Toxic metabolites of Amanita pantherina, A. cothurnata, A. muscaria and other Amanita species. Forty-seven specimens representing 35 species or varieties of Amanita were examined for the presence of ibotenic acid, muscimol, stizolobic acid, stizolobinic acid, aminohexadienoic acid and chlorocrotylglycine. In addition, crude extracts of A. muscaria were examined chromatographically for the presence of methyltetrahydrocarboline carboxylic acid (MTC). Ibotenic acid and muscimol were found in clearly detectable concentrations in extracts of A. cothurnata, all specimens of A. muscaria, all specimens of A. pantherina and in lower concentrations in A. gemmata. Stizolobic acid and stizobinic acid were found in detectable concentrations in one variety of A. muscaria, in all specimens of A. pantherina and in trace levels after additional purification of the extracts in A. gemmata and in the remaining specimens of A. muscaria. Aminohexadienoic acid and chlorocrotylglycine were detected only in crude extracts of A. smithiana. MTC could not be detected in crude extracts of A. muscaria. Crystalline ibotenic acid (820 mg) was isolated from 17 kg of a specimen of A. pantherina collected in western Washington State.


Biological and phytochemical evaluation of plants. XIV. Antiinflammatory evaluation of 163 species of plants. One hundred and seventy-seven plant extracts, representing 163 species of plants and/or fungi, were evaluated in rats to determine their antiinflammatory activity using the carrageenin-induced pedal edema assay. Of the 163 species of plants and/or fungi tested, 17 exhibited between 30--39 per cent inhibition of inflammation, 21 between 40-49 per cent, 15 between 50--59 per cent, four between 60--69 per cent, and two gave greater than 70 per cent inhibition of carrageenin-induced pedal edema.


Orphan data and the unclosed loop: a dilemma in PSRO and medical audit. The experience of the Utah Professional Review Organization in conducting medical audit as a PSRO prototype, and later a PSRO, suggests that usage of diagnosis-oriented process audits, using criteria similar to those supplied by the American Medical Association, will result in accumulation of vast amounts of unusable data. Even studies based on outcome data may fail to change physician behavior substantially because of communication difficulties between the PSRO or hospital conducting the evaluation and the physicians subject to the audit. UPRO's experience suggests that medical audit can be made more effective if it employs a special study format that looks at a particular element of care, selected for its importance to patient welfare and potential for improvement. The study should be based on criteria solidly validated by clinical research, and should involve a direct and personal interaction between the review body and the physician being reviewed.


Supersensitivity of cortical neurones of the rat to acetylcholine and L-glutamate following chronic morphine treatment. the effect of microelectrophoretically applied L-glutamate and acetylcholine on discharge activity of cortical neurones was studied in naive and in morphine-tolerant/dependent rats. The thresholds for increase in discharge activity elicited by these 2 putative neurotransmitters were 3 times lower in the tolerant/dependent rats than in the naive rats, indicating the development of supersensitivity.


Circular dichroism studies on the interaction of sulfonylureas with insulin. The interaction of 5 sulfonylurea derivatives with insuline was investigated by means of circular dichroism measurements. It was found that all sulfonylureas investigated decrease the ellipticity band of insulin at 208 nm, whereas the band at 222 nm remains unaffected. It is suggested that these observations are due to a change of the insulin conformation, provoked by the interaction of the drugs with insulin. It is assumed that such an effect on the insulin conformation can influence the binding properties of insulin, e.g. in respect to the insulin aggregation, to the binding to insulin antibodies and to a bound, inactive form of insulin in the plasma. Some other drugs have similar, but mostly smaller effects on the CD spectrum of insulin. Therefore, the importance of these observations for the hypoglycaemic action of the sulfonylureas is not yet clear.


[Disturbance in fluid and electrolytes metabolism with central origin with special reference to sodium (author's transl)]. Clinical aspects with disturbances in fluid and electrolytes metabolism in brain diseases were discussed reviewing 41 cases experienced in our department. These 41 cases were found in 377 patients with diseases of the central nervous system in our hospital during recent 14 months. Hyponatremia was found in 19 cases and aneurysms of A-C, A1 and A2 had the majority of the cases. The cerebral angiography suggested an unstable blood supply to the anterior portion of the hypothalamus, for instance, showing remarkable shift, spasm or obstruction A-C, A1 or A2. The duration of hyponatremia was transient and mostly less than 2 weeks after the last attack of subarachnoid hemorrhage. On the contrary, hypernatremia was seen in 9 cases and 6 of them were found in cases of tumors in the pineal region and A-C, A1 and A2 were intact angiographically. The hypernatremia was continuous and did not response to V-P shunt or any kinds of infusion therapy. The hypernatremia due to cerebral disease is thought to be a result of destruction of the supraoptic and paraventricular nuclei or adjacent area in the anterior potion of the hypothalamus in most of presumed these cases. It might be that the decreased blood supply to the anterior position of the hypothalamus offers an information not of hypoosmolarity but of hypovolemic state, and this information increases the secretion of ADH. This mechanism of hyponatremia could play an important role in S.I.A.D.H.


Endocrinological evaluation of sellar and suprasellar tumor cases (the eighth report)--on growth hormone secretion of acromegalic patients before treatment (author's transl). Serum GH levels were measured in 14 acromegalic patients in ITT, TRH test, LH-RH test, arginine test, OGTT, exercise test, CPZ test and PEG. More than 50% increase of serum GH level was observed in 3 of 13 ITT, 8 of 14 TRH test, 6 of 14 LH-RH test, 2 of 12 arginine test, 2 of 9 OGTT and 2 of 7 PEG. Only 2 out of 7 cases showed more than 50% decrease of serum GH level in CPZ test. Striking GH responses were observed in TRH and LH-RH test. Five of 14 cases showed more than 100% increase of serum GH level in TRH test but they did not have a significant response to LH-RH. Four of 14 cases also had more than 100% increase in LH-RH test with no significant response to TRH. The rest of the cases (5 cases) did not have marked response to either TRH or LH-RH. According to these results, serum GH responses of acromegalic patients could be devided into 30 types, 1) TRH responding type, 2) LH-RH responding type, 3) no response type.


[Method of superimposing the angiographically located supratentorial lesion on the scalp prior to craniotomy]. One of the important points in operation of the intracranial supratentorial lesion is appropriate site and size of bony window made by craniotomy and this matter is also important first step in procedure of craniotomy. On the other hand, the site and size of bony window made in craniotomy for supratentorial lesion has relationship with perfectibility of operation. The detail intracranial situation and extent of supratentorial lesion is decided from the datas of various examinations and the findings in cerebral angiogram give a most important and valuable information to the neurosurgeon at present. The neurosurgeons used to decide the area of craniotomy from the findings of cerebral angiogram but there are some difficulty in transfer of findings related with situation of supratentorial lesion in angiogram to the patient's scalp, because the film of cerebral angiography is a projected picture of spheric head by X-ray to the plane. The author devised the planning method of site and size of bony window in craniotomy by transfer the location and extent of supratentorial lesion in cerebral angiogram to the patient's scalp and the author have been recognized for the past five years that this method is simple one and has clinical accuracy. The principle of the author's method are as follows. The film of cerebral angiography in lateral projection and the patient's scalp are divided into nine parts by same manner and the relation of the site and extent of lesion in cerebral angiogram with divided parts transfer to the division of the patient's scalp under special care to make minimize errors due to use the cerebral angiogram which is picture made by projection in a plane from spheric intracranial supratentorial space. Five points and seven lines are used to divide the film of cerebral angiography and the patient's scalp. Five divide points are most upper part of margin of external acoustic meatus, most posterior edge of auricle, upper, lower and lateral edge of orbit. The following divide lines are drawn by use of these five divide points. Two horizontal lines are eye-ear horizontal line and superior eye horizontal line. Three vertical line are external acoustic meatus vertical line, auricular vertical line and temporal orbital margine vertical line. Two oblique lines are anterior and posterior oblique lines. The film of lateral cerebral angiography and patient's scalp are divided into nine parts by these seven divide lines. In the areas above the superior eye horizontal line, following procedure for correction of error due to transfer the point of X-ray film to the patient's scalp is needed because of strong curved brain surface. One end of celluloid or steel scale place closely with the scalp below the superior eye horizontal line and the other end of scale make freely from curved scalp surface. The point of lesion on the divide line of X-ray film marks at correspond point on the scale...


[Observation of the epicerebral microcirculation studied by fluorescein angiography (author's transl)]. Fluorescein angiography was carried out by modification of method as described by Feindel et al. The hemisphere was widely exposed to allow visualization of surface blood vessels. In the experimental room, the dye was injected rapidly either through a fine polyethylen catheter placed in the lingual artery or thorough the cannula in the femoral vein. For "lingual" angiography 1.6 ml of 1% sodium fluorescein were used, while for "femoral" angiography, 4 ml of 10% solution were injected. In the operating room 4 ml of 1% sodium fluorescein are rapidly injected into an internal carotid catheter. Serial photographs of the passage of the dye through the surface vessels of the hemisphere were taken with a motor-drive Nikon camera at interval of 0.4 seconds, or longer when indicated, starting at the time of the injection. The shutter was synchronized with the discharge of a rapid re-charging stroboscopic light. A wratten gelatin filter 47A (Kodak) was used over the light and a Nikon filter Y52 over the camera lens for Kodak high speed ektachrome film to obtain color photographs. The timing of the interval between photographs was measured to within in 0.4 seconds by recording from a ink written oscillography synchronized with the shutter. Thus the velocity of flow in individual vessels could be calculated from the serial photographs.


[Spontaneous spinal epidural hematoma (author's transl)]. A case of spontaneous spinal epidural hematoma during pregnancy is reported. Although we operated the patient 14 hours after the initial symptoms, the functional prognosis was poor. It is believed that the spinal cord was damaged irreversibility because of the rapid development of spinal cord edema. Referring to the reported cases we discussed symptoms, diagnosis and therapy as well as etiology. As to our case it is supposed that the rise of epidural venous pressure due to the pregnancy caused the rupture of the epidural venous plexus. About spontaneous spinal epidural hematoma during pregnancy, so long as we examined, there is only one case report, which Bidzinski reported in 1966.


