Comparison of three procedures for biochemical testing of anaerobic bacteria. The Analytab Products, Inc. (API), anaerobic multitest microsystem (MICRO) was compared with the Center for Disease Control conventional (CONV) thioglycolate (supplemented with hemin and vitamin K1) system and with pre-reduced anaerobically sterilized (PRAS) media as recommended by the Virginia Polytechnic Institute. Growth from a solid medium was suspended to produce standard inocula. Substrates included 16 carbohydrates, indole, urea, gelatin, and esculin. API strips were inoculated in air and incubated in GasPak (BBL) jars. MICRO tests were read at 1 and 2 days. CONV tests at 1, 2, and 7 days, and PRAS tests at 3 weeks. One hundred thirty well-characterized strains of anaerobes (76 gram-negative rods, 16 cocci, 26 gram-positive nonsporeforming rods, and 12 clostridia), including 48 reference strains, were studied. Of 2,600 tests performed, 2,085 (80.2%) showed agreement with all three methods. There was 90.9% agreement between the MICRO and CONV, 84.9% between the MICRO and PRAS, and 84.6% between the CONV and PRAS tests. All MICRO tests were reliable except for indole, which was not sensitive enough, and gelatin, which was very insensitive. The MICRO system permits performance of biochemical tests at the workbench in the average clinical laboratory without the need for expensive equipment and time-consuming procedures.


Practical aerobic membrane filtration blood culture technique: clinical blood culture trial. During the course of preliminary clinical trials of an improved membrane filter blood culture system, filter plugging produced by a gelling of the lysing solution was observed when the patients had high leukocyte counts. A solution of streptokinase-streptodornase (Varidase, Lederle Laboratories) dissolved or prevented the gel and permitted rapid filtration without plugging. With streptokinase-streptodornase incorporated in the filtration procedure, a comparison of several culture systems was carried out on 176 blood cultures. Brucella broth with and without sodium polyanethol sulfonate, a prereduced osmotically stabilized broth, pour plates, and an improved aerobic membrane filter system were compared. The membrane filter system yielded 29 of the total of 37 positive cultures, far surpassing all other systems. Eight of these cultures were detected first by the filter technique, and 13 were positive only in this system. Nineteen of the 37 positive cultures were from patients on antimicrobial agents. Fourteen of these were detected by the filter, twice the number detected by any of the other systems.


Presumptive speciation of Streptococcus bovis and other group D streptococci from human sources by using arginine and pyruvate tests. A simplified method for speciation of group D streptococci is described. A total of 4,156 streptococcal isolates from human clinical material was tested for ability to hydrolyze esculin in the presence of 40% bile, ferment pyruvate, hydrolyze arginine, and grow in media containing 40% bile or 6.5% NaCl. Streptococci which hydrolyzed esculin in 40% bile, but which did not hydrolyze arginine, were also tested for their ability to ferment raffinose or sorbose. Sixty percent (2,503) of the isolates hydrolyzed esculin in the presence of 40% bile and were thus presumptively identified as group D. By application of the other criteria, 84% of these were speciated as Streptococcus faecalis, 7% were speciated as S. faecium, 6% were speciated as S. bovis, 2% were speciated as S. avium, and 1% were not identified. This scheme was shown to be both reliable and practical for use in the diagnostic laboratory. S. avium and S. bovis isolates were characterized, and 18 S. bovis isolates from patients with bacterial endocarditis were compared physiologically with 151 isolates of this species from other sources.


Recognition of Pseudomonas pickettii in the clinical laboratory: biochemical characterization of 62 strains. Pseudomonas pickettii strains were studied to determine the characteristics essential for their identification in the clinical microbiology laboratory. Preliminary investigations indicated that these glucose-oxidizing, denitrifying, gram-negative rods were quite similar to an unclassified group of clinical isolates designated VA-2. Gas liquid chromatography of trimethylsilyl derivatives of whole cell hydrolysates of P. pickettii and VA-2 strains yielded nearly identical elution profiles. The VA-2 cultures were concluded to be probable strains of P. pickettii. A method is presented for differentiating cultures of P. pickettii from other similar bacteria encountered in clinical specimens.


Evaluation of blood culture media supplemented with sucrose or with cysteine. A total of 5,883 blood samples from patients with suspected bacteremia were inoculated concurrently into each of three media under vacuum with CO2: tryptic soy broth (TSB) with sodium polyanetholesulfonate (SPS), TSB with SPS and cysteine, and TSB with SPS and sucrose. There were 395 positive cultures, excluding presumed contaminants. No significant differences were noted with the addition of cysteine to TSB with SPS, and no streptococcal mutants requiring thiol groups were isolated. Haemophilus, Staphylococcus aureus, and bacteriodaceae were isolated more frequently (P less than 0.05) in the absence of sucrose. The addition of sucrose to TSB containing SPS did not significantly increase the rate of positivity or the time interval to detection of positivity of any group of bacteria.


Purification of Japanese encephalitis virus vaccine by zonal centrifugation. A large-scale procedure for purification and concentration of Japanese encephalitis virus vaccine by a continuous-flow isopycnic banding technique in a sucrose density gradient solution, using a K-III zonal centrifuge rotor, is presented. The quality of zonal-purified vaccine was compared with commercial and Japanese National Institutes of Health reference vaccines for antigenicity, immunodiffusion, and allergic encephalitogenicity tests to show its high purity.


Rapid method to detect rubella immunoglobulin M and immunoglobulin A antibodies. Immunoglobulin (Ig) G was removed from serum specimens by precipitation with gamma chain-specific anti-human IgG of rabbit origin. The remaining rubella virus-specific IgM (and IgA) antibodies were then detected by the rubella hemagglutination-inhibition test. This procedure has proven to be as reliable as estimations carried out with IgM fractions separated on a sucrose density gradient.


Biotypes of Proteus rettgeri.

Biotypes of Proteus rettgeri. Examination of 729 isolates of Proteus rettgeri showed that 674 reacted positively in tests for phenylalanine deamination, indole production, growth on citrate, and acid production from meso-inositol, and negatively for L-ornithine decarboxylation and acid production from lactose, maltose, D-xylose, and L-arabinose. Only 51 isolates differed in one, and four differed in two of these ten reactions, which were taken as the core characteristics of the species. On the basis of additional tests (acid production from salicin, L-rhamnose, D-mannitol, adonitol, and D-arabitol), the 729 isolates could be separated into five groups. Goups 1, 2, 3, and 4 could be further separated on the basis of the reaction with meso-erythritol, and group 5 could be subdivided on the basis of reaction with D-mannitol. Two metabolically distinct kinds of P. rettgeri were recognized. Isolates of the first kind (groups 1, 2, 3, and 4) each utilized both adonitol and D-arabitol and most utilized meso-erythritol. Isolates of the other kind (group 5) were negative with the three polyhydric alcohols but resembled, in their reactions, some strains of Providencia stuartii. These may be intermediates between P. rettgeri that catabolize these substrates and the Providencia.


Bacteriology of normal and infected conjunctiva. The conjunctivas of 273 inflamed eyes were cultured by both aerobic and anaerobic techniques. Isolations were obtained from 267 (97.8%) of the eyes. Aerobic organisms were isolated from 237 (86.8%) of the conjunctivas. Staphylococcus aureus from 63 (23.1%) of the diseased eyes was the aerobic pathogen most often isolated. Anaerobic bacteria were isolated from 172 (63.0%) of the conjunctivas. Propionibacterium acnes was isolated from 126 (46.2%) of the conjunctivas, and Peptostreptococcus species were isolated from 80 (29.3%) of the conjunctivas. Comparison with the bacteriology of 96 normal eyes showed that anaerobes play a much greater role as etiological agents of conjunctivitis than formerly believed. When anaerobic bacteria were isolated, they appeared on the average in 7 days, indicating that they may be missed by ordinary bacteriological culturing.


Quantitative assay of dermatophyte-infected guinea pig skin scales. A technique to obtain homogeneous suspensions of fungal particles in scrapings from infected skin was devised that permits delivering uniform inocula to culture media being evaluated for suitability as primary isolation media for dermatophytes. The technique consisted of trypsinization of the scales, utilizing scrapings from lesions experimentally induced with an isolate of Trichophyton mentagrophytes var. granulare on guinea pigs. Optimal conditions for dislodging the fungal particles were assessed by colony counts. One-hour treatments with 0.5% (1:300) trypsin yielded the highest counts.


Effect of aerobic and anaerobic atmospheres on isolation of organisms from blood cultures. Blood was cultured in two vacuum bottles containing Columbia broth. Filtered air was admitted to one bottle (aerobic); the unvented bottle was considered anaerobic. Cultures were incubated at 35 C until growth occurred or for at least 7 days. Of 744 organisms isolated, 50% were isolated from both bottles, 30% from the aerobic bottle only, and 20% from the anaerobic bottle only. These results indicate the need for use of both an aerobic and anaerobic bottle for blood cultures.


Oxygen tolerance of fresh clinical anaerobic bacteria. The oxygen tolerance and sensitivity of 57 freshly isolated anaerobic bacteria from clinical specimens was studied. All the organisms tolerated 8 h or more of exposure to oxygen in room air. Growth of the isolates in increasing oxygen concentrations demonstrated that the 57 isolates varied in oxygen sensitivity from strict to aerotolerant anaerobes. Comparison of the oxygen tolerance and sensitivity showed that the most tolerant organisms (best survival after prolonged exposure) included anaerobes capable of growth at only 0.4% or less O2 (strict) as well as those able to grow in as much as 10% O2. The least tolerant were predominately strict anaerobes. Decrease in the inoculum size from a concentration of 10(8) to 10(6) colony-forming units per ml had only a minor effect. The data indicate that the brief oxygen exposure with bench techniques in clinical laboratories would not be deleterious to the anaerobic bacteria present in clinical specimens.


Significant enhancement of T-strain mycoplasmal growth on supplemented Shepard's A6 differential agar. Growth of T-strain mycoplasmas was significantly improved by supplementing Shepard's A6 medium with sodium thioglycolate, cysteine hydrochloride, mercaptoethanol, or dithiothreitol.


