Uniparental inheritance of mitochondrial genes in yeast: dependence on input bias of mitochondrial DNA and preliminary investigations of the mechanism. In Saccharomyces cerevisiae, previous studies on the inheritance of mitochondrial genes controlling antibiotic resistance have shown that some crosses produce a substantial number of uniparental zygotes, which transmit to their diploid progeny mitochondrial alleles from only one parent. In this paper, we show that uniparental zygotes are formed especially when one parent (majority parent) contributes substantially more mitochondrial DNA molecules to the zygote than does the other (minority) parent. Cellular contents of mitochondrial DNA (mtDNA) are increased in these experiments by treatment with cycloheximide, alpha-factor, or the uvsp5 nuclear mutation. In such a biased cross, some zygotes are uniparental for mitochondrial alleles from the majority parent, and the frequency of such zygotes increases with increasing bias. In two- and three-factor crosses the cap1, ery1, and oli1 loci behave coordinately, rather than independently; minority markers tend to be transmitted or lost as a unit, suggesting that the uniparental mechanism acts on entire mtDNA molecules rather than on individual loci. This rules out the possibility that uniparental inheritance can be explained by the conversion of minority markers to the majority alleles during recombination. Exceptions to the coordinate behavior of different loci can be explained by marker rescue via recombination. Uniparental inheritance is largely independent of the position of buds on the zygote. We conclude that it is due to the failure of minority markers to replicate in some zygotes, possibly involving the rapid enzymatic destruction of such markers. We have considered two general classes of mechanisms: (1) random selection of molecules for replication, as for example by competition for replicating sites on a membrane; and (2) differential marking of mtDNA molecules in the two parents, possibly by modification enzymes, followed by a mechanism that "counts" molecules and replicates only the majority type. These classes of models are distinguished genetically by the fact that the first predicts that the output frequency of a given allele among the progeny of a large number of zygotes will approximately equal the average input frequency of that allele, while the second class predicts that any input bias will be amplified in the output. The data suggest that bias amplification does occur. We hypothesize that maternal inheritance of mitochondrial or chloroplast genes in many organisms may depend upon a biased input of organelle DNA molecules, which usually favors the maternal parent, followed by failure of the minority (paternal) molecules to replicate in many or all zygotes.


[Genetic effects of the decay of tritium incorporated into Saccharomyces cerevisiae yeast cells. I. Lethal effect of 3H-alanine on cells of different ploidy and radiosensitivity]. Lethal effect of 3H-alanine on cells of different ploidy (1n-4n) and radiosensitivity wild type haploid and diploid strains; xrs2 mutant and diploid strains homozygous for this mutation is studied. It is shown that the efficiency of the inactivation per tritium decay depends on ploidy, radiosensitivity and geometric dimension of cells and nuclei. Do for all the studied strains is slightly higher than Do for these strains in case of external beta-irradiation. RBE of beta-ray tritium with respect to beta-ray of 32P is 1.5.


Hybrid plasmids containing the araBAD genes of Escherichia coli B/r. The DNA fragments generated by restriction endonuclease BamI which contain the araCBAD genes from E.coli B/r have been cloned. The DNA fragments containing ara genes were idenified by a compairson of the BamI fragments of lambdah80dara phages containing different ara deletion mutations. The ara genes were cloned into the plasmid pBR317, a derivative of ColE1. The cloned DNA fragments were analyzed by digestion with pairs of restriction endonucleases to determine the molecular weight of the chimeras and to identify the cloned ara DNA fragments. The cloned ara fragments were also identified by genetic complementation and recombination tests.


Cloning of chemically synthesized lactose operators. II. EcoRI-linkered operators. A 40 base, mainly duplex DNA segment, with the following sequence pAATTCCACATGTGGAATTGTGAGCGGATAACAATTTGTT (3') GGTGTACACCTTAACACTCGCCTATTGTTAAACACCTTAAp (5') has been synthesized by combination of chemical and enzymatic methods. It consists of a wild-type lactose operator sequence (boxed) bracketed by "linker" sequences which permit excision of the segment from plasmid vehicles by the EcoRI restriction endonuclease. This segment has been ligated into the pMB9 plasmid and the resulting operator plasmids used to transform E. coli K-12. Among the transformant products were strains carrying plasmids with one, two, three, or four operator segments in tandem. Derepression of the lactose operon effected by these plasmids in vivo as well as the lifetimes of complexes formed between repressor and these plasmids in vitro increase with increasing numbers of operators per plasmid.


Isolation of the regulatory segment of the biotin operons of Escherichia coli K-12. A DNA fragment of 463 base pairs has been isolated by HpaII digestion of the EcoRI cleaved lambdabiot124-10 DNA fragment containing the Escherichia coli biotin gene cluster. This HpaII subfragment contains the biotin regulator region. The biotin regulator DNA has been characterized by electron microscopic heteroduplex analysis, RNA polymerase binding capacity, and the ability to initiate leftward and rightward transcriptions. The two RNA polymerase binding sites are located 63 base pairs and 117 base pairs away from the left end of the HpaII fragment. Both the left- and rightward RNA transcripts have adenosine triphosphates at their 5' end.


A thermoinducible lambda phage-ColE1 plasmid chimera for the overproduction of gene products from cloned DNA segments. Two segments of lambda have been cloned into the multicopy plasmid pBR322. One extends from N through cII (NcII segment, from 71.3 to 81.0% on the physical map) and the other from N through P (NOP segment, from 71.3 to 86.5% on the physical map). Cells carrying these recombinant plasmids express lambda immunity (cIts) and Rex function. In addition, they decrease the efficiency of plating at 32 degrees C of lambdavir and lambdaimm434, but not that of lambdaimm21. Recombinant plasmids with lambdaNOP segments (pKC14, pKC16) differ from recombinant plasmid with labmdaNcII segment (pKC10) in two respects: (i) strains carrying pKC14 or pKC16 are killed at 42 degrees C, and (ii) these strains are thermally inducible for plasmid DNA synthesis, resulting in increase of plasmid copy number from an uninduced level of 50 to more than 130 per chromosome. It was suggested that both these differences are related to functions contained in the lambda DNA segment extending from 81.0 to 86.5%. The usefulness of plasmid pKC16 for overproduction of gene products from cloned DNA segments was demonstrated by cloning the E. coli exonuclease III gene (xth) in pKC16. Thermal induction of this xth plasmid (pSGr) results in a 125-fold increase in exonuclease III activity over that of a control strain lacking the xth gene insert. The extent of exonuclease III overproduction obtained by cloning xth gene in a lambda vector was similar to that obtained with pSGR3.


Human pituitary growth hormone cells in old age. Pituitaries obtained at autopsy from 18 men and 17 women, over 80 years of age, who died of various acute illness, were investigated histologically by various staining procedures, including the immunoperoxidase technique. Compared with pituitaries of 10 male and 10 female subjects who died of short-lasting diseases between 20 and 56 years of age, no involution of growth hormone cells was found in pituitaries of the older group. Incidence, distribution, granulation and immunoreactivity of growth hormone cells showed no apparent differences related to age and sex. Lack of regression of growth hormone cells with advancing age indicates that the pituitary can produce growth hormone in old subjects.


Home health care for the elderly.

Home health care for the elderly. Home health services must be expanded and must include ongoing social work services. In addition to the provision of direct service, social workers are encouraged to advocate for quality home health services, to develop effective techniques for providing social services to home-care patients, to incorporate these techniques into social work education, and to become involved in the education of their medical colleagues. In this way, the home health field can begin to fulfill its goals by providing patients with quality care in the home setting, thus allowing them to remain independent and at the same time meeting their health and social needs.


Diabetic patients on hemodialysis.

Diabetic patients on hemodialysis. In general, individuals who have a chronic illness must make major emotional and physical adjustments. Do patients forced to cope with two chronic conditions make these adjustments with greater difficulty or with increased ease? This study compared diabetic and nondiabetic patients having end-stage renal disease and examined how individuals already ill with diabetes adapt to the additional stress of hemodialysis treatment.


Fatal experimental Babesia microti infections in the Norwegian lemming, Lemmus lemmus (L.). Experimental infections of Babesia microti in laboratory-reared Clethrionomys glareolus revealed that approximately 15% of the erythrocytes were infected with single ring forms during peak parasitemia. Infected erythrocytes could be detected in blood smears up to one month post infection. C. glareolus treated with a single injection of Depo-Medrol i.m. two days prior to infection displayed a four-fold increase in number of infected erythrocytes at peak parasitemia, 35% of which contained more than one Babesia, and a prolongation of the infection. B. microti infections in 35 laboratory--reared Lemmus lemmus were fatal. Multiple invasion of erythrocytes, anemia, icterus, hemoglobinuria, anorexia and weight loss, and adrenal and splenic hypertrophy were characteristic for B. microti infections in Norwegian lemmings.


Application of R-plasmid DNA's from Escherichia coli minicells in genetic transformation. Components of minicell lyzates of Escherichia coli P678.54 (R1 drd 19) and escherichia coli P678.54 (R6K) were visualized in an electron microscope and used for the transformation of Escherichia coli JC7623. The frequency of the resulting transformants (of the order of 10(-6)%) was not appreciably influenced by the manner of lyzate preparation. The presence of covalently closed circular DNA was detected in two different transformants using radioisotopes, thus demonstrating an autonomous existence of R1 drd 19 or R6K plasmids in tested transformants. This finding corresponds with the results of their genetic analysis.


Stimulation of amino acid transport in Saccharomyces cerevisiae by metabolic inhibitors. Inhibitors of energy metabolism (3-chlorophenylhydrazonomalononitrile, antimycin A, iodoacetamide, dicyclohexylcarbodiimide) but not of transport (uranyl ions) stimulate at low concentrations the uptake of L-leucine, L-glutamic acid, L-arginine and, to a lesser degree, of 2-aminoisobutyric acid in Saccharomyces cerevisiae. The effect is apparent only after augmenting the energy reserves of cells by preincubation with D-glucose or, more strikingly, with ethanol. It is absent in a mutant (op1) lacking the translocation system for ADP--ATP in mitochondria. The presence of two different energy reserves for amino acid transport is indicated (one in energy-poor, the other in energy-rich cells). The stimulating effect appears to be caused by a retarded degradation of the transport proteins as occurs at a lowered level of mitochondria-produced ATP.