[Homonymous hemianopsia due to chiasmal compression by intracranial aneurysm (author's transl)]. Eight cases of homonymous hemianopsia due to chiasmal compression by intracranial aneurysms have been reported in the literatures. Two similar cases were experienced in our clinic and reported. Of the total 10 cases, 8 were giant aneurysms--over 2 cm in diameter--and 9 cases were aneurysm developing medially from the internal carotid artery. Visual disturbances in all the 10 cases were incongruous homonymous hemianopsia without macula sparing. It should be noted that the existence of aneurysm is possible, in patients with homonymous hemianopsia, even when there is no other clinical evidence of mass lesion.


[Nasopharyngeal extension of a large chromophobe adenoma of the pituitary (author's transl)]. It is not common to have experienced the nasopharyngeal extension of pituitary adenomas. Recently we have experienced such a case. A man, aged 18, height 168 cm, weight 66 kg, who admitted to the hospital with the complaints of headache, left nasal obstruction, loss of visual acuity and defect of his temporal fields. On examination of both fundi there was primary optic atrophy. At this time large tumor could be seen in the nasopharyngeal cavity. Plain X-ray showed that the pituitary fossa was definitely enlarged and that there was considerable destruction of the sella and the clivus. Definite soft tissue mass could be visualised clearly by tomography. Via transoral and transsphenoidal approach, total removal of the nasopharyngeal tumor and intracapsulary subtotal resection of the tumor were performed. Microscopical examination established the diagnosis of chromophobe adenoma. On postoperative examination of hypothalamopituitary function he had no responces to insulin hypoglycemia and arginine infusion in growth hormone. 60Co irradiation, totally 6,000 rad, was given. Two years postoperatively he showed fairly good deal of improvement of his visual field defect. There was no serious complaints other than visual impairment on the left eye.


[Successfully operated case of posterior fossa tuberculoma in childhood (author's transl)]. A case of intracerebellar tuberculoma is described in which a tuberculoma was removed successfully through the administration of antituberculous agents, and a full recovery was obtained. The patient was a 3-year-old boy who had been receiving antituberculous agents for about 4 months because of acute inflammation followed by osteomyelitis of his right big toe, which was suspected to be tuberculous, and because of pulmonary tuberculosis diagnosed in a chest roentgenogram taken about 1 month after osteomyelitis was cured. While his osteomyelitis was being treated, disturbance in his gait, due to progressive spastic paraparesis, was not iced, and thereafter left cerebellar symptoms with impairment of equilibrium appeared additionally. Then, he was reffered to our clinic for further neurosurgical examination, and was admitted on November 1, 1974 after right carotid and vertebral angiography was performed via the right axillar artery, in which findings suggesting left cerebellar tumor and internal hydrocephalus were obtained. After he was admitted to our clinic, a diagnosis of tumor of the left cerebellum and internal hydrocephalus was more precisely confirmed by pneumoventriculography. Suboccipital craniectomy was then carried out and the tumor, weighing 60 gm, was completely removed from the left cerbellar hemisphere. The tumor was confirmed as tuberculoma not only by histological findings but also by the vertification of tuberculous bacilli in it. Though, moderate fever lasted for about 2 weeks postoperatively, no obvious meningitic signs or new neurological deficits were noted. The patient showed marked improvement especially in his gait disturbance, and was discharged ambulatory 40 days after the operation, and has since been asymptomatic except for slight ataxic gait. The antituberculous agents have been continuously administered postoperatively.


Optimal utilization of computerized cranial tomography and radionuclide brain imaging. The results of computerized cranial tomography and radionuclide brain imaging in 490 patients were compared in relationship to the patients' clinical presentation. In 195 patients with focal neurologic abnormalities, both tests detected most lesions, but computerized cranial tomography was slightly more accurate overall. Results of both studies were normal in 69 percent of 295 patients with nonfocal neurologic presentations, and radionuclide imaging failed to detect lesions in only five patients with nonfocal presentations. These results suggest that radionuclide imaging can be used to accurately screen most patients with nonfocal neurologic presentations. An exception is the patient presenting with dementia, in whom computerized cranial tomography provides details of the anatomy of the ventricular cavities and cerebral cortex. This study demonstrates a continuing role of importance for radionuclide imaging in the evaluation of patients with neurologic disease and provides data to allow a rational approach to the optimum use of both techniques.


Postural hypoxemia in quadriplegic patients. Quadriplegic patients frequently undergo unusual positional changes, including head-down tilt. To determine if hypoxemia develops in this position and whether it is related to the duration of quadriplegia, five patients were studied within 1 year of injury (group A) and five after 1 year from injury (group B). Arterial blood gases were obtained in the seated and 45-degree head-down position. Group A patients had a decrease in arterial partial pressure of oxygen in the head-down position (p less than 0.025), while group B patients did not (p greater than 0.4). This study demonstrated that quadriplegic patients have positional hypoxemia early in their course. This probably results from relative hypoventilation of the upper lung fields caused by flaccid paralysis of the intercostal muscles. The positional hypoxemia disappears with time, presumably because of the change from flaccid to spastic paralysis of the intercostal muscles.


Sex difference of multiple sclerosis in Japan. Sex differences in the clinical course of multiple sclerosis were investigated in 947 patients with multiple sclerosis collected from all over Japan through the nationwide survey undertaken by the Multiple Sclerosis Research Committee of Japan from 1972 through 1973. A course of remissions and exacerbations, more frequent involvement of optic nerves, and more severe motor and visual disability were present in a higher proportion of women than men. These features corresponded to those which have been considered to characterize Oriental multiple sclerosis patients.


Normocalcemic tetany. A problem of erethism. A boy with neonatal and childhood convulsions had prolonged attacks of tetany in adolescence. There was no abnormality of serum calcium or magnesium, and treatment with these cations was ineffective. There was no respiratory alkalosis, and attacks occurred when the patient had not taken anticonvulsant drugs for years. Serum parathormone content and renal responses to the administration of parathormone were normal. "Normocalcemic tetany" seems an appropriate name for the condition, which was probably genetic since the patient's brother and mother had signs of latent tetany and the brother had a convulsive disorder. The cause of the syndrome is not known, but it seems to be an abnormal response of neural membranes rather than an abnormality of calcium homeostasis.


Cerebrovascular carbon dioxide reactivity and conductance in patients awake and under general anesthesia. Paired cerebral blood flow measurements at two levels of arterial carbon dioxide (CO2) tension were made in 36 awake patients and 85 patients under general anesthesia to measure the percent change in flow per mm Hg change in PaCO2 (CO2 reactivity). CO2 reactivity was significantly greater in the generally anesthetized (5.96) than in the awake group (3.65). In both groups, a linear correlation was found between mean percent CO2 reactivity and conductance (the reciprocal of resistance), up to a level of conductance of 0.85 in awake and 0.95 in unconscious patients. When the CO2 reactivity of 18 awake patients and 59 unconscious patients was compared with that of respective control groups, only 11 patients were found to have abnormal CO2 reactivities for their conductance levels.


Recovery in rats after spinal cord injury. Previous studies from this laboratory have shown evidence of regeneration of long descending spinal motor tracts in rats after spinal cord transection and treatment to modify the animals' immune response. In this study, less extensive surgical lesions were combined with the most favorable drug treatment (75 mg per kilogram of cyclophosphamide in a single dose) in an effort to improve the prospects for regeneration. Less than complete spinal cord transections in the rat were frequently followed by clinical and electrophysiologic evidence of return of function. Such return of function appears to depend on a reorganization of the nervous system that results in the use of the few remaining fibers to transmit motor information rather than on regeneration. Immunosuppressive treatment had no effect on these results.


Quantitative histochemical evaluation of normal human skeletal muscle. Skeletal muscle tissue was obtained by open biopsy from the vastus lateralis and peroneus brevis muscles from 12 and 16 healthy paid volunteers, respectively. Frozen sections were examined with standard histochemical methods. Central nuclei, small and large angular fibers, and small round fibers were the most common "abnormalities" present. The number of fibers of these types were quantified, along with other more rare deviations from normal morphology. Several of the abnormalities were more common in the peroneus brevis than in the vastus lateralis.


Incidence of Z band streaming and myofibrillar disruptions in skeletal muscle from healthy young people. Skeletal muscle tissue was obtained from the vastus lateralis or peroneus brevis muscles of 34 healthy volunteers. Focal and extensive areas of Z band streaming and disruption of myofibrillar architecture without Z band streaming were quantified in Araldite-embedded specimens examined in the phase microscope. Extensive areas of Z band streaming in more than 2.0 percent of fibers was present in only two of the 34 volunteers. Myofibrillar disruptions without Z band streaming was much less common than Z band streaming. The size of extensive areas of Z band streaming or myofibrillar disruptions was quantified by counting the number of adjacent fibrils and contiguous sarcomeres occupied by either type of lesion. These data should be of use in determining if such lesions are present in excessive amounts in suspected pathologic material.


Congenital retardation and central motor defect with later evolution of seizure disorder, orofacial dysplasia, and amyotrophy. A clinicopathologic report. Retarded mental and motor development was observed during the first year of life of a full-term female who had been delivered with difficulty. Generalized psychomotor seizures and dysplastic "acromegalic" facial changes began to develop when she was 8 years old, and generalized amyotrophy developed over the next several years. The course was generally progressive, and she died at age 31. A variety of clinical laboratory studies were nondiagnostic. Autopsy findings showed normal brain weight and normal-appearing cerebral hemispheres, a peculiar atrophy of the inferior portion of the cerebellar hemispheres, a demyelinating process in the dorsal columns of the cervical and thoracic spinal cord, and lateral column and motor neuron degeneration. The pathogenesis of the syndrome is undetermined.


Use of phenytoin serum levels in a case of status epilepticus. Serum anticonvulsant determinations made possible the identification of an unusual cause of status epilepticus and assisted with appropriate therapy in an epileptic patient with second-degree burns. The seizures were associated with a serum phenytoin concentration of 3.5 mug per milliliter. This suboptimal concentration of drug developed despite continuation of the patient's customary oral dose of phenytoin led to the conclusion that the unexpected decrement in serum anticonvulsant concentration was related to an experiment in absorption of that drug, possibly caused by the concurrent administration of oral oxacillin.


Platelet aggregation, stroke, and transient ischemic attack in middle-aged and elderly patients. Platelet aggregation was studied with optical density methods in a group of 39 patients with stroke or transient ischemic attacks (TIA) and in age, sex, and race-matched controls. The patients were divided at age 60 into young stroke patients and young controls (18 pairs) and old stroke patients and old controls (21 pairs). A semiquantitative measure of the threshold of phase II of platelet aggregation and distinct of disaggregation 3 minutes after peak aggregation were used as an index of platelet aggregability. Aggregability was significantly greater in young stroke patients than in young controls. Aggregability was similar in old stroke patients and old controls. Both old stroke patients and old controls were hyperaggregable compared with young controls, indicating that aggregability rises with age. This suggests that platelet aggregability is significant risk factor for stroke but is relatively more important in the younger than in the older stroke patient.


