Ultrastructure of mitosis in the chloromonadophycean alga Vacuolaria virescens. During preprophase in the chloromonadophycean alga Vacuolaria virescens microtubules are present around the flagellar basal bodies and extend over the anterior surface of the nucleus. These microtubules assist in the separation of the flagella and later enter the nucleus through polar gaps. During prophase the nucleoli begin to disperse and the chromosomes become condensed. At metaphase the nucleus assumes an elliptical shape and an equatorial plate of chromosomes becomes aligned across the long axis of the nucleus; kinetochores are recognizable on some of the chromosomes. The nuclear envelope remains intact over most of the surface and in places it forms folds. During anaphase chromosomes are less distinct and vesicles are present in the elongating nucleus. Most of the new nuclear envelope around the progeny nuclei is formed by coalescence of these membrane vesicles during late anaphase and telophase, although some of the original nuclear envelope may also become incorporated. During telophase disintegration of the original nuclear envelope becomes pronounced and portions of this structure are recognizable in the cytoplasm after completion of mitosis. It is suggested that this unusual type of nuclear envelope behaviour may be important in ensuring the segregation of the Golgi apparatus and contractile vacuole to progeny cells. Interphase cells contain a single extensive Golgi apparatus which is located between the anterior surface of the nucleus and the contractile vacuole. The Golgi apparatus and contractile vacuole act as an osmoregulatory system and their presence is presumably essential to the existence of the organism. Formation of a new contractile vacuole and division of the Golgi apparatus occur early in mitosis and thereafter a Golgi apparatus and contractile vacuole become associated with each of the poles of the nucleus. They retain this location throughout mitosis and during cytokinesis, with the result that an osmoregulatory system is present in each of the daughter cells. In a similar manner, microbody-like organelles are associated with the nuclear envelope during mitosis but not at interphase. Growth of the nuclear envelope during mitosis may serve as the means of partitioning these organelles to the progeny cells. Thus mitosis in Vacuolaria virescens is responsible not only for the equal segregation of the genetic material but also for the correct distribution of some of the cytoplasmic components.


The arrangement of microtubules in serially sectioned spindles of the alga Cryptomonas. Serial sections of mitotic spindles of the marine cryptophycean alga, Cryptomonas, were analysed to determine what types of microtubules they contained and which of these microtubules came close enough to each other (50 nm or less) for the commonly described crossbridging to be possible. Interpolar microtubules were rare (less than or equal to 1%) but from prometaphase through anaphase there was a substantial interpolar framework of free and polar microtubules which came close enough to one another to cross-bridge and generate anaphase spindle elongation by intermicrotubule sliding. However, such elongation would also require some concomitant polar microtubule polymerization. In contrast, only about 12% of the chromosomal microtubules came within bridging distance of interpolar framework microtubules. Thus, assuming that microtubules were accurately fixed in their in vivo positions, crossbridging between chromosomal and interpolar framework microtubules is unlikely to function in chromosome-to-pole movement. In all stages the great majority of chromosomal microtubules were found to extend all the way from the chromatin to the pole. Although the identification of intermicrotubular bridges in section is often ambiguous, the best and most frequent examples of bridges in the present material were between polar microtubules and between chromosomal microtubules but not between the 2 different categories. The spindles also contained 5-nm-diameter microfilaments associated mainly with chromosomal microtubules and occasionally with polar microtubules. A specific model for the possible involvement of these filaments in mitosis is presented.


Ultrastructure and activity of the nucleolar organizer in the mouse oocyte during meiotic prophase. The mouse oocyte is the site of nucleolar synthesis during pachytene. The chromosomes containing a nucleolar organizer are attached to the nuclear envelope by their paracentromeric heterochromatin, either alone or by taking part in the formation of a chromocentre. The nucleolus appears at the junction of the paracentromeric heterochromatin with the euchromatic portion of the bivalent. In this zone, 5.0-nm-diameter fibres, thinner than those of the rest of the chromosome (10.0 nm), extend from the lateral element of the synaptonemal complex up to the nucleolar fibrillar centre in which they penetrate. At the onset of its synthesis, the nucleolus only contains the fibrillar centre and an electron-dense fibrillar component in continuity with the latter. Growth of the nucleolus often takes place in the form of a strand whose proximal end, in contact with the fibrillar centre, is formed by preribosomal fibrils and whose distal end is at first fibrillo-granular then granular. Following brief incorporation of tritiated uridine, nucleolar labelling is active in oogonia. No ribosomal RNA-synthetic activity is revealed during leptotene and zygotene. Incorporation resumes at mid-pachytene, with labelling located over the electron-dense fibrillar component adjacent to the fibrillar centre. These observations suggest that the rDNA is located in both the fibrillar centre and its associated electron-dense fibrillar component and that the rDNA transcription occurs in the latter.


Determination of trimethoprim and sulfamethoxazole (co-trimoxazole) in body fluids of man by means of high-performance liquid chromatography. A high-performance liquid chromatographic method for the determination of trimethoprim, sulfamethoxazole and its metabolite and a series of structurally related sulfonamides is described. The half-life time of elimination of sulfamethoxazole and its metabolite N4-acetylsulfamethoxazole is 9 h for both compounds. The renal excretion rate of sulfamethoxazole depends strongly on the urinary pH. The renal excretion rate of the metabolite N4-acetylsulfamethoxazole is not dependent on the urinary pH.


Quantitation of zimelidine and norzimelidine in plasma using high-performance liquid chromatography. A method is described for the quantitation of new non-tricyclic antidepressant, zimelidine, and its pharmacologically active, N-demethylated metabolite, norzimelidine, in plasma. The method involves a single extraction of basified plasma with diethyl ether, concentration of the ethereal extract, chromatography on a high-performance liquid chromatograph and quantitation using a variable-wavelength UV detector. The respective geometric isomers of zimelidene and norzimelidine are used as internal standards for quantitation. Resolution is effected using a 5-micron silica gel column with an aqueous methanolic solution of ammonium nitrate as the mobile phase. The minimum quantitated amount was 25 ng and the coefficient of variation for the method did not exceed 7% in the range 25 to 1000 ng/ml for both compounds. The method has been applied in monitoring the plasma concentration of zimelidine and norzimelidine in plasma from depressed patients and an example of this application is presented.


Determination of kynurenine by a simple gas-liquid chromatographic method applicable to urine, plasma, brain and cerebrospinal fluid. A simple, sensitive and specific method for the determination of kynurenine is described. This is based on alkaline cleavage of kynurenine, followed by solvent extraction, trifluoroacetylation and gas--liquid chromatography with electron capture detection. Using this method kynurenine has been determined in urine and plasma, and for the first time in brain and cerebrospinal fluid. Increases in kynurenine in brain, plasma and urine are demonstrated following tryptophan administration to man and rat.


Determination of urinary thiamine by high-pressure liquid chromatography utilizing the thiochrome fluorscent method. A sensitive, reproducible, and specific method for the determination of urinary thiamine has been established. Unique to the use of high-pressure liquid chromatography (HPLC) to separate the fluorescent thiamine derivative from interfering fluorescent compounds. Urine samples were passed through a Decalso catoin-exchange column, washed with 0.5 M KCl to remove some interfering compounds, and eluted with 3.4 M KCl. The eluted thiamine was converted to the fluorescent derivative, thiochrome, by reaction with alkaline potassium ferricyanide. The reaction mixture was extracted with isobutanol and subjected to HPLC monitored by a fluorescent detector. Within-day and day-to-day coefficients of variation proved to be 2.5% and 1.2% respectively. Recovery of added thiamine (range 0.04 to 2.0 microgram/ml) averaged 99.9 +/- 5.3%. The sensitivity of this method was 0.03 microgram/ml.


[Determination of "middle molecules" presenting vitamin B12 molecular size in normal and uremic body fluids (author's transl)]. Uremic solutes with the molecular size of vitamin B12 are assumed to be toxic. An analytical method is proposed to detect and separate these solutes in body fluids using two combined techniques: gel filtration on Sephadex G-15 and ion-exchange chromatography on DEAE-Sephadex A-25. The vitamin B12 molecular size has been localized by ultrafiltration through membranes with a defined cut-off. Normal and uremic body fluids (urine, plasma, hemodialysis fluid) have been separated into 9 ultraviolet-absorbing peaks (a to i) by high-speed gel filtration. Peaks b and c present the molecular size of vitamin B12, 10--15 A molecular diameter in pH7 aqueous solution. Peak b, which correlates with uremic neuropathy, is separated into 6 sub-peaks (b1 to b6) by ion-exchange chromatography, sub-peak b4.2 is the only one to correlate with uremic neuropathy. The coefficient of variation in the integrated area of a single peak is 16%. This method gives the chromatographic prolife of the vitamin B12 molecular size from 500 microliter of uremic plasma or 100 microliter of normal urine within one hour.


Monoacylcadaverines in the blood of schizophrenic patients. Concentrations of cadaverine, monoacetylcadaverine and monopropionylcadaverine in the blood of schizophrenic and nonschizophrenic subjects were measured. Two groups, one from the U.S.A. the other from Japan, were tested. Monoacetylcadaverine and monopropionylcadaverine were found elevated in the blood of some schizophrenic patients in comparison with those in controls in each group. Their increase could be caused by a reduced monoamine oxidase activity or by an increased acylation in schizophrenic patients.


Electrophoretic bimorphism of serum albumin in the presence of indocyanine green. The presence of indocyanine green during extended traditional electrophoresis and immunoelectrophoresis of serum is associated with bimorphism of albumin. This occurs over a range of dye--albumin molar ratios an order or more greater than was obtained in similar phenomena described previously. The bimorphism seems not to be dose dependent beyond a certain point, and the two albumins so separated show tinctorial differences. The phenomenon has been observed to apparently the same degree in all normal sera tested, and may represent a means of distinguishing ligand-loaded and ligand-light serum albumin.


Microdetermination of caffeine in blood by gas chromatography--mass spectrometry. A micromethod for the quantitative analysis of caffeine present in small quantities (100 microliter) of whole blood is described. It is based on the gas chromatographic--mass spectrometric analysis of chloroform extracts of biological samples. The method is relatively simple, rapid, specific and sensitive; as little as 20 ng of caffeine can be measured.


