Effects of cholinoceptor antagonists on the suckling-induced and experimentally evoked release of oxytocin.

1 In the anaesthetized lactating rat, the suckling of the young causes the regular release (about every 7 min) of brief pulses of oxytocin (0.5 to 1.0 mu), which each produce a single transient increase in intramammary pressure.2 The effects of several cholinoceptor antagonists were studied in relation to this natural reflex, and also the release of oxytocin evoked by the intraventricular injection of cholinomimetics.3 Reflex milk ejection was blocked by the nicotinic antagonists mecamylamine and hexamethonium, and the inhibition was dose-dependent (ED(50) of 1 mg/kg i.v. and 5 mg/kg i.v., respectively). Despite the use of high doses, the muscarinic antagonists atropine (200 mg/kg), hyoscine (90 mg/kg) and benzhexol (30 mg/kg) all failed to prevent the reflex release of oxytocin.4 Acetylcholine (20 to 100 mug), bethanechol (0.2 to 4.0 mug) and carbachol (0.01 to 0.2 mug) injected into the cerebral ventricals stimulated a sustained release of oxytocin, which produced multiple increases in intramammary pressure. Nicotine (200 mug) was relatively ineffective by this route.5 The release of oxytocin by intraventricular bethanechol or carbachol was abolished by atropine (0.1 to 1.0 mg/kg) but not by mecamylamine (5 mg/kg) or hexamethonium (5 mg/kg).6 None of the antagonists used significantly affected either the release of oxytocin following electrical stimulation of the neurohypophysis or the mammary sensitivity to endogenous or exogenous oxytocin.7 The results suggest that the neural pathway controlling the reflex release of oxytocin during suckling in the rat contains a cholinergic component, which acts through nicotinic receptors. A second cholinergic pathway, of the muscarinic type, may also exist. The role of these two pathways is discussed.


Budgerigar-fancier's lung: the commonest variety of allergic alveolitis in Britain.

A questionnaire survey of 1005 consecutive attenders at four outpatient clinics yielded 117 (12%) budgerigar fanciers (exposed to budgerigars- known in North America as parakeets-for at least three months) and 296 (29%) former fanciers. Twnety had precipitins to budgerigar serum or droppings or both, and 10 of these together with 39 precipitin-negative patients reported undue breathlessness on exertion during exposure to buderigars. These 59 patients were investigated further, seven completing a series of inhalation provocation tests with budgerigar antigens designed to confirm or exclude budgerigar-fancier's lung (BrFL). Typical positive responses were obtained from four current and one former fancier. The prevalence of confirmed BrFL among the 11n current budgerigar fanciers was 3.4% (four cases). This was biased, however, by the inclusion of one patient whose attendance at the surveyed clinic was attributable to the disease. With the exclusion of this patient, confidence limits suggested that the true prevalence of BrFL among current budgerigar fanciers in the general population lies between 0.5% and 7.5%, which is similar to the prevalence of farmer's lung in farm workers. In view of the enormous population at risk, however, this implies that BrFL rather than farmer's lung is by far the commonest type of allergic alveolitis in Britain.


Absence of glucoprivic feeding after stress suggest impairment of noradrenergic neuron function.

Feeding in response to 2-deoxy-D-glucose (2DG), a quantifiable behavior which appears to depend on noradrenergic (NE) neuron function, was used in these experiments to evaluate the functional capabilities of NE neurons after stress exposure. Depletion of hypothalamic NE after footshock or hypothermic stress was directly correlated with impairment of glucoprivic feeding. When NE depletion was prevented by prior exposure to chronic stress, no impairment of feeding was observed. After hypothermic stress, repletion of NE proceeded more rapidly in the telencephalon than in the hypothalamus and reappearance of a normal feeding response precisely paralleled the time course of repletion in the hypothalamus. Drinking in response to cell dehydration, a behavior not directly dependent on brain catecholamines, was not impaired after either footshock or hypothermic stress, despite similar NE depletions. Presence of a normal drinking response assured that deficits observed in the 2DG test were not due to nonspecific behavioral suppression resulting from stress. These data suggest that NE neuron function may be impaired or temporarily abolished after severe stress exposure. In addition, these results demonstrate that behavioral pathology need not be the result of massive neurotransmitter depletion but may result from relatively subtle alterations of specific neurotransmitter pools.


Nocturnal rotation in normal rats: correlation with amphetamine-induced rotation and effects of nigrostriatal lesions.

Normal unoperated rats were observed to rotate (turn in circles) at night. For most (91.7%) rats, the preferred direction of rotation was consistent across hours and days and was the same as the direction of rotation elicited by D-amphetamine (1.0 mg/kg) during the day. The magnitudes of nocturnal and D-amphetamine-induced rotation were also highly correlated. After rats showed stable diurnal patterns of rotation, unilateral lesions were made in either the substantia nigra, the nigrostriatal bundle or the caudate nucleus. All lesions produced transient contralateral rotation within the first 24-48 h after surgery. The time-course of this contralateral rotation was more prolonged after nigral lesions than after nigrostriatal bundle lesions and least after caudate lesions, suggesting that the duration of a degeneration release of dopamine is proportional to the length of the degenerating axon. Lesion size was correlated with the intensity of contralateral rotation but not with the time-course. At each rostralcaudal level, the magnitude of contralateral rotation was greater if the lesion was in the side of the brain opposite to the preoperative direction of rotation than if in the same side. By three days after surgery, all rats returned to a mostly normal diurnal cycle with the direction of rotation now being ipsilateral to the lesion. D-Amphetamine potentiated the ipsilateral rotation, though rats with lesions in the same side of the brain as the preoperative direction of rotation had larger drug responses than rats with similar lesions in the opposite side of the brain. By one month after surgery, the direction of spontaneous rotation of most rats had returned to the preoperative direction. As at all other times, the magnitude of rotation was, in part, dependent on the side of the lesion with respect to the preoperative bias. It is suggested that, following a unilateral lesion, compensatory processes occur to a greater extent if the lesion is in the normally more active side of the brain.


Effect of p-chloroamphetamine and 5,7-dihydroxytryptamine on rotation and dopamine turnover.

Injections of p-chloroamphetamine (PCA) or 5,7-dihydroxytryptamine (DHT) (after pretreatment with desmethylimipramine) into the median raphe nucleus (MRN) caused depletions of 5-HT and 5-HIAA in the cortex and striatum, and a decrease of cortical 5-HT uptake without affecting NE uptake. Unilateral injections of these neurotoxins into the MRN caused contralateral rotation, which was blocked by haloperidol. The size of the lesion correlated with the rate of rotation and the decrease in 5-HT turnover in the cortex. We also found a significant correlation between the rate of rotation and the decrease in cortical 5-HT turnover, and the increase in striatal DA turnover. Moreover, there was a significant correlation between the decrease in cortical 5-HT turnover and the increase in striatal DA turnover. It was found that injections of DHT into the SN produced similar behavioral and biochemical changes as did the MRN lesions. In this model, amphetamine and apomorphine produce turning in the same direction, whereas they have opposite effects after lesions of the nigrostriatal pathway where postsynaptic DA supersensitivity occurs. Presynaptic changes appear to determine turning in this model. An inhibitory role of the serotonergic MRN on the dopaminergic nigrostriatal pathway mediated via the substantia nigra (SN) is suggested.


Metacarpal morphometry in monozygotic dizygotic elderly twins.

The relative importance of genetic factors in the pathogenesis of age related bone loss has been investigated in a study involving 17 monozygotic (MZ) and 8 dizygotic (DZ) pairs of twins aged 64 to 75 years. Radiographic morphometry was performed at the midpoints of the 2nd, 3rd and 4th metacarpals of both hands and the mean total and cortical widths were evaluated. The heritability, h2, was calculated as the difference between the intrapair variances in same sexed DZ and MZ pairs divided by the intrapair variance in DZ pairs. The mean intrapair variance of both total and cortical width was found to be four to five times higher in DZ than in MZ pairs. The differences are highly significant with an h2 value between 0.7 and 0.8, indicating a predominant genetic influence. It is stressed that this result applies only to the population from which the twin sample was drawn.


Purification and properties of phosphatidylglycerophosphate synthetase from mammalian liver mitochondria.

The enzyme which catalyzes the synthesis of phosphatidylgly cerophosphate from an-glycerol-3-phosphated and cytidine diphosphate diacylglycerol was released from rat or pig liver mitochondrial membranes by extraction with Triton X-100 or Nonidet P-40. The detergent-extracted enzyme, like the activity of intact mitochondria, did not require added cations or lipids. The Triton extracts were fractionated by column chromatography on Bio-Gel A-1.5. The fractions obtained from the columns exhibited little activity in the standard assay system unless divalent cations were included. Additional stimulation (about twofold) was observed in the presence of added phospholipids. The cation requirement of the purified enzyme was relatively nonspecific with Mg2+, Ba2+, or Ca2+ providing maximal activity in the 10mM range. Either Mn2+ or Co2+ were stimulatory at somewhat lower concentrations but higher concentrations were inhibitory. Other cations such as Cd2+, Zn2+,Hg2+, or Cu2+ were ineffective as cofactors, and in the presence of Mg2+ inhibited the reaction at concentrations greater than 0.5 mM. The phospholipik stimulation was obtained specifically with phosphatidylethanolamines from natural or synthetic sources. Other diacylglycerophosphatides or lysophosphatides including lysophosphatidylethanolamine were ineffective.


The defect in transcellular transport of phosphate in the nephron is located in brush-border membranes in X-linked hypophosphatemia (Hyp mouse model).

We purified renal cortex brush-border membranes from mutant hemizygous hypophosphatemic (Hyp/Y) mice and male control (+/Y) littermates. Tenfold purification of mutant and wild-type membranes was obtained. Phosphate enters +/Y brush-border membrane vesicles by a saturable Na+-dependent arsenate-inhibited component and also by a diffusional component observed in the presence of a potassium gradient. Phosphate is not bound or incorporated significantly by mouse brush-border membrane vesicles. Parallel studies with rat renal cortex brush-border membrane vesicles revealed that phosphate and D-glucose transport in rat and mouse vesicles are similar and have the characteristics reported by other workers. Brush-border membrane vesicles prepared from Hyp/Y renal cortex have significant (p less than 0.001) partial loss of phosphate transport on the Na+-dependent arsenate-inhibited component. D-Glucose transport is not affected. Our previous studies reveal that other components of transcellular phosphate flux in kidney are normal. Therefore, we conclude that the mutant gene product in the Hyp mouse is confined to the brush-border membrane. Stability of the X-chromosome in mammalian evolution implied that the same gene product is involved in the classic human disease, familial 'vitamin D 'resistant' X-linked hypophosphatemia.


