Actin filaments in sensory hairs of inner ear receptor cells. Receptor cells in the ear are excited through the bending of sensory hairs which project in a bundle from their surface. The individual stereocilia of a bundle contain filaments about 5 nm in diameter. The identity of these filaments has been investigated in the crista ampullaris of the frog and guinea pig by a technique of decoration with subfragment-1 of myosin (S-1). After demembranation with Triton X-100 and incubation with S-1, "arrowhead" formation was observed along the filaments of the stereocilia and their rootlets and also along filaments in the cuticular plate inside the receptor cell. The distance between attached S-1 was 35 nm and arrowheads pointed in towards the cell soma. It is concluded that the filaments of stereocilia are composed of actin.


Experimental autoimmune myasthenia gravis in the rat: a preliminary report. In chronic experimental myasthenia gravis (EAMG) in rat, the decrement of electrical and mechanical responses evoked by maximal repetitive (3-167/sec) stimuli to the nerve was greater in the slow-twitch soleus (SOL) than in the fast-twitch extensor digitorum longus (EDL) muscle. Excitation-contraction coupling was impaired in moderate to severe EAMG, as evidenced by a diminished staircase phenomenon and by diminished posttetanic potentiation (PTP) of twitch tension in two EDLs. The only morphologic abnormality observed was an increase in length of the endplates in the EDLs of those animals that had had an acute phase of EAMG. The latter had more than a 90% reduction in the amplitude of the action potential and in the twitch tension of the EDL when stimuli were applied to the nerve. Stimuli applied directly to the muscle evoked a tetanic tension that was one-third of normal. The staircase and PTP were normal. Necrosis occurred in the endplate and in the adjacent segment of muscle fiber; outside the endplate region, the muscle fiber was normal.


Turnover of rod photoreceptor outer segments. I. Membrane addition and loss in relationship to temperature. Membrane turnover in outer segments of Rana pipiens red rods (ROS) was studied in tadpoles maintained under cyclic lighting (12L:12D) at 23 degrees, 28 degrees, and 33 degrees C. Large fragments (greater than 2 microns in diameter or length) were shed from the ROS tips shortly after the onset of light. These were phagocytized by the pigment epithelium (PE) which caused an increase in the number of phagosomes greater than 2 microns in size (large phagosomes). Large phagosomes were present in highest numbers 2-4 h after light exposure and were degraded by 8-12 h. The proportion of ROS that shed each day after the onset of the light cycle increased with increment increases in temperatures (23 degrees C-18%, 28 degrees C-33%, 33 degrees C-42% per day), resulting, in a reduction in the average interval of time between repeated sheddings (23 degrees C-5.6 days, 28 degrees C-3 days, 33 degrees C-2.4 days) though the average numbers of disks shed from ROS at the various temperatures were not significantly different (23 degrees C-139.5 +/- 5.7, 28 degrees C-129.4 +/- 7.6, 33 degrees C-129.9 +/- 4.8 disks/shed packet). Phagosomes in the PE that were less than 2 microns in diameter (small phagosomes) were present in relatively constant numbers throughout the day, and their numbers increased at higher temperatures. The absence of a concomitant increase in small phagosomes as large phagosomes were degraded indicates that large phagosomes were not the major source of small phagosomes. When the PE was isolated to culture in the absence of the retina, these small phagosomes were degraded. The rate of disk addition to the ROS base was determined by autoradiography after [3H]leucine injection. The number of disks added per day increased with elevations of temperature (23 degrees C-32.4; 28 degrees C-55.9; 33 degrees C-65.5). The average number of disks added to the ROS between repeated sheddings (23 degrees C-181.4; 28 degrees C-167.7; 33 degrees C-157.2) was greater than the number of disks shed after light exposure. Inasmuch as the ROS show no net increase in length during the tadpole stages utilized, the remaining disks must be lost at some other time. Electron microscope analysis revealed the presence of small groups of disks in curled configurations at the tips of ROS, suggesting possible stages of detachment.(ABSTRACT TRUNCATED AT 400 WORDS)


Major histocompatibility complex gene products on macrophages influence T cell activation. Antigen-pulsed macrophages were used to sensitize or elicit sensitivity from mice of different strains to a variety of antigens. The results indicate that sensitization is directed, not to antigen as such, but to a complex structure on the macrophage surface determined partly by the antigen, and partly by a product coded by the major histocompatibility complex. Delayed type hypersensitivity could be provoked by antigen in responder (R) mice and in the F1 between responder and low responder (LR) strains, but not in LR mice unless pretreated by cyclophosphamide. Sensitivity could be transferred to naive LR-strain mice by lymph node cells taken 5 days after sensitization of cyclophosphamide-pretreated LR mice but not of F1 hybrids between LR and R strains. Sensitivity from these could be transferred only to naive F1 or R-strain mice. The results suggest that low responsiveness cannot be accounted for solely in terms of the operation of a cyclophosphamide-sensitive suppressor mechanism. It is postulated that antigen is less immunogenic when presented by LR-strain cells than by R-strain cells.


Cardiac valve replacement in patients over 65 during a 10-year period. From 1967 through 1976, 754 adult patients were subjected to open heart procedures for acquired valvular disease at the University of California, San Francisco, 104 of whom were 66 years of age or over (mean = 70 years). The operative mortality of 15.0% in the elderly group did not differ significantly from that of 14.3% in the entire adult series for the 10-year period. Mortality was consistently higher in combined procedures (multiple valve replacement and valve replacement with coronary grafting). Since the introduction, in 1973, of hypothermic hyperkalemic coronary washout for intraoperative protection of the ischemic myocardium, the hospital mortality rate has decreased to 8.1% overall, 6.0% for isolated aortic valve replacement and 0% for isolated mitral valve replacement in patients over 65. Moreover, the long-term survival following aortic and mitral valve replacement in this series appears to approximate the survival curve of the normal population of the same age. This experience suggests that cardiac surgery has become safer for all patients during the past 10 years and that operative mortality is related primarily to the type and severity of disease rather than to age.


Ventriculography with water-soluble contrast media. In 170 patients 184 ventriculographies with water-soluble contrast media were carried out, 60 examinations with meglumine iocarmate (Dimer-X) and 124 with metrizamide (Amipaque). Grand mal seizures occurred in one patient following the injection of meglumine iocarmate. The indications, technique and diagnostic results are reported.


Ventriculography with metrizamide.

Ventriculography with metrizamide. In 77 patients 85 ventriculographies were performed with metrizamide (Amipaque). No severe complications occurred. The diagnostic accuracy in lesions affecting the midline structures is excellent. However, the rapid passage of the medium from the ventricular system in cases with slight obstructions or small lesions requires a scrupulous technique to enable an exact diagnosis. Due to the difficulty in differentiating between intra- and extra-cerebral location of an expansivity, encephalography should be performed as the first examination whenever possible.


Ventriculography with metrizamide.

Ventriculography with metrizamide. Ventriculography with metrizamide (Amipaque) was performed in 150 patients. Side effects were mild and no severe reactions were observed. The low toxicity and the low epileptogenic activity of this contrast medium make positive contrast ventriculography a safe neuroradiologic procedure.


Alleged inner ear trauma: compensation pending. The English literature on the incidence of nystagmus in normal subjects and in patients with head and/or neck injuries is reviewed. The authors present a series of 202 patients with alleged vestibular trauma. The incidence of spontaneous and positional nystagmus would seem to be no greater in patients with alleged vestibular trauma than that found in the normal subjects, suggesting that these patients do not have objective evidence of vestibular trauma. Objective criteria for diagnosing vestibular abnormalities by electronystagmography are presented. At least 22% of a series of 2408 patients were found to have a functional hearing loss. The importance of looking for functional overlay in patients involved in litigation cases is stressed.


Ventriculography with metrizamide in children. Experiences from 24 ventriculographies with metrizamide (Amipaque) in children are reported. The dose used was 3 to 6 ml 200 mg I/ml in most cases. Ventriculography with metrizamide has been most useful for evaluating obstructive lesions related to the aqueduct or within the third ventricle. The large volume may be required in cases with great ventricular dilatation. Polytomography is a useful addition in many cases but it is not a constant requirement.


Cisternography with metrizamide in cerebellopontine angle tumors. The technique and results of cerebello-pontine angle cisternography with metrizamide (Amipaque) in 34 patients are reported. Fourteen tumors were diagnosed. The internal auditory canal was normal in 19 patients, and in all of them the facial and statoacoustic nerves were visible. One examination was unsuccessful. No serious complications occurred. Headache was recorded in 76 per cent and transient EEG disturbance in 32 per cent of the patients.


EEG disturbances following myelography, cisternography and ventriculography with metrizamide. Two types of EEG abnormalities were recorded after myelography, cisternography and ventriculography with metrizamide (Amipaque) in 292 patients. One, probably without clinical significance, occurred in the EEG of 36 patients as theta waves the following day. The other was of a more serious nature and was recorded as paroxysms of delta waves or spikes in the EEG of 11 patients minutes or some few hours after the myelography. One of these patients developed epileptic seizures.


Clinical experience in North America with metrizamide. Evaluation of 1,850 subarachnoid examinations. The clinical experience of 1 850 diagnostic procedures performed with intrathecal injection of metrizamide (Amipaque) in 1 818 adult patients for various subarachnoid procedures by 21 North American clinical examiners is reported. Adequate radiographic demonstration was achieved in over 90 per cent of all procedures allowing radiologic diagnosis to be made in 99 per cent of the 1 850 examinations. The majority of adverse reactions reported were mild to moderate and transient in nature. No clinically significant changes from baseline were noted in vital signs or neurologic status. The most serious adverse reactions experienced were seizures which occurred in 0.4 per cent of patients. All the patients with adverse reactions recovered without sequelae.


Regional assignment of seven genes on chromosome 1 of man by use of man-Chinese hamster somatic cell hybrids. II. Results obtained after induction of breaks in chromosome 1 by X-irradiation. The position of genes coding for PGD, PPH1, UGPP, GuK1, PGM1, Pep-C, and FH on human chromosome 1 was investigated by analysis of karyotype and enzyme phenotypes in man-Chinese hamster somatic cell hybrids carrying aberrations involving chromosome 1. Suitable hybrid cell lines were obtained by X-irradiation of hybrid cells carrying an intact chromosome 1 and by fusion of human cells from a clonal population carrying a translocation involving chromosome 1 with Chinese hamster cells. The latter human cell population had been isolated following X-irradiation of primary Lesch-Nyhan fibroblasts. In addition, products of de novo chromosome breakage in the investigated hybrid lines were utilized. By integrating the results of these analyses with earlier findings in our laboratory, the following positions of genes are deduced: PGD and PPH1 in 1p36 leads to 1p34; PGM1 in 1p32; UGPP in 1q21 leads to 1q23; GuK1 in 1q31 leads to 1q42; Pep-C in 1q42; and FH in 1qter leads to 1q42.


