Rosetting and other reactions of the Reed-Sternberg cell. Clustering of lymphocytes around Reed-Sternberg cells was noticed in single cell suspensions made from viable Hodgkin's lymphoid tissue. Cytocentrifugation of the suspension showed that clustering also occurred around a smaller cell type, thought to be the precursor of the classical Reed-Sternberg cell. Time-lapse cine films taken of the clustering showed unceasing activity on the part of the lymphocytes migrating over the surface of the central cell. Reed-Sternberg cells were reacted with anti-monocyte serum using indirect fluorescence techniques. In its mature form at least, the Reed-Sternberg cell showed no activity with the antiserum. No immunoglobulin was detected in the Reed-Sternberg cell using fluorescence techniques, but a few Reed-Sternberg cells showed diffuse cytoplasmic staining using the peroxidase-labelled antibody technique. Membrane receptor tests showed the lymphocytes surrounding the Reed-Sternberg cell to be T-cells. After proteolytic enzyme treatment to free lymphocytes from the surface, the Reed-Sternberg cell bound IgG-coated red blood cells indicating a probable Fc receptor. Cytochemistry demonstrated weak non-specific esterase activity in a small minority of Reed-Sternberg cells, and absence of acid phosphatase, alkaline phosphatase and peroxidase. A subpopulation of lymphocytes with distinctive segmentation of the nucleus was noted. These were often to be seen participating in lymphocyte rosettes around the Reed-Sternberg cell.


The effect of gestational age on intralipid tolerance in newborn infants. Tolerance to intravenously administered Intralipid was studied in 18 newborn infants aged 4 to 8 days and gestation 27 to 40 weeks. Clearance of a bolus injection of Intralipid from plasma was an exponential function following first order kinetics. Less mature infants had a clearance rate less than half that seen in more mature infants. Free fatty acids rose more rapidly and were maintained above fasting levels throughout the study in the immature group, probably because of impaired utilization of free fatty acid. The slower clearance of Intralipid in immature infants may be related to lipoprotein lipase deficiency secondary to their limited adipose tissue mass.


Increased hypoxemia in neonates secondary to the use of continuous positive airway pressure. Five neonates are presented who, while receiving continuous transpulmonary pressure, developed increased systemic arterial hypoxemia, which was relieved when this therapeutic modality was decreased or discontinued. Three of the five had chest radiographs consistent with hyaline membrane disease: the other two had atypical radiographs. None had evidence of air leaks. Levels of positive airway pressure utilized when the hypoxemia was noted ranged from 5 to 15 cm H2O. Calculated shunt fractions (Qs/Qt) improved from a mean of 73% to 37% when the continuous positive airway pressure was lowered. The site of shunting in these infants could have been in the lung or through anatomic extrapulmonary shunts. Improvement in total cardiac output may also have contributed to the improved oxygenation. This paradoxical effect of continuous transpulmonary pressure appears to be mediated through changes in the pulmonary vascular resistance brought about by the continuous transpulmonary pressure.


Heat-stable enterotoxigenic Escherichia coli and necrotizing enterocolitis: lack of an association. During an outbreak of diarrhea in a special care nursery caused by heat-stable enterotoxigenic Escherichia coli (serotype 078:H11:K80), nine (4.3%) of the 205 infants in the nursery developed necrotizing enterocolitis. Cases of necrotizing enterocolitis were not significantly more common in infants colonized or infected with these organisms; heat-stable enterotoxigenic E. coli was isolated from 5(56%) of nine cases of necrotizing ecterocolitis and from 27(38%) of the 71 infants without necrotizing enterocolitis who were also cultured. Our findings suggest that caution should be taken in implicating enterotoxigenic E. coli as a cause of necrotizing enterocolitis.


Spectrophotometry as a tool for assaying endotoxins. A spectrophotometric method is reported for assaying endotoxins over a wide concentration range, including low parts per billion levels. Five solution of endotoxins. Escherichia coli 0127:B8, Escherichia coli 055:B5, Salmonella abortus-equi, Salmonella enteritidis, and Shigella flexneri, were examined. Each exhibited an absorption maximum at 259 nm, but the absorptivities differed in each case. Thus, rigorous quantification requires that the identity of the endotoxin be known. The uv absorbance and the statistical data for the standard curves for three groups of standard solutions are presented. The absorbance for each group was linear with concentration within acceptable limits.


Sex differences in drug evaluations.

Sex differences in drug evaluations. Twenty normal, nonobese subjects (10 males and 10 female) were administered a battery of seven psychomotor tests as well as affect checklists and physiological measurements on 6 alternate days. Subjects performed the entire battery predrug and at 45,90, and 135 min postdrug. Fenfluramine, dextroamphetamine, two combined doses of the drugs, and a placebo were given in a double-blind, repeated measures design. Findings revealed significant sex-based differences in initial performance on five of the seven psychomotor tasks and in two physiological measures, with males performing at higher levels than females. Additionally, sex differences in postdrug changes were found in three psychomotor and two physiological measures, with females evidencing greater change scores than males.


