Influence of anesthetics, ethyl alcohol, and Freon on dark adaptation of monkey cone ERG.

Cone dark adaptation curves were measured in a rhesus monkey using the electroretinogram (ERG) response to a 40 Hz flickering stimulus. The influence of anesthetics on the time course of dark adaptation was studied. All volatile anesthetics tested (methoxyflurane, halothane, enflurane, ether, chloroform) retarded dark adaptation but to different degrees; urethane, ethyl alcohol, and Freon 11 also retarded dark adaptation. No effect was found for barbiturates and ketamine. It seems unlikely that metabolites play a role in the observed phenomena. A literature survey reveals that several studies on dark adaptation or visual pigment regeneration might have suffered from influences of the anesthetic used. The cause of the phenomenon might lie either in anesthetics-induced membrane changes or in hindrance of the isomeration of 11-trans retinal to 11-cis retinal.


Displaced ganglion cells in the retina of the monkey.

Horseradish peroxidase has been injected into the dorsal lateral geniculate nucleus of macaque monkeys to label ganglion cells of the retina by retrograde axoplasmic transport. Displaced ganglion cells, with somata in the inner nuclear layer and dendrites in the inner plexiform layer, were detected by virtue of their filling with peroxidase-positive granules. These cells were numerous in the peripapillary region, but relatively uncommon elsewhere in the retina.


Evaluation of a new gonorrhea culture detection system--Microcult G.C.

A new dehydrated test system (Microcult G.C.), developed by the Ames Company for the detection of Neisseria gonorrhoeae, was evaluated with stock cultures and fresh isolates. When evaluated against fresh, modified Thayer-Martin (MTM) medium, the new test was slightly less sensitive (81.6% vs 84.2% detection); however, Microcult G.C. has the unique advantage of long room-temperature storage life without deterioration. Thus, it would probably not be used where fresh MTM is readily available, proving most useful in areas where laboratory services are less accessible.


Neisseria gonorrhoeae identification in carbohydrate medium containing guinea pig serum.

Clinical isolates of Neisseria gonorrhoeae from Manitoba were tested for their carbohydrate degradation activity in Cystine Trypticase Agar (C.T.A) medium, Mueller-Hinton Agar and guinea pig serum agar. Each isolate was tested using 2 carbohydrates (e.g. glucose and maltose) in the above three media. Out of 661 isolates tested, only 80% were positively identified in C.T.A. medium. Mueller-Hinton agar allowed 88% identification while guinea pig serum agar yielded 100% identification. In a second series of experiments, 102 cultures of N. gonorrhoeae were used to compare Flynn & Waitkins medium with guinea pig serum agar. Only 91 of these were identified with Flynn and Waitkins medium while guinea pig serum agar identified all the 102 isolates. Guinea pig serum provides adequate growth of fastidious N. gonorrhoeae essential for detecting specific enzymes. Since guinea pig serum does not contain maltase activity, it does not interfere with the biochemical activities tested. Guinea pig serum agar is easy to prepare, does not require a heavy inoculum and gives definite color change in the medium.


An epizootiologic study of anthrax in Falls County, Texas.

In June and July, 1974, an anthrax epizootic in Falls County, Texas, resulted in the death of 236 animals (228 cattle, 5 horses, 2 mules, and 1 pig) on 48 premises. Death rates were highest for horses (18.2%) and bulls (16.8%). The epizootic was apparently precipitated by drought, and infection appeared to be the result of ingesting intrinsically contaminated soil and grass. Human illness was not associated with the epizootic.


High pressure liquid chromatographic determination of 4,4'-(diazoamino)-dibenzenesulfonic acid in FD&C yellow no. 6.

Fourteen analysts from 9 laboratories evaluated a high pressure liquid chromatographic procedure for the determination of 4,4'-(diazoamino)-dibenzenesulfonic acid (DAADBSA) in FD&C Yellow No. 6. Each collaborator analyzed 5 samples nominally containing 0-0.050% DAADBSA. The repeatability and reproducibility of the method are estimated to be 0.003% and 0.020%, respectively. The method has been adopted as offical fist action.


Analysis of polychlorinated biphenyls by carbon-13 nuclear magnetic resonance spectroscopy.

A technique is presented for the analysis of polychlorinated biphenyl mixtures by assigning 13C resonances to the observed peaks and matching these shifts to the shifts of known individual polychlorinated biphenyls. The technique has not proved to be as useful for heavily chlorinated mixtures.


Structure of tetanus toxin. I. Breakdown of the toxin molecule and discrimination between polypeptide fragments.

Tetanus toxin was digested with papain, yielding one major polypeptide (Fragment C) with a molecular weight corresponding to 47,000 +/- 5%, thus comprising about one-third of the toxin molecule. Fragment C was antigenically active, atoxic, and stimulated the formation of antibodies neutralizing the lethal action of tetanus toxin in vivo. Furthermore, a second split product (Fragment B) was isolated from the papain digest, containing two polypeptide chains linked together via a  disulfide bond. Fragment B (Mr = 95,000 +/- 5%) was atoxic and showed a reaction of nonidentity with Fragment C on immunodiffusion analysis against tetanus antitoxin. The basic two-chain structure (heavy and light chain polypeptide, cf. Matsuda, M., and Yoneda, M. (1975) Infect. Immun. 12, 1147-1153) of tetanus toxin has been confirmed and the relationship between Fragments B and C within this framework has been established. Fragment C was distinguished from the light chain by electrophoresis in sodium dodecyl sulfate and by immunodiffusion analysis, indicating that this fragment constitutes a portion of the heavy chain polypeptide. Fragment B showed a reaction of partial identity with the light as well as the heavy chain from tetanus toxin. Reduction of Fragment B with dithiothreitol followed by gel chromatography yielded a fraction which was indistinguishable from the light chain portion of the toxin molecule. It is concluded that Fragment B comprises the complementary portion of the heavy chain (remaining after scission of the polypeptide bond(s) releasing Fragment C) linked to the light chain by a disulfide bond.


Nitrogen 15 tracer studies on the pathway of denitrification in Pseudomonas aeruginosa.

The pathway of anaerobic reduction of nitrite to nitrogen gas (N2) by cell suspensions of the denitrifier, Pseudomonas aeruginosa, was studied using the techniques of gas chromatography and mass spectrometry. While release of nitrous oxide (N2O) is not normally detected during the reduction of nitrite to N2 by this organism, 15N from [15N]nitrite nevertheless can be trapped quantitatively as 15N2O in a pool of added N2O. In such experiments the abundance of 15N in N2O always exceeds that in product N2, consistent with the absence of a major reductive route from nitrite to N2 which by-passes N2O. During the reduction of a mixture of [15N]nitrite and nitric oxide (NO), 15NO produced at most only in trace amounts. The final products are chiefly 15N2 and 14N2 with only a small fraction of the scrambled product, 14N15N. Much of the 14N15N can be accounted for as an artifact caused by traces of molecular oxygen, which promote the conversion of NO to nitrite by autooxidation and thereby degrade slightly the isotopic purity of [15N]nitrite. Nitrous oxide shows all the properties of a free obligatory intermediate during the denitrification of nitrite to N2 by P. aeruginosa, whereas NO does not. The inability to trap 15NO in a pool of NO indicates that NO is not a free obligatory intermediate in the reduction of nitrite. The small mole fractions of 14N15N produced from a mixture of [15N]nitrite and NO require that the main reductive pathways for these nitrogen oxides cannot share any freely diffusible mono-nitrogen intermediate in common. The simplest interpretation is that nitrite and NO are denitrified by separate pathways, at least prior to the formation of the first bi-nitrogen compound.


A dimer of a single polypeptide chain catalyzes the terminal four reactions of the L-tryptophan pathway in Euglena gracilis.

In Euglena gracilis the terminal four enzyme activities of the tryptophan biosynthetic pathway were found to be associated with a protein with an estimated molecular weight of 325,000 +/- 20,000. The protein was purified approximately 2,000-fold with relatively proportional recoveries of all four enzyme activities. The purified material was homogeneous by the criteria of analytical disc gel electrophoresis and gel isoelectric focusing. Disc gel electrophoresis after denaturation with sodium dodecyl sulfate gave a single protein band with a molecular weight of 155,000 +/- 5,000. Disc gel electrophoresis in 8 M urea also gave rise to a single protein band. We interpret these results as evidence for a single species of subunit. The pathway in Euglena is the only one known to the present in which the terminal enzyme, tryptophan synthase, is not a separate molecular species.


New types of RNA polymerase mutations causing temperature-sensitive sporulation in bacillus subtilis.

A single site mutant of Bacillus subtilis with a streptovaricin-resistant RNA polymerase has been isolated; this mutation caused temperature-sensitive sporulation, but had no effect on vegetative growth. The mutant (ts710) temperature-sensitive period irreversibly affected the middle and late stages of sporulation. Mutant cells grown at the nonpermissive temperature exhibited abnormal serine protease accumulation, serine esterase accumulation, alkaline phosphatase accumulation, RNA polymerase template specificity changes, and pulse-labeled RNA synthesis profiles. The accumulation of metal protease was not affected at the nonpermissive temperature. Attempts to isolate single site mutants which were streptolydigin-resistant, and temperature-sensitive for sporulation, were unsuccessful.


Development of sarcoplasmic reticulum in cultured chicken muscle.

The development of sarcoplasmic reticulum membranes was studied in vivo and in tissue culture in chicken pectoralis muscle cells. The concentration of the calcium- and magnesium-activated ATPase measured by selective labeling of the enzyme with [32P]ATP in whole muscle homogenates was found to increase in developing chicken pectoralis muscle in vivo from 0.01 nmol/mg of protein in 12-day embryos to 0.3 to 0.4 nmol/mg of protein in 1-month-old chicks, where it constitutes about 3% of the total protein content of muscle. In cultured muscle cells the concentration of calcium-sensitive phosphoprotein increased from 0.015 nmol/mg of protein at 2 days to 0.04 to 0.05 nmol/mg of protein after 5 days of culture. This amount represents about 0.5% of the protein content of the muscle cells. The accumulation of Ca2+ transport ATPase began during fusion and continued with a linear rate during 8 days of culture. The density of 75 A intramembranous particles seen by freeze-etch electron microscopy on fracture faces of sarcoplasmic reticulum membranes is about 4,000/mum2 in adult chick pectoralis muscle but only 400/mum2 in cultured muscle cells in rough proportion to the concentration of Ca2+-sensitive phosphoprotein. The Ca2+, Na+, and K+ concentration of the medium and addition of ouabain, caffeine, or the calcium ionophores A23187 and X537A sharply influence the concentration of calcium transport ATPase in cultured muscle cells, parallel with their effect upon cell fusion and growth. These observations are consistent with the proposition that the gene expression leading to the accumulation of Ca2+ transport ATPase during development in culture may be regulated by intracellular ion concentrations.