Establishment of a human plasma cell line in vitro.

A human plasma cell line designated ARH-77 has been established and propagated in culture for the past 2 years. The cells exhibited morphological characteristics of plasma cells under light and electron microscopic examination. An average of 40% cells are positive for immunoglobulin G by direct immunofluorescence, while an immunoglobulin G-specific radioimmunoassay reveals the production of 1.21 X 10(4) ng/10(6) plasma cells. The karyotype is aneuploid with a a modal chromosome number of 45 to 46 and no marker chromosome. Growth kinetics characteristics are: doubling time, 110.4 hr; generation time, 56.4 hr; G1 + G2-phase transit time, 45.5 hr; S-phase transit time, 10.9 hr; growth fraction, 74%; mitotic index, 1.5%; labeling index, 14.3%; and cell loss, 31.0%. Some of the growth kinetics characteristics were markedly similar to the properties displayed in vivo by plasma cells of patients with multiple myeloma and suggest that the cell line might be a useful in vitro model for the study of human myeloma.


Studies on the ultrastructure of the integument of the rotifer Habrotrocha rosa Donner (Aschelminthes).

The integument of the rotifer Habrotrocha rosa Donner is provided with pores and formed by an extrasyncytial cuticle and a syncytial hypodermis. This hypodermis peripherally contains 3 layers of dense cytoplasm and borders the cuticle by an asymmetric cell membrane. The wall of the pores is stiffened proximally like an annulus. The pores lead into cytoplasmic invaginations which are surrounded by vesicles. Close to and also beneath the condensed cytoplasmic layers microbodies are found, which are interpreted as microperoxisomes. Subhypodermal layers of muscles are connected with the cytoplasm of the hypodermis by desmosome-like structures.


Pseudo subaortic stenosis--a catheter-induced artifact.

Three cases are reported of valvular aortic stenosis in which artifactual subvalvular chambers were recorded on pressure tracings. The pressure tracings were obtained during withdrawal of multiholed catheters from the left ventricle to the aortic root. Several recent reports have described the coexistence of valvular and subvalvular aortic stenosis. The cases reported here re-emphasize that a multiholed catheter, which is the type commonly used for retrograde left ventricular entry in a patient with valvular aortic stenosis, is unsatisfactory for the detection of coexisting subvalvular obstruction.


Hormonal regulation of ovarian cellular proliferation.

The steroid hormone estradiol, and the glycoprotein hormones follicle-stimulating hormone (FSH) and luteinizing hormone (LH), are known to be essential for the growth and differentiation of follicles in the ovary. The present study was conducted to determine quantitatively the effects of estradiol, FSH and LH on proliferation of different ovarian cell types (granulosa and theca cells). The immature female hypophysectomized rate sequentially primed with estradiol, FSH and LH was used as the experimental model. Proliferation was assessed by examining changes in total DNA, incorporation of 3H-thymidine into DNA and labeling index in specific cell types. Estradiol and FSH each acted on follicles at different stages of development to stimulate proliferative activity of both granulosa and theca cells. Continued administration of either hormone caused a decrease in the proliferative activity of both cell types. These observations have been interpreted to indicate that estradiol and FSH can each alter the length of the specific phases of the cell cycle. A luteinizing dose of LH caused a cessation of proliferation in luteinizing granulosa cells while stimulating a limited proliferation of theca cells. Absence of the appropriate hormonal stimulus caused both granulosa and theca cells to stop proliferating and the follicles to undergo atresia. These results indicate that, depending upon the state of differentiation of granulosa and theca cells, estradiol, FSH and LH can stimulate or inhibit the ability of these cells to proliferate.


A comparison of two cloned mouse beta-globin genes and their surrounding and intervening sequences.

The BALC/c mouse has two nonallelic beta-globin genes that appear to reside on two different Eco R1 fragments of genomic DNA. We have already cloned one of these fragments and shown that the gene encoded within it is interrupted by at least one large intervening sequence of DNA. We have now cloned and characterized the second beta-globin gene-containing fragment. The coding sequence of its gene is also interrupted by an intervening sequence of DNA that occurs in about the same position, relative to the coding sequence, as does the first. Because some shared features of the structure of these two genes might be responsible for their coordinate expression and the elimination of their intervening sequences, we have compared their surrounding, coding and intervening sequences by restriction endonuclease analysis and by visualization of the heteroduplex structures formed between them. Of the 7000 bp of sequence compared in this way, we find only a few hundred base pairs of homology in addition to the coding sequence. These shared sequences flank the coding sequence and appear to include only those portions of the intervening sequence immediately adjacent to the interrupted structural gene.


Structural perturbations in the plasma membrane during concanavalin A endocytosis.

Endocytosis of Concanavalin A, triggered by its interaction with the surface of cells from a murine plasmocytoma line, is characterised by various steps which can be visualised by cytochemistry and freeze-fracturing. Plasma membrane internalisation is initiated by clustering of Concanavalin A receptors and by formation of intramembraneous particle necklaces, which were observed on fracture faces. Subsequent perturbation of the lipid bilayer precedes the fusion and formation of closed vesicles.


Trypsin treatment which elicits DNA synthesis, removes a high molecular weight glycoprotein from the plasma membrane of sea urchin embryonic cells.

SDS-polyacrylamide gel electrophoresis of plasma membrane proteins from sea urchin embryos at the swimming blastula stage allows the identification of fifteen-seventeen components, five of which are glycoproteins. One of these (molecular weight about 145,000) is almost completely absent in the plasma membrane of trypsin treated cells.


Characteristic proteins systhesized by suspensions of cells isolated from rat gastric mucosa.

The biochemical integrity of rat gastric mucosal cells after isolation with pronase was demonstrated by their synthesis of a wide range of proteins. Approximately 50 bands of pre-existing proteins were detectable by staining on SDS-polyacrylamide electrophoresis gels. Most bands appeared readiolabelled after incubating cells with 14C-leucine or 14C-proline and were detected by autoradiography and fluorography of the gels. Recent synthesis, indicated by the radioactivity of each band, was compared with historical synthesis indicated by the intensity of staining. Densitometric comparisons showed the relative specific radioactivity of certain bands, e.g., one of nominal molecular weight 25000, to be consistently higher than the integrated mean value.


Morphology and 125I-concentration of embryonic chick thyroids cultured in an atmosphere of oxygen.

In vitro culture of whole thyroid glands may lead to loss of differentiation and function. Comparison by histology, histoautoradiography and gammacounting of 125I-concentration, of 12 to 13 days old embryonic chick thyroids, cultured on a gyratory shaker either in an atmosphere of 5% CO2 and air or of 95% O2 and 5% CO2, shows that a high concentration of O2 maintains the morphological and functional integrity of the thyroids for at least 4 days.


Translation "in vitro" of globin mRNA from Xenopus laevis.

RNA isolated from Xenopus laevis reticulocytes and characterized as globin mRNA (Meza et al., 1978) was tested for its capacity to stimulate "in vitro" a wheat germ translation system, and the ability to synthesize a polypeptide. The latter was identified as globin by its electrophoretic mobility and immunoprecipitation with antiglobin antibody.


Characterization of ts13 cells a temperature-sensitive mutant of the G1 phase of the cell cycle.

ts 13 cells are a temperature-sensitive (ts) mutant of BHK cells that are known to arrest in G1 when shifted to the nonpermissive temperature. We have determined the entry into S of ts13 cells in five different growth conditions, namely: 1) quiescent, sparse cultures stimulated to proliferate by serum. 2) Quiescent, dense cultures stimulated by serum. 3) Quiescent, sparse cultures stimulated by trypsinization and replating. 4) Quiescent, dense cultures stimulated by trypsinization and replating. 5) Mitotic cells collected by mitotic detachment. For each different growth condition we have also determined the execution point of the mutant function, i.e. the time at which a shift-up to the nonpermissive temperature no longer prevents the entry of cells into S. The median time of entry into S and the execution point varied in different growth conditions, but the distance between the median execution point and the median time of entry into S was remarkably constant, i.e. 3.2 hr. In addition we have fused ts 13 cells cells with chick erythrocytes and studied the ability of ts13 cells in heterokaryon formation to induce DNA synthesis in chick nuclei. Although ts13 cells can induce DNA synthesis in chick nuclei at the permissive temperature, they fail to do so when fused and stimulated at the nonpermissive temperature of 39.5 degrees C.


Oscillating contractions in protoplasmic strands of Physarum: infared reflexion as a non-invasive registration technique.

An inexpensive electronic device employing a miniaturised infrared reflection detector is described which enables the automatic registration of intrinsic radial contraction activities of protoplasmic strands of Physarum growing on their original substrate. Advantages and shortcoming of the new registration technique are discussed in relation to recent tensiometric techniques applied to protoplasmic strands for analysing the contraction physiology of cytoplasmic actomyosins, the force generating substrate of the contraction automaticity in the Physarum system.


Mutagenesis by N-acetoxy-2-acetyl aminofluorene of chinese hamster V79 cells is unaffected by caffeine.

8-Azaguanine (AZG)- and 6-thioguanine (TG)-resistant cells (mutants) were induced in Chinese hamster V79-4 cells by 0.1--2.5 microgram/ml N-acetoxy-2-acetyle aminofluorine (AcAAF) treatments in the presence of 5% fetal bovine serum (FBS). The frequency of resistant colonies increased from 1 to 47 per 10(5) survivors. The effect of caffeine (50--200 microgram/ml) during the mutagenesis expression period was determined by adding caffeine 1--24 h after AcAAF. The medium was replaced after 48 h exposures so that caffeine was absent during subsequent selection with AZG or TG. No significant change in the AcAAF-induced mutant frequency occurred with any treatment combination although caffeine greatly enhanced the lethality associated with AcAAF treatments. Thus, caffeine interferes with postreplication repair in V79-4 cells without affecting the probability of error of the repair process. These results were obtained with a quantitative mutagenesis assay in which the cells were reseeded prior to selection to achieve maximum expression without interference from metabolic crossfeeding. In contrast, the commonly used in situ assay is subject to serious interference from crossfeeding and yields an artifactual enhancement of AcAAF mutagenesis by caffeine.


The action of AF 2 on cultured hamster and human cells under aerobic and hypoxic conditions.

