Anterior segment complications of argon laser therapy.

Similar complications have been reported with both xenon arc and argon laser photocoagulation. Emphasis has been placed on complications in the posterior segment. However, with the argon laser, anterior segment complications such as iritis, focal iris atrophy, inequality of pupil size, posterior synechiae, anterior subcapsular and cortical lens opacities and corneal leukomata may occur. The majority of these complications may be prevented by adequate dilatation of the pupil, patient cooperation, and selective use of retrobulbar anesthesia.


The development of corneal astigmatism in contact lens wearers.

In this study of 131 eyes, a comparison of the corneal astigmatism present before wearing hard contact lenses was made with that found after several years of wear (mean length of time: 4.4 years). The eyes were divided into 3 groups according to their ocular rigidity measurements. Statistical analysis of these groups showed that eyes with low ocular rigidity developed a high incidence of with-the-rule astigmatism change and that, furthermore, this change was of significant amount (twice that developed by eyes in the other groups). It was notable that all of the eyes that developed more than 1 1/2 diopters of astigmatism had low ocular rigidity.


Transpupillary photocoagulation of the ciliary processes.

We used transpupillary argon laser to photocoagulate the ciliary processes in 7 eyes of 6 patients in an attempt to reduce intraocular pressure. Only one patient showed a decrease in intraocular pressure to a normal level. This patient was the only one in this series in whom large numbers of ciliary processes could be coagulated. The limiting factor in effective transpupillary argon laser photocoagulation may, therefore, be the total number of ciliary processes visualized and treated.


The surgical correction of refractive errors by keratomileusis and keratophakia.

The methods of performing keratomileusis and keratophakia are described. The principles of the calculations are outlined. Results and complications are enumerated. The indications for the methods are discussed.


Radioimmunoassay of sisomicin.

An antibody induced in rabbits against a gentamicin-bovine serum albumin conjugate cross-reacted with the chemically related aminoglycoside, sisomicin, facilitating the development of a sisomicin radioimmunoassay. Sisomicin was labeled with (125)I using an acylating agent [3-(4-hydroxyphenyl)propionic acid-N-hydroxy-succinimide ester]. The resulting assay produced a linear standard curve on a logit-log plot with a sensitivity of 140 pg. Comparison with a microbiological assay showed no significant difference (P < 0.001) in the measurement of sisomicin by these two methods.


Electrically generated silver ions: quantitative effects on bacterial and mammalian cells.

The inhibitory and bactericidal concentrations of electrically generated silver ions were 10 to 100 times lower than for silver sulfadiazine. Effects on normal mammalian cells were minimal.


Single mitotic center for rodent cochlear duct.

The pattern of terminal mitosis in mouse otocyst observed by Ruben led him to postulate the existence of a growth zone at the junction of saccular and cochlear primordia. With the use of colchicine, an antimitotic drug, a localized zone of mitotic activity has been demonstrated at this predicted site.


Effect of amphotericin B on growth and membrane permeability in Dictyostelium discoideum.

In this study we have determined the effect of the polyene antibiotic amphotericin B on the growth of the eukaryotic microorganism Dictyostelium discoideum. These experiments show that the addition of drug to axenically growing cultures results in an inhibition of growth and cell division. However, with continued incubation, growth is resumed. To determine if the inhibitory effect was due to cell death, the effect of the drug on cell viability was measured. The results showed 10 to 20 times more drug was required to kill cells than to inhibit growth. Since previous studies had indicated that drugs of this type modified cellular permeability, the effect of this drug on osmotic stability of these cells was determined. Results reported in this study show that amphotericin B treatment modifies the cell surface, producing osmotically unstable cells, and that this modification occurs before the onset of cell death and within the same concentration range as used to bring about the inhibition of growth and division. Based on these data it is suggested that the modification in cellular permeability produced by the drug results in the inhibition of growth. This study also reports the results of experiments on the fate of the membrane-damaged cells. These experiments, using radioactive thiourea, showed the restoration of cellular permeability barrier and suggested that the resumption of cell growth occurs after the completion of the repair process.


Ultrastructure of rumen bacterial attachment to forage cell walls.

The degradation of forage cell walls by rumen bacteria was investigated with critical-point drying/scanning electron microscopy and ruthenium red staining/transmission electron microscopy. Differences were observed in the manner of attachment of different morphological types of rumen bacteria to plant cell walls during degradation. Cocci, constituting about 22% of the attached bacteria, appeared to be attached to degraded plant walls via capsule-like substances averaging 58 nm in width (range, 21 to 84 nm). Many bacilli appeared to adhere to forage substrates without distinct capsule-like material, although unattached bacteria with capsules were observed occasionally. Certain bacili appeared to be attached to degraded tissue via small amounts of extracellular material, but others apparently had no extracellular material. Bacilli with a distinct morphology due to an irregularly folded, electron-dense outer layer or layers (about 15 nm thick) and without fibrous extracellular material consituted about 37% of the attached bacteria and were observed to adhere so closely to degraded plant walls that the bacterial shape conformed to the shape of the degraded zone. In the rumen ecosystem, bacteria appeared to adhere to plant substrates during degradation by capsule-like material and by small amounts of extracellular material, as well as by the other means not observable by electron microscopy.


Lipolytic fermentations of stickwater by Geotrichum candidum and Candida lipolytica.

Stickwater, a by-product of the fish meal and oil industry, is an aqueous suspension of fish proteins, lipids, and other materials, and also contains soluble nonprotein nitrogen but not carbohydrate. It is usually partially evaporated by heat to a marketable form called "fish solubles," which is sold with an acid preservative as an animal feed supplement. However, fish solubles are only used to a limited extent in feeds, because the lipids of solubles (averages 11%) are relatively prone to oxidative rancidity development. An investigation was undertaken to digest and/or stabilize lipids in stickwater by lipolytic fermentations and, at the same time, to attempt to increase the protein content as single cell protein. Strains of the yeast Candida lipolytica and the yeast like mold Geotrichum candidum were employed for these investigations. Stickwater fermentations were performed in a laboratory bench top fermentor. Respirometric studies of lipid metabolic activity and microbial observations were periodically performed during these fermentations. Rapid microbial growth and metabolic activity were observed in well aerated cultures. Fermented products were evaluated for chemical composition. Lipid residues were characterized by thin-layer chromatographic procedures. There was evidence of abundant microbial growth, increased lipolytic activity, and decreased lipid content. However, evidence was lacking to show that the protein content of stickwater was actually increased.


Alpha-galactosidase production and use in a hollow-fiber reactor.

Soybean milk serves as a base for a variety of beverages designed for consumption in developing countries. Soybean flour contains raffinose and stachyose considered to be responsible for flatulence often associated with these products (J.J. Rackis, D.H. Honig, D.J. Sessa, and F.R. Steggerda, 1970). alpha-Galactosidase, produced on wheat bran, hydrolyzes the galactooligosaccharides of soybean milk.


Dye reduction method for estimating bacterial counts in ground beef.

A dye reduction method was developed for estimating total aerobic and/or psychrotrophic bacterial counts in ground beef. The method is based on color changes in indicator disks placed on the meat surface.


A mode of selective action of thiadiazolyl urea herbicides.

1,1-Dimethyl-3-(5-tert-butyl-1,3,4-thiadiazol-2-yl)urea(I) was newly synthesized by the authors. (I) was found to have the strongest herbicidal activities among the thiadiazolylurea derivatives and selectivity of (I) was found between barley (tolerant)and wheat (susceptible) plants. Neither absorption by roots nor translocation from roots to shoots of 35S-labeled (I) in barley and wheat correlated with selectively between these species of plants. (I) was metabolized in both species of plants by N-demethylation to 1-methyl-3-(5-tert-butyl-1,3,4-thiadiazol-2-yl)urea(II), and further to non-phytotoxic 3-(5-tert-butyl-1,3,4-thiadiazol-2-yl)urea(III). The rates of the N-demethylating reactions from (I) to (II) and from (II) to (III) were much greater in barley shoots than in wheat shoots, especially in the second demethylation. It was concluded that the selective activity of (I) between barley and wheat plants was mainly due to the difference in rates of N-demethylation of (I). In addition, it was found that cotton plants, well known as tolerant to phenylurea herbicides, were remarkably susceptible to (I). It was indicated that different rates of the N-demethylating reaction between the phenyl and thiadiazolyl herbicides correlated with susceptibility to them.


Radionuclide scanning in the evaluation of Kaposi sarcoma.

A patient with Kaposi sarcoma showed increased uptake of sodium pertechnetate Tc 99m over both clinically evident and occult regions of involvement. The clinically negative but scan-positive regions showed pathologic lesions by biopsy. Gallium citrate Ga 67 and bleomycin sulfate labeled with indium 111 (111In) did not concentrate in the lesions, which suggest that Kaposi sarcoma, despite its resemblance to lymphoma, differs from lymphoma substantially with reference to raionuclide uptake. This report confirms that sodium pertechnetate Tc 99m is the most sensitive agent for diagnostic and therapeutic evaluation of patients with Kaposi sarcoma and suggests taht gallium citrate Ga 67 or bleomycin 111In may be useful in the differentiation of Kaposi sarcoma from lymphoma.


Brain abscess caused by Cladosporium trichoides.

In 17 previously reported cases of cladosporiosis, no reliable therapy was described, and death occurred usually within one year of diagnosis. Pretreatment isolates from our two patients were inhibited by 6.2 mug/ml and 3.1mug/ml of flucytosine, respectively. Although both patients died, postmortem examination results showed that in one patient, the fungus had been eradicated. In the other patient, C trichoides that was isolated at postmortem examination was resistant to flucytosine treatment.


Cystoid maculopathy in diabetics.

Three patients with long-standing diabetes manifested reduced visual acuity and notable bilateral symmetrical cystoid maculopathy, with no other sign of meaningful background diabetic retinopathy, eg, minimal to no microaneurysms, hemorrhages, or exudates. Results of fluorescein angiography showed that the entire retinal capillary bed in the macula was dilated and leaked fluorescein diffusely and profusely, but also showed that there was excellent capillary perfusion. Argonlaser photocoagulation was applied to one eye of each patient in an attempt to resolve the cystoid maculopathy. The laser treatment effectively resolved the edema in the specific areas of treatment, but the foveal and perifoveal areas, which were not treated, showed no areas of resolution of the cystoid edema in two cases and no improvement (and no decrease) in visual acuity in all three cases.


A functional computer recording system for a veterinary diagnostic laboratory.