Microtubular proteins of Chlamydomonas reinhardtii. An immunochemical study based on the use of an antibody specific for the beta-tubulin subunit.

An immunochemical assay for tubulin subunits is described. The method is applied directly to homogenates of Chlamydomonas reinhardtii solubilized in sodium dodecyl sulfate (Na dodecyl-SO4), and it makes use of a two-dimensional electrophoresis system; the first separation is carried out by Na dodecyl-SO4-polyacrylamide gel electrophoresis and the second by electrophoresis into an agarose gel containing antibodies. Tubulin is precipitated in the form of a "rocket" and the method is made quantitative through the use of cells labeled with [35S]sulfate. The antiserum used in this assay was prepared in rabbits using beta subunit of tubulin purified from Chlamydomonas flagella by two preparative Na dodecyl-SO4-polyacrylamide gel electrophoreses. This antiserum and an antiserum to alpha subunit of tubulin from porcine brain, prepared for comparative study, were extensively characterized. Both antisera show specificity for the polypeptide used as antigen and react with the native dimeric tubulin. The antiserum to beta subunit from Chlamydomonas flagella also forms immunoprecipitates with native brain tubulin and its beta subunit when used at high titer. In contrast, the antiserum to alpha subunit from porcine brain does not cross-react with Chlamydomonas tubulin. The immunochemical assay was applied to Chlamydomonas cells synchronized by a 12-h light/dark cycle. In cells collected during the light period (late G1), after removal of flagella, the content of tubulin is estimated to be 0.3% of total protein. As cells enter the dark period there is a striking increase in tubulin content which reaches a maximum just before cell division.


Activation of rat adipocyte glycogen synthase by insulins.

Incubation of fat cells with insulin increased glycogen synthase I activity without changing total synthase activity. This effect of insulin was dependent upon the particular lot of albumin present in the medium and was abolished by incubating cells with trypsin. Half-maximal activation of glycogen synthase was obtained with 8 microunits/ml of insulin, a concentration very similar to that which half-maximally stimulated 3-O-methylglucose uptake. The basal percentage of phosphorylase a activity was not detectably altered by insulin, although it was decreased by incubating cells with 5 mM glucose. Insulin (50 microunits/ml) markedly opposed actions of epinephrine (0.05 to 10 muM) to increase phosphorylase a activity and decrease glycogen synthase I activity, effects which were observed without glucose. Partial activation of glycogen synthase by insulin was seen after 1 min and complete activation after 4 min. Glucose alone produced a transient increase in synthase I activity. When cells were incubated with insulin plus glucose for 4 min, the increase in the percent synthase I activity was much greater than the additive effects of insulin and glucose alone. This potentiation of the effect of insulin on glucogen synthase I activity depended on the time of incubation with glucose and on the concentration of the hexose. If  cells were incubated with cytochalasin B before insulin plus glucose, the effect of glucose was abolished. These results suggest that there are at least two mechanisms by which insulin can increase fat cell glycogen synthase I activity. One requires glucose and activation occurs secondary to an increase in glucose transport; where another mechanism(s) is operative even in the absence of glucose.


Purification and properties of a polymorphic high activity equine erythrocyte carbonic anhydrase.

A polymorphic form of the high activity or C-type of horse erythrocyte carbonic anhydrase has been isolated. It has been designated C2 and differs from the usual C1 form by having a cysteine replacement for arginine at residue 180. This second cysteine, unlike the other, is highly reactive. Isolation of the C2 isozyme by the usual methods results in most of it forming a mixed disulfide with glutathione and this product designated as C3 has an increased anodic mobility. The enzymatic activity and immunologic reactivity of both the C2 and C3 components are the same as for the usual C1 form of the enzyme. The C2 form can be stabilized by alkylation and the carboxamidomethyl derivative has been isolated in crystalline form.


Interspecies comparison of cytosolic and mitochondrial aspartate aminotransferases. Evidence for a more conservative evolution of the mitochondrial isoenzyme.

The degree of structural similarity between the mitochondrial isoenzymes of aspartate aminotransferase from pig heart and chicken heart was determined by means of their immunological cross-reactivity and compared with the degree of similarity between the cytosolic isoenzymes from the same two species. Quantitative microcomplement fixation revealed a remarkable similarity of the two mitochondrial isoenzymes corresponding to an immunological distance of 104. The structures of the two cytosolic isoenzymes, on the other hand, diverge with an immunological distance of 203. The apparent conservatism of mitochondrial aspartate aminotransferase indicates additional evolutionary constraints on the structure of this organelle-confined isoenzyme.


Purine nucleoside phosphorylase of rabbit liver. Mechanism of catalysis.

Initial velocity studies and product inhibition patterns for purine nucleoside phosphorylase from rabbit liver were examined in order to determine the predominant catalytic mechanism for the synthetic (forward) and phosphorolytic (reverse) reactions of the enzyme. Initial velocity studies in the absence of products gave intersecting or converging linear double reciprocal plots of the kinetic data for both the synthetic and phosphorolytic reactions of the enzyme. The observed kinetic pattern was consistent with a sequential mechanism, requiring that both substrates add to the enzyme before products may be released. The product inhibition patterns showed mutual competitive inhibition between guanine and guanosine as variable substrates and inhibitors. Ribose 1-phosphate and inorganic orthophosphate were also mutually competitive toward each other. Other combinations of substrates and products gave noncompetitive inhibition. Apparent inhibition constants calculated for guanine as competitive inhibitor and for ribose 1-phosphate as noncompetitive inhibitor of the enzyme, with guanosine as variable substrate, did not vary significantly with increasing concentrations of inorganic orthophosphate as fixed substrate. These results suggest that the mechanism was order and that substrates add to the enzyme in an obligatory order. Dead end inhibition studies carried out in the presence of the products guanine and ribose 1-phosphate, respectively, showed that the kinetically significant abortive ternary complexes of enzyme-guanine-inorganic orthophosphate (EQB) and enzyme-guanose-ribose 1-phosphate (EAP) are formed. The results of dead end inhibition studies are consistent with an obligatory order of substrate addition to the enzyme. The nucleoside or purine is probably the first substrate to form a binary complex with the enzyme, and with which inorganic orthophosphate or ribose 1-phosphate may interact as secondary substrates. The evidences presented in this investigation support an Ordered Theorell-Chance mechanism for the enzyme.


Carchesium stalk fibrillar matrix as a highly filled polymer network.

Glycerolated stalks of the sessile peritrich ciliate Carchesium sp. were treated with 10(-6) g ion/1 Ca2+ to disrupt the contractile spasmoneme. The resulting preparation consisted primarily of the fibrillar matrix, a dense extra-cellular meshwork of microfibrils. Some mechanical properties of this preparation have been investigated. The matrix tensile force-extension ratio relation for an initial stretch was characteristic of a soft, swollen polymer network, elastic modulus in young stalks 1.7 X 10(5) Nm-2, in mature stalks 4.0 X 10(5) Nm-2. The higher elastic modulus in mature stalks implies an increase in the interchain cross-link frequency. In young stalks, elastic modulus was found to be independent of the ambient Ca2+ concentration in the threshold range for spasmonemal contraction. Stalk relaxation was pronouncedly irreversible, showing stress softening and permanent hysteresis on repeated loading. Hysteresis was time independent and stiffness was not recovered after four hours at zero strain. Hysteresis was enhanced by repeated loading to the same tensile force. Stress-strain hysteresis at a low extension is characteristic of highly filled polymer networks in which polymer chains are interconnected via rigid filler particles as well as directly cross-linked.


Analysis of the proteins in sweat and urine by agarose-gel isotachophoresis.

A sensitive and practical method is described for the analysis of the proteins contained in human sweat and urine which does not require pre-concentration of the sample. Technical details are provided of the agarose-gel isotachophoresis and the proteinograms of normal and pathological urine samples, as well as proteinograms of human sweat. The method can also be applied as an electro-concentration system in a field-strength gradient. By means of this electro-concentration system, and in combination with immunodiffusion against monospecific antisera, a detection limit of albumin of 50 ng/ml has been obtained.


Low serum triiodothyronine (T3) and hypothyroidism in anorexia nervosa.

Measurements of serum thyroid hormones were compared in 22 patients with typical anorexia nervosa and 22 euthyroid control subjects. Serum total triiodothyronine (T3) was (mean +/- (SE) 62.1 +/- 7.1 ng/100 ml in anorexia patients and 115.2 +/- 8.4 ng/100 ml in control subjects (P less than 0.001). Serum adjusted thyroxine (T4Adj) was significantly different in the anorexia (7.1 +/- 0.4) and control (8.2 +/- 0.4) groups. Serum T3 was subnormal in 63% and T4Adj subnormal in 36% of the 22 anorexia patients. The mean serum T4/T3 in anorexia patients (158 +/- 19) was higher than that in the control subjects (88 +/- 5.5, P less than 0.005) or in 18 patients with hypothalamic or pituitary hypothyroidism (77.9 +/- 10.1, P less than 0.001). Following weight gain in 6 anorexia patients, there was a significant rise in serum T3 without change in T4Adj concentration. The Achilles reflex half-relaxation time (ART) in 38 anorexia patients was 348.6 +/- 10 msec compared with 280 +/- 30 msec in 168 normal age-matched subjects (P less than 0.001),  and was prolonged (greater than 340 msec) in 65% of these 38 patients. In 18 anorexia patients with measured ART, T3 and T4Adj, the mean ART was longer 376.1 +/- 20 msec) in 10 with subnormal T3 than in 8 patients with a normal T3 (294.7 +/- 13.2 msec, P less than 0.01). There was no significant difference in the mean ART between patients with a normal or low serum T4Adj. Administeration of oral T3 40 mug/day for 4 weeks to 11 anorexia patients caused a significant reduction (P less than 0.001) in mean ART of 108.7 +/- 9.6 msec compared with 17.7 +/- 3.3 msec in 18 normal subjects. There was a normal peak serum TSH and a rise in mean total serum T3 of 47 +/- 12 ng/100 ml (range 11-100 ng/ml) in 7 of 8 patients following 200 mug of iv thyrotropin releasing hormone (TRH). The fall in serum TSH was delayed in 6 patients. Assessment of hypothalamic control of thyroid function in 3 patients using the method of thyroidal iodide release (TIR) showed impairment of the normal diurnal variation and response to administered glucocorticoids. In the absence of a space-occupying pituitary lesion, the TRH and TIR data suggest a central inhibition of thyroid function, possibly by impairment of hypothalamic TRH release. In addition, a probable decrease of peripheral T4 to T3 conversion leads to low serum T3 concentrations. The prolonged basal ART and the marked ART reduction in response to T3 administration is attributed to correction of tissue thyroid hormone deficiency in the anorexia patients.