AF 2 (2-(2-furyl)-3-(5-nitro-furyl)acrylamide) was toxic to Chinese hamster V 79 cells and normal human fibroblasts in aerobic media. However, the toxicity of the drug was increased many times by hypoxia. Similarly, the frequency of AF 2-induced azaguanine- and ouabain-resistant mutants of V 79 cells was much higher in hypoxia than under aerobic conditions. Both hamster V 79 cells and human fibroblasts metabolized AF 2 and other nitrofurans rapidly only under hypoxic conditions. Human fibroblasts were more sensitive to AF 2 both under aerobic conditions and in hypoxia than were V 79 cells under similar conditions. The Chinese hamster cells consistently gave survival curves with marked shoulders while human cells did not. Aerobic cultures of fibroblasts derived from xeroderma pigmentosum (XP) patients were markedly sensitive to AF 2 while fibroblasts from two ataxia telangeictasia patients had normal sensitivity. Under hypoxic conditions the sensitivity of both types of cells was increased but the XP line remained 5--10-fold more sensitive than normal or ataxia cells. These results suggest that the DNA lesions produced by AF 2 may be regarded as similar to those produced by ultraviolet light, at least in terms of their repairability in human cells.


Hemodynamic and metabolic response after abrupt uithdrawal of long-term propranolol.

Because the mechanism of adverse reactions to abrupt withdrawal of propranolol in patients with coronary disease is an enigma, we studied the effect of cessation of propranolol on beta receptor reactivity to catecholamine stimulation. Heart rate and maximum rate of rise of left ventricular pressur (dP/dt max) during isoproterenol infusions and plasma free fatty acids (FFAs) after epinephrine infusions were measured in six conscious dogs before, during and after four weeks of oral propranolol (40 mg p.o. q8h). Rises in heart rate, dP/dt max and FFAs were blocked during propanolol administration. Twenty-four hours after withdrawal from propranolol, heart rate and dP/dt max responses remained significantly attenuated, although FFA responses were at premedication levels. The 72-hour, 96-hour and one week postmedication responses did not differ from premedication values. Thus, partial beta blockade of the heart was still present at 24 hours and no evidence of heightened beta receptor sensitivity was detected on repeated study one week after withdrawal from a long-term, high dose propranolol regimen.


Antibodies to lactalbumin interfere with its radioimmunoassay in human plasma.

Two radioimmunoassays for human lactalbumin have been established using a rabbit antiserum. One assay uses a second antibody to separate bound from free label; the other uses polyethylene glycol to precipitate gamma globulin non-specifically. We have confirmed that about half the normal human population have a substance in their blood which inhibits the binding of lactalbumin to the rabbit antibody. Comparison of the two assays has demonstrated that this material is not lactalbumin but a naturally occurring antibody. We have shown that it is in the IgG fraction of human plasma and is probably a cross-reacting antibody to bovine lactalbumin. None out of fifteen males and fourteen out of fifty eight non-pregnant, non-lactating females had low levels of lactalbumin in the their blood (0.6--2.0 ng/ml). Our assay could not detect a statistically significant difference between normal women and those with either benign breast disease or metastatic mammary carcinoma.


Camptomelic dwarfism associated with XY-gonadal dysgenesis and chromosome anomalies.

We have studied two female newborns with comptomelic dwarfism, XY-gonadal dysgenesis and chromosome anomalies. The preponderance of "females" among the hitherto reported cases of this allegedly autosomal recessive form of lethal drawfism may be due to an increased incidence of an associated XY-gonadal dysgenesis.


[Pasteurella multocida infection after a dog bite (author's transl)].

In a 3-year old girl Pasteurella infection developed after a dog bite. The infection resolved within 9 days without sequelae after 6 days of ampicillin and clindamycin therapy.


Methionyl-tRNA synthetase from sheep mammary gland. Purification of a fully active monomeric enzyme derived from high-molecular-weight complexes by controlled proteolysis.

Methionyl-tRNA synthetase from sheep lactating mammary gland is found predominantly in the form of high-molecular-weight complexes. Controlled proteolysis of these aggregates generates a low-molecular-weight species of the enzyme with full maintenance of activity as assessed by the rate of aminoacylation of tRNA. The product of proteolysis, which has been purified to homogeneity with a yield of 23%, is a monomeric enzyme of molecular weight 78 000. It has a specific activity of 405 units/mg at 25 degrees C. These findings clearly demonstrate that the aggregated state of methionyl-tRNA synthetase is not a prerequisite for full expression of catalytic activity. Furthermore, the results emphasize the need to provide effective protection against proteolytic damage in studies dealing with the characterization of high-molecular-weight complexes of aminoacyl-tRNA synthetases.


Methionyl-tRNA synthetase from sheep liver. Purification of a fully active monomer derived from high-molecular-weight complexes by trypsin treatment. Evidence for immunological cross-reaction with the corresponding enzyme from sheep mammary gland.

The size distribution of methionyl-tRNA synthetase in extracts from sheep liver is compared to that of lysyl-tRNA, isoleucyl-tRNA, leucyl-tRNA and seryl-tRNA synthetases by gel filtration on Biogel A-5m. Extraction conditions are described which lead to isolation of methionyl-tRNA synthetase exclusively in the form of complexes of molecular weight close to 10(6). Limited trypsin treatment of these aggregates releases a fully active low-molecular-weight form of methionyl-tRNA synthetase which was purified to a specific activity of 674 units/mg at 25 degrees C with a yield of 40%. The homogeneous enzyme appears to be undistinguishable from the corresponding enzyme derived from sheep lactating mammary gland, as judged by acrylamide gel electrophoresis in the presence of sodium dodecyl sulfate and by titration with antibodies raised against the enzyme purified from liver.


Antibody formation and insulin requirements in diabetic children during treatment with purified commercial pork insulins.

40 diabetic children (20 female, 20 male, mean age 8.5 ys, ranging from 0.8 to 14.3 ys), were treated with either of two purified porcine insulin preparations, Komb-Insulin CS (Hoechst) "CS": n = 21, and Semilente MC (Novo) "MC": n = 19, the latter being further purified by ion exchange chromatography. Circulating IgG-antibodies formed in 39 out of 40 subjects during the observation period of more than 2 years. Insulin binding-capacities below 2 mU/ml (mean value after 12 months: 0.45 mU/ml) were found following MC and considerably higher concentrations (mean value after 12 months: 5.16 mU/ml) following CS administration. However, only 52% of the CS-group (n = 11) showed high individual responses (subgroup II), many of which exceeded 5 mU/ml, while the other half (subgroup I) exhibited values as low as those in the MC-subjects. The ratio of girls to boys was high (8 vs 3) in subgroup II, and low (2 vs 8) in subgroup I. Despite great differences in antibody formation, no differences in insulin requirements were noted between the MC- and the CS-groups. Within the CS-group, however, some positive relationship between these parameters was found 6 months and 12 months after the start of therapy, but this did not necessarily apply to individual subjects. Short-term (metabolic control) and long-term effects of therapy (growth) were not different in the MC- and CS-subjects. From these results it may be concluded that antibody formation does not affect insulin requirements or metabolic control during relatively short-term treatment. Provided that antibodies have no direct relation to future vascular changes, both insulin preparations used in this study are equally well-situated for long-term treatment in children and adolescents.


Factor VIII activity and factor VIII related antigen in newborns.

Factor VIII procoagulant activity and factor VIII related antigen were examined in 20 full-term and preterm newborn infants during the first days of life. The control group involved 15 adults volunteers. Factor VIII activity was estimated by a one-stage test and factor VIII related antigen was determined by immunelectrophoresis according to Laurell, using our own rabbit antiserum. The following results were obtained:--Factor VIII activity during the first 3 days of life did not differ from the normal range of the adult controls.--The concentration of factor VIII related antigen in newborns was markedly higher than in adults on the first, and to a lesser extent on the second, day of life.--The antigen concentration decreases on the second and following days of life to adult levels. The cause of this discrepancy cannot be completely explained but possible reasons are discussed.


Is chemical memory transfer due to shock or behavior training?

A memory transfer experiment was performed to determine whether the transfer effect is due to stress associated with the foot shock or to the specific behavioral training. Recipient animals were significantly better in the 'jump-out' testing than either the shock-control recipients or the control recipients, but the shock-control recipients performed significantly better than the control recipients. Foot shock has an effect on the transfer phenomena but is not responsible for the entire effect.


Influence of treatment duration on audiogenic seizure susceptibility during barbiturate withdrawal in rats.

Barbiturate withdrawal seizure susceptibility in rats increased with increasing duration of treatment during a 15-day treatment period in which the animals were given an i.p. dose of sodium barbital every 12 h. This method of producing dependence has clear advantages over previously described methods.


Further characterization of the effects of ultraviolet irradiation of the amphibian egg.

UV-irradiation of the vegetal hemisphere of amphibian eggs leads to developmental abnormalities in neural morphogenesis. The possibility that the egg's transient sensitivity to irradiation could be due to pigmentation changes was examined in albino eggs. The tissue specificity of the effects of irradiation was analyzed by exchanging the ectoderm between irradiated and control embryos.


[The pharmacology of 3,4-dihydroxyphenyl-alpha-hydroxypropionic acid sodium salt].

Sodium salt of 3,4-dihydroxyphenyl-alpha-hydroxypropionic acid inhibits exploratory behavior and induces hypothermia in mice. The compound enhances central effects of L-DOPA, such as: gross excitation in group, fighting behavior, anticataleptogenic action and hyperthermic effect. The L-DOPA central effects are mitigated by pyridoxal phosphate. The 3,4-dihydroxyphenyl-alpha-hydroxypropionic acid sodium salt abolishes the protective action of pyridoxal phosphate. In vitro this salt inhibits DOPA-decarboxylase. It is suggested that the compound under discussion is a competitive antagonist of L-DOPA, when the latter is involved in reactions with pyridoxal phosphate.


Effect of chlorpromazine on the in vitro immune response.

The effect of chlorpromazine at concentrations from 10(-7) to 5 X 10(-5)M on the in vitro plaque-forming cell responses to sheep erythrocytes was tested. If CPZ was present in cultures during the whole incubation period, concentrations of CPZ more than 5 X 10(-6)M inhibited the PFC responses, whereas concentrations between 5 X 10(-7) and 5 X 10(-6)M had a slightly enhancing effect. If a defined concentration of CPZ (1.3 or 1.5 X 10(-5)M) was added to cultures for a brief period of time (12 or 24 h) at various intervals after the initiation of culture and antigenic stimulation, PFC responses were inhibited most effectively during the first brief periods of incubation. The later the drug was added, the more the extent of inhibition decreased. On the other hand, CPZ had no effect or a slightly enhancing effect when added in the very late period (72--95 h) of incubation. These findings suggest that the observed immunosuppression could be attributed mainly to the impairment of early cellular events, presumably cell interaction events, preceding the process of DNA synthesis and cell proliferation of antibody-forming cells and their precursors.


[The effect of non-steroid antiphlogistic drugs on the number of antibody-forming cells during the primary immune response].