Immunoreactive luteinizing hormone releasing factor (LRF) in pituitary stalk plasma from female rats: effects of stimulating diencephalon, hippocampus and amygdala. 1. Previous studies on the effect of preoptic and median eminence stimulation on the immunoreactive LRF content of pituitary stalk blood from pro-oestrous rats have been extended. Stimulation of the suprachiasmatic nuclei and anterior hypothalamic area produced increments in LRF which were 66 and 18% respectively, of that produced by preoptic stimulation, and 38 and 9%, respectively, of that produced by stimulation of the median emience. Stimulation of the amygdala and hippocampus had no effect. 2. The LRF response was not affected significantly when preoptic stimulation was accompanied by stimulation of the hippocampus. 3. In animals subjected to section of the dorsal afferents of the diencephalon, the LRF response to preoptic stimulation was similar to that in intact rats. However, the facilitatory effect of oestrogen on the LRF response to preoptic stimulation was significantly reduced in the roof sectioned compared with intact animals. The post-operative resumption of oestrous cycles was delayed but not abolished by dorsal deafferentation.


The functional significance of sodium channels in pancreatic beta-cell membranes. 1. The existence and functional significance of Na channels in pancreatic beta-cell membranes were investigated by studying the effects of the plant alkaloid veratridine on the temporal release of insulin from perfused isolated rat islets of Langerhans.2. 100 muM veratridine evoked a sustained threefold increase in insulin release which was almost completely inhibited by 3 muM tetrodotoxin (TTX). This action of TTX was rapidly reversible.3. The simultaneous presence of 100 muM propranolol, 100 muM phenoxy-benzamine and 10 muM atropine did not alter the magnitude of the response to 100 muM veratridine, indicating that the action of veratridine on the beta-cells was direct and was not mediated via the release of neurotrans-mitters from nerve endings within the islets.4. (45)Ca uptake by isolated islets in static incubation was increased almost threefold by 100 muM veratridine. This increase was completely inhibited by the simultaneous presence of 3 muM TTX.5. Replacement of Na(o) by choline caused a transient fourfold increase in insulin release which was associated with an increase in the uptake of (45)Ca from the extracellular space of similar magnitude. Subsequent exposure of islets to 100 muM veratridine still evoked some insulin release but this only achieved 32% of that secreted by islets exposed to veratridine in medium of normal [Na](o).6. The addition of 2.5 mM CoCl(2) to the medium caused a 62.5% inhibition of veratridine-mediated insulin release.7. In Ca-free medium supplemented with 1 mM EGTA, 100 muM veratridine evoked insulin release of equal magnitude and of similar temporal relationship to that obtained in the presence of normal [Ca](o).8. A twofold increase in insulin release that occurred in the 15 min period immediately following exposure to 1 mM ouabain was completely independent of [Ca](o). Subsequent ouabain-evoked release became increasingly dependent on [Ca](o).9. Tetrodotoxin (3 muM) inhibited the first phase of insulin release evoked by 16.7 mMd-glucose by 37% and the second phase by 20%.10. Both Na and Ca appear capable of entering through Na channels opened in the beta-cell membrane by veratridine. The increase in [Na](i), resulting from the veratridine mediated increase in P(Na+), causes depolarization of the beta-cell membrane with a consequent opening of voltage-sensitive, Co(2+)-blockable channels for additional Ca entry. An increase in [Na](i) also increases [Ca](i) by altering the equilibria of intracellular Ca-sequestering mechanisms. The small but significant reduction of glucose-mediated insulin release by TTX indicates that glucose has a rather weak action on the Na channel and a more pronounced effect on the voltage-dependent Co(2+)-blockable Ca channel.


Elective manual exploration of the uterus after delivery: a study and review. One hundred patients who had elective manual exploration of the uterus at the time of delivery were compared with a control group of 100 patients with regard to febrile morbidity and blood loss at delivery. Febrile morbidity was quantitated with the Fever Index, and postpartum blood loss was quantitated by subtracting the two-day postpartum hemoglobin from the admission hemoglobin. The patient population encompassed only normal spontaneous vaginal deliveries without complications. The manually explored group had less febrile morbidity and less blood loss; however, this was not statistically significant. Although this study cannot conclude that routine elective manual exploration of the uterus should be done after expulsion of the placenta, it does demonstrate the safety of the procedure.


[Newer veterinary neuropathological conditions in South Africa (author's transl)]. A brief review is given of some of the more recent findings on lesions of diseases affecting the central nervous system of animals. These include local diseases caused by poisonous plants; Helichrysum argyrosphaerum, Solanum kwebense, Matricaria nigellaefolia; infectious diseases e.g. heartwater, cerebral babesiosis and cranial abscesses in goats caused by Corynebacterium pyogenes. Reference is also made to some conditions, originally described in overseas countries and which subsequently have been diagnosed in South Africa. Original findings on lesions seen in mycotoxicoses caused by Fusarium moniliforme and Aspergillus clavatus are included.


