Gm and Km(Inv) frequencies in two Roumanian populations.

Serum samples from 170 unrelated individuals from the Suceava District of Roumania and from 199 unrelated individuals from Bucharest, Roumania were tested fro Gm(1,2,3,5,6,13,14,17,21) and Km(1)[Inv(1)]. Selected samples were also tested for Gm(15) and Gm(16). The frequencies of the three common Caucasoid haplotypes, Gm3,5,13,14, Gm1,17,21, and Gm1,2,17,21 in these two populations were found to be similar to those in neighboring Slavic states and Hungary. Racial admixture was evidenced by the presence of the Gm1,13,15,16,17 and Gm1,3,5,13,14 haplotypes, which are primarily Mongoloid, and the Gm1,5,13,14,17 haplotype which is primarily Negroid. Comparisons of these data with those from earlier studies of populations from Central Europe indicate that the frequency of the Gm3,5,13,14 haplotype within this region is high and essentially uniform. Published data for several blood group systems also indicate essentially uniform distributions of frequencies in this region. It is suggested that this region may be the center of a cline that radiates from it.


Characterization of residual hexosaminidase activity in Sandhoff's disease using man-Chinese hamster cell hybrids.

To obtain information about the nature of the residual hexosaminidase activity in Sandhoff's disease, hybrid cell lines between fibroblasts from a patient with Sandhoff's disease and Chinese hamster cells were isolated. In these hybrid cell lines, a heteropolymeric isoenzyme was detected that is composed of human alpha- and Chinese hamster hexosaminidase subunits. Due to the electrophoretic and immunological behavior of the heteropolymeric molecules in interspecies hybrids with normal fibroblasts and with cells from a patient with Sandhoff's disease, we conclude that Sandhoff cells contain an alpha-subunit of hexosaminidase with normal characteristics.


Beta lactamase resistance of newer cephalosporins and antimicrobial effectiveness against gram-negative bacilli.

Three newer cephalosporins (cefamandole, cefoxitin and cefazaflur) were investigated, in comparison with three older agents (cephalothin, cephaloridine and cefazolin) to determine their stability to beta-lactamases of gram-negative bacilli, and to correlate this with their antibacterial activity. Nine of the 17 bacterial strains employed produced broadspectrum beta-lactamases; the remaining eight produced cephalosporinases. The cephalosporins were highly active against bacteria producing broad-spectrum beta-lactamases; they were less active against organisms producing cephalosporinases. All of the cephalosporinase-producing strains were resistant to cephalothin anc cephaloridine. With the other cephalosporins the correlation between hydrolysis by cephalosporinases and resistance of the organisms was poor. Four to eight cephalosporinase-producing strains were resistant to cefoxitin, which was completely resistant to hydrolysis by the beta-lactamases. Cefozolin, cefamandole and cefazaflur inhibited several of these strains in spite of destruction by the beta-lactamase. Several cephalosporins need to be used in antimicrobial susceptibility testing of gram-negative bacilli.


Protective isolation and antimicrobial decontamination in patients with high susceptibility to infection. A prospective cooperative study of gnotobiotic care in acute leukemia patients. II. Organizational and statistical concept.

The organizational and statistical concept of the cooperative randomized controlled clinical trial on gnotobiotic care and its effect in acute leukamia patients is described. The purpose is the clarification of the concept of the study and the comparison with other studies on the same subject. It is concluded that the value of the results and of the decisions based on them depend mainly on the statistical design, the protocol, and the performance of a study. The performance of a controlled study depends on its organizational details, its monitoring, documentation, and appropriate biometric evaluation. In this particular study it proved fruitful that a statistician was engaded in the evaluation and also in developing the protocol and in carrying out the study.


Experimental evaluation of HR756, a new cephalosporin derivative: pre-clinical study.

HR 756 is a new cephalosporin derivative suitable for parenteral use. The compound possesses an unusally broad spectrum of antibacterial activity especially against gram-negative bacteria. Besides Escherichia coli, Salmonella, Klebsiella, indole-negative Proteae and other species also indol-positive Proteae, Serratia marcescens, Enterobacter and many Pseudomonas aeruginosa strains are inhibited by this compound. HR 756 is stable to most of the beta-lactamases produced by gram-negative organisms. Tests on different infection models also provided evidence of the high efficacy of HR 756 in vivo.


Immunological relationships of human and subhuman primate pregnancy-associated plasma proteins.

(1) Four pregnancy-associated plasma proteins cross-reactive with antibodies to the human pregnancy proteins were detected in several species of pregnant subhuman primates. In the case of the two apes studied (chimpanzee and orangutan), these appeared to be immunologically identical to the human PAPPs (PAPP-A, -B, -C, and HCS). In the old world monkeys analyzed, equivalent partially cross-reactive PAPPs were found; while in the new world squirrel monkey, only faint traces of cross-reactive PAPP-C and HCS were observed with the most sensitive methods used. (2) As in the human, these proteins appeared to be specific for pregnancy, not being detectable in nonpregnant animals, female or male. (3) The pregnant chimpanzee possessed significantly higher concentrations of PAPP-A and PAPP-C than did women at an equivalent stage of pregnancy, while the HCS and PAPP-B levels appeared to be approximately the same. (4) The primate PAPP-C analogues were more complex than human PAPP-C, often revealing multiple gel diffusion patterns with subfractions of differing electrophoretic mobilities. (5) Based on the changes of electrophoretic mobility upon exposure to neuraminidase, the subhuman primate as well as human PAPP-A and PAPP-C appeared to be glycoproteins containing sialic acid. (6) In the chimpanzee and rhesus monkey, as in the human, the levels of PAPP-A, PAPP-C, and HCS were appreciably higher during the third trimester of pregnancy than they were during the second trimester.


Genetic evidence for a temperature-sensitive lesion in the adenovirus 7 region of the PARA genome.

It has previously been shown that the helper adenovirus (Ad) present in the defective Ad 7-SV40 hybrid population is temperature-sensitive (ts) for replication in human cells. This study has shown that the replication of the defective hybrid virus (PARA) in green monkey kidney cells is also restricted at 40.5 percent. Complementation tests between the parental or transcapsidant PARA populations and SV40 or various Ad serotypes revealed that the ts lesion is located in the Ad region of the hybrid genome. Wild-type (wt) Ad type 7 and Ad type 21 were able to complement the replication of PARA while Ad type 31 was unable to do so. Complementation was more efficient after the PARA genome was transcapsidated to the wt isolates of helper Ad than during multiple infections with the parental hybrid population. The SV40 function which complements the replication of human adenoviruses in simian cells is not expressed by PARA at the nonpermissive temperature. However, the ts lesion does not affect the expression of SV40 functions coded for by PARA under other conditions, i.e., complementation of the replication of human adenoviruses in transformed monkey cells, and synthesis of T antigen in lytically infected or transformed cells. Although the exact nature of the mutated Ad 7 gene product is unknown, heat inactivation data suggest that it may be a structural protein.


[Diagnosis and therapy of the zinc deficiency syndrome during total parenteral feeding. Review with report of a case].

A 47-year-old man with ulcerative colitis developed severe zinc deficiency during longterm intravenous feeding. Early symptoms included seborrhoeic lesions in the face and scalp. Later on his general condition deteriorated, and erythema and bullae erupted in the fingers, together with an erosive area with a marked border in the perianeal and scrotal regions. The symptomatology was suggestive of zinc depletion syndrome, a recently recognised acrodermatitis-enteropathica-like disorder in patients receiving longterm total intravenous feeding. A significantly lowered serum zinc level and a prompt beneficial response to intravenous zinc therapy (20 mg Zn2+ daily for one week) substantiate the provisional clinical diagnosis. As zinc depletion is caused by a zinc free parenteral alimentation this serious complication should be avoided by adding zinc to the infusions. Serum zinc determinations should be carried out as a control of the prophylactic zinc supply. Possibly, zinc plays a role in the symptomatology of various disorders with skin and bowel symptoms.


[Regression of xanthomas in endogenous hypertriglyceridemia under low carbohydrate diet].

Report on two patients with endogenous hypertriglyceridemia. In both patients normal serum lipid values were reached in a comparatively short time under a diet with reduced carbohydrates and calories. In one diabetic patient who needed insulin at the beginning of the treatment the disease could be controlled by dietary measures alone after a few days. Different biological half-life periods of the various serum lipid fractions explain why, under a reducing diet, the rate of decrease of lipids is variable according to the respective component. Thus, the ratio of triglycerides and cholesterol can vary in the same patient within a few days and may change his classification under different types of hyperlipidemia according to Fredrickson. It would appear that in these cases a classification based on etiological considerations is more recommendable.


Cervical esophagostomy to permit extraoral feeding of the horse.

An indwelling stomach tube was placed through a cervical esophagostomy in 5 clinically normal horses and in 3 horses requiring artificial alimentation. In all but one case, surgery was performed on conscious horses following tranquilization and infiltration of the surgical sites with a local anesthetic. Six horses were fed liquid rations through the indwelling tubes, which were left in place from 3 to 26 days. After the tubes were removed, the stomas healed rapidly and with minimal complications. Cervical esophagostomy proved to be a safe, easy, and effective method for the provision of extraoral feeding of the horse.


Measurement of neonatal equine immunoglobulins for assessment of colostral immunoglobulin transfer: comparison of single radial immunodiffusion with the zinc sulfate turbidity test, serum electrophoresis, refractometry for total serum protein, and the sodium sulfite precipitation test.

Four procedures for assessment of adequacy of colostral immunoglobulin (Ig) transfer in foals were evaluated. Results of zinc sulfate turbidity test, serum electrophoresis, total serum protein refractometry, and sodium sulfite precipitation test were compared with immunoglobulin G content determined by single radial immunodiffusion. The zinc sulfate turbidity test gave acceptable results for IgG, except that hemolyzed serum samples gave higher than expected values. A correction factor for hemolyzed serum was found to be useful. Serum electrophoresis was a satisfactory method of estimating IgG content. Total serum protein values may not be a valid basis for estimating IgG content, inasmuch as postsuckling total protein values were found to decrease in some foals in which passive transfer of IgG had been adequate. Sodium sulfite precipitation reactions were too unpredictable to be of value for determination of neonatal IgG concentration.


Listeriosis in an Arabian foal with combined immunodeficiency.

A 1-month-old Arabian foal with signs of central nervous system disease was found to have combined (B- and T-lymphocyte) immunodeficiency. The foal died in spite of intensive antibiotic therapy. At necropsy, generalized lymphoid hypoplasia and acute necrotizing and granulomatous inflammation of the brain, heart, and adrenal glands were found. In addition, there were spinal meningitis and focal hepatic necrosis. Listeria monocytogenes was isolated on primary culture from the brain.


Pholipomycin, a new member of phosphoglycolipid antibiotics. III. Biological properties.

