Immunological cross-reactivity between basic proteins of myelin and cancer. I. Lymphocyte transformation studies in immunized guinea-pigs.

The studies described were designed to examine the question of cross-reactivity between basic protein of myelin and a basic protein common to many human tumours. The lymphocytes from guinea-pigs injected with acid extracts of human cancer tissue showed significant (P less than 0-01) tranformation on exposure to basic protein of human myelin. Conversely lymphocytes from guinea-pigs injected with basic protein of human myelin showed significant (P less than 0-01) transformation on exposure to the acid extracts of human cancer tissue. Lymphocytes from control non-injected guinea-pigs did not transform on exposure to either antigen. These findings demonstrate immunological cross-reactivity in guinea-pigs between basic proteins of human myelin and of human cancer tissue using an assay system other than the macrophage electrophoretic migration hitherto used to show this effect in man.


Smooth-muscle antibodies and other tissue antibodies in cytomegalovirus infection.

Smooth-muscle antibodies (SMA) were present in 16% of sixty-three patients with cytomegalovirus (CMV) antibodies in serum and absent in forty CMV antibody-negative blood donors (P = 0-005). The SMA were of the IgG and IgM class, while no IgA antibodies were found. In patients with CMV infection, SMA, mainly of the IgM class, were present in the early stages of the disease, and the titre decreased faster than the complement-fixing CMV antibody titre. Anti-nuclear antibodies (ANA) were also found more often in CMV antibody-positive sera than in CMV antibody-negative sera, and ANA were usually present in sera which also contained SMA. Parietal cell antibodies, mitochondrial antibodies and other cytoplasmic antibodies did not occur more frequently in CMV antibody-positive than in CMV antibody-negative sera.


Viruses as teratogens.

There is nothing unique about rubella viruses or cytomegaloviruses. They are classic teratogens and, in this respect, not only follow but exemplify most of the principles of human teratology.


Endocrine assay in chorionic tumors.

Gestational chorionic tumors represent a rare biologic system for three reasons: They are a genetic mixture of both male and female; they are unusually responsive to chemotherapy with a high cure rate; and they always secrete hCG, which acts as a reliable cell marker with which to diagnose, to monitor drug therapy, and to follow-up to ensure continued remission. Nongestational chorionic tumors characteristically share only the latter unique attribute. For this reason, endocrine assays, particularly the measurement of hCG, play a vital role in the management of patients with these tumors. However, as the tools with which we work improve, and our understanding increases, it is quite likely that other tumors will be found to make substances that are similarly specific and reliable so as to be utilized in the same way. Already there are tests such as alpha-fetoprotein and carcinoembryonic antigen that are beginning to serve such a role but are not as specific as is hCG. Our experience with chorionic tumors has given us a valuable opportunity to develop techniques to be used for other tumor systems when reliable cell markers become available. When this time comes, we should be able to look forward to improved survival rates, hopefully approaching those obtained with chorionic disease.


Syndactyly of upper limb. Morphogenesis, classification, and management.

The distinction between primary and secondary syndactyly is made on the basis of their relation to the morphogenesis. The latter is associated with antecedent malformation which results in subsequent refusion of parts. Based on these considerations, a practical classification is suggested. The management of the various types of malformations is considered in relation to the complexity of the structures involved. Certain deformities require very early treatment in order to permit prehensile function to the developing infant, or to release the impaired part. It is desirable to complete all surgery before the child enters school. The severe complex syndactyly could rarely be solved by one operation. Normal appearance and function are rarely attained and, with growth, newer problems of imbalance and contracture appear. While rehabilitation of the hand is seldom needed for the children with cutaneous syndactyly, the severe deformities associated with complex syndactyly often require supervision in order to regain the potential function and simple dynamic splints are very helpful.


Mucus-secreting carcinoid tumor in a colonic diverticulum: report of a case.

A mucus-secreting carcinoid tumor arising in a colonic (rectosigmoid) diverticulum is presented. Carcinoid tumors in colonic diverticula have not been previously reported. The presence of mucin and argentaffin granules in the same tumor cell is illustrated. The recognition of mucus-secreting carcinoid tumors is emphasized in order to avoid overtreatment of small colonic carcinoid tumors by extensive surgical procedures.


[Bactericidal dosie-activity relationships with E. coli, K. pneumoniae and Staph. aureus (author's transl)].

In investigations into the dosis-activity relationships of bactericidal antibiotics used against E. coli, K. pneumoniae and Staph. aureus four types of activity could be demonstrated. The penicillin type shows almost no improvement in bactericidal activity despite increasing the dosage above a certain level. An increase in concentration of aminoglycoside antibiotics led to a more rapid killing of the bacteria. In cefalotin there was a linear dosis-activity relationship: rising concentrations of antibiotics led to an ever increasing bactericidal effect. The paradoxical bactericidal effect ("Eagle effect'') described for penicillin G with enterococci could not be demonstrated.


Regulation of thyrotropin (TSH) release and production in monolayer cultures of transplantable TSH-producing mouse tumors.

Studies of TSH release and production were performed in short term monolayer cultures of transplantable, thyroid hormone responsive, thyrotropin (TSH) producing mouse pituitary tumors. These tumors contained large amounts of TSH, small amounts of growth hormone (GH) and no detectable luteinizing hormone (LH), indicating that the predominant hormone product of tumor cells was TSH. The TSH content per tumor cell was similar to that of the normal pituitary where thyrotrophs represent a small fraction of the total cells, suggesting that the TSH content per tumor cell was less than that of the normal thyrotroph. There was a time dependent release and production of TSH by tumor cells in monolayer culture. Thyrotropin releasing hormone (TRH) increased the release into the media and the production of TSH in a dose dependent manner. Maximum effects were noted at 0.2 ng/ml. Thyroid hormones and somatostatin inhibited both basal and TRH induced effects on both TSH release and production. TSH release as induced by TRH was calcium dependent. TSH release was stimulated by ouabain (10(-3)M) and potassium (57 mM), agents known to promote cellular calcium uptake in a calcium dependent manner. These studies indicate that tumor derived cells function in monolayer culture in a similar fashion to normal thyrotrophs. Studies were conducted to test the hypothesis that TRH action is mediated by adenosine 3',5' monophosphate (cAMP). Dibutyryl cAMP (6 mM) and theophylline (10 mM) increased TSH release suggesting that cAMP is involved in TSH release. However, TRH had no detectable effect on tumor cell adenylate cyclase activity or levels of cAMP. In contrast, PGE1 (1-10 mug/ml) stimulated adenylate cyclase activity and elevated cellular levels of cAMP without increasing TSH release. Thus, we are unable to confirm the postulate that cAMP is the intracellular mediator of TRH action.


Hormonal regulation of renal ornithine decarboxylase activity in the rat.

The regulation of the activity of the renal enzyme ornithine decarboxylase (L-ornithine carboxy-lyase, EC 4.1.1.17) was examined in the rat. In the intact animal adapted to a light/dark cycle of 14 hours and 10 hours, respectively, the level of renal ornithine decarboxylase activity was rhythmical and paralleled the diurnal rhythm in plasma corticosteroid concentration. Renal ornithine decarboxylase activity and plasma corticosterone were highest during the early hours of darkness and lowest during the hours of light. Following hypophysectomy, the level of renal ornithine decarboxylase activity declined rapidly and remained low and without a demonstrable diurnal rhythm. When pituitary hormone levels were temporarily restored in the hypophysectomized rat by the injection of pituitary extract, renal ornithine decarboxylase activity increased rapidly, reached a peak within 8 hours, and returned toward pre-injection levels by 12 hours. Exogenous growth hormone, ACTH and cortisol each increased renal ornithine decarboxylase activity in the hypophysectomized rat, with the highest levels of activity being achieved with growth hormone. Other pituitary hormones (FSH, LH, TSH and prolactin) were ineffective. After bilateral adrenalectomy, renal ornithine decarboxylase activity retained a rhythmical pattern similar to that observed in the intact rat, but the levels were increased. Growth hormone and cortisol increased renal ornitine decarboxylase activity in the adrenalectomized-hypophysectomized animal to the same extent as in the hypophysectomized animal, but ACTH was almost totally ineffective. These data suggest that the pituitary plays a major role in the regulation of renal ornithine decarboxylase activity in the rat, primarily through the rhythmical secretion of growth hormone and ACTH.


Actions of pregnant mare serum gonadotropin in the immature female rat: correlative changes in blood steroids, gonadotropins, and cytoplasmic estradiol receptors of the anterior pituitary and hypothalamus.

Several blood steroids, serum gonadotropins and cytosol estradiol receptors of the anterior pituitary and hypothalamus were quantified in immature female rats which were induced to ovulate with pregnant mare's serum gonadotropin (PMSG). Studies revealed that serum levels of progesterone, 17-hydroxyprogesterone, testosterone, androstenedione and estradiol were initially elevated at 6 PM (day 30) after administration of 8 IU of PMSG at 10 AM day 30. Serum levels of estradiol and testosterone rose progressively from day 30 through the AM of day 32. A further increase in serum concentrations of progesterone, 17-hydroxyprogesterone, androstenedione, testosterone, and dehydroepiandrosterone occurred on the PM of day 32 whereas serum estradiol levels declined. Serum levels of all steroids declined on the day of estrus (33) and only progesterone levels were further elevated on day 34 (diestrus). Dihydrotestosterone concentrations were minimally altered by PMSG treatment. Saline administration resulted in no significant alterations in levels of any steroid quantified from day 29 to 34 in control animals. A progressive decline in cytosol estradiol receptor content of the anterior pituitary and hypothalamus was documented following PMSG treatment of intact female rats; there was no depletion of receptors following PMSG administration to ovariectomized immature rats. Maximal depletion of cytosol estradiol receptors occurred on day 32 with replenishment of cytosol estradiol receptor levels on estrus (day 33). The preovulatory gonadotropin surge was found to occur on the PM of day 32 after maximal receptor depletion. The cycle of depletion and replenishment of receptors was repeated during a second spontaneous estrous cycle four days later which coincided with a rise and fall in serum estradiol levels. It is suggested that the depletion of cytosol estradiol receptors of the anterior pituitary/hypothalamic unit may be causally related to the preovulatory gonadotropin surge resulting from PMSG administration to immature female rats. In addition, changes in blood steroids and gonadotropins after PMSG treatment are similar to those reported for proestrus-estrus-diestrus I of the normal adult estrous cycle. These findings further demonstrate the validity of the PMSG-primed immature female rat preparation as a model for the estrous cycle of the adult rat.


Binding of human chorionic gonadotropin and response of cyclic nucleotides to luteinizing hormone in luteal tissue from rats treated with prostaglandin F2alpha.