Paraffin oil pneumonia. Analysis of saturated hydrocarbons in different human tissue. Temperature-programmed gas chromatographic analysis on columns packed with Apiezon L as stationary phase is shown to be the best method for the qualitative and quantitative analysis of simple and complex hydrocarbon mixtures when compared with all the other applicable techniques (thin-layer chromatography, column chromatography, ultraviolet spectroscopy, infrared spectroscopy, nuclear magnetic resonance spectroscopy, mass spectrometry) described in this paper. Using the method in a patient with mineral oil pneumonia it could be demonstrated that he expectorated a maximum of 79.5 mg liquid paraffin daily and also transported equally complex saturated hydrocarbons in a concentration of 1.3 mg% in plasma and of 1.6 mg% in the cellular blood components. In an additional experiment the direct determination of liquid paraffin resorbed from the gastrointestinal tract was possible in a patient with a left chyle fistula in the neck. After a dose of 50 g liquid paraffin administered as a laxative, 246 ml chyle was collected within the following 14 h which yielded a total of 4.5 mg liquid paraffin. Its composition was identical with the administered laxative. Assuming a daily lymph volume of 1.51, the resorbed amount would correspond to a resorption rate of 0.5 (see article) liquid paraffin. The importance of these results as well as the diagnostic consequences arising from the described analytical technique are discussed in detail.


Purfication of synthetic cardiotoxin by affinity chromatography. A polypeptide containing 60 amino acids with 4 disulphide bonds, synthesized by the solid-phase method, was highly purifed by anticardiotoxin-Sepharose affinity chromatography following gel filtration and CM-cellulose chromatography. The identification of the final product as cardiotoxin was confirmed by thin-layer chromatography on silica gel, polyacrylamide gel electrophoresis, amino acid analysis, circular dichroism spectra, N-terminal analysis and four biological tests.


Gel filtration of phthalate esters.

Gel filtration of phthalate esters. Calibration of the gel-filtration system polystyrene beads-benzene is effected in terms of the molecular weights of solutes without using a void volume indicator. Correction for dead volumes is made by incorporating in the loads a dye producing a peak of accurately known elution volume to serve as reference point on a stripchart absorption diagram of column effluent. The calibrated column separates small amounts of plasticizer phthalate esters from excess of lipids extracted from biological materials, providing evidence of identity in samples sufficiently pure for estimation by gas chromatography. Further purification can, where necessary, be achieved quantitatively by high-performance liquid chromatography on silicic acid with 2% (v/v) diisopropyl ether in n-hexane as mobile phase.


Derivatisation of steroid hormones with t-butyldimethylsilyl imidazole. An efficient procedure for the derivatisation of hydroxyl and alpha,beta-unsaturated ketonic steroids using t-butyldimethylsilyl imidazole is described. The conditions which employ potassium acetate as a basic catalyst result in quantitative formation of BDMS ethers and greater than 96% yield of BDMS-enol ethers. Various mechanisms involving the catalysed and uncatalysed reactions of t-butyldimethylsilyl imidazole with alcohols and enols are also discussed. The mass spectra of the BDMS-enol ethers, in contrast to the BDMS ethers, are dominated by an intense molecular ion, which facilitates the quantitation of these derivatives in biological samples. An example of the method is shown by the determination of 4-androstene-3,17-dione in urine using a stable-isotope internal standard.


Reverse phase high pressure liquid chromatography for the separation of peptide hormone diastereoisomers. Diastereoisomers of specifically labeled oxytocin derivatives were resolved using reverse phase high pressure liquid chromatography. The peptides [1-hemi-DL-[alpha-2H]cystine]oxytocin, [6-hemi-DL-[alpha-2H]cystine]oxytocin, [2-DL-[alpha-2H]tyrosine]oxytocin and[8-DL-[2-13C]leucine]oxytocin were readily separated using the conditions described. The diastereoisomers of the oxytocin analog [3-DL-[2-13C]leucine]oxytocin also demonstrated baseline resolution under the same conditions. The procedure offers the investigator a rapid method for screening synthetic oxytocin peptides for undesirable diastereoisomeric by-products.


Simultaneous determination by GC-MS-SIM of o-, m-, p-hydroxyphenylacetic acid, 3:4-dihydroxyphenylacetic acid and homovanillic acid in biological samples using a common selected ion. A GC-MS-SIM method is described for the simultaneous determination of five acid metabolites, o-, m-, and p-hydroxyphenylacetic acid, 3:4-dihydroxyphenylacetic acid and homovanillic acid in biological samples using a common selected ion for all the acids. The TMS ethers of methyl or ethyl esters of these compounds have a common ion at m/e 179 which is used for their quantitation by selected ion monitoring. The molecular ion at m/e 238 of the three monohydroxy phenylacetic acids, which is also the M-30 ion for homovanillic acid, is also used for their quantitation. The GC conditions for their separation, the relative sensitivities and some advantages of TMS ethers over other derivatives are described. The application of this method to biological samples is illustrated by data on human urine and plasma.


Rapid determination of quinidine in human plasma by high-performance liquid chromatography. A sensitive and specific method for the determination of quinidine in plasma is described. Quinidine is extracted with chloroform, the extract is evaporated and reconstituted with methanol and the methanol extract is analyzed by high-performance liquid chromatography using a silica gel column. The recovery of quinidine from plasma varied between 93.0 to 100%. Quinidine levels as low as 12 ng/ml can be detected by this method.


Vero microcultures for adenovirus neutralization tests. A microculture neutralization test is described for measuring specific antibody levels to the 35 human adenovirus serotypes in Vero cells. The test is read at 5 days by macroscopic observation after staining the uninfected cells with crystal violet. The test is performed with a minimum of manipulations and gives serum titers comparable with those obtained in tube macrocultures of monkey kidney, human embryonic kidney, and Vero cells. The Vero microculture neutralization test measures inhibition of adenovirus toxicity, although selected human adenoviruses serially subpassaged in Vero cells were shown to successfully adapt and replicate in the absence of detectable helper viruses.


New medium for isolation of Actinomyces viscosus and Actinomyces naeslundii from dental plaque. Metronidazole (10 microgram/ml) and cadmium sulfate (20 microgram/ml) were added to a gelatin-based medium to select for microaerophilic Actinomyces species from dental plaque samples. The new medium (GMC), when incubated anaerobically, allowed 98% recovery of seven pure cultures of Actinomyces viscosus and 73% recovery of eight pure cultures of Actinomyces naeslundii, while suppressing 76% of the total count of other organisms in dental plaque samples. In 203 plaque samples, recoveries of A. viscosus and A. naeslundii on GMC and another selective medium for oral Actinomyces (CNAC-20) were compared. Recovery of A. viscosus was comparable on the two media. Recovery of A. naeslundii was significantly higher on GMC than CNAC-20 (P is less than 0.001), and GMC allowed a more characteristic cell morphology of both organisms. GMC medium appears to be useful for the isolation and presumptive identification of A. viscosus and A. naeslundii from dental plaque.


Brain abscess due to Streptococcus MG-intermedius (Streptococcus milleri). Three cases of brain abscesses due to Streptococcus MG-intermedius are reported, and the literature pertaining to this subject is reviewed. The importance of careful and complete identification of these etiological agents of infection is stressed. The clinical presentation, the origin of S. MG-intermedius producing brain abscess, and its relation to hepatic abscesses and endocarditis are discussed.


Endocarditis associated with cardiac catheterization due to a Gram-positive coccus designated Micrococcus mucilaginosus incertae sedis. A gram-positive coccus, presently named Micrococcus mucilaginosus incertae sedis, was isolated from 14 blood cultures from a patient with endocarditis. The first positive blood culture was drawn 5 days after the patient underwent cardiac catheterization.


Lymphocyte responses to hemagglutinin and neuraminidase subunits of influenza virus. The responses of peripheral lymphocytes to purified hemagglutinin and neuraminidase subunits and other components of an influenza. A virus were measured in 21 normal adults and compared with antibody titers. All had influenza antibodies and demonstrated influenza antigen recognition by lympho-proliferative responses. There was a significantly greater response to the two purified influenza virus surface antigens, hemagglutinin and neuraminidase, than to either whole intact virus or its separated subviral core. No correlation between magnitudes of antibody titers and lymphocyte responsiveness was observed.


Atypical Yersinia enterocolitica: clinical and epidemiological parameters. Infections due to biochemically typical Yersinia enterocolitica usually present as gastroenteritis, mesenteric lymphadenitis, terminal ileitis, and septicemia often with visceral abscesses. In these instances, the isolates have been biochemically typical and of well-established serotypes, namely 0:3 or 0:9 and, in the United States, 0:5 or 0:8. The recovery, recognition, and significance of biochemically and serologically atypical Y. enterocolitica in human infections has proceeded more slowly. From an analysis of the clinical histories of 20 patients infected with 21 such aberrant Y. enterocolitica, it appears that these strains are of restricted pathogenic potential, producing various clinical entities such as localized skin abscesses, conjunctivitis, self-limiting enteritis, and wound and urinary tract infections in hosts with predisposing factors. Epidemiologically, whereas episodic acquisition of atypical strains by hospitalized patients is indicative of nosocomial transmission, in the present series sporadic isolations over a 4-year period, mainly from ambulatory patients, suggest an occult reservoir in the community serviced by The Mount Sinai Hospital. In contrast to typical Y. enterocolitica, which has become well adapted in animal and human hosts, it appears that environmental strains may be in the evolutionary process of becoming adapted to humans.


Disseminated infection by Fusarium moniliforme during treatment for malignant lymphoma. Disseminated infection caused by Fusarium moniliforme is described in a 32-year-old granulocytopenic man with malignant lymphoma being treated with cytotoxic drugs and corticosteroids. Infected skin denuded by antecedent severe varicella-zoster infection was the probable source of fungemia. F. moniliforme grows rapidly on common mycological media as a lavender- to violet-colored mold at 25 to 37 degrees C. Its aerial hyphae produce fusoid macroconidia and characteristic fusiform microconidia in chains. The morphology of hyphae in tissue closely resembles species of Aspergillus and is not diagnostically specific. Morphological characteristics which distinguish cultures of F. moniliforme from other medically important species of Fusarium are discussed.


Diagnosis of California La Crosse virus infection by counterimmunoelectrophoresis. La Crosse virus cell culture-derived soluble antigen was used in a counterimmunoelectrophoresis procedure to assess the presence or absence of La Crosse antibodies in 79 paired acute- and convalescent-phase human sera. The counterimmunoelectrophoresis test appeared to measure the same antibody as the complement fixation test but was more sensitive and rapid.


