Scintigraphic assessment of vascularity and blood-tissue barrier of human brain tumours. Assessment of vascularity and blood-tissue barrier was performed by sequential scintigraphy in 43 patients with brain tumours. The blood-tumour barrier was evaluated by use of 99mTc-pertechnetate, and vascularity using 99mTc-labelled red blood cells. Three groups of tumours were found: tumours with low vascularity and permeable barrier, tumours with high vascularity and permeable barrier, and tumours with low vascularity and relatively impermeable barrier. The first group indicates that when vessels are permeable, there may be a rapid penetration of large amounts of pertechnetate into the tumour even when vascularity is not increased. In the other two groups penetration of pertechnetate into the tumour is affected by vascularity, as it determines the total area where passage of the radiopharmaceutical takes place. It is suggested that the permeability of the blood-tumour barrier and the amount of vascularity may have an effect on the success of chemotherapy in brain tumours.


Sinusoidal oscillation of the ankle as a means of evaluating the spastic patient. Sinusoidally modulated torque was applied to rotate the ankle joint of normal subjects and clinically spastic patients. Measurements were made of the effective joint compliance and of the evoked EMG activity. These procedures provide a well-quantified and reliable measure of muscle tone and hyperreflexia which is well correlated with the patient's clinical status.


Mechanism of monosynaptic reflex reinforcement during Jendrassik manoeuvre in man. The facilitating effect of the Jendrassik manoeuvre on monosynaptic reflexes in man has been studied by comparing H and T reflexes of the soleus and by blocking the effects of spindle activation by ischaemia of the leg. The Jendrassik manoeuvre equally enhances H and T reflexes provided that the test reflexes are small. The H reflex remains facilitated when the spindle activation cannot affect the soleus alpha-motoneurones--that is, when the Ia afferent nerve fibres from the soleus are blocked by ischaemia. It is concluded that the facilitating effect of the Jendrassik manoeuvre on the alpha-motoneurones is not predominantly routed via the gamma-loop.


F responses studied with single fibre EMG in normal subjects and spastic patients. Recurrent responses of single human motor neurones after antidromic activation, the so-called F response, have been studied in the abductor digiti minimi muscle in normal subjects and in spastic patients with the single fibre EMG technique. The F response occurs rarely and in groups. Half of all motor neurones did not produce any F response within an arbitrarily chosen time interval of 200 successive stimuli at 1 and 2 Hz. Spastic patients had as many silent neurones as the normal subjects but the responding neurones more frequently produced F responses (P less than 0.001). When a neurone has few responses at rest the frequency increases under activation of the contralateral or ipsilateral muscle (P less than 0.05). When there are many responses in the relaxed state there is a decrease (P less than 0.001) under activation. The behaviour of an individual neurone is consistent in different activation periods.


Refractory period studies in a human neuromuscular preparation. A subtraction technique has been employed to study refractoriness in the median nerve and thenar muscles in man. The absolutely refractory periods of the distal motor nerve branches and of the muscle fibres were found to be similar, the majority being within the 2.5--3.0 ms range; the values were distributed in a unimodal manner. The relatively refractory periods of the motor nerve terminals were about 5 ms. In some experiments it was possible to record non-propagated potentials at the endplate zones of the muscle fibres.


Effects of age, temperature, and disease on the refractoriness of human nerve and muscle. Absolutely and relatively refractory periods have been investigated in terminal median nerve axons and in thenar muscle fibres in a variety of pathophysiological conditions. Marked increased in refractoriness were produced by cooling to 20 degrees C. Nerve fibre refractoriness was more pronounced in older subjects and diminished in myotonic dystrophy. In contrast, no significant changes in muscle fibre excitability could be demonstrated in either condition or in patients with renal failure or motor neuropathies.


Aspects of interhemispheric subdural haematoma, including the falx syndrome. Two cases of interhemispheric subdural haematoma are reported. The first patient presented with a falx syndrome of contralateral hemiparesis, most marked in the lower extremity. The second patient had vertex headaches and was diagnosed with computerised tomography. Serial studies suggested that the interhemispheric haematoma may have migrated to a lateral position over the cerebral convexity.


Antenatal diagnosis of Krabbe's leucodystrophy: enzymatic and morphological confirmation in an affected fetus. Galactosylceramide beta-galactosidase activity was assayed in cultured amniotic fluid cells from two pregnancies at risk for Krabbe's leucodystrophy. The elective termination of one pregnancy was carried out after demonstration of a severe deficiency of galactosylceramide beta-galactosidase activity. The diagnosis of Krabbe's leucodystrophy in the affected fetus was confirmed enzymatically by a deficiency of galactosylceramide beta-galactosidase inbrain, liver, kidney, and cultured skin fibroblasts, and histologically by the ultrastructural demonstration of the typical intracellular inclusions in cells of the spinal cord.


Large suprasellar aneurysms imitating pituitary tumour. Two patients had large (20--30 mm) suprasellar aneurysms with clinical findings resembling an expanding pituitary tumour. One aneurysm was giant (30 mm). Its delayed diagnosis for six years led to severe chiasmal compression. The other aneurysm was compressing the optic nerve. Visual improvement resulted after surgery.


Calcified congenital arachnoid cyst with heterotopic neuroglia in wall. Two unique findings, advanced calcification and ectopic neuroglia, were encountered in the wall of a giant congenital arachnoid cyst occurring in a 40 year old woman. The cyst almost totally filled the supratentorial subdural space of the left hemicranial cavity, was not connected with the subarachnoid space, and thus developed intra-arachnoidally. Its congenitally derived nature was supported by the unique finding of heterotopic neuroglia in the wall. Congenital arachnoid cyst must be distinguished from various cystic lesions within the central nervous system, including the neuroepithelial cyst which can arise throughout the neuraxis. It is suggested that pathogenesis of the congenital arachnoid cyst is related to aberrant flow of the ventricular cerebrospinal fluid into the developing leptomeninx in the process of differentiation of the subarachnoid space. The tract or pouch may occur within the arachnoid mater, and the cyst is formed intra-arachnoidally when the former filled with fluid is closed off from the subarachnoid space.


Temperature-sensitive conduction failure at axon branch points. 1. The propagation of action potentials through the branching regions of squid axons was examined experimentally and with computer simulations over a temperature range of 5-25 degrees C. 2. Above a critical ratio of postbranch to prebranch diameters, propagation of an action potential failed. The value of this critical ratio is very sensitive to temperature and is smaller at high temperatures. The experimentally measured Q10 of the critical ratio is 0.37 +/- 0.04. 3. Evaluation of a number of parameters of action-potential propagation showed that this effect is closely related to the change in the width of the action potential with temperature (Q10 = 0.29 +/- 0.01).


Primary afferent depolarization and inhibory interactions in spinal cord of the stingray, Dasyatis sabina. 1. Excitability changes in primary afferents and inhibitory interactions in evoked spinal cord activity were investigated in unanesthetized stingrays (Dasyatis subina) with high cervical spinal transections. 2. Primary afferent excitability increases following a conditioning stimulus to an adjacent segmental nerve were demonstrated with the Wall (31) technique. 3. Stimulation of A-alpha,beta and A-delta afferent fibers produced excitability increases in both A-alpha,beta and delta-fibers of the adjacent segment. 4. The excitability increase had a latency of about 10 ms, it peaked around 25 ms, and the change lasted more than 100 ms. 5. The central afferent volley in A-alpha,beta fibers and the N1- and late negative waves due to postsynaptic activity of dorsal horn interneurons were reduced by conditioning volleys in adjacent afferent nerves. The time course of the inhibition paralleled that of the excitability increases in afferent terminal arborizations, suggesting that the depression of postsynaptic activity is, at least in part, due to presynaptic inhibition. 6. Reduction of evoked discharges and excitatory postsynaptic potentials was observed in recordings from interneurons with a time course similar to that of the primary afferent depolarization (PAD). 7. Conditioning volleys in afferents of adjacent peripheral nerves produced facilitation or inhibition of segmental reflexes.


Response of cutaneous hair and field mechanoreceptors in cat to paired mechanical stimuli. 1. While studying hair- and field-receptor responses to paired mechanical pulses, I observed that in approximately half the receptors the recovery of excitability following generation of an impulse took the form of a damped oscillation. 2. Oscillations could be modified or eliminated by changing the conditions of stimulation: 1) there were no oscillations with an electrical conditioning stimulus; 2) oscillations occurred only when both mechanical pulses were delivered to the surface of the skin, not when the conditioning pulse was given to a hair shaft, and only when both pulses affected the same part of the receptive field; 3) oscillation was reduced by increasing the duration of the pulses; 4) oscillation was enhanced by increasing the distance between the stimulator and the nerve terminal; and 5) maxima on the recovery curve became minima and vice versa when the direction of the conditioning pulse was changed. 3. It was concluded that recovery curve oscillations were due not to the type of hair or field receptor being stimulated, but to the conditions of stimulation. The timing of the oscillations was a function of the duration of the mechanical pulse. Furthermore, interaction was additive when the interval between stimuli was an even multiple of the pulse width. This resulted in a minimal point on the recovery curve. At odd multiples of the pulse width, the converse resulted. The evidence suggests that waves, set up in the skin by the paired mechanical pulses, interact and cause the recovery curve oscillations. Such waves could be set up by a variety of stimuli that contact the skin naturally.


Area 18 cell responses in cat during reversible inactivation of area 17. 1. A region of area 17 in the cat was temporarily inactivated by cooling, and cells in a topographically corresponding part of area 18 were studied during the blockage of area 17 input. 2. The responses of most area 18 cells were markedly reduced by cooling area 17 and recovered on rewarming primary visual cortex. 3. The selectivity of area 18 cells for oriented stimuli was not affected by inactivating area 17. These cells also, in general, retained their selectivities for the direction of stimulus movement and for the sign of contrast (light or dark) of a moving edge in the absence of input from area 17. 4. There was a greater decrease, on the average, of area 18 cell responses to slowly moving stimuli than to rapidly moving stimuli when area 17 was cooled; however, the exact pattern of change varied considerably from cell to cell. 5. These results suggest that cells in area 18 are to some degree dependent on input from area 17 for their responses, but to a large extent are independent in their selectivity for those stimulus parameters (orientation, direction of movement, sign of contrast) studied.