Mannosidosis: assignment of the lysosomal alpha-mannosidase B gene to chromosome 19 in man. Human alpha-mannosidase activity (alpha-D-mannoside mannohydrolase, EC 3.2.1.24) from tissues and cultured skin fibroblasts was separated by gel electrophoresis into a neutral, cytoplasmic form (alpha-mannosidase A) and two closely related acidic, lysosomal components (alpha-mannosidase B). Human mannosidosis, an inherited glycoprotein storage disorder, has been associated with severe deficiency of both lysosomal alpha-mannosidase B molecular forms. Chromosome assignment of the gene coding for human alpha-mannosidase B (MANB) has been determined in human-mouse and human-Chinese hamster somatic cell hybrids. The human alpha-mannosidase B phenotype showed concordant segregation with the human enzyme glucosephosphate isomerase (GPI) (D-glucose-6-phosphate ketolisomerase, EC 5.3.1.9) but discordant segregation with 30 other enzyme markers representing 20 linkage groups. The glucose-phosphate isomerase gene has been assigned to chromosome 19 in man. This MANB-GPI linkage and confirming chromosome studies demonstrate assignment of the alpha-mannosidase B structural gene to chromosome 19 in man. Since mannosidosis is believed to result from a structural defect in alpha-mannosidase B, these findings suggest that the mannosidosis mutation is located on chromosome 19 in man.


Gene dosage effect for fumarate hydratase (FH; E.C. 4.2.1.2) in partial trisomy 1. A strain of fibroblasts partially trisomic for the larger part of 1q (Norwood and Hoehn, 1974) contains about 1.5 times as much fumarate hydratase (FH) as various control-strains. This gene dosage effect was ascertained by (1) comparative measurements of the specific activity; (2) relating the specific activity of FH to that of reference enzymes, not influenced by the chromosomal anomaly; and (3) by immunoprecipitation methods, using a rabbit antiserum against pig heart FH which cross-reacts with the human enzyme. Among others, this gene dosage effect can be demonstrated numerically by the following parameters: Ratio of the average specific activity of FH in the trisomic strain to that of the control strains: 1.53. Corresponding ratio after dividing FH activity by that of reference enzymes; for acid phosphatase: 1.58, for glutamate dehydrogenase: 1.53. Average ratio of the immunoprecipitation areas obtained upon radial immunodiffusion according to Mancini et al. (1965): 1.56.


Studies of low molecular weight RNA from cells infected with adenovirus 2. I. The sequences at the 3' end of VA-RNA I. VA-RNA I is a low molecular weight RNA produced in large amounts in cells infected with adenoviruses. The 3' terminus of this RNA may represent a transcription termination site. We have demonstrated that this RNA occurs in infected cells in several forms which differ in the number of uridylic acid residues at the 3' ends. The nucleotide sequence of a DNA fragment overlapping the 3' end of VA-RNA I has been determined. The DNA could encode up to 4 uridylic acid residues at the 3' end of the RNA. The DNA sequence shows some similarity to known transcription termination sequences in prokaryotic systems.


Isolation and partial sequence of recombinant plasmids containing human alpha-, beta- and gamma-globin cDNA fragments. Human globin cDNA-derived recombinants with plasmid pCR1 have been prepared for use as specific hybridisation probes and for the partial sequencing of alpha-, beta- and gamma-globin genes.


The nucleotide sequence of the 5' untranslated region of human gamma-globin mRNA. The nucleotide sequence of the entire 5' untranslated region of human gamma-globin mRNA has been determined. This was accomplished by analyzing complementary DNA (cRNA) synthesized from the mRNA with reverse transcriptase. The CDNA was labeled at its 3' end with 32"p using terminal deoxynucleotidyl transferase, digested with the restriction endonuclease Hae III and the end-labeled fragment isolated ans sequenced by the method of Maxam and Gilbert. Including the initiation codon AUG, the 5' untranslated region of human gamma-globin mRNA contains 57 nucleotides, compared to 41 in alpha- and 54 in beta-globin mRNA. There is very little homology between alpha and gamma sequences in the 5' region. There is considerable homology between beta- and gamma-globin mRNAs in the regions proximal and distal to the initiation codon, but the entire sequence shows less homology than the human and rabbit beta-globin mRNAs. The hexanucleotide sequence CUUCUG is found near the 5' ends of all three human globin mRNAs, suggesting a possible role of this sequence or ribosomal binding. Both guanosine and cytidine were found at the 19th nucleotide position from the 5' end of the gamma mRNA. We believe this heterogeneity arises from the difference in nucleotide sequence between the A gamma and G gamma loci.


Sequence of the 3'-noncoding and adjacent coding regions of human gamma-globin mRNA. In cloning human fetal globin cDNA in bacterial plasmids, we obtained a recombinant which contained a fragment of gammg-globin cDNA corresponding to the region from amino acid 99 to the poly A. We determined a sequence of 169 nucleotides which included the complete 3' non-coding region of the gamma-globin mRNA. The codon for amino acid 136 was GCA, indicating that this cloned fragment was derived from the Agamma-globin gene. In conjunction with the surrounding sequences, the GCA codon provides the Agamma-species with a unique CTGCAG hexanucleotide that is recognized by the restriction enzyme Pst I. The 3'-untranslated region of the gamma-globin mRNA consists of 90 nucleotides, and shares little homology with that of the human beta-globin mRNA. As in other mammalian mRNAs, a symmetrical sequence and the hexanucleotide AAUAAA are present.


Failure of urethane anesthetic to block induction of pineal serotonin N-acetyltransferase activity in the rat. Ability of urethane anesthetic to block induction of pineal serotonin N-acetyltransferase (SNAT; E.C.2.3.1.5) activity was measured in individual rat pineal glands in animals receiving urethane (25% w/v, IP, 1.2 g/kg) or saline, 6 hr prior to sacrifice. Using a radioenzymatic assay, SNAT determinations were made twice daily (at 1200 or 2400 hr) immediately after the sacrifice of each animal. The results show that urethane had no effect on the induction of SNAT activity: (1) implying that the neural activity of those structures involved in induction of SNAT activity (e.g., suprachiasmatic nucleus) is not substantially altered by this anesthetic and (2) suggesting that the central blockade of ovulation by urethane does not include alterations in suprachiasmatic nucleus activity.


Neuron activity in rat hippocampus and motor cortex during discrimination reversal. Chronic unit activity and gross movement were recorded from rats during two discrimination reversals in a classical appetitive conditioning situation. The anticipatory movement decreased in response to the former CS+ tone and increased to the previous CS- tone after each reversal. Hippocampus and motor cortex were differently related to these two kinds of behavioral change. Response rates of hippocampal neurons were more closely related to the increased movement response to the former CS- which now signaled food. Motor cortex neuron responses were more closely correlated with the decrease in movement responses to the former CS+ which became neutral after the reversal. It appeared that hippocampal neurons could have been involved in one cognitive aspect of the situation, motor cortex neurons in another. The data were related to current functional concepts of these brain regions.


Fornix afferents to the anteroventral thalamic nucleus: an EM study in the rat. Three types of synapses occur in the anteroventral thalamic nucleus (AVN). Type 1 consists of small (0.5-0.8 microns) axonal endings densely packed with spherical synaptic vesicles. They form markedly asymmetrical synaptic contacts with distal portions of dendrites. Degenerative changes in these axons following destruction of the fornix identify them as the endings of the subicular projection to AVN. Type 2 synapses consist of large (1.0-1.5 microns) axonal processes containing spherical vesicles which form asymmetrical synapses on more proximal dendrites. Type 3 endings consist of large unidentified processes containing spherical, and occasionally flattened, synaptic vesicles forming symmetrical contacts with the largest stem dendrites. Neither of these synaptic types were modified by fornix lesions. The synaptic arrangements within AVN are simpler than other thalamic nuclei in that serial synapses and synaptic glomeruli are not present.


d-Amphetamine and ethanol: a drug-drug interaction study. The in vivo and in vitro effects of d-amphetamine on the specific activities of endogenous liver alcohol-(L-ADH) and aldehyde dehydrogenase (L-ADH) were studied using rat and mouse liver enzyme preparations. d-Amphetamine, 10(-4)M, noncompetitively inhibited rat and mouse L-ALDH in vitro. Mitochondrial L-ALDH was not affected by d-amphetamine in vivo and in vitro in both species studied. A noncompetitive inhibition of mouse L-ADH, occurred in vivo 16 hr after administration of d-amphetamine, 4 mg/kg, IP. A moderate, but not significant decline in specific activity of rat L-ADH was determined after injection of d-amphetamine, 4 mg/kg IP, showed a significant decrease in their blood and brain content of exogenously administered ethanol from corresponding saline-treated controls for the 60 min and 90 min periods after ethanol administration, respectively. The results show species differences in d-amphetamine action on liver enzymes studied and indicate that d-amphetamine can interfere in the metabolism of ethanol.


14C-dopamine microinjected into the brain-stem of the rat: dispersion kinetics, site content and functional dose. A morphological analysis was undertaken of both the dispersion characteristics and tissue content of dopamine (DA) microinjected acutely into the brain-stem of the anesthetized rat. 14C-DA, with a specific activity of 56-62 mCi/mMol, was infused unilaterally into the pars compacta of the substantia nigra in one of four test volumes: 0.5, 1.0, 4.0 or 8.0 microliters. The concentration of the 14C-DA solution was 1.0 microCi/microliter, equivalent to 3.01 micrograms/microliters, which was delivered at an injection rate of 1.0 microliter per 45 sec. At an interval of either one min or 15 min following the microinjection, the rat's brain was removed rapidly from its calvarium, flash frozen and then cut in the coronal plane on a freezing microtome in 500 micron slabs. After each of the respective serial slabs was mounted on glass, the Eik Nes-Brizzee trochar technique for the discrete removal of tissue samples was used to obtain 0.5 mm dia. cylindrical plugs of meso-diencephalic tissue at distances from the site of injection ranging from 0.5 to 2.5 mm, center to center. Each sample plug was subsequently solubilized and 14C-DA activity quantitated by liquid scintillation spectrometry. The results show that regardless of volume, the spatial patterning of the microinjected solution assumes a tear-drop or pear shape, not a sphere. Further, as the volume of the injection is increased from 0.5 to 8.0 microliters, the magnitude of the dispersion of 14C-DA is enhanced throughout the surrounding parenchyma, but not in a linear fashion.(ABSTRACT TRUNCATED AT 250 WORDS)