Prostaglandin F2alpha (PGF2alpha) caused a marked and highly reproducible fall of over 85% (P less than .001) in serum progesterone and in the capacity of luteal tissue to bind hCG, within 30 h. No change in the affinity of the receptor for iodinated hCG was observed. The addition of LH (1 mug/ml) to incubation flasks containing luteal tissue slices from both control and PGF2alpha-treated animals caused a significant stimulation of progesterone output in the control (P less than .05), but not in tissue previously exposed to PGF2alpha in vivo. Indeed, progesterone output by the latter tissue was severely reduced independent of the presence of LH. Although cAMP content was significantly elevated in luteal tissue incubated with LH (P less than .01), the degree of stimulation of cAMP by LH was reduced 35% (P less than .05) in luteal tissue from rats treated with PGF2alpha. The content of cGMP was generally reduced by addition of LH to the incubation media, by pretreatment of the animals with PGF2alpha, and by incubation alone. This study shows that luteolysis induced by PGF2alpha is accompanied by a loss in gonadotropin receptor, a conclusion supported by the loss in LH stimulation of cAMP and progesterone synthesis.


Immunoreactive alpha-MSH and ACTH levels in rat plasma and pituitary.

A radioimmunoassay is described for the measurement of alpha-melanocyte-stimulating hormone (alpha-MSH). The antibody was produced in rabbits by immunization with alpha-MSH coupled to bovine serum albumin with carbodiimide. The antibody did not react significantly with ACTH, beta-MSH, or 6 fragments of ACTH. The sensitivity and reliability of the assay were improved by employing a simple plasma extraction procedure. When applied to a 2 ml plasma sample, the detection limit of the radioimmunoassay was 6 pg/ml. ACTH was measured with a sensitive and specific radioimmunoassay previously described for humans and adapted for the rat. The anti-ACTH serum cross-reacted with the biologically active portion of alpha-p ACTH and not with alpha-MSH, beta-MSH or the alpha-p 17-39 and alpha-p 25-39 fragments of ACTH. The detection limit was 20 pg/ml. Plasma and pituitary alpha-MSH and ACTH had the same immunoreactivity as synthetic alpha-MSH and ACTH. alpha-MSH and ACTH contents of the rat neurointermediate lobe were 1398 +/- 360 (SE) ng and 28.2 +/- 2.9 ng, respectively, while in the anterior lobe they were 102 +/- 31 ng and 551 +/- 36 ng, respectively. The plasma alpha-MSH concentration at 8 AM in male rats was 64 +/- 8 pg/ml when the plasma ACTH concentration was 92 +/- 15 pg/ml. Over a 24-hour period two peaks of plasma alpha-MSH were observed, one at 4 AM (142 +/- 35 pg/ml) and the other at 4 PM (139 +/- 26 pg/ml). Plasma ACTH was higher at noon (151 +/- 43 pg/ml) and 4 PM (130 +/- 48 pg/ml). Short-term exposure to ether induced a transient increase in alpha-MSH level 5 min later and a rapid return to normal levels. Plasma ACTH increased significantly 2.5 min after the onset of ether stress and remained high for 30 min. Two hours' exposure to ether did not change plasma alpha-MSH, although a 3-fold increase in plasma ACTH was observed. Haloperidol injection was followed by a large increase in plasma alpha-MSH, whereas ACTH levels increased similarly after saline and Haloperidol injection. Corticoid administration reduced ACTH, but not alpha-MSH. Three weeks after adrenalectomy, alpha-MSH levels had not changed but ACTH levels had increased ten-fold. These data indicate that alpha-MSH is secreted in the rat, and that the regulation of its secretion is different from that of ACTH.


Effects of pinealectomy on adrenal function in vivo and in vitro in female rats.

Female rats were ovariectomized and either sham-pinealectomized or pinealectomized on day 24 of age and exposed to a photoperiod of 12 h light: 12 h dark. In vivo and in vitro measures of adrenocortical function were made at 6, 7, and 8 weeks of age (18, 25, and 32 days post-pinealectomy, respectively). Pinealectomy diminished the post-castration rise in adrenal 5alpha-reductase activity in all age groups (P less than .05). The proportionate secretion of corticosterone (compared with the total steroid output) by adrenal slices was likewise enhanced (P less than .05) although the secretion and production of corticosterone in vitro was not altered. Pinealectomy substantially diminished (P less than .05) the in vivo secretion rates of reduced metabolites of corticosterone (dihydrocorticosterone and tetrahydrocorticosterone) without altering corticosterone secretion. Consequently, proportionate secretion of corticosterone in vivo was also enhanced (P less than .05). These findings are consistent with the notion that in ovariectomized rats removal of the pineal gland diminishes adrenal 5alpha-reductase activity without affecting ACTH secretion. However, in rats with ovaries intact, estrogen modified the effects of pinealectomy. Not only was the intra-adrenal metabolism of corticosterone diminished (higher proportionate output), but also resting levels of plasma corticosterone (P less than .01), corticosterone production in vitro (P less than .05), and total adrenal steroidogenesis in vitro (P less than .01) were increased. Thus, ACTH secretion may be enhanced following pineal removal in the presence of estrogen. The data suggest that the pineal gland, together with the ovaries, plays a role in the modulation of adrenal steroidogenesis.


Temporal relationships between hormone receptor binding and biological responses in the uterus: studies with short- and long-acting derivatives of estriol.

The temporal relationships between hormone receptor binding and early and late biological responses in the uterus were examined using estriol (E3), a weak estrogen, and several more long-acting estriol derivatives, namely ethinyl estriol (EE3), estriol cyclopentyl ether (E3CPE), and ethinyl estriol cyclopentyl ether (EE3CPE). Dose-response curves of 3-day uterotrophic assays indicate that biological potency follows the order EE3CPE greater than EE3 or estradiol greater than E3CPE greater than E3. After a single injection of 5 mug of compound, E3 elicits the early uterotrophic responses (increased uterine wet weight and 2-deoxyglucose phosphorylation at 2-6 h) but gives only weak stimulation of later uterotrophic responses (enhanced rates of 2-deoxyglucose phosphorylation at 20-24 h and increased DNA synthesis rate and uterine weight over a 72 h period). E3, EE3, and estradiol all elicit a rapid (maximal by 1/2-1 h) uptake of receptor into the nucleus and show an equivalent wet weight response at 3 h. After E3, nuclear receptor levels and uterine weight decline rapidly; however, after EE3 or estradiol, nuclear receptor levels decline less rapidly remaining at least two-fold above the control until 24-48 hr, and uterine weight also remains elevated for at least 48-72 h. EE3CPE elicits both the early (4 h) and later (20-24 h) waves of glucose metabolism, shows a prolonged effect on DNA synthesis rate, and shows the most dramatic and prolonged (beyond 72 h) maintenance of elevated uterine weight and high nuclear receptor (beyond 24 h). Thus, chemical modifications of the estriol molecule which result in a prolonged stimulation of uterine growth and metabolism also result in a long-term maintenance of hormone-receptor complex in the uterine nucleus. These studies give strong support to the concept that true uterine growth requires the direct and prolonged influence of the nuclear estrogen-receptor complex.


Characterization of an inhibitor for luteinizing hormone receptor site binding.

An inhibitor for luteinizing hormone (LH) receptor site binding has been partially purified from aqueous extracts of ovaries from pseudo-pregnant or pregnant rats. The LH receptor binding inhibitor (LH-RBI) was heat-resistant, partially inactivated by trypsin or pronase digestion, and had a molecular weight of approximately 3800 daltons. The LH-RBI was not found in the ovary of mature non-pregnant rats or immature rats, nor was it found in testis or liver extracts. The LH-RBI strongly inhibited the in vitro binding of 125I-labeled ovine LH to ovarian LH receptors but did not inhibit 125I-labeled ovine prolactin binding to ovarian PRL receptors.


Hormonal responsiveness of adenylate cyclase activity in cartilage.

Adenylate cyclase activity was measured in a crude particulate fraction of hyaline cartilage obtained from the xiphoid process of the rat. Bovine parathyroid hormone (PTH) at concentrations as low as 1.3 x 10(-7)M and porcine calcitonin (CT) at concentrations as low as 2.3 x 10(-5)M significantly increased adenylate cyclase activity. Glucagon, prostaglandin E1 (PGE1) and E2 (PG2), and epinephrine at concentrations of 10(-5)M also increased activity, whereas, no increased activity was seen with the additions of somatotrophin (10 mug/ml), PGF1alpha, PGF2alpha, or T3 at 10(-5)M. The combination of doses of PTH and CT, which individually produced maximal responses, was not additive. These data provide evidence that cartilage in growing rats responds directly to PTH and CT.


Steroid formation and differentiation of cortical cells in tissue culture of human fetal adrenals in the presence and absence of ACTH.

Steroid secretion and ultrastructural differentiation of human fetal adrenal cortical cells were analyzed in tissue culture with and without ACTH. The unconjugated and sulfated endogenous neutral steroids were analyzed by gas-liquid chromatography and gas chromatography-mass spectrometry. A fetal pattern of neutral steroids, including high concentrations of sulfate conjugates, was found during the first five days of the cultivation. At 6 to 11 days of cultivation, a decrease was seen in concentrations of these steroids. However, when stimulated with ACTH, an increasing amount of steroids was secreted during days 6 to 11 and their pattern was transformed into the adult type with a 30-200 times higher secretion rate of cortisol. Cortical cells capable of proliferation in the culture had the ultrastructure of the permanent zone cells of the fetal adrenal or adult zona glomerulosa type. ACTH stimulation induced a differentiation of these cells into zona fasciculata type. The results suggest that ACTH is the main hormonal regulator in the genesis of the adult human adrenal cortex and that there is a factor during fetal life which inhibits the synthesis of the 3beta-hydroxysteroid dehydrogenase system.


A specific FSH receptor in rat granulosa cells: properties of binding in vitro.

The specific binding of [125I]iodoFSH to granulosa cells collected from immature, hypophysectomized, DES-treated rats, was studied in vitro. Specific binding occurred after 5 min and reached maximum after 3 h of incubation at 37 C. Non specific binding was very low (less than 10% of the total binding). The [25I]iodoFSH remained tightly associated with the receptor at pH 7.5, but was rapidly dissociated at pH 5. Unlabeled hFSH competitively inhibited [125I]iodoFSH binding. Kinetic analyses of equilibrium binding experiments gave an apparent association constant (Ka) of 1.34 (+/- 0.31) x 10(10)M-1 [mean (+/- SE)] and a number of binding sites per cell of (NB) 1, 130 +/- 70 (mean +/- SE). Rat prolactin, wheat germ agglutinin, and concanavalin-A did not compete with [125I]iodoFSH, but hLH, hCG, and rTSH competed at doses 300- to 900-fold higher than those of hFSH. Granulosa cells isolated from adult DES-treated rats, as well as cells collected from medium and preovulatory follicles of proestrous rats, gave Ka and NB values similar to those described above. A comparative study of rabbit granulosa cells indicated a much lower binding affinity compared with those from the rat.


Effects of hyperosmolarity on the cyclic AMP concentration and lipolysis of the adipocyte stimulated by adrenocorticotropic hormone.

