Type I and II interferons and migration inhibitory factor: production in Mycobacterium bovis BCG-infected mice desensitized with old tuberculin or lipopolysaccharide.

Mice with delayed hypersensitivity induced by infection with Mycobacterium bovis strain BCG were desensitized by a single large dose of specific antigen (old tuberculin, OT) or a nonspecific interferon stimulus (bacterial lipopolysaccharide, LPS). Subsequent challenge of the desensitized animals revealed only a homologous hyporeactivity, that is, mice desensitized with OT showed decreased type II and migration inhibitory factor (MIF) responses to the specific antigen, which were unaffected by desensitization with LPS. Conversely, mice desensitized with LPS showed a decreased type I interferon and MIF response to LPS, which was unaffected by desensitization with OT. These results suggest that type I interferon and its accompanying low-titered MIF activity are produced by cell populations different from those that produce type II interferon and its accompanying high-titered MIF activity.


Platelet aggregation in rabbits made tolerant to endotoxin.

Endotoxin may cause abnormal deposition of platelet-endotoxin aggregates, and this event could have damaging effects. We compared the aggregation characteristics of platelets from rabbits made tolerant to the lethal effects of endotoxin with those of platelets from normal rabbits. Platelets from tolerant rabbits aggregated more rapidly (greater than 90 s faster) in the presence of endotoxin than did platelets from nontolerant animals. Furthermore, platelets from tolerant animals aggregated reversibly. These characteristics of platelets from tolerant animals are due to humoral factors in the plasma, because 1:1 dilution of normal platelet-rich plasma with plasma from tolerant rabbits caused the normal platelets to behave like those from tolerant animals. Survival after challenge with lethal quantities of endotoxin was enhanced in tolerant rabbits; this may be due to promotion of more efficient removal of endotoxin-platelet complexes from the blood by the reticuloendothelial system.


Kinetics of the humoral and cellular immune response of guinea pigs after injection of the synthetic adjuvant N-acetylmuramyl L-alanyl-D-isoglutamine: comparison with Freund complete adjuvant.

We studied the time course of humoral and cellular immunity of guinea pigs injected with the synthetic adjuvant N-acetylmuramyl L-alanyl-D-isoglutamine (MDP Pasteur, 10 microgram) in Freund incomplete adjuvant; the kinetics were compared with those obtained with Freund complete adjuvant (50 microgram of whole Mycobacterium butyricum). The antibody response to ovalbumin was faster and higher with MDP, but dropped sooner to a low level; the secondary response was, however, again higher for MDP than for Freund complete adjuvant. Cellular immunity, as measured by delayed hypersensitivity, and migration inhibition factor production werepositive for both adjuvants. The same response was followed in animals injected with MDPA, the nonamidated analog of MDP; the same kinetics as for Freund incomplete adjuvant were obtained for the primary response, but the secondary response was stronger and gave a positive delayed hypersensitivity reaction.


Biochemical examination of sera during systemic Candida infection in mice.

Candida pathogenesis was examined by intravenous challenge of mice with either C. albicans or C. guilliermondii. Animals were moribund 12 h postchallenge with C. albicans and were found to have the greatest number of organisms in the heart and kidney, severe interstitial myocarditis, and elevated serum levels of blood urea nitrogen, creatine phosphokinase, serum glutamic oxalacetic transaminase, serum glutamic pyruvic transaminase, and lactic dehydrogenase. In contrast, challenge with C. guilliermondii resulted in a significantly lower renal census, no myocarditis, and no significant change in the concentration of these serum constituents. Challenge with nonviable C. albicans did not produce the effects observed with viable organisms. Moreover, challenge with filamentous C. albicans resulted in biochemical alterations of lower magnitude and in lower mortality rates. These results indicated that altered serum biochemistries were correlated with the histopathology of fatal Candida infection and that there were distinct differences with C. guilliermondii and the dimorphic phases of C. albicans.


Oral hygiene and dental caries--a review.

Numerous studies have related dental caries experience of children with the frequency of their toothbrushing or with their oral cleanliness at the time of examination. Conflicting results were obtained and, therefore, no conclusions may be drawn from these studies. In other studies, the incidence of new carious lesions over a number of years was related to oral cleanliness during the same period of time. These studies showed that children with good oral hygiene had a lower caries incidence than those with poor oral hygiene, but the differences were small. Toothbrushing with a fluoridated toothpaste, which is now the usual practice, is effective in reducing caries incidence. Flossing can remove plaque from approximal tooth surfaces, and there is some evidence that it can reduce caries incidence. Dental caries can be largely prevented by highly efficient removal of plaque by toothbrushing and flossing. However, the available evidence suggests that regular prophylaxis by dental personnel is essential to maintain the necessary high level of efficiency. It is not known to what extent individuals are capable of removing their own plaque to the same high level of efficiency. It is probable that only highly-motivated individuals can maintain such a high standard of plaque control.


Effect of Trichinella spiralis infection on delayed hypersensitivity to heterologous antigens.

Studies were initiated to determine the effects of the different phases of the nematode life cycle on the delayed-type hypersensitivity (DTH) capabilities of the host. Mice were injected intraperitoneally with 4 X 10(6) viable or heat-killed BCG at 0, 3, 7, 14 and 28 days after oral infection with 200 Trichinella spiralis larvae. Groups were tested for DTH 28 days after BCG inoculation by the footpad swelling assay. Infection with T. spiralis was found to suppress the response in mice administered viable BCG 0 or 3 days later. Mice injected with viable bacteria 7 days after T. spiralis regained the ability to develop in vivo DTH reactions against old tuberculin. Groups administered viable BCG 14 and 28 days after nematode infection yielded potentiated DTH responses, when compared with control mice. Animals infected with T. spiralis and later injected with heat-killed BCG required a longer interval to develop specific immunopotentiation. Maximum immunopotentiation seemed to be related to the presence of larvae in muscle tissue (28 days). These findings suggest that the effect of T. spiralis on the immune capabilities of the host is dependent on the different anatomical locations of the parasite.


Host defense in cryptococcosis. I. An in vivo model for evaluating immune response.

An inbred mouse model was used to evaluate in vivo host immune response to Cryptococcus neoformans. Within 1 week of immunization, mice developed delayed type hypersensitivity reactions (DTH) to cryptococcal extracts derived from either culture filtrates or disrupted cells. There was no significant cross reactivity with extracts of other fungi. Previous immunization provided considerable protection against subsequent challenge with multiple strains of cryptococci. DTH also developed after nonimmunized mice were challenged with C. neoformans; however, in this situation DTH was not associated with prolonged survival. These studies indicate that mice can be immunized and protected against cryptococcosis and that this protection is associated with acquisition of DTH to cryptococcal antigens.


Alcohol and mood: an updated review.

Research since 1968 on the relationship between alcohol and mood is reviewed and updated. There are measurement and methodological barriers to delineating this relationship. The literature is replete with mixed findings that the motivation for drinking alcohol is psychological benefit, tension reduction, or affective improvement. Conflicting and equivocal data have characterized self-reports of depression and other affects in alcoholics. Evidence suggests that alcoholics experience increasing dysphoria as a consequence of alcohol consumption, while nonalcoholics anticipate--and generally attain--elevated moods as a result of drinking. Suggestions are made for further alcohol-mood research.


Locus of control research on alcoholic populations: a review. II. Relationship to other measures.

Research literature dealing with the relationships of locus of control to age, ability to function, and personality traits is reviewed. Results are contradictory on the relationship of locus of control in alcoholics to age, social desirability, and intellectual functioning. There is some tendency for internality to be related to better social functioning and the defenses of denial, intellectualization, and repression. There is some possible support for a relationship of externality and anxiety, and externality does appear related to helplessness, depression, isolation, general clinical pathology, and the defense of turning against another. No relationship has been found between locus of control and field dependence for alcoholics. Methodological difficulties have included problems with sampling, unsystematic research, assuming homogeneity of alcoholic samples, and assuming linearity and unidimensionality of the scales. Possible research which could clarify some of the areas are suggested.


Polio, politics, publicity, and duplicity: ethical aspects in the development of the Salk vaccine.

This paper is an historical account of the discovery, testing, and early distribution of the Salk polio vaccine. The discovery posed fundamental dilemmas of medical research, pharmaceutical production, and public health. This paper assesses the ethical problems which arose, and examines critically their resolution. The National Foundation for Infantile Paralysis (March of Dimes) financed and directed the discovery of the vaccine, subsequent field trials, and early distribution. The Foundation's role is analyzed with special attention to the conflicts between its philanthropic and scientific functions. The reat public demand which the discovery of the vaccine generated created a need for federal control which was only partly met. The federal government did not have sufficient institutional and legal mechanisms to assure the safety of the vaccine and protect the public. This discussion illustrates the failure of the government to keep pace with medical technology.


Computerized tomography of the orbit.

The potential for using CT to diagnose orbital lesions is clearly demonstrated in this chapter. The patients discussed were all suffering from proptosis and had other complaints such as visual disturbances, pain, or ophthalmoplegia. CT is superior to ultrasonography in its ability to reproduce anatomical structures, including the retroocular space, bony walls of the orbit, and extraorbital regions, such as ethmoid sinuses and the cranial cavity. Such reproduction helps distinguish lesions arising within the orbit from those invading the orbit from outside. CT not only defines the extent of a lesion but also provides information about the physical properties of the tissue. The remarkable difference on CT between proptosis caused by thyrotoxic disease and that caused by intraorbital tumor or pseudotumor is a striking example of the way in which CT may contribute to more accurate diagnosis.


Diagnostic ultrasonography.

Ultrasonography and the more recently developed technique of computerized tomography (see the preceding chapter) should be used concurrently in the evaluation and diagnosis of suspected orbital pathology. Ultrasonography is easily performed and safe; it has no known complications or contraindications. Unique information concerning the position and acoustical quality of tumors can be obtained. However, the interpretation of the ultrasonogram can be quite difficult; highly sophisticated equipment is needed to obtain the finer resolution required to make tissue diagnosis.


Abnormal ocular motility with brainstem and cerebellar disorders.

