[Quinolozidinylalkylamines with antihypertensive activity].

Since lupinylamine [(I); R = H] exhibits hypotensive activity, mainly due to ganglionic blocking properties, and it is known that a high degree of steric hindrance around the basic function of other ganglioplegic amines is of paramount importance for optimal activity, several guinolizidine derivatives were prepared. They differ in the length of the alkyl chain connected to the ring and in the position of the amino group along the chain. Some N-substituted derivatives of 2-quinolizidin-1'alpha-yl-ethylamine (II) together with O-lupinylhydroxylamine and 2-quinolizidin-1'beta-yl-ethylamine, respectively isosteric and epimeric to it, were also prepared. When administered orally to spontaneously hypertensive rats, the amines (II), (III) and (XI) produced high and long-lasting antihypertensive activity, while the remaining compounds so far tested were inactive or had only modest effect on blood pressure. Compared with alpha-methyl-DOPA, amine (II) appears to be approximately two to three times as potent. The antihypertensive activity of (II) appears to be linked to glanglionic blocking properties, since this amine proved 1,2 times as potent as mecamylamine in the inhibition of cat nictitating membrane response to stimulation of the preganglionic sympathetic nerve.


Use of iminodiacetic acid for the synthesis of piperazinyl peptides related to leu-enkephalin. II.

Piperazinyl peptides related to Leu-enkephalin were obtained using as intermediates condensation products with iminodiacetic acid. The products are devoid of analgesic activity.


Relationships between growth hormone secretion and microangiophaty in subjects with prediabetes.

This study was aimed at evaluating the relationship between growth hormone secretion and the presence of functional microangiopathy in prediabetes. The results here in presented clearly indicate that an exaggerated and abnormal secretion of this hormone was a constant feature of those prediabetics with functional angiopathy.


Drug-protein interaction: plasma protein binding of furocoumarins.

The binding of six furocoumarins (angelicin, psoralen, 8-methoxypsoralen, 5-methoxypsoralen, 8-methylpsoralen, 4,5'8-trimethylpsoralen) to human serum and human serum albumin was studied by equilibrium dialysis using tritium labelled compounds. The results indicated that in serum all furocoumarins are bound mostly by albumin, the extent of binding being related to the structure of the furocoumarins; at any rate, high values of the bound drug, ranging from 84 to 97% were observed. The percentage of binding is strictly related to the water solubility of the compounds. A limited number of binding sites, n = 1-2.4, were detected in the albumin molecule, indicating a high specificity in the binding process. The association constants of the furocoumarins to albumin. Ka, ranged from 1.2 X 10(4) M-1 (8-methoxypsoralen) to 1.9 X 10(5) M-1 (4,5'8-trimethylpsoralen).


Menstrual history of young women exposed in utero to diethylstilbestrol.

The menstrual histories of 218 patients exposed in utero to diethylstilbestrol (DES) were compared with those of 158 control subjects. No significant differences were revealed in the complaint of menstrual irregularities at either the initial or follow-up examination. Age at menarche was the same in both groups, and was the same as found in the United States as a whole. These findings differ from the report of a controlled trial in Chicago which suggests that there is a specific pattern of oligomenorrhea associated with DES exposure.


The role of counseling in infertility.

Sixty-two couples with infertility received infertility counseling along with the infertility investigation and treatment. At the end of the study a questionnaire was sent to all patients in order to evaluate the incidence of psychologic symptoms associated with their infertility and their response to counseling. The study confirmed the presence of a high incidence of emotional symptomatology in the self-selected patients. It showed that male partners are also affected, but less so. Finally, it has demonstrated that infertility counseling serves to enhance the quality of life in many patients who have become the victims of their "infertility crises."


Therapeutic donor insemination: results and causes of nonfertilization.

The results of therapeutic donor insemination (AID) in 108 women are presented. Of the 108 women, 65 (60%) achieved a pregnancy. Eighty-five per cent of the pregnancies occurred with six AIDs or less, and 80% resulted in live births. The reasons why 43 women did not conceive are analyzed. The most common cause of failure was discontinuing the AID program. Of the original 108 women, 34 (32%) had dropped out of the program by the time they had received 6 AIDs or less. Reproductive abnormalities in the wife, and irregular ovulatory patterns, were the next most common causes of failure, and were often associated with discontinuation of the program. Advice regarding 2 days of sexual abstinence prior to AID was considered to improve the pregnancy rate. Fresh semen was shown to be superior to frozen samples in achieving pregnancies.


Induction of ovulation in women with hyperprolactinemic amenorrhea using clomiphene and human chorionic gonadotropin of bromocriptine.

Clomiphene citrate (Clomid), when given alone, is generally considered ineffective in inducing ovulation in women with hyperprolactinemia. This study reports the treatment of 29 infertile women with hyperprolactinemic amenorrhea. Twenty-one patients (eighteen of whom had previously had no ovulation response to Clomid alone) were treated with a combined regimen of Clomid (100 to 200 mg/day for 5 days) and two injections of 5000 IU of human chorionic gonadotropin (HCG), the first 8 to 10 days after Clomid withdrawal and a second injection 1 week later. Basal body temperature charts, conception, and/or plasma progesterone measurements showed that 19 patients ovulated (90%). There were 17 pregnancies in 12 of 21 patients (57% pregnancy rate) with 15 single live births and two abortions. When bromocriptine (Parlodel) became available, a total of 22 patients (including 14 patients previously treated with Clomid/HCG, six of them successfully) with amenorrhea associated with hyperprolactinemia were treated with this drug with dosages varying from 2.5 mg to 15 mg/day. Ovulation was confirmed in 20 patients (90%). There were 17 pregnancies in 15 patients (68% pregnancy rate) with 15 single live births and two first-trimester abortions. In all, 21 of 29 patients (73%) achieved one or more pregnancies resulting in live births with one or both of the above treatments. It is concluded that a combined Clomid/HCG regimen can often be used as an effective alternative to bromocriptine therapy in the treatment of infertility associated with hyperprolactinemic amenorrhea.


Blood carbon dioxide tension changes during hysteroscopy.

During hysteroscopy the uterus may be distended with carbon dioxide (CO2), nitrous oxide (N2O), or Hyskon (a high molecular weight dextran). An initial study in 27 patients (group 1) using arterialized venous blood samples demonstrated rises in carbon dioxide tension (PCO2) when N2O was insufflated by using a laparoscopy insufflating device--a constant-pressure, variable-volume gas source. Cardiovascular collapse occurred in one patient in this group, most probably as a result of macropulmonary emboli of N2O. The rise in PCO2 is accounted for by an increase in physiologic dead space. In another 24 patients (group 2) the gaseous media were introduced by using a constant-volume, variable-pressure gas source; this resulted in minimal changes in arterial PCO2. The choice of whether a gaseous or liquid distending medium is used for hysteroscopy is governed by the state of the endometrium. If a gaseous medium is indicated, then CO2 is preferable to N2O and should be introduced with a constant-volume, variable-pressure gas source.


End-to-end tubal anastomosis using an absorbable stent.

Using absorbable suture as a stent, end-to-end anastomosis of previously ligated or cauterized fallopian tubes was performed in 15 cases. Luminal patency of at least one fallopian tube has been achieved in all cases, and thus far seven pregnancies have occurred in six cases. The technique for inserting the absorbable suture into the distal and proximal portions of the tube is fully described, and the importance of repeated hydrotubations to maintain tubal patency is emphasized.


An estimate of the fertility potential of the fractions of the split ejaculate in terms of the motile sperm count.

The motile sperm count is a simple index which is equivalent to the product of sperm density (millions of sperm per milliliter) and percentage of motility (motile sperm per 100 sperm x 0.01). Since other studies have demonstrated a positive correlation between this index and the pregnancy rate, it has been hypothesized that the motile sperm count could be useful for expressing the potential fertility of individual semen specimens. Collection of semen by the split ejaculate technique and subsequent artificial insemination homologous (AIH) with the best split fraction has been used for the treatment of oligospermia. We calculated the motile sperm count for whole semen and the first fraction of split ejaculates from 45 men who were studied for infertility. Subsequently, 15 couples agreed to AIH with the first fraction of the split ejaculate. The data indicate that the motile sperm count may be a reasonable index for comparing the potential fertility of the semen specimens and thereby estimating the chances of successful AIH.


Prolactin, fructose, and zinc levels found in human seminal plasma.

Fructose, zinc, and prolactin determinations were performed on the seminal plasma of normospermic (n = 30), oligospermic (n = 30), and azoospermic (n = 30) men to determine whether there was a correlation between any of these substances and seminal quality. None of these azoospermic men had congenital bilateral absence of the vasa, but in a separate group of such men (n = 10) these same studies were performed. Fructose was determined photometrically, zinc by atomic absorption, and prolactin by radioimmunoassay. Zinc and fructose levels in all three groups showed no statistical differences. Prolactin was found to be significantly higher in the seminal plasma of normospermic men than was found in the seminal plasma of oligospermic or azoospermic men. In men with congenital bilateral absence of the vasa and seminal vesicles, zinc was elevated, fructose was absent, and prolactin was markedly depressed.


Effects of antisera on human sperm penetration of zona-free hamster ova.

Human sperm, after treatment with a 10% concentration of rabbit or rhesus monkey normal sera and antisera, were evaluated for fertilizing potential by incidence of zona-free hamster ova penetrated by the sperm as evidenced by the presence of swollen sperm heads and male pronuclei. Compared with the basic medium alone, treating sperm with normal sera tended to increase the percentage of ova penetrated whereas antisera against sperm, sperm extract, and testis caused significant decreases in ova penetrated. A Fab preparation of these antisera exhibited similar inhibitory effects. Antisera Fab treatment of the zona-free ova prior to exposure to sperm had no effect on penetration rate.


[Characteristics of cat sensomotor cortex neuron responses to monaural and binaural stimulation].

In cats, the technique of localization parameter of dichotic sounds showed the binaural stimulation to be more effective than the monaural one. The interaural difference in intensity and time modified characteristics of neuronal responses in the sensorymotor cortex (SMC), the responses being most obvious in 67--88% of cases at the minor, approximating zero, temporal and amplitude interaural shifts. The SMC neurons altered their spontaneous activity in response to signals simulating movement of the sound source in horizontal plane. 74% of these neurons seemed to prefer the velocity up to 48 degrees/sec. These features of the SMC neuronal responses can be of major importance for formation and regulation of the cortical programs for willed motor acts of the animals in the course of acoustic orientation.


