Microbial skin flora of selected cancer patients and hospital personnel.

The bacterial flora of the skin from five anatomical sites on 10 leukemia patients, 10 patients with malignant melanoma, and a control group of 10 medical personnel was examined quantitatively and qualitatively. This was done to determine whether malignant disease results in changes in skin flora and to establish carrier rates of gram-negative bacteria on the skin of personnel in hospital environments. Gram-negative bacteria were isolated more frequently (74 isolates from 100 cultures) from the skin of leukemia patients than from either patients with malignant melanoma (8 isolates from 100 cultures) or the medical personnel (9 isolates from 100 cultures). Klebsiella pneumoniae and Pseudomonas aeruginosa were isolated exclusively from leukemia patients. Relative proportions of gram-negative bacteria in total populations were determined. The axilla was the only site with a uniformly high proportion of gram-negative bacteria. From all other sites cultured, gram-negative populations were low (1 to 5 bacteria/cm2 of skin), although a high proportion of gram-negative populations occurred randomly throughout all subject groups. It was concluded that leukemia patients tend to carry gram-negative bacteria on the skin. The factors permitting colonization of skin by gram-negative bacteria are discussed.


Evaluation of a rubella hemagglutination inhibition test system.

Two systems for measurement of rubella hemagglutination inhibition antibodies were compared. One had chick erythrocytes as indicator and the other, which is available in kit form ("Rubindex," Ortho-Diagnostics), uses human group O erythrocytes. Correlation to within one dilution of each other was noted in 92% of the tests. The Rubindex system has the advantage of longer shelf life of reagents and also does not require one of the absorption steps necessary for the other method, thus saving technologist's time.


Effect of short-term, low-level nitrogen dioxide exposure on bronchial sensitivity of asthmatic patients.

Our purpose was to determine whether exposure to a realistic concentration of nitrogen dioxide (NO2) could increase the bronchial sensitivity of asthmatic patients to bronchoconstrictor agents. We established dose-response curves for changes in specific airway resistance (SRaw) in response to aerosolized carbachol in 20 asthmatics after each had spent 1 h in an exposure chamber breathing on one occasion unpolluted air and on a separate occasion 0.1 ppm NO2: sequence of exposures to unpolluted air and to low levels of NO2 were randomized in a single-blind fashion. NO2 induced a slight but significant increase in initial SRaw and enhanced the bronchoconstrictor effect of carbachol in 13 subjects: curves were shifted to the left and the mean dose of carbachol producing a twofold increase in initial SRaw was decreased from 0.66 mg to 0.36 mg (P less than 0.001). In contrast, NO2 neither modified the initial SRaw nor the bronchoconstrictor effect of carbachol in seven subjects. In 4 out of the 20 subjects, exposure to a higher concentration of NO2 (0.2 ppm) yielded variable results. Potentiation of the carbachol bronchoconstrictor response by NO2 could not be related to any physical or clinical characteristics of the subjects tested. Although the mechanisms underlying the NO2 effect remain controversial, the present results demonstrate that very low levels of NO2 can adversely affect some asthmatics.


Kinetic analysis of biliary lipid excretion in man and dog.

To understand better the mechanisms involved in biliary lipid excretion and to evaluate their role in cholesterol gallstone formation, the rates of biliary excretion of bile salts, cholesterol, and phospholipids were measured in two species, man and dog. Seven cholecystectomized patients with balloon-occludable reinfusion T-tubes were studied during intact and interrupted enterohepatic circulation and four cholecystectomized dogs were studied during interrupted enterohepatic circulation. In man and dog both cholesterol and phospholipid outputs were hyperbolically related to bile salt output by the equation y = x/(a + bx). The output curves intersected the origin and showed an initial rapid rise, followed by a slower increase to a maximum, suggesting a rate-limited mechanism. The shape of the curves permitted calculation of the theoretical maximal outputs and the rates of rise to those outputs. Comparison of these values showed that in both man and dog phospholipid output was greater than cholesterol output and that cholesterol and phospholipid were excreted at different rates. These studies (a) indicate that cholesterol, phospholipids, and bile salts are not excreted in a fixed relationship and (b) demonstrate the usefulness of the derived theoretical maximal lipid output, and the rate of rise of lipid excretion to a maximum, in evaluating the kinetics of biliary lipid excretion.


Effect of intravenous furosemide on the renal excretion of digoxin.

The effect of an 80-mg intravenous dose of furosemide on the urinary excretion of digoxin was determined in three adult men with normal renal function, each of whom was taking 0.25 mg digoxin daily on a chronic basis. On two separate days, serum samples were taken and urine was collected every 2 hours over an 8-hour period for determination of digoxin, creatinine, calcium, and sodium concentrations. On the first day of study, a saline bolus was given intravenously, and on the second day, furosemide was given. In all subjects, urinary digoxin excretion increased after furosemide in direct proportion to the increase in urine volume. No consistent correlation was seen between digoxin excretion and creatinine, calcium, or sodium output. No significant changes in serum digoxin were found in this active study. These results are consistent with the hypothesis that increasing glomerular filtration rate or total urine volume increases the renal excretion of digoxin and may result in increased total urinary output of this glycoside.


Therapeutic doses of cyproheptadine do not inhibit monoamine oxidase in man.

Cypro, a serotonin and histamine antagonist, has been shown to be a moderately potent reversible inhibitor of tissue monoamine oxidase (MAO) obtained from hamsters and rabbits. In the present study, Cypro inhibits MAO obtained from human platelets with the same potency as harmine (Ki = 5 x 10(-5) M). However, when ten normal volunteers received conventional therapeutic doses of Cypro (16 mg/day) for two days, there was no alteration in their urinary excretion of tryptamine, 5-hydroxyindoleacetic acid, 3-methoxy-4-hydroxymandelic acid, epinephrine, or norepinephrine. We conclude that, when used in conventional clinical doses, Cypro does not inhibit MAO in man.


Serum creatine phosphokinase concentrations after intramuscular chlordiazepoxide and its solvent.

Serum creatine phosphokinase (CPK) concentrations were determined in healthy volunteers for the first 48 hours after intramuscular injection of 50 mg chlordiazepoxide hydrochloride or of its solvent alone. Intramuscular injection of both the drug solution and its solvent was painful and caused CPK elevations. The CPK rise due to the drug solution was 33 per cent higher than that due to the solvent alone, but the difference was not significant. The pH of the solvent preparation is low and it contains high concentrations of propylene glycol. The pain and muscle damage due to injection of the solvent could be due to its acidity and its high osmolarity. Problems associated with intramuscular injections of water-insoluble drugs are not resolved by the use of such solvent preparations.


Norepinephrine infusion in normal subjects and patients with essential or renal hypertension: effect on blood pressure, heart rate, and plasma catecholamine concentrations.

Infusion of NE in seven normal subjects and 13 patients with essential or renal hypertension caused a pronounced initial rise of systolic pressure in only seven hypertensives and one normotensive. This hyperresponsiveness was not a constant finding in essential or renal hypertensives but usually occurred in patients with highest preinfusion pressures. In some of the latter, following the pronounced rise in pressure when NE infusion was started (0.05 mug/kg/min), pressure did not increase further (probably due to reflexly reduced cardiac output) despite progressively increasing the infusion rate to 0.1 and 0.2 mug/kg/min. Hyperresponsiveness could not be attributed to increased NE concentrations at receptor sites since it was not significantly correlated with elevations of NE plasma concentrations; in some essential hypertensives, mild pressure increases occurred despite marked elevations of arterial plasma NE. Since hyperresponsiveness occurred in some patients with essential and some with renal hypertension, it could not be used to differentiate these two groups of hypertensives. The mechanism for hyperresponsiveness remains unclear but may be better explained by vascular structure alterations than by hyperreactive vascular smooth muscle per se; however, a combination of these factors could participate. During NE infusion, reflex bradycardia was associated with elevated pressure and was slightly more pronounced in normotensives; this was probably related to diminished baroreflex sensitivity in essential hypertensives and due to "resetting" of their baroreceptors. During high rates of NE infusion (0.2 mu/kg/min), higher arterial plasma NE concentrations in essential hypertensives than in normotensives could result from reduction in blood flow to organs important in inactivating circulating NE; however, a defective inactivating mechanism for NE in some essential hypertensives cannot be totally excluded.


Hypochondriasis in the elderly: a reaction to social stress.

Hypochondriasis in the elderly is often a preventable or reversible syndrome. It can become chronic if the patient finds no relief from social stress or becomes dependent upon medical services as a source of support. When this is recognized, psychotherapeutic intervention is necessary. Although no socioeconomic group is exempt, hypochondriasis in the elderly occurs more often among the lower social classes. Its higher prevalence in this group is attributable to the frequency and severity of social stress and the loss of alternative social opportunities. If psychotherapeutic intervention is necessary, the elderly hypochondriac patient should be helped to recognize social stress as a major source of the problem and to develop a realistic method of coping with it. Apparently the precipitating factors are often in the socioeconomic sphere; hence, social planners should be aware of this fact if the demands on the health care system are to be reduced.


Goals of the cytology automation program of the National Cancer Institute.

The immediate goal of the Cytology Automation Program of the National Cancer Institute is the development of an automated system which will screen appropriate specimens from asymptomatic women for squamous cell carcinoma of the cervix and its precursor lesions, dysplasia and carcinoma in situ. This system should make one of three decisions on each specimen: normal, abnormal or inadequate with an acceptably low false negative rate. It is expected that the next step in evaluation of patients whose specimens have been identified as abnormal by the automated system would be a manual evaluation of a traditional cytologic specimen.


Spectral studies on 33258 Hoechst and related bisbenzimidazole dyes useful for fluorescent detection of deoxyribonucleic acid synthesis.

Absorption, fluroescence and circular dichroism measrements on 33258 Hoechst-deoxyribonucleic acid (DNA) complexes are consistent with the existence of two types of dye-binding interactions. One type, which persists at elevated solution ionic strength, is highly specific for adenine-thymine-rich DNA. Dye bound under this condition exhibits efficient fluorescence and strong optical activity. A less specific, largely electrostatic interaction is associated with less intense fluorescence and weaker optical activity. The fluorescence of 33258 Hoechst and several other bisbenzimidazole dyes is less when bound to poly(deoxyadenylate-5-bromodeoxyuridylate) than when bound to poly(deoxyadenlyate-deoxythymidylate). Quenching of 33258 Hoechst fluorescence can also be used to detect biosynthetic incorporation of 5-bromodeoxyuridine into the DNA of living cells. This property of 33258 Hoechst should allow fluorescence-activated cell and chromosome sorting according to the extent of DNA synthesis, providing a bridge between biochemical and cytologic analyses of processes related to DNA replication.