The establishment of a computer system to process the diagnostic records of the veterinary laboratories of the Queensland Department of Primary Industries is described. A batch system is being used with an optical type input to an International Computers Limited computer. The reports are generated and then reduced photographically to standard size paper. The experience gained and the features and potential of the system are discussed including the particular advantages of the optical input system.


Genetic and environmental components of inconsistency and unrepeatability in twins' responses to a neuroticism questionnaire.

Responses of twins to an 11-item neuroticism scale extracted from questionnaires administered on two widely separated occasions were subjected to a genetic analysis. The results confirm earlier findings concerning the genetic determination of neuroticism and reveal that there is a genetic component in the inconsistency of the test measured by the interaction of subjects and test items. Variation within subjects over the 2-year period between tests was due purely to environmental factors specific to individuals. When a genetic model was fitted to the raw mean squares, there was no evidence that genetic variation was other than additive and no indication of an environmental component common to members of the same family.


Metabolism and structure of the magnesium aspartate-procaine-arrested ischaemic heart of rabbit and man.

The fine structure and the content in energy-rich phosphate compounds, glycogen, and metabolites of the Embden-Meyerhoff-pathway in rabbits hearts or human papillary muscles arrested by magnesium aspartate-procaine are investigated in normothermia and mild or deep hypothermia. In all experimental conditions the break-down of adenine nucleotides and glycogen was distinctly retarded in cardioplegia compared to ischaemic arrest. While e.g. an ATP-content of 3.6 mumole/g wet weight was found after 40 min. at 32 degrees C in the magnesium asparate-procaine arrested heart, it dropped down to 1.3 mumole/g in the ischaemically arrested heart. In cardioplegia after 60 min at 15 degrees C the in vivo contents of ATP and glycogen were determined. The rate in metabolic changes in the magnesium aspartate-procaine arrested human papillary muscle was in the range of that recorded in the arrested rabbit heart. The ultrastructural appearance of the cardioplegically arrested heart did not differ from that of the controls after 20 min at 32 degrees C or 120 min at 15 degrees C. In hearts arrested by cardioplegia 40 min at 32 degrees C first signs of ischaemic lesions e.g. mild swelling of mitochondria and few rarefications in mitochondrial matrix were observed. Because of the significantly improved preservation of the fine structure of the heart and retardation of ischaemically provoked changes in cardiac metabolism, the method of inducing heart arrest by cardioplegia should also clinically be given preference to methods of arresting the heart by ischaemia.


'Isolated' pulmonary valve stenosis as part of more widespread cardiovascular disease.

In 25 patients aged 6 days to 9 years presenting as 'isolated' pulmonary valve stenosis, histology of the myocardium of right and left ventricles, coronary arteries, and ascending aorta has shown abnormality in one or all these areas. Myocardial necrosis, old and recent, unrelated to coronary occlusion was frequent. Myocardial 'dysplasia' involving both ventricles, and resembling hypertrophic cardiomyopathy (HOCM, ASH) was found in 10 and a relation of this to myocardial injury in the fetus is postulated. Varying degrees of coronary occlusion were frequently seen in both right and left coronary arteries. The histology of the ascending aorta was abnormal showing 'higgledy-piggledy' disorder of smooth muscle components in 12 (48%). In a different series of 53 patients who had pulmonary valvotomy for apparent 'isolated' pulmonary valve stenosis there were 14 with clinical evidence of left ventricular abnormality consistent with the pathological changes described, 2 with the same aortic histological changes, and 2 with macroscopical left ventricular hypertrophy. Two of them developed classical hypertrophic cardiomyopathy years later. It is suggested that when pulmonary valve stenosis presents with a thick tricuspid poorly mobile valve, particularly in infants or in patients with evidence of other congenital stigmata, it may be part of a more widespread cardiovascular abnormality. This should be recognized and considered in the evaluation of surgical patients and late survivors who may show unexpected clinical features.


The effect of combined treatment with a platinum complex and ionizing radiation on chinese hamster ovary cells in vitro.

Cis-dichlorobis(cyclopentylamine)platinum II (DBCP) belongs to the group of platinum complexes which have recently been reported to have anti-tumour activity. Its cytotoxic activity in CHO cells is not cell-age-dependent, but enhancement of the effect of ionizing radiation is both dose-and cell cycle phase-dependent. In asynchronous cell populations DBCP acts as a dose-modifying factor for ionizing radiation. Doses of DBCP reducing survival of CHO cells to 26% and 4% applied 1 h before irradiation reduce the Do value of radiation dose-survival curves by factors of 1-34 and 1-59 respectively. In synchronized CHO populations enhancement by DBCP of the effect of radiation is most pronounced in G1 and in late S while it is reduced in mid-S. Possible mechanisms of DBCP-radiation interaction are discussed.


Inhibitory effect of eggs on vitamin B12 absorption: description of a simple ovalbumin 57Co-vitamin B12 absorption test.

Ovalbumin and egg yolks, mixed separately in vitro with radiocyanocobalamin (57 Co-vitamin B12), were served to normal volunteers in a cooked form. Ovalbumin, and to a lesser degree, egg yolks were observed to inhibit vitamin B12 absorption. This observation explains the rather poor assimilation of vitamin B12 from eggs labelled in vivo with 57 Co-vitamin B12.


The determination of volatile fatty acids in the caecum of the conscious rabbit.

1. A method of obtaining dialysed samples from the caecum of the conscious rabbit is described. 2. Values for total volatile fatty acid content and for molar proportions of individual volatile fatty acids in dialysate samples were in good agreement with those obtained from caecal material. 3. The volatile fatty acid level in the caecum throughout the day was determined using two groups of animals, one group fed ad lib. and the other group on a restricted food intake. These results indicated a marked diurnal fluctuation in volatile fatty acid level in the caecum of rabbits fed once/d which was not evident in those fed ad lib.


Characteristics of the association between prothrombin fragment 2 and alpha-thrombin.

The esterolytic activity of bovine alpha-thrombin on the synthetic substrate N-alpha-p-tosyl-L-arninine methyl ester (TosArgOMe) is stimulated when the prothrombin activation fragment, prothrombin fragment 2, is added as previously reported by this laboratory (Heldebrant, C. M., and Mann, K. G. (1973), J. Biol. Chem. 248, 3642). A similar stimulation of beta-thrombin is observed upon addition of prothrombin fragment 2. The binding constant of prothrombin fragment 2 to alpha-thrombin has been determined by the method of Gutfreund ((1972), Enzymes, Physical Principles, Wiley, New York, N.Y., pp 67-71). The dissociation constant is 7.7 X 10(-10)M, and there is one molecule of prothrombin fragment 2 bound per molecule of alpha-thrombin. Prethrombin-2 competes for prothrombin fragment 2, so the enhancement of the esterolytic activity of alpha-thrombin by prothrombin fragment 2 was used as a probe to determine the dissociation constant for the binding of prothrombin fragment 2 to prethrombin 2. The dissociation constant for this association is 1.3 X 10(-10)M. The kinetic parameters for the reaction of alpha-thrombin on TosArgOMe were determined in the absence and presence of prothrombin fragment 2 and are as follows: (a) in the absence of prothrombin fragment 2, Km(app) = 1.92 X 10(-4)M, and k3(app) = 35.8 mol of TosArgOMe/mol of alpha-thrombin s(-1); (b) in the presence of prothrombin fragment 2,Km(app = 1.76 X 10(-4)M, and k3(app) = 60.5 mol of TosArgOMe/mol of alpha-thrombin s(-1). Thus, the stimulatory effect of bovine prothrombin fragment 2 on bovine alpha-thrombin is reflected in k3(app) and not in Km(app). In contrast to the stimulatory effect of prothrombin fragment 2 on the thrombin-catalyzed hydrolysis of TosArgOMe, it inhibits the activity of alpha-thrombin toward N-alpha-benzoyl-L-arginine ethyl ester and N-alpha-benzoyl-L-arginine p-nitroanilide. The inhibition of activity toward these substrates by prothrombin fragment 2 is also reflected in k3(app). Activity toward the nonspecific substrate p-nitrophenyl butyrate was completely inhibited by the addition of prothrombin fragment 2. Prothrombin fragment 2 has no effect on the inhibition of alpha-thrombin activity by the active-site serine inhibitors diisopropyl phosphofluoridate, phenylmethanesulfonyl fluoride, or p-nitrophenyl guanidinobenzoate. Inhibition by the active-site-histidine-modifying inhibitor, N-alpha-p-tosyl-L-arginine chloromethyl ketone, was enhanced by the addition of prothrombin fragment 2. Soybean trypsin inhibitor reduces the stimulation by prothrombin fragment 2, but only at high molar ratios. Prothrombin fragment 2 has no effect on the clotting activity of alpha-thrombin, nor inhibition of this activity by heparin, hirudin, or diisopropyl phosphafluoridate. Bovine prothrombin fragment 2 enhances the esterolytic activity of both human and bovine alpha-thrombin, but human prothrombin fragment 2 does not enhance the esterolytic activity of either human or bovine alpha-thrombin.


Structure and immunogenic behaviour of methylated tobacco mosaic virus.

Tobacco mosaic virus was methylated, using various concentrations of dimethylsulfate. The methylated virus sample with still intact particles was subjected to sequential analysis. The sites and the degree of methylation were determined in the tryptic peptides. Tyrosine 139 and cysteine 27 are more accessible to methylation than tyrosine 72, lysine 68 and tyrosine 2. A limited number of carboxyl groups was also methylated. The ability of methylated and original tobacco mosaic virus to initiate the formation of humoral antibodies and the capacity to induce a delayed-type hypersensitivity reaction were investigated in STU mice. Original tobacco mosaic virus could not induce a delayed-type hypersensitivity reaction but methylated tobacco mosaic virus induced a delayed-type reaction, not depending on whether the virus particles were intact or disintegrated. This phenomenon was strictly linked with the presence of methylester groups.


Microheterogeneity in porcine pancreatic amylase preparations due to disulfide-sulfhydryl exchange.

There are two masked SH groups in pancreatic amylase (EC 3.2.1.1) which become reactive after removal of Ca ions, and then only due to fluctuation of the polypeptide chain. Carboxymethylation of the masked SH groups and isolation of the tryptic CM-peptides have shown that the reacting SH groups in active amylase preparations are not identical, depending on the method of preparation. The microheterogeneity is attributed to an SH-SS exchange reaction taking place during preparation. This exchange results in a mixture of two types of amylase molecules containing different SH groups. The tryptic peptides containing the SH groups have been detected from the radioautogram of the peptide may of the [14C] carboxymethylated protein digest. We identified the SH peptides present in native amylase, and those cysteinyl peptides which form an easily reducible disulfide in the vicinity of these thiol groups, and take part in the intramolecular SH-disulfide exchange.