[Common pathway principle in the organization of callosal connections].

The direct cortical responses and transcallosal potentials were recorded in 170 symmetrical points of the cat motor cortex during the electrical stimulation of one hemisphere. The area of the direct responses was larger than that of transcallosal responses. The latter was concentrated in the center of the motor area, while the direct responses were located in peripheral areas. Thresholds for the direct responses were lower than those for the transcallosal responses. Variation of the strength of the stimulation led to the changes of localization of the EPs in both hemispheres. Callosotomy abolished or decreased the transcallosal potentials and increased or decreased the direct cortical potentials. This suggests the principle of the common pathway for the callosal system activity.


[Characteristics of habituation in visceral systems].

In anesthetized and immobilized cats, habituation of evoked responses to repeated stimulation of the splanchnic nerve occurred in the cerebral cortex. Upon cessation of the stimulation, the habituation waned spontaneously (spontaneous recovery). The rate of spontaneous recovery depended on number of habituating stimuli (effect of "below-zero"). Repeated series of habituation training produced a more rapid habituation of EPs (potentiation of habituation). An extrastimulus to cutaneous or splanchnic nerve leads to recovery of the evoked response (dehabituation or sensitization). Repeated application of the dehabituating stimulus produced a progressively lesser sensitization (habituation of dehabituation).


[Sympathoactivating and sympathoinhibitory afferent fibers of the vagus and splanchnic nerves].

Changes in frequency of tonic impulses in efferent sympathetic fibers of the left splanchnic nerve on stimulation (10 V, 10--20 Hz, 1 ms) of afferent fibers of the right splanchnic nerve and of both trunks of the vagus nerve at the cervical and abdominal levels have been studied in experiments on cats. It is established that there are sympatho-inhibitory and sympatho-activatory fibers whose quantitative correlation is different in various trunks. Sympatho-inhibitory influencies are most marked on stimulation of vagus nerve ventral branch and sympatho-activatory ones on stimulation of right splanchnic nerve. At the cervical level, in vagal left trunk sympatho-inhibitory fibers preponderate but in the right one another correlation occurs. In abdominal cavity the distribution of both types of fibers appears to have organ principle. Fibers innervating stomach have sympatho-inhibitory action but those innervating small intestine--sympatho-activatory. This suggests the hypothesis about the role of these fibers in digestion regulation. The central cholinolytic amysil abolishes sympatho-inhibitory effects but not sympatho-activatory ones.


[Comparative characteristics of blood and lymph clotting and fibrinolytic activity].

In dogs, the lymph coagulation occurs much more slowly than that of the blood. This is due to low concentration of the coagulation factors (I, II, V, VII, VIII, X, XI+XII) and to an increased level of heparin in the lymph. The fibrinolytic activity of lymph is somewhat higher than that of the blood which may be due to the low concentration of antiplasmines. The lymphocytes (thymocytes) prove to be able to substitute the blood platelets during coagulation of blood and lymph.


[Sequence of the spread of excitation in bird heart ventricles].

Intramural multipolar technique revealed primary sites of excitation in the subendocardium of central areas of border line of the apical and middle thirds of the bird heart septum. From the zone of primary depolarization the excitation wave spread on the subendocardium of ventricles. A number of sites of negativity were observed over the epicardium. The main mass of myocardium of free walls and the lower two thirds of the septum are activated by means of myocardium of free walls and the lower two thirds of the septum are activated by means of multifocal depolarization. The base third of the septum is the last to be activated. These features of the excitation of bird myocardium can be accounted for by the special character of distribution of the Purkinje fibers in the muscles of myocardium ventricles.


[Role of changes in the hemodynamics of the pulmonary circulation in pulmonary gas exchange].

High pressure in the lung artery was maintained with the aid of i. v. serotonin administration (0.5 mg/ml) in dogs during 10 min. The simultaneous increase of the intravascular pressure in minor circulation and of the lung volume circulation was followed by an increasing blood volume in lungs, diffusion capacity of lungs, oxygen tension in the arterial blood, and satiation of the latter with oxygen. The combined action of the above two hemodynamic factors aids to opening of latent vascular areas, to a more regular distribution of perfusion over the lung areas according to the ventilation level. This mechanism seems to be one of the first compensatory responses to disturbances in the lung gas exchange.


[Regulation of respiration in the presence of a combination of hypercapnia and an inspiratory resistance load].

The resistant load (an increased aerodynamic resistance to the gas flow) augmented both the intrathoracic pressure (ITP) and activity of the diaphragm neurons (ADN) in anesthetized cats. The increase in these parameters after alteration of CO2 tension in the alveolar gas (PACO2) by 1 mm Hg was also augmented whereas the increase of respiratory volumes (VT) was reduced. After bilateral vagotomy the inspiratory resistant load somewhat inhibited the ADN, still more so at increasing PACO2, whereas the ITP absolute values and its increase grew up. The analysis of ITP and ADN relationships suggests a significant part of intercostal and auxiliary muscles in the inspiratory increase occurring on resistant load. This becomes clearer after shunting the afferent system of lungs.


[Analysis of afferent impulsation and muscle potentials of the small intestine in response to sucrose].

Changes of potentials in the longitudinal and circular muscles of the small intestine and impulse frequency in mesenteric afferent fibers during intraluminal administration of sucrose solution were studied in cats before and after effects of adreno- and cholinolytic agents. Reserpine, dihydroergotamine, and N-cholinoblocking agents (hexonium or benzohexonium) increased the excitatory effect of sucrose whereas atropine abolished the effect. Atropine in combination with dihydroergotamine did not abolish the activating effect of sucrose. The latter seems to be unrelated to stimulation of either cholinergic or adrenergic intramural neurons. Involvement of substance P is suggested.


[Responses of individual receptor units of rat mammary gland parenchyma to tactile stimulation].

Two types of slowly adapting mechanosensitive units were revealed in the parenchyma of the rat mammary gland (SAI and SAII). At constant mechanical displacement of the parenchyma surface SAI exhibited irregular and SAII--regular firing. The diameter of the receptive fields of the SAI and SAII was 1.0--1.5 mm with the central point of this area being the most sensitive. Trapezoidal stimuli elicited both dynamic and static responses proportional to the velocity of the movement and its displacement amplitude; the threshold of the SAI and SAII increased as stimuli of increasing velocity were used.


The Sézary syndrome.

Symptoms of the Sézary syndrome are described in a case history. Sézary's syndrome is a lymphoproliferative process of the skin, which eventually develops into an erythrodermia. Strange lymphocytes with a cerebriform chromatin structure, often with T-cell characteristics, circulate in the blood stream. In few instances they are also found in lymphnodes and bone marrow. This primarily benign disease can become malignant, possibly due to an overaggressive cytostatic therapy. Besides the hitherto unknown endogenous causes, exogenous factors cannot be excluded.


Ichthyosis linearis circumflexa Comèl and Netherton's syndrome; an ultrastructural study.

An electron microscopic study of 3 cases of ichthyosis linearis circumflexa Comèl (ILC) reveals the presence of round bodies in the prickle cells: a characteristic feature of ILC. The horny and the granular layers are absent and are replaced by parakeratotic cells with condensed cytoplasm, which an amorphous material dissociates. The early formation of a vesiculopustule is detected. The importance of the dermal inflammation is considered and it is stated that ILC is not a purely epidermal disorder of keratinization.


Intranuclear particles in keratoacanthoma: possible association with malignant degeneration.

Electron microscopic investigation of 12 patients with keratoacanthoma is reported. Intranuclear virus-like particles, as described by Zelickson and Lynch, were seen in 3 cases; 2 of these subsequently underwent malignant degeneration. These findings suggest that it is particularly important to exclude the possibility of malignant change in those tumours when intranuclear particles are found.


Diffuse biphasic cutaneous amyloidosis.

2 patients with primary localized cutaneous amyloidosis were seen to have widespread macular and lichenoid types of lesions. Diffuse areas of pigmentation appeared to transform gradually over years into lichen amyloidosus as a result of chronic irritation of the skin from scratching. There was no evidence of systemic amyloidosis.


Morphaea-like cutaneous changes in a patient with systemic scleroderma.

A case of systemic scleroderma with peculiar morphaea-like lesions is reported. The patient at first started with typical acrosclerosis, some parts of which later became softened and apparently resumed the appearance of normal skin, so that morphaea-like cutaneous plaques which remained hard, were soft. These changes were surrounded by erythema resembling the 'lilac ring' as noted in characteristic active lesions of morphaea.


Low-dose chlorambucil in the treatment of generalized granuloma annulare.

5 of 6 adults with generalized granuloma annulare responded favorably to therapy with low-dose chlorambucil. 5 of the patients had lesions that were recurrent and long-standing and that resisted conventional modalities of therapy. Low-dose chlorambucil may be effective in a short-term therapy program but should be used only in patients with persistent, widespread, or unusual forms of generalized granuloma annulare. The program requires close patient and hematologic supervision.


Skin petechiae and ecchymoses (vasculitis) due to anticoagulant therapy.

The appearance of petechiae and ecchymoses following anticoagulant and fibrinolytic therapy is an unusual and rare complication. The mechanism of the vasculitis is obscure but may be related to a chemical vasotoxic reaction. During anticoagulant and fibrinolytic therapy, 6 patients developed petechiae and ecchymoses which was accompanied by pain and fever. There were no disturbances of blood clotting factors. In spite of the varied action of the different anticoagulants and fibrinolytic drugs, the clinical course and the histological examination demonstrated a similar identical vasculitis.


PUVA treatment in mycosis fungoides.

2 patients with mycosis fungoides at the plaque stage were treated with oral methoxalen and UVA irradiation. 3 months later both developed the skin tumours with highly malignant histological picture. In 1 patient at the tumour stage the skin tumours were spreading during the PUVA treatment. 3 patients at the first or eczematoid stage cleared completely or partially by PUVA treatment.


Single cilia in the articular cartilage of the cat.