Nonequivalent visual, auditory, and somatic corticotectal influences in cat. 1. The effects of cortical cooling on the responses of cells to visual, somatic, and acoustic stimuli were studied in the cat superior colliculus (SC). When the visual cortex was cooled, the responses of many visual cells of the SC were depressed or eliminated, but the activity of nonvisual cells remained unchanged. This response depression was found in visual cells located in both superficial and deep laminae and was most pronounced in neurons which were binocular and directionally selective. 2. Cooling somatic and/or auditory cortex had no effect on visual SC cells and, with few exceptions, did not alter the activity of somatic or acoustic cells either. 3. The specificity of visual cortex influences on visual responding in the SC was most apparent in multimodal cells. In trimodal cells, the simultaneous cooling of visual, somatic, and auditory cortex eliminated responses to visual stimuli, but did not affect responses to somatic or acoustic stimuli. Visual responses were returned to the precooling level in both unimodal and multimodal cells by cortical rewarming. 4. The present experiments indicate that despite the organizational parallels among visual, somatic, and acoustic cells of the cat SC, the influences they receive from cortex are non-equivalent. Cortical influences appear to play a more critical role in the responses of visual cells than in the responses of somatic and acoustic cells. These observations raise questions about the functional significance of nonvisual corticotectal systems.


Segmental reflexes mediated by joint afferent neurons in cat knee. 1. Reflex effects mediated by knee joint afferent neurons were tested using the electrically evoked monosynaptic reflex in motoneurons innervating muscles which act about the knee. 2. Joint afferents were excited using small extension movements of the knee, stimuli which have been shown to be effective in activating a large fraction of the myelinated fibers in joint nerves. Because of the variability of reflex responses observed by others under similar experimental conditions, experimental control and data collection were done automatically under computer control. Large numbers of trials were presented in random sequence, and the resulting data were sorted and averaged. 3. Positive feedback was observed in both knee extensor (vasti) and knee flexor (posterior biceps-semitendinosus (PB-ST)) motoneuron populations. Vasti motoneurons were excited, and PB-ST motoneurons were inhibited by knee extensions when the posterior articular nerve (PAN) was intact; these effects were lost or substantially reduced when the PAN was cut. 4. Using knee extension stimuli, no reliable reflex effects were observed which could be attributed to afferents in medial articular nerve.


Organization of peripheral nerves and spinal roots of the Atlantic stingray, Dasyatis sabina. 1. The sizes and numbers of axons in peripheral nerves and spinal roots were investigated in the stingray, Dasyatis sabina. 2. The axons of the dorsal and ventral roots do not mingle in peripheral nerves of this animal as they do in higher vertebrates. Thus, it was usually possible to split the peripheral nerve into two portions, one containing only dorsal root axons, the other containing only ventral root axons. This feature was useful for the analysis of certain aspects of spinal cord organization. 3. The fact that dorsal and ventral root axons were segregated in peripheral nerves enabled us to demonstrate, without experimental surgery, that the central processes of the dorsal root ganglion cells and the proximal ventral root axons were 10-20% narrower, on the average, than the distal processes of the same dorsal root ganglion cells or the distal parts of the same ventral root axons. 4. The stingray is remarkable in having very few unmyelinated axons in the dorsal roots, ventral roots, or peripheral nerves. This paucity of unmyelinated axons distinguishes the Atlantic stingrays from all other vertebrates whose roots and nerves have been examined for unmyelinated fibers. 5. Similar findings were obtained for one spotted eagle ray (Aetobatus narinari) and two cow-nose rays (Rhinoptera bonasus).


Permissible concentration in air of xenon-127: concise communication. This paper reviews the method of calculation, and the criteria involved, in the determination of the maximum permissible concentrations of inert gases in ambient air. The results show that because the original calculations for Xe-133 included both photon and particulate dose, the permissible levels for Xe-127 are only slightly less than the levels established for Xe-133.


Left-ventricular ejection fraction and segmental wall motion by peripheral first-pass radionuclide angiography. A computerized edge-detection method was developed to obtain radionuclide ventriculograms for analysis of left-ventricular ejection fraction and segmental wall motion from first-pass studies following i.v. injection of radionuclide. The accuracy of this technique was examined in 21 patients undergoing cardiac catheterization. Tc-99m DTPA was injected into an antecubital vein, with data acquisition in the 30 degrees RAO projection by a gamma scintillation camera interfaced to a computer. A computerized profile analysis was used to determine objectively the edge of the left-ventricular blood pool. Time-activity curves were generated, and the ejection fraction was calculated from sequential end-diastolic and end-systolic count rates. The values for ejection fraction correlated well with those obtained by single-plane contrast ventriculography (r = 0.95). End-diastolic and end-systolic images were reconstructed from the time-activity curve. To analyze segmental wall motion, the left-ventricular outline was divided into five segments and the motion of each segment was graded qualitatively from 1 to 5. Seventy-five of 105 segments had the same grade as the wall motion determined by contrast angiography, and 102 of 105 were within one grade. (P less than 0.001). These findings demonstrate the accuracy of this improved technique for objective, rapid, and noninvasive determination of left-ventricular function.


Tc-99m pyridoxylidene glutamate in jaundiced patients. Ninety patients, 85 of them jaundiced, were examined after the injection of Tc-99m pyridoxylidene glutamate, a substance rapidly concentrated by normal hepatocytes and excreted into the biliary tract. It appears in the gallbladder after 10-15 min, and in the gastrointestinal tract at 30 min. On the basis of the time of appearance in the intestine, four groups of patients were recognized: 1. Seventeen patients with a normal pattern, with visualization of the gut at 30 min. 2. Nineteen patients with a slight delay in passage, with appearance in the gut between 30-180 min. 3. Twenty-nine patients showing very slow excretion visualized only at 24 hr; 21 of these had parenchymatous disease of the liver, three choledocholithiasis and five had malignant disease causing partial obstruction. 4. Twenty-five patients with no visualization of the intestine; eight of them proved to be cases of medical jaundice, and 17 were surgical cases. Groups 3 and 4 comprise 54 patients where the question of medical or surgical jaundice was critical. If lack of intestinal activity is considered an indicating surgical jaundice, the accuracy of this study was only 72.4%.


Accumulation and localization of gallium-67 in various types of primary lung carcinoma. The uptake and location of Ga-67 were investigated in 15 primary pulmonary carcinomas. The accumulation in the tumor was determined by scintigraphy of the patient, grain counts over fields of tumor cells in autoradiographs of tumor-tissue samples, and gamma counts in specimens of the tumor. Good correlation was found between the results obtained with these three methods. The relationship between accumulation of Ga-67 in the tumor and the histologic type of tumor was also studied. Undifferentiated carcinomas, and tumor cells in squamous-cell carcinomas showed significantly more Ga-67 than tumor cells in adenocarcinomas. No correlation was found between the presence of inflammatory infiltrates in or around the tumor and the grade of the scintigraphic images. In the autoradiograms, lymphocytes, plasma cells, granulocytes, and macrophages showed less radioactivity than the tumor cells--or none at all. Collagen fibers appeared to have bound some Ga-67, but necrotic areas showed no uptake.


Scintigraphy in the followup of pediatric splenic trauma treated without surgery. Twenty patients with blunt abdominal trauma were diagnosed as having splenic rupture by Tc-99m sulfur colloid scintigraphy. Because of the increased risk of infection in children following splenectomy, surgery was not performed and scintigraphic followup of the injured spleens was carried out. Thirteen patients were scanned at least 2 mo after trauma and the longest followup was 1 yr. Only three spleens showed "scintigraphic healing." The remaining ten showed smaller defects, but in two patients the size of the defect did not change after the 2-mo scan. In no case did the scan defect enlarge. Technetium-99m sulfur colloid scintigraphy offers a convenient and sensitive method of following trauma cases if no surgery is performed. Scintigraphic defects can persist for long periods of time while the patient is asymptomatic.


Studies on gallium accumulation in inflammatory lesions: II. Uptake by Staphylococcus aureus: concise communication. Gallium-67 was demonstrated to be taken up in significant amounts by a number of common micro-organisms. The mechanism of gallium-67 uptake was studied in S. aureus. It is found to involve two separate processes. One is insensitive to temperature or metabolic inhibitors, and is not inhibited by a high concentration of nonradioactive gallium. This process probably operates through nonspecific binding of Ga-67 to components of S. aureus. The second process is not inhibited by metabolic inhibitors either, but it is temperature-sensitive and is inhibited by high concentrations of stable gallium. This component of the Ga-67 uptake is most likely due to a carrier-mediated transport system (facilitated transfer).


A model for regional cerebral oxygen distribution during continuous inhalation of 15O2, C15O, and C15O2. Equilibrium positron brain scans were obtained during continuous inhalation of C15O, C15O2, and 15O2. Inhalation of C15O labels hemoglobin, whereas C15O2 instantaneously labels stable water to H215O. During the continuous inhalation of 15O2, body tissues extract it from the blood in proportion to local metabolism and ultimately convert it to water of metabolism. After 6-8 min of inhalation, a steady-state equilibrium is established in which the inflow of tracer is balanced by its disappearance due to radioactive decay (T1/2 = 2 min) and biologic removal. Mathematical models of the steady-state distributions of C15O, C15O2/H215O, and 15O/H215O are derived. The major results are: a) The steady-state distribution of C15O is insensitive to variations in blood flow and essentially measures blood volume. b) The distribution of H215O during inhalation of C15O2 is dependent, though nonlinearly, on blood flow. c) The distribution of H215O during inhalation of 15O2 depends linearly on the oxygen extraction fraction and nonlinearly on blood flow. d) The dependence on blood flow in the 15O2 steady-state image can be removed by the division, point by point, of the 15O2 image by the C15O2 image.


Problems in the interpretation of the in vivo measurement of calcium by the argon-37 method: an investigation of inert-gas elimination in humans. An investigation has been made of some physiological problems associated with the interpretation of in-vivo measurements of calcium by the argon-37 method. Inert-gas elimination in humans over a period of several days was studied using i.v. injections of Xe-133. The results imply that the exhalation rate of A-37 formed in bone will be affected by the individual's composition, in particular body fat. Comparison of calcium measurements between individuals and between laboratories is meaningful only if corrections are made for differing individual composition.


Liquid scintillation vial for radiometric assay of lymphocyte carbohydrate metabolism in response to mitogens. We have demonstrated that mitogens--i.e., PHA and Con.A--stimulate lymphocyte carbohydrate metabolism using a liquid-scintillation vial with conventional liquid-scintillation detectors. The results showed that this enclosed system can be useful for development of rapid in vitro tests of lymphocytes immune responsiveness, as well as for radiometric detection of bacterial growth in various gaseous atmospheres.