Cellular response in Salmonella typhimurium-infected mice: evaluation of Salmonella receptors of B lymphocytes. The cellular response in the course of experimental infection with Salmonella typhimurium was studied in mice. T cells were detected by the presence of theta-antigen, B cells by the binding of fluorescent immunoglobulins, and cells with receptors by labeled Salmonella binding. Lymphocytes were from spleen and lymph nodes. Results have been divided into three groups: group A, including mice with slight symptomatology; group B, including those with serious infection symptomatology; and group C, including mice that died in the course of the experiment. In spleen and lymph nodes of group A mice, an increase in the percentage of T and B lymphocytes was observed. This increase reached a peak 10 days after experimental infection. In lymph nodes, the B-cell percentage was equal to the percentage of T cells, whereas in spleen lymphocytes the B-cell percentage was higher. In spleens of group B mice we observed the same response as in mice of group A, whereas in lymph nodes there was a low response of T and B lymphocytes. In group C mice, there was no significant response of T and B lymphocytes in either spleen or lymph nodes. In B lymphocytes prepared from spleens of surviving mice, a small number of Salmonella receptors was detected: 200 bacterial cells per 10(9) lymphocytes.


Polymorphonuclear neutrophil chemotaxis under aerobic and anaerobic conditions. The motility of human polymorphonuclear neutrophils was studied in vitro under aerobic and anaerobic conditions. Chemotactic factors were generated from plasma with immune complexes or with whole bacteria (Staphylococcus aureus, Escherichia coli, and Bacteroides fragilis). Chemotaxis induced by chemotactic factors generated from immune complexes was identical under both conditions. However, chemotaxis utilizing chemotactic factors generated from bacteria was markedly depressed under anaerobic conditions. Mean random tubemoltility was not significantly different under aerobic and anaerobic conditions. These data indicate that different metabolic pathways may be involved in polymorphonuclear neutrophil movement. Some of these pathways require oxygen (chemotaxis in response to factors generated by bacteria in plasma), whereas others do not (random tube migration and chemotaxis in response to factors generated by immune complexes in plasma). These observations may be important in the induction of inflammatory responses within hypoxic tissues.


In vitro adhesion of Escherichia coli to porcine small intestinal epithelial cells: pili as adhesive factors. Escherichia coli strains with pili (K99 or 987P) known to facilitate intestinal colonization adhered in vitro to porcine intestinal epithelial cells. These strains adhered equally to both ileal and jejunal epithelial cells. A laboratory E. coli strain that has type 1 pili also adhered to porcine intestinal epithelial cells. When nonpiliated cells derived from 987P+, K99+, or type 1 pilus+ strains were used for in vitro adhesion assays, they failed to adhere. The attachment of piliated bacteria to epithelial cells was a saturable process that plateaued at 30 to 40 bacterial cells attached per epithelial cell. Competitive inhibition of bacterial cell attachment to epithelial cells with purified pili showed that only purified 987P competed against the 987P+ strain and only purified type 1 pili competed against the type 1 pilus+ strain. Competition between a K99+ strain and K99 was not consistently achieved. K99+, 987P+, and type 1 pilus+ bacteria could be prevented from adhering to epithelial cells by Fab fragments specific for K99, 987P, or type 1 pili, respectively. Fab fragments specific for non-K99 bacterial surface antigens did not inhibit adhesion of the K99+ strain. It is concluded that adhesion of E. coli to porcine intestinal epithelial cells in vitro is mediated by pili and that the epithelial cells used apparently had different receptors for different pili.


Characterization of an Escherichia coli plasmid encoding for synthesis of heat-labile toxin: molecular cloning of the toxin determinant. P307 is a plasmid isolated from a strain of Escherichia coli that was responsible for an outbreak of diarrheal disease in piglets. This 60 X 10(6)-dalton plasmid was subsequently shown to encode for the synthesis of a heat-labile toxin (LT). Using recombinant DNA technology, we isolated a 7.8 X 10(6)-dalton DNA fragment that contains the LT gene(s). This fragment was generated using an EcoRI partial digestion of P307 DNA, and the fragment was joined to a small multicopy plasmid, RSF2124. E. coli strains harboring the chimeric plasmid produced greater amounts of LT than did the same strains containing P307. The LT genes were also isolated on a 5.8 X 10(6)-dalton DNA fragment made by BamHI digestion, and we identified an EcoRI recognition sequence that is located in a position essential for LT synthesis.


Inhibition of specific amino acid uptake in Candida albicans by lysosomal extracts from rabbit alveolar macrophages. Lysosomal-rich fractions, obtained from normal rabbit alveolar macrophages, were extracted and tested for their effects on Candida albicans. The uptake and incorporation of various compounds (amino acids, uridine, 2-deoxy-D-glucose, and Rb+) by C. albicans were measured in the presence and absence of extract. These studies demonstrated that the extract had a specific effect on the uptake of certain amino acids by C. albicans. Of the amino acids tested, the uptake of methionine valine, lysine, phenylalanine, and leucine was drastically reduced in the presence of extract, whereas proline and glutamic acid uptake was unaffected. Those amino acids whose uptake was inhibited have been shown to be transported in other yeasts by a general amino acid permease. The existence of a general amino acid permease in C. albicans is compatible with our data. Additionally, the extract had no effect on the uptake of uridine, 2-deoxy-D-glucose, and Rb+. Leakage of 86Rb by C. albicans was detected in the presence of the extract. Viability of Candida, as measured by colony-forming ability, decreased after a 16-h incubation of C. albicans with the extract.


Biological evaluation of a methanol-soluble, heat-stable Escherichia coli enterotoxin in infant mice, pigs, rabbits, and calves. Escherichia coli P16 was shown to produce two heat-stable toxins (ST) with differing biological activity. The toxins were separated by methanol extraction, and the first, STa, was methanol soluble, partially heat stable, active in neonatal piglets (1 to 3 days old) and infant mice, but inactive in weaned pigs (7 to 9 weeks old); the second, STb, was methanol insoluble, active in weaned pigs and rabbit ligated loops, but inactive in infant mice. It is therefore suggested that use of suckling mice as indicators of ST production will fail to identify certain ST-producing strains.


Increase in the population of duodenal immunoglobulin A plasmocytes in axenic mice associated with different living or dead bacterial strains of intestinal origin. Various bacterial strains were tested for their ability to stimulate immunoglobulin A (IgA) plasmocytes to populate the duodenal lamina propria in axenic mice. The mice were associated with the strains for at least 4 weeks. The strains inhabiting the conventional mouse intestine and belonging to the genera Lactobacillus, Streptococcus, Eubacterium, Actinobacillus, Micrococcus, Corynebacterium, and Clostridium (including the extremely oxygen-sensitive ones) are only slightly or nonimmunogenic, whereas the strains belonging to the genera Bacteroides and Escherichia have an immunogenic effect. The same result was obtained with Bacteroides and Escherichia strains isolated from the digestive tract of other animal species. The kinetics of appearance of intestinal IgA plasmocytes are similar in axenic mice monoassociated with a stimulatory strain and in conventional mice. The association of two or more strains with axenic mice leads either to the same or a greater number of duodenal IgA plasmocytes as that obtained with the most stimulatory strain monoassociated with axenic mice. The maximum stimulation recorded in all of these trials represents about two-thirds of that observed in conventional mice and was obtained in the duodenum of gnotoxenic mice harboring four bacterial strains isolated from the conventional mouse microflora. The orally administered killed cells of two immunogenic strains, E. coli and Bacteroides sp., are as immunogenic as the living cells, provided that their concentration in the digestive tract is sufficient.


Extraction of streptococcal type 12 M protein by cyanogen bromide. Conditions for the release of streptococcal type 12 M protein from whole cells by cyanogen bromide are described; they demonstrated that methionine is not essential to the structural arrangements which account for some of its immunological and biological properties. The released M protein was separated from other proteins by column chromatography with hydroxylapatite. The type-specific molecules which reacted with precipitating antibodies were found only in the 0.3 M eluate, formed zones with mobilities less than 12% of that of the dye front on electrophoresis in the standard acrylamide disc gel system, formed at least four bands in sodium dodecyl sulfate-acrylamide disc gels with molecular weights ranging from 12,000 to 23,000, and stimulated the formation of opsonic antibodies in rabbits. Cyanogen bromide provides a highly specific method for the release of M proteins which should prove particularly useful in analyses of structural-functional relationships among different M proteins.


Comparative immunological analysis of host plasma proteins bound to bloodstream forms of Trypanosoma brucei subspecies. The presence, location, host specificity, identity, and quantity of rat plasma proteins bound to bloodstream forms of Trypanosoma brucei subsp. brucei, T. brucei subsp. rhodesiense, and T. brucei subsp. gambiense were determined by a quantitative indirect fluorescent-antibody method and gel immunoassays. Fluorescence differences between trypanosomes obtained from rats and mice and treated with antiserum to normal rat plasma indicated that most, if not all, of the bound plasma proteins were host specific. Removal of plasma proteins by trypsinization of parasites provided evidence for their attachment to the surface of the parasite. The accreted proteins were found to be host albumin, immunoglobulin G (IgG), and complement (C3). The same quantities of these three plasma proteins were present on T. brucei subspecies collected from normal rats at 2 days postinfection, during low or peak parasitemias, or from cortisone-treated rats. IgM could only be detected on parasites collected from normal rats at peak parasitemia. With the aid of rocket immunoelectrophoresis, host albumin and IgG were found to account for 0.2 and 0.05%, respectively, of the total soluble proteins of the bloodstream forms. It was concluded from this study that: (i) host plasma proteins were bound to parasites early in the infection, suggesting a mechanism of adaptation to the mammalian host; (ii) the surface-bound IgG was not the result of a specific immune response against the parasites but might be the cause of C3 attachment; (iii) among the bloodstream forms of the three T. brucei subspecies, there were no differences in amounts of surface-bound albumin, IgG, or C3. A comparison between the present data dealing with T. brucei subspecies, on the one hand, and the previously published results concerning T. congolense, on the other, revealed significant differences in the amounts of the host plasma proteins attached to these hemoflagellates.


Effects of certain cations (Fe, Zn, Mg, and Ca) on bacterial endotoxins. The natural occurrence of cations Fe, Zn, Mg, and Ca in the lipopolysaccharide (LPS) of both the S and R forms of Shigella dysenteriae 1 was studied. LPS preparations were obtained either by phenol-water extraction (according to the method of Westphal et al., Z. Naturforsch. 7b:148-155, 1952) or by extraction of cells with hypertonic sodium chloride-sodium citrate (according to the method of Raynaud and Digeon, C. R. Acad. Sci. (Paris) 229:564-566, 1949), with subsequent chromatographic purification on Sephadex G200 and Sepharose 4B columns. The cation in highest concentration in the Westphal extract was Mg(2+) (as much as 30 mug/mg), and the lowest one was Fe (ca. 0.10 mug/mg). In LPS of the Raynaud type, the cation in highest concentration was Ca(2+) (as much as 13 mug/mg), and the lowest one was Fe (ca. 0.10 mug/mg). The effects of increasing and decreasing the concentrations of cations (Fe, Zn, Mg, Ca) upon the biological activity of the endotoxins was evaluated by using toxicity in mice and the Limulus test. It appeared that increased concentrations of Fe (chiefly of Fe(3+)) decreased the toxicity of the R form of LPS, whereas Mg(2+) decreased the toxicity of the S form. After prolonged dialysis of LPS preparations against deionized water, there was no consistent relationship between toxicity as determined in white mice and with the Limulus test.


