[An experimental study on Pseudomonas osteomyelitis with special reference to the production of experimental osteomyelitis in mice (author's transl)]. I) The author has successfully produced a model of experimental osteomyelitis caused by pseudomonas aeruginosa using the following procedure though such a demonstration has been said to be very difficult. After impregnation in a solution containing about 10(5) pseudomonas aeruginosa, 3 mm silk thread of No. 5 was dried under low-pressure atmosphere and then inserted into the metaphysis of right tibia of a mouse. This method can be produced experimental osteomyelitis in 100% of the animals. In the experimental osteomyelitis generated pathologically by this method, inoculated organisms do not transmigrate into blood, the kidney and the contralateral tibia. This may therefore be regarded as a local infection causing no death, making a long period of observation possible. In view of the X-ray and patho-histological findings, it is similar to human osteomyelitis. Furthermore, its host is a pure-bred mouse with constant elements making a league-scale experiment possible. II) This is an experimental model of osteomyelitis proved quite useful for the quantitative analysis of the effects of antibiotics, and would be a good method for evaluation of antibiotics to be developed in the future.


Systematic multi-modal treatment of osteosarcoma, with special reference to the role of fast neutron radiotherapy. Sixty-one patients with osteosarcoma were treated. Twenty-four of these patients were managed with our systematic multi-modal treatment. Overall survival rate was markedly improved chiefly by intensive systemic chemotherapy with multiple drugs, especially adriamycin and high dose methotrexate. Preoperative regional intra-arterial infusion of anti-tumor drugs and fast neutron radiotherapy were employed and it was suggested that fast neutron had a higher relative biological effectiveness and a greater therapeutic gain factor as compared with X-rays. Fast neutron radiotherapy can play a significant role in the very systematic treatment of osteosarcoma and is specifically useful for preservation of the affected limbs.


Experimental intra-oral carcinoma.

Experimental intra-oral carcinoma. Fundamental studies of tumour host relationships in oral cancer cases would be assisted if a suitable animal model could be found. Current concepts in the use of animal models for the study of intra-oral carcinoma are reviewed and the development of a unique transplantable tumour system which had been induced and maintained only in syngeneic animals, is recorded.


Effects of methamphetamine and scopolamine on variability of response location. Methamphetamine and scopolamine were studied in monkeys responding under a multiple fixed-ratio fixed-interval schedule of reinforcement. A response on any one of six levers could satisfy the schedule requirements. Variability of response location was evaluated in terms of switches, where a switch was defined as a response on one lever followed by a response on a different lever. Under baseline conditions the fixed-ratio schedule generated a high rate of responding and a low level of variability, while the fixed-interval schedule generated a low rate of responding and a high level of variability. Both methamphetamine (0.1 to 0.5 mg/kg) and scopolamine (2.4 to 240 microgram/kg) decreased overall response rate and increased variability of response location in each component of the multiple schedule with increasing doses of drug. At lower doses both drugs were found to decrease rate without affecting response variability.


Distinct Ir genes for helper and killer cells in the cytotoxic response to H-Y antigen. The H-Y-specific cytotoxic T-cell response requires helper cells: cells from bone marrow chimeras B6 X CBA leads to B6, B6 X CBA leads to B10.A (5R), or B6 X CBA leads to CBA are each unable to respond to H-2k male cells. If, however, cells from B6 X CBA leads to B6 or B6 X CBA leads to B10.A (5R) chimeras are adoptively transferred together with cells from B6 X CBA leads to CBA chimeras, H-Y-specific CTL restricted to H-2k can be obtained. Thus, cells from B6 X CBA leads to B6 or B6 X CBA leads to B10.,A (5R) chimeras (restricted to the left end of the H-2b haplotype) can help CTL precursors from B6 X CBA leads to CBA chimeras (restricted to H-2k). The two classes of T cells required for the CTL response to H-Y antigen are controlled by different IR genes. All H-Y-specific CTL obtained from chimeras B6 + CBA leads to B6 X CBA were found to be of B6 origin. This suggests that CTL or their precursors must express antigens encoded in the left end of the H-2b haplotype for interaction with helper cells.


The specific binding of Listeria monocytogenes-immune T lymphocytes to macrophages. I. Quantitation and role of H-2 gene products. A system was developed to study the binding of Listeria monocytogenes-specific T cells to L. monocytogenes-pulsed macrophages as an analogue of the initial phase of T-cell activation: antigen recognition. Specific binding, demonstrable after a brief (1 h) contact, was quantitated by the depletion of L. monocytogenes-specific T-cell activity in the cells nonadherent to L. monocytogenes-pulsed macrophage monolayers. L. monocytogenes-specific T-cell function was measured by its ability to activate L. monocytogenes-pulsed macrophages, both to secrete a protein mitogenic for thymocytes and to effect nonspecific tumoricidal activity. These manifestations of T-cell function are known to be regulated by products of I region of the H-2 gene complex. Studies designed to determine the role of H-2 gene products in specific T-cell-macrophage binding have revealed the following. T cells bind specifically to syngeneic macrophages and poorly to allogeneic macrophages. The binding ability appears to map to the K end of the H-2 gene complex (K through I-E). At least two distinct populations of B6AF1 T cells with binding avidity for L. monocytogenes presented on parental macrophages can be identified. Finally, the binding of a given parental-reactive B6AF1 T-cell clone can be specifically inhibited by pretreatment of the antigen-pulsed B6AF1 binding macrophage with anti-H-2 (anti-Ia) antibodies reactive with the appropriate parental haplotype. These results strongly suggest that H-2 gene products play a direct role in mediating the specific binding of T cells to macrophages and imply that the antigen-dependent physical interaction between T cells and macrophages is the initial, and determining, event in some forms of H-2 gene control of immune reactivity.


Patterns of virus-immune T-cell responsiveness. Comparison of (H-2k X H-2b) leads to H-2b radiation chimeras and negatively selected H-2b lymphocytes. Negatively selected H-2K(b)D(b) TDL can be induced to respond strongly to vaccinia virus presented in the context of both H-2K(k) and H-2D(b) when stimulated in irradiated H-2K(k)D(b) recipients. Addition of excess (H- 2K(k)D(b) x H-2K(b)D(b))F1 TDL, which are low responders to H-2D(b)-vaccinia virus, does not obviously suppress the reactivity pattern of the H-2K(b)D(b) T cells. However, lymphocytes from chimeras made by reconstituting H- 2K(b)D(b) mice with (H-2K(k)D(k) x H-2K(b)D(b))F(l) bone marrow cells make little, if any, cytotoxic T-cell response to vaccinia virus when sensitized in H-2K(k)D(b) recipients. We have thus documented one instance where the responder phenotype of T ceils from an F(l) {arrow} parent chimera is not equivalent to that associated with the H-2 type of the parental thymus. Lymphocytes from both the chimera and the H-2K(b)D(b) parent (after negative selection) are tolerant to the H-2K(k) and I-A(k) alloantigens encountered in the recipient, but the chimera T cells are also defective in their response to a neoantigen (vaccinia virus) presented in the context of H-2K(k) which the parental T cells invariably recognize. It is thus possible that at least part of the phenomenology associated with the F(l) {arrow} parent radiation chimeras reflects deletion of repertoire in the context of H-2 antigens present during thymocyte ontogeny on other than radiation-resistant thymic epithelium.


Ir gene controlled carrier effects in the induction and elicitation of hapten-specific delayed-type hypersensitivity responses. The genetic requirements of carrier recognition were examined in the priming and elicitation of hapten specific, T-cell mediated, delayed-type hypersensitivity (DTH) responses. It was shown that nitrophenyl acetyl-poly-(L-glu56-L-lys35-L-phe9) (NP-GLO) could prime for NP responses only in strains of mice which are Ir gene responders to GLO. In contrast to this requirement, NO-GLO could elicit an NP-specific response in NP-bovine gamma globulin primed mice, even in GLO nonresponder strains. Furthermore, the nonimmunogenic molecule, NP-GL, could elicit an NP-specific DTH response in animals primed with NP on an immunogenic carrier.


Protein synthesis in Bunyamwera virus-infected cells. In Vero cells infected with Bunyamwera virus there is a rapid inhibition of cell RNA and protein synthesis to levels of 30 and 3% respectively of the control rate, both the rate of inhibition and the time lag before its initiation being multiplicity dependent. Using u.v.-irradiated virus, investigation of the mechanism of inhibition of host cell protein synthesis indicates that synthesis of new virus components is required for inhibition to occur. Quantitative comparison of the proteins synthesized in infected cells shows that at higher m.o.i. synthesis of virus, as well as cellular proteins, is inhibited. Bunyamwera virus-infected Vero cells synthesized three virus-specific proteins identified as the structural virion proteins. Nucleoprotein is synthesized predominantly early in infection while the major envelope glycoprotein and the minor glycoprotein are synthesized predominantly late in the infection cycle.


An ethological perspective on social behavior in long stay hospitalized psychiatric patients. Ethological methods were employed to gather normative data on social behavior in long stay male inpatients in the ward environment. Most were diagnosed schizophrenic. Social bheavior was categorized into four main types: altruistic, assertive, cigarette, and verbal. Patients (N = 24) were divided into three groups on the basis of the amount of social interaction (top, middle, and bottom thirds of the interactional scale). With the use of these divisions, results indicated: a) marked variability between thirds in the amount of social behavior exhibited; b) differences in the kinds of social behavior utlized by each third--the top was predominantly verbal whereas the middle and bottom were predominantly nonverbal; c) differences in the modes of interaction between thirds--verbal behavior characterized interactions between the top and top, and top and bottom, whereas assertive behavior characterized interactions between the top and middle, middle and middle, middle and bottom, and bottom and bottom; and d) correlative relationships between various social and nonsocial parameters, the most noteworthy including the positive relationship between head up, eyes open, and high rate of social interaction, and the positive relationships between altruistic, cigarette, and verbal behavior. Discussion focused on issues of variability between patients, stability of behaviors over time, behaviors indicative of bonding and social attentiveness, profiles of behavior characteristic of each third of the interactional scale, and the ethological constructs of dominance and attention structure.


An ethological analysis of long stay hospitalized psychiatric patients. Senders and receivers in social interaction. Ethological methods were employed to monitor social interaction among 24 long stay male patients on a psychiatric ward. Most were diagnosed as schizophrenic. Analysis was made using four molar behavioral groupings: assertive, altruistic, cigarette, and verbal. Within each grouping social behavior was divided into two categories: sending and receiving. Patients were ranked in an interactional scale and divided into three groups on the basis of the amount of social interaction: top, middle, and bottom third. Results showed that individuals in the top third tended to be senders of social behavior, whereas individuals in the middle and bottom thirds tended to be receivers. Individuals in the middle received relatively large amounts of assertive behavior. In addition, correlative relationships indicated positive associations between send and receive verbal and send and receive cigarette. No associatiociation existed between send and receive verbal and send and receive assertive. Findings are discussed in terms of a) sending and receiving profiles characteristic of each third of the interactional scale; b) the value of dividing social behavior into components of sending and receiving; and c) the role assertive behavior plays in the social organization of hospitalized groups.