Phosphorylase deficiency associated with isometric exercise intolerance. In the patient reported here, most of the clinical and biochemical correlates of McArdle's disease were present; however, symptoms developed after isometric rather than after isotonic exercise. Histochemical reactions and assay of muscle homogenates confirmed the absence of phosphorylase; however, venous serum lactate increased in the arm after isometric and isotonic ischemic exercise, although the increase was significantly less than in a normal control. Although the symptoms differ significantly from those in McArdle's disease, this disorder could be a variant.


Basophilic meningitis secondary to lymphoma. In this case report of basophilic meningitis, the patient was a 5-year-old girl under treatment for a diffuse lymphocytic lymphoma. She presented with headache and bilateral papilledema. Cerebrospinal fluid examination showed 60 percent basophils. Subsequent specimens showed a rare blast. It is postulated that after the lymphoma cells had spread to the meninges, a cell-mediated immune reaction was initiated with the appearance of basophils in the exudate.


[Primary lung neoplasm of rare incidence: giant cell carcinoma]. Primary giant-cell carcinoma of the lung is an infrequent observation. It is distinguished clinically solely its fairly rapid development but otherwise it does not seem to have any predilection for localization, age or sex. Histologically, differential diagnosis with othe tumours such as angiosarcoma, rhabdomyosarcoma, chorioepithelioma and others is of interest. Three cases of primary giant-cell carcinoma of the lung are reported and the various clinical and anatomopathological aspects discussed.


Severe lactic acidosis associated with intravenous alcohol for premature labor. A patient receiving intravenous alcohol for premature labor developed mild aspiration pneumonia. In the course of her evaluation it was noted that she also had severe lactic acidosis. The patient responded well to hydration and intravenous sodium bicarbonate. Lactic acidosis represents a previously unrecognized danger of intravenous alcohol therapy.


Applying management principles to the occupational health unit. This paper presents the occupational health unit as a subsystem competing with other subsystems in the organization for limited resources. By applying management principles the nurse can design a program to show management its results in dollar terms. This will enable the health unit to compete more favorably with the other subsystems for limited resources. An example of how this approach can be applied to the problem of absenteeism is presented.


Microbiological quality of frozen fried chicekn product obtained from a retail store. Two brands of frozen fried chicken products were purchased monthly from a local supermarket for six months. Microbiological qualities of these samples were studied. The log number of mesophilic counts ranged from 2.90/g. to 4.78/g; log psychrophilic counts varied from 2.74/g. to 4.66/g.; and log Staphylococcus- 110 medium counts ranged from 2.84/g. to 4.54/g. Mean log microbial counts were higher in brand A samples than in brand B samples. No yeast or mold was detected and all samples were Salmonella negative. Most samples were negative in coliform test, except five samples had coliform MPN ranging from 0.5/g to 0.8/g. A total of 144 isolates of psychrophiles from 12 samples was tentatively identified to be members of Staphylococcus species. About 82.2% of the isolates belong to the Subgroup VI Staphylococcus according to Baird-Parker's classification. Another 17.8% of the isolates resembled Subgroup vi Staphylococcus except in phosphatase test.


Schedule-controlled behavior in the morphine-dependent rat. The behavioral effects of acute and chronic administration of morphine and its withdrawal were studied using schedule-controlled responsing in the rat under a differential reinforcement of low rates of responding (DRL) schedule of food presentation. Acute morphine administration had a biphasic effect on response rate. Low dses (1.8-5.6 mg/kg) tended to produce a small increase and higher doses (10-30 mg/kg) decreased responding. Physical dependence was produced by twice daily injections, with an initial dose of 40 mg/kg/day which was increased by 80 mg/kg/day until reaching 600 mg/kg/day which was continued for 14 days. Throughout chronic administration the pattern of responding remained disrupted resulting in a 27-47 percent decrease in presentations of the reinforcer, while response rate was more variable and generally decreased. The effects of morphine withdrawal lasted 5 days and produced an initial maked decrease in reinforecements per hour and a biphasic change in response rate. A marked decrease in responding early in withdrawal (22.5 hr) was followed by a marked and more prolonged (70.5-118.5 hr) response rate increase.


Behavioral and biochemical effects of neonatal treatment of rats with 6-hydroxydopa. Rats receiving injection of either 6-hydroxydopa (60 mug/g) or saline on Days 1, 3, and 5 of life were studied in adulthood on a number of behavioral tasks before being sacrificed at 8 or 12 months for NE assay. The treated rats exhibited impaired passive avoidance, less shock-induced aggression, and more locomotor open-field activity than the control rats. There were no differences between the groups in male copulatory behavior, food and water intake, or thermoregulation. In comparison to the saline rats, 6-hydroxydopa rats showed elevated levels of endogenous NE in lower brainstem regions, e.g., midbrain and pons-medulla, as well as cerebellum. Hypothalamic NE level was not affected. Significant depletions of NE were obtained in the hippocampus and neocortex.


Sites of action of morphine involved in the development of physical dependence in rats. I. Comparison of precipitated morphine withdrawal after intraperitoneal and intraventricular injection of morphine antagonists. In rats made dependent on morphine by implantation of morphine pellets, withdrawal, as precipitated by intraventricular injection of morphine antagonists, was compared to withdrawal as precipitated by systemic antagonist application. The results, most clearly those obtained with a hydrophilic compound, diallyl-nor-morphinium-bromide, point to periventricularly located sites of action for the release of most withdrawal signs by antagonists. Jumping, reaching only low levels after i.ventr. injection of levallorphan and nalorphine, was very pronounced when the benzomorphane derivative SH 254, was used. In the case of writhing and diarrhea, the situation is more complicated. Possibly, central as well as peripheral mechanisms are involved in the expression of these signs.


Sites of action of morphine involved in the development of physical dependence in rats. II. Morphine withdrawal precipitated by application of morphine antagonists into restricted parts of the ventricular system and by microinjection into various brain areas. Morphine withdrawal was precipitated by injection of various morphine antagonists into restricted parts of the ventricular system or by microinjection of levallorphan into specific brain areas of rats made dependent on morphine by repeated pellet implantation. When the antagonists could spread only within the lateral ventricles and the 3rd ventricle, a weak withdrawal syndrome was induced; by antagonist administration into the restricted 4th ventricle, however, strong withdrawal signs like jumping were elicited even at small dosages. In microinjection experiments, structures in the midbrain and the lower brain stem proved to be the most sensitive to antagonist action. Although microinjections into thalamic nuclei also had some effect, it could not be excluded that the effects were due to uncontrolled spreading of the drug. This became especially clear from experiments with tritium-labeled levallorphan. It is concluded that brain structures located in the anterior parts of the floor of the 4th ventricle and/or caudal parts of the periaqueductal gray matter are important sites of action for the development of physical dependence on morphine.


AMT catalepsy and hypokinesia: interaction with morphine and cocaine. Acute morphine induced a dose-dependent hypokinesia and rigidity, but only mild and non-dose-dependent catalepsy. AMT, injected 1/2 h after morphine, slightly potentiated catalepsy but not hypokinesia during 3 h after morphine; in contrast, rigidity was decreased. The behavioral changes induced by AMT were accelerated in onset and reached their usual development, although AMT toxicity and hypothermia were completely antagonized; thus, it would appear that AMT hypokinesia/catalepsy are not the consequence of toxicity. When morphine was injected 4 h after AMT, a mutual potentiation of the two drugs on hypokinesia and catalepsy was observed, although previous biochemical measurements had shown no effect of morphine on CA depletion under these conditions. Rigidity appeared to be antagonized. After 17 days of repeated injections, morphine no longer elicited hypokinesia and catalepsy, but no cross-tolerance developed to the AMT behavioral changes. A similar lack of cross-tolerance to the effects of AMT or haloperidol was observed when morphine tolerance was induced by pellet implantation. Catalepsy and hypokinesia developed in a much more pronounced way after two large i.p. doses than after small, multiple administration of AMT; this difference was accompanied by a significantly lower concentration of brain DA, but not NA in the former group. The hyperthermic response observed after a 40 mg/kg s.c. injection of morphine was reversed to hypothermia when the same dose was given 4 or 10 h after CA synthesis inhibition. Cocaine strongly antagonized AMT hypokinesia and catalepsy when given 8 1/2 h after AMT, and, although to a lesser extent, even when injected 12 1/2 h after AMT.


[Amantadine: aggressive effect; effect on apomorphine-induced aggressivity in the rat]. In rats characterized by aggressive behaviour induced by apomorphine, only amantadine in doses of 10 mg/kg i.v. and 50 mg/kg i.p. induces a very slight aggressive effect, but in doses of 30 and 50 mg/kg i.p. it inhibits the aggressive behaviour induced by 1 mg/kg apomorphine. The effect is dose-related and it is more pronounced when amantadine (50 mg/kg i.p.) is injected 30 min rather than 1 h before apomorphine.


Increased sensitivity to chronic ethanol in isolated mice. Isolated C57BL/10 mice fed liquid diet as their only nutritional supply consumed 44% more diet than did grouped-housed mice. A similar increase due to isolation of 36% for C57BL/10 mice and 89% for DBA/1 mice was demonstrated when the sucrose in the liquid diet was replaced by an equicaloric (6% v/v) amount of ethanol. The ethanol-drinking isolated mice suffered a higher mortality rate than the grouped mice. When isolated mice were given a restricted amount of ethanol diet to match the consumption of the grouped mice, their death rate still remained higher. It was concluded that isolation increased the sensitivity to ethanol effects. The observed changes in the sensitivity to ethanol effects may have been mediated by the known isolation-induced changes in the levels of brain amines and corticosterone. Generally, DBA/1 mice were more susceptible to chronic ethanol than C57BL/10, and the young more susceptible than the adults.


Differential potency of taste and audiovisual stimuli in the conditioning of morphine withdrawal in rats. Morphine dependent rats that underwent naloxone-precipitated withdrawal in the presence of both gustatory and audiovisual stimuli subsequently avoided the taste cue, but not the audiovisual one. All environmental stimuli do not associate equally with withdrawal in the rat. The role of stimulus factors should be investigated in other forms of narcotic-related conditioning.