The effects of ACTH on 3',5'-cyclic AMP (cAMP) levels and lipolysis were examined on isolated adipocytes incubated in either isosmolar or hyperosmolar media. The ability of ACTH to induce intracellular cAMP accumulation was greatly enhanced by incubating cells in hyperosmolar sucrose (100 to 400 mM) solutions. Hyperosmolar solutions prepared by the addition of either NaCL, glucose or mannitol enhanced the ACTH effect on cAMP to the same extent as did the hyperosmolar sucrose solution, but hyperosmolar urea solutions did not have such an effect. The effect of hyperosmolarity was shown only in cells stimulated by lipolytic hormones, and the effects were still evident in the presence of high concentrations of theophylline, indicating the effect of hyperosmolarity is to facilitate hormone action on the receptor-coupler system of the adipocyte membrane. The action of glucagon on cAMP was augmented much less than the actions of ACTH and isoproterenol. Basal as well as ACTH or exogenous cAMP stimulated lipolysis was lower in hyperosmolar sucrose solutions. Some mechanism by which hyperosmolarity interferes with the metabolic sequence beyond the accumulation of cAMP was suggested.


Enzymatic dissociation and short-term culture of isolated rat anterior pituitary cells for studies on the control of hormone secretion.

A convenient procedure has been developed for preparing a suspension of isolated rat anterior pituitary cells which retains responsiveness to secretagogues. Rat anterior pituitaries were dispersed with collagenase and hyaluronidase followed by mechanical dispersion by means of a Pasteur pipette. Immediately after dispersion, the cells showed only slight responses to secretagogues, whereas after short-term culture (20-22 h) in the presence of sera, the cells recovered their ability to respond to synthetic LH-releasing hormone (LHRH) and synthetic thyrotropin-releasing hormone (TRH). During a 3-h incubation, cells prepared from pituitaries of male rats released LH and FSH, or TSH and prolactin (PRL) in amounts directly related to the dose of synthetic LHRH or TRH, respectively. The minimum effective concentrations of hypophysiotropic hormones lay between 10(-10) and 10(-9)M, although it was observed that cells originating from female rats usually gave quicker and larger responses to LHRH. No significant net increase in the total hormonal content (cells + medium) of radioimmunoassayable LH or FSH in response to LHRH, or of TSH or PRL in response to TRH, was observed during the 3-h incubation period. The cells released significant amounts of PRL, TSH, and to a lesser extent, LH, in response to 1-5 X 10-3M N6,O2'-dibutyryl cyclic AMP, accompanied by remarkable elevation in total content (cells + medium) of PRL and TSH but not of LH. The response of the cells to theophylline or high [K+] was similar to that usually observed in previous hemipituitary experiments. These results demonstrate the viability of this in vitro cell system and its suitability for further study of the regulation of the secretion of pituitary hormones.


Biochemical actions of follice-stimulating hormone in the sertoli cell of the rat testis.

The sequenc of biochemical events associated with the action of follicle-stimulating hormone (FSH) in the testis has been investigated using a Sertoli cell-enriched testis model system. The Sertoli cell-encriched testis, created by irradiation of male rats in utero, is devoid of germinal elements but contains a normal complement of supportive Sertoli cells. Comparison of the Sertoli cell-enriched testis with normal testis, demonstrates that the two types of testes contain equal numbers of FSH specific receptors, judged by the binding of labeled hormone. In addition, FSH over a concentration range from 6 X 10(-11) to 6 X 10(-9)M will stimulate the production of adenosine 3',5' monophosphate (cAMP) in the Sertoli cell-enriched testis in a manner indistinguishable from that of the normal testis. Incubation of Sertoli cell enriched testis also results in the activation of soluble cAMP-dependent protein kinase. This response to FSH is dependent upon the age of the animal and disappears at about 32 days of age. While sensitivity to the hormone can still be detected in mature Sertoli cell-enriched animals by the addition of the phosphodiesterase inhibitor 1-methyl-3-isobutyl-xanthine, no detectable increase in phosphodiesterase activity is apparent after 30 days of age. Injection of FSH into Sertoli cell-enriched animals results in an increase in total testicular protein  synthesis as well as in the production of the Sertoli cell-specific protein, androgen-binding protein within 30 minutes. Furthermore, while hypophysectomy of Sertoli cell-enriched animals result in a decline of the testicular concentration of androgen-binding protein, the injection of FSH will stimulate and maintain the levels of androgen-binding protein in such animals. These results demonstrate that the Sertoli cell-enriched testis is capable of carrying out the sequence of biochemical events previously described for FSH in the normal testis and therefore, suggest that the Sertoli cell is the primary target cell for FSH action.


Control of uterine estrogen receptor levels by progesterone.

The mechanism by which progesterone antagonizes estrogenic stimulation of uterine growth was examined in the immature rat. Rats received daily injections of 2.5 mug estradiol (E) for 2 days and on day 3 either 2.5 mug E or 2.5 mug E plus 2.5 mg of progesterone (P). The quantity of nuclear and cytoplasmic estrogen receptor was determined by [3H]estradiol exchange at various intervals after injection of E or E + P. In both groups, nuclear receptor estrogen complex (RnE) increased dramtically one hour after injection and showed a gradual decline from 4 to 24 h after injection. The quantity of cytoplasmic receptor, Rc, decreased to low levels by one hour and began a gradual increase from 4 to 8 h in both groups. However, between 8 and 24 h after injection, the level of Rc continued to increase in the E treatment group (2.39 +/- 0.21 pmol/uterus at 24 h) but remained at the 8 h level in the E + P group (1.09 +/- 0.04 pmol/uterus at 24 h). This observation suggests that two seperate processes are involved in the replenishment of Rc and that progesterone inhibits the second phase of replenishment. The binding affinity and specificity of Rc for estrogens following E + P pretreatment were identical to those of the E pretreatment group. Therefore, P does not alter the binding properties but rather the intrauterine level of Rc. Treatment with E on day 4, when Rc levels differ between E and E + P groups, stimulated uterine weight and protein content on day 5 in the E pretreatment group. However, minimal stimulation was observed in the E + P pretreatment group. The quantity of RnE and the time of nuclear retention of RnE following E injection on day 4 was greater in the E group than in the E + P group. The effect of progesterone on Rc replenishment was dose-dependent (range, 0.1-2.5 mg; 1/2 maximal, 0.5 mg). Injection of testosterone propionate (1.0 mg), a weak estrogen antagonist, with E on day 3 resulted in slightly reduced levels of Rc on day 4. This reduction also correlated with a reduced sensitivity to treatment with E on day 4. These data, together with previous studies from our laboratory, suggest that progesterone and other estrogen antagonists such as nafoxidine and testosterone propionate inhibit estrogen action by interfering with the replenishment of Rc, thereby reducing the number of receptor estrogen complexes that are translocated and retained by uterine nuclei.


Effects of ACTH, dexamethasone, adrenalectomy and stress on cAMP content of adenohypophysis and adrenals in rats.

The effects of adrenalectomy, ether-laparotomy stress and in vivo administration of either ACTH or dexamethasone on the cAMP levels in the anterior pituitary and the adrenal glands were investigated in male rats. After adrenalectomy or ether-laparotomy stress, an increase of the pituitary cAMP levels was observed. A prior administration of dexamethasone failed to inhibit the increase of the pituitary cAMP levels. Acute administration of dexamethasone increased the cAMP levels in the pituitary. Chronic administration of dexamethasone decreased the cAMP levels in the pituitary, but ACTH did not. These data suggest that cAMP might be involved in the mediation of the hypothalamo-pituitary-adrenal axis.


A receptor site for prolactin in lactating mouse mammary tissues.

Prolactin iodinated by lactoperoxidase method showed immunologically, electrophoretically and biolo9gically similar properties to native prolactin and possessed enough specific radioactivity for receptor studies. 1251-prolactin was incubated with mouse mammary tissues at 8 days of lactation. Both binding and release of 1251-prolactin depended on incubation time and temperature and were maximal at 37 degrees C. Michaelis constant was estimated to be 1.4 X 10(-9) M from Lineweaver-Burk plot and to be 1.2 X 10(-9) M from id-value of the dose-response curve for displacement with native prolactin. Total number of binding sites for prolactin was 1.38 X 10(-15) mole per mg weight of tissue. Ovine prolactin, human growth hormone and human placental lactogen complete with 1251-prolactin and dose-response curves for these three hormones were all parallel. These results suggest the existence of a specific receptor site with high affinity for prolactin in lactating mouse mammary glands.


Effects of calcium ionophore (A-23187) on glucose oxidation and iodide transport in dog thyroid slices.

A calcium ionophore (A-23187, 20 mug/ml) stimulted 14C-1-glucose oxidation in dog thyroid slices to an extent equivalent to that obtained by the optimal concentration of dibutyryl cyclic AMP (1mM). Furthermore, the ionophore augmented the stimulation by dibutyryl cyclic AMP much more than the simple additive effect. The ionophore also enhanced the effect of TSH, but to a lesser extent. Under conditions where organic binding was blocked, T/M ratio of radioiodine concentration was lowered in slices by the ionophore; the findings similar to those obtained with TSH and dibutyryl cyclic AMP. The ionophore exhibited a slightly depressive effect on the basal cyclic AMP level. The elevation by TSH of cyclic AMP levels was also slightly depressed by the ionophore, but statistically insignificant in most cases. These results indicate that calcium ion may play an important role in the TSH regulation of iodide transport and glucose metabolism in the thyroid, in some cases by augmenting the effects of cyclic AMP.


5alpha-Dihydrotestosterone binding protein in rat ventral prostate; purification, nuclear incorporation, and subnuclear localization.

Treatment of cytosol from the rat ventral prostate with cold acetone (-20 degrees C) evoked a 8 approximately 10-fold increase in the binding capacity with 5alpha-dihydrotestosterone (DHT). Starting from the extract of acetone-dried prostate cytosol, some 400 approximately 600-fold purification of the DHT-binding protein complex was acieved by (NH4)2504 fractionation, DEAE-cellulose chromatography and gel- filtration with Sephadex G-200. The purified 3H-DHT-binding protein complex was incorporated into the nuclei from the ventral prostate in a temperature dependent manner. The similar incorporation was also observed in nuclei from the liver and the kidney...


Further experimental results concerning the relationship of muscular exercise and adrenal function.

The relationship between muscular work and adrenal function was investigated by using daily swimming exercise in the rat. 1. Adrenal in vitro sensitivity to ACTH was found to have increased after six weeks of swimming training 2. In the development of adaptation the main part was played by muscular exercise itself, since a conditioning of the animals merely to the circumstances of swimming failed to stimulate such adaptive changes as seen in the course of regular physical training. 3. Plasma steroid response to certain stressor agents, such as histamine or acute exhaustion (decrease), respectively to ACTH administration (increase) was similar in ACTH-pretreated and regularly exercised animals. 4. In the 6th to 12th weeks of exercise the trained animals developed a lower plasma steroid content at rest than the controls. This feature was retained even four weeks after the cessation of training sessions. Experimental physical training reduced post-exercise steroidemia in the period of adaptation, that is, after the 6th week. After a pause of 4 weeks, however, the post-exercise steroidemia approached again the control values.