Oestrogenic influences on the electrical activity of the olfactory pathway. The influence of the oestrogenic hormones over the spontaneous and induced activity of the olfactory pathway was studied in normal female cats. Electrodes were placed chronically or acutely in the olfactory bulb (OB), olfactory tubercle (OT) and in the prepyriform cortex (PPC). Oestrogenic hormones were applied locally in the posterior hypothalamic region. Recordings were made during the two different phases of the oestral cycle. In addition, another group of castrated animals was studied. The oestral phase was induced in these cats by the subcutaneous administration of 17-beta-oestradiol. Results indicate that the pattern of the electroencephalographic spontaneous activity as well as the response induced by hypothalamic stimulation changed. The number of the bursts for each 10 sec trace was higher in oestrus than in anoestrus for all the structures studied. The duration of each burst also changed, being shorter in oestrus than in anoestrus cats. The threshold for significant bursting in the olfactory structures following hypothalamic stimulation was lower in oestrus than in anoestrus. The evoked potentials recorded in the same three olfactory structures by hypothalamic stimulation exhibited changes in correlation with the hormonal administration. In all the structures studied the amplitude of the different components of the evoked potentials increased immediately after the hormones were administered. However, the most dramatic increase was observed in the olfactory tubercle. In order to further investigate these changes in acute preparations, a study evaluating the excitability changes was conducted. Applied pulse pairs, with different interpulse intervals between 200 to 1000 msec, were delivered in the hypothalamus before and after 200 micrograms of the hormone were administered into the posterior hypothalamus.(ABSTRACT TRUNCATED AT 250 WORDS)


Dopamine release within forebrain sites perfused with tetrahydroisoquinolines or tryptoline in the rat. The localized effect of amine-aldehyde condensation products on the endogenous release of dopamine (DA) was examined in dopaminergic-rich areas of the brain. In the unanesthetized rat, dopaminergic stores were labeled by 14C-dopamine (1.0 microCi) microinjected in a volume of 1.0 microliters into the caudate nucleus, tuberculum olfactorium, nucleus accumbens or other contiguous sites within the ventral forebrain. Subsequently, each labeled site was locally perfused with an artificial CSF, by means of push-pull cannulae, at a rate of 25.0 microliters per min. Two tetrahydroisoquinolines, tetrahydropapaveroline and 4,6,7-TH-tetrahydroisoquinoline, as well as the tetrahydro-beta-carboline, tryptoline, significantly enhanced the release of 14C-DA when each alkaloid metabolite was perfused in a concentration of 250 nanograms per min. Tetrahydroisoquinoline was without effect on the local kinetics of 14C-DA, whereas salsolinol had an intermediary action on DA release. These findings suggest that an alkaloid conjugate when given in the cerebral ventricle could exert its central effects by way of a dopaminergic mechanism in the telencephalon.


Measurement of intracranial pressure in the unanesthetized rabbit. A method is described for the measurement of intracranial pressure in the unanesthetized, minimally restrained rabbit utilizing a modified subarachnoid screw system. The pressures are transmitted from within the cranium via a flexible saline filled catheter to a fixed external pressure transducer. An index of the relative vertical position of the animal's skull as compared to the fixed transducer is given by means of a second open-ended pressure measuring catheter, the open end of which is fixed to the subarachnoid screw assembly on the animal's skull. The system was found to be a reliable method of measurement of intracranial pressures in the minimally restrained rabbit, and could easily be adapted to other animal species. The method is currently being used to assess the effects of fever on intracranial pressures.


Analysis of peptides from pigeon cerebrum: a comparison of sacrifice methods. Peptides were extracted and fractionated from pigeon cerebrum and analyzed by thin-layer chromatography, polyacrylamide disc gel electrophoresis, and amino acid analysis. A highly consistent pattern of peptides in the 700-5000 molecular weight range was obtained from each cerebral sample. A comparison was also made between cerebral peptide patterns obtained from animals sacrificed by decapitation with those from animals sacrificed by microwave irradiation. Amino acid analysis of the peptide fraction from microwaved and decapitated animals revealed slight differences in the amount and incidence of various amino acids. These differences may be due to post-mortem proteolysis in the cerebra of decapitated pigeons prior to homogenization and freezing.


Analgesia produced by direct injection of morphine into the mesencephalic reticular formation. Morphine administered intracerebrally (IC) (10 micrograms as the base on each side) into the MRF produced a significant dose dependent elevation of nociceptive threshold (i.e., analgesia) on the tail flick test and hemostat pinch test. However, morphine IC at lower doses had no analgesic effect. After morphine was injected IC (10 micrograms, bilaterally) into the MRF, naloxone, a specific narcotic antagonist, administered either IC at the same site (15 micrograms, bilaterally) or subcutaneously (10 mg/kg), antagonized the antinociceptive effects of morphine. Thirty percent of the animals given bilateral microinjection of 10 micrograms of morphine displayed hyperreactivity to mild stimuli. This hyperreactivity was not attenuated by large IC or systemic doses of naloxone. It was concluded that the MRF is a site where morphine may act to produce analgesia by a specific narcotic mechanism of action.


Reduction of distress vocalization in chicks by opiate-like peptides. All the opiate-like peptides we tested (Met-enkephalin, (D-Ala2)-Met-enkephalin-NH2, beta-endorphin, (D-Ala2)-beta-endorphin, (D-Ala2)-alpha-endorphin, (D-Ala2)-gamma-endorphin) were capable of reducing distress vocalizations (DV's) in socially-isolated chicks when injected into the vicinity of the fourth ventricle in doses as low as 100 picomoles. All of these substances were at least as potent as equimolar doses of morphine sulfate. In general, DV's were a more sensitive measure of opiate-like peptide effects than reductions in body temperature. In a more limited study using peripheral injections, it was determined that (D-Ala2)-Met-enkephalin at doses of 400 nanomoles/kg, like morphine sulfate, was more effective in reducing DV's, than an equimolar dose of beta-endorphin. beta-endorphin was not as effective via a peripheral route as it was via central administration.


Procedure for the decerebration of the rat. A procedure for the decerebration of the rat was devised. The internal and external carotid arteries were ligated under halothane anesthesia. A transection was made at the mid-collicular level of the brain and the transected forebrain was removed. The posterior communicating arteries were occluded by suction and oxidized cellulose was packed around these vessels. The cut surface of the brain was covered with oxidized cellulose and the cranial cavity was filled with cotton balls. Head-up tilt, carotid occlusion and sodium cyanide responses indicated that the orthostatic, baro-, and chemo- reflexes were intact in these preparations. The stability of the preparation (for at least 8-10 hr) and the integrity of these reflexes indicate that the decerebrate rat may be a suitable alternative for studies involving brain stem regulatory mechanisms in this species when it is desirable to avoid general anesthetics.


Caudate afferents from the retrorubral nucleus and other midbrain areas in the cat. In order to clarify which midbrain areas project to the caudate nucleus (Cd), horseradish peroxidase (HRP) was deposited in the Cd of 13 cats. Midbrain sections were reacted with diaminobenzidine and sometimes with cobaltous chloride to intensify the reaction, and then examined for retrograde labelling. Afferents to the Cd were found arising from the following ipsilateral midbrain areas: pars compacta, pars reticulata, and pars lateralis of the substantia nigra (SN), ventral tegmental area of Tsai, interpeduncular nucleus, central tegmental field, and the retrorubral nucleus (RR) of Berman, which lies caudal to the red nucleus and dorsal to the medial lemniscus. HRP labelled cells were also found within the "cell bridges" which lie between SN and RR, composed of neurons among the fascicles of the medial lemniscus. Acute lesions placed in the area of the lateral hypothalamus and medial forebrain bundle prior to HRP injection blocked HRP uptake in the ipsilateral SN, RR, and cell bridges. Thus, the SN, RR and connecting cell bridges share, at least in part, a pathway to the Cd in the cat.


Radioimmunoassay of DSIP-like material in rat brain. Antibodies generated in a rabbit by immunization with synthetic delta-sleep inducing peptide (DSIP) showed no cross-reactivity with 19 naturally-occurring peptides or analogues and were used to establish a radioimmunoassay. Since DSIP is not readily iodinated by conventional methods, N-Tyr-DSIP was synthesized to prepare the tracer; the dose-response curve for N-Tyr-DSIP was exactly parallel to that for DSIP. With this assay, DSIP-like immunoreactivity was detected in the brain of rats. The highest brain values were found in the thalamus (11.9 +/- 1.3 pg/mg). The widespread presence of DSIP-like material throughout the body as well as the shared amino acid sequences of DSIP with other proteins suggest caution in defining the material being measured by this new assay. Nevertheless, the demonstration of DSIP-like activity in brain tissue and elsewhere warrants consideration of functions in addition to a possible role in sleep.


Micro-injections in the avian embryo: technical notes. Procedures and instrumentation are described for making micro-injections into early avian embryos. Tools for implementing injection include support probes and micro-cannula adapters, both of which greatly reduce embryonic insult. Using these instruments, viability following certain early embryonic injections has been shown to increase from 29 to 95%.


Utilization of 14C-tyrosine in brain and peripheral tissues of developmentally protein malnourished rats. Prior studies of developmentally protein malnourished rats have reported substantial changes in brain and peripheral utilization of 14C-leucine, 14C-phenylalanine, and 14C-tryptophan. In the present study rats born to dams fed a low protein diet (8% casein) compared to the offspring of control rats fed a normal diet (25% casein) showed few significant differences in the uptake and incorporation of 14C-tyrosine into brain and peripheral tissues from birth to age 21 days. At birth, the 8% casein pups exhibited significant decreases in brain and peripheral tissue incorporation of tracer only at short post-injection times (10 and 20 min), but not at longer intervals (90 and 180 min). During ontogenetic development (Days 5-21), the 8% casein rats showed significant increases in uptake of 14C-tyrosine into the brain and peripheral tissues on Day 11 and a significantly higher percent incorporation of tracer into brain protein on Day 21 as compared to the 25% casein rats. For the most part, there were no significant changes in incorporation of radioactivity in peripheral tissues for the 2 diet groups on these post-birth days. Overall, the data indicates that developmental protein malnutrition causes relatively fewer changes in brain and peripheral utilization of the semi-essential amino acid tyrosine than those observed in previous studies with essential amino acids.


Distribution of prostaglandins E and F in different regions of the rat brain. The regional distribution of prostaglandins E and F in 24 different areas of the rat brain was studied. For bioassay the rat uterine horn preparation was used. Prior to bioassay the samples were purified with organic solvents and in some cases by chromatography on silica as well. For measurement of PGF2 alpha content in certain areas a radioimmunoassay was used. Each region was found to contain prostaglandins but in uneven concentrations. Highest concentration, related to the protein content, was present in the median eminence, 57.6 ng PGE and 34.4 ng PGF per mg protein. Parietal cortex, olfactory tubercle, cerebellar cortex, caudate nucleus, substantia nigra, and paraventricular nucleus are also rich in prostaglandins. Low values were found in frontal cortex, thalamus, and septum. The distribution of prostaglandins throughout the areas examined do not show any direct correlation with that of transmitter substances. The uneven distribution can be attributed to differences in the endogenous biosynthetic capacity of each region.