The disorders of ocular motility seen in association with brainstem or cerebellar disorders may point to rather specific anatomical or pathological correlations. Pontine gaze palsy reflects involvement of the pontine paramedian reticular formation. Internuclear ophthalmoplegia signifies a lesion in the medial longitudinal fasciculus. Skew deviation may result from a lesion anywhere in the posterior fossa. Ocular bobbing typically results from a pontine lesion. The Sylvian aqueduct syndrome is characteristic of involvement in the upper midbrain-pretectal region, usually a pinealoma. Cerebellar lesions may be manifested by gaze paresis, skew deviation, disturbances of saccadic or smooth pursuit movements, ocular myoclonus, or several characteristic forms of nystagmus. Familiarity with these disorders may be of great help to the physician dealing with a patient with a possible posterior fossa lesion.


The insensitivity of the chicken eye to the inflammatory effects of x-rays in contrast to its sensitivity to other inflammatory agents.

The effects of x-rays and three chemical agents, known to cause intraocular inflammation in mammalian eyes, were studied on the chicken eye because this species was reported to be insensitive to the cataractogenic effects of x-rays. Intravitreal injection of Shigella endotoxin and topical and/or intravitreal administration of PGE2, PGF2alpha, or arachidonic acid caused a breakdown of the blood-aqueous barrier, as indicated by flare and increased protein concentration in the aqueous humor. Following endotoxin injection, there was also a large accumulation of cells in the anterior chamber. The ocular inflammatory effects of endotoxin and arachidonic acid were inhibited by indomethacin. Thus the chicken eye reacts to these inflammatory agents in a manner similar to that previously described for the rabbit. In contrast, the inflammatory response which was reported to occur in the rabbit eye 3 to 4 hr after exposure to 500 or 1000 rads of x-rays was not observed in the chicken eye even after expsoure to 10,000 rads. Minimal flare and a small cellular infiltration were observed in some eyes only after extensive swelling of the surrounding tissues had developed. It is concluded that the insensitivity of the chicken eye to x-rays is due to some unique difference in the chain of events which mediates, or prevents, the effects of ionizing radiation rather than to a general insensivity to inflammatory agents.


Microsurgical anastomosis of vas deferens: an experimental study in the rat.

A patency rate of more than 90 per cent was achieved after 52 vasovasostomies. Satisfactory results were shown in both internally stented and nonstented anastomoses. A higher incidence of foreign body reaction was associated with silk sutures. Sperm granulomas and reduced patency were noted when superficial sutures were used indicating that a leakproof anastomosis is essential.


Desorption assay: a functional in vitro test for measuring the adhesion of E. coli on the urinary tract epithelium.

A functional test was developed to measure different adhesive properties of various Escherichia coli strains on the urinary tract epithelium. The test is based on initial incubation of everted canine ureteral segments in 2-hr cultures of E. coli (adsorption period), followed by sequential elutions in phosphate buffered saline. The release of bacteria (desorption rate) was measured from each elution by quantitative bacteriology. The function of the desorption assay was tested by showing different adhesive properties of various strains of E. coli. The strains isolated from infected canine urinary tracts showed high adhesion capacities superior to other strains of E. coli from different animal and/or tissue origin. This indicates a selective tissue and animal trophism determined by the strength of mucosal attachment. The test is also a valuable tool for measuring adhesion inhibiting substances in urine. The bulk of the adhesion inhibiting effect was localized in the urine ultrafiltrate.


[Immunity in syphilis].

Following a brief review of the characteristics of the causative organism of syphilis, its maintenance in laboratory animals and its relationship to the treponemes of the tropical treponematoses, consideration is given to the immune reactions and the development of antibodies to the treponeme of syphilis. The individual tests used in the diagnosis of syphilis, especially the complement fixation reactions and the flocculation tests, as well as the reactions employing specific antigens, such as the Treponemal Immobilisation Test (TPI), the Reiter Protein Complement Fixation Test (RPCFT), the Fluorescent Treponemal Antibody Test (FTA-ABS) and the Treponema Pallidum Haemagglutination Assay (TPHA), are discussed in detail. An evaluation of the results of the tests and the possible sources of errors are considered. The problem of the persistence of treponemes after clinically successful treatment is analysed and existing literature reviewed. The paper ends with a discussion of existing knowledge on protective immunity against infection with Treponema pallidum and a reminder of the need for further research into the immunology of syphilis.


[Psoriasis and bullous dermatoses].

In the last five years we observed in Berlin seven cases of psoriasis vulgaris combined with bullous eruptions of the pemphigoid type. Exogenious factors provoking the bullous eruptions were sun exposure, photochemotherapy and treatment with dithranol. These factors did not cause contact allergy of type IV, but pemphigoid eruption which could be characterised by immunfluorescence. This, was accompanied in two cases with high numbers of eosinophils in tissue and blood. The results show similarities with the findings of Person and Rogers who have shown the manifestation of otherwise subclinical pemphigoid in patients being treated for psoriasis.


Enzyme linked immunosorbent assay (ELISA) for determination of Staphylococcus aureus enterotoxin type B.

Determination of Staphylococcus aureus enterotoxin type B using ELISA is described. The ELISA is more sensitive than the micro-slide technique. The specificity of the determination of enterotoxin B depends on the specificity of the antisera used. Application of this new assay is evaluated and discussed.


Comparison of the Minitek test system with a conventional screening procedure for identification of Enterobacteriaceae.

A Minitek basic 9 disc/11 test system plus arabinose, inositol and rhamnose was compared with a conventional six tube screening system for identification of members of the family Enterobacteriaceae. Correlation varied from 88.3 to 100% with the various substrates when studied against 737 strains of these and other gram-negative bacteria recently isolated in our clinical laboratory. Two discs, citrate and ornithine, which originally gave a correlation of 88.3 and 95.5% respectively were modified so that a final correlation of 99.4 and 96.3% was obtained. The average correlation using the eleven discs was 97%.


Micromethod for rapid identification of gram-negative, nonfermentative bacteria.

Because of the contemporary significance of gram-negative, nonfermentative bacteria as etiological agents, a simple, rapid determinative system is essential. Therefore, an accurate, reproducible, and an inexpensive micromethod for the identification of these organisms has been developed. Included in this system are twenty-five biochemical tests. Carbohydrate utilization is demonstrated by modification of Otto and Pickett's formula for oxidative attack and assimilation of carbon sources, while the other substrates are modifications of commercially available products. Inoculation is a two-fold procedure into a plastic multicompartmented tray with wells containing 100 micro liters of each substrate. Initial inoculation yields 10(5) colony forming units per well. The carbohydrates are supplemented with an additional 50 micro liters of a 1 X 10(11) saline suspension of organisms. Reactions are read after a maximum incubation of 48 hr at 35 C. The results obtained with the identification of 124 strains of nonfermentative bacteria utilizing a conventional media schema as suggested by the Center for Disease Control and the micromethod system demonstrated the high reproducibility and correlation achievable with this rapid and economical microtechnique.


Patient participation in the patient-provider interaction: the effects of patient question asking on the quality of interaction, satisfaction and compliance.

The purpose of this study was to investigate the effectiveness, dynamics, and consequences of a health education intervention designed to increase patient question asking during the patient's medical visit. Data were collected at a Baltimore family and community health center which provides outpatient services to a low income, predominantly black and female population. The majority of the study participants were, in addition, elderly and chronically ill. A total of 294 patients and 3 providers took part in the study. The study design included random assignment of patients to experimental and placebo groups with two non-equivalent (non-randomized) control groups. Findings included: (1) The experimental group patients asked more direct questions and fewer indirect questions than did placebo group patients. (2) The experimental group patient-provider interaction was characterized by negative affect, anxiety, and anger, while the placebo group patient-provider interaction was characterized as mutually sympathetic. (3) The experimental group patients were less satisfied with care received in the clinic on the day of their visit than were placebo patients. (4) The experimental group patients demonstrated higher appointment-keeping ratios (an average number of appointments kept divided by an average number of appointments made) during a four-month prospective monitoring period.


The evaluation of continuing medical education: a literature review.

The literature of continuing medical education (CME) program evaluation was reviewed and analyzed. The "State-of-the-art" of CME evaluation is depicted in a series of tables which indicate evaluation methods and results. Evaluation designs providing strong assurances of valid results were not often found. The weakness of most published evaluations limit possible conclusions about the effectiveness of CME.


Lysozyme antigenicity and tissue fixation.

The preservation of lysozyme (LZM) antigenicity was studied in paraffin embedded tissue blocks. The reactivity for LZM varied with the type of tissue studied, the fixative used, the osmolarity and pH of the fixative, fixation time and temperature, and the method of dehydration. In both rat and human tissues aqueous fixatives were superior to nonaqueous fixatives in retaining LZM antigenicity. Brief fixation in fixatives of low osmolarity enhanced LZM staining in the parenchymatous tissues but diminished staining in human cartilage; prolonged fixation in fixatives of high osmolarity gave opposite results. Least affected by fixation was the LZM antigenicity in the serous cells of the glands of the respiratory tract. These cells also stained most intensely for LZM of all autopsy material studied.


[The therapy of tinnitus resulting from blast injury (author's transl)].

In a prospective study aimed at the cure of tinnitus due to blast injury, oral treatment of 172 patients with the mono-substances Betahistine, Pentoxifyllin and Xantinol-nicotinate were compared with those of control patients who received no medications. Patients treated with Pentoxifyllin and Xantinol-nicotinate improved better than the comparative group without therapy although the differences were not considerable. Betahistine as compared with the other groups produced significantly better therapeutic results.


[The management of mandibular fractures by internal fixation (author's transl)].

The results of treating mandibular fractures with rigid internal fixation are discussed. This method was used in 55 patients through the application of dynamic compression plates. The technique was then assessed critically and the followup treatment evaluated. Indications and contraindications for the procedure are discussed, as are possible errors or complications. In addition, selected cases illustrating the advantages of the procedure for reconstruction and functional rehabilitation of mandibular defects following ablative surgery or trauma are described.


DNA replication pattern and cell wall growth in Escherichia coli PAT 84.