In mice immunized with sheep erythrocytes introduction of acetylsalycilic acid (250 mg/kg), sodium salicylate (100 mg/kg) and mephenamic acid (100 mg/kg) exerted an inhibitory effect on the synthesis of antibody-forming cells and the titre of hemagglutinines in immunized animals. The acetylsalicylic acid and sodium salicylate are more effective on the 3--5th post-immunization day; the maximal effect of the mephenamic acid becomes apparent on the 7th day and its action is more lasting.


Studies on the retest reaction in contact sensitivity to DNCB.

DNCB-sensitized guinea pigs demonstrated an accelerated reactivity on retest of DNCB at the site of prior contact reaction, though presenting normal contact sensitivity at the virgin site. The retest reaction reached maximal at 9 h and waned at 24 h after antigenic challenge. Massive accumulation of eosinophils in either the epidermis or dermis was its distinguishing histologic feature. The reaction was induced at the site of delayed skin reaction to DNP-GPE in the animals sensitized with DNCB or DNP-GPE. A retest reaction in delayed sensitivity to DNP-GPE was also elicited at the site of contact reaction to DNCB in the animals. The significance of these findings is discussed.


Perinatal malnutrition and early handling: interactive effects on the development of the pituitary-adrenal system.

The preweaning development of the pituitary-adrenal system was investigated in perinatally malnourished rodents. Malnourished pups that were routinely handled displayed an earlier plasma corticosterone stress response to electric shock and an accelerated development of feedback inhibition of the stress response by exogenously elevated steroid levels. Although malnourished pups displayed elevated basal corticosterone levels compared to well-fed controls, the post-stress levels were also higher in these malnourished young. Thus, the corticosterone response to stress was not impaired in malnourished pups. The influence of the early handling of malnourished young on pituitary-adrenal activity was also studied. Whereas the accelerated development of the stress response and the elevated post-shock levels were not the result of early handling, the elevated basal corticosterone levels displayed by handled malnourished pups were not present in nonhandled malnourished pups. Thus, the early non-nutritional environment interacts with malnutrition to alter the development of the pituitary-adrenal system.


A neuropharmacological analysis of central nervous system catecholamine systems in development protein malnutrition.

Four experiments were performed to evaluate the effects of developmental protein deprivation on the behavioral response of adult rats to treatments known to affect central nervous system catecholamine systems. Results showed no group differences between protein malnourished and control animals in locomotor responsiveness to d- or l-amphetamine, recovery from behavioral asymmetry produced by a unilateral lesion of the substantia nigra, or in the development of response patterns indicative of denervation supersensitivity. However, a dose-dependent diminution in the ability of apomorphine to produce stereotyped behavior was noted in the malnourished group, suggesting that a class of brain dopamine receptors may be impaired or may have undergone homeostatic modification as a result of the undernutrition procedure.


[Primary dysmenorrhoea and hyperprolactinemia (author's transl)].

Between 5 and 10% of all young girls and women suffer of severe primary dysmenorrhoea which interfers monthly with their activities. In this type of primary dysmenorrhoea we found hyperprolactinemia in 13 of 15 women who were examined. All 15 patients had a pituitary tumour ruled out and were treated with Bromo-ergocryptine (parlodel). Eight patients became free of symptoms and five were well improved. The possible correlation of hyperprolactinemia to premenstrual tension and severe primary dysmenorrhoea is discussed.


[Genetic aspects of the effect of Vi-antigen deposition in the human body. I. Hereditary cause of the effect of Vi-antigen deposition on erythrocytes].

Investigation of the minimal and the optimal capasity of the Vi-antigen sorption in erythrocytes was carried out in partners of 11 pairs of monozygotic and 8 pairs of dizygotic twins. It is shown that the capacity of erythrocytes to sorb Vi-antigen in minimal concentrations (Smin) is under relatively rigid genetic control (G = 0.84), while the capacity of its sorption in optimal doses (Sopt) is to a much greater extent subjected to paratypic effects (G = 0.40). It is assumed that Smin is controlled by the system of the "main" gene, while Sopt is determined multifactorially, which affords the possibilities of creating adequate conditions for the vaccination of a concrete individual.


Timing of gestation in rats by fetal and maternal weights.

Weights of embryonic and fetal rats increase with gestational age from Day 13 to 20 in a strictly logarithmic manner and then slow. Data for other rat strains published as long ago as 1915 show substantially the same growth curves and the same weights as the inbred Kx strain studied. During this late gestational period fetal weights doubled every 1.22 days. An earlier period of somewhat faster growth can be inferred. These growth rates are virtually the same as the maximum rates at which rat neoplasms can grow. A regression equation was constructed from which the ages of rat embryos can be predicted from their weights within confidence limits of +/- 0.24 days. A similar logarithmic increase of added weight in the pregnant dam was recognized by which pregnancies could be conveniently diagnosed as early as Day 7. These two procedures facilitate the routine production of embryos of known age for experimental purposes.


[Influence of estrogen on the delayed action of prostaglandin F2alpha on the rat uterine contraction (author's transl)].

Responsiveness of the rat uterus on the day of estrus to PGE2, PGF2alpha and oxytocin was increased by administration of PGF2alpha i.p. 24 hours prior to the experiments, but that of the uterus on the day of diestrus was not altered by the same treatment. The delayed action of PGF2alpha was not observed in the uterus of ovariectomized rats, but was observed in the uterus of estrogen treated rats. The delayed action of PGF2alpha was not evident in either the vas deferens or the intestine of the rat.


Effect of malonate and maleate on growth and brefeldin A formation in Curvularia lunata.

Malonate exerted a stronger inhibitory effect on brefeldin A production than on mycelia growth in cultures of Curvularia lunata especially at inhibitory levels of 100 to 200 mM. The extent of 200 mM malonate inhibition of growth and brefeldin A production was greater in cultures treated with malonate prior to inoculation than those treated following 5 days after inoculation. Maleate at levels of 40 to 220 mM activated brefeldin A formation in cultures though exerting variable effects on mycelia growth.


Diurnal changes in serum metabolites and hormones in lean and obese Zucker rats.

A study of diurnal patterns of serum hormone levels was made in genetically lean and obese Zucker rats. Serum glucose and insulin were elevated in the obese through the 24-hour period; immunoreactive insulin being 12 times higher in the obese rat. A similar pattern of serum hormone levels in lean and obese rats was found for triiodothyronine, thyroid stimulating hormone and prolactin. At peak levels, prolactin was 2.5 times higher in lean than obese rats (P is less than .01). Thyroxine levels tended to be lower in the obese rats throughout the sampling periods when compared to lean rats. Growth hormone levels were severely depressed in the obese rat with the largest differences between lean and obese rats being observed during the light periods of the cycle. These differences in growth hormone secretion may be responsible for the impaired nitrogen retention in the obese rat. Serum levels of corticosterone were highest at the beginning of the dark cycle and decreased drastically thereafter in the lean rats. The obese rat did not exhibit a distinct pattern of corticosterone secretion and tended to be elevated throughout the periods tested.


[Preparative isolation of di-, tri- and tetrapyrimidine nucleotides from depurinated herring sperm DNA (author's transl)].

The pyrimidine nucleotides p(dT)3, p(dT)2p, pdCp, p(dC)2p and p(dC)3 and the mixtures of sequence isomers p(dC, dT), p(dC, dT)p, p(dT2, dU), (dT2, dU)p, p(dC, dT2), p(dC, dT2)p, p(dC, dT3), p(dC, dT), p(dC2, dT)p, and p(dC2, dT2) are isolated on a preparative scale from depurinated hydrolysates of herring sperm DNA by the following procedure. The DNA hydrolysate is first separated into a low and a high molecular weight mixture of pyrimidine nucleotides by column chromatography on DEAE-cellulose. The nucleoside bisphosphates pdCp and pdTp, the dinucleotides p(dC)2 and p(dT)2 and the sequence isomers p(dC, dT) are largely separated out of the mixture of the low molecular weight pyrimidine nucleotides. The remaining mixture is rechromatographed at pH 3.5 on QAE-Sephadex. This separates the pyrimidine nucleotides containing a majority of cytidylic acid units from those containing more thymidylic acid units, which are then fractionated at pH 7.5 according to the number of bases in the chain. The pyrimidine nucleotides and mixtures of sequence isomers separated to 83-99% purity by column chromatography are further separated by paper chromatography and are obtained in chromatographically pure form after this step. The structures of the isolated DNA fragments are determined from chromatographic data, absorption and enzymatic degradation.


Ultrastructural findings supporting the angioblastic nature of the so-called adamantinoma of the tibia.

Electron microscopy of a case of adamantinoma of the tibia shows features which support a mesenchymal angioblastic origin rather than epithelial. Comparison is made between tissue from this tumour and a squamous cell carcinoma of the femur arising in chronic osteomyelitis. In adamantinoma no desmosomes are found between tumour cells and their cytoplasmic ultrastructure shows features of mesenchymal cell type including evidence of pinocytic activity and bundles of filaments resembling hyperplastic endothelial cells. The stroma shows features similar to fibrous dysplasia of bone with fibroblasts, histiocytes and fibrolipoblastic lipid-laden mesenchymal cells. It is condluded that adamantinoma of the tibia should be considered to be an angioblastic tumour of bone.


Hospitals in Wonderland.

The hospital industry faces a choice. It can follow a path of involvement with HEW and HMOs, with the prospect of eventually being dominated by them, or it can work toward a position of neutrality, attempting energetically to influence Congress on behalf of the legitimate concerns of both hospitals and government.


Furrows and dermal ridges of the hand in patients with alcohol embryopathy.

Palmar creases and dermal ridge patterns of 34 patients with alcohol embryopathy are compared with 470 healthy individuals. In alcohol embryopathy several typical deviations were noted. Palmar Creases. The interdigital part of the distal palmar crease is generally sharply bent, the proximal transverse crease is hypoplastic or missing, the thenar crease is commonly well marked. Simian creases and bridged palmar creases are more common in patients with alcohol embryopathy than in healthy individuals. Ridge Patterns of the Palm. The main line D coming from triradius d in patients with alcohol embryopathy mostly shows a low type of ending in the fourth interdigital area; in this area loops are twice as common as in healthy individuals. Patterns of the Fingertips. No deviations were noted in the distribution of whorls and loops, but virtually no arches were observed in patients with alcohol embryopathy. These anomalies suggest embryonic damage in the twelfth week of gestation.


Time and cell systems as variables in fusion experiments with polyethylene glycol.