Homogeneous enzyme immune assay for staphylococcal enterotoxin B. A simple homogeneous enzyme immune assay was developed for detection of staphylococcal enterotoxin B by a conjugate of beta-amylase coupled with the enterotoxin.


Cefazolin and cephradine: relationship between serum concentrations and tissue contents in mice. The relationship between serum concentrations and tissue levels of various doses of cefazolin and cephradine was determined in experimentally infected mice. An infection was induced by injection of 5 X 10(6) Escherichia coli into the right hind leg, antibiotics were administered 1 h later. At 15-minute intervals mice were killed by exsanguination after a blood sample was taken, and the infected thigh was taken out and homogenized. The total amount of antibiotic per gram thigh muscle tissue was calculated on the basis of the concentration in the supernatant of the homogenate. From the parallel course between blood concentrations and total tissue contents it may be concluded that the thigh was easily accessible for both cephalosporins. The ratio between the total tissue content and the free serum concentrations was the same for both antibiotics despite the difference in protein binding between cefazolin and cephradine in mouse serum. This implies that the discrepancy between the relative antibacterial activity of the cephalosporins in vitro and in vivo found in earlier experiments cannot be explained by differences in accessibility of the tissue.


Continuous monitoring of urea levels during hemodialysis. An ammonium ion-specific electrode system is evaluated for analysis of blood urea nitrogen from serum, plasma ultrafiltrate, or hemodialyzer dialysate fluid. The electrode shows a high sensitivity over clinically useful concentration ranges. Free serum ammonia, volatile amines, or urea ammonia after hydrolysis can be measured. In a double blind study excellent correlation was found for 187 blood urea nitrogen samples measured with the electrode with standard auto-analyzer techniques. Continuous monitoring of urea in dialysate fluid is described. Urea clearances were measured from dialysate effluent from an in vitro dialysis using conventional equipment. These values show excellent correlation to those from stnadard analysis methods. A continuous urea sensor for on-line hemodialysis application which can provide quantification of therapy is described.


The principles of controlled clinical trials of drugs. The principles of evaluation of new drugs are described and discussed. This evaluation is carried out in three consecutive phases. The first one is performed on young, healthy volunteers. The general response to the drug and signs of toxicity are registered. Preliminary data about absorption, elimination and metabolism are collected. The safe dosage range and preferred route of administration are settled. In the second phase, initial trials on a limited number of patients for specific disease control or prophylaxis are carried out. Patients are included in the trial by random selection and stratification. Studies are strictly controlled. To avoid bias, double-blind examination is used in paired patients, a group of patients or by the crossover design. The problems of planning, design of the study, clinical and statistical evaluation of results are discussed. Two examples of controlled clinical studies are given, concerning the assessment of antirheumatic and antianginal drugs. Finally the principles of the third phase of study are discussed. The methods of evaluation described may be applied for the assessment of entirely new substances as well as for verification of the objective therapeutic value of "old" drugs.


Methodological aspects of clinical evaluation of antianginal drugs. The principles of patient selection and designing and management of the clinical evaluation of antianginal agents are discussed. The study should involve patients with stable angina pectoris, history of anginal attacks of at least three months duration with five or more attacks per week, ST-segment depression at least one mm and anginal attack during the exercise test. Subjective criteria based on detailed dilaries kept by the patients and objective criteria based on exercise tests with cycloergometer, treadmill and atrial pacing are described; and difficulties related to final evaluation of the antianginal agent are discussed.


Pharmacokinetics and therapeutic efficacy of metronidazole at different dosages. Metronidazole, a drug effective against certain protozoal and anaerobic infections, was given female patients with Trichomoniasis urogenitalis. Group I received twice daily 250 mg of metronidazole (supplied as 250 mg tablets Vagimid). Group II received in a single dose 1.0 g (4 tablets); and group III, 2.0 g (8 tablets). Serum and urine metronidazole levels were measured polarographically. Kinetic parameters were determined from the measured values of the concentration time curve by a computing program. An exact control of the therapeutic result was carried out. In all patients peak serum levels occurred within 1-3 hr and averaged 5.1 +/- 1.7 microgram/ml after 250 mg doses, 19.6 +/- 3.8 microgram/ml after 1.0 g doses and 40.6 +/- 9.3 microgram/ml after 2.0 g doses. About 35% of the administered dose was recovered in the urine in 12 hr and about 50% in 24 hr. Metronidazole shows protein binding of 10-20% equally in vivo and in vitro. Minimum trichomonacidic concentrations of nearly 1 microgram/ml were still present 12 hr after oral application of 250 mg metronidazole, and 24 hr to 36 hr, respectively after 1.0 g and 2.0 g daily doses. The cure rate was 100%. No serious side effects ocurred in any of the patients.


The synthesis of DNA by DNA-membrane complexes from irradiated E. coli B/r and Bs-1: the role of the membrane. DNA-membrane complexes were isolated from lysed E. coli B/r and Bs-1, either by low g forces from a low salt solution, or by high g forces through a discontinuous sucrose gradient. The latter method was more gentle. Irradiation of the intact bacteria had no effect on the membrane macromolecules or on RNA components of these complexes. DNA loss was not significant after irradiation under anoxic conditions but complexes isolated from from Bs-1 irradiated in air showed an appreciable decrease in DNA content. In the presence of the appropriate nucleotide mixture, both 'free' DNA, found in the supernatant fractions, and rapidly sedimented membrane-associated DNA were able to synthesize DNA in the absence of added polymerase. DNA synthesis associated with 'free' DNA was more sensitive to radiation than that associated with DNA bound to the membrane, which appeared to moderate the effects of radiation on new DNA synthesis. It is concluded that the depression of DNA synthesis is primarily a result of irradiation-induced changes on genome-DNA. The interpretation of earlier work from our laboratories that DNA-membrane complexes contained the macromolecular structure which responded to radiation with a high o.e.r. is not supported by the evidence in this work.


Effect of hypervitaminosis A on the Harderian gland in rats. A morphologic and morphometric study. Daily administration of 100 I.U. vitamin A/gram body weight to rats, for a period of 15 days, had the following effects: Body weight of rats treated with excess vitamin A was lower (84.0 g) than that of the controls (147.0 g). The weight of the Harderian gland was lower of rats injected with vitamin A (54.0 mg) than for the controls (87.0 mg). Histological examination of the Harderian gland in the hypervitaminotic animal showed smaller acini, lower cells, wide lumina and small nuclei when compared with the controls. These data were confirmed with morphometric techniques.


Plasma levels of folates, ascorbate, vitamin B6 and riboflavin in children. Plasma vitamin levels of approximately 125 children admitted to the Queens Child Psychiatric Center were determined in a controlled study, with respect to medication and dietary conditions. The age of these children varied from 4 to 16 years. The data present the mean values with respect to the age and racial origin. There are no significant variations in the mean levels of these vitamins in terms of the age etc. of the subjects. As such the data indicate the levels in children and are comparable to the data for adults.


Ascorbic acid, neutrophil function, and the immune response. The role of ascorbic acid is reviewed with regard to antimicrobial activity, interferon production, and humoral and cellular immune responses. Ascorbic acid appears to play a role in a number of neutrophil functions including increased chemotaxis, increased particulate ingestion, enhanced lysozyme-mediated non-oxidative killing, protection against the toxic effects of superoxide anion radical, inhibition of the halide-peroxide-myeloperoxidase system without a pronounced bactericidal effect, and stimulation of the hexose monophosphate shunt.


Bacterial catabolism of lipoic acid. Isolation and identification of a methyl ketone. A soil bacterium, Pseudomonas putida LP, can be grown on lipoate as sole source of carbon, sulfur, and energy. In addition to the previously identified catabolites (bisnorlipoate, tetranorlipoate, beta-hydroxybisnorlipoate, lipoate thiolsulfinate, and two bisnorlipoate thiolsulfinates) isolated from cultures of the organism grown on [1,6-14C[lipoate, a methyl ketone (1,2-dithiolane-3-butyl-3'-one) has now been isolated and identified. This catabolite was isolated by solvent extraction and hydrophobic gel filtration and characterized by chromatographic mobilities and spray reactions and by UV, IR, PMR, and mass spectrometries. The methyl ketone presumably arises by decarboxylation of the beta-keto acid formed during the beta-oxidation of lipoate to bisnorlipoate by the microorganism.


Glomerular foam cells in kidney allograft. Foam cells are occasionally encountered in renal glomeruli in cases of nephrotic syndrome, however, they were not described so far in renal allografts. The authors found the glomerular foam cells in a cadaveric kidney transplant which survived 7 months after transplantation in a 35-year-old man. Electron microscopy revealed the mesangial cells to be loaded with lipids in this case. The formation of the glomerular foam cells is generally referred to hyperlipidemia, but its cause remains obscure in the presented case.


A diagnostic key employing biological reactions for differentiating pathogenic Corynebacterium genitalium (NSU corynebacteria) from commensals of the urogenital tract. More than 100 strains of Corynebacterium genitalium, probably responsible for coryneform urethritis and other infections, and 600 commensals of the male and female urogenital tracts have been studied and grouped into five pathogenic types numbered I to V and six saprophytic types designated C-1 to C-6 on the basis of eight biological reactions. This preliminary classification has been based on differences in requirements for oxygen, on the fermentation of fructose, dextrose, sucrose, and starch together with the production of the enzymes gelatinase, lipase, and urease. One criterion differentiated the pathogens from the commensals: All pathogens were nonfructose fermenters whereas every commensal fermented this sugar.


99m technetium sulphur colloid localization in renal infection. An experimental evaluation. Renal uptake of 99mTechnetium sulphur colloid (99mTcSC) was investigated in 99 animals with experimentally induced unilateral pyelonephritis and in 60 animals with a thermally induced renal injury. The degree of localization of 99mTcSC in the kidney was determined by direct counting of retained isotope after bilateral nephrectomy at autopsy. Phagocytosis of 99mTcSC by inflammatory tissue has been reported and it was anticipated that renal infection would lead to an increase in 99mTcSC uptake by the kidney. In fact it was found that renal infection with pathologic changes resulted in a significant decrease in the localization of 99mTcSC in the pyelonephritic kidney. A similar decrease was found in the thermally injured kidney but renal infection without pathologic changes did not give rise to any changes in radionuclide uptake. These findings suggested that phagocytosis by the proximal convoluted tubular cells is the normal mechanism for 99mTcSC accumulation in the kidney and that renal infection reduced the phagocytic capability of these cells.