Suprachiasmatic nucleus responsiveness to photic and basal hypothalamic stimulation. In view of the demonstrated role of light and of the suprachiasmatic nucleus (SCN) in the maintenance of circadian rhythms related to endocrine functions and as the mediobasal hypothalamus (MBH) controls neuroendocrine activity, the effects of light and MBH stimulation on the electrical activity of SCN neurons were studied in rats. Out of 253 cells studied in the SCN, 32 cells were antidromically activated, while 65 cells responded orthodromically to MBH stimulation. In another series of experiments out of 95 suprachiasmatic neurons, 11 were antidromically activated by MBH stimulation and 18 different SCN cells responded to continuous light. The present data, which demonstrate photic responses in suprachiasmatic neurons and direct projections from the SCN to MBH, are discussed in view of the current knowledge on the role of the retinohypothalamic tract and the SCN in the control of circadian rhythms.


Three-dimensional plotting of neuronal population response patterns. A computer technique is described for generating three-dimensional plots of the distribution of spike activity in neuronal populations. Data obtained from single neurons in the sensorimotor cerebral cortex of cats are used to plot spike density with respect to depth in the cortex and time following a cutaneous stimulus. The computer programs include a masking (hidden line) subroutine and produce a cross-hatched plot showing the rise and fall of neuronal activity in both space and time. By manipulation of the data matrix and by axis rotations, the plot can be viewed from any desired perspective. This method constitutes a powerful technique for analyzing the spatio-temporal response patterns of neuronal populations. The significance of neuronal population responses for encoding stimulus information and predicting behavior is discussed. Note: Details concerning the computer programs developed for this plotting technique may be obtained from the Division of Systems and Computer Services, Medical College of Georgia, Augusta, GA 30901.


Development of habituation in the dentate gyrus of rat: physiology and anatomy. The physiological development of monosynaptic response habituation in the rat dentate gyrus was compared to morphological development. Rapid Golgi techniques were coupled with in vitro studies of dentate granule cell habituation to several frequencies and intensities of monosynaptic excitation. Except for the youngest group, the degree of habituation increased as a function of age, paralleling the morphological development.


Effect of avian basal forebrain lesions, including septum, on heart rate conditioning. The possible involvement of basal telencephalic structures in visually conditioned heart rate change (established by pairing light and foot-shock) was studied in 156 pigeons by evaluating conditioning performance following lesions of the septum or lobus parolfactorius. Extensive destruction of the septal complex had no effect on either the orienting response or the development of the conditioned response. Lesions of the lobus parolfactorius did not affect the orienting response or overall conditioned response levels, but it did slightly prolong the latency of the conditioned heart rate change. It is concluded that the septum, despite its being cardioactive, is not involved in conditioned heart rate change and that the lobus parolfactorius participates minimally. Thus, of the principal limbic structures of the avian telencephalon, only the amygdalar homologue appears critical in this defensive conditioning task [3].


Principal neurons of the basilar pons as the source of a recurrent collateral system. The basilar pontine nuclei in the opossum are composed of two general categories of neurons, intrinsic cells and the principal or projection neurons. Observations from Golgi material indicate that principal neurons whose primary axons project to the cerebellar cortex may also give rise to recurrent branches distributing within the pontine gray. Such collaterals were observed to arise near the soma and at some distance from the cell body of the parent axon. The electron microscopic correlate of such a system was identified in the basilar pontine neuropil in animals subjected to lesions of the cerebellar cortex. These lesions destroyed mossy terminals and their parent axons and thus initiated a retrograde reaction in basilar pontine projection neurons which manifested itself in the form of morphologic alterations observed in somata, dendrites, and a class of axonal boutons. Similar altered axon terminals were not observed in control material and did not correspond to the terminals of cerebello-pontine axons described in previous work. It was therefore suggested that such boutons represented the terminals of the recurrent collateral system observed in Golgi material.


Anatomic evidence suggestive of dendrodendritic synapses in the opossum basilar pons. In Golgi-prepared material, opossum basilar pontine intrinsic neurons measured less than 22 microns and gave origin to only two or three primary dendrites which gradually tapered in diameter, branched infrequently and exhibited few spines or protrusions. Characteristically, such neurons gave rise to several axon-like processes which might take origin from proximal or distal dendrites as well as the soma. Morphologically similar neurons have been observed in several other regions throughout the CNS where they have been shown to be a source of presynaptic dendritic elements. That opossum basilar pontine intrinsic neurons might also give rise to presynaptic dendrites was supported by the following electron microscopic observations. Profiles containing a small cluster of pleomorphic vesicles, occasional ribosomes and numerous microtubules formed synaptic active sites in which the membrane densities were intermediate between Gray's type I and type II. Such presumed presynaptic dendritic profiles were observed to participate in serial synaptic arrangements in which they were always postsynaptic to a round vesicle bouton and presynaptic to another dendritic element. The above features are compatible with electron microscopic descriptions of presynaptic dendrites in other CNS regions and thus suggest that basilar pontine intrinsic neurons may represent a source of presynaptic dendritic elements.


Cocaine and pseudococaine: comparative effects on electrical after-discharge in the limbic system of cats. The effects of cocaine and its dextroisomer pseudococaine on electrical after-discharge (AD) evoked by electrical stimulation of the hippocampus or amygdala were studied in cats with electrodes implanted in the brain. Intravenous injection of cocaine (2.0 to 4.0 mg/kg doses) produced a suppressive effect on the AD while producing low-voltage fast waves (LVFWs) in the electrical activities of the brain (EEG) associated with behavioral excitation. In contrast, pseudococaine at the same dose as cocaine failed to show a significant suppressive effect on the AD except at high doses (5.0 mg/kg). Pseudococaine produced high-voltage slow waves (HVSWs) in the EEG associated with behavioral depression. A linear dose-response relationship was observed for the suppressive effect of cocaine on the AD. The results suggested that the limbic system may be involved as a primary site of action of cocaine in the central nervous system (CNS).


Intracranial self-stimulation site specificity: the myth of current spread. Locus-specificity of intracranial self-stimulation (ICSS) was determined for 78 electrode placements using monophasic stimulation. ICSS rate comparisons between each pole of a bipolar electrode when each served as cathode were made when the anodal source was either the other pole of the bipolar electrode or a skull screw. In hypothalamic areas, medial forebrain bundle electrode tips elicited significantly higher rates than electrode tips of the same bipolar electrodes located in perifornical or far-lateral diencephalic placements. In turn, perifornical electrode tips elicited higher rates than more dorsally or medially placed tips. In dorsal pontine areas, locus coeruleus electrode tips elicited significantly higher rates than more medially or laterally placed tips. In periaqueductal midbrain and substantia nigra placements, tips located along the midline or in the substantia nigra elicited significantly higher rates than tips located lateral to or ventral to those respective structures. Anodal locus did not change these results. These results suggest that ICSS behavior is delimited by and corresponds to neuroanatomically discrete entities and that cathodal, rather than anodal factors seem to most crucially determine ICSS integrity.


Adrenergic stimulation of the paraventricular nucleus and its effects on ingestive behavior as a function of drug dose and time of injection in the light-dark cycle. The paraventricular nucleus (PVN) has been identified as the most responsive site in the brain for eliciting feeding and preprandial drinking responses with local injection of norepinephrine (NE) in satiated rats. The present study examined these phenomena as a function of dose and time of injection in the light-dark cycle. The lowest effective doses for producing these ingestive responses with exogenous NE in the PVN are found to be between 5.6 and 16.9 ng for preprandial drinking and between 1.0 and 4.2 ng for feeding. Tests with NE injection into the PVN at different times in the light-dark cycle indicate that an increase in feeding effect can occur in the dark as well as in the light and at varying levels of food intake baseline and with solid and liquid food. Site of injection in the brain is believed to be a crucial factor in determining the nature of NE's effects on feeding behavior.


DNA synthetic ability in cerebella from temperature and handling stressed neonatal rats. DNA synthetic ability of neonatal rat cerebellum was studied in two stressful conditions (cooling and excessive handling) which may be inadvertently imposed during the course of certain experiments. A rate study of the ability of animals handled excessively for 10 min to incorporate 14C-thymidine into the DNA tissue fraction (DNAF) from the acid soluble tissue fraction (ASF) of the cerebellum revealed a decreased DNA synthetic ability which lasted for 1 hr. Animals with very low core temperatures incorporated little 14C into the DNAF from the ASF 2 hr after an IP injection of 14C-thymidine. There was a proportional decrease in the ability to synthesize DNA as core temperature decreased.


Quantification of cell death in developing cerebellum by a 14C tracer method. To study the question of whether or not cell death contributes significantly to normal or stressed postnatal brain development in a way which is biochemically quantifiable, we carried out an experiment to assess the amount of cell death in developing cerebellum. By measuring the loss of DNA content and the loss of 14C from labelled thymidine previously incorporated into the DNA fraction (DNAF) in X-irradiated neonatal animals, shown by histological methods to have cell death to the degree of degranulating the external granular layer (EGL), we showed that when cells die both label and DNA content are greatly decreased in the cerebellum. Experiments on both normal and malnourished animals showed that cell death does not contribute significantly to cerebellar development in either malnutrition-stressed or normal animals. Here, we present a biochemical tool for assessing cell death and evidence that cell death does not contribute significantly to cerebellar development.


The topographic organization of hypothalamic and brain stem projections to the hippocampus. Direct projections primarily ipsilateral to hippocampus from medial septal, diagonal band, supramammillary, submammillothalamic, locus coeruleus, and dorsal and medianus raphe nuclei were demonstrated. The locus coeruleus projects primarily through the cingulum and fornix superior to the dorsal posterior hippocampus, with its terminal fields in the stratum lacunosum moleculare of the subiculum and areas CA 1-CA 2 of the dorsal posterior hippocampus. LC projections to the granular layer of the dentate hilus were not found. Raphe nuclei project through the cingulum, fornix superior, and primarily the fimbria, to the dorsal and ventral posterior hippocampus, with their terminal fields in the stratum lacunosum moleculare of the dorsal posterior subicular region, stratum radiatum of CA 1-CA 3 in the dorsal hippocampus, and the stratum polymorph of the dentate gyrus, primarily in its superficial part. Raphe projections to the anterior hippocampal rudiment were found. However, no projection was found to the subiculum of the ventral posterior hippocampus, nor to stratum oriens. Hypothalamic nuclei project through the fornix superior and the fimbria, mainly to the dorsal posterior hippocampus with abundant terminal fibers in the depth of the dentate hilus. Smaller cells in these hypothalamic nuclei appear projecting to the ventral hippocampus. The number of neurons in the entorhinal area, the diagonal band, and the hypothalamic nuclei projecting to the hippocampus suggests these groups as the main sources of the extrinsic hippocampal afferents. In addition, they may also serve as relay stations for inputs from more caudal nuclei, and the topographic organization of their terminal fields as described herein may have important functional implications.