Virulence factors of biotypes of Staphylococcus epidermidis from clinical sources. The biotyping scheme of Baird-Parker was applied to cultures of Staphylococcus epidermidis from patients. In all, 63.6% of 228 cultures belonged to biotype 1, followed by biotypes 4, 3, and 2 in decreasing order of incidence. When classified according to clinical source of isolation, cultures of S. epidermidis were most frequently isolated from urine, with 39.5% of 228 cultures from this source. Each of the four biotypes was distributed throughout all nine catagories of clinical sources. The production of virulence factors was based on the results of three groups of tests: (i) deoxyribonuclease, urease, gelatinase, caseinase, and lysozyme production; (ii) lipolytic activity on the tweens; and (iii) hemolysin production. Enzymatic activity was highest for organisms in biotypes 1, followed by biotypes 3, 4, and 2 in decreasing order. Of the 228 cultures, 76.3% were lysed by lysostaphin. Resistance to antibiotics was highest for tetracycline, ampicillin, and penicillin, with rates of 54.8, 69.3, and 81.6%, respectively. The role of S. epidermidis as an etiological agent was studied by analyzing the laboratory and clinical data of 80 patients selected at random with bacteriuric S. epidermidis. Organisms in biotype 1 were most commonly associated with urinary tract infection. The significance of certain biotypes of S. epidermidis as opportunistic pathogens among compromised hosts in a hospital environment is discussed.


Identification of streptococci: use of lysozyme and Streptomyces albus filtrate in the preparation of extracts for Lancefield grouping. A combination of lysozyme and Streptomyces albus filtrate has been shown effective in extracting group-specific antigen for all commonly occurring serologically groupable streptococci. A prospective comparison of this method with that of Rantz and Randall (1955) for grouping 761 clinical isolates has confirmed its accuracy, which in our hands exceeded that of the latter more complicated method of serogrouping. Its rapidity and simplicity and the relatively low cost of the reagents involved make it practical for routine use in clinical bacteriology laboratories.


Passive transfer of immunity into leprosy patients by transfusion of lymphocytes and by transfusion of Lawrence's transfer factor. About 1,200 million viable lymphocytes from normal but lepromin- and tuberculin-positive human beings were transfused in four patients of lepromatous and one of tuberculoid leprosy three times at monthly intervals. Three patients of lepromatous leprosy suffered from erythema modosum, whereas the other two developed severe reaction whenever put on the smallest dose of dapsone. In one patient of lepromatous leprosy, minimal improvement or none was observed, whereas in the remaining three cases of lepromatous and one of tuberculoid leprosy, clinical, bacteriological, as well as histological improvement occurred. Two of the five patients started to tolerate the dapsone during the period of study. The present study indicates that immunotherapy might have a definite role in the management of the disease especially in cases with erythema nodosum. Lawrence factor, prepared from leucocytes of normal donors, was transfused three times into four lepromatous leprosy patients who were intolerant to anti-leprosy drugs. The donors were healthy but were tuberculin and lepromin (Mitsuda) positive. The clinical, histological, bacteriological (morphological index), and immunological assessments of the patients were performed before and 5 months after starting the immunotherapy. In two patients conversion of Mitsuda reaction occurred, but there was no appreciable improvement in the clinical, histologic, and bacteriologic status of these patients.


Detection of fungi in blood cultures.

Detection of fungi in blood cultures. In a retrospective study covering the period January 1972 to June 1974, recovery rates of bacteria and of fungi were generally equivalent with tryptic soy broth, Thiol, thioglycolate, and Columbia broth media (all under vacuum with carbon dioxide and sodium polyanetholesulfonate). An additional biphasic medium consisting of brain heart infusion broth and a brain heart infusion agar slant, which was inoculated only where fungal sepsis was suspected clinically, yielded significantly higher recovery rates of fungi. There were 29 instances of cultures with fungi in both the biphasic and broth media, 80 instances of cultures with fungi only in the biphasic medium, and no instances of fungi only in the broth media. The isolates were as follows: Candida albicans, 74; C. parapsilosis, 20; C. tropicalis, 16; Torulopsis glabrata, 18; Torulopsis sp., 1; Cryptococcus neoformans, 12; C. laurentii, 2; and Histoplasma capsulatum, 16. Despite routine subcultures of the broth media to chocolate blood agar within 24 h of inoculation and after 5 days of incubation, detection of fungemia was significantly improved by the use of a biphasic medium.


Cultural and biochemical characteristics and fatty acid composition of Pseudomonas diminuta and Pseudomonas vesiculare. The cultural characteristics, biochemical activity, and cellular fatty acid composition of Pseudomonas diminuta and Pseudomonas vesiculare provided means for differentiation of these closely related species. Broth cultures of P. diminuta showed turbid growth and a distinct surface pellicle after 24 h at 35 C. P. vesiculare had no pellicle, and only light, diffuse growth was observed. All strains of P. vesiculare oxidized maltose and hydrolyzed esculin to varying degrees; P. diminuta was negative in these tests. The cellular fatty acids of these two species were similar, except that P. diminuta possessed a C19 cyclopropane acid which was not detected in P. vesiculare.


Characteristics of Pasteurella multocida of human origin. Physiological, serological, morphological, and cultural differences were observed among 30 Pasteurella multocida cultures of human origin. The usual variations in the fermentation of glycerol, lactose, sorbitol, trehalose, and xylose were observed. Unlike most P. multocida, two cultures did not produce indol. Six serotypes were found. In addition to the widely recognized iridescent, blue, and watery mucoid (circular) colonies, punctiform colonies were observed. None of the cultures were pathogenic for turkeys. Results of the study indicate that one should be aware of the many variable characteristicx of P. multocida of human origin to facilitate indentification.


Characteristics of yellow-pigmented nonfermentative bacilli (groups VE-1 and VE-2) encountered in clinical bacteriology. The morphological and physiological characteristics of 20 strains of motile, gram-negative, yellow-pigmented  oxidative bacilli (groups VE-1 and VE-2) isolated in clinical bacteriology are described. Electron micrographs demonstrate the polar multitrichous flagella of group VE-1 and polar monotrichous flagella of group VE-2. Data obtained from guanine plus cytosine ratio studies of 56.8% for VE-1 and 68.9% for VE-2 distinguish the two groups of bacteria.


Viral studies on human brain culture retrieved from cold storage. Tissue cultures established from brain tissues of six patients with various chronic, degenerative diseases did not show evidence of viral agents. Sucrose gradient analysis of radioactively labeled culture fluid from two patients failed to reveal the presence of any incomplete or defective virus. Extracellular fluids from three brain cultures contained a factor able to stimulate deoxyribonucleic acid synthesis of Vero cells.


Cellular fatty acids of Alcaligenes and Pseudomonas species isolated from clinical specimens. The cellular fatty acid composition of 25 clinical isolates of Alcaligenes and Pseudomonas was determined by gas-liquid chromatography (GLC). The GLC fatty acid profiles of three species of Pseudomonas were markedly different from those of Alcaligenes. The most significant differences were the presence and relative amounts of hydroxy, branched-chain, and cyclopropane fatty acids. One of the major fatty acids in A. faecalis was a 17-carbon cyclopropane (17 delta) acid, whereas a 15-carbon branched-chain acid (13-methyl tetradecanoate) characterized isolates of P. putrefaciens. The determination of these fatty acids by GLC provides a rapid and specific means of distinguishing clinical isolates of Pseudomonas and Alcaligenes.


Association of Pseudomonas cepacia with chronic granulomatous disease. Pseudomonas cepacia was recovered from a number of infected sites in three patients with chronic granulomatous disease of childhood. The organisms were identified on the basis of their oxidative utilization of a variety of carbohydrates and their positive beta-galactosidase and oxidase activities. They were resistant to most antimicrobial agents and moderately susceptible to chloramphenicol. Peripheral blood leukocytes isolated from two siblings with chronic granulomatous disease, including one of the patients in this series, failed to kill P. cepacia in vitro. Prolonged prophylactic and antimicrobial therapy may well have played a significant role in the colonization and infection of these patients with P. cepacia.


Use of bile-esculin agar for rapid differentiation of Enterobacteriaceae. Bile-esculin agar has been used for several years for the presumptive identification of group D streptococci. All members of the Enterobacteriaceae family will also grow on this medium, but only certain ones can hydrolyze esculin to 6,7-dihydroxycoumarin, which reacts with iron to produce a characteristic blackening of the medium. One thousand and six cultures from clinical specimens representing 20 genera were isolated and identified. Heavy inocula from fresh pure culture isolates on heart infusion agar were placed on bile-esculin agar slants and incubated at 35 C. The slants were examined at 4 h and again at 18 h for esculin hydrolysis. Shigella, Salmonella, Arizona, Proteus mirabilis, Proteus morganii, Providencia alcalifaciens, and Providencia stuartii all produced negative results. Klebsiella pneumoniae, Enterobacter aerogenes, Serratia marcescens, and Serratia rubidaea produced a positive reaction in 4 h. The other remaining eight genera exhibited varying results. The use of this medium in conjunction with triple sugar iron-lysine iron agar has been of great value in differentiating the Klebsiella-Enterobacter-Serratia group from other Enterobacteriaceae.


Photoreactivation of Pseudomonas cepacia after ultraviolet exposure: a potential source of contamination in ultraviolet-treated waters. Cells of a naturally occurring strain of Pseudomonas cepacia grown in distilled water were exposed to ultraviolet radiation. Irradiated samples incubated on membrane filters or in fluid recovery media in the absence of light showed no evidence of dark repair mechanisms. When samples were exposed to fluorescent light ranging from 50 to 950 foot candles (538 to 10,222 lux) of illumination, apparent photo-induced repair of ultraviolet injury resulted in 1- to 4-log increases in viable cells recovered.


Use of urease test disks in the identification of mycobacteria. Use of commercially available urea differentiation disks is a rapid and convenient means of determining the urease activity of mycobacteria. In the study performed, paper disks containing Ewing's Urea R Broth were compared with two other methods of testing for urease production. Over 1,500 tests were performed with recent patient isolates from which at least 15 different species of mycobacteria were identified. The tests were read at intervals for 72 h, and a comparison of test results as to rapidity and accuracy for each species was tabulated. The urea disks showed a faster reaction time when compared with the other two methods. They also showed a high percentage (79 to 100%) of reliability for each species on the first run, based on the expected response.


Micromethod for serotyping strains of Mycobacterium avium. A rapid and economical micromethod for serotyping strains of Mycobacterium avium is described. Eighty-four of 91 strains representing 20 serotypes were successfully identified.