Towards computer analysis of pulmonary infiltration. A feasibility study is described to provide quantitative texture measures to distinguish between normal lung, alveolar infiltrates and interstitial infiltrates. Advanced computer imaging technology and decision making processes were applied to distinguish between these textural patterns. The results, based on computer extracted quantitative measures, show an excellent separation of the three classes considered with 95% accuracy in the training phase and 90% accuracy in the testing phase.


Lymphatic visualization during renal transplant rejection. Intrarenal microlymphatic filling was observed during a microangiographic and histologic study of unmodified canine allograft rejection. The extent of lymphatic visualization was correlated with the pathophysiologic state of the allograft. An example of renal lymphatic visualization following arterial perfusion of a human specimen is also presented. During early rejection, as microvascular and histologic alteration became more severe, there was prominent visualization of lymphatics. However, in late rejection, as cortical necrosis developed, lymphatic filling was less evident. Lymphatic visualization following arterial perfusion during rejection depends on progressive loss of microvascular integrity, contrast extravasation and subsequent filling of lymphatics which are acting as an alternate route of drainage for the excess interstitial edema present due to immune injury.


Locus of control and health: a review of the literature. Locus of control, an individual difference construct from social learning theory, has shown some promise in predicting and explaining specific health-related behaviors. Research is reviewed on the utility of the locus of control construct in understanding smoking reduction, birth control utilization, weight loss, information-seeking, adherence to medication regimens, and other health or sick-role behaviors. Implications for health educators are presented.


Evaluation of histochemical observations of activity of acid hydrolases obtained with semipermeable membrane techniques: a combined histochemical and biochemical investigation 2. The biochemical investigation and comparison with the histochemical observations. The reliability of enzyme histochemical semipermeable membrane techniques for the demonstration of acid hydrolases was investigated with a combined histochemical and biochemical study. In part 1 the histochemical findings were presented. In this communication the biochemical findings are reported and compared with the histochemical findings. In m. soleus, m. plantaris, m. gastrocnemius and diaphragm of vitamin E deficient rabbits the activity of the lysosomal acid hydrolases, cathepsin D, acid maltase, acid phosphatase and beta-glucuronidase is significantly increased. This increase in activity of the investigated acid hydrolases was equal for muscles with an aerobic or an anaerobic metabolism. By means of statistical calculations the activity of the enzymes demonstrated with histochemical techniques was compared with the enzyme activity determined with biochemical techniques. From the results of this investigation it can be concluded that the histochemical semipermeable membrane techniques for the demonstration of activity of acid hydrolases are very reliable. Considering the fact that these techniques are also tissue-saving, they are therefore extremely suitable for the study of catabolic wasting processes in skeletal muscle tissues of patients with inherited or acquired muscular diseases.


Immunofluorescent subcellular localization of some muscle proteins: a comparison between tissue sections and isolated myofibrils. The localization of parvalbumin in fish white muscle and of the calcium binding protein, of arginine kinase and of glycogen phosphorylase in crayfish tail muscle have been investigated by immunofluorescence using isolated myofibrils and muscle sections as starting materials. It is shown that the four proteins appear to be localized on the thin filaments when myofibrils are used as starting material. This result contrasts with previous observations where it appeared that parvalbumin in fish muscle and arginine kinase in crayfish muscle were distributed uniformly within the cell. This discrepancy is discussed in relation to the high solubility of these proteins. In the light of the present knowledge about striated muscles from these two organisms, it seems that the roles of parvalbumin in fish and of the calcium binding protein in crayfish are probably different.


Evaluation of histochemical observations of activity of acid hydrolases obtained with semipermeable membrane techniques: a combined histochemical and biochemical investigation 1. The histochemical investigation. The reliability of enzyme histochemical observations of activities of acid hydrolases was investigated with a combined histochemical and biochemical study. Specimens of m. soleus, m. plantaris, m. gastrocnemius and diaphragm of normal and of vitamin E deficient rabbits were used. For the histochemical investigation, activity and localization of acid phosphatase, beta-glucuronidase, leucine aminopeptidase and E600 resistant non-specific arylesterase were examined with semipermeable membrane techniques. For the biochemical investigation, activity of acid phosphatase, beta-glucuronidase, cathepsin D, acid maltase and neutral maltase was determined. By means of stastical calculations the enzyme activities demonstrated with histochemical techniques were compared with the enzyme activities determined with biochemical techniques. In the present communication the histochemical findings are reported and discussed. From the histochemical findings it appeared that activity of the acid hydrolases investigated is strongly increased in both a granular and a diffuse pattern in skeletal muscle of vitamin E deficient rabbits. The statistical calculations of the histochemical findings clearly reveal that the increased activity of one acid hydrolase was highly significantly paralleled by an increased activity of a second acid hydrolase. Moreover the probability that the activity of all other histochemically studied acid hydrolases was significantly increased was rather high. The increase in activity of the acid hydrolases studied was the same in muscles with an aerobic or an anaerobic metabolism. Moreover there was no difference in activity and localization of the acid hydrolases in aerobic type I and anaerobic type II fibres. The localization of acid phosphatase and beta-glucuronidase activity muscle fibres mostly coincided. In cases where these enzymes were localized both centrally and in the subsarcolemnal areas of the muscle fibres, the activity of E600 resistant naphtholesterase was usually, and the activity of leucine aminopeptidase was exclusively located in the subsarcolemnal areas. All of the examined acid hydrolases were found to be present in the inflammatory exudate and in the connective tissue.


A methodological study for the quantification and the control of the physico-chemical processes occurring during cell fixation. I. The development of a new technology. Fixation in a traditional sense means the immersion of biological material into a chemical fluid. For permanent preservation the fixative is always "offered" (1) in excess of the cell sample, and the process of fixation is influenced by (2) chemical impurities of the fixative fluid. Both factors influence the succeeding dyeing of cells. In order to avoid these uncontrolled criteria, a new technology for controlled cell fixation has been developed, whereby freshly prepared formaldehyde and methanol gas in an "inert" gas-flow of helium was applied to thin membranes by aid of a capillary flow-in technique. The instrumental equipment consists of (1) an ultra-high vacuum flow-apparatus with a total-pressure measuring unit, (2) a gas-supply device, (3) a mass spectrometer including a pump system, and (4) a Teflon and/or glass-gas chamber for the treatment of synthetic (Hostaphan foils) or biological membranes (mesenterium) with formaldehyde as the fixative gas. The amount of "offered", adsorbed, absorbed, diffused, and desorbed fixative gas could be absolutely estimated after the saturation of the membranes with an "on-line" operating "inert" mass spectrometer of the Omegatron type. The gas treatment of the Hostaphan foils with formaldehyde showed that nearly all adsorbed gas molecules could be desorbed. In contrast to native membranes the greatest proportion of the gas molecules adhered to the biological surface, and only a small quantity were desorbable. Physisorption or physisorption and chemisorption occured depending on the adsorber surface property. A monolayer of formaldehyde of 5.10(14) to 1.10(15) molecules per 10(16) A2 surface area can be postulated on the basis of these preliminary results. This value corresponds to a mass of about 5.10(-8) g CH2O. It resulted in an area-coverage ratio of CH2O molecules per cell of 10(9):1. The membrane surface facing the gas side always amounted to 1 cm2. A fixative gas concentration of 10(6) molecules/cm3, and therefore a degree of coverage of less than 1/1000 monolayer can be estimated absolutely. For a precise determination of the degree of fixation, further experiments and the evaluation of additional physico-chemical parameters are necessary.


Immunoenzymehistochemical detection of fibrin microthrombi during disseminated intravascular coagulation rats. In tissue of rats with disseminated intravascular coagulation, fibrin microthrombi can be sensitively detected by immunohistochemical methods, using antisera against rat fibrinogen or fibrin monomer. An indirect immunoperoxidase procedure on paraplast-embedded sections yields best results with regard to the morphology of the thrombi and their localization in the tissue. Only fibrillar immunoreactive material, oriented lengthwise in the vessels, should be regarded as microthrombi formed in vivo.


Increased production of the antibiotic aurodox (X-5108) by aurodox-resistant mutants. Conventional strain and media improvement techniques were of limited success in increasing yields of the antibiotic aurodox (X-5108) above 0.5 g/liter. Higher yields were obtained by reversion of a zero producer followed by the selection of mutants resistant to aurodox. Resistant strains of Streptomyces goldiniensis ATCC 21386 able to grow on 2 g/liter of aurodox produced greater than 2.5 g/liter of antibiotic. The rate of yield increase leveled off as the strains became resistant to greater than 2 g/liter of aurodox. These strains, in contrast to those sensitive to aurodox, gave a positive response to conventional mutagenic methods for further yield increases.


In vitro activity against Escherichia coli of CGP 9000, a new oral cephalosporin. The activity against Escherichia coli of a new oral cephalosporin (manufacturer's code: CGP 9000) has been evaluated in vitro. The intrinsic lytic activity of the new compound was greater than that of cephalexin, but less than that of cephalothin. As judged by regrowth studies using ampicillin resistant E. coli strains, the beta-lactamase stability of the new cephalosporin was somewhat less than that of cephalexin. When tested in an in vitro model in conditions simulating those of the treatment of bacterial cystitis, cephalosporin CGP 9000 suppressed growth of an ampicillin sensitive E. coli strain for a therapeutically acceptable period of time, but exhibited reduced activity against an ampicillin resistant E. coli strain.


Substrate specificity of penicillin acylase of E. coli. The hydrolysis of several phenylacetylamino compounds was studied using a purified preparation of E. coli penicillin acylase. The L-isomers of phenylacetyl amino acids were cleaved much faster than the D-isomers. The same observations was made for some phenylacetylamino beta-lactams. When the beta-lactam ring is incorporated in a penam or cephem ring system, the D-isomers were hydrolysed somewhat faster than the L-isomers. We also confirmed that benzylpenicillins with an hydroxy- or an amino-group in alpha-position of the side chain were hydrolysed, both in the normal and the 6-epi-series.