Gynecologic specimen analysis by multiangle light scattering in a flow system.

A flow-system instrument is described in which the laser light scattered by a mammalian cell is sampled simultaneously at up to 32 angles between 0 degrees and 21 degrees from the laser beam axis as the cell passes through the beam. The scatter pattern for each cell is stored by a computer for later analysis. Various data-processing techniques are discussed. Results of preliminary application of the instrument to the analysis of normal and abnormal gynecologic specimens are presented.


Immunologic reactivity to soluble autochthonous tumor extracts in dogs with spontaneous malignancies.

The presence of tumor-associated antigens (TAA) in dogs with spontaneous lymphoma (L) or solid tumors (ST) was investigated by testing soluble tumor cell extracts for their ability to stimulate autochthonous in vitro lymphocyte blastogenesis and in vivo intradermal delayed hypersensitivity responses. Tumor extracts were prepared from tumor biopsies and control extracts were prepared from buffycoat cells of tumor dogs by hypertonic KCl solubilization. Blastogenic responses to intact, cryopreserved, autochthonous tumor cells were also determined. Fifteen of 26 (58%) L dogs and 23 of 32 (72%) ST dogs had positive blastogenic responses to at least one concentration of autochthonous tumor extracts. Blastogenic responses to tumor extract and to cryopreserved whole tumor cells correlated well when the tests were performed simultaneously. No consistent effect of autochthonous serum on the lymphocyte response to autochthonous tumor extract was observed. Two of 11 L dogs showed a positive skin test to autochthonous tumor extract and none reacted to control extract. Three of 21 ST dogs showed a positive skin test and two had a borderline response, while two ST dogs had a positive response to control extract. However, five of seven L and nine of 18 ST dogs were anergic as defined by failure to respond to intradermal PPD challenge after sensitization with Bacille Calmette-Guerin. In conclusion, lymphocytes from most dogs with spontaneous tumors underwent blastogenesis in vitro in response to autochthonous soluble tumor extracts, and these responses when tested simultaneously correlated well with responses to cryopreserved intact tumor cells. These results are compatible with the presence of TAA in extracts from cells of spontaneous canine tumors. Delayed skin test reactivity, however, was an uncommon and unpredictable finding, and does not appear to be useful in detecting TAA in the dog.


The induction of hapten-specific immunologic tolerance and immunity in B lymphocytes. I. The effect of delayed immunization on the adoptive response to TNP-LPS.

An improved adoptive transfer procedure has been developed for the thymus-independent conjugate TNP-LPS. This technique involving delayed immunization results in increased sensivity and makes possible studies of various cell types, the response of which is normally very low or which are difficult to manipulate experimentally in situ. The standard adoptive immune response of adult spleen cells to TNP-LPS was low in comparison to the primary AFC response of intact mice. Adult bone marrow cells gave a 30-fold lower and neonatal spleen cells an 8-fold lower adoptive response than an equivalent number of adult spleen cells. If the administration of antigen was delayed past the normal time of immunization (1 hr after cell transfer), the resulting AFC response of adult spleen, adult bone marrow, and neonatal cells was enhanced. The peak response occurred with a delay interval of 4 days, and the magnitude decreased with greater delay. Part of the reason for the decline was an acceleration in the attainment of a peak response in mice given antigen past the 4-day optimum. The enhancement of the adult splenic response was not restricted to the antigen TNP-LPS, and was a transferrable phenomenon which did not require antigen to persist past the 4-day optimum delay period. The ineffectiveness of host pre-irradiation indicated that host recovery alone was not involved. Although the results are more compatible with B cell proliferation causing the enhancement, a combination of effects may be involved.


Accumulation of glyceryl ether lipids in Wolman's disease.

We have shown that ether-linked glycerolipids accumulated in the adrenal, liver, and spleen of a male Chinese infant with Wolman's disease; the increases were mainly in the alkyl and alk-1-enyl glycerolipids that did not contain phosphorus. Alkyldiacylglycerol accounted for a portion of the rise in the neutral alkylglycerols. The spleen also contained increased amounts of ether-linked phosphoglycerides of the alkyl and alk-1-enyl types. Organs from a Niemann-Pick patient were also included in this study; they did not show comparable rises in the content of ether-linked glycerolipids, suggesting the possibility that storage of these compounds may be characteristic of Wolman's disease, or a variant form thereof.


Biochemical changes in the corpus luteum of the cyclic hamster.

The corpora lutea (CL) of the hamster secrete progesterone on days 1 and 2 of the oestrous cycle before marked regression at pro-oestrus (day 4) virtually eliminates them from the ovary. Luteal protein and RNA reached their maximum values on day 2 and declined up to day 4. DNA was constant throughout the cycle substantiating the importance of cellular hypertrophy rather than hyperplasia in the growth of the CL. Concentrations of cholesterol and its esters were unchanged between days 1 and 3 but almost doubled in concentration on day 4. Glucose-6-phosphate dehydrogenase activity was maximal on days 2 and 4, apparently associated on these days with steroidogenesis and lipid metabolism, respectively. A significant increase in acid phosphatase activity occurred on day 3 reaching maximal values on day 4, probably accounting for the rapid regression of the CL. The maximal concentration of 3beta-hydroxysteroid dehydrogenase in CL was on day 1, with a gradual decline over the next 3 days.


Developmental, diurnal and oestrous cycle-dependent changes in the activity of liver enzymes.

The metabolism of [4-14C]4-androstene-3, 17-dione was studied in the 105000 g microsomal and supernatant fractions of liver from developing rats of both sexes. The following enzyme activities were measured: 5beta-reductase (supernatant fraction) and 5alpha-reductase, 17alpha- and 17beta-hydroxysteroid reductases, 6beta-, 7alpha- and 16alpha-hydroxylases (microsomal fraction). The activities of the 3alpha- and 3beta-hydroxysteroid reductases were estimated by calculating the ratios of 3alpha-: 5alpha- and 3beta-: 5alpha-reduced metabolites formed, respectively. Most enzyme activities present at birth (i.e. 5beta-reductase, 5alpha-reductase, 17beta-hydroxysteroid reductase, 6beta- and 7alpha-hydroxylase) increased until 20 days of age in both male and female rats. Between 20 and 30 days of age a number of masculine metabolic characteristics appeared in both sexes, i.e. the 16alpha-hydroxylase and the 17alpha-hydroxysteroid reductase were induced, the 5beta-reductase activity rapidly increased and the 5alpha-reductase activity slightly decreased. During a third period beginning 30 days after birth the adult male enzyme activity pattern was completed by the induction of 3beta-hydroxysteroid reductase and a further increase in the activity of 16alpha-hydroxylase. After 30 days of age a feminine type of liver metabolism also rapidly developed in female rats; the 16alpha-hydroxylase and the 17alpha-hydroxysteroid reductase activities disappeared, the 6beta-hydroxylase and the 5beta-reductase activities decreased and the 5alpha-reductase activity increased six times. The developmental patterns of enzyme activities in the rat liver are consistent with a first developmental phase (0-30 days of age) independent of hypophysial control and probably determined primarily by the genome of the liver cell and a second phase (from 30 days onwards) with increasing sexual differentiation under hypophysial control. This control is mediated by some kind of feminizing factor in female rats and possibly by some kind of androgen-elicited secretion of masculinizing factor(s) in male rats. The metabolism of [4-14C]4-androstene-3, 17-dione was also studied during different times of the day and during different phases of the oestrous cycle. The 16alpha-hydroxylase activity showed a diurnal variation with higher values at noon than at midnight. The 5beta-reductase activity reached a maximal activity during metoestrus.


Effects of sodium pentobarbitone on serum gonadotrophin levels and on the incorporation of 35S from methionine into protein in the brain and anterior pituitary during the oestrous cycle of the rat.

Ovulation was delayed for 24 h after the administration of sodium pentobarbitone (Nembutal, 35 mg/kg body weight) at 14.00 h, before the critical period on the afternoon of pro-oestrus. The expected preovulatory surge of serum LH at 18.00 h of pro-oestrus was also delayed until 21.00 h on the following day; however, increased levels (less than 12 ng/ml) were observed in 14 out of 23 animals (killed by decapitation) at 21.00 h on the day of Nembutal administration. The serum FSH rise observed on the morning of expected oestrus was extended after Nembutal treatment, and a further rise was noted 24 h later. Peak levels of incorporation of 35S from methionine into protein of the median eminence area (ME) and of the anterior pituitary (AP) which normally occur about the time of the preovulatory LH surge, were also delayed until 21.00 h on the day following Nembutal administration. Neither ovulation nor the preovulatory gonadotropin rises with their accompanying changes in incorporation in the ME and the AP, were altered by Nembutal administered after the pro-oestrous critical period. Thus Nembutal, while blocking ovulation, inhibits the circadian rhythm of incorporation of 35S from methionine in the brain as well as the peaks of incorporation in the median eminence and the anterior pituitary which accompany the normal preovulatory surges of gonadotrophin.


Studies on androgen transport into canine prostate in vitro.

Slices from canine prostate were superfused with [17alpha-3H] testosterone and 5alpha-dihydro[1,2-3H]testosterone at 'physiological' concentrations; to some wuperfusions, either [3H]cyproterone or [3H]cyproterone acetate was also added. The following parameters were measured: 'uptake' of anti-androgen by the tissue, rate of entry of testosterone and 5alpha-dihydrotestosterone into the tissue, rate of efflux from the tissue of 5alpha-dihydrotestosterone and concentration of 'diffusible' 5alpha-dihydrotestosterone in the slices. The adsorption of steroids on to the surface of the slices and the bulk flow of tritiated water into the slices were also investigated. It was concluded that these two factors did not interfere with the measurement of entry and efflux of the androgens. Cyproterone and cyproterone acetate were not metabolized, but were concentrated in the slices to many times the level in the medium. With rate of supply of anti-androgens up to 43 pmol/min, their uptake increased together with the rate of entry of the two androgens and the rate of efflux of 5alpha-dihydrotestosterone. At a higher rate of supply of anti-androgens, their uptake and the rates of entry and efflux of the androgens decreased sharply. In most cases, the inward movement of 5alpha-dihydrotestosterone into the slices took place against a negative concentration gradient, while that of testosterone always occurred down a positive concentration gradient. These results confirmed the hypothesis already put forward that 'carriers' involved in the transport of androgens are present in the membrane of prostatic cells (Giorgi, Moses, Grant, Scott & Sinclair, 1974). Non-specific and specific intracellular binding, exhibiting greater affinity for 5alpha-dihydrotestosterone than for testosterone, was demonstrated in canine prostate by means of 'washing-out' experiments. On the basis of its affinity for androgens and the presence of inhibition by low concentrations of anti-androgen, intracellular binding seemed to be due to components distinct from the postulated membrane 'carriers'.