Oligomeric immunoglobulin A antibody response to rubella virus infection. Pooled sera from rubella patients in the early convalescent stage, containing a high titer of hemagglutination-inhibiting (HI) antibody, were treated with protein A-conjugated gel to reduce immunoglobulin G (IgG) antibody and then centrifuged in sucrose gradients. This treatment resulted in the detection of an HI activity peak sedimenting at a rate intermediate between 7S and 19S. In contrast to the 19S antibody, the HI activity of this peak was not abolished by 2-mercaptoethanol, but sedimented at 7S after this treatment. The activity was considered to consist of IgA oligomers, since it was removed by anti-IgA immunosorbent. The appearance of the oligomeric IgA antibody after the infection was then studied using serum samples collected sequentially from five rubella patients. Shortly after the onset of the disease, the HI activity appeared at high titer and thereafter gradually decreased in titer until it could no longer be detected in the sera. The time of its disappearance varied with each patient.


Group IVe-like gram-negative bacillemia in a patient with obstructive uropathy. A nonfermentative, gram-negative bacillus, resembling Center for Disease Control group IVe, was isolated from the blood of a patient with obstructive uropathy. Prior studies implicating this organism in septicemia could not be found in the literature. It is suggested that this is the first time that this bacillus has been reported to cause bacillemia in humans.


Usefulness of serial antibody determinations in diagnosis of candidiasis as measured by discontinuous counterimmunoelectrophoresis using HS antigen. We have followed the candida antibody response in 115 patients with different types of candidiasis by discontinuous counterimmunoelectrophoresis using HS antigen to learn whether any early antibody response occurs in systemic candidiasis and whether there are differences in the antibody response in candidiasis involving different organ systems. We found that 23 of 32 (72%) patients with systemic candidiasis had a rise in antibody titer within the first 2 weeks of infection and that high titers were relatively insensitive indicators of infection. No differences were seen in the antibody response in different types of candidiasis. Patients with aspergillosis and torulopsosis had titer rises as well which were attributed to either inapparent candida infection or cross-reacting antibody. A rise in titer was not seen in any patient with candida colonization, bacterial or viral infection, or no infection in contrast to titers greater than or equal to 1/1 or greater than or equal to 1/8 which were seen in these conditions.


Improved media for growth and aerotolerance of Campylobacter fetus. The microaerophilic nature of Campylobacter fetus has complicated its recovery from human and animal sources. In this study, modifications of brucella agar and broth were tested for enhancement of growth and aerotolerance of 64 strains of C. fetus, representing each subspecies. Brucella agar supplemented with 0.025% each FeSO4 7H2O, sodium metabisulfite, and sodium pyruvate, supported growth of 98, 77, and 63% of the strains at 6% O2, 17% O2, and 21% O2, respectively. Unsupplemented brucella agar supported growth of 94, 48, and 20% of the strains. Brucella broth supplemented with 0.2% FeSO4.7H2O, 0.025% sodium metabisulfite, and 0.05% sodium pyruvate supported growth of 98% of the strains at 21% O2, compared to 75% with unsupplemented brucella broth. With both the supplemented agar and broth, growth responses occurred 1 to 2 days earlier than usual. Growth and aerotolerance of three strains of Campylobacter sputorum subsp. bubulus were not enhanced by the supplements.


Identification of Achromobacter species by cellular fatty acids and by production of keto acids. The cellular fatty acid composition and metabolic products of 12 reference strains of Achromobacter sp. and A. xylosoxidans were determined by gas-liquid chromatography (GLC). Results showed that the two Achromobacter groups are strikingly different and can be readily distinguished on the basis of cellular fatty acids and the short-chain acids produced by Achromobacter sp. The major cellular fatty acids of Achromobacter sp. were octadecenoic (18:1) and a 19-carbon cyclopropanoic (19:0 delta) acid, whereas hexadecanoic (16:0) and a 17-carbon cyclopropanoic (17:0 delta) acid were principal components of the lipids of A. xylosoxidans. Hydroxy acids were not found in strains of Achromobacter sp. but comprised approximately 20% of the cellular fatty acids of A. xylosoxidans. In addition, Achromobacter sp. produced relatively large amounts of 2-ketoisocaproic acid, which was detected in only trace amounts from strains of A. xylosoxidans. The data show that GLC tests provide additional criteria for differentiating groups which are very closely related when evaluated with conventional tests. The GLC tests can be readily adapted in the clinical laboratory because they are rapid, highly reproducible, relatively inexpensive, and simple to perform.


Yersinia enterocolitica: serotypes and biotypes isolated from humans and the environment in Quebec, Canada. Thirty-one strains of Yersinia enterocolitica were isolated from food and surface water. During the period of January 1975 to June 1977, 157 strains from 143 human cases were also isolated. Among the different serotypes from nonhuman sources, serotypes 6,30 and 4,32 were the most common. Serotype 4,32 was present only in food and not in water. Serotype 3 was isolated only from humans. Among the different serotypes from human cases, 73.9% belonged to serotype 3. Only serotype 3 was isolated from children under 4 years old. The presence of other serotypes increased and that of serotype 3 decreased in frequency as the age progressed. No serotype 3 was isolated from human cases aged 50 years and more.


Occurrence of mycoplasmas in urinary tracts of patients with acute pyelonephritis. Mycoplasma hominis was isolated from the upper urinary tract in 7 of 80 patients with acute pyelonephritis and from 0 of 60 patients with noninfectious diseases of the urinary tract, a significant difference. In four cases M. hominis was isolated in pure culture, in one it was isolated together with Ureaplasma urealyticum, and in two it was isolated with bacteria. U. urealyticum was isolated from the upper urinary tract of five patients, all with acute pyelonephritis; this was not significantly different from the control group.


Cerebrospinal fluid lactic acid in diagnosis of meningitis. Quantitative lactate determinations were performed on cerebrospinal fluids to assess their value in the rapid diagnosis of bacterial and mycotic meningitis and to evaluate their value in assessing the prognosis in these patients. Cerebrospinal fluid lactate concentrations were elevated in all patients with untreated bacterial or fungal meningitis. Lactate concentrations proved very valuable in following patients with mycotic meningitis and in differentiating aseptic from bacterial meningitis. Elevated cerebrospinal fluid lactate is not specific for meningitis. Lactate is also elevated in situations where there is central nervous system ischemia and necrosis and in patients with brain tumors. Lactate concentration is normal in chronic degenerative brain diseases. Thus, the clinical situation must be taken into account when interpreting the lactate concentrations.


Survival of 125iodine-labeled Factor VIII in normals and patients with classic hemophilia. Observations on the heterogeneity of human Factor VIII. Radiolabeled human Factor VIII was used to study its survival in normals and patients with classic hemophilia, and to study the heterogeneity of Factor VIII; Purified Factor VIII was radiolabeled with 125iodine (125I-VIII) without loss of its structural integrity. The survival of 125I-VIII was studied in six normals and six hemophiliacs of whom four of the hemophiliacs had received transfusions with normal cryoprecipitate before the 125I-VIII infusion. No significant difference was observed between the disappearance of Factor VIII coagulant activity and radioactivity in these hemophiliacs. 125I-VIII in plasma showed a biphasic disappearance with an average t1/2 of 2.9 +/- 0.4 h (SEM) for the first phase and 18.6 +/- 0.7 h (SEM) for the second phase, respectively. The survival of 125I-VIII was similar comparing normals and hemophiliacs. The highest molecular weight forms of Factor VIII disappear more rapidly than the lower molecular weight ones. This was established by analysis of the fractions obtained by gel chromatography of plasma collected at several times after infusion and by analysis of the in vivo disappearance of three subfractions of Factor VIII. The fraction of 125I-VIII binding to platelets in the presence of ristocetin (containing the highest molecular weight forms of Factor VIII including the ristocetin cofactor) represented about 50% of the radioactivity present in plasma after infusion and showed a t 1/2 of 11.7 +/- 0.9 h (SEM) for the second phase. The fraction, which was recovered in cryoprecipitate of the recipient's plasma, represented about 90% of the initial radioactivity and showed a t 1/2 of 16.3 +/- 0.8 h (SEM) for the second phase. The fraction of 125I-VIII remaining in the cryosupernatant plasma (containing low molecular weight forms of Factor (VIII) showed a t 1/2 of 27.2 +/- 1.1 h (SEM). The first phase of the disappearance of 125I-VIII is caused in part by the disappearance of the highest molecular weight forms, which are possibly removed by the reticuloendothelial system.


A common uptake system for serotonin and dopamine in human platelets. Kinetic and pharmacologic properties of uptake of serotonin and dopamine by normal human platelets have been investigated to test whether platelets can be employed as a model system for the reuptake of serotonin and dopamine in brain. Uptake of serotonin into platelets closely resembles reuptake of serotonin into serotonergic neurons. In contrast, uptake of dopamine into platelets appears to be mediated inefficiently via the specific serotonin uptake mechanism, based upon several lines of evidence. Serotonin and dopamine compete with each other, Antidepressant drugs, which are competitive inhibitors of uptake of both of these neurotransmitters, act at the same concentration of drug despite large differences in the Km values. Serotonin antagonists inhibit both serotonin and dopamine uptake. Finally, a serotonin-specific uptake inhibitor (fluoxetine) blocks dopamine, as well as serotonin, uptake.


Gentamicin blockade of slow Ca++ channels in atrial myocardium of guinea pigs. Cardiac dysfunction is occasionally detected in patients undergoing treatment with amino-glycoside antibiotics, however, the mechanism responsible for the negative inotropic effect of these agents has not been identified. In the present investigation electrically driven left atria of guinea pigs were used to study the effects of gentamicin on calcium ion (Ca++)-dependent contractile events in heart muscle isolated from in vivo influences. When atria were first inactivated by excess potassium ion (K+; 22mM) and contractions were then restored by isoproterenol (an experimental model that accentuates the contractile dependence of myocardial fibers on influx of Ca++ through specific "slow channels" of the sarcolemma), the cardiac depressant activity of gentamicin (0.1 mM) was profoundly augmented. Conversely, the negative inotropic effect of tetrodotoxin (23.5 micron) was abolished by the same experimental conditions. Also, gentamicin (1 mM) and La+++ (0.5 mM) markedly decreased the positive inotropic response to increased frequency of stimulation; whereas, D600 (1.05 micron) converted the positive frequency-force relationship to a negative relationship. Present data indicate a direct cardiac depressant action of gentamicin, and suggest that this antibiotic adversely affects either the transport system responsible for Ca++ movement through slow channels of the sarcolemma, the availability of Ca++ for translocation to these sites, or both.