Covalent coupling of rat liver phenylalanine hydroxylase. Optimal conditions have been determined for the coupling of rat liver phenylalanine hydroxylase (Phe H) to cyanogen bromide-Sepharose 4B. When 8 mg of ligand was reacted with 100 mg of matrix, 20 to 30 percent of the initial enzyme activity was covalently bound along with 90 percent of the protein. The coupled enzyme showed greater thermal stability from 40 degrees to 60 degrees, a broader base of optimal pH activity from 5.8 to 10.0, more resistance to proteolysis and less inhibition by various inhibitors. The uncoupled enzyme exhibited greater storage stability at 25 degrees after 24 hr and at 0 degrees after 18 days. Alteration of the microenvironment by introduction of sulfhydryl groups or of carriers having positive or negative charges had variable effects on the hydroxylase activity.


Unilateral cortical spreading depression in the rat: effects on feeding, drinking and other behaviors. The effects of unilateral cortical spreading depression (CSD) on feeding, drinking and other behaviors were studied in rats. CSD waves were elicited by a microinjection of KC1 solution through an implanted cannula. The animal's behavior and slow potential changes (SPC) accompanying CSD were then simultaneously monitored for 20 min. Intensive face grooming was observed while the CSD wave-front propagated across the sensorimotor cortex. Animals commenced their first feeding or drinking shortly after the invasion of the negative SPC into the frontal cortex of Krieg's area 6 and 10 [14] had reached its maximum height. A two-way switching effect between the first feeding or drinking and quiescence (freezing) and a close relationship between their onsets were observed. The results are discussed specifically in relation to cortico-hypothalamic pathways.


Cefoxitin for treatment of infections due to anaerobic bacteria. Twenty-seven patients with 29 infections due to anaerobic or mixed anaerobic-aerobic bacteria were treated with cefoxitin. From the 27 patients 99 isolates of anaerobic bacteria and 70 isolates of aerobic bacteria were recovered. Twelve pleuropulmonary infections, 12 soft tissue infections, three bone infections, and two intraabdominal infections were treated. Twenty-six of the 29 infections were completely or partially eradicated; the remaining three infections were unchanged. One-third of the patients experienced phlebitis; in two instances severe phlebitis caused discontinuation of therapy. In this study cefoxitin appeared to be an effective single agent for the therapy of infections due to specific anaerobic bacteria or to a mixed flora of anaerobic and aerobic bacteria.


Penetration of cephalothin and cefoxitin into experimental infections with Bacteroides fragilis. The in vitro activities of cephalothin and cefoxitin against Bacteroides fragilis were studied by time-kill curves and measurement of residual drugs in culture supernatants. Cefoxitin was bactericidal, causing a decrease of 10(7) in viable counts over 24 hr. Cephalothin caused an initial decrease of 10(2) B. fragilis at 2 hr; this change was followed by growth of the organism within 24 hr back to the number present before addition of cephalothin. The concentration of cephalothin in broth decreased rapidly within 2 hr and was undetectable within 24 hr, whereas the level of cefoxitin decreased only 25% over the 24-hr period. Penetration of these drugs into perforated ping pong balls implanted intraperitoneally in rabbits was studied. Three weeks after implantation the reservoirs were infected with B. fragilis. After intramuscular administration of five doses of antibiotic, the penetration of cephalothin, as measured by bioassay, in uninfected and infected capsules was 16% and 2%, respectively, of the peak serum concentration; similar findings were noted with cefoxitin. For determination of the rate of breakdown within the infected site, radiolabeled antibiotic was injected into the capsule, and the concentrations of bioactive and radioactive drug were determined. With radiolabeled cephalothin there was a rapid decrease in bioactivity during the initial 60 min, and no active drug was measurable after 2 hr. In contrast, only 40% of cefoxitin was inactivated at the end of 6 hr. The results indicate that levels of cephalothin and cefoxitin are reduced significantly in sites infected with B. fragilis. The decrease appears to be mediated by both a decrease in penetration and inactivation at the site of infection.


Treatment of significant bacterial infections in adults with cefoxitin. Cefoxitin is a new antimicrobial agent derived from cephamycin C. Fifty-four hospitalized patients with 63 clinically significant infections were treated with cefoxitin. Fifty-four infections (86%) were cured by therapy with cefoxitin alone or together with local therapy (i.e., heat, elevation, or surgical drainage). Six infections (10%) were not eradicated by the therapy used. Three patients could not be evaluated. Addition of oral probenecid therapy produced increases in levels of cefoxitin in serum and increased the inhibitory and bactericidal levels of cefoxitin in serum to more favorable ratios. Adverse side effects were seen in 19 patients and included positive direct Coombs' tests, increased eosinophil counts, skin rashes, vasculitis, phlebitis, elevation of liver enzymes, and drug-induced fever.


Pharmacokinetics of cefoxitin in normal subjects and in patients with renal insufficiency. The pharmacokinetics of a single intravenous dose of 30 mg of cefoxitin/kg given to normal subjects and to patients with various degrees of chronic renal insufficiency were studied. The serum half-life of cefoxitin was correlated inversely with the rate of creatinine clearance, and the half-life increased exponentially when the creatinine clearance rate fell below 30 ml/min per 1.73 m2. The elimination of cefoxitin into urine was rapid and extensive in patients whose creatinine clearance was greater than 30 ml/min per 1.73 m2. In patients with more severe degrees of renal impairment, renal clearance of drug was diminished markedly (less than 20% of the administered dose was excreted in 4 hr as compared with greater than 60% in the former group). The clearance of cefoxitin can be increased fivefold by hemodialysis in patients with end-stage renal disease.


Penetration of cefoxitin into cerebrospinal fluid of dogs with and without experimental meningitis. The penetration of cefoxitin into the cerebrospinal fluid (CSF) after slow intravenous infusion of 50-100 mg/kg over 1 hr was studied in normal dogs and in dogs with meningitis experimentally induced by intrathecal injection of 10(9) Staphylococcus aureus. With healthy dogs the peak CSF concentration of 1 microgram of cefoxitin/ml was found to correspond with a serum level of 120 micrograms/ml. With administration of probenecid the CSF level was 1.5 micrograms of cefoxitin/ml. Doubling the dose from 50 to 100 mg/kg resulted in a CSF concentration of 2 micrograms of cefoxitin/ml. In the CSF of animals with meningitis, a peak concentration of 10 micrograms of cefoxitin/ml was obtained 90 min after the start of the infusion, and 5 micrograms/ml was still present at 240 min. The peak level in CSF of animals with meningitis was about 10% of the simultaneous level in serum.


Therapy of skin, soft tissue, and bone infections with cefoxitin sodium. Twenty-seven patients with skin and soft tissue infections, including three with contiguous osteomyelitis, were given cefoxitin intravenously or intramuscularly; the infections of 25 (93%) were resolved with cefoxitin therapy. Etiologic agents included staphylococci, streptococci, Enterobacteriaceae, and anaerobes. Susceptible pathogens were inhibited by less than or equal to 8 micrograms of cefoxitin/ml. This level of drug was surpassed by mean peak serum concentrations eight- to 12-fold after intravenous infusions and two- to threefold after intramuscular injections and resulted in eradication of susceptible organisms from lesions during treatment. Intravenously administered cefoxitin was well tolerated, although eosinophilia, phlebitis, elevation of levels of hepatic enzymes, and a positive direct Coombs' test were observed. Intramuscular injections of cefoxitin in 0.5% lidocaine caused pain and induration and thus were poorly tolerated.


Penetration of cefoxitin into human cerebrospinal fluid: comparison with cefamandole, ampicillin, and penicillin. Single, intravenously administered doses of cefoxitin and cefamandole did not penetrate into cerebrospinal fluid of normal humans. Multiple-dose administration with or without probenecid facilitated penetration of both antibiotics into the cerebrospinal fluid. Preliminary data showed that cefoxitin penetrated into cerebrospinal fluid of patients with inflamed meninges even when administered in a single dose without probenecid. However, the concentrations of cefoxitin in the cerebrospinal fluid of the individuals studied were not within the therapeutic range.


Cefoxitin therapy for bacterial endocarditis. Of 22 patients who were suspected of having bacterial endocarditis and who were treated with cefoxitin intravenously (8-12 g per day), 12 were evaluated for responses to therapy. Ten patients had infections due to a single pathogen, and two had polymicrobial infections. Staphylococci were isolated from eight patients, and streptococci from four; both of these pathogens were susceptible to 2-16 micrograms of cefoxitin/ml. Staphylococcus aureus and four strains of anaerobic bacteria, including Bacteroides fragilis (minimal inhibitory concentration, 32 micrograms/ml), were isolated from one patient. The average level of cefoxitin in serum was 32.8 micrograms/ml (range, 14.5-64 micrograms/ml) at 1 hr after an intravenous dose of 2 g; after 5 hr the average level in serum was 8.5 micrograms/ml (range, 2-20 micrograms/ml). The mean (+/- SD) level of cefoxitin in myocardial tissues from eight rabbits at 1 hr following a 250-mg/kg dose of the antibiotic was 4 +/- 0.5 micrograms/g. On the average, patients were treated for 29 days (range, 14-40 days), and they became afebrile in 6.2 days (range, three to 20 days). Both clinical and microbiologic responses to cefoxitin therapy were excellent in 10 patients with monobacterial infections. Both patients with polymicrobial infections were not cured. One, who was infected with a mixed flora of anaerobes, died; the other was cured after surgical valvectomy. These results suggest that cefoxitin is effective in the treatment of endocarditis due to a single susceptible organism but that this antibiotic should be used with caution in patients whose endocarditis is caused by a mixed population of bacterial pathogens.


Treatment of septicemic patients with cefoxitin: pharmacokinetics in renal insufficiency. Experience with cefoxitin in the treatment of septicemia of diverse etiologies in 11 patients is reviewed. Results of a small study of the pharmacokinetics of cefoxitin in patients with renal insufficiency, which was used as a guideline for determining the dosage of cefoxitin for severely ill patients with renal insufficiency, are included. In addition, a summary of data on file at Merck Sharp and Dohme Research Laboratories (Rahway, New Jersey) that concern 135 bacteremic patients who were treated with cefoxitin and evaluated is presented. Although information derived from septicemic patients is among the most difficult to interpret in regard to the efficacy of an investigational antibiotic, both our experience and the summary of data confirm that cefoxitin is a potent antibiotic suitable for use in septicemic patients when the etiologic agent is susceptible; the data also demonstrate that cefoxitin has a significantly expanded spectrum of activity as compared with that of cephalothin.