Detection of lipoid tumors by xenon-133. Three patients with biopsy-proven liposarcoma were studied with inhalation of xenon-133, a gas highly soluble in fat. Increased concentration of radioactivity in the region of the tumor suggested the potential usefulness of radioxenon for the detection of lipomatous tumors.


The monitoring of renal dysfunction in renal emphysema by dual radiopharmaceutical scintiscanning. Followup renal scintiscans using two radiopharmaceuticals were performed in a patient with renal emphysema. This allowed us to more accurately determine whether the emphysematous pyelonephritis was responding to antibiotic therapy or whether surgical intervention was indicated. Both Ga-67 citrate and a cortical agent (Tc-99m dimethylsuccinate) were used. A reciprocal relationship in the renal uptake of these two agents indicates the response of therapy and the degree of improvement of renal cortical function.


Imaging the bowel with technetium--an aid in gallium studies. Three cases are present to demonstrate the usefulness of oral or rectal technetium-99m preparations in locating the stomach and bowel in relation to abnormal accumulation of gallium-67 within the abdomen. In this way, the concentration of gallium in an abscess or a tumor may be distinguished from its physiologic excretion into the bowel.


[1-11C] DL-valine, a potential pancreas-imaging agent. There is a great need for a better pancreas-imaging agent. Studies with [1-14C] DL-valine have shown this amino acid to have a high pancreatic specificity in the four animal species examined. The tissue distribution was almost optimal by 30 min after injection, and no carrier effect was observed through a dose of 5 mg/kg. [1-11C] DL-Valine was synthesized in amounts up to 363 mCi using a rapid (T1/2 = 20.4 min for C-11), high-temperature, high-pressure modification of the Bücherer-Strecker amino acid synthesis. Purification was by anion-exchange followed by cation-exchange chromatography. [1-11C] DL-Valine was obtained in a 70% chemical yield with a total synthesis and purification time of 45 min. Studies in animals have demonstrated that it is a potentially useful new agent for clinical pancreatic imaging.


Biodistribution of intravenously injected [14C] doxorubicin and [14C] daunorubicin in mice: concise communication. [14C] doxorubicin (adriamycin) and [14C] daunorubicin (daunomycin) are cardiotoxic antibodies used in cancer therapy. These drugs were examined as possible agents for the measurement of regional myocardial blood flow. The antibiotics were injected intravenously into mice, which were then killed after various intervals. At a chemical dosage of 0.5 mg per kilogram, the content of the heart never exceeded 0.60% of the administered dose for doxorubicin and 0.55% for daunorubicin. The cardiotoxic effect of these drugs, therefore, is probably related to a specific sensitivity of the heart, rather than to an avid uptake of the drugs by the cardiac muscle. Further studies seem warranted, using a lower chemical dosage and higher specific activity.


Minimum detectable gray-scale differences in nuclear medicine images. An analysis of the effects of display-system characteristics upon the image from a nuclear medicine imaging system is presented. Simple, quantitative relationships are developed for calculation of contrast values for a minimum detectable lesion and maximum useful image count densities, from the slope of the D-log E characteristic curves of different films. The results provide a practical aid in the selection of films for various nuclear medicine imaging procedures.


Chest tomography by gamma camera and external gamma source: concise communication. To obtain tomographic images of the chest, we used a large-field gamma camera to detect the 90 degrees scattered radiations (180 keV) from a linear source of Hg-203 (279 keV). The primary beam traveling across the chest is scattered according to the relative density of tissues. Chest sections can be visualized at different depths on frontal and sagittal planes. The resolution of the technic is that of the gamma camera.


Effects of experimental diabetes and food intake on rat intestine and serum alkaline phosphatase. Alkaline phosphatase activity of rat serum was reduced 50% by fasting the animal for 24 hours. Diabetes, induced by alloxan or streptozotocin, increased serum alkaline phosphatase 3- to 5-fold in fed rats and the elevated activity was reduced by insulin administration. In the absence of insulin, fasting alone was able to reduce the serum alkaline phosphatase of diabetic rats to control values. The elevated serum isozyme was found to be of intestinal origin by the use of appropriate inhibitors and electrophoretic mobility following neuraminidase treatment. It is concluded that food intake, particularly the hyperphagia of diabetes, plays a major role in regulating the concentration of intestine and serum alkaline phosphatase in the rat.


Arginine requirements in immature dogs.

Arginine requirements in immature dogs. Labrador Retriever puppies (3 kg) were fed L-amino acid (L-AA) diets, containing the equivalent of 14% protein, to determine dietary argnine requirements for optimal growth and maintenance of normal intermediary metabolism. Growth and food consumption were depressed by decreasing the dietary arginine concentration. Urinary citrate and orotate increased with decreasing dietary arginine. Elevated blood orotate, urea and NH4+-N were detected in arginine deficient dogs. More than 0.56% arginine was required to support optimum growth and prevent abnormal loss of urinary metabolites. The effect of dietary nitrogen concentration (14, 21, or 28% L-AA) on arginine requirements was examined in immature Beagles. All arginine deficient dogs and dogs fed the 28% L-AA with arginine showed signs of emesis, excessive salivation and muscle tremors. Hyperammonemia and hyperglycemia were observed 2 hours after force feeding an L-AA diet devoid of arginine. Only hyperammonemia was observed in the Labrador Retrievers fed the same diet but incorporated into a 2% agar gel. Dietary nitrogen concentration or dietary arginine content dit not significantly influence glucose tolerance response to oral glucose loading. These data show that dietary arginine is required in the immature dog and that the requirement is influenced by dietary nitrogen concentration.


Effect of salicylamide on the placental transfer and fetal tissue distribution of sodium-35S-sulfate in the rat. Pregnant rats were used to determine effects of salicylamide on the sequential uptake and loss of radiosulfate by maternal and fetal tissues. Separate and combined effects of salicylamide and protein restrictions on radiosulfate retention by fetal rat tissues were determined. Rats were fed salicylamide-containing or control diets from the 4th or 6th day of gestation. Rats were killed between the 17th and 19th days of gestation, following intramuscular injection of sodium-35S-sulfate. Fetal and placental radiosulfate uptake was related to maternal serum levels. Salicylamide administration decreased radiosulfate uptake by maternal serum and liver, fetus and placenta--effects being dose-dependent. Differences in radiosulfate uptake by the fetus and placenta over time, induced by salicylamide, were also significant independently of maternal serum levels of radiosulfate. Major retention of 35S was found in the fetal cartilage with lower concentrations in the fetal skin, intestine, brain and liver. Protein restriction increased retention of 35S, and salicylamide administration caused a significant reduction in 35S retention by fetal tissues. Skeletal malformations were found in fetuses from dams receiving salicylamide. It is concluded that this drug lowers the availability of sulfate to the fetus from the dam, impairs the incorporation of sulfate by fetal tissues utilizing sulfates, and is teratogenic.


Oxidation, excretion, and tissue distribution of [26-14C] cholesterol in copper-deficient rats. The effect of copper deficiency on in vivo catabolism and excretion of [26-14C] cholesterol was studied in male rats. The study involved four treatments, namely, control, copper-deficient, control plus cholesterol, and copper-deficient plus cholesterol supplement. Significant elevations of serum ester and total cholesterol concentrations and reductions of serum free, ester, and total cholesterol specific activities were observed in rats fed the copper-deficient diets. In addition, a significant reduction of liver free cholesterol concentration was observed in rats fed the copper-deficient diets. Cholesterol supplementation significantly increased the concentrations of, and reduced the specific activities of, the different serum and liver cholesterol fractions. The only exception was that the liver free cholesterol concentration was not altered by cholesterol supplementation. The serum free cholesterol concentration was significantly increased and the specific activities of liver ester cholesterol were significantly reduced in rats fed the copper-deficient diet with no added cholesterol. The rates of oxidation and excretion of [26-14C] cholesterol were not influenced by dietary copper but were significantly increased by cholesterol supplementation. A shift of cholesterol from the liver to the serum pool appeared to be responsible for the hypercholesterolemia observed in copper deficiency.


Effects of age, sex and starvation on the turnover of plasma, liver and breast muscle proteins in the Japanese quail. Half-lives of the plasma, liver and breast muscle proteins in female, old male, young male and starved young male Japanese quail were measured by following the changes in protein specific radioactivity over a period of 7 days. The female birds showed the shortest half-lives for plasma and liver proteins and very long half-lives for breast muscle proteins. Compared to the proteins from the female birds, the old and young male birds showed slightly longer half-lives for plasma and insoluble liver proteins and shorter half-lives for soluble liver and breast muscle proteins. Starved birds showed greatly increased half-lives for all proteins when compared to the corresponding proteins from the other groups of quail. In general, the trends shown for the turnover of proteins from plasma, liver and breast muscle of the four groups of quail were similar to turnovers reported for rats under similar treatment conditions.


Studies on the role of nickel in the ruminant. Lambs were fed 6 to 7% of metabolic body weight per day of a basal purified diet low in nickel (65 ppb) or the basal diet plus 5 ppm nickel for a 97 day period in an attempt to demonstrate an essential role for nickel in the ovine. Weight gains for the entire period and digestibility of dry matter and of protein at 28 and 56 days were not different between the two groups. At 28 days, but not at 56 days, urinary nitrogen was less and percentage retention of absorbed nitrogen was greater in the supplemented lambs. Total serum proteins were higher at 56 days and serum alanine transaminase was higher throughout the experiment in the nickel supplemented lambs, but only significantly so at 56 days. When lambs were given an oral dose of 65Ni, the low nickel lambs tended to excrete more in the feces and retained less in the kidney, lung, and liver at 72 hours post dosing. The major excretory route of nickel was via the feces. The kidney retained the highest concentration of 65Ni of the organs examined.


Alterations of rat adipose tissue metabolism associated with dietary chromium supplementation. Net free fatty acid release from adipose tissue taken from male rats fed a stock diet unsupplemented with chromium was significantly greater than that from adipose tissue taken from chromium-supplemented rats. This enhanced net lipolysis was unaccompanied by any evidence of a compensatory increase in reesterification. Specifically, the glycerokinase-catalyzed direct phosphorylation of glycerol, which might have provided an alternate supply of alpha-glycerophosphate for reesterification in the absence of glycolytic production of alpha-glycerophosphate, was significantly reduced. These animals also showed an abnormal glucose tolerance response. In contrast, male rats fed a purified diet unsupplemented with chromium appeared to be marginally chromium-deficient as judged by a glucose tolerance test but did not show any alterations in adipose tissue metabolism attributable to the adequacy of chromium nutriture.