Influence of thyrotropin releasing hormone and dibutyryl cyclic adenosine 3',5'-monophosphate on ultrastructure of adenohypophysis in organ culture.

Ultrastructure of adenohypophysis in organ culture exposed to thyrotropin releasing hormone or dibutyryl cyclic adenosine 3',5'-monophosphate has been investigated. The features of increased secretory activity of glandular cells as well as the occurrence of a strongly osmiophilic substance in the intercellular spaces was observed in cultures exposed to both investigated factors.


Evocability of a slight positive oestrogen feedback action on LH secretion in castrated and oestrogen-primed men.

Orchidectomized heterosexual men suffering from prostatic cancer with suppressed serum LH values by oestrogen priming exhibited a slight, but significant surge of LH secretion following oestrogen injection. In contrast, orchidectomized and oestrogen-primed heterosexual men with unsuppressed serum LH levels as well as intact heterosexual men did not display any positive oestrogen feedback action on LH secretion,  as it was observed in intact homosexual men.


Virus association with wastewater solids.

The solids produced at an urban wastewater treatment plant, tested semiquantitatively, almost all contained human intestinal viruses. Reoviruses and five or more types of enteroviruses were present. Sludge, digested anaerobically at 30 degrees-32 degrees C, and grit contained measurable levels of viruses. Until reliable means of inactivating the viruses have been developed and implemented, great care should be taken in disposing of these solids.


Accumulation of cadmium in rat liver cadmium binding protein following single and repeated cadmium administration.

The accumulation of cadmium in rat liver cadmium binding protein induced by single and repeated intraperitoneal injections of CdCl2 and the de novo biosynthesis of CdBP were studied by using 109Cd to measure cadmium binding in the CdBP and 35S incorporation as indicator of protein synthesis. The biosynthesis of CdBP is controlled by the cadmium concentrations. For single doses up to 1 mg Cd2+/Kg b.w. about 50% of the cadmium is present in the soluble fraction of liver bound to CdBP and the incorporation of 35S-cysteine is linear with the cadmium concentration. When single doses ranging from 1 to 3 mg Cd2+/Kg b.w. are administered the fractions of both 35S-cysteine and cadmium incorporated into de novo synthesized CdBP gradually decrease. For single doses higher than 3 mg Cd2+/Kg b.w. the biosynthesis capability is maximum and 20 mug Cd/g liver can be incorporated into the de novo biosynthesized CdBP. When rats are treated every day with amounts of cadmium of about 0.8 mg Cd2+/Kg b.w. for up to 8 days a dose-proportional increase in both Cd incorporation and CdBP biosynthesis are observed. This shows a cumulative incorporation of cadmium in the de novo biosynthesized CdBP. Experiments carried out by injecting 65ZnCl2 and 203HgCl2 every day showed that they are not accumulated like cadmium and do not induce the biosynthesis of rat liver CdBP after repeated administration over 7 days.


The reduction of plastocyanin by plastoquinol-1 in the presence of chloroplasts. A dark electron transfer reaction involving components between the two photosystems.

The reduction of plastocyanin by plastoquinol-1 was efficiently catalysed by disrupted chloroplasts or etioplasts in the dark. The reaction was inhibited by 2,5-dibromomethylisopropyl-p-benzo-quinone which inhibits photosynthetic electron transport between plastoquinone and cytochrome f. Evidence is presented that the reduction took place via cytochrome f, and that plastoquinone-9 was not involved. Triton X-100 and organic solvents were inhibitory, but partial fractionation was achieved without loss of activity by density gradient centrifugation in the presence of high digitonin concentrations. All active material contained cytochromes b-559LP and b-563 in addition to cytochrome f, but these b-type cytochromes were not directly involved. Other 1-electron acceptors could be used in place of plastocyanin, for instance ferricyanide and Pseudomonas cytochrome c-551. The reaction can be applied to give a sensitive dark assay for active cytochrome f. It is suggested that cytochrome f possesses two sites for interaction with redox reagents: a hydrophilic site with which plastocyanin reacts by electron transfer and a hydrophobic site with which plastoquinol reacts by hydrogen atom transfer.


Brown adipose tissue mitochondria: recoupling caused by substrate level phosphorylation and extramitochondrial adenosine phosphates.

1. Uncoupled oxidative phosphorylation in isolated guinea pig brown-adipose-tissue mitochondria is reflected by a low phosphorylation state of adenosine phosphates in the mitochondrial matrix and in the extramitochondrial space during oxidation of succinate or glycerol 1-phosphate in the presence of serum albumin and 100 muM ADP. Recoupling of respiration and phosphorylation in the mitochondria is indicatdd by a dramatic increase in the phosphorylation state of adenine nucleotides in both compartments, when substrates inducing substrate level phosphorylation are respired. In this case ATP/ADP ratios in the extramitochondrial compartment are 10-15 times higher than in the mitochondrial matrix. 2. Recoupling mediated by substrate level phosphorylation depends on the presence of extramitochondrial adenosine phosphate and on intact adenine nucleotide translocation. In the presence of substrate level phosphorylation the amount of extramitochondrial ADP required to restore energy coupling can be extremely low (20 muM ADP or 10 nmol ADP/mg mitochondrial protein respectively). If substrate level phosphorylation is prevented by rotenone or in the presence of atractyloside, 20-50 times higher amounts of extramitochondrial adenine nucleotides are necessary to cause coupled oxidative phosphorylation. The recoupling effect of ATP is significantly stronger than that of ADP. 3. GDP (100 muM) causes a rapid increase of the ATP/ADP ratio in both compartments which is independent of substrate level phosphorylation as well as of the extramitochondrial adenosine phosphate concentration and the adenine nucleotide carrier. 4. The amount of extramitochondrial adenosine phosphate in guinea pig brown-adipose-tissue (18 nmol/mg mitochondrial protein or 2.5 mM respectively) would suffice for recoupling of oxidative phosphorylation mediated by substrate level phosphorylation under conditions in vitro; this suggests that substrate level phosphorylation is of essential importance in brown fat in vivo with respect to energy conditions in the tissue during different states of thermogenesis.


New coenzymically-active soluble and insoluble macromolecular NAD+ derivatives.

Reaction in dimethyl sulfoxide of nicotinamide 8-bromoadenine dinucleotide with the disodium salt of 3-mercaptopropionic acid afforded nicotinamide-8-(2-carboxyethylthio)adenine dinucleotide, a new NAD+ analogue functionalized at the adenine C-8 position by an omega-carboxylic side chain. Carbodimide coupling of the latter derivative to high-molecular-weight water-soluble (polyethyleneimine, polylysine) and insoluble (aminohexy)-Sepharose) polymers gave the corresponding macromolecular NAD+ analogues. These derivatives have been shown to be enzymically reducible. The polyethyleneimine analogue showed a substantial degree of efficiency relative to free NAD+ with yeast alcohol dehydrogenase (47%) but a considerably lower one with rabbit muscle lactate dehydrogenase (3%); the polylysine analogue showed a low degree of efficiency with both enzymes (5-6%).


Kinetics of adenosine 3':5'-monophosphate accumulation in dog thyroid slices.

Dog thyroid slices have been stimulated in vitro by thyrotropin. The kinetics of adenosine 3':5'-monophosphate (cyclic AMP) accumulation due to the activation of adenylate cyclase were measured. Experimental results have been interpreted by means of a numerical simulation based on a theoretical description of the overall process which consists of three steps: (a) the penetration of thyrotropin in the slices, (b) the binding of the hormone to the specific receptor and the consequent activation of adenylate cyclase, (c) the kinetics of cyclic AMP accumulation in each cell due to the interplay between synthesis and degradation. The numerical values of the parameters needed for the simulation were deduced from separate experiments. Diffusion of thyrotropin in the slices has been calculated from the kinetics of efflux of [3H]sucrose, and with or without albumin. Adenylate cyclase activity and activation by thyrotropin in homogenates and purified membranes were measured. Binding experiments of cyclic AMP on crude extract of dog thyroid lead to the conclusion that the maximal capacity of the specific binding site is close to the cyclic AMP content in resting thyroid cells. The purpose of this paper is to verify the validity of the current concepts about the mechanisms taking place in this process by comparing experimental results and theoretical simulation. It results from the study that for this system the time between the activation of adenylate cyclase and half maximal cyclic AMPaccumulation is equal to 3 min 45 s and the pool of cyclic AMP is renewed three times per minute. It has been also stressed that the thickness of the slices as well as inhibitors of phosphodiesterases change the apparent kinetics of cyclic AMP accumulation.


Cerebrospinal fluid concentration of cyclic AMP in cerebrovascular diseases.

Concentration of cyclic adenosine 3',5'-monophosphate (cAMP) and activities of some enzymes were measured in cerebrospinal fluid (CSF) from 38 patients with various cerebrovascular diseases. Cerebral infarction of the carotid area (less than 14 days after the attack) revealed a significant increase in CSF cAMP level in comparison to a transient ischemic attack (TIA) and cephalagia without any pathological findings (control group). A trend towards elevated values was observed also in cerebral hemorrhage, whereas the CSF cAMP concentrations in subarachnoid hemorrhage, TIA, syncope, and cerebral infarction of at least 2 months of duration were in the range of control values. A significant rise in CSF enzyme activities was observed only in hemorrhagic disorders. Hypertensive patients with TIA showed significantly higher CSF cAMP values than normotensive ones. A similar positive correlation between blood pressure and CSF aAMP concentrations was found also in subarachnoid hemorrhage and syncope groups. On the basis of the present results it is suggested that in cerebrovascular diseases CSF cAMP concentration reflects the size and the time of the destruction of cerebral cells, and correlates with hypertension of the patient possibly indicating an increased sympathetic activity.


Olfaction in multiple sclerosis. With a note on the discrepancy between optic and olfactory involvement.

Serial binary dilutions of amyl acetate and nitrobenzene were used in a double-blind test to determine olfactory acuity of 40 multiple sclerosis (MS) patients. Similar determinations were made on 24 age- and sex-matched controls. Results of repeated measurements on 18 patients indicated that the methodology was reliably reproducible. When mean olfactory threshold on MS patients was compared with that of the controls no differences could be found. Among the MS patients, olfactory threshold did not vary with the presence or absence of optic involvement and correlated poorly with the degree of disability. These results are in disagreement with those of WENDER and SZMEJA who reported olfactory dysfunction in 18 out of 52 MS patients. Our findings are compatible with the observations of ZIMMERMAN and NETSKY who failed to find any plaques in olfactory tracts of MS brains. The significance of this disparity between involvement of optic and olfactory systems, both of which contain central myelin, is discussed. Available biochemical evidence suggests that, as postulated by LUMSDEN, sparing of olfactory tracts may in part be related to the relatively lower basic protein content of these structures.


Effect of convulsions and anticonvulsive drugs on cerebrospinal fluid cyclic AMP in rabbits.