Age differences in the positional distribution of phosphoglycerides and molecular species of choline phosphoglycerides during development of the chick embryo liver.

The total fatty acid composition and the distribution of acyl groups in the positions 1 and 2 of 1,2-diacyl-sn-glycero-3-phosphocholine and 1,2-diacyl-sn-glycero-3-phosphoethanolamine as well as in the molecular species of 1,2-diacyl-sn-glycero-3-phosphocholine from the chick embryo liver have been investigated at various stages of development. The fatty acid content of the main lipid classes from the yolks was also evaluated. The relative percentages of total saturated fatty acids in 1,2-diacyl-sn-glycero-3-phosphocholine remained constant during liver development; however, the levels of both main saturated acids, palmitic and stearic acids, were reciprocally counterbalanced as the age of the embryo proceeded. On the other hand, arachidonic acid decreased with time whereas linoleic acid and docosahexaenoic acid doubled their proportions during development. The relative amounts of total saturated fatty acids from liver 1,2-diacyl-sn-glycero-3-phosphoethanolamine were also constant during all stages of development; however, palmitic/stearic acid ratio did not change with time in this phosphoglyceride. Fatty acid composition of the yolk phosphoglycerides did not change during embryonic development. It should be noted that marked differences between fatty acid composition of choline and ethanolamine phosphoglycerides from the embryo liver and those of the yolk phosphoglycerides were demonstrated. The general patterns of the positional distribution of the acyl groups in choline phosphoglycerides differed from those of ethanolamine phosphoglycerides. Palmitic and stearic acids were only positioned at position 1 in 1,2-diacyl-sn-glycero-3-phosphoethanolamine whereas both fatty acids did not acylate exclusively the position 1 in 1,2-diacyl-sn-glycero-3-phosphocholine.


Evidence for B-C transition in ultraviolet-irradiated DNA. An infrared linear dichroism study.

Oriented films of calf thymus DNA are studied before and after 254 nm ultraviolet irradiation by infrared linear dichroism. The ultraviolet-irradiated DNA films are changed in their conformation, which is indicated by a replacement of the B-A transition by a B-C transition when the humidity is lowered, from 90% to 75% relative humidity.


Daughter cells as an important factor in determining the physiological state of yeast populations.

Cells of Candida utilis grown in a single-stage chemostat at D = 0.05, 0.1, 0.25, and 0.35 hr-1 were separated into a fraction of scar-bearing mother cells and a fraction of scar-free daughter cells. The scar-free cells were transferred into small batch cultures where the length of the maturation phase, changes in length and width of cells, specific growth rate, and specific rate of RNA and protein synthesis were examined for 5 hr. The daughter cells grown at D = 0.05 hr-1 were very small at the moment of separation from the mother cells (about one-third of the mother cell). Their maturation phase (in a batch culture), at the beginning of which they attain the specific growth rate approaching the mumax of the strain used, lasts for 3 hr. On the other hand, daughter cells grown at D = 0.35 hr-1 are almost the same size as the mother cells at the moment of separation. After transfer to a batch culture they begin to bud almost immediately. Similarly, in their other morphological and physiological parameters they differ strikingly from immature daughter cells which are formed at low specific growth rates. The importance of these differences from the point of view of mathematical modeling of growth processes is discussed.


Innervation of the muscle of the bladder in the rat.

A quantitative ultrastructural analysis of the organisation and terminal density of the nerves associated with the muscle of the body of the bladder was undertaken in the rat. The results obtained were suggestive of the organisation of the intramuscular plexuses into 2 separate and possibly functionally distinct networks. The nerves ramifying within the muscle fascicles mostly contained between 1 and 3 axons and on the basis of counts made in 12 animals, appeared to provide the muscle with only a moderate level of innervation. Although adrenergic terminals were frequently seen in the perivascular plexuses, they were rarely encountered in the nerves within the muscle fascicles. The majority of the terminals in the intrafascicular nerves were similar to those normally classified as cholinergic and, in many, the exposed parts of the axonal membrane were closely related to that of adjacent muscle cells. In addition to clear vesicles and large dense-cored vesicles, many of these terminals contained scattered small dense-cored vesicles. It was suggested that the involvement of the material present in the small dense-cored vesicles with acetylcholine in impulse transmission might account for the atypical features of such transmission in the bladder and in particular its resistance to blockade by atropine. The nerves forming the interfascicular networks contained few axons with cholinergic terminals, and were characterised by much larger numbers of a morphologically distinctive type of terminal. Some of these were indistinguishable from those recently defined as the terminals of purinergic axons but their distribution in the nerves suggests that they represent the terminals of afferent axons and is consistent with the concept that, together with those observed in other tissues, such axons are involved in pain perception.


Human rabies encephalomyelitis.

Seven weeks after he was bitten on the lip by a puppy in the Gambia a patient showed symptons of rabies. Passive and active immunisation was begun three days after the onset of symptons. The evidence indicated that death was a direct consequence of the central nervous system disease rather than any associated complication. Our inability to alter the course of the illness appreciably emphasises the importance of immediate postexposure immunisation in rabies and draws attention to the present lack of effective means of preventing virus replication within the central nervous system.


A case of rabies in man: some problems in diagnosis and management.

A patient who returned from India with a gastrointestinal disturbance subsequently developed the clinical features of rabies. The diagnosis was confirmed by isolation of rabies virus from saliva and the demonstration of rising titres of rabies antibody. He was treated in an intensive care unit, ventilated mechanically, and given rabies antiserum, but cerebral activity ceased and he died after 22 days. His course was marked by numerous bouts of cardiac arrhythmia including complete heart block. This case indicates the need for vaccination against rabies to be carried out immediately after exposure to a possibly rabid dog since encephalitis develops a fatal outcome may not be prevented even with intensive medical treatment.


Change in diurnal temperature rhythm in manic-depressive illness.

The temperatures of six manic-depressive patients were taken every three hours consecutively for many weeks, covering at least one depressive and one manic episode in each patient. While the daily temperature curve was essentially normal in manic phases, with pronounced 24-hour rhythm, during depression the daytime temperatures appeared disorganised, often falling during the morning instead of rising, and with suggestions of a 12-hour rhythm. It may be useful to look on manic-depressive illness as resulting from a desynchronisation of circadian rhythms and to compare the pharmacologies of temperature regulation and mood regulation in psychosis.


Pentobarbital inhibition of progesterone-induced behavioral estrus in ovariectomized guinea pigs.

Administration of pentobarbital inhibits the facilitatory effects of progesterone on the release of gonadotropins. In this experiment facilitatory effects of progesterone on lordosis behavior in guinea pigs were examined with pentobarbital anesthesia. Two major animal groups were subjects: one was short-term ovariectomized (2 weeks) and the other was long-term ovariectomized (several months). All animals received estradiol benzoate (6.6 mug s.c.) followed by progesterone (0.4 mg s.c.) 40 h later. Lordosis behavior was induced by the manual stimulation method of Young et al.29 Sodium pentobarbital (30 mg/kg) was injected 8,4 or 2 h before, simultaneously or 1, 2, 6, or 7 h after progesterone. Animals which received pentobarbital slept for 4.5-5 h with subsequent drowsiness for an additional 0.5-1 h. Pentobarbital injections given 8 h before progesterone had no effect on latency to the first lordosis or on other parameters of estrous behavior. However, pentobarbital delayed the onset of heat in estrogen treated ovariectomized guinea pigs when given 4 h before, 2 h before, or simultaneously with progesterone. The delay was directly related to the length of time the animals remained asleep after the progesterone injection, since estrous behavior was invariably displayed with the latency of controls after the animal awoke. Moreover, in animals which were awake for 1-2 h immediately after the progesterone injection before receiving pentobarbital, the latency of recovery from anesthesia to the first display of lordosis was about 1-1.5 h shorter than in the other pentobarbital groups. In contrast to the latency effects of pentobarbital, the duration of heat was unaffected by the anesthetic for all groups mentioned. In animals which received pentobarbital after they were already in heat, pentobarbital injection terminated heat and abolished it completely, since lordosis behavior was not displayed in the hours after recovery from anesthesia. Gross hypothalamic uptake of progesterone was not influenced by pentobarbital administration. Thus, it is tentatively concluded that an incubation period is necessary for progesterone to mediate the display of estrous behavior in the guinea pig in addition to the time necessary for neural uptake. The way in which pentobarbital interferes with the period of progesterone incubation is not currently known.


Behavioral effects of 5, 7-dihydroxytryptamine lesions of ascending 5-hydroxytryptamine pathways.

In protriptyline (25 mg/kg) pretreated rats stereotactic 5,7-dihydroxytryptamine (5,7-DHT) lesions of the medial plus laternal 5-hydroxytryptamine (5-HE) bundles in the mesencephalon increased the 5-HT fluorescence in these bundles, and reduced the in vitro uptake of [3H] 5-HT in the hypothalamus to 16% of control values after 2 mug 5,7-DHT/4mul and 12% after 4 mug 5,7-DHT/4mul, and in the cortex cerebri to 35 and 34% of control values, respectively. Selective lesion of the medial 5-HT bundle reduced [3H] 5-HT uptake both in hypothalamus and in cortex cerebri to 45-48% of control values, while selective lesion of the lateral 5-HT bundles significantly reduced [3H] 5-HT uptake only in cortex (to 73-75%). No significant change was observed in [3H] noradreanaline uptake after any injection, or in [3H] 5-HT uptake after vehicle injections. Locomotor activity in an open field 3-10 days postoperatively was significantly reduced by lesions of the medial plus lateral 5-HT bundles. 5-Hdroxytryptophan (50 mg/kg) and a peripheral decarboxylase inhibitor (MK 486, 75 mg/kg) 17 days postoperatively induced a pronounced behavioral "5-HT syndrome" in these rats with medial plus lateral lesions but not in controls. Pain sensitivity, as measured by the hot plate test, was not changed by any lesion, even when tryptophan hydroxylase was partly inhibited with alpha-propyldopacetamide (100 mg/kg). Morphine analgesia and acquisition of a one-way avoidance response also were unchanged. Apomorphine (2 mg/kg)-induced locomotor activity and stereotyped behavior, as measured in an Animex activity meter, were not significantly different from control values in the 5,7-DHT groups. It was concluded that the medial 5-JT BUNDLE INNERVATES BOTH THE HYPOTHALAMUS AND THE CORTEX CEREBRI AND THE LATERAL 5-HT bundle mainly the cortex. These ascending 5-HT neurons are involved in maintaining open field ambulation. No wupport was obtained for the view that they are involved in pain mechanisms, in morphine-induced analgesia, in apomorphine-induced motor behavior, or in one-way avoidance learning.