Intraperiplasmic growth of Bdellovibrio bacteriovorus 109J: N-deacetylation of Escherichia coli peptidoglycan amino sugars. During intraperiplasmic growth of Bdellovibrio bacteriovorus on Escherichia coli, the substrate cell peptidoglycan is extensively modified as it is converted to bdelloplast peptidoglycan. The initially lysozyme-sensitive peptidoglycan of E. coli was rapidly converted to a lysozyme-resistant form. The conversion was due to the N-deacetylation of a large portion of the peptidoglycan amino sugars. Chemically acetylating the isolated peptidoglycan restored its sensitivity to lysozyme digestion. However, approximately half of the products of lysozyme digestion exhibited hydrophobic interactions that were shown not to be due to the presence of protein. This suggests that a molecule capable of hydrophobic interactions, other than protein, becomes linked to the bdelloplast peptidoglycan. The data also suggest that much of the Braun lipoprotein is removed from the E. coli peptidoglycan early during bdellovibrio development.


Intraperiplasmic growth of Bdellovibrio bacteriovorus 109J: attachment of long-chain fatty acids to escherichia coli peptidoglycan. During the initial stages of intraperiplasmic growth of Bdellovibrio bacteriovorus on Escherichia coli, the peptidoglycan of the E. coli becomes acylated with long-chain fatty acids, primarily palmitic acid (60%) and oleic acid (20%). The attachment of the fatty acids to the peptidoglycan involves a carboxylic-ester bond, i.e., they were removed by treatment with alkaline hydroxylamine. Their linkage to the peptidoglycan does not involve a protein molecule. When the bdelloplast peptidoglycan was digested with lysozyme, the fatty acid-containing split products behaved as lipopeptidoglycan, i.e., they were extracted into the organic phase of 1-butanol:acetic acid:water (4:15) two-phase system; all of the lysozyme split products generated from normal E. coli peptidoglycan were extracted into the water phase. It is suggested that the function of the acylation reaction is to help stabilize the bdelloplast outer membrane against osmotic forces. In addition, a model is presented to explain how a bdellovibrio penetrates, stabilizes, and lyses a substrate cell.


Reconstitution of an ordered structure from major outer membrane constituents and the lipoprotein-bearing peptidoglycan sacculus of Escherichia coli. An ordered hexagonal lattice structure with a lattice constant of about 7 nm was reconstituted on the entire surface of the lipoprotein-bearing peptidoglycan from outer membrane protein O-8 and lipopolysaccharide. The lattice structure resembled that observed in the cell envelope which had been treated with sodium dodecyl sulfate (Steven et al., J. Cell Biol. 72:292-301, 1977). The omission of either O-8 or lipopolysaccharide resulted in the failure of formation of the lattice structure. No ordered lattice was formed on the peptidoglycan lacking the bound form of the lipoprotein. In the absence of the lipoprotein-bearing peptidoglycan, O-8 and lipopolysaccharide assembled into vesicles with an ordered hexagonal lattice, the lattice constant of which was also about 7 nm. A preliminary experiment indicated that protein O-9 gave the same result as did O-8. These results strongly indicate that O-8 and/or O-9 and lipopolysaccharide provide the ordered framework of the outer membrane and that the bound form of the lipoprotein plays a role in the holding of the framework on the peptidoglycan layer.


Cell-cell interactions in conjugating Escherichia coli: role of F pili and fate of mating aggregates. Bacterial conjugation between Escherichia coli cells was investigated by a combination of physical and genetic techniques, using Hfr, F', or R+ donors and F- recipients. DNA transfer occurred in mating aggregates of up to 50 cells. Multiple interactions between donor and recipient cells occurred, and both F- pilus connections and wall-to-wall contacts were detectable. The detectable F- pilus contacts could be destroyed without either disrupting the mating aggregates or preventing DNA transfer. Hfr X F- mating aggregates did not disaggregate even though recombinant frequencies were inversely proportional to the distance from the origin of DNA transfer. F' or R+ donors formed mating aggregates with F- cells which disaggregated soon after transfer of the autonomous sex factor DNA.


Transconjugant analysis: limitations on the use of sequence-specific endonucleases for plasmid identification. We used the sequence-specific endonucleases EcoRI, SmaI, BamHI, HsuI, and HaeIII as identification tools in following the conjugal transfer of the well-studied R plasmids Sa, R388, RP4, and R6K. Transfers were both intergeneric and intrageneric. Plasmid fingerprints were generated from both single- and combination-enzyme digests. The Sa transconjugants yielded plasmids showing consistent fingerprints for each of the respective endonucleases used, whereas the three other R-plasmid transconjugants showed fingerprint changes.


Porin activity in the osmotic shock fluid of Escherichia coli. Osmotic shock fluid of Escherichia coli exhibited pore-forming activity. This activity could be followed by an in vitro assay based on the conductivity increase for ions due to the presence of pores in black lipid membranes. The histogram (the distribution of conductivity increments in a single pore experiment) obtained with osmotic shock fluid from E. coli was identical to the histogram obtained by detergent-solubilized porin isolated from the outer membrane. The osmotic shock fluid from porin-negative mutants also exhibited pore activity, although the histogram and ion specificity were different from those of porin. Antibodies raised against detergent-solubilized porin were able to form precipitin lines by the Ouchterlony immunodiffusion technique when shock fluids, but not detergent-solubilized porin, were used. These antibodies prevented the formation of pores when shock fluids contained porin but not when shock fluids obtained from porin-negative mutants were used. Macroscopic membrane conductivity of shock fluids due to porin exhibited a concentration dependence, in contrast to detergent-solubilized porin. These results indicate that the hydrodynamic properties of periplasmic or "soluble" porin are different from those of the detergent-solubilized porin of the outer membrane. Periplasmic porin comprises about 0.7% of total protein in the osmotic shock fluid.


Identification of three genes controlling production of new outer membrane pore proteins in Escherichia coli K-12. Escherichia coli K-12 strains carrying mutations in the ompB gene or double mutations in the tolF and par genes lack the major outer membrane proteins 1a and 1b. These strains are deficient in the transport of small hydrophylic compounds and are multiply colicin resistant. When revertants of these strains were sought, a number of extragenic pseudorevertants were obtained which produced new outer membrane proteins. These new proteins could be divided into three classes by differences in electrophoretic mobility on polyacrylamide gels, by differing specificities for transport of small molecules, and by the identification of three different genetic loci for genes controlling their production. These genetic loci are designated as nmpA (at approximately 82.5 min on the E. coli K-12 genetic map), nmpB (8.6 min), and nmpC (12 min). The new proteins produced in strains carrying nmpA, nmpB, or nmpC mutations did not cross-react with antiserum against a mixture of proteins 1a and 1b, or with antiserum against phage-directed protein 2. Production of the new membrane proteins restored sensitivity to some of the colicins.


Differences in crystal violet uptake and cation-induced death among yeast sterol mutants. Yeast sterol mutants exposed to crystal violet demonstrated greater dye uptake than the wild type. In addition, exposure for 10 min to hypertonic cation solutions showed a greater decrease in cell viability for mutants than for wild type.


Mapping of the hemE locus in Salmonella typhimurium. A new type of heme-deficient mutant was isolated in Salmonella typhimurium by neomycin selection. The mutant was deficient in uroporphyrinogen decarboxylase activity, coded by the hemE gene. The hemE gene was located between the genes rif and thi at 128 min on the chromosomal map of S. typhimurium.


A single mutation affects L-serine deaminase, L-leucyl-, L-phenylalanyl-tRNA protein transferase, and proline oxidase activity in Escherichia coli K-12. A mutation at a single locus, wyb, results in several phenotypic changes in Escherichia coli K-12. The Wyb- phenotype includes: (i) an increase in L-serine deaminase activity, together with a loss of inducibility by L-leucine; (ii) an absence of L-leucyl-, L-phenylalanyl-tRNA protein transferase activity; (iii) inducibility of proline oxidase by proline; and (iv) a loss of ability to use maltose as a carbon and energy source.


Control of Escherichia coli growth by CO2. Escherichia coli B dependence on CO2 for growth was demonstrated. At suboptimal CO2 concentrations the rate of growth was controlled by CO2 concentration.


Temperature-dependent variation in the extent of methylation of ribosomal proteins L7 and L12 in Escherichia coli. The amount of epsilon-N-monomethyllysine in ribosomal proteins L7 and L12 in Escherichia coli is dependent upon the cell growth temperature. At 37 degrees C or above, very small amounts were detected. Dramatic increase in the content of epsilon-N-monomethyllysine in these proteins was observed when the growth temperature was lowered.


Distribution of autolysins in hyphae of Aspergillus nidulans: evidence for a lipid-mediated attachment to hyphal walls. Preparations of broken Aspergillus nidulans hyphae contained both free and wall-bound autolysins. The bound enzymes were not solubilized by 8 M LiCl or neutral or anionic detergents; they were readily detached from walls by a cationic detergent or by autodigestion. Once detached, the enzymes did not reassociate with wall to give salt-resistant complexes. Six enzymes hydrolyzing wall polymers were bound to the envelope, and the same activities were also detected among soluble proteins in the cytoplasmic fraction. It is suggested that cytoplasmic vesicles, containing autolysins, are inserted into or trapped by newly formed wall in the growing hypha; these constitute the wall-bound autolysin fraction. Starvation for a carbon source derepressed the synthesis of five out of the six autolysins, and the amounts of both soluble and wall-bound activities increased by one to two orders of magnitude.


Copy numbers of coexisting plasmids in Escherichia coli K-12. A strain of Escherichia coli K-12 carrying eight compatible and distinguishable plasmids was constructed. The amounts of plasmid DNA (measured as supercoiled molecules) per chromosome in this strain was about equal to the sum of the plasmid DNAs, extracted under controlled conditions, from strains each carrying one of the eight plasmids. Analysis of these DNA preparations showed that each plasmid in the multiplasmid strain was present in the same proportion per chromosome as in the single-plasmid strains. Also the level of phenotypic expression of each plasmid in the multiplasmid strain was the same as in the single-plasmid strains. Each plasmid, therefore, appears to control its own copy number irrespective of the presence of other compatible plasmids.


Involvement of the relA gene product and feedback inhibition in the regulation of DUP-N-acetylmuramyl-peptide synthesis in Escherichia coli. The regulation of uridine diphosphate-N-acetylmuramyl-peptide (UDP-MurNAc-peptide) synthesis was studied by labeling Escherichia coli strains auxotrophic for lysine and diaminopimelate with [3H]diaminopimelate for 15 min under various conditions. The amounts of [3H]diaminopimelate incorporated into UDP-MurNAc-tripeptide and -pentapeptide by a stringent (rel+) strain were the same in the presence or absence of lysine. Chloramphenicol-treated rel+ cells showed a 2.8-fold increase in labeled UDP-MurNAc-pentapeptide. An isogenic relaxed (relA) strain deprived of lysine showed a 2.7-fold increase in UDP-MurNAc-pentapeptide. Thus, UDP-MurNAc-pentapeptide synthesis is regulated by the relA gene. D-Cycloserine treatment of rel+ and relA strains caused a depletion of intracellular UDP-MurNAc-pentapeptide. Labeled UDP-MurNAc-tripeptide accumulated in D-cycloserine-treated cells of the rel+ and relA strains, suggesting that UDP-MurNAc-pentapeptide is a feedback inhibitor of UDP-MurNAc-peptide synthesis. In lysine-deprived cells, D-cycloserine treatment caused 41- and 71-fold accumulations of UDP-MurNAc-tripeptide in rel+ and relA strains, respectively. A 124-fold increase in UDP-MurNAc-tripeptide occurred in lysine-deprived rel+ cells treated with both chloramphenicol and D-cycloserine. These results indicate that both the relA gene product and feedback inhibition are involved in regulating UDP-MurNAc-peptide synthesis during amino acid deprivation.


