Synthesis and activities of branched-chain aminoacyl-tRNA synthetases in threonine deaminase mutants of Escherichia coli.

Valyl-, isoleucyl-, and leucyl-tRNA synthetase activities were examined in an Escherichia coli K-12 strain that possessed a deletion of three genes of the ilv gene cluster, ilvD, A, and C, and in a strain with the same deletion that also carried the lambdadilvCB bacteriophage. It was observed that the branched-chain tRNA synthetase activities of both strains were considerably less than those of the normal strain during growth in unrestricted medium. Furthermore, during an isoleucine limitation, there was a further reduction in isoleucyl-tRNA synthetase activity and an absence of the isoleucine-mediated derepression of valyl-tRNA synthetase formation in both of these mutants, as compared with the normal strain. In addition, it was observed that these branched-chain synthetase activities were reduced in steady-state cultures of several ilvA point mutants. However, upon the introduction of the ilv operon to these ilvA mutants by use of lambda bacteriophage, there was a specific increase in the branched-chain synthetase activities to levels comparable to those of the normal strain. These results support our previous findings that the stability and repression control of synthesis of these synthetases require some product(s) missing in the ilvDAC deletion strain and strongly suggest this component is some form of the ilvA gene product, threonine deaminase.


Apparent suicidal inactivation of DNA polymerase by adenosine 2',3'-riboepoxide 5'-triphosphate.

Adenosine 2',3'-riboepoxide 5'-triphosphate (epoxyATP) has been found to be a suicidal inactivator of DNA polymerase I from Escherichia coli by the following criteria. Inactivation is complete, is first order in enzyme activity, and shows saturation kinetics with an apparent KD of 30 +/- 10 micron for epoxy ATP. This KD is comparable to the KM of the substrate dATP. The t1/2 for inactivation is 1.3 min. Inactivation requires Mg2+ and the complementary template. The enzyme is protected by dATP but not by an excess of template. Gel filtration of the reaction mixture after inactivation with [3H]epoxy ATP results in the comigration of E. coli DNA polymerase I, the tritium-labeled inactivator, and the DNA template. The stoichiometry of binding approaches 1 mol of [3H]epoxy nucleotide per mol of inactivated enzyme. These results are consistent with the hypothesis that epoxy ATP initially serves as a substrate for the polymerase reaction, elongating the DNA chain by a nucleotidyl unit, and subsequently alkylates an essential base at the primer terminus binding site of the enzyme. Epoxy ATP also inactivates human and viral DNA polymerases but not E. coli RNA polymerase or rabbit muscle pyruvate kinase. Hence epoxy ATP may be a specific suicide reagent for DNA polymerases.


Structural analysis of the tRNA1Tyr gene of Escherichia coli. A 178 base pair sequence that is repeated 3.14 times.

The distal region of the tRNA1Tyr gene has been sequenced and found to have an unusual structure. It consists of a 178 base pair sequence that is repeated 3.14 times. The first repeat unit commences 19 base pairs before the end of the sequences encoding the mature tRNA, and these 19 base pairs are repeated faithfully at the beginning of each repeat unit. In the last fractional unit the repeated sequence extends only six base pairs beyond this 19 base pair sequence. Sequence information extends for 62 base pairs beyond the 3.14 repeating units, and no resemblance to the repeating sequence, or any other region of the tRNA1Tyr gene, is found. There are only 14 sites at which one of the repeats differs from the others; 11 of these are transitions, and the rest are transversions. The evolutionary implications of the differences are discussed. One of the differences, which occurs in the second repeat unit, corresponds to the location of the in vitro p-dependent transcription termination site. This is discussed along with other implications of the repeated structure.


Escherichia coli mutants completely deficient in adenosylmethionine decarboxylase and in spermidine biosynthesis.

Mutants of Escherichia coli deficient in adenosylmethionine decarboxylase, an enzyme in the biosynthetic pathway for spermidine, were isolated after mutagenesis of E. coli K 12 with N-methyl-N-nitro-N-nitrosoguanidine or with the bacteriophage Mu. The mutated gene, designated speD, is at 2.7 min on the E. coli chromosome map. In several of the mutants resulting from Mu insertion both adenosylmethionine decarboxylase activity and spermidine were undetectable. The absence of spermidine from speD strains proves the essential role of adenosylmethionine decarboxylase in the biosynthetic pathway for spermidine. Despite the complete absence of spermidine, these mutants grew at 75% of the wild type rate.


The shape of L-arabinose isomerase from Escherichia coli.

L-Arabinose isomerase, EC 5.3.1.4, catalyzes the conversion of L-arabinose to L-ribulose, the first step in the catabolism of L-arabinose by Escherichia coli B/r. Patrick and Lee (1969) J. Biol. Chem. 244, 4277--4283) demonstrated that native L-arabinose isomerase is composed of six identical subunits of approximately Mr = 60,000. In this paper we describe an electron microscopy study of the arrangement of the six identical subunits. The isomerase is seen in two distinctly different orientations. The first has three subunits visible, with a 3-fold axis of symmetry, corresponding to a face-on view of two stacked, eclipsed trimers. The second orientation is rectangular in shape with 2-fold symmetry; suggesting a side-on view of the stacked trimers. The six identical subunits are thus arranged with D3 symmetry as in a trigonal prism. Measurements were made on the maximum profile of the three 2-fold axes of symmetry of the face-on orientations, and of both the long and short dimensions of the side-on orientation. The best estimate for the maximum profile of the 2-fold axes of symmetry of the face-on view is 106 +/- 8 A, using glutamine synthetase as an internal size standard. Measurements from micrographs of the isomerase alone, using an external magnification calibration, give the following results: for the maximum profile of the three 2-fold axes of symmetry of the face-on view, 132 +/- 7 A; for the long axis of the side-on view, 136 +/- 10 A; and for the short axis, 105 +/- 6 A. These measurements are consisting with the interpretation of the profiles as representing two different orientations of the L-arabinose isomerase.


Assembly of the mitochondrial membrane system. Genetic complementation of mit- mutations in mitochondrial DNA of Saccharomyces cerevisiae.

A method has been devised to test intergenic complementation of mutations in the mitochondrial DNA of Saccharomyces cerevisiae. The test is based on the observation that diploids issued from pairwise crosses of certain mit- mutants with deficiencies in cytochrome oxidase, or coenzyme QH2-cytochrome c reductase, acquire high levels of respiratory activity shortly after zygote formation. Under our experimental conditions neither biochemical complementation, interallelic complementation, nor recombination has been found to contribute to any significant extent toward the respiration measured in the diploids at early times. The test has been used to study the number of complementation groups represented by a large number of mit- mutants. Results of pairwise crosses of mutants in the oxi 1, oxi 2, oxi 3, cob 1, and cob 2 loci indicate that complementation occurs between the oxi and cob loci between different oxi loci but not between the two cob loci. The five loci have, therefore, been assigned to four different complementation groups.


The role of calcium in fusion of artificial vesicles.

Small phospholipid vesicles (liposomes) fuse upon calcium addition as demonstrated by electron microscopy, light absorbance increases, and mixing of original liposome contents within the boundaries of the fused liposome. The integrity of the fusion event is demonstrated by a novel assay based on the luminescence of firefly extract when mixed with ATP. Subsequent addition of valinomycin or the calcium ionophore A23187 leads to further fusion as shown by electron microscopy, light microscopy, and additional absorbance increase. Concomitant with this second absorbance increase is an increase in the amount of calcium that associates with the liposomes. This increased calcium association is more than can be accounted for by equilibration of 5 mM Ca2+ across the membrane and must indicate exposure of extra calcium binding sites. Binding of calcium to the inner side of the membrane may catalyze the second stage of liposome fusion.


Atlanto-axial arthrodesis by the wedge compression method.

Fifteen patients with atlanto-axial instability (secondary to os odontoideum in three, nonunion of an odontoid fracture in seven, acute odontoid fracture in three, and rheumatoid arthirtis in two) were treated by wedge compression arthrodesis of the atlanto-axial joint. One patient died at home eight weeks after fusion with the cause of death never established. Of the two patients with rheumatoid arthritis (ankylosing spondylitis), one had a non-union and in the other the posterior arch of the atlas fractured and the fusion had to be extended up to the occiput and down to the third cervical vertebra. The procedure is rarely indicated in patients with long-standing rheumatoid arthritis or severe osteopenia.


Atlanto-axial fusion for instability.

The problem of how best to treat a patient with instability of the atlanto-axial complex is still somewhat controversial. In this follow-up study of fifty-nine patients, nineteen were treated by a plaster jacket and brace; eleven, by single midline wiring and onlay bone grafts; and thirty, by four circumferential wires around the posterior elements of the axis and atlas with two bone grafts wedged between these elements on each side according to the method described by Brooks and Jenkins. (One patient had both types of fusion.) Although direct comparison of the results of treatment in three groups was not possible because of the many variables that may have influenced the results, the incidence of solid fusion was distinctly higher after the Brooks fusions despite less postoperative immobilization.


The protective value of a neurovascular island pedicle transfer in hands partially anesthetic due to ulnar denervation in leprosy.

Neurovascular skin island transfers were performed with the prime objective of protecting vulnerable anesthetic areas on the hands of patients with leprosy. After an average follow-up of eight years on sixteen patients, all had long-lasting protective benefits without further loss of tissue consequent to injury. At follow-up, two-point discrimination was less than ten millimeters in only one patient, more than twenty millimeters in ten, and indeterminate in five. Sensory misreference persisted in fourteen patients. Axon sprouting was evident in six but only over short distances (four to eight millimeters beyond the island). Compared with the intact side of the donor finger, nine of the sixteen transfers had lost some sensitivity but sensation was rated normal in one, nearly normal in six, and protective only in nine. The loss of sensation in the donor finger was less than expected and was not a problem. Scar contracture occurred in two donor and five recipient fingers, but this could be attributed to placement of the incision too far anteriorly, and hence was an unavoidable complication. Restoration of protective sensation to the ulnar border of the hand, whatever the cause of anesthesia, is considered extremely worth while.


Extra-articular low-velocity gunshot fractures of the radius and ulna.

Low-velocity gunshot fractures of the forearm are complex injuries and previously published studies have not emphasized the problems particular to these forearm lesions. Of the twenty-nine patients in this series, thirteen had peripheral nerve injuries, three had impending Voklmann's ischemia, and ten had delayed union or malunion of fractures after treatment by  closed methods. Only thirteen had none of these problems. Eight patients had long-term disability resulting either from permanent nerve damage with loss of sensation or weakness of grip, or from significant loss of motion following delayed union or malunion. Although external fixation was adequate for undisplaced fractures, delayed (seven to fourteen days) primary internal fixation after the initial phase of wound healing had proved benign gave superior results in displaced fractures.