The influence of DL-methionine on the metabolism of S-adenosylethionine in rats chronically treated with DL-ethionine.

The concentration of S-adenosylethionine in the liver of ethionine-fed rats was increased gradually during the process of carcinogenesis. This increase may have been due to the decreased capacity of the treated rats to acetylate ethionine sulfoxide. Ethionine sulfoxide is considered as the main reserve pool of ethionine for the synthesis of S-adenosylethionine. When the ethionine diet was supplemented by DL-methionine (0.3 to 0.9%), the increase in the concentration of S-adenosylethionine during the period of observation (28 to 150 days) was lower and the acetylation of ethionine sulfoxide was significantly higher. The concentration of the total S-adenosyl compounds in the liver of rats on a diet supplemented with DL-methionine was increased over the concentration of S-adenosylethionine in rats fed ethionine alone, and the S-adenosylethionine portion of this fraction was only about 30% lower. The supplementation of the diet with methionine restored the diurnal oscillation of adenosine 5'-triphosphate in the liver, which had been absent in rats ingesting only ethionine.


Microfluoremetric analysis of DNA content changes in murine teratocarcinoma.

The multipotential stem cell of the murine tetratocarcinoma, embryonal carcinoma (EC), is capable of differentiation in vivo and in vitro to nonneoplastic progeny. Undifferentiated EC cells, spontaneously differentiating tetratocarcinoma cells, and differentiated cells derived from EC cells were analyzed for DNA content and chromosome number distributions. Flow microfluorometric and fluorescence cytophotometric analysis of DNA content showed that EC cells had a characteristic diploid (2c) distribution, whereas several differentiated cell lines derived from EC cells had 4c DNA distributions. The tetraploid cell populations studied were capable of cell division but had restricted differentiative potential and were either of low tumorigenicity or non-tumorigenic. In vivo teratocarcinomas, comprised of both EC cells and differentiated cell types, contained diploid and tetraploid populations. Chromosomally, EC cells were neardiploid (39 chromosomes) and differentiated cells were near-tetraploid (62 to 76 chromosomes). The teratocarcinoma provides a model for studying the basic mechanisms that control the growth dynamics of the rapidly and slowly proliferating cell populations present in many tumors.


Kinetic responses of murine sarcoma cells to radiation and hyperthermia in vivo and in vitro.

Cells of a solid mouse mammary sarcoma that can be cultured in vitro and which, upon inoculation, grow in vivo into new tumors, were exposed either in vivo or in vitro to doses of 300 or 600 rads of X-rays and/or to a temperature of 43 degrees for 1 hr. DNA histograms obtained with flow cytofluorometry were sampled at regular time intervals after treatments in order to obtain information on the cells' postexposure kinetics. X-irradiation of exponentially growing cells induced the expected G2 block; heat exposure caused cells to accumulate in S and G2. The sequential treatment (300 rads followed by 1 hr of hyperthermia) resulted in a mitotic delay that was longer than the sum of the delays of the individual treatments. The proliferative behavior of cycling cells in the tumor treated with a dose of X-rays was qualitatively similar to that seen for exponentially growing cells in vitro; however, marked differences were seen after 43 degrees exposure. The heat treatment of tumors in vivo caused a significant decrease in the tumor cell density as compared to the X-ray treatment alone. Sequential X-ray and heat treatment induced a higher fraction of cycling cells than that found in control tumors. However, X-ray or heat treatment alone caused no significant recruitment of resting cells into cycle 1 day after treatment. A model that permits estimation of the fraction of resting cells in a tumor is described.


Modifications and evaluation of double antibody radioimmunoassay of human carcinoembryonic antigen.

Double antibody radioimmunoassay of carcinoembryonic antigen (CEA), a cancer-associated antigen of the human digestive system, was subjected to certain modifications and critically evaluated. Modifications pertained to: (a) the production of a high titer goat anti-CEA antiserum that was rendered highly specific by solid phase immunoabsorption with cyanogen bromide-activated Sepharose conjugates of normal plasma liver, and colon perchloric acid-soluble glycoprotein antigens: (b) the introduction of suitable alterations in the experimental conditions of radioiodination procedure to minimize and to prevent breakdown of the antigen, thus prolonging the storage of the labeled antigen; (c) the extended incubation period of CEA-anti-CEA immune reaction; and (d) the use of sodium acetate buffer, pH 6.1. Furthermore, the use of an automatic pipetting station for accurate and rapid reagent dispensation and statistical analysis of the radioimmunoassay data on a modern computer to ensure strict quality control of the assay provided some definite improvement over the existing assay.


Loss of controlled nuclear division in BHK21 cells passed in vivo.

Low-passage BHK21/C13 cells respond to cytochalasin B (CB) by undergoing limited or controlled nuclear division. These cells respond to CB as normal cells do since nuclear division usually occurs only once. Premature chromosome condensation, a result of mitoses in highly multinucleate cells, occurs in less than 0.5% of the cells. When they are inoculated into weanling hamsters, s. c. tumors appear within 3 to 4 weeks with as few as 10(3) cells. When these cells are returned to cell culture they respond to CB with uncontrolled nuclear division and premature chromosome condensation. All cultured tumors respond in this manner regardless of the number of cells originally inoculated into animals. This suggests at least two possibilities: (a) that loss of controlled nuclear division in BHK cells is closely associated with or required for tumorigenicity, with in vivo passage selecting for a rare tumorigenic variant or (b) that loss of controlled nuclear division is secondary to tumorigenicity and results when cells are passed in vivo, i.e., in vivo passage has the direct effect of causing cells to lose controlled nuclear division. If the first possibility is operating, then it would be expected that a very small fraction of BHK21/C13 cells show uncontrolled nuclear division (approximately 1 of 1000 CB-treated cells). Also, clones of C13 should be nontumorigenic if only 1 of 1000 cells is tumorigenic. Extensive examination of CB-treated C13 cells shows 1 of 1250 cells to be highly multinucleated although not as highly multinucleated as tumor cells. This provides some evidence in support of the first possibility. However, three separate clones of C13 cells were found to be tumorigenic providing evidence supporting the second possibility. BHK21/C13 and various BHK21 tumors all appeared to grow to concentration densities markedly higher than hamster embryo fibroblasts. However, the tumor cells usually grew to the same density as did BHK21/C13 or only slightly higher. This suggests that loss of contact inhibition is not sufficient for loss of controlled nuclear division. It also suggests that the hypothetical 1 of 1000 tumorigenic cells and 1 of 1250 cells with uncontrolled nuclear division do not overgrow the "normal" cells because all cells grow to similar densities. The relationship between the ability to grow in agar and uncontrolled nuclear division was also examined. Approximately 1 of 2500 C13 cells were able to form colonies in agar. All three colonies isolated showed normal control of nuclear division. These results show that the ability to grow in agar may be separate from the expression of uncontrolled nuclear division. They also suggest a fundamental difference between tumor cells and cells that have been grown in agar.


Neuronal and non-neuronal control of the neurosecretory caudo-dorsal cells of the freshwater snail Lymnaea stagnalis (L.).

The cerebral ganglia of the freshwater snail Lymnaea stagnalis contain two clusters of neurosecretory Caudo-Dorsal Cells (CDC). These cells produce a neurohormone which stimulates ovulation. Ganglion transplantation and quantitative electron microscopy show that neuronal isolation of the cerebral ganglia complex (CCC) results in an activation of the CDC. It was, therefore, concluded that the CDC are controlled by an inhibitory neuronal input originating outside the cerebral ganglia. Ultrastructural studies on synaptic degeneration in the CCC suggest that this input reaches the CDC via a special type of synapse-like structure, the type C-SLS. Furthermore, transplantation of CCC into acceptor snails leads to a reduced release and an increased intracellular brekdown of neurohormone in the CDC of the nervous system of the acceptors. It is supposed that these phenomena are caused by the release of an (unknown) factor from the transplanted CCC. Special attention was given to the formation and degradation of a peculiar type of neurohormone granule, the large electron dense granule. The physiological significance of the neuronal and non-neuronal control mechanisms which regulate CDC activity is discussed.


Morphological differences in the interactions between human mononuclear cells and coated or uncoated sheep red blood cells: a freeze-etch study of different types of rosettes.

Enriched preparations of E, EA and EAC rosettes formed by human peripheral blood mononuclear cells were freeze-etched and examined electron-microscopically. In E rosettes only lymphocyte were involved, whereas in EA and EAC rosettes lymphocytes and mononuclear phagocytes participated as rosette-forming cell. In EA and EAC rosettes, cytoplasmic extensions of the rosette forming cell were seen to penetrate the sheep red blood cell, whereas E rosettes showed a broad zone of adherence without penetration. None of the three types of rosettes showed an interspace between the membranes. Unlike E rosettes, EA and EAC rosettes showed polarity in the adherence of sheep red blood cells. These observations made by freeze-etch electron microscopy indicate distinct morphological differences between rosettes formed with coated or uncoated erythrocytes.


Effects of acetylsalicylic acid on the ductus arteriosus and circulation in fetal lambs in utero.

Intra-arterial and intravenous catheters were inserted in six fetal lambs at 125-130 days of gestation. On the following day, fetal arterial pressures and blood gases were monitored and fetal cardiac output and its distribution were measured by injection of radionuclide-labeled microspheres 15 mum in diameter. Acetylsalicylic acid, 55-90 mg/kg of estimated fetal weight, then was administered into the fetal stomach. Fetal pulmonary arterial pressure rose significantly after an average of 58 minutes, increasing the pressure difference between the pulmonary artery and the aorta from 2 +/- 0.3 (SEM) mm Hg during control to 11.2 +/- 1.6 mm Hg. Resistance across the ductus arteriosus rose from 4.2 +/- 0.5 (SEM) to 27.4 +/- 4.01 units, and flow fell from 495 +/- 44 (SEM) to 409 +/- 20 ml/minute. The proportion of combined ventricular output distributed to the placenta, adrenals, heart, and lungs increased, whereas the proportion of combined ventricular output distributed to the brain, liver, intestine, kidneys, and upper and lower body fell. In two fetuses infusion of prostaglandin E1 reversed the pulmonary hypertension. Inhibition of prostaglandin synthesis in fetal lambs produced constriction of the ductus arteriosus and redistribution of cardiac output. It is probable that prostaglandins, particularly E1, are involved in regulation of blood flow through the ductus arteriosus and various vascular beds in the normal resting fetus.