Cyclic adenosine 3',5'-monophosphate (cAMP) concentration and aspartate amino transferase (ASAT or GOT) activity were measured from cerebrospinal fluid (CSF) of rabbits before and after electrically induced convulsions. The convulsions caused a significant increase in CSF cAMP concentration. The treatment with phenobarbital, diphenylhydantoin and carbamazepine for 7 days decreased the basal cAMP values in CSF and partly inhibited the rise after the convulsions. The alterations in ASAT activity were parallel, but less striking. The present results suggest the involvement of cAMP in epileptic discharge and in the mechanism of action of anticonvulsants.


Studies on the biochemical aspects of the 'disulfiram-like' reaction induced by oral hypoglycemics.

In vitro experiments, using rat liver homogenates, were designed to examine certain of the proposed enzymatic mechanisms for the interaction of oral hypoglycemic drugs with monoamine and ethanol metabolism. The oxidative degradation of tryptamine was studied by measuring indoleacetic acid (IAA) production and conclusions were drawn with regard to the activity of monoamine oxidase, aldehyde dehydrogenase and ethanol dehydrogenase. Acetohexamide, hydroxyhexamide, tolazamide, tolbutamide and chlorpropamide failed to reveal any specific inhibition of the three enzymes. Ethanol (0.2% w/v) and disulfiram decreased IAA formation, as did a lack of available aldehyde dehydrogenase and NAD, but these reductions were not enhanced by the hypoglycemic agents. The results suggest that the 'disulfiram-like' reaction which occurs in certain patients imbibing ethanol while receiving oral hypoglycemic drugs, depends upon some factor(s) other than, or additional to, a specific interference with monoamine and/or ethanol metabolism.


The effect of imidazole on the isometric contractility of the isolated hemidiaphragm of the rat.

Imidazole (0.73-15.9mM) was found to increase both tension developed Td and the maximum rats of rise of tension, dT/dtmax, in the isolated hemidiaphragm of the rat during indirect stimulation. Similar effects were obtained during direct stimulation and in the presence of d-tubocurarine. Imidazole (0.73-22 mM) antagonized the action of d-tubocurarine. This effect was particularly pronounced in preparations pretreated with imidazole. Propranolol did not significantly change the action of imidazole on Td and dT/dtmax during direct stimulation. Similarly, propranolol did not affect the action of low concentrations of imidazole during indirect stimulation. When present in the bath for periods of time longer than 15 min, propranolol significantly depressed the effect of even high concentrations of imidazole on Td and dT/dtmax during indirect stimulation. Histamine (0.18-0.91 mM) did not affect either Td or dT/dtmax. In the experiments in vivo, imidazole (12.5-100 mg/kg) produced a small increase both in Td and dT/dtmax of the gastrocnemius muscle during sciatic nerve stimulation. The available evidence indicates that the action of imidazole on Td and dT/dtmax is not connected with its action on phosphodiesterase, but it is most probably due to a direct action on the muscle.


[Pathological studies on corynebacterial ulcerative entero-colitis in rats treated with ACTH (author's transl)].

Ulcerative entero-colitis was developed in 6-week-old male Sprague-Dawley rats treated daily with ACTH (4 mg/kg. s. c.) as well as necrotic purulent lesions in liver, kidney, lung or heart. Incidence of ulcerative lesions was 6.3% in Farm-A rats and 56% in Farm-B rats. Although ulcerative lesions were mostly observed in cecum, the similar lesions were also detected in distal ileum or proximal colon in some cases. Histologically, the lesions were characterized by focal necrosis demarcated from surrounding normal tissue containing a number of clumps of bacteria and cellular debris. Bacteriological examination revealed that provocation of Coryne-bacterium kutscheri by ACTH-treatment resulted in appearance of the lesions. By means of intravenous or intraperitoneal inoculation with the strain isolated from lesion similar lesions were produced in the cecum of inoculated rats under the ACTH-treatment.


Extracellular Ca2+ and Mg2+ and cellular metabolism of phospholipids.

High extracellular concentration of Ca2+ inhibits the incorporation of 32P into the cellular phospholipids. This effect is more significant in neoplastic than in normal cells, and it is accompanied by an increase of the percentual incorporation into the lecithin fraction.


[Stability of some bradykinin analogues against kininase II (author's transl)].

Some analogues of bradykinin, especially with replacements by other amino acids of phenylalanine in position 8, have been investigated for enzymatic stability against kininase II from rat duodenum microsomes and rat uterus plasma membranes, respectively. As compared with bradykinin, two of the analogues, [8-erythro-beta-phenylserine]- and [8-erythro-alpha-Amino-beta-phenylbutyric acid]-Bradykinin were stable to enzymatic degradation. Therefore, the latter may be used for studies in hormone-receptor interaction.


Penetration of phospholipases A2 and C into the squid (Loligo pealii) giant axon.

Following 1 h exposure, the level of phospholipase A2 penetration into the axoplasm of the squid giant axon was 107 to 350% of that in the external media; corresponding values for phospholipase C were 18 to 31%. Phospholipases can therefore be used to study phospholipid function in axons since they can penetrate through connective tissue and Schwann cell to reach the axolemma.


Opposite effects of vasotocin injected intrapituitarly and intraventricularly on corticotropin release in mice.

A single injection of 10 pg synthetic arginine vasotocin into the pituitary significantly potentiated adrenal hypertrophy produced in male mice by unilateral adrenalectomy, whereas the same concentration injected into the 3rd ventricle produced adrenal atrophy.


[Study of the mitotic activity during the preneoplastic period in the liver of rats treated by diethylnitrosamine (author's transl)].

The chronic administration of diethylnitrosamine in drinking water induces liver tumors in the rat. The preneoplastic period may be divided in two phases differing mainly in the loss of circadian control of mitotic activity in hepatocytes.


Free and sulfoconjugated dehydroepiandrosterone, cyclic adenosime-3',5'-monophosphate, and free estriol in maternal and cord blood.

When free DHEA, its sulfatide, and sulfate were assayed in maternal plasma as well as in umbilical cord arterial and venous plasma, rather high concentrations were found in either fraction from cord arterial plasma, reflecting the fetal contribution not only of free DHEA and DHEA sulfate, but also of the lipophile steroid sulfatide. Since high DHEA levels were associated with elevated c-AMP concentrations, a certain interrelationship of both parameters is indicated. In the course of delivery, a rapid decrease of free estriol in maternal plasma was observed. Higher concentration of free estriol in umbilical venous plasma pointed at its placental biosynthesis from fetal precursors.


Adenovirus type 12 infection of defined mouse-human hybrid cell clones.

Human adenovirus type 12 does not multiply in mouse cells; only viral T-antigen is detected. Mouse-human cell hybrid clones containing human chromosomes A3, B5, C7, C11, C12, D14, E17, F19 and F20, support synthesis of adenovirus DNA and capsid antigens.


Effects of dibutyryl cyclic AMP on cultured brain cells from chick embryos of different ages.

The effect of dibutyryl cyclic AMP on cultivated astroblasts from 8-, 12- and 15-day-old chick embryo brain has been studied. The results show that these cells must have reached some degree of maturation in ovo to become morphologically differentiated by the cyclic nucleotide.


Autoradiographic study of the penetration of non-histone chromatin proteins into differentiating cells.

Attention has previously been drawn to a specific effect of NHCP on embryonic Pleurodeles cell differentiation. With a modified NHCP labelling technique, autoradiography has revealed a cytoplasmic concentration of labelled NHCP and has not revealed any difference between homospecific and heterospecific NHCP penetration.


The influence of MuLV and SV40 viruses on senescence in mouse fibroblasts in vitro.

Mouse cells productively infected with Moloney leukaemia virus (MuLV) underwent senescence in a manner similar to control cells, although they recovered more readily as an established line. Rapidly growing cell lines were also obtained following simian virus 40 (SV40) infection of senescent cells. However, superinfection of senescent MuLV-producing cells by SV40 led to slower growing cells with a reduced output of infectious MuLV.


Matrix vesicle heterogeneity: possible morphogenetic functions for matrix vesicles.

Extracellular membranous matrix vesicles were localized and described using electronmicroscopy during chondrogenesis, osteogenesis, and dentinogenesis. Evidence indicates that matrix vesicles in each of these specific tissue types function to concentrate and transport ions and enzymes which serve as nucleation sites for the mineralization of hydroxylapatite. We have examined different developmental stages of Meckel's cartilage, alveolar bone and epithelial-mesenchymal interactions associated with tooth formation in newborn mice. These ultrastructural studies indicate matrix vesicle heterogeneity. Whereas most matrix vesicles contain alkaline phosphatase activity during cartilage, bone and dentine mineralization, in earlier developmental stages matrix vesicles contain acid phosphatase activities and little, if any, alkaline phosphatase. Tissue type, specific developmental stage, and ultrastructural criteria indicate various "classes" of matrix vesicles. During epithelial-mesenchymal interactions in tooth development, mesenchymal cells (preodontoblasts) appear to be the source of matrix vesicles as indicated by the complementarity between H-2 histocompatibility alloantigen specificity on the cell surface and that of the matrix vesicle outer surface; matrix vesicles are limited by a trilaminar membrane derived from the mesenchymal cells. Some of the vesicles located adjacent to dividing inner enamel epithelial cells contain RNA's as determined by electron microscopic autoradiography in situ, as well as by direct biochemical assays. We postulate that matrix vesicles have many different and important biological functions, one of which may be to mediate developmental information from mesenchyme to epithelia during "instructive" stages of tooth development.


[Impulse activity of neurons in the rabbit sensory-motor cortex during recruitment].

In unanesthetized rabbits, recruiting response to low-frequency stimulation of the thalamic center medial was followed by a change in the overall level of neuronal activity in the sensory-motor cortex lasting throughout the stimulation, and a change in the firing rate related to spindles of the response. Adoption of the thalamic stimulation rhythm by cortical neurons could be observed either during the whole period of stimulation or in relation to phases of the recruiting response. These factors of influence of unspecific thalamic structures on neuronal activity in the hemispheres during recruiting response are relatively independent of each other. In contrast with data obtained in anesthetized animals, in chronic conditions unspecific structures exert heterogeneous mainly inhibitory influence on the spike activity of cortical neurons.


[The influence of oxytocin on the membrane potentials of mammary gland secretory cells].

The effect of oxytocine and its combinations with the acetylcholine, adrenaline, atropin, and prozerin on the level of membrane potential of mammary secretory cells was studied. Short oxytocine application (5-10(-4) ME/ml) evoked hyperpolarization of the contraluminal membrane (12+/-1 mV for 54+/-5 sec). Their response was increased by joint application of acetylcholine or proserine and decreased by preliminary application of atropin. Adrenaline increased or decreased the amplitude of the membrane potential's changes and considerably reduced the duration of the oxytocine reaction. Existence of oxytocine receptors was supposed on the membrane of secretory cells, as well as the possibility of acetylcholine and adrenaline effects on the structure of oxytocine receptors or the interaction of adreno--and cholinergic receptors with the oxytocine those.


