction binds hemoglobin causing an increase of peroxidasic activity. But, unlike to human haptoglobin, hemoglobin binding is partial, reversible and labile.


Cyclic nucleotides in nervous tissue.

A review of the literature emphasizes that cyclic nucleotides play a key role in regulation of cell growth, differentiation and metabolism in diverse tissues and, in addition, are closely involved in neural tissue function. The role of cAMP as a second messenger is discussed.


Variations in (Na+ + K+)-ATPase and Mg2+-ATPase activity of the ground squirrel brain during hibernation.

1. The specific activity of brain (Na+ + K+)-ATPase and Mg2+ -ATPase of the ground squirrel (Spermophilus richardsonii) is significantly increased after long-term hibernation. 2. The markedly non-linear thermal dependence of (Na+ + K+)-ATPase is unchanged during hibernation whereas the near linear thermal dependence of Mg2+-ATPase undergoes minor alteration after prolonged hibernation. 3. The sensitivity of (Na+ + K+)-ATPase to inhibition by ouabain is significantly decreased after 100 days of hibernation as is both the rate and amount of [3H]-ouabain binding. 4. These changes may be related to alteration in the phospholipid matrix of the membrane rather than alteration in the protein structure of the enzyme.


Serum lipoproteins in the spiny lobster, Panulirus interruptus.

1. Most of the lipids in the hemolymph of the spiny lobster, Panulirus interruptus, were associated with a high density lipoprotein (HDL3). The lipid of this lipoprotein was composed of phospholipid (88%), sterol (4%) and triglyceride (3%). 2. In animals fed 14C-labeled triglyceride radioactivity was not seen in the serum until 12 hr after feeding. Most of this serum radioactivity was associated with phosphatidyl choline. 3. Electron micrographs showed that negatively stained high density lipoproteins of the lobster had a polymorphic appearance.


Protein phosphorylation in human synaptosomal membranes: evidence for the presence of substrates for cyclic nucleotide guanosine 3'-5'-monophosphate dependent protein kinases.

Synaptosomal membranes prepared from different anatomical regions of postmortem human brain readily incorporate phosphate when incubated with labelled ATP in vitro. Separation of proteins on SDS slab gels indicated up to 30 protein bands stained by Coomassie blue of which ten incorporated label, as detected by radioautography (mol. wt. range 16-110 K with a major double band at 49-50 K). Incorporation into the 110 K region appeared to increase in older persons, and that into the 49-50 K region decreased. Human but not rat membranes subjected to similar conditions of treatment failed to respond to the addition of cAMP at 10(-5) - 10(-7) M but were highly sensitive to the addition of cGMP at 10(-6) M, which led to intensification of existing bands and the appearance of a major band at 60 K. The 110 K band present in high concentration in white matter striatum, caudate nucleus, and putamen was insensitive to cGMP addition. The presence of cGMP-dependent protein kinase substrates has not previously been reported in synapses, and it may be of significance in view of the known sensitivity of this system to biological regulatory agents.


Involvement of cholecalciferol metabolism in birds in the adaptation of calcium absorption to the needs during reproduction.

1. The metabolism of calcium and cholecalciferol in quail (Coturnix coturnix japonica) and chicken (Gallus domesticus) during maturation was correlated to gonadal activity and plasma oestrogen levels. 2. Birds with undeveloped ovaries (immature), developed ovaries but not laying (mature), and after laying 3-8 eggs (laying), were used in the first series. 3. Birds in which egg production had been arrested by Nicarbazin, were used in the second series. 4. Plasma 17 beta-oestradiol and calcium were elevated in the mature bird, with no further change in the laying bird. Kidney 25-hydroxycholecalciferol-1-hydroxylase and intestinal calcium-binding protein increased slightly in the mature bird, whereas they were grossly elevated in the laying bird. 5. Calcium and phosphorus absorption were markedly elevated in the laying bird. 6. No changes were noted in plasma 25-hydroxycholecalciferol, at any stage of maturation. 7. During the arrest of egg production by Nicarbazin, 17 beta-oestradiol level, calcium concentration of plasma, and medullary bone were maintained. Kidney 25-hydroxy-cholecalciferol-1-hydroxylase, intestinal calcium-binding protein and absorption of calcium were strikingly reduced. 8. The results suggest that changes in calcium absorption and cholecalciferol metabolism during maturation in birds are not directly affected by gonadal hormones; they appear to represent an adaptation to the increased calcium needs due to medullary bone formation and, more importantly, to the large losses of calcium imposed by shell formation.