Simultaneous determination of LH activity in blood serum of sheep during three synchronized oestrous cycles by Bio and Ria method.

LH activity has been in 6 Merino sheep by means of the biological (Bio) and radioimmunological (RIA) test during three subsequent synchronized oestrous cycles. The vaginal sponges were impregnated with 30 mg chlormadinone acetate. The results obtained by the two afore mentioned methods are similar. Oestrous appeared in the average within 96 hours after removal of the sponges. The first increase of LH activity (Bio) was found to occur within 48--72 hours after removal of the sponges (270--400 IU/100 ml serum), the second increase within 15--16 days (450-570 IU/100 ml) and the third within further 16 days (390--590 IU/100 ml serum). The values obtained in the first oestrous cycle by RIA showed an LH peak within 62--110 hours after removal of the sponges (75--90 ng/ml in the second oestrous cycle 86--135 ng/ml and in the third one 58--60 ng/ml serum. In three animals no LH peak could be observed during the first, second or third oestrous cycle. The failure to prove LH peaks in three sheep during different phases of the oestrous cycles is explained by the fact that the synchronization led to heat which, however, did not result in fertile ovulation.


Developmental toxicity in the rat after ingestion or gavage of organophosphate pesticides (Dipterex, Imidan) during pregnancy.

The structural development of fetuses was altered when Dipterex was administered by diet to pregnant rats from days 6 through 15 of gestation. Major external and skeletal alterations occurred after consumption of 432 or 519 mg/kg body weight per day, only minor skeletal changes occurred in the 375 mg/kg dose group and the incidence of alterations in the 145 mg/kg dose group was not significantly different from that in the pair-fed controls. The malformations seen at the two highest doses did not result directly from the associated decrease in food consumed. Dipterex was not shown to have teratogenic potential when given for the same time span, once daily by gavage, even at levels that produced maternal lethality. Imidan was not teratogenic when similarly given, either by diet at concentrations that resulted in a 45% reduction in food consumption, or by gavage at dose levels that resulted in some maternal lethality. Data collected from pair-fed control females revealed that limitation of food consumption to 13--15 g/rat per day from days 6 through 15 of gestation did not result in increased fetal mortality or stunting. However, fetal weight was reduced slightly, and the incidence of minor skeletal changes was approximately three to four times that among fetuses of control dams that were not pair-fed.


Status of marine biomedical research.

A meeting on Marine Biomedical Research, sponsored by the National Institute of Environmental Health Sciences (NIEHS), National Institutes of Health and the Smithsonian Institution Museum of Natural History, was attended by approximately 125 scientists, directors and representatives from many of the country's marine biological laboratories, and government agencies whose interests and responsibilites are in the marine biology and health areas. The purpose of the meeting was to explore the undeveloped research opportunities in the area of marine biology for the advancement of our understanding of human health problems and to provide information on the current status of marine biology laboratories. The meeting was devoted to presentations and discussions in four general areas: (1)Marine Species as Models for Human Disease; (2)Environmental Carcinogenesis and Mutagenesis; (3)Human Health and the Marine Environment--infectious agents and naturally occurring and foreign toxins; and (4)Drugs from the seas. Representatives from twelve of the country's approximatley 40 marine laboratories discussed their organization, developmental history, scientific programs, facilities, and present status of their support. The presentations served as a background and stimulated very lively analytical and constructive discussions of the undeveloped research and education potential residing in the marine environment and biological laboratories for a better understanding of many human health problems; some scientific areas that should be developed to realize this potential; and the needs and problems of marine laboratories that require attention and support if they are to survive and realize their possibilities.


[Herpes simplex encephalitis in newborns. Treatment with 5-iodo-2'-deoxyuridin? (author's transl)].

Premature twins both affected with generalized herpes simplex infection including CNS involvement were treated with 5-Iodo-2'-deoxyuridin (IDU). Therapy was started 6 days and 5 days, respectively, after appearance of the first symptoms. Nevertheless both babies died, twin A at the age of 24 and twin B at the age of 28 days. Herpes virus was cultured from all organs in twin A. The literature shows a decrease in mortality in children treated with IDU for herpes simplex encephalitis. The percentage of survivors with ultimate severe CNS damage, however, is very high and remains in treated or untreated patients the same. The institution of IDU therapy in neonates with manifest HSV encephalitis is therefore not unequivocally justified.


Heart norepinephrine concentration after chronic alcohol ingestion in the rat.

The effect of long-term alcohol ingestion on the norepinephrine concentration of the heart was investigated in rats. The alcoholic animals showed a highly significant increase in cardiac norepinephrine concentration as compared with the corresponding controls. It is further suggested that continued exposure to high levels of norepinephrine may play a role in the development of cardiomyopathy in chronic alcoholism.


In vitro fertilization of mouse and hamster eggs after freezing and thawing.

Mouse eggs (12-14%) and hamster eggs (87-94%) appear normal upon thawing after having been stored for 30 min to 25 h at -70 degrees C and -196 degrees C. While 89-98% of the normal appearing hamster eggs are capable of fertilization in vitro, only 4-13% of the mouse eggs can be fertilized. The ability of fertilized eggs to develop in culture and their viability upon transfer to pseudopregnant recipients is under investigation.


Increased plasma creatine kinase activity in rabbits: effect of systematically repeated blood sampling.

Various physical, chemical and biological factors are involved in an increased plasma creatine kinase activity. Repeated blood sampling induced in all rabbits studied a reaction of similar pattern but of different intensity, expressed by a maximum of plasma CPK activity. The physiological origins of this variation of CPK activity seems to be, as seen in control animals, the consequence of emotional stress due to handling and possibly an additional stress due to the venepunctures.


Effect of 6-hydroxydopamine pretreatment on spontaneous convulsions induced by barbital withdrawal.

Following withdrawal from chronic barbital administration, 6-hydroxydopamine pretreated rats show a greater number and an earlier onset of spontaneous convulsive seizures than do rats pretreated with the saline-ascorbic acid vehicle.


Suppression of ethanol withdrawal by dopamine.

An ethanol-inhalation technique was used to determine a potential relationship between dopamine and central nervous effects produced by alcohol. Both L-DOPA and intracranially injected dopamine resulted in attenuation of ethanol-induced withdrawal convulsion scores, whereas, haloperidol, a known dopaminergic blocker was found to significantly increase convulsion scores.


Results of the first 1000 radioreceptorassays for the determination of human chorionic gonadotropin: a new, rapid, reliable, and sensitive pregnancy test.

One thousand radioreceptorassays for human chorionic gonadotropin (hCG) for the diagnosis of pregnancy were analyzed under eight categories. The high sensitivity (50 pg/ml) and specificity, coupled with performance time of the assay (1 hour), provided a new modality for the early diagnosis and management of normal and abnormal pregnancy in the first trimester. The radioreceptorassay becomes positive 1 week after conception. In the normal pregnancy this assay has been 100% correct at the time of the first missed menses. All ectopic pregnancies demonstrated positive tests, and no false negatives occurred. On the other hand, a negative assay excluded ectopic pregnancy.


In vitro biosynthesis of rabbit blastokinin.

The authors have incubated, in vitro, estrous rabbit endometrium with 3H-leucine and pseudopregnant rabbit endometrium with 14C-leucine. By the double-labeling technique and the use of the 14C:3H ratio, it was shown that blastokinin (BKN) is neosynthesized in vitro and that this neosynthesis probably also affects other protein components. By ion exchange chromatography of the supernatant of the pseudopregnant endometrium and previously purified BKN, it was possible to confirm the in vitro biosynthesis and to exclude interfering phenomena. Time dynamics showed that biosynthesis was more marked between the 18th and 24th hours of incubation.


Cell proliferation kinetics in normal human gastric mucosa. Studies on diurnal fluctuations and effect of food ingestion.

Cell proliferation of human gastric mucosa was studied in 3 healthy subjects. Biopsies were obtained from the antral and fundic part of the stomach through a fiber gastroscope. After in vitro incubation with tritiated thymidine, autoradiographs were prepared by the dipping technique. Labeling and mitotic indices were estimated. The number of DNA-synthesizing cells and the mitotic frequencies were higher in antral mucosa compared to mucosa in the fundic part of the stomach. Biopsies were taken after 12 hr of fasting and at various intervals during the following day. It was not possible to demonstrate any diurnal variations in the labeling and mitotic indices. Furthermore, biopsies were taken at various intervals after ingestion of a protein-rich meal, preceded by 12 hr of fasting. Significant changes in cell proliferation were not found either in antral or in fundic mucosa, indicating that a meal does not affect the rate of cell renewal in human gastric mucosa.


[Can cardiotocography provide information regarding fetal death in utero? Casuistic (author's transl)].

A case of fetal death in utero is discussed in which fetal heart tones were picked up with the cardiotocograph via direct leads from the scalp electrode during labor induction, i.e., during the course of labor. After delivery, however, it was apparent that the fetus had been dead for some time (third grade maceration). Technical errors originating in the instrument itself could be excluded since the instrument was used successfully for other deliveries before and after this particular birth. A defect caused by the scalp electrode was also not possible since the electrode was changed three times.


Genetic and morphological study of aggregation in the cellular slime mold Polysphondylium violaceum.

A system for genetic analysis in the cellular slime mold P. violaceum has been developed. Two growth-temperature-sensitive mutants were isolated in a haploid strain and used to select rare diploid heterozygotes arising by spontaneous fusion of the haploid cells. A recessive mutation to cycloheximide resistance in one stain enables selection of segregants, which often appear to be aneuploid.-Aggregation-defective (ag-) mutants having a wide range of phenotypes were isolated in both temperature-sensitive strains after nitrosoguanidine treatment, and complementation tests were performed between pairs of these mutants. Of 380 diploids isolated, 32 showed defective aggregation and were considered to contain 2 noncomplenting ag- mutations. Among noncomplementing mutants interallelic complentation is common. Noncomplementing mutants fall into 4 complementation groups, and those within each complementation group are phenotypically similar. Statistical analysis of the results suggests that the number of complementation units involved in aggregation is about 50.


[Jactstio capitis--motor stereotypy in rodents].

