Quantitative observations on iliac bone marrow mast cells in chronic renal failure. Mast cells have been counted in sections of iliac bone from 61 control subjects at necropsy. Mast cells were found in all but three, and the range was 0-33-7, median 1-95 per mm2 marrow. The majority (82%) had less than 4-99 mast cells per mm2 marrow; in 37-7% there was less than 1 mast cell per mm2 marrow. In a group of 45 patients with chronic renal failure there was a significant increase in the numbers of mast cells (P less than 0-001) with a range of 0-96-55-63, median 9-55 per mm2 marrow. Mast cells were common in the areas of marrow fibrosis associated with osteitis fibrosa but this was not the sole cause of the increase since there was also an excess of mast cells in the non-fibrous parts of the marrow. There was a tendency towards greater numbers of mast cells in those cases with most marked osteitis fibrosa in association with the prominent marrow fibrosis, but there was no significant relationship between mast cell numbers and other features of oesteitis fibrosa such as the number of osteoclasts and the amount of woven bone formation. There was no relationship between the numbers of mast cells and the amounts of total bone, ostoid, percentage mineralization of cancellous bone, or the presence of osteomalacia.


Agar culture and chromosome analysis of eosinophilic leukaemia. The blood and bone marrow of a 75-year-old woman with eosinophilia were studied in an attempt to characterize further the entity eosinophilic leukaemia (EL). Haemopoietic cell culture in agar gave patterns of colony size and number and cell morphology indistinguishable from those seen in classical chronic granulocytic leukaemia (CGL). Analysis of myeloid cells showed an extra C group chromosome that proved on Giemsa banding to be number 10. These results suggest that some cases of EL represent a chronic myeloproliferative disease that resembles classical CGL but can be distinguished from it by chromosomal analysis.


Change of blood group from A2 to Ax in a child with congenital abnormalities. A blood group change from A2 to Ax is described in a boy who had Fallot's tetralogy, oesophageal atresia, and a tracheo-oesophageal fistula. The change of blood group created a transfusion problem. The unusual serology is discussed.


Detection and quantitation of tetanus antitoxin in blood donations. Passive haemagglutination and IEOP have been used both to detect and to measure tetanus antitoxin in human donor sera. Forty percent of blood donors had detectable antitoxin but only 9% had levels suitable for production of human antitetanus immuoglobulin (larger than or equal to 2 IU/ml). The incidence of high titre antitoxin was significantly greater in men and was unrelated to the ABO blood group system. The prevalence of antitoxin in selected donor groups and immunized staff is shown.


Factor VIII levels during the course of acute hepatitis in a haemophiliac. A 51-year-old patient with haemophilia since childhood (usual factor VIII level 14%) developed acute viral hepatitis type B two months after an operation which had been covered by cryoprecipitate. The course of the hepatitis following admission was severe with encephalopathy and ascites. Evidence of intravascular coagulation with an increased radioactive fibrinogen turnover was also present. The factor VIII level measured by a one-stage clotting factor assay rose rapidly to 200% of normal and remained at this level for two weeks, and factor-VIII-related antigen as measured by electroimmunoassay also became greatly elevated (900% of normal). The possible mechanisms underlying those surprising changes are discussed.


Heparin neutralizing activity test in the diagnosis of acute myocardial infarction. We have previously shown in patients after recovery from a myocardial infarct (post-MI) that the heparin neutralizing activity (HNA) in the plasma is raised and that this activity may come from platelet factor 4 derived from activated platelets. This report concerns 89 patients admitted with acute chest pain; in 54, with evidence of acute infarction, the level of HNA is much higher than in a post-MI group or controls. Over the ensuing weeks the HNA decreases to the post-MI level. In 34 patients the evidence subsequently collected excluded a diagnosis of infarction; in these there were almost always normal amounts of HNA and little overlap with the results from the patients with infarcts. This easy test is therefore likely to prove clinically useful. Its significance is discussed. The platelet count and platelet volume are both abnormal in patients with acute infarction and also in the chest pain group so these tests do not help to discriminate.


Comparison of tablets and paper discs for antibiotic sensitivity testing. The value of tablets and paper discs as reservoirs of antimicrobial agents for use in sensitivity testing was compared. Antibiotics that were unstable in paper discs showed no demonstrable loss of activity in tablets over a period of 50 days under adverse storage conditions. The antibiotic content of commercially prepared tablets is very high in comparison with the accepted content of paper discs used in Britain, but not all of the agent is released from tablets during tests. Comparison of the size of zones of inhibition around tablets and standard paper discs indicated that the amount of the various agents released from the tablets varied between 2-6% and 69% of the stated content. In tests of the sensitivity of a range of common pathogenic organisms, the results obtained with the tablet method--when interpreted as recommended by the manufacturer--were generally similar to those obtained with a paper disc method commonly used in British laboratories. In 47% of tests with aminoglycoside antibiotics, however, strains sensitive by the disc method were 'intermediate' or resistant by the tablet method. As with paper discs, it was necessary to press the tablets on to the medium. With adjustment of the 'effective antibiotic content of tablets to bring it into line with the accepted content in paper discs, the stability of antibiotics in the tablets might make them an acceptable alternative to paper discs.


Pseudomonas putrefaciens as a cause of infection in humans. Pseudomonas putrefaciens, a strongly H2S-producing pseudomonad, was isolated from 10 human infections over a two-year period. In one patient the organism was repeatedly isolated from a phlegmone developing in the depth of a varicose leg ulcer. This is the first report on the occurrence of Ps. putrefaciens in humans outside the USA and the first to provide the detailed account of a clinical observation where the opportunistic pathogenic role of this unfamiliar organism has been sufficiently documented. Data are presented on the bacteriological properties and on the antibiotic sensitivity of Ps. putrefaciens.


Olfactory relationships of the telencephalon and diencephalon in the rabbit. II. An autoradiographic and horseradish peroxidase study of the efferent connections of the anterior olfactory nucleus. The efferent connections of the anterior olfactory nucleus in the female albino rabbit have been studied using the autoradiographic and horseradish peroxidase methods for tracing axonal pathways. Following a unilateral injection of 3H-leucine into the olfactory peduncle, radioactively labeled efferent projections from the anterior olfactory nucleus were traced into all layers of the ipsilateral main olfactory bulb beneath the olfactory nerve layer and through the ipsilateral anterior limb of the anterior commissure and plexiform layer of the medial side of the cerebral hemisphere to the deep half of the plexiform (IB) and pyramidal cell (II) layers of the prepyriform cortex, the tenia tecta, and the entire surface of the olfactory tubercle. Labeled projections crossing the midline within the anterior commissure were followed to the layers IB and II of the contralateral anterior prepyriform cortex and pars externa, pars lateralis, and pars dorsalis of the anterior olfactory nucleus, and through the periventricular layer of the olfactory peduncle to all layers of the main olfactory bulb beneath the olfactory nerve layer. No well-defined labeled projection was traced to the contralateral accessory olfactory bulb. Evidence for possible anterior olfactory nucleus and/or prepyriform cortical projections to the ipsilateral paleocortical half of the claustrum, horizontal limb of the nucleus of the diagonal band, the posterior lateral hypothalamus at the level of the mammillary complex, and to the bed nucleus of the stria terminalis is discussed. Intra-axonal retrograde transport of horseradish peroxidase from axon terminals to parent cell bodies after unilateral injection of the protein into the main olfactory bulb or anterior olfactory nucleus revealed that anterior olfactory nucleus projections to the olfactory bulbs and the contralateral anterior olfactory nucleus arise predominately from the pars externa. The autoradiographic data indicate that the anterior olfactory nucleus projects to olfactory cortical structures which also receive afferent input from the olfactory bulb and that the termination of these projections is complementary to those from the olfactory bulb.


Organization and development of brain stem auditory nuclei of the chicken: tonotopic organization of n. magnocellularis and n. laminaris. Extracellular recordings of responses to tone-burst stimulation were used to determine the tonotopic organization of n. magnocellularis (NM) and n. laminaris (NL) in hatching chickens. NM cells show "primary-like" response patterns to ipsilateral stimulation, and are arranged in dorso-ventral isofrequency columns. Units responding to the highest frequency tones (about 4,100 Hz) are situated at the rostromedial pole of the medial division. Units with lower characteristic frequencies (CF's) are found at successively caudal and lateral sites, until extremely low CF's ( less than 500 Hz) are represented dorsoventrally in the daudolateral tail of the lateral division. No evidence was found of auditory input to the region which receives projections from the macula lagena. NL receives polarized, binaural, excitatory input. Units have similar CF's and thresholds to tones presented to either ear. The tonotopic organization in NL matches that found in NM--high CF's rostromedially and low CF's caudal and lateral. Quantitative procedures were developed for relating CF to the position of a unit within either nucleus. These analyses account for 79% and 89% of the frequency variance found within NM and NL, respectively, and predict the CF of a neuron by its position within each nucleus.


Organization and development of brain stem auditory nuclei of the chicken: organization of projections from n. magnocellularis to n. laminaris. The tonotopic and topographic organization of the bilateral projection from second-order auditory neurons of nucleus magnocellularis (NM) to nucleus laminaris (NL) was examined in young chickens. In one group of birds, the NM axons which innvervate the contralateral NL were severed by cutting the crossed dorsal cochlear tract at the midline. Heavy terminal degeneration in NL was confined to the neuropil area immediately ventral to the perikaryl lamina. Very little degeneration was seen in the dorsal neuropil region. In a second series of animals, the charactertistic frequency (CF) of cells in an area of NM was first determined by microelectrode recording techniques and then a small electrolytic lesion was made through the recording electrode. Following survival periods of 24-48 hours, the distribution of projections from the lesioned area to the ipsilateral and contralateral NL was examined using the Fink-Heimer method. As previously described in the pigeon, projections from NM terminate densely in the neuropil region immediately dorsal to the ipsilateral NL cell bodies and ventral to the perikaryl layer on the contralateral side, providing each NL neuron with segregated binaural innervation. Lesions in any area of the NM produced degeneration confined to a limited caudo-rostral and medio-lateral portion of both laminar nuclei. To investigate this topographic relationship, the cuado-rostral extents of the lesion in NM and of the resulting degeneration in both NL were determined. Linear regression and correlation analyses then related these positional values to each other and to the CF found at the center of each lesion. All correlations were highly significant and ranged from 0.78 between the position of the lesion in NM and CF to 0.91 between the caudo-rostral position of degeneration in the NL ipsilateral and contralateral to the lesion. It is concluded that neurons in NM project in a very discrete topographic, tonotopic and symmetrical fashion to NL on both sides of the brain, contributing to the binaural response properties and tonotopic organization of neurons in NL. The results also suggest that the organization of projections from NM to NL could provide a mechanism for the differential transmission delay required by a "place" model of low-frequency sound localization.