Circadian rhythm of corticosterone in mice: the effect of chronic consumption of alcohol. The effect of chronic consumption of alcohol on the circadian variations of the plasma corticosterone investigated in DBA/2J male mice. After 15 weeks of alcohol consumption (3.8%w/v for the first week and 7.5% for subsequent weeks) the alcohol groups exhibited a flattened circadian corticosterone curve, the level being intermediate between the peak and trough values of the water control groups. The diurnal patterns of food and liquid consumption were still present at the 10th week of alcohol treatment in the alcohol groups, although the  absolute amount of food and liquid consumed at each of the 6-h intervals was somewhat different between the alcohol and water groups. The blood alcohol showed a peak at early morning with the mean of 100 mg/100 ml, but the levels of alcohol during the remaining periods were remarkably stable, the means ranging from 30 to 46 mg/100 ml. Chronic consumption of alcohol, even relatively low concentrations, appears to affect the neural sites in the CNS controlling the circadian rhythm of ACTH release.


Effect of cortisone, aldosterone and nialamide on "amphetamine stereotypies" and brain methamphetamine levels of adrenalectomized rats. Cortisone, aldosterone or nialamide was administered to adrenalectomized or sham-operated rats for 7 days, and methamphetamine was injected 24 hrs after the last injection of these compounds. Stereotyped head movement and licking activity were scored 5 min, 30 min and 60 min after methamphetamine injection and, in parallel brain methamphetamine levels in similarly treated rats were measured 5 min, 30 min and 60 min after the methamphetamine injection. Adrenalectomy depressed stereotyped head movements but enhanced the brain amphetamine accumulation. Nialamide but not the hormones further increased the amphetamine accumulation in adrenalectomized rats. No drugs had any effect on the amphetamine-induced head movement suppressed by adrenalectomy.


Pigeon breeder's lung.

Pigeon breeder's lung. In the acute form of pigeon breeder's lung, chest X-rays often reveal micronodular changes, but 7 of the 16 patients examined showed no changes. In the subacute form, chest X-ray changes are exceedingly rare: they occurred in only 1 of the 17 patients. Pigeon breeder's lung differs from the condition known as bird breeder's lung in that it turns much less often into a chronic condition (pulmonary fibrosis and emphysema).


The effect of pyrithioxine and pyridoxine on individual behavior, social interactions, and learning in rats malnourished in early postnatal life. Low protein (LP) or low calorie (LC) dietary regimens were applied in early postnatal life(1st-40th day of life) in male rats. After nutritional rehabilitation, open-field behavior in larger more illuminated boxes (HI, high intensity stimulus), and smaller, less illuminated boxes (HI, high intensity stimulus), and smaller, less illuminated boxes (HI, high intensity stimulus), and smaller, less illuminated boxes (HI, high intensity stimulus), dyadic interactions, and learning ability were investigated in these animals as adults (between the 200th to 300th day of life). LP malnutrition induced an increase of open-field activity with features of sterotypy both in LI and HI situations, an increase number of intersignal reactions during learning procedures without changes in other registered criteria of learning ability (latency, number of correct responses), and an increase of aggressive behavior in pair interaction. LC rats revealed only significant inhibition in LI--open-field activity and a slightly increased number in intersignal reactions during avoidance learning. With the aim of preventing previously described long-term deviations in early malnourished rats, some groups of animals with the above-mentioned early calorie or protein deficits were treated with pyrithioxine (Encephabol Merck) or pyridoxine in 10 doses of 40 mg/kg i.p. administered in the period when nutritional rehabilitation was carried out (between the 40th--50th day of life). The treatment with pyrithioxine reduced significantly behavioral disturbances in adult LP rats except the increase of intersignal reactions which was even potentiated. Pyridoxine was less effective but normalized the increase number of intersignal reactions both in LP and LC rats. The effect of pyridoxine of adult LC rats was interesting. There was significant improvement in all registered parameters of avoidance learning and a significant increase of sexual acts was recorded.


Decrease of serotonin N-acetyltransferase activity in rat pineal organs after treatment with prostaglandin synthesis inhibitor indomethacin. Administration of indomethacin to rats abolished the cyclic AMP dependent, dark induced rise in serotonin N-acetyltransferase, presumably by inhibiting prostaglandin synthesis.


Production of prostaglandin Falpha in ewes following luteal regression induced with a prostaglandin analogue, Estrumate (cloprostenol; I.C.I. 80996). A single injection (100 mug i.m.) of Estrumate (I.C.I. 80996) was used to induce luteal regression on day 8 of the estrous cycle in 3 sheep. Progesterone levels in the utero-ovarian vein and femoral artery had fallen within 6 h to less than 50% of the concentrations seen before injection of the analogue. Luteolysis was not associated with endogenous production of PGF. The concentration of PGF in the uteroovarian vein began to increase 27-39 h after the administration of Estrumate, reaching a mean maximum concentration of 1455pg/ml 48 h after Estrumate. The mean concentration of PGF in the utero-ovarian vein between 36-69 h after Estrumate was significantly greater than during the 24 h before Estrumate (control period) or during the 0-30 h immediately after injection (both P less than 0.001). The maximum secretion of estradiol and the pre-ovulatory LH peak occurred during the period of elevated PGF concentrations in the utero-ovarian veins. The possible importance of endogenous PGF production at this time is discussed.


In vitro production of prostaglandins E and F by the guinea pig ovarian tissue. The ability of guinea pig ovarian tissue to biosynthesize prostaglandins E and F from endogenous precursors has been investigated in vitro. Estimations of prostaglandins were carried out using a sensitive radioimmuno assay during seven days preceding, and up to one day, following oestrous. Prostaglandins E and F were present in the ovarian tissue throughout the period investigated. Prostaglandin concentrations in samples incubated without enzymic inhibition were significantly higher than in samples incubated after enzymic inhibition with ethanol. This indicates that guinea pig ovarian tissue is able to synthesize prostaglandins from endogenous precursors.


Forest to pasture: development or destruction? The expansion of artifical or planted pastures (repasto) at the expense of both natural forest and cropland has brought major changes to the landscape and economy of Central America in recent years. On the pioneer fronts at the forest margin agriculture is commonly a transitory stage between forest felling and the establishment of permanent pasture. In the past 15 years the area in planted pasture and the total numbers of beef cattle have nearly doubled in several Central American countries while the per capita consumption of beef has actually declined. The "grassland revolution" that is occuring in Central America and Panama has been based almost exclusively on grass species of African origin which have in most cases been introduced into Central America only in this century. Of these jaragua, guinea, pangola, and kikuyu are the most important. The ecologic consequences of the conversion of forest to pasture are little understood. Intensive management practices, including the application of high-cost and scarce fertilizers and carefully-manged rotational grazing, will probably be necessary to sustain the productivity of these lands under conditions of tropical climate.


Cervical mucus changes in infertile ewes previously exposed to oestrogenic subterranean clover. Characteristics of cervical mucus were compared between groups of ewes which had previously grazed for long periods on either potently oestrogenic clover pasture (group A), mildly oestrogenic pasture (group B) or non-oestrogenic pasture (group C). No phytooestrogens were fed at the time of the study. The chloride content of mucus from group A ewes was significantly decreased, on a dry weight basis. The motility and survival of ram spermatozoa appeared similar in cervical mucus from groups A and B. There was no statistically significant difference between groups of ewes in either the total wet weight of mucus, the proportion of dry weight, or the degree of arborisation of mucus, at the time of oestrus. Mucus from group A ewes had a significantly decreased spinbarkheit, and a decreased ability to orientate sperum migration, when compared with group B. The group B ewes had a significantly smaller spinbarkheit than group C. Measurement of cervical mucus spinbarkheit offers promise as a diagnostic test for permanent clover infertility.


The urethral pressure profile in female patients with meatal stenosis. Urethral pressure profiles were recorded in a urodynamic study of 70 female patients with meatal stenosis before and after meatotomy. The study did not reveal any significant difference in the amplitude or contour, whereas a significantly longer urethra was observed in these patients than in normal women. Accordingly, the urethral pressure profile is not a diagnostic aid in deciding whether meatal stenosis is of pathogenic importance in the development of lower urinary-tract symptoms in women.


[Disorders of the emotional behavior during cerebral lesions]. A systematic study of the emotional behavior of right and left brain-damage patients was conducted on large groups of patients with unilateral cerebral lesions. The incidence of "catastrophic reactions" was found to be significantly higher among the left brain-damaged patients, whereas "indifference reactions" were significantly more frequent in the group with right-sided lesions. The depressive-catastrophic reactions of left brain-damaged patients seemed due chiefly to marked difficulties in verbal expression, while the indifference reactions of the right brain-damaged patients were highly correlated with neglect phenomena for the opposite half of the body and of space. The meaning of the emotional reaction shown by left brain-damaged patients seems easy to understand, if we consider that these subjects are affected by aphasia and by a paresis of the right hand. More surprising and paradoxical is, on the contrary, the emotional behavior of the right brain-damaged patients. The hypothesis is advanced that the indifference of these subjects may be only apparent, and due to a strong need of denying illness. The prevalence of denial of illness among the right brain-damaged patients could be due to the "non verbal" functional organization which is supposed to be characteristic of this half of the brain.


Diamond-Blackfan syndrome: lymphocyte-mediated suppression of erythropoiesis. Peripheral blood lymphocytes from six patients with congenital hypoplastic anemia suppressed erythroid cell formation by normal human bone marrow cells in response to erythropoietin in vitro. The results suggest that the anemia in these children has an immunologic basis.


[Review of the incidence of antibodies to various serological groups of the species Leptospira interrogans in a number of farm animals in the Netherlands (author's transl)]. Report on the results of serological studies on the species Leptospira interrogans in cattle (19,607), swine (6,348), dogs (182) and horses (88) from the Netherlands during the period from 1969 to 1974. Living cultures of the serotypes of pomona, icterohaemorrhagiae, canicola, guidae (Tarassovi serological group), grippotyphosa and sejroe were used as antigen in the micro-agglutination test. The numerical findings showed that antibodies to serotypes of the species Leptospira interrogans were present in 7.67 per cent of the cattle, 22.21 per cent of the pigs, 36.81 per cent of the dogs and 92.05 per cent of the horses studied. Infection with the serotypes of icterohaemorrhagiae and grippotyphosa was most common in cattle and horses, icterohaemorrhagiae and tarassovi were most common in swine and icterohaemorrhagiae and canicola were most common in dogs. The presence of pomona is not a factor in the Netherlands. In view of statistical findings, grippotyphosa infections in cattle may be assumed to result in abortion.