It is found that jactatio capitis, a stereotyped hyperkinesis in the form of pendulum-like motions of head and body, reported earlier by other authors as provoked by different neurotropic drugs, can appear in rats and mice spontaneously under emotional excitement. This hyperkinesis has different frequency in different strains, and the frequency and the degree of expression increase along with the increase of the degree of inbreeding. Hybridological analysis has shown that the jactatio capitis is connected with the suppression of eumelanin synthesis, and is a manifestation of pleiotropic effect of genotypes c or pp for C and Ploci of coat colour. It occurs in albinos and in animals of yellow colour, and is never found in those with black, brown or grey colour, or in black-, grey- or brown-spotted animals. The jactatio capitis shows a negative phenotypical correlation with emotionality as determined by the number of defecations, and has a positive connection with the predisposition to audiogenic seizures. The jactatio capitis seems not to be serotonin-ergic symptom. As the stereotypic movements are combined with a kind of "freezing", the authors consider the jactatio capitis to be a kind of catatonic syndrome.


[Isolation and characteristics of temperature sensitive mutants in a culture of Chinese hamster cells].

The treatment of a temperature-resistant glutamine-independent clone of Chinese hamster cells (237 glu+ tr) with 5-bromodeoxyuridine (BUdR) for 48 or 72 hours at 40 degrees C and subsequent illumination with visible light was used for selection of spontaneous and N-methyl-N'-nitro-N-nitrosoguanidein-induced ts mutants. 30 clones were isolated. Their temperature-sensitivity was studied at two cell densities: 500 and 10000 (or 15 000) cells per dish. 21 clones proved to be temperature-sensitive. The expression of temperature-sensitivity of 11 clones was independent of cell densities used. The remaining 10 clones could be divided in two groups: those expressing temperature-sensitivity only at low plating inoculum (6 clones) and those temperature-sensitive at 10 000 (15 000) plated cells per dish (4 clones). Possible mechanisms of this phenomenon are discussed. Most isolated clones behaved as leaky mutants. Isolation of ts clones without special mutagen treatment from a tr cell population after its permanent incubation at 40 degrees C, restrictive for ts cells, is regarded as strong evidence in favour of the mutagenic effect of BUdR and visible light.


The reaction of the Dolichos biflorus lectin with non-human bloods.

The reaction of the Dolichos biflorus lectin with a series of animal bloods is reported. The red cells of 4 species were found to agglutinate from among the 37 different animal species tested.


Trisomy 9p resulting from maternal 9/21 translocation.

The clinical picture found in a child with trisomy 9p confirmed that this chromosomal syndrome is a entity, which arises from maternal translocation t(9;21).


Correlations between immunoglobulin- and antibody-synthesizing cells during primary and secondary immune responses of rats immunized with peroxidase.

The development of cells synthesizing immunoglobulins without detectable antibody activity and of antibody-synthesizing cells was studied during primary and secondary immune responses of rats immunized with horseradish peroxidase. After primary immunization with peroxidase emulsified in Freund's complete or incomplete adjuvant, the first antibody-producing cells appeared 4 days after injection. They were preceded by cells synthesizing IgG and IgM without antibody function, appearing 3 days after giving antigen. The ratio between the latter and the former population of cells regularly decreased during the primary response. Seventy to 100 per cent of cells synthesizing immunoglobulins without antibody activity were induced by the antigen, the remainder being induced by the adjuvant. In both populations, the positive cells were always immature or mature plasmocytes. At various times after primary injection, animals received a booster inoculation of soluble peroxidase or of peroxidase emulsified in Freund's adjuvant. Antibody-producing cells, in early stages of differentiation, appeared between 2 and 3 days after challenge and were not preceded by cells synthesizing immunoglobulins without antibody function. These latter cells were reduced or absent after secondary challenge. Increasing the sensitivity of detection of active sites of antibodies, by using direct methods of staining with fixed or unfixed cells gave no increase of antibody-producing cells.


Stress and the immune response in rats.

The in vitro response of sensitized splenic lymphocytes to antigen (thyroglobulin) was increased by crowding and decreased by isolation in female rats. Both isolated and crowded male rats responded by a decrease in the in vitro reactivity of lymphocytes to antigen. The response of the lymphocytes to PHA was not altered in any consistent manner. Similar animals, both control and those immunized with thyroglobulin, were tested for an effect of in vivo injections of epinephrine on the in vitro reactivity of lymphocytes; epinephrine was given intraperitoneally 30 min before the rats were killed for removal of spleens. Incorporation of 3H-thymidine by lymphocytes was greater in control cultures (neither PHA nor antigen present) but there was a decreased response to either PHA or antigen when epinephrine had been injected.


Aflatoxin occurrence on raw and cooked york soybeans inoculated with three aspergillus isolates.

Raw and cooked soybean media were inoculated in separate experiments with Aspergillus flavus NRRL 3251, A. flavus ATCC 15548, and A. parasiticus NRRL 2999 isolates. The toatl quantity of aflatoxins produced and the percentage distribution of aflatoxins B1, B2, G1, and G2 varied with the state of the medium (raw or cooked) and with the fungal isolate used. Cooked soybean medium supported higher aflatoxin productions by A. flavus NRRL 3251 and A. parasiticus NRRL 2999 than did raw medium. Larger quantities of aflatoxins B1, B2, G1, and G2 were produced by A. flavus ATCC 15548 on raw soybean medium than were produced by any of the isolates on cooked medium. Application of these data is discussed briefly in relation to possible use of soybean media as a source of aflatoxin production.


Repolymerizability of native thin filaments of rabbit skeletal muscle.

The repolymerizability of native thin filaments of rabbit skeletal muscle was examined under various conditions. The native thin filaments usually did not repolymerize after depolymerization, but it was found that the process was only inhibited at the nucleation step of the G-F transformation of actin. The inhibited state was released by the addition of sonicated F-actin fragments as seeds or by direct sonication. An attempt to isolate the actin moiety from native thin filaments showed that the isolated actin was very similar to Straub-type G-actin2 in its polymerizability. However, a protein factor which inhibits the polymerization of G-actin was obtained by ammonium sulfate fractionation of the depolymerized native thin filaments. Thus, the apparent lack of repolymerizability of native thin filaments was found to be due to the action of this factor on the polymerization of G-actin.


Intracellular retinol-binding proteins from bovine pigment epithelial and photoreceptor cell fractions. Purification of high molecular weight lipoglycoproteins.

Cytosol retinol (vitamin A)-binding lipoglycoproteins were isolated from preparations of bovine retinal pigment epithelial cells and from rod photoreceptor outer segment fractions. The lipoglycoproteins had molecular weights of at least 1.5 X 10(6) as determined by gel filtration chromatography. The purified lipoglycoprotein which was obtained from pigment epithelial cells contains at least two kinds of polypeptide subunits (molecular weight about 35,000 and 150,000), while the rod outer segment cytosol lipoglycoprotein contains at least four kinds of polypeptide subunits (molecular weight about 50,000, 75,000, 120,000, and, 200,000). The pigment epithelial cytosol retinol-binding material contains about 30% (by weight) of lipid, while the material obtained from rod photoreceptor outer segment fractions contains about 65% (by weight) of lipid. Both lipoproteins contain predominantly cholesterol and phosphatidylcholine, while cholesterol esters, triglycerides, phosphatidylethanolamine, and phosphatidylserine are absent. The amino acid compositions of the delipidated proteins from pigment epithelial cells and rod outer segment fractions are similar. The neutral and amino sugars are co-valently linked to the polypeptide moiety of the molecule. The absorption and circular dichroism spectra of the lipoproteins obtained from pigment epithelial cells and rod outer segment fractions are different.


Regulation of malic enzyme synthesis by insulin triiodothyronine, and glucagon in liver cells in culture.

Cells isolated from the livers of 17- to 19-day-old chick embryos were maintained in a chemically defined culture medium. During 3 days in culture the activity of malic enzyme, a measure of its concentration, was stimulated 2-, 23-, or 77-fold by insulin, triiodothyronine, or insulin plus triiodothyronine, respectively. Glucagon blocked the stimulation caused by insulin plus triiodothyronine. Changes in the relative synthesis of immunologically isolated malic enzyme were similar in magnitude and direction to the changes in enzyme activity. Degradation of malic enzyme was unaffected by the three hormones. Both soluble protein and malic enzyme were degraded with a t1/2 of about 30 hours. In cells preincubated for 2 days with insulin, synthesis of malic enzyme was stimulated 4.5-fold within 3 hours after adding triiodothyronine and reached an apparent new steady state after 24 to 30 hours. If the rate of enzyme synthesis was dependent on the concentration of cytoplasmic malic enzyme messenger RNA, then this messenger RNA appeared to have a t1/2 of about 18 hours. Glucagon rapidly and specifically inhibited synthesis of malic enzyme in preinduced cells, suggesting an action at the level of translation or cytoplasmic messenger RNA processing.


Synthesis and degradation of folate reductase in sensitive and methotrexate-resistant lines of S-180 cells.

The methotrexate-resistant AT-3000 line of S-180 cells has at least 150-fold more immunologically cross-reactive folate reductase than sensitive cells. Highly specific immunologic and protein purification procedures were used to show that the increased enzyme levels in this line are due to a corresponding increase in the rate of folate reductase synthesis. This observation indicates that the relative turnover of the enzyme is not significantly different in the two lines. Folate reductase was purified to homogeneity from both the sensitive and the methotrexate-resistant cells. Comparison of various physical, kinetic, and immunochemical properties of the enzymes revealed no differences. These observations suggest that the AT-3000 line contains one or more regulatory variations leading to the over-production of folate reductase protein that is similar, if not identical, to that produced by sensitive cells. In resistant cells, specific immunoprecipitation experiments demonstrated that folate reductase comprises as much as 7 to 8% of the continuously labeled soluble protein and 6 to 7% of the soluble protein synthesis. Growth of these lines in the absence of methotrexate resulted in a slow decrease in the level of folate reductase to less than 1%. This decrease corresponded to a similar decrease in the relative rate of enzyme synthesis. Variations in the level of folate reductase with cell growth are also due to changes in the relative rate of enzyme synthesis. In the AT-3000 line, pulse decay experiments showed that the half-life of folate reductase was long (50 hours) relative to cell doubling time (24 hours), and also that methotrexate had little or no effect on the turnover of the enzyme. Comparison of the incorporation of radioactive leucine into folate reductase in continuous and pulse labeling experiments gave independent confirmation of these results. Therefore, the relative rate of folate reductase synthesis was the major parameter determining the amount of folate reductase under all examined conditions that resulted in altered levels of the enzyme in resistant cells.


Altered regulation of the rate of synthesis of dihydrofolate reductase in methotrexate-resistant hamster cells.