The olivocerebellar projection in the cat studied with the method of retrograde axonal transport of horseradish peroxidase. The distribution of labeled cells in the inferior olive of the cat has been mapped following injections of small amounts of horseradish perosidase in the paramedian lobule of the cerebellum. The distribution of labeled cells was plotted in drawings of approximately serial transverse sections. The findings in each case were transferred to a standard diagram of the olive to facilitate comparison of cases. Previous studies of the distribution of retrograde cell loss in the inferior olive following cerebellar lesions (Brodal, '40b) showed that fibers ending in the paramedian lobule come from the caudal part of the ventral lamella of the principla olive. This was confirmed with the peroxidase method, but in addition three other separate and well circumscribed area of the olive showed labeling: one in the dorsal accessory olive, another in the rostral part of the medial accessory olive, a third in the caudal part of the dorsal lamella of the principal olive (fig. 7). There is some degree of topical arrangement within the projection of each of these olivary areas to the paramedian lobule. It is particularly striking that the projection areas of the caudal one-third of the lobule are different from and overlap only little with those of the orstral two-thirds. On account of diffusion of the injected perosidase solution in the folia it could not be decided whether the different olivary areas project to particular longitudinal zones in the paramedian lobule. The main findings can be correlated with the physiological observations of Armstrong et al. ('74). Some of the "paramedian" olivary areas are labeled also following peroxidase injections in other cerebellar parts, among them the nuclei interpositus anterior and posterior. The findings are compatible with the notion that olivocerebellar fibers branch to supply more than one cerebellar region. It is confirmed that the olivocerebellar projection, including that of the nuclei, is almost completely crossed. In the discussion it is emphasized that afferents from several sources converge on all four olivary regions projecting onto the paramedian lobule. The olivocerebellar projection obviously allows for divergence as well as convergence of impulses from the olive to the cerebellum. For further insight into the anatomical organization of the inferior olive, the entire olivocerebellar projection has to be mapped with the peroxidase methods, and further studies of the afferents to the olive are needed. In such studies, as well as in physiological ones, it is essential that findings are described with meticulous reference to the topography of the olivary subdivisions.


A normally laminated afferent projection to an abnormally laminated cortex: some olfactory connections in the reeler mouse. The relative positions of pyramidal and polymorphic cell classes are inverted in the central olfactory cortical structures of the reeler mutant mouse. Each cell class is generated at the normal embryonic time. The polymorphic cells of the mutant, like those of the normal, are generated between E11-E13. The pyramidal cells are formed between E11-E16 in both. Despite the anomalous positions of their somata deep in the cortex the apical dendrites of many pyramidal cells reach and ramify at a superficial cortical level subjacent to the lateral olfactory tract. The main and accessory olfactory bulbs are cytoarchitectonically normal in the mutant and project normally upon the anterior olfactory nucleus, the olfactory tubercle, the hippocampal rudiment, the piriform cortex, the amygdaloid region and the entorhinal cortex. As in the normal animal the axons transverse layer Ialpha, and their terminals are concentrated in the immediately subjacent laminar zone. The rostrally directed cortic-cortical association system of the piriform cortex projects upon the anterior olfactory nucleus in the mutant just as in the normal with a relative concentration of terminals in a lamina subjacent and complementary to the zone of termination afferent systems in the abnormally laminated olfactory cortex of the mutant syggests that, in this system at least, the developmental mechanisms which determine relative position of neuron somata and those which govern axon trajectories and the distribution of axon terminals are largely independent.


Anatomy of the cerebral ganglion of the male acanthocephalan, Moniliformis dubius. An atlas of the cerebral ganglion of Moniliformis dubius has been constructed and the cells identified by number. There is a total of 88 cells, two of which are binucleate. These cells (20 and 90) are located in the frontal plane. The cell population is equally distributed between each half of the ganglion, there being 41 cells on each side with six cells centrally located between the two halves. Most cells give rise to bilateral processes and are organized in such a way that there is an outer coat of somata or cell bodies. Cell bodies predominate on the ventral surface, but the dorsal surface is more of a mixture of cell bodies and nerve processes. The core or neuropile occupies the central part of the ganglion and is the site for cross over for many cell processes which then exist from the side opposite the cell body. The cytoarchitecture of these cells varies considerably. Most have a large round nucleus with well defined nucleoli.


Functional localization and cortical architecture in the nine-banded armadilli (Dasypus novemcinctus mexicanus). A functional map of the armadillo neocortex was produced by cortical stimulation and recording evoked potentials following somatic, auditory and visual stimuli. The results obtained were then correlated with the cortical architecture as revealed by Nissl, Golgi and myelin-stained sections. Cortex rostral to the supraorbital sulcus has a wide layer IV and is mostly silent, except for a motor eye field and a part of the tongue sensory region in its caudal part. Two types of motor-sensory cortex are present caudal to the supraorbital sulcus. Postsupraorbital I is mostly motor and has prominent pyramidal layers. Layer V is particularly well developed and in rostral sections its superficial zone is broken up into clusters similar to the solid "barrels" seen in layer IV of other species. Postsupraorbital II has less prominent pyramidal layers and layers II and III are organized into clusters. This region corresponds to the sensory area for the limbs and trunk and the partially overlapping (surface recordings) sensory and motor areas for head, snout and tongue. Digits and limbs are rostral to the trunk representation in both the sensory and motor "homunculi." Even though surface recording was employed, potentials evoked by visual stimuli could only be recorded from a small caudal area with a very thin layer IV. Although striate and peristriate areas appear similar in Nissl stained preparations, they can be readily differentiated in Weil stained sections. The stellate character of neurons in layer IV of the visual cortex is particularly apparent in Golgi material. Auditory evoked surface potentials were recorded from a broad oval region in the caudal lateral cortex which has a wide layer IV and aggregates of neurons in layers II and III. A Weil stain demonstrates inner and outer bands of Baillarger in this same region. The presumptive insular cortex is electrically silent to sensory stimulation and presents as a narrow band just dorsal to the rhinal fissure with indefinite cell lamination and little myelin.


Nonequilibration of acetyl units from fatty acids and glucose in mammary glands of lactating mice. Lipid synthesis by lactating mouse mammary gland slices from [1-carbon14] propionate, [1-carbon14] valerate, and [1-carbon-14] heptanoate, in the presence of glucose and from [uniformly labeled carbon-14] glucose in the presence of propionate, valerate, and heptanoate was studied. Regardless of the labeled substrate, almost all of the radioactivity incorporated into the total lipid fraction was in triglycerides. In experiments with labeled glucose and either the propionic. the valeric, or the heptylic acid, the carbon-14 fatty acids in triglycerides were almost exclusively composed of even-chain fatty acids. When the substrate was either [1-carbon-14] propionate, [1-carbon-14] valerate, or [1-carbon-14] heptanoate, in the presence of glucose, the pattern changed to one in which a larger proportion of the radioactivity was in the odd-chain fatty acids. These findings suggest that in mouse mammary gland, acetyl units produced via beta-oxidation of fatty acids may not equilibrate with those formed from oxidation of pyruvate which is derived from glucose.


Milk secretory response of the conscious lactating rat following intravenous injections of rat prolactin. Mammary glands of rats on day 14 of lactation were emptied of milk by the pups after 8 h of nonsuckling with the aid of oxytocin injections to the mother. The glands subsequently refilled about 50% within 6 h and were completely refilled with mild within 16 h. Suckling for 5 min midway during the 8 h period of nonsuckling caused complete refilling of the glands within 6 h. Refilling within 6 h also occurred if, instead of suckling, rat prolactin was injected at 1 min intervals directly into the circulation of the conscious mother via an indwelling right atrial catheter. Five injections of 200 ng or three injections of 400 ng were without effect (in comparison with five injections of saline) upon mammary refilling whereas 10 injections of 200 ng or four injections of 400 ng simulated significant but submaximal refilling. Complete refilling resulted from five injections of 400 ng. Prolactin is secreted steadily over prolonged periods of suckling in the rat, but only that prolactin released during the first few minutes of suckling appears to be required for milk secretion.


Metabolism of arginine and ornithine in the cow and rabbit mammary tissue. Amino acid uptake by the bovine mammary gland was determined by arteriovenous difference. Extraction of arginine from the plasma by the lactating bovine mammary gland was in excess of requirements for milk protein synthesis. Ornithine and citrulline also were extracted by the gland but are not in milk protein. Incubations of slices of lactating mammary tissue from cows and rabbits indicate that nonessential amino acids, especially proline and glutamate, are the major end products of arginine and ornithine metabolism in the lactating mammary gland.


Effect of drying off practices on mastitis infection. Two methods of drying off cows, intermittent milking and abrupt cessation, were studied with data from 9254 quarters of cows on 36 New York dairy farms. Eighty percent of the cows were infused with nine different antibiotic preparations separately at drying off, and 20% served as controls. Cows dried off by intermittent milking has a similar number of quarters infected at drying off, had fewer spontaneous recoveries, had a higher rate of cure, and developed fewer new infections in control quarters in comparison with cows dried off by the stop method. Methods worked equally well in treated cows. However, intermittent milking resulted in fewer infections at subsequent calving than stop milking in nondry treated cows. Cows producing less than or equal to 4 kg of milk at drying off were more highly infected than higher producing cows. Hind quarters contained more infections at drying off, fewer responded to therapy, and more infections developed in the dry period. Cows with dry periods of less than or equal to 30 days had more infected quarters respond to therapy and had the fewest new infections in the dry period. The role of routine dry cow therapy in decreasing the number of infections in dairy herds by preventing new infections and removing old infections is demonstrated.


Metabolism of liquid whey fed to sheep.