Single cilia (SC) in the articular cartilage of the cat are described with regard to structure, frequency and intracellular location. Comparison is made with their occurrence in the cartilage of other species or in other tissues; special consideration is given to their developmental stages. In the 1-year-old cat examined, about 20% of the cells showed SC with a predominantly intracellular course and some of them did not reach the extracellular space at all. Cross-sections of the proximal region of SC reveal the familiary 9+0 pattern; towards the distal end, the number of doublets and the diameter of the shaft are decreasing. It is concluded that most of the SC encountered are undergoing involution.


Nonrandom correlation between selective distribution of virogenic 5-bromodeoxyuridine and DNA-binding nonhistone proteins in rat DNA.

In order to more accurately determine a mechanism for the bromodeoxyuridine (BrdU)-mediated activation of endogenous type C virus from normal rat embryo cells, nonhistone nuclear protein-DNA interactions were analyzed in vitro. Native, as well as kinetically fractioned DNA samples previously labeled with either [3H]-thymidine or [3H]-BrdU were combined with DNA-binding nonhistones and characterized according to distribution of the isotopes, and extent and localization of protein-binding sites. As before, [3H]-BrdU was relatively more concentrated in repetitive DNA as compared to [3H]-thymidine. Using a membrane filter retention assay, nearly 60% of complete, 37% of repetitive, and 12% of nonrepeated DNA-protein reconstituted complexes were retained on the filters regardless of isotopic precursor. However, a proportionately greater amount of [3H]-BrdU than [3H]-thymidine was recovered following extensive digestion of renatured complexes with DNase I, even though comparable amounts of DNA were acid-insoluble. The disproportionate binding of nonhistones to repetitive DNA, especially BrdU-substituted regions, may be related to the highly specific, well-characterized modifications in eukaryotic transcription attributed to the analog.


Inhibition by cerulenin of RNA and lipid synthesis in Limnaea (mollusc) embryos.

Cerulenin, a new antibiotic known to inhibit lipid synthesis in yeast, fungi and bacterial systems, has been tested with Limnaea embryos. In the presence of 40--50 microgram/ml cerulenin, lipid synthesis is inhibited by about 65% while RNA synthesis is inhibited by about 35% as is evident from the incorporation of 14C-acetate and 14C-uracil, respectively. In embryos treated with cerulenin, washed and returned to normal medium to which 14C-uracil was added, RNA synthesis continued normally. Such pretreatment with actinomycin, however, inhibits lipid synthesis by about 50%. Cerulenin treatment of eggs at the early morula or uncleaved stage for 30 min leads to a marked abnormality at the late veliger stage, and the embryos die before hatching. These results have been discussed in comparison with the known effects of actinomycin. As, unlike actinomycin, cerulenin has no delayed action, the present experiment offers a system for exploring the delayed effects of early transcription.


Inhibiting effect of vitamins C and B12 on the mitotic activity of ascites tumors.

The mitotic activity of the transplantable mouse tumors, Sarcoma 37, Krebs-2, and Ehrlich carcinomas, in the ascites form, were inhibited after treatment with a mixture of vitamins C and B12 with no apparent toxic side effects. These vitamins when administered alone, at the same dosage, did not seem to have any apparent effect on mitosis or the morphology of the cells studied. Microscopic examination of the stained ascites fluid taken from the mice treated with the vitamin mixture showed few tumor cells, and these in various stages of disintegration. Also, an increase in lymphocytes, monocytes and neutrophils were noticed; however, later in the experiment, no tumor cells could be found and monocytes and macrophages were abundant.


On the role of calcium in adhesion of cells to solid substrates.

Experiments are described which show that while the presence of calcium in the medium is required for the cells to maintain their adhesion, it is not necessary for the initial attachment of 3T3 cells to solid substrates. Cells are detached by treatment with urea at 4 degrees C suggesting that adhesion may involve hydrogen bonding between the cell surface and the substratum. Although most of the cell-bound calcium is removed by trypsin, the detaching effect of trypsinisation can be inhibited at low temperature suggesting that ionic calcium bridges are probably not directly involved in retaining the cells on the surface. Cells are made totally insensitive to removal by trypsin by prior washing with lanthanum. Our findings suggest that the external role of calcium in cell adhesion is exerted indirectly. We conclude that the cell presents to the exterior at least two physiochemical classes of molecule. One class composed of hydrogen bond-forming adhesive material (possible proteins) and another class of anti-adhesive molecules (possibly glycoproteins). These two components are somehow separated in the formation of adhesive 'plaques' and this process is process is apparently uninfluenced by the calcium concentration in the medium. However, the maintenance of the localised zones of adhesion is aided by factors which prevent their disruption by the intrusion into them of anti-adhesive molecules diffusing from adjacent regions of the cell membrane. These factors include cooling below the transition temperature of the membrane lipids and lateral cross-linking of non-adhesive elements by calcium. By contrast, conditions which reduce the stability of the separation of adhesive and non-adhesive surface components would be expected to diminish the overall adhesiveness of cells to the substratum.


Effects of cadmium on polyribosome sedimentation pattern in mouse liver.

Cadmium injected to mice provokes an acute disaggregation of hepatic polyribosomes. The effect appears within 15 min, it is maximal after 1 h and then gradually disappears. The reaggregation of polyribosomes takes place 6--12 h after injection. The disaggregation of polyribosomes is linear with log doses in a range of 7.25--20 mumol/kg CdCl2. Cycloheximide pretreatment prevents the disaggregation of polyribosomes in the livers of cadmium-treated mice.


In vitro expression of cytotoxicity by macrophages is independent of the actual serum concentration.

In utilizing the same effector and target cells, similar arrangement and population density of cells but varying the actual serum concentration in the range between 1 and 50% fetal calf serum, it was attempted to accurately assess the dependence of in vitro cytolysis expressed by adherent, predominantly phagocytic mononuclear cells ('macrophages') on the actual serum concentration. The results show that the extent to which cytotoxicity by activated macrophages against a given target cell type was expressed, was within a comparable range independent of the actual serum concentration. This holds true for each of the 9 target cell lines examined although with some of the cell lines there was a certain tendency towards high cytotoxicity in the range between 5 and 20% serum.


Growth of human tumor xenografts implanted under the renal capsule of normal immunocompetent mice.

The subrenal capsule technique proved effective in demonstrating that the growth of human tumors in normal, immunocompetent animals for 6 days was quantifiable in ocular micrometer units. Positive growth was demonstrable not only with human tumors that had been established in serial transplantation in athymic nude mouse hosts, but also with primary surgical explants. Growth rates of transplantation-established xenograft systems were similar whether implanted in athymic nude or in normal immunocompetent animals indicating that the 6-day time-frame successfully evades growth inhibitory effects of immunologic origin. Immunosuppression with a single dose of cyclophosphamide did not appear to affect growth rate, but permitted the tumors to grow larger extending the time to reach peak size. Significantly, xenografts of primary surgical explants showed positive growth more frequently in 6 days (82%) in the immunocompetent animal than in 11 days (30%) in the immunodeficient athymic nude mouse.


Phagocytosis and resorption during the reassembly of dissociated embryonic cells of sea urchins.

Time-lapse and electron microscopic observations were made on both epithelial and mesenchymal cells during the reassembly of embryos from dissociated cells of Strongylocentrotus purpuratus. In epithelial cells, where lysosomes are produced through the fusion of saccules formed from Golgi bodies, both phagocytosis of cell debris and resorption of differentiated cell structures were observed. In these cells, the lysosomes migrate and fuse with both autosomes and phagosomes. On the other hand, in the mesenchyme cells, where lysosomes are produced through the direct enlargement of the Golgi body's cisterna, neither phagocytosis nor resorption was observed. The migration of the lysosomes to the epithelial cell margins is the first indication of a re-establishment of cellular polarity after dissociation.


Insulin binding by a cell line (HT 29) derived from human colonic cancer.

Insulin binding was demonstrated in cultured HT 29 cells originating from a human colon carcinoma. At 37 degrees and in complete medium, the binding of [125I]insulin (1-4x10-10M) reaches a maximum in 40 min and the cell associated radioactivity remains constant for at least 4 h. No degradation of the hormone is observed under these conditions. The binding is proportional to the number of cells and its pH optimum is 7.8. In the presence of excess insulin 50% of the [125I]insulin is dissociated from the complex after 10 min. At equilibrium, insulin binding is specific: proinsulin is 25 times less potent than native insulin in competing with [125I]insulin and related polypeptide hormones are inactive. Scatchard analysis indicates two classes of binding sites (1400 sites/cell of "high affinity" e.g. 4.7 x 108 M-1, and 20 000 sites of "low affinity" e.g. 4 x 107 M-1). The binding of insulin to this non-target cell shows the same kinetic characteristics and specificity as found for insulin in its target cells, except that HT 29 cells do not degrade the hormone. The problem of the correlation between insulin binding and a biological effect in these cells remains to be elucidated.


Melatonin-and serotonin-stimulated release of vasopressin from rat neurohypophysis in vitro.

In order to study the control of vasopressin-release, the effect of a series of potential agents was studied in an in vitro perifusion system of rat neurohypophysis after in vivo treatment with nialamide, a monoamine oxidase inhibitor. In this system, metlatonin stimulated vasopressin-release in a dose-dependent manner (1 x 10-8 to 1 x 10-3 M). Serotonin (1 x 10-3 M) also led to a significant increase of vasopressin-release whereas quipazine (1 x 10-3 M), a putative serotonin agonist and monoamine oxidase inhibitor, caused a 3-fold stimulation of the release of the neurohormone. The stimulatory effects of melatonin and serotonin were prevented by omission of Ca2+ combined to an excess of Mg2+ (12mM) in the perifusion medium. 1 x 10-6 M somatostatin did not affect basal or melatonin-stimulated vasopressin-release. These results show that melatonin and serotonin can have a direct stimulatory effect on vasopressin release at the neurohypophyseal level.


Antidopaminergic activity of estrogens on prolactin release at the pituitary level in vivo.

Treatment of female or male rats with estradiol benzoate led to an almost complete reversal of the inhibitory effect of low doses of dopamine on prolactin secretion. These data indicate that estrogens which have previously been shown to exert a potent antidopaminergic activity on prolactin secretion in anterior pituitary cells in primary culture have similar effects in vivo.


Communicating vessel systems for mass cell culture of anchorage dependent cells.

Simultaneous filling of a number of vessels with identical input dosage is possible through an operation which uses the principle of communicating vessels with or without air pressure equilibration (APE). Sets of Roux flaska without APE and the NUNC Multitray Unit system with APE are presented. The latter system is used for large scale propagation of human diploid fibroblasts for production of large quantities of interferon.