Transposition of the tendo calcaneus for post-traumatic bone defects of the tibia.

Certain post-traumatic defects of the lower extremity present difficult problems in therapy and current methods of management do not offer satisfactory solutions. One such defect is an ulcerative lesion of the lower limb eroding the tibia and forming a chronic, rigid-walled cavity. We describe an operation in which the bone cavity is filled by viable tissue which acts as a bed for a skin graft. In chronic stasis ulcers it has been established that the tendo calcaneus will accept a skin graft, provided the paratenon is left undisturbred.


The role of spindle pole bodies and modified microtubule ends in the initiation of microtubule assembly in Saccharomyces cerevisiae.

The spindle poles of the budding yeast, Saccharomyces cerevisiae, have been removed from mitotic and meiotic cells by osmotic lysis of spheroplasts. The spindle pole bodies (SPBs)--diskoidal structures also termed 'spindle plaques'--have been analysed for their ability to potentiate the polymerization of microtubules in vitro. Free SPBs were completely deprived of any detectable native microtubules by incubation in the absence of added tubulin and were then challenged with chick neurotubulin, which had been rendered partially defective in self-initiation of repolymerization. Electron microscopy revealed that these SPBs served as foci for the initiation of microtubule polymerization in vitro. Because the attached microtubules elongated linearly with time but did not increase in numbers after the first stage of the reaction, it is apparent that there are a limited number of sites for initiation. The initiating potential of the SPBs was found to be inhibited by enzymic hydrolysis of protein but not of DNA. The microtubule end proximal to the site of initiation on the SPB is distinguished by a 'closed' appearance because of a terminal component which is continuous with the microtubule wall, whereas the distal end has the 'open' appearance characteristic of freely repolymerized neurotubules. SPBs which were partially purified on sucrose gradients retained their ability to initiate the assembly of microtubules with the same structural differentiation of their ends. The occurrence of closed proximal ends on native yeast microtubules suggests that closed ends may play a role in the initiation of microtubule polymerization in vivo, as well as in vitro.


Intracellular motility of mitochondria: role of the inner compartment in migration and shape changes of mitochondria in XTH-cells.

Mitochondrial movements have been followed by phase-contrast microscopy in living XTH-cells (Xenopus laevis tadpole-heart cells) in tissue culture. The same organelles have been viewed subsequently in electron micrographs. Locomotion of mitochondria proceeds at velocities up to 100 micrometer/min. Formation of branches of mitochondria and other shape changes may occur with the same speed. Mitochondrial motility can be classified into 4 types: (I) Alternating extension and contraction at the two ends of rod-shaped mitochondria. (2) Lateral branching. (3) Alternate stretching and contraction of arbitrary parts of a mitochondrion amounting to a kind of peristaltic action. (4) Transverse wave propagation along the organelle. Types I to 3 can be reduced to the same underlying principle; they cause locomotion. Formation of mitochondrial extensions is due to elongation of cristae. The observations are discussed in terms of 4 specific proposals. (I) Intracellular locomotion of mitochondria is caused by local enlargements and contractions of the organelles. (2) The shape changes are correlated with alterations in the arrangement of the cristae. (3) Such arrangements are not associated with overall swelling or shrinkage of the mitochondrion; they are local features. (4) Estimates of the time required for rearrangement of the inner compartment amount to less than 0.3 s for single crista arrangements during the fastest shape changes, and less than 1-3 s during slower alterations. This high velocity is in good accord with the hypothesis of energy conservation by conformational events during oxidative phosphorylation.


[Breakdown of the anastomosis after colectomy. The causes and prevention (author's transl)].

Over a period of 3 years the authors have carried out 184 colectomies with immediate restoration of continuity. In 25 of these operated patients the sutures broke down, i.e. 13.5% and 8 died, i.e. one patient out of 3. In the light of this series, the authors studied factors favouring breakdown, whether relative to the patient (age, sex, pathology, nature of the lesion) or the technic followed, i.e. in fact the surgeon (extent of the colonic resection, type and mode of anastomosis, modes of suture and drainage). Were studied also, in prevention, the first or contemporaneous construction of a colostomy, which according to the authors totally prevents breakdown of the subjacent anastomosis, but has its own complications.


[Pancreatic transplantation. Implantation of auto and allografts of the pancreas into the gastric wall of the diabetic rabbit (author's transl)].

The author's studied pancreatic autotransplants and allotransplants in the gastric submucosa of the rabbit after removal of the pancreas. Pancreatctomy causes severe diabetes with hyperkaliemic acidosis which is always fatal with 72 hours. Auto or allotransplantation cures the diabetes within two to three days, and maintains normal glucose balance for about 60 days. Early death after transplantation occurred when the graft was totally ineffective. After transplantation, the blood sugar rose progressively towards the 15th day, then suddenly and definetely at the time of necrosis of the graft towards the 15th or 21st day. The follow up in some transplanted animals exceeded two months. The interest of the procedure resides in the implantation of a large critical mass (25 to 30% of the pancreas ) in a well-vascularised area, drained by the portal system, and permitting the digestive tract elimination of the pancreatic necrosis whether of vascular or immune origin.


Determination of the hemin requirement of Haemophilus ducreyi: evaluation of the porphyrin test and media used in the satellite growth test.

Gonococcal (GC) agar supplemented with glucose and glutamine was found to be superior to Eugonagar and Trypticase soy agar in demonstrating the hemin requirement of 23 strains of Haemophilus ducreyi by the satellite growth test. The porphyrin test confirmed the requirement for exogenous hematin. With the agar dilution technique, using supplemented GC agar, the hemin concentration required to initiate growth was 10 microgram/ml, and the optimal hemin concentration to produce growth equivalent to that on chocolate agar was between 200 and 500 microgram/ml. On GC agar with added glucose and glutamine, the lowest hemin concentration impregnated in paper disks able to initiate satellite growth was 50 microgram/ml. The hemin requirements of these H. ducreyi were much higher than that reported for other Haemophilus species.


Premarket evaluation of commercial toxoplasmosis indirect fluorescent-antibody reagents.

The quality of commercial toxoplasmosis indirect fluorescent-antibody reagents was evaluated over a 6-year period. Seven manufacturers voluntarily submitted their products for evaluation in the Center for Disease Control Premarket Evaluation Program. Each product was tested in accordance with the Center for Disease Control performance and labeling specifications and evaluation methods. Only 49% of all of the products tested met the Center for Disease Control requirements. Performance criteria are outlined, and suggestions are offered to assist laboratorians in obtaining toxoplasmosis indirect fluorescent-antibody reagents of high quality.


Electrical impedance measurements: rapid method for detecting and monitoring microorganisms.

A conceptually simple and east-to-use technique is described that uses continuous impedance measurements for automated monitoring of microbial growth and metabolism. The method has been applied to a wide range of microorganisms. Optical clarity is not required. The sensitivity and reproducibility of the method are demonstrated. The mechanism whereby microbial growth alters the impedance of the medium is discussed, as well as potential applications of the method to clinical microbiology.


Impedimetric screening for bacteriuria.

A rapid, automated instrumental procedure for distinguishing urine cultures containing greater than 10(5) organism per ml is described. The method is based upon the measurement of changes in impedance that take place as microorganisms alter the chemical composition of the medium. The time required to detect impedance change is inversely related to the initial concentration of microorganisms in the sample. By defining an impedance-positive culture as one that gives detectable impedance change within 2.6 h, 95.8% of 1,133 urine cultures tested were correctly classified as containing more than or fewer than 10(5) organisms per ml. Selection of a longer detection time decreases false negative results at the cost of increased false positive results. Impedance screening is compared with screening data reported in the literature using adenosine-5'-triphosphate detection, microcalorimetry, electrochemical measurements, and optical microscopy.


L-Arabinose-ornithine-Irgasan medium for differentiating Serratia species.

A semisolid medium (designated Serratia differentiation medium) containing L-arabinose, ornithine, and selective inhibitor was used to differentiate three clinically encountered Serratia species. The inhibitor, Irgasan DP-300, was incorporated to eliminate false-positive reactions from most remaining Enterobacteriaceae. The suspected Serratia colony was inoculated as a stab into the medium. Serratia marcescens was indicated by a change in color from olive to purple following 18 h of incubation, whereas S. rubidaea (not listed in Bergey's Manual of Determinative Bacteriology) was indicated by a change to bright yellow. S. liquefaciens (described in Bergey's Manual of Determinative Bacteriology [8th ed., 1974] as Enterobacter liquefaciens) produced a small purple band at the top of the medium and a yellow or yellow-green butt. Absence of growth and color change following incubation indicates that the suspected colony is a non-Serratia. Thirty-six Serratia strains and 97 other Enterobacteriaceae and Pseudomonadaceae strains were tested. Two strains of the non-Serratia Enterobacteriaceae (one each of Citrobacter freundii and Proteus morganii) and two strains of Pseudomonas aeruginosa produced a color change in the medium. All of the Serratia strains tested were correctly identified using this medium, while 96% of the other species tested were inhibited.


Simplified microscopy for rapid detection of significant bacteriuria in random urine specimens.

Simplified urine microscopy, nitrite testing, and dipstick culture were compared with urine loop streak culture colony counts in 219 random voided specimens to determine the accuracy of the three rapid screening techniques. Nitrite testing resulted in 65% false negative results, which could not be significantly improved by incubation at 37 degrees C but which could be improved by adding nitrate substrate before incubation. Dipstick culture could not be quantitated until after 18 h of incubation. A new, simplified microscopy technique, using unspun, unstained urine, resulted in 4% false negative results and 4% false positive results in specimens containing over 10(5) organisms per ml and was the best method Centrifuges, Gram staining reagents, and counting chambers are not necessary for accurate microscopic screening of random urine specimens for the presence of bacteriuria by this technique, and the results are immediately available.


Evaluation of an automated agar plate streaker.

An automated agar plate streaker was evaluated. The Autostreaker mechanizes the agar plate streaking process by providing storage for plates, labeling and streaking one or more plates for either isolation or quantitation, and stacking in one of several racks for subsequent incubation. Results showed the Autostreaker to produce agar plates with well-separated colonies and accurate colony counts. A total of 1,930 clinical specimens were processed either in parallel with manual methods or solely by the Autostreaker. Technologist acceptance of machine-streaked plates was outstanding.