[Cutaneous leishmaniasis: ultrastructural study (author's transl)].

While in general agreement with previous searchers, the authors direct their attention at peculiar or unknown structures such as: a huge phagosome sometimes loaded with a paracristalline rod; an occasional set of parallel microtubules along the reservoir; eventual duplication of the blepharoplast and even of the flagellum. There appear to be some 90 microtubules in the hyaloplasm of tleishmania tropica.


Effects of 25-hydroxycholesterol and aminoglutethimide in isolated rat adrenal cells. A model for congenital lipoid adrenal hyperplasia?

The production of corticosterone from 25-hydroxycholesterol by isolated rat adrenal cells is inhibited by aminoglutethimide phosphate (AGI); half-maximal inhibition is obtained at ca. 10 muM. AGI also inhibits ACTH-stimulated steroid production from endogeneous substrates; here half-maximal inhibition is obtained with ca. 40 muM AGI. In the presence of ACTH + AGI, 25-hydroxycholesterol causes additive inhibition. This effect of 25-hydroxycholesterol is dose-dependent. ACTH-stimulated steroid production from endogeneous substrates is partially inhibited by 5-cholene-3 beta,24-diol. These results may just reflect substrate competition for the side-chain cleaving system or may be due to some seocndary toxic effect on the cells.


Rapid interaction of prolactin with mouse mammary gland explants.

Exposure of mouse mammary gland explants to prolactin at 0 degrees C, for periods as brief as 10 seconds, caused a stimulation of labeled uridine incorporation into RNA during a subsequent incubation for 4 h at 37 degrees C. Furthermore, a 2-h wash of the prolactin-exposed explants in media at 0 degrees C did not attenuate the hormonal effect. A similar exposure of explants to insulin, followed by a 2-h wash at 0 degrees C, caused the abolition of the insulin stimulation of labeled uridine incorporation into RNA. The results suggest that there is a rapid and relatively stable interaction of prolactin with the mammary gland, while the interaction of insulin with this tissue would appear to be less stable.


FSH stimulation of DNA synthesis in Sertoli cells in culture.

The incorporation of [2H]thymidine into nuclear DNA was investigated in cultured Sertoli cells prepared from testes of 20-day-old rats. Addition of follicle-stimulating hormone (FSH) or dibutyryl cyclic, 3',5'-adenosine monophosphate (DBCAMP) to the culture medium greatly increased incorporation, expressed either as total amounts of [3H]thymidine incorporated per mug DNA or as the percentage of Sertoli cells with labeled nuclear DNA. No stimulation was observed in cells cultured in the presence of testosterone, insulin or cyclic 3',5'-GMP (cGMP). Light and electron microscopic autoradiographic analysis was employed to establish the identity of Sertoli cells having labeled nuclear DNA. Contaminating spermatogonia, which also took up labeled [3H]thymidine, were excluded from cell counts. In addition, Sertoli cells prepared from testes of irradiated 20-day-old germinal cell depleted rats were also observed to incorporate more [3H]thymidine into nuclear DNA when cultured in a chemically defined medium in the presence of FSH. DNA synthesis was abolished by prior treatment of cells with cytosine arabinoside. In separate experiments, the incorporation of [3H]thymidine into DNA of peritubular myoid cells was shown to be independent of FSH or dbcAMP.


Characterization of the protein kinases in a transplantable islet cell tumor of the Syrian hamster.

The protein kinase activities of a transplantable, insulin-producing hamster islet cell tumor were characterized using gel filtration, sucrose density gradient centrifugation and acrylamide gel electrophoresis. The post-microsomal supernatant fluid contains 70-80% of the protein kinase activity present in crude homogenates. A cAMP-dependent protein kinase, PK I (Mr 170,000), represents 25% of the soluble protein kinase activity assayed with protamine as substrate. It dissociates in the presence of cAMP into a cAMP-binding protein, R2 (Mr 90,000) and a catalytic subunit C (Mr 33,000). The dissociation induced by cAMP seems to be facilitated by the addition of Mg2+ and ATP. The regulatory subunit, R2, changes its gel filtration pattern in the presence of 0.5 M NaCl suggesting dissociation into a smaller subunit, R1 (Mr 44,000). By analogy with purified beef heart protein kinase (Erlichman et al., 1973) and skeletal muscle protein kinase, PK I. The presence in crude homogenates of a free cAMP-binding protein indistinguishable from the R2 derived by dissociation of PK I, suggests that PK I is partially dissociated in vivo. A cAMP-independent (casein) kinase (Mr 210,000) elutes with PK I on columns of Sepharose 6B. Another cAMP-independent protein kinase, PK II (Mr 88,000), is the predominatn form of soluble protein kinase accounting for approximately 75% of the soluble protein kinase activity detected using protaimine as substrate. This cAMP-independent protein kinase changes its gel filtration pattern in the presence of 0.5 M NaCl giving rise to a form which appears to have the same Mr (33,000) as the catalytic subunit of PK I. Studies comparing the catalytic subunit C of PK I with PK II and its salt-induced smaller molecular form demonstrate facile association of C with the cAMP-binding protein of purified bovine heart protein kinase to yield a hybrid holoenzyme, whereas PK II and its smaller form fail to recombine in this fashion. The 33,000 dalton forms derived from PK I (by cAMP) and PK II (by salt) also show different substrate specificities. It would appear, therefore, that pK II is a cAMP-independent protein kinase unrelated to PK I.


LH- and FSH-stimulating of adenylate cyclase in seminiferous tubules from young rats: functional FSH and LH receptors unmasked by homogenization.

LH- and FSH-sensitive adenylate cyclase activity was present in homogenates of whole testis tissue as well as in microdissected seminiferous tubules derived from young rats. In homogenates of seminiferous tubules a single adenylate cyclase appears to interact with both LH and FSH through separate hormone-specific receptors. Disruption of testis tissue by homogenization exposes functional FSH and LH receptors which are inaccessible to the hormones in intact cell preparations. These results indicate that in certain seminiferous tubule cell types only a fraction of the total functional receptors present is accessible to the cell surface for interaction with hormone.


Gastric antisecretory and antiulcer properties of PGE2, 15-methyl PGE2, and 16, 16-dimethyl PGE2. Intravenous, oral and intrajejunal administration.

15-Methyl PGE2 and 16,16-dimethyl PGE2 were found (1) to be 40 and 100 times, respectively, more potent than PGE2 after intravenous administration in inhibiting histamine-stimulated gastric secretion in dogs with a denervated (Heidenhain) gastric pouch, (2) to be active orally and intrajejunally, whereas PGE2 was inactive, and (3) to exert antisecretory activity for longer duration than PGE2. 16,16-Dimethyl PGE2 was about 2.5 times more potent than 15-methyl PGE2. Volume, acid concentration, and output, and pepsin output (but not concentration) were reduced in a dose-dependent manner. In the rat, 16,16-dimethyl PGE2 also inhibited gastric secretion and prevented the formation of ulcers produced by various methods: gastric ulcers (Shay, and steroid induced) and duodenal ulcers (secretogogue induced). In this species, 1l816-dimethyl PGE2 was 2 to 50 times more potent than PGE2, depending on the endpoint, and was active orally. These prostaglandins appear to inhibit gastric acid secretion by acting directly on the parietal cells, and making these unresponsive to most stimulants. Vomiting was a side effect of the prostaglandin analogues in the dog, but almost exclusively when these were given orally. After intravenous or intrajejunal administration at doses inhibiting gastric secretion by 80%, vomiting was seen only once. These results suggest that 15-methyl PGE2 and 16,16-dimethyl PGE2 may be of value in the treatment of peptic ulcer.


Fulminant herpes simplex hepatitis in an adult: report of a case in renal transplant recipient.

A case of disseminated herpes simplex infection is reported in a 31-year-old renal transplant recipient. The patient presented with a unique clinical syndrome: high fever, severe sore throat with buccal and pharyngeal ulcerations, fulminant hepatitis, thrombocytopenia, and leukopenia. The patient died from hepatic failure, disseminated intravascular coagulopathy, and upper gastrointestinal bleeding. The diagnosis was made by positive herpes simplex virus culture from the throat, and was confirmed at autopsy by typical Cowdry's type A intranuclear inclusions in hepatocytes with positive herpes simplex virus culture from the liver. Review of the literature reveals that other reported cases have had very similar clinical findings, making disseminated herpes simplex infection with fulminant hepatitis a recognizable syndrome.


[Seroepidemiological studies related to the association of genital herpes to cervical cancer (author's transl)].

Blood sera of 290 patients were examined for the presence of antibodies against herpes simplex virus type 1 and type 2 (HSV-1 and HSV-2): 1. lues - seronegative individuals, 2. patients with specific lues antibodies, and 3. patients with so-called carcinoma in situ and invasive carcinoma of the uterine cervix. The group of syphilis-seropositive individuals served as control for the group with cervical lesions, because in both groups certain socioepidemiologic factors (promiscuity, early sexual intercourse) appear to be prevalent. The average value for HSV-2-antibodies was significantly higher in patients with cervical lesions than that in lues-seropositive individuals. Also, the average value was higher in the lues-seropositive group than that in the lues-seronegative group. The frequency of HSV-2-antibodies was highest in the group of patients with so-called carcinoma in situ and invasive carcinoma, less in the lues-seropositive group, and even more less in the lues-seronegative group. These results make it unlikely that the connection between cervical carcinoma and HSV-2-infection is a common or "nonsense"-correlation. They suggest a more narrow correlation between both diseases, but leave open whether it consists of either a causal relationship between herpes infection and the development of cervical carcinoma, or a predilection for herpes infection in the cervical region by the carcinomatous tissue.


Mapping and gene conversion studies with the structural gene for iso-1-cytochrome C in yeast.

We have investigated the order of the four genes cyc1, rad7, SUP4, and cdc8 which form a tightly linked cluster on the right arm of chromosome X in the yeast Saccharomyces cerevisiae. Crossing over and coconversion data from tetrad analysis established the gene order to be centromere-cyc1-rad7-SUP4. Also cdc8 appeared to be distal to SUP4 on the basis of crossovers that were associated with conversion of SUP4. The frequencies of recombination and the occurrence of coconversions suggest that these four genes are contiguous or at least nearly so. Gene-conversion frequencies for several cyc1 alleles were studied, including cyc1-1, a deletion of the whole gene that extends into the rad7 locus. The cyc1-1 deletion was found to be capable of conversion, though at a frequency some fivefold less than the other alleles studied, and both 3:1 and 1:3 events were detected. In general 1:3 and 3:1 conversion events were equally frequent at all loci studied, and approximately 50% of conversions were accompanied by reciprocal recombination for flanking markers. The orientation of the cyc1 gene could not be clearly deduced from the behavior of the distal marker SUP4 in wild-type recombinants that arose from diploids heteroallelic for cyc1 mutations.


[3 new allotypes of mink blood serum alpha2-lipoproteins--Lpm 6, Lpm 7, Lpm 8].