Parameters of electrical stimulation of the medial preoptic area for release of gonadotrophins in male rats.

The release of LH and FSH after the application of an electrical stimulus to the anterior diencephalon of male rats has been studied. The stimulus was applied through either platinum or steel electrodes implanted stereotaxically in animals anaesthetized with urethane. The efficacy of various parameters of stimulation by means of a current consisting of balanced biphasic square waves, was tested by systematically changing the frequency, amplitude and duration of the pulses. The effect of direct current (d.c.) stimulation on hormone release was also examined. The concentrations of the hormones in blood withdrawn from the femoral vein before and at frequent intervals up to 80 min after application of the stimulus were determined by radioimmunoassay. The optimal parameters for the release of LH by square wave stimulation of the medial preoptic area were: frequency, 60 Hz; pulse amplitude, 0-50 mA; pulse duration, 1-00 ms. This stimulus was more effective when applied through steel than through platinum electrodes. Direct current stimulation (15 muA for 10 s) through steel electrodes was most effective of all. When applied through platinum electrodes to the medial preoptic and anterior hypothalamic areas, the optimal square wave stimulus produced significant increases in the concentration of LH after 5 and 10 min respectively. The concentration of plasma FSH in these animals also increased, but the increments were much less than the increments in LH. The magnitude of the respective increases of the gonadotrophins after stimulation of the two brain areas did not differ significantly. Measurement of the milk ejection response to stimulation of the hypothalamo-hypophysial tract in the lactating rat indicated that the spread of the square wave stimulus was no more than 1-5 mm from the electrode tip. The significance of these findings is discussed with respect to the importance of the medial preoptic area in the male rat, the neurones which may be involved in the regulation of gonadotrophin secretion, and the parameters of stimulation used in studying the hypothalamo-hypophysial system.


Leteinizing hormone and luteinizing hormone releasing hormone-like immunoreactivity in the jugular venous blood of sheep at various stages of the oestrous cycle.

Luteinizing hormone and LH-RH-like immunoreactivity were measured in the jugular venous plasma of Clun Forest ewes at various stages of the oestrous cycle. Blood samples were collected through jugular venous cannulae every 2 h for at least 20 days from three ewes during the breeding season. The ewes were checked twice daily for oestrus using a vasectomized ram. Plasma LH peaks of apparent height 112-192 ng NIH-LH-S17 equivalents/ml were detected at oestrus with basal levels of 2-15 ng/ml during most of the remainder of the 17-day oestrous cycle. Peaks of LH-RH-like immunoreactivity occurred at various times of the cycle. The apparent maximal level of these peaks was 220 pg/ml compared with basal levels of less than 10 pg/ml. Further ewes (two for each group) were sampled at 4 min intervals for 12 h, (1) from onset of oestrus, (2) 36-48 h after onset of oestrus or (3) on day 10 of the oestrous cycle. In the ewes sampled at oestrus, peaks of LH-RH-like immunoreactivity were detected before, during and after the preovulatory LH peak. Those detected after the LH peak were unassociated with any further increases in the plasma LH level. In the ewes sampled 36-48 h after onset of oestrus and on day 10 of the cycle, several peaks of LH-RH-like immunoreactivity unassociated with any increases in the LH level were detected. These peaks, and those detected at oestrus, had durations of only one or two samples, and in some cases reached levels of several ng/ml compared with basal levels of less than 10 pg/ml. The significance of these results is discussed.


Effects of sodium pentobarbitone administration on gonadotrophin release, first ovulation and ovarian morphology in pubertal rats.

Pubertal female rats received sodium pentobarbitone (PB; 45 mg/kg body wt) at various hours on the day of first pro-oestrus. Maximal blockade of ovulation, in about 60% of the rats, occurred after PB treatment at 12.00, 13.00 and 14.00 h. The number of small antral follicles (100-499 X 10(5) mum3) was reduced 1 day after PB treatment in both blocked and ovulating rats. In the ovaries of non-ovulating rats signs of stimulation by LH such as dispersion of cumulus cells, oocyte maturation and early luteinization were sometimes present; in ovulating rats cystic corpora lutea with entrapped ova were found in addition to normal corpora lutea. Gonadotrophin measurements after PB treatment (14.00 h) in pubertal and adult rats showed (at 17.00 h) reduced levels of both LH and FSH, these levels being lower in the adults. Gonadotrophin levels of blocked and ovulating pubertal rats overlapped. In PB-treated, pubertal rats in which ovulation was postponed by 1 day, vaginal oestrus was prolonged by 1 day and the subsequent dioestrus by 2 days. The pubertal rat is thus less sensitive to PB treatment than the adult. PB treatment of the younger animal influences not only the ovulatory process but also follicular growth and, presumably, the length of the approaching cycle.


Effect of intra-uterine beads alone, or combined with oestrogen treatment, on utero-ovarian venous plasma prostaglandin F2alpha and progesterone levels in the guinea-pig.

The presence of beads in the guinea-pig uterus caused premature regression of the corpora lutea, and a corresponding decrease in utero-ovarian venous plasma levels of progesterone, by days 9-10 of the oestrous cycle. If the beads were placed in one horn only, premature luteal regression occurred only in the adjacent ovary. Induced luteal regression was preceded by an increased release of prostaglandin F2alpha (PGF2alpha) from the uterine horns which contained the beads. Oestradiol benzoate treatment of guinea-pigs carrying beads in one uterine horn overcame the unilateral effect and resulted in premature luteal regression in both ovaries. This was also associated with the early release of PGF2alpha from both uterine horns, though a difference in the timing of the release was apparent between the bead-containing (experimental) and control horns.


Serum gonadotrophins and follicular development in immature rats after early androgen administration.

Follicular development and serum gonadotrophin levels were studied in female rats after neonatal androgen administration. After injection of 1250 mug testosterone propionate (TP) on day 5 after birth the composition of the follicular population was altered: at nearly all ages the number of pre-antral follicles (follicular volume 2-20 X 10(5) mum2) was lower than in oil-treated rats, in some cases the number of small antral follicles (21-249 X 10(5) mum3) was also lower. Furthermore levels of serum follicle-stimulating hormone and luteinizing hormone were decreased from day 7 to day 20 suggesting that the high gonadotrophin levels before day 20 are of importance for normal follicular development. In contrast, final follicular maturation in TP-treated rats was enhanced; at day 35 more large antral follicles (follicular volume greater than or equal to 500 X 10(5) mum3) were present in TP-treated rats than in oil-treated rats. The presence of more large antral follicles was accompanied by higher plasma oestradiol concentrations, higher uterine weights and advanced vaginal opening. These results demonstrate an inhibition of normal follicular growth and an acceleration of ovarian maturation after neonatal androgen administration.


Sex reversal in a freshwater sponge.

Spongilla lacustris exhibits a type of alternative hermaphroditism, new to the phylum, in which a sponge may be exclusively male or female during the period of sexual reproduction one year and the opposite sex the next year. This form of sexuality may facilitate larva production and thus dispersal following colonization of a new habitat. Gametogenesis occurs shortly after gemmule hatching in both males and females but slightly later in males.


Carbon dioxide fixation in Helminthosporium cynodontis.

A mycelial suspension of Helminthosporium cynodontis (ATCC24938), grown on glucose-peptone-yeast extract broth and exposed to NaH14CO3 for 5 h, fixed significant quantities of 14C into the following fractions (%): small molecular weight components, 7-4; lipid and lipoproteins, 3-9; nucleic acids, 59; the residual protein and cell wall fragments, 29-2. The labelled protein components were (%): aspartate, 39; glutamate, 18; cystine, 15; threonine, 9. Radioactive nucleic acid components were (%): adenine, 18; guanine, 18; cytidylate, 34; uridylate, 30. When the mycelium was grown in Czapek-Dox glucose medium and incubated in this medium plus NaH14CO3, the nucleic acid fraction contained 29-9% and the residual protein 49-5% of the cellular radioactivity. The removal of CO2 from the atmosphere did not reduce growth. Pyruvate carboxylase (PC) and phosphoenolypyruvate carboxykinase (PEPCK) activities were demonstrated in extracts of H. cynodontis. Synthesis of PEPCK was stimulated under conditions promoting gluconeogenesis and was reduced under conditions promoting glycolysis, while PC synthesis was similar under both conditions.


Role of choline in the nutrition of the rumen protozoon Entodinium caudatum.

A requirement of choline for growth of Entodinium caudatum in a simplified culture medium has been demonstrated. Ethanolamine, N-methylethanolamine, or N-dimethylethanolamine were ineffective as substitutes. In the rumen, the normal environment of this organism, levels of free choline were virtually zero even after ingestion of pasture containing phosphatidylcholine which was rapidly catabolized. Free [Me-14C]choline is very rapidly cleared from rumen fluid, a little being incorporated into the phosphatidylcholine of protozoa, but the clearance also occurs in animals with defaunated rumens. It is suggested that E. caudatum obtains choline for growth mainly from plant membrane material which it has ingested, rather than from the free base in the rumen liquor.


A note on the relationship between the comfortable interpersonal distance scale and the sociometric status of emotionally disturbed children.

The purpose of this study was to investigate the relationship between two measures of social attraction or status: namely, the recently developed Comfortable Interpersonal Distance (CID) Scale and a traditional measure of sociometric status. Both measures were recorded for a sample of emotionally disturbed, preadolescent boys who were in residential treatment. The results indicated that the two measures were very highly correlated. This finding means that the more a boy was liked by his cottage peers, the closer these peers indicated that they would like to sit next to the boy in a room. In other words, physical interpersonal distance seems to be nonverbal way of communicating liking for a person.


Heart rate and avoidance conditioned activity in rats.

In accordance with the determination of base-line heart rate and running in an activity wheel, 14 Long-Evans male rats were given ten 30 minute sessions of Immobile-Avoid conditioning followed by 30 minute sessions of Active-Avoid conditioning. Control Ss were yoked to the experimental Ss. Experimental Ss made significantly fewer responses in the immobile-avoid period and significantly more responses in the active-avoid period than did the yoked control Ss. Heart rate was correlated with skeletal activity both for experimental and control Ss. For experimental Ss, low heart rate was concomitant with a low amount of activity in the immobile-avoid peroid and high heart rate was concomitant with greater skeletal activity in the active-avoid period. These results support the position that heart rate is secondary to responses of the somatic-motor system and that heart rate during immobile-avoid or active avoid conditioning is more closely related to the level of somatic-motor activity than to the emotional stress which purportedly motivates instrumental avoidance behavior.


The California Psychological Inventory and the Semantic Differential dimensions.