[Characteristics of different sublines of VNK-21 cells]. Comparative cytological, histological, histochemical and partially karyological studies were carried out on 8 batches of BHK-21 cell kept at different conditons. It was found that cells adapted to suspension culture differed from monolayer culture cells both morphologically and cytochemically. Suspension cells transmitted into monolayer culture displayed smaller size, changeable form, increased glycogen accumulation, decreased enzymatic activity (acid phosphatase, succinate dehydrogenase), their karyotypes tending to hypodiploidy.


Hemolytic disease of the newborn due to anti-N. A Negro woman of phenotype M+, N-, S-, s-, U+, who produced anti-N, is described. The antibody in her serum caused hemolytic disease of the newborn in her M+, N+ infant.


Anti-Dia and the Dia blood group. Antigen found in an Austrian family. In an Austrian family the Dia antigen was discovered by causing hemolytic disease of the newborn. No mongoloid admixture has so far been detected. No linkage to other blood groups, serum groups or red cell enzymes could be found.


[Primordial structures of feminine maturation as reflected in fairy tales. Contribution to the anthropology of sexuality]. Two maturation fairy-tales with a high resonance effect are analysed. The method of interpretation was that of depth psychology, with observation of results of modern biological-anthropological research. The interpretation revealed 'hymenal' or retarding primary structures in the young female subject. These structures postpone the onset of sexual desires to the time after the defloration.


[Participation of the lungs in the formation of an immune response]. The process of accumulation of the antibody-forming cells (AFC) in the lungs after vaccination of rabbits with the typhoid antigen was studied. No AFC was revealed in the lungs after single aerosol and subcutaneous immunization; double administration of the antigen through the respiratory tracts was followed by significant AFC accumulation in the lung tissue; individual antibody-synthesizing cells were found in the lung after subcutaneous application. Revaccination was followed by intensification of the immune response of the lung in both methods of vaccination; the content of antibody-producing cells in the aerosol-immunized rabbits was much greater than in the animals vaccinated subcutaneously. This indicated a definite role of the lungs in the immunogenesis in the case of the antigen administration through the respiratory tract.


Congenital contractural arachnodactyly.

Congenital contractural arachnodactyly. Five cases of congenital contractural arachnodactyly (CCA) are reported. Three belong to the one family. CCA has often been mistaken for Marfan's disease and arthrogrypois multiplex. Because CCA has a more favourable prognosis, it is very important to be able to recognize this syndrome.


The genetics and pathology of discrete subaortic stenosis in the Newfoundland dog. Breeding experiments confirm that discrete subaortic stenosis (SAS) in Newfoundland dogs is a specific inherited trait. Specificity of the morphogenetic abnormality is not complete, however, since matings between Newfoundlands with SAS occasionally produced pups with valvular and subvalvular pulmonic stenosis as well as SAS. The spectrum of severity of SAS ranged from a subclinical forme fruste to a severe form causing death before maturity. Well-developed subvalvular stenotic rings consisted of a base of loosely arranged fibrous connective tissue and a subendocardial region of cartilagenous tissue. Severely affected dogs, some of which died suddenly, had foci of necrosis and fibrosis in the left ventricular myocardium, associated with thickening of the intramural coronary arteries. The lesions of SAS were not found in dogs before 3 weeks of age, and the mildest form was seen only in dogs between 3 and 12 weeks of age, suggesting that SAS is not a true congenital defect but develops postnatally. It is hypothesized that the fibrocartilagenous ring of SAS is derived from persistent embryonal endocardial tissue which retains its proliferative capacity and has chondrogenic potential for some time after birth. The results of breeding experiments were not consistent with any simple genetic hypothesis, and indicate that SAS is inherited as a polygenic trait or as an autosomal dominant trait with modifiers.


Immunocytologic losses of isoantigens A, B, and H (O) in the endometrium and brain. Formalin-fixed tissues from 100 endometria and 50 brains were grouped and studied by the technic of mixed-cell agglutination reaction (MCAR) for studying isoantigens A, B, and H (O). MCAR's were negative in all 45 of the endometria from subjects with endometrial carcinomas, where as MCAR's were positive in the epithelium of endometrial glands of the remaining 55 subjects (cyclic phases of endometrium and benign lesions). MCAR's were negative in all benign and malignant brain tumors and normal brain tissues used in this study. In view of the present findings and in the light of previous observations, the isonatigen loss in adenocarcinomas is greater than such losses in other types of malignancies so far studied. The cause of the negative MCAR's in normal brain tissue and brain tumours is not known, but they may be attributable to lack of isoantigens in normal brain tissue.


Effects of carbenoxolone Na on acute and chronic gastric ulcer models in experimental animals. Effects of carbenoxolone Na on acute or chronic types of gastric lesions or ulcer models produced in rats, guinea pigs, or dogs were studied. Carbenoxolone Na, given either orally or intraperitoneally, produced a significant inhibition of stress-induced gastric lesions in intact or in pylorus-ligated rats. Acetylsalicylic acid (ASA)-induced or serotonin-induced gastric lesions in rats were also inhibited significantly by pretreatment with the drug. However, carbenoxolone Na did not affect the development of Shay ulceration in rats even though the peptic activity in gastric juices was markedly reduced by the drug. Histamine-induced gastric lesions in guinea pigs were not prevented by pretreatment with carbenoxolone Na. Although carbenoxolone Na, given for 10-20 days, did not promote the healing of stress-induced gastric lesions and acetic acid gastric jlcers in rats, it significantly accelerated the healing of chronic gastric ulcer produced in dogs by 3 weeks' treatment. Carbenoxolone Na prevented the acid back-diffusion caused by ASA without any influence on Na+ efflux in pylorus-ligated rats.


Sampling the fetoplacental circulation. I. The pregnant ovine. Intrauterine endoscopy (fetoscopy) was successfully accomplished in 26 specially prepared pregnant sheep at 85 to 110 days' gestational age, with the use of a conventional 2.7 mm. Hopkins straightforward wide-angle telescope (Stortz 27018A) encased in a prototype 3.1 mm. eccentric cannula which incorporates a 25 gauge needle channel. Fetal blood was aspirated in 12 lambs. Twenty-five animals were followed throughout the remainder of pregnancy: nine (36 per cent) aborted within three days-three weeks after endoscopy. The pregnant ovine appeared to be a useful experimental animal model for the development of an intrauterine endoscopy technique.


Light and electron microscopic study of early lesions in angiomatosis retinae. Multiple early untreated lesions of angiomatosis retinae in the peripheral retina and optic disk were discovered after death in both eyes of a patient with von Hippel-Lindau disease. Each of the four peripheral angiomas was studied by a different technique: routine light microscopy of serial histologic sections, electron microscopy, flat mount and trypsin digestion, and frozen section with special stains for lipid. The large capillaries comprising the core of each angioma displayed normal endothelium, basement membrane, and pericytes. Thus, capillary hemangioma of the retina is a more correct histopathologic designation for the von Hippel angioma than terms such as hemangioblastoma or hemangioendothelioma. Interstitial cells separating the vascular channels of the angioma were identified as astrocytes that contain large lipid-filled cacuoles. Serial histologic sections demonstrated that both retinal and choroidal vessels communicate with vascular channels of the juxtapapillary capillary hemangiomas.


Recurrent tapioca melanoma of the iris and ciliary body treated with the argon laser. A 25-year-old white woman underwent iridocyclectomy for an iris tumor. Clinically, histologically, and ultrastructurally this tumor was the same as a recently reported tapioca melanoma of the iris. During the next four months, there were recurrent growths on the ciliary body that we treated with the argon laser through a three-mirror lens. No further recurrences of the tumor were noted three years later.


Chorioretinal anastomoses after argon laser photocoagulation. Communication between the retinal and the choroidal circulation occurred in one case of proliferative diabetic retinopathy and one case of Leber's disease (multiple miliary aneurysms) after argon laser photocoagulation treatment to eliminate abnormal retinal vasculature. In both instances rapid-sequence fluorescein angiography documented the choroidal origin of the anastomoses. Such anastomoses developed in areas where an excessively dense laser beam aimed at occluding retinal vessels presumably destroyed the underlying Bruch's membrane and permitted invasion of the retina by fibrovascular growth from the choriod. The occurrence of such growths is a potential complication of argon laser photocoagulation directly related to the power density used.


Isolation and characterization of adeno-associated viruses from bovine adenovirus types 1 and 2. Hemagglutinating DNA viruses of 20 nm diameter were isolated from bovine adenovirus types 1 and 2. The isolates were heat stable, chloroform resistant, and defective. Their densities were 1.38 to 1.39 g/cm3, and they were found to be serologically identical to the bovine adeno-associated virus strain X7. A partial antigenic relationship was found between these and the canine adeno-associated virus.


Cyclic ultrastructural changes in ewe uterine tube (oviduct) infundibular epithelium. Ultrastructural features of the uterine tube (oviduct) infundibulum of ewes have been studied, with special reference to cyclic changes in the ciliated and the secretory cells. Tissue from the uterine tube infundibulum was taken from 12 Rambouillet crossbreed ewes which were killed at intervals (days 1 (or estrus), 3, 9, 10, 12, and 16) throughout the estrous cycle. The presence of cilia was demonstrated throughout the estrous cycle, and true degeneration or loss of cilia was not apparent at any phase of the cycle. Presence of fibrous granules, which are supposedly related to basal body replication, was demonstrated in the apical cytoplasm of ciliated cells on day 1 of the estrous cycle. Small ciliary buds were especially present on day 1, indicating active formation of cilia during the follicular phase of the cycle. The presence of fibrous granules, basal bodies, and ciliary buds at estrus indicates that ciliogenesis in the ewe uterine tube is stimulated by high levels of endogenous estrogen. Rootlets were observed both during the follicular and the luteal phases of the cycle. The rootlets were about 1 mum long, and their fine structure indicates that they might function as anchoring structures for the motile cilia. The most striking feature during estrus was the occurrence of glycogen granules in the cytoplasm of ciliated and secretory cells. These granules were in the apical cytoplasm and basal region of some epithelial cells. They were minimal or absent during the luteal phase of the estrous cycle. The presence of electron-dense glycogen particles was clearly demonstrated within basal bodies. Possibly the glycogen within the basal bodies functions as a source of energy for ciliary movement and the cytoplasmic glycogen as nourishment for the ovum. The secretory cells also showed characteristic cytologic changes which were correlated with the phase of the estrous cycle. Maximal secretory cell differentiation was apparent during the follicular phase, at which time these cells were characterized by well-developed rough endoplasmic reticulum, numerous ribosomes, and secretory granules of varied size, shape, and density. A most remarkable feature of the granules was their membranous structure, consisting of concentric lamellae of equal dimensions. Typical extrusion of secretory granules into the tubal lumen was apparent during the follicular and the luteal phases of the estrous cycle. Cytoplasmic projections containing nuclei protruded into the tubal lumen and some were free in the lumen, especially during the luteal phase of the estrous cycle. The presence of a well-developed endoplasmic reticulum and numerous secretory granules during estrus indicate that secretion in the ewe uterine tube is presumably under the control of circulating high plasma concentrations of estrogen.