Failure of growth hormone-suppressing agents to affect TSH-releasing hormone- and LH-releasing hormone-induced growth hormone release in acromegaly.

In patients with acromegaly whose basal plasma GH levels were suppressed with 9 mg/day of dexamethasone for 2 days, TRH-(6 cases) and LHRH-(1 case) induced GH release were unaffected when the responses were compared to the basal levels. Phentolamine infusion, 70 mg in 150 min, or hyperglycemia induced by iv infusion of 700 ml of 50% glucose solution also did not suppress TRH-induced GH release in 2 acromegalic patients whose basal GH levels were lowered with these agents alone. These results seem to indicate that dexamethasone does not affect TRH- or LHRH-induced GH release per se, but affects the basal state which determines the absolute level of response. They also support the concept that TRH and LHRH act directly on pituitary tumor cells to release GH in acromegaly.


Effect of cyproheptadine on TRH-stimulated prolactin and TSH release in man.

TRH-stimulated prolactin release was determined in four male volunteers with and without pre-treatment with cyproheptadine. All showed a marked increase in the TRH-mediated prolactin rise and a marked decrease in the TRH-mediated TSH rise when receiving cyproheptadine pre-treatment.


Effect of triiodothyronine treatment on prolactin secretion in patients with amenorrhea-galactorrhea.

In order to determine the role of thyroid hormone in prolactin (PRL) secretion in patients with amenorrhea-galactorrhae, PRL response to 500 mug of iv thyrotropin-releasing hormone (TRH) was studied before and after the administration of triiodothyronine (T3) in 10 patients with amenorrhea-galactorrhea. Seven of these patients were euthyroid and the other 3 had hypothyroidism. The patients in the euthyroid group received 50 mug of T3 daily for 7 days and 75 mug q.d. for the ensuing 14 days. The hypothyroid patients received T3 at progressively increasing doses from 10 mug q.d. to 75 mug q.d. during 34 to 68 days. In the initial test, the elevated basal levels of PRL, 61.9 +/- 9.8 ng/ml (Mean +/- SE) exhibited a slight but insignificant net increase (7.7 +/- 2.1 ng/ml) after TRH injection in the euthyroid group. However, a marked response to TRH with a net increase of 147.2 +/- 26.3 ng/ml from the basal level of 47.3 +/- 11.2 ng/ml was observed in the hypothyroid patients. After treatment with T3, both the basal level (56.9 +/- 8.3 ng/ml) and the net increase (9.9 +/- 3.6 ng/ml) of PRL following TRH stimulation remained virtually unchanged in the euthyroid group. The hypothyroid group, in contrast, displayed a significant depression of both the basal level (26.1 +/- 13.0 ng/ml) and the net increase (33.8 +/- 6.5 ng/ml) of PRL to TRH stimulation. The diminution of the basal levels and responses of thyroid-stimulating hormone (TSH) to TRH stimulation was observed in all cases of both groups. These results suggest that the level of thyroid hormone has little pathogenic role in PRL secretion in euthyroid patients with amenorrhea-galactorrhea, in contrast to its marked effect in hypothyroid patients with amenorrhea-galactorrhea.


Studies on the nature of thyroidal suppression during acute falciparum malaria: integrity of pituitary response to TRH and alterations in serum T3 and reverse T3.

The nature of the suppression of the pituitary-thyroid axis during infection was studied by testing the integrity of thyrotropin (TSH) and prolactin (PRL) responses to thyrotropin-releasing hormone (TRH) during acute falciparum malaria in human volunteers. During infection, TSH responses to TRH were found to be intact while PRL secretion was slightly increased. That serum T3 levels abruptly declined during infection while serum T4 was stable or increasing suggested an alteration in peripheral degradative pathways and prompted the measuremnt of reverse T3. Changes in serum T3 concentration were found to be accompanied by reciprocal changes in reverse T3. These observations allow some clarification of previously unknown aspects of thyroidal economy during infection.


Diphosphonate therapy of paget's disease of bone.

The use of disodium ethane-1 hydroxy-1, 1-diphosphonate (EHDP) therapy for Paget's disease of bone was examined in 75 affected patients. Forty-eight patients received randomly assigned oral doses of either 0, 2.5, 5, 10, or 20 mg/kg/day in a controlled, double-blind protocol, and the remainder received either 10 or 20 mg/kg/day in a non-random protocol. The clinical status of the patients and appropriate laboratory tests were evaluated before treatment and at frequent intervals during a six-month period of initial therapy. There were no significant changes in either urinary hydroxyproline or serum alkaline phosphatase in those patients receiving placebos, while both these parameters decreased significantly at all dose levels of EHDP, with the greatest decline noted in the highest dose group. However, statistical analysis of the data related to changes in symptoms in the double-blind study revealed that patients receiving the higher dose of EHDP (10 or 20 mg/kg/day) had less favorable outcomes than those receiving the lower doses (2.5 or 5 mg/kg/day). The high does group had a relatively lower rate of symptom improvement and a relatively greater rate of deterioration than did the low dose group. Twenty-one of forty-nine patients followed for at least 18 months have shown a sustained suppression of their serum alkaline phosphatase and urinary hydroxyproline values for 12 months following cessation of EHDP, while therapy has been reinstituted for the other 28 patients because of increases in these measurements, with or without accompanying symptomatic deterioration. Eight patients sustained fractures through Pagetic bone during the period of study and all of these were treated with higher doses of EHDP. On the basis of the biochemical and clinical data in this study it appears that initial therapy of Paget's disease of bone with 5 mg EHDP/kg/day maximizes benefits while minimizing possible adverse effects.


Identification of Neisseria gonorrhoeae by genetic transformation: a clinical laboratory evaluation.

Transformation of a Neisseria gonorrhoeae auxotroph (uracil and arginine deficient) to prototrophy was attempted with wild-type deoxyribonucleic acid from 71 random clinical N. gonorrhoeae cultures. Of these 71 cultures, 97.1% transformed the nutritionally deficient mutant to prototrophy. The procedure was reliable and economical and offered several distinct advantages over other methods used for the confirmation of N. gonorrhoeae.


Asymptomatic females: detection of antibody activity to gonococcal pili antigen by radioimmunoassay.

A gonococcal pili antigen preparation was used to detect antibody activity sera obtained from 322 culture-positive asymptomatic females and 150 negative controls. Pili were obtained from a culture of type 2 Neisseria gonorrhoeae (strain 2686) and labeled with 125I for use in a double-antibody radioimmunoassay test system. Of the 322 sera obtained from culture-positive, asymptomatic females, 276 (85.7%) showed antibody activity greater than or equal to 1.8 mug/ml. Negative controls were obtained from three different groups of individuals, and 130 (86.7%) had undetectable antibody activity. Sera from asymptomatic, culture-positive females were absorbed with three different strains of N. gonorrhoeae, one of these strains being the organism used for pili antigen preparations. The absorbed sera were tested for antibody activity, and in each case the activity in the absorbed sera dropped to an undetectable level. When the same sera were absorbed with N. meningitidis, N. catarrhalis, N. perflava, Escherichia coli, Herellea vaginicola, Mima polymorpha, Staphylococcus aureus, and Candida albicans, little, if any, decline in the level of anti-pili antibody activity was observed.


Failure of cyclophosphamide to significantly enhance isolation of Rickettsia tsutsugamushi from wild Philippine rats.

Treatment of wild rats from a known scrub typhus focus in the Philippines with cyclophosphamide did not significantly enhance the isolation rate of Rickettsia tsutsugamushi using mouse inoculation techniques. Similarly, cyclophosphamide treatment of mice inoculated with organs of wild rats did not increase the recovery of rickettsiae, and isolation of rickettsiae was about equal from biopsies obtained before and after immunosuppression of wild rats.


Effect of N-acetylcysteine on antibiotic activity and bacterial growth in vitro.

The antibiotic bacerial inactivity of N-acetylcysteine (NAC) and its interaction with penicillin and aminocyclitol antibiotics was evaluated. NAC inhibited growth of both gram-negative and gram-positive bacteria. Strains of Pseudomonas aeruginosa were more susceptible than other microorgainsms tested. P. aeruginosa strains were inhibited synergistically by NAC and carbenicillin or ticarcillin. However, NAC antagonized the activity of gentamicin and tobramycin. These findings have implications for the combined clinical use of NAC and aerosolized antibiotics and are also important for the processing of sputum specimens in the microbiology laboratory.


Simplified complete medium for the growth of Neisseria gonorrhoeae.

A complete medium for the growth of Neisseria gonorrhoeae has been developed, using the same basic ingredients contained in commercial media but with fewer supplements. Based on a comparison of plating efficiencies, this medium (designated TTU Complete Medium) supported the growth of laboratory strains of N. gonorrhoeae equal to that obtained on commercial GC agar base medium supplemented with Iso VitaleX. It was also equivalent to the commercial medium in supporting the growth of 67 clinical isolates and 6 auxotypes of N. gonorrhoeae when tested by the streak plate method. Based on turbidity measurements and viable plate counts, the liquid equivalent of TTU Complete Medium supported maximal growth yields of N. gonorrhoeae. The toxicity of different brands of agar appeared to be a major factor in preventing the growth of N. gonorrhoeae strains on solid media. The addition of starch neutralized the toxicity of some types of agar but not of others.


Comparative serological and cutaneous reactivity of candidal cytoplasmic proteins and mannan separated by affinity for concanavalin A.

Yeast-form Candida albicans cells were disrupted for 1.5 min in a Braun homogenizer and centrifuged at 100,000 X g. The supernatant was concentrated by ammonium sulfate precipitation and then dialyzed. The resulting material (650 mg), containing 81.2% protein and 11.5% carbohydrate, was subjected to affinity chromatography on concanavalin A (Con A) linked to agarose. A protein fraction was eluted from the column with buffer, and a fraction containing mannan was eluted with 0.2 M alpha-methyl mannoside. The candidal soluble proteins had 19 components which were resolvable by polyacrylamide gel electrophoresis. The material with affinity for Con A contained mannan and 17% complexed protein. Antigenic differences between the soluble proteins and the mannan-protein complex were shown by lines of intersection in immunodiffusion. The soluble proteins devoid of mannan reacted in immunoelectrophoresis with sera from infected rabbits and patients with chronic candidiasis. These same sera also reacted with a mannan-protein complex eluted from the Con A column with alpha-methyl mannoside. The comparative ability of candidal proteins and cell wall-derived mannan to elicit skin test reactions in guinea pigs sensitized by infection or with formaldehyde-killed yeast was studied. Candidal proteins at a 10-mug dose elicited positive reactions at 6 and 21 days after sensitization. The reactions persisted for 48 h and showed minimal tendency to an arthus response, which was marked when mannan-containing antigens were used. The antigenicity of cell wall-derived mannans and candidal soluble proteins devoid of mannan was compared in immunodiffusion tests of sera from 39 patients with neoplastic disease. Of these patients with documented candidiasis, 13 of 20 reacted to one or more mannan antigens, and 3 of 20 reacted to candidal soluble proteins. In contrast, of those patients who were uninfected or had superficial Candida spp. infections, 5 of 19 reacted to candidal soluble proteins, and 16 of 19 reacted to one or more mannan antigens.