The feasibility of describing first order factors and scales of the California Psychological Inventory (CPI) by using the factors of the semantic differential and rated adjustment and introversion-extraversion was investigated in samples of 150 male and 80 female college students. Results indicated that both the semantic differential and the ratings significantly overlapped with the CPI, but that the degree of the relationship was not large enough to support the contention that the semantic differential factors are sufficient to describe completely what is being measured by the CPI. The relationships did prove useful, however, in further clarifying the nature of the CPI.


Cerebral circulation and EEG alpha frequency in relation to daily fluctuations in psychotic behavior.

Aspects of the cerebral circulation were monitored daily by cerebral electrical impedance plethysmography in one chronic schizophrenic girl for 9 months and seven other chronic schizophrenic patients for a period of 8 months. An additional manic-depressive patient was monitored while in a remission phase over 2 months. Simultaneous electroencephalogram (EEG) recordings were made on the schizophrenic patients. Daily Malamud-Sands (MS) psychiatric rating scales were completed by trained nursing staff on all patients. Correlation coefficients were obtained between this behavioral variable and the EEG-measured alpha frequency. Correlations were also run between the dimensions of the MS rating scale and three indices of the cerebral circulation and between two indices of cervical blood flow. Results indicated significant correlations between the "stuporous" scores of the MS scale and the cerebrovascular measurements in one catatonic patient. EEG alpha frequency correlated significantly with "excitement" scores in the same patient. The manic-depressive patient monitored in her remission phase showed no correlations with MS scores but relatively unchanging normal cerebral hemodynamics. The remaining seven chronic schizophrenic patients were followed daily for 8 months during a drug regimen in which phenelzine and L-tryptophan were administered. Significant relationships were found between the daily fluctuations of behavior and cerebral circulatory variables in a majority of the patients, and the extent of these correlations appeared to be increased by the addition of the metabolically active agents. Only a minority of correlations between MS scores and EEG dominant frequency were found in this group of patients. It is concluded that the MS scale affords a useful ethological monitor of psychotic behavior and that cerebral electrical impedance plethysmography and EEG alpha frequency correlate significantly with the fluctuations it records.


Female responsiveness to erotic films and the "ideal" erotic film from a feminine perspective.

Subjects were 20 volunteers randomly selected from all female undergraduate students attending a local university, who were 21 years of age or older, and living off campus. The stimuli consisted of seven films, of approximately 5 minutes duration each, which depicted various types of human sexual behavior. Themes included "romantic" heterosexual behavior, primarily genital heterosexual behavior, a mild, and an explicit form of group-sex behavior, a mild and explicit form of heterosexual sadomasochistic behavior,  and male homosexuality. Subjects rated the degree of sexual arousal experienced for each film on a 50-point Likert scale. The order of presentation of the films for each subject was randomized, and all subjects participated individually with only a female experimenter present. Following the rating period, subjects completed the Sex Knowledge and Attitude Test (9). Finally, a structured interview was conducted to probe specific likes and dislikes with respect to the content of the films viewed. Results indicated that rank order ratings of the films, from most to least sexually stimulating, was: heterosexual, romantic; group-sex mild (two males and one female); heterosexual-genital; group-sex explicit (three males and three females); sadomasochism (mild); sadomasochism-explicit (forcing and brutal); and male homosexuality. One conclusion that the data led to was that females preferred and were significantly more sexually stimulated by films in which a male related to a female (even if the male was treating the female cruelly) than they were by homosexual stimuli involving two males together. Correlational data between the rating responses and the Sex Knowledge and Attitude Test yielded much information including significant negative correlations between ratings of both group-sex films and rejection of sexual myths. Also, significant positive correlations were found between rated degree of sexual stimulation for the heterosexual and the group-sex films and the consideration of pre- and extramarital relations as acceptable or even desirable.


A case of camptocormia occurring in psychotherapy.

Although camptocormia has been known to be a functional symptom since its first description in 1837, there has been little opportunity to study the psychodynamics of this symptoms. The author describes the development of this symptom in a patient who had been in psychotherapy for 3 1/2 years and explores the meaning of the symptom for this patient. The author makes suggestions as to proper therapeutic management of patients with this symptom.


Motor-unit activity responsible for 8- to 12-Hz component of human physiological finger tremor.

Tremor of the extended third digit and bipolar surface and needle electromyograms of the extensor digitorum were recorded from six healthy volunteers for the purpose of elucidating the motor-unit activity responsible for the 8- to 12-Hz component of physiological finger tremor. Tremor was measured with a force transducer during steady voluntary contractions of approximately 200-250 g. The surface EMGs were full-wave rectified and low-pass filtered (-3 dB at 21 Hz), producing the envelope of the surface EMG (the demodulated EMG). Spectral analyses of simultaneous tremor and demodulated EMG records were performed. In four of six subjects, a pronounced 8- to 12-Hz amplitude modulation in the surface EMG was present, and coherency analysis demonstrated that this modulation was strongly correlated with the well-known 8- to 12-Hz tremor. In two subjects this amplitude modulation and tremor were barely detectable, despite the sensitive recording and analysis techniques used in this study. Spectral analysis was performed on 43 motor-unit spike trains. Twenty-two spike trains, having mean firing frequencies in the range of 10-22 spikes/s, produced statistically significant spectral peaks at 8-12 Hz, in addition to the expected spectral peaks at the mean firing frequencies. Of the 22 8- to 12-Hz-producing motor units, 12 had mean firing frequencies in the range of 17-22 spikes/s and exhibited the greatest 8- to 12-Hz activities of all motor units recorded. These motor units displayed transient sequences of double discharges in which interspike intervals (ISIS) of approximately 8-30 ms alternated with ISIS of 60-90 ms, thus producing an 8- to 12-Hz spectral peak. Adjacent ISIS of these motor units were correlated in the range of -0.5 to -0.9. Coherency analyses demonstrated that the 8- to 12-Hz activities of these motor units were correlated with the 8- to 12-Hz finger tremor and surface EMG modulation. The remaining 10 8- to 12-Hz-producing motor units had mean firing frequencies in the range of 10-17 spike/s. Although these motor units did not display the intense double-discharge firing pattern of the more rapidly firing motor units, a tendency toward action potential grouping was present and resulted in 8- to 12-Hz spectral activities which were correlated with the tremor and surface EMG modulation. .. ..


Jugular venography for evaluation of abnormalities of the skull base.

Jugular venography done to evaluate abnormalities at the base of the skull demonstrated three distinctly different patterns depending on whether there is occlusion, invasion, or growth with in the internal jugular vein. Improper technique results in a lack of intracranial dural sinus filling which may masquerade as venous occlusion. This problem is avoided by adequate neck compression along with proper volume and rate of delivery of contrast. Radiographically, an abnormal jugular vein at the base of the skull will show a concave defect in true occlusion, constriction or invasion of the vein by tumor, or tumor growth within the vein.


Pituitary failure secondary to head trauma. Case report.

A patient presented with hypopituitarism, 2 years after severe head trauma. Deficits of growth-hormone, follicle-stimulating hormone, luteinizing hormone, and borderline thyroid-stimulating hormone (TSH) were demonstrated. Normal TSH-releasing hormone (TRH) response and elevated prolactin indicated viable anterior pituitary tissue with inadequate hypothalamic control. Precautions are suggested for recognition and treatment of this syndrome.


Influence of pyridoxine supplementation on vitamin B-6 levels in milk of rats deficient in the vitamin.

Levels of vitamin B-6 in milk from pyridoxine deficient dams were used as an indicator of the ability of pyridoxine to protect offspring against the effects of the deficiency. Sprague Dawley rats were fed a basal diet containing 30.0 (control) or 1.2 (deficient) mg pyridoxine-HC1/kg diet from weaning throughout growth, gestation and until 5 days postpartum. At this time, deficient dams were supplemented by a single intraperitoneal injection of 600 mug pyridoxine-HC1, or by adding 30 or 60 mg pyridoxine-HC1/kg to the diet. The vitamin B-6 content in milk form the group supplemented by injection exceeded the control level of 38.8 mug/100 ml milk 30 minutes after the injection, and reached a peak level of 110.7 mug/100 ml at 4 hours with a subsequent decline to 27mug/100 ml at 20 hours. In rats supplemented orally with 30 or 60 mg pyridoxine-HC1/kg diet, the vitamin B-6 level in the milk reached the control value in 24 and 6 hours, respectively. At 120 hours, orally supplemented dams had significantly higher levels of vitamin B-6 in the milk than control animals. Vitamin supplementation of dams by a single injection of pyridoxine-HC1 was sufficient to overcome the pyridoxine deficiency syndrome in the pups, but was not adequate for optimum growth.


The influence of dietary inositol on glyceride composition and synthesis in livers of rats fed different fats.

The effect of inositol supplementation on the composition and biosynthesis of glycerides in the livers of rats fed diets containing fats with differing fatty acid composition was investigated. The dietary fats employed in these studies included corn oil, Tower rapeseed oil (RSO), partially hydrogenated soybean oil (SBO), and tallow. No significant influence of inositol on hepatic triglyceride levels was found in animals fed corn oil and SBO whereas inositol deficiency caused a two-and four-fold elevation in triglyceride concentrations in the RSO and tallow groups, respectively. The level of total fatty acids in phospholipid (mg/g liver) was slightly decreased in all fat groups and included a notable decrease in the concentration of monoenoic (18:1) and dienoic (18:2) acids in the RSO and tallow groups. Dietary inositol had a minor effect on the weight % of individual fatty acids in hepatic triglycerides. Inositol-deficient animals showed an increased rate of glyceride synthesis from glycerol-3-phosphate (glycerol-3-P) regardless of the type of fat that was fed. The results indicate that triglyceride accumulation in liver under conditions of inositol deficiency is not only produced with highly saturated fats since the most unsaturated of all the fats tested, Tower RSO, also gave the syndrome. The results further suggested that the accumulation of hepatic lipid due to the feeding of inositol-deficient diets likely arises from their effect on specific metabolic site(s) other than the conversion of glycerol-3-P into lipid.


Effects of different levels of dietary iron on pregnancy superimposed upon growth in the rat.

The effects of feeding four levels of dietary iron, 10, 50, 250 and 1.250 mg/kg were studied during pregnancy in growing and adult rats. Hematological measurements, plasma iron and total iron binding capacity, and iron content in liver, spleen and tibia were compared relative to pregnancy, diet and growth. Iron content in fetuses and fetal livers were compared relative to diet and growth of the dams. All parameters were lowest in rats fed the 10 mg Fe/kg diet. The highest level of iron fed, 1,250 mg/kg, resulted in increased iron content in liver, spleen and tibia of all treatment groups but did not alter hematological values or fetal iron content. Pregnant rats fed any of the four levels of iron had significantly lower Hb, Ht, total and storage iron concentration and ferritin and hemosiderin iron in liver than nonpregnant rats fed the same levels. The level of dietary iron needed by growing pregnant rats for maximal iron content in fetuses and fetal livers was between 50 and 250 mg Fe/kg which was fivefold that needed to obtain maximal hemoglobin concentration in dams. However, adult pregnant rats only needed between 10 to 50 mg Fe/kg for both maximal iron stores in fetal tissues and maximal hemoglobin concentration.


