[Effects of anterior hypothalamic deafferentation on plasma LH response to LH-RH in the baboon (author's transl)]. We reported previously that a biphasic LH release (within 30 min and 90 approximately 150 min after LH-RH injection) was observed in normal menstruating baboons (non-human primates). Plasma immunoassayable LH-RH reached a maximum within 4 min after injection and was undetectable within 60 min. Plasma estrogen and progestin were elevated within 45 min. No plasma LH release with increment of plasma estrogen and progestin was observed in saline injected baboons. This study was performed to investigate the mechanism of biphasic LH release by LH-RH injection. Synthetic LH-RH (100 microgram) was injected sc into four female retrochiasmatic deafferented baboons. Blood samples collected 30 and 2 min before injection and 5, 10, 15, 30, 45, 60, 90, 120, 150 and 180 min after injection were assayed for LH, estrogen and progestin. In four baboons plasma LH peak was observed within 30 min after injection and plasma estrogen and progestin were elevated within 45 min. However, plasma LH peak within 90 approximately 150 min was not observed. These results infer that exogenous LH-RH exerts an effect on the pituitary to release LH within 30 min after injection. Increased estrogen and progestin exert the effect on the higher brain area (extrahypothalamic area) and the anterior hypothalamus to facilitate release of endogenous LH-RH, which subsequently release LH within 90 approximately 150 min in cooperation with exogenous LH-RH which may be considered to participate LH synthesis in the pituitary. Thus, it seems likely that the extrahypothalamic area (for example; limbic system) and the anterior hypothalamus have an important role in regulating LH-RH secretion and subsequent LH release in the baboon.


Total skin electron irradiation in mycosis fungoides. Total skin irradiation with fast electrons (4 MeV, 3,500 rad) was applied in 10 cases of histologically proven mycosis fungoides without any signs of internal dissemination. The technique is described. Complete disappearance of both lesions and symptoms occurred in all patients after treatment. However, half of the patients relapsed after the initial irradiation and were subsequently treated with a booster dose of 1,000 rad 4 MeV electrons or with topical nitrogen mustard. Irradiation with fast electrons seems to be of great importance in the treatment of mycosis fungoides.


The biotransformation of phenylbutazone (Butazolidin). In the present paper a review of published work on the metabolic fate of phenylbutazone is given, inclusive of results from very recent radiotracer studies in man. Phenylbutazone, 1,2-diphenyl-3,5-dioxo-4-n-butylpyrazolidine, being a highly lipophilic compound, is readily absorbed following oral administration to man, and is re-eliminated mostly in metabolized from via urine and faeces. In the biotransformation of phenylbutazone in the human organism two major pathways are involved, i.e. straightforward C-glucuronidation at the 4-position of the pyrazolidine ring and, to a much lesser extent, hydroxylation at one of the phenyl rings or at the n-butyl side chain. Hydrolytic cleavage of the amide bonds in the dioxopyrazolidine ring which, theoretically, would be a possible third pathway of biotransformation, does not take place to any significant extent. In contrast to man, the animal species studies, namely rat and dog, metabolize phenylbutazone predominantly through hydroxylation. This principal difference in metabolic handling may explain why the elimination rate of the drug from plasma is largely different in these species, the average half-lives being 3-4 hours in rat and dog as compared to about 80 hours in man following single doses. The enzyme systems that are primarily involved in phenylbutazone metabolism are mono-oxygenases in the animal species considered, and glucuronyltransferase in man. Auto-induction of enzymes, resultiing in an enhanced elimination of the drug from plasma in the course of repeated administration, has unequivocally been proven for rat and dog only but not for man.


Pharmacology of danazol.

Pharmacology of danazol. We have established that danazol is an orally active pituitary gonadotrophin inhibitory agent devoid of oestrogenic and progestational activity. This steroid has weak, impeded androgenic activity. Danazol is unique in possessing such a profound separation of pituitary gonadotrophin inhibitory activity from overt sex hormone activity. Separate studies have shown that danazol is antiandrogenic, anti-progestational and antioestrogenic. Extensive toxicological studies have established the safety of this steroid. Lastly, the metabolic studies provide convincing evidence that danazol is the active steroid.


Effects of maternal mobility, partner, and endocrine state on social responsiveness of adolescent rhesus monkeys. The social behavior of rhesus monkeys raised for the 1st year of life with mobile (MS) or stationary (SS) cloth surrogate mothers was investigated when the animals reached 4-5 yr of age. The MS males generally refrained from social interaction during initial pairings with females, whereas SS males interacted frequently, but were more often the targets of attacks and chases from adult females than were MS males. The MS males were more likely to vary their social behavior according to the behavior of the social partner and seemed to benefit more from extended social exposure than their SS counterparts. The MS females were more similar to wild-born females than were SS females in nearly every behavior category and dimension tested. These results suggest that rearing with mobile artificial mothers improves the chances of later adaptive social adjustments in socially restricted monkeys.


Olfactory discrimination of squirrel monkey mothers by their infants. Infant squirrel monkeys, 8 to 24 weeks of age, were tested for their ability to discriminate their own mother from another lactating mother by means of olfactory cues, visual cues, or a combination of both. Discrimination by olfactory cues was unequivocal. Addition of static visual cues did not enhance discrimination, and infants showed little evidence of discriminating the mother by means of static visual cues alone. These results suggest that olfaction may be a more significant factor in primate early attachment than has generally been assumed.


Potentiation of glibenclamide-induced insulin release by calcium infusion. The effect of calcium on glibenclamide-induced insulin release was studied in 14 diabetic patients. Two mg glibenclamide was given intravenously and calcium, blood glucose and IRI were determined in venous blood samples at predetermined intervals. The test was repeated 3-4 days later with the patients simultaneously receiving a calcium infusion into a contralateral vein. The decrease in blood glucose and the rise in IRI level were both significantly greater in the combined glibenclamide-calcium test. It is concluded that calcium may temporarily improve carbohydrate tolerance in diabetic patients by potentiating the glibenclamide-stimulated insulin secretion.


Non-clostridial gas gangrene in the diabetic lower limb. Three cases of non-clostridial gas gangrene in diabetic lower limbs are reported. This rare condition occurs only in diabetes with peripheral neuropathy and is invariably fatal unless treated surgically. The clinical picture is deceptive with a slow, but insidiously progressive course, and the late appearance of skin necrosis. Antibacterial therapy is ineffective in preventing death. A serious delay may be fostered by inconclusive bacteriological investigations. Urgent radical amputation is required to save the patient's life.


Migrating myoelectrical complex of the small intestine. An intrinsic activity mediated by the vagus. In healthy conscious parenterally fed dogs and in sheep on their usual diet, the basic motor profile on the small intestine consists of recurring cycles of action potential activity. This cyclic pattern called the migrating myoelectrical complex comprises two distinct phases termed irregular, and regular spiking activity and is followed by a period of quiescence. The pattern persists after bilateral transthoracic vagotomy and in animals rendered diabetic by alloxan. In dogs, feeding disrupts the migrating myoelectrical complex pattern by obliterating the phases of regular spiking activity and quiescence for 6 to 12 hr, depending upon the amount of dry matter intake. After vagotomy a latency in the disruption of the migrating myoelectrical complex pattern with feeding occurs and in vagotomized dogs rendered diabetic, the duration of disruption is strongly reduced. In sheep, the duration of irregular spiking activity of a jejunal segment increased or decreased corresponding to the bulk of digesta, a phenomenon damped after vagotomy. The results indicate that the role of the vagus is limited to (1) prompting prandial disruption, which is then maintained by hormonal effect; (2) regulating the irregular spiking activity duration in relation to the bulk of digesta.


Role of biliary lecithin in lymphatic transport of fat. This study was undertaken to asses the role of luminal lecithin in the lymphatic transportation of fat as chylomicrons. Two doses of fat, the low and high dose, were fed to two different groups of rats, control and bile fistula. At low dose, infusing at 35 mumoles of total fatty acid per hr of a mixture of oleic acid and monoolein, molar ratio 2:1, solubilized in 55 mumoles of sodium taurocholate, there was no difference in the lymphatic output of absorbed fat during steady state (7th and 8th hour) absorption. Infusing at a high dose, 173 mumoles of total fatty acid per hr of a mixture of oleic acid and monoolein, molar ratio 2:1, solubilized in 55 mumoles of sodium taurocholate, the bile fistula rats had lower triglyceride and phospholipid output, with a higher proportion of oleic acid in lymph lecithin than did control rats. These alterations in bile fistula rats returned to normal by addding 10 mumoles per hr of biliary lecithin to the infusate. We conclude that intraluminal biliary lecithin plays a significant role in the translocation of high doses of absorbed fat into lymph and in the amount and type of lecithin synthesized.


Location and underreplication of satellite DNA in Drosophila melanogaster. The two light nuclear satellites (PCsC1 = 1.672 and PCsC1 = 1.687) have been quantified in DNA isolated from the larvel imaginal discs and brains of Drosophila melanogaster with the genotypes X/O, X/X and X/Y. By comparing the results from these different genotypes, the amounts of the two satellites in the X and Y chromosomes and in the autosomes have been determined. The lightest satellite is not located to any appreciable extent in the X chromosome. The heterochromatic regions are not completely filled by these satellites. --Satellite DNA has also been quantified in DNA isolated from adults containing different genotypes. The two satellites are underreplicated to different extents. The apparent amount of underreplication for one of the satellites is different in different parts of the genome.


Induced dominant lethal mutations and cytotoxic effects in germ cells of Drosophila melanogaster with Trenimon, PDMT and sodium monofluorophosphate. Male and female Drosophila melanogaster with special sex chromosome or special autosome constitutions were fed with the mutagenic chemicals Trenimon (2,3,5-trisethyleneimino-1,4-benzoquinone) and PDMT (1-phenyl-3, 3-dimethyltriazene) and with the toxic substance Na2PO3F (sodium monofluorophosphate). The frequency of dominant lethality was recorded among the progeny. The results clearly show that dominant lethality is dose dependent for Trenimon- or PDMT-treated chromosomes in mature sperm and mature oocytes, and an increased amount of chromosomal material per nucleus yields an enhanced lethality. In contrast, a pure toxic effect of Na2PO3F on mature oocytes was demonstrated with one type of female. --With the stocks of Drosophila used, it is possible to distinguish between mutagenic and toxic effects of chemicals on the germ cells. Therefore, dominant lethality can be used as a simple and quick screening test for chemical mutagens.