Maturation of jejunum and ileum in rats. Water and electrolyte transport during in vivo perfusion of hypertonic solutions. During osmotic diarrhea, loss of water and electrolytes appears to be greater in infants than in adults. In 2-, 3-, and 7-wk-old rats, we studied net transport of H(2)O, Na, and Cl, during in vivo perfusion of segments of the jejunum and ileum, from solutions with osmolalities of 300, 375, 500, or 700 mosmol/kg. In the jejunal segments, from the hypertonic solutions net transport of H(2)O, Na, and Cl was into the lumen and greater in the 2- than 7-wk-old rats. In the ileal segments, transport of water was into the lumen, transport of Na was minimal and variable, whereas transport of Cl was usually out the lumen. In 3-wk-old rats, transport rates were intermediate between those in 2- and 7-wk-old rats. The calculated filtration coefficient (microliters of H(2)O transported per hour per unit osmolality gradient-lumen-serum-per gram dry weight) of water suggested that the resistance to water flow did not increase with rise in luminal hypertonicity in the jejunum of the 2- and 3-wk-old rats, whereas in jejunum of the 7-wk-old rats and in ileum of rats in all three ages, the resistance to water flow increased with the rise in luminal osmolality. The differences in the transport rates and the resistance to water flow, between segments of the 2-, 3-, and 7-wk-old rats, suggested a maturational phenomenon that appears to continue beyond the 3rd wk of life and could have been due to differences in some physical property of the mucosal membrane.


Further studies on segmental sodium transport in the rat kidney during expansion of the extracellular fluid volume. The present studies were designed to further investigate the possibility of heterogeneity of nephron function during Ringer loading in the rat, and to determine the specific nephron segment responsible for this finding. As in previous studies from this laboratory with smaller rats (50-125 g), net addition of sodium between late distal tubule and papillary base (6.9 vs. 10.4% of the filtered load, respectively, P <0.005) was found in more mature rats (170-230 g). In contrast, there was net reabsorption of sodium between these two segments in nonvolume-expanded animals, 1.70 vs. 0.45% of the filtered sodium load, P <0.005. Because nephron heterogeneity of sodium transport during extracellular volume expansion is the most likely explanation for these findings, further studies were performed to determine the specific juxtamedullary nephron segment responsible for the net addition pattern between late distal tubule and papillary base in Ringer-loaded animals. First, a comparison was made of sodium delivery to the late proximal tubule of superficial nephrons vs. the delivery rate to the bend of Henle's loop of juxtamedullary nephrons in both hydropenia and Ringer loading. Fractional sodium delivery was quite comparable between the superficial and juxtamedullary nephrons in both hydropenia and Ringer loading although the absolute level was much greater in both groups of nephrons in the Ringer studies. Chlorothiazide (15 mg/kg loading and 15 mg/kg per h) given during Ringer loading markedly increased late distal sodium delivery, 19% of the filtered load, but did not prevent net addition of sodium at the papillary base. In contrast, furosemide (5 mg/kg loading and 5/mg/kg per h) given during Ringer loading completely reversed the segmental pattern, 35.5 and 28.8% at late distal tubule and papillary base, respectively, P <0.005. These studies demonstrate that the net addition of sodium between late distal tubule and papillary base during Ringer loading is not limited to immature rats and that the segmental pattern does not occur in non-volume-expanded animals. Further, the reversal of the net addition pattern with furosemide, but not chlorothiazide, and the comparable proximal nephron delivery rates in Ringer loading suggest that the loop of Henle of juxtamedullary nephrons reabsorbs less sodium than the same portion of superficial nephrons in this setting. A model is proposed to explain this finding.


Direct determination of PCO2 in the rat renal cortex. The mechanism by which the kidney reabsorbs sodium bicarbonate could be a result of (a) H+ secretion, (b) direct HCO3- reabsorption, or (c) a combination of both processes. Most of the studies which have supported the H+ secretory theory have involved the assumption that tubular fluid and arterial PCO2 were equal. We have utilized a new PCO2 microelectrode to directly determine in situ PCO2 of tubular fluid and stellate vessel blood in the cortex of the rat kidney during control conditions and after alterations in acid-base status. In 21 control rats, proximal tubular fluid PCO2 exceeded systemic arterial PCO2 (deltaCO2) by 25.9 +/- 0.92 mm Hg (P less than 0.001). The values obtained for both distal tubular fluid and stellate vessel blood were not significantly different from proximal tubular PCO2. Evaluation of PCO2 in the proximal tubules of Munich-Wistar rats did not reveal evidence for a declining profile for PCO2 along the length of the nephron. When proximal bicarbonate reabsorption was increased or decreased acutely by alterations in acid-base status, deltaPCO2 changed in paralle. Furthermore, benzolamide administration significantly reduced deltaPCO2. We conclude: (a) that the PCO2 in tubular fluid is significantly greater than systemic arterial PCO2, (b) that there is no tendency for the observed PCO2 to fall along the proximal tubule, (c) the mean PCO2 in the proximal and distal tubules as well as the stellate vessle is not significantly different, thereby rendering the concept of a "diffusion barrier" for CO2 in the proximal tubule unlikely, and (d) the level of renal cortical PCO2 appears to vary directly with the magnitude of bicarbonate reabsorption.


Requirements for the solubilization of immune aggregates by complement. The role of the classical pathway. In this paper we examine the role of the classical pathway in the complement-mediated solubilization of immune precipitates (CRA). Serum reagents were depleted of the alternative pathway components properdin and factor D. Both depleted reagents lack CRA although they have almost intact hemolytic activity. Also, immune complexes were not solubilized when incubated with high concentrations of the classical pathway components (C1, C4, C2, and C3. We conclude that CRA is not mediated by the classical pathway alone. Activation of the classical pathway by the immune aggregates greatly enhances CRA. The effect of the classical pathway is to deposit C3b on the antigen-antibody lattice and promote the assembly of a lattice-associated, properdin-dependent C3-convertase. Although C3, C4, and properdin were detected on complexes solubilized by serum in the presence of Ca++ and Mg++, only C3 and properdin were found on the complexes when Ca++ had been chelated by ethylene glycol-bis-(beta-aminoethyl ether), N,N'-tetraacetic acid. In both situations the aggregates were capable of converting C5 in the fluid phase. However, no C5 was found on the solubilized complexes. These findings suggest that in contrast to nascent C3b and C4b, nascent C5-9 lacks binding affinity for immune aggregates.


Multiple forms of human plasma renin substrate. The objective of this investigation was to determine whether heterogeneity of plasma renin substrate could be observed in states of steroid excess and various forms of hypertensive disease. In states of stimulated renin substrate production by estrogens or glucocorticoids, multiple forms of renin substrate were apparent when stimulation was excessive. Stimulation of substrate production caused by uremia associated with hypertension showed similar results. None, or only trace quantities of the additional forms of renin substrate were evident in subjects with normal or suppressed levels of plasma renin substrate. The additional forms of renin substrate could be distinguished from the normal form on the basis of cross-reactivity with a specific antiserum to the normal form, electrophoretic mobility, and kinetic rate constants. Differences in rate constants of the various forms of plasma renin substrate may account for the altered rate of the renin reaction associated with several states of hypertension. In plasma of patients with renovascular hypertension, significant quantities of a protein which cross-reacted with the antiserum but could not generate angiotensin I were observed.


Sodium, phosphate, glucose, bicarbonate, and alanine interactions in the isolated proximal convoluted tubule of the rabbit kidney. Interactions among the transport systems involved with sodium, bicarbonate, glucose, phosphate, and alanine absorption in isolated segments of the rabbit proximal convoluted tubule were examined with radioisotopic techniques to measure glucose, phosphate, and fluid absorption rates. The composition of the perfusate and bath varied from normal, physiological fluids to fluids deficient in a single solute. The deletion of glucose from the perfusate increased the lumen-to-bath flux of phosphate from 5.51 +/- 1.15 to 8.32 +/- 1.34 pmol/mm-min (P less than 0.01). Similar changes occurred when glucose transport was inhibited by phlorizin 10 micron in the perfusate, The deletion of alanine from the perfusate increased the lumen-to-bath flux of phosphate from 6.55 +/- 1.08 to 9.00 +/- 1.30 pmol/mm-min (P less than 0.01) but did not affect glucose transport significantly, 80.1 +/- 10.1 vs. 72.5 +/- 5.4 pmol/mm-min. Replacement of intraluminal sodium with choline, elimination of potassium from the bath, and removal of bicarbonate from the lumen and bath each reduced glucose, phosphate, and fluid absorption. These data indicate that the proximal absorptive processes for glucose and for phosphate include elements that are dependent upon some function of sodium transport. Additionally, the effects on phosphate transport of deleting glucose or alanine occur independent of any changes in net sodium transport and are opposite the effects of deleting bicarbonate. These differences may relate to the observations that the transport of glucose and alanine is electrogenic while that of bicarbonate is not. Regardless of possible mechanisms, the data demonstrate that important changes in the absorption rates of different solutes handled significantly by the proximal convoluted tubule may occur in response to changes in specific components of proximal sodium transport.


Glucose and alanine metabolism in children with maple syrup urine disease. In vitro studies have suggested that catabolism of branched chain amino acids is linked with alanine and glutamine formed in, and released from, muscle. To explore this possibility in vivo, static and kinetic studies were performed in three patients with classical, and one patient with partial, branched chain alpha-ketoacid decarboxylase deficiency (maple syrup urine disease, MSUD) and compared to similar studies in eight age-matched controls. The subjects underwent a 24-30-h fast, and a glucose-alanine flux study using stable isotopes. Basal plasma leucine concentrations were elevated (P <0.001) in patients with MSUD (1,140+/-125 muM vs. 155+/-18 muM in controls); and in contrast to the controls, branched chain amino acid concentrations in plasma increased during the fast in the MSUD patients. Basal plasma alanine concentrations were lower (P <0.01) in patients with classical MSUD (153+/-8 muM vs. 495+/-27 muM in controls). This discrepancy was maintained throughout the fast despite a decrease in alanine concentrations in both groups. Plasma alanine and leucine concentrations in the patient with partial MSUD were intermediate between those of the controls and the subjects with the classical form of the disease. Circulating ketone bodies and glucoregulatory hormones concentrations were similar in the MSUD and normal subjects during the fast. Alanine flux rates in two patients with classical MSUD (3.76 and 4.00 mumol/Kg per min) and the patient with partial MSUD (5.76 mumol/Kg per min) were clearly lower than those of the controls (11.72+/-2.53 [SD] mumol/Kg per min). After short-term starvation, glucose flux and fasting concentrations were similar in the MSUD patients and normal subjects.These data indicate that branched chain amino acid catabolism is an important rate limiting event for alanine production in vivo.