Scanning electron-microscopic study of the surface characteristics of neoplastic endothelial cells of blood vessels.

The morphological surface characteristics of neoplastic endothelial cells of blood vessels induced by oral administration of 1,2-dimethylhydrazine dihydrochloride in mice were studied by scanning electron microscopy. The interconnecting vascular channels were lined by malignant endothelial cells which formed bridges, cord-like structures and had extended cytoplasmic processes which made the network more intricate. The endothelial cells were often in close approximation to each other and exhibited irregular growth patterns and orientation. The cell surface displayed a rugged, terrace-like microcontour of elevations, depressions, blebs and nodular structures. These findings correlate with the earlier morphological observation obtained by transmission electron microscopy.


Renal tubular necrosis due to shock: light and electron-microscope observations.

The renal biopsy findings in a 76 yr-old woman suffering grom anuria due to acute tubular necrosis are described. The glomeruli were normal on light- and electron microscopy. Immunofluorescent studies failed to reveal any fibrin or immunoglobulins in the glomerular capillaries. Extensive focal areas of necrosis were seen in the tubular epithelium often exposing the lumen of the tubule directly to the tubular basement-membrane. In some areas necrotic cells lay adjacent to normal or near normal cells. The proximity of the necrotic tubular epithelium to the oedematous interstitial tissue and the peritubular capillaries, together with the finding of normal glomeruli is compatible with the theory of back diffusion as a mechanism for the oliguria.


Melanotic tumours (Blue Naevi) of spinal nerve roots.

Four cases interpreted as intraspinal blue naevi are reported. The patients were adults females with an age range between 22 and 60 yr. In three there was a single tumour arising from the cervical posterior nerve roots and in the fourth there were multiple tumours arising from the posterior nerve roots of the spinal cord and occurring within the cerebello--pontine angle. The histological appearances of the tumours were similar in every way to those of dermal blue naevi. One was of the more common spindle-celled type and three of the cellular variant. The tumours contained melanin-pigment, and spindle cells with dendritic bipolar processes of the type described in dermal blue naevi. One was of the more common spindle-celled type and three of the cellular variant. The tumours contained melanin pigment, and spindle cells with dendritic bipolar processes of the type described in dermal blue naevi. Definite evidence of malignant tranformation was found in two cases and in a third, the appearances were suggestive for early malignant change. Therefore, unlike their dermal equivalents, intraspinal blue naevi appear to have a greater propensity for malignant transformation. In each case a careful clinical examination failed to reveal any evidence of a primary malignant melanoma. In the one case who died and on whom necropsy was performed, the failure to identify a primary cutaneous, mucosal or ocular melanoma substantiated our contention that these tumours were primary.


Cerebrospinal fluid endotoxin concentrations in gram-negative bacterial meningitis.

The limulus lysate assay was utilized to detect and quantitate endotoxin in cerebrospinal fluid from 232 patients with suspected meningitis. The assay was positive in initial specimens of CSF from all 86 patients with gram-negative bacterial meningitis and was uniformly negative in the remaining 146 patients with a variety of infectious and noninfectious processes. Endotoxin concentrations in initial specimens of CSF from patients with gram-negative meningitis ranged from 4 to 2,000 ng/ml. No correlation between initial CSF levels of endotoxin and initial clinical or laboratory variables of infection was noted. With antibiotic therapy, CSF concentrations of endotoxin fall rapidly to undetectable levels after five days.


Neonatal septic arthritis.

Neonatal septic arthritis of the knee was encountered in nine infants during a two-year period, representing an incidence of 16.5 per 1,000 admissions to a neonatal intensive care unit. The etiologic agents included Staphylococcus aureus (3), Klebsiella (1), Streptococcus, group B (2), and Candida albicans (3). Two infants with systemic fungal infection died. Arthritis was the presenting feature of neonatal septicemia in three of six infants with bacterial infection and was accompanied by osteomyelitis in two. In eight of the infants, the same organism was cultured from the skin of the umbilicus as was cultured from the joint. Umbilical catheters had been placed in all of these infants. Full recovery of joint function has been noted in the seven surviving infants.


Feed the nursing mother, thereby the infant.

A successful attempt to improve nutritional status of infants by supplementing their mothers' diet is described. The habitual diet of a chronically malnourished woman whose milk production was insufficient to feed her newborn infant was supplemented with extra calories. An increase in milk production and in the infant's weight was rapidly obtained.


Creativity and body image boundaries.

Descriptions of the creative individual or the individual who can regress in the service of the ego seem to parallel Fisher and Cleveland's (1968) description of the high barrier individual: well-integrated; self-sufficient, but often outgoing and communicative; and able to experience a full range of sensation and emotions without neurotic anxiety. The relationships between Barrier and five measures associated with creativity were investigated, using 100 paid male college student volunteers. The results support the notion that creativity is not a functional unity. Barrier was found to be related to aristic, expressive creativity which seems  to be related to interest in human interactions, but unrelated to creativity associated with scientific endeavors and unrelated to creative receptivity (i.e., purest adaptive regression).


Oxygen solubilization by lung surfactant.

To illustrate the concept of solubilization as a possible mode of gas transport in biological systems, dog lung surfactants in varying concentrations were tested for their ability to solubilize oxygen. The degree of gas solubilization was determined by GC, using a modified tonometer as an absorption chamber. The concentration of surfactant was found to be an essential factor for gas solubilization. Surfactant concentration above the CMC yielded anomalously high gas absorption. Solubilization of the gas is thought to occur by a partitioning effect into the interior of surfactant micelles.


Oral ingestion of narcotic analgesics by rats.

Rats drank solutions of narcotic analgesics as the only drinking fluid available in their cages. Relative to their daily water intake before the drug solutions were introduced, the rats drank more etonitazene solution, less methadone solution, and about the same volume of morphine, meperidine and levorphanol solution as water, although some rats would not drink the higher concentrations of morphine (1.0 mg/ml), levorphanol (1.0 mg/ml) and methadone (1.0 mg/ml). When naloxone (1.0 mg/kg) was administered after 12 days of drinking the drug solutions, the severity of the abstinence syndromes based on scoring of symptoms and on weight loss was morphine greater than or equal to etonitazine greater than levorphanol greater than meperidine greater than methadone. The animals showing the most severe syndrome after naloxone usually consumed more drug solution than usual during the next 24 hours, so that body weights were normal at the end of the 24-hour period. Nonprecipitated abstinence was also studied in these rats by replacing the drug solutions with water 3 to 5 days after the naloxone injection. Rats that had been drinking morphine, etonitazene and levorphanol lost weight for 2 or 3 days and then began to regain their lost weight. In contrast, rats which had been switched from methadone and meperidine solutions to water gained weight rapidly. Morphine and etonitazene drinkers which had been switched to water drank a lesser volume of water than they had been drinking of drug solution whereas some ex-methadone drinkers drank more water.


In vivo actions of clozapine and haloperidol on the turnover rate of acetylcholine in rat striatum.

We have measured acetylcholine (ACh) content and turnover rate (TRACh) in striatum and cortex of rats receiving haloperidol and clozapine i.p. Both clozapine (30 mumol/kg) and haloperidol (10 mumol/kg) reverse the decrease in striatal TRACh elicited by apomorphine (11 mumol/kg) while each antipsychotic affects the steady state and the TRACh in striatum differently. Haloperidol fails to change striatal ACh content but increases the TRACh; chozapine (15 and 30 mumol/kg) neither decreases the content of ACh nor changes the TRACh in striatum. Moreover, 60 or 90 mumol/kg of clozapine causes a 40% decrease in ACh content without affecting the TRACh. Clozapine, but not haloperidol, antagonizes the increase in ACh content and the decrease in TRACh elicited by arecoline (64 mumol/kg) and oxotremorine (9 mumol/kg) in striatum. Clozapine resembles trihexylphenidyl (14 mumol/kg) and benztropine (12 mumol/kg) because it decreases the ACh content of striatum without changing the TRACh. Moreover, clozapine and benztropine reverse the increase in striatal TRACh elicited by haloperidol. The increase in striatal TRACh elicited by haloperidol could be of value to explain the extrapyramidal action of this drug. The anticholinergic action of clozapine could explain the absence of extrapyramidal side effects observed with this drug.


Membrane currents of the tunicate egg under the voltage-clamp condition.

1. Ionic currents of the egg membrane of a certain tunicate. Halocynthia roretzi Drashe, were studied by the voltage-clamp technique. 2. The membrane depolarization beyond -55mV in standard artificial sea water induced mainly transient inward current and slight outward currents, when the holding potential was kept at -99 mV. 3. The transient inward current was composed of two components; the major one showed a faster time course, a more negative critical level of about -55 mV, and a reversal potential around +60 mV and the minor one showed a slower time course, a less negative critical level o -10 mV, and no definite reversal potential. 4. The major component became maximum at about -25 mV with the peak time of 6-9 msec at 15 degrees C, and the maximum currents ranged from 0-5 to 1-5 X 10(-5) A/cm2. 5. The major component of the inward current was abolished by the replacement of Na with choline or Tris or Cs ions, while it was almost unaltered by the replacement with Li. The minor component was independent of Na concentration in the external solution. 6. The major component showed the activation and inactivation identical with those of Na current of other excitable membranes. A conditioning depolarization over -90 mV inactivated the Na current and the half inactivation of the major inward current was obtained by a conditioning pulse to -56 mV, when the pulse duration was 400 msec and the temperature was at 15 degrees C. 7. The time course of the Na current was formulated with m and h parameters in the following equations: (see article). 8. The kinetic parameters taum and tauh of egg Na current were calculated and compared with those of the squid axon. The potential dependence of taum and tauh was almost identical with that of the axon, but the absolute values of both taum and tauh were ten- to twentyfold larger than those of the axon in any range of the membrane potential. 9. The temperature depdence of the kinetic parameters taum, tauh and of the chord conductance gNa was studied. The Q10's for taum and tauh were both 4-0, while the Q10 for gNa was 2-0 in the temperature range from 5 to 20 degrees C. 10. The outward and inward rectifying conductances of egg membrane were remarkably activated at the potential level above +100 mV and below -70 mV respectively in standard artificial sea water. Both increased currents were subsequently subject to inactivation. 11. It was suggested that Na, Ca, K inward rectifying and K outward rectifying conductances all exist separately in the egg cell membrane and the Na current was essentially identical with that through the Na channel in other excitable membranes.