Lyophilised dura mater: experimental implantation and extended clinical neurosurgical use. The historical development of dural substitutes and the process of regeneration of dura mater are reviewed. Lyophilised human cadaver dura mater has been implanted intracranially in baboons and the graft shown to be incorporated with vascularisation but with ossification. In the human, lyophilised dura mater used as a dural substitute also becomes a viable tissue but without ossification. A retrospective study of its use in 100 neurosurgical patients showed a low complication rate and it is suggested that there are occasions when the ready availability of lyophilised dura mater, without the need for a further incision, makes it the dural substitute of choice.


Luxotonic responses of units in macaque striate cortex. 1. Single units in striate cortex were studied in alert macaques while they viewed a ganzfeld. Of the 385 well-isolated units studied for 10 min to 2 h, 24% gave "luxotonic" responses, i.e., their rate of discharge for 1 min or more in diffuse, featureless, wideangle illumination (20-450 cd/m2) was at least double that during a comparable period in darkness, or vice versa, and not attributable to eye movements of blinking. Those discharging faster in the light, "photergic" units, outnumber those responding to darkness, "scotergic" units 1 by 4:1. 2. In the lateral geniculate nucleus, on the other hand, among 46 units studied, 28% were luxotonic, but scotergic units were the more common. Both types were present in both magno- and parvocellular laminae. 3. For striate cortex two-thirds of the luxotonic units were binocular. Some showed highly similar response for either eye alone, and essentially no summation binocularly; others had grossly differing responses from each eye, and complex binocular interaction. 4. Many units of all types at striate cortex showed significant modulation of their activity consequent to saccadic eye movements made in darkness, whereas comparable modulation was not observed at the lateral geniculate nucleus. 5. On the basis of these and other findings it is concluded that luxotonic cortical activity is prominent probably only in alert primates, and that this is a consequence of the fact that all retinal ganglion cells in primates synapse in the lateral geniculate nucleus (Ref. 9). Possible functions range from mere trophic input to providing a veridical image or a scaling factor for maintenance of perceptual constancy in the face of varying levels of general illumination.


Experimental delayed radiation necrosis of the brain. Part 1: Effect of early dexamethasone treatment. The authors have designed an experiment to detect a hitherto unrecognized interaction between high doses of the glucocorticoid, dexamethasone, and brain irradiation. Eighteen juvenile male rhesus monkeys received 1800 rads to the whole brain in 8.5 minutes. For 1 1/2 days before and 10 1/2 days after the irradiation, nine animals received approximately 2.9 mg/kg/day of dexamethasone intramuscularly in addition to irradiation, while the remaining nine animals served as the control group and received saline. All animals eventually developed a progressive neurological syndrome, and died of delayed radiation necrosis of the brain. The two groups were compared with regard to latency to onset of clinical signs, survival time, and number, distribution, and location of lesions of radionecrosis. Large doses of desamethasone did not alter the susceptibility of the primate brain to delayed radiation necrosis. Detailed morphological study of the radionecrotic lesions supports the hypothesis that most, if not all, of the lesions develop as the consequence of injury to blood vessels.


Experimental acute balloon compression of the spinal cord. Factors affecting disappearance and return of the spinal evoked response. Acute balloon compression of the thoracic spinal cord for 15, 7, 5, 3, and 1 minute in monkeys caused immediate disappearance of the spinal evoked response and complete focal ischemia of the compressed segment in all animals. Only the animals in the 1-minute group, however, demonstrated return of the evoked response. These data, coupled with data from previous experiments of slow balloon compression of the spinal cord and spinal cord ischemia, suggest that the major pathological substrate for neural dysfunction after balloon compression of the spinal cord, be it acute or slow, is physical injury of the neural membrane, irrespective of blood flow changes. These findings also suggest that the ability of that membrane to recover is related to rapidity and length of time of compression. Focal changes in blood flow do not appear to be significant in this mechanism.


The effect of early postnatal acquired malnutrition on intestinal growth, disaccharidases and enterokinase. The effect of undernutrition on rat small intestine during the critical newborn period was studied. A severe state of protein-energy malnutrition was induced by litter expansion which caused the mean total body weight of experimentally malnourished rats to diminish significantly as compared to control animals. Intestinal weight and total DNA were similarly diminished in the malnourished rats. DNA and protein expressed per gram wet tissue showed no significant differences between groups. Retarded intestinal growth in the malnourished animals was the result of reduced cell number. The mean specific activities of sucrase and maltase were diminished in the experimental group, with mean activities being 20 to 50% of controls, respectively. These differences were larger when expressed as total organ activities. On the other hand, specific lactase activity was significantly higher in undernourished rats but total lactase activity per organ was similar in both groups. Enterokinase specific activity or total organ activity was significantly higher in the undernourished rats.


Long-term effects of early undernutrition and environmental stimulation on learning performance of adult rats. Early undernutrition was achieved in rats by increasing litter size; the undernourished groups were reared from birth to 21 days in litters of 18 pups per dam and the control groups in litters of six pups per dam. Each of these groups was divided at weaning into two sub-groups: previously undernourished, stimulated (PUS+) and non-stimulated (PUS-); and control, stimulated (CS+) and non-stimulated (CS-). From weaning, all animals were fed ad libitum. Environmental manipulation began at weaning and lasted 11 weeks. "Stimulated" rats were maintained in colonies of three per cage, were handled for 5 minutes each day and allowed to explore a "stimulation chamber" for 30 minutes. All groups were tested in a Hebb-Williams maze at 100 approximately 10 days of age. The nutritional treatment did not impair Hebb-Williams performance, and environmental stimulation improved the learning performance both in control and previously undernourished groups. Neither nutritional nor stimulation treatment affected intraproblem learning. Total DNA and cholesterol were reduced in forebrain and cerebellum in undernourished rats. Brain stem was the region of the CNS least affected by early undernutrition. Environmental manipulation did not modify neurochemical parameters.


Effects of low dietary protein concentration and energy deprivation on lymphocyte transformation in melanoma-bearing mice. Temporal changes in lymphocyte blastogenesis were studied using spleen cells from syngeneic melanoma-bearing and control mice fed various levels of purified diets containing 6, 10 or 30% casein. T-cell blastogenesis was stimulated by the presence of the tumor and these responses changed with the duration of feeding. In addition, protein concentration did not affect T-cell transformation but the level of energy intake influenced concanavalin A induced DNA synthesis. In contrast, the growing melanoma did not influence B-cell transformation whereas a very low level of dietary protein, a low level of energy intake and duration of the dietary manipulation influenced these cells. Tumor weights were generally not affected by the diet except in mice receiving a very low level of energy intake. Thus, we have found that B-cell responses were affected more than those of T-cells and that moderate protein deficiency did not enhance cellular immune responses in syngeneic tumor-bearing and control mice.


Production and metabolism of thyroid hormones in protein-deficient and food-restricted pregnant rats. Thyroxine (T4) and triiodothyronine (T3) concentrations in the plasma and their metabolic clearance and production rates were studied in pregnant protein-deficient and food-restricted rats on the 20th day of gestation. Total T3 levels in the plasma were significantly reduced in malnourished dams, with those of food-restricted dams being consistently lower than those of protein-deficient dams. There were no changes in plasma total T4 concentrations as a result of dietary treatment. Unbound T4 and T3 in the plasma were significantly reduced in both protein-deficient and food-restricted dams. Maternal protein deficiency significantly lowered fetal T4, whereras food restriction caused an elevation. Metabolic clearance rates of T3 and T4 and production rate of T4 were unaffected by diet, while the production rate of T3 was significantly increased in protein deficient dams. Thyroxine to triiodothyronine ratios in malnourished dams were elevated suggesting depressed peripheral conversion of T4 to T3. Alternative factors which may lead to these effects are discussed.


Effect of maternal protein and/or energy deficiency during pregnancy on catecholamines and serotonin in fetal rat brain. An investigation was made of the effect of maternal protein and/or energy deficiency during pregnancy on developmental changes in the levels of catecholamines and serotonin in fetal brain. Pregnant rats were fed on a 20%, 6% or 0% casein diet from day 1 of pregnancy to the day of autopsy (day 18, 20 or 22 of pregnancy). In the control group, the catecholamine content of the brain increased during pregnancy, being 21 ng on day 18, 48 ng on day 20 and 52 ng on day 22. A similar increase was found in the group on a 6% casein diet. In contrast, with complete protein deprivation there was no developmental increase in catecholamine. A slight increase in serotonin and a marked increase in 5-hydroxyindole acetic acid occurred during late pregnancy, irrespective of the maternal diet. At term, the levels of norepinephrine and dopamine and the tyrosine hydroxylase activity in the forebrain, cerebellum and brain stem of the fetuses in the group on a 0% casein diet were significantly less than those in the groups on 20% and 6% casein diets. The free tyrosine concentrations (mumol/g) in the brain of fetuses in the groups on 20%, 6% and 0% casein diets were 0.701, 0.213 and 0.661, respectively. From the above results it is concluded that the low catecholamine content of the brain in fetuses in the group on a 0% casein diet was due to disturbance of the system for catecholamine synthesis, rather than to deficiency of precursors.


The effect on bone repair of aspirin cones placed in extraction sockets in dogs: a histopathologic study. Four canines of eight 2-year-old dogs were extracted surgically, a mucoperiosteal flap being raised. A standardized pair of the post-extraction cavities were filled with three aspirin cones, the other pair being used as controls. The flaps were sutured with surgical gut. The dogs were killed at intervals of 2, 4, 6 and 8 weeks; bone blocks including the cavities were resected and decalcified for histologic examination; serial sections were stained with hematoxylin and eosisin. Microscopic observation on 2 week samples showed an inflammatory response with granulation tissue in both experimental and control cavities with delayed bone apposition in cavities filled with aspirin cones. The 4, 6 and 8 week samples did not show any major difference between the experimental and control cavities. We observed a decrease in infiltrating of cells followed in both by a similarly increasing amount of fibrous tissue and bone remodeling. These results were due perhaps to the details of the experimental method as the suture allowed aspirin to stay in the cavities for only a short time (2--3 days), as occurs in clinical practice.