An electron microscopic radioautographic study was made of tritiated thymidine incorporation into the genome of Escherichia coli PAT 84 and of tritiated meso-D,L-2,6-diaminopimelic acid (DAP) into the cell envelope. Pulse-labeled cells growing at 30 degrees C with a doubling time of 170 min were classified according to length by the method of agar filtration. Mathematical analysis of the length distribution led to the assumption of an exponential relation between length and time. A novel DNA replication pattern was found. Within the cell cycle DNA replication terminates at 70 min; then a gap follows of 64 min, after which DNA replication is initiated at 134 min. Thus, the C period is 106 min and the D period is 100 min. Cell constriction starts at 141 min and coincides with initiation of DNA replication. Detailed quantitative analysis of the [3H]thymidine grain frequency distribution allowed the distinction of three groups of cells. The first group incorporated no label, the second group an amount C, and the third group an amount 2 X C. The relative contribution of each group to a particular length class was determined. The data fitted very well into the DNA replication pattern. The same analysis was carried out on DAP pulse-labeled cells. Again, three groups of cells could be distinguished, and their relative contributions to each length class was determined. The group with the double amount of label was especially prominent at the end of the cell cycle. The emergence of this group might represent the acquisition of new lateral growth areas.


Effect of R-plasmid RP1 and nutrient depletion on the gross cellular composition of Escherichia coli and its resistance to some uncoupling phenols.

The resistance of Escherichia coli batch cultures depleted of carbon (C-dep), magnesium (Mg-dep), or phosphate (P-dep) against low concentrations of 3-chlorophenol, 4-chlorophenol, or 2-phenoxyethanol varied. C-dep cultures were always significantly more sensitive than Mg-dep or P-dep cultures. The presence of R-plasmid RP1 increased the sensitivity of C-dep cultures to 3- and 4-chlorophenol, yet had little effect on those cultured depleted in magnesium or phosphate ions. Cultures with R-plasmid RP1 had increased levels of beta-polyhydroxybutyrate irrespective of the nature of the depleting nutrient. P-dep bacteria had less than one-third of the phospholipid of other cell types, this deficiency being compensated for by increases in fatty acid and neutral lipid content. The reduction in phospholipid content of P-dep cultures was entirely accounted for by decreased diphosphatidylglycerol and phosphatidylethanolamine levels in these cells.


Evolution of ribosomal proteins in Enterobacteriaceae.

The evolution of ribosomal proteins of about 70 bacterial strains belonging to the family Enterobacteriaceae has been studied by use of previously reported data (S. Osawa, T. Itoh, and E. Otaka, J. Bacteriol. 107:168-178, 1971) and those obtained in this paper. The proximity of the bacteria was quantified by co-chromatographing the differentially labeled ribosomal proteins from two strains on a column of carboxymethyl cellulose in various combinations. The were then classified into 12 groups (=species?) according to their ribosomal protein compositions and were placed in a phylogenic tree.


Periplasmic localization of nicotinate phosphoribosyltransferase in Escherichia coli.

Nicotinate phosphoribosyltransferase (NAPRTase) in Escherichia coli mediates the formation of nicotinate mononucleotide, a direct precursor of nicotinamide adenine dinucleotide (NAD), from nicotinate and 5-phosphoribosyl-1-pyrophosphate. Specifically, NAPRTase contributes to NAD synthesis by utilizing intracellular nicotinate formed from NAD degradation products, which are recycled by NAD cycle enzymes and exogenous nicotinate when it is available. In previous studies, it has been tacitly assumed that almost all NAD cycle enzymes are localized in the cytoplasm of E. coli. The results of this investigation provide evidence that NAPRTase is a periplasmic (extracytoplasmic) enzyme. The osmotic shock of exponential-phase cells of E. coli K-12 and ML 308-225 resulted in the release of 63 to 72% and 42 to 48%, respectively, of the NAPRTase into the shock medium. In addition, when exponential cells of strains K-12 and ML 308-225 were converted into spheroplasts, 75 to 84% and 54 to 68%, respectively, of the enzyme was released into the spheroplast medium. Since previous estimates of the effective levels of NAPRTase present in putative repressed and derepressed E. coli cells appeared to be very low, a more convenient and accurate alternative method for the evaluation of NAPRTase in whole cells was developed. The results show that NAPRTase is subject only to a modest degree of enzyme repression. In addition, no evidence was found for the presence of a protein or low-molecular-weight inhibitor of the enzyme in repressed cells.


Role of glutathione in reversing the deleterious effects of a thiol-oxidizing agent in Escherichia coli.

Diamide was found to be much less specific for the oxidation of glutathione in vivo in Escherichia coli than had been previously assumed. In vivo, only a slight alteration of the ratio of reduced to oxidized glutathione was found, whereas a significant amount of glutathione was found in the form of mixed disulfide with proteins. This latter occurrence was postulated as being responsible for the bacteriostatic effect of diamide.


Intraperiplasmic growth of Bdellovibrio bacteriovorus on heat-treated Escherichia coli.

Heat treatment (55 degrees C for 40 min) of cell suspensions in buffer (ca. 3 x 10(9) cells per ml) of Escherichia coli ML35 caused a 4- to 4.5-log loss of cell viability. Similar results were found for several other E. coli strains that were examined. As a result of this heat treatment, 260-nm- and 280-nm-absorbing materials were released into the suspending buffer, along with about 10% of the total cellular radioactivity, when cells uniformly labeled with (14)C were used. In comparison with untreated cells, heat-treated E. coli ML35 cells showed (i) no significant changes in macromolecular composition other than ca. 22% less RNA content, (ii) an increased permeability to o-nitrophenyl-beta-d-galactopyranoside (a compound to which untreated cells are impermeable), (iii) almost complete loss of respiratory potential, and (iv) substantial losses of numerous glycolytic enzyme activities in cell extracts prepared from these cells. Intraperiplasmic development of Bdellovibrio bacteriovorus 109J with heat-treated E. coli ML35 as substrate cells appeared normal when observed microscopically, although bdellovibrio attachment and resultant bdelloplast formation were slightly retarded. No significant changes were observed in cell yields or in the ratios and contents of DNA, RNA, or protein between bdellovibrios harvested from untreated cells and those from heat-treated substrate cells after single-developmental-cycle growth on these cells. The average Y(ATP) values for intraperiplasmic growth on untreated and heat-treated substrate cells were 16.0 and 17.9, respectively. It is concluded that intraperiplasmic bdellovibrio growth on gently heat-treated E. coli substrate cells is very similar to growth on untreated substrate cells, even though the former substrate cells are nonviable and substantially impaired in many metabolic activities.


Outer membrane proteins of Escherichia coli. VII. Evidence that bacteriophage-directed protein 2 functions as a pore.

Protein 1, a major protein of the outer membrane of Escherichia coli, has been shown to be the pore allowing the passage of small hydrophilic solutes across the outer membrane. In E. coli K-12 protein 1 consists of two subspecies, 1a and 1b, whereas in E. coli B it consists of a single species which has an electrophoretic mobility similar to that of 1a. K-12 strains mutant at the ompB locus lack both proteins 1a and 1b and exhibit multiple transport defects, resistance to toxic metal ions, and tolerance to a number of colicins. Mutation at the tolF locus results in the loss of 1a, in less severe transport defects, and more limited colicin tolerance. Mutation at the par locus causes the loss of protein 1b, but no transport defects or colicin tolerance. Lysogeny of E. coli by phage PA-2 results in the production of a new major protein, protein 2. Lysogeny of K-12 ompB mutants resulted in dramatic reversal of the transport defects and restoration of the sensitivity to colicins E2 and E3 but not to other colicins. This was shown to be due to the production of protein 2, since lysogeny by phage mutants lacking the ability to elicit protein 2 production did not show this effect. Thus, protein 2 can function as an effective pore. ompB mutations in E. coli B also resulted in loss of protein 1 and similar multiple transport defects, but these were only partially reversed by phage lysogeny and the resulting production of protein 2. When the ompB region from E. coli B was moved by transduction into an E. coli K-12 background, only small amounts of proteins 1a and 1b were found in the outer membrane. These results indicate that genes governing the synthesis of outer membrane proteins may not function interchangeably between K-12 and B strains, indicating differences in regulation or biosynthesis of these proteins between these strains.


Isolation of an Escherichia coli K-12 dnaE mutation as a mutator.

Using a papillation method, a large number of Escherichia coli K-12 mutator mutations have been isolated. Only one of these (out of 1,250) mutator mutations has proved to be conditionally lethal at high temperatures. In vivo complementation tests indicated that this mutation, dnaE9, lies in dnaE, the structural gene for DNA polymerase III. The dnaE9 polymerase was not thermolabile in vitro; however, it showed a slow decline in specific activity in vivo at the nonpermissive temperature. Cultures of this mutant exhibited a comparably slow shutoff of DNA synthesis on shift to a nonpermissive temperature. dnaE9 showed temperature-sensitive mutator activity, which is not dependent on recA.


Isolation and characterization of mutants of the plasmid pKM101 deficient in their ability to enhance mutagenesis and repair.

A screening procedure was developed for identifying mutants of the plasmid pKM101 no longer capable of enhancing mutagenesis. The test was based on the large pKM101-mediated increase in the number of Gal+ papillae observed on colonies of Salmonella typhimurium gal mutants plated on tetrazolium-galactose plates in the presence of a mutagen. The pKM101 mutant plasmids transferred normally, were stably maintained in cells, caused normal levels of ampicillin resistance, and still imparted sensitivity to phage Ike to their hosts. However, the pKM101 mutants had lost the ability to (i) enhance the reversion of both point and frameshift mutations, (ii) protect the cells against killing by UV irradiation, (iii) increase the spontaneous reversion rates of point mutations, (iv) enhance plasmid-mediated reactivation of UV-irradiated phage P22, (v) enhance Weigle reactivation. One pKM101 mutant with different properties from the others was identified by its increased spontaneous mutator effect. It is suggested that pKM101 amplifies the activity of the inducible error-prone repair systems in bacteria and that this is the function of pKM101 in the Ames Salmonella tester strains used for detection of carcinogens as mutagens.


Increased antimetabolite sensitivity with variation of carbon source during growth.

In Serratia marcescens, analogs of leucine (norleucine), methionine (alpha-methylmethionine), histidine (3-amino-1,2,4-triazolealanine), tyrosine (p-aminophenylalanine), and tryptophan (7-methylindole) are conditional inhibitors of growth; inhibition occurs during the metabolism of some carbon sources but not with others. A further increase in sensitivity to growth inhibition by these analogs can be accomplished through the use of particular combinations of carbon sources present in the inoculum and in the subsequent analog-containing culture medium. Variable sensitivity to analog-mediated inhibition of growth observed during growth on glucose, glycerol, fructose, or citrate correlated inversely with the intracellular pool sizes of the amino acids cognate to the analogs used. The above-cited results, in conjunction with previous results obtained with Pseudomonas aeruginosa and Bacillus subtilis, involve diverse biochemical pathways and suggest that nutritional manipulation to alter the pattern of carbon flow in microorganisms is a generally useful means to accomplish increased sensitivity to growth inhibition by metabolite analogs.