[W-reactivation and W-mutagenesis in UV-irradiated phage phil05 of Bacillus subtilis]. The survival of UV-irradiated phage ø105 on the lawns of several strains of Bacillus subtilis: wild type (strain 168) and 11 recombination-defficient mutants (recA1, recB2, recB3, recB19, recD27, recF15, recF18, recK4, recM13, recL16 and recO61) was investigated. All rec mutants have the phenotype Hcr+, i.e. normally host-cell reactivate UV-damaged phage. Small doses of UV-irradiation given to the wild type (rec+) cells increase the probability of survival of UV-irradiated ø105 phage (W-reactivation) and significantly enhance the frequency of c-mutants (W-mutagenesis). Maximal frequency of clear mutations in conditions of W-mutagenesis is 3-10(-3), i.e. is 100 times higher than the spontaneous background. Various rec mutations of host cells only diminish the level of W-reactivation but do not eliminate it completely. The most deficient in W-reactivation is recD27 mutant. Mutations recB2, B3, B19 and O61 have no effect on W-mutagenesis of UV-irradiated phage ø105 and on UV-induction of ø105, F15,F18 and L16 mutants. UV-irradiation of lysogenic cells of these mutants does not induce ø105 prophage.


[Escherichia coli K-12 mutants capable of catabolizing purine nucleosides in the absence of purine nucleoside phosphorylase]. Strains of Escherichia coli K-12 defective in purine nucleoside phosphorylase (pup gene) formed on the medium with inosine as the source of carbon and energy phenotypical reversions for the ability of utilizing inosine as source of carbon or purines. The phenotypical suppression of the purine nucleoside phosphorylase deficiency is the result of the mutations (called pnd), which are mapped on the chromosome of E. coli beyond the region of the structural pup-gene location and have phenotypic manifestation distinct from that of pup+ allele: a) pnd mutants divide into some groups for the ability of utilizing several purine nucleosides, including xantosine that cannot be metabolized by pnd+ strains of E. coli; b) pnd mutations do not restore the ability of purine auxotrophs (pur) defective in purine nucleoside phosphorylase (pup) and adenine phosphoribosyltransferase (apt) to grow on the medium with adenine as the sole source of purines. Cell-free extracts of pnd mutants fail to degrade the guanine nucleosides in the absence of phosphate or arsenate ions. These data (and also the ability of pnd mutants to utilize both purine ribonucleosides and deoxyribonucleosides) seem to indicate that the activities induced by pnd mutations are phosphorylase activities.


[UV-mutagenesis in Bacillus subtilis. VII. Induction of auxotrophic mutants]. An experimental testing of material from thin-layered, transparent in passing light, colonies which appear with some frequency after plating Bacillus subtilis cells on agar medium with limited enrichment, has shown that such colonies are formed by auxotrophic mutants. The growth requirements for many of them has been identified. The most of mutants can be reversed to original phenotype by UV-irradiation. The frequency of auxotrophs increases after UV-irradiation of suspension of original cells. The sensitivity of auxotrophic mutants to inactivating action of UV-light is near to that of original cells, hence the increase of the frequency of mutants with dose is a result of induction, but not of selection of preexisting spontaneous auxotrophic mutants. The frequency of induced auxotrophs, in contrast to that of suppressor revertants, badly give way to declining in the time of temporary inhibition of postradiation growth. In the case of Bac, subtilis, the system of induced auxotrophic mutants on the medium with limited enrichment is rather comfortable in use and can be recommended for studying UV-induced mutagenesis in structural genes as well as for testing mutagenic activities.


Flexor tendon repair in "No Man's Land" an experimental study. Twenty-five fingers of seven monkeys were used for an experimental study of a new concept of flexor tendon repair in "No Man's Land". Microsurgical aids were used to shift distally a sleeve of healthy flexor tendon sheath to cover the site of the repaired flexor tendon. There was a low success rate due to technical difficulties in the immobilisation of the monkey fingers. Where the finger was successfully immobilised the results were encouraging.


[A pharmacological study of chlorphenesin carbamate. Tolerance to chlorphenesin carbamate (author's transl)]. Tolerance to chlorphenesin carbamate (CPC) was investigated from the viewpoints of action of CPC, serum free CPC concentration, the activity of UDP-glucuronyltransferase and the content of cytochrome P-450. CPC was administered once daily for 7 or 14 days. In mice, the hypnotic action of hexobarbital injected 24 hours after the last administration of CPC and the motor incoordinating action of CPC significantly decreased on the 7th day, but slightly recovered on the 14th day. Serum free CPC concentration also decreased on the 7th day, but recovered on the 14th day. A significant relationship between the motor incoordinating action of CPC and serum free CPC concentration was observed. Therefore, the recovery of CPC effect on the 14th day was considered to be due to the recovery from the induction of drug-metabolizing enzymes. On the other hand, in rats, the weekly alteration of the motor incoordinating action of CPC was similar to that observed in mice. Serum free CPC concentration on the 7th and 14th days was lower than that on the 1st day, and enzyme induction was observed during CPC administration. Notwithstanding the low level of serum free CPC concentration, the recovery of CPC effect was observed on the 14th day and such was considered to be due to habituation to the rotarod. In mice and rats, it was demonstrated that the intensity of CPC effect was dependent on serum free CPC concentration to the extent that enzyme induction played an important role in the development of tolerance. From these results, the tolerance to CPC is attributed to induction of drug-metabolizing enzymes in liver microsomes.


Cranial and dental anomalies in three species of platyrrhine monkeys from Nicaragua. This paper presents information on the cranial and dental anomalies observed in the crania of three species of platyrrhine monkeys collected in Nicaragua. Cranial anomalies that are discussed include plagiocephaly, bregmatic fontanelle bones, Wormian bones, cranial trauma, and heterotopic bones. Among the dental anomalies that were studied were the following: crazing caries, periodontal and pulpal disorders, alveolar thinning, mechanical loss, excessive attrition, shear bite, impacted molars, supernumerary teeth, congenital agencies, congenital crown aberration, and irregular placement.


[Behavioural and electrophysiological experiments on the sense of taste in Saguinus midas tamarin (Callitrichidae) (author's transl)]. The activity in the gustatory nerve from the anterior part of the tongue, the chorda tympani proper nerve, has been recorded during stimulation of the tongue of a New World monkey, Saguinus midas tamarin. A series of 0.3 M sugars and 0.07 M NaCl, 0.07 M sucrose, 0.004 M acetic acid and 0.00005 M quinine hydrochloride were used as taste stimuli. The concentrations of the last four stimuli were the same as those earlier found to be the lowest concentrations at which this monkey in behavioural experiments discriminates between them and water. The records showed that these threshold concentrations all elicited a neural response. Further, the neural activities during stimulation with the series of 0.3 M sugars were recorded. Using the amplitudes of the responses as a measure, the order between them was found to be fructose greater than glucose greater than lactose greater than arabinose greater than sucrose = galactose greater than raffinose. This order was discussed and related with the order found in behavioural experiments. With both methods, fructose seemed to be the strongest stimulus, but then the order among the sugars differed.


Effects of geographic transfer on the timing of seasonal breeding of rhesus monkeys. This is a study of the effects of geographic transfer on the timing of seasonal breeding of free-living rhesus monkeys on Cayo Santiago and La Parguera islands in Puerto Rico. The results demonstrated a progressive shift in the breeding season of the transferred group. The full shift required a period of 2 years. Change from one environment to another produced shifts in onset of estrous. The explanation for this phenomenon proposes an interaction between the specific reproductive state of the female, which may or may not be sensitive to a particular environmental change, and the environmental variation which is correlated with onset of breeding.


New species of eocene primates and the phylogeny of European adapidae. Restudy of virtually all of the important collections of European Eocene primates of the family Adapidae indicates that 28 valid species in 8 genera are now known, spanning a time period of nearly 20 million years. The biostratigraphic distribution of each species has been studied in the context of established reference levels, and a maximum of four evolutionary lineages are known from any one locality. Ten new species are proposed in the genera Pelycodus (1 sp.), Protoadapis (4 sp.), Periconodon (2 sp.), Anchomomys (1 sp.), and Adapis (2 sp.). Phylogenetic relationships among the species, based on stratophenetic linking, are indicated whenever possible.


A comparison of the reproductive cycle of three species of Lemur. The duration of the luteal phase and the 'proestrus' phase in three species of lemurs (Lemur macaco, Lemur catta, and Lemur variegatus) was determined by serum progesterone measurement. Total immunoreactive estrogens were measured and remained below 0.20 ng/ml except for an apparent peak of short duration during estrus. Circulating testosterone levels in males confirm previously described seasonal gonadal function. The reproductive cycle of the female lemur is discussed and compared to the cycles  of estrous and menstrual animals.


Reproductive cycles of the talapoin monkey (Miopithecus talapoin). Reproductive cycles of female talapoins living in caged breeding groups were followed over 7 years, using visible changes of the perineum. Changes in the perineum during the extended adolescence of this species are described. There was a clearly defined breeding season, although its onset was not so abrupt as in the wild. During the summer months females show very small perineal swellings or none at all. Even during the breeding season all stages of the follicular phase of cycles are highly variable. Completed menstrual cycles are relatively rare since talapoins are highly fertile; they may be somewhat shorter than fertile cycles. Cyclical activity is modifiable by social stimuli, and it is suggested that differences in synchrony of breeding between wild and captive populations might be due both to indequate climate cues and to a lower level of social stimulation in captivity. The concept of the menstrual cycle had limited value in predicting reproductive activity in this highly fertile seasonal breeder.


Social organization of captive monandrous squirrel monkey groups (Saimiri sciureus). 1 adult male and 4 adult female squirrel monkeys were observed together as a group, isolated from all other monkeys. 3 of the 4 females were deafened for a previous experiment. Deafening, however, had no apparent, permanent effect on social behavior. Social dominance hierarchy was evaluated in a variety of situations. The results were compared with those of a similar set of observations on the females prior to the introduction of the male. Before the male was introduced, the dyadic interactions involving food stealing, body grasping, and sexual behaviors were indicative of a female linear rank order. After the male was introduced, the rank order among the females generally remained intact, with the male becoming the highest ranking member in the group. The noteworthy exception to the stability involved the highest ranking female, whose position in the hierarchy was threatened. Heterosexual interactions predominated. Homosexual behavior was also observed, although appreciably reduced in frequency as compared to the all-female group situation. A similar rank order hierarchy was observed in a second group of squirrel monkeys comprised of 1 adult male and 4 adult females. None of these monkeys was deaf.


Brightness preference in nocturnal and diurnal South American monkeys. In a test situation where choice was not tied to reinforcement, both squirrel monkeys (Saimiri sciureus) and owl monkeys (Aotus trivirgatus) consistently chose to view the more luminant of a pair of lights. Over a two log unit range of luminance, the degree of preference in both species for the brighter of two lights was found to depend on their difference in luminance and not on their actual luminances.