Focal epithelial hyperplasia in rabbit oral mucosa. Several papules on the floor of the mouth and the ventral surface of the tongue of a zinc-deficient rabbit were studied by light and electron microscopy. These papules appeared to be consistent with focal epithelial hyperplasia of man and chimpanzees. There was a concurrence of cytoplasmic vacuolization with the appearance of viral particles within the nucleus. Intranuclear crystalline viral aggregates were frequently observed in the upper spinous cells.


Relationships among fluoride concentration in enamel, degree of fluorosis and caries incidence in a community residing in a high fluoride area. The caries inhibitory effect of optimally fluoridated drinking water is well documented. Communities are often exposed to naturally fluoridated water with F content exceeding the optimal level. The purpose of this study was to determine the relationship between enamel fluoride, degree of fluorosis (DEGF) and caries incidence (DMFT) in an endemic fluoride area. Eighty-five 14- to 16-year-old children (37 males and 48 females) who were born and had been living continually in the town of Kenhardt in South Africa were selected for this study. The fluoride content of the common water supply is 3.2 ppm F. The DMFT and DEGF were determined clinically. Acid etch biopsies were carried out on both maxillary central incisors of each subject. Ca and F analysis were carried out and mass enamel expressed in microgram and mass F in pg. Data transformations to loge mass F (1nF), loge mass En (1nEn) and square root DMFT were made. The 1nF values were corrected to a standardized depth of 10 micrometer (208.6 microgram enamel equivalent mass) by covariance analysis. Correlations between the parameters square root DMFT, 1nF ad DEGF were calculated, adjusting for the sex of the individual. The correlations were as follows: (formula: see text). A positive association among the three parameters was established.


Cryosurgery of major and minor salivary gland: a light microscopic evaluation in the Rhesus monkey. The minor salivary glands in the hard palate and the submandibular gland of the Rhesus monkey were subjected to a liquid nitrogen, direct-spray, one-cycle freeze. A 4-min freeze in the temperature range of -40 degrees C to -60 degrees C was used. The extent of tissue destruction was determined at intervals up to 1 year. Clinically relevant findings resulted and form the basis for subsequent studies by this group of investigators.


Plasma carnitine levels during intravenous feeding of the neonate. The premature infant has a limited capacity for fatty acid oxidation. This study shows that solutions commonly used for intravenous feedings in the newborn infant contain no carnitine. Infants maintained on this solution have significantly lower total, free, and acylcarnitine levels as compared to when they are fed orally with expressed human milk or a proprietary formula, which is known to contain carnitine. The exogenous supply of carnitine to the premature infant may have a significant influence on the ability to stimulate optimal fatty acid oxidation.


Specific and sensitive high-performance liquid chromatographic determination of glyburide. A specific and sensitive high-performance liquid chromatographic method has been developed for the rapid determination of intact glyburide in dog serum. With butylparaben as an internal standard, 1 ml of acid-buffered serum was extracted with toluene and an aliquot of the toluene was evaporated to dryness. The redissolved residue was chromatographed on a microparticulate reversed-phase column, and quantitation was achieved by monitoring the UV absorbance of the eluate at 228 nm. The response was linear, and the lower detection limit was approximately 20 ng/ml. Assay precision, as estimated by analyzing replicate samples of a laboratory standard, was better than 6% (CV). The utility of the analytical methodology for the determination of this highly potent sulfonylurea in pharmacokinetic studies in the dog was demonstrated.


Urinary metabolites of 3a,4,5,6,7,7a-hexahydro-3-(1-methyl-5-nitro-1H-imidazol-2-yl)-1,2-benzisoxazole in the dog. The antiprotozoal drug 3a,4,5,6,7,7a-hexahydro-3-(1-methyl-5-nitro-1H-imidazol-2-yl)-1,2-benzisoxazole (I), which exhibits activity against trypanosomiasis, is also antibacterial in vivo. Since the urine from a dog dosed with I showed a broader spectrum of antibacterial activity than I itself, metabolites from this urine were isolated and partially characterized. The metabolites were mono- and dihydroxy-substituted species with the hydroxyl groups on carbons 4--7 of the hexahydrobenzisoxazole ring. These observations led to the synthesis of several such hydroxy derivatives of I, and their properties fully supported the proposed positions of metabolic hydroxylation. One synthetic compound, the 6,7-cis-dihydroxy compound, exhibited higher antibacterial activity against Salmonella schottmuelleri in mice and greater trypanocidal activity in vivo against Trypanosoma cruzi (Brazil strain) than I.


Loss of nitroglycerin from aqueous solution into plastic intravenous delivery systems. The mechanism of potential loss of nitroglycerin stored in plastic and glass containers was studied from an equilibrium and kinetic approach. Plastic strips equilibrated with dilute aqueous solutions of neat nitroglycerin showed that the drug was lost by absorption. Drug loss was followed by an electron-capture GLC assay. The same assay of control solutions in glass showed no drug loss in 48 hr at pH 5.7. The kinetics of nitroglycerin absorption and desorption were determined using synthesized 14C-labeled drug. Absorption can be quantified using a diffusion model, where the concentration in the aqueous phase falls with time. Curve fitting yielded an average diffusion coefficient in plastic of 2.05 x 10(-9) cm2/sec and a partition coefficient of 104 (plastic-water) at 30 degrees. Temperature-dependence studies of absorption showed that the diffusion coefficient followed an Arrhenius relationship with an energy requirement of 19.6 kcal/mole, whereas effects on the partition coefficient were negligible. Nitroglycerin desorption from plastic disks under sink conditions into water can be quantified by assuming a diffusion model where the concentration at the surface of a plane sheet remains constant. Nonlinear least-squares curve fitting generated a diffusion coefficient of 1.14 x 10(-9) cm2/sec for the desorption process at 30 degrees.


Influence of dietary protein on the anti-inflammatory and ulcerogenic effects and on the pharmacokinetics of phenylbutazone in rats. Influence of dietary protein deficiency on the anti-inflammatory and ulcerogenic effects and on the kinetics of phenylbutazone was studied in male Sprague-Dawley rats fed ad libitum a 21% (control) or a 5% (low) protein diet for 3 weeks. A low protein diet fed to a decrease in body weight gain, plasma proteins, albumin, globulins, hepatic total and microsomal proteins and in cytochrome P-450. Phenylbutazone produced a greater ulcerogenic effect in rats fed a low protein diet than in control rats; its anti-inflammatory effect did not increase. Plasma t 1/2 of phenylbutazone was longer in protein-deficient rats than in control rats. Dietary protein deprivation led to a decrease in the plasma clearance and plasma protein binding of phenylbutazone but did not lead to a change in its bioavailability. No relationship between the severity of gastric ulceration and the concentration of phenylbutazone or oxyphenbutazone in the stomach tissue was found in any animal of the two groups. The increased susceptibility of protein-deficient rats to the ulcerogenic effect of phenylbutazone was reversible and was not observed when these animals were fed a control diet for 3 weeks. It is concluded that a dietary protein deficiency increases the ulcerogenic toxicity of phenylbutazone relative to its useful anti-inflammatory effects.


Correlation between response to norepinephrine and removal of 45Ca from high-affinity binding sites by extracellular EDTA in rabbit aortic smooth muscle. Divalent ion chelators (EDTA, EGTA) were found to remove 45Ca from high-affinity binding sites in isolated rabbit aortic smooth muscle in a concentration-dependent manner. Measurement of 14C-labeled EDTA uptake (10 to 60-min incubation periods) yielded tissue/medium ratios equivalent to the extracellular space [14C]-sucrose tissue/medium ratio). Addition of high EDTA concentrations (0.5--1.5 mM) elicits large but reversible increases in 45Ca efflux, rapidly removes virtually all 45Ca from muscles previously incubated with 45Ca for as short a time as 7 min (filling of extracellular and superficial sites) or as long as 180 min (more complete equilibration of 45Ca), and blocks the norepinephrine-induced contractile response (this response is only partially inhibited by 0.05 mM EDTA, a concentration too low to remove most of the 45Ca present). Furthermore, phosphatidyl serine, a compound known to decrease exchangeability of 45Ca in this tissue, inhibits the EDTA-induced increase in 45Ca loss. Thus, those slowly depleted Ca++ stores that are important for the contractile action of norepinephrine are removed by EDTA even though this chelator is confined to the extracellular space. Possibly, release or removal of high-affinity Ca++ is regulated by EDTA-accessible Ca++ bound at relatively superficial membrane sites.


Establishing a free-ranging breeding colony of rhesus monkeys. Part I. The Caribbean Primate Research Center recently contracted with the Food and Drug Administration to establish a free-ranging island breeding colony of rhesus monkeys. The goal of the program is to produce 600 to 800 offspring yearly from 1,000 breeding age females. The initial colony stock will consist of approximately 360 animals from an existing colony that was established on an island off the southwestern coast of Puerto Rico in 1961. Expansion of the colony will be accomplished by the purchase and introduction of rhesus females obtained from the wild. The colony site, reproductive history, composition, and the methodology of our expansion plans are discussed. In addition, anticipated problems are identified and analyzed.


The effects of gonadectomy and oestradiol-17 beta on pituitary responsiveness to LH-RH in the adult male marmoset (Callithrix jacchus). Exogenous luteinizing hormone-releasing hormone (LH-RH) administered in a wide range of doses (0.2-25 micrograms) to intact male marmoset monkeys induced a marked increased in plasma luteinizing hormone (LH) concentrations. Maximum LH concentrations achieved after injection of LH-RH occurred progressively later as the dosage increased. Bilateral orchidectomy sigificantly enhanced pituitary responsiveness to a standard dose (2.0 microgram) of LH-RH, whereas the introduction of oestradiol-17 beta implants effectively inhibited the responses. LH-RH-induced LH release after gonadectomy (with and without oestradiol-17 beta treatment) was similar in males and females. The use of marmosets for appropriate investigation into the physiological role of LH-RH in controlling LH secretion in primates is proposed.


Distribution of egg deposits and gross lesions in nonhuman primates infected with Schistosoma haematobium (Iran). Widespread tissue egg deposits and gross lesions were recorded in 15 species of primates subsequent to Schistosoma haematobium (Iran) infections of variable intensity and duration. Considerable extra-intestinal involvement as well as pathology in different parts of the urogenital system were observed. Transitional cell carcinoma of the urinary bladder was recorded for Cebus apella.


Seasonal variation of reproductive parameters in the laboratory-housed male cynomologus macaque (Macaca fascicularis). Changes in body weight, sperm concentration and testicular volume were monitored for a 13-lunar month period in seven laboratory-housed adult male Macaca fascicularis. No significant change was noted in body weight or sperm concentration between periods of maximal and minimal mean values nor between periods standardized for comparison between the three parameters. Testis volumes were significantly greater during the months of July through early September and during the standardized period of May through July than during January through March and the standardized interval October through January.