The influence of two barrier creams on the percutaneous absorption of m-xylene in man. Human volunteers were exposed to m-xylene by the percutaneous route (both hands immersed into the solvent for 20 minutes). The total amount of unchanged m-xylene eliminated through the respiratory tract during and after exposure and the total quantity of m-methylhippuric acid eliminated in urine during the subsequent 24 hours were measured. The majority of absorbed m-xylene (86 to 98%) is eliminated as m-methylhippuric acid in urine. The results obtained on eight subjects who repeated the test twice indicate that under similar experimental conditions the quantity of m-xylene absorbed percutaneously by an individual may vary with time by a factor of 2 or less. The interindividual variability in the total amount of m-xylene absorbed through the skin is greater than the intraindividual variability. Among 13 subjects, the ratio between the highest and the lowest value is 6.2; the mean percutaneous absorption rate of m-xylene amounts to 2.45 microgram/cm2 x min. The application of a barrier cream containing 10% silicone did not significantly influence the rate of percutaneous absorpiton of m-xylene by 11 subjects. Preliminary results obtained of five volunteers suggest that the application of a cream containing glycerol, stearates and oleostearates was not more effective.


Mortality of workers exposed to chloroprene. A study was undertaken to determine whether exposure to chloroprene increases the risk of lung cancer. Data were obtained from historical prospective mortality studies of two cohorts, one consisting of 270 men first exposed between 1931 and 1948, and the other of 1,576 men first exposed between 1942 and 1957. The number of lung cancer deaths in each cohort (three in the first and 16 in the second) were about the same as expected. Among maintenance mechanics in the second cohort, there were eight lung cancer cases (four living and four dead). A crude morbidity analysis suggested that this group may have had an excess incidence of lung cancer. However, the absence of excess lung cancer mortality in other high exposure occupational groups indicates that chloroprene exposure does not increase the risk of lung cancer.


Mortality of California agricultural workers. The mortality of white male farm laborers and farm operators in California for the years 1959 to 1961 was examined. Cause-specific mortality rates were calculated in order to identify diseases which might be of occupational importance. The mortality rate from respiratory diseases in the farm laborer groups (employees) was triple the rate in the farm management group (employers or self-employed individuals). Accidental deaths were also increased in the farm laborer group. Hazards of the agricultural industry which could contribute to accidents and respiratory disease were reviewed.


Differences in pulmonary function tests among whites, blacks, and American Indians in a textile company. Normal standards for pulmonary function in nonwhite populations are not presently available to occupational health workers. The present study examined differences in %FVC, FVC, FEV1/FVC, FEF 200-1200 and FEF 25-75% among whites, blacks, and American Indians. The sample consisted of 4209 job applicants to a textile company in southeastern United States. Multiple regression and analyses of covariance were employed to control differences in age, height, weight, and smoking status. Blacks were significantly lower in five of the six comparisons but higher in FEV1/FVC%. The pulmonary function measures of Indians fell generally between those of whites and blacks. Differences were observed between blacks and whites of both sexes regarding the effects of cigarette smoking. The need for race specific stardards was confirmed and the question of using smokers in the acquisition of normative data was raised.


Physiological effects on word performance of vapor-barrier clothing and full-face respirator. Work performance with impervious clothing and full-face respirator was investigated during maximal and submaximal (40-60%) bicycle ergometer efforts. Such levels may be dictated by the requirements of emergency decontamination tasks while most studies have been concerned with lower levels of activity. Five different exercise protocols were administered. Heart rate (HR), oxygen consumption (Vo2), skin (Ts) and rectal (Tr) temperatures were monitored. The impervious suits resulted in decreased work performance, aerobic metabolism, and tolerance time. Stress indices of HR and mean Ts were found to be correlated with these differences. The study is being expanded for the purpose of establishing safe physiological guidelines for work tolerances at higher energy levels in impermeable suits.


Localization of the site of urinary tract infections by means of antibody-coated bacteria in the urinary sediments. A prospective study was conducted in children with urinary tract infections to examine the relationship between the presence of antibody-coated bacteria in the urinary sediments and the presence of pyelonephritis. The site of the urinary tract infection was localized as upper or lower urinary tract by the bladder washout technique. Antibody-coated bacteria in the urinary sediments were identified by an immunofluorescent method. In 4 of 12 children with upper urinary tract infections, less than 1% of the bacteria in the urinary sediments were antibody coated. In 10 of 35 children with lower urinary tract infections more than 50% of the bacteria in the urinary sediments were antibody coated. These studies show no significant correlation between the presence of antibody-coated bacteria in urinary sediments and infection of the upper urinary tract.


Hepatitis B in children. II. Study of children born to chronic HBsAg carrier mothers. A survey of 4,452 pregnant women taken to find hepatitis B surface antigen revealed 28 asymptomatic chronic carriers. At birth, 16 of 17 infants studied were negative for HBsAg, whereas anti-HBc was present in all patients at a titer similar to that of the mother. Twelve children were followed for 6 to 18 months. In four of them, HBsAg remained negative and anti-HBc titers progressively decreased and were undetectable when tested after the age of 6 months. In eight infants, HBsAg became positive after an average time of 48 days. Elimination of HBsAg occurred in seven infants; five of them had clinical and biological manifestations of mild hepatitis. Sequential determinations of total complement and C components in three patients of the latter group showed dperession of complement at the time of appearance of clinical manifestations. In the patient who became an HBsAg carrier as well as in three infants who remained HBsAg negative, no decrease in complement titers was observed. These results indicate that vertical transmission from carrier mothers can occur in a low prevalence area and that neonatally infected children are capable of active elimination of HBV.


Cholelithiasis and Wilson disease.

Cholelithiasis and Wilson disease. We have studied three children with Wilson disease who had clinical symptoms suggestive of cholecystitis as well as radiologic evidence of gallstones, subsequently proven at the time of laparotomy. The gallstones from the patients with Wilson disease had an appreciably higher content of cholesterol than gallstones from age-matched children with hemolytic disease. Since gallstones may be present for years before they produce clinical symptoms, young patients with Wilson disease should undergo routine investigation for the presence of stones; cholelithiasis should also be considered in the differential diagnosis of abdominal pain in such patients.


Long-term pulmonary sequelae of the Wilson-Mikity syndrome. Five children born prematurely, eight to ten years ago, who survived the Wilson-Mikity syndrome were compared with six apparently normal prematurely born children and eight normal children born at term, by means of flow volume curves obtained while breathing air and while breathing a mixture of 80% helium and 20% oxygen. The flow rates in air of the group who survived the Wilson-Mikity syndrome were significantly lower than those of the prematurely born normal children (P less than 0.01), and than those of the eight normal children (P less than 0.001). The volume of isoflow while breathing air compared to that while breathing HeO2 was highly variable, but not significantly different in the three groups. Three of the five survivors of the Wilson-Mikity syndrome had a marked concavity in the shape of their flow volume curves. It is concluded that these abnormalities are due to focal increases of compliance or resistances or both of terminal lung units and result in adjacent areas of the lung emptying at different rates. The relationship between physiologic and anatomic abnormalities seen at autopsy in those who die to the disease suggests that they are due to the same pathologic process. Whether or not these abnormalities will disappear with future lung growth remains to be seen.


Modification of a screening program for neonatal hypothyroidism. From our experience in the screening of 212,000 newborn infants, we have devised a flow chart for processing T4 and TSH measurements obtained from initial filter paper blood spots. To date, all infants with thyroid dysfunction or TBG deficiency have been detected. Of the population screened, 1.84% require a spot TSH determination, and 1.1% require repeat determinations of T4.


Chlorambucil dosage in frequently relapsing nephrotic syndrome: a controlled clinical trial. A controlled clinical trial was performed using two dosage regimens of chlorambucil to treat children with frequently relapsing nephrotic syndrome. All children concurrently received prednisone (60 mg/m2 on alternate days). Ten children (Group I) were given chlorambucil as a stable dose (0.2 mg/kg/day) for 56 to 60 days, and 11 children (Group II) received increasing doses (0.2 to 0.63 mg/kg/day) for 42 to 77 days. Two children in each group subsequently relapsed. Follow-up averaged 28.6 and 27.2 months in Groups I and II, respectively. Three children in Group II developed infectious complications. The data indicate that a stable dosage regimen for chlorambucil is as effective as an increasing dose regimen in achieving long-term remission of frequently relapsing nephrotic syndrome.


Gonadal effects of chlorambucil given to prepubertal and pubertal boys for nephrotic syndrome. To assess the gonadal effects of chlorambucil given to prepubertal and pubertal boys, testicular function of 21 adolescents or young men, treated with chlorambucil for nephrotic syndrome before or during puberty, have been evaluated. Evaluations have been based on penis and testicular size, serum FSH, LH and testosterone values, and sperm count. Nine patients have testicular hypotrophy, 13 have significantly elevated FSH, 17 have azoospermia, two have severe oligospermia, and repetitive sperm counts show no improvement. A quiescent state of the epithelium of the seminiferous tubules does not protect against chlorambucil toxicity, which seems to be related to the cumulative dose. Chlorambucil should be used with caution, even in children.


Pattern of response to prednisone in idiopathic, minimal lesion nephrotic syndrome as a criterion in selecting patients for cyclophosphamide therapy. Twenty-three children with idiopathic, relapsing minimal lesion nephrotic syndrome were divided according to their pattern of response to prednisone: (1) steroid dependent, if the relapse occurred while the dosage of prednisone was being decreased; and (2) frequent relapser, if the relapse occurred at variable periods of time (one week to two months) after discontinuing prednisone therapy. All patients received cyclophosphamide for eight weeks in a single daily dose of 2 mg/kg, in order to prolong the length of the remission. The percentage of patients who continued in remission at the end of the first year and thereafter was greater in the frequent relapser group (P = 0.05). This study suggests that the pattern of response to prednisone may be another criterion for the selection of patients who will benefit from cyclophosphamide therapy.


The use of dopamine in children.