Dihydrofolate reductase was isolated from hamster cells sensitive to methotrexate and from a methotrexate-resistant subline which has an elevated enzyme level. The enzymes were compared by peptide map analysis, and no differences in primary structure could be detected. The rates of enzyme degradation and synthesis were determined in both cell lines by a novel approach based on the enzyme specific radioactivity (called radioaffinity labeling). Degradation of reductase was minimal in both cell lines, whereas the rates of synthesis were directly proportional to the steady state concentrations of enzyme. Thus the resistant cells synthesized reductase at a rate which was 140 times faster than that in sensitive cells. Therefore the high concentration of dihydrofolate reductase in the methotrexate-resistant cells is probably the result of an alteration of a cellular component which control the synthesis of the enzyme.


Sperm aster in rabbit zygotes: its structure and function.

Microscope observations of rabbit zygotes demonstrate that a sperm aster forms in association with the male pronucleus approximately 1 h postinsemination and consists of two regions. One, the centrosphere, contains a dense aggregation of cisternae of smooth endoplasmic reticulum and microtubules. The second consists of fascicles of microtubules which emanate from the centrosphere. Fertilized rabbit eggs were cultured in medium containing colcemid in order to determine its effects on various events of fertilization, such as movements of the male and female pronuclei and DNA synthesis. No evidence was obtained to indicate that a sperm aster is formed in colcemid-treated zygotes. In addition, migration and close apposition of the pronuclei do not take place. Breakdown of the pronuclear envelopes and condensation of the maternally and paternally derived chromosomes occur even though the pronuclei fail to migrate centrad. Autoradiographic analysis of the synthesis of DNA by both pronuclei demonstrates that their migration into close apposition to one another is not required for the incorporation of tritiated thymidine.


Properties of tubulin in unfertilized sea urchin eggs. Quantitation and characterization by the colchicine-binding reaction.

The colchicine-binding assay was used to quantitate the tubulin concentration in unfertilized Strongylocentrotus purpuratus eggs and to characterize pharmacological properties of this tubulin. Specificity of colchicine binding to tubulin was demonstrated by apparent first-order decay colchicine-binding activity with stabilization by vinblastine sulfate, time and temperature dependence of the reaction, competitive inhibition by podophyllotoxin, and lack of effect of lumicolchicine. The results demonstrate that the minimum tubulin concentration in the unfertilized egg is 2.71 mg per milliliter or 5.0% of the total soluble cell protein. Binding constants and decay rates were determined at six different temperatures between 8 degrees C and 37 degrees C, and the thermodynamic parameters of the reaction were calculated. delta H0=6.6 kcal/mol, delta S0=46.5 eu, and, at 13 degrees C, delta G=-6.7 kcal/mol. The association constants obtained were similar to those of isolated sea urchin egg vinblastine paracrystals (Bryan, J. 1972. Biochemistry. 11:2611-2616) but approximately 10 times lower than that obtained for purified chick embryo brain tubulin at 37 degrees C (Wilson, L.J.R. Bamburg, S.B. Mizel, L. Grisham, and K. Creswell. 1974. Fed Proc. 33:158-166). Therefore, the lower binding constants for colchicine in tubulin-vinblastine paracrystals are not due to the paracrystalline organization of the tubulin, but are properties of the sea urchin egg tubulin itself.


The germinal vesicle nucleus of Xenopus laevis oocytes as a selective storage receptacle for proteins.

The amorphous nucleoplasm of the germinal vesicle nucleus of Xenopus laevis oocytes has been selectively extracted under conditions which leave the nuclear formed elements morphologically intact. The nucleoplasm contains about 97% of the total nuclear proteins and on SDS-polyacrylamide gels some 68 polypeptides can be distinguished. On the basis of solubility differences, the nucleoplasmic proteins can be classified into two categories. The first consists of soluble or easily solubilized proteins which comprise about 34 polypeptides making up 87% of the nucleoplasm. A few of these proteins show electrophoretic mobilities similar to those of soluble proteins of the cytoplasm, but most are unique to the nucleus. The residual 13% of the nucleoplasmic proteins are tightly bound to a nucleoplasmic gel and can be extracted only by solubilizing the gel. The solubility characteristics of the proteinaceous gel suggest a complex held together by salt, nonpolar, hydrogen, and possibly disulfide bonding. Some 34 polypeptides can be distinguished in this gel fraction, including prominent and highly enriched polypeptides of about 115,000 and 46,000 daltons. The relatively soluble fraction of the nucleoplasm does not contain informofers and contains little or no nucleic acid. Evidence is presented that if histones are present in the germinal vesicle, they can comprise no more than about 8% of the total protein. The possibility is discussed that the unique polypeptides of the nucleoplasm may be sequestered there by selective adsorption to or in the nuclear gel.


Steroids and hematopoiesis. II. The effect of steroids on in vitro erythroid colony growth: evidence for different target cells for different classes of steroids.

Androgenic steroids and their non-androgenic 5beta-H metabolites enhance the number of colonies of hemoglobin synthesizing cells grown from rat bone marrow in response to a standard (0.25 unit/ml) concentration of erythropoietin. The target cells for two steroids were found to be different. Cells influenced by the androgen, fluoxymesterone (fluoxy), resembled cells responding to erythropoietin in their cycle characteristics, as measured by tritiated thymidine suicide, and in their physical characteristics, as determined by velocity sedimentation gradient separation. Cells responding to etiocholanolone (etio) had a much lower tritiated thymidine suicide rate and different sedimentation velocities. Preincubation of marrow cells with etio for two hours was sufficient to enhance erythroid colony growth by 84%, whereas a similar incubation with fluoxy produced no increment. These studies demonstrate that different classes of steroids may influence in vitro erythropoiesis by acting on distinct populations of marrow cells. Fluoxymesterone appears to act through cells already committed to respond to erythropoietin, while etiocholanolone appears to act on a separate, perhaps more primitive population of marrow cells.


Studies on the intranuclear distribution of human and mouse genomes and formation of human-mouse hybrid cells.

Autoradiographic examination of early hybrid cells formed by the fusion of 3H-thymidine labeled D98/AH2 cells and 3T3-4E cells revealed that human and mouse chromosomes are often separated within metaphase and interphase nuclei. Although the marked separation of human and mouse chromosomes progressively disappeared with succeeding cell division, the occurrence of sectored nuclei in 16-cell hybrid colonies and the labeling pattern of human chromosomes within separated metaphases indicate that separation of human and mouse chromosomes may persist through several mitoses. Fusion of 3H-thymidine labeled D98/AH2 cells and 3T3-4E cells was coupled with aminopterin selection to study other aspects of hybrid cell formation. Hybrid cells arose from heterokaryons by cell division. Nuclear fusion may have occurred in two out of several thousand heterokaryons examined. However, these could reflect the close apposition of adjacent nuclei. A large fraction (greater than 0.5) of D98/3T3 heterokaryons underwent at least one cell division. However, the number of hybrid colonies containing more than eight cells at ten days following fusion was about 0.03 of the total number of heterokaryons. Many hybrid colonies arrested growth before the 8-cell stage, and the cells in such colonies exhibited nuclear abnormalities.


Human epidermal growth factor and the proliferation of human fibroblasts.

The effect of human epidermal growth factor (hEGF), a 5,400 molecular weight polypeptide isolated from human urine, on the growth of human foreskin fibroblasts (HF cells) was studied by measuring cell numbers and the incorporation of labeled thymidine. The addition of hEGF to HF cells growing in a medium containing 10% calf serum resulted in a 4-fold increase in the final density. The presence of hEGF also promoted the growth of HF cells in media containing either 1% calf serum or 10% gamma globulin-free serum. The addition of hEGF to quiescent confluent monolayers of HF cells, maintained in a medium with 1% calf serum for 48 hours, resulted in a 10- to 20-fold increase in the amount of 3H-thymidine incorporation after 20-24 hours. The stimulation of thymidine incorporation was maximal at an hEGF concentration of 2 ng/ml, was dependent on the presence of serum, and was enhanced by the addition of ascorbic acid. In confluent cultures of HF cells, subject to density dependent inhibition of growth, hEGF was able to stimulate DNA synthesis more effectively than fresh calf serum. Human EGF stimulated DNA synthesis in quiescent cultures, however, regardless of cell density. The addition of rabbit anti-hEGF inhibited all effects of this growth factor on HF cells.


Separation of megakaryocytes from rat bone marrow cells using velocity sedimentation in an isokinetic gradient ficoll in tissue culture medium.

A technique for the purification of rat megakaryocytes is described. Velocity sedimentation in a previously described isokinetic gradient of Ficoll (polysucrose) in tissue culture medium was more effective than isopycnic sedimentation for the purification of megakaryocytes and resulted in preparations of magekaryocytes which contained 2.4 +/- 0.8% (range 1.85-3.60%) megakaryocytes. Megakaryocytes exhibited a broad range of density between 1.06 and 1.15 gm/ml. The inaccuracy which is inherent in the use of velocity sedimentation without isopycnic sedimentation as a means of particle size analysis is discussed.


Analysis of the chemotactic response during aggregation in Dictyostelium minutum.

Measurements on the chemotactic aggregation of the amoebae of the cellular slime mould Dictyostelium minutum are reported. With the aid of previous theoretical results, the observed patterns of early aggregation on 2- and 3-dimensional substrates are interpreted. There is no statistically significant difference in the patterns in the 2 cases. The results suggest that the simplest plausible chemotactic law for this system is the relative gradient law in which the response velocity of the amoebae is proportional to the relative acrasin gradient. The results also suggest that the differentiation of the founder cell, which initiates aggregation, occurs before all cells have acquired chemotactic sensitivity. A diffusible chemical may be involved in the development of the latter.


Stationary phase and the cell cycle of Dictyostelium discoideum in liquid nutrient medium.

Cells of the axenic strain of the cellular slime mould Dictyostelium discoideum, AX-3, multiply in the liquid nutrient medium HL-5 with a doubling time of 12 h. When the cell concentration reaches approximately 1 X10(7) per ml the rate of cell multiplication begins decreasing and after 20-30 h reaches zero, at a stationary phase cell concentration of 2 to 2-5 X 10(7) cells per ml. The intercept of the extrapolated log phase and stationary phase plots has arbitrarily been considered the onset of the stationary phase. We have found that after cells have been in stationary phase for 24-32 h, mean cell volume increases by 25%, average dry weight by 37%, and average protein content by 24%. These values are close to the expected values for a cell population which is blocked at a point late in the cell cycle. Stationary phase cells also contain 25% more nuclear DNA than log phase cells, indicating that the population of cells at stationary phase is blocked after the DNA replication phase. Finally, when stationary phase cells are washed free of stationary phase medium and reinoculated into fresh medium, they reinitiate cell division synchronously. In the light of the demonstrated relationship between stationary phase and the cell cycle, a possible role for the growth inhibitor produced at stationary phase is considered.