Mechanism for the inflammatory response in primate lungs. Demonstration and partial characterization of an alveolar macrophage-derived chemotactic factor with preferential activity for polymorphonuclear leukocytes.

Approximately 4 h after an initial bronchoalveolar lavage (BAL) of a primate's lung, an appreciable number of polymorphonuclear leukocytes (PMNs) were noted to accumulate in respiratory fluids when lavage was repeated. Whereas, alveolar macrophages (90%) and lymphocytes (7%) were the principal respiratory cells recovered initially from lavage fluid, later samples contained 45-90% PMNs To explain the observed ingress of PMNs into lung fluids, concentrated BAL fluid was tested for chemoattractant activity. Such fluid obtained 4 and 24 h after an initial lavage contained material that produced directed migration (chemotaxis) for PMNs and mononuclear cells isolated from peripheral blood of normal donors. Gel filtration chromatography of BAL disclosed two peaks of chemotactic activity in the effluent fractions. Material from the column with an estimated molecular weight of 15,000 daltons was chemotactic for both PMNs and mononuclear cells. Because it was susceptible to inactivation with antiserum against the fifth component of complement, resistant to heating, and unaffected by antiserum against C3, this factor was considered analogous to the cleavage product of the fifth component of complement. C5a. In addition chemotactic activity for PMNs only was contained in an effluent peak having a molecular weight of about 5,000 daltons. This material was heat labile but unaffected by antisera to complement components. To locate the possible source of these factors in respiratory fluid, in vitro cultures of alveolar macrophages were established. These cells, whether stimulated by phagocytosis of opsonized bacteria or merely by attachment to a glass surface, produced chemotactic material which had physical characteristics similar to the small molecular weight material in BAL. Moreover, it induced preferential chemotaxis for PMNs. Thus, in primate lungs, at least two chemotactic substances may generate an inflammatory response; one which is a fragment of the complement component C5 and another small molecular weight factor which is released from alveolar macrophages.


Age-related deterioration of pyramidal cell basal dendrites in rat auditory cortex.

The basal dendritic trees of layer V pyramidal cells in the rat auditory cortex were examined quantitatively in a group of 3-month-old and a group of 34- and 36-month-old rats. Two forms of analysis were used on the Golgi preparations: (1) the number of intersections between the basal dendrites and a series of concentric circles whose common center lies over the perikaryon center, and (2) the number of dendritic branches, by order, per neuron. The data indicate that in the old animals the density of the dendritic tree has decreased significantly within a radius of about 150mu of the perikaryon, yet the extent of the dendritic domain has not changed appreciably. Analysis of the dendritic branching suggests that there has been a deterioration not only in the peripheral branches of the dendritic tree, but also that entire dendrites have been lost. This loss of primary branches was confirmed through the reconstruction of layer V neuronal perikarya and their proximal dendrites from 1-mu plastic serial sections of auditory cortex. Concomitant with the loss of dendrites which accompanies advancing age is a tendency for the perikaryon to be smaller, but not distorted, in the old animals.


Demonstration of bilateral projection of the central retina of the monkey with horseradish peroxidase neuronography.

In the primate, ganglion cells of the temporal retina project ipsilaterally and those of the nasal retina, contralaterally into the optic tract. The vertical meridian passing through the fovea defines the border between these two populations of ganglion cells and has been demonstrated in four Macaque monkeys after unilateral injection of horseradish peroxidase into the dorsal lateral geniculate nucleus and examination of the pattern of retrograde labeling of those ganglion cells projecting to the injected side. A median 1 degree vertical strip in which ipsi- and contralaterally projecting ganglion cells intermingle was found, confirming the report by Stone et al. ('73). In addition, occasional extrafoveal labeled ganglion cells were found as far as 2 degrees from the vertical midline in the otherwise unlabeled hemiretinae. These ganglion cells were not numerous and had somata of all sizes, suggesting that they do not constitute a separate class of ganglion cells as found in the temporal retina of the cat. In contrast to the description by Stone et al. ('73), the strip of vertical overlap did not show a constant width through the fovea, since mixing of labeled and unlabeled ganglion cells was found in a band approximately 1/2 degree wide along both the nasal and temporal rims of the foveal pit which is 500 mum (2 degrees) in diameter. Beyond these 1/2 degree arcs, the appropriate hemiretina was either completely unlabeled, or contained virtually every ganglion cell labeled on the side projecting to the injected dorsal lateral geniculate nucleus. The scattered labeled ganglion cells rimming an otherwise unlabeled hemifovea represent a possible anatomical basis for the phenomenon of "macular" or "foveal sparing" in which unilateral damage to the occipital cortex produces homonymous hemianopsia with sparing of a small island of centralmost vision extending about 1 degree from the foveal center. From this study, it is not possible to define the receptive fields or specific photoreceptor connections of the ganglion cells labeled with horseradish peroxidase, so that at the present time quantitative correlations cannot be made between the numbers of ganglion cells remaining on the affected side of the fovea and the extent of preservation of visual function in the spared zone. The presence of labeled ganglion cells rimming the fovea in its entirety is compatible with the sequence of foveal development in late prenatal life. After lateral displacement both nasally and temporally of ganglion cells which initially lay in the median vertical overlap strip of 1 degree, in the adult retina a strip approximately 1/2 degree wide around the perimeter of the foveola should contain a mixture of ipsi- and contralaterally projecting ganglion cells. The total population of ganglion cells beyond this 1/2 degree band should be completely ipsi- or contralateral in their projection patterns, as is observed...


Thalamic projections of the superior colliculus in the rhesus monkey, Macaca mulatta. A light and electron microscopic study.

The projections of the superior colliculus to the thalamus have been studied in the monkey, Macaca mulatta, with anterograde degeneration techniques. The superior colliculus has been shown to project to the inferior nucleus of the pulvinar in a topographical manner with the lower visual field represented dorsomedially and the upper field ventrolaterally. The peripheral zone is located along the medial border and the fovea at the dorsolateral angle adjacent to the lateral geniculate nucleus. The superior colliculus also sends a dense projection to the ipsilateral intralaminar complex, i.e., to the parafascicular, central lateral and paracentral nuclei, and a lesser projection to the same contralateral nuclei. Degenerating tectal fibers were also found in the lateral geniculate nuclei. Four types of vesicle containing profiles were observed in the inferior pulvinar and paracentral nucleus. The large RL and small RS terminals contain round vesicles of uniform size and form asymmetric contacts mainly with large and small dendrites respectively. The F terminal contains a mixture of small round and flat vesicles. It forms symmetric contacts with dendrites and cell somata. The P profile is very pale and contains a relatively sparse population of vesicles showing a great variation in size. It forms symmetric contacts with medium to large dendrites. It is frequently found postsynaptic to the other types, especially the RL terminal, and is regularly seen as the intermediate element of serial and triadic synaptic arrangements. The experimental electron microscopic study has shown that many fibers from the superior colliculus terminate as RL profiles, undergoing direct dense degeneration, in both the inferior pulvinar and the paracentral nucleus. Others probably end as smaller RS terminals.


An intricately patterned prefronto-caudate projection in the rhesus monkey.

The distribution of prefronto-caudate fibers in the caudate nucleus was studied autoradiographically in monkeys of various ages in which tritiated amino acids had been injected into the middle one-third of the length of the dorsal bank of the principal sulcus. The results indicate that, contrary to previous reports which had suggested a projection to only the head of the caudate nucleus, area 9 of Brodmann projects to the entire length of the nucleus. In the head of the caudate nucleus the cortico-caudate fibers are distributed in a pattern which is remarkable in two respects. First, the grains are not uniformly distributed but rather are segregated into clusters separated from one another by territories in which grain density does not exceed background. Second, individual clusters of grains, circular or elliptical in shape, surround grain free cores. These patterns of fiber distribution within the head of the nucleus are more sharply defined in newborn than in older monkeys. Our findings suggest that the caudate nucleus is organized more as an anatomic and functional mosaic than as the homogeneously organized structure that it is commonly considered to be.


Cortical projections of posterior parietal cortex in owl monkeys.

Efferent cortical projections of posterior parietal cortex were determined by degeneration and autoradiographic methods in owl monkeys. Intraregional connections were to the immediate surround of the injection or lesion site, and to distinct foci within the posterior parietal region. The extraregional ipsilateral connections were with (1) previously established subdivisions of visual association cortex (the Dorsomedial Area, the Medial Area, the Dorsolateral Area, and the Middle Temporal Area), (2) other locations in caudal neocortex, and (3) frontal cortex. The callosal projections were to separate foci in posterior parietal cortex of the contralateral cerebral hemisphere. The separate foci of both ipsilateral and contralateral terminations in posterior parietal cortex raise the possibility that this region contains more than one functional subdivision. The connections with visual association cortex suggest a role for parietal cortex in visual behavior. Other foci in caudal neocortex indicate the possible locations of additional subdivisions of association cortex.


Allergens in bee venom II. Two new high molecular weight allergenic specificities.

Two new allergenic specificites were detected in honeybee venom and the two corresponding protein substances isolated by gel filtration, immunoadsorption, and ion exchange chromatography. The first of these, allergen B, has a molecular weight ranging from 49,000 to more than 200,00 d and can be recognized by rabbit and guinea pig antisera as well as by human reaginic sera using the radioallergosorbent test (RAST). Allergen B gives a single line in immunodiffusion distinct from hyaluronidase, phospholipase A, melittin, and the other high molecular weight substances described and gives a single band at 49,000 d in sodium dodecyl sulfate (SDS) polyacrylamide gel. The second substance, allergen C, has a molecular weight of 105,000 d and was separated from allergen B by immunoadsorption with insoluble antibody. Allergen C was shown to be distinct from the other sustances in bee venom by immunodiffusion with animal antisera. One human reaginic serum was monospecific for allergen C. Two other minor components of 86,000 and 71,000 d are present in bee venom; their allergenic activities are unknown. The two specifities, B and C, comprise most of the reactivity of the previously described Sephadex G-75 fraction 1 and clearly are important allergens, reacting with 98% of sera from bee venom-allergic individuals.


Aldose reductase and sorbitol dehydrogenase distribution in rat kidney.