Use of the Minitek system for biotyping Haemophilus species.

The Minitek system was used for characterization and identification of Haemophilus. The system was simple to use and provided rapid and clear-cut test results.


Atypical isolate of Cryptococcus neoformans cultured from sputum of a patient with pulmonary cancer and blastomycosis.

Cryptococcus neoformans was isolated repeatedly from a patient with epider-moid carcinoma and pulmonary blastomycosis. The isolate was atypical in that it had only a minute capsule, caused persistent infection but no perceptible disease in mice, and initially appeared not to assimilate trehalose. Only after an incubation of 2 to 3 weeks did utilization of this substrate become apparent.


Quantitative recovery of spores from thermophilic spore papers.

A homogeniser (Stomacher 400) which does not damage bacteria has been used to produce an homogenate of spore papers, from which it was possible to make quantitative assessments of viable spores recoverable from such papers before and after exposure to sterilising procedures.


Performance of converted pressure cookers and two conventional jars for anaerobic bacterial culture.

The simple conversion of commercial pressure cookers into inexpensive anaerobic jars is described. These containers were shown to be as good as the small conventional BBL polycarbonate GasPak and large vented 150 gas-replacement jars when assessed by means of three biological indicators: Pseudomonas aeruginosa, Bacteroides melaninogenicus, and Bacteroides fragilis. Ps. aeruginosa seeded on Simmond's citrate agar was shown to be the most sensitive indicator of the three for traces of oxygen.


Survival of upper respiratory tract bacteria on cotton-wool swabs.

Plain, buffered cotton-wool swabs and albumen-coated cotton-wool swabs were prepared with known numbers of the following: Haemophilus influenzae, Streptococcus pneumoniae, Staphylococcus aureus, corynebacteria and Escherichia coli. Swabs were plated immediately (0 hours) after loading with a 0.1 ml standardised inoculum; each swab was plated out four times by streaking on both halves of two plates. At 0 hours recovery rates from first platings on solid media ranged from 3.3%-8.9%; from platings 1-4 ranges were 2.5%-5.7%. Both types of swabs gave similar results. The greater the delay in plating swabs at room temperature and 4 degrees C the lower was the recovery rate of H. influenzae and Strep. pneumoniae, though both species survived better on either kind of swab when these were held at 4 degrees C. There was a very small reduction in numbers of Staph. aureus from 24 to 48 hours but the recovery rate of corynebacteria was similar over the 48-hour period, at both room temperature and 4 degrees C. Twenty-four-hour counts of E. coli at room temperature were approximately five times greater than at 0 hours, and at 48 hours colonies were too numerous to count. At 4 degrees C, however, counts of E. coli fell during the 48-hour period.


Structure of the leech nerve cord: distribution of neurons and organization of fiber pathways.

The abdominal nerve cord of the leech Macrobdella decora was studied under the light and electron microscopes. The ganglionic cortex consists of six hemicone-shaped packets of neuronal perikarya and apical processes regularly assembled in bilaterally symmetric rows. The orderly projection of the apical processes into the hilum of the packets is also followed by an orderly distribution of their branches across the neuropile. This part of the ganglion is made of two symmetrical halves or hemineuropiles enclosing two types of nerve tissue: coarse and fine neuropiles. The coarse neuropile has seven longitudinal and four commissural tracts of fibers and a distinctively segregated synaptic zone. Nerve processes in this neuropile mostly proceed from the neurons in the ganglia and some are the branches of giant afferent axons. The fine neuropile includes several longitudinal tracts of fibers and a non-segregated synaptic zone. Most nerve processes in this neuropile are small afferent axons and some come from neurons in the ganglia. Bundles of axons in the connectives result by the orderly projection of the neuropile longitudinal tracts and together form fiber pathways connecting the synaptic zones of successive ganglia. Pathways of through-ganglia giant axons, linking the coarse neuropile synaptic zones, and of small axons, linking the fine neuropile synaptic zones, are described.


Transient acantholytic dermatosis with involvement of oral mucosa.

Histologic, immunologic and electron microscopic studies were performed in a patient with transient acantholytic dermatosis which involved oral mucosa. Hitologic and electron microscopic findings were almost identical in both cutaneous and mucous membrane lesions, and these were similar to pemphigus vulgaris; suprabasilar separation with acantholytic cells. Desmosome-desmosome complexes were separated without disruption of cell membranes and cytoplasmic structures were well-preserved without dyskeratosis. Lamina lucida, however, was often separated in mucous membrane lesions, in contrast to the normal lamina lucida in cutaneous lesions of such cases or in pemphigus vulgaris. Direct and indirect immunofluorescence studies for pemphigus were repeatedly negative. This study shows that transient acantholytic dermatosis may involve mucous membrane and may resemble pemphigus vulgaris histologically and ultrastructurally except for the widened lamina lucida of the mucous membrane.


Effects of alexidine.2HCL mouthwash on plaque and gingivitis after six months.

Two-hundred and fourteen subjects ranging in age from 22 to 63 years used either a mouthwash that contained 0.035% w/v alexidine.2HCL (2-ethylhexyl bisbiguanidine dihydrochloride) or a placebo twice daily on a doubleblind basis for six months in conjunction with routine oral hygiene procedures. The results indicated that alexidine.2HCL was safe and was statistically more effective in producing a clinically significant reduction in gingivitis than was the placebo.


Documentation of vasomotor dysfunction by indicator dilution curve analysis.

Indocyanine green dye dilution curves with abnormal contours have been recorded in four patients with acute and critical illnesses using standardized recording methods. The abnormal curves are characterized by a high and prolonged concentration of dye occurring after the peak concentration. This altered contour can be detected by use of routine mathematical processing since the derived hemodynamic parameters are affected by the altered contour. A physiologic basis for the altered curve contour is proposed.


Claspless chrome-cobalt transitional removable partial dentures.

Many of the acrylic resin and bent-wire temporary partial dentures in use today produce pathologic changes in the oral mucosa. A method is outlined for the use of chrome-cobalt, instead of acrylic resin, to produce temporary prostheses. The resulting temporary partial dentures are esthetically pleasing and biologically acceptable to the oral tissues.


An investigation of bonding systems for the polycarbonate crown restoration.

Tensile retentive strengths for six polycarbonate crown cementation systems were examined with use of extracted human teeth prepared with and without retentive grooves. From the results of this study, use of unfilled intermediary resins followed by polycarboxylate or zinc phosphate cements is recommended. Retentive grooves did not contribute to overall retention.


Sequencing and timing of treatment in the correction of dentofacial deformities in adult patients with clefts.

During the past six years, 107 patients with various dentofacial deformities secondary to primary clefts were treated in the cleft palate clinic. An overall treatment plan was formulated before any definitive secondary treatment was started. The sequencing and timing of the various aspects of secondary treatment were established to provide optimum esthetic, functional, and psychologic results.


Alcohol and nutrient intake of elderly men.

The effect of alcohol consumption on the intake of food nutrients was investigated in elderly residents in a Veterans Administration Domiciliary. When compared with a group of abstainers from the same institution, the imbibers ate fewer food calories (2,140 vs. 2,415 kcal per day) and had lower serum albumin levels (54 vs. 58 per cent total serum protein). The lower caloric intake extended to the three main energy sources (fat, protein, and carbohydrate) and occurred even though ample food was available. In view of the reduced nutrient intake and known impairment of the absorptive process by ethanol, institutionalized elderly alcoholics require special attention to maintain a satisfactory nutritional status.


Research in orthokeratology. Part VIII: results, conclusions and discussion of techniques.

In this paper, the results, discussions and conclusions concerning selected variables relative to orthokeratological effects and procedures are presented. The data, clinical observations and subsequent analyses suggest that corneal modification with contact lenses is not clearly understood. Hypothesis are offered to explain certain results obtained, to account for observed discrepancies and to collectively assemble a rational understanding for the effects of contact lenses on purposeful corneal modification.


Counts on viable Bacteroides fragilis: a modification of the microdroplet technique.

A modification of Sharpe's microdroplet counting technique is presented which permits viable counting of Bacteroides fragilis. Lysed horse blood was added to 0.1 soy digest agar droplets and 2% was found to be the optimal concentration which combined adequate intradroplet colony size with acceptable visibility on the counting screen of the Colworth Droplette machine.


Serological evidence of continuing infection of swine in Great Britain with an influenza A virus (H3N2).

Serum samples collected from swine and cattle in Great Britain at various times between July 1971 and July 1977 were examined by haemagglutination-inhibition or single radial haemolysis methods for evidence of infection with influenza A (H3N2) viruses. A small proportion of swine sera collected in each year reacted in the tests but there was no evidence of infection in cattle. The significance of the findings is discussed, with particular reference to the seasonal fluctuations in the prevalence of antibody in swine observed during the period of the study, and their possible relevance to influenzal events in the human population. None of the sera tested had antibody to swine influenza strains (HSw1N1).


Bowls and bacteria.

An episode of nosocomial infection with Klebsiella aerogenes in a surgical ward, in which six patients were infected, is described. The cause of the outbreak was identified as being contaminated washing-bowls. It is recommended that each patient should have his own bowl, which should be disinfected after use, dried and stored upside-down.


Clostridium botulinum in aquatic environments in Great Britain and Ireland.

Mud samples from aquatic environments in many parts of Great Britain and Ireland were collected, mainly in 1975 and 1976, and examined for Clostridium botulinum. The samples were taken from lakes, ponds, reservoirs, marshes, mudflats, streams, rivers and canals, and the sampling areas included a number of bird refuges and reserves. Of 554 samples 194 (35.0%) were positive and 167 (30.1%), 19 (3.4%), 6 (1.1%) and 15 (2.7%) contained types B, C, D and E respectively; 13 (2.3%) contained more than one type. Each type demonstrated was found in both fresh-and salt-water environments. Type B was widespread; types C, D and E were demonstrated in widely separated areas in England and Wales, and type C was found in both the north and south of Scotland. The results were compared with those reported earlier in respect of surveys in the London area, the Norfolk Boads and the Camargue (France).


A new assay for rapid measurement of MIF levels by 3H-labelled cells in liquid scintillation counting vials: statistical implications for the measurement of migration inhibition.