Metabolism of liquid whey fed to sheep. Digestion was studied with three groups of four sheep each fed alfalfa hay. Treatments were whey only, whey plus water, and water only. Digestibilities of dry matter for the three rations were 71.3, 69.7, and 62.9%. Digestibilities of whey solids calculated by difference was 86.9% for whey only and 81.6% for whey plus water. Twelve sheep were in three trials to determine the intake of whey, water, and hay consumption. They were assigned to whey only, whey plus water, and water only. Hay, whey, and water consumption was 1.89, 8.73, and 0; 1.88, 7.62, and 1.21; and 2.05, 0, and 6.07 kg per animal daily. Three mature fistulated wether sheep were fed 6 liters of liquid whey daily. Three mature fistulated wether sheep uere fed 6 liters of liquid whey daily, and three similar sheep were fed water only. Rumen liquor samples were taken for analysis of volatile fatty acids at 0, 3, and 6 h after whey feeding on each of three alternate days. Alfalfa hay (1.2 kg/animal daily) was fed after the 6-h sampling. Molar percentage of acetic and valeric acids were lower and for butyric acid was higher for sheep fed whey. Propionic acid, acetate:propionate ratio, and total concentration were not affected significantly by whey feeding. Acetic acid decreased from 0 to 3 h, then partly returned to the prefeeding level at 6 h. The reverse was true for butyric acid. Valeric acid and total concentration of volatile fatty acids decreased from 0 to 6 h. Effects of day were unexplained.


Digestibility and volatile fatty acids in steers and wethers at 21 and 32 C ambient temperature. Four steers and four closely shorn wethers, each with ruminal fistula, were used in a reversal experiment to compare the direct effect of 21 and 32 C ambient temperatures (70% relative humidity) on dry matter digestibility and ruminal volatile fatty acid concentrations. Pelleted alfalfa intake was held constant at 2.3% of body weight. Orts were placed into the rumen via fistula. Dry matter digestibility was determined by total collection during the last 7 days of each trial. On one of the last 2 days, ruminal fluid was sampled 11 times within 13 h. At 32 C, the steers suffered considerable heat stress as evidenced by a mean 1.6 C increase in body temperature, reduced heart rate, and greatly increased respiration rate. The wethers, however, exhibited only increased respiration rate at 32 C. Extent of digestion increased five percentage units for the steers but did not change for the wethers at the high temperature. Temperature caused only slight shifts in ruminal volatile fatty acid concentrations. Volatile fatty acid concentrations were much higher in steers than in wethers. The frequently observed effects of heat stress on intake and digestibility are mediated by changes in rate of passage as a result of altered thyroid secretion rates.


Relationship between ruminal ammonia and nonprotein nitrogen utilization by ruminants. I. Development of a model for predicting nonprotein nitrogen utilization by cattle. The influence of ration composition on mean ruminal ammonia concentration was studied by collecting samples of ruminal ingesta from cattle fed rations varying in crude protein and total digestible nutrient content. A minimum of four sampling times distributed throughout the day permitted calculation of mean ruminal ammonia concentrations. Mean ruminal ammonia concentration was positively related to dietary crude protein concentration and negatively related to total digestible nutrient concentration. It is postulated that mean ruminal ammonia concentration may be a useful criterion for predicting efficacy of nonprotein nitrogen supplementation. A quantitative approach for evaluating nonprotein nitrogen supplementation based upon determination of the point at which ruminal ammonia exceeds the requirement (5 mg ammonia nitrogen/100 ml rumen fluid) of the ammonia-utilizing bacteria is proposed. Dietary conditions expected to result in excessive concentrations of ruminal ammonia are defined and recommended upper limits for nonprotein nitrogen supplementation are presented. Theoretical relationships between composition of the unsupplemented ration, amount of nonprotein nitrogen added, and efficiency of nonprotein nitrogen utilization are discussed. The practice of adding nonprotein nitrogen so as to exceed 12 to 13% crude protein in typical dairy or feedlot rations needs to be reevaluated.


Relationship between ruminal ammonia and nonprotein nitrogen utilization by ruminants. II. Application of published evidence to the development of theoretical model for predicting nonprotein nitrogen utilization. Results from published experiments dealing with several aspects of nitrogen utilization by ruminants were used to test the concept of zero utilization of nonprotein nitrogen under conditions where more ammonia (greater than 5 mg ammonia nitrogen/100 ml) is in the rumen than can be converted to microbial protein. Results from experiments where the flow of non-ammonia nitrogen to the abomasum of sheep was measured indicate that when urea was the source of supplemental nitrogen, a constant quantity of amino acids reached the abomasum for all rations ranging from 10 to 23% crude protein. From growth studies, addition of nonprotein nitrogen to low protein, high energy rations caused an improved rate of gain. Additions of nonprotein nitrogen to rations resulting in predicted ruminal ammonia concentrations greater than 5 mg ammonia nitrogen/100 ml rumen fluid were without benefit. From lactation studies, nonprotein nitrogen supplementation did not improve milk production if the ration contained more than 12.5% crude protein prior to supplementation or if the predicted ruminal ammonia concentration was greater than 4 mg ammonia nitrogen/100 ml rumen fluid. The importance of the amino acid requirement of the animal as well as the composition of the ration in designing and evaluating nitrogen supplementation studies is discussed.


Simplified method for producing permanent ruminal fistulae. A quick and simple method for establishing permanent rumimal fistulae is described. A frozen cannula is forced into the rumen in a manner similar to the use of a trocar. Minimal equipment is required, and the entire operation can be performed in 15 to 20 min in sheep or 30 min in cattle. The technique has been used successfully to fistulate seven sheep and two cows.


Effect of forage-to-concentrate ratio in complete feeds and feed intake on digestion of starch by dairy cows. In digestion trials, 16 Holstein cows were fed complete mixed feeds of (a) 75:25, (b) 60:40, (c) 45:55, and (d) 30:70 ratios of forage to concentrate on a dry basis. At an intake of feed for maintenance, digestibility of starch ranged from 96.2 to 96.8%. During the lactation phase when intakes of feed were 2.5 to 3.2 times maintenance, digestibility of starch ranged from 84.7 to 88.1%. Starch in feces represented 2.9, 3.7, 5.5, and 5.9% of the intake of dry matter during the lactation phase, and .6, 1.0, 1.3, and 1.6% of intake at maintenance for the respective rations. At high intakes of concentrate, substantial amounts of dietary starch escaped degradation in the rumen and small intestine and were lost in the feces.


Erythrocyte delta-aminolevulinic acid dehydrase changes in the preruminant calf. Sixteen preruminant, male Holstein calves were tested for erythrocyte delta-aminolevulinic acid dehydrase activity, hemoglobin and erythrocyte count in blood samples taken weekly during the first 13 wk of life. delta-Aminolevulinic acid dehydrase showed a continual increase in activity beginning at wk 3 to a maximal activity at wk 9. Hemoglobin content was decreased by wk 5 and erythrocyte count was reduced by wk 6. delta-Aminolevulinic acid dehydrase enzyme activities increased with age corresponding to changes in hemoglobin and erythrocyte count during early postnatal development of the preruminant calf.


Bronchial sensitivity to methacholine in current and former asthmatic and allergic rhinitis patients and control subjects. Ninety-eight subjects underwent methacholine (Mecholyl) aerosol challenge at a concentration of 5 mg/ml. All atopic groups differed significantly in their bronchial response to Mecholyl compared to nonatopic control subjects. Nineteen normal subjects had a mean decrease of 11.4% in forced expiratory volume in one second (FEV1) with 128 Mecholyl inhalations and 2 decreased by greater than 20%. Fifteen of 27 allergic rhinitis patients had a positive response to Mecholyl as defined by a decrease in FEV1 greater than 20%. Four of these responders developed greater diminution in FEV1 response with additional Mecholyl inhalations in a manner similar to that seen in the asthmatic patients. Eighteen current asthmatics showed the greatest bronchial sensitivity to Mecholyl with a mean 33% decrease in FEV1 at 4.5 inhalations. One hundred per cent of current asthmatics and 82% (28 of 34) of former asthmatics (free of asthma symptoms for 1 to 20 yr) showed positive response to Mecholyl. A 20% decrease in FEV1 with a total of 20 inhalations or less is consistent with a diagnosis of "current asthma," and even 20 of 34 former asthmatics responded in this manner. The Mecholyl aerosol test can be used to diagnose previous and current asthma and may be of value in predicting asthma in hay fever patients.


Stinging insect allergy: detection and clinical significance of venom IgE antibodies. Venom-specific IgE antibodies in 109 sera from patients who had had immediate systemic allergic reactions following insect stings were measured by the radioallergosorbent (RAST) procedure. The majority of sera contained IgE antibodies to either bee, yellow jacket, or hornet venoms. Some sera had positive RAST reactions with 2 or 3 venoms, but others contained single venom-specific IgE antibodies. Of 24 patients who had large local reactions, the sera of 12 contained venom IgE antibodies. The RAST procedure provides an accurate means of documenting IgE-mediated allergic sensitivity to stinging insects.


Delayed-type hypersensitivity reaction in the lungs of guinea pigs due to potassium dichromate. Potassium dichromate was inhaled by guinea pigs previously immunized by potassium dichromate until strong positive patch tests were obtained. No obvious respiratory changes were noted during and after inhalation. Histologically, however, mononuclear cells infiltrated the interstitial spaces in large areas of the lung, producing considerable thickening of the alveolar spaces in 24 to 48 hr after inhalation. Polymorphonuclear cells were predominant initially. These changes were similar to the delayed-type hypersensitivity reaction in the lung elicited by the inhalation of purified protein derivative (PPD) in the guinea pigs immunized by an injection of dry-killed tubercle bacilli. A less marked reaction was observed in guinea pigs passively sensitized with peritoneal exudate cells and lymph node cells. Consequently, the pulmonary changes were thought to be elicited by delayed-type hypersensitivity reaction due to a simple chemical. The clinical implications of delayed-type hypersensitivity reaction in the lung due to simple chemicals are discussed.


Operative and nonoperative risks in the cardiac patient. Four hundred and sixteen patients with documented arteriosclerotic heart disease (ASHD) underwent 424 diagnostic and therapeutic surgical procedures during the year 1970 at the Henry Ford Hospital. They were classified according to the specific clinical manifestation of their cardiac abnormality. Patients with a history of old, well-compensated myocardial infarction, and those with cardiac arrhythmia, bundle-branch block, congestive heart failure and A-V block (pacemaker-protected) but no evidence of previous myocardial infarction fared almost as well as subjects of the same age without cardiac disease, and were considered to run the lowest operative risk. Patients with angina, especially if there was a history of infarction, were an intermediate risk in terms of complications and mortality. Patients with a history of previous infarction complicated at the time of the surgical procedure by arrhythmia, A-V block, bundle-branch block, or congestive heart failure were in the "highest risk" category. A severe A-V block indicated the need for insertion of a "prophylactic" pacemaker before any attempt at a diagnostic or therapeutic procedure. No patient with clinical or electrocardiographic evidence of a recent infarction (less than three months' duration) should undergo any elective surgical procedure under any form of anesthesia unless the surgeon is prepared for a high mortality rate that may approach 90 percent. In contrast, the patient with old, well-compensated myocardial infarction and no evidence of dysrhythmia, block or congestive failure can tolerate even a major surgical operation under any form of anesthesia extremely well.