Cefoxitin therapy for surgical patients. Cefoxitin was administered to 25 patients on a general surgical service. In 16 of these patients, a bacteriologic diagnosis was made: four patients had peritonitis, one had ascending cholangitis, seven had cellulitis, and four had abscess. The dosage of cefoxitin varied from 4 to 8 g per day and was generally well tolerated, although there were three cases of phlebitis and one superinfection. There was no evidence of renal, liver, or bone marrow toxicity. All isolates of Bacteroides (eight), Escherichia coli (six), and Staphylococcus aureus (five) were sensitive to cefoxitin. Two of three strains of Pseudomonas aeruginosa were resistant. All patients recovered with cefoxitin and corrective surgery. The results indicate that cefoxitin has the potential of replacing combination therapy in some surgical patients.


Clinical evaluation of cefoxitin in the treatment of respiratory tract and other acute bacterial infections. A clinical trial was designed for evaluation of the efficacy, safety, and patient tolerance of cefoxitin. Of 26 patients who could be evaluated clinically, 21 had respiratory tract infections; all but one patient responded satisfactorily to therapy with cefoxitin. There were no adverse systemic reactions, nor was any renal or hematologic toxicity noted. In two instances levels of hepatic enzymes became slightly elevated; these changes were possibly related to administration of cefoxitin. About one-third of patients receiving cefoxitin developed minor local reactions at the site of intravenous administration; however, there was only a single instance of true thrombophlebitis. Results of this study suggest that cefoxitin is a safe and well-tolerated antibiotic that is efficacious in therapy of acute respiratory tract infections caused by bacteria that are sensitive to this drug.


Cefoxitin in obstetric and gynecologic infections. The use of cefoxitin sodium in 178 patients with a wide variety of obstetric and gynecologic infections was studied. Clinical cure occurred in 94% of the study population, and two women with serious underlying medical problems died. These good results with cefoxitin should stimulate prospective studies comparing this drug to currently employed antibiotic regimens.


A comparison of the effects of prior cold incubation on cerebral cortex function in a hibernator (Cricetus auratus) and a non-hibernator (Cavia porcellus)--II. High energy phosphate levels in cerebral cortex slices after in vitro cold incubation. 1. ATP and CP levels were measured in brain slices from golden hamster and guinea pig after varying periods of cold storage and subsequent incubation at 37 degrees C in the presence and absence of K+ salts. 2. ATP and CP levels were maintained at higher levels in hamster tissue. 3. The results are discussed in relation to the ability of a hibernator to transform and transport chemical energy at low temperatures.


Cefoxitin in treatment of obstetric and gynecologic infections. Cefoxitin was found to be an effective and safe agent for the treatment of infections of the female genital tract that were associated with a variety of aerobic and anaerobic bacteria, including Streptococcus, Escherichia coli, and Bacteroides species. The overall clinical response was 91%. The major cause of failure was the presence of an abscess requiring surgical drainage. Both the intravenous and the intramuscular routes of administration for cefoxitin were tolerated well or moderately well by most patients.


Clinical evaluation of cefoxitin in treatment of infections in 47 orthopedic patients. Cefoxitin was administered to 47 patients on an orthopedic service; 1 or 2 g of the drug was given intravenously every 4-8 hr. Thirty-one evaluated patients with acute or chronic infections of bone, joint, or muscle and tendon had an overall rate of cure of 84%. Ten patients with spinal cord injuries who had urinary tract infections due to Serratia or indole-positive Proteus were treated, and all 10 were cured. Significant adverse effects of cefoxitin included one case of fever due to the drug and three cases of superinfection. Cefoxitin therapy was successful in a variety of difficult-to-treat infections, including some of polymicrobial etiology.


Galactose utilization by the rat tapeworm, Hymenolepis diminuta. 1. Hymenolepis diminuta incorporated label from 14C-galactose into glycogen, but the sugar would not support net glycogen synthesis. Glucose stimulated the incorporation of label from 14C-galactose into glycogen, while glycerol did not. 2. During incubations in galactose, large internal pools of galactose and galactose 1-P accumulated, while the concentration of glucose 6-phosphate remained unchanged. 3. In vitro culture experiments indicated that galactose would not support worm growth. Therefore, while galactose can be metabolized to a limited extent, it cannot substitute for glucose as a nutrient source.


Some properties of pancreatic lipase in Salmo gairdnerii Rich.:Km, effects of bile salts and Ca2+, gel filtrations. 1. The Michaelis constant (Km) of the trout pancreatic lipase is 1.3 x 10(-6) M tributyrin, calculated by the interface concentration of the emulsion. This value is lower than that of porcine pancreatic lipase. 2. The lipase hydrolyses tributyrin in a Ca2+ free medium. Conversely, Ca2+ is essential for the lipolysis of triolein. The cation might be an effector of the reaction, but it seems to remove the inhibition of the enzyme by its product. 3. The curves of the lipase activity according to bile salt concentration seem to suggest the existence of a colipase, that we have not evidenced yet by direct procedures. 4. The apparent molecular weight of the lipase seems to be lower in the trout than in the species studied so far.


In vitro activity of cefoxitin and parenterally administered cephalosporins against anaerobic bacteria. The in vitro activity of cefoxitin against 221 recent isolates of anaerobic bacteria was compared with that of cefamandole and cefuroxime by the agar dilution technique. At achievable serum levels of 32 micrograms/ml, all three drugs were active against most groups of anaerobic bacteria tested. Cefoxitin was active against most strains of the Bacteroides fragilis group, whereas cefamandole and cefuroxime were relatively ineffective. Cefoxitin was least active against Clostridium species other than Clostridium perfringens, whereas cefamandole was very active and cefuroxime moderately active. A review of recent in vitro studies with cefoxitin and parenterally administered cephalosporins indicates that cefoxitin is the most active compound against the B. fragilis group and is least active against Clostridium species other than C. perfringens.


The fetid foot: lower-extremity infections in patients with diabetes mellitus. Infections of the lower extremities in patients with diabetes mellitus have been attributed to Staphylococcus aureus and other facultatively anaerobic bacteria. However, a review of 30 consecutive diabetics who required surgery for lower-extremity infections revealed that 17 had mixed infections due to both obligate and facultative anaerobes; only six had infections due to S. aureus. Mixed infections often had characteristics of anaerobic suppuration and responded poorly to therapy. Patients with mixed infections required more operations than did those with staphylococcal infections, and their surgical wounds healed more slowly. Seven patients had infections with mixed enteric bacteria (neither anaerobes nor S. aureus), and their response to therapy was intermediate between that of the patients with mixed anaerobic infections and those with staphylococcal infections. Nine additional patients with mixed anaerobic infections were treated with cefoxitin; three required amputations at a level above the ankle, but six patients improved and required only limited surgery that preserved foot function. Bacteroides fragilis was the anaerobe isolated most often. Cefoxitin (less than or equal to 20 micrograms/ml) inhibited all of the anaerobes isolated from the nine patients and 97% of an additional 135 consecutive clinical isolates of B. fragilis; 98% of 54 strains of Bacteroides oralis and all of 34 strains of Bacteroides melaninogenicus were also inhibited. These in vitro results and the results of our clinical study show that cefoxitin is a promising antibiotic for therapy of foot infections due to a mixed flora of anaerobes in diabetics.


Rat casein: isolation and characterization of two major fractions. 1. In rat milk, casein exists as particles of 77 nm mean diameter, similar in appearance to the casein micelles in the milk of other species. 2. The heterogeneity of rat casein was investigated by polyacrylamide gel electrophoresis and by chromatography on DEAE-cellulose, both in the presence of 8 M-urea. 3. The chromatography yielded two fractions, D2 and D4. 4. In chemical composition, D2 resembles the kappa-caseins of other species, while D4 resembles the alpha 5-caseins of other species. 5. D2 is soluble in 80 mM-CaCl2 at 37 degrees C, but D4 is insoluble under these conditions. 6. In 80 mM-CaCl2 at 37 degrees C, D2 prevents the precipitation of D4 by the formation of particles similar to those found in rat milk.


Enzymes of heterotherms: LDH of hibernating and normothermic little brown bats, Myotis lucifugus. 1. Lactate dehydrogenase (LDH) was isolated from pectoral or flight muscle, liver and heart of hibernating and normothermic bats, Myotis lucifugus. 2. Activities at high substrate concentrations and the interactions between substrate and enzyme were studied to evaluate possible adaptations associated with the extended period of depressed metabolism of this bat. 3. Activity levels in all tissues decreased with hibernation, although the magnitude was dependent upon both temperature and substrate concentration. In general, the extent of decrease was in the order pectoral muscle greater than liver much greater than heart. 4. No correlation was found between the temperature sensitivity of the tissue LDHs as estimated by Ea-values, and the physiological state. 5. However, affinity parameters as estimated by Km-values were less temperature sensitive in hibernator tissues in contrast to the marked perturbations exhibited at temperature extremes in the normotherm enzymes. 6. It is suggested that changes in subunit quantities are in part responsible for these kinetic differences, and not major qualitative isozyme changes. 7. The net result of these effects is a reduced carbon flux to lactate during hibernation, but the maintenance of a potential for LDH catalyses during arousal.


Pharmacokinetics and comparative pharmacology of cefoxitin and cephalosporins. Features of the distribution, metabolism, elimination, and pharmacokinetics of the cephalosporins and cefoxitin must be considered when concentrations of these drugs in biological fluids are interpreted. The extensive (approximately 86%) binding of cefazolin to plasma protein may account for the smaller volume of distribution and slower rate of renal clearance than are observed for cefoxitin, which is less extensively (73%) bound to protein. Results of microbiological assays of drug in urine may be influenced by the extent of metabolism of the drugs, which is 33% for cephalothin but less than 2% for cefoxitin. Elimination of cephalosporins and cefoxitin occurs by both glomerular filtration and tubular secretion and can be inhibited by the concurrent administration of probenecid. The pharmacokinetics of cefoxitin may be described by a linear, two-compartment, open model that has been used to predict levels of drug achieved in serum and urine after various dose regimens, including administration by intravenous bolus or infusion. The bioavailability of intramuscularly administered cefoxitin is equivalent to that of intravenously administered cefoxitin and is 90% complete within 3-4 hr after the dose is given.


Ontogenic development of swine (Sus domesticus) adipose tissue lipases. 1. The in vitro activities of lipoprotein lipase (LPL) and hormone sensitive lipase (HSL) were examined in adipose tissue preparations from pigs 0-150 days of age. 2. The activities of both LPL and HSL increased 3- to 4-fold between birth and day 2 postpartum, remained at relatively high levels through weaning, and fell sharply in the oldest animals (150 days). 3. The decline in enzyme activities at older ages could partially be attributed to an increase in adipocyte size.


scyllo-inositol and myo-inositol levels in tissues of the skate Raja erinacea. 1. Many organs of the skate Raja erinacea have been found to contain scyllo-inositol levels that are much higher than myo-inositol, the opposite to that found in mammals. 2. Both inositols were found in all skate organs studied. 3. myo-Inosose-2 was found to accompany the inositols in many of these organs.