A new quantitative assay for migration inhibitory factor (MIF) employs 3H-labelled cultured mouse or human lymphoid cells migrating from capillary tubes. Capillaries filled with labelled cells are placed in liquid scintillation counting vials, along with the MIF-containing sample and are removed at the end of a five-hour incubation period. The residual, labelled cells which have migrated out of the tubes are solubilized and counted in a liquid scintillation counter. While cultured lymphoblast cells are routinely used in the assay, the method was checked against mouse and guinea pig peritoneal exudate cells in both the labelled cell technique and the conventional chamber assay. The assay is technically simple to perform and a useful tool for laboratory research purposes because of the short span of time needed to obtain the results. These advantages indicate a potential for automation and use of this assay in a clinical immunology laboratory. Statistical analysis of data from both assays demonstrated that the relative variation among replicates is lower in the labelled cell assay. The new assay also measured a significant difference between controls and MIF-containing samples when the migration index (MI) was greater than 80%. Criteria for significant inhibition of migration are discussed in regard to the use of analysis of variance (ANOVA) and other statistical procedures, and the inadequacy of a single measure, such as the MI, is discussed.


Use of staphylococcal protein A as an immunological reagent.

This brief review summarises the major uses of staphylococcal protein A in immunology. Protein A is covalently linked to the cell wall of most strains of Staphylococcus aureus, and binds immunoglobulin molecules with high affinity. The principal molecule bo-nd is IgG, although in many cases binding is restricted to certain IgG subclasses. Some IgM and IgA binds in certain species. This property allows rapid, simple and economical methods for the purification and analysis of immunoglobulins, and the fractionation of subclasses which are difficult to separate by other means. Fractionation on protein A affinity columns is a simple and efficient way of separating immunoglobulin F (ab) and F (ab')2 from Fc fragments. Intact staphylococci are useful as a solid phase adsorbent for isolating antigen-antibody complexes, membrane antigens and receptors, and to replace 'second antibody' in radioimmunoassay. Finally, protein A has proven useful for the study of antigens and receptors on the surface of intact cells, and for the detection of antibody-secreting cells. Thus, the use of protein A is now the method of choice for many preparative and analytical purposes in immunology.


Purification of human monocytes by continuous gradient sedimentation in ficoll.

A new method of obtaining purified human monocytes has been developed. The peripheral blood mononuclear leukocytes are isolated by centrifugation over Ficoll--Hypaque and then further purified by sedimentation over a linear 5--10% Ficoll density gradient. In ten experiments, the average purity obtained was 77.1% macrophages and the mean yield was 22.4% of the monocytes contained in the peripheral blood leukocytes. Viability of monocytes isolated by this technique exceeded 95%. The cells were phagocytic and responded to human migration inhibitory factor.


An improved method for isolation of granulocytes from peripheral blood.

A method is described by which granulocytes may be isolated from human peripheral blood. The yield of granulocytes using this technique (70.9%) is considerably greater than that obtained by alternative methods. The isolated cells are mainly composed of neutrophils, but the ratio of eosinophils to neutrophils is not significantly different from that of the peripheral blood. The cells are functionally comparable to buffy coat granulocytes with respect to phagocytic, candidacidal and NBT dye-reduction ability. Removal of erythrocytes from the final cells gives a granulocyte preparation useful for studies in which purity is essential.


A new plastic receptacle for solid phase immunoassays.

A new, simple, inexpensive plastic device for solid phase immunoassay, the through-passage receptacle (TPR), with a large ratio of surface area to volume, is described. It was substantially more efficient in non-specific protein adsorption and specific immunological binding than the conventional wells of microtiter plates. It was also easier to wash since washing was facilitated by a through-passage with a gravity flow. This device, which should be more conducive to automation than currently used receptacles, should increase the sensitivity and decrease the incubation time of the solid phase immunoassay technique.


New immunofluorescent antibody test for diagnosis of candidiasis using macrophages-engulfed antigens as substrate.

A new method for detection of antibodies to polysaccharidic antigens of Candida albicans is described. The method is based on the selective uptake of carbohydrate-rich antigens by mouse peritoneal macrophages, the antibodies being revealed by indirect immunofluorescence after incubation of fixed, antigen-containing macrophages with sera (MU/IFA test). This test shows greater sensitivity than counterimmunoelectrophoresis individuals. The combination of CIE using cytoplasmic antigens and MU/IFA using surface antigens and MU/IFA using surface antigens is useful in a considerable majority of cases for diagnosis of clinical infection.


A micro method for the estimation of killing and phagocytosis of Candida albicans by human leucocytes.

A micro method is described for quantitation of the candidacidal and phagocytic capacity of human neutrophils based on previous studies which showed that non-viable Candida albicans are differentially stained by methylene blue. The assay was performed on a microplate using small numbers of phagocytes (2 x 10(3) per well) and Candida albicans (2 x 10(3) or 8 x 10(3) per well) and both the phagocytic and microbicidal activity could be observed in situ within the phagocytic cell. The technique requires small volumes of blood (2 ml) and simple apparatus. Plates may be stored after fixation of stained preparations which makes it particularly suitable as an initial screen of phagocyte function for routine clinical use.


Immunochemical analysis of the monoclonal paraprotein in scleromyxedema.

The monoclonal paraprotein from a typical case of scleromyxedema was isolated and characterized. The isolated paraprotein was of the IgG-lambda class, with a molecular weight of approximately 110,000 daltons compared with 160,000 daltons for normal IgG. Immunochemical studies indicated that the paraprotein was an incomplete IgG molecule which was missing a significant antigenic portion of the Fd fragment.


Plastic embedding of cutaneous specimens for light microscopy using araldite epoxy resin.

A method is described using araldite epoxy resin for embedding intact 3-mm cutaneous punch biopsies for light microscopy. Sections are cut at 0.5-2.0 micron and stained with hematoxylin-eosin-phloxine. This procedure allows for preservation of cellular detail unobtainable with conventional formalin-fixed, paraffin embedded tissue.


Cefamandole and cefazolin in the therapy of complicated urinary tract infections.

The clinical efficacy and tolerance of cefamandole, a new cephalosporin antibiotic effective against indole-positive strains of Proteus, and cefazolin were studied after intramuscular administration of 500 mg of either of the two cephalosporins every 8 hr for seven days in a prospective, randomized study of 65 elderly male patients with complicated urinary tarct infections. Both antibiotics were effective in eradicating the infections, and there was no significant difference between the two groups in regard to tolerance and cure rate, as defined by a negative urine culture one week and four to six weeks following discontinuation of the treatment. Because of its broader antibacterial spectrum, cefamandole appears to represent an improvement over previously available cephalosporin antibiotics.


Cefamandole vs. procaine penicillin for treatment of pneumonia due to Streptococcus pneumoniae: a random trial.

The efficacy and safety of cefamandole nafate and penicillin G procaine suspension were compared in the treatment of pneumococcal pneumonia in hospitalized adults. One hundred thirteen patients with clinical and radiographic evidence of pneumococcal pneumonia were randomly assigned to receive 600,000 units of procaine penicillin intramuscularly every 12 hr or 500 mg of cefamandole intramuscularly every 6 hr. The two groups were comparable with regard to patient type and extent and severity of pneumonia. Alcohol abuse was a host factor in 31% of all patients in the trial. All strains of Streptococcus pneumoniae isolated were inhibited by less than or equal to 1.6 microgram of cefamandole/ml. Of 58 patients treated with cefamandole, 50 had a satisfactory response, as did 46 of the 55 patients treated with penicillin. Results of tests of liver function were abnormal (primarily, elevated levels of transaminase or alkaline phosphatase) in 38% of the entire group of patients and occurred with equal frequency in patients receiving cefamandole or penicillin. Side effects during therapy, including superinfection, occurred equally with either drug. In a random trial, cefamandole was as effective and safe as penicillin in the treatment of pneumococcal pneumonia in adults.


Efficacy and safety of cefamandole plus either gentamicin or tobramycin in therapy of severe gram-negative bacterial infections.

Thirty-one patients with severe gram-negative bacterial infections were treated successfully with a combination of cefamandole nafate plus gentamicin or tobramycin. The patients were divided into two treatment groups: group 1 received low-dose therapy (80--100 mg of cefamandole nafate/kg per 24 hr plus 3 mg of either gentamicin or tobramycin/kg per 24 hr), and group 2 patients, who had suspected bacteremia, received high-dose therapy (170 mg of cefamandole nafate/kg per 24 hr plus 5 mg of either gentamicin or tobramycin/kg per 24 hr). All of the patients were clinically and bacteriologically cured of their primary infections. All four episodes of bacteremia were cleared within 24 hr after therapy was initiated. There was a uniform decrease in the rate of creatinine clearance which was slightly greater in group 2 patients; however, all creatinine clearance values were within the normal range and actually improved during therapy. There was no difference between the clearance values of the tobramycin-treated patients and gentamicin-treated patients. A few transient abnormalities in results of liver function tests occurred during the study. In one patient whose serum was positive for hepatitis-associated antigen, the alkaline phosphatase, aspartate aminotransferase, and bilirubin values were elevated on admisssion of the patient to the hospital, increased fivefold during therapy, and decreased to the base-line admission values six days after therapy; however, it is difficult to establish that this reaction was antibiotic-induced hepatic toxicity.


Clinical and laboratory investigation of cefamandole therapy of infections in infants and children.

Cefamandole nafate was effective in the treatment of a variety of infections caused by Staphylococcus aureus, Streptococcus pyogenes group A, Streptococcus pneumoniae, and Haemophilus influenzae in infants and children. The infections included periorbital cellulitis and ethmoiditis, bacteremia, cellulitis, pneumonia, and lymphadenitis. In vitro, cefamandole was effective in inhibiting the growth of H. influenzae isolated from blood or cerebrospinal fluid of patients with meningitis or sepsis. In two patients rash developed and cefamandole was discontinued. Other significant adverse effects were not noted.


Antibacterial activity of cefamandole in vitro.

Cefamandole, a new cephalosporin derivative, was found to have a broad spectrum of antimicrobial activity against a cross-section of both gram-positive and gram-negative bacteria isolated clinically. Gram-positive cocci, except for Streptococcus faecalis, were extremely susceptible to cefamandole; penicillin G-resistant Staphylococcus aureus also was highly susceptible. Minimal bactericidal concentrations for gram-positive cocci approximated the minimal inhibitory concentrations. Strains of Haemophilus influenzae were very susceptible to the drug. Most strains of Escherichia coli, Klebsiella species, and Proteus species were inhibited by low concentrations of cefamandole, Salmonella typhi, including ampicillin- and chloramphenicol-resistant strains, was inhibited by low concentrations of cefamandole. Susceptible bacteria became increasingly resistant as the inoculum size was increased. Strains of Pseudomonas were resistant to cefamandole.