Membranous nephropathy: an overview.

Membranous nephropathy: an overview. Membranous nephropathy (MN) accounts for about 20 percent of cases of the nephrotic syndrome. The importance of renal biopsy in establishing the diagnosis is emphasized. In the great majority of MN patients, no etiologic factor can be discerned. In a significant minority, MN appears to be a manifestation of sarcoidosis, diabetes, lupus, syphilis, malaria, or toxicity from heavy metals or drugs. In some cases the "cause" is neoplasia (including lymphoma) or a viral infection. Massive proteinuria, hypoproteinemia and edema are the principal manifestations of MN, finally resulting in renal failure. Treatment consists chiefly of diet and diuretic drugs. In the more pronounced cases, corticosteroids may have a favorable effect and in very resistant cases, cyclophosphamide is indicated. Judicious use of these modalities if often associated with the diminution or disappearance of the clinical signs of MN.


[Contraception and subsequent fertility. General review on the reversibility of oral contraception]. We have carried out a general review of the problem of reversing the contraceptive mehtods using oestrogen-pregestogen contraceptives. We have very carefully reviewed the onset of long cycles showing clearly that they occur more easily in women who have started with irregular cycles, as do post-pill amenorrhoea cases. Spontaneous abortions seem to occur just as often after the pill as in the general population and the alarm that was raised by D. Carr seems to us to be very exaggerated. The infants are normal, have a lower perinatal mortality when we take into account the patients who use this method and there are no more congenital abnormalities than in other patients.


[Pseudosarcomatous leiomyomas of the uterus. Fusiform cell myomas, plexiform myomas and myomas with nuclear anomalies]. The authors report 14 cases of atypical myomata wich might have been mistaken for sarcomata were it not for the fact that they were devoid of mitotic features. Four of the myomata were immature types with small fusiform cells and without connective tissue fibres. Two of the myomata had a plexiform structure several cm. in diameter and appaered to correspond to tissue that was undergoing endothelial and periocytal regeneration round an old focus of necrosis. These features could only be made out precisely under the microscope. The other eight myomata were mature in form and associated with abnormal nuclei (large, hyperchromatic irregular multi-nuclear giant cell clumps) surrounding the periphery of centres of hypoxia, more commonly after pregnancy (3 cases) or treatment with progesterone (2 cases).


[[Utero-placental hemodynamics during normal and pathological pregnancies. Its changes under the influence of a beta-sympathicomimetic: salbutamol]. Utero-placental circulation was studied using a gamma-camera which measures the appearance of a radio-active bolus of transferin tagged with Indium 113 in the region of the placenta and of the myometrium. The technique adopted for the calculations that are carried out allows a ratio between the placental blood flow and the myometrial blood flow to be established. The measurements that were taken in the last trimester of pregnancy showed that there was a significant difference between the ratio in normal pregnancies and in those that resulted in the deliveries of small-for-date babies. The foetus is small-for-date in 80 per cent of cases when the placental blood flow (Dp) to the myometrial blood flow (Dm), ratio is less than 80. It is small-for-dates in 25 per cent of cases if the Dp/Dm is between 0.80 and 1.60. If the Dp/Dm ratio is about 1.60 the foetus is normal in 93 per cent of cases. When we compared especially the variations between the Dp/Dm ratio during the administration of Salbutamol, we showed in the light of work already published that the intravenous administration of 50 mug of Salbutamol brought about a lowering of resistance in the placental blood vessels, and probably an increase in the blood flow at the maternal placental site, whereas, on the other hand when 500 mug of Salbutamol were injected intravenously the result was a deficiency in placental blood flow.


[Chromosome abnormalities in male sterility]. A study of the caryotype in 281 cases of male secretory sterility. Chromosomal anomalies were found in 81 cases, i.e. 78 cases, of azoospermia, 3 cases of oligospermia. Barr bodies were found in 66 cases of Klinefelter syndrome or one of its varieties. In the remaining 15 cases, there was no Barr chromatin: 7 of them bore gonosomal abnormalities with or without mosaicism. In 8 cases reciprocal translocations were found: in 2 of them a sexual chromosome was involved. The high rate of chromosomal anomalies should be stressed: every male secretory sterility, especially with azoospermia, warrants a chromosomal study. In the group of translocations, FSH and LH rates are not elevated in spite of the existence of azoospermia, whereas they are consistently increased in all the other chromosomal abnormalities: this so far undescribed phenomenon has no present explanation. The role played by translocations in male secretory sterility, although difficult to understand, is well documented, since translocations occur ten times more frequently in male sterility than in the general population.


Acuariid, capillariid and hymenolepidid parasites of the dasyurid marsupial Antechinus stuartii Macleay, 1841, from southeastern Australia. Stammerinema suffodiax sp. n. is described from the dasyurid marsupial Antechinus stuartii. It differs from S. soricis, the only other member of the genus, in lacking lateral body spines, as well as in the spicules and characters of the anterior extremity. As well as redescribing Synhimantus australiensis (Johnston and Mawson, 1952) now collected from the stomach of A. stuartii, a new capillariid, Capillaria rickardi from the stomach and two cestodes Hymenolepis aklei sp. n. and H. bradleyi sp. n. are described from the same host.


[Disorders of genital positioning. Attempted definition and classification]. We propose to classify in one chapter the different troubles of positioning which the uterus can have. In its site the uterus normaly has an extrinsic positions situation and position. The situation can be defined on the sagittal plane of the pelvic curve by the level in height of the organ in relationship to the bony pelvis. The position of the uterus is a global localisation of the uterus in the antero-posterior and frontal planes. The uterus also has an intrinsic status for which we had to have two terms of definition, the one being attitude and the other "bearing". The attitude is determined by the general morphology of the organ and the bearing is determined by the angle that the uterus makes in relationship to the axis of the pelvic cavity. In this way we can define exactly any position the uterus may take abnormally within precise groups.


Quantitative studies on Heterakis gallinarum infections in the common fowl, Gallus gallus L. The fertility, mortality, and migration patterns of Hetarakis gallinarum were studied in chickens with concomitant Parahistomonas wenrichi infections. H. gallinarum females were found to produce approximately 936 ova per day, when 50 days of age, and a total of 34,000 to 86,000 ova in a lifetime. There was no evidence of differential mortality between the sexes, nor of a preference for either the left or the right caecal organ of chickens. Both male and female worms are capable of migrating between caeca, and are expecially prone to do so when in the absence of individuals of the opposite sex.


[Pheochromocytoma and pregnancy].

[Pheochromocytoma and pregnancy]. The authors have taken the opportunity presented them by having a case of pheochromocytoma observed and diagnosed in the puerperium, of reviewing the various clinical and diagnostic features appropriate to this chromaffin tissue tumour. They also review the cardinal points in the differential diagnosis of any case of arterial hypertension discorvered in connection with pregnancy. A study of the literature shows that failure to recognise pheochromocytomata in pregnant women is accompanied by particularly hight maternal and fetal mortality. This complication of pregnancy, although rare, should be recognised by the obstetrician, who will be in a much better position early to combat and palliate the very grave threat to the mother and her baby.


[Pain in obstetrics].

[Pain in obstetrics]. In this article the authors start by going over the anatomical and neuro-physiological pathways of transmission of painful nerve impulses from the periphery to the centres. Pain is a "sensation" which is integrated in both a psychological and an emotional context. Psycho-physiological studies show that the cortex of the brain therefore plays a basic role in this integration and this allows us to consider the psycho-prophylaxis of delivery. Certain failures of psychoprophylaxis demonstrate the role of cultural factors and of factors that are specific to the subject of the pain of labour. The latter are dominated by anxiety, wich is at the same time due to the circumstances of the labour and of the individual patient. The latter stem from several sources: fear of death, separation, guilt and castration. There is one more psychological element that seems to be equally important and that is, morbid narcissism. The obstetrician should fight against the patient's anxieties which will thwart his work and disturb the onward progress of the labour. He will usually succeed but not always by the pathways that he thinks he is taking.


The effect of sublethal concentrations of the molluscicide niclosamide on the infectivity of Schistosoma mansoni cercariae. Experiments were conducted to assess the effect of sublethal concentrations of niclosamide on the infectivity of Schistosoma mansoni cercariae. Exposure of cercariae to 0.02 mg/l and 0.05 mg/l of niclosamide, respectively, for only two hours increased their mortality during penetration of mammalian host skin. The observed increase in mortality in the skin resulted in a consequent reduction of adult worm recovery from the liver and mesenteric veins of animals infected with the treated cercariae.


The morphology of Bothriocephalus scorpii (Müller) (Pseudophyllidea, Bothriocephalidae) from littoral fishes Britain. Observations have been made on the morphology of Bothriocephalus scorpii from littoral fishes in Britain, with particular reference to the reproductive system and the ultrastructure of the tegument. Two different kinds of microtriches, and ciliated sense organs, have been found. The shedding of proglottides by pseudoapolysis is described.


[Use of diazepam during labor].

[Use of diazepam during labor]. The effect of diazepam has been watched and studied by continuing monitoring of the heart rate of the mother, the fetus and the newborn, as well as by studying the acid-base balance of the mother and the fetus and the newborn after doses of 20 mg of diazepam had been given to 40 normal labouring women. A good calming effect was found in 87 p. % of women and the remainder had an acceptable result. Both in the mother and in the fetus tachycardia resulted from injection of diazepam as well as a lessening in the frequency of changes in the heart rate. These two alterations were statistically significant (p less than 0.01) and appeared very soon after the administration of the diazepam. Later, no other effects could be found in the mother, the fetus or the newborn.