Replication of plasmid DNA in Proteus mirabilis in limiting concentrations of thymine.

Replicating forms of the R plasmid pRR12 and the colicin E1 plasmid RSF2124 were isolated from Proteus mirabilis after growth in medium containing a limiting concentration of thymine. Both plasmids were replicated as partially supercoiled intermediates, which have densities between the values of covalently closed circular and nicked circular plasmid DNA in ethidium bromide-cesium chloride gradients. In addition, both plasmids had replication intermediates, which have densities lower than that of linear P. mirabilis chromosomal DNA. Some structural features of these replication intermediates were examined.


Pyridoxine-requiring mutants of Escherichia coli: glycolaldehyde dehydrogenase is not coded for by the pdxB gene.

Twenty-seven independent pyridoxineless mutants belonging to genetic linkage group I were assayed for glycolaldehyde dehydrogenase. Some mutants lacked enzyme activity entirely, and others showed activity ranging from very low to wild-type levels. Reversion to pyridoxine independence usually had no effect upon this activity. Transfer of the pyridoxine genes to a common host that had wild-type levels of enzyme activity made the recipient pyridoxineless without affecting the activity. These results negate the idea of an obligatory role for glycolaldehyde dehydrogenase in pyridoxine biosynthesis.


Analysis of cell division in single clones of the Escherichia coli K-12 lexA mutant.

The growth of clones of lexA mutant and lexA+ cells was analyzed. During normal growth lexA mutant clones frequently divided early, producing smaller newborn cells than the lexA+ clones. Some newborn cells in the lexA clones did not elongate or divide at all, a response that was never observed in the lexA+ clones. When starved for thymidine, most of the lexA mutant clones elongated and subsequently divided. The majority of lexA+ clones also elongated but did not divide. The above results suggest that one of the functions of the lexA+ gene is coordination of DNA repair with cell division.


Replication of colicin E1 plasmid DNA in vivo requires no plasmid-encoded proteins.

A derivative of bacteriophage lambda containing a colicin E1 plasmid replicon was constructed by recombinant DNA techniques. This phage, lambdacol100, has two functional modes of DNA replication; it can replicate via either plasmid or phage replication systems. lambdacol100 has been used to introduce the colicin E1 plasmid replicon into Escherichia coli previously treated with chloramphenicol to block protein synthesis. Under these conditions, lambdacol100 DNA is replicated normally as a colicin E1 plasmid. This suggests that colicin E1 plasmid replication in vivo does not require any plasmid-encoded proteins.


In vitro synthesis of cystathionine gamma-synthetase in Escherichia coli K-12.

Synthesis of cystathionine gamma-synthetase directed by DNA from a lambdadmet transducing phage has been achieved in cell extracts from Escherichia coli K-12. Enzyme synthesis was stimulated two- to threefold by the addition of guanosine 3'-diphosphate 5'-diphosphate to the incubation mixtures. Kinetic studies showed a 1.5- to 2.0-min lag between initiation of transcription and completion of a translatable message. This lag is shorter than that observed for beta-galactosidase synthesis with DNA from a lac transducing phage known to initiate transcription at the lac promoter. This result, together with information on the structure of the transducing phage, shows that pL is not used for initiation of in vitro metB transcription. Attempts to demonstrate repression were not successful, and unexpectedly, extracts from metJ+ strains were found to be more effective at enzyme synthesis than those from their metJ derivatives.


Permease-specific mutations in Salmonella typhimurium and Escherichia coli that release the glycerol, maltose, melibiose, and lactose transport systems from regulation by the phosphoenolpyruvate:sugar phosphotransferase system.

Several carbohydrate permease systems in Salmonella typhimurium and Escherichia coli are sensitive to regulation by the phosphoenolpyruvate:sugar phosphotransferase system. Mutant Salmonella strains were isolated in which individual transport systems had been rendered insensitive to regulation by sugar substrates of the phosphotransferase system. In one such strain, glycerol uptake was insensitive to regulation; in another, the maltose transport system was resistant to inhibition; and in a third, the regulatory mutation specifically rendered the melibiose permease insensitive to regulation. An analogous mutation in E. coli abolished inhibition of the transport of beta-galactosides via the lactose permease system. The mutations were mapped near the genes which code for the affected transport proteins. The regulatory mutations rendered utilization of the particular carbohydrates resistant to inhibition and synthesis of the corresponding catabolic enzymes partially insensitive to repressive control by sugar substrates of the phosphotransferase system. Studies of repression of beta-galactosidase synthesis in E. coli were conducted with both lactose and isopropyl beta-thiogalactoside as exogenous sources of inducer. Employing high concentrations of isopropyl beta-thiogalactoside, repression of beta-galactosidase synthesis was not altered by the lactose-specific transport regulation-resistant mutation. By contrast, the more severe repression observed with lactose as the exogenous source of inducer was partially abolished by this regulatory mutation. The results support the conclusions that several transport systems, including the lactose permease system, are subject to allosteric regulation and that inhibition of inducer uptake is a primary cause of the repression of catabolic enzyme synthesis.


Assignment of tra cistrons to EcoRI fragments of F sex factor DNA.

We describe here the cloning of single EcoRI fragments from the tra region of F DNA using ColE1::Tn3 as vector. These plasmids, as well as the series of Skurray et al. (Proc. Natl. Acad. Sci. U.S.A. 73:64-68, 1976), have been used to refine the map positions of tra cistrons on the F factor as well as to define a new DNA transfer cistron, traM. The current map of the tra cistrons is presented. None of the known tra cistrons, with the exception of traG, straddles an EcoRI site. The EcoRI site at 82 kilobases splits the traG cistron into two portions, an operator-proximal portion necessary for F pilus synthesis and an operator distal portion involved in conjugation itself. The operon structure of the tra cistrons was reevaluated, and we found that traI is at least partially independent of transcription of the traA to traD operon.


Transmembrane permeability channels in vesicles reconstituted from single species of porins from Salmonella typhimurium.

Aggregates of the "major" outer membrane proteins, "porins," of Salmonella typhimurium form diffusion channels in reconstituted vesicle membranes. The aggregate consists of three species of porins with apparent molecular weights of 34,000, 35,000, and 36,000 when active aggregates are subjected to sodium dodecyl sulfate-acrylamide gel electrophoresis after heating in the presence of sodium dodecyl sulfate (Nakae, J. Biol. Chem. 251:2176-2178, 1976). Single species of porins were isolated by solubilization of membranes and subsequent gel filtration in the presence of sodium dodecyl sulfate from the mutant strains of Salmonella typhimurium that produced only single species of porin. The single species of porins of either 34,000, 35,000, or 36,000 daltons formed diffusion channels when assayed for sucrose permeability in the vesicle membranes reconstituted from porins, phospholipids, and lipopolysaccharides. The exclusion limits of the pores made of single species of porins were not distinguishable from each other and from the exclusion limits of the pores made of the porin aggregates from the wild-type strain, when the permeability of vesicle membranes to radioactive di-, tri-, and tetrasaccharides and to various sizes of radioactive polyethylene glycol was determined. Porin-deficient mutants produced residual amounts of porin amounting to 1 to 5% that produced by the parent strain. This residual porin made diffusion channels when the isolated porins were incorporated into the vesicle membrane and assayed for permeability of saccharides.


Proteins synthesized in minicells containing plasmid ColE1 and its mutants.

Minicells containing the ColE1 plasmid and derivatives of it that contain a Tn3 insertion and deletions of the plasmid were shown to synthesize a variety of polypeptides: (i) a 56,000-dalton polypeptide was found that is colicin E1. (ii) A 15,000-dalton polypeptide is involved in plasmid DNA relaxation. (iii) A 14,000-dalton polypeptide may be the colicin immunity protein. (iv) A correlation between the location of various Tn3 insertions and the proteins synthesized from the plasmid DNA was found that suggests the locations of promoter sites in the plasmid and directions of transcription in two regions of the plasmid. (v) Polypeptides associated with the presence of Tn3 were identifiable. (vi) The nature and extent of gene expression of deletion mutants only generally correlates with the extent of the deletions.


Positive detection of mycoplasma contamination by the whole-mount preparation of cell cultures for transmission electron microscopy.

Low-level mycoplasma contamination of cell cultures is difficult to recognize with presently available techniques. This report describes the adaptation of the whole-mount technique, usually used for scanning microscopy, for transmission electron microscopy. The differentiation between microvilli and the equal-sized filamentous mycoplasma is based on the differential density obtained by the use of the method described. This method allows positive identification of mycoplasma and reduces the preparation time and the time necessary for scanning the preparation.


Translation of the leader region of the Escherichia coli tryptophan operon.

When the trp operon of Escherichia coli contains either of two deletions that fuse the initial portion of the leader region to the distal segments of the trpE gene, novel fusion polypeptides are produced. The new polypeptides are synthesized efficiently both in vivo and in vitro, and their synthesis is subject to repression by trp repressor. Fingerprint analyses of tryptic and chymotryptic digests of the new polypeptides show that both contain trpE polypeptide sequences and, despite their different sizes, share the same N-terminal sequence. Our results suggest that synthesis of the new polypeptides is initiated at the AUG-centered ribosome-binding site in the leader region and proceeds in phase to the region coding for the C-terminal end of the trpE polypeptide.


Role of murein lipoprotein in morphogenesis of the bacterial division septum: phenotypic similarity of lkyD and lpo mutants.

Phenotypes were compared in two different classes of mutants with defects in murein-lipoprotein (lkyD mutants of Salmonella typhimurium and an lpo mutant of Escherichia coli). Both mutations are associated with the same triad of phenotypic abnormalities, consisting of defective formation of the division septum, leakage of periplasmic proteins during growth, and increased sensitivity to several unrelated external toxic agents. The abnormality in septum formation consists of a defect in invagination of the outer membrane during formation of the nascent septum. The results suggest that formation of the murein-lipoprotein link plays an important role in differentiation of the division septum and perhaps also in maintaining the normal barrier function of the outer membrane.