Differences between cephalothin and newer parenterally absorbed cephalosporins in vitro: a justification for separate disks.

The activities of cefamandole and cefoxitin in vitro were compared with that of cephalothin against staphylococci and gram-negative bacilli. Cephalothin was the most active agent against staphylococci. Cefamandole exhibited the greatest activity against the Enterobacteriaceae, with the exceptions of Serratia marcescens and indole-positive Proteeae, against which cefoxitin was the most active antibiotic. The activity of newer cephalosporins that are resistant to the beta-lactamases of gram-negative bacteria renders untenable the status of cephalothin as the class representative of the cephalosporins for susceptibility testing.


A comparative study of the activity of cefamandole and other cephalosporins and analysis of the beta-lactamase stability and synergy of cefamandole with aminoglycosides.

The antibacterial activity of cefamandole against 445 clinical isolates was investigated and compared with the activity of other known cephalosporins (cephalothin, cephaloridine, cephalexin, and cefazolin) and of two penicillins (ampicillin and carbenicillin). Cefamandole was the most active antibiotic against isolates of Citrobacter, Enterobacter, and Shigella, and its activity against Staphylococcus aureus, Bacteroides, and some members of the Enterobacteriaceae was comparable to that of the other antibiotics tested. The stability of cefamandole with respect to beta-lactamase was investigated and compared with that of cephalothin, cefazolin, and cephalexin. Cefamandole was stable with respect to the beta-lactamases of Enterobacter and some other members of the Enterobacteriaceae. No significant correlation was found between the antibacterial activity and the beta-lactamase stability of cefamandole, except with Enterobacter. The synergistic activity of cefamandole combined with gentamicin or amikacin was demonstrated by killing-curve techniques, isobolograms, and susceptibility data. Although 12%--46% of the isolates were synergistically inhibited by either combination, antagonism was not observed. No correlation between the hydrolysis of cefamandole by beta-lactamase and the synergistic activity of cefamandole combined with amikacin was demonstrated.


Pharmacokinetics of cephalosporins in patients with normal or reduced renal function.

The pharmacokinetic distribution of 10 cephalosporin compounds, cephalothin, cephaloridine, cephaloglycine, cephalexin, cefazolin, cephapirin, cephradine, cephacentrile, cefoxitin, and cefamandole, in patients with various degrees of renal function was reviewed. The mean serum half-life (t1/2) of each cephalosporin compound was calculated from reported data in patients grouped according to their degree of renal function, as determined by the rate of creatinine clearance. Mean t1/2 values in patients with normal renal function were compared with t1/2 values for the same cephalosporin in patients with moderate renal failure and severe renal azotemia. The effect of hemodialysis and peritoneal dialysis on the mean serum t1/2 of each cephalosporin was also determined. The mean serum t1/2 of each cephalosporin progressively increased as the rate of creatinine clearance decreased. Hemodialysis decreases the t1/2 of cephaloridine, cephalexin, cefazolin, and cephacetrile but only minimally influences the t1/2 of cephalothin and cefamandole. Peritoneal dialysis reduces the t1/2 of cephalothin and cephaloridine but only minimally influences the t1/2 of cefazolin and cefamandole.


Immunofluorescent patterns in the idiopathic interstitial pneumonias.

Immunologic mechanisms have been implicated in the pathogenesis of the IIP's. Thirty five patients with IIP were divided into three groups: group I, seven subjects with DIP; group II, thirteen with UIP; and group III, fifteen with mural fibrosis. Lung biopsies were exposed to fluorescein-conjugated antisera to IgG, IgA, IgE, IgM, C3, fibrinogen, and albumin. In group I, all patients demonstrated IgG and C3 in a granular pattern lining alveolar walls. Similarly, in group II, all had IgG and 10 had C3 within alveolar walls. In contrast, only two group III subjects demonstrated IgG and none C3. Only one of 19 control subjects hav alveolar wall immunoglobulin and C3 deposition. The incidence of positive rheumatoid and antinuclear factors was similar in all three study groups. These data suggest that immune complex deposition may play a role in the pathogenesis of the cellular interstitial pneumonias and that once mural fibrosis supervenes, these complexes are no longer present.


Neonatal respiratory distress. A simple cradle for micrognathia and glossoptosis.

Infants born with micrognathia and glossoptosis may present as a grave emergency, with recurrent bouts of respiratory distress and cyanosis. The management of this condition is discussed, with particular reference to conservative measures. A new cradle has been designed to facilitate the management of these children during the neo-natal period.


Description of obesity in the PBB/Ld mouse.

A new strain of obese mouse, the PBB/Ld, has been studied in terms of fat pad cellularity, serum insulin and blood glucose levels, and response to gold thioglucose injections. Age-matched C57B1/6J mice were used as controls. Adipocyte size and number in the major fat depots were determined at various ages from weanling to maturity in the PBB/Ld and C57B1/6J strains. Results indicated that obesity in the PBB/Ld was due to hypertrophy of adipocytes in retroperitoneal and subcutaneous fat depots and to hypertrophy and hyperplasia in the epididymal fat pad. PBB/Ld mice also developed hyperinsulinemia and hyperglycemia and these findings have been discussed in terms of the developmental changes in fat pad cellularity. The injection of gold thioglucose led to increased food intake in both PBB/Ld and C57B1/6J mice. Hyperphagia was also present in the PBB/LD control group, but increased efficiency of converting calories to body weight was not observed in this group when compared to control C57B1/6J mice. The characteristics of obesity seen in the PBB/Ld mouse are discussed and comparisons are made to similar studies in other rodent models of obesity.


Bacille Calmette-Guérin infection in the mouse. Regulation of macrophage plasminogen activator by T lymphocytes and specific antigen.

High levels of plasminogen activator (PA) were induced in mouse peritoneal macrophages by infection with BCG, 2-6 X 10(7) viable organisms intravenously, followed 3-4 wk later by intraperitoneal challenge with purified protein derivative (PPD) 2 days before harvest. Macrophages obtained from infected animal without boosting showed little fibrinolytic activity, but challenge of Bacille-Calmette-Guèrin (BCG)-primed peritoneal cells with PPD in culture also enhanced macrophage PA 4- to 10-fold. Stimulation of macrophage PA by PPD depended on specifically sensitized thymus-derived (T) lymphocytes because it was abolished by pretreatment of BCG-primed peritoneal cells with anti-thy 1.2 antiserum and complement. A direct assay was developed in which nylon wool separated sensitized lymphocytes and PPD induced PA in macrophages from uninfected animals under defined conditions on 125I-fibrin. Enhanced macrophage fibrinolysis was proportional to concentration of PPD and the number of sensitized lymphocytes transferred. An indirect two-stage assay was also used to show that BCG-sensitized peritoneal cells released a soluble inducer of macrophage PA into the culture medium, after challenge with PPD. Induction of macrophage PA by PPD challenge in vitro made it possible to study the generation and activity of sensitized peritoneal lymphocytes at different stages of infection. Our results show that nonadherent peritoneal cells of BCG-infected mice provide a rich source of specifically sensitized lymphocytes and that macrophage activation is limited by continued availability of antigen, as well as sensitized lymphocytes. Induction of macrophage PA provides a sensitive, versatile, and rapid in vitro assay to study the role of lymphocytes and specific antigen in macrophage activation by BCG.


Hematopoietic stem cell regulatory volumes as revealed in studies of the bgj/bgj:W/WV chimera.

The kinetics of bone marrow replacement was studied in W/WV mice implanted with gbj/bgj (beige) stem cells, with the characteristic beige neutrophil marker as a criterion of the takeover of host marrow by donor marrow. A hyperbolic pattern of W/WV marrow replacement conforming to a log dose-response was observed in experiments encompassing a 50-fold range of bgj/bgj inoculum doses and a 2-yr period of observation. The dose-response relationships were consistent with random seeding of stem cells in the host marrow coupled with a decreasing efficiency of secondary colonization by local migration. Application of single-hit Poisson sampling statistics to the dose-response data led to the hypothesis that mouse bone marrow is compartmentalized into essentially self-contained stem cell regulatory volumes or domains. We estimate that W/WV marrow contains about 2,600 stem cells regulatory units with an average volume of about 10(8) micron3, a dimension consistent with the presumptive role of short-range cell-cell interactions in the regulation of pluripotent stem cells. Our analysis of the dose-response data is also indicative of the discontinuous and limited nature of local stem cell migration in a cellular marrow, a consideration that may be of practical as well as theoretical interest.


Differential effects of polyadenylic: polyuridylic acid and lipopolysaccharide on the generation of cytotoxic T lymphocytes.

In a mixed leukocyte culture (MLC) reaction of allogenic mouse spleen cells differing for H-2K or H-2D, only a weak cytotoxic response is generated. This cytotoxic response is augmented significantly if bacterial lipopolysaccharide (LPS), 5 microgram/ml, or polyadenylic acid (poly A):polyuridylic acid (poly U), 20 microgram/ml, is present in the culture. The cytotoxic cells generated in the presence of these two agents are specific for sensitizing H-2K or H-2D antigen. Two lines of evidence suggest that these two agents exert their effect at different steps in the development of cytotoxic lymphocytes: (a) the effect of poly A:U depends on the presence of adherent cells, whereas the effect of LPS is independent of the presence of adherent cells and (b) LPS promotes the development of cytotoxic cells when ultraviolet light-treated stimulating cells are used in the MLC whereas poly A:U does not.


Primary in vitro cell-mediated lympholysis reaction of NZB mice against unmodified targets syngeneic at the major histocompatibility complex.

T-cell cytotoxicity of NZV mice was tested after in vitro sensitization against a group of H-2 identical strains (BALB/c, B10.D2, DBA/2, HW19). A highly significant and unexpected unidirectional cell-mediated lympholysis (CML) reaction by the sensitized NZB effector cells on these targets was found. After sensitization in vitro with stimulator cells of one H-2d strain, NZB effector cells (H-2d) lysed all other H-2d targets and to a lesser degree, some non-H-2d targets  (C57BL/10, DBA/1, B10.Q, CBA, B10.S, A.SW). NZB targets were not lysed. Differences in the major histocompatibility region between NZB and other H-2d strains could be excluded as a possible explanation for the observed reaction of NZB (H-2d) against other H-2d strains. These results consequently represent the first description of a primary in vitro CML directed against determinants not coded for in the major histocompatibility complex. The responsible effector cells are demonstrated to be T cells. The CML of NZB against H-2 identiical targets appears best explained by a reaction against minor histocompatibility antigens. This, and the observed cross-reactions, would indicate that the cytotoxic T-cell system in NZB mice is not subjected to restrictions found in all normal mouse strains tested until now under similar conditions. It is suggested that this hyperreactivity is related to the autoimmune responsiveness of the NZB strain.