A diurnal rhythm in the absorption of glucose and water by isolated rat small intestine.

1. Glucose and water absorption by isolated small intestine from rats which have had unrestricted access to food is 50-60% higher at night than during the daytime. 2. When the feeding time is restricted to 06.00-09.00 hr G.M.T. glucose and water absorption rates in the period from 3 to 7 hr after withdrawal of food are almost as high as the rates observed at night-time in the animals with unrestricted feeding. 3. These changes in absorption rates appear to be associated with feeding time and not with the pattern of illumination.


[Sacro-iliac osteochondritis (author's transl)].

Sacro-iliac osteochondritis is difficult to recognize, it is rather frequently a case of diagnosis by elimination, which can be only confirmed by the course. Its prognosis is excellent. The predominance of lesions on the iliac edge reflects the affinity of this epiphysitis for the accessory iliac ossification point.


Cyclic activity of the pituitary prolactin cells and plasma prolactin levels in the oestrous cycle of the ewe.

The relative proportions of prolactin cells, somatotrophs and gonadotrophs were determined in the adenohypophyses of 28 Merino ewes on selected days during the oestrous cycle. Plasma prolactin levels were measured in 16 of the animals at 3-hr intervals during the 24 hr before autopsy. From Days 1 to 5 of the cycle (Day 0=day of ovulation), plasma prolactin levels were 20-40 ng/ml and the prolactin cells were filled with granules. Plasma levels increased to 168 ng/ml between Days 6 and 9, and marked degranulation of the prolactin cells occurred. The greatest degranulation was found on Days 16 and 0, and was associated with a prolactin surge to a concentration of 610 ng/ml. The rise in plasma prolactin and intensive degranulation of prolactin cells at the time of ovulation and formation of the CL suggest that prolactin is important at this stage of the oestrous cycle.


Capillary blood flow to ovarian follicles, stroma and corpora lutea of anaesthetized sheep.

The radioactive microsphere technique was used to determine the rate of blood flow through vessels of up to 15 mum diameter in the ovaries of 23 anaesthetized sheep in the 72 hr preceding ovulation. The validity of the microsphere technique was established in two preliminary studies. On Days 14, 15 and 16 of the cycle the rate of blood flow (ml.min-1 .100 g-1 tissue) was 1122, 708 and 116 to the CL; 157, 258 and 140 to the stroma; and 627, 742 and 1096 to the follicles, respectively. Blood flow to grossly atretic follicles did not differ significantly from that to non-atretic follicles of an equivalent size. Change in blood flow do not appear to initiate or control the activation, steriodogenic function or atresia of follicles.


Effect of size and sex ration of litter on the sexual maturation of female mice.

The relationships between the size and sex composition of the litter and puberty in female mice were investigated. Both factors significantly affected the age of first vaginal oestrus; litter size accounted for 22% of the total variation in age at puberty, and litter sex composition for 8%. When no males or only one male was present in litters of 10 young the females matured significantly earlier than when two or more males were present.


Reproduction in rats and mice isoimmunized with homogenates of ovary or testis with epididymis, or sperm suspensions.

Female rats and mice were injected with homologous spermatozoa without adjuvant, or with homogenates of ovary or testis+ epididymis in adjuvant. Ovulation was not inhibited and the numbers of eggs released were normal. Fertilization rates were significantly reduced in the mouse by all tissues injected and in the rat by injection of ovarian homogenate only. Fracture of and a weak precipitate on the zona pellucida of the eggs were observed in the mouse but not in the rat after injection of ovarian homogenates. The number of pregnancies and litter sizes were reduced following the first and second mating after treatment in the mouse but not in the rat. When eggs with or without follicular cells were examined in vitro, fertilizability was very low if the donor had been treated with ovarian homogenate, better with testis+epididymis homogenate and best after treatment with sperm suspension. Failure of fertilization appeared to be the major cause of infertility after such treatments.


Prediction of fetal sex in cattle by testosterone levels in allantoic fluid.

The testosterone concentration in allantoic fluid between 90 and 150 days of gestation in cattle can be used to determine the fetal sex; values were 442 +/- 20-3 (S.E.M.) pg testosterone/ml for males fetuses and 215 +/- 8-2 pg/ml for female fetuses.


Heterotopic pregnancy following clomid treatment.

A case of concomitant intra- and extrauterine pregnancy after clomiphene treatment is reported. The incidence of multiple ovulations and multiple pregnancies is higher after induced ovulation. The increased likelihood of combined intra- and extrauterine pregnancy in patients with a higher rate of fraternal twinning is discussed.


Termination of late first-trimester and early second-trimester gestations with intramuscular 15 (S)-15-methyl-prostaglandin F2alpha.

15 (S)-15-methyl-prostaglandin F2alpha (15 methyl PGF2alpha) was administered intramuscularly to 222 healthy women seeking abortion of gestations between eight and 22 weeks. Ninety-seven per-cent of these pregnancies were terminated with a mean time to abortion of 16.5 (SD 6.78) hours. Gastrointestinal side effects occurred in more than 90% of the patients while temperature elevations greater than 100.6 degrees F were noted in 22% of cases. No other significant complications were encountered. Transabdominal intra-amniotic pressure monitoring indicated the need to administer drug at two-hour intervals.


Endometriosis in laparotomy scars.

Endometriosis of laparotomy scars has been described as rare. Interruption of pregnancy by abdominal hysterotomy affords an opportunity for its development. The two cases which are reported were admitted to the hospital within one month, which suggests that this diagnosis may not be rare.


Renal carbuncle: unusual cause of postpartum febrile morbidity.

Obstetricians should be alert for medical problems disguised as postpartum complications. A patient with renal carbuncle which presented as postoperative fever is described. The lesion was treated by percutaneous drainage. A brief summary of the differential diagnosis of renal abscess is presented.


Location and removal of misplaced or embedded intrauterine devices by hysteroscopy.

Hysterosopic examinations were performed on 22 patients for IUDs suspected of being misplaced or embedded. Devices included 11 Majzlin springs, nine Lippes loops and two other types. Preoperative preparation and hysteroscopic techniques for extraction are described.


The Hysteroflator 1000S.

A new electrical apparatus for CO2 hysteroscopy is described. It adds an electric suction pump to the insufflator to fix the cervical cup securely and maintain the seal indefinitely. The apparatus was used in 257 patients without any untoward effects.


Synthesis and some pharmacological properties of [1-(L-2-hydroxy-3-mercaptopropanoic acid), 4-threonine]oxytocin (hydroxy [4-thr]oxytocin), a peptide with strikingly high oxytocic potency and of [1-(L-2-hydroxy-3-mercaptopropanoic acid)]oxytocin (hydroxy-oxytocin).

[1-(L-2-Hydroxy-3-mercaptopropanoic acid), 4-threonine]oxytocin (hydroxy[4-Thr]oxytocin) and [1-(l-2-hydroxy-3-mercaptopropanoic acid)]oxytocin (hydroxy-oxytocin) were synthesized by a combination of solid phase and classical methods of peptide synthesis. Protected octapeptides were synthesized by the solid-phase method and 1 + 8 couplings in solution were then employed to furnish the required key protected intermediates. Hydroxy[4-Thr]oxytocin has oxytocic potency as measured in the rat uterus suspended in a Mg2+-free solution, of about 4200 units/mg, eight times the potency of oxytocin, while its antidiuretic potency is approximately equal to that of oxytocin. It thus exhibits a significantly favorable oxytocic-antidiuretic sleectivity. Hydroxy-oxytocin has an oxytocic potency of approximatels 1300 units/mt, 2.5 times that of oxytocin. Threonine substitution in hydroxy-oxytocin has thus caused a significant enhancement in both oxytocic potency and selectivity. The enhancement in oxytocic potency of these two peptides relative to oxytocin and [4-Thr]oxytocin appears to correlate with their lipophilic characteristics, suggesting a significant role of lipophilicity in the interplay of oxytocin-like peptides with oxytocic receptors.


The fine structure of the sperm of a holothurian and an ophiuroid.

The fine structure of the mature sperm of the holothurian, Cucumaria miniata, and the ophiuroid, Ophiopholis aculeata, is described with particular reference to their acrosomal and centriolar satellite complexes, and compared to the sperm of other echinoderms. In Cucumaria, the acrosome is in the form of a diffuse acrosomal vesicle. It is unusual in that it apparently lacks an acrosomal membrane. A membrane separating the acrosomal vesicle from the periacrosomal material may not be equivalent to a typical inner acrosomal membrane. In Ophiopholis, the acrosome is dense, with some internal substructure, and is enclosed by a complete acrosomal membrane. In both species, the acrosome is partially surrounded by an amorphous periacrosomal mass. There is a notable absence of a subacrosomal depression and associated structures as found in other echinoderm sperm. The centriolar satellite complex (CSC) is essentially identical in both species. A reconstruction of the CSC is presented. The CSC consists of nine satellites radiating angularly from the distal centriole, each bifurcating at a dense node before inserting on a marginal ring containing circumferential microtubules. The ring is probably a cytoskeletal element. Immediately below the satellites are nine Y-shaped connectives, connecting each of the axonemal alpha doublets to the flagellar membrane.


Studies on the formation of germ cells in a compound ascidian Botryllus primigenus Oka.

Gametogenesis of a compound ascidian Botryllus primigenus was studied histologically. On either side of the zooid (stage 9), in the gonadal space between the epidermis and the atrial epithelium, either a single testis or a complex of an egg follicle and a testis can be formed. The egg follicle consists of a single ovum (occasionally two ova) and its accessory cells and is connected with the atrial epithelium by the follicle stalk. The eff follicle is always accompanied by the brood pouch, a diverticulum of the atrial cavity. The testis is equipped with a vestigial spermiduct and is attached to the atrial epithelium. Buds of stage 8 comprise, besides the developing testes and egg follicles, loose aggregations of hemoblasts and oocytes of early developmental stages, which are already accompanied by primary follicular cells. Both the oocytes and the primary follicular cells seem to arise from the hemoblasts. The young oocytes are isolated in the gonadal space of the buds and are transferred to buds of the succeeding generations until they finally mature. In the bud of stage 3, a compact mass of cells appears, attaching to the inner vesicle on either side of the body. It is derived from the hemoblasts lodged there in the preceding generation and presumably also from the circulating hemoblasts. When the cell mass receives a large oocyte derived from the preceding generation, part of the cell mass differentiates into egg envelopes, forming an egg follicle, and a follicle stalk and the remainder into a testis. When the cell mass receives no oocyte, it differentiates as a whole into a testis. In the egg follicle thus formed, the outer and inner follicular cells increase in number by mitotic division. Subsequently, initial test cells are derived from the inner follicle by migration across the developing chorion; then they increase in number by mitosis. In the testis, meiosis and spermiogenesis take place.