[Adrevil improves the peripheral blood flow. Results of a study using the 133-Xe muscle clearance method on lower limbs]. The efficacy of Adrevil has been investigated in 35 patients suffering from arterial blood flow disorders in the lower limb. The patients were given a daily dose of 3 X 2 tablets over a period of 30 days. Changes in muscle perfusion were determined by means of the 133 Xenon muscle clearance. The perfusion volume was found to be increased by 8--12% depending on the severity of the clinical state before treatment. The effect of the drug was more pronounced in cases with severe pathological changes prior to treatment. No adverse reactions were observed.


[Salmonella infections].

[Salmonella infections]. Inspite of a high standard of hygiene, infections due to salmonellae are increasing in the highly developed countries. In the Federal Republic of Germany 36 000 cases were reported in 1976; the number of unreported cases is probably five times higher. Practical and clinical aspects make it necessary to differentiate between typhoidal, systemic infections and enteric salmonellosis, which normally remains localized in the gut. In the treatment of typhoid fever, co-trimoxazole seems to be a potent drug which can replace chloramphenicole. In cases of enteritis due to salmonellae the most important therapeutic aim is to replace the loss of fluid. This can be achieved in most cases by an oral glucose-electrolyte-solution. Antibiotics are of no value in these cases. The treatment of prolonged excretion of salmonellae with the stool remains a problem. Antibiotics usually fail. Some authors have tried drugs which tend to normalize the bacterial flora of the gut; there are no controlled studies about the effectiveness of this kind of treatment.


[Experimental liver changes by means of polychlorinated biphenyls (PCB)]. Liver changes were studied after acut and subchronic exposition to mixtures of polychlorinated biphenyls (PCB) differing in their average chlorine content. Morphologically, the liver changes consisted mainly of cell hypertrophy and proliferation of the smooth endoplasmic reticulum (SER). After subchronic feeding the SER membranes formed membrane arrays. In addition to that the livers showed a hepatic porphyria. It was found that the porphyria was due to an inhibition of uroporphyrin decarboxylation similar to that caused by hexachlorobenzene. The porphyrinogenic effect was increased by feeding PCB containing more highly chlorinated components. Concomitant with the proliferation of the SER a high induction of membrane bound drug metabolizing enzymes could be observed. Duration and amount of induction increased with the chlorine content of the PCB. This could be correlated to the persistence to highly chlorinated components.


Implications of the Donaldson decision: a model for periodic review of committed patients. In 1976 the Connecticut Valley Hospital, the Yale University School of Law, and the department of psychiatry of the Yale School of Medicine, undertook a project in which all involuntary patients at the hospital, a total of 107, were reviewed by three interdisciplinary panels, each composed of a law student, a psychiatric resident, and a hospital social worker. The major purposes of the project were to identify those patients entitled to release under the Donaldson criteria, to develop a model review procedure, and to assess the need for alternative placements. The panels found no patients resembling Kenneth Donaldson, but they did find 53 per cent of the patients inappropriately detained on involuntary status. However, in most cases the recommendation was transfer to a nursing home because of the patient's high degree of disability. The authors emphaxize the importance of periodic external reviews of commitment status.


Institutions without walls for emotionally disturbed children. Chedoke Child and Family Centre, Hamilton, Ontario. The parent-therapist program was developed as an alternative to residential treatment centers for severely emotionally disturbed children. Five healthy nuclear families function in a group as an extended family. Each family receives a salary and is responsible for the care and protection of an emotionally disturbed child. Mental health professionals provide the parents with education and supervision. By the end of 1976 a total of 36 children ranging in age from six to 15 had been treated by 20 different parent-therapist couples for periods ranging from nine to 26 months. A comparison of the program with those of two residential centers showed that there was no difference in treatment outcome but that costs for the parent-therapist program were half those for the residential centers.


[Seroepidemiological investigations on the prevalence of rickettsial antibodies in man in the Federal Republic of Germany (author's transl)]. 1,600 random serum samples from inhabitants of Hesse were investigated for rickettsial antibodies by complement fixation test. Antibodies were found in 19 samples (1,2%) against RMSF-group antigen with titers between 1:10 and 1:640. These results confirm recent reports on natural foci of such rickettsial infections in the Federal Republic of Germany. Antibodies against rickettsia of the Typhus-group or Coxiella burnetii could not be detected by the complement fixation reaction. None of the sera reacted with significant titer in the Weil-Felix-Test.


[Disodium cromoglycat (intal) in the treatment of allergic states of the respiratory system (author's transl)]. Disodium cromoglycate (Intal) has been found several years ago to act as an reliable drug in the treatment of allergic states of the respiratory system. The effective mechanism seems to be its stabilizing function for circulating and sessile mast cells, thus avoiding the release of histamine and other vasoactive amines after an immunological or non-immunological stimulus. Although DSCG does not inhibit the binding of antigen to its antibody it must be given prophylactically before the antigen has access to the mast cells. Most efficiently DSCG is used in the treatment of the exogenous allergic asthma, and secondly in allergic rhinitis (70-90% success). Own observations confirm the effectiveness of the drug especially in patients with pollinosis. In addition it was found that DSCG can be used during hyposensitization procedures and that the drug does not influence the results of skin testing. At present there have been detected neither side-effects nor interactions with other drugs.


[Clinical and metabolic studies in the acute stage of Crohn's disease with total parenteral feeding and elementary diet]. The acute phase of Crohn's disease in a 14 year old female was effectively treated with total parenteral nutrition (TPN) and an elemental diet (ED). Acute abdominal symptoms subsided at once. With TPN there was a daily nitrogen retention of 3,2-5.6 grams. With ED nitrogen retention decreased presumably because of the reduced nitrogen content of the oral diet. During the 7 weeks of treatment body weight increased 7 kgrams. Observed skin lesions were caused by zinc deficiency.


Usefulness of transtracheal puncture and aspiration in the bacteriological diagnosis of pulmonary tuberculosis. Transtracheal puncture enables two samples of bronchial secretions to be taken-the product of transtracheal aspiration and simultaneously expectorated sputum (obtained in 71% of the cases)-for the purpose of testing for Mycobacterium tuberculosis in cases of suspected pulmonary tuberculosis. Two groups of patients were studied: Group I: 100 patients who were poor expectorators and who all underwent transtracheal puncture; Group II (Control): 100 patients who expectorated well or who had been given gastric lavages immediately on admission. Laboratroy analyses revealed M. tuberculosis in at least one of the samples obtained from each of the 200 patients. The authors compare the efficiency of the methods used within each group and between the two groups. Samples obtained by transtracheal aspiration and simultaneous expectoration (75% of positive results) more often contained M. tuberculosis than the other Group I samples (64% of total positive results; 64% of positive results for spontaneous sputa, 65% for gastric fluids), and as often as the Group II samples (76% of total positive results), particularly the expectoration samples (78% of positive results). Simultaneously expectorated sputum more frequently contained M. tuberculosis (82% of positive results) than transtracheal aspiration (69% of positive results). Transtracheal puncture and/or simultaneous expectoration were the only examinations revealing M. tuberculosis in 34 patients in Group I. Non-specific bacteriological findings are not relevant. However, the authors point out that this technique is not always innocuous (although no serious complications were observed in this series), and that transtracheal puncture must always be carried out by physicians trained in the technique.


You are dangerous to your health: the ideology and politics of victim blaming. This article describes the emergence of an ideology which blames the individual for her or his illness and proposes that, instead of relying on costly and inefficient medical services, the individual should take more responsibility for her or his health. At-risk behavior is seen as the problem and changing life-style, through education and/or economic sanctions, as the solution. The emergence of the ideology is explained by the contradictions arising from the threat of high medical costs, popular expectations of medicine along with political pressures for protection or extension of entitlements, and the politicization of environmental and occupational health issues. These contradictions produce a crisis which is at once economic, political and ideological, and which requires responses to destabilizing conditions in each of these spheres. These ideological initiatives, on the one hand, serve to reorder expectations and to justify the retrenchment from rights and entitlements for access to medical services, and, on the other, attempt to divert attention from the social causation of disease in the commercial and industrial sectors.


Clinical studies of the interaction between tolmetin and glibenclamide. Forty patients with diabetes mellitus and rheumatic disorders (rheumatoid arthritis, osteoarthritis, soft-tissue rheumatism) were included in a double-blind trial with the purpose of investigating whether the concomitant administration of tolmetin and glibenclamide leads to an interaction. No significant alterations of blood and urine glucose values could be found in comparison to placebo. Therefore it can be assumed that there is no interaction between these two substances.


Condylar fractures of the mandible. IV. Function of the masticatory system. The masticatory system was followed-up with respect to symptoms of dysfunction such as restricted mandibular movements, tenderness, and clicking from muscles and joints at predetermined times after condylar fractures of the mandible in 24 children, 17 teenagers, and 26 adults. Two years after the trauma, the initially reduced (less than 40 mm) maximal opening became normal (larger than or equal to 40 mm) in all but one individual. Asymmetry of mandibular movements, such as deviation at maximal opening and differences between fractured and nonfractured side in maximal lacterotrusion and protrusion, usually disappeared within 2 years in children, while it persisted or became aggravated in adults. Persistent symtoms from the joints and muscles, such as clicking and tenderness, were rare in children and frequent in adults.


Nutritional deficiency in oral candidosis. A retrospective study of 108 patients was carried out to investigate the possible relationship between infection of the mouth with Candida albicans and blood levels of iron, folic acid and vitamin B12. The patients were separated into two groups--those with hyperplastic and those with atrophic candidal lesions--and compared with separate control groups. Twenty-one patients had chronic hyperplastic candidosis and seven were iron deficient. Comparison with an age- and sex-matched control group showed the differences to be significant only at the p less than 0.1 level. Seven of the patients with hyperplastic lesions had folic acid deficiency and the difference between patients and controls was statistically significant (less than 0.05). However, no significant differences in iron or folic acid deficiency were noted between 87 patients with atrophic candidosis and 65 conttrols, and vitamin B12 deficiency was not statistically significant for either the hyperplastic or the atrophic group. It is concluded that deficiency of iron, folic acid or vitamin B12 alone does not promote growth of Candida albicans on the oral mucous membrane but that in some susceptible individuals, iron or folic deficiency may facilitate epithelial invasion by hyphae of Candida albicans.


Value and limitation of panoramic radiography in the diagnosis of maxillary sinus pathosis. Panoramic radiography is superior to the Water's projection for the detection of maxillary sinus malignancy and cyst-like lesions near the floor of the maxillary sinus. Panoramic radiography is, however, not a technique for the examination of cloudiness, and thickening of the mucous membrane of the maxillary sinus, and bony sclerosis of the maxilla associated with maxillary sinusitis. The two techniques supplement each other in the diagnosis of maxillary sinus pathosis.