Clinical appearance of the hyaloid artery system in the newborn rhesus monkey. Fifteen healthy newborn rhesus monkeys (Macaca mulatta) were given funduscopic examinations as part of a screening process before utilization in a research project. The hyaloid vasculature, a system that supplies blood to the embryonic ocular structures, was observed in each subject and was noted to regress clinically at two to three weeks of age. Although the phenomenon has been described morphologically in the literature, its clinical appearance has not been illustrated for the primate clinician.


An X-ray cephalometric analysis of premaxillary growth in operated and unoperated baboons (Papio cynocephalus). Rates of growth of the premaxilla in baboons, which have undergone removal of the nasal septum, and unoperated controls are compared using X-ray cephalometric analysis and direct measures. It is found that our X-ray analysis without the use of metallic implants is not sensitive enough to detect changes of growth rate which are detectable through direct measurement.


Sugar hydrolases and their arrangement on the rat intestinal microvillus membrane. The arrangement of the sugar hydrolases, sucrase-isomaltase, maltase, and lactase on the microvillus membrane of rat intestine was investigated by immunological technique. The enzymes were purified essentially free of each other to near homogeneity and antisera of high specificity were obtained against each. Microvillus membranes were prepared routinely in high purity from rat intestine and contained an average 61% protein, 20% lipid, and 19% carbohydrate, with the sugar hydrolases comprising an estimated 20--25% of the membrane protein. The immunoreactivity of membrane-bound sucrase-isomaltase, maltase, and lactase was investigated with antisera demostrating specific reactivity to each, when tested in the presence of other membrane extractives. The membrane-bound enzymes were found in each case to combine with antibody in amounts equivalent to that required to effect precipitation of comparable units of the free enzymes from solution. Preloading membrane vesicles with antibodies to any two of the enzymes did not affect either the immunoreactivity or extractability (by papain or Triton X-100) of the third. The antibody-binding studies indicated an arrangement of these enzymes independent of each other on the membrane surface, in a manner allowing each to maintain a high degree of molecular freedom.


Urethritis attributable to Neisseria meningitidis, group X: a case report. Neisseria meningitidis, Group X, was recovered from the anterior urethra of a 27-year-old male patient with urethritis. This isolate represents the first reported recovery of this organism from this anatomic site in association with this disease. He was treated with probenicid by mouth, followed by 4.8 million units of aqueous procaine penicillin G injected intramuscularly and became asymptomatic in three days. His recovery strongly indicates that the meningococcus was the etiologic agent of the urethritis since the condition promptly cleared with penicillin therapy.


Restriction cleavage map of SP01 DNA: general location of early, middle, and late genes. A detailed restriction endonuclease map for the genome of Bacillus subtilis phage SP01 is presented. Sites of cleavage for the restriction enzymes BglII, EcoRI, HaeIII, and SalI were determined. This physical map showed that SP01 DNA was 140 kilobases in length and contained a repeated sequence of 12.4 kilobases at its termini. Combined with previously published information, we were also able to identify the general locations of genes expressed at early, middle, or late times in the phage lytic cycle. In particular, early genes were largely clustered in the terminal repeats, whereas a major cluster of late genes was located in the left-central portion of the genome.


Brucella abortus in coyotes. I. A serologic and bacteriologic survey in eastern Texas. Prevalence of Brucella abortus serum antibodies in coyotes from east central Texas was determined by the buffered Brucella antigen (card test), rivanol, standard agglutination tube, and cold complement fixation tube tests. Eighteen percent (9 of 51) of the coyotes were positive serologically. B. abortus biotype 1 was isolated from various tissues from 7 of 43 coyotes by bacteriologic culture. Congenital transmission was found.


Bacteriophage T4-coded dihydrofolate reductase: synthesis, turnover, and location of the virion protein. Dihydrofolate reductase plays a dual role in bacteriophage T4, first, as an enzyme of thymidylate metabolism, and second, as a protein component of the tail baseplate. Antibody to the purified enzyme has been used to study its synthesis and intracellular turnover. The antibody specifically precipitates one protein from T4D-infected cell extracts. This has been identified as dihydrofolate reductase, although the polypeptide molecular weight (22,000) is lower than that earlier determined for this enzyme. The protein comigrates on gels with pY, a genetically undefined protein component of the baseplate. However, it is not pY, for pY is synthesized late in infection, whereas virtually no dihydrofolate reductase synthesis occurs later than 10 min after infection at 37 degrees C. Dihydrofolate reductase, once formed, is neither degraded nor converted to proteins of higher or lower molecular weight. Thus, it is probably incorporated into virions at the same molecular weight as that of the soluble enzyme. 125I-radiolabeled antibody binds to the wedge substructure of the baseplate, and this binding is blocked by preincubation with purified T4 dihydrofolate reductase. Thus, the enzyme protein seems to be a component of the wedge.


Inhibition of mammary tumorigenesis in carcinogen-treated Lewis rats by suppression of prolactin secretion. One hundred sixty-eight female Lewis rats were treated intragastrically with 10 mg 7,12-dimethylbenz[a]anthracene at 56 and 63 days of age. Pituitary prolactin secretion was suppressed in one-half of these rats by daily sc administrations of 2-bromoergocryptine mesylate (CB-154; 0.4 mg/100 g body wt) from 29 to 90 days of age (series 1) and from 90 to 140 days of age (series 2). Treatment with CB-154 was initiated prior to the onset of palpable mammary carcinomas. Control rats were given injections of saline. Inguinal mammary glands were excised from 10 control and 10 CB-154-treated rats at the cessation of saline and CB-154 treatments and examined for hyperplastic nodules (HN). The remaining rats were palpated weekly for mammary carcinomas (MC) and killed at 200 days of age. Mean number of HN per rat, mean number of MC per rat, and percent of rats with MC were, respectively: series 1--controls, 0.6, 1.5, and 68; CB-154 treatment, 0.5, 1.1, and 62; series 2--controls, 10.4, 2.0, and 94; CB-154 treatment, 5.1, 1.1, and 56. The number of HN and MC was only slightly reduced in rats when prolactin was suppressed during carcinogen treatment (series 1) but markedly reduced when prolactin was suppressed after carcinogen treatment (series 2). These results provide evidence that prolactin is involved in the early development of mammary dysplasias in the carcinogen-treated female Lewis rat.


[Treatment of chronic coronary insufficiency with Irrigor]. The results of treatment of 103 patients suffering from ischemic heart disease with irrigor for 7--25 days are discussed. The drug was given in a daily dose of 30 to 150 mg in tablets, intramuscularly, and intravenously. Ergometry, study of the values of central hemodynamics and lipid metabolism, and recording of the rheoencephalo-, rheovaso-, and electroencephalogram were conducted before and after the treatment. The efficacy of irrigor was appraised in treatment applied in courses, in a single intravenous infusion, and under conditions of the double blind method. A relatively stable clinical effect was noted on the 9th--12th day. A good effect was produced in 49.3% of patients. The efficacy of irrigor was the highest in patients with grade I and II chronic coronary insufficiency, with no history of circulatory insufficiency or myocardial infarction, and in intramuscular and intravenous administration. Treatment with irrigor improved the indices of central hemodynamics and cerebral and peripheral blood flow. No side effects of the treatment were noted. The study showed irrigor to be an effective agent in the treatment of ischemic heart disease.


Pregnancy diagnosis in owl monkeys (Aotus trivirgatus): evaluation of the hemagglutination inhibition test for urinary chorionic gonadotropin. The Subhuman Primate Pregnancy Test was evaluated as a means of detecting urinary chorionic gonadotropin to aid in pregnancy diagnosis in owl monkeys. Using radioimmunoassay, the excretion pattern of chorionic gonadotropin from pregnant owl monekys was delineated, the hormone being detected from 16 weeks prepartum until birth. By comparison, the pregnancy test kit detected chorionic gonadotropin between the fourteenth week prepartum and the last week of gestation with 94% accuracy. In a 2-year study using a simplified urine collection technique, the Subhuman Primate Pregnancy Test was shown to be a valuable procedure for diagnosing pregnancy and detecting spontaneous abortions in owl monkeys.


Cholinesterase activity in Japanese quail dusted with carbaryl. Japanese quail (Coturnix coturnix japonica) were dusted with 5% carbaryl to determine if this topical treatment would alter plasma and brain cholinesterase activities. Within 6 hours after dusting, plasma cholinesterase activity was depressed compared with controls, the depression averaging 20% for females and 27% for males. By 24 hours the cholinesterase activity of females had returned to normal, but the cholinesterase activity of males remained depressed. Brain cholinesterase activity was not affected by the treatment, and there were no overt toxic signs.


Clinical utilization of mechanical sutures for intestinal anastomosis in Macaca mulatta. Using three mechanical stapling instruments that employ stainless steel wire staples, a Mann-Bollman fistula was created on the terminal ileum in 27 rhesus monkeys (Macaca mulatta). Two functional end-to-end anastomoses were accomplished and an ileostium constructed. Staples of various sizes were used; it was found that pediatric-sized staples were necessary for the intestinal tissue of these primates. Six months after surgery, microscopic examination revealed encapsulation of the staples by granulomatous inflammatory tissue. When staples of the proper size were selected, mechanical suturing provided rapid and consistent anastomoses, and wound closures had less edema, inflammation and necrosis than with conventional suturing methods.


Root canal procedure for disarming nonhuman primates. Nonhuman primates were disarmed by shortening their canine teeth. These teeth were cut off near the gingival level, the entire pulpal tissue removed and the canal filled with a formulated paste which was radiopaque, adhesive and germicidal. The teeth were sealed with a commercial alloy. This endodontic procedure was a quick, practical one-step method for permanently disarming nonhuman primates.


Multiple cannulation of the primate superficial lateral coccygeal vein. The superficial lateral coccygeal veins of Macaca fascicularis were exposed surgically and cannulated with polyethylene tubing. The cannula was used for administering continuous infusions or obtaining multiple blood samples, and it was removed 12--18 hours after insertion.


Bilateral inguinal hernia in a pig-tailed monkey (Macaca nemestrina). A pig-tailed monkey developed bilateral inguinal hernias following escape from its cage and subsequent recapture. The hernias were surgically repaired and the monkey recovered.