The use of dopamine in children. Dopamine was used as the primary catecholamine to treat circulatory shock, manifested by either systemic arterial hypotension or oliguria or both, in 24 children two days to 18 years (mean = 39 months) of age. The dose of dopamine ranged from 0.3 to 25 (mean = 9.3) microgram/kg/minute. The primary problem in four of the 24 patients was infection; two of these patients survived. The other 20 patients had congenital heart disease; 18 developed shock following surgery. Even of these 20 patients survived. With dopamine infusion the average systolic blood pressure increased from 69 +/- 4 (mean +/- SEM) to 81 +/- 4 mm Hg (P less than 0.001) and the mean urine output increased from 0.8 +/- 0.2 to 2.7 +/- 0.8 ml/kg/hour (P less than 0.05). Dopamine produced no adverse consequences. Thirteen patients responded favorably to the drug, with a significant increase in systemic arterial blood pressure and urine production. Four patients did not respond to dopamine and seven had an equivocal response. None of the four patients in whom dopamine was ineffective survived. Although only nine of the 20 patients who responded favorably or equivocally survived, conventional therapy had failed to alter the unfavoarble course in any of the patients.


Phenobarbital and diphenylhydantoin levels in neonates with seizures. Little information is available regarding appropriate plasma levels of anticonvulsant drugs in neonates. We determined the plasma levels of phenobarbital and diphenylhydantoin following initial administration and during maintenance therapy in 59 neonates with seizures. Following intravenous administration of 15 to 20 mg/kg, levels of 20.7 +/- 4.4 microgram/ml were achieved for phenobarbital and levels of 14.5 +/- 3 microgram/ml for DPH. Maintenance doses of 5 mg/kg of phenobarbital resulted in initial drug accumulation followed by more rapid elimination of the drug with increasing duration of exposure. Therapeutic plasma levels of DPH could not be achieved by oral administration in the neonate.


Affect, accessibility of material in memory, and behavior: a cognitive loop? Two studies investigated the effect of good mood on cognitive processes. In the first study, conducted in a shopping mall, a positive feeling state was induced by giving subjects a free gift, and good mood, thus induced, was found to improve subjects' evaluations of the performance and service records of products they owned. In the second study, in which affect was induced by having subjects win or lose a computer game in a laboratory setting, subjects who had won the game were found to be better able to recall positive material in memory. The results of the two studies are discussed in terms of the effect that feelings have on accessibility of cognitions. In addition, the nature of affect and the relationship between good mood and behavior (such as helping) are discussed in terms of this proposed cognitive process.


Stress effects on affiliation preferences among subjects possessing the type A coronary-prone behavior pattern. An experiment tested the hypothesis that when subjects who display the Type A coronary-prone behavior pattern are placed under stress, they prefer to wait for the stressful event with others, but desire to work under stress alone. One half of 25 Type A and 25 Type B (non-coronary-prone) subjects were told that they would receive painful electric shock while working on a mental task, while the other one half were told that they would receive subliminal stimulation. Both groups were then given the choice of waiting for the event with others or alone and the choice of working on the task alone, in the company of others, or in a leader-directed group. The results showed that Type As relative to Type Bs tended to wait in the company of others regardless of threat level, but displayed a marked preference to work alone under high threat. This decision to work alone was not influenced by waiting preference. Subsequent correlational studies showed that coronary patients (n = 40) reported a greater preference for working alone when under pressure than matched controls (n = 40), as did Type A college students (77% vs. 14% for Type Bs) when asked their preference within the context of the structured interview that is used to assess the Type A pattern. The significance of the findings for increasing the understanding of affiliative preferences and coronary-prone behavior are discussed.


Kinetics of plasma coagulation and lysis I: Basic kinetic model for time course of coagulation-lysis systems and its potential application to clinical studies. The time courses of coagulation and coagulation-lysis were spectrophotometrically monitored after the addition of thrombin or thrombin-streptokinase to plasma, diluted 1:5 with normal saline, obtained from normal and presumably abnormal subjects. The kinetics of clotting, after an initial lag period of 0.5-1.5 min, demonstrated essentially first-order dependence on the amount of fibrinogen available to form the clot, and the asymptotic absorbance was independent of thrombin concentration. The rate of clotting was a function of added thrombin, and the ratios of the rate constants at 2.5 and 1.25 units of thrombin/ml of undiluted plasma were 1.65 +/- 0.03 SEM. At early times, the coagulation-lysis curve with thrombin-streptokinase could be superimposed on the clotting curve with thrombin alone for a given plasma with minor compensation for variable lag times. Subsequently, the curves diverged; lysis was monitored by the decrease in absorbance of the coagulation-lysis system. The rate of fibrinolysis increased with streptokinase concentration and was a function of the extent of lysis, and it permitted the description of the kinetics of lysis by a pseudoautocatalytic mechanism where the bimolecular rate constant appears proportional to streptokinase concentration. Ranges of clotting and lytic parameters for the plasma of normal subjects are given, and their potential use in diagnosing abnormalities is described.


Plasma concentrations of thioridazine metabolites and ECG abnormalities. Five acute schizophrenic patients were given a standard thioridazine dose (4 mg/kg/day po). Their plasma concentrations of thioridazine and its metabolites were determined by GLC, and their ECG's were recorded routinely. In four of the five patients, abnormal ECG's (malformation of the T-wave or lengthening of the repolarization time) were found whenever plasma concentrations of the thioridazine ring sulfoxide were elevated. No relationship could be detected between the appearance of ECG abnormalities and the concentrations of thioridazine and its other metabolites.


Potential CNS antitumor agents-phenothiazines II: fluphenazine analogs. Fluphenazine was found to possess moderate reproducible activity against the intraperitoneal L-1210 and P-388 leukemia murine tumor models. Seven ether derivatives of fluphenazine and eight compounds in which the terminal side-chain hydroxyl group was replaced by an amine function were prepared and evaluated in the intraperitoneal L-1210, P-388, and B16 melanoma systems as well as the intracerebral L-1210 and ependymoblastoma brain tumor models. While no substantial intracerebral activity was observed, seven derivatives possessed reproducible activity in the intraperitoneal L-1210 or P-388 system. Several gave T/C values of 150%. No B16 melanoma activity was observed. These compounds were also tested for their cytotoxic properties in culture against L-1210, P-388, and KB cells. The amine isosteres, while possessing little in vivo activity, were the most cytotoxic of the compounds prepared, with several having ED50 values less than 1 microgram/ml.


Molecular complexing ability of quinoline and its simple derivatives. Electron donor-acceptor complexes for a group of quinolines and naphthalenes with 9-(dicyanomethylene)-2,4,7-trinitrofluorene in 1,2-dichloroethane were studied by optical absorption methods. Association constants, molar absorptivities, and charge-transfer transition energies were evaluated for each system, together with theoretically calcualted orbital energies and complex geometries. In contrast to the association constants and structures reported for N-heterocycle-halogen complexes, these studies indicate that, with a moderately large pi-electron acceptor, quinolines function as pi-rather then n-(lone-pair) donors. These results support intercalation models for drug-receptor interactions involving the quinoline moiety.


Alkaloids of Strychnos dolichothyrsa Gilg ex Onochie et Hepper. Three alkaloids of the stem bark of Strychnos dolichothyrsa Gilg ex Onochie et Hepper (Loganiaceae) were isolated and identified as caracurine V and its mono- and di-N-oxide by comparison with synthesized compounds. 13C-NMR was used to confirm the structure of the N-oxides. The muscle relaxant activity and the toxicity of the N-oxides were less than those of caracurine V.


Light-scattering studies on bile acid salts I: Pattern of self-association of sodium cholate, sodium glycocholate, and sodium taurocholate in aqueous electrolyte solutions. The pattern of association of the trihydroxy bile salts in aqueous electrolyte solutions was investigated utilizing the light-scattering technique. The turbidity of the bile salts sodium cholate, sodium taurocholate, and sodium glycocholate was determined over the concentration range of 0-25 mg/ml at 25 degrees. For sodium cholate, the concentration of the supporting electrolyte was varied from 0.15 to 0.5 M. For all bile salts in 0.15 M electrolyte, the turbidity was determined in sodium fluoride, sodium chloride, sodium bromide, and sodium iodide. Comparison of the light-scattering data with amonomer-micellar model showed that qualitative agreement was obtained; however, quantitative agreement could not be achieved. Further examination of the data showed that the light-scattering results were in good agreement with a model that includes dimers, trimers, and a higher aggregate containing approximately eight monomeric units.


Simple, rapid and micro high-pressure liquid chromatographic determination of endogenous "true" creatinine in plasma, serum, and urine. Previously published methods for endogenous creatinine levels in plasma, serum, or urine lack specificity or are subject to interferences from endogenous or exogenous substances. The developed simple, rapid, and specific high-pressure liquid chromatographic method includes the novel deproteinization and extraction of 1 volume of plasma or serum with 2.5 volumes of acetonitrile and also of 1 volume of urine with 40 volumes of a 20% water-80% acetonitrile solution. An aliquot of the supernate is then injected directly into the chromatograph. A cation-exchange column and acidified (0.02% of 85% phosphoric acid) 0.1 M ammonium phosphate solution as the mobile phase, with a flow rate of 2 ml/min, were used. Creatinine, with a retention time of 3.8 min, was monitored via its UV absorption at 215 nm. Both peak heigh and integrated area methods of quantitation yielded the same results. Several methods were employed to show that the "suspected" creatinine peak from plasma samples was due entirely to the "true" creatinine. No interference was found in samples obtained from normal and renal patients. The day-to-day variation in the detector response was small. Each assay requires only about 5 min for completion. Ten microliters of plasma or serum or 1 microliter of urine is sufficient for analysis.


General treatment of linear pharmacokinetics. A general treatment of linear pharmacokinetics that enables equations to be obtained simply for all linear compartmental models, with input in one or more compartments, is presented. Two approaches are described: one based on a full Laplace transformation and one that avoids transformation of the input functions and the use of convolution integrals. The latter approach is of particular interest when dealing with complex input functions not having a simple Laplace transform. The concept of acceptor and donor subsystems is introduced. It is demonstrated that disposition in certain models may be simplified and analyzed in terms of disposition in subsystems of simpler composition. The treatment presented is illustrated with several examples.


Physiological perfusion model for cephalosporin antibiotics I: Model selection based on blood drug concentrations. Various cephalosporins with different degrees of protein binding were administered to human volunteers. Blood samples were collected as a function of time and were assayed for drug content by a microbiological assay. A pharmacokinetic analysis of the data was performed using a two-compartment model with and without protein binding in the central compartment and a perfusion model. Both the two-compartment model without protein binding and the physiological perfusion model adequately described the blood levels of all three cephalosporins.