Biochemical genetics of the alpha-keto acid dehydrogenase complexes of Escherichia coli K12: genetic characterization and regulatory properties of deletion mutants.

Twenty-eight spontaneous auxotrophic aroP mutants with deletions in the azi--nadC--aroP--aceE--aceF--lpd region of the Escherichia coli K12 chromosome were characterized genetically with respect to various azi, nadC, ace and lpd markers by P1-mediated transduction. One mutant (Kdelta18; aroP--lpddelta) had a deletion which extended through the aceE and aceF genes to end within the lpd gene. The polarity of the ace operon (aceE to aceF) was confirmed. It was concluded that 10 out of 15 deletions generating a strict requirement for acetate terminated in the aceE gene. Of the ten, three mutants (Kdelta22, Cdelta41 and Cdelta41) synthesized detectable dihydrolipoamide acetyltransferase (the aceF gene product) and seven were assumed to possess deletions generating polar effects on aceF gene expression. Five deletions appeared to extend into the aceF gene. A further five deletions, which limited the expression of the ace operon without generating an Ace- phenotype or a complete Ace- phenotype, ended closest to the aroP-proximal aceE markers. The opposite ends of all these deletions appeared to terminate before (10), within (2) or extend beyond (9) the nadC gene. There was no obvious correlation between the deletion end-points and the corresponding lipoamide dehydrogenase activities, which ranged from 30 to 95% of parental levels in different deletion strains. The remaining seven deletions simply extended between the aroP and nadC genes (nad--aroPdelta) without affecting expression of the ace operon. Regulation of the synthesis of the pyruvate and alpha-ketoglutarate dehydrogenase complexes was investigated in some of the parental and deletion strains under different physiological conditions including thiamin-deprivation. The results indicate that the syntheses of the two dehydrogenase complexes are independently regulated. Expression of the lpd gene appears to be coupled to complex synthesis but can be dissociated under some conditions. Mechanisms for regulating lpd gene expression are discussed and an autogenous mechanism involving uncomplexed lipoamide dehydrogenase functioning as a negatively acting repressor at the operator site of an independent lpd gene is proposed as the simplest mechanism which is consistent with all available information.


Epilepsy, hysteria, and "possession". A historical essay.

A historical essay is presented relating concepts of epilepsy, hysteria, and "possession." The designation "hysteroepilepsy" is placed into the context of combined phenomena in individual subject instances. An association of psychotic states resembling a schizoprenic disorder is indicated as occurring in certain epileptic patients, especially some complex partial seizures (i.e., temporal lobe-psychomotor type). Phenomena of possession may appear within any of these entities. Differential diagnosis now is aided greatly by ulilization of monitoring with combined split screen television viewing and recording of the patient's behavior and the concomitant electroencephalogram. Treatment is directed both medically and toward alleviation of contributing and precipitating psychological and sociological factors.


Dirt-anality and attitudes toward Negroes. A test of Kubie's hypothesis.

Lawrence Kubie's hypothesis that dirt-anal parameters predict attitudes toward Negroes was evaluated. Four samples of males and four samples of females were studied. Multiple indices were used to tap attitudes toward Negroes; and a variety of measures were employed to tap the dirt-anality dimension. The results were significantly supportive of Kubie's formulation.


Ultrastructural localization study of two Wolfgram proteins in rat brain tissue.

The ultrastructural immunohistochemical localization of Wolfgram proteins W1 and W2 is described in young rat brain tissue. The labelling by the antiserum to W1 is restricted to oligodendroglial cells and myelin sheaths. The plasma membrane of the cells as well as the polysomes are positively stained whereas the mitochondria and the nuclei are always free of labelling. Glial cell processes with definite organelles, which are involved in the myelination of neighbouring axons, are also positive to the antiserum. In the myelin sheaths, the positive staining occurs predominantly at the dense period line of the innermost and outermost lamellae. The present results add further evidence for a specific local synthesis of these Wolfgram proteins in oligodendroglial cells during myelination.


Immunocytochemical method to identify basic protein in myelin-forming oligodendrocytes of newborn rat C.N.S.

An immunocytochemical method for detecting myelin basic protein in oligodendrocytes and myelin of newborn rat C.N.S. is described. C.N.S. tissue is perfused and fixes in HgCl2--formaldehyde and 20 micron Vibratome sections are treated with antibodies to myelin basic protein using the peroxidase--antiperoxidase method. Oligodendrocytes in the newborn rat are intensely stained by antiserum to basic protein and multiple stained processes extend from the perikaryon to myelin sheaths. With this procedure it is possible to demonstrate the geometric relationships between a single oligodendrocyte and multiple myelin sheaths. Stained oligodendrocytes and myelin are present in newborn cervical spinal cord, medulla oblongata, pons and midbrain. By 25 days of age, staining in oligodendrocytes is less intense than in newborn rats and differences in amount of staining can be detected in areas that are myelinating at different rates. With anticerebroside serum, cerebroside, of newborn and developing rat C.N.S. tissue is localized only in myelin. In the developing P.N.S., myelin basic protein is localized in Schwann cell cytoplasm and myelin sheaths of the trigeminal ganglion. Cerebroside is found only in myelin.


The psychosocial dimensions of oral and maxillofacial surgery: a critical review of the literature.

The validity of the literature on the relationships between psychosocial factors and surgery for facial esthetics is diminished by theoretical bias, changing cultural values, vague measuring criteria, and inadequate experimental designs. Many persons who seek cosmetic surgery have psychiatric diagnoses and may experience postoperative emotional disturbances. Almost all patients report long-term satisfaction postoperatively. Patients who are treated by oral surgeons appear to have fewer emotional problems than those treated by plastic surgeons.


Posterior mandibular segmental alveolar osteotomy.

A technique for mobilizing and repositioning mandibular posterior alveolar segments is described. These segments can be extruded or impacted, tipped bucally or lingually, or can be uprighted. Two case reports illustrate the technique.


Tetanus--review of the literature and report of case.

This case presents many of the difficulties of management that are inherent in a severe case of tetanus. Also it shows a seldom-considered therapeutic modality, that of dental extraction, gingival debridement, and gingivectomy with a confirmed case of tetanus without an established portal of entry. It is well known that periodontal and periapical locations can easily give rise to an anaerobic focus. In this case, anaerobic culture of C tetani was unsuccessful, possibly because of the inherent difficulty of anaerobic transfer from an oral locus and the extreme fastidiousness of the organism. As a rule, all patients who recover from tetanus do so completely, without any residual deficit. This was true in the case discussed here. Conclusions that can be drawn from this case are the following: a diagnosis of tetanus should be considered in any case of unexplained trismus; aggressive treatment of the immunologic deficit by antitoxin should be instituted along with a course of active immunization; and aggressive symptomatic treatment should be started. The bases of the treatment of this case were management of airways, sedation, treatment of autonomic crisis, and eradication of the causative agent. The result was that a severely ill patient reponded to treatment and completely recovered from a devastating disease.


Utilization of pressure-volume curves in the pediatric patient.

Seven patients treated with continuous mechanical ventialtion were monitored with static and dynamic pressure-volume curves. Three patients developed no pulmonary complications, and mechanical ventilation was discontinued within 96 hr. In four patients, pressure-volume curves were used as a diagnostic aid in the detection of the physiologic defect resulting from bronchoconstriction, atelectasis, loculated pleural fluid, pulmonary edema, and mucous plugging. These measurements were also utilized to evaluate the effectivess of therapeutic modalities such as treatment of bronchoconstriction with bronchodilators, mucous plugging with adequate suctioning, and drainage of loculated pleural effusion. Pressure-v-lume measurements are simple, noninvasive, and require the smae equipment used in continuous mechanical ventilation. Pressure-volume monitoring of pediatric patients with curves warrants further investigation to evaluate its value.


Studies on kallikrein in the duct systems of the salivary glands of the cat.

By correlating immunofluorescence light microscopy with electron microscope studies and with kallikrein concentrations under various conditions, we have made the following observations and conclusions about kallikrein in the submandibular and other salivary glands.1. In the submandibular gland, specific immunofluorescence to kallikrein was observed in the luminal region of the striated ducts particularly, but also in the outer epithelial cells of the stratified epithelial collecting ducts. Sympathetic nerve stimulation resulted in a reduction in intensity of specific fluorescence and in its increased localization towards the lumen. The nearly complete elimination of kallikrein from the gland by duct obstruction for four days resulted in complete disappearance of specific fluorescence in the gland. Prolonged parasympathetic nerve stimulation at frequencies which did not reduce the kallikrein concentration of the gland failed to alter the specific immunofluorescence despite copious secretion of saliva. Our results failed to reveal evidence of secretion of kallikrein either into or towards the interstitium of the gland. The luminal layer of stratified epithelial cells in the collecting ducts contained small secretory granules closely resembling those in the striated ducts. Our results are not conclusive, but do suggest that kallikrein is located in these granules whence it is secreted into the lumen of the duct.2. The parotid gland was found to contain much lower concentrations of kallikrein than the submandibular gland. This finding was associated with the presence of far fewer striated ducts in the parotid gland. Otherwise, specific fluorescence and the response to sympathetic nerve stimulation was like that of the submandibular gland. Small secretory granules in the striated and collecting ducts resembled those of the submandibular gland.3. The sublingual gland, like the parotid, had a low concentration of kallikrein and very few striated ducts. These ducts were unevenly distributed and were concentrated in only a few lobules of the gland. Specific immunofluorescence was seen only in sections containing striated ducts.4. The possible physiological role of kallikrein in the salivary glands is discussed.


Structure-activity relationships of eighteen somatostatin analogues on gastric secretion.