Cultivation of periodontal ligament fibroblasts on extracted monkey incisor. A histologic study of three culturing methods. Forty-two permanent incisors were extracted from green vervet monkeys and cultivated in Eagle's medium at 37 degrees C using the following culture methods: 1. teeth placed at the bottom of the culture flask; 2. teeth placed in an upright position in the culture flask and supported by an open glass cylinder; 3. teeth placed in an upright position in the culture flask and supported by a semisolid tissue culture medium. The teeth were cultured from 1 to 21 days and then examined histologically and periodontal changes registered histometrically. After 2 days, most of the periodontal membrane showed signs of extensive cell necrosis. After 3 days the first signs of proliferation of cells were seen either as clusters around existing vessels or as a surface layer on the periodontal membrane. After 7 days an almost complete over of connective tissue cells had formed on the root surface. This layer had formed on periodontal ligament with cell necrosis as well as on areas with preserved vitality of the periodontal cells. Areas on the root surface without periodontal ligaments were also covered with a cell layer. No significant change took place within the periodontal membrane with extended culture periods (2 and 3 weeks). Better results were achieved using liquid medium as compared with semisolid medium.


Liposarcoma of the buccal mucosa.

Liposarcoma of the buccal mucosa. An extremely rare tumor of the oral cavity, liposarcoma, is reported. The tumor was located in the submucosal layers of the cheek and protruded into the mouth. To the best of our knowledge this is the fourth reported instance of liposarcoma of the cheek in the world literature. Combined surgical excision and radiotherapy proved a success as judged by a 3-year follow-up.


Ultrasonography in the diagnosis of a cystic hygroma. The use of ultrasonography in the diagnosis of soft tissue lesions is described and exemplified in a 33-year-old female with a cystic hygroma in the submandibular region.


Cyanoacrylate spray in the treatment of prolonged oral bleeding. n-Butyl-cyanoacrylate is an effective hemostatic agent. Optimal efficiency depends on local application via a spray to achieve a thin and elastic film. Construction of a spray applicable to the oral cavity is illustrated using a plastic ampule with Histoarkryl as reservoir. This spray has been used in 27 cases of prolonged bleeding to achieve hemostasis. In 18 cases the patients suffered from different hemorrhagic diatheses and general hemostatic procedures would normally have been necessary to achieve hemostasis. In nine patients without hemorrhagic diathesis, the cyanoacrylate spraying replaced more complicated local procedures. In 24 cases the cyanoacrylate spraying was a sufficient hemostatic treatment.


Studies with an improved white cell isolation method to assess the vitamin C status in surveys. A method for isolating white cell which employs a disposable syringe has been modified and adapted to nutrition survey work. Methylcellulose was used as a clumping agent for erythrocytes. This method was more rapid and was easier to perform than a previously recommended metrizoic acid technique which gave similar results. Average leucocyte recovery was 62% and erythrocyte contamination was 0.75 erythrocyte per leucocyte. Recovery of added ascorbic acid to white cell pellets was better than 95% and tests of the reproducibility of the assay gave a coefficient of variation of 6% for samples in the deficient range. Extraction of vitamin C from white cells was carried out in a centrifuge tube and took only 30 sec. Vitamin C was found to be stable for several weeks after freezing either the white cell extracts in dilute metaphosphoric acid or simply the white cell pellets. The method was utilized under field conditions during a small survey of Eskimos from Arctic Bay. The significance of the white cell vitamin C content as an index of nutritional status is discussed.


TRH in depressive illness.

TRH in depressive illness. Among the four hypothalamic-releasing hormones (TRH, SRIF, LHRH, and MIF) with central nervous system actions and possibly with psychotropic properties, TRH is the one most systematically studied. Animal pharmacological data has confirmed the activity of TRH as a centrally acting agent with a widespread neurophysiological role independent of the pituitary-thyroid axis; and human pharmacological studies revealed that the computer EEG profile of TRH resembles psychostimulant drugs and secondary amine tricyclic antidepressants. Corresponding with this are the clinical findings that TRH produces a rapid but transient improvement in unipolar depression in women, a sense of increased energy and well-being, or increase of interest, desire and drive for work, food, and sex. In spite of the equivocal findings in some of the clinical studies, a recent investigation raised the possibility that a transient favorable response to TRH may identify a patient population responsive to maprotilline, or other norepinephrine uptake inhibitors.


Uveoscleral aqueous outflow in the rhesus monkey: importance of uveal reabsorption. The uveal absorption of aqueous humor at two different intraocular pressures was measured in rhesus monkeys by sampling vortex vein blood during anterior chamber perfusion of Ringer's solution containing both fluorescein and 125I albumin. At 20 mm. Hg, an excess of fluorescein equivalent in amount to 0.45 microliter/min. of anterior chamber perfusate and an excess of 125I albumin equivalent to 0.18 microliter/min. of perfusate was found in the vortex vein blood compared to systemic blood. At 32 mm. Hg, an excess of 0.86 microliter/min. of fluorescein and 0.26 microliter/min. of 125I albumin was measured. The increase in fluorescein absorption at the higher intraocular pressure was significant (0.025 less than p less than 0.05), with a uveal outflow facility of 0.034 microliter/min./mm. Hg. At an intraocular pressure of 20 mm. Hg, uveal aqueous outflow is less than 10% of total aqueous outflow. The pressure dependence of the uveal uptake of fluorescein implies an aqueous reabsorption into the uveal vessels by ultrafiltration. Uveoscleral aqueous "outflow" appears to consist of intraocular uveal reabsorption of water and small molecules from the aqueous, with only larger molecules like albumin actually leaving the eye through posterior scleral vessel perforations. This reabsorption plays a minor role in intraocular pressure regulation under normal conditions.


Ocular abnormalities in the myopathic hamster (UM-X7.1 strain). Eyes from cardiomyopathic hamsters (UM-X7.1 strain) were examined histologically for evidence of ocular defects. Changes observed included microphthalmia, scleral ectasia, scleral rupture, keratoconus, retinal detachment, retinal dysplasia, retinal fragmentation, retinal thinning, fibrosis of iris and ciliary body, ectopia lentis, and cataract formation. Lesions characteristic of cardiomyopathic hamsters were observed in the myocardium and skeletal muscle. This strain may be a suitable animal model to study the pathogenesis of ocular changes seen in certain congenital connective tissue disorders in man.


Vitreal syneresis in rhesus monkeys.

Vitreal syneresis in rhesus monkeys. The eyes of 15 rhesus monkeys were evaluated. Various degrees of vitreal syneresis were observed in 28 of the 30 eyes. The observed vitreal structures varied from fine strands randomly spaced throughout the vitreous to thick, intertwining, fibrous networks with some clumping of the collagenous condensate at the fiber junctions. Qualitatively, the degree of syneresis was slightly more extensive in the eight older mature males than in the seven younger animals. In all animals a clear view of the fundus could be obtained with the ophthalmoscope. The vitreous structures may be one cause of variability in ocular dose-response relationships for exposure to laser radiation. The effect on retinal exposure experiments of the finer vitreal structure is considered minimal.


An allele-specific suppressor of white-coral in Drosophila melanogaster. A new allele of white-coral (wco2) was isolated from Canton S after mutagenesis. Many common laboratory stocks were found to carry a suppressor gene (Su(wco2)) which alters the phenotype of wco2 flies toward wild-type. The Su(wco2) is allele-specific (it does not suppress wco), dominant, homozygous viable, located near Su(bwV1) on the right arm of chromosome 2, and shows a simple gene-dosage effect. The degree of suppression is sensitive to the genetic background. There appears to be selection for Su(wco2) in a genotype where it does not affect eye pigmentation.


Cytochemical studies of GERL and its role in secretory granule formation in exocrine cells. The structure and cytochemistry of GERL was studied in several different exocrine secretory cells, including the exorbital lacrimal gland, parotid, lingual serous (von Ebner's), submandibular, and sublingual salivary glands, and exocrine pancreas of the rat; the lacrimal, parotid and pancreas of the guinea-pig; and the lacrimal gland of the monkey. GERL was morphologically and cytochemically similar in all cell types studied. It was located in the inner Golgi region and consisted of cisternal and tubular portions. Immature secretory granules were in continuity with GERL through multiple tubular connections. Modified cisternae of endoplasmic reticulum, with ribosomes only on one surface, closely paralleled parts of GERL. GERL and immature granules were intensely reactive for acid phosphatase activity, while the inner Golgi saccules were reactive for thiamine pyrophosphatase and nucleoside diphosphatase activities. In the rat exorbital lacrimal and parotid glands, reaction product for endogenous peroxidase, a secretory enzyme, was present in the endoplasmic reticulum, Golgi saccules, immature and mature secretory granules. GERL was usually free of reaction product or contained only a small amount. The widespread occurrence of GERL in secretory cells, and its intimate involvement with the formation of granules, suggest that it is an integral component of the secretory process.


Ultrastructural cytochemistry of the human adrenal medulla. A cytochemical study of the human adrenal medulla showed that it is made up of two types, the adrenaline (A-) and noradrenaline (N-) storing cells. A-and N-storing granules were argentaphobic when ultra-thin sections of Araldite-embedded medulla were stained according to the periodic acid-thiocarbohydrazide silver proteinate technique of Thiery. A small amount of glycogen (which disappeared after digestion with alpha amylase) in the form of B-particles, as well as lysosomes were, however, visualized by this technique. The entire core of A granules was markedly positive after ultrathin sections of glutaraldehyde-fixed, glycol methacrylate (GMA-) embedded medullae were stained with phosphotungstic acid (PTA) at a low pH (O.3). The N granules, in contrast, were mostly unreactive. PTA stained a large part of the Golgi complex of A cells, whereas it generally had no such effect on that of the N cells. In both cell types, the cell coat, lysosomes and multivesicular bodies reacted to PTA. The periodic acid schiff (PAS) technique showed A but not N granules in semithin sections of GMA-or Araldite-embedded medullae. The PTA and PAS stains were abolished by acetylation, restored by saponification, unchanged by methylation and greatly diminished by sulfation or by digestion with beta glucuronidase after oxidation by perchloric acid. These results indicate that in man the A granules and the Golgi complex of A cells, unlike the same structures in N cells, are rich in glycoproteins.


Extended survival of pancreatic islet allografts in the testis of guinea-pigs. 1. Pancreatic islets, isolated from the guinea-pig pancreas by microdissection and transplanted as allografts to the testis, may survive for 11 weeks, some 5 times the maximum survival of orthotopic skin grafts. 2. Intratesticular islet cell allografts appear not to provoke a systemic immunity such as might be detected by the behaviour of a subsequent orthotopic skin allograft from the same donor. 3. Intratesticular islet cell allografts succumb (in 'second set' fashion) to a pre-existing immunity generated by a prior orthotopic skin allograft. 4. Further experimental work is necessary to determine the relative importance of graft dosage, graft antigenicity, graft site and 'graft adaptation' in determining this extended survival.