Three new allotypes were discovered in mink sera by means of isoimmunization. Based on the results of identifications, they were referred to the Lpm system of serum alpha2-lipoprotein. They were designated as Lpm 6, Lpm 7 and Lpm 8. The allotyping of serum samples from 342 minks made possible to establish a relation between Lpm 7 and Lpm 8 using the chi2 method; it was also found that these two allotypes related to each of the first five Lpm as well as to their phenotypes, which was described earlier. There was a significant dependence of the expression of Lpm 6 on Lpm 5 indicating a genetic relation between Lpm 6 and other allotypes. The detection of Lpm 6, 7 and 8, which are the most likely under the control of the same gene (or a sustem of genes) as Lpm 1, 2, 3 and 5, is an evidence for the complex structure of the Lpm locus.


Early stimulation of human chorionic gonadotropin secretion by dibutyryl cyclic AMP and theophylline in human malignant trophoblast cells in vitro: inhibition by actinomycin D, alpha-amanitin, and cordycepin.

Human malignant trophoblast cells (BeWo line) in culture were employed to investigate the early stimulation of human chorionic gonadotropin (hCG) secretion by 1 mM dibutyryl cyclic AMP and 1 mM theophylline (dbT). The earliest increase in secreted immunoreactive hCG occurred at 3 1/2 h following addition of dbT, and was preceded by an increase in intracellular hCG. These results suggested that dbT stimulated hCG synthesis, rather than release. Addition of either 0.062 mug/ml actinomycin D or 100 mug/ml cordycepin along with dbT, or 3 mug/ml alpha-amanitin 9 h prior to addition of dbT, prevented the increase in hCG secretion at 3 1/2-8 h. It was concluded that synthesis of RNA (possibly messenger RNA) is required for the early stimulation of hCG secretion by dbT.


[The diagnosis of carcinoma of the breast in relation to growth rate (author's transl)].

The doubling time (DT) was estimated quantitatively for 16 carcinomas of the breast according to the method described by Collins and co-workers in 1956. This is based on the concept of constant and exponential growth. Observation interval for these mammographically confirmed tumors was between 83 and 1,034 days. The DT was calculated to vary from 45 to 260 days; in order to reach a diameter of 1 cm. after 30 divisions would require a period of 3.7 to 21.4 years. Mammography frequently demonstrates small, clinically occult, tumours. Axillary lymph node metastases are relatively rare from small tumours; growth rate of 70% of breast carcinomas in such that an annual clinical and radiological check-up will prove to be the best means of reducing mortality from carcinoma of the breast. The risk inherent in the radiation resulting from annual mammography is acceptable in women over 35 years. This leaves the problem of rapidly growing carcinomas which would escape early diagnosis by early examinations. Half-yearly examinations of women in high risk groups (1. Previous mastectomy for carcinoma, 2. Biopsy-proven mastopathy with atypical proliferation) comprising about 30% of carcinomas with a short doubling time would appear to be reasonable.


[A comparison of radiology and scanning with 99mTc-pyrophosphate in 125 patients with tumours (author's transl)].

Various portions of the skeleton were examined in 125 patients both radiologically and by scanning, using 99mTc-pyrophosphate. The results are compared, particular attention being paid to errors in diagnosis. The figures show that scans represent a valuable additional form of examination for detecting tumours which have not been found radiologically. The importance of serial observations is discussed.


[The distribution of Sn2+ after application as a complex (author's transl)].

On animal experiments was shown that Sn2+ after i.v. application with 113Sn as radioactiv indicator was deposited in the skeleton. Experiments were done with citrat-, tartrat-, pyrophosphat-, phosphat- and Sn2+/HCl-complexes. The importance of the results for the use of Sn2+-solutions in Nuclear Medicine for men are discussed.


[Klippel-Trenaunay's syndrome (author's transl)].

The authors have observed seven cases of this syndrome, and together with a review of the literature, they discuss the various facets which constitute this disease. Particular attention is paid to the findings in the vessels of the extremities on phlebography and arteriography, and the authors conclude that these examinations are vital in the diagnosis of this syndrome.


[Stretched veins, a new, indirect sign of malignant breast tumours (author's transl)].

The diagnosis of malignant breast tumours depends on direct and indirect radiological signs. Amongst the latter, considerable emphasis has been placed in the literature on the finding of dilated veins. An analysis of the literature and of nearly 1,500 examinations has shown, however that this sign is not as reliable as has been thought. A new sign has been noticed, which has so far not been emphasised in the literature. This is stretching of the veins. It is not a direct sign, but as an indirect sign it is a more reliable indication of malignancy than dilatation of veins. Stretching of veins may also occur in benign conditions, but it is twice as frequent in the presence of malignant tumours. It is therefore considered that the presence of stretched veins is an indirect sign of malignant breast tumours.


[Spread of acute haemorrhagic conjunctivitis virus in virgin soil (Hokkaido, Japan): observation with serological surveys (author's transl)].

Acute haemorrhagic conjunctivitis (AHC), a novel disease caused with a new type of enterovirus, occurred for the first time in Sapporo City and other areas of Hokkaido in October of 1971. During first 4 months, 1,000 or more cases of AHC were reported in Sapporo and 2,300 cases in other areas. Thereafter, there are no epidemic of AHC in any areas, though there are sporadic cases up to date. To know the spread of exogenous agent, AHC virus, in Hokkaido, the neutralizing antibody was measured with sera of healthy adults resident in 12 separated districts of Hokkaido in 1973 and 1974. It was revealed that the positive reactors to AHC virus were found in 5 to 35% in all districts but one in 1974, and AHC virus was distributed in almost all the areas of Hokkaido within a period of two and half years after invasion. In addition, a sequential sero-survey to detect the spread of AHC virus has been carried out in Sapporo. A rate of neutralizing antibody positives in healthy adults did not show a clear increase on sera withdrawn one year after epidemic of AHC in comparison with those before epidemic, but show a remarkable increase on sera withdrawn later in spite of non-epidemic. An increase of antibody positive rate in non-epidemic period was also obtained from sera of healthy adults resident in districts other than Sapporo. These findings made doubt the changes in transmission on mode and/or in clinical pictures, and the viral mutation in biological character, such as temperature for propagation, may be supposed.


The plasmatic 11-hydroxy-corticosteroid response in the human to a synthetic octadecapeptide corticotrophin (C-41795-Ba).

The adrenal response as measured by the plasmatic changes in 11-hydroxy-corticosteroid (11-OHCS) concentration following the administration of a synthetic octadecapeptide (C-41795-Ba) was compared in 17 normal young males with that obtained with the commercially available tetracosactrin. C-41795-Ba polypeptide shows a higher potency than tetracosactrin. Its effects are longer lasting when administered intravenously which indicates a significant local inactivation of the polypeptide.


Binding of somatomedins and insulin to plasma membranes prepared from rat and monkey tissue.

Particulate membranes prepared from a variety of monkey and rat tissues were shown to have specific binding sites for somatomedin A. Binding to talc on the other hand showed no specificity, and the sensitivity was less. The membrane-bound somatomedin A was displaced by somatomedin A in concentrations between 0.01 and 5 U/ml. Of the other hormones tested only insulin in high concentrations could interfere with the binding of labelled somatomedin A to tissue membranes. Membranes prepared from a number of rat and monkey tissues also contained binding sites for insulin. Somatomedin A could interfere with the binding of labelled insulin. No binding was observed for somatomedin B in the tissues studied.


Impairment and restoration of response to TSH in dog thyroid slices after treatment with phospholipase A and lubrol PX.

Incubation of dog thyroid slices with phospholipase A (10-40 U/Ml) or Lubrol PX (0.08-0.4%) caused a diminution in the subsequent TSH effect on the tissue cyclic AMP level and glucose oxidation. The same treatment had no effect on the basal level of these parameters. When the phospholipase A or Lubrol PX-treated slices were rinsed intensively with a Krebs-Ringer bicarbonate buffer and then incubated at 37degreesC in the same buffer for a further 1 to 3 hours, responsiveness to TSH recovered progressively reaching almost completely that of the control slices. Again, these procedures were without any significant effect on the responsiveness of the control slices. The above results together with those reported previously suggest strongly that phospholipids are an essential component of the plasma membrane system by which TSH stimulates adenylate cyclase activity. In addition, these essential lipids in the membrane appear to be renewed rather efficiently in this tissue, thus securing the functional integrity of the thyroid in the face of various deleterious situations.


The effect of ACTH administration on the phagocytic and bactericidal activities of human polymorphonuclear leukocytes.

Metabolic and bactericial activities of leukocytes obtained from 5 normal volunteers receiving 1 mg of synthetic B1-24 ACTH intra-muscularly daily for 7 days, were studied. Bactericidal activity and phagocytosis induced hexosemonophosphate shunt activity of leukocytes were found to be depressed following ACTH administration. However, glycolytic activity, which provides the necessary energy for particle uptake by leukocytes, was not altered. These findings indicate that a moderately prolonged exposure to elevated levels of plasma cortisol does not affect the phagocytic activity of leukocytes (as indicated by glycolytic activity) but significantly impairs their ability to destroy the ingested bacteria. It is suggested that the impaired bactericidal activity of leukocytes reported in children suffering from protein-calorie malnutrition may partly be due to elevated plasma cortisol levels seen in them.


Synthesis of 9-O-acetyl- and 4,9-di-O-acetyl derivatives of the methyl ester of N-acetyl-beta-D-neuraminic acid methylglycoside. Their use as models in periodate oxidation studies.

Reaction of the methyl ester of N-acetyl-beta-D-neuraminic acid methyl glycoside with N-acetylimidazole yielded the corresponding 9-O-acetyl- and 4,9-di-O-acetyl derivatives. The structures of these compounds were confirmed by mass spectrometry and both 1H and 13C NMR spectroscopy. The compounds served as model substances in a comparative study of the rate of periodate oxidation of unsubstituted and of 9-O-acetylated-N-acetyl-neuraminic acids. This reaction was strongly hampered by the presence of the 9-O-acetyl group. The low molar absorbancy coefficient of N-acetyl-9-O-acetylneuraminic acid in the periodic acid/thiobarbituric acid assay can be explained by this retardation.


Quantitative determination of poly(adenosine diphosphate ribose) in different hepatic tissues by an isotope dilution procedure.

A procedure has been developed for the quantitation of poly(ADP-ribose) in intact tissues. It is based on the dilution of added [3H]poly(ADP-ribose) by the endogenous polymer. 5 - 6 nanomoles protein-bound ADP-ribose per mg DNA were found in adult and neonatal rat liver, while Zajdela hepatoma cells had significantly lower values. A comparison with mono(ADP-ribose) residues in adult rat liver revealed similar levels of monomeric and polymeric ADP-ribose residues. This means that far more proteins (or acceptor sites on proteins) must be occupied by single ADP-ribose residues than by oligo or poly(ADP-ribose) chains. While the poly(ADP-ribose) levels of the different tissues do not correlate with the corresponding proliferation rates, the amount of mono(ADP-ribose) does show a certain Correlation, being low in rapidly growing tissues.