Chitin synthesis in Candida albicans: comparison of yeast and hyphal forms.

Chitin synthesis was studied in both yeast and hyphae of the dimorphic fungus Candida albicans. Incorporation of N-acetyl-d-[1-(3)H]glucosamine ([(3)H]GluNAc) into an acid-alkali-insoluble fraction was 10 times greater in hyphal-phase cells. A crude preparation of chitin synthetase was obtained from sonically treated protoplasts of both forms of Candida. Enzyme activity, which was determined by using [(14)C]UDP-GLuNAc as a substrate, was exclusively associated with the 80,000 x g pellet from sonically treated protoplasts of both forms. It was determined that enzyme activity (nanomoles of [(14)C]UDP-GluNAc incorporated per milligram of protein) was approximately 2 times greater in hyphae versus yeast cells. Enzyme activity in both yeast and hyphae increased six- to sevenfold when the enzyme preparations were preincubated with trypsin. A vacuolar fraction, obtained from yeast cells but not from hyphae, stimulated enzyme activity when incubated with either yeast or hyphal enzyme preparations. Membrane fractions from protoplasts coated with [(3)H]concanavalin A before disruption were isolated by Renografin density gradient centrifugation. Chitin synthetase activity was preferentially associated with the concanavalin A-labeled fraction, suggesting that the enzyme was located on the plasma membrane. In addition, enzyme activity in protoplasts treated with cold glutaraldehyde before disruption was significantly greater than in protoplasts that were sonically disrupted and then treated with cold glutaraldehyde, indicating that the enzyme resides on the inner side of the plasma membrane.


New major outer membrane proteins found in an Escherichia coli tolF mutant resistant to bacteriophage TuIb.

Cell envelopes prepared from an Escherichia coli tolF strain selected as resistant to phage TuIb contained a new major outer membrane protein related to outer membrane proteins Ia and Ib. The strain that produces this protein is a tolF par double mutant but contains an additional mutation leading to the production of the new major outer membrane protein. Antibiotic sensitivity lost as a result of the tolF mutation is regained in strains that contain the new major outer membrane protein. This indicates that this protein functions to restore the selective permeability of the outer membrane to low-molecular-weight hydrophilic molecules.


Reaction order of Saccharomyces cerevisiae alpha-factor-mediated cell cycle arrest and mating inhibition.

Alpha-factor-mediated cell cycle arrest and mating inhibition of a mating-type cells of Saccharomyces cerevisiae have been examined in liquid cultures. Cell cycle arrest may be monitored unambiguously by the appearance of morphologically abnormal cells after administration of alpha factor, whereas mating inhibition is determined by comparing the mating efficiency in the absence or presence of added alpha factor. For both cell cycle arrest and mating inhibition, a dose-dependent response may be observed at limiting concentrations of the pheromone. If cell cycle arrest and mating inhibition require a small number of alpha-factor molecules, one might expect that responsive/nonresponsive cells = K(alpha factor)(N) where N is the order of dependence of cell cycle arrest (or mating inhibition) on alpha-factor concentration. The value of N has been determined to be 0.98 +/- 0.18 (standard error of the mean) for cell cycle arrest and 1.08 +/- 0.32 for mating inhibition. These results support the notion that saturation of a single site by alpha factor is sufficient to cause cell cycle arrest or mating inhibition of a mating-type cells.


Method for the genetic labeling of cryptic plasmids.

A recently developed method for detecting transposition was employed to genetically "label" conjugative plasmids such as F and Ent P307, which do not normally exhibit a readily identifiable phenotype.


Mutations affecting the citrate-dependent iron uptake system in Escherichia coli.

Isolation of six strains of Escherichia coli K-12 carrying mutations affecting the citrate-dependent iron uptake system is described. Genetic analysis of these mutants showed that the mutation affecting the citrate system are cluster together at about min 6 on the E. coli chromosome.


Phospholipid turnover during the division cycle of Escherichia coli.

The turnover of phospholipids in Escherichia coli B/r was analyzed in synchronously growing populations. The turnover of presynthesized phosphatidyl-glycerol and cardiolipin continued at a constant exponential rate throughout the division cycle.


Psychiatric illness and human renal transplantation.

This study focuses on physiological as well as psychological causes for psychiatric illness in renal transplant recipients. During an 18 month period 20 transplant recipients were evaluated in psychiatric consultation. A variety of psychiatric illnesses were noted with eight having a secondary depression diagnosed according to psychiatric research criteria. Case histories of these patients indicated that corticosteroid and meythyldopa therapy were significant etiologic agents in the development of the depressive syndrome. A step-wise approach to the management of patients with depressed mood or slowed thinking was proposed.


Ribonuclease III is involved in motility of Escherichia coli.

Mutants of Escherichia coli deficient in ribonuclease III are nonmotile. All transductants and revertants that regained ribonuclease III also regained motility, and all transductants that remained or became rnc are nonmotile, although only some of the revertants that regained motility also became ribonuclease III+.


Studies on the oligomeric structure of yeast aldehyde dehydrogenase by cross-linking with bifunctional reagents.

The molecular w:ight of yeast aldehyde dehydrogenase determined by sucrose density gradient centrifugation was 207,000 +/- 13,000. The enzyme activity was proportional to the enzyme concentration in the range of 2 X 10(-11) M to 1 X 10(-7) M. Cross-linking patterns obtained with yeast aldehyde dehydrogenase after treatment with a series of diimidoesters of increasing chain lengths with different reaction times resulted in the appearance of tetramers as the largest cross-linked product of the enzyme subunits. The molecular weights of its monomer, dimer, trimer, and tetramer were, 57,000, 114,000, 171,000, and 228,000, respectively, as estimated from their mobilities on SDS-electrophoresis. In tetramers monomers are probably assembled in a heterologous square arrangement.


Ergosterol biosynthesis in yeast. Pathways in the late stages and their variation under various conditions.

[Methyl-14C]methionine was supplied to yeast cells under aerobic and anaerobic conditions for the investigation of the pathway for ergosterol biosynthesis after the methylation of the side-chain. Under aerobic conditions, the incorporation of radioactivity into ergosterol was high. With a limited oxygen supply, in contrast, the radioactivity was first accumulated in ergosta-7,24(28)-dien-3beta-ol and ergosta-8,24(28)-dien-3beta-ol, and then transferred to ergost-7-en-3beta-ol, ergost-8-en-3beta-ol and ergosta-7,22-dien-3beta-ol with time. Under strictly anaerobic conditions, a double bond was introduced neither to delta5 nor to delta22. The results of the tracer experiments suggested the operation of several pathways in the late stages of ergosterol biosynthesis. It was also suggested that the main pathways varied depending on the conditions such as oxygen supply and other factors. The above conclusion was supported by the results of the analyses of the sterol compositions of the cells grown under various conditions.


Binding and functional properties of concanavalin A and its derivatives. II. A proteolytic product with saccharide-binding activity.

Limited digestion of the intact subunit of concanavalin A (Mr = 26,000) with trypsin followed by affinity chromatography on Sephadex G-100 has yielded a highly purified product designated here as Tn-Con A. Chemical studies have shown that Tn-Con A is composed of several components: a large fragment (Tn I, Mr = 19,000) spanning residues 1 to 172, and lower molecular weight polypeptides that are noncovalently associated with Tn I to form the active molecule. The molecular weight of Tn-Con A at pH 7 was 90,000, suggesting that, like native concanavalin A, it was a tetramer at physiological pH. Equilibrium dialysis experiments showed that Tn-Con A bound 1 molecule of alpha-methyl-D-glucoside/22,000 g atoms of protein and therefore that four saccharides are bound by the tetrameric molecule. Tn-Con A and native concanavalin A competed for the same receptors on the lymphocyte surface. Moreover, Tn-Con A was mitogenic for both mouse and human lymphocytes with dose-response curves similar to those of the native lectin. All of these results indicate that tryptic hydrolysis of concanavalin A produces a fragmented molecule retaining the saccharide-binding, subunit association, and mitogenic capacity of the native protein.


A multienzyme system for priming the replication of phiX174 viral DNA.

Synthesis of the oligonucleotides that prime replication of phiX174 single-stranded DNA employs complex protein machinery of the host cell which is probably used by the cell to replicate its own chromosome. Primer synthesis depends on at least five proteins (DNA binding protein, dnaB and dnaC proteins, protein i, and protein n) and ATP to form a replication intermediate and another protein, primase (dnaG protein), to assemble the oligonucleotide by template transcription. The data in this paper show that ribo- and deoxyribonucleoside triphosphates can serve as substrates and form hybrid primers when present together. Both RNA and DNA primers were initiated with ATP. At least three of the four base-pairing nucleoside triphosphates were required for the transcription that generates effective primers. Over 90% of the RNA and DNA transcripts were extended into complementary strands by DNA polymerase III holoenzyme. At optimal triphosphate concentrations, the rate and extent of primer formation were greater from ribonucleoside triphosphates than from deoxyribonucleoside triphosphates. Uncoupled from DNA replication, the length of RNA primers was 14 to 50 residues, the DNA primers 4 to 20 residues. The fingerprint pattern of an RNase digest of RNA primers has a complexity suggestive of transcription from many sites on the phiX174 template. The multienzyme priming system is highly specific for phiX174 DNA as template.


Urologic complications in renal homotransplantation.

Nineteen urologic complications developed in 161 renal transplant patients, 14 of whom developed cutaneous fistulae. The majority of complications occurred at the ureterovesical and ureteropelvic areas. Pelvic and vesical abnormalities were uncommon. Twelve patients with cutaneous fistulae were treated conservatively. The mortality rate in our series was 47%.


Excretion of lipoteichoic acid by group A streptococci. Influence of penicillin on excretion and loss of ability to adhere to human oral mucosal cells.