Evaluation of tyrosine medium for the identification of Enterobacteriaceae. Use of the tyrosine medium in the identification of members of the family Enterobacteriaceae form the basis of this paper. Six hundred and twenty-three strains were tested for their property of tyrosine degradation. Three hundred and seventy-two strains were positive in 24 h and an additional eight strains were positive in 48 to 72 h. These positive strains consisted of all 344 strains of the Proteus-Providencia group, all 25 strains of Citrobacter diversus, and three strains of atypical C. freundii. Our findings suggest that tyrosine degradation is a useful property for separating out the Proteus-Providencia group and in specific identification of C. diversus.


Naturally occurring histoplasmosis in the chinchilla (Chinchilla laniger). Histoplasmosis was diagnosed histopathologically in a female chinchilla. This animal had originated from a commercial chinchilla ranch in central Missouri. Seventeen of 130 animals in the colony had died within a month's period with a respiratory illness. This animal had a history of fur chewing, but this was not true of all the other animals that had died. Histoplasma capsulatum was cultured from timothy hay used for food.


Hemolysis-in-gel test for the demonstration of antibodies to rubella virus. A rapid and simple method for the determination of rubella immunity is described. The method, which employs passive hemolysis in agarose gel, is sensitive and reproducible and does not require prior absorption of test sera to remove inhibitors. Immunoglobulin G, but not immunoglobulin M, antibodies were regularly found to give demonstrable reactions in the test. It is concluded that the hemolysis-in-gel test may provide a valuable tool, in particular for mass screening for rubella immunity.


Evaluation of the R/B and Minitek systems for identification of Enterobacteriaceae. The R/B and Minitek systems for the identification of Enterobacteriaceae were evaluated, and the results were compared with those obtained by conventional methods. Both systems were rapid and allowed correct identification of about 85% of the 294 isolates (23 species) examined. The individual biochemical reactions showed an overall agreement of 92.6% and 93.1% for the R/B and Minitek systems, respectively.


Prosthetic valve endocarditis due to Mycobacterium chelonei. Postoperative prosthetic valve endocarditis due to Mycobacterium chelonei occurred in a man after replacement of the aortic valve. The organism was isolated from blood cultures and from vegetations taken postmortem from the edge of the inserted valve. This is believed to be the first documented report of a mycobacterial endocarditis.


Stability of respiratory syncytial virus in a new bentonite holding medium. Stability of respiratory syncytial virus was determined in a new bentonite holding medium and compared with two other commonly used media. Eventual infectivity titers were similar with all three preparations.


Naturally occurring and experimentally induced mycoplasmal arthritis of cattle. Mycoplasma agalactiae subsp. bovis strain Iowa 1136 was isolated from synovial fluids of a clinical case of arthritis in cattle on pasture in Iowa. When given to calves and cows by intra-articular or intravenous injection, it caused severe and persistent joint infections with fever, lameness, and swelling of the affected joints, plus synovitis, tendonitis, and fibrinous-purulent synovial fluids of high protein content. Intramammary administration of the organism caused severe mastitis. Calves nursing the cows developed severe mycoplasmal arthritis.


Isolation and identification of mycoplasma agalactiae subsp. bovis from arthritic cattle in Iowa and Nebraska. Two strains of Mycoplasma were isolated from synovial fluids of arthritic feeder cattle and were identified as Mycoplasma agalactiae subsp. bovis by growth inhibition and fluorescent antibody tests. The strains (Iowa 1136 and Nebraska 2) could not be distinguished from known strains (Donetta and California 01) by immunoelectrophoresis or by agar gel precipitation.


Propagation and isolation of group A coxsackieviruses in RD cells. The RD cell line, derived from a human rhabdomyosarcoma, supported replication of a number of group A coxsackieviruses, including types A5 and A6 which heretofore have been propagable only in suckling mice. A number of the group A coxsackievirus types which replicated in RD cells had higher titers in this cell line than in other cell culture systems. In tests on a limited number of clinical specimens, RD cells were slightly less sensitive than suckling mice for isolation of group A coxsackieviruses, but they did permit the recovery of certain virus types which previously could be isolated only in suckling mice. Group B coxsackieviruses replicated poorly or not at all in RD cells.


Evaluation of the enteric analyzer for identification of Enterobacteriaceae. The reliability of the Enteric Analyzer for identification of Enterobacteriaceae was evaluated using biochemical results previously obtained for 291 organisms with the conventional, R/B, and Minitek systems. The instrument correctly identified 77.3% of the organisms using conventional system results, 74.2% using R/B results, and 60.5% using Minitek results. The low rate of identification with the conventional system occurs primarily because the instrument is not programmed to consider delayed biochemical reactions. The arbitrary use of 90% and 99% probabilities for test reactions also contributes to a lower percentage of identification. The Enteric Analyzer does not replace the judgment of experienced personnel in the identification of atypical bacteria, but it may prove helpful in speeding up final computer identification of typical microorganisms.


Laboratory studies of Venezuelan equine encephalitis virus in equines, Texas, 1971. During the summer and fall of 1971, epizootic and epidemic Venezuelan equine encephalitis was detected in Texas. Isolates of epizootic (IB) and vaccine (TC-83) strains were distinguished by virulence of the former for guinea pigs. Vaccine virus was isolated from 1 to 14 days after vaccination and neutralization tests demonstrated the appearance of antibody about a week after vaccination. Viremia titers of subtype IB in horses ranged from 2.2 to 8.3 log10 suckling mouse intracranial 50% lethal doses per ml. Of 101 equines from which Venezuelan equine encephalitis virus (IB or TC-83) strains were isolated, 87 had no neutralizing antibody against Venezuelan, eastern or western equine encephalitis viruses.


Improved auxanographic method for yeast assimilations: a comparison with other approaches. An improved pour-plate auxanographic method has been developed for determining the assimilation of 14 different carbohydrates by medically important yeasts. Reduction of a dye incorporated into the agar has been correlated with the growth and carbohydrate assimilation of the yeasts, allowing the speciation of many yeasts within 24 to 48 h. This technique has been found to compare more than favorably with existing yeast assimilation techniques, in terms of rapid identification, total cost, and technician time in preparing and inoculating the plates. The dye pour-plate auxanographic technique provides an easier-to-interpret, rapid, and reproducible method of mycological identification for small clinical laboratories.


Evaluation of an indirect hemagglutination test for the diagnosis of human leptospirosis. A presumptive hemagglutination test for the serological diagnosis of leptospirosis in humans is described. The antigen was prepared from a soluble alcohol extract of an andamana strain sorbed to human O-negative erythrocytes and preserved by pyruvic aldehyde fixation. In this study, the overall sensitivity of the hemagglutination test was 92% in contrast to 69% for the presumptive slide agglutination test. The specificity was 95% for the hemagglutination test in comparison with 83% for the slide test.


Prevalence of Mycoplasma hominis and Ureaplasma urealyticum (T strains) in urine of adolescents. Adolescent children were surveyed for colonization with Mycoplasma hominis and Ureaplasma urealyticum by culturing urine specimens. Rates were compared between three study groups: (i) 397 children attending parochial schools, (ii) 293 children attending an adolescent clinic specializing in adjustment problems, and (iii) 86 children attending a renal clinic. The recovery rate was higher among postpubertal girls attending the renal clinic (33%) and the adolescent clinic (26%) than among students attending parochial high school (males 2%, females 8%). Girls had approximately eightfold higher rates than boys of the same age. Isolation of Mycoplasmataceae was associated with certain sociological determinants, such as dating, cigarette smoking, and coming from a broken home, but also with abnormal findings (protein, leucocytes) in urine.


Evaluation of the Enteric Analyzer, an instrument to aid in the identification of Enterobacteriaceae. This study evaluated the Enteric Analyzer, an instrument designed to identify Enterobacteriaceae, with data obtained from the Modified r/b Enteric Differential System, other rapid identification systems, or conventional identification systems. It is programmed for 19 reactions with data obtained from Center for Disease Control publications. The instrument is very simple to use. Typical strains from 25 of 28 species were unequivocally identified. With the other three species, two choices were given. A switch allows the user to identify all species possibilities where a given biochemical pattern occurs in more than 1% of the strains or more than 10% of the strains. The instrument is useful both in the clinical laboratory and for teaching purposes.


Motility-indole-lysine medium for presumptive identification of enteric pathogens of Enterobacteriaceae. Detection of lysine decarboxylase activity is a useful supplement to reactions on triple sugar-iron (TSI) and urea agars in the initial examination of suspected pathogenic isolates from fecal cultures. Owing to the added value of motility and indole production in the differentiation of enteric pathogens, we prepared and evaluated a motility-indole-lysine (MIL) medium. The following 890 organisms were tested: 264 Shigella, 2 Edwardsiella, 182 Salmonella enteritidis, 235 S. typhi, 3 Arizona, 32 Yersinia enterocolitica, and 172 other members of the family Enterobacteriaceae. With few exceptions the MIL medium gave the same results as the standard motility, indole, and lysine decarboxylase (Moeller) test media. All discrepancies were with the indole reaction, which was weak in 2 of 67 strains of Escherichia coli and falsely negative in 6 of 32 strains of Y. enterocolitica. When both TSI agar and lysine-iron agar (LIA) slants are used in the evaluation isolates from fecal cultures, detection of H2S is duplicated. Both LIA and MIL medium detect lysine decarboxylase and deaminase activity equally well. Because of its ability to detect motility and indole production, the MIL medium is more useful than LIA when used with TSI agar. The combination of TSI agar, MIL medium, and urea agar enables reliable initial recognition of enteric pathogens of the Enterobacteriaceae.


Motility-indole-lysine-sulfide medium.

Motility-indole-lysine-sulfide medium. A medium designed for the detection of motility, indole, lysine decarboxylase and deaminase reactions, and H2S production was devised and evaluated. Results, using 157 strains of enteric pathogens, were in agreement with reference methods. When 300 isolates from fecal cultures were screened using this medium, Shigella was easily differentiated from Escherichia and more of the Proteus species, especially P. morganii, could be eliminated from further study.


Seroagglutination test for identification of Mycobacterium paratuberculosis. Twenty-nine of 30 strains of Mycobacterium paratuberculosis isolated from clinical specimens were serologically identified. No cross-reactions were observed with Mycobacterium avium serotypes 1, 2, 4, and 8.