A study of bovid herpesvirus 2 infection in calves inoculated intradermally or intranasally.

Bovid herpesvirus 2 infection was studied in calves exposed to the virus by intradermal inoculation of the skin of the left cheek or by nasal spray. In either case a localised infection developed and virus replication was shown to occur mostly in the tissues of its primary localisation, i.e. the skin of the left cheek or the nasal mucosa. There were neither secondary lesions, except at the site of virus injection, nor any serious systemic involvement on the part of the animals. The virus was also recovered from some areas of the skin (right cheek, perineum and scrotum) that were free of macroscopic lesions; moreover, intranuclear inclusions were found in several tissues of the nervous system (brain, superior cervical, stellate and Gasserian ganglia) which did not show any signs of inflammatory or degenerative changes. These findings suggest that the skin and the nervous system play an important role in the naturally-occurring disease since they could be the sites where the virus is maintained latently in the host.


A new freeze-dried living virus vaccine against sheep-pox.

A sheep-pox virus strain has been adapted and multiplied in primary lamb kidney cell cultures. The main characteristics of the strain have been verified in vitro after clones were isolated, and the results confirmed its identity. The safety and the potency of the strain have been investigated in sheep. The inoculation of the strain to sheep was followed by a post-vaccinal reaction materialised by a nodule at the site of inoculation and an increase of temperature by about 1 degree C. No reactions adversely affecting pregnancy have been noted. Immunisation was demonstrated by an increase in the level of neutralising serous antibodies and protection against the pathogenic virus. The immunity tended to decrease during the second year after primovaccination and a yearly booster vaccination appeared to be necessary. Primovaccination of lambs over 2 months of age produced a better immunity, especially when the lambs were born from vaccinated ewes. This strain forms the active principle of a freeze-dried vaccine containing no adjuvant of the immunity.


Immunisation against panleukopenia: early development of immunity.

The time necessary to obtain the immunity of cats against Panleukopenia has been studied by means of a modified live vaccine. This vaccine makes it possible to obtain a very early post-vaccinal immunity: the full immunity is reached 72 hr after the inoculation of the vaccine by the subcutaneous route. Furthermore, we have demonstrated that a sensitive kitten can be admitted in a contaminated environment immediately after vaccination without showing any clinical evidence of the disease.


Bovine adenoviruses--IV. Two mixed antigens for routine serodiagnosis by complement fixation reaction.

The frequency of bovine adenovirus infections occurring in cattle has been grossly underestimated up to the present day. Primary isolation of serotypes belonging to bovine subgroup II is cumbersome. Hardly any laboratory has used all 8 officially-recognised bovine serotypes in seroneutralisation-tests, which should be done when this type-specific method is used for serodiagnosis. The complement fixation test, known as group-reactive from human adenovirus serology, has failed to disclose the true incidence of bovine infections, as until recently the importance of a novel additional group-reactive bovine adenovirus antigen has been undisclosed. Here we describe production, composition and performance of two mixed antigens for complement fixation reactions, which take into account recent findings by our team on peculiarities of bovine adenovirus antigens. Mixed antigen 1 contains bovine serotypes 1, 2 and 3 and detects group-specific antibodies of the classical mastadenoviruses. Mixed antigen 2 contains bovine serotypes 4, 5, 6, 7 and 8 and determines group-specific antibodies against the novel paramastadenoviruses. In addition, each antigen is capable of demonstrating complement-fixing type-specific antibodies in sera against the respective types included in each mixed antigen, with a net enhancing effect produced by mixing. Use of the 2 mixed antigens promptly shows whether bovine adenoviruses, presently recognised as well as unclassified, are involved in a given outbreak of respiratory or enteric disease of cattle. If needed, the responsible type may be determined in a second step of serological investigation.