Group A streptococci were grown in the presence of [2-(3)H]glycerol. Concentrated suspensions of the labeled organisms were incubated with and without penicillin. [(3)H]Glycerol-labeled material accumulated in the supernates in increasing amounts with increasing concentrations of penicillin, ranging from 0 to 50 U/ml. The excretion of labeled material occurred in the absence of nucleic acid synthesis or bacteriolysis indicating that the phenomenon is independent of cell multiplication or decay. The accumulation of label was paralleled by an accumulation of erythrocyte-sensitizing material measured by passive hemagglutination tests for lipoteichoic acid antigen, indicating that a portion of the labeled material possessed the properties of lipoteichoic acid. Culture supernates were fractionated by column chromatography, and the materials obtained were analyzed by electrophoresis on sodium dodecyl sulfate polyacrylamide, thin-layer chromatography, and paper chromatography. The ability of the same materials to bind to human erythrocytes and epithelial cells was tested. The culture supernate contained lipoteichoic acid, deacylated lipoteichoic acid, glycerol phosphate, and free glycerol. Penicillin caused an increase in the amounts of each of the excreted materials. Streptococci that were stimulated with penicillin to lose their lipoteichoic acid (previously shown to mediate adherence of group A streptococci) lost their ability to adhere to buccal mucosal cells, suggesting that penicillin may influence bacterial ecology by mechanisms other than killing sensitive organisms.


Cardiac and renal prostaglandin I2. Biosynthesis and biological effects in isolated perfused rabbit tissues.

Both the isolated perfused rabbit heart and kidney are capable of synthesizing prostaglandin (PG) I(2). The evidence that supports this finding includes: (a) radiochemical identification of the stable end-product of PGI(2), 6-keto-PGF(1alpha), in the venous effluent after arachidonic acid administration; (b) biological identification of the labile product in the venous effluents which causes relaxation of the bovine coronary artery assay tissue and inhibition of platelet aggregation; and (c) confirmation that arachidonic acid and its endoperoxide PGH(2), but not dihomo-gamma-linolenic acid and its endoperoxide PGH(1), serve as the precursor for the coronary vasodilator and the inhibitor of platelet aggregation. The rabbit heart and kidney are both capable of converting exogenous arachidonate into PGI(2) but the normal perfused rabbit kidney apparently primarily converts endogenous arachidonate (e.g., generated by stimulation with bradykinin, angiotensin, ATP, or ischemia) into PGE(2); while the heart converts endogenous arachidonate primarily into PGI(2). Indomethacin inhibition of the cyclo-oxygenase unmasks the continuous basal synthesis of PGI(2) by the heart, and of PGE(2) by the kidney. Cardiac PGI(2) administration causes a sharp transient reduction in coronary perfusion pressure, whereas the intracardiac injection of the PGH(2) causes an increase in coronary resistance without apparent cardiac conversion to PGI(2). The perfused heart rapidly degrades most of the exogenous endoperoxide probably into PGE(2), while exogenous PGI(2) traverses the heart without being metabolized. The coronary vasoconstriction produced by PGH(2) in the normal perfused rabbit heart suggests that the endoperoxide did not reach the PGI(2) synthetase, whereas the more lipid soluble precursor arachidonic acid (exogenous or endogenous) penetrated to the cyclooxygenase, which apparently is tightly coupled to the PGI(2) synthetase.


Smoking treatment strategies, expectancy outcomes, and credibility in attention-placebo control conditions.

Assessed credibility of the rapid smoking procedure, covert sensitization a combined approach, satiation, and a relaxation technique in a group of 38 self-reported cigarette smokers. The results suggested that while sex is not a factor in credibility ratings, different treatment strategies do elicit varying degrees of confidence in terms of positive treatment expectances. Scheffé's method of posttest comparisons revealed that satiation was least preferred, while the relaxation procedure was rated the highest in credibility. The other aversive treatment strategies did not differ significantly from either the relaxation or satiation procedures. These findings are discussed as they relate to credibility as a nonspecific treatment variable that smoking researchers may effectively manipulate through a rating procedure in creating a more sound experimental design in attention-placebo control conditions.


Differential mood changes in alcoholics as a function of anxiety management strategies.

A study was designed to compare the effects of three relaxation strategies on mood state profiles in a group of 30 male alcoholics. The Profile of Mood States (POMS) was used for pre- and posttest treatment measures. The results indicated that the three relaxation strategies--progressive relaxation training, (PRT), meditation training (MT), quiet rest (QR)--produced qualitatively different patterns. While both PRT and MT produced measurable decrements in self-reported tension and QR controls showed no change on any of the six subscales, PRT was observed to effect decreased depression as well as a trend toward increased vigor. Meditation training effects were limited to decreased tension and a nonsignificant decrease in fatigue. These results are discussed in terms of refinement of the "relaxation response" notion to a more heterogeneous, technique-centered definition.


The effect of test taking on acceptance of bogus personality statements.

Ss received either positively or negatively toned feedback after taking a projective test. Other Ss merely received the  feedback statements without taking the test. All Ss rated the feedback statements individually and as a unit as to how well they described their personality and the personality of people in general. Feedback as a unit was more acceptable when (a) the test was taken, and (b) when positively toned feedback was received. Collapsing across feedback conditions, personal acceptability was higher for those who took the test than for those who did not take the test. The method allowed for analysis of individual items and thereby provided an additional measure of acceptability than simply using the overall acceptability score.


Outbreak of infantile enteritis caused by enterotoxigenic Escherichia coli O6.H16.

Ten out of 18 babies at risk developed enteritis in an outbreak in a special-care baby unit. Salmonella, Shigella, and Escherichia coli belonging to the traditional infantile enteropathogenic serogroups were not found in faeces from the babies and the staff, and no virus particles were found by electron microscopy. Detailed serotyping of E. coli showed that five of the 10 babies with diarrhoea and one of the eight without diarrhoea were excreting E. coli O6.H16. All six isolates of this serotype produced both heat-labile and heat-stable enterotoxin. Enterotoxigenic strains of E. coli O6.H16 have caused outbreaks of enteritis in adults in the USA and Japan.


Significance of alpha-subunit HCG demonstrated in breast carcinomas by the immunoperoxidase technique.

Fifty-three breast carcinomas were examined by an indirect immunoperoxidase technique for the presence of the alpha-subunit of human chorionic gonadotrophin. Positive staining occurred in 12 (22.6%). There was no correlation between alpha-subunit production and specific histological features, but production was related to the presence of lymph node metastases and hence a poorer prognosis. The alpha-subunit could be used as a prognostic indicator in the assessment of breast carcinoma.


Distribution of transferrin, ferritin, and lactoferrin in human tissues.

An immunoperoxidase staining technique was used for detecting three major iron-binding proteins (transferrin, ferritin, and lactoferrin) in routine histological paraffin sections of human tissue. Transferrin was found mainly in hepatocytes, a variety of epithelial and myoepithelial cells, renal tubular cells, and histiocytes. Ferritin was most readily found in histiocytes and liver cells, with weaker reactions seen in epithelial cells. Lactoferrin was found in lactating breast tissue, bronchial glands, polymorphs, and gastric and duodenal epithelial cells. The technique is potentially valuable for investigating abnormal iron states.


Ethanol formed from arabinose: a rapid method for detecting Escherichia coli.

Head-space gas-liquid chromatography showed that Escherichia col are consistent in their ability to ferment arabinose with the production of ethanol. In both still and shaken cultures ethanol production is closely aligned with growth. Also ethanol production is related to inoculum size so that varying sizes of inocula may be distinguished by choosing a time of sampling early in growth. The method is suitable as a rapid test for E. coli and is applicable for use in detecting E. coli urinary tract infections.


Head-space gas liquid chromatography for rapid detection of Escherichia coli and Proteus mirabilis in urine.

Measuring ethanol formed from arabinose by Escherichia coli and dimethyl disulphide formed from methionine by Proteus mirabilis is a rapid way of detecting these organisms in the urine. Ethanol and dimethyl disulphide are detected by head-space gas-liquid chromatography. Since sedimented organisms and not whole urine are used the method is not subject to interference from compounds within the urine. When it was tested on 122 samples of urine, 94 from patients with suspected urinary infection and 28 supposedly uninfected control specimens, the results correlated well with viable counts by a conventional bacteriological method. As well as being rapid (results available in four hours) the method is reliable, easy to use, and could be automated. It now requires extensive testing in a hospital laboratory.


Retinal ganglion cells in the crucian carp (Carassius carassius). II. Overlap, shape and tangential orientation of dendritic trees.

Ganglion cells were studied in methylene blue stained flat-mounted retinas. Three categories of cells are described: small (S) and large (L) ganglion cells in the main ganglion cell layer, and large ganglion cells (LD) with somata more or less displaced into the inner plexiform layer. These LD cells have two to four very thick primary dendrites and are identifiable as ganglion cells by their axons. An analysis of published data reveals that the large ganglion cells of the crucian carp (type L and LD) have several striking characteristics in common with the large ganglion cells of the dogfish, the frog and the cat: (1) they are selectively stained by methylene blue; (2) they comprise only 2-5% of all the ganglion cells; (3) the large cells can be divided into two or three subtypes, and within each subtype the dendritic trees usually cover the retinal surface with a two- or threefold overlap.  New ganglion cells are formed from neuroblasts at the retinal margin and most dendrites first grow along this neuroblastic zone. Thus the main dendrites of the L and LD cells tend to be oriented parallel to the margin all around the periphery of a crucian carp retina. Independent of the size of the eye this parallel orientation disappears at the same relative distance from the margin (about one-third of the distance from the margin to the optic disc). If all L and LD cells are formed at the retinal margin and first develop oriented dendrites, we have to assume that the more randomly oriented dendritic trees in the central retina have undergone a reorganization.


Effects of halofenate on glucose tolerance in patients with hyperlipoproteinemia.

Halofenate, a triglyceride- and uric acid-lowering drug, potentiated the effect of oral hypoglycemics. Its effect on serial glucose tolerance was evaluated in ten patients with hypertriglyceridemia without overt diabetes. Six-hour oral glucose tolerance tests were done during a control period and every 24 weeks over two years of halofenate treatment. Abnormal glucose tolerance (chemical diabetes) was observed during the control period in six of ten patients. The number of abnormal tests gradually decreased to none by 48 weeks. Plasma glucose, insulin, and free fatty acid values during the glucose tolerance tests were reduced significantly. Halofenate induced significant serum uric acid reduction. No significant regressions were observed among levels of lipids, hormones, glucose, and uric acid. The mechanisms by which lipid-lowering drugs improve glucose tolerance are as yet unexplained.