The effects of early tectal lesions on development in the retinal gonglion cell layer of chick embryos. The possibility that the arrival of retinal ganglion cell axons in their efferent target field could influence the subsequent progress of cytomorphogenesis in the ganglion cells was tested by destroying all or portions of the primordial optic tectum at four or five days of incubation. The lesions were made essentially prior to the outgrowth of axons from the eye. Complete bilateral or unilateral destruction of the primordial tectal field did not detectably alter the morphogenesis of the inner retina or of the ganglion cells until 11 days of incubation. After this time, extensive loss of ganglion cells occurred within the retinas contralateral to the projection fields that were destroyed. The cytology of this degeneration is described as seen with the light and electron microscope. After smaller lesions involving the anterior or posterior half-tectum on one side, however, variable results occurred both in tectal development and in the retinal locus of ganglion cell degeneration observed in flat mount preparations. Many partial or punctate lesions, in fact, resulted in no detectable loss of cells from the ganglion cell layer. These results are discussed in regard to specification and possible regulatory capabilities of the tectum. Cell death in normal retinas, not previously reported in the chick, is also considered in relation to the experimental results.


Different targets of different neurons in nucleus gracilis of the cat. Efferent projections of neurons in the cat nucleus gracilis to the ventrobasal complex (VB) of the thalamus and the dorsal accessory portion of the inferior olive (IOd) were studied using tracing techniques that exploit neuronal orthograde and retrograde intra-axonal transport processes. These projections were studied in order to determine if the heterogeneity of the morphology, physiology and afferent input of the nucleus gracilis is paralleled by a similar heterogeneity in its efferent projections. In an orthograde study, 3H-leucine and/or 3H-proline were injected into the nucleus gracilis of different subjects in an attempt to label different proportions of large (GREATER THAN 18 MU) AND SMALL CELLS. This procedure permitted study of the efferent targets of the two cell types. The amount of labeling in VB was a constant ratio of the number of large cells in the nucleus gracilis that had incorporated the isotope. This relationship was not observed for the small cells. The amount of labeling in IOd was dependent neither on the number of large nor the number of small cells that had incorporated the isotope. In a retrograde study following extensive injections of horseradish peroxidase (HRP) into the thalamus, only large cells (greater than 18 mu) in the dorsal column nuclei were filled with HRP reaction product. These cells were located predominantly in clusters in the middle zone of the nucleus gracilis as well as rostrally. After injections including (but not confined to) the inferior olive, small cells located in the rostral and middle zones were filled with reaction procduct. A few large cells located ventrally in the middle zone of the nucleus gracilis and rostrally were also filled. Some of these ventrally located large cells may project in a collateral fashion both to the thalamus and to the inferior alive. The results of both the orthograde and retrograde studies are consistent with other evidence on the heterogeneity of the nucleus gracilis. These data strongly support the conclusion that the population of cells in the nucleus gracilis that projects to the thalamus overlaps with but is not identical to the population of cells that projects to the inferior olive.


Neuroplasticity of primary afferents in the neo-natal cat and some results of early deafferentation of the trigeminal spinal nucleus. The plasticity of cervical primary afferents has been investigated in young adult cats in which the trigeminal root, together with most of the ganglion, had been excised via a new approach to these structures during the first week after birth. Once degeneration debris had disappeared, bilateral ganglionectomies of the upper three cervical dorsal roots were done in five animals and the IXth and Xth roots transected on the side of the chronic trigeminal denervation in one instance; the degeneration pattern on the chronically denervated side was compared to that on the normal side. There was mild evidence of increased degeneration of the cervical afferents in the C1 segment and in the medulla (subnucleus interpolaris of V) on the chronically denervated side. The proliferation was more apparent in the kitten operated at three days of age and was only vestigial in animals operated at six or seven days; it had no appreciable tendency to extend into the large contiguous pool of denervated trigeminal neurons. There was no evidence of sprouting of the IXth and Xth nerve afferents. These observations indicate that primary afferents of the upper cervical roots and of the glossopharyngeal and vagus nerves have little neuroplastic potential even at early stages. The literature on neuroplasticity is controversial and a brief review of this together with suggestions regarding the reasons for some of these conflicts is presented. Some structural and functional effects of chronic trigeminal denervation are briefly described.


Olfactory relationships of the telencephalon and diencephalon in the rabbit. I. An autoradiographic study of the efferent connections of the main and accessory olfactory bulbs. The efferent connections of the main and accessory olfactory bulbs in the female albino rabbit have been studied using the autoradiographic method for tracing axonal pathways. Following unilateral injections of 3H-proline or 3H-leucine into the main olfactory bulb, radioactively labeled material transported intraaxonally by axoplasmic flow in an anterograde direction from soma to axon terminal is present ipsilaterally in the superficial half of the plexiform layer (IA) of: the entire circumference of the olfactory peduncle, the tenia tecta, the full mediolateral extent of the olfactory tubercle, the entire length of the prepyriform cortex, a transition area between the prepyriform cortex and the horizontal limb of the nucleus of the diagonal band, the nucleus of the lateral olfactory tract, the anterior cortical and posterolateral cortical amygdaloid nuclei (periamygdaloid areas 1, rostral half of 2, 5 of Rose, '31), and the ventrolateral entorhinal cortex (entorhinal areas 1, 2, 4, 5, 7 of Rose, '31). No subcortical or contralateral projection of main bulb efferents was found. After a unilateral injection of 3H-leucine into the accessory olfactory bulb, transported material could be followed caudally along the dorsal surface of the ipsilateral lateral olfactory tract. This heavily labeled projection is distinct from the unlabeled lateral olfactory tract and has been termed the accessory olfactory tract. Beginning at the level of the caudal third of the olfactory tubercle and extending caudally to the nucleus of the lateral olfactory tract is a group of small neurons intimately associated with the accessory olfactory tract. This cell group is referred to as the bed nucleus of the accessory olfactory tract. Projection sites of the accessory bulb include the bed nucleus of the accessory olfactory tract and layer IA of the medial nucleus and the posteromedial cortical nucleus of the amygdala (periamygdaloid areas 3, 4, PAM, caudal half of 2, 6 of Rose, '31). An additional accessory bulb efferent projection was found to enter the stria terminalis at the level of the medial amygdaloid nucleus and could be traced to a posterior segment of the bed nucleus of the stria terminalis. The autoradiographic findings indicate that the accessory olfactory bulb connects with portions of the amygdala that do not receive afferent input from the main olfactory bulb and provide evidence for the existence of two distinct and separate olfactory systems.


The patterns of projection of cortical areas 17, 18, and 19 onto the laminae of the dorsal lateral geniculate nucleus in the cat. The projection of cortical areas 17, 18, and 19 onto the laminar part of the dorsal lateral geniculate nucleus was investigated with degeneration methods and with the autoradiographic axon tracing method. In agreement with previous accounts, degenerating cortical axons stained by the Nauta method were restricted to laminae A, A1, C and to the interlaminar zones. In contrast, adjacent sections stained with the Fink-Heimer method showed fine dust like degeneration throughout all of the laminae of the nucleus. Comparisons of Fink-Heimer degeneration resulting from lesions of area 17 with that resulting from lesions of areas 18 and 19 further suggested that the area ) projection is heavier and more uniform than the projections from areas 18 and 19. Autoradiographic tracing of axons after intracortical injections of 3H-proline provided detailed demonstrations of the cortical projection patterns that confirmed the Fink-Heimer results. Following restricted injections of areas 17 or 18 the termination zones in the dorsal lateral geniculate nucleus consisted of columns of labeled tissue oriented perpendicular to the laminae of the nucleus. Area 17 was found to project heavily and uniformly throughout all of the laminae of the nucleus. The projection from area 18 also extended throughout all of the laminae of the nucleus, but was sparser and less uniformly distributed than that from area 17. Projections from area 18 distributed more heavily to the interlaminar zones and to lamina C than to laminae A, A1 C1, C2 or C3. A projection from area 19 to laminae C1, C2 and C3 was also demonstrated autoradiographically.


The connections and laminar organization ofthe optic tectum in a reptile (lguana iguana). The goals of this study were: (1) to describe the total pattern of projections from the optic tectum of Iguana iguana and Pseudemys scripta; and (2) to describe the contributions of particular lamina of the Iguana's optic tectum to this total pattern. Lesions were made in the optic tectum of the Iguana which damaged either all or only certain tectal laminae and, for comparison with the Iguana, lesions in the turtle's optic tectum were made which involved all laminae. The anterograde degeneration resulting from these lesions was stained with the Fink-Heimer ('67) method. The total pattern of projections from the optic tectum in the Iguana and the turtle is similar to that reported for representatives of other vertebrate classes. That is, the optic tectum gives rise to ipsilateral ascending projections to pretectal nuclei, to nucleus rotundus and to nucleus geniculatus lateralis pars ventralis of the diencephalon and, in addition, to a contralateral ascending pathway which courses via the supraoptic decussation to the contralateral diencephalon. Tectotectal connections and several descending pathways were also recognized in each species. The descending pathways include ipsilateral tectobulbar and tecto-isthmi pathways and a contralateral predorsal bundle. Lesions which damaged only certain tectal laminae in the Iguana revealed a laminar organization of the efferent projections. A lesion restricted to the superficial retinal-recipient layers, stratum griseum et album superficiale, resulted in degeneration in only nucleus isthmi pars magnocellularis and nucleus geniculatus lateralis pars ventralis. A lesion which involved both the retinal-recipient layers and stratum griseum centrale resulted in degeneration in only one additional structure, nucleus rotundus. A small lesion involving the deep periventricular layers as well as the superficial layers produced degeneration in the predorsal bundle and the ipsilateral tectobulbar tract as well as in the structures receiving input from the more superficial layers. These results are compared to the results of similar analyses of the superior colliculus in mammals.


Postnatal development of the cerebellar cortex in the rat. IV. Spatial organization of bipolar cells, parallel fibers and glial palisades. The ontogeny of the spatial organization of some components of the molecular layer was investigated in cerebella sectioned systematically in the sagittal, coronal and horizontal planes. There is no discernible organization in the distribution of cells of the proliferative zone of the external germinal layer (EGL) but from birth the differentiating bipolar cells of the subproliferative zone are aligned parallel to the surface and to the long axis of the folium. While they are still in or at the base of the EGL, the bipolar cells emit long processes, the future parallel fibers. The next step is the outgrowth of a vertical process which may reach the base of the molecular layer before the granule cell nucleus becomes translocated. The idea that the cell body truly migrates through the molecular layer is not supported by the observations. Bergmann glia cells are frequently seen in Golgi material in neonates but they are probably less numerous than in older infants and their processes are not as well aligned. It is only gradually that the EGL is perforated by flial endfeet which in older infants are occasionally organized into longitudinal rows. In mature cerebella the parallel fibers are separated by thin and relatively narrow, unstained spaces which are oriented in the longitudinal plane and can be traced from the pial surface to a zone just above the layer of Purkinje cells. It is postulated that these spaces are occupied by glial palisades formed by apposed thin vertical processes to which many Bergmann glia cells contribute. The alignment of these palisades is dependent on the orientation of parallel fibers. When the parallel fibers are reoriented by X-irradiation the glial palisades become correspondingly realigned. These observation indicate that the oriented growth of parallel fibers, which follows the polarization of bipolar cells, determines the spatial organization of the glial framework of the molecular layer. They also suggest that the glial palisades mediate functions that are not primarily developmental in nature.