Further studies on the transplantation of isolated pancreatic islets. It is concluded that the extended survival of allografts of isolated pancreatic islets in the testis and in the omentum is probably due to several factors. 1. An element of 'privilege' in both graft sites, of unknown nature. 2. The small amount of tissue transplanted. 3. Weaker antigenicity of ancreatic islet tissue as compared with that of skin.


A two-year experience with cromolyn sodium in children with asthma. A two-year experience with cromolyn sodium in children with asthma drawn from inner city population, is presented. Though the initial response to the drug at six weeks was excellent, the response over a two-year period was only very moderate. The possible reasons for this are discussed.


Functional modifications of the translational system in Bacillus subtilis during sporulation. Extracts of sporulating cells were found to be defective in vitro translation of phage SP01 ribonucleic acid (RNA) and vegetative Bacillus subtilis RNA. The activity of washed ribosomes from sporulating cells was very similar to that of washed ribosomes from vegetative cells in translating polyuridylic acid, SP01 RNA, and vegetative RNA. The S-150 fraction from either vegetative or sporulating cells grown in Difco sporulation medium contained an apparent inhibitor of protein synthesis. The crude initiation factor fraction from ribosomes of sporulating cells was defective in promoting the initiation factor-dependent translation of SP01 RNA. The crude initiation factor preparations from sporulating cells were as active as the corresponding preparations from vegetative cells in promoting the initiation factor-dependent translation of either phage Qbeta or phage T4 RNA by washed Escherichia coli ribosomes. The crude initiation factors from sporulating cells were perhaps more active than those from vegetative cells in promoting the initiation factor-dependent synthesis of phage T4 lysozyme by E. coli ribosomes. The crude initiation factor preparations from either vegetative or stationary-phase cells of an asporogenous mutant showed similar ability to promote the in vitro translation of SP01 RNA.


Mercury and organomercurial resistances determined by plasmids in Pseudomonas. Mercury and organomercurial resistance determined by genes on ten Pseudomonas aeruginosa plasmids and one Pseudomonas putida plasmid have been studied with regard to the range of substrates and the range of inducers. The plasmidless strains were sensitive to growth inhibition by Hg(2+) and did not volatilize Hg(0) from Hg(2+). A strain with plasmid RP1 (which does not confer resistance to Hg(2+)) similarly did not volatilize mercury. All 10 plasmids determine mercury resistance by way of an inducible enzyme system. Hg(2+) was reduced to Hg(0), which is insoluble in water and rapidly volatilizes from the growth medium. Plasmids pMG1, pMG2, R26, R933, R93-1, and pVS1 in P. aeruginosa and MER in P. putida conferred resistance to and the ability to volatilize mercury from Hg(2+), but strains with these plasmids were sensitive to and could not volatilize mercury from the organomercurials methylmercury, ethylmercury, phenylmercury, and thimerosal. These plasmids, in addition, conferred resistance to the organomercurials merbromin, p-hydroxymercuribenzoate, and fluorescein mercuric acetate. The other plasmids, FP2, R38, R3108, and pVS2, determined resistance to and decomposition of a range of organomercurials, including methylmercury, ethylmercury, phenylmercury, and thimerosal. These plasmids also conferred resistance to the organomercurials merbromin, p-hydroxymercuribenzoate, and fluorescein mercuric acetate by a mechanism not involving degradation. In all cases, organomercurial decomposition and mercury volatilization were induced by exposure to Hg(2+) or organomercurials. The plasmids differed in the relative efficacy of inducers. Hg(2+) resistance with strains that are organomercurial sensitive appeared to be induced preferentially by Hg(2+) and only poorly by organomercurials to which the cells are sensitive. However, the organomercurials p-hydroxymercuribenzoate, merbromin, and fluorescein mercuric acetate were strong gratuitous inducers but not substrates for the Hg(2+) volatilization system. With strains resistant to phenylmercury and thimerosal, these organomercurials were both inducers and substrates.


A third beta-galactosidase in a strain of Klebsiella that possesses two lac genes. Klebsiella strain RE1544 contains two lac operons, one on the chromosome and one on a lac plasmid. A mutant of RE1544, in which the lacZ genes of both operons produce no active enzyme, was found to synthesize a beta-galactosidase that hydrolyzes ortho-nitrophenyl-beta-D-galactopyranoside but not lactose. Synthesis of this beta-galactosidase (BGase-III) is induced by lactose but not by isopropyl-1-thio-beta-D-galactopyranoside or methyl-beta-D-thiogalactopyranoside. In both the regulation of synthesis and substrate specificity, BGase-III strongly resembles the ebg0 enzyme of Escherichia coli. Nevertheless, by the criteria of immunological cross-reactivity and subunit molecular weight, BGase-III is not related to the ebg0 enzyme.


Isolation and characterization of fusidic acid-resistant, sporulation-defective mutants of Bacillus subtilis. Fusidic acid-resistant, sporulation-defective mutants were isolated from Bacillus subtilis 168 thy trp. About two-thirds of the fusidic acid-resistant (fusr) mutants were defective in sporulation ability and fell into three classes with respect to sporulation character. The representative mutants FUS426 and FUS429 were characterized in detail. FUS426 [fusr spo (Ts)], a temperature-sensitive sporulation mutant, grew well at 30 and 42 degrees C but did not sporulate at 42 degrees C. FUS429 [fusr spo (Con)], conditional sporulation mutant, grew and sporulated normally in the absence of fusidic acid, but its sporulation and growth rates decreased in the presence of fusidic acid, depending on the concentration of the drug. Although electron microscopic observation showed that both mutants were blocked at stage I of sporulation, the physiological analyses indicate that these mutants belong to the SpoOB class. Both mutants formed a thickened cell wall as compared with that of the parental strain. Genetic and in vitro protein synthesis analyses led to the conclusion that the sporulation-defective character of mutants FUS426 and FUS429 resulted from an alteration in elongation factor G caused by a single lesion in the fus locus. The possible role of elongation factor G in sporulation is discussed.


Metal ion dependence of a heat-modifiable protein from the outer membrane of Escherichia coli upon sodium dodecyl sulfate-gel electrophoresis. One heat-modifiable protein of Escherichia coli outer membrane does not completely change to the high-temperature form in the presence of magnesium ion in sodium dodecyl sulfate solution. When the metal ion complexing reagents ethylenediaminetetraacetic acid, phosphate ion, hydroxyl ion, or the competitive cations Zn2+ or Ca2+ are added to the sodium dodecyl sulfate-solubilized sample of outer membrane, and then the sample is heated to 100 degrees C and recooled to room temperature, the protein is almost completely converted to the high-temperature form. In control samples, or if sodium chloride, magnesium chloride, or manganous chloride are added to these samples and treated the same way, a large amount of the low-temperature form of the protein is preserved. beta-Mercaptoethanol additions gave the same results as the metal ion complexing reagents and may owe its activity in these solutions to metal-binding activity and not to its role as a reducing reagent. We concluded that magnesium ion may be involved with stabilization of the low-temperature form of the protein either by directly binding the magnesium or by mediating interaction with other components of the membrane.


Properties of the beta-lactamase specified by the Pseudomonas plasmid RPL11. The Pseudomonas plasmid RPL11 specifies a beta-lactamase that has properties distinguishing it from other plasmid-mediated beta-lactamases so far described, although it closely resembles plasmid Rms139 beta-lactamase.


Motility of flagellated bacteria in viscous environments. The lowest viscosity that immobilized flagellated bacteria such as Psedomonas aeruginosa, Spirillum serpens, and Escherichia coli was 60 centipoise (cp). Much higher viscosities (1,000 cp and higher) were required to immobilize two flagellated bacteria selectively isolated from nature by methods based on their ability to migrate through agar gels. The latter finding indicates that certain flagellated bacteria have the ability to swim through environments of relatively high viscosity. It is suggested that these flagellated bacteria possess a specialized type of motility apparatus suited to viscous conditions present in their habitats.


Use of argA-lac fusions to generate lambda argA-lac bacteriophages and to determine the direction of argA transcription in Escherichia coli. Fusions of lac genes to the argA operator were constructed, and lambda phages carrying these fusions were isolated and characterized. With the aid of a lambda phage carrying an argA-lac fusion, the direction of argA transcription on the Escherichia coli chromosome was determined to be clockwise.


Regulation of argA operon expression in Escherichia coli K-12: cell-free synthesis of beta-galactosidase under argA control. Regulation of argA operon expression in Escherichia coli K-12 was studied in a cell-free, deoxyribonucleic acid-dependent, enzyme-synthesizing system. lambdaAZ-7 deoxyribonucleic acid, which carries a fusion of the lacZ structural gene to the argA operon so that beta-galactosidase synthesis is under argA regulation, was used as the template. To eliminate extraneous readthrough from lambda promoters, lambda repressor was introduced into the synthesis mixtures by preparing the S-30 component from a strain (514X5a-12-29) that carries a multicopy hybrid plasmid (pKB252) containing the lambdacI gene. Under these conditions beta-galactosidase synthesis was repressed 90% by the arginine repressor when a sufficient concentration of L-arginine was present. This repression could be overcome by escape synthesis when the lambdaAZ-7 deoxyribonucleic acid concentration in the synthesis mixtures was increased. Guanosine 3'-diphosphate-5'-diphosphate stimulated beta-galactosidase synthesis from this template.


Lipiarmycin-resistant ribonucleic acid polymerase mutants of Bacillus subtilis. Lipiarmycin inhibited the activity of deoxyribonucleic acid-dependent ribonucleic acid (RNA) polymerase in vitro. We showed that inhibition was due to interference by lipiarmycin with the activity of sigma-containing molecules of RNA polymerase. Transcription by core enzyme was relatively resistant to the drug, but addition of sigma led to highly drug-sensitive RNA synthesis. We isolated lipiarmycin-resistant mutants of Bacillus subtilis and characterized them genetically and biochemically. Drug-resistant mutants contained an altered RNA polymerase that was resistant to the drug in vitro. By separation and mixed reconstitution of core and sigma fractions of mutant and wild-type RNA polymerase, we showed that lipiarmycin resistance in one mutant strain was a property of the core fraction. Genetic mapping experiments indicated that at least two lpm mutants are located between loci determining rifampin resistance and streptolydigin resistance.