The forms of non-pyramidal neurons in the visual cortex of the rat.

Rapid Golgi preparations from area 17 of young adult rats have been studied to determine the morphology and distribution of non-pyramidal neurons. Such cells were observed in all of the cellular laminae of the cortex, but were particularly prevalent in layers IV and V. Non-pyramidal neurons were categorized according to two features: (1) dendritic projection pattern, and (2) abundance of dendritic spines. Dendritic patterns were classified as multipolar, bitufted, and bipolar, and spine patterns as spinous, sparsely spinous, and spine-free. Spinous dendrites were associated only with multipolar neurons, while sparsely spinous and spine-free dendrites were each associated with cells of all three non-pyramidal dendritic patterns. The most frequently observed non-pyramidal cell types were multipolar cells of the spine-free and sparsely spinous varieties. All of the general cell types encountered have been described in the literature on non-pyramidal neurons, indicating the lack of any unique forms in rat area 17. An analysis of the dendritic projections of individual non-pyramidal neurons through particular cortical laminae made possible an evaluation of common sources of dendrites present in the neuropil of each layer. Non-pyramidal cell axons were impregnated only in small numbers. Spinous multipolar axons invariably exhibited a descending main branch, while the axons of bipolar neurons were distributed in a narrow vertical field. Axonal patterns of remaining cell types, including Golgi type II arborizations, did not appear to correlate consistently with dendritic morphology. Axons of the basket cell type and "horsetail" axons associated with double bouquet cells of Cajal's original type were not impregnated.


Chronic renal failure in diabetes: survival with hemodialysis vs. transplantation.

Clinical observations for 49 diabetic patients who required chronic hemodialysis or renal transplantation during a four year period are presented. Twenty-seven dialysis patients had a two year cumulative survival of 74% compared to 54% for 22 transplantation patients. The cumulative survival of live-related donor recipients (77%) was similar to that of the dialysis group and significantly better than that of cadaveric allograft recipients (36%). While the incidences of cardiomegaly and of motor neuropathy were high among live-related donor recipients, dialysis patients more often demonstrated peripheral vascular disease. Causes of death in hemodialysis patients included cardipulmonary arrest and patient decision to discontinue therapy; in the transplantation group included cardiopulmonary arrest, sepsis, and stroke. Living-related transplantation remains the preferred mode of therapy because of the potential for rehabilitation. In terms of patient survival, the risks of cadaver transplantation must be weighted against the discomforts of chronic dialysis.


The effect of diazepam on the recovery of endogenously depressed patients.

Forty endogenously depressed patients given standard antidepressants and/or electroconvulsive therapy in a general hospital psychiatric ward were assessed using the short form of the Beck Depression Inventory and the Hamilton Depression Rating Scale. The patients were then randomly assigned to two groups: Under double-blind conditions, one group received in addition 20 mg diazepam per day and the other, identical placebos. No additional benzodiazepines were prescribed during the study. Standard therapy with tricyclic antidepressants or electric shock was determined by the patients' own psychiatrists. The assessments were repeated after ten days of diazepam or placebo treatment. Fourteen patients (seven in each group) also completed the Buss-Durkee Hostility Inventory. Results of the Beck Depression Inventory indicated that the addition of diazepam retarded the improvement of the patients receiving only tricyclic antidepressants but had no effect on the recovery of patients receiving electroconvulsive therapy.


Auricular and Periauricular surgery.

Various surgical procedures are described for common disorders on and about the ear. Photographs are presented to show how they are performed. Applicable surgical subtleties and precautions are discussed.


The face lift as an office procedure.

Salient techniques for doing a face lift as an office procedure are described. Heavy preoperative sedation and analgesia are recommended. The various incisions required are diagramed and the extent of undermining required is depicted. The need for absolute hemostasis is stressed. The type of pressure dressing advised is pictured. Some useful variations practiced by other skilled plastic surgeons are presented.


Clinical application of the functional matrix: mandibular reconstruction and monobloc functional therapy.

A 13-year-old black girl with an extensive ameloblastoma of the mandible required subtotal mandibular resection and immediate reconstruction with use of an autogenous, ipsilateral, corticocancellous graft from the iliac crest and monobic functional therapy. Twenty-seven months later, clinical evaluation showed normal mandibular function and no facial asymmetry. Bony architectural changes after 27 months included reformation of the coronoid process, angular process, and external oblique ridge; preservation of mandibular length and thickness; and maintenance of the alveolar ridge height. The reformation of the coronoid process suggests the importance of the functional matrix (temporalis muscle) in skeletal tissue growth. The monobloc appliance in conjunction with bone grafting is an effective means for transmission of biophysical stimuli and for maintenance of appropriate stress for remodeling of bone in the mandible.


Immunotherapy in cat-induced asthma. Double-blind trial with evaluation of bronchial responses to cat allergen and histamine.

Ten asymptomatic patients with normal pulmonary function were selected for a double-blind trial of immunotherapy in cat-induced asthma. Each patient had a positive prick test to cat pelt extract and also a positive bronchial challenge response to the same extract. Patients were randomly assigned to active treatment or placebo groups and received weekly or biweekly injections over a 3 to 4-month period. The 5 patients who received the active treatment received a cumulative dose of cat pelt extract that ranged from 16.4 to 44.8 mg of total solid containing 1.7 to 4.7 mg of cat allergen 1. Apparent systemic reactions were observed in 3 patients who received the placebo and 3 patients who received the active treatment. The 5 patients who received the active treatment showed a reduction in skin reactivity to cat pelt extract as well as a significant mean reduction in bronchial sensitivity to the same extract. The 5 patients who received the placebo showed no significant changes in skin reactivity or bronchial sensitivity to cat pelt extract. Bronchial response to histamine did not change significantly in either the active treatment of the placebo group.


Severe impairment of dietary folate utilization in the elderly.

Absorption of folates, vitamin B6, pantothenate, and riboflavin from a natural food source (yeast) and from synthetic folylmonoglutamate was studied in 24 elderly subjects (age range, 73 to 101 years) and in 12 healthy younger subjects (age range, 24 to 42 years). All subjects absorbed riboflavin, vitamin B6, and pantothenate from yeast. Ingested folypolyglutamates (the preponderant folates in yeast) proved to be a very poor source of folates for the elderly subjects, whereas synthetic folylmonoglutamate served as a good source. In the younger subjects, yeast was a significant folate source. It is suggested that the folate deficits so common in the elderly are caused by impaired ability to obtain folate from ingested foods. The resultant folate deficit may induce changes in the epithelial structure and enzyme secretion of the small bowel, which may further exacerbate folate malabsorption.


Lactulose syrup assessed in a double-blind study of elderly constipated patients.

The laxative effects of 50% lactulose syrup and 50% glucose syrup were compared in a 12-week, double-blind study of 47 elderly constipated patients living in a nursing home. The dosage was 30 ml daily taken at bedtime; it was reduced to 15 ml if the initial dosage produced two or more bowel movements daily. The number of bowel movements during treatment in comparison to pretreatment was significantly increased in the 42 patients (19 lactulose, 23 glucose) who completed at least 8 weeks of the study. Laculose was superior to glucose in the mean number of bowel movements per day (p less than 0.02) and in the percentage of days in which at least one bowel movement occurred (p less than 0.05). Reduction in the severity of each 5 symptoms (cramping, griping, flatulence, tenesmus, bloating) was greater with lactulose. For relief of all 5 symptoms, lactulose was significantly more effective than glucose (p less than 0.04). The striking reduction in the number of fecal impactions (only 6 in the lactulose patients vs 66 in the controls) was highly significant (p less than 0.015). The lactulose patients needed fewer enemas than did the controls. No abnormal values were observed in laboratory tests.


Bone grafts: a review of the literature.

Dr. Glidear discusses bone transplantation and the incorporation of transplanted bone into the host. The graft itself plays a role in its incorporation into the host by either attracting invading vessels from the bed or by forming anastomoses between itself and the host vascular channels. Vessels can invade a bony graft only when shear is eliminated or greatly minimized. The most frequent cause of transplantation failure is infection.


Community mental health centers: politics and therapy.

This paper traces the development of theory and public awareness of mental health from 1900 to 1960, with particular stress on the rise of social psychiatric models and the impact of events in and around World War Two. The federal legislative history of the Community Mental Health Center (CMHC) program through 1976 is then outlined with regard to particular social problems (e.g., alcoholism) and to domestic politics as they influenced the program's regulations and mandates. A brief critique of the CMHC program from both viewpoints follows, with emphasis on poor administration, lack of community control, and poor evaluation and accountability. This is the basis of an argument for a more egalitarian, explicitly political viewpoint and methodology as a start toward solving problems that chronically afflict the mental health system.


Fine structure of noradrenaline containing nerve fibers in the median eminence of female rat demonstrated by in situ fixation of potassium permanganate.

Fine structure of noradrenaline (NA) terminals in the internal layer of the median eminence (ME) was elucidated by in situ fixation of potassium permanganate. The NA terminals in ME contained relatively large cored vesicles (about 1000 A) together with a large number of small core vesicle (400-600 A). The most dominant feature that NA terminals show is the closed contact with ependymal cells and their processes. In the bulging abutting on the contact membrane, some of core vesicles fused or touched the membrane appearing to release the content into intercellular space or take up NA from the space. This finding shows that the NA terminals abutting on the ependymal cells and their processes have a close contact with them and give an important influence on regulation of ependymal function. Our present study further revealed that the NA terminals also had a close contact not only with the axon terminals in which ordinary spherical non-cored vesicles were exclusively present but also with the axonal fibers of neurosecretary system. These findings would infer complicated function of NA in ME.


Life change and illness studies: past history and future directions.

A selected review of life changes and illness studies is presented which illustrates both the diversity of samples that have been tests in these studies and the generally positive results which have been obtained. Although early (retrospective) work in this field led to simplistic explanations, later (prospective) studies have begun to document the several intervening variables which occur between subjects' recent life change experiences and their subsequent symptomatology and disease. A life change and illness model is presented to illustrate key intervening variables. The authors believe that future research in the field of life change and illness should concentrate on further delineations of these intervening variables--an effort necessitating active collaboration between social and medical scientists.


Epidemiologic research in hypertension: a critical appraisal.