Pholipomycin, a new member of the phosphoglycolipid antibiotics, was primarily active against gram-positive bacteria including clinically isolated resistant bacteria. It differed from other members of the antibiotic family as it also demonstrated activity against gram-negative bacteria. Pholipomycin protected mice from infection with Staphylococcus aureus and was nontoxic to mice having an LD50 (i.v.) of 600 mg/kg. Besides possessing anti-microbiol activity, pholipomycin, when administered orally, appeared to promote growth in chickens and pigs.


Experiences in the search for anti-inflammatory agents of microbial origin.

Whole shaken cultures of 20 random, unidentified actinomycetes were extracted with n-butanol at pH 4.5, 7.0 and 8.5, respectively. Residues of butanol-extractable materials (BXM) were reconstituted (100X) in buffers and freeze-dried. BXM were surprisingly well tolerated in animals and were screened against influenza A viral pneumonia in mice. One culture yielded BXM-80 which suppressed both chemical (LPS) and viral (NDV) pneumonia in mice as well as inhibited rat foot pad edema induced by carrageenin. Aspirin, Butazolidin, hydrocortisone, indomethacin, and prednisolone, which are known to inhibit carrageenin-induced rat foot pad edema were tested against chemical (LPS) and viral (NDV) pneumonia in mice. Only hydrocortisone and prednisolone suppressed LPS pneumonia. All of these 5 compounds failed to inhibit NDV pneumonia. Microbial products are suggested as a source for new and unique anti-inflammatory agents.


Possible beta-lactamase activities detectable in infective clinical specimens.

Biological beta-lactam antibiotic-inactivating activities were detected in bacteriuria and suppurating pleural fluids. Clinical specimens were sterilized with membrane filters and the amounts of ampicillin and/or cephalothin which were being inactivated by 1 ml of each filtrate were determined. In general, filtrates which originally yielded Klebsiella sp. tended to show activity against ampicillin; whereas those yielding Enterobacter sp. and Pseudomonas aeruginosa showed activity against cephalothin.


Development of social behavior in an adult total isolate rhesus monkey (Macaca mulatta).

A 12-year-old female total isolate rhesus monkey was pretested with age mates and subsequently housed for 20 weeks with an infant "therapist" monkey. Daily observations during that period revealed a 24-fold increase in the probability of social behavior. Self-directed behaviors also increased significantly. Disturbance behaviors (self-slapping, self-biting, bizarre limb movements, etc.) remained unchanged. Although problems obviously exist in cross-species generalization, and are here considered, these results emphasize the importance of early therapeutic intervention as well as the need for a more comprehensive approach to both social and disturbance behaviors if the treatment of adults is to be as successful as the treatment of immature isolate subjects.


Biogenesis of endoplasmic reticulum membrane in rat liver cells. I. Intracellular sites of synthesis of cytochrome b5 and NADPH-cytochrome c reductase.

The sites of synthesis of microsomal membrane proteins, NADPH-cytochrome c reductase and cytochrome b5, were investigated by three methods; the in vitro synthesis of these proteins by isolated rough microsomes, the immunoprecipitation of polyribosomes carrying their nascent peptides, and the immunoprecipitation of in vivo-labeled nascent peptides. The in vitro incorporation experiment confirmed that the synthesis of these microsomal proteins was carried out by the bound polyribosomes of rough microsomes. When free and bound polyribosomes were separately examined by the other two methods, we found that NADPH-cytochrome c reductase was synthesized by both classes of polyribosomes whereas cytochrome b5 was synthesized only by bound polyribosomes.


Biogenesis of endoplasmic reticulum membrane in rat liver cells. II. Discharge of the nascent peptides of NADPH-cytochrome c reductase and cytochrome b5 on the cytoplasmic side of the endoplasmic reticulum membrane.

The direction of discharge of the nascent peptides of NADPH-cytochrome c reductase and cytochrome b5 from bound polyribosomes of rough microsomes was investigated in order to elucidate the mechanism of separation of these membrane proteins from secretory proteins, which are also synthesized by the same class of ribosomes of rough endoplasmic reticulum. The nascent peptides of NADPH-cytochrome c reductase and cytochrome b5 in intact rough microsomes were accessible to externally added 125I-Fab's against these proteins, and were susceptible to trypsin digestion, whereas the nascent peptides of serum albumin were not. The nascent peptides of these two microsomal proteins were released into the cytoplasm by puromycin treatment of intact rough microsomes, while the nascent peptides of serum albumin were retained in the microsomal lumen. These observations suggest that the nascent peptides of microsomal proteins, which are present on the cytoplasmic surface of the endoplasmic reticulum membrane, are exposed on the surface of microsomal vesicles, while those of secretory proteins are enclosed inside the vesicles. Therefore, the topographical separation of microsomal membrane proteins from secretory proteins is accomplished at the step of their synthesis by the bound polyribosomes of rough endoplasmic reticulum.


Studies on salivary gland ribonucleases. II. Purification of ribonucleases from bovine submaxillary gland and the effects of polyamines on their activities.

Four alkaline ribonucleases [EC 3.1.4.22] were purified 2,050- to 3,460-fold from bovine submaxillary gland by repeated CM-Sephadex C-25 chromatography and Sephadex G-50 gel filtration, with a total recovery of about 13%. These were designated as RNase BS1, BS2, BS3, and BS4, based on their order of elution from a CM-Sephadex C-25 column. The molecular weights of these enzymes were estimated by gel filtration to be 19,000, 17,500, 17,000, and 12,000, respectively. These enzymes are very similar to RNase A in that they are inhibited by heparin, show preferential hydrolysis of C5'-O-P linkages adjacent to a cytosine nucleotide rather than a uracil nucleotide, and in their antigenic properties. Spermine was found to stimulate the activities of these enzymes; the degree of stimulation was in the order RNase BS4 greater than BS3 greater than BS2 greater than BS1. The stimulation by spermine is due to the increased cleavage of C5'-O-P linkages adjacent to cytosine nucleotide. The reason for the differences in the degree of spermine stimulation of these enzymes is discussed.


A cell wall proteo-heteroglycan from Piricularia oryzae: further studies of the structure.

The carbohydrate part of a proteo-heteroglycan from Piricularia oryzae was further studied by chemical and immunological methods. Acetolysis studies of the proteo-heteroglycan and exo-alpha-D-mannanase resistant core showed a (1 leads to 6) mannan back-bone structure with side chains composed of one to four mannose units, and some of which are terminated by D-glucose or D-galactofuranose. The mode of attachment of the terminal glucose was characterized to be alphaGlc(1 leads to 6)Man by inhibition reaction with oligosaccharides. Rabbit anti-serum formed against P. oryzae cells had three specificities, the first one for alphaGlc(1 leads to 6)alphamannosyl, the second one for alphaMan(1 leads to 3)alphamannosyl, and the last one for alphaGal-f(1 leads to 2)alphamannosyl residues. The most immunodominant side chain structure of the P. oryzae heteroglycan was shown to be alphaGlc(1 leads to 6)alphaMan(1 leads to 2)alphaMan(1 leads to 2)Man.


Structural and immunochemical studies on D-arabino-D-mannans and D-mannans of Mycobacterium tuberculosis and other Mycobacterium species.

Serologically active D-arabino-D-mannas ([alpha]D, +82 degrees approximately 89 degrees; ratio of D-arabinose to D-mannose, 1-2:1) were isolated from the soluble fraction of disintegrated cells of M. tuberculosis, M. smegmatis, and several other Mycobacterium species. These arabinomannans had similar structures, consisting of alpha-(1 leads to 5)-linked D-arabinose residues and alpha-(1 leads to 6)-, and (1 leads to 2)-linked D-mannose residues. Methylation and enzymic degradation studies using Arthrobacter sp. alpha-D-mannosidase and M-2 enzyme (D-arabinan hydrolase) indicated that the arabinomannan of M. tuberculosis Aoyama B possesses short side chains built up from alpha-(1 leads to 2)-D-mannosidic linkages which are attached to an alpha-(1 leads to 6)-linked mannan back-bone chain. The alpha-(1 leads to 5)-linked D-arabinose residues located in the side chains were shown, by comparison of the immunochemical activities of the native and enzyme-degraded polysaccharides, to be the main immunodeterminants, as in the cell-wall arabinogalactan. There appeared to be variations in the ratio of arabinose and mannose residues, and also in the proportion of (1 leads to 2)-linked D-mannose units, depending on the individual strain; no (1 leads to 2)-mannosidic linkage was found in M. smegmatis arabinomannan. In addition to arabinomannan, a serologically inactive alpha-D-mannan ([alpha)D, +65 degrees approximately 68 degrees), whose structure may resemble that of the core mannan of the arabinomannan, was isolated as a copper hydroxide complex from the soluble fraction of disintegrated mycobacterial cells.


Purification and properties of human pancreatic deoxyribonuclease I.

Human pancreatic DNase I was purified extensively from duodenal juice of healthy subjects by a procedure including ammonium sulfate fractionation, ethanol fractionation, phosphocellulose fractionation, isoelectric focusing, and gel filtration. The final preparation was free of DNase II, pancreatic RNase, alkaline phosphatase, and protease. The enzyme had a molecular weight of approximately 30,000, as determined by gel filtration on Sephadex G-100, and showed maximum activity at pH 7.2-7.6. It required divalent cations for activity, and caused single-strand breaks by endonucleolytic attack on double- as well as single-stranded DNA molecules. The enzyme was inhibited by actin and bovine pancreatic DNase I antibody.


Carbonic anhydrase from spinach leaves. Isolation and some chemical properties.

Spinach carbonic anhydrase has been purified by modification and extension of a published method (Pocker, Y., and Ng. J. S. U. (1973) Biochemistry 12, 5127-5134), using (NH4)2SO4 precipitation and chromatography on DEAE-cellulose, agarose, and DEAE-Sephadex. The enzyme so obtained was homogeneous by criteria of both standard and sodium dodecyl sulfate polyacrylamide gel electrophoresis and of constant specific activity throughout the elution profile on DEAE-Sephadex chromatography. The enzyme has an apparent Mr of 212,000 by gel filtration on Sephadex G-200, a Mr of 26,000 by sodium dodecyl sulfate electrophoresis, and each of the subunits contains approximately 1 g atom of zinc. These data and the excellent correlation between the number of lysine and arginine residues per subunit, and the number of tryptic peptides obtained by peptide mapping, suggest that spinach carbonic anhydrase is an octamer consisting of identical or very similar subunits. Its amino acid composition is similar to parsley carbonic anhydrase; both contain large numbers of half-cystine residues relative to erythrocyte carbonic anhydrases. The spinach enzyme is devoid of disulfide bonds. The enzyme is stable around neutrality at -14 degrees, as a suspension in saturated (NH4)2SO4 solution.


Conformational adaptability of the active site of beta-galactosidase. Interaction of the enzyme with some substrate analogous effectors.