Stabilization of sulfisomidine tablets by use of film coating containing UV absorber: Protection of coloration and photolytic degradation from exaggerated light. The effect of model polymer coating films of vinyl acetate, containing oxybenzone as a UV absorber, on the coloration and photolytic degradation of simple sulfisomidine tablets was examined to attempt stabilization of photosensitive solid dosage forms. Coloration of the tablet surface was followed by the tristimulus colorimetric method in the fading tester equipped with a mercury vapor lamp. Photolytic degradation in the UV region was investigated by a new method for measuring the absorption spectra of a crystal sample in the gas phase, i.e., the semi-integral attenuance spectra. Two parameters of a film, thickness and concentration of the UV absorber, were varied at every exposure. These physical and chemical changes are discussed in relation to light transmission properties of films.


Electron-capture detector GLC technique for estimating tocainide in biological fluids. A sensitive and specific electron-capture detector GLC method capable of detecting picogram quantities of tocainide, a lidocaine analog, in biological fluids was developed. This method consists of extracting the compound into methylene chloride and derivatizing with heptafluorobutyric anhydride to form the monoheptafluorobutyryl derivative. The derivative formation was confirmed by GLC-mass spectrometry. Quantitative estimation was performed using 1-bromonaphthalene as an internal standard. The minimum detectable level by the electron-capture method was approximately 30 pg/injection as opposed to approximately 3 ng/injection with a flame-ionization detector Linear response was observed in the range from 50 pg to 3 ng using an electron-capture detector. No interference from endogenous substances was observed.


Bilirubin dynamics in the Gunn rat during phototherapy. Bilirubin dynamics were studied in homozygous Gunn rats under normal room lighting conditions and under conditions simulating phototherapy. A kinetic model was developed for the formation, distribution, and elimination of bilirubin. The decrease in plasma bilirubin concentration during illumination with low intensity [300 footcandles (fc)] and high intensity (1000-1100 fc) light was studied. The plasma bilirubin concentration in the rats decreased under phototherapy until a new steady-state concentration was reached, the decline being more rapid under high intensity light conditions. Gunn rats were also injected with a tracer dose of 14C-bilirubin following a period during which the rats were illuminated with low or high intensity light. The distribution and elimination of the labeled bilirubin were followed under continuous illumination. The chosen kinetic model, when adapted to the set of data under investigation, fit all of the data concerning bilirubin kinetics in Gunn rats under continuous illumination.


Effect of intensity of agitation on disintegration time of tablets. The effect of the intensity of agitation on the disintegration time of tablets was studied. Preliminary results obtained using tablets fabricated under controlled conditions and selected commercial tablets revealed that the intensity of agitation produced in the test apparatus was not reproducible with fixed geometry for successive runs when the settings were maintained constant. The disintegration times increased at a lower intensity of agitation. The intertablet range values obtained were rather large at the start and increased with a corresponding decrease in the intensity of agitation. The results showed that the measurement of disintegration times with the existing apparatus may not serve as a guide to the pharmaceutical formulator in the preparation of optimal dosage forms of drugs for clinical trial and may not ensure lot-to-lot uniformity of the pharmaceutical product.


Thermal degradation of gelatin as applied to processing of gel mass. The rheological parameters of rigidity and viscosity were studied for gelatin, both as a 6% solution and a 38% gel mass containing glycerin, sorbitol, and water. A compressive test for rigidity and a rotational shear method for viscosity were used to characterize the thermal degradation of several lots of alkali-processed calf-skin gelatin. The objective of this study was to use kinetic data for process predictions in the manufacturing of the gel mass for soft gelatin capsules. Empirical equations relating degradation to time, temperature and pH were derived for dilute gelatin solutions. Through parallel studies with a concentrated gel mass, other empirical equations were developed to predict degradation of the gel mass based on dilute solution data and processing conditions. Analysis of the kinetic data and empirical equations have generally confirmed earlier observations on degradation, except on quantitative aspects. While these equations are adequate for the intended use in process predictions, the study revealed substantial variability both within and among lots of commercial gelatin. These variabilities of uncertain causes contributed to inexact characterization of degradation, as demonstrated by the generally approximate results of process predictions.


Ascorbate stimulation of salicylate absorption in isolated rat intestine. Salicylate transport across the rat intestine was measured using both in vitro and in vivo procedures. After the steady state for labeled salicylate was reached, the addition of ascorbate stimulated tracer flux with the establishment of a new steady state. The tissue permeability had a saturable dependence on ascorbate concentration. Also, ascorbate stimulated the tissue short-circuit current.


Metabolic studies of the nonnutritive sweeteners cyclopentylmethylsulfamate and cyclopentylsulfamate: determination of metabolites in rat urine. The nonnutritive sweetener sodium cyclopentylmethyl-sulfamate was fed to Wistar albino rats. The urine was collected for 3 days, combined, and examined (GLC) for the metabolites cyclopentyl-methylamine and cyclopentylmethanol. The percent conversion to these metabolites was 0.077 and 0.0102, respectively. The percent conversion to these to cyclopentylmethylamine was the lowest conversion to amine observed when compared to the metabolism of three other sweet sulfamates, cyclopentylsulfamate, cycloheptylsulfamate, and cyclooctylsulfamate, previously administered to rats. The average excretion of unmetabolized sulfamate was 15.4%. Sodium cyclopentylsulfamate was fed to rats over 9 days, and an analysis was carried out for the metabolites cyclopentylamine, cyclopentanone, and cyclopentanol. A decrease in the level of metabolites occurred after the first 3 days.


Fenoprofen: drug form selection and preformulation stability studies. Several fenoprofen salts were prepared to obtain the most acceptable form for an oral dosage formulation. Thermal analysis techniques were used to compare stabilities of the water of hydration in different salt forms and to assess the effects of the water of hydration on compatibility with propoxyphene and codeine salts. Photodegradation products of fenoprofen were isolated and identified, and their relevance to product formulation was evaluated.


Analysis of the prodrug 7-acetylacroninium perchlorate in presence of the parent compound acronine. Analysis of acronine and the prodrug acetylacroninium perchlorate in parenteral solutions is described. The unstable prodrug reacts quantitatively with the nucleophilic agent aniline by an unusual mechanism to form a phenylimino derivative of acronine. This derivative and acronine itself were determined by a spectrophotometric two-component method.


GLC determination of phenylbutazone in human plasma. A GLC method for phenylbutazone at concentrations down to 10 ng/ml in human plasma is described. After addition of an internal standard, phenylbutazone is extracted at pH 5 into benzene. The dry extract is dissolved in benzene, and phenylbutazone is determined by GLC using a 63Ni-electron-capture detector.


Effect of enzyme-inducing and enzyme-inhibiting agents on drug absorption I: 3-O-methylglucose transport in rats. The influence of preadministration of phenobarbital, benzo[alpha]pyrene, and proadifen hydrochloride on 3-O-methylglucose transfer across the everted rat small intestine was examined. The active and passive components of the sugar transport mechanism were evaluated using phlorizin, a potent inhibitor of active transport of sugars. Pretreatment of the animals with phenobarbital did not alter 3-O-methylglucose transfer characteristics. Pretreatment with intraperitoneally administered benzo[alpha]pyrene increased mucosal to serosal transfer of the sugar low (0.1 mM) sugar concentrations. Enhancement of the active transfer of 3-O-methylglucose by pretreatment with proadifen hydrochloride was noted at low sugar concentrations. The passive transfer of the sugar was reduced after pretreatment with proadifen hydrochloride.


Plant anticancer agents V: new bisindole alkaloids from Tabernaemontana johnstonii stem bark. The isolation and structure elucidation of the three new bisindole alkaloids, gabunamine, tabernamine, and 19,20-epoxyconoduramine, from Tabernaemontana johnstonii stem bark are described. The isolation of the seven known alkaloids, conodurine, conoduramine, gabunine, isovacangine, ibogamine, pericyclivine, and perivine, from the same source also is noted. The alkaloids gabunamine, gabunine, and tabernamine showed significant cytotoxicity against the P-388 cell culture system.


Polynitro aromatic compounds in analytical chemistry II: Reaction of menadione with 2,4-dinitrophenylhydrazine. The intense blue color formed in the official assay of menadione injection by treatment of the sample with 2,4-dinitrophenylhydrazine and ammonia is shown by data from visible light spectra and mass spectra and by comparison of the pKa value with that of the corresponding reaction product of acetone to be due to proton abstraction from a monohydrazone.


Quantitation of etorphine in urine by selective ion monitoring using tritiated etorphine as an internal standard. Selective ion monitoring combined with GLC was used for the assay of etorphine in urine. Commercially available tritiated etorphine was added as an internal standard. The advantage of the methodology using this internal standard is higher sensitivity by a factor of about 20 when compared with ordinary GLC.


Drug partitioning: relationships between forward and reverse rate constants and partition coefficient. The rate constant, k1, of drug transport from an aqueous phase to an organic phase and the rate constant, k2, of the reverse process can be described as functions of the partition coefficient, P:logk1 = log P-log (betaP + 1) + c' and logk2 = -log (betaP + 1) + c'. In a homologous series, where log P is a simple function of the number of CH2 groups, log k1 and log k2 also can be described as functions of the number of CH2 groups. The relationships between these equations and current physicochemical models of drug absorption are discussed.


High-pressure liquid chromatographic determination of camphor and parachlorophenol in camphorated parachlorophenol. Quantitative analyses of both camphor and parachlorophenol in camphorated parachlorophenol by high-pressure liquid chromatography is described. A 5% solution of phenol in the mobile phase (heptane-chloroform 3:2) served as the internal standard; the assay involved addition of this phenol solution to the camphorated parachlorophenol, followed by further dilution using the mobile phase (heptane-chloroform, 3:2) and injection into the instrument.


Validity of oral bioavailability estimates of phenolsulfonphthalein based on total urinary excretion from rats. Urinary recovery of phenolsulfonphthalein from rats were determined after intracardial (0.075 mg) and oral (1.5 mg) doses. Although trace quantities of conjugated metabolites could be identified by TLC, the levels present did not introduce significant error into estimates of total phenolsulfonphthalein excretion if samples were assayed directly by colorimetric methods for only unchanged dye. The absolute availability of phenolsulfonphthalein based on urinary recovery under the present experimental conditions was estimated at 10.6%.


Rapid high-pressure liquid chromatographic determination of quinidine and dihydroquinidine in plasma samples. A high-pressure liquid chromatographic assay for quinidine and dihydroquinidine was developed. A cation-exchange column was utilized with an eluting solvent pH of about 9. The internal standard was cinchonine. The reproducibility and precision of this method were evaluated by analyzing replicate samples and by comparing results with those obtained from a TLC-fluorometric procedure. In addition, several drugs were evaluated to ascertain whether they interfered with the analysis of quinidine and dihydroquinidine.