Serially-propagated human diploid cells: a synopsis of the present position concerning their use or producing viral vaccines and interferon.

Diploid cells of human origin subjected to serial propagation are being used on a wide and increasing scale as substrates for producing viral vaccines for use in humans. To a lesser extent they are also being used for the production of human interferon. A brief outline of the present position on the use of these cells for producing human vaccines together with information concerning their use for the production of human interferon is presented.


Use of a new bead microcarrier for the culture of anchorage dependent cells in pseudo suspension.

The use of cross linked dextran beads having a surface charge produced using the method of Levine et al. (1976) is described. The new microcarriers require no pretreatment, and may be used at greater concentrations than those originally described by van Wezel (1967). Microcarrier cultures are initiated by inoculating cells into a stirred suspension of beads in culture medium. The cells spread over the surfaces of the beads, and follow typical growth curves. They may be harvested by conventional trypsinisation, and separated from denuded beads using a sintered glass filter. The beads may be regenerated and re-used. The beads have been found to be suitable for the growth of a variety of fibroblastoic cell types. It is suggested that the microcarriers are suited for bulk virus or metabolitic production from anchorage dependent cells.


Parameters affecting cell growth on reduced charge microcarriers.

The development of the reduced charge microcarrier and cell behavior on carriers of different degrees of DEAE substitution, and under different growth conditions are described. Parameters other than the microcarrier which limit productivity are discussed, and microcarrier growth data for certain cell types are presented.


Biological and biochemical criteria for the characterization of the transformed stage of BHK21 cells.

Some quantitative criteria for spontaneous and virus-induced transformation of BHK21 cells are described. The resistance to acid polysaccharides such as dextran sulphate in agarose medium is a characteristic shared by all tumorigenic clones. The increase in a high molecular weight integral membrane protein seems to be a biochemical feature correlated with this property.


Biochemical aspects of muscle differentiation as analyzed by in vitro cultivation techniques.

Myoblasts cultivated in vitro will undergo terminal differentiation to form muscle fibres. Teratoma derived mouse cell lines, a pluripotent primitive line, and a muscle cell line, provide a possibility for comparing RNA populations in an early precursor cell with those in committed myoblasts and differentiated myotubes. Molecular hybridization analyses led to the conclusion that new RNA sequences appear in the cytoplasm during muscle differentiation. Such muscle specific sequences are not detectable in the nuclear RNA of myoblasts or primitive cells. Studies of protein synthesis during terminal myogenesis indicate co-ordinate expression of the muscle contractile proteins. These represent distinct isozymes, distinguishable from the contractile proteins of other cell types. In the case of myosin light chains isozymic transitions between different muscle forms have been identified during early development.


Isolation of biologically active components from rabies and other envelope viruses.

Most human virus vaccines contain complete virus particles, either inactivated or attenuated. Besides components responsible for induction of neutralizing antibodies, other virus components (e.g. nucleic acids, lipids) are also administered upon vaccination. For envelope viruses the (glyco) proteins of the viral envelope are generally involved in the induction of neutralizing antibodies. Our investigations are focussed on the large scale preparation of these components from several viruses or virus vaccines, such as rabies and influenza. For virus disintegration we have tested several ionic anc nonionic detergents. Triton X-100 gave good results. Separation of solubilized components from the remainder of the virus has been carried out on a small scale by ultracentrifugation. For the purification of influenza hemagglutinin and neuraminidase we also used gelfiltration with success. The latter process can be scaled up easily. The main problem in the process of virus subunit preparation is the removal of detergent.


Training adaptations in skeletal muscle of juvenile diabetics.

Skeletal muscles from 12 male, juvenile-onset diabetics (JD) and 13 nondiabetics (ND) were studied to determine the effects of endurance training on mitochondrial enzyme activities, lipoprotein lipase (LPL) activity, and the oxidation of lipids (14C-palmityl CoA) in vitro. Ten weeks of endurance running (30 min/day, 5 days/wk) resulted in 11.0 and 12.9% gains in aerobic capacity for the JD and ND groups (P greater than 0.05), respectively. Both groups showed significant (P less than 0.05) increases in muscle LPL, carnitine palmityl transferase, succinate dehydrogenase, and hexokinase activities with training. Though the pretraining capacities for 14C-palmityl CoA oxidation were similar for both ND and JD groups, the diabetics showed a 41% greater improvement in the measurement of muscle lipid oxidation after training than did the ND group. The principal finding of this research was that skeletal muscle of juvenile diabetics who are in moderate insulin balance shows adaptations to endurance training that are similar to those of nondiabetic men.


Opposite effects of insulin and glucagon in acute hormonal control of hepatic lipogenesis.

Conditions for the isolation of rat hepatocytes that are responsive to insulin with regard to fatty acid synthesis were explored. Cells prepared according to the procedure of Ingebretsen and Wagle require the presence of fetal calf serum for insulin expression. Cells isolated by the Seglen method are the preparation of choice, since they respond to insulin in a simple, well-defined medium and, moreover, show much higher basal rates of fatty acid synthesis. In the latter cells isolated from fed male rats, the rate of fatty acid synthesis, as determined by tritium incorporation from [3H]H2O at 37 degrees C, is enhanced within 30 min after addition of insulin to the incubation medium; with glucagon, it is depressed. In the presence of insulin, the cellular content of malonyl coenzyme A is noticeably increased, whereas the concentrations of pyruvate, lactate, and citrate are not markedly affected. Glucagon, on the other hand, decreases the concentrations of all four intermediates. The activity of acetyl-CoA carboxylase is stimulated and depressed after addition of insulin and glucagon, respectively. In all conditions tested, the activity of acetyl-CoA carboxylase correlates with the rate of fatty acid synthesis, which in turn correlates with the cellular level of malonyl-CoA.


Acetylsalicyclic acid restores acute insulin response reduced by furosemide in man.

Prostaglandin E (PGE) infusion in normal man inhibits the acute insulin response to glucose. In order to determine whether endogenously released PGE might also inhibit insulin secretion, glucose-stimulated insulin responses were investigated in normal volunteers after furosemide (40 mg i.v.), a stimulator of endogenous PGE synthesis. Acute insulin response to glucose (20 g i.v.) was significantly reduced by furosemide (response before furosemide: 36 +/- 5 muU/ml; after furosemide: 26 +/- 5 muU/ml, m +/- SE, mean change 3--10 min, N = 8, P less than 0.01), whereas glucose disappearance rates were not modified after furosemide. Infusion of lysine acetylsalicylate (LAS), an inhibitor of endogenous PGE synthesis, completely reversed the inhibitory effect of furosemide on insulin secretion and also augmented acute insulin response to glucose (response before furosemide + LAS: 41 +/- 6 muU/ml; during furosemide + LAS: 50 +/- 7 muU/ml, N = 10, P less than 0.02). This effect was associated with an increase in glucose disappearance rates (P less than 0.05). These findings demonstrate that (1) furosemide inhibits glucose-induced acute insulin responses and (2) LAS completely reverses the inhibitory effect of furosemide and also accelerates glucose disposal. It is suggested that furosemide acts via the release of endogenous PGEs, which are known to inhibit insulin responses in man.


Variation in the disappearance of unlabeled insulin from plasma: studies with portal and peripheral infusions.

One hundred and thirty brief infusions of unlabeled insulin were given to 75 unanesthetized nondiabetic human beings. The patients were examined (1) by different doses of insulin (5--50 muU/kg), (2) by intraportal and peripheral infusion, (3) in the morning and in the afternoon, and (4) during normoglycemia and moderate, steady hyperglycemia. After intraportal infusion of insulin, the non-steady state plasma clearance rate (ml min(-1)kg(-1) was lower the larger the dose (decreased about 50% by a 10-fold increase in insulin dose) and lower at hyper- than at normoglycemia (decreased about 30% after small insulin doses). It was also lower the higher the relative body weight, but this correlation was weak. After peripheral insulin infusion, none of the above relationships was demonstrated. Regardless of the route of insulin infusion, plasma clearance rate did not vary with fasting plasma insulin concentration, time of day, insulin sensitivity, or glucose tolerance. We conclude that the liver modifies the distribution and removal of pancreatically released insulin in response to acute increases in insulin or glucose. Otherwise the plasma clearance rate of insulin showed a notable lack of relationship with common indexes of metabolism and with insulin action.


The association between diabetic retinopathy and skeletal muscle capillary basal lamina thickening corrected for the influence of age and duration of diabetes.

The association between an objective measure of diabetic retinopathy and skeletal muscle capillary basal lamina thickness was examined in a group of 30 male insulin-treated diabetic subjects, mean age (+/- SD) 44.6 +/- 13.2 yr, duration of diabetes 21.2 +/- 11.2 yr, % ideal body weight (% IBW) 106 +/- 11%. In addition, muscle capillary basal lamina width was measured in a group of 18 nondiabeitc men, mean age 40.7 +/- 16.3 yr and % IBW 118 +/- 23%. The muscle capillary width of the diabetic subjects was significantly greater than that of the nondiabetic group (P less than 0.01), but the values of the two overlapped considerably. In the diabetic group, there was a significant association of basal lamina width with age (P less than 0.01) but not with duration of diabetes. The association between extent of retinopathy and muscle capillary basal lamina width was not strong. The findings of the study do not therefore support the use of an estimate of muscle capillary basal lamina thickness as a single representative measure of diabetic microangiopathy.


Developmental progression in play behavior of children between nine and thirty months. I: Spontaneous play and imitation.

The developmental progression of play behavior was studied cross-sectionally in 85 children at eight age-levels between nine and 30 months. In a structured play situation, three major play patterns were distinguished: (1) exploratory play (at the end of the first year); (2) functional play (beginning of second year); and (3) representational play (after 15 months). Sequential play first occurred at 21 months. Play behavior with spatial characteristics was also observed: relational and container play between nine and 15 months, and understanding of spatial relationships, stacking and grouping between 18 and 24 months. There was a significant relationship between spontaneous and imitative play behavior; the greatest affect was displayed when the demonstrated behaviors matched the child's developmental level of play.


Intellectual development and academic achievement of children treated early for phenylketonuria.