The influence of three days or three weeks of force feeding on the transport of plasma lipids in young female chickens (Gallus gallus L.). 1. Ten-week old broiler females were force-fed (FF) for 3 days or 3 weeks. 2. Control livers were lighter in weight and contained less total lipid, neutral lipid and phospholipid than either FF group, which did not differ. 3. Radioactivity incorporated into liver neutral lipid fractions from 1-[14C]acetate injection was greater in birds FF 3 weeks than controls. Those FF 3 days were intermediate. In all groups, the triglyceride fraction contained 90-94% of isolated radioactivity, the cholesterol fraction 4-8% and the cholesterol ester fraction 1-2%. 4. Plasma lipids were elevated in the birds FF for 3 weeks, but not in those FF 3 days. After injection of 1-[14C]acetate, plasma lipid specific radioactivities were not different between the 3 groups at 20 and 60 min post injection, but were greater in the controls at 120 min.


A comparison of selected physical properties of hepatic sorbitol dehydrogenases [L-iditol: NAD oxidoreductases] from four mammalian species. 1. The sorbitol dehydrogenases [L-iditol: NAD oxidoreductase] from livers of cow, man, rat and sheep each possess molecular weights of about 140,000. The beef, rat and sheep liver enzymes are composed of subunits of molecular weight 40,000. 2. The sorbitol dehydrogenases from livers of these four species each possess an isoelectric point of 7.3. 3. The four enzyme preparations show identical mobilities upon disc-gel electrophoresis and yield a single band of enzymic activity. 4. Sorbitol dehydrogenase activity is activated by the presence of ampholines or by increasing ionic strengths, with maximal activation at about 0.5 M salt concentration. These factors may cause the Km for NAD to be lowered.


Classical and alternate complement pathway activities in paired dairy cow--newborn calf sera. Hemolytic assays were used to compare alternate and classical C pathway activities in sera obtained from clinically normal newborn dairy calves and their mothers at the time of delivery. Mean alternate and classical CH50 concentrations in sera from newborn calves were both significantly lower than in their dams (P less than 0.001). The titer of alternate C pathway activity, expressed as CH50 units/ml, in sera from 17 calves was 12.9 +/- 5.5, whereas for the cows it was 25.8 +/- 6.2. The ratio of cow: calf serum alternate CH50 titers averaged 2.25 +/- 0.80 and ranged from 0.88 to 4.14. Classical CH50 titers were 78.0 +/- 42.7 units/ml in calf sera and 246.0 +/- 44.5 in cow sera. The ratio of cow: calf serum classical CH50 titers averaged 3.71 +/- 1.49 and ranged from 1.19 to 6.87. The wide range of values, noted for both the alternate and classical C pathways, within maternal and neonatal groups was assumed to reflect the biologic variability of complement levels in bovine serum. The possible relationships between deficient levels of alternate and classical CH50 activity in newborn calves and their susceptibility to infections is discussed.


Experimental studies of chronic pneumonia of sheep. Strains of Mycoplasma ovipneumoniae and Pasteurella haemolytica isolated from sheep affected with chronic pneumonia were inoculated by endobronchial route to conventionally-reared and SPF (Specific Pathogen-Free) lambs. Changes resembling those of the naturally-occurring disease were produced in most lambs given the organisms in combination and in some given M. ovipneumoniae alone. Similar but less extensive changes were seen in SPF lambs and fewer animals were affected. Different strains of M. ovipneumoniae did not affect the extent of changes produced in SPF lambs. M. ovipneumoniae became established in the lungs of both types of sheep; P. haemolytica did so less readily. It was concluded that chronic pneumonia may be reproduced in conventional animals by combined inoculation of M. ovipneumoniae and P. haemolytica. Age and status of immunity to mycoplasmas may account for the different responses of conventional and SPF lambs.


[Potency testing of inactivated rabies vaccines. Problems posed by the use of the Habel test]. Rabies vaccines containing inactivated virus are often tested for potency by the Habel test. Even though many people use the Habel test, a humpbacked curve of mortality is always observed for the vaccinated mice, with a linear regression for the control animals. This unexpected survival of vaccinated mice given the greatest doses of virus is an artefact since the phenomenon is not observed after a non intracerebral challenge. Since the intracerebral challenge is not a good model for natural infection, the index of protection for the tested vaccines must be evaluated after discarding aberrant values by a graphic method. The use of the Reed and Muench method or Kärber method leads to an overestimated index of protection. The N.I.H. test gives the best estimation of the effectiveness of rabies vaccines, but does not reproduce the natural conditions of vaccination and infection.


Lobster (Homarus americanus) striated muscle myosin. 1. Myosin from the thin-filament regulated flexor muscle of lobster contains 2 moles of each of 2 light chains. 2. The Lb 1 light chain of 19,000 daltons which can be removed by DTNB is heavier than the DTNB light chain of chicken. The Lb 2 light chain of 17,000 daltons can be removed with urea. 3. On electrophoresis in 8 M urea (pH 8.7) the Lb 2 light chain migrates with a mobility similar to that of chicken A2, but the Lb 1 migrates significantly faster than any of the chicken light chains. 4. In lobster, the DTNB treatment destroys the Ca and K-EDTA ATPase activity of lobster myosin.


Bilirubin metabolism in the spiny dogfish, Squalus acanthias, and the small skate, Raja erinacea. 1. The main bilirubin conjugate in bile of spiny dogfish (Squalus Acanthias) and small skate (Raja Erinacea) is bilirubin monoglucuronide. 2. Microsomal preparations from dogfish and small skate liver have similar bilirubin UDPglucuronyltransferase (UDPGT) activity and catalyze the conjugation of bilirubin with glucose from UDPglucose. 3. The activity of bilirubin glucosidation (UDPGT) was 0.5 times UDPG1T activity in dogfish and 0.15 times in skate liver microsomes. 4. Sodium cholate increased UDPGT and UDPG1T activities in dogfish and skate liver microsomal preparations only minimally, but the detergent markedly increased thermolability of UDPGT in skate liver microsomes.


Effect of photoperiod and cold acclimation upon plasma free fatty acid levels in the white rat. 1. Animals were acclimated at 3 +/- 1 degrees C and at room temperature (22 degrees C) for 3 weeks. 2. At each acclimation temperature animals were maintained under either an 8:16 L:D cycle or a 16:8 L:D cycle. 3. Blood samples were taken before and after exposure to -38 degrees C for 30 min. 4. Free fatty acid levels were greatest in cold acclimated animals which were maintained on a short light cycle. 5. Interaction between acclimation and photoperiod was apparent.


Composition of adrenal lipids from some domestic and wild ruminants. 1. Lipids from the whole adrenal glands of the ox and the African buffalo (Syncerus caffer) were extracted and fractionated into neutral and phospholipids. Both species revealed the presence of considerable quantities of cholesterol but only very small quantities of cholesteryl esters. 2. Fatty acids from various fractions of bovine and buffalo adrenal glands were investigated by gas-chromatography. They showed a remarkably low content of higher unsaturated fatty acids and a very high content of stearic acid. 3. Mitochondrial and microsomal fractions were isolated from the adrenal glands of the impala antelope (Aepyceros melampus), their lipids extracted, analyzed and compared with the composition of the mitochondrial lipids from bovine adrenal glands. 4. Subcellular fractions of the bovine and impala adrenal glands contain only very small quantities of esterified cholesterol. Most of the lipid fractions were characterized by the absence of adrenic acid (C22:4 omega 6) and a low content of arachidonic acid.


Chicken secretory immunoglobulin: chemical and immunological characterization of chicken IgA. 1. Chicken IgA purified from biliary fluids was chemically and immunologically characterized. 2. Chicken IgA was determined to be the only immunoglobulin class present in bile. Gel filtration studies reveal polymeric IgA e.g. 17-19S. 3. Antigenically, chicken IgA is distinct from chicken IgG, and IgM. 4. Chicken IgA does not show antigenic homology to human IgA. 5. SDS poly-acrylamide gel electrophoresis revealed IgA to possess heavy chains of 60,000 and light chains of 24,000 mol. wt, respectively. 6. Peptide mapping of tryptic digests of chicken alpha chains reveals approximately 35 peptides. The peptide map pattern is distinct from chicken gamma chains.


Immunogenicities of haemocyanins from the giant African snail (Achatina fulica) and the keyhole limpet (Megathura crenulata). 1. The immunogenicities of the giant African snail (Achatina fulica) haemocyanin (AFH) and the keyhole limpet (Megathura crenulata) haemocyanin (KLH) were compared by determining their capacities to induce humoral and cell-mediated immune response in rats. 2. KLH was found to be more immunogenic than AFH, and this was attributed to the fact that KLH contains more moieties, and probably more antigenic determinants than AFH. 3. Since AFH was found capable of stimulating both humoral and cell-mediated immunity, it was suggested that it could be used as an antigen for the investigation of immune responses.


Cobalamin binding proteins in stomach and serum from various animal species data for B12 binding capacities and molecular sizes of the binding proteins. 1. The unsaturated cobalamin-binding capacity of stomach mucosa and serum from 37 animals and the size of the binders have been measured. 2. The binding capacity in stomach mucosa was from 1 to 600 nmole kg-1 wet wt, the highest values occurring in pig, guinea pig, porpoise and earthworm. In serum it varied from 0.4 to 270 nmole 1(-1), the highest values occurring in duck, grass snake, toad and spiny dogfish. 3. The size in terms of Stokes radius of the cobalamin binders in stomach mucosa varied from 1 nm in hagfish (Mr approximately 14,000) to 7 nm in lamprey (Mr approximately 210,000). In serum it was from 1.4 nm (Mr approximately 17,000) to 7.4 nm (Mr approximately 240,000) both in frog.


Sources of normal variation of plasma lactate dehydrogenase in range cattle. 1. Plasma levels of lactate dehydrogenase (LD) were estimated in Hereford cattle of 1 month to 12 yr of age kept under range conditions. 2. Plasma levels of LD were estimated in a group of confined cows and their calves fed a constant diet and kept in individual pens in the same geographic area as the range Herefords. 3. Large seasonal changes in mean plasma LD were observed in the range cattle but not in the confined, constant diet group. The seasonal changes corresponded with the appearance of new growth on the range. 4. Changes in plasma LD were observed at weaning time in the range cattle but not in the confined group. 5. Sex had no effect upon plasma LD levels in the juvenile animals of either group. No comparison of adult animals could be made. 6. Mean plasma LD levels did not change with age but the seasonal effect decreased in magnitude as the animals matured. 7. An increase in plasma LD was observed around the first ovulation in the confined animals. 8. A decrease in plasma LD was observed during parturition in the confined group.