Epidemiologic studies of blood pressure have been conducted for over twenty-five years, but the results of this research can only be described as modest. The basic epidemiologic and demographic description of blood pressure distributions in human populations remains problematic and psychosocial studies have not yielded clear and solid hypotheses pointing the way to future research. There is no doubt that blood pressures vary among and between population groups, and there seems little doubt that variations in life-style are associated with these differences. It is puzzling that we have failed to discern systematic and patterned relationships among these variables. It is suggested that a more useful approach to research in this field would be to distinguish factors that affect general susceptibility to becoming ill from those that initiate or maintain particular disease states.


Migration of radioactive products through alimentary chains (a review).

Data on the mobility of radioactive products in biocoenoses are presented and the main routes of their migration are discussed. Data on radioactive contamination of food products of vegetable, animal and marine origin are mentioned. It has been demonstrated that technological processing of raw material and culinary preparation of food products results in a considerable reduction in their contamination. Data on the amounts of radioactive nuclides taken up by the human organism in consequence of local and global fallout of radioactive products are reported.


Incidence of Q fever in two inadequately investigated occupational groups.

In 1973 the authors investigated the incidence of Q fever serologically by means of the reaction of complement fixation (RCF) and the method of immunofluorescent titration (MIFT) in two inadequately investigated occupational groups--communal workers from the town of Russe and medical workers in obstetric departments of several towns in North Bulgaria. In addition, they carried out comparative studies in order to characterize the incidence and the degree of affection from the same disease in other persons exposed and not exposed at work in the same area--transport workers and blood donors. Out of 198 communal workers, 91 (45.95 +/- 3.54%) had positive titres for Q fever (1:8--1:512). A high incidence of Q fever was established in dustmen (61.40%), sweepers (46.55%) and drivers of dust cars (38.00%), i.e. persons collecting and rendering harmless the garbage of big town. Out of 174 medical workers in obstetric departments 65 (37.36% +/- 3.78%) were positive in titres 1:8--1:512. A high incidence of Q fever was established in obstetricians (57.14%), midwives (38.11%) and hospital attendants (34.38%), i.e. persons providing medical care for pregnant women or women in childbirth. In both groups the occupational hazard increases with the length of service. Out of 244 transport workers 82 (33.60% +/- 3.02%) were positive for Q fever, and out of 237 blood donors 19 (8.01 +/- 2.54%) were serologically positive for Q fever. The authors suggest continued investigation of these two occupational groups.


B lymphocyte precursors in human bone marrow: an analysis of normal individuals and patients with antibody-deficiency states.

Lymphoid cells containing cytoplasmic IgM but lacking stable surface IgM are believed to be the direct precursors of B lymphocytes. We have characterized these pre-B cells in the bone marrow of normal individuals and patients with a variety of immunoglobulin deficiencies or hematologic disorders by using immunofluorescence and autoradiography. Pre-B cells comprised 5.8 +/- 5.7% of lymphoid cells in normal bone marrow. Eleven patients with infantile X-linked agammaglobulinemia (X-LA) lacked B lymphocytes but had a normal frequency (3.8 +/- 3.6%) of bone marrow pre-B cells. A smaller proportion of marrow pre-B cells from patients with X-LA were engaged in spontaneous DNA synthesis than was found for normal controls. In individuals other than the group with X-LA, the number of circulating B cells was positively correlated with the frequency of marrow pre-B cells. These results indicate that patients with X-LA have a defect in maturation of pre-B cells, and suggest that some patients with acquired B lymphocyte deficiency may have lost the capacity to generate pre-B cells from stem cells.


A two-step centrifugation procedure for the purification of sheep erythrocyte antigen-binding cells.

A two-step centrifugation procedure has been developed to isolate greater quantities of highly purified sheep erythrocyte antigen-binding cells (ABC) than previously possible. The first step involves partially separating sheep erythrocyte rosettes from unrosetted lymphocytes by their difference in buoyant density on Ficoll-Hypaque. Subsequent passage through a linear 5 to 10% Ficoll gradient produces further purification of rosettes by sedimentation velocity. Approximately 4.5 X 10(6) ABC can be obtained at 50 to 100% purity from 10(9) immune spleen cells (5 days post-immunization) and 1 X 10(5) ABC at 20 to 40% purity from 10(9) nonimmune spleen cells. The purified ABC from 5-day immune animals are 80 to 90% B cells and 10 to 20% T cells, and represent between 30 and 40% of the original ABC in the spleen cell population. Less than 0.2% of the purified ABC are plaque-forming cells (PFC) and less than 2% have intracellular immunoglobulin (Ig) or J chain. The quantities of ABC obtained are sufficient for investigating biochemical parameters of antigen-induced lymphocyte activation and for direct analysis of the surface isotypes found on antigen-binding cells after immunization.


Impairment of cell-mediated immunity in mutation diabetic mice (db/db).

Mutation diabetes in the mouse occurs in the C57BL/Ks strain. All homozygous animals (db/db) develop obesity, elevated blood sugar levels and increased or normal blood insulin concentration. The defects in cellular immunity in db/db mice and their littermate controls were examined both in vivo and in vitro. Significant suppression of delayed footpad swelling and first and second set skin allograft rejection time were observed. In addition, DNA synthesis in spleen cells after nonspecific mitogen stimulation was markedly inhibited. Diabetic animals with a mean blood sugar of 512 +/- 101 mg/100 ml did not respond to exogenous insulin therapy by lowering their blood sugar levels or reversing the defect in DNA synthesis. Adding insulin to spleen cell culture in vitro had no demonstrable effect on their response to mitogens. Thus, mutation diabetic mice with their known defect in the peripheral utilization of insulin have markedly suppressed cell-mediated immune mechanisms.


Senile cardiac amyloid: demonstration of a unique fibril protein in tissue sections.

Antisera were raised against degrading amyloid fibrils isolated from the heart of a patient with senile cardiac amyloidosis (SCA), and from a medullary carcinoma of the thyroid (MCT). The antisera were absorbed and used in indirect immunofluorescence to identify an amyloid fibril protein (ASCA) in heart tissue from patients with senile cardiac amyloidosis and to identify the amyloid fibril protein (AMCT) found in association with medullary carcinomas of the thyroid. Absorbed anti-ASCA antiserum did not react with normal tissue such as heart, liver, spleen, and striated muscle, or with amyloid tissue known to contain amyloid fibril proteins AA, AlambdaI, AlambdaIV, AlambdaV, AMCT or with pancreatic tissue containing islet amyloid deposits. The reactions with senile amyloid he,rt tissue could be blocked completely by degraded amyloid fibrils extracted from senile amyloid heart tissue or by amyloid fibril protein ASCA isolated from such fibrils. The anti-AMCT antiserum showed a similar specific reaction restricted to amyloid associated with MCT. In addition, antisera specific for amyloid fibril proteins AA, AlambdaI, AlambdaIV, and AlambdaV failed to react with senile cardiac amyloid, pancreatic islet amyloid, or medullary thyroid amyloid.


Influence of the major histocompatibility complex in the pig (SLA) on serum haemolytic complement levels.

Sera from fourty-six non-congenic adult pigs from one inter-related herd were tested for their haemolytic serum complement levels. A comparison of these data with the major histocompatibility complex (SLA) genotypes of these animals resulted in the demonstration of highly significant differences in lytic activity between sera obtained from animals of two distinct SLA haplotypes. These data showed a high correlation between the SLA complex and the level of haemolytic activity.


Localization of the collagenous component in skin basement membrane.

Antibodies to type IV collagen were produced by immunizing rabbits with a basement membrane collagen obtained from a transplantable mouse tumor. Using specifically purified antibodies, type IV collagen was localized ultrastructurally to the basal lamina part of the basement membrane zone.


Characterization of pemphigus antigen.

New factors controlling the reaction of pemphigus antibody and the pemphigus antigens were defined by studying the action of antibody on section prepared from human skin. The pemphigus antigenicity was protected by adding dithiothreitol and CaCl2 in buffer. Blocking studies of the -SH groups by sulfhydryl reagents rapidly decreased the reactivity of antigens. In addition pretreatment calcium chelating agents also decreased this reactivity.


Use of the Y-body for identification of skin source on a successfully grafted burn patient.

The use of the interphase male Y-body (fluorescent Y chromosome segment) technique with cryostat sections of both fresh and frozen-stored skin biopsies is described. A female burn patient appeared to retain her donor homografts, thereby negating the need for autografts. Since a retained homograft of this sort challenged our understanding of immunologic barriers, we applied the Y-body technique to cutaneous biopsies obtained from the patient's burn area that had been homografted with skin from a male donor, as well as control biopsies from the patient's unburned skin and normal control male and female skin. Based on clinical and cytogenetic observations, it was concluded that the most reasonable explanation for this case was that the regenerating tissue at the graft site was that of the recipient and not that of the originally grafted male skin.


An immunofluorescence assay for double-stranded DNA antibodies using the Crithidia luciliae kinetoplast as a double-stranded DNA substrate.

Circulating antibodies to dsDNA are found predominately in SLE and are seen most often in those patients with active systemic disease, particularly severe lupus glomerulonephritis. An IIF technique for measuring these antibodies has recently been described. This uncomplicated assay employs the kinetoplast of the nonpathogenic hemoflagellate Crithidia lucilliae as a dsDNA substrate. We herein report our experience with this assay, emphasizing the methodology and interpretation of results. Although slightly less sensitive than a radioimmunoassay, we find that this IF technique is a specific and reliable qualitative method for detecting anti-dsDNA. An estimate of the amount of DNA antibody present can be obtained by serum titration. The test was positive in only two patient groups tested, SLE (48%) and MCTD (20%).


A rapid and quantitative assay of phagocytosis-connected oxygen consumption by leukocytes in whole blood.

We have devised a simple method for quantitative assay of the phagocytosis-connected oxygen consumption by human peripheral leukocytes. Time-consuming and leukocyte-debilitating cell fractionation procedure is avoided by the use of whole blood as assay material. Heparinized venous blood was treated with CO and put into a cuvette. A Clark-type oxygen electrode was equipped to the cuvette. A decrease in oxygen concentration in the whole blood was induced by the addition of bacteria. The decrease was exclusively dependent on phagocytizing leukocytes. All procedure can be performed within half an hour. Blood from boys with CGD showed little or negligible increase in oxygen consumption, and that from their mothers showed about 30% activity of control blood.