The organisation of the spinotectal projection. An experimental study in the rat. The spinal grey projects upon the contralateral superior colliculus via two overlapping pathways in the lateral funiculus. One, more ventrally placed, is the classical spinotectal tract, which crosses immediately in the  spinal cord and remains crossed. The other, lying more dorsally, ascends homolaterally but is subject to delayed crossing in the brainstem (especially the intertectal) commissures to reach the contralateral colliculus. Both sets of fibers end only in the caudal half of the colliculus, predominantly in the stratum album intermedium. The projections from individual cord segments distribute in an ordered fashion as a series of transverse, overlapping bands, the cervical cord projecting most rostrally, the sacro-coccygeal most caudally. Additional fibers, which distribute with less marked topography, end in the lateral extreme of the stratum griseum profundum and stratum album profundum.


Retrograde changes in the nerve fibers of the dorsal spinocerebellar and spinocervical tracts in the cat. The potential retrograde changes that occur in the proximal segment of transected nerve fibers found in the region of the spinocervical tract (SCT) and in the region of the dorsal spinocerebellar tract (DSCT) after unilateral dorsolateral column (DLC) lesions were studied in kittens and adult cats. Nerve fibers in the region of the SCT and DSCT in the spinal segments caudal to the lesion were studied in transverse sections of Epon embedded material that had been fixed by vascular perfusion with a 4% buffered paraformaldehyde solution, post-fixed in osmium tetroxide, and stained with Azur II-methylene blue. T8 level DLC lesions in the 10-week old kitten resulted in a 14 to 25% decrease in mean fiber diameter in the SCT fibers (larger than or equal to 5 mu) of the T13 level. Retrograde fiber degeneration occurred in the small and large caliber fibers of the DSCT. A 15 to 30% decrease in mean fiber diameter occurred in the DSCT fibers (greater than or equal to 5 mu). The fiber density of the DSCT decreased by 20 to 30% and this could be correlated with a 20% loss of class C neurons (Loewy, '70) and a severe atrophy affecting the remaining 80% of the surviving class C neurons in the L3 segment. Mid-cervical DLC lesions in the 10-week old kitten resulted in a 10% decrease in mean fiber diameter in the SCT fibers (less than or equal to 5 mu), a 15% decrease in mean fiber diameter in the DSCT fibers (less than or equal to 5 mu), and no change in fiber density could be determined despite the finding that these lesions produced about a 10% loss of class C neurons. A T1 level lesion in a 20-week old kitten produced no change in the SCT fibers suggesting that the changes seen in the 10-week old kitten after midcervical lesions were probably age-dependent changes. A 20% decrease in mean fiber diameter (less than 5 mu) and a 6% decrease in fiber density was detected in the DSCT. T13-L1 level DLC lesions in the adult cats resulted in degeneration of some of the large and medium size fibers in the DSCT which was most noticeable at 20 to 25 days after surgery. DLC lesions did not cause degenerative changes in the SCT fibers 5 to 25 days after surgery.


Localization of masticatory motoneurons in the cat and rat by means of retrograde axonal transport of horseradish peroxidase. Topographical localization of monotoneurons supplying the masticatory muscles was investigated in the cat and rat, utilizing retrograde axonal transport of horseradish peroxidase. Following injection of horseradish peroxidase in each masticatory muscle, motoneurons labelled with peroxidase were seen to be aggregated into a cluster within the motor nucleus of the trigeminal nerve. Such clusters of peroxidase-motoneurons innervating each masticatory muscle were demarcated more sharply in kittens than in adult animals. The pattern of the nuclear representation of the masticatory muscles was found to be essentially the same in the cat and rat; it could be summarized as follows: The motor nucleus of the trigeminal nerve could be divided cytoarchitectonically into the dorsolateral and ventromedial divisions; the former was seen in almost the whole rostrocaudal extent of the nucleus, while the latter was localized at the levels of caudal two thirds of the nucleus. In the dorsolateral division, the temporal muscle was represented dorsally and dorsomedially, the masseter muscle ventrolaterally, and the pterygoid muscles ventromedially. In the ventromedial division, the anterior digastric muscle was represented dorsomedially, and the mylohyoid muscle ventrolaterally. It was also confirmed that the motoneurons supplying the posterior digastric muscle were localized in the accessory facial nucleus.


Autoradiographic localization of hormone-concentrating cells in the brain of an amphibian, Xenopus laevis. I. Testosterone. Autoradiographic methods were used to investigate locations of hormone concentrating cells in the CNS of Xenopus laevis. Both male and female frog brains contained cells. Four major hormone uptake sites were identified: the anterior preoptic area, the ventral infundibular nucleus, a dorsal tegmental area of the medulla and a presumptive motor nucleus of cranial nerves IX-X. The distribution of labelled cells was very similar for male and female brains. Available information on these testosterone uptake sites in anurans indicates possible roles in gonadotropin regulation and reproductive behavior.


Autoradiographic localization of hormone-concentrating cells in the brain of an amphibian, Xenopus laevis. II. Estradiol. Autoradiographic techniques for light microscopic examination of sex steroid retention were applied to the brains of male and female Xenopus laevis, and anuran amphibian, after 3H-estradiol administration. Estrogen was concentrated by cells in three telencephalic areas (the ventral striatum, the ventral-lateral septum and the amygdala), the anterior preoptic area, the ventral thalamus, the ventral infundibular nucleus, and in the torus semicircularis. The anterior preoptic area and the ventral infundibular nucleus contained the greatest number of labelled cells. The topography of estrogen-concentrating cells was the same in male and female brains. This fact and comparisons of 3H-estradiol with 3H-testosterone retention in Xenopus suggest that the sex steroid itself, and not the genetic sex of Xenopus determines the pattern of uptake by cells in the brain. The distribution of hormone-concentrating cells in Xenopus has many similarities to that found in birds and mammals. Preoptic, hypothalamic (tuberal), limbic forebrain and specific mesencephalic sites in all these forms contain labelled cells following radioactive sex steroid administration. Findings in Xenopus add to the argument for a phylogenetically stable system of hormone-concentrating nerve cells in limbic, hypothalamic and mesencephalic structures.


Non-randomness in the sequential formation of principal cones in small areas of the developing chick retina. Systematic variation in the stage of development within small retinal areas could account for the overlap between the times of formation of the receptor types in the chick retina. The distributions of unlabelled and labelled principal cones in maps of the receptor mosaic, constructed from retinae exposed to tritiated thymidine during incubation, were analysed by grid analysis (Greig-Smith, '64) and by two computer programmes. The computer programmes were generally more successful than grid analysis in detecting contagion in the distribution of unlabelled and labelled principal cones in the receptor maps. The contagion has been interpreted as evidence that the stage of development varies systematically in small retinal areas and that the overlap between the times of formation of the receptor types may be absent amongst small groups of cells.


Olfactory discrimination, reversal learning, and stimulus control in rats. Rats were trained to discriminate lights, tones, or odors and then given a series of discrimination reversals. Only rats trained with odors showed positive transfer on the first reversal and acquisition of a reversal set. Other experiments demonstrated that rats preferentially attend to odors when presented in compound with lights or tones; that odors exert more discriminative control than tones in tests using compound stimuli of competing sign; and that after pretraining on the positive stimulus, acquisition of an odor but not a light discrimination occurs with virtually no errors. These results demonstrate the importance of stimulus modality in the establishment of stimulus control and the need for more careful analysis of stimulus factors in cross-species comparisons of learning ability.


A rat's first bite: the nongenetic, cross-generational transfer of information. Weanling rats, when first exposed to solid food, showed relatively elevated intake if offered the same diet as eaten by their foster dams. Maternal caretakers pass some specific food-related information to their offspring.


Motivational and neuroanatomical specificity of hypodipsia for aversive solutions produced by medial preoptic injury to the rat. Bilateral medial preoptic lesions in rats dramatically lowered the rejection threshold for quinine-adulterated water but not for food in 24-hr forced-choice tests. The detection threshold for quinine in a two-bottle choice test, however, was unaffected by the medial preoptic lesion. Bilateral septal and lateral preoptic lesions had no effect on any quinine-adulteration tests. The enhanced rejection of quinine-adulterated water in a forced-choice test by medial-preoptic-damaged rats was also observed after 24-hr of water deprivation. Furthermore, the plasma osmolality of medial preoptic rats was significantly elevated above controls after 24-hr of water deprivation. These findings were interpreted as suggesting that a medial preoptic lesion produces a deficit in thirst-motivated behavior.


Failure of sodium- and calcium-deficient rats to acquire conditioned taste aversions to the object of their specific hunger. Attempts were made to condition taste aversions to the objects of two mineral-specific hungers. Both the innate preference of adrenalectomized rats for sodium and the learned preference of parathyroidectomized rats for calcium were studied. None of the sodium-deficient rats poisoned after drinking sodium chloride (NaCl) reached a taste-avoidance criterion, even after nine pairings of salt ingestion with aversive lithium chloride injections. Six of 11 calcium-deficient rats did not meet the salt-avoidance criterion after 10 pairings. Nondeficient control subjects learned to avoid these salt solutions completely after an average of only three such pairings. Besides unmasking a surprising degree of similarity between the learned and innate specific hungers studied, the results clearly demonstrate a powerful influence of physiological need on aversion conditioning.