Ovariectomy alone or in combination with dexamethasone in patients with advanced breast cancer and high levels of testosterone excretion.

The urinary testosterone levels of 40 premenopausal and postmenopausal patients with advanced breast carcinoma were assayed before and after ovariectomies. Histologic examination of the ovaries of 25 patients whose hormone levels were above normal revealed interstitial cell hyperplasia. Two months after the operations, the amounts of testosterone excreted by the womaen decreased significantly. We considered this as indirect evidence that the ovaries contributed to androgen synthesis. However, 15 patients had normal hormone excretion values that did not change 2 months postoperatively. Testosterone levels for 6 of the women 5 months after surgery were higher than at the previous testing. We surmised that the increased gonadotropic activity that followed ovariectomy migh have stimulated adrenal androgen secretion. To elucidate this point, we gave injections of 15,000 IU human chorionic gonadotropin (HCG) to breast cancer patients who were free of any recurrence and, 3 or 24 months previously, had undergone prophylactic ovariectomies (5 and 4 patients, respectively). In 5 of these women, the amount of testosterone excreted increased significantly after HGG was administered. We inhibited adrenal androgen secretion in 11 patients, whose presurgical urinary testosterone levels were above normal. After their ovaries were removed, these women received 1.5 mg dexamethaxone (DXM) daily for 30 days at a time, after which the amount of testosterone excreted was evaluated and, if the level was above 5.0 mug/24 hours, therapy was repeated for another 30 days. Of the patients who showed objective remission after ovarietomies, there were 4 (26.3%) of 15 who had normal androgen excretion levels; 8 (57.1%) of 14 had increased amounts and were treated by surgery alone; and 10 (90.9%) of 11 received a combination of ovariectomies and DXM therapy. The duration of regression was the highest in the latter group.


Phase I study of a five-day intermittent schedule for 1,2:5,6-dianhydrogalactitol (NSC-132313).

Because 1,2:5,6-dianhydrogalactitol (NSC-132313 (DAG; the main conversion reaction product of the treatment of dibromodulcitol by mild akali or human serum) showed considerable antitumor activity in various mouse and rat tumor systems, a phase I study in 50 patients was conducted with five daily iv treatments repeated every 6 weeks. Thrombocytopenia was the dose-limiting toxicity. At a dose of 40 mg/m2/day for 5 days, the median platelet nadir was 31,000/mm3 and occurred on day 20; the plate count returned to normal within 8 days. At the same dose, the median white blood cell (WBC) nadir was 2,300/mm3 also on day 20-, the WBC count returned to normal within 7 days. Anemia, nausea, and vomiting were usually mild to moderate. No renal, hepatic, central nervous system, cardiac, or pulmonary toxicity was identified. Antitumor effects of DAG were observed in patients with renal, bladder, and small-cell lung cancers. An iv dose of 20-30 mg/m2/day for 5 consecutive days, repeated every 5-6 weeks, was recommended for phase II studies.


Patterns of invasive growth in vitro. Human decidua graviditatis confronted with established human cell lines and primary human explants.

Human decidua graviditatis was used as a receptor tissue to demonstrate invasive properties of human cells in vitro. In the confrontation between decidua and established cell lines of tumors and of normal origin as well as primary explanted cells, we noted differences in the cells' interactions with decidual tissue. The two original tumor cell lines, HeLa (carcinoma) and AFi (sarcoma), and a spontaneously transformed lymphoblastoid cell line showed aggressiveness, i.e., invasion, rapid proliferation, injury, and destruction of the decidual tissue. Strain HeLa S3 and two established cell lines, FL (amnion) and Girardi Heart, which are regarded as transformed cells because of their increased mitotic rate and polyploidy, exhibited various degrees of aggressiveness but did not invade. Freshly explanted fetal lung and endometrium caused no injury to the receptor tissue and were included in the decidual tissue.


Determination of adhesive properties of variant metastatic melanoma cells to BALB/3T3 cells and their virus-transformed derivatives by a monolayer attachment assay.

The hypothesis that abnormalities in intercellular adhesion are a property of metastatic tumors was examined in vitro with B16 melanoma variants that were selected in vivo for increased metastatic behavior. The adhesive characteristics of low (B16-F1), intermediate (B16-F5), and high (B16-F10) metastatic lines were determined by quantitative adhesion assays that measured the rate and degree of attachment of single cells to confluent monolayers of melanoma, BALB/3T3, or virus-transformed 3T3 cells. Intercellular adhesions were monitored by loss of single cells from suspension and adherence of intraperitoneally grown 125I-5-iodo-2'-deoxyuridine-labeled cells to the monolayers, and were affected by time, temperature, and serum concentration. Although there was little difference in adhesive properties between the untransformed and transformed 3T3 cell lines, the more metastatic melanoma variants exhibited higher relative rates and extents of homotypic and heterotypic monolayer attachment compared with lower metastatic lines (B16-F10 greater than B16-F5 greater than B16-F1). The correlation between in vivo and in vitro tumor cell adhesive properties and metastasis was discussed.


Neovascularization induced by intraocular xenografts of normal, preneoplastic, and neoplastic mouse mammary tissues.

The capacity to induce neovascularization was compared in normal, preneoplastic, and neoplastic mouse mammary tumors from strains with a high incidence of tumors (C3H, C3H-Avy, C3H-AvyfB) by implantation of small biopsy fragments on the iris surface in New Zealand White rabbits. Proliferation of iris blood vessels was studied by: 1) direct, in vivo slit-lamp stereomicroscopy and fluorescein angiography; 2) filling of the microvasculature with colloidal carbon; and 3) histologic examination. Ninety percent of mammary tumor implants elicited iris neovascularization after 48-72 hours, regardless of their histologic classification or the presence or absence of mammary tumor virus. Corticosteroid treatment reduced immediate postoperative inflammation (12-36 hr) but did not abolish subsequent growth of new vessels. Necrotic tumor fragments failed to elicit any neovascular response. In contrast, only 6% of normal tissues from resting mammary glands caused any vasoproliferation. Hormone-stimulated mammary tissues from pregnant and lactating mice exhibited a transient neovascular capacity that was lost during postweaning involution. Of the implants from premalignant hyperplastic alveolar nodules (HAN's), 30% produced a pattern of vessel growth similar to that of tumor implants. D-1 line (HAN outgrowth) tissues, which have a predicted low incidence of tumors, induced significantly fewer neovascular responses (P less than 0.002) than morphologically and biochemically similar D-2 line tissues, which have a predicted high incidence of tumors. These data suggest that the capacity to induce neovascularization is acquired during malignant progression of mouse mammary tissues; therefore, demonstration of this property may be useful in the identification of those intermediate populations most at risk for neoplastic transformation.


Human gastric choriocarcinoma serially transplanted in nude mice.

A gastric choriocarcinoma cell line capable of producing human chorionic gonadotropin (HCG) was studied by sc transplantation and serial passages into nude mice fed under specific pathogen-free (SPF) conditions. The tumor-take rate at the serial heterotransplantation in SPF mice was high (greater than 95%), in contrast with a low rate in coventional animals (35%). The restoration of morphology and function of the original neoplasm were exclusively verified in SPF mice. Multiple lung metastases were found in 2 animals. The production, storage, and excretion of HCG by tumor cells were confirmed by its high content in serum and cystic fluid in tumors and by its intracellular localization. The tumor cells also contained a specific placental alkaline phosphatase in their membranes. The cells were various trophoblastic types ranging from primitive cytotrophoblasts to typical syncytial cells. The hormone effects of the tumor on sex organs of tumor-bearing animals were evident.


Inhibition and reversal by beta-retinoic acid of hyperplasia induced in cultured mouse prostate tissue by 3-methylcholanthrene or N-methyl-N'-nitro-N-nitrosoguanidine.

The effect of beta-retinoic acid (RA) on carcinogen-induced hyperplasia was studied in organ cultures of mouse prostate gland. 3-Methylcholanthrene (MCA), requiring metabolic activation, or N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), not requiring activation, were used to induce hyperplastic changes. Treatment of cultures with MCA or MNNG stimulated cell proliferation and caused the alveolar epithelium to become hyperplastic. The development of this hyperplasia was inhibited when RA was added simultaneously with MCA or MNNG. However, RA had no significant effect on cell proliferation in untreated control cultures. Elimination of carcinogen from the hyperplastic cultures after 8 days of treatment did not reverse hyperplasia of the alveolar epithelium. When the withdrawal of MCA or MNNG was followed by treatment of the cultures with RA, hyperplasia was markedly reversed within 96 hours. Thus RA actively inhibited and reversed the effect of MCA and MNNG, two carcinogens that may have different mechanisms of action.


Azoxymethane-induced liver hemangiosarcomas in inbred strain-2 guinea pigs.

Intrarectal administration of 0.5 ml of a 0.2% solution of azoxymethane twice weekly for 33 weeks to female inbred strain-2 guinea pigs specifically induced multiple liver hemangiosarcomas in 15 of 16 animals 32-54 weeks after the first treatment. No neoplasms and preneoplastic changes were found in the large intestine.


Effects of delta9-tetrahydrocannabinol in Lewis lung adenocarcinoma cells in tissue culture.

We found a dose-related decrease in DNA synthesis in transformed cell cultures treated with delta9-tetrahydrocannabinol (delta9-THC). The decrease, observed over a 4-hour period, was not accompanied by a change in the radioactive precursor pool as compared to that of control culture. The distribution of labeled products clearly differed from that observed after treatment with cytosine arabinoside. delta9-THC inhibited DNA synthesis at some point beyond the uptake of 3H-thymidine.


Effects of diethylnitrosamine and cigarette smoke on hamsters.

One hundred two 2-month-old male Syrian hamsters received 12 weekly sc injections of 0.25 mg diethylnitrosamine (DEN). Half the animals were also exposed for 10 minutes to cigarette smoke in Hamburg II-type smoking machines, three times per day, 5 days per week, for their life-span. The other half received sham exposure. Two additional groups of 51 hamsters each, serving as controls, received either only smoke or sham smoke exposures. The DEN treatment resulted in a significant increase (P less than 0.01) in epithelial lesions of the larynx, including laryngeal papillomas. Cigarette smoke inhalation had a significant (P less than 0.01) potentiating effect on the incidence of these lesions. We did not observe the high incidence of tumors of the nasal cavity, trachea, and the lower respiratory tract, or a significant development of malignant tumors, reported by several other investigators. The life-spans of the animals were unaffected by DEN and significantly increased (P less than 0.01) by exposure to smoke. The smoke-exposed groups had significantly lower mean body weights than their sham smoke-exposed cohorts.