Deranged isoleucine metabolism during ketotic attacks in patients with methylmalonic acidaemia. Two patients with methylmalonic acidaemia due to methylmalonyl-CoA mutase deficiency were studied for several years. Both exhibited at least two attacks of severe ketoacidosis, during which they excreted, in addition to methylmalonic acid, a number of abnormal compounds: 3-hydroxypropionic acid, 2-methyl-3-hydroxybutyric, 3-hydroxy-n-valeric acid, 3-oxo-n-valeric acid, 2-methyl-3-oxobutyric acid, citraconic acid and N-tiglyglycine. These compounds represent partly intermediary metabolites from the isoleucine degradation pathway and partly secondary metabolites of propionyl-CoA and tiglyl-CoA.


Diagnosis of Gaucher's disease in cultured skin fibroblasts and leucocytes. Washing skin fibroblasts or leucocytes in 0.25 mol/l sucrose increases the activity of beta-glucosidase at acid pH. This effect is primarily due to removal of low levels of sodium chloride, which inhibit acid beta-glucosidase. A secondary factor for skin fibroblasts in the removal of residual phosphate buffer pH 7.3 used to wash the cells following trypsinization. As the beta-glucosidase activity of water-lysed leucocytes is higher at acid pH than that of a saline suspension of leucocytes, the former are better for the diagnosis of Gaucher's disease. However, more reliable results still may be obtained by assay of this enzyme in cultured skin fibroblasts.


The use of natural and artifical substrates in the prenatal diagnosis of Krabbe's disease. Krabbe's disease was diagnosed prenatally using cultured amniotic fluid cells and the diagnosis confirmed using fetal brain, liver and cultured fetal skin fibroblasts. The enzyme defect was demonstrated by assay of galactocerebrosidase and lactocerebrosidase I, and by hydrolysis of the chromogenic analogue, 2-hexadecanoylamino-4-nitrophenyl-beta-D-galactopyranoside. The relative merits of the three diagnostic methods are discussed.


Non-ketotic hyperglycinaemia in a family with an unusual phenotype. A 10-year-old girl, one of three affected sisters, with non-ketotic hyperglycinaemia is described. In contrast to other reported cases, the course of the disorder was comparatively mild in this family. The only clinical signs were mental retardation and abnormalities in the EEG; blood glycine levels were 2-3 times normal. In the propositus, the formation of 14CO2 from glycine-1-14C and of FH414CH2OH from glycine-2-14C were impaired, shown by the decreased 14CO2 content of expired air and diminished labelling of carbon 3 of serine. However, the biochemical defect was no less than that seen in patients with much more severe clinical effects.


A sensitive semi-automated kinetic assay of alpha-D-glucosidase for the prenatal diagnosis of type 2 glycogenosis (Pompe's disease). Prenatal diagnosis of type 2 glycogenosis (Pompe's disease) has been done on cultured amniotic fluid cells, using a semi-automated fluorimetric kinetic assay for alpha-D-glucosidase with 4-methylumbelliferyl-alpha-D-glucoside as substrate. The activity of the enzyme was related to that of beta-D-galactosidase, and found to be absent in cells from an affected fetus. The diagnosis was confirmed in fetal liver, where the same assay was used to show absence of alpha-D-glucosidase activity with normal beta-D-galactosidase activity, and where increased glycogen deposition was demonstrated histologically. This type of assay is generally applicable to lysosomal enzymes, and to other fluorigenic enzyme reactions.


Familial hyperlysinaemia due to L-lysine alpha-ketoglutarate reductase deficiency: results of attempted treatment. A mentally retarded male infant with persistent hyperlysinaemia due to L-lysine alpha-ketoglutarate reductase deficiency is described. The effect of dietary restriction of lysine on his mental and behavioural development was examined. By restricting daily dietary lysine to 5.5 mg/kg body weight the fasting serum lysine became normal. Urinary lysine also became normal and the secondary metabolites homocitrulline, homoarginine, N alpha-acetyllysine and N epsilon-acetyllysine were no longer detected. After control of serum lysine for 2.5 y it was felt that the patient's social behaviour, but not his mental development, had improved somewhat.


Aspartylglycosaminuria in Northern Norway in eight patients: clinical heterogeneity and variations with the diet. Urinary excretion of aspartylglycosamines was investigated in eight patients by semiquantitative thin-layer chromatography, and bound glycosamines by a quantitative photometric method (Elson-Morgan reaction). Each patient showed a fairly constant level, relative to the creatinine, of aspartylglycosamines in urine. The least retarded patient, aged 31, excreted about 350 mg/g creatinine, one-third of that found in two severely retarded young patients, aged 4 and 7 years (1400 and 940 mg/g creatinine, respectively). Three days on a low-protein diet did not change the aspartylglycosamine excretion in the patient showing the highest excretion rate.


Diaphragmatic pedicle flap for treatment of Boerhaave's syndrome. A case of spontaneous perforation of the esophagus is described, which was managed by thoracotomy, primary closure, and reinforcement of the suture line with as diaphragmatic pedicle flap. The advantages of this technique are discussed.


Studies on the conjugation of horseradish peroxidase to immunoglobulin G via glutaraldehyde. Horseradish peroxidase was conjugated to immunoglobulin G via glutaraldehyde by a two-step procedure using an increasing excess of peroxidase in the second step reaction. The yield of conjugated monomeric IgG and the amount of free IgG were analyzed by SDS-polyacrylamide electrophoresis and gel-filtration. The antigen binding capacity of the enzyme-antibody conjugates was evaluated by radial immunodiffusion. Conjugation of peroxidase to IgG with a 1:20 molar:molar excess of glutaraldehyde-activated peroxidase resulted in a high yield of conjugated IgG without any detectable amounts of polymers of IgG or residual free IgG. The antigen binding capacity of the conjugate varied between different antigen-antibody systems, but in general it was not significantly different from that of native IgG. The enzyme activity was reduced to 70% of the activity of native peroxidase.


Nutritional assessment of the hospitalized patient. While some symptoms of the malnourished state are obvious, others may be subtle. In either case, assessment is indicated to uncover any inconspicuous symptoms and secondly to quantitate those that are apparent. Such procedures are pertinent because immune competence and other organ systems related to survival are dependent on the adequately nourished state. It is also important to remember that certain vitamin and mineral deficiencies may accompany protein calorie malnutrition and thus their attention would be especially needed during the repair of this condition. The deficiencies seen most often include vitamins C, A, and D, as well as zinc, copper, and calcium. Furthermore, the refeeding process may also precipitate shortages of potassium, phosphate, and magnesium, unless adequate amounts of these nutrients are included in the diet. From this and earlier discussed situations, it is suggested that nutritional assessment should be the initial step in nutritional therapy. It should be approached with concern not only for what the collective assessment tools will reveal, but also with consideration for the patient's clinical problem, the current metabolic situation and the projected therapeutic regimen which will be formulated.


Hyperalimentation.

Hyperalimentation. As widespread use of parenteral nutrition enters the second decade, it is obvious that a great deal of progress has been made, both as to increasing safety and developing new solutions. There are few hard data in this area, not surprising in so young a field. Prospective studies hopefully will yield definitive results in areas in which such knowledge is lacking. More importantly, better understanding of various disease states will undoubtedly result in the modification of existing solutions to allow for better treatment.


Paranasal sinus aspergillosis.

Paranasal sinus aspergillosis. Four cases of aspergillosis of the paranasal sinuses seen recently at the North Carolina Baptist Hospital are reported. Paranasal sinus aspergillosis is prone to develop in patients living in the southeastern states because those states 1. have an agricultural economy and 2. have a hot, humid climate that causes a high incidence of nasal and sinus disorders. The increasing use of immunosuppressive and oncological drugs should further increase the incidence of aspergillosis. Diagnosis of aspergillosis of the paranasal sinuses requires a high index of suspicion which should be present particularly for a patient who has unilateral sinusitis that does not respond to routine therapy, or who has a localized sinus mass or bony erosion. Immunodiffusion tests are specific for aspergillosis. Treatment consists of surgical removal of the diseased mucosa and adequate aeration and drainage of the sinus. Culture of the specimen on Sabouraud's agar will confirm the diagnosis.


Hypopharyngeal diverticulum and the cricopharyngeus muscle: a posterior surgical approach. Hypopharyngeal diverticulum and dysphagia due to cricopharyngeus muscle dysfunction are a continuing challenge to the head and neck surgeon. The ability to completely transect the cricopharyngeus muscle is generally agreed to be the key to successful relief of symptoms, and--to an even greater extent--to prevention of recurrence. In the past, the most significant complications arising from such surgery have been recurrent laryngeal nerve injury and recurrent diverticulum or cricopharyngeus spasm. Both of these problems have been prevented by a modified surgical approach which takes advantage of the relatively bloodless retropharyngeal space. The logic of this posterior approach to the cricopharyngeus, from an anatomic standpoint, coupled with the good visualization and relatively bloodless field when approaching the muscle, suggests that it might be a suitable substitute for the more common lateral approach if continued long-term experience in larger number of cases does not lead to significant complications.


Respiration-coupled calcium transport by membrane vesicles from Azotobacter vinelandii. Membrane vesicles, isolated from osmotic lysates of Azotobacter vinelandii spheroplasts in Tris-acetate buffer, rapidly accumulate calcium in the presence of an oxidizable substrate. The addition of D-lactate to vesicles increases the rate of calcium uptake by 34-fold; L-malate, NADH, NADPH, and reduced phenazine methosulfate are nearly as effective as lactate. The intravesicular calcium pool which accumulates under these conditions is rapidly discharged by isotopic exchange or in the presence of respiratory inhibitors, uncouplers, or EGTA. The uptake rates for calcium follow Michaelis-Menten kinetics yielding a Km of 48 microM and a V max of 45 nmoles/min/mg membrane protein. Initial rates of EGTA-induced calcium efflux also follow saturation kinetics, giving a V max identical to that for calcium entry; but the Km for exodus is 14 mM, assuming that free calcium accumulates in vesicles. The difference in the affinity of calcium for the entry and exit processes observed during respiration is sufficient to account for the estimated 150-fold calcium concentration gradient achieved under steady-state conditions. The uptake system is specific for calcium as opposed to other cations, but zinc and lanthanum are effective competitors. Calcium uptake is blocked when electron is inhibited by exposure of vesicles to p-chlormercuriphenylsulfonate, hydroxyquinoline-N-oxide, or cyanide, or under anoxic conditions. Divalent cation ionophores (A23187 and X537A) and proton ionophores (CCP and gramicidin D) also block calcium transport effectively. The electrogenic potassium ionophore valinomycin has no effect on lactate-dependent calcium uptake in the presence of potassium; but ionophores which induce electroneutral exchange of protons for sodium or potassium (monensin and nigericin, respectively) did block calcium transport in the presence of the appropriate cation. The fluorescence intensity of quinacrine (an amine probe) in the presence of A. vinelandii membrane vesicles is reduced by 25% on addition of lactate; the quenching is blocked by CCP. This indicates that a pH gradient (inside acid) is developed across the vesicle membrane during lactate oxidation. These results indicate that these membrane preparations contain vesicles of inverted topology (with respect to the intact cell) and suggest that calcium transport occurs by means of electroneutral calcium/proton antiport.