Twenty early-treated children with classical phenylketonuria (PKU), five early-treated children with variant PKU and seven untreated children with hyperphenylalinemia were compared with non-PKU family members in terms of intellectual development, and 14 school-age PKU children were also compared for academic achievement. For the early-treated children with classical PKU, mean IQ (98) was within the normal range, but nine of these 20 children had IQ scores more than 1SD below those of family members. There was a significant negative correlation between phenylalanine concentrations at one to four years of age and later measured intelligence in these early-treated children, but this was probably a consequence of poor dietary control in the early years. The early-treated children with variant PKU and those with hyperphenylalaninemia had IQ scores consistent with those of unaffected family members, but untreated children with variant PKU had scores significantly lower than their own early-treated siblings. Achievement scores of the early-treated PKU children were consistent with their intellectual ability: they and their non-PKU siblings had similar standard scores for reading and spelling, but arithmetic scores were significantly lower for the PKU children. Early-treated children whose diet had been discontinued had achievement scores in all subjects below those predicted from their IQS.


Incidence and aetiology of infantile spasms from 1960 to 1976: a population study in Finland.

This series of cases of infantile spasms comprised 107 children born between 1960 and 1976 in the county of Uusimaa, Finland. The incidence of the syndrome was 0.38 to 0.42/1000 live births and has remained almost unchanged during those years, although there have been changes in the size of different aetiological groups. The importance of prenatal and perinatal factors, and especially of neonatal hypoglycaemia, as causal factors has decreased, whereas the numbers of cases due to infections and malformations has increased. Compared with investigations in other countries, this Finnish series differs in the incidence of familial cases and congenital cytomegalovirus infection, and especially in the higher incidence of neonatal hypoglycaemia and the lower incidence of tuberous sclerosis.


Disturbance in parent-child relationship following preterm delivery.

Increasing concern is expressed that the psychosocial development of preterm infants may be hindered by a disturbance of parental attitudes following the initial period of specialised care. Attitudes of parents of 17 preterm infants were compared with those of parents of 17 full-term infants at a single semi-structured interview six to 20 months after the birth. The groups of parents were matched for parity and did not differ in their social, ethnic and educational backgrounds. There was evidence of some disturbance in parent-child relationships in the preterm group, consisting of delays in maternal attachment to the child, negative maternal perception of the child compared with expectation of an 'average' baby, and persistent parental anxiety about leaving the child with a baby-sitter. In addition, two preterm children had been abused or neglected. Parental reaction to a preterm birth is discussed and the need for adequate support to be given to parents in the early period following the birth is stressed.


Standardisation of the Denver Developmental Screening Test for Cardiff children.

The items in the Denver Developmental Screening Test (DDST) have been restandardised on a random sample of 1547 Cardiff children aged up to six years. Some differences were found between the ages at which the Cardiff and Denver children achieved certain test items. Cardiff children were consistently slightly slower on gross motor items throughout the age-range, with the exception of three items, but were slightly more advanced on early language items below the age of 18 months and on some personal and social items. The implications of the findings for the use of the DDST in Britain are discussed.


Teaching intermittent self-catheterization to young children with myelodysplasia.

A new technique is described which enables children with neurogenic bladder secondary to myelodysplasia to learn self-catheterization prior to their school years. Inexpensive dolls, complete with male or female genitalia, and a specially designed mirror to magnify the female children's genitalia, were used with four children (three girls, one boy) with mental ages of five years. Each child successfully demonstrated self-catheterization within two days, and gradual progress toward complete independence has been promoted.


A social score for kwashiorkor: explaining the look in the child's eyes.

Following the observation that many children with kwashiorkor had a deprived look in their eyes, the guardians of 107 children with kwashiorkor and 111 controls were interviewed about their home life, in a survey made at New Mulago Hospital, Kampala, between 1969 and 1972. A child's age, sex and tribe were found to have an influence on the nutritional state, therefore 50 of each group were matched for these variables. Ten significant associations were found which distinguished kwashiorkor patients from the control group. Children with kwashiorkor were more likely to be attended by someone other than the mother; to have changed attendants when ill; to have a pregnant mother; and to have separated parents. Further to these, singletons of split partnerships were at special risk; breast-feeding had stopped; weaning was begun for bad reasons; a child was more often living away from the parents and had been sent away coincident with weaning. The fathers tended to be poor. It is suggested that these 10 factors could be used as a social scoring system in assessing the risk of incipient kwashiorkor.


Insulin secretion and pancreatic exocrine function in patients with chronic pancreatitis.

The relationship between insulin responses to oral glucose and pancreatic exocrine function were examined in 15 patients with chronic pancreatitis. Good correlations were found between the insulin responses and exocrine pancreatic function measured as the concentrations of pancreatic enzymes in duodenal juice after intravenous cholecystokinin-pancreazymin (CCK-PZ). There appears to be a roughly parallel loss of endocrine and exocrine function in the course of chronic pancreatitis.


Interrelationships of glucose and insulin uptake by muscle of normal and diabetic man. Evidence of a difference in metabolism of endogenous and exogenous insulin.

A forearm perfusion technique was used to study glucose and insulin uptake by muscle. In normal subjects at glycaemic levels above 130 mg/100 ml, glucose uptake was independent of glucose concentration;it was directly related in insulin concentration but not to insulin uptake. In non-obese maturity-onset diabetic subjects, glucose uptake was dependent on glucose concentration and insulin uptake, but not on insulin concentration. In both groups there was a strong correlation between insulin concentration and insulin uptake; diabetics had a normal insulin uptake in relation to concentration. For a given change in insulin concentration the increase in glucose uptake was as great in diabetics as in controls, but the effect of insulin was mediated through a mechanism involving its uptake. Thus in the non-obese maturity-onset diabetic, forearm muscle is not insulin resistant. The apparent uptake of insulin measured by a radioimmunoassay in relation to its arterial concentration was lower and more variable for heterologous than for endogenous insulin. With a receptor assay the venous insulin concentrations were lower than with the immunoassay and differences in uptake between endogenous and exogenous insulin disappeared. It is concluded that in muscle exogenous insulin was less severely degraded than endogenous insulin.


Fasting and two-hour post-load glucose levels for the diagnosis of diabetes. The relationship between glucose levels and complications of diabetes in the Pima Indians.

The frequency distributions of both the fasting and two-hour post-load plasma glucose levels were bimodal in the Pima Indian population aged 25 years and over. The hyperglycaemic component of this distribution represents those with diabetes mellitus, as some 30 percent of this group had evidence of the specific vascular complications of the disease, whereas these abnormalities were virtually absent in those with lower glucose levels. The bimodal characteristics of the frequency distributions were utilized to define optimal criteria to separate those with and without diabetes. The sensitivity and specificity of these criteria for fasting and two-hour glucose levels were compared and were found to be similar. The fasting glucose determination, however, was more reproducible and stable, as well as being easier to obtain, indicating that it is the better measurement for diagnostic purposes. The optimal level for diagnosis of 7.5 mmol/l (136 mg/dl) for the fasting glucose and the equivalent two-hour value of 14 mmol/l (250 mg/dl), were higher than many previously recommended diagnostic levels. Nevertheless, there was no evidence that subjects with lower levels were at appreciable risk of developing the specific complications of diabetes. Subjects with impaired glucose tolerance (IGT), but without fasting hyperglycaemia, should not be diagnosed as having diabetes mellitus.


Continuous subcutaneous insulin infusion:good blood glucose control for up to 4 days.

Six insulin-dependent diabetics were studied on their conventional insulin treatment and during continuous, dual-rate, subcutaneous insulin infusion for periods of up to 4 days. Dabetic control, as assessed by mean plasma glucose, range of plasma glucose values, M-value or range of M-values was improved significantly in 5 patients (mean +/- SD plasma glucose concentration on final infusion day 6.9 +/- 1.3 mmol/l, versus 11.3 +/- 3.2 mmol/l on conventional treatment). Once a suitable insulin dose was established blood glucose control could be maintained by continuous subcutaneous insulin infusion using the same daily infusion rate without frequent adjustment. In some case this was less than the daily dose on the conventional treatment. However, glycaemic control in one "brittle" diabetic, with unpredictable swings in blood glucose on her normal regimen, was not improved by continuous subcutaneous insulin infusion. During the period tested there was no sepsis at the cannula implantation site and patients did not find the system uncomfortable or unduly inconvenient.


Adrenal gland involvement in mice with hereditary obesity and diabetes mellitus. Morphological studies.

The contribution of the adrenal gland to the development of the spontaneous syndrome of obesity and diabetes in Yellow-KK (Y-KK) mice was studied. Six-month old Y-KK mice exhibited hyperadrenocorticism and adrenal cortex enlargement. Light microscopic morphometric studies of Y-KK adrenals revealed an expanded volume of the adrenal cortex resulting from hyperplasia of zona fasciculata and reticularis cells. Ultrastructural studies revealed fewer lipid droplets, increased numbers of mitochondria and a more extensively developed Golgi system with zona fasciculata and reticularis cells. This cytological evidence of enhanced steroid biosynthetic and secretory activity was consistent with increased levels of plasma immunoreactive corticosterone. Structural and functional abnormalities of Y-KK adrenals were preceded by the development of obesity, hyperglycaemia and hyperinsulinaemia. It is unlikely, therefore, that the adrenal plays a casual role in the syndrome's pathogenesis, although, hyperadrenocorticism may be in part responsible for an exacerbation of the observed phenomena.


Seasonal onset of insulin dependent diabetes in relation to sex and age at onset.

All newly diagnosed insulin dependent diabetics presenting consecutively to the Diabetic Department of the Royal Infirmary from the City of Edinburgh or its environs, between the years 1964 and 1977, were analysed for sex (297 males, 205 females), age at diagnosis (range 10--75 years), month of diagnosis, duration of symptoms and month of symptomatic onset. Males aged 10--19 years showed a significant seasonal variation in diagnosis (p less than 0.025) with an increase in autumn and winter months which was not seen in females aged 10--19 years nor in patients of either sex aged more than 19 years at diagnosis. The duration of diabetic symptoms increased with increasing age at diagnosis in both males and females and was consistently greater in females than in males for each age group at diagnosis. When the month of symptomatic onset was considered in relation to sex and age at diagnosis, having excluded patients with duration of symptoms of more than three months, only males aged 10--19 years at diagnosis showed a significant seasonal variation in incidence (p less than 0.005).