The action of different effectors, glycosides, and alcohols on the reactions catalyzed by beta-galactosidase is analyzed in this paper. Effectors as large as tri- and tetrasaccharides have no effect on the enzyme activity, suggesting that the binding site has rather small size. Most of the beta-galactosides produce a competitive inhibition. The other compounds assayed behave either as noncompetitive inhibitors, and they are deadened inhibitors, or as uncompetitive inhibitors which exhibit a better affinity for the chemical intermediate than for free enzyme; nearly all of them give transfer products. The analysis of the data indicates that the active center of beta-galactosidase is made up of two subsites: a galactose and a glucose subsite. This latter site is in a more favorable conformation in the galactosylenzyme than in free enzyme; possibly it might even by generated by the galactose binding. Conformational rearrangements of the active center deduced from the inhibition data have been directly observed by differential spectroscopy. The conformational adaptability of the enzyme and its consequence for the functional properties of beta-galactosidase are discussed.


Tubular elements - a new structure in blue-green algal cells.

New cell inclusions, 'tubular elements' are described from the blue-green alga Calothrix marchica. These structures are found in about 20% of all cells observed. They are 18-22 nm in diameter, apparently composed of sub-units of which the estimated number, as seen in transverse section, is 8-10.


Inhibition of platelet prostaglandin synthetase by oral aspirin.

Aspirin inhibits platelet function by permanently acetylating the cyclooxygenase that forms prostaglandins. We determined the sensitivity of platelets to aspirin in normal subjects by measuring [3H-acetyl]aspirin-susceptible cyclooxygenase in washed platelets obtained at various times after aspirin ingestion. A single 325-mg aspirin dose inactivated 89% of platelet cyclooxygenase. The inhibition persisted for 2 days suggesting that oral aspirin also inactivated megakaryocyte cyclooxygenase. Thereafter, active enzyme returned with a time-course reflecting platelet turnover (life-span 8.2+/-2 days). Single doses of 20-650 mg aspirin resulted in 34- greater than 95% inhibition after 24 h. Daily doses of 20-325 mg aspirin for brief periods produced 61- greater than 95% inactivation when measured 24 h after cessation of the drug. Platelet cyclooxygenase is more sensitive to inactivation by aspirin than enzyme in sheep seminal vesicles.


The physiological role of thyrotropin-releasing hormone in the regulation of thyroid-stimulating hormone and prolactin secretion in the rat.

The physiological role of thyrotropin-releasing hormone (TRH) in the regulation of thyrotropin (thyroid-stimulating hormone, TSH) and prolactin (Prl) secretion has been assumed but not proven. Stimulation of their release requires pharmacologic doses of TRH. Lesions of the hypothalamus usually induce an inhibition of TSH secretion and an increase in Prl. To determine whether TRH is essential for TSH and Prl secretion in the rat, 0.1 ml of TRH antiserum (TRH-Ab) or normal rabbit serum was administered to normal, thyroidectomized, cold-exposed, and proestrus rats through indwelling atrial catheter. Serum samples were obtained before and at frequent intervals thereafter. Serum TSH concentrations in normal, thyroidectomized, cold-exposed, and proestrus rats were not depressed in specimens obtained up to 24 h after injection of normal rabbit serum. In contrast, serum TSH was significantly decreased after the administration of TRH-Ab in all normal (basal, 41+/-8 muU/ml [mean+/-SE]; 30 min, 6+/-2; 45 min, 8+/-3; 75 min, 4+/-2); thyroidectomized (basal, 642+/-32 muU/ml; 30 min, 418+/-32; 60 min, 426+/-36; 120 min, 516+/-146); coldstressed (basal, 68+/-19 muU/ml; 30 min, 4+/-3; 180 min, 16+/-8); and proestrus (basal, 11 a.m., 57+/-10 muU/ml; 1 p.m., 20+/-3; 3 p.m., 13+/-4; 5 p.m., 19+/-3) rats. However, 0.1 ml of TRH-Ab had no effect on basal Prl concentrations in normal or thyroidectomized rats and did not prevent the Prl rise in rats exposed to cold (basal, 68+/-7 ng/ml; 15 min, 387+/-121; 30 min, 212+/-132; 60 min, 154+/-114), or the Prl surge observed on the afternoon of proestrus (basal 11 a.m., 23+/-2 ng/ml; 1 p.m., 189+/-55; 3 p.m., 1,490+/-260; 5 p.m., 1,570+/-286). These studies demonstrate that TRH is required for TSH secretion in the normal, cold-exposed and proestrus rat and contributes, at least in part, to TSH secretion in the hypothyroid rat, but is not required for Prl secretion in these states.


Leprosy and the serodiagnostic test for tuberculosis.

Sera from patients with leprosy agglutinated killed H37Rv Mycobacterium tuberculosis to varying degrees. Higher titres were found in association with active disease but there was no difference between patients at the lepromatous or at the tuberculoid ends of the clinical scale. A sharp rise in titre occurred during the active phase of an episode of erythema nodosum leprosum.


Cross infection in a surgical ward caused by Pseudomonas aeruginosa with transferable resistance to gentamicin and tobramycin.

An outbreak of gentamicin- and tobramycin-resistant Pseudomonas aeruginosa infection occurred in a surgical ward over a three-month period. Resistant Ps. aeruginosa strains with the same serological, phage, and pyocin type were cultured from the urine of six patients. Identical organisms were found on urine bottles, bedpans, and the hands of attendant staff. Inadequate disinfection played a major role in cross-infection. Isolates of the epidemic strain from each of the patients and of an unrelated but similarly resistant Ps. aeruginosa from one of them could transfer resistance to a recipient strain of Ps. aeruginosa. Resistance to gentamicin, kanamycin, tobramycin, sulphonamides, and mercuric chloride was determined by R factors belonging to Pseudomonas incompatibility group P-3. Aminoglycoside resistance was due to acetylation.


Aminoglycoside cross-resistance patterns of gentamicin-resistant bacteria.

Ten strains each of Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa were habituated to gentamicin by serial passage in antibiotic containing medium. Complete cross-resistance to streptomycin, neomycin, kanamycin, and tobramycin in a linear proportional fashion was demonstrated at all stages of habituation. Most strains of Staph. aureus and Ps. aeruginosa showed a greater increase in resistance to gentamicin than to the other three aminoglycosides. E. coli required more transfers to reach the same degree of resistance than did the other two species. Reversion to greater susceptibility to gentamicin took place after serial passage on antibiotic-free media. 'Wild' gentamicin-resistant strains showed no such proportionality of resistance to kanamycin, neomycin, or streptomycin. But many of these strains showed a proportional increase in resistance to tobramycin.


The retinotopic organization of area 17 (striate cortex) in the cat.

The location and retinotopic organization of visual areas in the cat cortex were determined by systematically mapping visual cortex in over 100 cats. The positions of the receptive fields of single neurons or small clusters of neurons were related to the locations of the corresponding recording sites in the cortex to determine the representations of the visual field in these cortical areas. In this report, the first of a series, we describe the organization of area 17. A single representation of the cat's entire visual field corresponds closely to the cytoarchitectonically defined area 17. This area has the largest cortical surface area (380 mm2) and the highest cortical magnification factor (3.6 mm2/degree2 at area centralis) of all the cortical areas we have studied. There was perfect agreement between the borders of area 17 determined electrophysiologically and cytoarchitecturally. This area contains a first order transformation of the visual hemifield in which every adjacent point in the visual field is represented as an adjacent point in the cortex. Some variability exists among cats in the extent and retinotopic representation of the visual field in area 17.


Effects of prestriate, inferotemporal, and superior temporal sulcus lesions on attention and gaze shifts in rhesus monkeys.

The role of extrastriate cortical areas in selective attention was studied in 12 rhesus monkeys. Animals learned a series of color-form pattern discrimination problems, with either color or form cues relevant. After each problem was mastered, correct behavior required a shift in attention, i.e., that responses be made to be previously irrelevant dimension. On some problems shifting attention required that the animal maintain the same fixation; on other problems the color and form cues were separated in space, and the attention shift presumably required a shift in gaze. Matched groups of animals with inferotemporal, prestriate, or superior temporal sulcus lesions, and normal controls, differed significantly in their ability to shift attention. Analyses of inferred stages in attention shift showed that different processes  were disturbed in the three lesion groups. Results are discussed in terms of cortical substrates for "looking" and "seeing".


Antibacterial effect of the lactoperoxidase system on psychotrophic bacteria in milk.

Activation of the antibacterial lactoperoxidase system in milk, i.e. increasing the thiocyanate concentration to 0.25 mM and adding an equimolar amount of H2O2, results in a substantial reduction of the bacterial flora and prevents the multiplication of psychrotrophic bacteria for up to 5 d. This treatment has no effect on the physico-chemical properties of milk and does not lead to the accumulation of resistant bacteria. The practical application of the lactoperoxidase system in prolonging the storage period of raw milk at low temperatures is discussed.


The acute effects of tourniquet ischemia on tissue and blood gas tensions in the primate limb.

Tourniquet ischemia results in tissue hypoxia which has been measured indirectly by blood gas analysis. The Medspect mass spectrometer allows direct measurement of gas tension in different tissues and may provide more useful information regarding safe tourniquet times. Calibrated Teflon catheters were inserted into the subcutaneous tissue (11 animals), tibial medullary cavities (bone) (nine animals), and tibialis anterior muscles (10 animals) in both lower extremities of anesthetized stumptail monkeys. Tourniquet ischemia was maintained for 1 hour at 400 mm Hg. Tissue and venous blood gas tensions were recorded from both limbs for 1 1/2 hours. Comparisons between gas tensions in each tissue group were made on the basis of their percentage change from control values. In the ischemic limb within 5 minutes muscle PO2 fell 42 +/- 4% (p is less than 0.001), whereas bone and subcutaneous PO2 dropped 25 +/- 4% (p is less than 0.05). Blood PO2 fell 29 +/- 3% and differed only from that of muscle (p is less than 0.01). One hour after tourniquet inflation, blood PO2 levels has fallen 90 +/- 5% (p is less than 0.001) from their control. Changes in tissue PCO2 were less dramatic and did not vary significantly from those recorded in venous blood. After deflation, blood PO2 exceeded its control by 20 +/- 6% (p is less than 0.005) in 5 minutes, but tissue tensions remained 50 +/- 6% below their control values. These studies indicate that tissue gas tensions are a more sensitive indicator of tourniquet hypoxia than are blood gases within the ischemic extremity.


Common host antigens in laboratory rats infected with the metacestodes of Taenia crassiceps.

Laboratory rats were infected by intra-peritoneal injection with the metacestodes of Taenia crassiceps. A soluble extract was prepared from the metacestodes removed from the peritoneal cavity of the rats. Double-diffusion, immuno-electrophoresis and fluorescent labelled antibody staining techniques were used. The extract tested against rabbit anti-normal rat serum was found to contain an antigen common to both the host and the parasite.