Plant anticancer agents VII: Structural effects on cytotoxicity of bisindole alkaloids of voacamine type. Structural effects on the cytotoxicity of bisindole alkaloids of the voacamine series were investigated with compounds isolated from various Tabernaemontana species and compounds synthesized specifically for this purpose. Activity is sensitive both to the position of attachment of the vobasane unit on the iboga moiety and to the presence of an N-methyl group on the vobasane unit.


Differential borohydride assay for conjugated ketosteroids: improvement with propylene glycol. A previously developed differential UV assay for 3-one-4-ene steroids involving reduction with sodium borohydride was frequently found to be less than 100% complete. Such incomplete reduction can be mimicked by additions of sodium metaborate, a product of the hydrolysis of sodium borohydride, to the reaction system. Complete reduction can be achieved by adding propylene glycol to the reduction system. The phenomenon was studied using halcinonide as a model.


Quantitative structure-activity relationships and carminative activity. Carminative activities of 34 alcohols, esters, ethers, phenols, and carbonyl compounds were determined using the guinea pig isolated ileum preparation and are expressed as the ability to produce a 50% inhibition (ID50) of a standard response to carbachol. Aqueous solubilities were measured at 37 degrees using either UV absorption or GLC. The ratios of solubility to ID50 were reasonably constant, suggesting nonspecific biological activity, similar to that previously observed with general anesthetics. Hansch analysis indicated that carminative activities were largely controlled by solubility, as indicated by octanol-water distribution coefficients. The principal remaining factor appeared to be the steric availability of the oxygen atom in the functional group of the compound.


GLC determination of plasma concentrations of gamma-oxo metabolite of phenylbutazone. A quantitative method for the gamma-oxo metabolite of phenylbutazone from plasma is described. The procedure involved an ethylene dichloride extraction of acidified plasma to which an internal standard, acenocoumarol, had been added. The extracted gamma-oxo metabolite and the internal standard were methylated and analyzed by GLC. Determination of 0.25 microgram of gamma-oxo metabolite/ml with a relative standard deviation of 6.5% was accomplished.


Synthesis and antibacterial evaluation of 2-(substituted phenylureido)-4-thiocyanatobenzothiazoles. The synthesis and antibacterial evaluation of a number of 2-(substituted phenylureido)-4-thiocyanatobenzothiazoles are described. The more active compounds against the test organisms in vitro generally were those substituted with halogens on the phenyl and benzothiazole rings.


Total synthesis of 1,11- and 3,11-diazasteroids. The 2- and 6-methoxy derivatives of 8-aminoquinoline, as well as 5-aminoisoquinoline, condensed with 2-ethoxycarbonylcyclopentanone to give tricyclic intermediates in 86-90% yield. These secosteroids were cyclized in 35-69% yield with polyphosphoric acid to 2-methoxy-1,11-diaza-, 7-methoxy-1,11-diaza-, and 3,11-diaza-1,3,5,7,9,13-gonahexaen-12-ones, respectively. The latter exhibits slight antileukemic activity.


Structures of silver sulfonamides.

Structures of silver sulfonamides. The structures of silver sulfonamides were found to depend highly on the substituent at the amide nitrogen of the sulfonamide. Silver is coordinated to that nitrogen and the sulfonamide is in the amido form if no substituent is present or if the substituent is a phenyl, acetyl, or 2-pyrimidyl group. If the substituent is a 2-thiazolyl or 2-pyridinyl group, the sulfonamide is in the imido form and silver coordinates to the nitrogen of the substituent. Depending on the number of suitable donor atoms per sulfonamide, the silver compounds are charged or uncharged and the primary amino group may be involved  in complexation.


Effects of lidocaine and on slow response and depressed fast response action potentials of canine cardiac Purkinje fibers. Disease may decrease resting potential of cardiac fibers, thereby depressing the upstroke velocity of the action potential, causing slow conduction and reentry. A decrease in resting potential may also cause automaticity. We studied the effects of lidocaine (5 and 20 mg/l) on canine Purkinje fibers with reduced membrane potentials with either depressed Na+-dependent upstrokes (depressed fast responses) or with slow inward (Ca++) current-dependent upstrokes (slow responses). Depressed fast responses were produced by elevating [K+]0 in the perfusate, reducing membrane potential to around -60 mV, without abolishing excitability. Slow responses were produced by either perfusing fibers with a Na+-free, Ca++-rich solution, or by perfusing them with a high [K+]0 Tyrode's solution containing norepinephrine. Lidocaine had a marked depressant effect on depressed fast response action potentials. The drug markedly decreased Vmax and conduction velocity. It sometimes decreased action potential amplitude and caused conduction block. Resting potential was not changed. On the other hand, lidocaine had little effect on slow response action potentials. Resting potential, Vmax and action potential amplitude were not altered nor was conduction changed. The rate of spontaneous impulse initiation was slightly reduced by 5 mg/l of lidocaine but not by 20 mg/l. We conclude that lidocaine does not exert its antiarrhythmic effect by directly depressing the slow inward current but may be antiarrhythmic because it depresses an already depressed fast inward current and can cause conduction block.


Denervation-induced changes in electrophysiologic parameters of the smooth muscle of the guinea-pig and rat was deferens. In order to further elucidate the mechanisms by which postganglionic denervation causes changes in the dose-response curves obtained in smooth muscle, microelectrodes have been used to investigate cellular changes in the denervated guinea-pig and rat vas deferens. In the guinea-pig vas deferens, chronic denervation produced a partial depolarization (mean change of 8.5 mV) without any change in threshold for the action potential. In the rat vas deferens there was no change in resting potential but the threshold membrane potential because more negative (6.5 mV). Thus, in both species, but apparently by different mechanisms, the resting and threshold membrane potentials are brought closer together by denervation. Such an effect would clearly contribute to the well documented increase in sensitivity to depolarizing agonists which is produced by chronic denervation. In both species, denervation increased the space constant of the smooth muscle, an indication of increased electrical coupling among the cells. This observation is consistent with morphologic evidence of improved coupling induced by denervation and presented previously from this laboratory. The improved coupling appears to be associated with the increased maximum response of the denervated vasa deferentia of both species. These results are discussed in references to known similarities and differences in electrophysiologic characteristics between normal guinea-pig and rat vasa deferentia.


Potentiation by dipyridamole of the inhibition of guinea-pig ileum twitch response caused by adenine derivatives. The inhibition actions of adenosine and adenine nucleotides, such as ATP, AMP and cyclic AMP, on contractions of guinea-pig from ileum induced by transmural stimulation were potentiated by dipyridamole, whereas those of inosine, morphine and tetrodotoxin were not affected. Tritium activity, accumulated during incubation of the ileal segments with [3H]adenosine, was reduced by dipyridamole. The adenosine added was degraded to inosine and then hypoxanthine during incubation, with the ileal segments, thereby restoring the twitch response of the ileal segments. In the presence of dipyridamole, the degradation of adenosine and the recovery of the twitch response was retarded. Thus, dipyridamole may potentiate the inhibitory actions of adenosine and adenine nucleotides on the twitch response of the ileum by inhibiting both the accumulation and the degradation of adenosine.


Human placental cholinergic system: stimulation-secretion coupling for release of acetylcholine from isolated placental villus. Isolated villus from human term placenta contains about 167 nmol/g of acetylcholine (ACh). It was incubated in a muscle bath containing Krebs-Ringer bicarbonate buffer (pH 7.2-7.4) at 37 degrees C and ACh released into the medium was analyzed by pyrolysis-gas chromatography. The spontaneous release of ACh into the medium was linear with time and was 35 pmol/g/min. ACh was not released when Ca++ was absent in the medium. Raising the Ca++ concentration in the Krebs-Ringer bicarbonate buffer from 2.34 to 4.64 mM, or adding L-nicotine (58 muM) to the bath, increased the rate of release of ACh to 53 and 47 pmol/min respectively. Nicotine did not exhibit any effect on ACh release in the absence of Ca++ in the medium. Both the rate of the spontaneous release of ACh and the nicotine-induced increase in the release of ACh was decreased by atropine (152 muM). They were not influenced by d-tubocurarine (30 muM). Depolarizing concentrations of potassium (16-63 mM) in the medium increased the rate of release of ACh. Cocaine, a known Ca antagonist, decreased the rate of spontaneous release of ACh as well as nicotine-induced release of ACh. These observations indicate that 1) Ca++ ions in the external medium are required for release of ACh, 2) Ca++ ions in the external medium are required for release of ACh, 2) Ca++ ions act as a link between the stimulation of ACh release and the final release of ACh and 3) The effect of nicotine on placental release of ACh may be classified as a muscarinic type.


Effect of tolmetin on renal function and prostaglandin metabolism. The effect of tolmetin on prostaglandin synthesis by minces of rat renal medulla and on prostaglandin cyclooxygenase of rabbit renal medulla was determined in vitro. The effect of tolmetin was compared to the effects of indomethacin and ibuprofen. Pretreatment of rats in vivo with tolmetin, indmethacin or ibuprofen reduced prostaglandin synthesis by minces of renal medulla. Incubation of medullary tissue in medium containing tolmetin or indomethacin also decreased prostaglandin production. Both drugs reduced O2 consumption by prostaglandin cyclooxygenase from rabbit renal medulla. In addition, the effect of tolmetin, indomethacin and ibuprofen on renal blood flow and the intrarenal distribution of renal blood flow was measured in anesthetized dogs. Tolmetin and ibuprofen resemble indomethacin in reducing renal blood flow and in shifting the distribution of renal cortical flow from the inner cortex toward the outer cortex. It is concluded that tolmetin is an effective inhibitor of prostaglandin synthesis and affects renal function in a fashion similar to other prostaglandin synthesis inhibitors.


The development of the reproductive organs of the male giraffe, Giraffa camelopardalis. The reproductive organs of the male giraffe differ little in morphology and histology from those of other ungulates. There is some indication that gonadal hypertrophy occurs in late fetal life. Spermatogenesis begins at 3-4 years of age and coincides with a rapid increase of testicular weight and seminiferous tubule diameter. In the fetal testis the main hormone is androsteredione (2.73 microgram/g) but in adult testes testosterone is predominant (less than 10.08 microgram/g) and delta' testosterone may also be present.


Relationship between sperm concentration and the incidence of polyspermy in mouse embryos fertilized in vitro. A dose-response relationship between the incidence of polyspermy and sperm concentration was found by analysing chromosome preparations of first-cleavage mouse embryos derived from fertilizations in vitro with 3 concentrations of mouse spermatozoa (2 x 10(4), 2 x 10(6) and 1 x 10(7)/ml).