Blood glucose regulation using closed- and open-loop insulin delivery systems. II. Peripheral primed square wave infusions.

Glucose was infused into anaesthetized dogs before and after pancreatectomy. In the diabetics blood glucose was regulated first by closed-loop and then by open-loop insulin delivery schemes. Insulin requirements for the latter were determined by resolving the former into a sequence of 3 different infusion rates: during the baseline and recovery periods, basal insulin was delivered at 0.37 +/- 0.02 mU/kg/min, while during the 60 min glucose infusion (10 mg/kg/min) there was an 8 min infusion at 4.96 +/- 0.37 mU/kg/min and a 52 min component at 1.85 +/- 0.08 mU/kg/min. With the open-loop method under these highly standardized conditions glycaemia was similar to normal controls but IRI levels were significantly higher, 13.5 vs 8.0 microU/ml (p less than 0.05) in the baseline and recovery periods and 74 vs 25 microU/ml (p less than 0.05) during the glucose infusion. It was concluded that: constant normoglycaemia can be maintained in the basal state by a constant rate of peripheral insulin delivery but at rates resulting in peripheral hyperinsulinaemia; the glycaemic response to glucose infusion can be normalized by a two component waveform of insulin delivery; and the closed-loop method can serve as a useful guide in determining insulin requirements.


Effect of a long acting glucagon selective somatostatin analogue on plasma glucose, insulin and glucagon levels in the anaesthetized rat during arginine infusion.

The effects of somatostatin and a long acting, glucagon selective somatostatin analog (des-Ala1Gly2[His4,5-D-TrP8]-somatostatin) used studied during arginine tolerance tests in normal anaesthetized rats. Arginine infusion in control animals resulted in a rapid increase in plasma insulin and glucagon, and an increase of 15 +/- 5 mg/dl in plasma glucose. Somatostatin infusion (1 mg/kg/h) resulted in suppression of basal insulin secretion and a decrease in arginine-induced insulin and glucagon release. Glucose levels increased rapidly during the combined arginine-somatostatin infusion reaching a peak of 72 +/- 10 mg/dl above basal levels. Similar results were obtained when somatostatin was injected SC (1 mg/kg) at times 0, 15, 30, and 45 minutes (arginine infused from 30-60 minutes). A single injection (1 mg/kg) of the long-acting somatostatin analogue resulted in significant inhibition of basal insulin and glucagon release; during arginine infusion glucagon levels rose only slightly, the insulin response was, however, nearly normal, and only a small arginine-induced increase in glucose levels was observed. Carbohydrate absorption was not influenced by either somatostatin or the analogue.


The relative contributions of newly synthesized and stored messages to Hl histone synthesis in interspecies hybrid echinoid embryos.

We have measured the ratio of incorporation of 3H-lysine into the maternal and paternal forms of Hl histones synthesized by the interordinal hybrid embryo which results from the fertilization of sand dollar eggs with sea urchin sperm. This ratio has been used to calculate the relative contributions of newly transcribed and stored Hl histones mRNA to the synthesis of Hl histone at five different stages of development. These calculations are based on the assumption that histone mRNA of both parental types is transcribed with equal efficiency from the genome and that these RNAs are translated with equal efficiency in the cytoplasm of the hybrid embryos. On this basis, we have estimated that the contribution of new mRNA represents 80% of total Hl histone synthesis at the 16--32 cell stage, 54% at the hatching blastula stage, 40% at the mesenchyme blastula stage, and 100% after gastrulation. These data are discussed in the light of presently known parameters of histone and histone mRNA synthesis in echinoderm embryos.


Gel electrophoretic analysis of histones in lens epithelium, lens fiber, liver, brain, and erythrocytes of late chick embryos.

Histones from 19-day-old chick embryo lens epithelium, lens fibers, liver, brain, and erythrocytes were electrophoresed in polyacrylamide gels using buffers containing sodium dodecylsulfate, acetic acid urea, or mixtures of Triton X-100 acetic acid urea. In the last two buffer systems, histone bands were characterized by their apparent molecular weights determined by electrophoresis in the second dimension in sodium dodecylsulfate containing polyacrylamide gels. From the densitograms of the stained gels, the relative proportion of protein in different histone bands was estimated. With the exception of the erythrocyte-specific histone H5, all histones from different tissues examined at any of the gel systems migrated with the same mobilities. In lens epithelium and lens fibers, all histones were present in identical proportions. As compared to liver and brain, the total amount of histone Hl was significantly lower in lens cells and erythrocytes, possibly reflecting differences between the differentiated states. However, no tissue-specific differences were found in the relative distribution of histone Hl I and Hl II among lens epithelium, lens fiber, liver and, brain, but a threefold higher Hl I : Hl II ratio (0.5--0.7) was found in erythrocytes.


Sub-cellular distribution of the cytoplasmic myosin heavy chain mRNA during myogenesis.

In the light of earlier work [1] which demonstrated the presence of a large number of myosin heavy chain (MHC) transcripts in chick myoblasts prior to cell fusion and the burst of MHC synthesis it was of great interest to determine the subcellular localization of the still inactive transcripts. It has been determined in differentiating muscle cells in culture. Two populations of cells were examined -- monucleated myoblasts just prior to cell fusion and myotubes where at least 80% of the cells were fused. Utilizing a myosin complementary DNA (cDNA) probe [2] it is observed that just prior to cell fusion, when the "burst" of myosin synthesis has not yet occurred, the vast majority of cytoplasmic myosin mRNA transcripts are found in a stored messenger RNA protein complex with a minimal amount found in the heavy polysome fraction. In differentiated myotube cultures, when myosin synthesis is progressing at a high rate, the reverse is found, i.e, the amount of stored myosin messenger RNA (mRNA) is minimal while the largest amount of myosin mRNA transcripts are localized in the polysome fraction. The number of total cytoplasmic myosin transcripts is found to decrease after cell fusion at a time when myosin synthesis is maximal suggesting that the efficiency of translation of myosin mRNA increases during terminal differentiation.


Proteins of BrdU-dependent hamster cell lines as characterized by two-dimensional gel electrophoresis.

Cell lines which exhibit the BrdU-dependent phenotype (B4 and HAB) were studied with respect to BrdU-induced alterations in genetic expression by two-dimensional gel electrophoresis. A comparison of the proteins from the HAB cells, in which the DNA is 100% substituted by BrdU, to those of the unsubstituted parent line (3460) showed 55 protein alterations; the synthesis of 15 increased while that of the other 40 decreased. When 3460 cells were grown in BrdU such that their DNA was greater than 50% substituted, 27 protein changes could be detected; of these, the synthesis of 10 increased while that of 17 decreased. A comparison of all these changes in the various cell lines showed six which were common to the BrdU-substituted cell lines. The proteins from another Syrian hamster cell line, BHK-21 (C-13) and those of HAB cells grown in thymidine or BrdC were also examined on two-dimensional gels. Although BrdU has a dramatic effect on many cellular functions, relatively few changes in the pattern of protein synthesis could be detected in these cell lines, perhaps reflecting the specialized action of this analogue on particular cellular functions.


Telolysomes in cultured iris epithelial cells and in the TVI cell-line.

Iris epithelial cells of adult newts, which are fully differntiated melanocytes and non-dividing, become dedifferentiated and converted into lens cells when put in culture. A recent study shows that this dedifferentiation is based on an autophagic process which is associated with proliferation and mainly affects melanosomes. The present report shows that in primary culture of iris epithelial cells after the majority of melanosomes have disappeared, myelinoid bodies, which are interpreted to be telolysosomes of autophagic nature, appear in high frequencies. This suggest that in these cells autophagy persists after the loss of melanosomes. A possible connection of this type of autophagy with the differentation of lens fiber which occurs in this culture is discussed. In the TVI cell line which is believed to be derived from the same cell type, but devoid of melanosomes, similar myelinoid bodies are a characteristic cell component, suggesting that the tendency for autophagy is inherited in theis cell line.


Classes of proteins synthesized in oocytes, eggs, embryos, and differentiated tissues of Xenopus laevis.

Two-dimensional gel electrophoresis has been used to analyse protein synthesis in embryonic stages and in three differentiated tissues of Xenopus laevis. The patterns found in oocyte, unfertilized eggs, embryos shortly after fertilization and at progressively later stages of development have been characterized and compared with the patterns found in the brain, heart and liver of tadpoles. The results suggest that at least four classes of proteins can be recognized among the proteins synthesized, although other categories may exist. They also suggest that some proteins synthesized rapidly in the oocyte are likely to be synthesized in differentiated tissues as well, while proteins synthesized for the first time only after fertilization are much less likely to be synthesized in differentiated tissues.


Cell-free cytoplasmic polyadenylation of oogenic RNA.

The products of cell-free ATP incorporation mediated by cytoplasmic fractions prepared from unfertilized sea urchin eggs, anucleate egg halves, nucleate egg halves, emetine-treated fertilized eggs, and four-cell embryos have been characterized to determine to what extent the polymers synthesized are poly(A) and to assess the size distribution of the primers adenylated. As judged by alkaline lability, ribonuclease resistance, and retention on poly(U)-impregnated filters, greater than 92% of the label recovered after RNA extraction is present in poly(A). LiCl fractionation indicates that little, if any, free poly(A) is synthesized or cleaved from RNA primers during the reaction, and that 4S RNA is not an effective initiator. In excess of 85% of the poly(A) is associated with RNA having S-values greater than or equal to 18S. Sedimentation profiles of RNA adenylated in the unfertilized egg and anucleate egg half reactions are identical. Suppression of in vivo protein synthesis by emetine alters the profile of RNA subsequently adenylated in vitro. It is proposed that the apparent constraints on the utilization of cytoplasmic RNA or ribonucleoprotein primers of oogenic origin may be effected by RNA-associated proteins capable of regulating the selection and/or extent of their polyadenylation during early embryogenesis.


Mitosis in milk secreting epithelial cells of mammary gland: an ultrastructural study.