The sorbitol pathway catalyzes the conversion of glucose to fructose via the intermediate sorbitol. It consists of aldose reductase (AR) and sorbitol dehydrogenase (SDH). In adult (44 day) kidney zones, AR was highest in the outer medulla. In substructures AR was highest in distal convoluted tubule. The AR was greatest in newborn and 8-day zones of developing rat kidney. Acute alloxan diabetes was associated with decreased AR in small arteries, but not glomeruli. The SDH was lowest in outer medulla. It was most active in glomeruli and distal convoluted tubules. The diabetic state leads to no change of SDH in arteries but an increase in glomeruli. SDH increased with development. This study demonstrates AR and SDH in substructures of the kidney. The pathway is present in developing kidney. In diabetes the enzymatic changes would tend to decrease accumulation of sorbitol.


Serotypes of Bacillus cereus from outbreaks of food poisoning and from routine foods.

A provisional serotyping scheme was used to type cultures of Bacillus cereus from 84 outbreaks of food poisoning in seven countries; 283 of the 337 (84%) cultures tested were typable. In 35 of the 61 outbreaks associated with a vomiting-type syndrome, foods, clinical specimens or both yielded H-serotype 1 only. Type 1 strains together with other sterotypes were isolated in seven outbreaks. In 14 outbreaks types 3, 4, 5, 8 or a mixture of serotypes were present. Untypable strains were isolated in five outbreaks. Two of the nine diarrhoeal-type outbreaks yielded serotype 1 only. Types 2, 6, 8, 9, 10 and a mixture of type 12 and an untypable strain appeared to be responsible for one outbreak each. Although 16 of the 18 recognized serotypes were present among cultures of B. cereus from various routine foods, only 156 of the 400 (39%) isolates tested were typable.


Immunogenetic studies of rhesus monkeys. VII. A simple hemagglutination technique for blood typing.

A microtiter hemagglutination technique was adapted for routine blood typing of rhesus monkeys. The main advantage of the technique is economy of scarce reagents, since they can be used at greater dilutions and in smaller quantities than in the standard tube technique. Furthermore, the technique is sensitive, accurate, and easy to perform.


Wheat grains: a substrate for the determination of gluten antibodies in serum of gluten-sensitive patients.

A simple method of detecting gluten antibodies in serum is described. Cryostat sections of wheat grains proved to be an excellent substrate in the immunofluorescence technique. Rabbit antisera to gliadin and alpha-gliadin, and high percentage of sera from patients with gluten-induced enteropathy had antibodies that reacted with an internal structure of wheat grains. These antibodies could be absorbed with gliadin and with alpha-gliadin.


Granule-bound tyrosinase: solubilization and its relation to the soluble form of tyrosinase.

Disc gel electrophoresis of tyrosinase solubilized by sodium deoxycholate (DOC) from the particle fraction of B16 mouse melanoma was carried out in the presence of DOC. A single tyrosinase band, considered to be T3, was detected at the Rx value of 0.60 against Coomassie brilliant blue. The T3 band which is located between T1 and T2 seems to be distinct from the soluble forms of tyrosinase. Its molecular weight was estimated to be 102,000. When treated with proteolytic enzymes or refrigerated, the T3 molecule converted into a molecule whose mobility was equivalent tothat of T1.


Detection by immune electron microscopy of 26- to 27-nm viruslike particles associated with two family outbreaks of gastroenteritis.

Viruslike particles 26-27 nm in size were detected by immune electron microscopy in stools of volunteers who were ill after administration of bacteria-free fecal filtrates derived from two separate family outbreaks of acute epidemic nonbacterial gastroenteritis. Fluorocarbon treatment and concentration of the filtrates were necessary to provide enough antigen to test sera by immune electron microscopy. Serum antibody responses were detected in both naturally occurring and experimentally induced cases of illness. The Montgomery County viruslike particle appeared to be related to the previously described Norwalk particle, whereas the Hawaii particle appeared to be unrelated to the Norwalk particle.


Synergism of penicillin and gentamicin against Listeria monocytogenes in ex vivo hemodialysis culture.

A new technique of ex vivo hemodialysis culture was used to study the effects against Listeria monocytogenes of penicillin G and gentamicin administered intra-muscular, singly and in combination, in the goat. Bactericidal synergism of the combination was detected despite rapidly diminishing concentrations of the drugs in the blood plasma and dialysate.


Comparison of amikacin and tobramycin in the treatment of infection in patients with cancer.

Two new aminoglycoside antibiotics, tobramycin and amikacin, were compared in a randomized study of the treatment of infections in patients with cancer. For the identified infections, the response rate for tobramycin was 60% and for amikacin was 64%. Pneumonia, urinary tract infection, and septicemia were the most frequent infections. Most (78%) of the identified pathogens were gram-negative bacilli; Klebsiella pneumoniae, Escherichia coli, and Pseudomonas aeruginosa were the most frequently isolated organisms. When only infections due to gram-negative bacilli were considered, 67% responded to tobramycin and 69% responded to amikacin. All infections except pneumonias had at least a 50% response rate to either antibiotic. The major form of toxicity of both antibiotics was azotemia and occurred in 22% of cases treated with tobramycin and in 20% treated with amikacin. Tobramycin and amikacin are equally effective in the treatment of gram-negative infections and have similar toxicity.


Spleen function in quartan malaria (due to Plasmodium inui): evidence for both protective and suppressive roles in host defense.

For investigation of the role of the spleen in host defense and chronicity in quartan malaria, the course of Plasmodium inui infection was studied in 39 intact rhesus monkeys, 16 monkeys splenectomized before infection, and 22 monkeys splenectomized after infection. Splenectomized animals consistently had peak parasitemias greater than 10-fold higher than those of intact animals, but the mortality rate at peak parasitemia was only higher in monkeys splenectomized before infection. Whereas intact animals had persistent infections for one to 13 years, splenectomized animals achieved self-cure (persistently negative blood smears for three or more months) within less than a year. These observations suggest that the spleen exerts a protective role during the acute stage and a suppressive role during the chronic stage of quartan malarial infection.


Extraction and purification of specific antibody against Pseudomonas aeruginosa by immune affinity chromatography.

Immune affinity chromatography has been used to extract and purify specific immune antibody against P. aeruginosa immunotype 4 from hyperimmune gamma glubulin containing antibody against each of seven major immunotypes of P. aeruginosa. A column with a 4 ml. bed volume was construced which had a "working capacity" of approximately 1,680 agglutination units. With water used as the eluting agent, 100 percent yields of functionally active antibody were obtained when amounts equal to or less than the working capacity were loaded onto the column. The technique appears to be suitable for large scale extraction of specific immune antibody for therapeutic trials or investigative use.


Knee effusions.

There are many questions concerning knee effusions and the basic diagnostic approach to them. The case of a young female with gonococcal arthritis presenting to the Emergency Room with an isolated knee effusion is discussed. From the errors in diagnosis of this case, a basic outline in the approach and work-up of a knee effusion is detailed, including history and physical examination, and the use of synovial fluid analysis as a further diagnostic aid. Procedural instructions and contraindications for knee arthrocentesis are given, as well as data related to interpretation of pertinent laboratory findings of synovial fluid in disease states.


T-cell-mediated concomitant immunity to syngeneic tumors. I. Activated macrophages as the expressors of nonspecific immunity to unrelated tumors and bacterial parasites.

Progressive growth of the SA1 sarcoma was shown to result in the generation of a state of concomitant resistance to growth of a second implant of the same tumor. The responding lymph nodes of concomitantly immune mice were shown to contain theta-positive T cells that could specifically neutralize the growth of tumor cells in a normal test recipient. Nevertheless, the concomitantly immune host itself was capable to a limited extent of suppressing the growth of unrelated tumors. The generation of immunity, moreover, was associated with the generation of a powerful state of macrophage-mediated, nonspecific resistance to the bacterial parasite, Listeria monocytogenes. It was concluded that systemic macrophage activation was the consequence of the generation of T-cell-mediated immunity to the progressively growing tumor, and that this not only gave the host the capacity to inhibit the growth of unrelated tumors, but also to protect itself against microbial infection. The results gives credence to the view that macrophages play a central role in defense against microbial and neoplastic growth.


Influence of magnesium on chloride-induced calcium release in skinned muscle fibers.

Chloride-induced Ca release in skinned muscle fibers was studied by measuring isometric force transients and 45Ca loss from fiber to washout solutions. Skinned fibers prepared from muscles soaked in normal Ringer solution made large force transients in 120 mM Cl solution with 5 mM ATP and 1 mM Mg, but 3 mM Mg was inhibitory. Mg inhibition was antagonized by low temperature and by Cd, agents which slow active Ca uptake by the sarcoplasmic reticulum (SR). In low Mg++, Cl stimulated rapid 45Ca release from the SR in sufficient amounts to account for the force response. The increased 45Ca release was inhibited by EGTA, suggesting that release requires free Ca under these conditions. The 45Ca initially released was partially reaccumulated later. Reaccumulation was increased in higher Mg++. These results provide additional evidence that the Ca uptake rate is an important determinant of net release, and suggest that Mg++ acts primarily on this mechanism. Skinned fibers prepared from muscles soaked in low Cl solutions could give force responses to Cl solutions with 3 mM and 6 mM Mg. This observation suggests that the Cl stimulus varies with the [Cl] gradient across the internal membranes, and supports the hypothesis that applied Cl causes membrane depolarization.


Solid media containing carboxymethylcellulose to detect CX cellulose activity of micro-organisms.

Solid media containing carboxymethyl cellulose (CMC) were developed to detect CX cellulose-producing micro-organisms. Hydrolysis of CMC was seen as a clear zone around colonies after flooding plates with 1% aqueous hexadecyltrimethyl-ammonium bromide. Tests with ten bacterial and four fungal species showed that the degree of substitution (DS) of the CMC affects both growth and enzyme production. Most of the organisms produced more CX cellulase on CMC with a DS of 0-9, but CMC with a DS of 0-4 was better for one fungus. A qualitative measure of cellulase production may be obtained by calculating the ratio of zone size to colony diameter. Solid media containing CMC provided a more rapid assay of CX cellulose production than a medium containing native cellulose.


Construction and properties of hybrids obtained in interspecific crosses between Streptomyces coelicolor A3(2) and Streptomyces griseus Kr-15.

Recombinants between Streptomyces coelicolor A3(2) and Streptomyces griseus Kr-15 were obtained using methods of hybrid construction. Recombinant Rcg1, obtained from a cross between S. griseus and a S. coelicolor UF (SCPI-) strain, phenotypically resembled S. coelicolor UF strains and in crosses with a S. coelicolor NF donor strin produced recombinatn progeny at a frequency of 100%. Recominant Rcg3, like SCP1-carrying S. coelicolor strains, inhibited SCP1-strains of S. coelicolor and in crosses with a UF recipient strain of S. coelicolor generated recombinants at high frequency. In crosses between S. griseus and Rcgi the frequency of recombinant formation was increased about 100-fold relative to crosses between S. griseus and S. coelicolor. Effective transfer of S. grieseus and Rcg3 chromosomal markers into Rcg1 and S. coelicolor, respectively, indicated that S. griseus had donor properties. Studies of the ability of recombinants to support phage growth indicated that parental chromosomal fragments containing genes involved in control of phage-receptor formation and intracellular growth were present in the hybrids. Grisin-producing recombinants, capable of restricting phages attacking S. coelicolor and S. griseus, were obtained.