Pseudomonas aeruginosa mutants resistant to urea inhibition of growth on acetanilide. Pseudomonas aeruginosa AI 3 was able to grow in medium containing acetanilide (N-phenylacetamide) as a carbon source when NH4+ was the nitrogen source but not when urea was the nitrogen source. AIU mutants isolated from strain AI 3 grew on either medium. Urease levels in bacteria grown in the presence of urea were 10-fold lower when NH4+ or acetanilide was also in the medium, but there were no apparent differences in urease or its synthesis between strain AI 3 and mutant AIU 1N. The first metabolic step in the acetanilide utlization is catalyzed by an amidase. Amidases in several AIU strains showed altered physiochemical properties. Urea inhibited amidase in a time-dependent reaction, but the rates of the inhibitory reaction with amidases from the AIU mutants were slower than with AI 3 amidase. The purified amidase from AIU 1N showed a marked difference in its pH/activity profile from that obtained with purified AI 3 amidase. These observations indicate that the ability of strain AIU 1N and the other mutants to grow on acetanilide/urea medium is associated with a mutation in the amidase structural gene; this was confirmed for strain AIU 1N by transduction.


Properties of Escherichia coli mutants deficient in enzymes of glycolysis. Physiological properties of mutants of Escherichia coli defective in glyceraldehyde 3-phosphate dehydrogenase, glycerate 3-phosphate kinase, or enolase are described. Introduction of a lesion in any one of the reversible steps catalyzed by these enzymes impaired both the glycolytic and gluconeogenic capabilities of the cell and generated an obligatory requirement for a source of carbon above the block (gluconeogenic) and one below (oxidative). A mixture of glycerol and succinate supported the growth of these mutants. Mutants lacking glyceraldehyde 3-phosphate dehydrogenase and glycerate 3-phosphate kinase could grow also on glycerol and glyceric acid, and enolase mutants could grow on glycerate and succinate, whereas double mutants lacking the kinase and enolase required l-serine in addition to glycerol and succinate. Titration of cell yield with limiting amounts of glycerol with Casamino Acids in excess, or vice versa, showed the gluconeogenic requirement of a growing culture of E. coli to be one-twentieth of its total catabolic and anabolic needs. Sugars and their derivatives inhibited growth of these mutants on otherwise permissive media. The mutants accumulated glycolytic intermediates above the blocked enzyme on addition of glucose or glycerol to resting cultures. Glucose inhibited growth and induced lysis. These effects could be substantially overcome by increasing the osmotic strength of the growth medium and, in addition, including 5 mM cyclic adenosine 3',5'-monophosphate therein. This substance countered to a large extent the severe repression of beta-galactosidase synthesis that glucose caused in these mutants.


Regulation of a common amidotransferase subunit. In Bacillus subtilis the trpX locus specifies a glutamine-binding protein designated subunit X, which forms a complex with subunit E to constitute the anthranilate synthase enzyme aggregate (EX) and subunit A to constitute the p-aminobenzoate synthase enzyme aggregate (AX). Subunit X confers upon these enzyme complexes the ability to utilize glutamine as a substrate. The trpX locus has been examined to determine its map position and control. (i) The trpX locus was found to be cotransformed with the lysS and pabA loci. The results of three-factor transformation analyses suggest the following order of these markers: lysS-sul-trpX-pabA. (ii) Mutation to constitutivity of the tryptophan operon resulted in a 50- to 60-fold increase in the level of subunit X when the mutant contained functional trE and abA gene products; however, in the absence of subunit E there was only a 4- to 5-fold increase in the glutamine-binding protein. (iii) Formation of subunit X was derepressed under conditions that allow for the derepression of the trpE and/or pabA loci. (iv) Subunit X synthesis was derepressed to a greater extent in mutants that contain a functional trpE gene product than in mutants that contain a nonsense mutation in the trpE locus. These results are consistent with the hypothesis that the trpE and pabA gene products affect the expression and control of the trpX locus.


Levels of acetyl coenzyme A, reduced and oxidized coenzyme A, and coenzyme A in disulfide linkage to protein in dormant and germinated spores and growing and sporulating cells of Bacillus megaterium. Dormant spores of Bacillus megaterium were found to contain approximately 850 pmol of coenzyme A (CoA) per milligram of dry weight. Of this total, less than 1.5% was acetyl-CoA, 25% was CoA-disulfide, 43% was in disulfide linkage to protein, and the remainder was the free thiol. Dormand spores of Bacillus cereus and Clostridium bifermentans contained 700 and 600 pmol of CoA per milligram of dry weight, respectively; in both species approximately 45% of the CoA 45% of the CoA was in disulfide linkage to protein. During germination of spores of all three species, greater than 75% of the CoA-protein disulfides were cleaved. In B. megaterium, cleavage of these disulfides during spore germination did not require exogenous metabolites and occurred at about the same time as the initiation of germination. Much of the CoA was converted to acetyl-CoA at this time. Dormant spores also contained reduced nicotinamide adenine dinucleotide-dependent CoA-disulfide reductase at levels higher than those in other stages of growth. The level of total CoA in the growing cells was two- to three-fold higher than in spores. This level remained constant throughout growth and sporulation, but less than 2% of the total cellular CoA was in disulfide linkage to protein until late in sporulation. The CoA-protein disulfides accumulated exclusively within the developing spore at about the time when dipicolinic acid was accumulated.


Bacillus subtilis mutant with temperature-sensitive net synthesis of phosphatidylethanolamine. Bacillus subtilis mutants with temperature-sensitive growth on complex media were screened for defects in phospholipid metabolism. One mutant was isolated that showed temperature-sensitive net synthesis of phosphatidylethanolamine. The mutant did not accumulate phosphatidylserine at the nonpermissive temperature. In the presence of hydroxylamine, wild-type B. subtilis accumulated phosphatidylserine at both 32 and 45 degrees C, whereas the mutant did only at 32 degrees C. In vitro phosphatidylethanolamine synthesis with bacterial membranes is no more temperature sensitive with mutant membranes than with wild-type membranes. The mutation probably affects the synthesis indirectly, possibly by altering a membrane protein. The mutant bacteria grew at the nonpermissive temperature, 45 degrees C, in a phosphate buffer-based minimal medium, although net synthesis of phosphatidylethanolamine was also temperature sensitive in this medium. One mutation caused both temperature-sensitive growth on complex media and temperature-sensitive net synthesis of phosphatidylethanolamine. The mutation is linked to aroD by transformation.


Cleavage of adenosine 5'-monophosphate during uptake by Streptomyces griseus. Unlabeled adenine brought about a (delayed) decrease in radioactivity that had been taken up by phosphate-limited resting cells of Streptomyces griseus from [14C]adenine-labeled adenosine 5'-monophosphate (AMP). Inorganic phosphate, on the other hand, stimulated adenine uptake from AMP, presumably by activating an energy-dependent active transport mechanism. Unlabeled phosphate rapidly diluted the uptake of radioactivity from [32P]AMP. Adenine inhibited uptake of [32P]AMP but not that of [32P]orthophosphate; adenine is thought to act by inhibiting the cleavage of AMP. The uptake of 32P and 14C from double-labeled AMP showed marked differences; 32P was taken up much faster into both cells and nucleic acids. These data indicate that uptake of AMP components takes place after extracellular dephosphorylation of the nucleotide.


Fractionation of inducible alkane hydroxylase activity in Pseudomonas putida and characterization of hydroxylase-negative plasmid mutations. The plasmid-determined inducible alkane hydroxylase of Pseudomonas putida resolved into particulate and soluble fractions. Spinach reductase and spinach ferredoxin could replace the soluble hydroxylase component. Two alkane hydroxylase mutants show in vitro complementation (S. Benson and J. Shapiro, J. Bacteriol., 123: 759-760, 1975): one, alk-7, lacks an active soluble component and the other, alk-181, lacks an active particulate component. Together with previous results on a particulate alcohol dehydrogenase enzyme (Benson and Shapiro, J. Bacteriol., 126: 794-798, 1976), these results allowed us to assay three plasmid-determined inducible activities: soluble alkane hydroxylase (alkA+), particulate alkane hydroxylase (alkB+), and particulate alcohol dehydrogenase (alkC+). Growth tests and in vitro complementation assays revealed three groups of plasmid mutations that block expression of alkane hydroxylase activity: alkA, which so far includes only the alk-7 mutation; alkB, which includes alk-181 and 11 other mutations; and a pleiotropic-negative class, which includes nine mutations that lead to loss of alkA+, alkB+, and alkC+ activities. Thus, the alk+ gene cluster found on IncP-2 plasmids contains at least four cistrons. We believe it is significant that two of these determined the presence of membrane proteins. The accompanying paper shows that these loci are part of a single regulon.


Regulatory mutations of the Pseudomonas plasmid alk regulon. Pseudomonas putida strains with plasmids carrying pleiotropic alk mutations gave rise to alkane-positive "revertants," which differ from wild type. Some had restricted ability to utilize alkane and primary alcohol growth substrates, and others could grow on undecane and dodecanol, which are not utilized by alk+ strains. These revertants showed altered responses to normal inducers of alkA+, alkB+, and alkC+ activities. Some revertants were constitutive for these activities. Constitutive mutants could also be isolated directly from wild type, but they appeared spontaneously at a frequency of less than 2 X 10(-8). Regulatory mutations of all three types, pleiotropic negative, altered inducer specificity, and constitutive, were tightly linked in transduction crosses with a polar alkB mutation. These results demonstrate that the IncP-2 plasmid alk gene cluster constitutes a regulon. They also permit the identification of at least one cistron whose gene product participates in inducer recognition and suggest that the alkABC regulon is not under simple repressor control.


Chemotaxis by Bdellovibrio bacteriovorus toward prey. A chemotaxis assay system that uses a modified Boyden chamber was characterized and used for measurements of chemotaxis by Bdellovibrio bacteriovorus strain UKi2 toward several bacterial species. Bacteria tested included both susceptible and nonsusceptible cells (Escherichia coli, Pseudomonas fluorescens, Bacillus megaterium, and B. bacteriovorus strains UKi2 and D). None was attractive to bdellovibrios when present at densities below 10(7) cells per ml. Chemotaxis toward E. coli was studied most extensively; under conditions that minimized effects of osmotic shock to the cells, E. coli and exudates from E. coli at densities as high as 10(8) cells per ml failed to elicit a chemotactic response. Cell-free filtrates from mixed cultures of bdellovibrios and E. coli neither attracted nor repelled bdellovibrios. The data indicate that bdellovibrios do not use chemotaxis to locate prey cells.


Intracellular, periodic structures in the gliding bacterium Myxococcus xanthus. Electron microscopic observations of thin sections of Myxococcus xanthus vegetative cells revealed the presence of cytoplasmic bundles of 4- to 5-nm-diameter filaments running longtitudinally below the cell membrane and terminating in association with the envelope near one pole. Part of each bundle demonstrated a herringbone-like periodicity (approximately 12-nm spacing). This structure was observed in cells from shake cultures and in gliding cells fixed by several methods. It is proposed that the structure may be attached to the envelope near both poles in gliding cells and that the motive force for motility may be provided by its contraction and relaxation. In one of four nongliding mutants examined, the periodicity was indistinct or lacking. In this mutant another structure, comprised of linearly arrayed beads, was observed in association with the filamentous bundle. Another structure, characterized by major, transverse bands (approximately 34 nm apart), occurred in patches that may traverse the diameter of the wild-type cells in which the structure was observed.