Tumor-specific immunogenicity induced by chemical modification.

C57BL/6J spleen cells were immunized in vitro against cells from a syngeneic 3-methylcholanthrene-induced sarcoma, either modified with diazotized sulfanilic acid or left unmodified. The spleen cells were harvested after 5 days and run in a short-term 51Cr-release assay against unmodified tumor cells as targets. The spleen cells sensitized against the modified but not the unmodified tumor cells were cytotoxic for the unmodified tumor cell targets but not for two other syngeneic tumor cell lines or syngeneic spleen cells, thus indicating the production of tumor-specific cytotoxicity.


External stabilization of a replanted upper extremity: case report.

A case replantation of the upper extremity is presented using external fixation as the method to achieve bony stability. Although internal fixation may remain the method of choice for stabilization of a replanted limb, external fixation can also provide bony stability where more conventional methods are not applicable.


Polypeptides of mumps virus.

Mumps virus was propagated in the extra-embryonic fluids of embryonated chicken eggs and was labeled by cionjection of radioactively labeled amino acids. The virus was purified by density gradient centrifugation, and its polypeptides were analyzed by polyarylamide gel electrophoresis. The virus was found to be composed of six polypeptides, ranging in size from 40,000 to 64,000 daltons. Viral proteins 1 and 3 were the glycoproteins of the virons. When the virus particle was treated with noniontic detergents, a small fraction of these glycoproteins could be released into the supernatant. After treatment with nonionic detergents in high salt and alkaline conditions, more of the surface glycoproteins were removed. This treatment also released the smallest viral polypeptide from the virion. The glycoproteins were separated using an affinity chromatographic column of agarose-fetuin. The heavier glycoprotein, viral protein 1, was found to contain both the neuraminidase and hemagglutinating activity. The two glycoproteins were tested for their ability to react in complement-fixing tests with mumps antisera. Only the heavier glycoprotein reacted with antisera possessing both anti-S and anti-V activity. Neither glycoprotein reacted with antisera specific for the S antigen. Thus, it was concluded that this glycoprotein corresponds to the classical V antigen of mumps virus.


Growth and genetic stability of the ts-1 mutant of respiratory syncytial virus at restrictive temperatures.

An in vitro study was performed to define in greater detail those factors which favored the growth of the ts-1 mutant of respiratory syncytial virus under restrictive conditions and the emergence of genetically altered virus with decreased temperature sensitivity. Replication of ts-1 occurred at each of the restrictive temperatures of 37, 38, and 39 C, even through plaque formation was not observed. The level of virus growth under restrictive conditions was inversely related to the incubation temperature and directly related to the multiplicity of infection. These relationships appeared to reflect the effect of restrictive temperature in reducing the quantity of virus produced and released from an infected cell. Under restrictive conditions the production of genetically altered virus which exhibited reduced temperature sensitivity was directly related to the multiplicity of infection and inversely related to temperature. Production of genetically altered virus was not observed under permissive conditions.


Mason-Pfizer virus RNA genome: relationship to the RNA of morphologically similar isolates and other oncornaviruses.

The 60-70S RNA of Mason-Pfizer virus (MPV) was iodinated in vitro and used in both direct and competitive molecular hybridization studies. MPV proviral sequences are present at a frequency of approximately one to two copies per haploid genome in the DNA of experimentally infected human cells. By nucleic acid competition hybridization, MPV RNA was found to be indistinguishable from the RNA of a virus (X381) isolated from a rhesus mammary gland and from RNA isolated from the cytoplasm of AO cells (Parks et al., 1973) and HeLa cells (Gelderblom et al., 1974), both previously reported to produce MPV-related particles. No homology was observed, however, between MPV RNA and the RNA, or the DNA, from two clones of HeLa cells obtained from the American Type Culture Collection. Hybridization of MPV 60-70S RNA to the DNA of normal tissues of humans and to the DNA of 11 other species revealed that MPV is not an endogenous virus of any of these species. Competition hybridization revealed no detectable sequence homology between the RNA of MPV and the RNAs of simian sarcoma virus, murine mammary tumor virus, murine leukemia virus, BUdR-induced guinea pig virus, or avian myeloblastosis virus. These nucleic acid studies substantiate previous ultrastructural and immunological findings that MPV and morphologically similar isolates constitute a distinct group of oncornavirus.


Endogenous oncornaviral DNA sequences: evidence for two classes of viral DNA sequences in guinea pig cells.

The nature of the endogenous viral DNA sequences in guinea pig cells was studied by hybridization. A segment of the viral RNA (r-VRNA) hybridizing to abundant (or reiterated) DNA sequences (R-VDNA) was isolated by recycling to a Cot of 300. The hybridization of the recycled VRNA, as well as the total VRNA, was followed by determining their kinetics and by Wetmur-Davidson analysis. The kinetics of hybridization of total VRNA were complex, did not follow a second-order kinetics, and revealed two slopes by Wetmur-Davidson analysis. The recycled RNA, on the other hand, had a second-order reaction rate expected of the hybridization between a single species of RNA and DNA sequences and yielded a single straight line in a Wetmur-Davidson plot. The Cot1/2 and slope of the recycled r-VRNA was almost identical to that of the abundant VDNA sequences obtained from the hybridization data of the total VRNA. Guinea pig 28S rRNA with or without recycling was used in monitoring hybridization rate. The kinetics of hybridization of 28S RNA followed a second-order reaction and produced a single straight line by Wetmur-Davidson plot, with a second-order reassociation rate constant of 9.6 x 10(-3) liters/mol-s, a Cot1/2 of 104 mol-s/liter, and reiteration frequency of 146. There was no difference in the kinetics of hybridization of 28S RNA before and after recycling. These experiments showed that guinea pig cells contain two classes of VDNA sequences. (i) R-VDNA sequences with a second-order reassociation rate constant of 8.2 x 10(-4) liters/mol-s, a Cot1/2 of 1,219 mol-s/liter, and a reiteration frequency of 12 represent 37.5% of the viral genome. (ii) Unique VDNA sequences with a second-order reassociation rate constant of 1.2 x 10(-4) liters/mol-s, a Cot1/2 of 7,692 mol-s/liter, and a reiteration frequency of 2 represent 62.5% of the viral genome.


Biochemistry of DNA-defective mutants of bacteriophage T4. VI. Biological functions of gene 42.

Bacteriophage T4 gene 1 and 42 amber mutants (defective in deoxynucleoside monophosphate kinase and deoxycytidylate hydroxymethylase, respectively) are able to synthesize DNA in cell-free lysates prepared as described by Barry and Alberts (1972), in contrast to their inabliity to do so in plasmolyzed and toluenized cell systems. Addition of extracts containing an active gene 1 or 42 product has no effect on synthesis in lysates defective in the respective gene. Thus, if these enzymes do play additional direct roles in replication, these roles are not manifest in the lysed-cell system. The gene 42 mutant am N122/m, a double mutant bearing an additional defect in DNA polymerase, is unable to synthesize DNA in these lysates. This inability is overcome by addition of extracts containing an active T4 DNA polymerase. m is a leaky amber mutation which reduces DNA polymerase activity to a very low level. However, this level is high enough to allow positive genetic complementation tests with gene 43 mutants. Two other gene 42 amber mutants contain additional defects: am 269 induces only half the normal level of DNA polymerase, and am C87 fails to induce a detectable level of thymidylate synthetase. These defects do not result from pleiotropic effects of the gene 42 mutations. In plasmolyzed cells, temperature-sensitive gene 42 mutants fail to synthesize DNA under conditions where replication forks and 5-hydroxymethyl-dCTP are present. This supports the idea that the gene 42 protein is directly involved in DNA synthesis.


Influenza virus RNA's in the cytoplasm of chicken embryo cells treated with 3'-deoxyadenosine.

3'-Deoxyadenosine (75 to 100 mug/ml) permitted analysis of the cytoplasmic influencza virus-specific RNAs synthesized early in the replicative cycle-a phase that has hitherto been obscurred by host cell RNA synthesis. In addition, late in the cycle (6 to 8) complementary virus-specific RNA's were the predominantly labeled species, suggesting that higher concentrations of 3'-deoxyadenosine selectively inhibit influenza viral genome replication.


Advanced renal cell carcinoma: treatment with xenogeneic immune ribonucleic acid and appropriate surgical resection.

Herein we describe the first clinical treatment of renal cell carcinoma in humans with xenogeneic immune ribonucleic acid. Twelve patients with advanced renal cell carcinoma have been treated by appropriate operations to remove tumor bulk followed by specific passive immunotherapy. Xenogeneic specific immune ribonucleic acid was prepared from the spleen of normal sheep that had received 4 weekly injections of a homogenate of renal cell carcinoma. Results indicate that 1) xenogeneic specific immune ribonucleic acid can safely be given to humans without local or systemic toxicity, 2) there is a suggestion of clinical benefit, since only 2 patients have had progression of known metastases during treatment with immune ribonucleic acid and 3) xenogeneic immune ribonucleic acid can enhance the immune response to renal cell carcinoma, as demonstrated by in vitro lymphocytoxicity tests.


Serum immunoglobulins in genitourinary malignancies.

Serum concentrations of immunoglobulins G, A and M (IgG, Ig A and IgM) were measured by immunodiffusion in a control population of 58 patients and in 165 patients with cancer of the prostate, bladder, kidney or testis. Tumor patients were ranked according to clinical staging. In a smaller group of patients serum levels of IgE were measured by radioimmunoassay. IgG, IgA and IgM concentrations did not correlate with staging or the presence of malignancy. However, a statistically significant correlation between increased IgE levels and advanced bladder cancer (staged D1 and D2) stimulated us to study more patients with bladder tumors. In a subsequent group of 22 bladder tumor patients this trend could not be reproduced. IgE levels did not correlate with a history of allergy in any of the patients tested. Concentrations of immunoglobulins G, A, M and E seem to be unchanged in genitourinary malignancies, although their functional role here has not been assessed qualitatively.


Prognosis for resolution of moderate, primary reflux in girls.

Detailed cystoscopic grading of ureteral orifices has proved unnecessary in our experience for determining the prognosis of moderate, primary reflux in girls. Medially placed orifices strongly predict spontaneous resolution with preventive medication, while radiographic pyelonephritic scarring and/or grade III reflux strongly predict failure to resolve. While laterally placed ureteral orifices and low volume reflux suggest a need for operation, they are prognostically less useful, particularly when those with grade III reflux are excluded. Cystitis cystica and unrelated anomalies seem to be associated with the ultimate need for operation but are of uncertain significance.