Fixation variables in horseradish peroxidase neurohistochemistry. I. The effect of fixation time and perfusion procedures upon enzyme activity.

In a series of neurohistochemical experiments the effect of aldehyde fixation upon the detection of horseradish peroxidase (HRP) was examined. These experiments demonstrated that: a) Increments in fixation of as little as 1 hr significantly decreased the number of labeled neurons; 12-hr fixation abolished HRP activity in many neuronal populations and significantly reduced the apparent size of the injection site. b) This negative fixation effect was greatest where the HRP concentration was low (e.g. in small, lightly labeled neurons) but was still evident in areas of high concentration (e.g. large, heavily labeled neurons). c) This effect was also most prominent when a less sensitive diaminobenzidine histochemical procedure was employed but was still apparent with a more sensitive benzidine dihydrochloride procedure. d) Immersion of the brain in fixative after perfusion produced a greater attenuation of HRP activity in more superficial areas. e) Immersion of the brain in buffer to terminate fixation produced a prolonged and unpredictable gradient of fixation. f) Excess, unbound fixative inhibited the histochemical reaction per se and had to be removed from the tissue but prolonged washing did not resurrect enzyme activity which was lost by fixation. To obviate these problems and optimize HRP enzyme activity a new perfusion-fixation procedure was developed. It entails 30 min fixation by perfusion which is terminated by a subsequent 30 min perfusion with cold sucrose-fuller to wash out unbound fixative. This allows the tissue to be processed immediately, produces a uniform and morphologically adequate fixation, and minimizes the negative effects of fixation on HRP enzyme activity.


Observations on the effects of formaldehyde on cockroaches and their flora: II. Prolonged survival of cockroaches drinking formaldehyde or glutaraldehyde solutions.

Adult cockroaches were found to survive up to 22 weeks when provided with 1% Formalin (0.4% formaldehyde) in lieu of drinking water. Given 4% Formalin or 2% glutaraldehyde they survived up to 41 days. During the experiments eggs were laid and hatched and the offspring continued to grow. Combined with surface disinfection, this may hold out a simple method of rendering adult cockroaches gnotobiotic or even axenic.


Binding studies on heavy-light chain recombinant hybrid immunoglobulins A derived from murine myeloma anti-fructofuranans.

Heterologous H-L chain recombinants derived from homogeneous murine myeloma immunoglobulins with anti-inulin specificity were found to bind inulin-related oligosaccharides with affinities closely paralleling those of the L chain donors.


Stimulation of an enhanced in vitro immune response by a synthetic adjuvant, muramyl dipeptide.

Various subcellular bacterial fractions are known to enhance immune responses and serve as potent adjuvants. Muramyl dipeptide (MDP), a synthetic adjuvant mimicking a component of mycobacterial cell walls, enhances humoral immunity to soluble antigens and can increase macrophage cytotoxicity toward mastocytoma cells in vitro. In the present study MDP was found to enhance the hemolytic antibody plaque response of normal mouse spleen cells in vitro to SRBC at a level equal to or greater than that induced by Escherichia coli lipopolysaccharide. Furthermore, MDP was found to enhance the antibody response to SRBC nonspecifically in unimmunized spleen cell cultures, suggesting that similar to LPS the synthetic dipeptide may induce a generalized clonal expansion of committed lymphocytes and thus serve as a "polyclonal activator." MDP also enhanced the immune responsiveness of normal splenocytes to suboptimum concentrations of SRBC, indicating that this material may be useful in enhancing immunity in situations where there would normally be a poor immune response.


Variation of the recombination fraction in Drosophila melanogaster females.

The distribution of the recombination fraction between two distal, autosomal loci, plexus and orange, was investigated in reciprocal crosses between the marker stock and each of two wild-type strains, Harwich and Ottawa. The dysgenic cross between Harwich males and marker females exhibited high sterility, low fecundity, and a non-normal distribution of the recombination fraction. Neither the distribution of the reciprocal Harwich cross nor those of the two Ottawa crosses deviated from normal. In general, the results support the view that the recombination fraction is a variable.


Differential immunosuppressive effects of ascites fluid from mastocytoma-bearing mice on primary vs secondary immunocyte responses.

The effects of immunosuppressive ascites fluids from mastocytoma-bearing mice on the primary vs secondary immune response to sheep red blood cells (SRBC) was examined. Injection of mice with ascites fluid from tumor-bearing mice markedly depressed the primary immune response of normal syngeneic mice challenged with SRBC. However, there was a preferential depression of the 19S IgM antibody response as compared with the 7S IgG response. Injection of ascites fluid shortly before secondary immunization of mice with SRBC also resulted in depressed IgM PFC responses but only a slight to moderate depression of IgG PFC. Treatment of mice with the ascites fluid before primary immunization had little if any effect on the secondary IgG PFC response, although the IgM response was moderately depressed. These results indicate that the immunosuppressive factor(s) present in the ascites fluid of mastocytoma-bearing mice has a differential effect on distinct classes of immunocytes. Those immunocytes or their precursors involved in formation of low efficiency 7S IgG antibody are more resistant to immunodepression. Such differences appear due to different sensitivities of cells involved in the immune response system.


Uranyl-labelled antibody (ULA) method and its application for Rh antibodies.

The existent labelling materials for studies of antigen--antibody interaction at ultrastructural level, namely ferritin and peroxidase, because of their large molecular size do not fulfill all requirements of excellent markers for electron microscopy (EM). Uranyl acetate has a molecule 354 times smaller than IgG and its uranium atom is electron-dense. These physical characteristics of uranyl acetate make it a labelling material par excellence as described in this article. Quantitative and qualitative studies of Rh antigen-antibody interactions are for the first time presented at the ultrastructural level, and the application of the uranyl-labelled antibody (ULA) method for weak antisera (dilutions 100 to 1000 time higher than the Coombs range of sensitivity) is demonstrated. The ULA method opens a new era for studies of antigens, antibodies and their interactions because it will demonstrate visibly details of the antigen-antibody interaction and is especially suitable for studies of weak antisera.


The toxicity of melanin precursors.

The quinone intermediates resulting from tyrosinase-mediated oxidation of tyrosine were evaluated as sulfhydryl reagent inhibitors of purified calf thymus DNA polymerase alpha in order to determine which of these might be cytotoxic. Dopachrome and an oxidation product of 2,4,5-trihydroxyphenylalanine were relatively ineffective as inhibitors of DNA polymerase alpha. On the other hand, a dopaquinone analogue, 4-(2-N-acetylaminoethyl)-1,2-benzoquinone, synthesized from N-acetyl dopamine, was demonstrated to have marked affinity for this sulfhydryl enzyme. This property was shared by 1,2-benzoquinone. These studies point to dopaquinone as a significant toxic metabolite in melanin biosynthesis.


Detection of immune complex-like materials in cancer patients' sera: a comparative study of results obtained with the C1q deviation and C1q binding tests.

In a collaborative study involving three laboratories, randomly coded sera from 47 patients and healthy donors were tested for soluble immune complexes by two versions of the C1q BT and by the C1q DT. Analysis of ranked data showed a close correlation between results obtained in all laboratories as well as good reproducibility on testing coded duplicate samples included in each panel of sera. However, with the use of values for normal donors established independently in each laboratory, the tests did not always agree in discriminating normal from abnormal sera, particularly with sera from cancer patients. Results reflected to some extent the methods used to inactivate the endogenous C1 complex in the test sera. Studies of 130 additional cancer patients revealed that 44 (34%) gave abnormal C1q BT values when inactivated with EDTA but only 35 (27%) were abnormal when heat inactivated (56 degrees for 30 min). Similarly 12 of 65 sera (18%) from patients with nonneoplastic diseases and 10 of 80 (13%) from healthy donors gave significantly abnormal results after addition of EDTA. Eight (12%) of the disease and six (8%) of the healthy controls' sera were similarly outside normal limits when heat inactivated. Repeated freezing and thawing of sera or changes in the concentration of PEG used to precipitate complex-bound 125I-C1q influenced C1q BT results more than duration of storage at -70 degrees C or changes ascribed to presence or absence of rheumatoid factors and CRP.


Porcine Willebrand factor: a population of multimers.

Purified porcine Willebrand factor was analyzed by agarose-sodium DodSO4 electrophoresis. Multiple forms of the protein were found in a series of increasing molecular weights. A molecular mass calibration curve was constructed with fibrinogen (3.4 X 10(5) daltons), IgM (1 X 10(6) daltons), and glutaraldehyde-crosslinked IgM polymers (2, 3, and 4 X 10(6) daltons). As measured by this procedure, the apparent molecular weight of Willebrand factor polymers ranged from 1.1 X 10(6) to 2.1 X 10(7). Each member of the series differed from one another by approximately 1.5 to 1.9 X 10(6) daltons, indicating that members of the series were polymers of 6-mers to 8-mers of the 2.3 X 10(5) dalton subunit. Various purification procedures, used to isolate Willebrand factor active in inducing platelet aggregation, were seen to fractionate the polymers, in part, on the basis of size. The same purification procedures, when applied to procine von Willebrand plasma, failed to yield protein of molecular weight greater than 1.1 X 10(6).


Metabolism of 5-methyltetrahydrofolate by Lactobacillus casei.

The metabolism of 5-[Me-14 C]methyltetrahydrofolate in Lactobacillus casei proceeded oxidatively with incorporation of label into purine and thymidylate derivatives. No labelled methionine was formed. (l)-5-Methyltetrahydrofolate, the natural isomer, was not a substrate for the L. casei folylpoly-gamma-glutamate synthetase although the unnatural (d)-isomer was slowly metabolized to the diglutamate form.


Rate-limiting steps in folate metabolism by Lactobacillus casei.

Oxidation of 5-methyltetrahydrofolate to 5,10-methylenetetrahydrofolate was the rate-limiting step in 5-methyltetrahydrofolate metabolism by Lactobacillus casei. The limiting steps in the utilization of suboptimal levels of folate by L. casei were related to the ability of folates to function in purine and/or thymidylate biosynthesis. Folates with glutamate chains of up to at least seven residues were substrates for these biosynthetic enzymes, and comparisons of bacterial growth yields with transport rates for these folates indicated that the polyglutamates were more effective substrates in purine and thymidylate synthesis than the corresponding pteroylmonoglutamates. Lactobacillus casei contained low levels of a B12-independent, pteroylpolyglutamate-specific methionine synthetase. Its methylenetetrahydrofolate reductase also functioned more effectively with pteroylpolyglutamate substrates.


Purification and properties of a protease with elastase activity from Pseudomonas aeruginosa.