Cell wall and morphological changes induced by temperature shift in Bacillus subtilis cell wall mutants. Bacillus subtilis RUB1012 and RUB1013 have the following phenotype when grown at 45 degrees C: no growth on tryptose blood agar base, growth as clumps of spheres in broth culture, a slow autolysis rate, and a low proportion of teichoic acid to peptidoglycan. Revertants of strain RUB1012 (RUB2032, RUB2012, and RUB2042) that could grow on tryptose blood agar base were isolated. Each revertant had a different proportion of teichoic acid to peptidoglycan. The nanomoles of phosphorus per milligram of cell wall at the nonpermissive temperature were 141, 160, 236, and 541 for strain RUB1012 and revertants RUB2032, 2012, and 2042, respectively, as compared with 1,100 for the parent strain. With most bacteriophage tested, plating efficiency was related to the amount of glucosylated teichoic acid. Scanning electron microscopy was used to study strain RUB2032 during a shift from growth at 30 degrees C to growth at 45 degrees C. The change from rod to sphere began with the thickening of the cylindrical portion of the cell. Caps of the cells appeared to be immune to the thickening process. During growth, the cells became progressively shorter and thicker, and cell separation was inhibited. When cells of strain RUB2032 were shifted from growth at 45 degrees C to growth at 30 degrees C, accumulation of an amorphous material on the outer surfaces of the cells preceded the change from sphere to rod morphology. Cells remained clumped, with rods appearing at the periphery of the clumps. Analysis by DNA-mediated transformation and PBS1-mediated transduction indicated that strains RUB1012 and RUB1013 have multiple mutations mapping in the same region as other cell wall mutations.


High performance liquid chromatography of the hydroperoxides produced by lipoxygenases. Separation of 13-hydroperoxylinoleic acid or 13-hydroperoxylinolenic acid from linoleic acid or linolenic acid, respectively, was carried out easily and quickly by high performance liquid chromatography on porous polymer gel (TSK-Gel LS-140) using n-hexane/ethanol as an eluent. An eluent containing a large amount of n-hexane (96%) made possible the separation of 9- and 13-hydroperoxylinoleic acids. These methods were applicable for analyses of the products obtained by the incubation of soybean lipoxygenase-1 [linoleate: oxygen oxidoreductase, EC 1.13.11.12] with linoleic acid or 13-hydroperoxylinoleic acid.


Amino acid sequences of the aplpha and beta chains of adult hemoglobin of the tupai, Tupaia glis. Globin prepared from hemoglobin of adult tupai (Tupaia glis) was separated into alpha and beta polypeptide chains by CM-cellulose column chromatography. The S-aminoethylated alpha polypeptide chain and S-carboxymethylated beta polypeptide chain were each digested with trypsin, and the sequences of all the peptides thus obtained were established. The ordering of these tryptic peptides in the alpha and beta polypeptide chains was deduced from the homology of their primary structures with that of human adult hemoglobin. In this way the primary structures of the alpha and beta polypeptide chains of tupai hemoglobin were established; 27 amino acids in the alpha polypeptide chain and 26  in the beta chain differ from those in human adult hemoglobin.


Purification and specificity of carboxypeptidase from Streptomyces griseus K-1. A carboxypeptidase of St. griseus K-1 (CPase S) was found to possess the specificities of both mammalian pancreatic CPase A and B. Three adsorbents for affinity chromatography were prepared by coupling l-Leu, d-Leu, and d-Arg with CH-Sepharose 4B. d-Arg-CH-Sepharose and l-Leu-CH-Sepharose retained the purified CPase S but d-Leu-CH-Sepharose did not. The activities of CPase S toward CGL and BGA were eluted in the same position. CPase S migrated as a single band on polyacrylamide gel electrophoresis and the two activities were both extracted from this band on the gel.


Thioltrypsin. Chemical transformation of the active-site serine residue of Streptomyces griseus trypsin to a cysteine residue. The active-site serine residue of Streptomyces griseus trypsin was converted to a cysteine residue, and the product, thioltrypsin, was purified through two chromatographic steps with organomercurial-Sepharose and soybean trypsin inhibitor-Sepharose as specific adsorbents. The purified preparation of thioltrypsin was found to contain a single residue of cysteine and to react with almost equimolar amounts of normality titrants. It exhibited only traces of catalytic activity toward typical trypsin substrates such as Nalpha-tosyl-L-arginine methyl ester, whereas it retained some activity toward "active ester" substrates such as Nalpha-carbobenzoxy-L-lysine p-nitrophenyl ester. The activity was inhibited by sulfhydryl-blocking reagents, but no inhibition was observed by reagents reactive with the active hydroxyl group of serine proteases. Leupeptin, a natural trypsin inhibitor of peptidyl nature, also inhibited thioltrypsin. Some difference in the mode of leupeptin inhibition, however,  was detected between trypsin and thioltrypsin. The bindings of small synthetic ligands and soybean trypsin inhibitor to thioltrypsin were compared with those to trypsin.


Glutamine phosphoribosylpyrophosphate amidotransferase from Bacillus subtilis. A novel iron-sulfur protein. Glutamine phosphoribosylpyrophosphate amidotransferase, purifed to better than 98% purity from derepressed Bacillus subtilis, exists as a tetramer and as a dimer of apparently identical subunits with a molecular weight of 50,000 each. The enzyme contains 3 atoms of iron and 2 atoms of inorganic sulfide per subunit and has a yellow-brown color. The absorption spectrum is not altered by dithionite, but exposure to oxygen causes inactivation and partial bleaching of the visible spectrum. Thus, the Bacillus amidotransferase exhibits novel structural features and a new reaction type of proteins of the iron-sulfur group.


Hydroxylaminolysis of penicillin binding componenets is enzymatically catalyzed. The hydroxylaminolysis of the penicilloyl moiety from [14C]penicillin G binding component (PBC) complexes of the Bacillus subtilis D-alanine carboxypeptidase and of the mixture of PBC's of Staphylococcus aureus was inhibited by denaturation of the complexes by heat (55 degrees), detergent (1% sodium dodecyl sulfate), or trichloroacetic acid. The kinetics of inhibition by denaturation were comparable to those of the inhibition of [14C]penicillin G binding to the PBC's and of carboxypeptidase activity of the B. subtilis enzyme under identical denaturing conditions. These data establish that the hydroxylaminolysis is an enzymatically catalyzed process suggesting that penicillin G is bound to an enzymatically active site. Treatment of the denatured [14C]penicillin G-carboxypeptidase complex with sodium borohydride or at pH 12 resulted in the release of the penicilloyl moiety. These results are consistent with a carboxylic ester bond for the penicilloyl-PBC instead of a thiolester linkage as was initially presumed.


Membrane-bound estrone as substrate for microsomal UDP-glucuronyltransferase. Nonpolar substrates of microsomal UDP-glucuronyl-transferase partition between the hydrophobic phase of the microsomal membrane and the bulk aqueous phase in a suspension of microsomes in water. Partitioning of estrone into the membranes was measured in the studies presented and was extensive. Comparison of the rate of conjugation of estrone and the rate of its release from microsomes into the bulk aqueous phase showed that the pool of estrone within the membrane is the substate for UDP-glucuronyl-transferase. The rate of conjugation of estrone was 6-fold greater than the rate of release of estrone from the membrane into the aqueous phase. Several additional experiments showed that the rate of glucuronidation of estrone did not depend on the amount of estrone in the bulk aqueous phase. It is concluded that the microsomal membrane serves to concentrate nonpolar substrates of UDP-glucuronly-transferase. The phospholipid region of the microsomal membrane also may be a co-factor of UDP-glucuronyltransferase in the sense that binding of estrone to the membrane restricts its orientation in a manner that facilitates catalysis.


Mechanism of palmityl coenzyme A inhibition of liver glycogen synthase. Palmityl-CoA inhibits free liver glycogen synthase; the concentration required for half-maximum inhibition is 3 to 4 micrometer. Almost complete inhibition was observed at 50 micrometer. Palmityl-CoA inhibition is associated with dissociation of the tetrameric enzyme into monomers, and binding of palmityl-CoA to the monomers. Glycogen-bound enzyme is also inhibited by palmityl-CoA, resulting in dissociation of the enzyme into monomers and concomitant release of the enzyme from the primer glycogen. Palmityl-CoA inhibition of the enzyme is partially reversed by the glycogen synthase activator, glucose-6-P, whereas sodium lauryl sulfate-inhibited enzyme is not reactivated by glucose-6-P. Sodium lauryl sulfate inhibition results in the dissociation of the tetramer into the monomers. Bovine serum albumin and cyclodextrin can prevent palmityl-CoA inhibition only when they are added prior to palmityl-CoA addition. The possible physiological role of palmityl-CoA in glucose homeostasis is discussed.


The pathways of assimilation of 13NH4+ by the cyanobacterium, Anabaena cylindrica. The principal initial product of metabolism of 13N-labeled ammonium by Anabaena cylindrica grown with either NH4+ or N2 as nitrogen source is amide-labeled glutamine. The specific activity of glutamine synthetase is approximately half as great in NH4+-grown as in N2-grown filaments. After 1.5 min of exposure to 13NH4+, the ratio of 13N in glutamate to 13N in glutamine reaches a value of approximately 0.1 for N2- and 0.15 for NH4+-grown filaments, whereas after the same period of exposure to [13N]N2, that ratio has reached a value close to unity and is rising rapidly. During pulse-chase experiments, 13N is transferred from the amide group to glutamine into glutamate, and then apparently into the alpha-amino group of glutamine. Methionine sulfoximine, an inhibitor of glutamine synthetase, inhibits the formation of glutamine. In the presence of the inhibitor, direct formation of glutamate takes place, but accounts for only a few per cent of the normal rate of formation of that amino acid; and alanine is formed about as rapidly as glutamate. Azaserine reduces formation of [13N]glutamate approximately 100-fold, with relatively little effect on the formation of [13N]glutamine. Aminooxyacetate, an inhibitor of transaminase reactions blocks transfer of 13N to aspartate, citrulline, and arginine. We conclude, on the basis of these results and others in the literature, that the glutamine synthetase/glutamate synthase pathway mediates most of the initial metabolism of ammonium in A. cylindrica, and that glutamic acid dehydrogenase and alanine dehydrogenase have only a very minor role.