Clinical pharmacokinetics of sisomicin: dosage schedules in renal-impaired patients. The pharmacokinetics of intravenously administered sisomicin were studied in 33 patients with normal renal function and different degrees of renal impairment. In all patients, the serum disappearance of sisomicin, once distribution equilibrium had been achieved, followed first-order kinetics and percentage of hourly loss from serum decreased proportionally with decreasing renal function. Half-lives averaged 2.06 h in normal subjects (endogenous creatinine clearance greater than 80 ml/min per 1.73 m(2)) and reached 35.3 h in a virtually anephric subject. Linear relationships were defined between sisomicin serum half-life and the reciprocal of the endogenous creatinine clearance and serum creatinine concentration. The latter relationship indicates that the half-life of sisomicin may be approximated as twice the serum creatinine concentration and may be used for dosage adjustment in renal-impaired patients. Prediction of the extent of sisomicin removal by hemodialysis may be made from the relationship between the dialyzate of sisomicin and that of creatinine and blood urea nitrogen. Dosage schedules and methods of administration compatible with the pharmacokinetic properties of the antibiotic are finally proposed.


Jaundice and bilirubin levels in Greek children with favism. Jaundice and bilirubin levels varied widely in 85 children with favism. Low bilirubin levels (less than 2 mg/100 ml) with clinically undetectable jaundice were seen in 34 (40%), mild jaundice in 32 (38%), moderate in 16 (19%), and severe (greater than 8 mg/100 ml) in 3 (4%). Bilirubin levels were unrelated to the severity of anaemia or to reticulocytosis. The absence of bilirubinaemia and jaundice in a high proportion of the patients was attributed to the ability of the liver to conjugate large amounts of bilirubin. The extreme bilirubinaemia and jaundice observed in a minority of cases was attributed to the existence of an additional hereditary factor affecting the liver.


Gamma-aminobutyric acid: selective inhibition of electrically induced head-turning following intracaudate administration. GABA selectively antagonized head-turning elicited by electrical stimulation of the caudate nucleus in the rat when applied directly to the site of stimulation utilizing an electrode-cannula. Pretreatment with picrotoxin, a GABA antagonist, prevented the action of GABA on the head-turn response. This report provides in vivo evidence that GABA may play an inhibitory role in basal ganglia function. The findings are relevant to basal ganglia disorders in man.


Induction of lactation in virgin goats by perhenazine. Chronic implantation of a small amount of perphenazine into the median eminence (M.E.) of virgin female goats caused mammary development and artificial induction of lactation. Sham implantations in the M.E. and implantation of perphenazine in other sites of the hypothalamus were ineffective.


Angiosarcoma of the heart. A case study. Angiosarcoma of the heart is a rare neoplasm. The clinical manifestations are surprisingly consistent, but the diagnosis is rarely made before death. A patient with such a tumor was described here and the current literature was reviewed. Venous angiography and pericardiectomy are recommended in patients with hemorrhagic pericardial tamponade when the cause is unclear.


Frequency of sexual dysfunction in a general gynecological clinic: an epidemiological approach. Epidemiological data on female sexual functioning are presented. Fifty-nine 30- to 39-year-old Black women attending a gynecology clinic at University Hospitals of Cleveland for nonsexual complants were interviewed to determine the frequency of sexual dysfunction. Seventeen percent had difficulty achieving orgasm in partner sexual behavior. Most of these women had prior history of adequate sexual functioning; 5% had never been orgasmic. Ten of the 49 women who had no orgasmic difficulty were not satisfied with their sexual relationships. The relative risk of sexual dysfunction was 5 times greater in women who had undergone pelvic surgery.


Group therapy for nonorgasmic women: two age levels. A group method of therapy was used for the treatment of primary orgasmic dysfunction ten women below and ten women above 35 years of age to determine the appropriateness of this therapy for younger and older females. The treatment combined a Masters and Johnson style of behavioral therapy and self-stimulation therapy. At termination, 70% of the younger women were orgasmic, with 80% orgasmic at 6-month follow-up. Zero percent was coitally orgasmic. Forty percent of the older women were orgasmic at therapy termination, with 60% orgasmic at 6-month follow-up. One women was orgasmic during intercourse. There was general enhancement of the sexual relationship in all participating couples. Group therapy may be less successful for older women, who may be more successfully treated individually.


An inventory for the measurement of female sexual arousability: the SAI. This report describes the development of a self-report Sexual Arousability Inventory (SAI) for women. Sexual arousability was defined as the sum of a respondent's ratings of 28 erotic experience along a 7-point Likert arousal dimension. Multiple-regression and factor analyses were used to select valid items from a 131-item pool and build in factorial purity. The SAI has concurrent validity with respect to sexual experience, activity, and satisfaction, and discriminates between clinical and normal populations. In addition, the SAI is easy to administer and score, may be used with single, married, or lesbian women, is available with norms and in alternate forms, and possesses exceptional internal consistency. Although the SAI was designed primarily for clinical use, the construct it measures may have theoretical significance in future research.


Isolation of substances from human vaginal secretions previously shown to be sex attractant pheromones in higher primates. Fractionation of vaginal secretions from sexually human females by partitioning and chromatographic procedures demonstrates the presence of short-chain fatty acids seen as crucial pheromonal components in previous higher primate studies. In addition, the appearance of these short-chain fatty acids suggests a possible correlation of the rise and fall of hormone levels during the female menstrual cycle.


The synthesis of polyadenylic acid-containing ribonucleic acid by isolated mitochondria from Ehrlich ascites cells. The synthesis of poly(A)-containing RNA by isolated mitochondria from Ehrlich ascites cells was studied. Isolated mitochondria incorporate [3H]AMP or [3H]UTP into an RNA species that adsorbs on oligo (dT)-cellulose columns or Millipore filters. Hydrolysis of the poly(A)-containing RNA with pancreatic and T1 ribonucleases released a poly(A) sequence that had an electrophoretic mobility slightly faster than 4SE. In comparison, ascites-cell cytosolic poly(A)-containing RNA had a poly(A) tail that had an electrophoretic mobility of about 7SE. Sensitivity of the incorporation of [3H]AMP into poly(A)-containing RNA to ethidium bromide and to atractyloside and lack of sensitivity to immobilized ribonuclease added to the mitochondria after incubation indicated that the site of incorporation was mitochondrial. The poly(A)-containing RNA sedimented with a peak of about 18S, with much material of higher s value. After denaturation at 70 degrees C for 5 min the poly(A)-containing RNA separated into two components of 12S and 16S on a 5-20% (w/v) sucrose density gradient at 4 degrees C, or at 4 degrees and 25 degrees C in the presence of formaldehyde. Poly(A)-containing RNA synthesized in the presence of ethidium bromide sedimented at 5-10S in a 15-33% (w/v) sucrose density gradient at 24 degrees C. The poly(A) tail of this RNA was smaller than that synthesized in the absence of ethidium bromide. The size of the poly(A)-containing RNA (approx. 1300 nucleotides) is about the length necessary for that of mRNA species for the products of mitochondrial protein synthesis observed by ourselves and others.


Trace concentrations of halothane in human breast milk. Halothane concentrations of 2 p.p.m. have been found in the breast milk of a lactating, practising anaesthetist. This concentration was consistent with the operating theatre environment. The authors fell that, in spite of the limited scope of the study, this is an additional reason for the elimination of waste anaesthetic agents from the operating theatre.


The cytotoxicity of plasma from patients with acute hepatic failure to isolated rabbit hepatocytes. The cytotoxicity of plasma from patients with various types of liver disease to rabbit hepatocytes maintained in vitro has been investigated using a microcytotoxicity assay system. Plasma from patients with fulminant hepatic failure and uncomplicated viral hepatitis showed significant cytotoxicity compared to controls. The cytotoxic effect of plasma from patients with fulminant hepatic failure was reduced by charcoal haemoperfusion, or heating and dialysis. The bile acids chenodeoxycholic acid and lithocholic acid were cytotoxic when added to control human plasma. Such toxic factors may be responsible for the delay in liver regeneration often seen in patients with fulminant hepatic failure.


Blood leucocyte enzymes. III. Diurnal rhythm of activity in isolated lymphocytes of normal subjects and chronic lymphatic leukaemia patients. The activities of glucose-6-phosphate dehydrogenase (D-glucose-6-phosphate: NADP oxidoreductase, G6PD), 6 phosphate glucono dehydrogenase (6 phospho-D-gluconate: NADP oxidoreductase, 6PGD) lactate dehydrogenase (D-lactate: NAD oxidoreductase, LDH), glutamate oxaloacetate transaminase (L-aspartate: 2-oxo-glutarate aminotransferase, GOT) and hexokinase (ATP: D-hexo-6-phosphotrans-ferase, Hx) were measured over 24 h in isolated lymphocytes of normal subjects and in white cells of patients with chronic lymphatic leukaemia (CLL). The activitty patterns of all enzymes in the normal lymphocytes were similar. A computed pattern of all the results exhibited a circadian rhythm of activity with the highest level at 16.00 hours. The oscillations in the activities of the same enzymes in the CLL cells differed among the patients, although all the enzymes of the same individual showed a similar diurnal rhythmic pattern. All peaks in this group appeared between 20.00 and 08.00 hours. The possible importance of these observations in setting up therapeutic schedules was raised.