In all stages of lactation mitotic configurations were observed in mammary gland epithelial cells of rats. An electron microscopic study is presented which shows that ultrastructure of such mitotic stages is normal and that mitotic cells contain typical products of milk secreting cells such as casein micelle-containing vesicles and milk fat droplets. Such secretory products can even be observed in the immediate vicinity of the chromosomes and microtubules of the spindle apparatus. The endoplasmic reticulum of mitotic cells appeared altered in that it did not show typical cisternal stacks characteristic of interphase cells. While the numbers of such mitotic cells were very low, especially from the second week of lactation on (always less than 0.1% of the milk secreting epithelial cells encountered), the observations clearly demonstrate that differentiation for milk secretory activity and cells division are not mutually exclusive. We conclude that postpartum growth of mammary gland epithelium and replacement of epithelial cells lost during desquamation into the milk liquids can occur by division of existing differentiated milk secreting cells and does not require mitotic activity of non-lactating 'stem cells' which are not observed in lactating alveoli.


Induction of intestinal metaplasia and carcinoma in the glandular stomach of rats by N-alkyl-N'-nitro-N-nitrosoguanidines.

Induction of intestinal metaplasia and carcinoma of the stomach in Wistar rats by four homologs of N-alkyl-N'-nitro-N-nitrosoguanidine (N-alkyl-NNG) was investigated. N-Propyl-, N-butyl, N-isobutyl, and N-pentyl-NNG's were administered to rats as 0.34mM solutions in the drinking water for 12 months and the rats were killed about 6 months later. Intestinal metaplasia was found in the glandular stomach of 100%, 50%, 44%, and 17% of the rats treated with N-propyl-, N-butyl-, N-isobutyl-, and N-pentyl-NNG's, respectively. Adenocarcinoma of the glandular stomach was found in 29% of the rats treated with N-propyl-NNG, but not in those treated with other N-alkyl-NNG's. In the control group, 11% of the rats had intestinal metaplasia but none had tumors.


A new parameter for discriminating malignantly transformed cell lines from nontransformed counterparts: culture in liquid medium containing neutral protease.

Growth properties of malignantly transformed cell lines and their nontransformed counterparts were studied in the presence of a bacterial neutral protease (Dispase I). Among 10 transformed cell lines, 7 lines could proliferate in the presence of the protease in a floating state. Nontransformed cell lines and non-established cell strains failed to grow in this culture condition. The ability to grow in the presence of the protease was well correlated with their ability for anchorage-independent growth and tumorigenicity, and this provides a convenient primary test for evaluating cellular tumorigenicity.


Chronic toxicity of sodium nitrite in mice, with reference to its tumorigenicity.

Sodium nitrite has been widely used as one of the most effective food additives to tinge color on cured meat. However, it has been elucidated that this chemical is not merely a precursor of N-nitroso compounds, many of which are strongly carcinogenic, but also a mutagenic substance in biological tests. In order to ascertain the possible tumorigenicity of sodium nitrite itself, chronic toxicity of the agent in mice, by means of daily oral administration as drinking water for more than 18 months, in the concentration of 0.5 (maximum tolerated dose), 0.25, and 0.125%, was tested. As a result, development of various tumors, including thymic lymphoma, nonthymic lymphoid leukemia, pulmonary adenoma and carcinoma, and benign and malignant tumors in soft tissue, was seen in these mice. However, as to the incidence of tumors as well as the developmental time of each histologically classified tumor, no apparent difference was detected between those in the experimental groups and the control group.


Effect of BCG on cytostatic activity of peritoneal macrophages from normal and tumor-bearing rats.

In tumor-bearing rats inoculated intramuscularly with 5 x 10(4) SLC cells, the cytostatic activity of peritoneal macrophages was elevated in the early stage (7 days after transplantation) and decreased in the advanced stage (21 days after transplantation). When BCG was intraperitoneally administered into normal and tumor-bearing rats, peritoneal macrophages showed higher cytostatic activity than the untreated macrophages. This elevated cytostatic activity of the marcophages obtained from BCG-treated tumor-bearing rats was maintained even at an advanced stage.


A rapid screening for the specific DNA sequence: analysis of transforming DNA segments in adenovirus-transformed cells.

The viral DNA sequences in cells transformed by adenovirus type-12 (Ad12) DNA fragments were investigated by spot hybridization, a detection by autoradiography of nucleic acid hybrids formed between cell DNAs spotted on a membrane filter and various nick-translated Ad12 DNA fragments. In CY1 cell line, a rat cell line transformed by the EcoRI-C fragment (left hand 16%), all of the HindIII fragments included in the EcoRI-C fragment are shown to be present. In GY1 cell line, a rat cell line transformed by the HindIII-G fragment (left hand 7%), both of the BpaI-H and a part of the BpaI-J, two components consisting of the HindIII-G fragment, were found. A dominant presence of the Ad12 BpaI-H fragment (left hand end 4.5%) of the Ad12 DNA molecule, approximately 60% of the Ad12 transforming DNA sequences (Ad12 HindIII-G), was also shown in GY1. Spot hybridization used in the experiment will be of general use for detecting viral nucleic acid sequences in cells and provides a simple and useful screening method for investigating viral etiology of tumors and transformed cells.


Lymphocyte responsiveness to phytohemagglutinin and its modification by serum inhibitor in breast cancer patients.

Lymphocyte responsiveness in stimulation with phytohemagglutinin (PHA) and the inhibitory effect of autologous serum on lymphocyte responsiveness were investigated in 55 patients with breast cancer. Lymphocytes from patients, even in the late stage of the disease, as well as from normal healthy control, showed an ability to respond to the stimulation with PHA in the presence of pooled normal serum, while lymphocyte responsiveness in the presence of autologous serum was markedly reduced in stage III and IV patients. Similar result was found in the examination of lymphocyte responsiveness with relation to tumor size, where lymphocytes from patients with a large tumor of more than 5 cm in diameter were less responsive when cultured with autologous serum. From these results, it is reasonable to conclude that in patients with breast cancer, the apparent reduction of lymphocyte reactivity may be due to serum inhibitor that may adhere to the lymphocytes, rather than a primary lymphocyte impairment.


Potentiation of anticancer effect of carboquone in vivo by glucose pretreatment.

Anticancer effect of carboquone was notably potentiated in tumor-bearing mice when they were pretreated with 5% glucose solution in the system of blood flow-interrupting hyperthermic chemotherapy. It was further found by quantitative analysis that the potentiation was much greater than increase in the general toxicity of the drug by administration of glucose. Mitomycin-C, on the other hand, revealed no such marked effect though it is considerably similar to carboquone in chemical structure.


Suppression of naturally occurring anti-tissue antibodies during growth and after removal of tumours.

The sera of normal rats were found to be active in the complement fixation test against isogeneic tumour homogenates. This activity was shown to be heat labile and to sediment with IgM globulins on zone ultracentrifugation in sucrose density gradients. Sera from rats bearing progressively growing tumours and sera from rats, which had such tumours excised, were found to be less active than normal rat sera in the complement fixation test against isogeneic tumour and liver homogenates. It was considered that this suppression of naturally occurring anti-tissue antibodies may be a specific process related to the host immune response to the tumour.


Mutagenicity of quercetin and kaempferol on cultured mammalian cells.

Mutagenic activity of quercetin and kaempferol on V79 Chinese hamster cells was investigated. Quercetin was proved to have mutagenicity not only by metabolic activation system using rat liver microsomes, but also by direct application method. Kaempferol was mutagenic by metabolic activation system, but its activity by direct method seemed very low. These data suggest that the tested flavonoids seem to be metabolized for exerting their mutagenic activity and V79 cells themselves have an enzymic activation system which metabolizes at a very slow rate but in a cumulative way.


Behavior pattern of rat ascites tumor cells arrested in liver sinusoids: an electron microscopic study.

The behavior pattern of tumor cells in the development of hematogenous liver metastasis by Yoshida sarcoma and two strains of rat ascites hepatoma, AH7974F and AH13, in Donryu strain rats was investigated by electron microscopy. Tumor cells of every strain in liver sinusoids stretched their cytoplasmic protrusions toward the vascular wall and were actually in contact with the endothelial cells with their tips to form various types of junction-like structures. The most distinct one was characterized by parallel and straight arrangement of the opposing cell membranes across a gap of about 100 A. The endothelial discontinuity was easily produced by the projection of cytoplasmic processes and the compression of tumor cells. Through the endothelial defect, tumor cells of Yoshida sarcoma and AH13 migrated locomotively into the dilated spaces of Disse or directly toward the hepatic cells, while AH7974F cells were frequently covered by elongated endothelial cytoplasm. Destruction of the hepatic cells by AH7974F and AH13 cells was found to begin with the insertion of their cytoplasmic processes into the coated vesicles in the hepatic cells. From these investigations, the particular interaction between tumor and normal cells seems to play an important role in the development of liver metastasis by these tumors.


A molecular orbital study on the chemical reactivity and biological activity of polycyclic aromatic hydrocarbon diol-epoxides in connection with bay region theory.

Electronic structures of naphthalene-, anthracene-, phenanthrene-, and benzo-[a]pyrene-diol-epoxides were calculated by using CNDO/2 method together with the energy minimization method. By using these methods, the most stable geometry of molecules was automatically obtained. By comparing the total energy of phenanthrene-diol-epoxide with that of anthracene-diol-epoxide, the role of the bay region was studied in connection with the chemical reactivity. It is concluded that the bay region plays an important role when the diol-epoxide reacts with the component of the biological system via SNl mechanism. Moreover, benzo[a]pyrene-diol-epoxide was shown to be very reactive because fo the presence of the bay region as well as the large size of the molecule. When reaction proceeds via SN2 mechanism, the bay region is found to have little effect on the chemical reactivity of the molecules.


Tumor formation of rat ascites hepatoma cells in the traumatized brain.

The effect of brain trauma on the formation of metastatic growth was examined by iv injection of 6 strains of rat ascites hepatoma. Three of the tumors (AH7974F, AH66F, and AH13) were in the single-cell state and three (AH272, AH7974, and AH601) contained cell aggregates. None of the tumors showed any tumor growth in the intact brain after injection into the tail vein. However, after traumatization of the brain tissue it was possible to observe tumor formation in the lesion, mainly by injection of the single-cell type tumors. The brain tissue was most susceptive to the tumor cells 10 min and 7 days after trauma. On the other hand, after the injection into the carotid artery of traumatized animals, the tumor formation in the brain was found in both tumor groups.


Production of proliferation lesions in gastric mucosa of albino mice by oral administration of N-methyl-N'-nitro-N-nitrosoguanidine.