Relationship of large and small invertases in Saccharomyces: mutant selectively deficient in small invertase. A mutant strain of Saccharomyces cerevisiae (D10-ER1) has been isolated after a two-step mutagenesis of strain 4059-358D (SUC 1) using ethyl methane sulfonate. Cells of this new strain produced a level of total invertase equaling that of 4059 but contained only trace amounts of the small, internal, aglycan form of the enzyme (less than 0.1% of total in D10-ER1 compared with 6% in 4059). When D10.ER1 was crossed with an invertase-hyperproducing strain dgr3 (SUC3), progeny were isolated (HZ400-5A and HZ400-2C) in which levels of total invertase had at least quadrupled. The percentage of small invertase, however, remained insignificant. Levels of small invertase in strain HZ400-5A were determined by affinity chromatography on conconavalin A-Sepharose, gel permeation chromatography, and isopycnic centrifugation in CsCl. The large invertase of the SUC1 yeasts described here was found to contain a form apparently greater in size than the large invertase of the SUC2 strain FH4C; this probably reflects a higher content of carbohydrate. The overall results of this study do not support a direct structural relationship between large and small invertases. The implications on invertase biosynthesis and structure are discussed.


Effect of tsl (thermosensitive suppressor of lex) mutation on postreplication repair in Escherichia coli K-12. Cells of Escherichia coli K-12 carrying lexA or recA mutations are more sensitive to UV radiation than corresponding wild-type cells and are defective in postreplication repair. Supressor mutations (tsl) have been described previously which increase the UV resistance of lexA uvr+, lexA uvrA, and recAI uvr+ strains, but not the resistance of recA1 uvrA strains. We have studied the effect of the tsl-1 mutation on postreplication repair and find that the enhanced survival conferred by this mutation is correlated with an increased capacity for postreplication repair.


Enzymes producing 4-thiouridine in Escherichia coli tRNA: approximate chromosomal locations of the genes and enzyme activities in a 4-thiouridine-deficient mutant. A previously described mutant of Escherichia coli which lacks 4-thiouridine in its tRNA was here shown to be deficient in factor A, one of the two proteins responsible for this thiolation of uridine. Addition of exogenous factor A restored the thiolating ability of extracts prepared from the mutant. The activities of the two thiolation proteins were governed by genes at two widely separated positions on the chromosome, as determined with F-prime merodiploids. The site governing factor A activity lay roughly in the region of the recently reported position of nuv, a gene controlling the production of 4-thiouridine in tRNA.


Intraperiplasmic growth of Bdellovibrio bacteriovorus 109J: solubilization of Escherichia coli peptidoglycan. During penetration of Bdellovibrio bacteriovorus into Escherchia coli, two enzymatic activities, a glycanase and a peptidase, rapidly solubilized some 10 to 15% of the E. coli peptidoglycan. The glycanase activity, which solubilizes peptidoglycan amino sugars, came to a sharp halt with completion of the penetration process. Peptidase activity, which cleaves diaminopimelic acid residues from the peptidoglycan, continued, but at a decreasing rate. By 90 min after bdellovibrio attack, some 30% of the initial E. coli diaminopimelic acid residues were solubilized and present in the culture fluid as free diaminopimelic acid. During bdellovibrio penetration some 25% of the lipopolysaccharide glucosamine was also solubilized by an as yet undefined enzymatic activity that yielded products having molecular weights below 2,000. The solubilization of E. coli lipopolysaccharide glucosamine also terminated at completion of bdellovibrio penetration. At the end of bdellovibrio growth, a second period of rapid solubilization of bdelloplast peptidoglycan began which resulted in lysis of the bdelloplast and complete solubilization of the peptidoglycan amino sugars and diaminopimelic acid. The final lytic enzyme(s) was synthesized just before the time of lysis.


Characterization of a restriction enzyme from Escherichia coli K carrying a mutation in the modification subunit. The restriction enzyme from a restriction and modification-deficient strain of Escherichia coli K mutated in the modification gene (hsdM) has been purified using an in vitro complementation assay with a mutant restriction enzyme from a strain lacking only restriction. The restriction enzyme from the hsdM mutant lacks all of the activities that are associated with the wild type enzyme: binding of unmodified DNA to filters, cleavage, or methylation of unmodified DNA and ATP hydrolysis. It is shown that the enzyme from this hsdM mutant cannot bind S-adenosylmethionine, an allosteric effector in the restriction reaction. In the absence of enzyme activation by S-adenosylmethionine, no binding to unmodified DNA takes place. A comparison with other mutant restriction enzymes allows us to outline the biochemical role of the subunits of the E. coli K restriction endonuclease.


Nucleotide sequence of the L-arabinose regulatory region of Escherichia coli K12. The nucleotide sequence of a 250-base pair segment of L-arabinose operon DNA containing the 150-base pair regulatory region has been determined. This segment includes the promoter for rightward araBAD transcription, pBAD, the promoter for leftward araC transcription, pC, and sites responsible for repression.


Inhibition of primase, the dnaG protein of Escherichia coli by 2'-deoxy-2'-azidocytidine triphosphate. 2'-Deoxy-2'-azidocytidine-5'-triphosphate was investigated as an inhibitor in two reconstructed enzyme systems which catalyze the replication of two viral DNAs. During replication of the duplex replicative form of phiX174 DNA, DNA polymerase III holoenzyme was weakly inhibited and inhibition was reversed by dCTP. A more pronounced inhibition, not reversed by either dCTP or CTP, was observed during replication of the single-stranded DNA of the bacteriophage G4, a close relative of phiX174. This effect depended on the incorporation of 2'-deoxy-2'-azidocytidine-5'-triphosphate by primase (dnaG protein) which synthesizes a 29-residue RNA primer at the unique origin of bacteriophage G4 DNA replication. Extension of the primer strand, terminated by 2'-deoxy-2'-azidocytidine-5'-triphosphate is then severely inhibited. Primase was also inhibited by the 2'-deoxy-2'-azido derivatives of ATP, GTP, and UTP.


The ribosomal protein L24 of Escherichia coli is an assembly protein. Incubation of 50 S subunits with 4.2 M LiCl leads to 4.2c cores and the complementary split protein fraction SP4.2, the latter containing quantitatively L24. L24 was removed from the split fraction by means of CM-cellulose chromatography. Partial and total reconstitution experiments performed with this protein preparation in the absence and presence of L24 demonstrate the crucial role of L24 in the early stage of assembly. However, this protein is dispensable for the subsequent steps of the in vitro assembly. 50 S subunits lacking L24 are fully active in the translation of artificial (poly(U)) and natural (R17 RNA) mRNA, indicating that L24 is not involved in any function of protein synthesis of the mature ribosome. It is therefore a mere assembly protein.


The effects of glucagon, catecholamines, and the calcium ionophore A23187 on the phosphorylation of rat hepatocyte cytosolic proteins. Recent experiments have demonstrated that stimulation of rat hepatocyte alpha-adrenergic receptors alters the activity of enzymes known to be regulated by cycles of phosphorylation and dephosphorylation. These events apparently occur without an increase in the activity of adenosine 3':5'-monophosphate-dependent protein kinase. The present study compared the effects of glucagon and catecholamines on the incorporation of radioactive phosphate into cytosolic proteins obtained from intact rat hepatocytes. Sodium dodecyl sulfate-polyacrylamide slab gel electrophoresis resolved 27 phosphorylated bands in the molecular weight range 220,000 to 29,000. Treatment of the intact hepatocytes with glucagon or cyclic nucleotides increased the phosphorylation of 12 of these bands. Incubation of unlabeled cytoplasmic proteins with the catalytic subunit of protein kinase and [gamma-32P]ATP leads to the phosphorylation of 11 proteins. The molecular weights of these proteins were very similar to those altered by glucagon treatment of intact cells. Stimulation of the alpha-receptor with norepinephrine, epinephrine, or phenylephrine in the presence of 20 micrometer propranolol caused an increase in the phosphorylation of at least 10 of the same 12 phosphorylated bands stimulated by glucagon. The increase in phosphorylation mediated by alpha-receptors was only 50 to 60% of that observed with glucagon  and occurred in the absence of any change in the level of adenosone 3':5'-monophosphate. The effects of alpha-receptor stimulation could be completely antagonized by 20 micrometer ergotamine or 20 micrometer phentolamine. Treatment of the cells with the Ca2+ ionophore A23187 in an attempt to mimic alpha-receptor function increased the phosphorylation of 4 of the phosphoproteins altered by glucagon or catecholamines. The effects of the ionophore depended on the presence of extracellular Ca2+ ion and were similar in magnitude to those of catecholamines. It is concluded that alpha-receptor occupation alters the activity of an adenosin 3':5'-monophosphate-independent protein kinase or phosphatase with a specificity similar to those affected by cyclic nucleotides.


Nucleation of microtubules in vitro by isolated spindle pole bodies of the yeast Saccharomyces cerevisiae. Spindle pole bodies (SPBs) were isolated from the yeast Saccharomyces cerevisiae by an adaptation of the Kleinschmidt monolayer technique. Spheroplasts prepared from the cells were lysed on an air-water interface. Spread preparations were picked up on grids, transferred to experimental test solutions, and prepared for whole-mount electron microscopy. Using purified exogenous tubulin from porcine brain tissue, the isolated SPBs were shown to nucleate the assembly of microtubules in vitro. Microtubule growth was directional and primarily onto the intranuclear face of the SPB. Neither the morphology nor the microtubule-initiating capacity of the SPB was affected by treatment with the enzymes DNase, RNase, or phospholipase although both properties were sensitive to trypsin. Analysis of SPBs at various stages of the cell cycle showed that newly replicated SPBs had the capacity to nucleate microtubules. SPBs isolated from exponentially growing cells initiated a subset of the yeast spindle microtubules equivalent to the number of pole-to-pole microtubules seen in vivo. However, SPBs isolated from cells in stationary phase and therefore arrested in G1 nucleated a number of  microtubules equal to the total chromosomal and pole-to-pole tubules in the yeast spindle. This may mean that in G1-arrested cells, the SPB is associated with microtubule attachment sites of the yeast chromatin.