Cell hybridization was done between a monolayer of B14-150 Chinese hamster cells and a suspension of either mouse leukemia cells or normal human lymphocytes. Cell contact was obtained by centrifugation of the suspension cells onto the monolayer cells in a culture plate. Cell fusion was done by means of polyethylene glycol (PEG). The optimum time for PEG exposure as well as the yield of hybrid cells differed markedly with the different combinations.


Balanced reciprocal whole-arm translocation t(1;19) in three generations.

Seven carriers of a balanced reciprocal whole-arm translocation of chromosomes 1 and 19 were found in 19 members investigated in three generations of one family. The apparent lack of a carrier of an unbalanced form of the translocation and of a history of miscarriages are discussed.


47,X,i(Xq),Y karyotype in Klinefelter's syndrome.

This is a case report of 47,X,i(Xq),Y in a 24-year-old infertile male with Klinefelter's syndrome. C staining indicated that this isochromosome X had a single small centromere. BUdR incorporation revealed the isochromosome X to be late replicating.


No elevation of exchange type aberrations in lymphocytes of children with alcohol embryopathy.

The lymphocyte chromosomes from 23 children with the 'fetal alcohol syndrome' or 'alcohol embryopathy' (AE) were analyzed with respect to exchange type aberrations. These aberrations were not more frequent in AE than in controls. The possibility that AE results from unspecific suppression of RNA and consequently of protein synthesis in the developing embryo is discussed.


Establishment and characterization of a cell line from the American opossum (Didelphys virginiana).

A permanent tissue-cultured cell line (designated OK) has been established from kidney tissue of an adult American opossum. The OK line has been characterized with respect to morphology, chromosome constitution, tissue-culture requirements, and attainable mitotic arrest. The cells are epithelial-like with a stable nondiploid chromosomal modal number of 23. Cells grown in Eagle's minimal essential medium with 10% fetal bovine serum have a mean doubling time of 18 hr. The cell line OK is potentially useful for the isolation and purification of the mammalian X chromosome because of the size differential between the smaller X's and the larger autosomes.


The effect of incubation temperature on the maintenance of rat palatal mucosa in organ culture.

The effect of incubation temperature on the behavior of neonatal rat palatal mucosa maintained in a chemically defined medium in organ culture for periods up to 7 days was investigated. Explant survival was optimal at 37 degrees C with increasing mortality at temperatures of 34 degrees C and 30 degrees C. There was a transient increase in the epithelial mitotic activity at all temperatures, but at all time intervals mitotic activity was greatest at 37 degrees C. While the mitotic activity at 37 degrees C after 5 hr in vitro was comparable with previously described in vivo values, it was subsequently increased, only returning to values approximating those at the start of the experiment at 6 days. At 30 degrees and 34 degrees C the epithelial mitotic activity increased more slowly than at 37 degrees C; then it followed a similar pattern with time and after 5 days in vitro had fallen to values approximating initial values. At the cut edges of the explants, the rate of epithelial migration and subsequent keratinization increased with increasing temperature. It is suggested that survival of neonatal rat palatal mucosa is optimal in this organ culture system when maintained at 37 degrees C.


Studies on the transformation of rat embryo cells of low passage by carcinogenic fluorenylhydroxamic acids and their acetate esters.

Rat embryo cells of low passage subjected to a single treatment with certain carcinogenic fluorenylhydroxamic acids and their respective acetate esters showed signs of transformation in vitro, such as changes in phenotype, growth in soft agar and agglutination with concanavalin A. In addition, certain changes in karyotype and loss of diploidy were observed. There was no evidence, either by electron microscopy or by assay of RNA-dependent DNA polymerase, for the presence of virus. None of these cell lines produced tumors after inoculation into the isologous host. The results of these study lead us to suggest that malignant transformation is a multistep process and that certain criteria of transformation of rat embryo cells are associated with the initial stage(s) in which the cells are transformed without being tumorigenic. The ultimate test for malignant transformation of rat embryo cells remains the production of tumors in a susceptible host after inoculation of treated cells.


Temperature and species differences in susceptibility of kidney cell cultures to mercury toxicity.

The effect of temperature on inorganic mercury toxicity was investigated using kidney tissue culture systems. The relative susceptibility of rabbit (homeothermic) kidney to mercury intoxication was compared to that of Coho salmon (poikilothermic) kidney to mercury intoxication was compared to that of Coho salmon (poikilothermic) kidney over temperature ranges consistent with the habitat of each of the two species. It was demonstrated that susceptibility to mercury toxicity is species dependent; that is, the rabbit kidney cells tolerated higher mercury concentrations in the medium than did the fish-derived cells. Within a given species, susceptibility to mercury toxicity was temperature dependent. Decreasing the temperature increased the toxicity of mercury to cultures of rabbit kidney cells, whereas decreasing temperatures decreased the effect of mercury toxicity on the salmon kidney cells. As a consequence, fish taken from arctic waters are liable to be more toxic when introduced into mammalian food chains. Albumin was shown to act as a protective agent in vitro against inorganic mercury toxicity.


Primary monolayer cultures of postnatal rat liver cells with extended differentiated functions.

Monolayers of liver cells cultured from postnatal rats were grown in two types of media. One set of cultures was grown in selective medium which contained ornithine but was deficient in arginine; the other set was grown in nonselective medium which contained arginine but no ornithine. The cultures that were grown in the nonselective medium contained primarily a mixture of two cell types found in the liver: parenchymal hepatocytes and fibroblast-like cells. The fibroblast cells tended to overgrow the hepatocytes after several days in culture. In contrast, fibroblast overgrowth was inhibited in cultures grown in the selective, arginine-deficient medium, thereby resulting in relatively pure cultures of functional parenchymal hepatocytes. Comparative studies of sulfobromophthalein (BSP) uptake showed that the cultures grown in selective medium continued to be active much longer than the cultures grown in the nonselective medium. Pyruvate kinase assays revealed that the cultures grown in selective medium contained primarily the L-isoenzyme type which is characteristic of parenchymal hepatocytes. Cultures grown in nonselective medium contained a mixture of L- and M-isoenzymes which is indicative of nonparenchymal liver cells. The reported results indicate that selective, arginine-deficient medium permits primarily the growth of parenchymal hepatocytes found in neonatal rat liver.


Survey of ATCC stocks of human cell lines for HeLa contamination.

Seed stocks of human cell lines deposited at the American Type Culture Collection (ATCC) have been examined for cross-contamination with HeLa cells using Giemsabanded marker chromosomes. Sixteen additional cell lines investigated have been found to exhibit marker chromosomes typical of HeLa cells. Quinacrine fluorescence studies further revealed the absence of Y chromosome in these lines. These observations indicate that the lines are HeLa derivatives.


Isolation and characterization of epithelial cells from bovine pancreatic duct.

Epithelial cells derived from bovine pancreatic duct have been grown continuously in culture for 30 weeks (approximately 90 doublings of the cell population). The cells were grown in Eagle's minimal essential medium supplemented with 10% heat-inactivated fetal bovine serum, 2 mM glutamine, 0.1 mM nonessential amino acids, and antibiotics. In confluent cultures, the cells are multilayered and form circular structures. When tested at various passages, the cells neither formed colonies in soft agar nor produced tumors after inoculation into athymic, nude mice. Hydrocortisone (1 and 5 microgram per ml) and insulin (1,5 and 10 microgram per ml) had no effect on the growth of the cells. beta-Retinyl acetate inhibited growth rate and cell yield at a concentration of 5 microgram per ml but was not growth-inhibitory at lower concentrations. By electron microscopy the cells have numerous mitochondria, Golgi and microvilli. Mucous droplets were observed in a small proportion of the cells. Desmosome-like structures and occluding junctions were observed more frequently between cells that had been transferred as aggregates than between cells transferred as single cells. Cytochemical studies indicated that some cells produce PAS positive granules that were not removed after treatment of the cultures with diastase. Eleven cell clones were isolated from the mass culture. The growth rates of the clones are different as well as the period of time in which the clones can be propagated in vitro.


[Thrombocyte aggregation after vaginal hysterectomy with combined prevention of thromboembolism with macrodex 6% and sintrom].

The ADP- and collagen-induced thrombocyte aggregation has been investigated and statistically evaluated in 25 women. These patients had undergone vaginal hysterectomy and colporrhaphy due to descensus vaginae et uteri and received 500 ml Macrodex 6% during the operation and Sintrom from the 1st post-operative day as prophylactic treatment for early and late thromboembolism. After administration of dextran 60, the ability of the thrombocytes to aggregate was reduced by half, 2 hours post-operatively, independent of the concentration and type of agent added in the aggregation test, and remained at this level during the 1st post-operative day. The original values were once again attained on the 4th post-operative day. The speed and force of deaggregation showed reverse behaviour. There was no influence apparent due to age of the patient or phase of the cycle (proliferation, secretion and post-menopausal). By carrying out this combined prophylactic treatment for thromboembolism, satisfactory protection is provided in the dangerous phases during the operation and post-operatively.


Cultures of Chlamydia trachomatis in mouse peritoneal macrophages: factors affecting organism growth.

Growth of Chlamydia trachomatis B/TW-5/OT and L(2)/434/Bu strains in cultures of thioglycolate-activated mouse peritoneal macrophages was studied. Both strains grew to a limited extent in the macrophages, but lymphogranuloma venereum (LGV) grew better than trachoma. Growth was enhanced by centrifugation of the inoculum onto the macrophage cell layer and inhibited by pretreatment of macrophages for 2 h with 100 mug of concanavalin A per ml. No significant effect was observed by pretreatment of macrophages with diethylaminoethyl-dextran (30 mug/ml, 30 min), cytochalasin B (20 mug/ml, 1 h), and cyclophosphamide (200 mug/ml, overnight) or by treatment with hydrocortisone (1 and 10 mug/ml, overnight before inoculation and during a 2-day incubation after inoculation). Resistance to intracellular growth of the two organisms was not increased in macrophages obtained from mice immunized with the organisms compared with macrophages from nonimmunized mice unless they were pretreated in vitro with penicillin (100 U/ml, overnight). The yields of LGV organisms from the penicillin-pretreated macrophages of LGV-immunized mice were 100-fold less than from the pretreated macrophages of nonimmunized control mice. At the same time, the yields of LGV organisms from penicillin-pretreated macrophages of mice immunized with trachoma, gonococcus, and HeLa cells were not different from those obtained in pretreated macrophages of nonimmunized control mice.


Correlates of alcohol arrests in a rural state.

Arrest and conviction rates for public intoxication, driving while intoxicated, and liquor law violations were correlated with population, population change, liquor sales, number of law officers, Indian population, and overall crime rate for 23 counties in Wyoming. It was concluded that increased liquor sales, but not arrest rates, are associated with population increase. Alcohol arrests are related positively to population magnitude and overall crime rate, but not to number of law officers. Counties with larger Indian populations report higher arrest rates for public intoxication and liquor law violations, but conviction rates for alcohol arrests in these counties are not different.