The influence of fasting, diabetes, and several pharmacological agents on the pathways of thyroxine metabolism in rat liver. As judged from both paper and column chromatography, slices or homogenates of liver from rats fasted for 48 h displayed a lesser rate of generation of (125)I-labeled 3,5,3'-triiodothyronine (T(3)) from (125)I-labeled thyroxine (T(4)) added to incubation media than did preparations from normal chow-fed animals. A similar defect in the conversion of T(4) to T(3) in the livers of fasted animals was observed when preparations were incubated with substrate concentrations of T(4) so that T(3) generation could be assessed by radioimmunoassay. The effect of fasting could be prevented, wholly or in part, by administration of glucose in the drinking water to otherwise fasted animals, and the degree of prevention appeared to be proportional to the concentration of glucose employed. Diminished generation of T(3) from T(4) was similarly evident in the livers of animals with streptozotocin-induced diabetes mellitus, and this defect was overcome by the provision of insulin in vivo, but not in vitro. Decreased formation of T(3) from T(4) was also observed in preparations of liver from animals given dexamethasone, amiodarone, and propylthiouracil. In no case could these effects on the net formation of T(3) from T(4) be explained by effects of the experimental conditions on the degradation of the T(3) generated, as judged from the rate of degradation of exogenous (125)I-T(3) measured in parallel incubates. An analysis of the rate of disappearance of (125)I-T(4) from reaction mixtures in relation to the rate of appearance of (125)I-T(3) and (125)I-iodide was employed to estimate the activity of the 5-monodeiodinating pathway of T(4) metabolism that leads to the formation of 3,3',5'-triiodothyronine (reverse T(3)). Such estimates indicated that reverse T(3) formation was actively proceeding in the preparations studied, was slightly enhanced by fasting, was unaffected by dexamethasone and amiodarone, and was markedly inhibited by propylthiouracil. In view of the similarities between the effect of these experimental manipulations on the generation of T(3) from T(4) by rat liver in vitro to their effects on the production rates and serum concentrations of T(3) in man, it is concluded that the rat liver system provides a suitable model for the study of factors that influence the conversion of T(4) to T(3) in man. In addition, the findings strongly indicate that this process, at least in the liver, is closely linked to the utilization of carbohydrate.


Responsiveness of neoplastic and hyperplastic parathyroid tissues to calcium in vitro. Secretory and biosynthetic responses of adenomatous, carcinomatous, and hyperplastic parathyroid tissues to variable concentrations of extracellular calcium were assessed in vitro. Tissues, obtained at the time of parathyroidectomy, were incubated for 4 h in media containing radioactive amino acids and varying (0.5-5.0 mM) concentrations of calcium. Amounts of newly synthesized and total parathyroid hormone and proparathyroid hormone in extracts of tissues and media were measured by polyacrylamide gel electrophoresis and by radioimmunoassay, respectively. All tissues studied (six adenomas, two specimens of chief-cell hyperplasia, one carcinoma, and normal bovine and human glands) responded to changes in calcium concentrations; decreasing concentrations of calcium stimulated release and decreased tissue storage of hormone. Six of the abnormal tissues required greater than normal concentrations of calcium (1.8-2.4 mM for 50% of effect) to elicit secretory responses comparable with those of normal glands (1.4 mM). Maximum effects of calcium on release of hormone varied from 2- to 10-fold among different tissues. Release of some hormone persisted even in concentrations of calcium  as high as 5.0 mM. Relative amounts of hormone released from and retained in the tissues varied greatly among the tissues, as did the absolute amounts of hormone produced; newly synthesized, labeled hormone ranged between 0.6 and 12% of total labeled protein, and immunoreactive hormone ranged between 0.015 and 0.9% of total tissue protein. Effects of calcium on hormone biosynthesis, as determined by analyses of amounts of proparathyroid hormone in the tissues, were variable among tissues and in many cases were negligible. These results indicate that neoplastic and hyperplastic parathyroid tissues retain secretory responsiveness to changes in extracellular concentrations of calcium. Responses, however, are highly variable among different tissues, and in many instances are abnormal, inasmuch as greater than normal concentrations of calcium are required to alter release and synthesis of hormone. A combination of both increased mass of glandular tissue and abnormal regulations of hormone secretion appear to contribute to the hypersecretion of hyperparathyroidism.


The metabolic response to hypocaloric protein diets in obese man. Exogenous protein in the absence of other calories can cause protein-sparing, but the mechanisms involved are controversial. It has been postulated that low insulin and high fat-derived substrate levels are necessary and sufficient conditions for such protein-sparing. We therefore established such conditions with differing protocols of protein input to define the role of protein input in mediating the response. Three groups of obese, nondiabetic subjects received the following diets: (1) 82.5+/-1.0 g protein/day (400 cal/day) for 21 days, n = 7; (2) the same, but as a refeeding diet for 7 days after 21-28 days of total fasts, n = 7; and (3) commencing with the same input, but with daily stepwise decrements over 14 days to 19.4+/-2.2 g/day, then maintained an additional 7 days, n = 4. Diet 3 gave approximately the amount and pattern of protein lost during total fasting. The circulating hormone and substrate responses of diets 1 and 3 were comparable and resembled those of total fasts, in that plasma glucose and insulin fell and free fatty acids rose. Blood levels of alanine, pyruvate, and other glucogenic amino acids fell and blood levels of branched-chain amino acids rose transiently. Blood 3-hydroxybutyrate levels and urinary excretion were greater in diet 3 than diet 1, but less than in total fasting. Nitrogen balance in diet 1 was transiently negative, but in equilibrium from 12 to 21 days. In diet 3, it was constantly negative at -6 g/day, the values also observed at 21 days of fasting. Mean 3-methylhistidine excretion decreased by 170 mumol/day in diet 1 and 107 mumol/day in diet 3, reflecting decreased muscle protein catabolism. The refed, protein-depleted subjects, diet 2, showed an increase in plasma glucose without alteration in insulin levels. Free fatty acid and ketone body levels decreased to those of the steady state observed in diet 1. Glucogenic and branched-chain amino acids decreased transiently. Nitrogen balance became positive, and the low 3-methylhistidine excretion increased by 152 mumol/day. The differing responses of nitrogen balance could not be accounted for on the basis of levels of insulin or of fat-derived substrates. The primary determinants of the protein-sparing observed appeared to be the protein supply itself, and the magnitude of the decrease in endogenous protein catabolism. The positive balance on refeeding after prior depletion of protein stores was likely due to the exogenous supply, combined with decreased catabolism and considerably increased reutilization.


Glucose disposal during insulinopenia in somatostatin-treated dogs. The roles of glucose and glucagon. The first aim of this study was to determine whether the plasma glucose level can regulate hepatic glucose balance in vivo independent of its effects on insulin and glucagon secretion. To accomplish this, glucose was infused into conscious dogs whose basal insulin and glucagon secretion had been replaced by exogenous intraportal insulin and glucagon infusion after somatostatin inhibition of endogenous pancreatic hormone release. The acute induction of hyperglycemia (mean increment of 121 mg/dl) in the presence of basal levels of insulin (7+/-1 muU/ml) and glucagon (76+/-3 pg/ml) resulted in a 56% decrease in net hepatic glucose production but did not cause net hepatic glucose uptake. The second aim of the study was to determine whether a decrease in the plasma glucagon level would modify the effect of glucose on the liver. The above protocol was repeated with the exception that glucagon was withdrawn (83% decrease in plasma glucagon) coincident with the induction of hyperglycemia. Under this circumstance, with the insulin level basal (7+/-1 muU/ml) and the glucagon levels reduced (16+/-2 pg/ml), hyperglycemia (mean increment of 130 mg/dl) promoted marked net hepatic glucose uptake (1.5+/-0.2 mg/kg per min) and glycogen deposition. In conclusion, (a) physiological increments in the plasma glucose concentration, independent of their effects on insulin and glucagon secretion, can significantly reduce net hepatic glucose production in vivo but at levels as high as 230 mg/dl cannot induce net hepatic glucose storage and (b) in the presence of basal insulin the ability of hyperglycemia to stimulate net hepatic glucose storage is influenced by the plasma glucagon concentration.


Analysis of linkage between the major histocompatibility system and juvenile, insulin-dependent diabetes in multiplex families. Reanalysis of data. Linkage analysis between the major histocompatibility system (HLA) and juvenile, insulin-dependent diabetes, assuming an autosomal recessive mode and 50% penetrance was performed on 21 juvenile, insulin-dependent diabetic multiplex families (two or more diabetics per sibship) with phenotypically normal parents. The total lod score was the highest (3.98) at a recombination fraction of 13%. For a penetrance of 100%, the highest total lod score was 2.92 at a recombination fraction of 18%. These results are compatible with the existence of linkage between an autosomal recessive diabetic gene with 50% penetrance and the HLA in some of the families studied. Our ascertainment strategy would be expected to increase the likelihood of selecting for genetically homogenous diabetes and against sporadic forms of the disease. Thus, our findings may apply only to a small proportion of all cases of juvenile, insulin-dependent diabetes.


Measurement of total haemolytic complement activity in body fluids. The measurement of total haemolytic complement is of diagnostic value in the rheumatic diseases. A method which is easier to perform than that of Kabat and Mayer (Experimental Immunochemistry, 2nd ed., Thomas, Springfield, Ill.) is described. A unit based on the mean value of the complement activities of normal sera is proposed to replace CH50 units. Other steps by which standardisation of the assay may be improved are suggested.