Corneal neovascularization treated with argon laser. The argon laser set at 50 mum, 100-150 mW, o-2 s occluded corneal blood vessels in pigmented Dutch rabbits provided the corneal responsible for inducing vascularization was inactive. After arterial treatment with the argon laser retrograde flow in untreated veins was demonstrated by fluorescein angiography. Therefore all corneal new vessels should be treated, not just arteries. Minimal iris damage complicated the laser therapy, but this was not thought necessarily to contraindicate the use of the argon laser to treat corneal blood vessels in man. The iris damage was associated with outpouring of aqueous from the ciliary processes, and it took up to a week for the blood-aqueous barrier to return to its normal state.


[14C]leucine incorporation into proteins of pancreatic juice in rats fed soya-bean flour. 1. The effect of a diet containing a trypsin inhibitor on the incorporation of radioactively labelled leucine into the pancreatic proteins secreted during stimulation with cholecystokinin-pancreozymin (CCK) was studied in rats. 2. The total output of protein was significantly greater in the rats given raw soya-bean flour (RSF) compared with those given heat-inactivated soya-bean flour (HSF) (controls) in response to the sub- and supramaximal stimulation with CCK, but similar responses were obtained to maximal stimulation with CCK. Total protein output decreased continuously with time after reaching peak values at 90--120 min after the start of stimulation with CCK. 3. The total output of radioactively labelled protein in RSF-fed rats was not different from that of the controls with sub- and supramaximal dose rats of CCK, but was significantly lower than that of the controls in response to the dose rate of CCK which produced maximal rates of pancreatic secretion. 4. The specific activity of radioactively labelled protein increased continuously, while the output attained a constant rate during stimulation with all doses of CCK. 5. We concluded that feeding the trypsin inhibitor-containing diet led to increased secretion of stored pancreatic protein, while secretion of newly synthesized protein was not altered. During the course of prolonged stimulation with CCK, irrespective of diet, there was increasing secretion of the newly synthesized protein compared with the pre-existing stored proteins of the pancreas, it was unable to compensate for the decreased secretion of pre-formed protein.


Properties of crystalline reduced nicotinamide adenine dinucleotide phosphate-adrenodoxin reductase from bovine adrenocortical mitochonria. I. Physicochemical properties of holo- and apo-NADPH-adrenodoxin reductase and interaction between non-heme iron proteins and the reductase. A crystalline NADPH-adrenodoxin reductase was obtained from bovine adrenocortical mitochondria and its properties were investigated. Its molecular weights and isoelectric point were estimated to be 51 000 and 5.4, respectively. Amino acid and sugar contents and the interaction between the apo-reductase and flavin of NADPH-adrenodoxin reductase were investigated. Formation of a complex of bovine NADPH-adrenodoxin reductase with adrenodoxin, its apoadrenodoxin, or other non-heme iron proteins caused quenching of fluorescence of the tryptophanyl residue and bound FAD of the NADPH-adrenodoxin reductase. The results obatined suggest that adrenodoxin and apoadrenodoxin bind functionally to a site close to the tryptophanyl residue and the bound FAD of the reductase. The circular dichroism spectrum of oxidized NADPH-adrenodoxin reductase was measured in the ultraviolet and visible regions. This spectrum showed negative absorption in the visible region and was not appreciably influenced in either the ultraviolet or visible region by formation of a complex with adrenodoxin or apoadrenodoxin.


Nucleic acid polymerizing enzymes in developing Strongylocentrotus franciscanus embryos. DNA-dependent RNA polymerase, DNA-Dependent DNA polymerase, and terminal riboadenylate transferase (TRT) activities have been measured after DEAE-Sephadex chromatography of whole cell extracts prepared from eggs and staged embryos of the urchin, Stronglyocentrotus franciscanus. Activity of each of these three polymerase classes is present in the egg, and the total activity per embryo is constant throughout embryogenesis to the pluteus stage (approximately 1000 cells). Thus the egg appears to contain sufficient DNA polymerase, RNA polymerase, and TRT TRT for embryogenesis. The increases in the synthesis of DNA, RNA and polyadenylated RNA tracts observed after fertilization must be due to the activation of the preexisting egg enzymes. Separation of the egg into nucleate and anucleate halves demonstrates that the RNA polymerases are not restricted to the egg nucleus. During development, the enzymes become progressively more associated with the cell nucleus. The egg extracts contain low activities (approximately 6% total) of RNA polymerase II as measured by sensitivity to alpha-amanitin. This is confirmed by resolution of the RNA polymerase forms I, II, and III by gradient sievorptive elution on DEAE-Sephadex. Later stage embryos contain more nearly equal activities of RNA polymerase, I, II, and III, although the total RNA polymerase activity per embryo is not changed. Additionally, two chromatographicallly distinct species of RNA polymerase III are detected, one of which is observed only in later stages. Thus interconversion of enzymes via addition of new subunits or coordinate synthesis and loss of enzyme species must occur.


Effect of polylysine on the activation of prothrombin. Polylysine substitutes for calcium ions and factor V in the factor Xa catalyzed activation of prothrombin. Polylysine has been demonstrated to dramatically accelerate the rate of the factor Xa catalyzed activation of both prothrombin and prethrombin 1. Under the present experimental conditions (pH 8.0, 23 C), no detectable activation of prothrombin or prethrombin 1 occurs with either factor Xa or polylysine alone. The activation of prethrombin 2, the direct precursor of alpha-thrombin, by factor Xa is not stimulated by polylysine. The activation of either prothrombin or prethrombin 1 by factor Xa in the presence of polylysine is partially inhibited by the presence of 5 mM CaCl2. Electrophoretic analysis in sodium dodecyl sulfate showed that the products that were formed in the above activation system comigrated with  the reaction products derived from prothrombin activated by factor Xa in the presence of calcium ions and phospholipid. It is suggested that polylysine stimulates the factor Xa-catalyzes activation of prothrombin by replacing the combination of calcium ions and factor V.


Extended RNA synthesis in isolated nuclei from rat pituitary tumor cells. Nuclei of GH3 cells, isolated by detergent lysis, synthesized RNA for an extended period at 29 degrees C in the presence of rat liver ribonuclease inhibitor (RI). Extended RNA synthesis was dependent upon the presence of RI. Sucrose gradient sedimentation analysis of the cell-free reaction products showed that RNAs ranging from 4 S to greater than 28 S were synthesized. Further characterization of the RNA products was made by examining the sensitivity of synthesis to alpha-amanitin and actinomycin D as well as by oligo(dT)-cellulose binding properties. Evidence was obtained that RNA polymerases I, II, and III were functioning in isolated GH3 nuclei. Newly synthesized RNAs were found in both the nuclear pellet and postnuclear supernatant fractions. RNA polymerase I products remained associated with the nuclear pellet throughout a 60-min incubation period whereas RNAs synthesized by RNA polymerase III emerged rapidly into the supernatant fraction. RNA polymerase II products were distributed in both fractions and were found to contain poly(A). De novo poly(A) synthesis was demonstrated and found to be inhibited by cordvcepin triphosphate (3'-dATP). Supernatant RNAs synthesized by polymerase II contained a poly(A) segment of about 150 adenine residues; these transcripts sedimented heterogeneously with an apparent size distribution (under denaturing conditions) which was smaller than that of nuclear RNA polymerase II products and which resembled that of cellular mRNA. Qualitative differences in the nuclear and supernatant RNAs, the kinetics of appearance of the latter, and the differential effect of 3'-dATP on the extranuclear appearance of supernatant RNAs suggest that a process resembling nuclear-cytoplasmic RNA transport occurred in this cell-free nuclear system.


Purification of myosin translational control RNA and its interaction with myosin messenger RNA. Myosin messenger ribonucleoprotein-translational control ribonucleic acid (mRNP-tcNA) from myosin mRNPs found in embryonic chick muscle has been further purified by Dowex chromatography and, from a number of controls, it is suggested that this small RNA is not an artifact produced through the degradation of RNA during its isolation. This highly purified myosin mRNP-tcRNA is shown to have a molecular weight of 10 000 on formamide-acrylamide gels, and reacts stoichometrically (on a 1:1 mole ratio) with myosin mRNA. The stoichiometric interaction between myosin mRNA and myosin mRNP-tcRNP is demonstrated by ists ability to increase the nuclease resistance of the messenger, as well as inhibit its translation in a cell-free amino acid incorporating system.


Messenger ribonucleic acid metabolism in mammalian mitochondria. Discrete poly(adenylic acid) lacking messenger ribonucleic acid species associated with mitochondrial polysomes. The mRNA species released from mitochondrial polysomes prepared by the Mg2+ precipitation technique have been further characterized using various analytical techniques. Mitochondrial polysomes were dissociated by treatment with puromycin and chemically labeled with (3H) dimethyl sulfate. About 51% of steady-state mitochondrial mRNA bind to oligo(dT)-cellulose indicating the presence of poly(adenylic acid)(poly(A)) in this fraction. The poly(A)-containing mRNAs resolve into discrete bands of 9-16 Se, while the RNA fraction unable to bind to oligo(dT)-cellulose representing poly(A)-lacking mRNA contains 8-12 Se species. About 90% of poly(A) lacking RNA hybridizes with mitochondrial DNA and less than 7% hybridizes with nuclear DNA. The extent of hybridization of poly(A)-lacking RNA with mitochondrial DNA was not significantly affected by the presence of excess mitochondrial rRNA, cytoplasmic rRNA, or a tenfold concentration of poly(A)-containing RNA isolated from total mitochondrial RNA. Possible differences in sequence properties between poly(A)-containing and -lacking mitochondrial mRNAs were further verified using a solid phase-bound cDNA procedure. Poly(A)-containing mRNA released from mitochondrial polysomes shows over 85% sequance homology with oligo(dT)-cellulose-bound cDNA prepared against total mitochondrial poly(A)-lacking mitochondrial mRNA hybridizes with the cDNA providing direct evidence for the distinct sequence properties of the two mRNA species.