1. The effect of somatostatin and eighteen somatostatin analogues on pentagastrin-stimulated gastric acid and pepsin secretion was investigated in the conscious vagotomized cat prepared with chronic gastric fistulae. The majority of the analogues are peptides where D-amino acids are incorporated into the molecule instead of the natural L-isomers. 2. The ID50 for cyclic-somatostatin inhibition of near-maximal gastric acid secretion stimulated by pentagastrin 8 microgram kg-1 hr-1 was found to be 1.29 +/- 0.13 n-mole kg-1 hr-1. Pentagastrin-stimulated pepsin secretion had a lower threshold to somatostatin inhibition than did acid secretion. 3. D-Phe6, D-Phe7, D-Thr10, D-Thr12 and D-Phe6-D-Trp8 analogues all show low biological activity against the secretion of gastric acid and pepsin, growth hormone, insulin and glucagon. None of these analogues are antagonists of the cyclic-somatostatin inhibition of gastric secretion, suggesting that they have low affinity for this somatostatin receptor. 4. The analogues under investigation show parallel changes in activity against gastric and growth hormone secretion, suggesting a similarity between the gastric and growth hormone receptors for somatostatin. 5. D-Cys14 analogues are equipotent with or have a greater potency than cyclic-simatostatin in inhibiting the secretion of gastric acid, growth hormone and glucagon but show low insulin inhibiting activity.


Arrangement of anatomic-type artificial teeth into balanced occlusion.

The mechanical factors which are involved in the arrangement of anatomic type artificial teeth into balanced occlusion have been described. Many of the mechanical problems encountered during artificial tooth arrangement have been listed, together with suggestions for their solution. Hanau's Laws of Articulation have been utilized in a practical manner in formulating some of the solutions to tooth arrangement problems when establishing balanced occlusion. Trial dentures which present ideal occlusions in the articulator may not always do so when examined in the patient's mouth. Possible reasons for these differences are listed, and methods for making the necessary corrections are described.


The kinetics of anterior tooth display.

A survey has been presented that correlates measurements of upper lip type, sex, race, and age of dentulous patients with the amount of exposure of the maxillary and mandibular anterior teeth with the lips gently parted and in the resting position. Perhaps the most interesting finding was the gradual reduction in the amount of maxillary central incisor exposure with an increase in age, accompanied by a gradual increase in the mandibular tooth exposure. The importance of the amount of mandibular teeth seen in complete dentures has not been sufficiently emphasized in previous literature.


Inconspicuous retention for removable partial dentures.

Many dentists and patients believe that removable partial dentures have an unesthetic appearance when they replace anterior teeth because of the visible clasps. Two methods of obtaining inconspicuous retention have been described.


A comparison of pin-retained amalgam and composite resin cores.

The 24 hour tensile loads required to separate the core material and dentin produced the following results: 1. No statistical difference was found between the tensile strength of retained amalgam and composite resin core materials. 2. Generally, no statistical difference was found between the retentive ability of three and four pins. 3. No statistical difference was found between the retentive ability of regular pins and minim pins if three or more pins were employed. 4. As the number of regular pins was increased beyond three, and when four minim pins were used, there was a very significant increase in dentin fracture.


Scanning electron microscope investigation of in vivo performance of eight composite resins.

Retention. At 6 months, five restorations were lost due to retention failure. The remaining restorations were retained to 24 months and/or 30 months. The study of one material was discontinued because of numerous retention failures. Marginal wear. Facial wear along the margin in the center portion of the restoration was appreciably greater than lingual, proximal, and incisal wear. In general, most composite resins began to display evidence of wear during the first 6 months of service. Over the study period, three restorations were lost because of excessive marginal wear. Surface texture. Most materials were clinically acceptable with regard to surface texture over the course of the study. Extended studies of greater durations are indicated.


Fabrication of a single anterior intermediate restoration.

A technique of modifying a polycarbonate shell crown with an acrylic resin reline has been presented. The advantage in using this procedure is that it results in a satisfactory treatment restoration that is functionally and esthetically acceptable to the patient.


Metal posterior teeth for the chronic bruxing patient.

This article outlines a technique for the fabrication of metal posterior tooth blocks which have tooth-colored facings of acrylic resin or dental porcelain. The technique is applicable for all removable dental restorations but has special applicability for the chronic bruxing patient who has a natural dentition opposing the removable partial denture.


The relationship of women's sex-role identity to self-esteem and ego development.

The present study examined the relationship of women's sex-role identity to self-esteem and ego development. One hundred fifty-three female undergraduate students at the University of Maryland were the volunteer Ss for this study. During class time these persons completed the Index of Adjustment and Values, used to measure self-esteem, and the Sentence Completion Test, ussed to measure ego development. From this pool of Ss four sex-role identity groups were determined: androgynous women, feminine women, masculine women, and undifferentiated women. Twenty-five from each group were randomly selected to comprise a final sample of 100 women. Results of the self-esteem analyses supported previous findings that androgynous women possess a higher degree of self-esteem than stereotypically feminine women and undifferentiated women, but revealed no significant difference in self-esteem between androgynous and masculine women. In contrast, the level of ego development of androgynous women was not significantly different from that of feminine and undifferentiated women but was significantly higher than that of masculine women. Overall results support the theory that a combination of masculine and feminine characteristics is beneficial for women in terms of both self-esteem and ego development.


[Therapeutic arterial embolization in hemobilia. Apropos of 3 cases].

Three patients with bleeding into the bile ducts were explored by angiography. Two of them (severe post-operative bleeding and following needle-biopsy of the liver) were completely cured by highly selective arterial embolus production. No definite diagnosis was established in the third case (liver angioma with large artero-portal fistula). It can be seen from these observations, that angiography is essential in patients with bleeding into the bile ducts. The reason is not to establish the diagnosis but to find the precise location of the lesion and, under certain conditions, to apply radical treatment by highly selective arterial embolus production.


Effects of rubidium chloride on the course of manic-depressive illness.

Clinical studies of the effects of rubidium ions on the course of manic-depressive illness are reported. It seems that rubidium tends to increase the length of manic phases and possibly reduces the extremes of mood. Rubidium did not seem to produce any severe side effects in the dose administered, but it has a long biological half-life and caution is still required. Some details of the CSF, RBC, saliva and plasma and urine kinetics are also reported.


Resolution of antimalarial agents via complex formation with alpha-(2,4,5,7-tetranitro-9-fluorenylideneaminooxy) propionic acid.

The resolution of several antimalarial agents via pi-complex formation with alpha-(2,4,5,7-tetranitro-9-fluorenylideneaminooxy) propionic acid (TAPA) is reported. Since this represents the first use of this agent for the resolution of amines, some details of the separations are presented. The method proved successful for resolving weakly alkaline amines that did not form stable salts with common resolving acids, highly insoluble amines that did not form soluble salts with usual resolving acids, and amines that did not form crystalline salts with commonly available resolving acids. The optical isomers of several antimalarial agents were evaluated against Plasmodium berghei in the mouse. None of the optically active forms showed any significant differences. The curative activity of (+)- and (-)-primaquine against Plasmodium cynomolgi in the rhesus monkey was essentially identical; however, significant differences in toxicity were noted.


Folate antagonists. 10. Synthesis and antimalarial effects of 6-[[(aryl and aralkyl)amino]methyl]-2,4-pteridinediamines and -pteridinediamine 8-oxides.

Various 6-[[(aryl and aralkyl)amino]methyl]-2,4-pteridinediamines and their 8-oxides have been synthesized for antimalarial evaluation. Condensation of 3-amino-6-(bromomethyl)-2-pyrazinecarbonitrile 4-oxide (V) with the appropriately substituted amine afforded a series of 3-amino-6-[[(aryl and aralkyl)amino]methyl]-2-pyrazinecarbonitrile 4-oxides VI. Deoxygenation gave the corresponding pyrazines VII. Cyclization of VI and VII with guanidine then produced the desired 6-(aminomethyl)-2,4-pteridinediamine N-oxides VIII and teridinediamines IX, respectively. Formylation of 6-[[(3,4-dichlorophenyl)amino]methyl]-2,4-pteridinediamine gave N-[(2,4-diamino-6-pteridinyl)-methyl]-N-(3,4-dichlorophenyl)formamide. The N-oxides VIII did not exhibit significant activity against Plasmodium berghei infections in mice. Activity among the 2,4-pteridinediamines IX was generally poor with the exception of the 3,4,5-trimethoxyphenyl and 1-naphthalenyl analogues which showed strong suppressive activity at doses ranging from 80 to 640 mg/kg. Furthermore, several of the 2,4-pteridinediamines exhibited potent prophylactic activity against Plasmodium gallinaceum infections in the chick and also showed strong antibacterial action against Streptococcus faecalis and Staphylococcus aureus.


Folate antagonists. 11. Synthesis and antimalarial effects of 6-[(aryloxy- and arylthio-)methyl]-2,4-pteridinediamines and -pteridinediamine 8-oxides.

Condensation of 3-amino-6-(bromomethyl)-2-pyrazinecarbonitrile 4-oxide with 4-chlorophenol gave 3-amino-6-[(4-chlorophenoxy)methyl]-2-pyrazinecarbonitrile 4-oxide (1), which was deoxygenated to obtain the de-N-oxide 4. Cyclization of 4 and 1 produced 6-[(4-chlorophenoxy)methyl]-2,4-pteridinediamine and the 8-oxide, respectively. 6-[(Arylthio)methyl]-2,4-pteridinediamines and their 8-oxides were produced analogously. Controlled oxidation of the former gave the anticipated sulfoxide 12 and sulfone 13. None of these compounds showed significant activity when tested against lethal Plasmodium berghei infections in mice or a select list of bacteria in vitro.


Structure and function of frozen cells: freezing patterns and post-thaw survival.

Freezing patterns and post-thaw survival of cells varies with different cooling rates. The optimal cooling rates, indicating the highest percentage survival, were different in yeast and red blood cells. A difference of freezing patterns was also noticed in preparations frozen above and below the optimal cooling rate for each cell, namely, cell shrinkage at lower rates and intracellular ice formation at higher rates which showed similar trends in both the cells, even though there was some shifting of the optimum. Ultra-rapid freezing and addition of cryoprotectants are useful ways to minimize ice crystal formation and to cause such ice formations to approach the vitreous state. Ice crystals are hardly detectable in yeast cells as well as in erythrocytes, when these cells are frozen ultra-rapidly in the presence of cryoprotective agents in moderate concentration.


Two rapid and simple methods used for the removal of resins from 1.0 micron thick epoxy sections.

Ultrathin epoxy sections are commonly used in electron microscopy. More useful information can often be obtained by examining thicker 0.5--3.0 micron) resin sections under the light microscope. However, there are some histological stains that are unable to penetrate these resins. This short note describes the easy and rapid preparation of two resin-removal solutions which allow subsequent staining of 1.0 micron thick sections without obvious tissue damage and with excellent results using standard histological dyes.


Blocking plasma factors of migration inhibition factor production in patients with cervical carcinoma. A possible role in vivo.