Determination of antibiotic sensitivities by the Sensititre system. The Sensititre system provides antibiotics dried in microtitre trays in appropriate concentrations for the determination of minimum inhibitory concentrations (MIC) when organisms are added in liquid culture medium and incubated overnight. Results with a variety of media--for Gram-negative bacilli tested with ampicillin, carbenicillin, cephaloridine, and gentamicin, staphylococci with penicillin, cloxacillin, fusidic acid, and gentamicin, and streptococci with penicillin, ampicillin, cephaloridine, and erythromycin--were usually very similar to those of agar dilution tests. Discrepant results are found in a few situations, and in some cases, especially those involving borderline resistance to fusidic acid and gentamicin among staphylococci and Gram-negative bacilli, we have been unable to suggest modfication to produce the right results. Despite these problems, fortunately few and identifiable, we recommend the system especially for use in laboratories that determine the MICs infrequently.


An assessment of the Sensititre method for determining minimum inhibitory and bactericidal concentrations. Minimum inhibitory and bactericidal concentrations of various antibiotics were determined for 90 strains of five organisms by Sensititre and compared with those obtained by conventional methods. Results by both methods corrleated well. The advantages and limitations of Sensititre are discussed.


Phagocytosis and killing of staphylococci by human polymorphonuclear and mononuclear leucocytes. The phagocytosis and killing of 3H-thymidine-labelled Staphylococcus aureus by polymorphonuclear leucocytes (PMNs) and monocytes (MNs) obtained from 50 health donors were evaluated. In addition, extracellular factors that might influence phagocytosis and killing were studied. The method described gave highly reproducible results. No significant difference was observed in the phagocytic and killing functions of a single donor's PMNs and MNs when studied several times in one day and longitudinally over a period of 1-12 weeks for six donors tested. Likewise, no signigicant difference in uptake and killing was observed when bacteria were opsonised with sera from 11 different normal donors. When Staph. aureus opsonised with normal serum was added to the leucocytes in a ratio of 10 bacteria: 1 leucocyte, the uptake by PMNs and MNs from 50 donors after 20 minutes' incubation was 85% +/- 7 standard deviation (SD) (range 75-98%) and 69% +/- 11 SD (range 54-90%), respectively. The rate of uptake by MNs in the first three minutes of the assay period was only 60% of that by PMNs.


Serum immunoglobulin concentrations before and after splenectomy in patients with homozygous beta-thalassaemia. Serum IgM, IgA, and IgG concentrations were measured in 19 patients suffering from blood-dependent homozygous beta-thalassaemia. The study was prospective and the same patients served as presplenectomy controls. IgG levels rose rapidly and continuously after splenectomy IgA levels also rose but not so sharply. IgM fell to low levelr pattern was found in the cross-sectional part of the study comprising 39 splenectomised and 51 non-splenectomised patients who were tested once.


Cytological basis of histological typing of diffuse Hodgkin's disease. Demonstration of an implied misnomer in the terminology of the Rye classification. Differential cell counts were made on nine lymph nodes whose structure was replaced by diffuse Hodgkin's disease; two of these nodes had the classical histological appearance of the lymphocytic predominance subtype, four of the mixed cellularity subtype, and three of the lymphocytic depletion subtype. Our attempts to achieve valid sampling methods are recorded. The counts, in general, confirm the postulated histological basis of the Rye classification of the subtypes of the diffuse disease. The major discrepancy is that, contrary to the histological descriptions, our direct counts have shown that lymphocytes, are, in general, more numerous in the lymphocytic depletion than in the mixed cellularity subtypes. The cell counts also show that normal mononuclear cells (mainly fibroblasts and macrophage-type cells) are much more numerous in the mixed cellularity subtype than in the other forms of diffuse Hodgkin's disease; this feature has not been emphasised in the Rye classification. On the basis of our differential counts, a hypothesis is proposed that could explain the natural history of the different subtypes of diffuse Hodgkin's disease as the resultant of three processes: (a) tumour aggressiveness, (b) specific cell-mediated immunological reactions, and (c) non-immunological stromal responses.


A case of Lennert's lymphoma.

A case of Lennert's lymphoma. A 73-year-old man who presented with a tonsillar mass and generalised lymphadenopathy died two months after admission to hospital. The appearances in the surgical biopsy material resembled those of Lennert's lymphoma, while the necropsy findings were those of a diffuse lymphocytic lymphoma composed mainly of cells with small nuclei showing only slight irregularity. The implication of these findings in relation to the nature of Lennert's lymphoma are briefly discussed.


Overlap in the spectrum of non-specific inflammatory bowel disease--'colitis indeterminate'. It is stated that 10-20% of cases of non-specific inflammatory bowel disease cannot be classified. Thirty such cases, designated colitis indeterminate at the time of colectomy, were identified from the pathology files of St. Mark's Hospital. The Histopathological features of the surgical specimens and any available biopsy specimens were studied. In nearly all the cases urgent surgery had been required and the features of incipient or established fulminating disease were present. The pathology of these cases of Crohn's disease and ulcerative colitis overlapped, and differentiating features were scant or unreliable. Accepted criteria of Crohn's disease--namely, fissuring ulceration, transmural inflammation, and a maintained goblet-cell population--were found in cases subsequently proved to be ulcerative colitis. Disease activity greatly affected the evaluation of morphological features. Many of the difficulties were resolved when biopsy material obtained during a quiescent phase was examined. The specimens gave a dynamic perspective of the disease process, often more valuable than the static, non-specific picture of acute disease seen in the surgical specimens. Case of colitis indeterminate form a small distinctive group in the spectrum of inflammatory bowel disease which is characterised by a common pattern of pathology that presents a diagnostic dilemma.


Use of a mucolytic agent (Cytoclair) in the preparation of cell material for the detection of malignant cells in sputum. A mucolytic agent, methyl cysteine HC1 (Cytoclair), has been used to obtain a centrifuged cell deposit for the detection of malignant cells in sputum. In 178 sputum samples from 52 patients with primary lung cancer there was a slightly higher rate of detection with the mucolytic method than with the routine method: 54 positives from 27 patients compared with 48 positives from 24 patients. Morphological studies using the mucolytic agent indicated that a millipore filter with fixation of cells in suspension produces a satisfactory cell preparation and is valuable in certain problem cases.


A histological comparison of adenomatous and hyperplastic parathyroid glands. The histological findings in 18 cases of parathyroid hyperplasia associated with chronic renal failure and haemodialysis have been compared with a series of 35 cases of primary adenomatous hyperparathyroidism. Analysis of several features suggests that there are no definite criteria for distinguishing microscopically between individual enlarged glands in primary and secondary hyperparathyroidism, although nuclear pleomorphism is more common in primary adenoma and nodules are more common in secondary hyperplasia. These findings are discussed.


Myometrial vascular damage after surgical sterilisation by tubal diathermy. Vascular damage is described in the myometrium adjacent to areas of previous tubal diathermy for sterilisation. Elastosis, different in degree from that seen in the multiparous uterus, is most common, but more striking is medial arteriolar muscle loss with microaneurysm formation and occasional rupture.


Further characterisation of a spontaneously occurring antibasement membrane antibody. A spontaneously occurring antibasement membrane antibody has been further characterised according to antigenic specificity, immunoglobulin class, and tissue localisation using immunofluorescent and immunoelectron microscopic techniques. The autoantibody reacted with the basement membrane of kidney tubules, Bowman's capsule, and the epithelial portion of ileum but not with the basement membrane of skin, cornea, glomerulus, or oesophagus. It also reacted with bile canaliculi, sarcolemmal sheath, and salivary duct. On electron microscopy the antibody was distributed along the basement membrane of Bowman's capsule and renal tubules. Some reactivity against collagen was observed. Antibody activity was found in both IgG and IgM fractions. In immunodeficiency disorders, the autoantibody was found only in patients with selective IgA deficiency.


Output of peritoneal cells during peritoneal dialysis. Peritoneal dialysis provides a good source for the collection of macrophages. Six patients with chronic renal failure undergoing peritoneal dialysis for the first time were studied, and maximum cell egress, mostly macrophages, occurred at 24-48 hours and diminished after 48 hours.


Blood cells and alcohol consumption with special reference to smoking habits. In the course of the Paris study on risk factors of cardiovascular disease in a large professional group, 7710 active and apparently healthy men aged between 48 and 54 were examined. This study measured the relationship between clinical abnormalities suggesting alcoholic liver disease (ALD) and the following blood parameters: white (WBC) and red (RBC) blood cell counts, haematocrit (H), and mean corpuscular volume (MCV), the former computed as H/RBC ratio. A subsequent analysis was performed on a random sample of 485 subjects without ALD who were questioned on their daily average alcohol consumption. Each subject was classified as ;smoker' or ;non-smoker' according to his daily tobacco consumption for the last five years. Analysis of the data confirmed that smoking and alcohol were related to the blood parameters; but, according to smoking habit, different relationships between alcohol consumption (or ALD) and MCV, RBC, or WBC counts were found: for smokers, RBC count significantly decreased and MCV increased with alcohol consumption (or ALD); for non-smokers, WBC count significantly increased with alcohol consumption (or ALD). So, it would be of interest to consider the relation between alcohol and tobacco in interpreting possible changes in blood parameters and in formulating hypotheses on the mechanisms of their specific action.


Specific and non-specific folate binding protein in normal and malignant human tissues. Binding of tritiated folic acid by supernatants prepared from extracts of normal and leukaemic leucocytes, normal mucosa, and malignant tumours from different parts of the gastrointestinal tract has been measured using Sephadex-gel filtration and albumin-coated charcoal techniques. Non-specific binding (measured by Sephadex G-75 gel filtration) was almost invariably greater than specific binding measured by albumin-coated charcoal separation of bound and unbound folate. In nine normal leucocyte extracts, binding measured by Sephadex G-75 filtration ranged from 1.3 to 18.2 (mean 8.2) pg/mg protein and by albumin-coated charcoal from 1.0 to 14.8 (mean 6.7) pg/mg protein. Raised specific binding was found in the extracts from leucocytes of eight of 14 patients with chronic granulocytic leukaemia, in four substantially so (389, 121, 108, 59.7 pg/mg protein), but was only marginally increased in one of eight cases of acute myeloid leukaemia and in two of five cases of chronic lymphocytic leukaemia. Binding was normal in the extracts of all three cases of acute lymphoblastic leukaemia tested. Among the tissues of the gastrointestinal tract binding was greatest by the duodenal mucosa and liver. Extracts of carcinoma of the stomach and colon bound greater amounts of (3)H-folic acid than the corresponding normal mucosal extracts but the differences were not large. Sephadex G-200 gel chromatography showed more than one binding peak in the extracts of liver and duodenum but only one peak in the other tissues of the gastrointestinal tract, and only one peak, of molecular weight either about 50 000 or over 200 000, in the leucocyte extracts.