Primary Ewing's sarcoma of the sternum: a case report. A 12 1/2-year-old female presented with Ewing's sarcoma of the manubrium sterni which extended into the anterior mediastinum. At presentation there was no evidence of metastatic disease. Her initial treatment consisted of radiation therapy and chemotherapy. The residual tumor was subsequently resected and the sternal defect was repaired with Marlex mesh. Postoperatively, she was maintained on chemotherapy consisting of BCNU, cyclophosphamide, and adriamycin. The adriamycin was discontinued after she developed sterile fibrinous pericarditis. She remains free of her disease two years after diagnosis. Although extremely rare, Ewing's does occur in the sternum and this area is amenable to wide local resection without severe functional disability.


Successful therapy of crystalcryoglobulinemia: a case report. Successful therapy for a case of multiple myeloma with a spontaneously crystallizing cryoglobulin of the IgG2-kappa light chain variety was achieved, using both continuous-flow cell centrifugation plasmapheresis to rapidly lower the M component and combination chemotherapy with phenylalanine mustard, prednisone, procarbazine, and vincristine to control the myeloma process. This resulted in resolution of incapacitating large and small necrotic cutaneous ulcerations of the extremities. Physicochemical studies of the crystalcryoprotein demonstrated that cryoprecipitation was rapid and accompanied by the formation of needle-shaped crystals, yet was completely reversible at 37 degrees C. Cryocrit determinations varied depending upon relative centrifugal forces and temperature and did not always relate linearly to the amount of abnormal protein, thus making these alone unreliable in assessing response to therapy.


[Lipoamino acids in the composition of actinomycete polar lipids]. The capacity for substitution of phospholipids, in particular phosphatidyl ethanolamine, by a phosphorus-lacking lipoamino acid was studied in different groups of actinomycetes. In the conditions of phosphorus deficiency, most cultures were found to be capable of synthesizing a phosphorus-lacking nitrogen-containing lipoamino acid. Its characteristics (Rf) in TLC are similar to those of ornithinolipid. This fact confirms, for the first time and for different groups of actinomycetes, the hypothesis concerning interchangeability of membranous lipids. In the case of Actinomyces olivaceus, phosphatidyl ethanolamine and ornithine containing a phosphorus-lacking lipid can be regarded, this being corroborated by comparative data about their quantitative content. The detection of the phosphorus-lacking nitrogen-containing lipid in the composition of the polar fraction in different actinomycetes in the conditions of phosphorus deficiency in the medium demonstrates the possibility of adaptive changes in the composition of membranous lipids in microorganisms depending on their habitat.


[Growth and biomass accumulation by several strains of Pseudomonas on nutrient with ethanol]. The capacity to grow in a mineral medium with ethanol as a sole carbon source was studied in 147 Pseudomonas strains. All of the strains can grow in this medium. The biomass accumulation in the mineral medium with ethanol (1%) and maize extract (0.2%) is 2.5--5.0 times higher than without the latter. Strains which accumulate up to 3.5 g of dry biomass per litre (containing 62--66% of protein and 5.3% of nucleic acids) in the mineral medium with ethanol (1%) and maize extract (0.2%) have been selected. The quantitative content of essential amino acids satisfies the requirements while that of lysine, threonine, valine and leucine is twice as high.


[Diauxotrophic properties of microorganisms assimilating C2--C4 hydrocarbons]. Diauxotrophic properties of bacteria assimilating gaseous hydrocarbons C2--C4  and other complex organic substances but not methane were studied. If the medium contained two substrates (hydrocarbon+carbohydrate), the non-growing cells of the strains did not display diauxotrophic properties. In the phase of exponential growth, oxygen-containing carbon sources and then gaseous hydrocarbons were assimilated, i.e. diauxia was observed. If a microbial association containing an obligate methylotroph and a facultative gas-assimilating culture was grown on a medium with a natural gas, the latter culture assimilated carbon-containing metabolites of the methylotroph and then, when their concentration decreased, gaseous hydrocarbons. The order in which complex organic substances (exometabolites of methylotrophs) and hydrocarbons C2--C4 were assimilated was determined by their concentration in the medium. In the course of growth of such a microbial association, the inhibiting effect of metabolites of methylotrophs on their growth decreased as well as the loss of methane being transformed by methylotrophs into exometabolites, and hydrocarbons C2--C4 were utilized.


[Morphologic heterogeneity under different conditions of living and destruction in Anabaena variabilis cyanobacteria]. The proportion between various morphological forms of the cyanobacterium Anabaena variabilis was studied under different conditions of its growth and destruction. When the cells lost viability at the stationary phase of cultural growth under the optimal conditions of illumination (1500--2000 lx), in the dark or when nitrogen was deficient, the filaments became shorter (4 to 2.2 cells per filament on the average), the cells larger, 4.5x6 microns). Under the conditions of nitrogen deficiency, the content of phycocyanin sharply decreased as well as that of chlorophyll. "Aging" under the conditions of optimal illumination was accompanied with "weighting" of the cells at the prolonged stationary phase and a gradual decrease in the content of phycocyanin and chlorophyll. When the cells were exhausted in the dark, the content of protein, RNA and phycocyanin decreased while that of chlorphyll hardly changed for a considerably long period of time. The most versatile morphological heterogeneity, when the cells were rapidly destroyed under the conditions of high light intensity, was as follows: both larger and more spherical as well as smaller and longer than in the control cells was found and catenuate cells were detected. The colour of the culture was light-brown or blue. When grown in a medium containing aspartic acid, the culture consisted of long filaments (ca. 18 cells per fragment instead of 4 cells in the control).


Prostaglandin synthetase inhibitors and food intolerance. 12 patients with adverse reactions to foods are discussed, including 1 with biopsy-proven ulcerative proctitis. 10 had predominantly gastrointestinal symptoms (diarrhoea, abdominal pain, vomiting) and two had rhinitis and headache. Skin tests (in all but two) and radioallergosorbent tests (RAST) in some patients were negative. All of the patients were challenged in hospital with the offending food, either alone or preceded by a prostaglandin synthetase-inhibiting drug (aspirin, indomethacin or Ibuprofen). In 11 of the 12 patients this premedication prevented both the gastrointestinal and the more remote symptoms. Blood and stool prostaglandin measurements (PGE2 and PGF2 alpha) showed changes which correlated with clinical symptoms and did not occur if one of the inhibiting drugs had been given prior to challenge.


Xenobiotica-metabolizing enzymes in Drosophila melanogaster: activities of epoxide hydratase and glutathione S-transferase compared with similar activities in rat liver. Activities of epoxide hydratase and glutathione (GSH) S-transferase were investigated in subcellular fractions of Drosophila melanogaster, and these activities were compared with analogous enzymic activities in extracts from rat liver. Microsomes of Drosophila were active in the hydratation of styrene oxide catalyzed by epoxide hydratase. The post-microsomal supernatant of Drosophila catalyzed the conjugation of GSH with 1-chloro-2,4-dinitrobenzene. However, GSH S-transferase activity with styrene oxide as the electrophilic substrate was not measurable. The respective specific activities of epoxide hydratase (per mg microsomal protein) and GSH S-transferase (per mg cytosolic protein) were factors of 5- and 10-fold lower than the corresponding activities in rat liver. However, when expressed per gram body weight, activities of both epoxide hydratase and GSH S-transferase were 3 times higher for Drosophila enzymes. The apparent Km values for the two Drosophila enzymes were higher, whereas the apparent Km values were lower, than the values found for the rat-liver enzymes. Among 3 different Drosophila strains (a wild-type, a white eye-color carrying mutant strain and a DDT-resistant strain), preliminary experiments showed no differences as far as these two enzymic activities were concerned. It is concluded that the results obtained in genetic toxicology testing with Drosophila are probably relevant to effects to be expected in mammalian systems with compounds requiring metabolic processes involving the enzymes investigated here.


Is multiple sulphatase deficiency due to defective regulation of sulphohydrolase expression? Multiple sulphatase deficiency disease is an unusual autosomal recessive disorder characterised biochemically by a deficiency of several sulphohydrolase activities. The laboratory diagnosis of this combined neurological connective tissue disorder is made on the basis of decreased activities of the lysosomal enzymes, arylsulphatase A and arylsulphatase B and the microsomal enzyme, arylsulphatase C. The primary defect in this multi-enzyme deficiency has not been identified. Using immunological techniques to characterise further the residual activities of arylsulphatases A and B in the multiple sulphatase deficiency disease, we have examined the levels of cross-reaching material (CRM) to arylsulphatases A and B in cultured skin fibroblasts from controls and patients with multiple sulphatase deficiency, metachromatic leukodystrophy (deficiency of only arylsulphatase A activity) and Maroteaux-Lamy syndrome (deficiency of only arylsulphatase B activity). We report here results indicating that arylsulphatases A and B in multiple sulphatase deficiency are reduced in their levels of CRM while retaining a normal activity/CRM ratio. Because the two enzymes are apparently structurally unrelated, these data are consistent with the possibility that their combined deficiencies in this disorder may result from a defect in the coordinated expression of sulphohydrolases.


A modified RNA polymerase transcribes a cloned gene under sporulation control in Bacillus subtilis. A modified form of RNA polymerase from Bacillus subtilis selectively transcribes a cloned gene under early sporulation control. This RNA polymerase lacks sigma factor but contains a newly identified subunit of molecular weight 37,000, termed P37. P37 could be a regulatory protein that controls, at least in part, an early stage of spore development.


A region of the Drosophila genome necessary for CNS development. Mutations in genes involved in essential aspects of central nervous system development in Drosophila melanogaster are expected to be lethal. Thus, when searching for neurogenic mutants attention should be focused on embryonic lethal point mutants, for many of these might affect neural development. However, this approach can be very time consuming, for the location of neurogenic genes is unknown. A more convenient approach, which allows a faster screening of the genome, is to use relatively small chromosome deletions to determine whether the lack of a definite part of the genome affects neurogenesis. Once any region producing an interesting neural phenotype is found, it can be further analysed by the use of smaller deletions or point lethal mutants mapping within it, until the gene(s) responsible can be more precisely localised. We report here on a region of the Drosophila genome which has been found necessary for normal neurogenesis.