Fatty and mycolic acid composition of Bacterionema matruchotii and related organisms.

Whole-organism methoanolysates of bacterionemae contained mycolic acids in addition to other long-chain fatty acids. These mycolic acids were similar in general structure and overall size to those found in strains of Corynebacterium diphtheriae and Corynebacterium xerosis. The long-chain fatty acids of bacterionemae, mainly straight-chain saturated and unsaturated acids, were similar to those of certain coryneform bacteria including C. diphtheriae. On the basis of these lipid data, and results of earlier studies, we recommend that the genus Bacterionema be transferred from the family Actinomycetaceae to the Coryneform Group of Bacteria.


Genetic transformation in Methylobacterium organophilum.

Several mutants have been isolated from the facultative methylotroph, Methylobacterium organophilum, using either N-methyl-N'-nitro-N-nitrosoguanidine or ultraviolet light as mutagens. One of these isolates, a glutamate auxotroph lacking isocitrate dehydrogenase, has been transformed to prototrophy, using wild-type DNA, at a frequency of 0-5%. Competence and DNA uptake occur only in cultures which are near the end of exponential growth, and maximal transformation requires a DNA concentration of 100 mug ml-1.


The metabolism of starch, glucose, amino acids, purines, pyrimidines and bacteria by the rumen ciliate Polyplastron multivesiculatum.

The large rumen ciliate protozoon Polyplastron multivesiculatum grown in vitro engulfed a wide range of bacteria (from a population density of 10(9) bacteria ml(-1)) at a rate of 1500 to 137000 bacteria h(-1) protozoon(-1). No evidence was found for the preferential engulfment of bacteria of rumen origin. Except for Proteus mirabilis none of the bacteria were digested with the liberation of soluble materials into the medium. Glucose and amino acids were taken up rapidly by P. multivesiculatum compared with the rate of uptake by Entodinium caudatam. Glucose was incorporated into protozoal polysaccharide and into bacteria associated with the protozoa and was used for the synthesis of a wide range of amino acids. Evidence showed that bacteria and free amino acids at the concentrations found in the rumen could supply the protein requirements of the protozoa for division at least once each day.


Reversibility of the adsorption of bacteriophage PL-1 to the cell walls isolated from Lactobacillus casei.

Bacteriophage PL-1 adsorbed specifically to fragments of the isolated cell walls of its host Lactobacillus casei ATCC 27092 and failed to adsorb to cell wall fragments of resistant strains. Soon after mixing, an equilibrium situation of phage adsorption was attained. The equilibrium position was dependent on the cell wall concentration, but was not affected by the incubation temperature. The adsorbed phages were not inactivated by the cell wall fragments, but formed phage-cell wall complexes maintaining original phage infectivity. The infectivity of phage-cell wall complexes was neutralized by antiphage sera in the same manner as free phages. When the phage-cell wall complexes were repeatedly washed by centrifuging and resuspending in a fresh medium, the adsorbed phages were eluted as infective virions, confirming that phage adsorption was reversible. When the reactants concerned were allowed to approach equilibrium from the opposite direction, the same equilibrium state was achieved. The value of the euqilibrium constant (Keq) with respect to reversible adsorption was constant with various phage concentrations under the conditions used here. When a mixture of phages and cell walls at an equilibrium state was diluted, the unadsorbed phages increased in accordance with the decrease in the concentration of the reactants.


Further observations on subacute necrotizing encephalomyelopathy in adults.

The clinical and neuropathlogical aspects of four cases of subacute necrotizing encephalomyelopathy (SNEM) in adults, are described. In one, the disease was precipitated by a surgical procedure, and caused death; this resembled cases previously reported. In the other three, the disease was recognized as an incidental pathological finding, without apparent morbidity. In these, the pathologic lesions were few in number, but were characteristic in location and nature, most specifically in the preservation of neurons within the devasted tissues. The disease in adults is thought due to a less severe metabolic defect than that which causes death in infants, and may be more common. When extrinsic stress is superimposed, the disease may become severe and cause death as in the first case and those previously reported. This might be prevented by the prophylactic use of those therapeutic agents which have had a favorable effect in infants, particularly very high doses of thiamine. The prevelence of the disease in adults, including the incomplete cases included in this report, suggests that such prophylactic measures may be warranted when stress is anticipated, as with surgery.


Response of primate joint afferent neurons to mechanical stimulation of knee joint.

1. One hundred thirty-eight knee joint afferents from posterior articular nerve (PAN), in primates, were recorded in dorsal root filaments. Responses of afferents were studied in relation to both passive manipulations of the knee and active contractions of quadriceps, semimembranosus, and gastrocnemius muscles. 2. When the knee was passively rotated, most neurons discharged only when extreme angular displacements were achieved. Response of neurons responding to passive extensions was linearly related to the torque applied to the knee. With maintained extensions, discharge in extension neurons adapted slowly. Some of the time constants of adaptation were similar to those for simultaneously recorded torque relaxation. 3. Contractions of quadriceps, semimembranosus, or gastrocnemius muscles could activate many neurons in the absence of changes in joint angle. For quadriceps-activated neurons, rather high torques (mean = 2,450 g with cm) were required. 4. The results support the hypothesis that joint afferents function as capsullar stretch receptors, responding to those mechanical events which result in loading of the capsule.


Role of striate cortex and superior colliculus in visual guidance of saccadic eye movements in monkeys.

1. We studied the effect of lesions placed in striate cortex or superior colliculus on the detection of visual stimuli and the accuracy of saccadic eye movements. The monkeys (Macaca mulatta) first learned to respond to a 0.25 degrees spot of light flashed for 150-200 ms in one part of the visual field while they were fixating in order to determine if they could detect the light. The monkeys also learned in a different task to make a saccade to the spot of light when the fixation point went out, and the accuracy of the saccades was measured. 2. Following a unilateral partial ablation of the striate cortex in two monkeys they could not detect the spot of light in the resulting scotoma or saccade to it. The deficit was only relative; if we increased the brightness of the stimulus from the usual 11 cd/m2 to 1,700 cd/m2 against a background of 1 cd/m2 the monkeys were able to detect and to make a saccade to the spot of light. 3. Following about 1 mo of practice on the detection and saccade tasks, the monkeys recovered the ability to detect the spots of light and to make saccades to them without gross errors (saccades made beyond an area of +/-3 average standard deviations). Lowering the stimulus intensity reinstated both the detection and saccadic errors...


Brain-tumor chemotherapy. Pharmacological principles derived from a monkey brain-tumor model.

An implnated choriocarcinoma growing inthe brains of monkeys was used as a brain-tumor model for the study of the cerebral distribution of two commonly used physiological markers, inulin and albumin; tissue samples were obtained from the tumor, adjacent brain, and distant brain. An extravascular inulin space was calculated by subtracting the albumin (plasma) space from the total inulin spaces. The extravascular inulin space in the tumor was found to be 24%, a value significantly larger than that in distant brain (0.6%). The large inulin space of the tumor was probably the result of increases on both capillary permeability and the extracellular space within this area. Determination of the inulin space in 1 to 2-mm thick samples of tissue taken serially from the tumor center to the distant brain indicated a gradual decline in inulin concentration from the tumor's edge to distant brain. This distribution pattern could be the result of either a continuous decrease, running from the tumor to distant brain, in capillary permeability to inulin, or a diffusional flow of inulin from the tumor into the adjacent tissue. The failure of drugs to inhibit such a tumor in view of these observations is discussed.


A multidetector scintillation camera with 254 channels.

A computer-based scintillation camera has been designed for both dynamic and static radionuclide studies. The detecting head has 254 independent sodium iodide crystals, each with a photomultiplier and amplifier. In dynamic measurements simultaneous events can be recorded, and 1 million total counts per second can be accommodated with less than 0.5% loss in any one channel. This corresponds to a calculated deadtime of 5 nsec. The multidetector camera is being used for 133Xe dynamic studies of regional cerebral blood flow in man and for 99mTc and 197 Hg static imaging of the brain.


Thyroid function in prenatally protein-deprived rats.

Thyroid function was examined in fetal and newborn young of dams fed diets containing 24% or 4% casein as the sole source of protein. Radioactive iodine uptake, as examined autoradiographically and by gamma counting, was significantly reduced in glands of progeny of deficient rats. Following injections of thyrotropin or thyrotropin releasing hormone, thyroid cells of newborn young of protein-deprived rats contained fewer colloid droplets than those of control pups. Circulating levels of triiodothyronine and thyroxine were significantly reduced in the plasma of newborn young of protein-deprived females as compared with controls.


Amino acid fortification of bread fed at varying levels during gestation and lactation in rats.

The effects of protein quality improvement of white bread fed to female rats during pregnancy and lactation, under varying degrees of adequacy in food energy, on growth and brain development of the offspring have been studied. Weaning weight and the weight, protein and DNA of whole brain and major regions of the brain at weaning were determined. Lysine addition significantly increased all parameters, and threonine addition gave additional significant increments. Restricting energy intake decreased most parameters independent of protein quality. Brain cellularity was significantly higher in offspring of dams fed lysine and threonine fortified bread than in offspring of dams fed casein. All parameters measured in the offspring of dams fed lysine and threonine fortified bread at 70% ad libitum consumption were significantly increased over the values obtained in the offspring of dams fed unfortified bread ad libitum even though they had consumed 13% less protein  and dietary energy during the pregnancy and lactation period. The implications of these findings for practical problems in human nutrition are considered.


Blood flow changes after sagittal split of the mandibular ramus.

Blood flows to various cephalic tissues were determined by the microsphere method before and after unilateral sagittal split of the mandible in macaque monkeys. Significant blood flow decreases were found in certain bony regions of the osteotomized mandible while blood flow to the masseter and medial pterygoid muscles on theosteotomized side was significantly increased. Judicious stripping of the mucoperiosteum and pterygomassteric sling may be an important consideration when planning the sagittal split approach to correct certain mandibular anomalies.


Nonoperative treatment of duodenal hematomata in childhood.

Blunt abdominal trauma, often apparently trivial in nature, may produce intramural hematoma causing complete duodenal obstruction. A child with abdominal pain, persistent vomiting and upper abdominal tenderness should be carefully questioned for a history of trauma. Prompt upper gastrointestinal x-rays will allow early and definitive diagnosis. Previously recommended treatment varies from immediate surgical intervention to observation followed by exploratory laparotomy at a later date. We have treated nine patients with duodenal obstruction caused by intramural hematoma of which eight were successfully managed with nasogastric suction and parenteral fluid administration. One patient had the hematoma evacuated at surgery. This report reviews our experience.