Studies on trypsin inhibitors. Part VIII. Synthesis of the protected octatriacontapeptide corresponding to the sequence 15-52 of porcine pancreatic secretory trypsin inhibitor II (Kazal).

The synthesis by fragment condensation of protected peptides corresponding to the amino acid sequences 15-35, 25-52 and 15-52 of porcine pancreatic secretory trypsin inhibitor II (Kazal type) is described. The Rudinger modification of the azide procedure was used in the fragment coupling steps. The tert-butyloxycarbonylheptapeptide hydrazide (sequence 22-28) was reacted with the heptapeptide methyl ester free base (sequence 29-35) and the resulting tert-butyloxycarbonyltetradecapeptide methyl ester after selective deprotection, coupled with the benzyloxycarbonylheptapeptide hydrazide (sequence 15-21) to give the protected peptide methyl ester corresponding to the 15-35 sequence which was then converted to the corresponding hydrazide. The synthesis of the 25-52 sequence was achieved by assembling the protected peptide hydrazide corresponding to the amino acid residues 25-35, with the C-terminal heptadecapeptide 36-52. The resulting protected octaeicosapeptide (sequence 25-52) was selectively deblocked with trifluoroacetic acid and acylated with the benzyloxycarbonyldecapeptide hydrazide 15-24 to give the desired octatriacontapeptide corresponding to sequence 15-52 of the inhibitor. An attempt to prepare the 15-52 sequence through the condensation of fragments corresponding to 15-35 and 36-52 sequences was unsuccessful. The identity and purity of the synthetized peptide derivatives wre established by elemental analysis (in some cases), amino acid analysis, optical rotation, and thin-layer chromatography in two solvent systems. The final products were also evaluated, after partial deprotection with anhydrous hydrogen fluoride or aqueous 90% trifluoroacetic acid, by paper electrophoresis at different pH values.


The presence of lysosomal hyaluronidase in the rabbit iris.

Lysosomal hyaluronidase in the rabbit iris was studied by means of a sensitive assay method based on carbocyanine dye binding. The enzyme activity in the lysosomal extract was proportional to both the enzyme concentration and the incubation time. When the enzyme was heated, the enzyme activity was lost completely. When the lysosomal extract was used as an enzyme source, the enzyme activity was protected by pepstatin. The enzyme had a pH optimum of 3.8 and no activity above pH 5.0.


Association between thyrotoxicosis and thrombocytopenia. A case report and review of the literature.

In a young woman who presented with hyperthyroidism and autoimmune thrombocytopenic purpura, the platelet count returned to normal following successful treatment of the hyperthyroidism. Thromboagglutinins were present, the titer declining as the patient became euthyroid. A survey of the literature revealed 48 reports of hyperthyroidism and thrombocytopenia, these disorders coexisting in 37 patients and there being no apparent cause for the lowered platelet counts in 28 of them. The clinical features and response to treatment in the latter group and in our patient are reviewed, comprising a series of 29. Of 22 patients whose hyperthyroidism was adequately treated, platelet counts returned to normal in 18 (82%). In three patients the purpura remitted despite persistent thrombocytopenia. It is estimated that in 7% of patients with autoimmune thrombocytopenic purpura, thrombocytopenia responds to treatment of an underlying thyrotoxicosis and in many of these patients the thrombocytopenia will prove resistant to other forms of therapy.


Prepubertal XX male with profound physical and mental deficiency, retinitis pigmentosa and multiple congenital anomalies.

A unique case of a prepubertal XX male with profound mental and physical retardation, retinitis pigmentosa, ambiguous genitalia and multiple congenital anomalies is reported. His clinical, genetic, dermatoglyphic and histological findings are presented. This case could represent a new multiple congenital malformation syndrome. Theories on XX male aetiology are briefly discussed.


Leaching of glycosaminoglycan from cow uterus and vagina into fixative solution.

Samples from the genital tract of two cows in metoestrus and two in dioestrus were fixed with formalin-cetyltrimethylammonium bromide and the amount of glycosaminoglycan leached into this histological fixative was estimated using an Alcian Blue method. Uronic acid content was estimated by a carbazole method. Total glycosaminoglycan was estimated after digestion with papain. No more than 10% of the glycosaminoglycan was soluble in the fixative and in comsequence 90% is potentially stainable by  Alcian Blue in tissue sections. Total glycosaminoglycan in uterus and vagina is higher in metoestrus than in dioestrus. Estimation of uronic acid in the leached material confirmed the results and indicated that little of it consisted of mucin or keratan sulphate.


Some practical considerations in the assessment and treatment of adolescent girls with separation problems.

Five cases are presented of adolescent girls with severe problems in leaving home. Though the psychodynamics were similar (preoedipal conflicts in the separation-individuation phase), the psychopathology and underlying causative factors were different in each case. Factors to be considered in the initial diagnostic assessment are outlined. These include constitutional and congenital factors, the developmental history, maternal emotional illness, and gene-environment interactions. Various treatment modalities are described and illustrated. In four of the cases specific parameters were necessary, particularly behavioral technics and conjoint family therapy. The manner in which the analyst may serve, in cases amenable to psychoanalytic treatment, as transference object plus real object is discussed and illustrated.


Effect of calcium on ion transport and electrical properties of tracheal epithelium.

Active transport of Cl- toward the tracheal lumen and Na+ away from the lumen creates an electrical potential difference across dog tracheal epithelium. This study examined in vitro the effect of varying calcium concentration in the bathing media on the ion transport and electrical properties of dog tracheal epithelium. In six pairs of epithelia, changing calcium concentration from 1.9 to 0 mM resulted in a significant decrease in electrical resistance, from 318 +/- 36 to 214 +/- 24 omega.cm2. Short-circuit current and net Cl- and Na+ fluxes measured under short-circuit conditions were not changed significantly. Changing calcium concentration from 1.9 to 10 mM resulted in no significant change from control in the electrical properties nor in net Cl- and Na+ fluxes (short-circuit conditions). Histamine (10(-4) M) produced a significantly smaller increase in short-circuit current in 0 than in 1.9 mM Ca2+ (+5 +/- 2 vs. +12 +/- 2 microamperemeter/cm2). However, electrical changes were not significantly different in 1 or 10 mM Ca2+. These results indicate that calcium lack increased permeability of tracheal epithelium and that the increase in short-circuit current due to histamine depended in part on calcium.


High Dipetalonema reconditum microfilarial counts in two dogs.

Dipetalonema reconditum microfilarial counts of 509 and 932/ml of blood, respectively, were found in 2 dogs thought to have been infected with Dirofilaria immitis. The high counts were attributed to a state of immunosuppression. It was concluded that microfilarial width is a more reliable criterion that microfilarial numbers for the differentiation of D immitis and D reconditum infections. For reliable width measurements, it is necessary to use the modified Knott or polycarbonate filter techniques, inasmuch as the cellulose filter technique gives misleading results.


Central nervous system depression associated with glucocorticoid ingestion in a dog.

Signs of a central nervous system disorder were observed following 2 instances of accidental ingestion of glucocorticoid in a young female Doberman Pinscher. The signs included transient aggressive and paranoid behavior, amaurosis, disorientation, ataxia with circling backward, and depression. Vomiting, weight loss, and abnormal drinking behavior persisted for several weeks following recovery from the acute illness.


Effects of endometrial biopsy, uterine culture, and cervical dilatation on the equine estrous cycle.

In 10 mares, lysis of the corpus luteum was induced by endometrial biopsy and culture on day 4 after estrus, as evidenced by a sharp decline in serum progesterone concentration and a shortened interestrous interval. Estrus following the manipulations was prolonged. Endometrial biopsy and culture on days 1 and 3 after estrus or manually dilating the cervix on day 4 after estrus also decreased the serum progesterone concentration (within 24--48 hr after manipulation), shortened diestrus, and prolonged the subsequent estrus. In a 2nd experiment, endometrial biopsy on day 4 after estrus shortened the estrous cycle and diestrous period and prolonged the subsequent estrus. It was postulated that these manipulations stimulated corpus luteum regression, followed by estrus and ovulation in the mare.


Differential development of autistic symptoms in a pair of fraternal twins.

The following case study reports the differential development of autistic symptoms over a 9-year period in a pair of fraternal twins. Neither of the twins developed normally. Evaluations revealed that although the older (male) twin showed some early signs of pathology and suffered from frequent illnesses, he ultimately has developed more normally than his sister, who likely sustained brain damage as a result of perinatal anoxia. Significant improvements has occurred in both cases. Diagnostic, etiological, and treatment implications are discussed.


Effect of sodium chloride on changing the rate-limiting step in the human placental choline acetyltransferase reaction.

The kinetic constants, Km and Vmax, for the choline acetyltransferase reaction were determined for choline and eight choline analogs under conditions of high (0.3 M) and low (approximately 0.01 M) sodium chloride. At high sodium chloride, the maximal velocities of the different substrates varied over 27-fold, while at low sodium chloride, less than a 5-fold variation was observed. Dead-end inhibition studies using acetylaminocholine as the inhibitor showed that under conditions of high sodium chloride, inhibition changes from noncompetitive to competitive as the reactivity of the substrate decreases. Under conditions of low sodium chloride, acetylaminocholine inhibition is nonlinear and noncompetitive with respect to all substrates tested. These results suggest that increased ionic strength increases the rate of coenzyme A dissociation from the enzyme. The rate-determining step of the reaction can be ternary complex interconversion, coenzyme A release, or both, depending on the ionic strength and the substrate employed.


Purification and characterization of the sex steroid-binding protein of rabbit serum. Comparison with the human protein.

The sex steroid-binding protein (rSBP) of immature rabbit serum was purified to homogeneity by the sequential use of DEAE-cellulose chromatography, affinity chromatography on 5alpha-dihydrotestosterone-17 beta-succinyl-diaminoethyl-(1,4-butanediol diglycidyl ether)-agarose, agarose (Bio-Gel-A-0.5m) gel filtration, and preparative polyacrylamide gel electrophoresis. The cumulative yield is 13%. Homogeneity of rSBP was shown by the equilibrium sedimentation ultracentrifugation in 6 M guanidine HCl containing 0.1 M mercaptoethanol which yields an average molecular weight of 36,475 +/- 865. Analytical gel electrophoresis in the presence of sodium dodecyl sulfate and gel filtration on agarose yield a molecular weight of 57,000 and 120,000, respectively. The variation is due to a 30% carbohydrate content. The amino acid composition is reported. Comparison of the rabbit and human SBP indicate that they are different in both their molecular and functional properties.