Studies on the Malayan forest rat filaria, Breinlia booliati: periodicity and microfilaraemic patterns during the course of infection. Breinlia booliati exhibited nocturnal subperiodicity in its natural host, Rattus sabanus in contrast to experimentally infected laboratory-reared albine rats which showed irregular fluctuations of microfilariae throughout the 24 hour cycle. All the infected albino rats showed a prepatent period between 11-14 weeks postinoculation. Three patterns of microfilaraemia were discerned during the course of infection 38/49 rats displayed a single peak, 4/49 displayed 2 peaks about 12-15 weeks apart and 7/49 showed a sustained high plateau-like pattern of microfilaraemia. Cortisone had no effect on microfilarial levels when administered to rats near postpatency and some at postpatency.


[Threatened premature labor. Account of our recent experience]. We have been using two products, one an alpha-blocker (Vadilex) and the other a beta-stimulating drug (Ritodrine) to treat cases with threatened premature labour. Our experience is with 93 women who were treated in a continuous series. The overall result of our treatment show that 72 per cent of the cases were treated successfully and 28 failed. Analysing these preliminary figures in greater detail we have observed that the prognosis depends mainly on the state of the cervix at the time treatment is started in cases of threatened premature labour. The chances for success are much greater if the cervix is long and closed that if it is taken up or open. Since the trial was not carried out with a "double blind" control, we cannot really tell which of the two products we used was more efficacious.


Gazellostrongylus lerouxi Yeh, 1956 from antelopes in East Africa and its relation to the genus Paracooperia Travassos, 1935. A detailed redescription of Gazellostrongylus lerouxi and Paracooperia serrata was given from specimens collected from antelopes in Kenya. It was felt that there was no justification for the synonymy of the genus Gazellostrongylus with Paracooperia. Gazellostrongylus was placed in the tribe Ostertagiea.


Nasitrema attenuata (Trematoda) infection of Tursiops truncatus and its potentialities as an aetiological agent of chronic pulmonary lesions. Two cases of Nasitrema sp (Trematoda) infection of the bottlenosed dolphin, Tursiops truncatus, are reported and N. attenuata is briefly redescribed. The adults of the trematode occurring in the nasal sinuses and posterior nasal passage of the dolphins are considered as practically harmless for the host but thier eggs, aspirated deep into the bronchial tree, may initiate a foreign-body of inflammatory reaction in the lungs and continuous aspiration of such eggs may provoke a chronic pneumonia condition.


The growth of the metacestodes of Taenia crassiceps in white mice. The effect of the sex of the intermediate host on the growth and development of the metacestodes of Taenia crassiceps has been investigated. Three different strains of mice were used, the C R S, L A C A and C F L P strains and two strains of metacestode, the Toi and the E R S strains. The results show that matacestodes of T. crassiceps grow and multiply more rapidly in female than in male mice, although both sexes are equally susceptible to the infection. The origin of the parasite, i.e., from a rat or mouse host, affects the parasite growth.


Improvement in measuring migration areas in migration inhibition assays. An improved method for measuring migration areas in migration inhibition assays is described. In this method, drawings of the projected image are analysed with an image analysis system. This method is faster as well as more accurate than a standard method.


The use of 2-deoxy-D-glucose to assess changes in tumor target cell membranes in vitro. Following short-term suspension culture, cells from the Balb/C sarcoma Meth A were allowed to incorporate both [14C] leucine and 2-deoxy-D-glucose-1-[3H] (2DG). The 2DG is trapped as a small anionic marker of the cytosol. Deviation from the kinetics of spontaneous efflux of the markers is interpreted as reflecting perturbation of the target cell membrane. In the presence of guinea pig complement and a rabbit antiserum to Meth A, enhanced 2DG efflux was effected in a titer comparable to that detected with a 51Cr-release assay. With a number of alloantisera and syngeneic immune sera, 2DG efflux was enhanced while 51Cr-release was unaffected. Only in the presence of syngeneic immune sera from mice bearing a low tumor mass, syngeneic splenic leukocytes effect a retardation in the spontaneous 2DG efflux. Sera from animals with a large tumor mass were ineffective. Effux of proteins labeled with [14C] leucine was not altered. The phenomenon was not dependent on the presence of a heat-inactivatable syngeneic complement source. The method described provides a sensitive probe of target cell membrane permeability in the tumor model studied. The phenomenon detected is the capacity of serum, sampled relatively early in syngeneic oncogenesis, to direct syngeneic splenic leukocytes to interact with the target cell membrane differentially altering its permeability to the small cytosol marker.


The application of radiolabelled HL-A antibodies in the detection of (soluble) transplantation antigens: an alternative for the microcytotoxicity test. The use of radiolabelled anti HL-A antibody preparations is described. Absorption by platelets, followed by acid elution, fractionation of the eluate by column chromatography, radiolabelling and a second absorption--elution step with platelets yielded specific preparations, irrespective of original cytotoxic titres. The binding of these radiolabelled antibodies to lymphocytes could be inhibited by soluble antigens present in normal plasma or obtained from spleen cells by papain treatment. As a method for the detection of soluble transplantation antigens the use of radiolabelled antibodies appeared as sensitive as the microcytotoxicity test.


An indirect radioimmunoassay for mouse casein using 125I-labeled antigen. A new indirect radioimmunoassay was developed for detection of casein in mouse milk and in mammary tissue extract. Preincubation of rabbit gamma globulin to mouse milk casein (Ca2+-rennin precipitate) with unlabeled casein, milk, extracts of mammary tissues of late pregnancy and lactation, virtually blocked subsequent binding of 125I-labeled mouse milk casein to the antibody. Preincubation with mouse serum, bovine serum albumin, rennin, extracts of liver or immature mammary tissue had little effect on [125I]casein binding to the antibody. The inability of [125I]casein to bind to the antibody after preincubation with protein samples, which are likely to contain casein, is indicative of a specific antigen-antibody reaction. The assay is capable of detecting 0.2 mug casein, 1 mug milk proteins and 10 mug lactating mammary tissue extract. The application of the assay was also demonstrated using organ culture of the entire mammary gland. The glands treated with the lactogenic hormones, insulin + prolactin + cortisol, showed a saturation level of antibody-antigen reaction, indicating hormonal induction of casein; whereas, no reaction was observed with the non-treated gland.


Disturbance of hapten-antibody equilibria by ammonium sulphate solutions. A source of error in antibody affinity determinations. The affinity of anti-hapten antibody can be conveniently measured by precipitating immune complexes with ammonium sulphate. The method has, however, not proved very reproducible. Here is described one variable difficult to control in the assay: the ammonium sulphate was found to cause dissociation of ligands from hapten (NIP)--antibody complexes. The reason was the volume increase caused by addition of ammonium sulphate. The study suggested that in the calculation of the free hapten concentration the final volume during precipitation should be used. The precipitate should not be washed when hapten binding capacities are measured.


Identification of immunoprecipitates in line immunoelectrophoresis. A technique has been developed for the identification of immunoprecipitates in complex line-immunoelectrophoretic patterns. It is based on localized adsorption of individual antigens by monospecific antisera. The technique is suited for the following purposes as well: (1) testing the specificity and titer of uncharacterized antisera and (2) evaluation of binding properties of chromatographic media.


Radioimmunoassay for nanogram quantities of DNA. A direct competitive binding radioimmunoassay for DNA has been developed, using 125I-iododeoxyuridine-labelled DNA as the antigen and the serum from a patient with systemic lupus erythematosus. The assay is sensitive in the range of 25 to 1000 ng/ml of DNA. The sensitivity is determined by the affinity of the antibody: this SLE serum contains a component with an association constant of 9.6 X 10(-5) l/mol active at high dilution (1/10,000). Any biological material, such as serum, synovial fluid or tissue extracts can be tested directly. No interference has been found by DNAse in normal serum, or inhibition by mononucleotides or RNA. Native or denatured DNA from different sources (Escherichia coli, salmon sperm, calf thymus and human placenta) either purified or not, competes equally well for the antibody in this system.


Liposome spin immunoassay: a new sensitive method for detecting lipid substances in aqueous media. A new sensitive immunoassay procedure is described for quantitative detection of glycolipids and other lipids in aqueous media. As with other immunoassays specific antiserum is first reacted with the free lipid hapten. The amount of antibody activity remaining is measured by assaying the release, in the presence of complement, of spin label marker from liposomes containing the same lipid hapten. Using this method, 2.6 pmol of aqueous Forssman hapten was detected, and the sensitivity could be increased further.


Radioimmunoassay of plasma vasopressin in physiological and pathological states in man. A radioimmunoassay method for the measurement of arginine-vasopressin (AVP) in human plasma has been developed which requires 5 ml of plasma and has a lower limit of detection of 1-8 pg/ml plasma. Arginine-vasopressin was found to be stable in whole blood for up to 1 h at room temperature and for at least 4 h at 4 degrees C, while in plasma stored at -20 degrees C no loss was seen over 10 days. Dehydration and rehydration in normal subjects produced appropriate changes in AVP concentration but there was considerable variability in the levels attained by individual subjects and no obvious correlation with plasma osmolality. No consistent increase in plasma AVP concentration was seen on change of posture from the recumbent to the upright position. Vigorous exercise produced a marked rise in plasma AVP concentrations in most subjects which could not be attributed simply to an increase in plasma osmolality. In fusion studies with Pitressin in normal subjects showed a mean half-life of 6-4 min with an overall plasma clearance rate of 8-5 ml/min/kg body weight and a mean volume of distribution of 5-33 l. In patients with a biochemical picture suggestive of inappropriate antidiuretic hormone secretion, markedly raised plasma AVP concentrations were found only in patients with bronchial carcinoma.


Distribution of testosterone and 5alpha-dihydrotestosterone in rat epididymis and their concentrations in efferent duct fluid. 5alpha-Dihydrotestosterone concentrations were found to be appreciably higher in the proximal portion of the rat epididymis than in the distal portion. Following ligation of the efferent ducts of a testis, the 5alpha-dihydrotestosterone concentration diminished in the proximal half but not in the distal half of the epididymis. Epididymal fluid and spermatozoa were washed out from minces of epididymal tissue. The numbers of spermatozoa present in the wash fluid and in the homogenate of washed minces of epididymal tissue were used as measures for the amounts of epididymal fluid present in these fractions. Of the total amount of 5alpha-dihydrotestosterone (30-2 ng/g tissue) in the proximal portion, 20-5 ng were localized in the epididymal fluid, whereas of the total amount of 5alpha-dihydrotestosterone (8-6 ng/g tissue) in the distal portion, 2-8 ng were found in the epididymal fluid. The amounts of testosterone present in the fluids of both proximal and distal portions were 1-2 ng/g tissue. In contrast to the epididymal fluid, efferent duct fluid had a high concentration of testosterone (28-8 ng/ml) and a low concentration of 5alpha-dihydrotestosterone (1-9 ng/ml). These data suggest that the fluid surrounding the spermatozoa in the testis and the epididymis contains a high concentration of androgen and that as the fluid moves from the testis to the epididymis there is a clear change in the ratio of testosterone to 5alpha-dihydrotestosterone.