Biological effects of cocaine derivatives I: Improved synthesis and pharmacological evaluation of norcocaine.

An improved synthesis of norcocaine, a metabolite of cocaine, is reported. Following intravenous administration to a rhesus monkey, respiratory effects were similar to those observed following cocaine treatment. In addition, clonic convulsions, hypothermia, and mydriasis resulted. Norcocaine could be responsible for part of the pharmacological activity of cocaine.


Sodium cyanate-induced ocular lesions in the beagle.

Ocular lesions secondary to sodium cyanate administration in the beagle are described. Fifteen dogs were given sodium cyanate at doses of 30 to 170 mg/kg/day, 5 days per week. Fourteen developed cataracts which were generally posterior and subcapsular. Five animals developed corneal lesions. Control beagles and cyanate-treated rats and monkeys did not develop lesions. The one drug-treated beagle without ocular lesions received a maximum dose of 30 mg/kg/day of sodium cyanate for 19 months and achieved a mean carbamylatin of 0.39 residue of cyanate per mol of hemoglobin. Animals developing cataracts received a maximum dose of 119+/-23 mg/kg/day for 28+/-10 months and achieved a mean carbamylation of 0.69+/-0.13 residue/mol. Animals developing corneal lesions received a maximum dose of 113+/-22 mg/kg/day for 36+/-7 months and achieved a mean carbamylation of 0.78+/-0.09 residue/mol. Younger animals appeared more susceptible to the cataractogenic effects of the drug than older ones. Lens intracellular sodium was increased by 100% in the cataractous lenses while potassium was significantly decreased in the lenses with the greatest opacity. Extracellular space was increased in the cataractous lenses by 61%. Reduced glutathione levels were decreased by 34% and lactate leakage into the medium was increased by 12%. Rubidium-86 uptake was increased by 26% in the cataractous lenses, probably reflecting an inadequate compensatory response to increased passive fluxes of cations resulting from a structural or enzymatic lesion.


Antiarrhythmic properties of MJ 9067 in acute animal models.

A novel benzanilide derivative, MJ 9067, has been shown to abolish experimental atrial and ventricular arrhythmiase effectively and promote the return of normal sinus rhythm in a variety of animal models. At intravenous dose levels ranging from 0.5 to 3.2 mg/kg, MJ 9067 successfully converted atrial fibrillation induced by either local application of aconitine or electrical stimulation, and ventricular tachycardia elicited by intravenous injection of ouabain or digoxin. The compound was equally effective in vagotomized or nonvagotomized dogs, and in intact cats and monkeys. The ventricular ectopic rate in conscious dogs 18 to 20 hours after two-stage ligation of a coronary artery was also markedly reduced by the drug at 2 mg/kg i.v. At antiarrhythmic dose levels, there were no undesirable effects noted on peripheral blood pressure, heart rate or the configuration of the electrocardiogram.


Phospholipids and phospholipid-bound fatty acids and aldehydes of spermatozoa and seminal plasma of rhesus monkeys.

The major components of the phospholipids of rhesus monkey spermatozoa are phosphatidyl choline (33%), phosphatidyl ethanolamine (25%), ethanolamine plasmalogen (16-1%), sphingomyelin (8-1%), choline plasmalogen (6-9%) and cardiolipin (4-5%). The major phospholipid-bound fatty acids are 16:0, 18:0, 18:1 and 22:6; the major fatty aldehydes are 15:0, 16:0 and 18:2. The same phospholipids are also present in the seminal plasma.


Effect of thyrotrophin-releasing hormone on serum prolactin levels in men with azoospermia.

Infertile men with azoospermia and low testosterone levels because of Klinefelter's or Sertoli cell-only syndrome responded to a single injection of TRH by an increase in serum prolactin levels. The degree of this response was not as great as in fertile men with normospermia and normal testosterone levels, although initial prolactin levels had been similar in both groups. The results demonstrate a link between testosterone and prolactin levels in fertile and infertile men.


Cytogenetic observations on XX/XY chimaeras and a reassessement of the evidence for germ cell chimaerism in heterosexual twin cattle and marmosets.

Testicular preparations were obtained from 7 bulls, twins of freemartins, and 1 male marmoset, all proved XX/XY chimaeras. X and Y sex chromosomes were confidently identified in nearly all the 87 spermatogonia at mitotic metaphase and 1052 primary spermatocytes at diakinesis-metaphase examined: no cell was identified as containing two X chromosomes. The germ cell chimaerism previously reported in these species is therefore not confirmed. Cultures grown from presumptive somatic these species is therefore not confirmed. Cultures grown from presumptive somatic cells in the testes of two of the bulls yielded 248 identifiable mitotic spreads, all XY-type; cultures from the gonads of their freemartin twins yielded 442 mitotic spreads, all XX-type. Direct preparations from one freemartin gonad, however, yielded 3 XY mitotic spreads out of 18 examined. The conflicting evidence concerning germ cell chimaerism in cattle and marmosets is discussed, particularly in relation to reports of XX/XY bulls that have sired a great excess of daughters. The possibility that XX germ cells contributed to the functional spermatozoa of these bulls is not favoured by present information, but is not excluded.


Molecular conformation of ammonium 8-anilino-1-naphthalenesulfonate hemihydrate. A fluorescent probe for thyroxine binding to thyroxine binding globulin.

The crystal and molecular structure of the ammonium hemihydrate salt of the fluorescent dye, 8-anilino-1-naphthalenesulfonic acid (ANS), has been determined. There are two conformationally distinct molecules in the triclinic P1 lattice. The anilino nitrogen of one molecule has slightly distorted planar geometry, and the overall conformation of the molecule is similar to that observed for the potassium salt of the fluorescent dye 2-p-toluidinyl-6-naphthalenesulfonic acid (TNS). The anilino nitrogen of the other molecule has slightly distorted tetrahedral geometry and the overall conformation of the molecule is similar to that observed for the thyroid hormones T3 and T4. The observation of two distinct conformational modifications of ANS in this crystal structure determination has shed light on the conformational flexibility of the ANS molecule itself and on the mode by which its acts as a competitive inhibitor in thyroid hormone transport proteins and as a signal for hydrophobic areas in macromolecular systems.


Structure-carcinogenic activity relationships in the Benz [a] anthracene series.

The syntheses of 1,7,12-trimethyl- and 2,7,12-trimethylbenz[a]anthracenes are described. The lack of carcinogenic activity of these compounds is discussed in relationship to the carcinogenic activity of other substituted benz[a]anthracenes.


Quantitative structure-activity relationships for 5-substituted 8-hydroxyquinolines as inhibitors of dental plaque.

Fourteen 8-hydroxyquinolines were tested for antiplaque activity by measuring their minimum inhibitory concentrations [MIC (M)] against Streptococcus mutans No. 6715. Linear regression analysis was conducted with the MIC (M) values and hydrophobic (log P), electronic (beta, pKaOH, pKaN), and steric [molar refractivity (MR), molecular weight (mol wt)] parameters. The best correlation (r2 = 0.90) was obtained with MR, log P, and beta. The smaller the steric contribution of the 5-substituent, the more active the compound. The parent 8-hydroxyquinoline was the most active. The negative contribution toward activity by 5-substituents larger than hydrogen can be overcome by the positive contributions of groups that are lipophilic and electron withdrawing; for example, the 5-chloro derivative is as active as the parent 8-hydroxyquinolines.


Azabicycloalkanes as analgetics. 3. Structure-activity relationships of 1-phenyl-6-azabicyclo[3.2.1]octanes and absolute stereochemistry of (+)-1-(3-hydroxyphenyl)-6-methyl-6-azabicyclo[3.2.1]octane and its 7-endo-methyl derivative.

A series of 53 1-phenyl-6-azabicyclo[3.2.1]octanes (1) has been tested for their analgetic and narcotic antagonist activities. Structure-activity relationships were investigated by varying the structural parameters. The most interesting compound in this series, the 1-(3-hydroxphenyl)-6,7-dimethyl derivative 8, shows the profile of a well-balanced antagonist-analgesic agent with a very mild physical dependence capacity. The absolute stereochemistry of its active enantiomer [(+)8] was established by the x-ray study and the chemical transformation to the phenylmorphan [(+)-II]. (+)-8 was stereochemically correlated also with the active enantiomer of the 7-demethyl derivatives [(+)-7] by chemical transformation and CD measurement. Certain structural and stereochemical correlations between these compounds (7 and 8) and other known antagonist-analgetics are discussed.


2-cyano-1,3-dicarbonyl compounds with antiallergic activity.

A number of 2-cyanoindan-1,3-diones and 3-cyano-4-hydroxycoumarins have been prepared and assessed for potential antiallergy activity as measured by their ability to inhibit passive cutaneous anaphylaxis in the rat, mediated by rat serum containing antigen specific IgE. The structural requirements for activity were similar not only for both series of compounds but also for the analogous 2-nitroindan-1,3-diones and 4-hydroxy-3-nitrocoumarins previously reported. The most active compounds were 2-cyano-5,6-diethylindan-1,3-dione (4e) and 3-cyano-6,7-diethyl-4-hydroxycoumarin (11h).


Inhibition of carcinogenic effects of polycyclic hydrocarbons by benzyl isothiocyanate and related compounds.

Benzyl isothiocyanate and phenethyl isothiocyanate, two compounds found in cruciferous plants, and phenyl isothiocyanate, a synthetic compound, all inhibit 7,12-dimethylbenz[a]anthracene (CMBA)-induced mammary tumor formation in female Sprague-Dawley rats when administered 4 hours prior to the DMBA. Comparable studies in which benzyl isothiocyanate was administered 24 hours before or 4 hours after DMBA showed almost complete loss of inhibition. Additions of benzyl isothiocyanate or phenethyl isothiocyanate to a diet containing CMBA inhibited formation of neoplasms of the forestomach and pulmonary adenomas in female ICR/Ha mice. Addition of benzyl isothiocyanate to a diet containing benzo[a]pyrene also inhibited carcinogenesis of the mouse forestomach due to this carcinogen. The finding of two additional anutrient dietary compounds which inhibit chemical carcinogenesis focuses on the possibility that dietary constituents of this nature may diminish the impact of exposures to chemical carcinogens.


Initiation and termination mutants of Bacillus subtilis bacteriophage SPO1.

Mutants affected in cistrons 21 and 32 of bacteriophage SPO1 are defective specifically in the initiation of DNA replication. Mutations in cistron 32 also specifically affect the termination of replication.