A comparative study of haemoglobins from the Artiodactyla by isoelectric focusing. 1. Isoelectric focusing on 7% acrylamide gels in 2% ampholyte of pH range 6-8 was used to fractionate the haemoglobins of 81 animals, representing 30 species from 4 families of the Order Artiodactyla. 2. Isoelectric points of the major haemoglobins were determined and the relative distribution of minor haemoglobins compared. 3. Marked haemoglobin heterogeneity was observed, with polymorphic and multiple haemoglobins. No one haemoglobin was common to all species.


Schistosoma mansoni and Schistosoma japonicum: comparison of glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase activities in adults. Glucose-6-phosphate dehydrogenase activity in paired adult Schistosoma mansoni is about twice as great as in paired adult Schistosoma japonicum. 2. 6-phosphogluconate dehydrogenase activity accounts for 25.8% of the measured production of reduced nicotinamide adenine dinucleotide phosphate (NADPH) in S. japonicum but only 8.6% of the measured production of NADPH in S. mansoni. 3. These data suggest a species difference in 6-phosphogluconate metabolism.


Anserine and carnosine in chicks (Gallus gallus) rat pups (Rattus rattus) and ducklings (Anas platyrhynchos): comparative ontogenetic observations. 1. Muscle and brain from developing chick embryos, as well as from day-old chicks, rats, and ducks were analyzed for the histidine-containing dipeptides, anserine and carnosine. 2. Anserine was found in the brain of all species studied, whereas in muscle, anserine was found only in chicks. 3. At 15 days, the muscle of developing chick embryo contained 41 +/- 9 mumoles/100 g anserine while carnosine was present at a level of less than 3 mumoles/100 g. 4. In day-old chicks the anserine level in muscle was 100 +/- 35 mumoles/100 g while the carnosine level was 22.5 +/- 1 mumoles/100 g. 5. These findings cast doubt on earlier hypotheses relating anserine and carnosine to muscle activity.


Comparative studies on the nonprotein but nondialysable sugar-peptide fraction isolated from the plasma of various classes of vertebrates. 1. The nonprotein but nondialysable ninhydrin-positive substances isolated from plasma of some species belonging to four classes of vertebrates (fishes, amphibians, birds and mammals) were studied. 2. In all species studied (Cyprinus carpio, Salmo gairdneri, Rana esculenta, Rana temporaria, Gallus domestica and Rattus rattus) a similar group of nondialysable ninhydrin-positive substances may be isolated from the plasma by Sephadex G-25 filtration followed by dialysis. 3. Thin-layer Sephadex G-25 electrophoresis disclosed in birds (Gallus domestica) the lack of one ninhydrin-positive band contrary to the species of other classes of vertebrates. 4. Electrochromatography showed in all species studied an alteration in the composition of the basic peptide fractions and in birds the lack of one neutral peptide.


Comparative studies on the glycoprotein composition of mammalian platelets. 1. The membrane glycoprotein composition of the blood platelets of 13 mammalian species has been compared by SDS-polyacrylamide gel electrophoresis. 2. A basic pattern of 2-3 predominant high molecular weight glycoprotein bands was observed, however species differences in their relative rates of migration and abundance were apparent. 3. Wide species differences in the number and rate of migration of the acidic glycopeptides released by trypsin digestion of washed platelet suspensions were observed following polyacrylamide gel electrophoresis in the absence of SDS.


Glucose turnover in the young chicken (Gallus domesticus) using variously labeled (3H, U-14C) glucose tracers. 1. Various parameters of glucose metabolism--glucose replacement rate, percent recycling, mean transit time, and glucose mass were examined using various double labeled glucose tracers--2T-, U-14C; 3T-, U-14C; 4T-, U-14C; 5T-, U14C; and 6T-, U-14C. 2. Estimates of replacement rate were greatest for 2T-glucose (21.4 mg/min/kg), with 3T-, 4T-, 5T-, and 6T-glucose all having similar values (15.8, 15.6, 17.0, 16.0 mg/min/kg, respectively). 3. Calculated glucose mass based on all tritiated tracers (734-1086 mg/kg body weight) agreed closely with a direct determination of body glucose (969 mg/kg). 4. Reincorporation of tritium from 3H2O into glucose did not occur to any significant degree. 5. The young chick was found to have a very rapid rate of glucose turnover and high percent recycling compared to mammals.


Lactic acid formation in crustaceans and the liver function of the midgut gland questioned. 1. The possibility of the midgut gland of the crustacean (Cherax destructor) functioning as a liver has been investigated. 2. Seven species of crustaceans accumulate lactic acid in the haemolymph when exercised. The rate of disappearance of lactate in Homarus gammarus and in C. destructor is very slow when compared with man. 3. In the midgut gland of C. destructor no firm evidence was obtained for gluconeogenesis from lactate and for ketogenesis from fatty acids. 4. It is concluded that there is at present no justification for the common practice of calling the midgut gland an hepatopancreas.


Utilization of a differential hemoglobin elution procedure as a rapid assay for identification of embryonic and adult chicken red blood cells. 1. A hemoglobin elution-staining procedure has been developed for distinguishing embryonic chick red blood cells from adult chicken red blood cells. 2. Adult hemoglobin is eluted from red blood cells with 1.9 M potassium phosphate buffer, pH 7.2; whereas, embryonic hemoglobin is retained within the cells and gives positive staining with erythrosin B. 3. The hemoglobin elution-staining pattern during development can be correlated with two embryonic hemoglobins as detected by polyacrylamide gel electrophoresis. 4. The series of red blood cells staining with erythrosin B correspond to the primary erythrocyte series suggesting that hemoglobin expression during development is correlated with different cell populations.


Temperature induced isozyme variants in individuals of the sea urchin, Arbacia punctulata. 1. The technique of microacrylamide slab gel electrophoresis was used to repeatedly monitor the qualitative isozyme composition in the tube feet of the sea urchin, Arbacia punctulata exposed to different temperature regimes. 2. Four enzyme systems were assayed. Three (MDH, HK, and ACPH) were monomorphic for each animal studied and no change in band migration pattern was observed. Banding patterns for the fourth system (EST) were observed to vary in the same individual. 3. The reversible induction of esterase variants in the tube feet of the same individual is reported. 4. The change was temperature dependent, and a period of 7-14 days acclimation was required to induce the pattern alteration.


Variance in LDH isozyme patterns in a Chinese hamster (Cricetulus griseus) colony. 1. LDH activity and isozyme pattern were examined in the liver and epididymal fat pad of animals in 12 different sublines of the Upjohn Chinese hamster colony, which was established to produce animals with spontaneous diabetes. 2. Considerable divergence was observed and the animals could be divided into 3 groups according to LDH-H activity. Each group was significantly different from the other in epididymal fat pad LDH-1, 2,3 and 5 and liver LDH-3, 4 and 5. 3. The variance in LDH isozyme pattern bore no relationship to the state of diabetes but appeared to arise from other genetic determinants. However, within a single subline, a significant correlation between blood sugar and epididymal fat pad LDH-5 was observed.


Metabolism of oleic, linoleic and linolenic acids in gourami (Trichogaster cosby) fry and mature females. 1. Female gouramis incorporated pulse-fed [U-14C]oleic, linoleic and linolenic acids more readily into roe than body lipids. Labeling was highest in eggs spawned 20-30 days after feeding. 2. In the fry, linoleic and linolenic were catabolized more slowly than oleic acid, indicating conservation of the polyunsaturated acids in the early stage of life. 3. In the mature female, metabolism of linolenic was distinct from that of the other acids by more extensive conversions and greater use of 14C for de novo synthesis of fatty acids.


Plasma lipid levels and lipoprotein ratios in ten rodent species. Cholesterol, triglyceride, and non-esterified fatty acid (NEFA) levels and lipoprotein electrophoretic patterns in blood plasma of ten species of wild rodents are compared with those of the laboratory mouse and man under standard conditions. 1. Average plasma lipid levels for the mouse and man appear in the middle of the wide ranges of 0.6-1.8 g/l. for cholesterol, 0.4 to 2.4 g/l. for triglyceride and 0.32-0.68 m-equiv/l. for NEFA found in these rodents suggesting species may vary in their ability to utilize these lipids. 2. Some universal relationship between levels of cholesterol and triglyceride, but not with NEFA, are suggested by a comparison between average levels in these 12 species. 3. Lack of correlation between the typical four component human lipoprotein electrophoretic pattern with any of the uniquely different patterns of rodents indicate using this method it is not feasible to compare the blood lipid transport system amongst species.


The effects of starvation on the sodium, potassium, water and free amino acid content of parietal muscle from Agonus cataphractus. 1. The levels of the individual free amino acids, K, Na and water present in parietal muscle were measured in fish starved for periods of 3-115 days. 2. The Na and water content progressively increased relative to dry weight of tissue during the starvation period whereas an initial increase in the K content up to 67 days was followed by a decrease to near the normal fed level by 115 days. The ammonia content remained essentially stable over the same period. 3. The essential free amino acid content tended to follow the muscle sodium pattern while the non-essential amino acids and taurine gave results somewhat similar to those of potassium. 4. The net result of these changes is that the Na, K and total free amino acid concentration is maintained within fairly narrow limits relative to the tissue water content during 11-67 days of the starvation period under the experimental conditions used.


Regulation of purine metabolism: a comparative study of the kinetic properties of adenylosuccinate synthetases from various sources. 1. Adenylosuccinate synthetase has been partially purified from rat liver, fetal rat liver, Novikoff ascites cells, Walker carcinoma 256 solid tumors, chicken liver and muscle, rabbit muscle and pig brain. 2. Considerable differences exist in Michaelis constants among the various species and the changes possibly reflect differences in regulation. 3. The kinetic properties of the enzyme are generally consistent with proposed metabolic roles in various tissues.


Expression of serum proteins in the developing chick embryo. 1. Sera from different embryonic stages of development and adult chickens were examined for embryonic-adult differences. 2. An antisera prepared in rabbits to 14 day embryonic chick sera revealed an antigen present in the early stages of embryonic development. The antigen reached a peak on day 14 and was not detectable 5 days post hatching. 3. Gel filtration studies in conjunction with immunodiffusion studies revealed the serum antigen to be in the 160,000 mol wt range. 4. Immunoelectrophoretic analyses of embryonic and adult sera revealed the sera components to increase in quantity and in numbers with development. The serum immunoelectrophoretic profile of newly hatch chick serum closely resembled sera from adult chickens. 5. Polyacrylamide gel electrophoretic comparisons of the sera of the developing chicks and the adult chickens revealed proteins specific to certain developmental stages as well as proteins specific to the adult serum. Qualitative and quantitative serum protein differences were found.