A comparison of variations in plasma luteinizing hormone concentrations in male and female domestic chickens (Gallus domesticus) from hatch to sexual maturity. Changes in plasma LH concentrations were followed in chickens of both sexes from hatch to sexual maturity using a radioimmunoassay. Mean levels of LH were lower in the females than in the males at all stages of development. These levels rose rapidly in both sexes during the first week after hatch to maxima of 6-5 +/- 1-2 (S.E.M.) ng/ml (n = 6) in the males and 4-6 +/- 0-6 ng/ml (n = 6) in the females. Thereafter levels of the hormone in the circulation stabilized in the males but fell over a period of 1 or 2 weeks in the females to 2-5-3 ng/ml. Plasma LH levels started to rise steeply in both sexes when they were between 16 and 19 weeks old at the same time as there was an increase in the rate of comb growth. Afterwards in six of the males studied in detail the mean plasma LH level rose significantly (P less than 0-01) over a period of 5-8 weeks from 8-1 +/- 1-2 to 13-2 +/- 1-9 ng/ml. In a parallel study on six females the rate of LH secretion increased for approximately 3 weeks and then decreased for about the same period forming a prepubertal LH peak. The first eggs were laid between 22 and 25 weeks of age when mean plasma LH levels had fallen to about 1-8 ng/ml. The mean plasma LH level in these hens when they were laying (1-8 +/- 0-3 ng/ml) was significantly lower (P less than 0-01) than when they were sexually immature (2-7 +/- 0-3 ng/ml). The duration of the period of rapid comb growth in each bird was closely related in the males to the time during which prepubertal LH peak. Differences in mean plasma LH concentrations in individual birds of either sex before the onset of puberty appeared to be related to subsequent reproductive performance.


Progesterone, prostaglandin F2alpha, and oestradiol in the utero-ovarian venous plasma of non-pregnant and early, unilaterally pregnant guinea-pigs. Levels of progesterone, prostaglandin F2alpha (PGF2alpha) and oestradiol in the utero-ovarian venous plasma of non-pregnant and early, unilaterally pregnant guinea-pigs were compared. Progesterone levels fell from day 12 to day 15 in the non-pregnant animals, while levels of PGF2alpha and oestradiol increased during this period. In contrast, in the pregnant guinea-pigs, progesterone levels remained high up to day 15 and PGF2alpha and oestradiol levels remained low in plasma from both the pregnant and non-pregnant sides. These findings support the hypothesis that the guinea-pig conceptus secretes an anti-luteolytic factor in early pregnancy, which reduces the amount of PGF2alpha produced by the uterus and released into the uterine veins and thereby allows luteal maintenance and continued progesterone secretion. Oestradiol, in the presence of progesterone, is regarded as the physiological stimulus for normal uterine PGF2alpha synthesis and release. Consequently, the inhibition of ovarian oestradiol secretion seen in early pregnancy is probably at least part of the mechanism by which the anti-luteolytic factor produced by the conceptus reduces PGF2alpha synthesis by and release from the uterus.


Effects of prostaglandin F2alpha and E2 on the production of progesterone by human granulosa cells in tissue culture. Human granulosa cells with differing steroidogenic potentials were cultured in vitro. The effects of prostaglandin F2alpha (PGF2alpha) and PGE2 on the progesterone output and viability of these cells were investigated. Prostaglandin F2alpha either alone or in combination with LH and FSH inhibited the production of progesterone over a wide range of concentrations (1-8000 ng/ml). However, the inhibitory effect of PGF2alpha was 200 times less effective when the cells were exposed to LH and FSH for 6 days before the addition of the prostaglandin. By contrast PGE2, at concentrations from 1 to 500 ng/ml, markedly stimulated the production of progesterone by granulosa cells, and this was not prevented by the addition of PGF2alpha. The degree of inhibition by PGF2alpha or stimulation by PGE2 was related to the biosynthetic capacity of the cells. These studies suggest that PGF2alpha may act directly on the adenylate cyclase system of human granulosa cells by blocking its activation by LH, and they demonstrate that functional regression of the luteal cell can be induced independently of the blood vascular system.


An unusual pattern of testosterone metabolism in tissue associated with a keratin-filled cutaneous cyst. Testosterone metabolism was studied in tissues associated with a keratin-filled cutaneous cyst. Instead of the 5alpha-reduced metabolites usually associated with skin steroid metabolism, considerable amounts of 5beta-reduced steroids were found. These included 5beta-androstane-3,17-dione, 17beta-hydroxy-5beta-androstan-3-one, and 5beta-androstane-3beta,17beta-diol. This change in metabolic pattern is discussed.


Presence of sex pheromone in preputial glands of male rats. Female rats consistently preferred the odour of male rat preputial gland compared with that of foot pads, submaxillary-sublingual glands, coagulating glands, liver, fat or muscle. Both saline homogenates and ether extracts were effective. Female rats did not respond to the odour of female preputial extract and they preferred the odour of normal male preputial extract to that from castrated rats. The pheromone was not associated with the fatty acids of the preputial extract. The fractionation of the volatile components of preputial extracts by gas chromatography revealed that most of the biological  activity resided in a specific fraction.


Rapid interaction between luteinizing hormone and isolated rabbit follicles. The rapidity of binding of LH to rabbit ovarian follicles was examined using androgen production as the endpoint. Incubations were carried out for about 8 h with medium being changed every 15 min except during the periods when follicles were exposed to LH and/or anti-LH and subsequent washing. Exposure of follicles to LH for 1 s followed by washing in (a) LH-free medium, (b) pure anti-LH or (c) diluted anti-LH, failed to inhibit androgen production which persisted for more than 2 h. Simultaneous addition of anti-LH and LH to the medium before incubation with follicles completely inhibited the response though it could be elicited from the same follicles when LH was added later. These data suggest that the binding of LH to follicular receptors is a rapid process and that either the LH-receptor complex is very stable or that LH initiates a series of reactions which are independent of the presence of hormone.


Specific release of bovine neurophysin II during arterial or venous haemorrhage in the cow. An almost specific release of neurophysin II with only minor release of neurophysin I was found during venous or arterial haemorrhage in five cows. Our data confirm the results of experiments in vitro in which vasopressin has been found to be associated with neurophysin II within neurosecretory granules.


Enzyme activity in kidney, adrenal and gonadal tissue of rats treated neonatally with androgen or oestrogen. A single injection of 300 mug oestradiol benzoate (OEB) or 1.25 mg testosterone propionate (TP) on day 1 of life led to significant changes in the activity of enzymes involved in steroid hormone metabolism in kidney, adrenal and gonadal tissues of adult rats. In the kidney, the enzyme activities of male rats reacted to OEB, but not TP, by the development of normal female levels. With one exception the enzyme activities of the kidney of female rats did not respond to either steroid. In the adrenal of both sexes 5alpha-reductase reacted to OEB, but not TP treatment, by a fourfold increase in activity. In the ovary all the enzymes investigated responded both to OEB and TP treatment by a fall in activity; 20alpha-hydroxysteroid dehydrogenase activity fell to undetectable levels. In the testis, OEB and TP treatment led to contrasting effects. With the exception of 5alpha-reductase all the enzymes tested in this organ responded to OEB by a rise in activity. Where TP had any effect, it produced a slight decrease in activity.


Sexual behaviour in castrated rabbits treated with testosterone, oestradiol, dihydrotestosterone or oestradiol in combination with dihydrotestosterone. Sexual behaviour and the function of the accessory sexual glands were studied in castrated rabbits injected with testosterone benzoate (TB), oestradiol benzoate (OEB), dihydrotestosterone benzoate (DHTB) or OEB in combination with DHTB. Testosterone benzoate (1 mg daily for 90 days) stimulated the sexual behaviour more than any of the other steroids. The combination of OEB (0-33 mg) and DHTB (1 mg) was no more effective than either of these steroids given alone. The function of the accessory sexual glands was stimulated to a level comparable to that of intact animals given TB. Dihydrotestosterone benzoate was, however, not very effective in this respect. Oestradiol benzoate alone or in combination with DHTB caused hypertrophy and very low secretory activity of the seminal vesicles. These results suggest that testosterone itself is active both in the brain and in the accessory sexual glands in rabbits. This is in contrast to the rat, in which aromatization to oestradiol in the brain and reduction to DHT in the periphery seems to be important.


Protein binding of plasma cortisol in the foetal lamb near term. Binding of cortisol to plasma proteins was studied in the foetal lamb by equilibrium dialysis at 37 degrees C. At 122 days of pregnancy the mean level of transcortin expressed as cortisol-binding capacity was 28 +/- 6 (S.D.) ng cortisol/ml plasma. During the last 14 days of pregnancy there was a progressive increase in transcortin-binding capacity to 85 +/- 14 ng cortisol/ml plasma. A sharp increase in the concentration of both protein-bound and unbound cortisol was observed over the same period. A rise in the concentration of total cortisol from around 3 to 42 ng/ml was associated with an increase in unbound cortisol from 0-2 to a maximum of 2-1 ng/ml. The concentration of albumin-bound cortisol was approximately equal to that of unbound cortisol. The mean value for the transcortin-cortisol affinity constant was 1-15 x 10(8) l/mol. It is concluded that an increase in transcortin-binding capacity is partly responsible for the prepartum increase of corticosteroid levels observed in normal foetal lambs.


Effects of injections of monoamine oxidase inhibitor or saline into the uterus in late pregnancy on uterine catecholamine levels related to abnormal parturition in rats. Injection of a monoamine oxidase (MAO) inhibitor (nialamide) into the uterus of an anaesthetized and laparotomized rat on day 20 of pregnancy severely disturbed parturition. Injection of the solvent (0-9% isotonic NaCl solution) at the same stage of gestation produced the same but less frequent disturbances. When the rats were injected on days 19 or 21, impairment was less marked than on day 20. Therefore, day 20 seems to be a critical period for the onset of parturition. Injection of Ringer solution into the uterus on day 20 had effects analogous to those of saline injection at the same stage. Anaesthesia induced with ether, laparotomy of the pregnant rat on day 20, and handling of the uterine horns without injection of either Ringer or NaCl also disturbed parturition in 70% of the rats treated. Nevertheless, disorders were not as severe as those after injection. Laparotomy alone on day 20 did not disturb parturition. The effects on parturition of a saline injection into the uterus on day 20 were greatly decreased when the injection was performed on pregnant rats adrenalectomized on day 14, or on pregnant rats pretreated on days 18 and 19 with an agent blocking the adrenergic beta receptors (propranolol); 70-80% of the treated rats had normal deliveries. In control rats, uterine catecholamine levels were markedly modified between days 21 and 22 of gestation. These changes did not occur in rats injected with MAO inhibitor or saline.