The isoelectric points of three proteases (I, II and III), separated from culture supernatants of Pseudomonas aeruginosa strain PAKS-I by isoelectric focusing, were 8.5, 6.6 and 4.5 respectively. Collagenase activity was not detected. More than 75% of the extracellular protease activity of this strain was due to protease II. This enzyme also possessed elastase activity. When purified by ammonium sulphate precipitation, isoelectric focusing and gel chromatography, protease II showed one band on disc electrophoresis and one band on conventional immunoelectrophoresis. The pH optimum, stability and effect of inhibitors and substrate concentration were examined. The molecular weight was 23000 +/- 5000. Protease II was lethal for mice when injected intraperitoneally at a high dose (minimum lethal dose 0.1 mg). Dermonecrosis and subcutaneous haemorrhages were produced in new-born mice upon subcutaneous injection of 10 microgram protease II. A sensitive test for cytotoxicity showed no evidence of cytoplasmic membrane damage to HeLa cells or human diploid embryonic lung fibroblasts by protease II. Morphological changes similar to those produced by trypsin were found.


Protease-deficient mutants of Pseudomonas aeruginosa: pleiotropic changes in activity of other extracellular enzymes.

Mutants of Pseudomonas aeruginosa strain PAKS-I which are defective in the formation of extracellular protease activity have been characterized. The mutants produced between approximately 1 and 25% of the protease activity of the wild type and no strains completely lacking extracellular protease were found, even after repeated mutagen treatment. Most mutants also had changed activities of extracellular staphylolytic enzyme, lipase and lecithinase. Four of 13 mutants were unable to release alkaline phosphatase and staphylolytic enzyme into the medium in contrast to the wild type. Serotype, phage type and biochemical reactions were essentially unchanged. The results indicate that some of the mutations affected the cell envelope structure of function leading to decreased ability to release extracellular proteins, and that other mutations possibly affected a common regulatory mechanism for extracellular enzymes.


Biosynthesis of thiamin in Bacillus subtilis. Isolation of mutants accumulating 4-amino-5-hydroxymethyl-2-methylpyrimidine phosphate.

Thiamin-deficient mutants of Bacillus subtilis were characterized by their growth responses to the pyrimidine and thiazole moieties of the vitamin molecule and by cross-feeding tests. All mutants growing on the thiazole moiety and all mutants with an absolute requirement for thiamin fed all those growing on the pyrimidine moiety. No other cross-feeding effects were observed. From the culture fluid of a mutant growing on the thiazole moiety, two compounds were isolated which supported growth of mutants requiring the pyrimidine moiety. These compounds were identified by chromatographic, bioautographic and spectrophotometric procedures as 4-amino-5-hydroxymethyl-2-methylpyrimidine and its monophosphate derivative.


Vascular reactivity in the primate brain after acute cryogenic injury.

The effects of an acute cryogenic injury on cerebral flow (CBF) and cerebral vascular reactivity were studied in 12 anaesthetised, ventilated baboons. Autoregulation, defined in this study as intact with a greater than 20% change in cerebrovascular resistance in response to a change in cerebral perfusion pressure, was tested before the lesion by arterial hypotension. Intact autoregulation was found in half the animals, but all animals showed an increase in CBF with hypercarbia. The cryogenic lesion was followed by a marked rise in intracranial pressure, and a fall in CBF which was only partly related to the status of autoregulation beforehand. After injury, arterial hypertension caused an increase in cerebrovascular resistance of more than 20% in half the animals. This response was not related to the presence of autoregulation before the lesion, and was accompanied by a greater impairment of the cerebrovascular response to carbon dioxide, more severe brain oedema, and lower cerebral oxygen consumption, than in the remaining baboons which had a pressure passive response to arterial hypertension. This study confirms that the failure of CBF to increase with arterial hypertension may indicate severe brain damage rather than intact physiological autoregulation.


Systemic vascular responses to increased intracranial pressure. 2. The 'Cushing' response in the presence of intracranial space-occupying lesions: systemic and cerebral haemodynamic studies in the dog and the baboon.

Continued expansion of an artificial space-occupying lesion produced further increases in mean supratentorial and infratentorial pressures associated with increases in mean arterial pressure, heart rate, and systemic vascular resistance-the `Cushing' or systemic hypertensive response. These primary changes resulted in an increase in transtentorial pressure gradient and a decrease in arrhythmia index. Immediately before the onset of the systemic hypertensive response, supratentorial perfusion pressure was low, and the period of systemic hypertension did not appear to produce any worthwhile improvement in the perfusion pressure or in the blood flow in the supratentorial compartment. The studies demonstrated also that the systemic hypertensive response was a pre-terminal event and was followed rapidly by circulatory failure.


An iodinated catecholamine congener for brain imaging and metabolic studies.

The iodinated O-methylated catecholamine congener, 4-iodo-2,5-dimethoxyphenylisopropylamine (4-I-DPIA), has potential as a new agent for imaging and metabolic studies of the brain and lung. The organ distribution and brain uptake of radioiodine-labeled 4-I-DPIA were studied in the dog and monkey by whole-body scanning, gamma-camera scintigraphy, and organ assay. The brain takes up 2% of the injected dose, with a half-time of 8 sec in the monkey, and the lung takes up 11.8%. An unusual finding was a concentration in the retina, five times that in any other CNS tissue. 4-I-DPIA may have potential in the imaging of normal brain tissues and thereby delineating nonfunctional areas damaged by infarction, trauma, or malignancy, and may also be useful in metabolic studies of catecholamine function. Adequate radioactivity can theoretically be administered with a quantity of 4-I-DPIA 1/10,000 of the pharmacologically active levels. The agent may also find application in lung imaging because of the high pulmonary uptake.


[32P] diphosphonate dose determination in patients with bone metastases from prostatic carcinoma.

In an initial safety study, phosphorus-32 (as diphosphonate) was administered intravenously to five patients with painful bone metastases from prostatic carcinoma; two patients received 9 mCi and three were given 3 mCi. Hematological, biochemical, ECG, x-ray, bone-scan data, and clinical observation, were followed for 2 mo. At both dose levels, bone-marrow depression was noted. One of the patients, who received 9 mCi, had only a slight dip in the levels of circulating white blood cells and platelets. The other 9-mCi patient was the only one with discrete metastases by bone scan; he had bone-marrow depression, from which he recovered, and was the only one of the five who had relief of bone pain.


Schedule of protein ingestion and circadian variations of glycogen phosphorylase, glycogen synthetase and phosphoenolpyruvate carboxykinase in rat liver.

The circadian rhythms of liver glycogen and hepatic activity of glycogen synthetase (GS), glycogen phosphorylase (GP) and phosphoenolpyruvate carboxykinase (PEPCK) were studied in adult male rats. The rats either received a mixed diet ad libitum (10% protein) or a protein meal (1.85 g protein) given at 09:00 or 21:00 hours, with free access to a protein-free diet (separately-fed). When the protein meal was ingested at 09:00 hours it was followed by a drop in liver glycogen and a persistent daylight increase in GP and PEPCK activities, this phenomenon being attenuated when proteins were ingested during darkness (21:00 hours). Moreover in the latter case, the circadian rhythm of liver glycogen was modified (glycogen accumulation occurring later) and the protein meal ingestion was followed after a transient decrease by a high and sustained GS activity during a long period (12 hours). The drop in the hepatic glycogen level and the unusually long daylight period of sustained GP and PEPCK activities in separately-fed rats consuming the protein meal at 09:00 hours suggests that, in this case, part of the ingested nitrogen could have been catabolized and used for gluconeogenesis, thus explaining our previous observation of lower nitrogen retention observed in this group of rats.


Respiratory manifestations and skin reactivity in the detergent industry.

Workers at two continental European enzyme detergent plants were examined to determine respiratory symptomatology, pulmonary function abnormalities and skin reactivity to four enzymes through use of a standardized questionnaire, flow-volume curves and provocative acetylcholine, single breath carbon monoxide, and prick tests. In both plants, the frequency of bronchitis was similar but the frequency of asthmatic or rhinopharyngitic symptoms was different. Symptomatology was more frequent in female detergent workers than among the clerical staff but no difference among men in different jobs was noted. Very few spirometric abnormalities were observed but bronchial hyperactivity was more frequent in packaging and warehouse workers. No correlation between DLco/AV and of work or skin reactivity was found. Skin reactivity (about 10% in both plants) and duration of employment were not related to symptomatology nor functional abnormalities.


Quantitation of blood flow after anterior maxillary osteotomy: investigation of three surgical approaches.

The radioactive microsphere method was used to quantitate preoperative and postoperative blood flows in macaque monkeys when three different surgical approaches for anterior maxillary osteotomy were performed. Despite distinct variations in flap designs among the experimental groups, preoperative and postoperative determinations of blood flow were essentially unchanged. Results of the study suggest that a palatal, labial, or combined mucoperiosteal pedicle should be adequate to preserve the flow of blood to tissues in the osteotomized segment.


Mechanisms involved in mandibular condylopathy secondary to intraarticular injections of glucocorticoids.

Pharmacologic doses of triamcinolone hexacetonide were injected intramuscularly or intraarticularly in immature Papio papio baboons. The mandibular condyle served as a model for histologic examinations concerning the effect of glucocorticoid hormone on cartilage and bone. Biochemical examinations of blood and urine indicated the development of distinct hypophosphatemia, hypercalcemia followed by hyperphosphaturia, and hypocalciuria. Serum alkaline phosphatase activity rose during the initial phases of the experiment but decreased considerably after the sixth injection of the hormone. Hyperglycemia and an increase in serum amylase were noticed along with signs of moderate metabolic acidosis. Histologic examinations disclosed signs of severe destruction of cartilage and bone. By the sixth intraarticular injection, definite fibrillation was noted in the articular cartilage, followed by complete disappearance of cartilage. The subchondral bone appeared to be adversely affected by the hormone as it lost its typical lamellar organization and attained the characteristics of woven bone. The condyle showed clear signs of fibro-osseous transformation, with fibrosis as the dominant structural feature. The preceding biochemical and morphologic findings are indicative of parathyroid hyperactivity.


Disaccharidase activities in small intestinal mucosa in patients with cystic fibrosis.

The disaccharidase activities in small intestinal biopsies were related to the morphology of the mucosa and the ages of 63 patients with cystic fibrosis and 177 healthy control subjects of Caucasian origin. In patients with CF and in the healthy control subjects under 5 years of age with normal intestinal mucosa, no low lactase activity was found. In those patients with CF who were over 5 years of age, one group had high and one group had low lactase activity, as occurs in healthy Caucasian control subjects of the same age. This finding supports the view that in patients with CF, lactase deficiency is not related to the disease entity. In patients with or without CF who had the same degree of mucosal atrophy, the decrease of disaccharidase activities followed the same pattern, indicating that enzyme activities are affected to the same extent by the damage of the mucosa. In patients with CF with pancreatic insufficiency, the disaccharidase activities were significantly (P less than 0.001) higher when compared to those in control subjects of the same age and ethnic group, although the increase was not uniform in all patients with cystic fibrosis.


Quantitative assessment of free gingival grafts with and without periosteum and osseous perforation.