Urinary free corticosteroid excretion and renal function. Theoretically urinary free corticosteroid excretion should be affected by renal function and this would make it a less sensitive index of hypercortisolaemia. In 28 consecutive urine samples there was a clear relationship (r = 0.83; P less than 0.001) over a range of creatinine clearances 0.3-200 ml/min. Although an allowance could be made for renal function this would not necessarily improve the discrimination of normal from abnormal. Until data comparing corrected to uncorrected urinary free corticosteroid excretion become available, we recommend a short dexamethasone test as the initial investigation in patients with suspected hypercortisolaemia and abnormal plasma creatinine concentrations.


A computer-assisted bacteriology reporting and information system. A computer system for reporting and recording all specimens processed by the routine bacteriology laboratory at the Royal Postgraduate Medical School is described. Features of interest are the method of input using a mixture of 3-character alphanumeric codes and numbers, cumulative reporting to the wards, and selective listing of relevant previous results for the patient, which is available to technical and supervisory staff during processing of the specimen. The relative value to the wards and the laboratory of each type of information transfer has been assessed. Overall the use of a computer has resulted in higher quality bench work and more accurate reporting. It seems little more expensive than a previous manual system, although it has transferred work from the technical to the clerical staff.


Viral antigens and antibodies in hepatitis B infection. Hepatitis B virus antigens and antibodies were detected in the sera of acute and persistently infected patients. Evidence of active virus replication was confined to immediately before or during the initial detection of hepatitis B surface antigen during acute hepatitis B. Hepatitis B core antibody appeared during the period of antigenaemia and preceded recovery. Hepatitis B e antigen was dound in a proportion of sera which contained significant levels of virus particles. In contrast, all sera containing hepatitis B virus particles from persistently infected patients treated by maintenance haemodialysis also contained the e antigen. Among a group of 50 persistent carriers of hepatitis B virus, significant levels of virus production occurred in the presence of antibody to e antigen. In addition, evidence of exposure to hepatitis B virus was found among 3% of blood donors in whose sera the surface antigen was not detected by radioimmunoassay. The significance of these findings is discussed in relation to the aetiology of hepatitis type B.


A cerebrospinal fluid leucocidin in pyogenic meningitis. Cerebrospinal fluid (CSF) samples were tested for their cytotoxicity to polymorphonuclear neutrophil leucocytes (PMN) using a 51Cr release assay. Most samples from patients with pyogenic meningitis damaged PMN while normal CSF samples did not. No difference was found between the cytotoxic activity of CSF from patients with pneumococcal meningitis and from patients with meningococcal meningitis. It is, therefore, unlikely that a CSF leucocidin plays an important part in producing the high mortality of pneumococcal meningitis.


Chlorthalidone-induced syndrome of inappropriate secretion of antidiuretic hormone. A 60-year-old woman who had been instructed to increase her water intake because of nephrolithiasis developed the syndrome of inappropriate secretion of antidiuretic hormone when treated with chlorthalidone for mild hypertension. Serum osmolality was 235 mOsm/kg with concomitant urine osmolality of 490 mOsm/kg. When serum sodium decreased to 110 mEq/liter, plasma antidiuretic hormone (ADH) was elevated at 30 pg/ml. The syndrome resolved when chlorthalidone was discontinued together with fluid intake restriction. Plasma ADH returned to normal (less than 0.5 pg/ml) after three days of treatment. The favorable outcome in this patient is attributed to early recognition of the syndrome, which might occur even with nonthiazide diuretics such as chlorthalidone.


Syntax influences the accuracy of consonant production in language-disordered children. Nine children with restricted syntactic and phonological development repeated 20 target consonants embbedded in noun phrase, simple declarative, and passive structures to determine the influence of syntactic complexity on accuracy of consonant production. The children made significantly more errors in the sentence contexts than in the noun phrase context. This was found for both early-developing and late-developing consonants. However, the ratio of the number of distinctive feature modifications to the number of consonant errors showed that segmental errors were not produced as less exact approximations of the target consonants. Instead, the children simply made more errors of a predictable type, suggesting that the effect of syntax on accuracy of consonant production is quantitative rather than qualitative.


Testing for a specific syntactic structure. Nine language delayed children who had failed the "is interrogative" test item on the Programmed Conditioning for Language Test served as subjects. A comparison was made of subjects' syntactic performance on six language tasks designed by the investigators to elicit the "is interrogative" structure. These tasks ranged along an imitative-to-spontaneous continuum demonstrating varying degrees of communicative intention, presence or absence of contextual referents, and varying levels of task structure. Analyses indicated all nine subjects produced complete grammatical "is interrogative" utterances on one or more of the language tasks. The most effective tasks for eliciting the specific structure were those that required communicative intention, included specific contextual referents, and provided a relatively structured situation. The direct imitative task was less effective, although it elicited more correct productions than the spontaneous language sampling task. However, the relative eliciting potential of each task was not the same for each child. Theoretical and clinical implications of these findings are discussed.


The effects of pause time on auditory comprehension of language-disordered children. Language-disordered children were presented with two types of commands in a sentence comprehension task. One 3-sec pause was placed at the same syntactic boundary in one-half of the sentences in Set I and Set II. Results indicate that the subjects' performance improved as a function of the pause in Sentence Set II but not in Sentence Set I.


Discrimination of speech intensity differences via vibrotactile stimulation. This study investigated vibrotactile difference limen judgments for intensity using certain speech signals as stimuli. The variables under investigation were those of stimulus presentation level, direction of intensity increment or decrement within each paired-comparison stimulus complex, and the spectral composition of the speech signals. The results indicate that the level of stimulus presentation did not affect difference limen decisions among any of the sensation levels utilized. The largest most consistent effect upon difference limen judgments was in the direction of intensity change. The difference limen judgments were smaller when the second stimulus of the pair was of less intensity than the first. There was no effect upon difference limen decision as the result of stimuli having different spectral compositions.


On the comprehension of questions by nonspeaking children. The question form presents both a semantic and linguistic problem to deaf and other language impaired children. The wh question requires a pronominal reference system to be well-established before the wh question can be comprehended and produced. The present study describes the language competence of a deaf, partially sighted girl who acquired language by reading it. This child neither hears nor speaks. Communication with her family was accomplished by writing in her hand, gestures, and pictures. The present study explores pronominal reference in relation to wh questions. A method of teaching was devised to establish a system of pronominal reference, which enabled this adolescent girl to comprehend and produce wh questions.


Intensity coding in the transient portion of the rat chorda tympani response. For assessment of the relative importance of the transient and steady state components of the chorda tympani response to intensity coding of gustatory stimuli, the rat's ability to discriminate aversive solutions following adaptation was compared with changes in the neural response also resulting from adaptation. A taste aversion was conditioned to .9% NaCl, and subsequent acceptance of several NaCl concentrations (range: .1%-1.5%) was tested following infusion of 1 ml of adapting solution (H2O or .1%, .9%, or 1.5% NaCl) through an intra-oral cannula. Adaptation significantly decreased gustatory intensity discrimination and decreased the magnitude of the transient portion of the chorda tympani response. The degree of reduction in transient magnitude correlated significantly with reduced discrimination in all adapting conditions. However, the magnitude of the steady state response negatively correlated with discrimination. These data suggest that gustatory intensity discrimination depends upon information contained in the transient portion of the neural response and that the transient is necessary as well as sufficient for taste intensity discrimination.


Abdominal vagotomy disrupts food-related drinking in the rat. Rats with complete subdiaphragmatic bilateral transection of the abdominal vagus (Vgx-C) showed disordered food-related drinking when drinking water in temporal association with a meal of dry food after 5-hr food deprivation and when drinking water in association with a liquid meal after 24-hr food deprivation. The Vgx-C rats drank after significantly longer latencies and drank significantly less water in 1 hr than did sham-vagotomized (Sham) rats after eating the same size meal (solid or liquid) as Shams. Rats with incomplete vagal transection (Vgx-I) ate and drank like Shams. Water intake of Sham and Vgx-I rats correlated positively with the meal size of solid food, but the water intake of Vgx-C rats did not. The failure of Vgx-C rats to drink water normally when food was ingested was not due to failure of a food stimulus to reach the intestine, because Vgx-C and Sham rats emptied equivalent volumes of liquid food from the stomach into the intestine within 10 min of food entering the stomach. These results indicate that the abdominal vagus is an important neurological substrate for food-related drinking in the rat.


Antidipsogenic role of the E-prostaglandins. Prostaglandin E1 (PGE1) is antidipsogenic when administered into the lateral cerebral ventricle of the rat. In these experiments PGE1, at a dose of 1 microgram, suppressed water intake induced by centrally administered angiotensin II (AII) or carbachol, subcutaneously administered polyethylene glycol, and water deprivation. Even at this high dose, PGE1 did not reduce water intake due to cellular dehydration. As the dose of PGE1 was reduced, its suppressant effects became more specific to AII-induced drinking, with a dose of 10 ng yielding a significant suppression only in the case of AII-induced drinking. Even at a high dose (1 microgram), the PGE1 suppression of ingestion was specific to water intake. Although PGE1-treated rats reduced food intake if they were not hydrated prior to access to food, no reduction in food intake was seen when they received water by gavage before food presentation. Since PGE-like substances are synthesized in the brain, these data suggest the possibility that the prostaglandin may be involved in the regulation of water balance as a component of a reciprocal system in which water intake initiated by the activation of the renin-angiotensin system is halted by the synthesis and release of PGE.


Huddling by rat pups: multisensory control of contact behavior. Huddling, or clumping behavior, begins in the litter situation and persists throughout the adult life of the rat (Rattus norvegicus). In pups, as in adults, the behavior provides a metabolically important means of reducing heat loss. The sensory stimuli that elicit, direct, and maintain huddling in developing rat pups (5-20 days of age) were studied in a standardized testing situation by using long-term time-lapse video observations. All pups huddled with a single, immobile sibling. A loop of warm tubing also elicited huddline. However, it appears that other, nonthermal cues can elicit huddling, since a dead, ambient-temperature pup was a sufficient stimulus for the behavior. Disruption of normal olfactory input by intranasal infusion of zinc sulfate interfered with the maintenance of contact. In addition, two kinds of tactile stimulus, vertical contours and furry "comfort" cues, as well as photic responses appeared to participate in the mediation of huddling. Huddling in the altricial rat is under multisensory control, although the number of cues used by pups for huddling appears to increase with age.