Conformation of immunoglobulin M. I. Characterization of anti-epsilon-1-dimethylamino-5-naphthalenesulfonyl-L-lysine immunoglobulin M antibodies from horse, pig, and shark. IgM antibodies specific for the fluorophore epsiolon-1-dimethylamino-5-naphthalenesulfonyl-L-lysine(DNS-lysine) were elicited in the horse and nurse shark by immunization with a DNS-lysine streptococcal conjugate; the antibodies were purified by specific adsorption with an immunoadsorbent followed by gel filtration to select the IgM class (molecular weight 900 000). About 90% of the equine anti-DNS was IgM.DNS-Lysine, when bound in the combining sites of a population of these anti-DNS IgM antigodies from horse and nurse shark, as well as from pig, exhibited a marked fluorescence enhancement and shift of the emission spectrum to shorter wavelengths compared with emission in aqueous solution; these results indicate that the environments of the anti-DNS combining sites of  this population were relatively hydrophovic. Approximately one-third of the ten possible combining sites in each of these anti-DNS IgM species bound DNS-lysine in this manner with an average intrinsic association constant (Ko) of greater than  10(6) M-1. Small differences were noted in binding behavior among the three species of antibodies. The enzymatic susceptibility of equine IgM was similar to that of human IgM. (Fab')2mu, Fab'mu, and Fabmu fragments were prepared following digestion with pepsin. These fragments could be clearly differentiated on the basis of molecular size. They bound DNS-lysine with the same affinity as intact IgM  and the DNS-lysine-fragment  complexes exhibited the same spectral properties as the parent IgM. It was concluded that the anti-dNs IgM antibodies from all three species, as well as the enzymatic fragments, were suitable for nanosecond depolarization studies which are reported in the accompanying paper Holowka, D.A., and Cathou, R.E. (1976), Biochemistry, the following papter in this issue.


Conformation of immunoglobulin M. 2. Nanosecond fluorescence depolarization analysis of segmental flexibility in anti-epsilon-l-dimethylamino-5-naphthalenesulfonyl-L-lysine anti-immunoglobulin from horse, pig, and shark. The rotational motions of immunoglobulin M (IgM) were investigated by the nanosecond fluorescence depolarization technique. The fluorophore epsilon-1-dimethylamino-5-naphthalenesulfonyl-L-lysine (DNS-lysine) was specifically bound in the combining sites of anti-DNS IgM antibodies from the horse, pig, and nurse shark. Fluorescence lifetime analysis showed the presence of a long lifetime component (21-27 ns) with antibodies from all three species. With the mammalian antibodies, the fluorophore appeared to be rigidly bound in the combining sites as judged by the presence of induced circular dichroism of DNS-lysine (equine antibodies) and single exponential anisotropy decay of the isolated Fabmu fragments (equine and porcine antibodies). The small amount of available purified nurse shark antibody did not allow preparation of fragments or induced circular dichroism measurements to directly determine rigidity of fluorophore binding. However, at least some of the hapten must have been rigidly bound since long rotational correlation times were measured for the shark DNS-lysine-anti-DNS complexes. When the emission anisotropy of the fluorophore-anti-DNS IgM complexes was measured as a function of time, it was found that all three antibody species exhibited restricted segmental flexibility in the nanosecond time range. Moreover, when the equine anti-DNS IgM was exposed to 1 M acetic acid for 1 h, the antibody underwent a conformational change which resulted in an increase in its overall flexibility. Comparison of the rotational correlation times of native equine IgM and of proteolytic fragments indicated that flexibility of IgM consists of either hindered rotation of the Fab'mu segment or a combination of at least two modes of motion: rotation of Fabmu and/or Fab'mu and bending of the entire (Fab')2mu region as a unit. Similar modes of flexibility also occur in native porcine IgM. In acid exposed equine IgM, the major contribution to depolarization is from independent rotation or wagging of the Fab'mu segments. Thus, acid apparently causes a conformational change in or near the Cmu2 domains. In contrast, flexibility in nurse shark IgM appears to involve only bending of (Fab')2mu as a unit. Our results suggest that segmental flexibility is an essential functional feature of all IgM antibodies and that control of this flexibility through domain interactions may play an important role in such conformationally sensitive functions as complement fixation.


Formation of delta1-acetoxytryptophan-62 in the oxidation of tryptophan-62 of hen egg-white lysozyme by N-bromosuccinimide in acetate buffer. The reaction of equimolar amounts of N-bromosuccinimide and hen egg-white lysozyme in acetate buffer, under the conditions of Hayashi et al. (Hayashi, K., Imoto, T., Funatsu, G., and Funatsu, M. (1965), J. Biochem. (Tokyo) 58, 227), yields a protein mixture that has a time-dependent 13C-NMR spectrum. The initial natural-abundance 13C-NMR spectrum indicates the presence of about equal amounts of [oxindolealanine-62]lysozyme and [delta1-acetoxytryptophan-62]lysozyme. The latter converts to [oxindolealanine-62]lysozyme with a half-life of about 2 days at 25 degrees C and pH 3.9. Two observations indicate that the source of the acetyl group of delta1-acetoxytryptophan-62 is the acetate buffer. First, the spectrum of a lysozyme sample treated with N-bromosuccinimide in the presence of [1-13C]acetate yields a very strong acetyl ester carbonyl resonance. The time dependence of the intensity of this resonance yields a half-life of 44 h for [delta1-acetoxytryptophan-62]lysozyme. Second, the initial natural-abundance 13C-NMR spectrum of a lysozyme sample treated with N-bromosuccinimide in the absence of acetate indicates essentially complete conversion of tryptophan-62 into oxindolealanine.


Mechanism of hydrolysis by serine proteases: direct determination of the pKa's of aspartyl-102 and aspartyl-194 in bovine trypsin using difference infrared spectroscopy. The pKa of aspartyl-102 in trypsin is shown to be 6.8+/-0.2 by difference infrared titration. All but 2.5 of the carboxyls in bovine trypsin were first modified with semicarbazide. The modified enzyme still retains full activity toward nonspecific substrates. The remaining free carboxyls include one equivalent each of Asp-102 and Asp-194. The absorbances associated with the C=O and C=O stretching modes at 1570 and 1710 cm-1 were used to monitor the proportion of ionized or protonated carboxyl present in the enzyme as a function of pD. The pKa of 6.8 was assigned to Asp-102 using copper ions that bind to trypsin between Asp-102 and histidine-57, so lowering the pKapp of Asp-102. The implication of this result for the ionization of the active site, and for the mechanism of serine proteases, is discussed. Asp-194 and the C terminus are shown to titrate with an average pKa of 2.9.


A spectroscopic technique for measuring slow rotational diffusion of macromolecules. 1: Preparation and properties of a triplet probe. Slow rotational diffusion may be investigated by measuring the decay of dichroism of flash-induced transient absorption changes of suitable probes. The preparation of the covalent ""triplets" probe eosin isothiocyanate is described together with investigations of spectroscopic properties of eosin-protein conjugates. Triplet state lifetimes of air-equilibrated solutions of eosin-protein conjugates are in the order of 10 mus, demonstrating that the probe is protected from oxygen quenching by the protein. Experiments with the sucrase-isomaltase complex from small intestine show that its enzymatic activity is little affected by binding up to 2 mol of eosin/mol of protein.


A spectroscopic technique for measuring slow rotational diffusion of macromolecules. 2: Determination of rotational correlation times of proteins in solution. Experiments have been performed to test the validity of measurements of slow rotational diffusion using eosin isothiocyanate as a probe. When eosin-protein conjugates are covalently bound to Sepharose 4B, the dichroism of the flash-induced absorption transient does not decay with time. The magnitude of the dichroism is smaller than for eosin in solid solution. These results, together with measurements with highly viscous solutions, demonstrate the existence of a rapid but restricted independent motion of the eosin probe. This causes a partial loss of pf dochroism leaving a residual dichroism whose time dependence is determined by the rotational motion of the protein. Rotational correlation times in the microsecond-millisecond time range have been determined for eosinprotein conjugates dissolved in glycerol-water mixtures of varying viscosity. The measured values are in good agreement with theoretical predictions and show the correct dependence on the viscosity of the medium and the size and state of association of the protein.


Isolation, physicochemical properties, and the macromolecular composition of the vitelline and fertilization envelopes from Xenopus laevis eggs. As a step toward defining in molecular terms the sperm-triggered block to polyspermy reaction established by the egg at fertilization, vitelline (VE) and fertilization (FE) envelopes were isolated from eggs of the Sounth African clawed toad Xenopus laevis and some of their physicochemical properties determined. Envelopes were isolated after lysis of the fertilized or unfertilized eggs by sieving techniques; isolated envelopes retained their in situ morphology as determined by electron microscopy. The isolated envelopes had different solubility properties and, in general, VE was more readily dissolved by aqueous solvents than FE, although both could be completely dissolved by detergents or chaotropic agents. Changes in envelope solubility correlated with the progression of the cortical reaction implicating a role for cortical granule material in modifying the solubility properties of the envelope. The VE and FE were composed of protein and carbohydrate with no lipid components detected. As determined by immunodiffusion experiments, the FE contained the same antigens as the VE plus components derived from the cortical granules and the innermost jelly layer, J. The macromolecular composition of the envelopes was determined by sodium dodecyl sulfate gel electrophoresis. The VE contained at least 11 glycoproteins with molecular weights ranging from 125 000 to less than 16 000 with two components (40 000 and 33 000) accounting for almost two-thirds of the total stainable material. The FE contained ten glycoproteins that had the same molecular weights as those in the VE. One glycoprotein component underwent a reduction in molecular weight from 77 000 to 67 500 when the VE was converted to the FE. This molecular weight change was interpreted as the probable result of limited proteolysis. In addition, the FE gel electrophoresis patterns contained macromolecular components derived from the cortical granules and jelly layer, J, consistent with the immunodiffusion experiments. These components were absent when the FE was prepared in the absence of Ca2+, suggesting a role for Ca2+ in binding the VE, cortical granules, and J components together. We concluded that the conversion of the glycoproteinaceous VE to FE at fertilization is caused by interaction of the VE with components from the cortical granules and jelly layer J. These interactions are of both a chemical and physical nature.


Stereochemistry of the reaction of sheep liver threonine dehydratase. A nuclear magnetic resonance and optical rotatory dispersion study of its reaction pathway and products. Products, substrates, and inhibitors of the threonine dehydratase from sheep liver (EC 4.2.1.16) have been investigated by proton nuclear magnetic resonance and optical rotation. The alpha-ketobutyrates produced from L-threonine and L-allothreonine in 2H2O have been shown to incorporate a single deuterium into the beta position. The dehydratase forms R-alpha-ketobutyrate-beta-d from L-threonine and L-allothreonine. The alpha protons of the substrates, threonine and allothreonine, do not exchange in the presence of the dehydratase. In the presence of dehydratase, the competitive inhibitors L-cysteine and L-alanine undergo alpha-proton exchange. Highly purified dehydratase has been used to determine kinetic parameters for the usbstrates L-threonine, L-allothreonine, L-serine, and L-chloroalanine. L-Chloroalanine, in addition to being a substrate, inhibits the dehydratase in a manner kinetically identical with that of L-serine.