Forty free gingival grafts were performed on 5 adult Rhesus monkeys as follows: 10 grafts were placed on recipient sites with periosteum; 10 grafts on periosteum and perforated cortical plate; 10 grafts on bared bone; and 10 grafts on perforated bared bone. All free autografts (7 X 5 X 1 mm) were taken from the palatal mucosa and were sutured in place. Clinical evaluation and photographs were taken at 1, 2, 4, 8, 12, 16, 20 and 24 weeks postoperatively. At 24 weeks, a device was used to measure quantitatively the amount of force necessary to pull the graft 2 mm away from the underlying bone. Biopsy of two grafts of each type were taken for histologic examination. The following conclusions were drawn: 1. There was no difference in the survival or healing pattern of grafts placed on periosteum and grafts placed on bared bone.


GLC determination of the polyvalent saluretic uricosuric agent (2-cyclopentyl-6,7-dichloro-2-methyl-1-oxo-5-indanyloxy)acetic acid in biological fluids.

A specific and quantitative GLC method was developed for the determination of (2-cyclopentyl-6,7-dichloro-2-methyl-1-oxo-5-indanyloxy) acetic acid, a novel saluretic uricosuric agent, in biological fluids. The procedure involves the addition of an internal standard to the biological specimens followed by extraction of the acids into benzene at pH 1. The extracted acids, following back-extraction into base and reextraction into methylene chloride at an acidic pH, are converted to the respective methyl esters by reaction with ethereal diazomethane. The sensitivity of the method is such that 2 microgram of material can be detected per aliquot of plasma or urine. In the 2.5-50-microgram/ml range, recoveries were 98.1 +/- 9.6% (plasma, n = 157) and 99.3 +/- 6.4% (urine, n = 181). GLC-mass spectrometric techniques established analysis specificity.


Continuous intravenous naltrexone effects on morphine self-administration in rhesus monkeys.

Rhesus monkeys, surgically prepared with intravenous catheters, were given opportunities to self-administer morphine for 3 days, methamphetamine for 2 days and saline for 2 days in a constantly repeating cycle. Access to drugs was limited to a 15-minute period every 4 hours. After stable base-line self-administration rates, saline or various concentrations of naltrexone were infused continuously through the catheter. In the first phase of the study each concentration of naltrexone was infused for 4 weeks (separated by 3 weeks of saline) while the dose of morphine available for self-administration was held constant at 8 microgram/kg/injection. Stable naltrexone dose-related suppression of morphine self-administration occurred throughout each 4-week infusion. In the second phase of the study, various doses of morphine were made available for self-administration during 6- to 8-week continuous infusions of saline or various concentrations of naltrexone. The dose-effect curve relating self-administration rate to morphine dose per injection shifted to the right and decreased in maximum as the rate of infusion of naltrexone increased. Methamphetamine and saline self-administration rates were unaffected by naltrexone.


Relationship between intrapulmonary airway diameter and smooth muscle tone in excised lungs.

1. Intrapulmonary bronchi in excised dog lungs were outlined with tantalum dust and stereoscopic radiographs taken during deflation and inflation of the lung after rinsing with solutions of saline, histamine, isoprenaline or EDTA. Dimensions of airways were calculated from measurements of the stereoscopic X-ray images. 2. After treatment with EDTA to minimize bronchial smooth muscle activity, airway diameters increased at all transpulmonary pressures (Ptp) and lung volumes relative to their diameter after treatment with histamine; airway hysteresis in relation to Ptp decreased. 3. At low lung volumes, the per cent increase from histamine to EDTA for airways of different sizes was the same (24-30%) but at high volumes (30 cm H2O distending pressure) the dilatation induced by EDTA was 30% for airways less than 3.0 mm diameter and 13% for those greater than 5.0 mm diameter. 4. Even at high lung volumes, intrapulmonary airways are free to constrict or dilate in spite of the stiffness of the supporting parenchyma.


Dual capacity for nutrient uptake in Tetrahymena. II. Role of the two systems in vitamin uptake.

Previously, we found that a mutant strain of Tetrahymena pyriformis without food vacuoles failed to grow unless the nutrient media were richly supplemented with vitamins and trace metals. Here we show that calcium folinate alone can replace the extra vitamin supplementation. The mutant requires approximately 90-fold higher concentration of folinate than the wild-type cells to give similar growth responses in a chemically defined medium. We infer that the food vacuole is an important route of uptake for this vitamin in the wild-type cells. We found no difference between mutant and wild-type cells in their requirements for nicotinic acid, pantothenic acid, riboflavin-monophosphate, and pyridoxal. We infer that an extravacuolar route contributes importantly to uptake of these 4 compounds.


Isolation and properties of flagella of trypanosomatids.

A procedufe is described for the isolation of flagella of Crithidia fasciculata, Herpetomonas samuelpessoai and Leishmania tarentolae in a highly purified state and giving reasonably good yield. The 3 types of flagella give a similar electrophoretic pattern of proteins. It is shown that H. samuelpessoai and, to a lesser extent, C. fasciculata flagella confer protection against Trypanosoma cruzi infection.


Exacerbation of experimental Trypanosoma cruzi infection in mice by concomitant malaria.

The effect of malaria on the chronic phase of Chagas' disease was investigated in mice. The animals were given Plasmodium bergheri-infected red blood cells 2 to 12 months after their initial inoculation with trypomastigotes of 3 different strains of Trypanosoma cruzi (Y. CL and Gilmar). in all the experiments carried out with one of the strains (CL), a somewhat variable but always considerable percentage of mice (average 39%) relapsed in to the acute phase of Chagas' disease. This relapse was characterized by a significant increase in the number of circulating trypomastigotes. Recrudescence was observed also with a 2nd strain of T. cruzi (Gilmar), which is similar in many aspects to the CL strain, e.g. the morphology of blood stages, curved of parasitemia and susceptibility to antibodies in vitro. In mice whose chronic phase was induced by trypomastigotes of the Y strain, malaria infections did not induce a typical acute phas with high parasitemia by T. cruzi. Bloodstream forms of Y parasites differ from those of CL and Gilmar strains morphologically as well as immunologically, i.e. only the Y strain is easily agglutinated and partly inactivated by specific immune serum. In light of this and other known characteristics of the strains used in the present work, the author speculates on mechanisms which allow malaria infections selectively to suppress acquired host resistance to certain strains of T. cruzi.


Euglena: the photoreceptor system for phototaxis.

The photoreceptor structures (eyespot-paraflagellar body-flagellum) for Euglena phototaxis were investigated by electron microscopy. The paraflagellar body--the photoreceptor--is a highly ordered crystalline lamellar structure. Optical diffraction of the electron micrographs and resulting filtered images of the paraflagellar body suggest that it is formed of rods in a helical arrangement. The action spectra for phototaxis, the in situ spectrum by microspectrophotometry of the paraflagellar body, and flavin analysis of the organism indicate that the photoreceptor molecule is a flavoprotein. The phototaxis action spectrum is similar to the spectrum for O2 evolution and implies that similar molecules participate in the photo processes. As a result, a photochemical scheme is suggested in which a photo-excited flavin and a cytochrome participate in the photoprocess. The photochemistry and photoreceptor structures for Euglena phototaxis are likened to a photoneuro sensory cell.


Antigens of the subcellular fractions of Trypanosoma cruzi. II. Flagellar and membrane fraction.

A method for isolation of membrane and flagellar fractions from Trypanosoma cruzi epimastigotes, and descriptions of their ultrastructural characteristics and antigenic activity are presented. Electron-microscopic observation revealed that the membrane fraction resembled vesicles, as occurs frequently when the pressure-depressure method is used for cell disruption. The flagellar fraction revealed flagella and some membranes, both related and unrelated to the flagellar structure. Flagellar and membrane fractions had 5 and 3 precipitin lines respectively against anti-whole homogenate of T. cruzi, in double diffusion tests. One of the precipitin bands common to both fractions gave a reaction of identiy with one precipitin line of the microsomal fractions, that was not present in the mitochondrial, nuclear and cell sap fractions. Data from tests of protective activity activity obtained during a period of 3 years with whole homogenate and F and M fraction are also presented Protective activity against lethal challenge doses of trypomastigotes is strongly associated with the flagellar fraction.


[Ureteral diverticulosis (author's transl)].

We report here two new cases of ureteral diverticulosis. This condition is found usually in the man of the fifties who has most frequently an urinary tract infection or a prostatic adenoma. The roentgen findings are multiple out-poutchings of the lumbar ureter. Several factors are combined (constitutionnal, urinary tract infection, disordered ureteral-motility) to explain the creation of diverticules in the areas of weakness of ureteral wall, where penetrate the vessels.


The effect of a synthetic polypeptide, threonyl-prolyl-arginyl-lysine, on ovulation in the squirrel monkey (Saimiri sciureus) and hamster.

Synthetic TPAL, a polypeptide reported to occur naturally in hamster zygotes, was tested at doses of 3 or 18 microgram/day for antiovulatory activity in cyclic hamsters (for 4 days) and squirrel monkeys (for 5 days) induced to ovulate. The TPAL treatment did not alter the ovulatory response in hamsters or monkeys.


A positive feedback effect of oestradiol on LH release in the male marmoset monkey, Callithrix jacchus.

Subcutaneous injections of oestradiol benzoate in oil, resulting in a sustained elevation of circulating oestradiol levels, induced an initial suppression of LH secretion, followed by a positive discharge of LH in castrated male and female and in intact male marmosets. Oestrogen-induced LH release (producing maximum LH concentrations 24 h after the injection) was observed in 75% of castrated males and females. A positive discharge of LH occurred in 50% of intact males 28-36 h after oestrogen administration.


Semisynthetic aminoglycoside antibacterials. 6. Synthesis of sisomicin, Antibiotic G-52, and novel 6'-substituted analogues of sisomicin from aminoglycoside 66-40C.

The discovery of aminoglycoside 66-40C, a novel dimeric, unsaturated imine produced by Micromonospora inyoensis, afforded a versatile intermediate for the synthesis of a variety of sisomicin analogues modified at the 6' position. The conversion of 66-40C into sisomicin, antibiotic G-52, and a series of novel 6'-substituted analogues of sisomicin is described, and the biological activity of the products is discussed.


Synthesis and analgetic activity of 1,2,3,4,5,6-hexahydro-1,6-methano-3-benzozocines.

1,2,3,4,5,6-Hexahydro-1,6-methano-3-benzazocine (1) has been synthesized via a four-step sequence from benzo-norbornadiene. This compound and its N-methyl derivative are more active than codeine in the mouse hot-plate antinociceptive assay and will not suppor morphine dependence in Rhesus monkeys.


The specificity of agglutination reactions of Pseudomonas aeruginosa with O antisera.