Selective screening procedure for the isolation of heat- and cold-sensitive, DNA replication-deficient mutants of bacteriophage SPO1 and preliminary characterization of the mutants isolated.

A procedure is described for the selective isolation of temperature-sensitive replication-deficient mutants of Bacillus subtilis phage SPO1. A modification of the procedure permits the isolation of temperature-sensitive mutants in specific cistrons of interest. The applicability of these procedures to other viral systems is discussed. The mutations isolated were assigned to eight replication-deficient cistrons, with the cold-sensitive mutations showing a distribution strikingly different from that of the heat-sensitive mutations. As a preliminary to the identification of initiation-deficient mutants, the mutants were divided into three classes on the basis of their ability to synthesize DNA after a shift to nonpermissive temperature. We also report two incidental results: (i) the SPO1 dUMP hydroxymethylase, like the T4 dCMP hydroxymethylase, may be part of a multifunctional complex; and (ii) mutants were isolated that were replication positive but lysis deficient and failed to complement one of the replication-deficient mutants.


Bacillus subtilis DNA polymerase III is required for the replication of DNA of bacteriophages SPP-1 and phi 105.

The replication of the Bacillus subtilis bacteriophages SPP-1 and phi 105 is sensitive to 6-(p-hydroxyphenylazo)-uracil (HPUra), a selective inhibitor of replicative DNA synthesis of B. subtilis which acts specifically at the levels of a replication-specific polymerase, DNA polymerase III (pol III). The origin of the HPUra-sensitive polymerase required for phage replication was examined by comparison of the drug sensitivity of phage development in a normosensitive host with that in a host carrying azp-12, a polC mutation that specifies production of an HPUra-resistant pol III. azp-12 specified HPUra-resistant phage host pol III. The host polIII requirement for SPP-1 replication also was confirmed by the demonstration that phage development was temperature sensitive in a host mutant carrying the polC mutation mut-1 (ts). Examination of the pol III activity of crude and purified cell-free preparations derived from phage-infected cells did not indicate any detectable changes in the specific activity, purification behavior, or drug sensitivity of the enzyme.


Temperate Bacillus subtilis bacteriophage phi 3T: chromosomal attachment site and comparison with temperate bacteriophages phi 105 and SPO2.

The temperate Bacillus subtilis bacteriophage phi 3T contains within its genome a locus, designated thyP3, that encodes for a protein with thymidylate synthetase activity. Bacteriophage phi 3T is different from the two previously characterized temperate phages, phi 105 and SPO2, in: heteroimmunity, response to bacteriophage antisera, endonuclease digestion pattern, induction in the presence of 6-(p-hydroxyphenylazo)-uracil, and effect on the lytic cycle of bacteriophage phi 1. The mean burst size of phi 3T is 56. The dose response curve with bacteriophage phi 3T DNA is linear for transfection and transformation to the Thy+ phenotype. The inserted prophage has been mapped by PBS1 transduction; it is between chromosomal markers ilvA8 and gltA in the terminus of the chromosome. Thus thyP3 maps at a site separate from, but between, the bacterial markers thyA and thyB when thyP3 is in the prophage state.


[TRH refractory TSH values, triiodothyronine levels and T3/T4 quotients in hypothyroid children treated with T3/T4 preparations (author's transl)].

TRH-tests, triiodothyronine (T3), TBI, and thyroxine (T4) determinations were carried out in 23 children with primary hypothyroidism treated with T3/T4-preparations. T3/T4 quotients were calculated. 7 out of 10 patients with TRH refractory TSH values and normal T4 levels had T3 values above normal range. In 6 of them pathologically elevated T3/T4 quotients could be observed. A TSH refractory TSH suppression was only found in patients with superior normal T4 values. After a reduction of the replacement doses the T3 values were normal. The findings show--that not only the T4 values should be controlled under treatment with thyroid preparations but also the T3 and TSH serum levels. The replacement therapy of hypothyroidism with T3/T4 preparations in proportion 1:4 should be revised.


Families in the treatment of alcoholism.

A review of the literature on family treatment of alcoholism suggests that although there is no single definition of family therapy, nor one single theory behind it, there is agreement that family therapy can be beneficial for both the alcoholic and the family.


Treat decision making in catastrophic illness.

Through conceptual discussion and consideration of a case study, this paper examines how physicians respond to the availability of an innovative treatment of a serious illness. It is argued that the unusual economic environment of the delivery of catastrophic illness care works with the "social contract" in medicine to encourage the use of innovative therapies, even before their efficacy has been demonstrated and often irrespective of their costs, in striking contrast with the conventional innovation adoption process. The primary constraint on catastrophic illness treatment may well be the technology or the state of knowledge. In the case of the management of leukemias in Connecticut, decisions to use drug therapies appear to have been based on a treatment trend rather than on the inherent merits of the therapies. The trend seems to have spread from the treatment of one leukemia, which responded significantly, to the management of three other leukemias. The influence on treatment decisions of a few indiviudal varibles differed across the leukemia. For example, treatment decisions in the acute leukemias were unrelated to the patient's economic status, whereas receipt of chemotherapy for one of the chronic leukemias was significantly positively correlated with economic status.


Clinical use of the anion gap.

The concepts underlying the clinical use of the anion gap (AG) and those disorders associated with its alteration are reviewed. A substantial increase in the AG usually indicates the presence of a metabolic acidosis, unless large doses of certain antibiotics or sodium salts of organic acids are being used. The etiology, pathogenesis and diagnosis of high AG metabolic acidoses are discussed. Stress is placed upon the utility of the AG in defining the cause of the acidosis, and as a guide to therapy in certain organic acidoses. A decrease in the normal AG occurs in dilutional states, hypoalbuminemia, hypercalcemia, hypermagnesemia, hypernatremia, diseases associated with hyperviscosity, bromide intoxication, and in certain paraproteinemias. The important clue provided by a low or negative AG in the diagnosis of certain of these life-threatening disorders is emphasized.


Osmotic control of the release of prolactin and thyrotropin in euthyroid subjects and patients with pituitary tumors.

The effects of acute changes in serum osmolality on basal serum PRL and TSH levels and on responses of prolactin (PRL) and thyrotropin (TSH) to the thyrotropin-releasing hormone (TRH) analogue, N3im-methyl-TRH, were studied in ten euthyroid subjects and in three patients with PRL-secreting pituitary tumors. An oral water load of 20 ml/kg had no effect on basal serum PRL or TSH levels but did result in an increased PRL response to methyl-TRH in the ten euthyroid patients. Intravenous infusion of 5% sodium chloride in the ten euthyroid subjects significantly depressed basal serum PRL levels but had no effect on the PRL response to methyl-TRH. Infusion of hypertonic saline significantly decreased the TSH response to methyl-TRH. In the three patients with pituitary tumors, oral water loading and hypertonic saline infusion had no significant effect on the basal serum PRL and TSH or the PRL and TSH responses to methyl-TRH. The patients with pituitary tumors had a higher basal serum osmolality and a proportionately higher serum concentration of arginine vasopressin than the euthyroid patients. These data suggest that changes in osmolality in euthyroid patients may have a direct effect on the anterior pituitary's PRL and TSH response to a releasing factor.


Aldehyde oxidase distribution in haltere discs of homoeotic bithorax mutants in Drosophila melanogaster.

Distribution of the enzyme aldehyde oxidase in transformed haltere discs from the homoeotic bithorax series of mutants was investigated by histochemical means. The bithorax (bx) mutant, which transforms the anterior part of the haltere into an anterior wing blade, possesses in the haltere disc an aldehyde oxidase staining pattern similar that of the anterior side of the wing disc. The postbithorax (pbx) mutant, which transforms the posterior haltere into a structure resembling the posterior wing blade, reveals an aldehyde oxidase staining pattern in the haltere disc characteristic of the posterior side of the wing disc pouch. When both (bx3 pbx) mutants are present the haltere develops into a metathoracic wing. It is shown here that the transformed haltere disc closely resembles the previously established pattern in the wing disc with respect to aldehyde oxidase distribution. Change in the pattern of aldehyde oxidase in bithorax mutants signals alteration in gene expression which at least for this particular enzyme correlates well with the morphological transformation from haltere to wing. A possible correlation between pattern of enzyme activity and developmental compartmentalization has been discussed.


[The ECG on exercise (author's transl)].

The electrocardiogram on exercise with a graduated load on the bicycle ergometer has proved of value as a screening method for coronary heart disease and arrhythmias. A horizontal or descending ST junction depression of 0.1 mv in the left precordial lead is considered a significant ischemic reaction. The present paper reports the practical performance and evaluation of findings of the method. The prognostic significance of a pathological ECG on exercise with regard to the occurrence of a coronary heart disease and the progression of the disease are pointed out.


[Behaviour of the ST segment under stress by atrial frequency and digitalis in latent coronary insufficiency (author's transl)].

Therapeutic doses of digitalis may give rise to depressions and abnormities of the ST segment. In coronary patients this is occasionally also associated with anginal complaints. In 7 patients the ST segment was investigated at rest and under increasing stress by atrial frequency without and with digitalis at increasing therapeutic dosage. In all patients a glycoside-induced, linearly intensifying depression of the ST segment was demonstrated as a regular and dose-dependent pattern the onset of which was already recognizable at an average effective level of 0,59 mg digoxin. The opinion is held that glycoside-induced ST depressions in the ECG are not in general of insignificant nature but may be the reflection of myocardial ischemia.


[Scintigraphy of the myocardium. Comparative study between thallium and perfusion scintigrams (author's transl)].

In a total of 51 patients (26 with coronary heart diseases, 25 with primary myocardial diseases), the myocardial scintigrams after intracoronary particle injection (perfusion scintigraphy) and after intravenous nuclide injection with accumulation in the myocardium (thallium scintigraphy) were compared with each other and set against the coronary angiographic and levocardiographic findings. It was shown that extensive myocardial failures (aneurysms) can be represented by both nuclear medical procedures, but that perfusion scintigraphy is more sensitive and correlates more closely to the levocardiogram findings. Smaller abnormalities of contraction of the left ventricle (hypokinetic and akinetic areas) were always shown in the perfusion scintigram, more seldom in the thallium scintigram. The outstanding advantage of thallium scintigraphy is that it is non-invasive, it can be repeated as often as desired and is easily performed during ergometric stress.


[Sources of error in the preparation and interpretation of echocardiograms (author's transl)].

Diagnostic ultrasound is able to provide important information on cardiac structure and motion. Diagnostic errors and misinterpretations however are not uncommon, due to the manner in which data are acquired and recorded during the echocardiographic examination. This paper discusses the characteristic echocardiographic patterns of a variety of important changes in the mitral valve, the left ventricle, and the interventricular and interatrial septum as well as the influence of respiration on the performance of echocardiography. Essential pitfalls in establishing a correct diagnosis in each of these conditions are emphasized and supported by illustrations.