Immunological characterization of a major transformation-sensitive fibroblast cell surface glycoprotein. Localization, redistribution, and role in cell shape. The major cell surface glycoprotein of chick embryo fibroblasts, cellular fibronectin (formerly known as CSP or LETS protein), was purified and used to produce monospecific antisera. After affinity purification, the anti-fibronectin was used to investigate fibronectin's localization, its transfer from intracellular to extracellular pools, its antibody-induced redistribution on the cell surface, and its role in cell shape. Anti-fibronectin localizes to extracellular fibrils located under and between sparse cells, and to a dense matrix that surrounds confluent cells. Cellular fibronectin is also present in granular intracytoplasmic structures containing newly synthesized fibronectin before secretion. This intracellular staining disappears 2 h after treatment with cycloheximide or puromycin, and returns after removal of these protein synthesis inhibitors. In pulse-chase experiments using cycloheximide, fibronectin was sequentially transferred from the intracellular to the fibrillar extracellular forms. Transformation of chick fibroblasts results in decreases in both extracellular and intracellular fibronectin, and in altered cell shape. Treatment of untransformed chick fibroblasts with anti-fibronectin results in rapid (30 min) alteration to a rounder cell shape resembling that of many transformed cells. These rapid shape changes are followed by a slow, antibody-induced redistribution of fibronectin to supranuclear caplike structures. This "capping" is inhibited by metabolic inhibitors. Reconstitution of cell surface fibronectin onto transformed cells restores a more normal fibroblastic phenotype. The reconstituted fibronectin on these cells organizes into fibrillar patterns similar to those of untransformed cells. As with untransformed cells, treatment of these reconstituted cells with anti-fibronectin also results in cell rounding and "capping" of fibronectin.


Method for rapid detection of group B streptococci by coagglutination. A quick and reliable technique for the identification of group B streptococci has been developed. The method requires no elaborate equipment or expensive reagents and can be used to detect the group B organisms in mixed broth cultures or to identify suspect colonies selected from agar plates. The method is a coagglutination technique in which 1 drop of specifically sensitized protein A-containing Staphylococcus aureus is mixed with 1 drop of supernatant of an actively growing culture. The soluble group-specific carbohydrate substance of the group B streptococci reacts with the staph particles to produce agglutination that is macroscopically readable. One colony of group B streptococci taken from an agar plate and inoculated into Todd-Hewitt broth will give a positive reaction within 6 h of incubation; with a larger inoculum, the positive reaction occurs within a shorter period. The method was applied for detection of group B streptococci in mixed broth cultures. In laboratory studies involving random mixtures of organisms, 59.3% of positive cultures were detected within the first 8 h of incubation, and 71.7% were found within 24 h. In clinical studies with mixed broth cultures grown directly from vaginal swabs, 78.6% of the positive cultures were detected within the first 8 h of incubation, and 92.9% were found within 24 h.


Detection of antibodies to legionnaires disease organism by microagglutination and micro-enzyme-linked immunosorbent assay tests. Microagglutination and micro-enzyme-linked immunosorbent assay (ELISA) tests with easily prepared, safe, heat-killed antigens for detecting antibodies to the legionnaires disease organism have been developed. A safranin-stained whole-cell antigen is used in the microagglutination test, and a simply prepared soluble antigen is used in the micro-ELISA tests. The microagglutination test detected elevated titers in 97.2% of the sera from patients with legionnaires disease. Three variations of the micro-ELISA test with anti-human immunoglobulin G, immunoglobulin M, and Fab peroxidase-labeled conjugates revealed elevated titers with 74.3, 82.9, and 88.6% of the sera, respectively. The microagglutination and the micro-ELISA tests used in combination detected 100% of the elevated titers.


Evaluation of a simplified Guizotia abyssinica seed medium for differentiation of Cryptococcus neoformans. A simplified Guizotia abyssinica seed medium, eliminating glucose, creatinine, and phosphate, was evaluated for the isolation and presumptive identification of Cryptococcus neoformans. Of 80 isolates of C. neoformans tested, 69 (86%) developed the characteristic brown pigment within 12 h on this medium as against only 5 isolates (6%) on the complete medium. In primary cultures of experimentally seeded specimens of sputum, bronchial aspirate, soil, and pigeon excreta, C. neoformans was recognizable within 3 to 5 days on the modified medium in contrast to the 3 to 8 days required on the complete medium. The results demonstrated that the simplified G. abyssinica seed agar, with or without diphenyl, is superior to the complete medium  for the rapid development of the brown pigment by C. neoformans.


Clinical evaluation of the Uni-Yeast-Tek system for rapid presumptive identification of medically important yeasts. The results of over 400 tests for identification of clinical yeast isolates as to species using the Uni-Yeast-Tek (UYT) system in comparison with a more conventional system are reported. The conventional system utilized a total of 23 individual tests, including both fermentation and assimilation tests, whereas the UYT system included only 11 separate tests. In the initial phase of the study, coded unknown isolates were evaluated by each of two technologists using both methods independently. After this initial evaluation, the two methods were used in parallel for routine testing of yeast isolates as they were obtained from clinical specimens. A further evaluation of the UYT system was carried out by retrospectively analyzing the species reported from a clinical mycology laboratory during two separate time periods in which different approaches to yeast identification were employed. A total of 92% of the isolates tested with the UYT system were correctly reported within 72 h, 96% were correctly named after 1 week of incubation, and 97% were correctly reported after 2 weeks of incubation of UYT plates at 30 degrees C when results of the two phases of the study were analyzed together. With the conventional system, 88% of the isolates were correctly reported at 72 h, 96% at 1 week, and 98% after 2 weeks of incubation of biochemical tests. Retrospective analysis of laboratory records revealed no major changes in species reported after adoption of the UYT system for routine testing of clinical isolates. The data presented in this report suggest that the UYT system can be expected to yield rapid presumptive identification of clinical yeast isolates with reasonable confidence when certain minor limitations that are discussed in the text are taken into account.


Direct-plate serological grouping of beta-hemolytic streptococci from primary isolation plates with the Phadebact streptococcus test. The grouping of beta-hemolytic streptococcal isolates by a new direct-plate procedure employing Phadebact Streptococcus Test reagents was compared with the results obtained with the 4- and 24-h Phadebact grouping procedure and with the Lancefield grouping obtained with a capillary precipitin test. The new procedure employed a modification of the Phadebact procedure that permitted the grouping of streptococci on glass slides with a minimum of five primary isolated colonies. When only five to eight colonies were available for direct testing with each Phadebact reagent, coagglutination was better manifested when the colonies were disaggregated on a glass slide in a loopful of Tween 80 solution. Further enhancement of the coagglutination reaction was effected when the respective Phadebact reagents were employed in relatively small volumes. The direct-plate procedure permitted the correct identification of 127 out of 129 betahemolytic isolates. The 4-h method correctly identified 192 of the 200 streptococci tested. All of the 200 isolates tested by the 24-h procedure and the Lancefield grouping were correctly identified. The direct-plate Phadebact procedure affords the clinical microbiologist a rapid and reliable means of identifying groups A, B, C, and G beta-hemolytic streptococci. When sufficient numbers of primary colonies are not available for the direct procedure, the 4- or 24-h procedures may be employed.


Serological reactions of the genus Peptostreptococcus. White male New Zealand rabbits were immunized with soluble antigen preparations (SP) of the following gram-positive anaerobic cocci: Peptostreptococcus anaerobius ATCC 27337 and VPI 5737; P. micros VPI 2618; Streptococcus morbillorum ATCC 27527; P. parvulus VPI 5229; and P. productus ATCC 27340. SP were reacted with homologous and heterologous rabbit antisera in immunodiffusion, immunoelectrophoresis, indirect fluorescent-antibody, and tanned-cell passive hemagglutination tests. Even though each antiserum reacted strongly with its homologous SP, no interspecies reactivity was observed except between P. productus antisera and P. parvulus SP by the passive hemagglutination test. Antisera prepared to both strains of P. anaerobius reacted with the other strain in all serological tests.


Counterimmunoelectrophoresis for detection of microbial antigens: increased sensitivity with dextran-containing gels. The addition of 4% dextran with a mean molecular weight of 70,000 to counterimmunoelectrophoresis gels enhanced the clarity of precipitin lines and increased the sensitivity of the procedure two- to fourfold with a variety of microbial antigens.


Development of a template for use in immunofluorescent identification of mycoplasmas. A template for securing blocks of agar onto glass slides is described. Use of the template allows efficient handling as well as thorough washing of agar blocks during the immunofluorescence staining process.


Microwave oven for melting laboratory media. A microwave oven was used to melt microbiological media, resulting in a substantial reduction of heat generation and considerable savings in time.


Glucose-stimulated 45Calcium efflux from isolated rat pancreatic islets. Kinetics of (45)Ca efflux and insulin release were studied in collagenase-isolated rat islets during 2-h perifusions with calcium-depleted (0.05 mM) bicarbonate-phosphate buffer containing 2.2 mM glucose. Addition of glucose (16.7 mM) suppressed (45)Ca efflux by 30%. Removal of glucose caused an "off response" of insulin release. The perifusion of a normal concentration of Ca (2.3 mM) greatly stimulated (45)Ca efflux, indicating Ca <--> (45)Ca exchange. When Ca and glucose were superimposed, the effects on (45)Ca efflux and insulin release depended upon the order of presentation of the stimuli: when Ca was added to an ongoing 16.7-mM glucose perifusion, biphasic patterns of (45)Ca and insulin release were seen; when glucose was superimposed on a Ca perifusion, an inhibition of the Ca-stimulated (45)Ca efflux occurred, and a reduced but clearly biphasic insulin response was seen. The subsequent insulin off response after with-drawal of the glucose was also reduced. Mathematical "peeling" of (45)Ca efflux curves from unstimulated islets suggests that there are at least two, and probably three, different intracellular Ca compartments (not including the extracellular sucrose space). At the beginning of perifusion, these three compartments (I, II, III) contain 25, 56, and 19% of the intracellular (45)Ca, and their rates of efflux are 6.7, 1.2, and 0.1%/min, respectively. Glucose appears to suppress efflux from the largest compartment (II); Ca appears to exchange with (45)Ca from a more inert compartment (III). The relationship between insulin and (45)Ca release is not stoichiometric.