Effect of L-dopa on milk ejection and prolactin release in lactating rats. The effect of L-DOPA on milk ejection and on prolactin release during 30 min of suckling was studied in lactating rats. Various doses of L-DOPA (1-25, 2-5, 5 and 10 mg/100 g body wt) were injected i.p. 30 min before the suckling period. Control rats were injected with 0-9% NaCl solution only. An inhibition of milk ejection proportional to the dose of drug administered was obtained. The dose of 10 mg completely blocked milk ejection but 1-25 mg had no effect. A normal milk-ejection response was obtained with a small dose of oxytocin injected immediately before nursing into mothers treated with 10 mg L-DOPA, indicating that the blocking effect was not due to a lack of mammary gland response. In control mothers, serum prolactin levels increased from 67-2 +/- 25-9 (S.E.M.) to 950-3 +/- 118-7 ng/ml after a 30 min suckling period. L-DOPA (5 and 10 mg) prevented the release of prolactin induced by suckling, but 1-25 and 2-5 mg L-DOPA had no effect. The results indicate that oxytocin and prolactin release induced by suckling in lactating rats is inhibited by an increase of catecholamines at the hypothalamic-hypophysial axis.


Plasma luteinizing hormone and testosterone in the adult male pig: 24 hour fluctuations and the effect of copulation. Secretory patterns of LH and testosterone were characterized in the intact male miniature pig. All blood samples were taken from indwelling catheters. Hourly sampling was carried out over 24 h and during a morning period blood was collected for 2 h at 10 min intervals. No significant difference was detected in the plasma LH concentration on the basis of hourly sampling. Plasma testosterone was significantly (P less than 0-05) lower during the evening and night when compared with morning values. The second experiment was concerned with the pattern of plasma LH and testosterone concentrations before and after copulation. Blood sampling was performed at 10 min intervals. Plasma LH was significantly (P less than 0-001) raised for 30 min after copulation when compared with any 30 min period (0-120 min) before copulation. Plasma testosterone was not significantly altered for any 30 min period of the experiment (0-270 min). The data are interpreted as a possible mechanism for endocrine control of testicular function.


Electrical stimulation of the hypothalamus and luteinizing hormone secretion in Japanese quail. Experiments were undertaken to localize those hypothalamic areas in the male quail (Coturnix coturnix japonica) where electrical stimulation would increase LH secretion. The posterior basal hypothalamus was stimulated with rectangular pulses (height 500 muA) through a bipolar electrode for 6 min, blood samples being taken for LH assay 20 min before, and 2, 10, 20 and 30 min after stimulation. The highest plasma concentration was observed in the 2 min sample. Over the next 30 min the LH level decreased to the resting concentration. The relative increase in LH level was greatest in sexually immature quail and least in photostimulated castrated birds, although the highest absolute levels were seen in the castrated quail. There were no statistical differences between the magnitude of the LH increases in sexually immature, mature and castrated quail. Various hypothalamic regions were then stimulated with a smaller current (200 muA) applied for only 2 min. A highly significant rise in LH followed stimulation of either the tuberal hypothalamus (postero-dorsal part of the infundibular nuclear complex, PD-INC), or the preoptic region (POR) while stimulation 0-5-1-5 mm away from these regions did not change LH secretion. Stimulation of the anterior basal hypothalamus, or of the suprachiasmatic area, caused a significant rise in LH concentration although this was less than that seen after stimulation of the POR. Stimulation in the POR or the PD-INC was ineffective if the tuberal hypothalamus had been deafferentated surgically some days previously. The data complement the studies in which destruction of the PD-INC or the POR by electrolytic lesions has been shown to block photoperiodically induced testicular growth and LH secretion.


Suppression of the development of female hamster behaviour by implants of testosterone and non-aromatizable androgens administered neonatally. Testosterone in its free form, and dihydrotestosterone (DHT) and androsterone, both androgens which are not aromatizable to oestrogen, injected in oil during the neonatal period have been reported not to modify the development of female sexual behaviour. This failure might be due to the short period of activity of these substances when injected in liquid vehicles. In the current study, a Silastic pellet containing 9% of its weight of testosterone, androsterone, or DHT was implanted subcutaneously in 42 female and 38 neonatally castrated male hamsters on day 2 of life and removed on day 10. Pellets of pure Silastic were implanted in 36 control animals. Males were gonadectomized on day 5 and females on day 45. Female sexual behaviour induced by oestradiol benzoate and progesterone was measured in a series of 10-min mating tests with vigorous males, starting at 55 days of age. The duration of lordosis was consistently reduced below control levels in females implanted with testosterone, DHT, and androsterone, and in males, with testosterone and DHT. Thus the free form of testosterone, and some non-aromatizable androgens, when present for a sufficiently long period after birth, can permanently suppress development of female reproductive behaviour.


The pigeon's perception of saturation.

The pigeon's perception of saturation. Three experiments used similar methods to investigate the pigeon's perception of saturation of monochromatic lights. This trial-wise procedure consisted of brief presentations of positive and negative stimuli in random sequence. Pecks to the positive stimuli were occasionally reinforced on a low fixed-ratio schedule. The first study determined absolute thresholds for "white" and monochromatic lights by establishing a discrimination between lights of various radiances and a dark key. Experiment II investigated generalization from a white light to various monochromatic lights under conditions that minimized the use of luminance as a cue. The third experiment examined discrimination of various monochromatic lights along a colorimetric purity continuum; responses to white light were reinforced, while responses to lights that combined white and monochromatic lights in various proportions were not. The results indicated that lights of different wavelength differ in saturation, but that all are discriminable from white. Wavelengths between 550 and 600 nm are least saturated for the pigeon, and saturation increases markedly as wavelength decreases below this region of the spectrum.


Transfer of hue matching in pigeons.

Transfer of hue matching in pigeons. Pigeons were trained on a modified three-key matching-to-sample procedure, in which only one comparison key (rather than two) was lighted after an observing response to the center-key standard. Pecks on keys of matching comparison hues were reinforced. When non-matching hues appeared as the initially lighted comparisons, the nonmatching hue terminated and the matching hue appeared on the other side key only if the pigeon did not peck the nonmatching comparison for 4.8 sec. Pecks to the nonmatching hue reset the 4.8-sec delay interval. Three hues were used during acquisition. During transfer tests, two novel hues were substituted individually or together for one or two of the training hues. Latencies to the novel side-key hue were shortest when a novel matching hue appeared as the standard on the center key, and were essentially identical to baseline matching latencies. In contrast, when a novel hue appeared as either a standard or comparison in a nonmatching combination, latencies increased with increasing separation between the noevel hue and the nonmatching hue. These transfer data demonstrate the concept of hue matching.


Effects of d-amphetamine, chlorpromazine, and chlordiazepoxide on intercurrent behavior during spaced-responding schedules. Effects of d-amphetamine, chlorpromazine, and chlordiazepoxide on lever pressing under direct control of spaced-responding schedules were compared with effects on intercurrent drinking and wheel running in the rat. Drug effects on lever pressing were systematically related to dose and were consistent for all animals; drug effects on intercurrent behavior were generally different for each animal. In the case of lever presses, increasing doses of d-amphetamine first increased and then decreased response rate, increasing doses of chlorpromazine produced graded decreases in response rate, and doses of chlordiazepoxide up to 40 mg/kg produced no effect on response rate. These data are discussed in context with the concept of schedule control, and it is suggested that the behavioral pharmacology of intercurrent behavior be explored as a useful procedure in the experimental analysis of intercurrent behavior.


The effect of temperature on the outward currents in the soma of molluscan neurones in voltage-clamp conditions. The delayed outward current in snail neurones was separated into two components with different temperature sensitivity: (i) a persistent component and (ii) a transient (inactivating) component. The effect of cooling on the value of the transient current is strongly dependent upon the value of the conditioning potential. It was supposed that cooling causes a decrease in the negative surface potential in the vicinity of the potassium pathways and removes their inactivation. Simultaneously cooling depresses the potassium conductance. The effect on surface potential is more distinct with conditioning potentials at which a significant fraction of the transient outward current is inactivated. The effect of cooling on the transient component of the fast outward current was similar to that on the transient component of the delayed outward current.


Peripheral control of acoustic signals in the auditory system of echolocating bats. Many species of echolocating bats emit intense orientation sounds. If such intense sounds directly stimulated their ears, detection of faint echoes would be impaired. Therefore, possible mechanisms for the attenuation of self-stimulation were studied with Myotis lucifugus. The acoustic middle-ear-muscle reflex could perfectly and transiently regulate the amplitude of an incoming signal only at its beginning. However, its shortest latency in terms of electromyograms and of the attenuation of the cochlear microphonic was 3-4 and 4-8 msec, respectively, so that these muscles failed to attenuate orientation signals by the reflex. The muscles, however, received a message from the vocalization system when the bat vocalized, and contracted synchronously with vocalization. The duration of the contraction-relaxation was so short that the self-stimulation was attenuated, but the echoes were not. The tetanus-fusion frequency of tha stapedium muscle ranged between 260 and 320/sec. Unlike the efferent fibres in the lateral-line and vestibular systems, the olivo-cochlear bundle showed no sign of attenuation of self-stimulation.


Transport of amino acids by isolated gills of the mussel Mytilus californianus Conrad. The unidirectional influx of cycloleucine into in vitro preparations of gill tissue of the mussel, Mytilus californianus, was determined. Influx was found to be linear for at least an hour, and the kinetics of cycloleucine influx conformed to Michaelis-Menten type kinetics. The transport mechanism(s) for cycloleucine is relatively specific for the L-enantiomorph of neutral amino acids, and is capable of accumulating cycloleucine to intracellular concentrations much higher than those of the surrounding medium. Evedence is presented that the transport of amino acids by gill tissue plays a significant role in whole animal nutrition.