Cis-active control of mouse beta-galactosidase biosynthesis by a systemic regulatory locus. Several higher organisms have been reported in which enzyme levels are determined genetically by sites located in close proximity to the corresponding structural genes. In several cases, these sites have been shown to act by controlling the rates of enzyme synthesis. Cis compared with trans action has been tested for those proximate regulatory sites controlling enzymes for which appropriate structural variants exist. The rate of synthesis of beta-galactosidase in mouse tissues is under the control of a regulatory locus; Bgl-s, that is tightly linked to the enzyme structural gene; we have tested the cis/trans nature of Bgl-s action by analysis of the electrophoretic mobility of the enzyme from animals heterozygous for the appropriate regulatory and structural alleles. Our results indicate that Bgl-s acts cis, controlling the expression of the structural gene located on the same chromosome.


The dependence of coronary collateral blood flow on regional vascular resistances. Pharmacological studies with glyceryl trinitrate, adenosine and verapamil. A method was applied in anesthetized dogs enabling the measurement of regional resistances up to and behind the start of collaterals and the collateral resistance. The studies show that peripheral coronary pressure, i.e. perfusion pressure of the collaterals, can change when the ratio of pre- and post-collateral resistance alters. Drugs can influence collateral blood flow not only by directly effecting the collaterals but also by altering collateral perfusion pressure. Glyceryl trinitrate given in minor doses improved collateral blood flow by directly dilating the collaterals and also by increasing collateral perfusion pressure. Higher doses did not improve collateral flow due to a decrease of collateral perfusion pressure. A steal-phenomenon occurred in some cases. Adenosine and verapamil had no direct influence on the collateral resistance. Verapamil given in small doses increased perfusion pressure slightly but not enough to improve collateral blood flow. High doses of verapamil, like low doses of adenosine, had no significant influence on collateral perfusion pressure and collateral blood flow. Adenosine given in high dosage led to a diminution of collateral flow by decreasing collateral perfusion pressure, i.e. a steal-phenomenon.


Ultrastructure of somatotrophs of rats with median eminence lesions: studies in basal conditions and after thyrotropin-releasing hormone stimulation. Anterior pituitaries from female Sprague-Dawley rats (160-200 g b.w) bearing electrolytic lesions of the median eminence were qualitatively and quantitatively investigated by electron microscopy and the findings compared with those from intact animals or sham-operated controls. This study was performed in basal conditions and after stimulation with thyrotropin-releasing hormone. Animals with lesions showed a reduction of both the number of somatotrophic cells and growth hormone granules. After injection of thyrotropin-releasing hormone exocytosis of growth hormone granules from somatotrophs was frequent in rats with lesions of the median eminence but rare in sham-operated animals. The differences were statistically significant.


Plasma concentrations of phensuximide, methsuximide, and their metabolites in relation to clinical efficacy. The clinical efficacy of phensuximide and methsuximide was studied in relation to plasma concentrations of these compounds and their desmethyl metabolites. Single- and chronic-dose studies of each drug were carried out in five patients with intractable seizures. Patients were evaluated before and during treatment by 6-hour simultaneous video and telemetered electroencephalographic recordings to characterize the seizure type and by daily determinations of plasma antiepileptic drug concentrations. Phensuximide had a mean half-life of 7.8 hours and accumulated to an average fasting level of only 5.7 micrograms per milliliter. Desmethylphensuximide averaged only 1.7 micrograms per milliliter with a similar half-life. Methsuximide had an even shorter half-life, averaging 1.4 hours, but its desmethyl metabolite had a mean half-life of 38 hours and therefore accumulated to levels in excess of 40 micrograms per milliliter. The addition of phensuximide to their regimens benefited none of the patients, but two had an excellent response to methsuximide. The failure of phensuximide and its desmethyl metabolite to accumulate to reasonable levels is the likely explanation for the relatively weak antiepileptic effect of phensuximide as compared with methsuximide.


Mineralization of the stylohyoid-stylomandibular ligament complex. A radiographic incidence study. In this Veterans Administration-based study, 1,771 oral panoramic radiographs were reviewed to detect abnormalities in the styloid process, stylohyoid-stylomandibular ligament complex. Three hundred twenty-three (18.2 percent) of the radiographs revealed mineralization in these areas which varied from an apparent elongation of the styloid process to extensive involvement of the entire ligament complex with segmentation, nodularity, and the development of jointlike structures. Mineralization was unilateral in 12.4 percent of patients and was observed more commonly in patients 50 to 59 years of age. No relationship could be established between the extent of mineralization and increasing patient age. This study suggests that mineralization of the stylohyoid complex is not uncommon and probably only rarely causes symptoms.


Pulpal response to bacteria in the dog.

Pulpal response to bacteria in the dog. Twenty-six teeth from 8-month-old beagle dogs were used to evaluate the pulpal response to bacteria in bacterially contaminated minimally exposed and unexposed pulp in the dog. The effects were evaluated by means of serial radiographs, consecutive vital dye injections, and histologic sections. Periapical lesions developed in all of the eight teeth which were bacterially infected and exposed. Out of ten teeth which were bacterially infected and had cavities without pulp exposure, only two teeth in the acute group demonstrated inflammatory changes and loss of odontoblastic function directly beneath the infected dentinal tubules. Within the limits of the experimental design, this study demonstrated the pulpal responses of the dog with respect to bacteria.


Soil stabilization by a prokaryotic desert crust: implications for Precambrian land biota. A cyanophyte dominated mat, desert crust, forms the ground cover in areas measuring hundreds of square meters in Utah and smaller patches in Colorado. The algal mat shows stromatolitic features such as sediment trapping and accretion, a convoluted surface, and polygonal cracking. Sand and clay particles are immobilized by a dense network of filaments of the two dominating cyanophyte species, Microcoleus vaginatus and M. chthonoplastes, which secrete sheaths to which particles adhere. These microorganisms can tolerate long periods of desiccation and are capable of instant reactivation and migration following wetting. Migration occurs in two events: 1. immediately following wetting of dry mat, trichomes are mechanically expelled from the sheath as it swells during rehydration, and 2. subsequently, trichomes begin a self-propelled gliding motility which is accompanied by further production of sheath. The maximum distance traveled on solid agar by trichomes of Microcoleus vaginatus during a 12 hour period of light was 4.8 cm. This corresponds to approximately 500 times the length of the fastest trichome, and provides a measure of the potential for spreading of the mat in nature via the motility of the trichomes. Dehydration resistence of the sheath modifies the extracellular environment of the trichomes and enables their transition to dormancy. Following prolonged wetting and evaporative drying of the mat in the laboratory, a smooth wafer-like crust is formed by the sheaths of Microcleus trichomes that have migrated to the surface. Calcium carbonate precipitates among the algal filaments under experimental conditions, indicating a potential for mat lithification and fossilization in the form of a caliche crust. It is suggested that limestones containing tubular microfossils may, in part, be of such an origin. The formation of mature Precambrian soils may be attributable to soil accretion, stabilization, and biogenic modification by blue-green algal land mats similar to desert crust.


Aldose reductase and sorbitol dehydrogenase in the muscle of Ascaris suum (Nematoda). The muscle of Ascaris suum contains sorbitol dehydrogenase [EC 1.1.1.14] and the apparent Km values indicate that the enzyme favours fructose formation. Aldose reductase [EC 1.1.1.21] is also present and like the mammalian enzyme it has a very high apparent Km for glucose. Possible functions of the sorbitol pathway in Ascaris muscle are discussed.


Carrier detection of pyruvate carboxylase deficiency in fibroblasts and lymphocytes. Pyruvate carboxylase (E.C. 6.4.1.1) activity was determined in the circulating peripheral lymphocytes and cultured skin fibroblasts from the family of a patient with hepatic, cerebral, renal cortical, leukocyte, and fibroblast pyruvate carboxylase deficiency (PC Portland deficiency). Lymphocyte activities were: mother, 33--39%; father, 11--29%; brother, 82--103%; and sister, 38--48% of the lowest normal. Fibroblasts from the patient's mother and father had 42 and 34%, respectively, of the activity of the lowest normal. These data demonstrate that the disease is inherited in an autosomal recessive manner and that lymphocytes and fibroblasts can be used to detect carriers. Neither pyruvate carboxylase nor mitochondrial PEPCK activity in lymphocytes was increased by a 21-hr fast.


Inhibitory effect of cystic fibrosis serum on pseudomonas phagocytosis by rabbit and human alveolar macrophages. This report presents experimental observations indicating the presence of an inhibitory activity in cystic fibrosis (CF) serum which impairs phagocytosis of Pseudomonas aeruginosa by rabbit as well as human alveolar macrophages. Of the 49 patient serum samples studied, 40 consistently showed greater than or equal to 60% inhibition, 3 showed no inhibition and 6 were in the range of 20-60% inhibition of Pseudomonas phagocytosis. In parallel studies, the phagocytosis of S. aureus and S. marcescens was found not to be inhibited by CF serum. Mixing of CF serum with normal serum could not overcome the inhibitory effect, indicating the presence of an inhibitory factor rather than the lack of a necessary component. The inhibitory activity is not lost upon exposure of serum to glass, upon freezing the serum once, or upon heating at 56 degrees C for 30 minutes.


Invasive mycotic infections in the immunocompromised child. Oncologic therapy creates a highly vulnerable group of children with little or no natural immunity. In addition to the problems of coagulopathies, these children are prone to unusual infectious diseases. One group of saprophytic infections, the mycotic organisms, represents stubborn and often lethal diseases challenging present diagnostic and therapeutic skills. This is a case report of invasive Aspergillus organisms in the nose and premaxilla in a 9-year-old child suffering from acute monocytic leukemia.


Catastrophic necrosis of the neck.

Catastrophic necrosis of the neck. This case report illuminates the high possibility of catastrophic complications in the lateral neck, the vagaries associated with the preoperative therapeutic information, the technical effort to gain maximum surgical security, and the overwhelming complications associated with chemotherapy, irradiation, and surgical intervention.


Immunological purification and partial characterization of variant-specific surface antigen messenger RNA of Trypanosoma brucei brucei. Polyadenylated RNA isolated from total polyribosomes of two variable antigen types (VATs) of T. brucei brucei were shown to program the synthesis, in mRNA-dependant reticulocyte lysates, of a wide variety of polypeptides. After immunoprecipitation of these cell-free products with an homologous antiserum raised against purified variant-specific surface antigen (VSSA), a major electrophoretic band was apparent on fluorography. It was confirmed that this band corresponds to the variable antigen since only an excess of purified homologous antigen will provoke competition. The apparent molecular weight of the in vitro synthesized antigen is about 63,000 daltons. The VSSA mRNA has been found in membrane-bound polyribosomes and a 15 fold immunological purification of this mRNA has been obtained, using partially purified anti-VSSA IgG in conjunction with inactivated Staphylococcus aureus.