Polyagglutinable (PA) strains of Pseudomonas aeruginosa are agglutinated by more than one of the antisera prepared against antigenically unrelated O serogroups. They form c. 4% of strains--other than isolates from cystic-fibrosis units--submitted to us for typing. We were able to allocate 80% of PA strains to a single O serogroup by agglutination with typing sera that had been absorbed with the polyagglutinable strain SMC-247, or by co-agglutination tests with protein A-containing staphylococci coated with immunoglobulin from unabsorbed sera. Similar results were obtained by precipitation tests with crude bacterial extracts and unabsorbed sera, but these tests were less sensitive and less specific. Evidence is presented that PA antigen is a heat-stable cell constituent distinct from the O antigen. In rabbit antisera, anti-PA antibody is exclusively of the IgM class, but O antibody of both IgM and IgG classes is present.


p-nitrophenylglycerol--a superior antiswarming agent for isolating and identifying pathogens from clinical material.

The antiswarming agent p-nitrophenylglycerol (PNPG) has been found invaluable for the recognition and isolation of pathogenic bacteria from specimens contaminated with swarming strains of Proteus spp. PNPG is cheap, stable and non-toxic. All strains tested grew well in its presence and produced characteristic morphology. PNPG was without effect on the results of a variety of identification tests performed directly on colonies from media containing PNPG. This permitted identification without the need for further subculture.


Cyanide removal of gold from SEM specimens.

The cyanide method is highly recommended for the purposes of the removal of gold coatings from SEM specimens prior to (a) further treatment before further SEM, and (b) staining and mounting for LM.


Induction of monooxygenases in rhesus monkeys by 3-methylcholanthrene: metabolism and mutagenic activation of N-2-acetylaminofluorene and benzo(a)pyrene.

Induction of content and activity of hepatic and lung cytochrome P450-dependent monooxygenases by 3-methylcholanthrene (MCA) was studied in male rhesus monkeys. Hepatic microsomes were concomitantly studied for mutagenic activation of N-2-acetylaminofluorene (AAF) as well as with the use of sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Hepatic N-hydroxylation of AAF was increased sixfold to sevenfold after MCA treatment; aryl hydrocarbon hydroxylase was similarly increased in both liver and lung. The hepatic cytochrome P450 content was increased by 50%, and the Soret maximum in the CO-hemoprotein complex was shifted to the blue by 2 nm after MCA induction. Electrophoresis of hepatic microsomes from MCA-treated monkeys showed an increase in a polypeptide band of 54,000 molecular weight. Mutagenic activation of AAF by liver microsomes from untreated rhesus monkeys was low but was increased 40-fold after MCA treatment. No effect was observed on the mutagenic activation of N-hydroxy-2-acetylaminofluorene by liver microsomes after MCA treatment. Rhesus monkeys may provide a model for evaluation of the role of polycyclic hydrocarbon induction in chemically caused cancer and toxicity in primates.


Multiple autotransplantation of rat mammary induced by 7,12-dimethylbenz[a]anthracene: brief communication.

Mammary tumors induced in outbred Sprague-Dawley rats by 7,12-dimethylbenz]a]anthracene were excised, cut into 1- to 2-mm3 pieces, and then autotransplanted sc along the mammary line at six sites. Following an average period of 20--30 days, these autografts grew to approximately 2 cm in diameter in 32 of 48 rats (67%). Autografts in the other 33% of the rats remained dormant. Mammary tumors transplanted into allogeneic hosts failed to grow. Tumors derived from autotransplantation were indistinguishable from their primary tumors with respect to their histologic features, the nature of hormone dependency, the content of estrogen receptors, and their ability to incorporate [3H]leucine. Furthermore, autotransplanted tumors derived from a single primary tumor varied little with regard to the preceding parameters; thus they provided an opportunity for serial sampling of individual tumors for repeated morphologic and biochemical evaluations.


Lactate dehydrogenase activity and isoenzyme pattern in lungs, erythrocytes, and plasma of ozone-exposed rats and monkeys.

The effect of low levels of ozone exposure (8 hr daily for 7 days) on lactate dehydrogenase (LDH) activity and isoenzyme pattern was studied in the lungs, plasma, and erythrocytes of rats and monkeys. Exposure to 0.8 or 0.5 ppm ozone resulted in a significant increase in total LDH activvity in the lungs but not in the plasma and erythrocytes of rats. The LDH-5 fraction of the LDH isoenzymes was significantly decreased in terms of percentage distribution in the lungs and plasma of rats exposed to 0.8 ppm ozone, whereas the LDH-4 in the lungs and plasma and LDH-3 in the lungs were increased. At 0.5 ppm ozone, the isoenzyme distribution was not significantly changed in lungs, plasma, or erythrocytes. In monkeys, no significant change in either total LDH activity or isoenzyme pattern was observed in lungs, plasma, or erythrocytes after exposure to 0.8 ppm ozone.


Long-term effects of single and combined doses of DDT and PCB on drug-metabolizing enzymes in rat liver.

Two common environmental pollutants, DDT and a polychlorinated biphenyl (PCB) mixture (Clophen A-50), were administered ip to rats in discrete single doses (160 and 100 mg/kg, respectively) and in combination. All the enzyme activities studied were enhanced by DDT and PCB. the overall drug hydroxylation reactions, and their components, achieved maximal induction in 1 wk. The cytochrome P-450 content of microsomes was increased nearly fourfold and NADPH-cytochrome c reductase activity was enhanced twofold by both compounds. p-Nitroanisole O-demethylase was increased sevenfold by PCB and fourfold by DDT, aryl hydrocarbon hydroxylase threefold by PCB and 1.7-fold by DDT. After 2 wk the activities began to decline. Distinct increases in enzyme activities were still detectable 1 month after a single dose. Epoxide hydratase and UDPglucuronosyltransferase activities were also enhanced in 1 wk (epoxide hydratase 2.5-fold by both compounds, UDPglucuronosyltransferase tenfold by PCB in trypsin-activated microsomes but only threefold by DDT). The disappearance of induction in epoxide hydratase was slower than in the monooxygenases, and UDPglucuronosyltransferase still showed a trend toward increased activity 4 wk after the administration. The DDT-enhanced UDPglucuronosyltransferase activity slightly returned toward the control level. Glutathione S-transferase differed from the microsomal enzymes in that it was already elevated 1 day after the administration of both DDT and PCB. Its activity was only doubled, but the increased activity remained at almost the same level through the whole 1 month period.


Incorporation of (3H) lysine into the brain and spinal cord of the cebus (Cebus appella) and rhesus (Macacca mulata) monkeys.

The relative amounts of incorporation of [3H] lysine into various brain areas and segments of the spinal cord in adult Cebus and Rhesus monkeys were studied. One hour after subcutaneous injection of [3H] lysine, animals were sacrificed and the brain and spinal cord samples taken and processed for [3H] lysine incorporation into protein (Lowry et al., 1951; Wells and Bernstein, 1976). Analysis revealed marked differences in metabolism among brain areas within each species of monkey as well as across species. Cortical structures were generally higher in radioactive uptake than subcortical structures in both species. The Cebus monkey had the greatest variation among brain areas, particularyly between cortical regions, whereas the Rhesus monkey did not show cortical variation in uptake. The ratio of incorporated to nonincorporated lysine in the brain and spinal cord was higher in the Rhesus than the Cebus monkey. Both species showed a rostrocaudal gradient of radioactivity following [3H] lysine incorporation into the spinal cord. Considering weight, size, and availability differences, the metabolism of [3H] lysine appears to differ by brain areas and spinal cord segments between species.


Effects of Cannabis sativa on ultrastructure of the synapse in monkey brain.

Electron microscopic study of brain tissues of monkeys chronically exposed to psychoactive Cannabis showed widening of the synaptic cleft, electron opaque material in the cleft and in pre- and postsynaptic regions, and some "clumping" of synaptic vesicles. In contrast, tissues of control monkeys showed no ultrastructural changes.


Adsorption of cyanophage AS-1 to unicellular cyanobacteria and isolation of receptor material from Anacystis nidulans.

Cells of unicellular cyanobacteria of typological group Ia, containing approximately 50 mol% guanine + cytosine (G+C) in their DNA (R. Y. Stanier, R. Kunisawa, M. Mandel, and G. Cohen-Bazire, Bacteriol. Rev. 35:171-205, 1971), were susceptible to infection by the cyanophage AS-1. Cyanobacteria of the same typological group, containing approximately 65 mol% G+C in their DNA, did not adsorb the cyanophage AS-1 or adsorbed it at a low rate. AS-1 was not propagated by any of the investigated strains with a high G+C content in their DNA. However, cells of strains 6907 and 6911 were lysed by cyanophage AS-1. A comparison of the host range of this phage with the lipopolysaccharide composition of host and non-host cell walls suggests that lipopolysaccharides are involved in the adsorption process. About 8 microgram of lipopolysaccharide per ml from host strains inactivated 50% of the particles of a solution containing 100 PFU/ml after 60 min of incubation at 30 degrees C. Material with receptor activity was extracted from the host strain Anacystis nidulans KM. The extract was purified of glycolipids and pigments, and a fraction showing receptor activity was isolated. This fraction contained three polypeptides of molecular weights between 54,000 and 64,000. Heat and protease treatment of whole cells and of isolated receptor material decreased the receptor activity. The fluorescence intensity of A. nidulans cells labeled with 1-anilino-8-naphthalene sulfonate was increased when AS-1 was adsorbed to these cells. The participation of lipopolysaccharides and proteins in the formation of the receptor complex is discussed.


Transcriptional specificity of a multisubunit RNA polymerase induced by Bacillus subtilis bacteriophage PBS2.

Bacillus subtilis phage PBS2 induced the synthesis of two temporally defined categories of phage-specified transcripts. The transcription of phage "early" genes was induced almost immediately after infection; this RNA synthesis did not require phage protein synthesis. Phage "late" gene transcription, on the other hand, was induced at an intermediate time in the lytic cycle; this RNA synthesis required the production of phage proteins. Both classes of transcription were resistant to the drug rifampin and, therefore, apparently did not require the rifampin-sensitive component of the host RNA polymerase. A rifampin-resistant, DNA-dependent RNA polymerase was purified from bacteria infected with PBS2. The highly purified phage polymerase consisted of five distinct subunits that remained associated during zone centrifugation, isoelectric focusing, and disc gel electrophoresis. The synthesis of each of the five polypeptides was induced at an intermediate time in the phage lytic cycle. As judged by hybridization competition, hybridization to DNA restriction fragments, and RNA-RNA annealing, the phage-induced RNA polymerase preferentially and asymmetrically transcribed PBS2 late genes in vitro. These findings suggest that the PBS2 RNA polymerase controls the expression of phage genes late in the lytic cycle.


The effect of intravesical thio-tepa on normal and tumor urothelium.