Antibodies to histones in drug-induced and idiopathic lupus erythematosus. When tissue sections are extracted with 0.1 N HCl, cellular nuclear proteins, including histones, are removed but nuclear DNA is retained. Histones can be reconstituted back to nuclear DNA in acid-extracted tissue sections so that the resulting nuclear substrate is composed only of DNA and histones and does not contain acidic nuclear protein antigens. The resulting DNA-histone tissue substrate can be used in the immunofluorescent method for specific detention of antibodies to histones. Sera from 23 patients with drug-induced lupus erythematosus (procainamide 19, isoniazid 2, nitrofurantoin 2) and 20 patients with idiopathic (not drug-induced) systemic lupus erythematosus (SLE) were studied. All 23 patients with drug-induced lupus erythematosus (LE) lost nuclear staining on acid-extracted sections. In contrast, only 12 of 20 with idiopathic SLE lost nuclear staining on acid-extracted tissues, and in the remaining 8, there was no significant fall in titer. In the drug-induced LE group, loss of nuclear staining was due to the absence of histones on the substrate because with histone-reconstituted sections, 22 of 23 again became positive for nuclear staining at titers equal to or at one doubling dilution below titers on unextracted tissues. In contrast, of the 12 idiopathic SLE sera which lost nuclear staining, only 5 regained nuclear staining on histone-reconstituted tissue sections. In idiopathic SLE, antinuclear antibodies are heterogeneous in specificities and may consist of antibodies to native DNA, histones, or nonhistone proteins. In contrast, antinuclear antibodies in drug-induced LE are less heterogeneous and mainly consist of antibodies to histones.


Renin response to stimulation of cardiopulmonary mechanoreceptors in man. Plasma renin activity (renin) and hemodynamic response to venous pooling of blood in legs were studied in 24 healthy volunteers and four patients who after bilateral nephrectomy received a functioning renal transplant. Blood pressure cuffs were placed around subjects' thighs and inflated at a pressure 5 mm Hg below the individuals' diastolic pressures. 30 min after thigh cuff inflation, renin significantly increased in all volunteers (mean = 125%). Inflation of cuffs induced a decrease of right atrial pressure, cardiopulmonary blood volume, and cardiac output, but there were no changes in the extra-arterial systolic and diatoloic pressure or in the pressure amplitude. After cuffs were deflated, renin and hemodynamic parameters returned toward normal. In nine volunteers in whom thigh cuff inflation initially elicited renin increases, subsequent intravenous propranolol (0.25 mg/kg) abolished the response to repeated cuff inflation. The renin increase to thigh cuff inflation was absent or suppressed in four patients with a recently transplanted denervated kidney. It is concluded that thigh cuff inflation elicited a reflex-mediated renin increase, and that the reflex stemmed from stimulation of cardiopulmonary mechanoreceptors.


Effects of cue-controlled relaxation, a placebo treatment, and no treatment on changes in self-reported test anxiety among college students. Twenty-eight test-anxious Ss received either "cue-controlled relaxation therapy," an inert placebo treatment, or no treatment. Once before and twice after the experimental treatment period each S responded to three test-anxiety scales and to one measure of public-speaking anxiety. Once after the treatment, Ss rated the credibility of the procedure to which they had been exposed. Pre- and posttreatment scores on the measure of (untreated) speech anxiety and posttreatment ratings of credibility both showed that the cued relaxation and placebo procedures exerted equivalent experimental demand for improvement. Scores on all three test-anxiety measures showed that, under these conditions of equivalent demand,the placebo procedure reduced self-reports of test anxiety at least as much as did cue-controlled relaxation. Implications are discussed.


Factorial relationships of measures of perceived level of psychopathology and sociometric choice. Studied the relationship of diagnosis, age, length of hospitalization, and various measures of perceived level of psychopathology, perceived appropriateness of hospitalization, and sociometric choice among 39 adolescent psychiatric inpatients. Factor analysis revealed two factors, one that was apparently a measure of perceived level of psychopathology and another that was apparently a measure of sociometric choice. Perceptions of an individual's level of psychopathology appear to be shared across staff and patient boundaries. Appropriateness of hospitalization had a strong positive relationship to perceived level of psychopathology and a weak negative relationship to sociometric standing. The relative lack of relationship between sociometric standing and perceived level of psychopathology is discussed in light of past research and methodological design issues.


The pain apperception test: an investigation of concurrent validity. Gave 55 chronic pain patients the Pain Apperception Test (PAT), the MMPI, the Health Index Index, the Whitely Index, the Pain Estimate, and the Tourniquet Test in an investigation of the concurrent validity of the PAT. The results did not support strongly the concurrent validity of the PAT, given the small number of significant correlations obtained. However, several consistent patterns did emerge: The PAT appeared to be correlated positively with the Alcoholism and Social Introversion scales of the MMPI and with the Invalidism scale of the Health Index. The PAT appeared to be correlated negatively with the Psychopathic Deviate scale of the MMPI. The latter results are discussed with respect to the psychodynamics of chronic pain patients. Further research is suggested, especially with regard to direct pain measures and predictive validity.


Retrospective identification of Acanthamoeba culbertsoni in a case of amoebic meningoencephalitis. Acanthamoeba culbertsoni was identified retrospectively in a case of amoebic meningoencephalitis, previously reported by Jager and Stamm (Lancet, 2, 1343, 1972). This is the second report of this species causing secondary infection in man. Positive results were obtained only with anti-A. culbertsoni sera when the brain sections were stained by the indirect immunofluorescence antibody test with various antisera produced against different Acanthamoeba species. Antiserum raised against purified plasma membranes of A. culbertsoni showed once more its highly specific diagnostic value.


A stereotaxic atlas of the brain of the three-day-old domestic chick. Preparation of a stereotaxic atlas of the brain of the 3-day-old domestic chick was prompted by the widespread use of chicks in behavioral and pharmacological research. The atlas is comprised of 26 frontal plates extending from the frontal pole to the level of the hypoglossal nucleus, and one sagittal plate at lateral 0.4 mm. Marking lesions were used to ascertain coordinates, the brains were embedded in albumin and sections cut at 36 mmu. The brain sections were stained with cresyl violet for nuclei and with hematoxylin for fibers. Unstained brain sections were used as a guide in drawing the plates to correct for shrinkage and distortion. The use of the atlas for research purposes for one year, plus test lesions targeted for specific brain structures, indicates the atlas is accurate to within 0.5 mm.


Characterization of the monoaminergic innervation of immature rat neocortex: a histofluorescence analysis. In the neocortex of 6-day-old rat, abundant axon terminals which exhibit specific catecholamine fluorescence are found in all regions and throughout all cortical layers. The overall density of axons in 6-day-old cortex is similar to the density in the adult cortex. In immature cortex, there are two distinct fluorescent plexuses, both presumably noradrenergic, one in the molecular layer and another in the lower half of the cortex. The superficial plexus is composed primarily of horizontal fibers, and the deep plexus of a dense feltwork of obliquely oriented fibers suggestive of a terminal field. The cortical plate itself is traversed by a few vertical processes. Following lesions of the midbrain tegmentum no fluorescent axons are seen in cortex, providing evidence that the fluorescent axons in cortex arise from brain stem neurons. The deep and superficial plexuses can be differentially visualized depending on the histochemical techniques employed and on pharmacological treatment, such as loading with a monoamine congener. Both deep and superficial axons are shown to contain endogenous catecholamines but those fibers in the deep plexus are filled to far less than their maximum capacity. The pharmaco-histochemical differences between axons in the two plexuses suggest that there may exist two distinct catecholaminergic projections to lateral neocortex. The demonstration of an extensive cortical monoamine innervation early in ontogeny supports the possibility that monoamine neurons play an important role in information processing and/or developmental interactions in the immature brain.


Effect of sublingual pentrinitrol on exercise tolerance in angina pectoris. The effects of 10 mg sublingual pentrinitrol and placebo on the exercise tolerance of 20 patients with angina pectoris were compared. The ten patients who received pentrinitrol had increases in exercise times and metabolic oxygen equivalents (METs) 15 and 60 minutes after administration, whereas no change occurred in ten comparable patients receiving placebo. Pentrinitrol produced lower preexercise systolic blood pressures and systolic blood pressures at point of angina. An increase in heart rate offset the effect of the reduced systolic blood pressure on double product (systolic blood pressure times heart rate). Sublingual pentrinitrol is an effective antianginal drug for at least 60 minutes.


Hypnotic activity of diphenhydramine, methapyrilene, and placebo. In a double-blind controlled study, an oral dose of diphenhydramine hydrochloride (12.5, 25, or 50 mg), methapyrilene fumarate (36, 72, or 144 mg), or placebo was administered to 1295 post-partum patients if they complained of, or anticipated, a sleep problem. Hypnotic activity was assessed clinically by subjective and objective techniques. Methapyrilene and diphenhydramine, at all doses, were found to be effective hypnotics in comparison to placebo, based on sleep latency, sleep duration, awakening in the night, global evaluation, and morning alertness. Increasing the dose of these drugs, in the range studied, produced a minimal increase in effectiveness.


Nasal CPAP employing a jet device for creating positive pressure. A nasal CPAP system consisting of a jet device (weight 5 g) connected with a binasal nose-piece is described. A positive airway pressure is obtained with the system applied to newborn infants provided a certain magnitude of air is brought to the jet device, and a resistance, presented by the airways, exists against the flow of air. As the system requires neither a manometer nor an excess pressure safety valve, pressures in the nasopharynx were recorded at various jet flows to illustrate the flow/pressure relationship. The CPAP can easily be etablished at definite pressures by simple adjustment of the jet flow, using this flow/pressure curve. The system appeared efficient for the treatment of apnoea, apparently due to a special attribute of the jet device. Oesophageal pressures were measured at various gas flows and at various stages of RDS, and in children with healthy lungs. The pressure appeared high with increased density of the lungs, but constant in healthy lungs despite various gas flows. Extreme pressure excursion were recorded during crying and increased muscular tone. This may explain the high frequency of pneumothorax in RDS.


Nasal CPAP treatment of the respiratory distress syndrome: a prospective investigation of 10 new born infants. Ten newborn infants with the idiopathic respiratory distress syndrome (IRDS) or foetal aspiration (FA) were treated with a simplified nasal CPAP system (continuous positive airway pressure). The system consists of a Hudson binasal cannula and a Benveniste pediatric jet device (weight 5 g). The desired airway pressures are obtained by adjustment of the jet flow in accordance with the naso-pharyngeal flow/pressure relationship. The system was found to be effective in the treatment of 9 of the 10 children, as demonstrated by improved blood gas values, and improved respiration; the latter was also evident in infants with pronounced apnea. In one child pneumothorax, occuring after about 24 hours of successful CPAP treatment, necessitated respirator treatment. The system is quick and simple to use, and permits feeding, lung physiotherapy and routine nursing of the child during treatment.