Analysis of RNA secondary structure by photochemical reversal of psoralen crosslinks. Aminomethyltrioxsalen (AMT), a psoralen, is known to cause interstrand crosslinks in double stranded nucleic acids. We have demonstrated the photochemical reversal of this reaction, and have used this result to develop a method for identification of specific sequences which are adjacent because of RNA secondary structure formation. E. coli 5S rRNA is used as a model system. We isolated and characterized a product that is derived from the stem region of 5S RNA.


The nucleotide sequence of tRNA tyrosine from the fission yeast Schizosaccharomyces pombe. The sequence of tRNA tyrosine from the fission yeast Schizosaccharomyces pombe is pCUCCUGAUm1 GGUG psi AGDDGGDDAUCACACor (psi) CCGGUG psi Ai6 AACCGGUUGm7 GUm5C GCUAGT psi CGm1 AUUCUGGUCAGGAGACCAOH. This sequence differs in 30 nucleotides from the tRNA-Tyr seqence of the budding yeast Saccharomyces cerevisiae. It has a unique anticodon stem of only four GC base pairs. The normal fifth pair position of nucleotide 28-44 is occupied by a C-U and in 20% of the tRNA-Tyr molecules it is psi-U. This unusual feature and its implications are considered in the discussion.


Series of 4.5S RNAs associated with poly(A)-containing RNAs of rodent cells. Uninfected mouse kidney cells and mouse leukemia cells L1210 in cluture contained a series of 4.5S RNAs which was structurally identical to the series of 4.5S RNAs associated with genomic RNAs of murine retroviruses and poly(A)-containing RNAs from virus infected cells. Normal rat kidney cells and baby hamster kidney cells in culture also contained a series of 4.5S RNAs. The strucutre of the 4.5S RNAs from mouse, rat and hamster cells were very similar, but not identical. These 4.5S RNAs were not found in cultured cells of other vertebrates, such as human, monkey, cat, mink, rabbit and chicken cells.


Studies on the specificity of preribosomal RNA transcription in nucleoli after selective deproteinization. Fidelity of preribosomal RNA transcription in vitro was studied after selective deproteinization of nucleoli using either sequential salt extraction or sodium deoxycholate treatment. Homochromatography fingerprinting and identification of marker oligonucleotides from a T1 ribonuclease digest of the transcripts were used to evaluate the RNA products. These studies indicated that: (1) nucleoli retained their endogenous RNA polymerase I activity and the specificity of transcription up to 0.6 M NaCl extraction; (2) exogenous RNA polymerase I transcribed nucleolar chromatin only after 1.0 M NaCl extraction and the transcription pattern, like that of totally deproteinized DNA, was completely random; (3) extraction of nucleoli with deoxycholate resulted in a DNP complex in which the endogenous RNA polymerase I transcribed pre-rRNA specifically; however, it also initiated random transcription, producing a "mixed" fingerprint pattern on the homochromatogram. The random transcription was selectively inhibited either by deoxycholate or rifampicin AF/013. These studies indicate that the selectivity of pre-rRNA transcription is due both to the endogenous RNA polymerase I molecules that were involved in transcription in vivo and are tightly bound to the template and to factors in intact nucleoli which prevent random transcription by the released RNA polymerase I molecules.


Separation of satellite DNA chromatin and main band DNA chromatin from mouse brain. Using restriction endonucleases which preferentially digest mouse main band DNA and leave satellite DNA intact, we have isolated highly purified chromatin fractions containing only mouse satellite or main band DNA. Following the digestion of mouse brain nuclei with EndoR Alu I, main band DNA chromatin is selectively extracted with 10mM Tris, 10mM EDTA. Satellite DNA chromatin is subsequently extracted from the nuclear pellet with Tris-3M urea and further purified on sucrose gradients. Chromatin extracted from digested nuclei with Tris-EDTA contains only main band DNA and has a molecular weight lower than 2 x 10(6). Chromatin fractions obtained from the lower regions of sucrose gradients of the Tris-Urea extracts contain 40--95% satellite DNA and have a molecular weight of 6 to 8 x 10(6). Both the satellite DNA and main band DNA chromatins contain all five histones and have a protein to DNA ratio of 1.3 to 1.


The nucleotide sequence of human tRNAGly (anticodon GCC). The sequence of tRNAGCCGly from human placenta was determined by recently developed postlabeling techniques. The tRNA was digested completely with RNases T1 and A in the presence of alkaline phosphatase, the oligonucleotides were 3'-terminally (3H)-labeled, mapped on PEI-cellulose thin layers, isolated, and sequenced by methods based on base-specific cleavages. Overlaps were obtained by readout sequencing techniques on polyacrylamide gels and PEI-cellulose thin layers. The thin-layer readout technique was used also to locate and identify modified nucleotides. The primary structure was found to exhibit a large degree of homology (94.6%) with silkworm tRNAGCCGly but only 67.6% homology with human tRNACCCGly.


[Is long term medical follow up of tuberculous patients possible? (author's transl)]. Tuberculosis still remains a current disease, of difficult eradication as shown by numerous studies and epidemiological surveys. The authors studied 180 newly affected tuberculous patients admitted to Pr. Laval's department at Marseille and tried to discover why only 91 (50.6%) were still followed or could be traced 3 years after their first hospitalization. Five parameters were studied: origin, sex and age, living area, socio-economical data (change of address, lack of accomodation, profession), tobacco and alcohol addiction. No significant part was played by nationality, living area, or profession. On the other hand, changing of address, chronic tobacco and alcohol addiction were the factors responsible for loosing sight of the patients. An attempt to define this tuberculous patient, which would not be followed up for long, gave the picture of a 40 to 45 years old man, living in a hotel or with no fixed accomodation, smoking more than 20 cigarettes a day and drinking. Such a profile of a tuberculous patient should increase the watchfulness over likely ones.


The effect of organic nitrites and nitrates on glycogenolytic activity in the heart and skeletal muscle. Organic nitrites (sodium nitrite, amyl nitrite) and nitrates (nitroglycerin, pentaerythritol, nitromannite) affect similarly phosphorolytic, amylolytic and total glycogenolytic activities in the heart and skeletal muscle. The action of these drugs on glycogen metabolism is due to liberation of adrenaline and concurrent inhibition of its effect on enzymatic activities. The drugs antagonize the stimulatory action of adrenaline on phospholytic activity and inhibitory action on amylolytic activity.


Glucuronidation of lipophilic substrates: preparation of 3-benzo[a]pyrenyl-beta-D-glucopyranosiduronic acid in multimilligram quantities by microsomal UDP-glucuronyl transferase. A convenient method for the enzymic conversion of multimilligram quantities of 3-hydroxybenzo[a]pyrene to 3-benzo[a]pyrenyl-beta-D-glucopyranosiduronic acid in 90% yield is described. Commercially available freeze-dried rabbit liver microsomes were incubated in the presence of UDPGA, 3-hydroxybenzo[a]pyrene, and Triton X-100 detergent (Figure 1). The course of the biosynthetic reaction was followed by fluorimetry. The glucuronide product was extracted from the acidified incubation supernate with ethyl acetate and the acid function of the glucuronide was utilized in an acid-base extraction procedure to purify the glucuronide from biological and unreacted starting material. The glucuronide precipitated from ethyl acetate and was collected by centrifugation. High pressure liquid chromatography and spectroscopic techniques were used to verify the structure and purity of 3-benzo[a]pyrenyl-beta-D-glucopyranosiduronic acid.


Large scale isolation of pig muscle phosphoglucose isomerase. A large-scale purification procedure for phosphoglucose isomerase from pig skeletal muscle is described. It consists of two fractionations by selective precipitation and two ion exchange chromatography steps yielding an end product of approximately 900 units (micromoles of substrate converted to product per min per mg of protein, at 30 degrees) specific activity. The method separates three isoenzymic forms with an overall recovery of about 30% of the original total enzyme activity in the form of Isoenzyme III, the latter being the predominant enzyme species.


Subtilisin and cyanogen bromide cleavage products of fibronectin that retain gelatin-binding activity. The gelatin-binding region of fibronectin has been obtained by subtilisin digestion and cyanogen bromide cleavage of the molecule. Enzymatic digestion yielded two fragments of molecular weights 50,000 (S50K) and 30,000 (S30K) which were isolated by elution from gelatin-Sepharose affinity columns. Because the S50K fragment also mediated the adhesion of fibroblasts to collagen, it contains both the collagen and cell binding sites on the fibronectin molecule. Both fragments had valine as the NH2-terminal residue, were enriched in half-cystine and methionine residues compared to the whole molecule, and were identical by immunodiffusion. The S50K fragment begins with the sequence Val-Tyr-Gln-Pro-Gln-Pro-His-Pro-Gln-Pro-(Pro)-(Gly)-Tyr-Gly-His-( )-Val, a region with an extended conformation which is susceptible to proteolysis and connects this domain to the remainder of the fibronectin molecule. The S50K fragment appears to be located in the COOH-terminal one-third of the fibronectin molecule but does not contain the interchain disulfide bridge(s); the S30K fragment is probably derived from the NH2-terminal region of S50K.


Control features within the rplJL-rpoBC transcription unit of Escherichia coli. Gene fusions constructed in vitro have been used to examine transcription regulatory signals from the operon which encodes ribosomal proteins L10 and L7/12 and the RNA polymerase beta and beta' subunits (the rplJL-rpoBC operon). Portions of this operon, which were obtained by in vitro deletions, have been placed between the ara promoter and the lacZ gene in the gene-fusion plasmid pMC81 developed by M. Casadaban and S. Cohen. The effect of the inserted DNA segment on the expression of the lacZ gene (in the presence and absence of arabinose) permits the localization of regulatory signals to discrete regions of the rplJL-rpoBC operon. An element that reduces the level of distal gene expression to one-sixth is located on a fragment which spans the rplL-rpoB intercistronic region. This strongly supports the idea that there is an attenuator in this region. The terminator for the operon is located on a fragment which spans the 3' end of the rpoC gene. The major promoter for the operon precedes the rplJ gene [Yamamoto, M. & Nomura, M. (1978) Proc. Natl. Acad. Sci. USA 75, 3891-3895 and Linn, T. & Scaife, J. (1978) Nature (London) 276, 33-37] and was not examined in this study. However, a weak promoter is observed on the fragment that spans the rplJ-rplL intercistronic region. Other regions of the operon may also contain weak promoters. The contribution of these elements to the regulation of this complex operon is discussed.