Intravesical thio-tepa instillations for 55 patients with superficial transitional cell carcinoma were evaluated prospectively in the treatment of residual disease (therapeutic) and the prevention of recurrence (prophylactic). The over-all response rate to therapeutic thio-tepa was 56 per cent, with toxicity observed in 26 per cent of the cases. Patients with microscopic residual tumor showed better response and less toxicity than those with gross residual tumor. Prophylactic thio-tepa had no effect in lowering recurrence rates. Examination of pre-treatment and post-treatment biopsies showed no worsening of atypia and no correlation between improvement in atypia and gross tumor response. A charcteristic post-treatment thio-tepa effect was observed in non-tumorous urothelium but not in tumor biopsies. The differentiation of such thio-tepa effect cells from pre-malignant atypical cells is discussed.


Immunologic staging of prostatic carcinoma: three years of experience.

The immunological specificity of prostatic acid phosphatase has been demonstrated previously in the laboratory and used in a small number of patients. Our findings in 96 patients using gel immunodiffusion with an acid phosphatase antiserum in the study of bone marrow aspirates and lymph nodes are reported.


Nosocomial meningitis and bacteremia due to contaminated amphotericin B.

Nosocomial Gram-negative bacillary meningitis and bacteremia occurred in a patient who was receiving intrathecal and intravenous amphotericin B. An epidemiologic investigation found the amphotericin B to be contaminated with Enterobacter agglomerans, Pseudomonas fluorescens, and P aeruginosa. These contaminants were traced to a lot ot sodium phosphate buffer that was added to all intrathecal and intravenous amphotericin B preparations. The phosphate buffer underwent prolonged storage at room temperature and was not subject to terminal sterilization nor sterility testing. This parenteral admixture prepared in the hospital is now steam autoclaved and sterility tested before use.


Expected hemotherapy in elective surgery. A follow-up.

Preoperative cross match guidelines were developed from a review of elective surgical hemotherapy. Six elective surgical procedures were studied prospectively to determine the effectiveness of the recommendations. All six procedures demonstrated a substantial reduction in preoperative cross match ordering during the study period, although for only one procedure was the guidelines' recommendation followed strictly. For those procedures that rarely required hemotherapy, an ABO-Rh typing and an antibody detection test were recommended as a preoperative study without a cross match. Through education of the clinical staff, successful establishment of these guidelines can lead to substantial monetary savings, reduced blood outdating, and a decreased blood bank workload with a more appropriate allocation of the technologists' time and effort.


Formulating priorities for quality assurance activity. Description of a method and its application.

Quality assurance activity seems to have had little documented impact in terms of improving patient health or reducing care costs. One reason may be the lack of a practical and effective decision process for selecting priority areas where improvement of health or any other target outcome will most likely be achieved. This article describes a structured procedure for meeting this need. In addition, results of 14 years of quality assurance experience with structured and nonstructured topic selection procedures in 23 multispecialty group clinics and their associated hospitals are briefly reviewed. On the basis of this experience it is suggested that this priority method is both feasible and practical and can be recommended for application to most quality assurance systems. It is especially suited for planning medical care evaluation studies of the Professional Standards Review Organizations or the performance evaluation projects of the Joint Commission on the Accreditation of Hospitals.


Habits and dietary pattern of cases of carcinoma of the oral cavity and oropharynx.

In 1192 biopsy proven cases of carcinoma of the oral cavity and oropharynx information was collected on the use of various chewing, smoking, and eating habits. The quantity used and the duration of use were also recorded. Information was also collected on the frequency of use of cereals, dairy products, meat, fish, fruits and vegetables. The information from carcinoma cases was compared with that from apparently healthy individuals.


Aflatoxin and liver cancer in Karachi, a preliminary survey.

A cause and effect relationship of Aflatoxin and liver cancer is well established. The incidence of liver cancer is 3.6% in Karachi (Zuberi et al., 1976) and Aflatoxins have been detected in 43% of 28 samples of food analysed in this study. The long term ingestion of aflatoxin with food may have some effect on the existing pattern of liver cancer in Karachi.


Patients with acute poisoning seen in the department of intensive care-Jinnah Postgraduate Medical Centre, Karachi.

A review of 157 cases of acute poisoning admitted to the intensive care unit shows that poisoning was responsible for 40 15% of the cases treated at this unit between 15th January to 31st December, 1976. Inspite of limited facilities the over all mortality from acute poisoning was 9.5% while that due to organo-phosphorus insecticides alone was 7%. A considerable infrequency of organophosphorus insecticides poisoning was observed in developed countries when their data was compared with the present series.


Organo-phosphorus insecticide poisoning-review of 53 cases.

A review of 53 cases of organo-phosphorus insecticide poisoning admitted to the department of intensive care at the Jinnah Postgraduate Medical Centre between 15th January to 31st December, 1976 has shown that these insecticides form a majority of the causes of acute poisoning and are associated with a comparatively high mortality rate. Out of a total of 157 cases of acute poisoning admitted to the department, 34% were cases of organo-phosphorus insecticide poisoning forming 13 5% of the total admissions. These insecticides being easily accessible, are frequently used as suicidal agents specially by uneducated housewives. They are highly toxic and their management is very complicated. Of a total of 15 deaths from poisoning, 11 (73%) were due to this cause. This being such a common mode of poisoning in our country, we have presented here our experience of the clinical features and management of these cases.


Trace element analysis: a diagnostic tool.

Human body continuously assimilates a variety of elements from the environment and the concentration of these elements in blood is regulated by means of various homeostatic mechanisms. Some of the elements, though present in very small amounts perform highly specialized functions in initiating many biochemical reactions. These elements, known as essential trace elements, are closely related to human diseases as their deficiency or excess induces physiological changes. Many diseases such as hypertension, atherosclerosis, diabetes etc are related to trace element imbalance. Therefore the measurement of trace elements in body fluids and tissues can be effectively employed for diagnostic tests.


Restoration of the infectivity of Rickettsia tsutsugamushi to susceptible animals by passage in athymic nude mice.

Restoration of the infectivity of three prototypes of Rickettsia tsutsugamushi (i.e., Gillium, Karp and Kato strains) was examined to obtain each antiserum for the diagnosis of rickettsiosis and to prepare target antigens. When successively passed in embryonated yolk sac, rickettsiae gradually lost their infectivity to susceptible animals; after several passages in mice only Kato strain retained infectivity and others became completely avirulent. When these strains were passaged in athymic nude mice only once, they restored even stronger infectivity than original ones to other animals. The infectivity was retained in further passages in BALB/c, DDD mice or in guinea pigs. Thus excellent strain specific antisera were produced in guinea pigs, enabling defined diagnosis of rickettsial infection. Furthermore, successful restoration of the infectivity is expected to contribute to the studies on pathogenesis of rickettsiosis.


Infection protective property of the common antigen (OEP) of Pseudomonas aeruginosa and its chemical composition.

The protein moiety (OEP) of the endotoxin of Pseudomonas aeruginosa is a proteinaceous compound composed of possessing 77% protein and a small amount of lipid and sugar. The antigen has been proved to possess a common protective property against P. aeruginosa infections regardless of serotype and also to possess an antigen common to all the serotype strains of P. aeruginosa. When OEP was subjected to protease digestion, (protease-treated OEP) the protein content was reduced to 17% without a change in the lipid and sugar composition. The protease-treated OEP was no longer found to possess a serologically common antigen and was also found to have lost the common protective property against infections due to all the serotypes of P. aeruginosa. In conclusion, a serologically common antigen (OEP) as well as a common infection protective property was found to reside in a protein portion of the antigen.


Encephalomyelitis induced by canine distemper virus in non-human primates.

A strain of canine distemper virus was shown to be highly neuro-virulent in non-human primates. Intracerebral inoculation induced in monkeys histological lesions of encephalomyelitis, i.e., degenerative changes consisting mainly of neuronal damage and inflammatory changes such as perivascular cuffings and glial proliferation, in wide areas in the brain and spinal cord. In one monkey observed for 70 days, lesions with a tendency of subacute sclerosing were also noticed. Immunosuppression with cyclophosphamide or antithymocyte serum was found to aggravate the clinical course and to modify the histological lesions in the central nervous system as well as the level of antibody response to the virus in cerebrospinal fluid. Possible application of distemper encephalomyelitis in monkeys as a primate model for analysis of the immune mechanism involved in paramyxovirus-induced encephalomyelitis was discussed.


Chloride-induced contraction in a skinned fiber of toad striated muscle.

Cl- -induced contraction was studied in a skinned fiber dissected from toad sartorius muscle. When K-methanesulphonate in a relaxing solution was replaced isotonically with KCl, the skinned fiber contracted due to 'depolarization'-induced Ca++ release on the SR. Contraction was induced by superfusing the fiber with solutions containing different concentrations of Cl-. In the presence of 2 mM EGTA, the Cl- -induced contraction developed when the relaxing solution was replaced by a Cl- solution which contained up to 20 mM Cl-, while contraction started at 15 mM-Cl- in the presence of 0.5 mM EGTA. The peak tension of the Cl- -induced contraction depended on Cl- concentration without obeying the all-or-none law. When the fiber was treated with 1.5 mM caffeine, the threshold for the Cl- -induced contraction was significantly lowered. In spite of previous application of Cl- at different concentrations below 80 mM, the skinned fiber contracted by sudden increase in Cl- concentration, and the tension versus Cl- concentration curve revealed almost the same shape as that in a fiber equilibrated with Cl- -free solution. This suggests that the SR membrane is repolarized after transient depolarization caused by elevation of Cl- concentration.


Time- and Na-dependent effects of Ca depletion on potassium contracture in frog twitch muscle fiber.

The effect of the extracellular Ca depletion on potassium contracture was investigated in single fibers isolated from frog semitendinosus muscle mainly in relation to its time and Na dependency. The shortening of plateau duration and the increase in the rate of relaxation of the potassium contracture appeared within 3--5 sec and 15 sec, respectively, after the fiber was immersed in Ca-free Na Ringer solution containing 1 mM ethylene glycol bis(beta-aminoethyl ether)-N,N'-tetraacetic acid (EGTA) (EGTA-Na Ringer solution) or Ca-free choline Ringer solution containing 1 mM (EGTA (EGTA-choline Ringer solution). These effects were independent of the presence or absence of extracellular Na. In EGTA-Na Ringer solution, the potassium contracture tension was inhibited only by about 20% after 20--90 min and was abolished after 120 min. The inhibition of the peak tension was accelerated by the depletion of extracellular Na; in EGTA-choline Ringer solution, the tension was gradually inhibited by about 20% during the first 7 min and abolished after 10--12 min. When the peak tension of potassium contracture was abolished in EGTA-choline Ringer solution, the depolarization by Ca depletion was about 10 mV and the caffeine contracture was sufficiently produced. The results suggest that the inhibition of the potassium contracture tension in EGTA-choline Ringer solution is due to the dissociation of excitation-contraction coupling. On the basis of these results, an aspect of the inactivation of the potassium contracture was proposed.