Alcohol-induced conditioned aversion: genotypie specificity in mice (Mus musculus). Acetaldehyde poisoning from ethanol ingestion may lead to aversion to ethanol among DBA mice but not among C57s, since the former are relatively deficient in aldehyde dehydrogenase activity. The present study paired ingestion of saccharin with a single intraperitoneal injection of one of four concentrations of ethanol for DBA/2J and C57BL/6J mice. Subjects were then given a two-bottle saccharin versus water preference test for 10 days. Substitution of saccharin for the taste of ethanol resulted in avoidance of saccharin with all concentrations of ethanol by DBAs but not by C57s, consistent with the conditioned taste aversion paradigm as a model for genetically mediated ethanol avoidance.


Modification of the punishing effects of psychoactive drugs in rats by previous drug experience. In Experiment 1, it was shown that chronic prior exposure to amphetamine attenuated the conditioned avoidance of saccharin that was produced by both amphetamine and morphine during gustatory conditioning trials; the relationship between morphine and amphetamine was somewhat anomalous because of their pharmacological dissimilarity. The relationship was also asymmetrical, since in Experiment 2, chronic prior exposure to morphine failed to mitigate avoidance conditioning by amphetamine but was effective in attenuating conditioning by morphine itself. In a third experiment, it was found that prior treatment with chlordiazepoxide attenuated saccharin avoidance conditioned by chlordiazepoxide but not by amphetamine or morphine. The findings were related to Parker, Failor, and Weidman's hypothesis, based on findings with morphine, concerning the development of conditioned preferences for substances associated with the repletion of artificially induced biological needs. It was suggested that the findings were best interpreted as a reflection of drug tolerance rather than conditioned preference.


Physiological patterns accompanying complex problem solving during warning and nonwarning conditions. Physiological responses of 48 subjects were studied during both a warning and a nonwarning condition and during anagram problem solving. The warning condition was associated with heart rate deceleration and increased occurrence of galvanic skin responses (GSR), of skin conductance, and of tonic galvanic skin potentials (GSP). The warning condition also resulted in significantly faster solution rates. Problem solving was associated with increases in heart rate, occurrence of GSR, and skin conductance. The occurrence of phasic GSP habituated, but the occurrence of tonic GSP maintained a significant response frequency. Large increases in tonic GSP, with minimal phasic increases during problem solving, reflected the bidimensionality of electrodermal activity as a function of the task. These findings were interpreted to support Lacey's hypothesis of response fractionation and suggest the utility of physiological measures in the study of putative attentional functioning.


Skin potential activity in rats, cats, and primates (including man): a phylogenetic point of view. With rats, cats, nonhuman primates, and humans serving as subjects, skin potential activity was measured under waking conditions. In good agreement with the findings of previous workers, skin potential response waveform was always monophasic negative in rats and cats, but in humans it took three forms. By contrast, it was always monophasic positive in simian nonhuman primates, although prosimiae gave monophasic negative waves. A skin potential level-skin potential response relationship could not be observed in any subject except humans. From these results, an attempt was made to relate skin potential activity to the peripheral mechanism involved in these species on the basis of a phylogenetic point of view.


Correlation of overt escape behavior, multiunit thalamic activity, and midbrain lemniscal stimulation in rats. An acute and a chronic experiment were conducted in order to assess the extent to which ventral thalamic multiunit activity could account for the specific form of bar-pressing escape behavior in rats stimulated by trains of midbrain medial lemniscal pulse pairs. A paradigm used by Kestenbaum, Deutsch, and Coons was utilized in which the intra-pair interval of the train was varied. Results in anesthetized and freely moving animals indicated that midbrain lemniscal stimulation produces both an excitatory short-latency thalamic response showing the property of temporal facilitation and a long-lasting inhibitory process consistent with results of studies using anesthetized cats. The overall electrophysiological response, however, correlated significantly with the behavioral response function.


Laboratory observations of trailing behavior in garter snakes. Using a Y-maze apparatus, nine garter snakes were trained to follow an earthworm-extract trail. Correct trailing improved significantly as training progressed. All snakes were able to attain a trailing criterion of 8 out of 10 correct trials by the termination of training. For each trial, the number of tongue flicks each snake made was recorded and tongue flick rate calculated. Tongue flick rate was found to be highly characteristic and constant for individual subjects. In addition, running speed and tongue flick rate were found to be significantly correlated.


Development of species identification in ducklings: II. Experimental prevention of perceptual deficit caused by embryonic auditory deprivation. Devocalized-isolated ducklings are relatively insensitive to the higher frequencies in their species' maternal call 24 hr after hatching. To assess the efficacy of various sounds in preventing this high-frequency perceptual deficit, devocalized-isolated embryos were exposed to one of three calls and tested with normal versus high-frequency attenuated maternal calls 24 hr after hatching. The embryonic vocalization (contact-contentment call) that most closely matched the region of the devocalized ducklings' greatest insensitivity (1,500-2,500 Hz) proved most efficacious in remedying (preventing) the high-frequency perceptual deficit. Exposure to the embryonic alarm-distress call was somewhat effective, whereas exposure to low-frequency white noise was not effective at all. This finding suggests that the normal development of innate behavior is partially dependent upon prior experience as well as upon intrinsic processes of neural maturation. Although normally occurring embryonic auditory experience does not induce the preference for the maternal call in this species, such experience does contribute to the sharpness of the discriminative basis of the preference 24 hr after hatching.


Hysterectomy-induced maternal behavior during pregnancy in the rat. Hysterectomies were performed between the 10th and 19th day of pregnancy, and females were tested with pups for the onset of maternal behavior starting 0,24, 48, or 72 hr after surgery. Pups remained with the females overnight, and testing was repeated daily with fresh pups until females exhibited maternal behavior. Latencies for the onset of maternal behavior were shorter after hysterectomy on the 10th and 16th days of pregnancy than in intact pregnant females at the same stages of pregnancy; latencies became shorter, the later the termination of pregnancy. When the ovaries were removed along with hysterectomy during pregnancy, short-latency maternal behavior no longer was exhibited. Pregnant females were tested during the last 40 hr of pregnancy: nest building began at 34 hr prepartum and retrieving at 28 hr prepartum. The effect of hysterectomy during pregnancy on ovarian secretion of estrogen and progesterone is reviewed, and it is concluded that the rise in estrogen secretion, which follows hysterectomy during pregnancy, is most likely the cause of the rapid onset of maternal behavior after hysterectomy. A similar proposal is made for the prepartum onset of maternal behavior in intact pregnant females.


Olfactory basis for the delayed onset of maternal behavior in virgin female rats: experimential effects. A previous study showed that intranasal zinc sulfate (ZnSO4) treatment shortens the latencies for the onset of maternal behavior in nonpregnant rats. The aim of this study was to see whether after recovery from the effects of ZnSO4 treatment, the latencies for maternal behavior increased. In Experiment 1 nonpregnant females were intranasally infused with ZnSO4 or air or were left untreated and exposed to pups starting 48 hr later. Olfactory discrimination tests using chocolate bits were done simultaneously. The ZnSO4 treatment resulted in short latency maternal behavior (1.1 days) compared with latencies of control females (3.8 days), but neither group showed any loss of olfactory discrimination. Three weeks later- ZnSO4-treated females showed increased latencies for all but retrieving, while control groups showed decreased latencies for all maternal behaviors. In Experiment 2, intranasal ZnSO4- and air-treated females were given olfactory discrimination tests under food deprivation, using chocolate bits and guinea pigs pellets. Choice of guinea pig pellets was more severely affected than choice of chocolate bits, but recovery of the discriminations was complete in 4-5 days. The effects of ZnSO4 on maternal behavior and on olfactory discrimination in nonpregnant females were discussed.


Ontogenetic dissociation of two classes of habituation. The rates of decrement of two classes of response (an elicited startle reflex and emitted exploratory behavior) were determined in rats of two different ages (15 and 36 days). The rate of decrement in the startle reflex was not clearly differentiated as a function of age. In contrast, there was no evidence of habituation of exploration in the younger animals, whereas the older rats uniformly showed profound response decrements. This ontogenetic dissociation of the two instances of response decrement indicates that accounts of both instances in terms of a common process called habituation may be unwarranted. In addition, these data, in conjunction with earlier findings, indirectly support the possibility that reflex decrements may be relatively more dependent on brain serotonin whereas decrements in exploration may be more dependent on normal cholinergic activity in brain.


Ontogeny of thermoregulatory mechanisms in the rat. The ability of rats to select a warm environment was studied as a function of postnatal age (birth to 13 days). Animals younger than 5 days demonstrated no choice response (movement to a warm compartment, 36-37 degrees C); however, they did demonstrate movement within the start compartment (23 degrees C). Increasing the motor capabilities of the pups, by injection of L-3,4 dihydroxyphenylalanine (L-dopa, 50 mg/kg), elicited a choice response in 4-5 day-old animals. Younger animals demonstrated no choice of a warm environment even though they moved considerably. Also there was no difference between L-dopa-treated and control animals in the magnitude of temperature change in pups isolated from their mother for 1 hr. The evidence suggests development of behavioral thermoregulatory mechanisms prior to abilities for internal regulation.


Persistent depression of rearing behavior in rats after extensive septal lesions. Extensive septal lesions produced a persistent (76 days) reduction in rearing behavior which was not directly attributable to altered deprivation states, emotionality, activity levels, or competing behaviors and which occurred in familiar as well as various novel testing environments. Changes in visual, olfactory, or conspecific stimuli ameliorated or had no effect on the lesion effect.  The results indicate that the septal area is importantly involved in normal rearing behavior and that lesion-induced alterations in responsiveness to some forms of stimulation may play a modulatory role in reduced rearing. It is also suggested that the septal area may constitute a portion of a neural system responsible for the initiation of this form of response.


Locomotor, avoidance,and maze behavior in rats with selective disruption of hippocampal output. Behavioral correlates of selective disruption of hippocampal output were investigated in a series of five experiments. In two experiments an attempt was made through behavioral investigation to determine whether the CA1 neurons project to the fimbria or to the subiculum. The results supported recent views that the subiculum is the recipient of CA1 axons. Disruption of the CA1 output in the dorsal hippocampus of rats produced increased open-field activity, whereas passive avoidance and spontaneous alternation behaviors remained unchanged. No differentiation was obtained between CA1 damage and neocortical lesions in maze learning. Blocking of the fimbrial CA3 output from the dorsal hippocampus improved passive avoidance performance and impaired active avoidance performance, whereas open-field and spontaneous alternation behaviors were unaffected. Interruption of the CA3 output from the ventral hippocampus improved active avoidance performance and reduced spontaneous alternation behavior. Open-field behavior and passive avoidance performance remained unchanged. Total fimbrial sections increased open-field activity, improved passive and active avoidance, and reduced spontaneous alternation. The results are discussed in terms of functional differentiation between the CA1 and CA3 of the dorsal hippocampus and in terms of functional differences in the fimbrial CA3 output from the dorsal and ventral hippocampus.