Ultracryotomy of biological tissues to preserve membrane structure.

A vacuum transfer stage is described which permits visualization of ultracryotome sections without the considerable distortions found in sections which have either thawed or rehydrated after freeze drying. Membrane structure-nucleus, nucleolus, mitochondria with their cristae and plasma membrane-was observed only in cells at the surface of the tissue which had undergone the fastest freezing rates. Inspection of knife damage in sections through these superficial cells showed that glass or diamond knives which are perfectly adequate for sectioning resin-embedded tissues are less suited to sectioning frozen biological tissue. Deeper in the tissue, where the freezing rates were slower, ice crystal cavities destroyed all membranous structures.


Titration of Newcastle disease virus and its neutralizing antibodies in microplates by a modified hemadsorption and hemadsorption inhibition method.

Using the microtiter system, titration of Newcastle disease virus infectivity and neutralizing antibodies was carried out in chicken embryo fibroblasts grown in "U" or flat-bottomed plates. Infectivity was detected by a combined hemadsorption-hemagglutination method. Inhibition of that reaction indicated the presence of neutralizing antibodies. A 24-h microneutralization test was developed and compared to the plaque neutralization and microhemagglutination inhibition test. Reproducibility of the microneutralization test was statistically analyzed.


Effects of testosterone propionate administered perinatally on sexual behavior of female ferrets.

Female ferrets that received injections of testosterone propionate (TP) early in postnatal life displayed significantly more masculine behavior than did control females when gonadectomized and administered either TP or estradiol benzoate in adulthood. This increased masculine response potential was not correlated with the effects of early TP treatment on phallic development. In contrast to results obtained in most other species, perinatal administration of TP to female ferrets failed to disrupt their ability to display the behavior that is characteristic of the sexually receptive animal in estrus. When estrogenic stimulation was provided in adulthood, the receptive behavior of three groups of perinatally androgenized females was indistinguishable from that of both male and female controls. However, after gonadectomy and administration of TP, control males and females that had received TP prenatally plus on Day 3 were significantly more receptive than were control females. The induction of receptivity by TP was significantly inhibited by simultaneous administration of the antiestrogen MER-25.


Bovine pancreatic lipase. III. Lipolysis of oils and fats and fatty acid specificity.

Purified bovine pancreatic lipase hydrolyzed butteroil, vegetable oils, and synthetic glycerides. The enzyme hydrolyzed triglycerides more rapidly than di- and monoglycerides and tripropionin faster than any other synthetic glyceride. Triacetin was the least hydrolyzed glyceride. Gas-liquid chromatographic analysis of the free fatty acids liberated by the lipase from milk fat indicated that the enzyme selectively liberated butyric acid in higher proportion than the relative amount originally in the fat. The enzyme released saturated as well as unsaturated fatty acids from commericial vegetable oils. With regard to the lipolytic behavior, in general, the bovine pancreatic lipase closely resembled milk lipase.


Quantification of polyacrylamide gel electrophoresis for analysis of whey proteins.

Polyacrylamide gel electrophoresis of whey proteins has been quantified by standardization of the separation and staining procedure. During each electrophoresis experiment, a standard solution of whey proteins was separated and stained under the same conditions as the test material. In this way, proteins in the standard solution were subjected to identical processing conditions as the test samples. Densitometric scanning of the stained protein-containing gels followed by peak area determinations was carried out. By comparison with the standard peak areas, individual protein concentrations of the test samples were determined.


Parish clergy and the aged: examining stereotypes.

Hypotheses concerning clergymen's enjoyment of their pastoral contacts with older parishioners are examined using data from a national probability sample of 654 American Baptist parish ministers. The hypotheses test the ideas that ministers do not enjoy pastoral contacts with the elderly and that these contacts reflect the clergymen's ageist preferences and concern with instrumental over expressive values. The findings suggest that: (1) the majority of the clergy studied do not seem to have an aversion to ministering to older people; while not among the most enjoyable of their activities "ministering to the aged" clearly is not among the least enjoyable either; (2) there is some support for the view that clergy respond to the elderly in an ageist manner; comparing activities involving different age groups, the clergy prefer the young and adults to the aged; (3) those clergy who share with the elderly an interest in expressive activities are more likely to enjoy ministering to the aged.


Relationships between hearing loss and cognition in normally hearing aged persons.

The relationships between mild hearing losses and cognitive functioning were studied for two independently selected samples of aged subjects whose hearing was within normal limits. One group had 47 males of mean age 71.5 (SD 4.8) who were selected for their excellent health status. The second group consisted of 38 females of mean age 75.9 (SD 5.3), all of whom had some significant physical pathology. Hearing losses at various frequency levels (from 125 to 8000 cps) were correlated with performance on cognitive tests such as the WAIS, with age effects then being partialled out. The results reveal substantial associations between hearing losses and scores achieved on the intellectual measures for both samples. Verbal type tests show these relationships much more extensively than the performance tests. The findings imply that aged subjects may be more intellectually capable than their test performances suggest and that hearing is an important variable to be considered in the assessment of their cognitive functioning.


The effect of corticosteroids on human IgG synthesis.

The effect of prolonged, high-dose corticosteroid therapy on total IgG synthesis rates by human bone marrow and splenic leukocytes was studied in patients with immune thrombocytopenia. Marrow IgG production rates begin to decrease 3 weeks after beginning therapy and reach levels approximately one-fourth of pre-treatment rates after 6 weeks. No effect of corticosteroids on splenic IgG production rates could be shown. Marrow IgG synthesis rates in these patients were on the average from 3 to 10 times greater than corresponding splenic production rates and, in addition, appeared to correlate with serum IgG levels. These data suggest that the marrow is an important contributor to the total body IgG pool; and since corticosteroids appear to suppress marrow IgG production, this may be one reason for their therapeutic usefulness in antibody-mediated diseases.


Influence of different antigen doses on number and properties of antigen binding cells.

The number of antigen binding cells, as revealed by capping experiments, is strongly influenced by the concentration of antigen used. At high concentrations of antigen an additional population of cells of the size of lymphocytes can be detected, which is not stained after incubation with low doses of antigen. These cells bear immunoglobulin on their surface and can be removed from the spleen by carbonyl iron treatment.


A comparison of the effects of neonatally administered testosterone, testosterone propionate and dihydrotestosterone on aggressive and sexual behaviour in the female golden hamster.

Female golden hamsters received one of the following treatments on the day following birth: (i) 300 mug testosterone propionate in arachis oil, (ii) 300 mug testosterone in oil, (iii) 300 mug dihydrotestosterone in oil, or (iv) oil alone. As adults all animals, underwent three tests for behaviour. First, while intact, females were observed in aggressive interactions with males. Secondly, after ovariectomy, females were primed with oestrogen + progesterone and tested for receptivity with a stud male. Thirdly, all ovariectomized females were primed with testosterone propionate and tested for male patterns of behaviour with a receptive female. Compared with the effects of oil administration (control), testosterone propionate administration resulted in increased aggressiveness and the capacity to show male patterns of sexual behaviour, together with a decreased capacity to show female patterns of sexual behaviour. Testosterone increased aggression and male sexual behaviour, but had no effects on receptivity, while dihydrotestosterone decreased some components of receptivity but had no effects on aggressiveness or the capacity to show male mating behaviour.


RNA and protein metabolism in the oviduct and endometrium of the ewe at pro-oestrus: Regulation by oestradiol and progesterone.

The effects on RNA and protein metabolism in the oviduct and endometrium at pro-oestrus of oestradiol and progesterone secreted during the oestrous cycle were examined, using the ovariectomized, hormone-treated ewe as a model system. Thirty ewes received hormone injections during a period of 13 days, according to schedules designed to simulate endogenous ovarian secretion of oestradiol and progesterone during the oestrous cycle. Hormone effects on RNA:DNA ratios and on rates of synthesis of protein and methylated RNA in vitro, as well as effects on oviducal and uterine weight, were examined. The results obtained suggest that endogenous ovarian hormones have the following effects in the intact ewe. The secretion of oestradiol at pro-oestrus rapidly increases rates of synthesis of protein and methylated RNA, and mean cell content of RNA in both the endometrium and oviduct. Oestradiol secreted during the previous luteal phase of the oestrous cycle markedly increases mean cell content of RNA and amounts of protein and methylated RNA synthesis occurring in both tissues at pro-oestrus. In the endometrium, progesterone secreted during the luteal phase increases the RNA:DNA ratio, and probably also the amounts of protein and methylated RNA synthesized at pro-oestrus, but there are no significant interactions between the effects of oestradiol and progesterone. Progesterone had no effect on either the amounts or rates of synthesis of protein or methylated RNA in the oviduct. The results are discussed in relation to the hormonal regulation of physiological functions of the oviduct and endometrium during the first few days after the onset of oestrus.


Effects of pinealectomy on the rat oestrous cycle and pituitary gonadotrophin release.

In 4-day cyclic rats kept in a room with the lights on from 05.00 to 19.00 h, sham pinealectomy or pinealectomy on the morning of pro-oestrus did not alter the length of the oestrous cycle for 44 days or the time and magnitude of the rises in LH, FSH and prolactin in the circulation in the afternoon on pro-oestrous days 0, 20 or 44. On day 45, the light schedule was set forward 4 h to run from 09.00 to 23.00 h. The rats continued to have seven additional consecutive 4-day oestrous cycles. On day 27 after the resetting of the light schedule, the pro-oestrous rises in serum LH, FSH and prolactin were delayed 4 h in all rats and a normal quota of eggs was ovulated that night. Other 4- and 5-day cyclic rats which had been made persistently oestrous by anterior deafferentation of the medial basal hypothalamus (AC) underwent pinealectomy. These AC-pinealectomized rats were ovariectomized 60 days later and histological examination of the ovaries revealed no evidence of recent ovulation. Five to six weeks after ovariectomy, sequential blood samples were withdrawn through indwelling atrial cannulas in the AC-pinealectomized-ovariectomized rats and in ovariectomized, pinealectomized-ovariectomized and AC-ovariectomized rats. Regular pulsatile rhythms in plasma LH were measured in all rats. Subcutaneous injection of 50 mug mug oestradiol benzoate in oil lowered plasma LH levels in all four groups but caused an LH surge in the afternoon 2 days later only in the ovariectomized and pinealectomized-ovariectomized rats. The results indicate that the pineal gland in rats kept on a 14 h light: 10 h darkness schedule does not play an active or premissive role in the timing or magnitude of LH, FSH or prolactin release at pro-oestrus, the length of the oestrous cycle, or LH release in ovariectomized rats.