The blocking activity was studied of plasma from patients with cervical carcinoma in a model of migration inhibition factor (MIF) production by normal human leukocytes. At the same time, the patients' in vivo (intradermal reactivity) and in vitro (MIF assay) responses were studied with two common antigens (streptokinase-streptodornase and purified protein derivative). The plasma of 15 of 27 patients with carcinoma was capable of blocking the MIF test of human normal leukocytes. Twelve of 15 patients with blocking factor in their plasma had no or only a slight in vivo response, although they had a good in vitor response. Two patients had blocking factor without any in vivo or in vitro response. Six patients who had in vitro responses and slight or no in vivo responses had no plasma blocking activity. Three patients had neither in vivo nor in vitro responses and lacked blocking activity in their plasma.


Tissue localization of zinc glycinate marker and carcinoembryonic antigen by immunofluorescence. I. Preparation of antisera against the zinc glycinate marker.

Antisera against the zinc glycinate marker (ZGM) were produced in New Zealand White rabbits with induced tolerance to normal tissue components and CEA, and in mature, previously uninoculated rabbits for use in immunofluorescent histologic localization of ZGM in colon cancers and other tissues. Analysis of the antisera by immunodiffusion and counterimmunoelectrophoretic techniques showed them to be specific for ZGM when tested with ZGM, carcinoembryonic antigen, normal tissue extracts, or cell elements of normal blood.


Tissue localization of zinc glycinate marker and carcinoembryonic antigen by immunofluorescence. II. Immunofluorescence microscopy.

Preliminary indirect immunofluorescence studies on the zinc glycinate marker (ZGM) were compared with carcinoembryonic antigen (CEA) immunofluorescence, ZGM, detected in 26 of 29 human colon adenocarcinomas, was associated with the epithelial component of the malignant glands. Fluorescence was generally less strong and more granular for ZGM than for CEA and was found in intraglandular spaces, luminal border areas, and cytoplasm. ZGM concentration and tissue localization appeared to be related to tumor differentiation. ZGM was also detected in benign colon mucosae (adjacent to and distant from the carcinomas) from patients with colon carcinoma, but differed from CEA in that it was present in the deep crypt portion only. Gastric, pancreatic, esophageal, and anal adenocarcinomas, as well as benign gastric pyloric and small bowel mucosae had detectable ZGM. CEA, but not ZGM, was observed in 20 nongastrointestinal carcinomas to date. Studies are under way to determine whether ZGM is a marker associated with colon and gastrointestinal adenocarcinoma specifically or undifferentiated crypt cells of the colon and digestive tract in general.


Mutagenicity of nitrosourea compounds for Salmonella typhimurium.

The nitrosourea derivatives 1,3-bis(2-chloroethyl)-1-nitrosourea (NSC-409962), 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea (CCNU; NSC-79037), 1-(2-chloroethyl)-3-(4-methylcyclohexyl)-1-nitrosourea (Me-CCNU; NSC-95441), and 1-methyl-1-nitrosourea (MNU; NSC-23909) were screened for mutagenic potential with Salmonella typhimurium strains G46 and TA1530 in vitro and in vivo. Alu were also mutagenic in the host-mediated assay. Four additional chemotherapeutic nitrosoureas were tested in vitro, three of which were mutagenic. The lack of activity of the fourth agent was probably a reflection of its stability in solution rather than a true indication of mutagenic potential. All eight agents were tested in a repair test with S. typhimurium strains TA1978 and TA1538. Results of this test were negative, reflecting the insensitivity of these strains to ethylating and methylating agents. The insensitivity of the host-mediated assay is also discussed.


Facial reconstruction using local skin flaps.

The use of local skin flaps for reconstructing small to moderate size surface defects of the facial area is presented. The indications for the use of skin flaps, the local and general factors involved in selecting the best flaps for reconstruction, and proper planning of the flaps are discussed. Basic local skin flaps are illustrated by diagrams and clinical cases are presented to demonstrate the use of the different type flaps.


Angiography in the diagnosis of renal allograft dysfunction.

We reviewed retrospectively 75 renal transplant arteriograms done during a 7-year period. Acute rejection and vasomotor nephropathy were not differentiated. Generalized cortical ischemia was diagnosed correctly in 23 of 30 cases but there were 7 falsely negative results. Renal artery stenosis was found in 7 of 17 cases in which the main indication for arteriography was hypertension. We conclude that the major role of transplant arteriography is in the diagnosis of larger vessel disease.


Experience with the Horton-Devine dermal graft in the treatment of Peyronie's disease.

We treated 15 patients with Peyronie's disease surgically with the dermal graft technique described by Horton and Devine. Details of the preoperative management and surgical technique are provided. Return of normal sexual function without residual chordee or pain was achieved in more than 75 per cent of the patients, suggesting that this is the procedure of choice in the management of severe Peyronie's disease.


Symptomless acute renal transplant rejections. Occurrence six months or more after transplantation.

A total of 22 cases of acute rejection episodes occurring six months to five years after renal transplantation were retrospectively reviewed for the presence of fever, graft tenderness, increase in blood pressure, declining urine output, falling urine sodium level, change in WBC count, and patient's response to antirejection therapy. A total of 13 episodes were not associated with any of these symptoms or signs of an acute rejection episode; seven episodes were associated with an increase in blood pressure alone and only two episodes were associated with fever. Nineteen rejection episodes were confirmed by biopsy. In 16 of these late-rejection episodes, the patient had complete or partial response to therapy. Acute rejection episodes can occur months to years after transplantation; patients are typically asymptomatic; many patients do respond to therapy; and biopsy is often helpful in establishing the diagnosis.


Diagnostic value of sinus-tract cultures in chronic osteomyelitis.

Sinus-tract cultures were compared with cultures of operative specimens from 40 patients with chronic osteomyelitis. Thirty-five patients (87.5%) had a single pathogen isolated from their operative specimens. Only 44% of the sinus-tract cultures contained the operative pathogen. Isolation of Staphyloccus aureus from sinus tracts correlated with the presence of S aureus in the operative specimen. However, less than half of the sinus-tract cultures obtained from patients with S aureus osteomyelitis contained this organism. Isolation of bacteria other than S aureus from sinus tracts had a low likelihood of predicting the pathogen isolated from bone. A presumptive diagnosis of S aureus osteomyelitis is justified if S aureus is isolated from an associated sinus tract. A bacteriologic diagnosis of chronic osteomyelitis based on isolation of common pathogens other than S aureus from sinus tracts must be verified by an appropriate operative culture.


Falciparum malaria in seamen.

Three cases of Plasmodium falciparum malaria in seamen, all acquired while working off tropical West Africa, and all in patients coming in for treatment at a New Orleans hospital during one six-week period, are described in the context of the importance of considering recent travel history for arrival at the correct diagnosis and treatment. Two of the three patients whose cases are reported had taken some form of malarial chemoprophylaxis during their voyage.


Persistent nocardemia following renal transplantation. Association with pulmonary nocardiosis.

Pulmonary nocardiosis occurred in a renal homotransplant patient and was diagnosed by persistently positive blood cultures. Infection developed at a time when efforts were being made with moderate dosages of immunosuppressive drugs to prevent rejection of the transplant. The subsequent nocardemia lasted for one week. The remaining kidney function was preserved, and the patient had a very favorable outcome because of early diagnosis and prompt treatment with sulfisoxazole.


Management and microbiology of cutaneous abscesses.

Cutaneous abscesses in the perineal region contain primarily anaerobes commonly found in stool. Non-perineal abscesses contain mixed microflora indigenous to the skin. Bacteroides fragilis, the only anaerobe resistant to penicillin, is found predominantly in the perineal area. Staphylococcus aureus, although the most common aerobe found, is seen less frequently than expected and is almost always resistant to penicillin. Escherichia coli and Neisseria gonorrhoeae are rarely found. Incision and drainage is the primary mode of treatment. Fever is rare, though tachycardia is common in patients with normal host defenses. Patients with altered host defenses may require initial culture and Gram-stained smear to determine appropriate antibiotic therapy. The Gram-stained smear reliably indicates sterile and mixed abscesses, as well as those containing pure S aureus. Incision and drainage alone is the only therapy required in the usual patient. Since in these patients antibiotics are not required, initial culture and Gram-stained smear are unnecessary.


Study of chemical sympathectomy in endotoxin-induced lethality and fibrin depostion.

Shock and the generalized Shwartzman reaction are well known features of endotoxin which have been shown to involve the sympathetic nervous system. The mechanism of sympathetic nervous system involvement with endotoxin injection was studied in rabbits chemically sympathectomized with 6-hydroxydopamine. Endotoxin, in doses producing a spectrum of morbidity and mortality in normal rabbits, was administered i.v. to chemically sympathectomized, normal, and unilateral renal surgically sympathectomized animals. Chemical sympathectomy produced a significant depletion of tissue norepinephrine which, in endotoxin recipient animals, was associated with a significantly lower mortality rate and greatly decreased fibrin deposition in the lungs and kidneys, despite intravascular coagulation. Unilateral renal sympathectomy afforded protection to the ipsilateral kidney, but data on mortality and systemic fibrin deposition were similar to those reported for normal rabbits given endotoxin. Six-hydroxydopamine prevents significant tissue injury secondary to endotoxin in this experimental model. In addition, the data provide direct evidence that an intact reactive sympathetic nervous system is essential for development of lethal toxicity and generalized Shwartzman reaction due to endotoxin.


Microangiographic evaluation of the effects of heparin on progressive Masugi nephritis.

Unilateral progressive Masugi nephritis was produced in rabbits and studied by microangiography as well as light and immunofluorescence microscopy. The following five groups were studied: Group 1. Heparin was started simultaneously with nephrotoxic serum (NTS) and was given for one week. The animals were then sacrificed along with the untreated controls. Group 2. This was the same protocol as in group 1 but with three weeks' heparin. Group 3. Heparin was started one day after NTS was given for three weeks. Group 4. Heparin was started one to two weeks after NTS and was given for three weeks. Group 5: late effect group. Heparin was started simultaneously with NTS, was given for four weeks, and the animals were then sacrificed 10 to 13 weeks later. Heparin dose was 5,000 U, s.c., per day in all treated groups. The number of glomeruli seen per unit of cortex by microangiography was significantly increased in the first through the third groups, as compared to the controls. Group 1 did not show this increase but there was some decrease of immunofluorescent fibrinogen. The late effect group (group 5) showed no modification by the treatment, suggesting that an initial improvement may have been negated by persistent immunologic insults after heparin withdrawal.