Intrameal eating patterns of obese and nonobese humans. Two experiments were conducted to test whether the intrameal eating patterns of obese and normal-weight human subjects are different. The first experiment was conducted in a laboratory under the pretense of a taste test. Eating was observed and data were compiled for seven parameters related to eating. The second experiment was conducted under field-study conditions with the subjects unaware of their participation. Few statistically significant differences in eating behavior were observed between obese and normal-weight subjects. These results are in contrast to the majority of previous work.


Cholecystokinin elicits the complete behavioral sequence of satiety in rats. The behavior of intact rats and rats with chronic gastric fistulas was observed and scored during a 60-min test period when they were offered liquid diet after 17 hr of food deprivation. Intact rats and rats with closed fistulas displayed a specific behavioral sequence at the end of each meal: They stopped eating, engaged in grooming and exploration for a short time, and then rested or slept. Thus, a fixed behavioral sequence characterizes satiety in the rat. Although the behavioral sequence of satiety was fixed, the cessation of feeding was not a sufficient condition for the appearance of the rest of the sequence: Quinine adulteration of the liquid diet stopped sham feeding but did not elicit the complete sequence. Intraperitoneal injection of the intestinal hormone cholecystokinin during sham feeding, however, elicited the complete sequence of satiety. The observation that cholecystokinin not only stops feeding but elicits the complete sequence of satiety supports our hypothesis that endogenous cholecystokinin is a satiety signal for the rat.


Ontogeny of meal patterning in rats and its recapitulation during recovery from lateral hypothalamic lesions. The development of feeding patterns was investigated in weanling rats and in rats with lateral hypothalamic lesions. From 16 to 25 days of age, the weanlings demonstrated a preprandial intake pattern, i.e., a positive correlation between meal size and time since the preceding meal. This subsequently declined while the postprandial relationship (correlation with time until subsequent meal) began to emerge such that by 30-35 days of age a full adult pattern was observed. Rats recovering from lateral hypothalamic lesions, for a brief period, also demonstrated a preprandial intake pattern. The postprandial relationship was abolished by the lesions. These results suggest that the development of adult meal patterning results from maturation of lateral hypothalamic mechanisms governing meal initiation.


Development of IgE and allergy in infancy. Serum IgE levels were followed throughout the first year of life in a sample of 34 infants from atopic and nonatopic families. Their parents' serum IgE levels were also measured. Neonatal levels ranged from 0 to 10 U/ml. Half of the study group, which included a high proportion of infants from atopic families, maintained serum IgE levels below 10 U/ml until 1 yr of age. In the remainder, the serum IgE exceeded 10 U/ml before 1 yr of age, but the time of increase and the rate of increase varied from infant to infant; the sooner the IgE started to increase, the higher was the serum IgE level at 1 yr of age. The relationship of serum IgE level to the development of atopic disease and to other parameters was analyzed. Elevation of serum IgE level at or before 1 yr of age was highly correlated with atopic disease in the first 2 yr of life. The elevation preceded the manifestation of atopy.


A comparison of methacholine and histamine inhalations in asthmatics. Bronchial provocation tests using aerosolized serially diluted histamine and methacholine were given to nearly 200 asthmatics. Results were usually reproducible for a given patient and corticosteroids did not influence the procedures. Those patients who could tolerate high doses of methacholine were statistically the least severe asthmatics as measured by their discharge dose of corticosteroids. Reaginmediated asthmatics could not tolerate the higher doses of histamine. These tests help delineate subgroups of asthmatics and may have clinical usefulness since, when combined with other data, they differentiate pathogenetic mechanisms in some patients and suggest therapeutic approaches in others.


SECA, a new mediator of the human eosinophil response. Spontaneous eosinophilic chemotactic activity (SECA) present in human sera can mediate the directed movement of normal human eosinophils. Our data utilize normal peripheral blood eosinophils obtained from subjects with 500 eosinophils/m3 or less. SECA is defined as that chemotactic activity for eosinophils present in serum that has been heat-inactivated immediately after collection. It was demonstrated in patients with severe chronic eczema with eosinophilia (20 to 30%); mixed collagen vascular disease with vasculitis; clinical serum sickness; acute glomerulonephritis, and chronic membranoproliferative glomerulonephritis. Control sera were obtained from normal, healthy individuals. The data indicated: (1) that SECA in patient sera was significantly higher than in control sera; (2) when activated by endotoxin, no additional chemotactic activity was generated from patient sera over that spontaneously present--by contrast, addition of endotoxin to control sera did result in increased chemotactic activity; (3) sera from patients with extrinsic bronchial asthma had no SECA.


Hospital dietetics in transition.

Hospital dietetics in transition. Nutritional care--what is it, who gets it, who gives it, and how? The registered dietitian is identified as the only health professional whose primary concern is the application of food and nutritional science to the health care of people. Traditionally, the major part of health care has been in the medical sphere--sought in times of acute illness or injury. Hospital-based services provide increasingly sophisticated life support systems in specialized intensive care units. Humane, as well as financial, considerations mandate a change in emphasis to the provision of preventive care as well as restorative measures. Care that provides nutritional status screening, dietary assessment, and dietary counseling is seen as essential for health maintenance.... The facts of food composition are not enough!


Dietitians' attitudes toward advocacy.

Dietitians' attitudes toward advocacy. A survey was conducted to determine dietitians' attitudes toward and readiness for advocacy. Representative members of an affiliated state dietetic association expressed positive attitudes toward the professional's assumption of the advocate role. The respondents looked to the national, state, and local dietetic associations to provide leadership in the area of advocacy.


Attachment learning: a humanistic educational experience. Final practicum in a coordinated undergraduate program. Throughout the five stages, the student has ample opportunity for expression and self-evaluation in the counseling sessions that accompany each stage. The student experiences progress with increasing responsibility to the ultimate functioning as an administrative dietitian under supervision. It is believed that the experiences of attachment learning will enable the student, on graduation, to enter professional practice with confidence and a strong likelihood of success. Thus far, experiences with attachment learning at the University of Missouri-Columbia have demonstrated superiority over methods used previously. It is believed that this method can be readily adapted to other learning situations in dietetics, such as traineeships and internships. In fact, our enthusiasm for the method prompts the suggestion that it might well be considered for the final stage of education for any profession in which entry-level competencies can be established.


The public seminar as a nutrition education approach. An evaluation. A health seminar seemed to modify knowledge, attitudes, and reported behavior of a self-selected audience which was already well informed and motivated to change. Therefore, the health seminar appears to be a useful educational approach in this context. The question remains: How can those who are neither informed nor motivated be reached? This study suggests that the public seminar may not be an efficient vehicle for this purpose.


Urinary steroids in the measurement of aging and of atherosclerosis. Several studies conducted by the authors' group have shown that urinary steroid measurements are a valuable aid in differentiating the normal aging process, the pronounced aging associated with increased risk to coronary heart disease, and the deviations associated with myocardial infarction. Data are presented on 428 men in the age range of 30-70 years. The study design most effective in elucidating aging and disease patterns involves selection of subjects from a wide age range. Data on persons identified as clinically normal can be used to describe physiologic aging. Once this is determined, data on persons with disease can be used to identify abnormalities of aging associated with the clinical conditions studied. This approach offers a potential method for differentiating between aging effects and disease effects. The foregoing findings led to the development of an Index of Aging in males, based on combined serum lipid and urinary steroid values. This Index may be a means of differentiating between normal aging and the deviations seen in atherosclerosis and myocardial infarction. Current studies are directed toward extending these observations.


Glucocorticoid therapy in sepsis/shock caused by gram-negative microorganisms. A review of the literature reveals that glucocorticoids have: 1) a protective effect both in vitro and in vivo against bacterial endotoxins and exotoxins, and 2) a protective or therapeutic effect in sepsis/shock caused by Gram-negative microorganisms. At the Columbia Presbyterian Medical Center, the 1968-1973 mortality rate for general sepsis/shock due to Gram-negative organisms was 16.6 percent and for urologic sepsis/shock 15.3 percent. This low rate was the result of early diagnosis and early use of massive dosages of glucocorticoids (hydrocortisone, methylprednisolone, dexamethasone), beta-adrenergic isoproterenol, and bactericidal antibiotics (gentamicin, kanamycin, carbenicillin, cephalothin-cefazolin, ampicillin).


Lipidemia--a multigeneration predictability study. This study involved a 1st generation of 194 aged residents at the Philadelphia Geriatric Center plus 2nd and 3rd generations of 357 and 306 offspring, respectively. Serum cholesterol levels (including type of abnormality) and triglyceride levels were determined. The results suggest that hyperlipidemia at all ages is related to a familial genetic problem. The data on each generation, starting even with the 80-year-old parents, could be used to predict lipid abnormalities in each succeeding generations. However, when comparing the 1st generation with the 3rd generation, predictability was lacking, probably because of the inclusion of two sets of spouses and many enviromental factors that altered the situation. In the 2nd generation, 40 per cent of the subjects had an elevated lipid level compared to 30 per cent in the 1st and 3rd generations. The higher concentrations in the 2nd generation probably reflected increased risk factors such as improper diets, smoking, hypertension, obesity and stress at that age level. Hyperlipidemia in the aged apparently is not just a metabolic degenerative abnormality. It should be treated, as in younger people. Data on risk factors such as a high blood lipid level may help not only the aged, but succeeding generations.


Hepatic infarction associated with terminal hemorrhagic necrotizing enteropathy (THNE): case report. A case of hepatic infarction associated with terminal hemorrhagic necrotizing enteropathy (THNE) in an 84-year-old woman with cardiac failure is presented. The autopsy showed hepatic infarction, THNE, fibrous endocarditis, generalized arteriosclerosis, myocardial hypertrophy, liver congestion, and other findings as described. Both rare infarctions (hepatic and intestinal) seemed to have originated not only as a result of impaired systemic circulation due to cardiac failure, but also from liver congestion due to peripheral circulatory deficiency through the portal vein. The circulatory correlation between these infarcts of the liver and intestine is discussed.


Pelvic distress in the aged patient.