Isoenzymes of serum N-acetyl-beta-D-glucosaminidase in the I cell disease heterozygote.

Serum N-acetyl-beta-D-hexosaminidase is compared quantitatively and qualitatively in 14 obligate heterozygotes for the mutant gene causing I cell disease (ICD) or mucolipidosis II and in 31 normal controls. The average specific activity in either group is significantly different but reliable heterozygote detection cannot be achieved because of some overlapping of the ranges of individual results. Fractionation of the enzyme either by DEAE cellulose column chromatography, or by heat inactivation, yields a typical average result for each genotype. Also, mere expression of the various components as percentages of the total activity is not useful for certain identification of the ICD heterozygote. There is considerable overlapping of the percents hexosaminidase I1 and A in both groups of sera. If enzymatic hydrolysis by any component is expressed as a partial activity, a much better though not absolute distinction between the ICD heterozygote and the normal control becomes possible. Only the latter way of expression of hexosaminidase results makes distinction between the ICD heterozygote and the Tay-Sachs heterozygote very probable.


Carcinoma of the esophagus in Africans: establishment of a continuously growing cell line from a tumor specimen.

Biopsy material, from a patient with an esophageal carcinoma of a type occurring relatively frequently in Africans in some regions of Southern Africa, has been adapted to cell culture conditions and has developed into a continuously growing tumor cell line. The cells have a squamous pattern of growth which they maintain, without piling up, even in heavily confluent cultures.


Identification of cholesterol in the receptor site for rickettsiae on sheep erythrocyte membranes.

Cholesterol was identified as an essential component of the receptor on the sheep erythrocyte to which Rickettsia prowazeki adsorbs before lysing the cell. Erythrocytes or ghosts, derived by hypotonic lysis, were treated with proteolytic enzymes, sialidase, sulfhydryl reagents, and periodate without affecting their ability to adsorb rickettsiae. Lipid extracts of ghosts and erythrocytes, on the other hand, contained receptor activity. Fractionation of the lipid extracts by silicic acid column chromatogrphy resulted in the isolation of receptor activity in a neutral lipid fraction. The lipid fractions demonstrated receptor acitity at 34 C but not at 0 C. These properties are also characteristic of the receptor activity with erythrocytes and ghosts. Cholesterol, co-lyophilized with palmitic acid, was found to possess receptor activity. Palmitic acid alone, cholesterol-lecithin, cholestane-palmitic acid, and various phospholipids and glycolipids had no receptor activity. Ghosts treated with amphotericin B or digitonin, compounds that bind to cholesterol in the membrane, lost their ability to adsorb rickettsiae.


Role of Ca2+ and Mg2+ in some human neutrophil functions as indicated by ionophore A23187.

Studies with the divalent cation ionophore A23187 suggest that both Ca2+ and Mg2+ ion influx play a role in human peripheral blood neutrophil function. Degranulation of neutrophils occurred at ionophore concentrations of 10(-5) M and was Ca2+ but not Mg2+ dependent. Modulation of neutrophil chemotaxis was enhanced optimally by 10(-7) M ionophore and was both Ca2+ and Mg2+ dependent. Concentrations of ionophore as low as 10(-12) M seemed to sensitize the cells to a concentration of phorbol myristate acetate which by itself was not chemotactic. These findings also indicate that factors other than Ca2+ or increases in cyclic nucleotides are important to initiation and modulation of neutrophil function.


Increased cyclic adenosine 3',5'-monophosphate content in guinea pig ileum after exposure to Staphylococcus aureus delta-toxin.

To compare Staphylococcus aureus delta-toxin with cholera toxin, which is known to increase cellular cyclic adenosine 3',5'-monophosphate (cAMP), studies were undertaken to determine the effect of delta-toxin on the cAMP content of guinea pig ileum maintained in vitro. Concentrations of delta-toxin as low as 0.40 mug/ml increased cAMP levels in guinea pig ileum after 2 h of incubation. Histological damage was seen in ileum exposed for 2 h to delta-toxin concentrations of 100 mug/ml. As little as 3 mug of delta-toxin increased vascular permeability in guinea pig skin. Permeability changes became evident within 5 min and were maximal within 6 h, whereas those produced by cholera toxin required 24 h to become maximal. Benadryl did not interfere with the ability of these toxins to alter vascular permeability. Purified egg lecithin reduced the effectiveness of delta-toxin in the skin but did not inhibit cholera toxin. Delta-toxin in concentrations as low as 0.1 mug/ml caused dislodgement of HeLa cells in tissue cultures. Therefore, delta-toxin appears unique in being the only bacterial toxin, currently known to alter water absorption in the ileum, that is capable of both increasing cAMP levels and being cytotoxic. These findings suggest a possible role for delta-toxin in the pathogenesis of staphylococcal enteritis.


Variability in the structural polypeptides of herpes simplex virus 1 strains: potential application in molecular epidemiology.

This paper reports on the variability of structural polypeptides of 53 strains of herpes simplex virus 1 isolated from Italy, Uganda, South Africa, and various locations in the United States. Most strains were passaged a limited number of times at low multiplicity outside the human host; a few strains were characterized by numberous passages at variable multiplicities in cell culture and experimental animals. The acrylamide gel electrophoresis of polypeptides from purified virions revealed seven variable polypeptides. Virion polypeptides (VP) 7, 11, 13, 14, 15.2 and 23 were present in at least two isotypic forms characterized by fast and slow electrophoretic mobilities. VP8 could not be detected in three strains. In addition, VP13, 15.2, and 23 in some strains were either absent or comigrated with other polypeptides. A variety of tests showed that the variability in electrophoretic mobility of polypeptides was reproducible and could not be attributed to artifacts of purification, solubilization, or electrophoresis. Attempts to classify the strains on the basis of electrophoretic mobility of five or all seven variable polypeptides yielded 14 and 19 groups, respectively. The bulk of the strains (41 to 53) fell into six groups. Not all possible permutations of variable polypeptides were observed. Comparison of early and late passages of laboratory strains showed that in the few instances tested the variability could not be attributed to the propagation of the virus outside the human host. Clustering of strains on the basis of country of origin was not observed. Some clustering of isolates on the basis of site of isolation was observed, and the data do suggest that further analyses of isolates for evidence of a correlation between the site of localization on the human body and the structural polypeptides might be useful. Electrophoretic characterization of structural polypeptides has the potential of becoming a powerful tool for epidemiological studies of herpes simplex virus infections.


Characteristics of cells present in peritoneal fluids of mice injected intraperitoneally with Bordetella pertussis.

Peritoneal fluids obtained from mice after the intraperitoneal administration of Bordetella pertussis vaccine, heated vaccine, an extract of the organisms, killed Escherichia coli, or thioglycolate medium were examined in terms of total cells and percentage that adhered to glass cover slips during 2-h incubation period. All these substances were found to increase the number of leukocytes in peritoneal fluid within 1 to 2 days after the injection. This increase appeared to be due to an influx of macrophages and polymorphonuclear leukocytes with relative proportions at a given time dependent upon the material involved in the induction of the response. The initial increases after pertussis vaccine seemed to be due mainly to an influx of monomuclear cells, whereas with E. coli neutrophils constituted the major portion of the cell population. The percentage of peritoneal cells that attached to glass was also found to be markedly reduced in preparations obtained from mice after the injection of B. pertussis or E. coli. There appeared to be differences in persistence of this phenomenon, with preparations containing the histamine-sensitizing factor being the most active in affecting adherence properties. Thus these data would suggest that the action of B. pertussis on macrophages (or precursors) and neutrophils is not expressed in terms of suppression of emigration properties, as has been reported by others for lymphocytes, but is manifested in the alteration of glass-adherence characteristics. Within experimental limitations, it is believed that macrophages are possibly more involved in terms of altered function than are the polymorphonuclear cells.


Effect of in vitro adaptation of Marek's disease virus on pock induction on the chorioallantoic membrane of embryonated chicken eggs.

Cell-associated preparations of several isolates of Marek's disease virus produced more pocks on the chorioallantoic membrane of embryonated chicken eggs than plaques in duck embryo fibroblasts, thus indicating that lesion response in eggs was more sensitive than cytopathic response in duck embryo fibroblasts for assaying low-passage Marek's disease virus. Adaptation of the virus to cell cultures by serial passages, however, substantially reduced its pock response so that the titer ratio (plaque-forming units in duck embryo fibroblasts/pock-forming units in eggs) of cell culture-adapted Marek's disease virus was 1 or higher. The decreased pock response could not be attributed to selection of preexisting virus variant(s) with low affinity for chorioallantoic membrane because cloned Marek's disease virus had a good pock response at low cell culture passage levels, but this response decreased as the virus was attenuated by serial cell culture passage.


Inhibition of lipopolysaccharide-initiated activation of serum complement by polymyxin B.

Treatment of bacterial lipopolysaccharides (endotoxins) isolated from Salmonella minnesota R595 or purified lipid A with the antibiotic polymyxin B has been demonstrated to abrogate the ability of these molecules to activate serum complement. The inhibition is a function of the ratio of polymyxin B to lipopolysaccharide.


Epstein-Barr virus-specific IgA serum antibodies as an outstanding feature of nasopharyngeal carcinoma.

Stimulated by a report on elevated IgA levels in nasopharyngeal carcinoma (NPC), we tested a total of 372 sera from patients with NPC, other carcinomas of head and neck or elsewhere, Burkitt's lymphoma (BL), infectious mononucleosis (IM) or healthy controls. The sera were titrated in indirect immunofluorescence tests for IgA antibodies to Epstein-Barr virus (EBV) capsid antigen (VCA) and to the diffuse (D) or restricted (R) components of the EBV-induced early antigen (EA) complex. The results proved NPC to be outstanding in that prior to therapy 93% of the patients tested revealed IgA antibodies to VCA and 73% to D, often at high titers which occasionally matched the corresponding IgG antibody levels. The EBV-specific IgA titers increased from stages I or II to stages III or IV; i.e. with the total tumor burden. Conversely, many of the NPC patients examined 2-6 years after initial therapy had only low levels of EBV-specific IgA or none at all, and the majority of those with high titers were known to have residual or recurrent disease. In contrast to untreated NPC patients, less than 5% of 73 patients with other carcinomas or of 76 healthy donors revealed VCA-specific IgA and even fewer EA-specific IgA; only 28% and 4% of 54 BL patients tested at admission had IgA antibodies to VCA and R, respectively, and 38% and 3% of 37 IM patients showed transient VCA- or D-specific IgA responses, all at generally low titers. While sera from untreated NPC patients often contained IgA antibodies also to herpes simplex type 1 virus, their incidence and range of low titers were similar to those obtained with sera from patients with other carcinomas or from healthy donors. It thus appears that the elevated IgA levels in NPC might be due to EBV-specific antibodies. Possible reasons for this unique response in NPC have been discussed.