GTP stimulates and inhibits adenylate cyclase in fat cell membranes through distinct regulatory processes. GTP and hormones activate, synergistically, adenylate cyclase in purified plasma membranes from rat adipocytes. Addition of chelating reagents (EDTA or ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid) or thiol-reducing reagents (dithiothreitol or 2-mercaptoethanol) results in marked inhibition of enzyme activity without altering the synergistic stimulatory effects of GTP and hormones. The inhibitory effects of the reagents required the presence of GTP, indicating that inhibition involves a GTP-dependent process. This process is separate from the GTP-dependent process responsible for activation of the enzyme since it is selectively abolished by pretreatment of fat cell membranes with trypsin. It is suggested that inhibition and activation of fat cell adenylate cyclase by GTP occur through distinct regulatory processes.


Interaction of anions with iron-transferrin-chelate complexes. Preliminary evidence suggested that phosphate or borote destabilize iron-ovotransferrin-nitrilotriacetate complexes in the absence of added bicarbonate. The iron-ovotransferrin-EDTA complex was prepared in the absence of bicarbonate, and a number of anions, including phosphate, sulfate, and citrate, were found to perturb the visible absorbance (lambdamax = 490 nm) of this complex. Other anions, such as chloride, nitrate, and perchlorate, had little or no effect on the spectrum. Also, when bicarbonate was added to a solution of the iron-transferrin-EDTA complex (A515 = 0.45), within 2 min, the visible absorbance had decreased to A515 = 0.13. Slowly a new peak appeared (lambdamax = 470 nm), evidently the iron-transferrin-CO3 complex. When these spectral changes were monitored in detail, the lack of an isosbestic point indicated the existence of one or more intermediates in the conversion of iron-transferrin-EDTA complex to the iron-transferrin-CO3 complex. Experiments using ternary complexes containing either 59Fe or [14C]EDTA show that both iron and EDTA nearly completely dissociate from the protein (most likely concomitantly within 2 min after bicarbonate is added. These observations are best explained by a paradigm which includes anion binding to the apoprotein. It is clear that there is an intimate relationship between anions and the binding of iron chelates by transferrin.


Nucleotide sequence of an arginine transfer ribonucleic acid from bacteriophage T4. The nucleotide sequence of a phage T4-coded low molecular weight RNA, previously designated polyacrylamide gel band epsilon, has been determined. This RNA can be arranged in the cloverleaf configuration common to tRNAs, with an anticodon sequence, U-C-U, which corresponds to the arginine-specific codons A-G-A and A-G-G; it is therefore assumed to be an arginine tRNA. The complete nucleotide sequence of this RNA species is: pG-U-C-C-C-G-C-U-G-G-U-G-U-A-A-U-Gm2'-G-A-D-A-G-C-A-U-A-C-G-A-U-C-C-U-U-C-U-A-A-G-psi-U-U-G-C-G-G-U-C-C-U-G-G-T-psi-C-G-A-U-C-C-C-A-G-G-G-C-G-G-G-A-U-A-C-C-AOH. The nucleotide sequence was determined by analysis of RNA, uniformly labeled in vivo, according to the conventional techniques. In addition, RNA synthesized in vitro in the presence of alpha-32P-labeled nucleoside triphosphates was analyzed through the use of nearest neighbor sequencing techniques. Although a unique sequence could not be determined by this latter analysis, restrictions on the sequence imposed by nearest neighbor data and secondary structure common to tRNA molecules allowed prediction of the correct nucleotide sequence.


Preliminary crystallographic investigations of two phycobiliproteins. Single crystal x-ray diffraction investigations are in progress on two phycobiliproteins. C-phycocyanin from Anabaena variabilis crystallizes in space group P63 with a = b = 154 A and c = 40 A. The crystallographic asymmetric unit is (alphabeta)2 with a total molecular mass of 7.0-10(4) daltons. B-phycoerythrin from Porphyridium cruentum crystallizes in space group R3 with a = b = 189 A and c = 60 A. This molecule has the unusual molecular stoichiometry (alphabeta)6gamma and the crystallographic asymmetric unit is (alphabeta)2gamma1/3. This requires that the gamma chain undergo a perfect 3-fold disordering about the crystallographic 3 axis, i.e. the gamma chain must occupy three symmetry-equivalent positions, each with an occupancy of one-third.


Calcium content and distribution as a function of growth and transformation in the mouse 3T3 cell. Total Ca content and that fraction of Ca sensitive to removal by the chelator ethylene glycol-bis(beta-aminoethyl ether)N,N,N',N'-tetraacetate (EGTA) have been investigated in the mouse 3T3 cell as a function of growth stage, transformation with SV40 virus, and serum levels of the media. Cells were allowed to grow through several doublings in media containing (45)Ca. The cellular content of (45)Ca was used to access total cell Ca. That fraction of (45)Ca removed by EGTA was presumed to represent primarily surface-localized Ca. The data are expressed on a per cell volume basis to compensate for size differences as a function of growth stage and transformation. During exponential growth phase, the 3T3 cell contains 525pmol Ca/mul cell volume. Of this, approx. 457 pmol/mul is not removable by EGTA and, presumably, is cytoplasmically located. This value is in close agreement with previous studies on the HeLa cell (470 pmol Ca/mul cell water after the removal of the surface Ca). The low level of EGTA- removable Ca present in the 3T3 cell during early exponential growth (68 pmol Ca/mul cell volume) increases progressively with increasing cell density, and upon quiescence it is sevenfold greater. In contrast, SV40- transformed 3T3 cells growing exponentially possess total levels of Ca which are approximately two-thirds the levels of the normal 3T3 cell. However, their EGTA-sensitive Ca is not significantly different from that of exponentially growing, normal 3T3 cells. As the transformed cells continue to grow at high density, their total ca and their sensitivity to EGTA do not change, in contrast to the normal 3T3 cell. Thus, an increase in Ca associated with the cell surface appears to be correlated with growth inhibition. This has been investigated further by regulating growth of the normal and transformed cell with alterations in the serum level of the media. In 4 percent calf serum the normal cell is stopped from continued proliferation. Growth stoppage under these conditions is characterized by a nearly fourfold increase in EGTA-removable Ca, similar to the increase observed upon quiescence in depleted 10 percent serum. Similar treatment of the transformed cell does not reduce its growth rate, nor does it significantly alter Ca distribution. However, at 0.5 percent medium serum levels, the SV40 3T3 growth rate is substantially reduced and, under these conditions, EGTA-removable Ca increases twofold.


Nonidentity of ribosomal structural proteins in growing and starved Tetrahymena. We have examined the ribosomal structural proteins isolated from vegetatively growing Tetrahymena pyriformis and from cells that had been starved of all nutrients for 24 h. Reproducible, nonartifactual differences in protein complement, primarily associated with the large ribosomal subunit, were found. The kinetics of change in ribosomal protein complement were followed both in refed and in newly starved cells. Furthermore, attempts at correlating a certain protein "phenotype" with a particular functional state of the ribosome were made. It was concluded that the alterations seen could not be correlated with a specific stage in the normal ribosome cycle. We did show, however, that the change in protein complement could occur as a result of altering preexisting ribosomes. In addition, we showed that the change correlates with a decrease in growth rate rather than being caused by the starvation conditions themselves. Speculations as to the functional significance of the protein changes are presented.


Interaction of chemotactic factors with human macrophages. Induction of transmembrane potential changes. The electrophysiology of chemotactic factor interaction with cultured human macrophages was investigated with standard intracellular recording techniques. In initial studies, E. coli endotoxin-activated serum, added to cell cultures during intracellular recordings, caused membrane hyperpolarizations which were greater than 30 s in duration, 10-50 mV in amplitude, and associated with decreased membrane resistance. Control serum produced smaller hyperpolarizations lasting 10-20 s and 5-30 m V in amplitude. Endotoxin-activated human serum deficient in the third complement component (C3) did not produce hyperpolarizations unless the serum was reconstituted with C3 before activation. Fractionation of normal activated serum by molecular seive chromatography (G-75 Sephadex) indicated that only fractions that eluted with an estimated molecular weight of 12,500 produced membrane potential changes. The active material that was chemotactic for the macrophages was identified as the small molecular weight cleavage product of C5, C5a, by heat stability (30 min at 56 degrees C) and inactivation by goat antisera to human C5 but not C3. 17 percent of macrophages stimulated with C5a exhibited a biphasic response characterized by a small (2-6 mV), brief (1-10 s) depolarization associated with a decreased membrane resistance preceding the larger and prolonged hyperpolarizations. Magnesium-ethylene glycol bis[beta-aminoethyl ether]N,N'-tetraacetic acid (Mg [2.5 mM]-EGTA [5.0 mM]) blocked the C5a-evoked potential changes, whereas colchine (10(- 6)M) and cytochalasin B (3.0 mug/ml did not. Hydrocortisone sodium succinate (0.5 mg/ml) decreased the percentage of cells responding to C5a. In related studies, synthetic N-formyl methionyl peptide (f-met-leu-phe), which had chemotactic activity for cultured macrophages, produced similar membrane potential changes. Repeated exposure of macrophages to C5a or f- met-leu-phe resulted in desensitization to the same stimulus. Simultaneous photomicroscope and intracellular recording studies during macrophage stimulation with chemotactic factor demonstrated that the membrane potential changes preceded membrane spreading, ruffling, and pseudopod formation. These observations demonstrate that ion fluxes associated with membrane potential changes are early events in macrophage activation by chemotactic factors


Determination of oxyphenonium bromide in plasma and urine by means of ion-pair extraction, derivatization and gas chromatography-electron-capture detection. A sensitive and selective method for the determination of the quaternary ammonium compound oxyphenonium bromide (Antrenyl), a drug with strong anticholinergic properties, in human plasma and urine is described. The method is based on ion-pair extraction of the cation with perchlorate, a re-extraction according to ion-pair principles with tetrapentylammonium as the counter ion, hydrolysis to cyclohexylphenylglycolic acid, derivatization of this acid to its pentafluorobenzyl ester and determination of the ester by gas chromatography and electron-capture detection. Quantitation is possible down to 2 ng/ml of oxyphenonium bromide using 1 ml of plasma and down to 200 ng/ml using 0.1 ml of urine. The method described can also be applied to other anticholinergic drugs with an ester function.


Analysis of polychlorinated biphenyls by high-performance liquid chromatography and capillary gas-liquid chromatography. Seventy fractions were obtained by vacuum distillation of a PCB mixture containing 42% of chlorine. Four of these fractions were chosen, the composition of which covered the whole of the observed region, and analysed by means of high-performance liquid chromatography (HPLC) on silica gel using n-pentane as the mobile phase and capillary gas-liquid chromatography (GLC) using OV-101 and Carbowax 20M as stationary phases at 200 degrees C. The chosen distillation fractions were further prepared by HPLC, each yielding 10-14 samples. Individual PCB standards and all samples were analysed by HPLC and capillary GLC. These procedures permitted the identification of those compounds which are eluted simultaneously under the conditions used in either HPLC or capillary GLC alone.