Topical hemostatic agents for bone bleeding in humans. A quantitative comparison of gelatin paste, gelatin sponge plus bovine thrombin, and microfibrillar collagen.

The three topical hemostatic agents--gelatin paste, microfibrillar collagen, and gelatin sponge soaked in thrombin--individually were effective in reducing bleeding from cancellous bone, as tested on the femoral surface of trochanteric osteotomies during total hip replacement. During a three-minute interval, the spontaneous reduction in bleeding in eight control hips to which no agent was applied was 11 per cent. Gelatin paste gave a reduction of 85 per cent; gelatin sponge soaked in thrombin, a reduction of 75 per cent; and microfibrillar collagen, a reduction of 47 per cent. None of these agents interfered with healing of the trochanteric osteotomy.


Secretory kinetics in the follicular cells of silkmoths during eggshell formation.

Procedures for quantitative autoradiography were used for studying the process of secretion of eggshell (chorion) proteins in the follicular epithelium of silkmoths. The method was based on photometric measurements of the reflectance of vertically illuminated autoradiographic silver grains. Results were analyzed and plotted by computer. Secretory kinetics were also determined by analysis of labeled proteins in physically separated epithelium and chorion. Rapid accumulation of radioactivity into "clumps" visualized by light microscope autoradiography and evidence from preliminary electron microscope autoradiography indicate that, within 2 min from the time of synthesis, labeled chorion proteins move to Golgi regions scattered throughout the cytoplasm. The proteins begin to accumulate in the apical area 10-20 min later and to be discharged from the cell. The time for half-secretion is 20-25 min, and discharge is essentially complete 30-50 min after labeling. At the developmental stages examined, the kinetics of secretion appear to be similar for all proteins. Within the chorion the proteins rapidly assume a characteristic distribution, which varies for different developmental stages. Two relatively slow steps have been identified in secretion, associated with residence in Golgi regions and in the cell apex, respectively. By contrast, translocation of proteins across the cell and deposition of discharged proteins in the chorion are rapid steps.


Mechanism for the selection of nuclear polypeptides in Xenopus oocytes.

The function of the nuclear envelope in regulating the cellular distribution of proteins was studied by experimentally altering nuclear permeability and determing the effect of the procedure on the incorporation of exogenous and endogenous polypeptides into the nucleoplasm. Using fine glass needles, nuclear envelopes were disrupted by puncturing oocytes in that region of the animal pole occupied by the germinal vesicle. This resulted in a highly significant increase in the nuclear uptake of cytoplasmically injected [125I]-bovine serum albumin ([125I]BSA), deomonstrating that the envelopes had lost their capacity to act as effective barriers to the diffusion of macromolecules. Endogenous proteins were labeled by incubating oocytes in L-[3H]lecuine. After appropriate intervals, nuclei were isolated from punctured and control cells and analyzed for tritiated polypeptides. Both total precipitable counts and the proportion of label in different size classes of polypeptides were compared. The results showed that puncturing the oocytes had no apparent quantitative or qualitative effects on the uptake of endogenous polypeptides by the nuclei. It can be concluded that the accumulation of specific nuclear proteins is not controlled by the envelope but rather by selective binding within the nucleoplasm.


Thin (actin) and thick (myosinlike) filaments in cone contraction in the teleost retina.

The long slender retinal cones of fishes shorten in the light and elongate in the dark. Light-induced cone shortening provides a useful model for stuying nonmuscle contraction because it is linear, slow, and repetitive. Cone cells contain both thin (actin) and thick (myosinlike) filaments oriented parallel to the axis of contraction. This study examines the polarities of the cone's thin filaments and the changes in filament distribution which accompany light-induced contraction, in an attempt to elucidate the structural basis for the cone's contractile process. The proximal half of the cone is fixed to its cellular neighbors in the outer nuclear layer while the distal half is free. Thus, all shortening takes place in a necklike region (the myoid) in the distal half of the cone which extends into the space between the neural retina and the pigmented retinal epithelium. Thin filaments are found throughout the length of the cone, whereas thick filaments occur predominantly in the proximal (axon) regions of both light- and dark-adapted cones. Thus, thick filaments are primarily localized outside the region where shortening takes place. Observations from myosin subfragment-1 binding studies suggest that the cone's thin filaments are organized into two opposing sets. In the distal half of the cone (including the myoid), virtually all filaments have proximally directed arrowheads. In the more proximal regions of the axon, many thin filaments have opposite polarity, their arrowheads being distally directed. Near the synaptic proximal end of the light-adapted (contracted) cone, filaments of opposite polarities occur in approximately equal numbers. Thus, in the cone axon there appear to be two overlapping sets of actin filaments whose opposite polarities correspond to the two actin halves of a muscle sarcomere. In elongated, dark-adapted cones, thick filaments are localized throughout the axon region of the cone. In light, thick filaments accumulate towards the proximal end of the cone. These observations are consistent with a "sliding hypothesis" for cone contraction, in which thick myosinlike filaments produce sliding interdigitation of the two sets of oppositely directed actin filaments in the proximal axon region. Thus, the myoid thin filaments would be essentially reeled into the axon region to produce shortening. The mechanism of re-elongation depends on microtubules, as discussed in the companion paper.


Actin content and organization in normal and transformed cells in culture.

The amount of actin and total protein per cell in normal rat kidney (NRK) cells in culture is initially high in very low density cultures, but rapidly decreases as the cells come into contact in higher density cultures. In a viral transformant of NRK (442), the level of actin and total protein does not change significantly from low to high density cultures. NRK cells, which are flattened against the substrate, have prominent bundles of actinlike microfilaments in the basal cytoplasm adjacent to the substrate. 442 cells, which adhere poorly and are more spherical in shape, lack well-organized basal microfilament bundles, but may display microfilament bundles in cytoplasmic processes extending from the cell body. The percentage of insoluble actin is less than 20% in both cell lines, and 442 cells consistently contain smaller amounts than NRK cells.


Laser microbeam study of a rotary motor in termite flagellates. Evidence that the axostyle complex generates torque.

A rotary motor in a termite flagellate continually turns the anterior part of the cell (head) in a clockwise direction. Previous descriptive observations implicated the noncontractile axostyle, which runs through the cell like a drive shaft, in the motile mechanism. This study demonstrates directly that the axostyle complex generates torque, and describes serval of its dynamic properties. By laser microbeam irradiation, the axostyle is broken into an anterior segment attached to the cell's head, and a posterior segment which projects caudally as a thin spike, or axostylar projection. Before lasing, both head and axostylar projection rotate at the same speed. After breaking the axostyle, the rotation velocity of the head decreases, depending on the length of the anterior segment. Head speed is not a linear function of axostyle length, however. In contrast, the rotation velocity of the axostylar projection always increases about 1.5 times after lasing, regardless of the length of the posterior segment. Turning the head is thus a load on the axostylar rotary motor, but the speed of the posterior segment represents the free-running motor. A third, middle segment of the axostyle, not connected to the head or axostylar projection, can also rotate independently. No ultrastructural differences were found along the length of the axostyle complex, except at the very anterior end; lenth-velocity data suggest that this region may not be able to generate torque. An electric model of the axostylar rotary motor is presented to help understand the length-velocity data.


Biochemical and immunological analysis of rapidly purified 10-nm filaments from baby hamster kidney (BHK-21) cells.

Juxtanuclear birefringent caps (FC) containing 10-nm filaments form during the early stages of baby hamster kidney (BHK-21) cell spreading. FC are isolated from spreading cells after replating by treatment with 0.6 M KCl, 1% Triton X-100 (Rohm & Haas Co., Philadelphia, Pa.) and DNase I in phosphate-buffered saline. Purified FC are birefringent and retain the pattern of distribution of 10-nm filaments that is seen in situ. Up to 90% of the FC protein is resolved as two polypeptides of approximately 54,000 and 55,000 molecular weight on sodium dodecyl sulfate (SDS) polyacrylamide gels. The protein is immunologically and biochemically distinct from tubulin as determined by indirect immunofluorescence, double immunodiffusion, one-dimensional peptide mapping by limited proteolysis in SDS gels, and amino acid analysis. The BHK-21 FC amino acid composition, however, is very similar to that obtained for 10-nm filament protein derived from other sources including brain and smooth muscle. Partial disassembly of 10-nm filaments has been achieved by treatment of FC with 6 mM sodium-potassium phosphate buffer, pH 7.4. The solubilized components assemble into distinct 10-nm filaments upon the addition of 0.171 M sodium chloride.


Ultrastructural localization of calcium in unfertilized sea-urchin eggs.

The pyroantimonate technique was employed to identify the binding sites for calcium in unfertilized Arbacia punctulata and Strongylocentrotus purpuratus eggs. Since antimony is non-specific and binds with a variety of cations, the indentification of calcium was established by specific chelation with ethyleneglycol tetra-acetic acid (EGTA) and X-ray microprobe analysis. Antimony deposits were observed on the egg's membranes, i.e. plasma, cortical (secretory) granule, pigment granule, smooth-surfaced vesicle, and yolk platelet. Deposits were also observed in the mitochondria, rod-containing vesicles, and the vitelline layer. Two types of yolk platelets were observed: a more numerous electron-opaque platelet which had precipitate along its limiting membrane as well as within the stored-matrix substance, and a less-frequently seen platelet with lower electron opacity which contained precipitate only along its limiting membrane. Deposits were reduced at all sites following exposure of eggs to EGTA either prior to or after osmium-antimonate fixation. Initial fixation in glutaraldehyde followed by postfixation in osmium-antimonate solutions provided better preservation of structure but less precipitation than direct fixation in osmium-antimonate. The organelle sites of calcium binding identified within unfertilized sea-urchin eggs may participate in stimulus-secretion coupling (exocytosis of the cortical granules) and the activation of embryogenesis at fertilization.


Intracellular fibres in cultured cells: analysis by scanning and transmission electron microscopy and by SDS-polyacrylamide gel electrophoresis.