Factors affecting the heart activity and blood pressure of the swan mussel Anodonta cygnea. 1. The rate of heart beat increased with temperature and was three times as high in the active as in the inactive animal. 2. The rate of shell valve movement rose and the rate of heart beat fell when the foot was extended. 3. The rates of heart beat and shell valve movement decreased when the water was saturated with carbon dioxide. This heart response remained when the visceral ganglion was destroyed. 4. Ventricular contraction occurred simultaneously over the shole chamber. The passage of blood into the posterior aorta could be restricted by the protuberances on its wall. 5. Pericardial cavity pressure rose by about 5 cm H2O at shell valve adduction and 0-25--0-6 cm H2O at ventricular diastole. 6. Pulse pressure changes of 0-25--0-6 cm H2O occurred in the auricle and 1--3 cm H2O in the ventricle and anterior aorta.


Role of active potassium transport by integumentary epithelium in secretion of larval-pupal moulting fluid during silkmoth development. 1. The exuvial side of the pharate pupal integument is usually positive to the haemolymph-side, both in vivo and in vitro, during the period when the moulting fluid is being secreted. 2. The ratio of potassium flux toward the exuvial space is higher than that toward the haemolymph, under both open-circuit conditions and short-circuit conditions, demonstrating by the Flux Ratio test that potassium is actively transported across the isolated integument during this secretion period. 3. Just prior to ecdysis, while moulting fluid is being reabsorbed, the potassium flux ratios become unity, suggesting that active potassium transport has ceased, but the short-circuit current that remains suggests that some other ion is actively transported at this time. 4. We argue that the potassium salt solution, formed in the exuvial space (as water presumably follows the actively transported potassium), has three functions (1) to accomplish the gel--sol transformation, (2) to activate the gel enzymes and (3) to buffer the enzyme solution at a pH favourable to the activity of the gel enzymes.


Modes of stimulation of the teleost ear. Microphonic potentials were recorded from the inner ears of a catfish (Ictalurus punctatus) and an African mouthbreeder (Tilapia macrocephala) in response to underwater sound stimulation and direct vibration of the head. The shape of the vibratory isopotential functions of frequency was similar in both species up to 600 HZ. Above 600 HZ, the sensitivity of Ictalurus continued to increase to 4000 HZ while the sensitivity of Tilapia declined. Deflation of the swim bladder did not affect the response to vibration in either species, the response of Tilapia to the underwater sound stimulus being minimal and unaffected by removal of the swim bladder. Ictalurus was pressure-sensitive to above 4000 HZ, ther being a significant dedline in the response with deflation of the swimbladder.


An in vitro bioassay for a mulluscan gonadotropin. 1. Protein synthesis occurs at a high rate in the ovaries of maturing Octopus vulgaris and can be measured from the incorporation of [14C]leucine in vivo and in isolated groups of eggs in vitro. 2. Removal of the optic glands in vivo 1--3 days prior to testing markedly reduces amino acid incorporation in vivo or in vitro. After 5 days in vivo incorporation stops. 3. The rate of incorporation in vitro is increased by the addition of optic gland extract. 4. Analysis of the kinetics of leucine uptake and incorporation in vitro indicates that the hormone has an effect on the inward transport of leucine which is independent of its action on protein synthesis. 5. Electron-microscope studies of the follicle cells and ova show that the former are the site of protein synthesis. 6. Changes in either uptake or incorporation into protein by the follicle cells can be used as a qualitative biolobical assay for the optic gland hormone. Uptake is very easy to measure but incorporation is the more sensitive parameter. Either is potentially suitable as a quantitative assay for this and perhaps also for other molluscan gonadotropins.


Activity and habituation in the brain of the polyclad flatworm Freemania litoricola. 1. A variety of spontaneously active units was measured in the brain of the polyclad flatworm Freemania litoricola. Following application of MgCl2 there was both a decrease in number of active units and a decrease in frequency of firing of those cells which persisted in their activity. 2. Receptors which respond to vibration stimuli evoke potentials in the posterior part of the brain. Repetitive stimulation leads to habituation, the extent of which is dependent on both the number of times stimulated and the strength of the stimulus. Weaker stimuli habituate more rapidly than strong stimuli. Habituated responses can be dishabituated by tactile stimuli and also by stronger intensity stimuli of the same modality. The vibration-evoked potentials appear to occur in at least second-order cells, since vibration responses are abolished by the application of MgCl2. 3. Tactile responses can also be elicited from the posterior portion of the brain when the stimulus is applied to the periphery of the animal. These responses are insensitive to MgCl2. 4. Both vibration and tactile evoked responses are able to evoke further barrages of spike activity. 5. The presence of a dual sensitizing and inhibitory system during habituation is discussed.


Electrophysiology of the peripheral nerve net in the polyclad flatworm Freemania litoricola. 1. A diffuse-conducting system close to the dorsal epithelium of the polyclad flatworm Freemania litoricola is described. Tactile stimuli elicit small action potentials which can be conducted around lesions through the body wall. The potentials can occur in bursts or barrages. 2. This conducting system appears to be insensitive to Mg2+ ions. 3. Conduction velocities (0-26--71 m/sec) vary over the animal. Conduction spread in the anterior half of the animal appears to be greater than that in the posterior portion. 4. Response decrement to repeated stimulation can be recorded in the peripheral system but it is not clear if this is due to habituation or fatigue. 5. Conduction from the peripheral net to the brain occurs. Some central units appear to pick up information only, or mainly, through the anterior nerves, while other units can respond to information conducted through the network to nerves of the contralateral side. 6. Different possibilities to account for this system are discussed, and it is suggested that the animals either possess a unique Mg2+ insensitive synaptic nerve-net or else the network is electrically coupled.


The cardioregulatory system of crayfish: neuroanatomy and physiology. 1. The anatomical arrangement of the cardioregulatory nerves and their physiological activity during cardiac modulation were analysed in Procambarus clarkii. 2. The bilaterally arranged pairs of cardioinhibitors and cardioaccelerator axons, in nerves SN II and SN III respectively, were physiologically identified by correlating spikes in SN II and SN III with the same spikes in the dorsal nerve, which innervates the heart. 3. The cardioinhibitor neurone fired tonically in varied sporadic bursts. During periods of cardiac inhibition, however, this neurone discharged in a long chain of spikes at a characteristic frequency of 40-50Hz. 4. The cardioaccelerator neurone fired tonically at 2-3 Hz but on occasion its activity reached 12 Hz. 5. Three inhibitory cardiac reflexes were analysed. The sensory modalities for the reflexes included (a) stretch of the dorsal pericardial wall, (b) chemical stimulation of coxal hair sensilla with glucose and (c) tactile stimulation of hair sensilla in and below the gill chamber, on the antennae, the antennules and on the anterior cephalothorax. 6. The discharge of both cardioinhibitor neurones showed a weak temporal correlation suggesting a common presynaptic drive, while the pair of cardioaccelerators appeared to have a reciprocal relationship with the cardioinhibitors.


The cardioregulatory system of crayfish: The role of circumoesophageal interneurones. 1. Interneurones located in the circumoesophageal commissures were found to control the activity of the cardioinhibitor and cardioaccelerator regularity neurones. 2. These interneurones (cardiac command fibres) fell into three classes: (1) strong inhibitors, which caused cardiac arrest, (2) weak inhibitors, which caused bradycardia, and (3) accelerators, which caused tachycardia. 3. When the positions of interneurones were plotted collectively, they formed distinct clusters, suggesting that each cluster was represented by one command unit in an individual preparation. Twenty strong inhibitor units and 16 accelerator units were found. Weak inhibitors did not form clusters. 4. Stimulus threshold characteristics were as low as 3.0 V and 3 Hz for the strong inhibitor units (mean range 4.7-7.1 V and 14-30 Hz). Higher values were found for weak inhibitors and accelerators. 5. The strong inhibitor command drives always showed a positive bias toward the contralateral cardioinhibitor neurone, relative to the ipsilateral cardioinhibitor. 6. Plots of command neurone stimulating frequency versus evoked cardio-inhibitor activity displayed steep positive slopes for strong inhibitor command units and shallow positive slopes for weak inhibitor units. 7. Reciprocity between the cardioinhibitor and cardioaccelerator neurones occurred during both inhibitory and acceleratory command drives. This is not likely to be a property inherent in the command units themselves because reciprocity was earlier observed during chemical and tactile reflex inhibition of the heart. (Field & Larimer, 1974a).


Nervous control of respiration: oxygen-sensitive elements in the prosoma of Limulus polyphemus. 1. Responses of oxygen-sensitive units in the prosomal haemal nerve of Limulus polyphemus were examined while varying the oxygen content of sea water bathing the intercoxal cuticle. 2. When exposed to high oxygen levels these units units maintained a continuous background discharge of spikes. Unit activity was inhibited when oxygen content decreased. Upon reintroduction of oxygen tonic spike discharge resumed. 3. Mechanosensitive units with receptive fields on the prosomal shield or intercoxal cuticle were also present in the haemal nerve. Neither the mechanosensitivity nor the background discharge of these units was affected by changes in oxygen content. 4. It is proposed that the oxygen-sensitive respiratory reflexes of Limulus are an adaption to existence in the tertidal zone. Published observations of the respiratory stress responses of many intertidal animals support this hypothesis.


Behavioural and electrophysiological studies of taste discrimination by the maxillary palps of larvae of Locusta migratoria (L.). 1. Behavioural studies show that larvae of Locusta migratoria (L.) can discriminate between certain simple chemicals and between chemicals obtained from plant sources. 2. Electrophysiological tests show that within each sensillum the same neurones respond to different chemicals. 3. The frequency of occurrence of sensilla with enhanced specificity to certain chemicals is investigated. 4. Statistical tests confirm that, despite the variability of response, across-fibre analysis could allow discrimination between chemicals provided an adequate number of senilla are used. 5. The conclusions are discussed in relation to current theories of chemoreceptor functioning.


Optic gland implants and their effects on the gonads of Octopus. 1. Optic glands transplanted from one Octopus vulgaris into another cause enlargement of the gonads and ducts of the recipient. 2. Enlargement occurs whether or not the gland was secreting when implanted and regardless of the sex of the donor or recipient. 3. Glands derived from Eledone moschata or Octopus macropus implanted into O. vulgaris are as effective as glands derived from O. vulgaris. 4. Implants derived from Sepia officinalis or Loligo vulgaris appear to be ineffective.