Pelvic distress in the aged patient. Unexplained pelvic distress or pain was observed in the 4 patients of this report (3 women and 1 man), and also in 6 other patients during a 10-year period of neurologic practice. In each of the 4 reported cases, the discomfort was interpreted as sexual. It is suggested that this may be a valid entity and that, as with trigeminal neuralgia and postherpetic neuralgia, it is truly limited to aged patients. The mechanism, incidence and significance are unknown.


A psychogeriatric assessment program. II. Clinical and laboratory findings. In a Psycholgeriatric Assessment Program, the statistical evaluation of clinical and laboratory data related to thyroid function in 80 patients led to the conclusion that: 1) differences in thyroid function between psychogeriatric patients of the organic and functional types were only slight, as determined by clinical symptoms and thyroxine blood levels, and 2) factor analysis for 53 selected variables indicated that loading for "gross disturbance of psychobiologic function" may be related to the symptom complex of hyperthyroidism/hyperadrenergism, and particularly to psychobiologic symptoms rather than purely somatic symptoms of hyperthyroidism. In comparison with a group of 60 healthy old people, the psychogeriatric patients showed many more signs of hyperactivity and other characteristics of this symptom complex. The results supported the preliminary impression of an energetic (hyperthyroid-sympathicotonic) reaction to mental decline, accompanied by plasma corticosteroid and electroencephalographic changes.


A modification of the unlabeled antibody enzyme method using heterologous antisera for the light microscopic and ultrastructural localization of insulin, glucagon and growth hormone. The requirement of using homologous antisera (primary antiserum and peroxidase-antiperoxidase (PAP) complex raised in the same species) in the unlabeled antibody enzyme method has been investigated at the light and electron microscopic level using the localization of insulin, glucagon and growth hormone as model systems. Optimum immunocytochemical staining for all three antigens was observed when sheep or goat antirabbit gamma-globulin (S-ARgammaG or G-ARgammaG) were used to couple rabbit peroxidase-antiperoxidase complex with either guinea pig antisera to insulin (GP-AIS) or glucagon (GP-AGS), or monkey antisera to rat growth hormone (M-ARGH). The cross-reactivity between S-ARgammaG or G-ARgammaG and immunoglobulins in these primary antisera were substantiated by immunoelectrophoresis and radioimmunoassay. S-ARgammaG was shown to produce precipitation arcs with GP-AIS and M-ARGH that were similar to those seen when the latter were reacted with rabbit antiguinea pig gamma-globulin antiserum and goat antimonkey gamma-globulin antiserum, respectively. Radioimmunoassay results revealed that immunoprecipitation of 6-10% as compared to homologous antisera controls yielded excellent staining localization when S-ARgammaG was used for immunocytochemistry. Thus, heterologous antisera (primary antiserum and PAP complex raised in different species) may be used in the unlabeled antibody enzyme method as long as the coupling antiserum shows cross-reactivity with immunoglobulins of the primary antiserum and the PAP complex.


The efficiency of 14C detection in autoradiographs of semithin plastic sections. Sections of uniformly labeled 14C-methylmethacrylate were used to determine the efficiency of 14C detection by Ilford L4 emulsion layers, 0.22 mu in thickness, in light microscope autoradiographs. When efficiency was expressed as a function of radiation dose (i.e., the number of decays/mu2) there was a strong dependency on radiation dose up to about 6 decays/mu2, after which detection efficiency was fairly constant. For most emulsion batches, the detection efficiency was about 5%, but it ranged from 8 to 3%.


Intranuclear silicon detection in a subcutaneous connective tissue cell by energy-dispersive x-ray miscroanalysis using fresh air-dried spread. Silicon was detected by energy-dispersive x-ray microanalysis in the nucleus of a subcutaneous connective tissue cell of mice fed normally. To eliminate contamination, pieces of connective tissue were spread on copper grids and examined without any treatment by an energy-dispersive spectrometer with a scanning transmission apparatus attached to an electron microscope. Scanning transmission electron microscopy of the spread has demonstrated a well-preserved ultrastructure. Fibrous structures, nuclei and nucleoli of cells and mitochondrial granules were recognized. Electron probe analysis showed peaks for silicon at three spots on the nucleus of a cell in addition to those for phosphorus, sulfur, chlorine, potassium and calcium, whereas no peak of silicon could be detected at the spots on nuclei of other cells, mitochondrial granules and electron-lucent area on the same grid as the above. Silicon appears to play a significant role in the nucleus. Applicability of the technique to know the distribution of easily contaminating elements and diffusible substances is shown.


Modification toward dominance of a recessive lethal in the mouse. The effect of the recessive siren (srn) gene singly in the presence of one Danforth's short tail (Sd) gene was to modify the expression of srn towards dominance with 46 percent penetrance. All mice that could be classified as sirens from srn/+ x Sd/+ matings had the Sd type short tail, indicating that interaction between Sd and srn was the principal cause for modification towards dominance of srn. External expressivity was variable in Sd srn pups, with true sirens, semi-sirens and apposed leg pups produced, each traceable to pelvic abnormalities. Variation in the urogenital system was similar to that found in siren pups with the srn/srn genotype.


Subfragments of the papain solubilized TL antigen. TL antigen was solubilized from the tumor ASLI (TL. 1,2,3) by papain digestion. The subfragments of 125I-labeled TL were examined by two methods. The first involved immune precipitation followed by electrophoresis on SDS-acrylamide gels. This treatment yielded three bands of molecular weight 39,000 and 19,000, as well as material which migrated with the tracking dye. In the second procedure the papain digested material was partially purified on Sephadex G-200. The active fraction from G-200 was labeled with 125Iodine, mixed with alloantiserum and rechromatographed on G-200. The isolated immune complexes were boiled in SDS and 2-mercaptoethanol, then separated on a SDS-Sephadex G-150 column. Two radioactive peaks were eluted indicating an absence of the 19,000 m.w. component following the latter method of purification.


Genetics of house flies. Variability studies with North Dakota, Texas, and Florida populations. Genetic data were used to compare the structure of native house fly populations collected in North Dakota, Texas, and Florida. Recombination studies with mutant markers on chromosomes 3 and 4 indicated a lack of inversion polymorphism among the three populations in those areas of the genetic map studied. Significant differences were observed among flies from the three regions with regard to the frequency of 1) females that produced only male progeny, and 2) male-determining 3rd chromosomes (IIIm chromosomes). However, the North Dakota and Texas flies were more similar to each other than to the Florida flies since populations from the two former areas possessed a low frequency of both male-producing females and IIIm chromosomes; in contrast, the Florida population was void of females that produced males only and a high percentage if not all Florida males appeared to possess the IIIm male-determining mechanism. Tests for recessive lethal 3rd chromosomes showed that there was no significant difference in the frequency of lethal factors recovered from the North Dakota and Texas flies; the presence of IIIm chromosomes in Florida males precluded the recovery of lethal factors from this population by the method employed. The data suggest that house fly strains to be employed in genetic control programs should 1) originate from target control areas to avoid possible behavioral differences existing among flies from different locales, 2) be initiated with as many flies as possible to provide a background for the maintenance of variability, and 3) be renewed periodically with field-collected material since the genotype may be capable of rapid reorganization in response to laboratory selection pressures.


Stimulation of human lymphocytes by galactose-specific abrus and ricinus lectins. Human lymphocyte cultures were incubated with the nontoxic abrus agglutinin and with ricin B chain, and the incorporation of 3H thymidine was measured. Abrus agglutinin stimulated strongly the thymidine incorporation whereas ricin B chain had a much lesser effect. When galactose or lactose was added to the cultures together with the lectins, the abrus agglutinin and ricin B chain induced thymidine incorporation was strongly reduced. There was a linear relationship between the concentration of lectin and the concentration of lactose required for inhibition of lymphocyte stimulation. N-acetyl-galactosamine had a much lesser inhibiting effect and alpha-methyl-mannoside did not cause any inhibition. The abrus agglutinin induced thymidine incorporation was not demonstrable before 36 to 40 hr and reached its maximum after 2 to 5 days. If lactose was added within the first 4 hr of incubation with abrus agglutinin no stimulation was observed.


Complement inhibitor(s) released by leukocytes. I. Pretreatment of sheep erythrocytes with supernatants of mouse spleen and thymus cells inhibit whole complement activity and C2 utilization. Sheep erythrocytes pretreated with supernatants of mouse spleen or thymus cells become resistant to lysis by guinea pig complement. The inhibitory activity (IA) reduces the utilization of C2 by EAC14. Because IA binds to the surface of sheep erythrocytes and does not inhibit C1 irreversibly, it is probably a hitherto undescribed inhibitor of complement.


Complement inhibitor(s) released by leukocytes. III. Evidence for a "new" C1 inhibitor in the supernatants of short-term cultures of mouse spleen and thymus cells. Mouse spleen or thymus cells in short-term culture release a factor, designated S, that binds to sheep erythrocytes (E). Supernatant-treated sheep erythrocytes (SE) are capable of fixing and transferring the activated first component of guinea pig complement. SEC1, however is not capable of initiating hemolysis by the rest of the complement components. SE is capable of binding but not activating native C1; native C1 bound to SE seems irreversibly inhibited. Evidence is presented that S may be the same factor as the previously described inhibitor released by mouse spleen or thymus cells that inhibits the utilization of C2 by EAC14.


Biologic properties of transplantation immune sera. IV. Influence of the course of immunization, dilution and complexing to antigen on enhancing activity of Ig classes. The influence of the course of immunization on the facilitating-enhancing activity of antibody classes has been studied by passive enhancement of growth of A/JAX sarcomas in CBA and IC mice and of C57BL/6 EL 4 leukemia in BALB/c mice. The influence of dilution of antibodies and complexing to antigens was also studied. During immunization (with several boosters), the enhancing capacity of sera increased together with 7S IgG antibody activity, but showed no correlation with 19S IgM antibody activity. It also was mercaptoethanol resistant. IgG1 to be more enhancing than an equal number of hemagglutinating units of IgG2a. When concentrated on a small amount (10(5)) of target sarcoma I cells, complement-fixing IC anti-A antibodies were even inhibitory on Sa I allografted to IC recipients. Progressive dilutions reversed this situation, IgG1 activity disappearing and IgG2 acquiring enhancing activity. After complexing to corresponding antigens IgG2 also (and immune sera with inhibitory properties) acquired enhancing properties. These results may provide a basis for understanding the discrepancies between the results of several groups of authors studying the class(es) of enhancing anibodies.