The 3-dimensional structure of the fibrous cytoskeleton of 3T3 cells was examined by scanning electron microscopy of cells extracted with the non-ionic detergent Triton X-100. Detergent-extracted cells consist of the nucleus and an extensive system of fibres, the largest of which correspond to stress fibres visible by phase-contrast microscopy. The system of fibres, which is coterminous with the borders of the native cell, remains firmly adherent to the substratum. The major fibres branch into smaller fibrils which appear to end by ravelling out into fine filaments that constitute a matted network in a plane very close to that of the substratum. In the nuclear region all the major fibres pass over the top of the nucleus, where they may also branch into a system of fine fibrils. Thin-section transmission electron microscopy in conjunction with heavy meromyosin treatment of extracted cells shows the fibres to be composed of native F-actin. Intermediate filaments are also present, and are prominent in the matted network, together with actin filaments. The major proteins of the residue are identified by SDS-polyacrylamide gel electrophoresis as actin, a 56000 Dalton peptide, and histones. Also present are myosin heavy chain, peptides of 225,000 and 250,000, and minor bands at 60,000 and 94,000 Daltons. The non-ionic detergent extracts 70% of the cellular protein, including 50% of the actin and 75% of the myosin. The Triton-insoluble fraction of 3T3 cells appears to constitute, in addition to the nucleus, a stable cytoskeletal system, composed largely of contractile proteins and 10-nm filaments, which functions in maintenance of cell shape, in substratum adhesion, and in positioning the nucleus within the cell.


Direct visualization of the 10-nm (100-A)-filament network in whole and enucleated cultured cells.

Following extraction of actomyosin and tubulin from cultured cells treated with Triton X-100, a cytoskeleton remains which is composed predominantly of the cell nucleus encompassed by a network of 10-nm filaments. After negative staining the dense perinuclear region appears as a densely woven filament net punctuated by patches of high electron density. Enucleation of 3T3 cells with cytochalasin B gives rise to karyoplasts surronunded by 10-nm filaments and cytoplasts in which 10-nm filaments remain situated in the central region of the cytoplasm. While the 10-nm filaments occurred mainly as single filaments in human skin fibroblasts and 3T3 cells, in epithelioid PtK1 and PtK2 cells they were commonly associated in prominent meandering bundles. In addition, in these latter cells after Triton extraction the remaining ribosomes were bound specifically to the 10-nm-filament net. After exposure of 3T3 cells to cytochalasin B the 10-nm filaments formed branches that radiated from the perinuclear region into the immobile cell extensions. Concavalin A had no marked effect on the distribution of the 10-nm-filament net. The results suggest that the 10-nm filaments act primarily as structural elements, serving, in particular, to support and constrain the nucleus in its position in the cell.


Comparative evaluation of a modified zinc sulfate flotation technique.

A modified zinc sulfate flotation technique using Formalinized fecal specimens (F-ZnSO4) was compared to the Formalin-ether (FE) concentration method for the laboratory diagnosis of intestinal parasites. Many laboratories have difficulty storing, using, and disposing of either and need a procedure for concentrating fecal specimens which does not require ether. Comparative data were obtained for the recovery of protozoan cysts and helminth eggs and larvae from feces preserved in Formalin less than and longer than 1 month. Whereas the FE method was found generally to be more efficient, F-ZnSO4 was apparently more effective for the recovery of some species of parasites. F-ZnSO4 was not satisfactory for recovery of schistosome eggs. We conclude that, except for schistosomes, F-ZnSO4 compares favorably to the FE method for detecting infections of clinical significance.


Radioimmunoassay for quantitation of antibodies to alphaviruses with staphylococcal protein A.

A radioimmunoassay (RIA) procedure is described for measuring antibodies to alphaviruses in human and other mammalian sera. The test employed protein Abearing Staphylococcus aureus as a solid-phase immunoadsorbent for (3)H-labeled viruses complexed with immunoglobulin G. Using antibodies produced in humans and guinea pigs, the RIA procedure clearly differentiated among antibodies to Venezuelan, western, and eastern equine encephalomyelitis viruses. Sensitivity of the RIA depended on the concentrations of labeled viruses employed. The dilution of serum that effected binding of 50% of the (3)H-labeled virus (determined by probit analysis) was consistently higher than the neutralizing antibody titer determined by a conventional plaque reduction neutralization test using 80% plaque reduction end points. In addition, sera from 73 individuals were screened for seroconversion following live attenuated Venezuelan equine encephalomyelitis virus vaccine (strain TC-83) inoculation, by RIA using a single serum dilution (1:80); results were identical with seroconversions identified by plaque reduction neutralization test. Hyperimmune Venezuelan equine encephalomyelitis virus sera from a number of mammalian species were successfully titrated by RIA; the species tested were human, guinea pig, white rat, rabbit, burro, dog, monkey, sheep, and cotton rat. The protein A-mediated RIA is a rapid, sensitive, specific, and precise serological tool for measuring antibodies to surface antigens of alphaviruses, and should allow the subsequent development of a competitive binding RIA to measure antigenic potency of inactivated alphavirus vaccines.


Anaerobic and aerobic urethral flora in healthy females.

We characterized the aerobic and anaerobic urethral flora of five healthy females by performing urethral and midstream urine cultures once weekly for 8 weeks. Aerobic cultures were performed monthly for an additional 3 months. Lactobacillus spp. were isolated from 52 of 57 samples, Staphylococcus epidermidis from 42 of 57, Corynebacterium spp. from 26 of 57, and alpha-hemolytic streptococci from 14 of 57. Two subjects had E. coli serogroup O6 and group B streptococci isolated on five occasions, respectively. Anaerobes were isolated from 32 of 35 urethral urines (91%). Bacteroides melaninogenicus accounted for 46% of these isolates. The anaerobic urethral flora varied slightly from week to week, and a similar anaerobic flora was isolated from the introitus, fourchette, and cutaneous perineum. In addition, anaerobes were isolated from 16 of 18 healthy females who had a urethral urine sample cultured once only, and B. melaninogenicus was the most frequent isolate. Of the 21 B. melaninogenicus isolates identified to subspecies, 14 were subsp. intermedius.


Evaluation of prenatal androgen and ovarian secretions on receptivity in female and male rats.

Offspring of rats infected daily from Day 16 through Day 20 of gestation with either 2 mg of testosterone propionate (TP) in .1 ml of sesame oil or oil alone were tested for sexual receptivity following injections of 3.3 microgram of estradiol benzoate (EB) and .5 mg of progesterone (P) beginning at 40, 80, or 120 days of age. At each age, neonatally gonadectomized males and females from TP-injected litters exhibited less receptivity than corresponding oil-injected controls. Prenatally androgenized females were similar to neonatally castrated oil-injected males at all ages. Ovarian implants from birth to 35 days of age significantly increased receptivity in neonatally castrated males and androgenized females. Increasing the age at which testing was initiated systematically reduced receptivity in all groups.


Antibody activity in sera of patients with humidifier disease: studies of the water supply as a source of antigens.

In 5 patients with respiratory symptoms consistent with humidifier or air conditioner disease, a hypersensitivity pneumonitis, no identifiable organism was cultured from the humidifier water (HW). The water source to the humidifier or air conditioner was investigated for possible antigens which might explain the occurrence of the disease. Three subjects who lived in the same city supplied by Lake Michigan had increased IgG antibody activity against the city water supply when compared with another patient from a different city and a control subject. Both of the latter lived in cities with water supplied from Lake Michigan. Two of the 5 patients showed definite inhibition by concentrated tap water (TW) of IgG antibody against HW. The source of antigens in water was not determined but must be considered in cases of this type of hypersensitivity pneumonitis. A sixth case consistent with humidifier disease had Cephalosporium cultured from her HW. On gel diffusion there was a band of identity with her serum against Cephalosporium extract and her own HW, and by radioimmunoassay (RIA) there was over 50% inhibition of her IgG antibody activity against HW by Cephalosporium. No antibody against her city TW was demonstrated.


Nutrient comparison of fresh and field-dried, green-seeded soybeans.

Nutrient composition and biologic utilization of cooked, dried, and ground meals prepared from fresh and field-dried, green-seeded edible soybeans were evaluated. On a dry-weight basis, nutrient content of the fresh and field-dried meals were comparable for protein, fat, calcium, phosphorus, magnesium, copper, and iron; fresh beans tended to have higher zinc content than the field-dried beans. Nutrient values for the green-seeded soybean meals were comparable to published values for full-fat soybean flour. Bioassay results indicated that protein efficiency ratios (PER) for rats fed casein were significantly better than those for the soybean-fed animals. Fresh, green-seeded soybean meal supported significantly better growth than did the field-dried, green-seeded soybean meal. Though significantly lower than that for the reference casein diet, the mean PER for fresh, green-seeded soybean meal was 90 per cent of that obtained with the reference casein. The nutrient analysis and protein bioassay data both indicate that green-seeded soybeans used as a vegetable item in the diet are a potentially significant food source of several important nutrients.


Master standard data quantity food production code. Macro elements for synthesizing production labor time.

Preparation procedures of standardized quantity formulas were analyzed for similarities and differences in production activities, and three entrée classifications were developed, based on these activities. Two formulas from each classification were selected, preparation procedures were divided into elements of production, and the MSD Quantity Food Production Code was applied. Macro elements not included in the existing Code were simulated, coded, assigned associated Time Measurement Units, and added to the MSD Quantity Food Production Code. Repeated occurrence of similar elements within production methods indicated that macro elements could be synthesized for use within one or more entrée classifications. Basic elements were grouped, simulated, and macro elements were derived. Macro elements were applied in the simulated production of 100 portions of each entrée formula. Total production time for each formula and average production time for each entrée classification were calculated. Application of macro elements indicated that this method of predetermining production time was feasible and could be adapted by quantity foodservice managers as a decision technique used to evaluate menu mix, production personnel schedules, and allocation of equipment usage. These macro elements could serve as a basis for further development and refinement of other macro elements which could be applied to a variety of menu item formulas.


Changes of the external and internal pigment pattern upon fertilization in the egg of Xenopus laevis.

External and internal pigment shifts in Xenopus laevis eggs were studied between fertilization and first cleavage. Externally visible, constant features are: (1) the 'activation contraction', a pigment shift towards the animal side taking place between 5 and 15 min post fertilization (p.f.) and (2) the concentration of the pigment around the sperm entrance point leading to the formation of the grey crescent at the opposite side of the egg. Hence, in Xenopus the grey crescent is not formed by rotation of the pigmented cortical layer with respect to the internal egg mass. Histological examination reveals that during the activation contraction the pigment is mainly concentrated in the cortical cytoplasm. Except in the region around the sperm entrance point, from 15 min p.f. onwards, the pigment progressively disperses through the subcortical layer and part of it even moves more deeply into the egg. After fusion of the pronuclei (45--60 min p.f.) the pigment in the subcortical layer forms aggregates. During the pigment shift the yolk-free cytoplasm is displaced dorsally and is ultimately found opposite the sperm entrance point. Thin fibrillar structures in the yolk-free cytoplasm progressively orient themselves parallel to the dorso-ventral plane, and from 40 min p.f. onwards towards the pronuclei. These observations are discussed in connexion with cinematographic observations by Hara, Tydeman & Hengst (1977).