It was possible to produce gastric adenocarcinoma in five of 69, healthy, young albino mice of both sexes given N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) in their drinking water at a concentration of 100 microgram/ml. The tumoral lesions were most clearly defined in the group that received MNNG for the longest period of time, that is, 68 weeks. On the other hand, in the rest of the groups, which received MNNG for fewer weeks, there were lesions found in a total of nine animals which consisted in foci of typical and atypical hyperplasia, erosion of the mucosa, and, in one animal, an adenomatous polyp. Contrary to findings in humans, intestinal metaplasia preceding or accompanying the neoplasia was found to be an inconsistent alteration and a not very frequent one.


Effect of uracil on metabolism of 5-fluorouracil in vitro.

The effect of uracil on the metabolism of 5-fluorouracil (5-FU) in vitro was studied. 5-FU was mainly phosphorylated in intact Yoshida sarcoma cells, whereas it was mainly degraded in liver slices. Uracil inhibited degradation of 5-FU much more than its phosphorylation; incubation of 2,500 microM of uracil with 2.5 microM of 5-FU (molar ratio, 1,000:1) inhibited the degradation of 5-FU by 70%, but did not affect its phosphorylation. With homogenates of Yoshida sarcoma or liver uracil inhibited degradation of 5-FU greatly, phosphorylation of 5-FU by alpha-D-ribose 1-phosphate (RiblP) and ATP to some extent, and phosphorylation by 5-phospho-alpha-D-ribosyl diphosphate (PPRibP) very little. The activities of the enzymes involved in the metabolism of 5-FU in various tissues were also determined. Degradation of 5-FU was much faster in liver than in other tissues and was very slow in tumor tissue. Phosphorylation of 5-FU with RiblP and ATP was rapid in Yoshida sarcoma and bone marrow. Phosphoribosyltransferase activity was high in Yoshida sarcoma and thymus, but low in bone marrow.


Transplantable ileal adenocarcinomas of ACI rats.

Transplantation of 9 kinds of bracken-induced ileal adenocarcinomas subcutaneously in ACI rats established two transplantable strains of adenocarcinoma (73-357, 77-238). Ascitic conversion succeeded in the strain 73-357. Biological and morphological, particularly electron microscopic, characteristics of these strains are reported.


Tumor development in lung of ddY mice following transplacental exposure to 1-ethyl-1-nitrosourea.

Transplacental induction of lung tumor by 1-ethyl-1-nitrosourea (ENU) was studied in pregnant ddY mice which were given a single intraperitoneal injection of 58.5 mg/kg of ENU in water between day 13 and 19 of gestation. Within 4 approximately 6 weeks after birth, pulmonary tumor nodules were found in all offsprings exposed to ENU, and they were histopathologically adenoma. Number of tumor nodules could be counted under the stereomicroscope from approximately day 40 after birth. The size of tumor increased with the lapse of time but the number of tumor nodules did not increase markedly. Weekly injections of urethan or ENU into mice pretreated with ENU in their fetal age enhanced the number of pulmonary adenoma. The development of other tumor was not seen except a few cases of lymphoma. Tumor development in the lung by injection of ENU in ddY mice during gestation is reproducible, relatively simple, and rapid. Therefore, it is considered that this may be a useful method for screening of antitumor agent.


Nucleation time: a key factor in the pathogenesis of cholesterol gallstone disease.

In earlier studies, we concluded that biliary cholesterol supersaturation may be a necessary but not sufficient cause for gallstone formation. In the present studies, we calculated cholesterol saturation indices (CSI) for 120 bile specimens of cholesterol gallstone patients and controls, using the solubility boundaries of Carey and Small (J Clin Invest 61:998-1026, 1978) for artificial biles. The mean CSI + SD was 1.42 +/- 0.68 for controls and 1.80 +/- 1.02 for cholesterol gallstone patients. Of the control bile samples, 68% were supersaturated. Since the two groups could not be sharply distinguished by saturation index, we studied another property of bile samples: nucleation time for cholesterol crystal formation. The mean nucleation time for 16 control bile samples was 15 days, and a strong correlation (r = 0.84) was found between CSI and nucleation time. Twelve bile samples of cholesterol gallstone patients had a mean nucleation time of 3 days and no correlation between CSI and nucleation time. A discriminant analysis of the data, which took into account both the CSI and the nucleation time for each sample, allowed a sharp distinction between bile samples of cholesterol gallstone patients and controls. This distinction could not have been drawn from studies with artificial biles, since these differ only in their CSI.


Successful medical treatment of pyogenic liver abscess.

Between 1976 and 1978, 6 patients with proven or probable pyogenic liver abscess were successfully treated at Los Angeles County-University of Southern California Medical Center without surgical drainage. Although surgical therapy is necessary in patients with coexisting intraabdominal pathology, antibiotic therapy without surgical intervention is an option to be considered in patients with pyogenic liver abscess and no evidence of other intraabdominal disease.


Control of upper gastrointestinal hemorrhage by endoscopic spraying of clotting factors.

A new technique for the endoscopic control of upper gastrointestinal hemorrhage has been developed. It consists of a spraying device which delivers atomized clotting factors (thrombin and fibrinogen) directly to a focal bleeding site. In vivo studies in a standardized dog model of acute gastric ulcer bleeding, demonstrated a significant reduction in bleeding time by spraying thrombin and cryoprecipitate (mean +/- SE bleeding time = 223 +/- 14 sec) as compared to a saline spray (mean 705 +/- 60 sec) or to the untreated controls (mean 498 +/- 43 sec), P less than 0.001. Similar results were obtained in the heparinized dog model. The mean bleeding time was 226 +/- 11 sec in the sprayed animals, while the control ulcers bled continuously. The four lumen spray tube fits the biopsy lumen of commercial gastroscopes. The feasibility of the technique for treatment of oozing venous bleeding sites in the upper gastrointestinal tract has been successfully tried in a clinical setting. The authors feel that the technique in its present form is ready for clinical testing in a controlled manner.


Cholesterol gallstone formation and prevention by chenodeoxycholic and ursodeoxycholic acids. A new hamster model.

Prior animal models of cholesterol gallstone formation have been criticized for their dissimilarity to the conditions of humans with gallstones. We present a new hamster model of cholesterol cholelithiasis that more closely approximates the human situation. Sixty female Golden Syrian hamsters (average weight 83.2 +/- 3.4 g) were allocated to six groups of 10 animals each. Groups were fed standard diet (containing 0.8 gm cholesterol/g of food) or increased cholesterol diet (containing 2.4 mg cholesterol/g of food), with or without ethinyl estradiol, 15 micrograms/kg/d. Two groups receiving both increased cholesterol and ethinyl estradiol also received either chenodeoxycholic acid or ursodeoxycholic acid, 20 mg/kg/d. The animsl were sacrificed at 12 wk. Cholesterol gallstones (78.3 +/- 5.0% cholesterol by weight) formed in 30% of the animals fed ethinyl estradiol, 50% of those fed increased cholesterol, and 90% of those fed the combination of both. Bile was saturated in all three groups, with the saturation index of the combination group (2.08 +/- 0.17) being the highest. In both groups receiving bile acid therapy, no gallstones were found, and the bile remained unsaturated. For the bile acid-fed groups, both hepatic HMG-CoAR and hepatic cholesterol 7 alpha-hydroxylase activities were reduced (P less than 0.01) when compared to the group fed standard diet and to the grou fed the combination. Thus, a new animal model of cholesterol gallstone formation has been developed in which chenodeoxycholic acid and ursodeoxycholic acid therapy prevented gallstone formation through mechanisms similar to those reported in cholesterol gallstone patients.


Progressive systemic sclerosis of the gastrointestinal tract and hereditary hollow visceral myopathy: two distinguishable disorders of intestinal smooth muscle.

The purpose of this study was to determine whether progressive systemic sclerosis (PSS) of the gastrointestinal tract and hereditary hollow visceral myopathy are two distinguishable disorders of intestinal smooth muscle. We coded and blindly reviewed 50 specimens of tissue from the small intestine of 15 normal controls, 4 patients with visceral myopathy, 5 patients with PSS and intestinal pseudoobstruction, and 5 patients with PSS but no pseudoobstruction. We determined that there is a fundamental difference between the pathology of these two disorders. In visceral myopathy, the smooth muscle is characterized by vacuolar degeneration and fibrosis. In PSS, smooth muscle fibrosis is present but vacuolar degeneration is absent. Although smooth muscle cells are decreased in number in both disorders, those that are present in PSS are morphologically normal by light microscopy, whereas most of those present in visceral myopathy are degenerated. Visceral myopathy and gastrointestinal PSS are two distinct disorders of smooth muscle which are easily distinguished by conventional light microscopy. Their dissimilar appearances and the familial nature of visceral myopathy suggest that they result from quite different causes.


Effect of vagotomy in Zollinger-Ellison syndrome.

We evaluated the effect of vagotomy on gastric acid secretion and the clinical course in 3 patients with Zollinger-Ellison syndrome. Basal acid hypersecretion was reduced by 49, 86, and 96%, and peak acid output in response to pentagastrin was reduced by 36, 39, and 71% in the 3 patients. In one patient, 300 mg cimetidine reduced basal acid secretion from 65 to 20 meq/hr before vagotomy; whereas after vagotomy basal acid secretion was reduced from 36 to 0.6 meq/hr by the same dose of cimetidine. One patient has required no antisecretory therapy for 14 yr, whereas 2 patients have also been treated with cimetidine with excellent results. We conclude that vagotomy facilitates control of acid secretion in Zollinger-Ellison syndrome, and we recommend vagotomy and cimetidine rather than total gastrectomy or cimetidine alone for the management of these patients. This combined surgical and medical approach should also allow discovery and removal of isolated tumors in about 10% of patients.


Release of somatostatin-like immunoreactivity from the lower gut.

The present study describes the effect of the instillation of a nutrient-containing hyperosmolar volume load into the lower ileum upon plasma levels of somatostatin-like immunoreactivity. The baseline levels of SLI in the mesenteric vein were significantly higher than those in the inferior vena cava and rose significantly in both veins in response to the nutrient load. SLI levels in the venous effluent of the pancreas and stomach did not rise. Mesenteric vein SLI was similar in molecular size to the SLI in the gastric and pancreatic veins and to synthetic somatostatin. The results suggest that the lower gut may contribute to the basal circulating SLI levels and that it is released from the lower gut in response to a nutrient-containing hyperosmolar volume load.