Huddling by rat pups: group behavioral mechanisms of temperature regulation and energy conservation. Body heat loss was attenuated and oxygen consumption was reduced by huddling in litters of developing rats. Rat pups derive physiological benefits from huddling similar to those enjoyed by adult mammals; these findings contrast with previous characterization of the altricial rat as poikilothermic. Huddling insulates by lessening the explosed body surface area of the participants, thus retarding heat loss and enhancing the efficiency of thermogenesis. These physical mechanisms of the clump are actively regulated by the pups. A novel quantitative measure of huddle size revealed a form of group regulatory behavior in rat pups whereby the surface expanded and contracted with increases and decreases in ambient temperature. The individual basis of this group regulatory activity was investigated by marking individual pups and observing them in huddles by means of time-lapse videography. It was found that individual animals circulate throught the huddle, frequently exchanging locations in the group. By studying the huddle positions of an anesthetized pup and a marked control sibling, dynamics of the pup flow were clarified. Ordinarily, the direction of movement was actively downward, into the pile; immobile pups "floated" on the surface. When the nest temperature was raised to thermoneutral, the direction of pup flow reversed and an immobile animal sank to the depths of the huddle. Through individual competitive adjustments the huddle behaves as a self-regulating unit which provides warmth and insulation to all its active members.


Extent of recovery from neonatal damage to the cortical visual system in cats. Cats that received either marginal or marginal plus extramarginal lesions as 3-day-old kittens were assessed on a series of tests of visually guided behavior. These cats were not conspicuously different from normal controls in avoiding obstacles or in activity level. Yet these same operated cats were severely impaired in performance on the visual cliff and in visual discrimination learning, even when the lesions were limited to the geniculocortical portion of the visual system. However, maximum losses in pattern and form discrimination learning were observed only in cats with severe retrograde degeneration in both the lateral geniculate nucleus and the complex of the pulvinar and nucleus lateralis posterior. Photically evoked potentials were recorded in the lateral regions of the neocortex more reliably from operated cats that had made fewer errors in discrimination learning than from more severely debilitated cases; this relation was present even among cases with nearly equivalent amounts of retrograde degeneration in the visual thalamus. These findings suggest that in the cat (a) recovery of vision is incomplete after neonatal lesions of the visual cortex and (b) a cortical system lateral to the geniculocortical projections may be involved in pattern vision.


Relation of albinism and drugs to the visual evoked potential of the mouse. The individual and combined influences of pentobarbital and chlorprothixene on the early components of the cortical visual evoked potential (VEP) were examined in the C57BL/6 mouse. Pentobarbital produced a large increase in latency, and chlorprothixene resulted in a smaller latency increase. When these agents were combined, their effects on the VEP latency were antagonistic. The effects of pentobarbital on VEP amplitude varied as a function of flash intensity, and this barbiturate anesthetic also produced qualitative VEP waveform changes. When the congenic albino (c/c) C57BL/6 was compared with the black (+/+) C57BL/6 mouse; pentobarbital anesthesia produced a decrease of latency with increasing stimulus intensity only in the c/c genotype, whereas VEP amplitudes were similarly affected by pentobarbital in both genotypes.


Disappearance of hoarding and disorganization of eating behavior after ventral mesencephalic tegmentum lesions in rats. The effects of ventral mesencephalic tegmentum (VMT) radio-frequency lessions on spontaneous and complex behaviors, such as food hoarding and alimentary patterns, were investigated. Activity measures indicated that VMT lesions increased horizontal activity in a circular corridor as well as in an open field whereas vertical activity (rearing) was decreased. After 12 hr of food deprivation, food hoarding and alimentary patterns were evaluated in a square open field to which rats had free access from their home cage. Control rats hoarded food and had organized feeding behavior, but experimental rats did not hoard. Moreover, increasing the open-field illumination did not reverse this deficit. Similarly, the VMT lesion induced disruption of the adaptative alimentary behavior in response to food deprivation. A strong correlation was observed between hoarding scores and the extent of locomotor hyperactivity. These results are discussed in terms of a possible implication of limbic-midbrain mechanisms in which the meso-cortico-limbic dopaminergic A10 neurons originating in the VMT might have a primary role.


Effects of context manipulation on memory and reversal learning in rats with hippocampal lesions. In Experiment 1, groups of hippocampal and control rats learned a simultaneous discrimination habit and were subsequently tested for its retention under variable contextual conditions. All groups recalled the discrimination response to an equally high level when testing conditions were constant throughout, but the hippocampal group showed impaired memory when contextual stimuli at recall testing did not conform to those of original learning. Experiment 2 indicated that the hippocampal impairment was not simply the result of introducing novel stimuli. In Experimental 3, animals were administered a reversal learning task with contextual stimuli varied between the two tests. The typically observed impairment of hippocampal animals on this task was reduced by contrasting contextual conditions. The data are seen to support a contex-tetrieval interpretation of hippocampal function.


Emerging cholinergic mechanisms and ontogeny of response inhibition in the mouse. Mice, 7, 11, 15, 19, and 85-115 (adult) days of age, served as subjects in experiments assessing effects of anticholinergics on the development of behavioral inhibition. The centrally active anticholinergic scopolamine produced a dose-dependent elevation in locomotor activity in 19-day-old and adult mice. Acquisition and retention of a step-off passive avoidance response (PAR) was initially studied in nondrugged subjects. Mice as young as 7 days of age learned and retained the PAR for 1 hr. Twenty-four-hour savings, however, were not observed until 19 days of age. Simple PAR performance deficits following scopolamine injection were first seen at 15 days of age. Mice in those age groups exhibiting 24-hr retention (19-day-olds and adults) were used to assess carry-over effects of scopolamine on retest. Only in the case of juveniles did scopolamine, injected prior to training, disrupt 24-hr retest performance. Since methylscopolamine, a peripherally active anticholinergic, had no effect on activity and PAR performance, it is assumed that scopolamine's effects were of central origin. The results suggest that behavioral suppression comes under cholinergic control during the second and third postnatal weeks but that cholinergic mechanisms may not mediate response inhibition uniformly throughout development.


Osmoregulatory thirst in rats after lateral preoptic lesions. In confirmation of previous observations, lesions of the lateral preoptic area in rats abolished water ingestion during a 4-hr test following an intraperitoneal injection of hypertonic NaCl solution. However, it was observed that these animals increased their water intakes when tests were prolonged to 24 hr and drank almost exactly what they needed for osmoregulation. Furthermore, they increased their water intakes normally when NaCl was given in their diet, when NaCl was administered intravenously, or when they were water deprived and given preloads of isotonic saline to remove hypovolemia. These findings indicate that rats with lateral preoptic lesions do experience osmoregulatory thirst, and consequently, they raise new doubts about whether osmoreceptors located in the lateral preoptic area mediate thirst following the administration of osmotic loads.


Garter snake trailing behavior: effects of varying prey-extract concentration and mode of prey-extract presentation. In a multiple-choice maze, garter snakes were trained to follow earthworm-extract trails for worm bit rewards. In Experiment 1, they were tested for their abilities fo follow extract trails that had been dried or extract trails that were removed from direct lingual access by a perforated floor. Snakes were able to follow the dry trails and unable to follow removed trails. In Experiment 2, snakes were tested for their behavioral responses to different concentrations of extract trails. Snakes trailed more accurately, moved more slowly, and exhibited much higher tongue flick rates on the intense concentration trails. The results are interpreted in terms of the assumption that effective trails are perceived by the tongue flick delivery of odorants to the vomeronasal organs.


Computer assisted tomography of normal suprarenal glands. The anatomy of 60 right and 80 left normal suprarenal (adrenal) glands in 90 patients was studied. In the transaxial view, the glands have a triradiate shape, which varies on serial sections. The medial posterior limbs of each gland were visualized on the more cranial sections. The transaxial positions of the glands are related to the vertebrae and, on the left side, to the aorta and kidney, in accordance with known anatomical associations. The cranio-caudal positions of the glands have been defined by using the kidney as a "renal ruler". The right gland lies cranial to the anterior and medial renal margin, and it extends dorsally beyond the vertebral body in 18% of the right glands studied. The left gland lies anterior to the upper renal pole. The ease with which the glands were shown in detail by computer assisted tomography (CAT) contrasts with other methods of demonstration, namely, retroperitoneal pneumography, angiography, and venography. The difficulties of visualizing the glands with CAT are discussed and illustrated. The findings are used to suggest guidelines for conventional tomography.


A method for correcting bone induced artifacts in computed tomography scanners. A method that corrects for artifacts in X-ray transmission scanners created by the alteration of the energy spectrum by bone is described. The method involves two reconstructions of the pictures: the first establishes the approximate distribution of bone and the second enables the artifacts to be eliminated. The first reconstruction is needed to establish the total amount of bone along each ray. Assuming that the X-ray energy spectrum is known, it is shown how the line integrals can be corrected. The method was tested on several mathematical phantoms as well as on human head data. The nonlinear nature of the spectral artifacts is shown to produce streaks in certain phantoms.


Anthropomorphic phantoms for computed tomography scanner performance evaluation. Phantom simulating materials and geometries encountered in computed tomography of the head and body have been constructed. They permit the quantitative study of artifactual behavior as well as low contrast detectability and spatial resolution of a computed tomography (CT) scanner. These phantoms may be useful in comparative evaluation of CT performance and in quality control programs.


Adrenal cortical tumors with low attenuation coefficients: a pitfall in computed tomography diagnosis. Four cases of adrenal tumors with unusually low attenuation coefficients are reported. The tumors presented with values ranging from 8 to 17 EMI units (EU: 500 scale) instead of the expected 30 to 40 EU range of the remainder of our adrenal tumors. Two of these patients had primary aldosteronism and two had Cushing's syndrome. An unusually high lipid content may be responsible for the lucent appearance.


The base of the skull: a comparison of computed and conventional tomography. The diagnostic information obtained from 35 examinations of the skull base by computed tomography (CT) was compared with the findings furnished by conventional or blurring tomography (BT). Computed tomography yielded diagnostically valuable information not only on the soft tissues but also on the bony structures. Compared with BT, CT was superior in 40%, equivalent in 40%, and inferior in 20% of the cases in assessing pathological findings involving the bony structures of the skull base. Obviously, CT's additional capability of accurate display of soft tissue changes represents a fundamental diagnostic advantage.