The effect of sperm and egg genotype on the incidence of chromosomal anomalies in mouse embryos fertilized in vitro. Analysis of the chromosomes of 1-cell mouse embryos fertilized in vitro showed that the incidence of polyploidy is independent of both sperm and egg genotype. No significant differences were detected between groups of eggs from 5 strains of mice fertilized by spermatozoa from a single strain, TO, or between groups of (C57BL/10 X CBA) F1 eggs fertilized by spermatozoa from 2 strains, CBA/H-T6 and TO, although there was significant heterogeneity between individual males in the latter set of experiments. While the majority of polyploids were triploids resulting from dispermic fertilization, some derived from diploid spermatozoa were also found.


Effect of tolerance to paternal antigens on placental and fetal weight in the mouse. Placental and fetal weight was measured on the 16th day of pregnancy in normal and tolerant mice. Placental weight was increased in A mice tolerant to C57BL/10 mice only when donor cells from males were used. Fetal weight was unaffected.


Effect of implantation delay on transfer of rat embryos to mice. Rat and mouse blastocysts were transferred to the uteri of ovariectomized mice, maintained by progesterone for a few days and induced to implant by oestradiol administration. Mouse implantation sites contained normal embryos; all rat embryos were retarded, although some had developed to the egg-cylinder stage.


Presence of oestradiol-17beta in the rabbit ovum and its investments. Untreated rabbit eggs contained 1-2 pg oestradiol-17beta/egg. Treatment with hyaluronidase for 20 min, but not for 10-13 min, removed the cumulus and coronra cells and all oestradiol.


The parental source of heteroploidy in chick embryos determined with chromosomally marked gametes. Males homozygous for chromosomal translocations were mated to karyologically normal females. The resulting embryos, after incubation for 16-18 h, were prepared for cytogenetic analysis. Of the 62 embryos analysed, 54 (7.8%) were heteroploid or contained a major heteroploid cell line. Each of the 638 normal 2n embryos contained one marker and one normal chromosome, indicating that none arose from gynogenesis. Thirteen homogeneous haploid embryos were identified; 24 embryos were chimaeric 1n/2n and one was 1n/3n. All haploid cell lines contained a marker chromosome indicating androgenetic origins. The 9 homogeneous triploid (3n) embryos and the 3n cell line in a single 1n/3n embryo contained a single marker. All resulted from fertilization by single spermatozoon of eggs that were diploid as a result of suppression of the second meiotic division. The 3n lines of two 2n/3n embryos were derived from other mechanisms. A single homogeneous tetraploid (4n) embryo and the 4n cell lines of three 2n/4n mosaic embryos each contained two marker chromosomes and presumable resulted from failure of cytokinesis in an early cleavage division.


Prostaglandin concentrations in the semen of fertile men. The PG concentrations in the semen of 23 fertile men were 73 microgram PGE/ml, 267 microgram 19-OH PGE/ml, 2.1 microgram PGF/ml and 18.3 microgram 19-OH PGF/ml. The wide ranges of concentrations found for the PGEs (2-272 microgram/ml) and for the 19-OH PGEs (53-1094 microgram/ml) throw some doubt on the previously established correlation between infertility and low prostaglandin concentrations.


The effect of rete testis fluid on the metabolism of testicular spermatozoa. The O2 uptake of spermatozoa collected from the rete testis of conscious rams and boars has been measured polarographically either in the rete testis fluid (RTF) in which they were collected or in a synthetic saline medium (S-RTF) made to resemble the ionic composition of RTF as closely as possible. In both species, O2 uptake of the testicular spermatozoa was higher in RTF than in S-RTF. The addition of some of the organic constituents of RTF was without effect on O2 uptake. Pyruvate, lactate, acetate or alanine increased the O2 consumption of ram spermatozoa in S-RTF. Lactate had no effect on but acetate increased the O2 consumption of pig testicular spermatozoa. Phospholipid phosphorus and fatty acids decreased during incubation of ram spermatozoa in RTF and S-RTF. The addition of substances which increased the O2 consumption of the spermatozoa in S-RTF was without effect on the lipid composition. The fatty acid composition of the phospholipids and the proportions of the various phospholipids were unaffected by incubation. These results indicate that rete testis fluid has an appreciable effect on the metabolism of the spermatozoa it contains, and that this effect cannot be mimicked by any of the known organic constituents of the fluid.


In-vitro production of prostaglandins by intrauterine tissues from pregnant goats. In the fetal cotyledons and membranes, PGE production (determined in a continuous superfusion system) was significantly greater than that of PGF (P less than 0.01 and P less than 0.02 respectively). The fetal cotyledon also produced more PGE than the maternal cotyledon (P less than 0.01) and more PGF after parturition than before (P less than 0.01).


A reversible contraceptive action of some 6-chloro-6-deoxy sugars in the male rat. Male rats given daily doses of 240 mumol 6,6'-dichloro-6,6'-dideoxysucrose or 6'-chloro-6'-deoxysucrose or 6-chloro-6-deoxysucrose/kg for 28 days became infertile after 7 days and remained infertile during treatment. Fertility was partly recovered 3 weeks after the last dose and completely restored by 6 weeks. 6-Chloro-6-deoxyglucose and 6-chloro-6-deoxyfructose were also effective at this dose, but 6,1',6'-trichloro-6,1',6'-trideoxysucrose, 4,1',6'-trichloro-4,1',6'-trideoxygalactosucrose,4,6,-4',6'-tetrachloro-4,6,4',6'-tetradeoxygalactotrehalose and methyl-6-chloro-6-deoxy-alpha-D-glucopyranoside were not. Spermatozoa from the cauda epididymidis of infertile rats oxidized glucose more slowly than did spermatozoa from controls.


Induction of avidin in the chick oviduct by tissue damage and prostaglandins. In immature, diethylstilboestrol-treated chicks, ligation of the oviduct caused local avidin synthesis in the immediate vicinity of the ligature. PGF-2alpha injected directly into the oviduct also induced avidin synthesis, whereas saline or PGE-2 had no effect. PGE and PGF-2alpha concentrations increased in the oviduct within 24 h of ligation: the PGE increase could be partly inhibited by indomethacin, whereas that of PGF-2alpha was less inhibited. An LD50 dose of indomethacin alone and with ligation had a clear stimulatory effect on avidin synthesis, whereas aspirin alone, or with ligation, was not effective. Ligation alone and with indomethacin appeared to alter the PGF-2alpha/PGE ratio. These results suggest that PGF-2alpha may be involved in the regulation of avidin synthesis in the chick oviduct.


Movement characteristics and power output of guinea-pig and hamster spermatozoa in relation to activation. Spermatozoa were collected from the cauda epididymidis of golden hamsters and guinea-pigs, and the acrosome reaction was induced in vitro. Movement characteristics of the spermatozoa were assessed with high-speed cinemicrography. Before the initiation of the acrosome reaction (preactivated spermatozoa), sperm movement in both species was characterized by progressive swimming by regular flagellar waves of moderate amplitude and relative high frequency. After the acrosome reaction (activated spermatozoa), sperm movement in both species was not progressive, and was characterized by whiplash-like flagellar undulations of significantly (P less than 0.05) higher amplitude and lower frequency. Calculation of the hydrodynamic power output by a new theory indicated that no significant change occurred after activation.


Induction of testicular growth and sexual activity in rams by a 'skeleton' short-day photoperiod. Eight adult rams were housed in 16L:8D for 16 weeks and then exposed to short days (8L:16D) or 'skeleton' short days (11L:1D:5L:7D) for 16 weeks before being returned to long days. The 'skeleton' treatment promoted testicular development and regression in a way similar to that occurring in 8L:16D, indicating that a change in the total quantity of light is not a prerequisite for the photoperiodic response in the ram.


Maturation of the pituitary-gonadal system in the male rat. Serum FSH and testosterone concentrations reached maximum levels between 35 and 45 days of age, which coincided with the appearance of mature spermatozoa in the majority of seminiferous tubules. Spermatozoa were not observed in sections of the urethra until the age of 46 days. Serum LH concentrations were low (5-6 ng/ml) before Day 25, became highly variable (12-57 ng/ml) between Days 25 and 53 and remained consistently above 35 ng/ml thereafter. Serum prolactin levels rose significantly between 30 and 43 days of age. Maximum prolactin levels coincided with the start of accelerated growth in the prostate and seminal vesicle glands. Testicular weights relative to body weight reached a plateau by 35 days of age, while relative pituitary and adrenal weights decreased throughout the study period. It is suggested that spermatogenesis is not complete until FSH and testosterone reach maximum levels, while prolactin may be involved in the stimulation of accessory sex organ growth. The pronounced variation in serum LH concentrations during the maturation period may reflect a progressive change in the sensitivity of the hypothalamic-pituitary axis to the negative feedback of gonadal steroids.


Radioimmunoassay of FSH in the plasma of post-partum dairy cows. A method for measurement of bovine plasma FSH has been established with an interassay coefficient of variation of 15.5% over the workable range of the assay. Compared to values at the end of pregnancy (42-122 ng/ml), FSH concentrations were greater (P less than 0.01) between 0 and 20 days post partum in 3 cows.


Plasma progesterone levels during delayed implantation in the European badger (Meles meles). There was a biphasic pattern of progesterone secretion during the year. Delayed implantation was characterized by low concentrations from February to June, a significant (P less than 0.001) increase during July, August and September, and a return to low levels in October-November. A second significant increase (P less than 0.001) was observed in December and early January just before the presumed time of implantation.


Androgens and androgen-binding protein in the rat epididymis. The levels of testosterone, dihydrotestosterone (DHT), 5alpha-androstan-3alpha,17beta-diol and androgen-binding protein (ABP) were measured in various segments of the epididymis from adult rats which had been unilaterally orchidectomized for 4 weeks. On the 'intact' side, ABP concentrations were highest in the caput region. The segmental distribution of DHT closely followed that of ABP with the highest concentration in the caput (40 ng/g tissue) and lowest in the cauda (10 ng/g tissue) epididymidis. There was a high degree of correlation (r = 0.98) between the concentration of DHT in the epididymis and ABP levels. 'Castration' completely abolished the DHT gradient. The levels of testosterone and androstanediol were lower than those of DHT; most was present in the corpus epididymidis. The relative differences were reduced after 'castration'. It is concluded that ABP in the rat epididymis is the primary factor for determining the concentration of DHT in the epididymal fluid.