[Renal cancer embolization with isobutyl-2-cyanoacrylate. Experimental study and first clinical applications (author's transl)]. Isobutyl-2-cyanoacrylate (IBC) is an acrylic monomer which quickly polymerizes in contact with blood. It has been rarely used as an embolic material and never in embolization of renal cancer. The authors report their experience on the use of this substance as embolic material, based on an experimental study in the rat and on two human cases of renal cancer, where a preoperative transcatheter embolization was performed. IBC revealed as a substance which can produce a very efficient and lasting embolization. No complications were observed.


Inhibition of the activity of the luteinizing hormone-releasing hormone (LH-RH) by analogues with variations at positions 2, 3, and 6 and the carboxyl terminus. In the isolated rat pituitary assay, [Thr2,Leu3]-LH-RH, [Leu2,Ala3,D-Ala6]-LH-RH, and des-Gly10-[Abu2,Ala3,D-Ala6]-LH-RH ethylamide inhibited the LH release due to 0.3 ng/mL of added LH-RH at a 10 microgram/mL dosage. Under these same assay conditions, des-Gly10-[Ile2,Ala3,D-Ala6]-LH-RH ethylamide was about one-tenth as active, and no inhibition was observed by [Leu2,Ser3]-LH-RH or [Leu2,Asn3]-LH-RH at a 100 microgram/mL dosage. The corresponding results from FSH inhibition assays, in vitro, are also reported.


Differences in antischistosomal and mutagenic properties between an isothiocyano- and an isocyanonitrodiphenylamine. While 4-isothiocyano-4'-nitrodiphenylamine has high schistosomicidal activity in vivo and is devoid of mutagenic properties in vitro, the reverse is true for the isocyano analogue of this compound; i.e., replacement of the sulfur by oxygen results in a compound that has no demonstrable antischistosomal effects and exhibits significant mutagenic activity.


One hundred and twenty-seven cultured human tumor cell lines producing tumors in nude mice. One hundred and twenty-seven cultured human tumor cell lines produced tumors after sc inoculation of 1-20 million cells into nude mice. They included 56 carcinoma lines, 14 sarcoma lines, and 57 lines from miscellaneous tumors and were all glucose-6-phosphate dehydrogenase type B. Twenty-nine percent of the lines produced tumors of 1 cm3 size within 1 month and 41% in the second month after inoculation. The histopathology correlated with the human tumor of origin in all cases.


Group B beta-hemolytic streptococci analysis of 242 isolations. Group B beta-hemolytic streptococci (GBS) were isolated from 242 patients at Howard University Hospital during a seven-month period. The source of the majority, 236, was the genital tract of women. In six instances, the organism was cultured from various sites in neonates born of mothers harboring GBS in their birth canals. All of the patients were black. The study indicates that the frequency of colonization of the genital tract by GBS in black females is higher than that reported for white women. Although the sample is small, there appears to be a correlation between genital tract colonization of GBS in mothers and colonization in their affected newborns.


Adult respiratory distress syndrome (ARDS), sepsis, and extracorporeal membrane oxygenation (ECMO). This report presents data obtained in the care of 830 patients requiring assisted ventilation. When these patients were divided into groups by the severity of their respiratory failure as defined by the duration of ventilatory assistance (greater than 48 hours, less than 48 hours) and level of positive end expiratory pressure (PEEP) required (greater than 5 cm HoH, less than 5 cm HoH), it was found that evidence of concurrent bacterial infection was present in the majority of patients with severe respiratory failure. This finding could not be explained by infection acquired after the onset of respiratory failure. In addition, this analysis demonstrated the important association of active pulmonary infection with the occurrence of barotrauma in these patients. Case analysis of patients subjected to extracorporeal membrane oxygenation has led to the suggestion that underlying sepsis in patients failing to respond to conventional ventilatory assistance similarly limits the usefulness of membrane oxygenator support.


Modifications in the treatment of the multiple injury patient. When burns complicate the management of an otherwise severely injured patient, modifications in treatment must be undertaken for both problems. The unique situation of this type of patient often requires the use of multiple therapeutic interventions which may differ from the usual. The use of controlled hypotension and aggressive surgical burn management was successful in allowing the postponement of vascular surgery in a young male patient with a traumatic aortic disruption and extensive burns resulting from a 60-foot fall and severe thermal burns.


Infections with two dengue viruses in Greece in the 20th century. Did dengue hemorrhagic fever occur in the 1928 epidemic? From contemporary clinical accounts we hypothesized that the 1928 dengue epidemic in Greece may have been an earlier occurrence of dengue hemorrhagic fiver/dengue shock syndrome (DHF/DSS). To study the possibility that two different dengue viruses may have been involved, serums from 62 Athenians alive during the epidemic were examined for dengue antibodies; 73 per cent showed evidence of prior dengue infection. Monotypic neutralizing antibodies were found to two different dengue viruses, types 1 and 2. A large proportion of the sampled population had evidence of two or more past dengue infections. Since there is no evidence that dengue viruses have been transmitted in Greece since 1928, during the epidemic a very large number of persons immune to one dengue type must have acquired infections with a secon type. The virological criteria for secondary infection DHF/DSS are thus satisfied. Although DHF/DSS is currently restricted to Asia and the Pacific, the Greek oubreak suggests a biological potential for fatal consequences of dengue infections in Caucasians, particularly the elderly.


In vitro system that synthesizes circular viral DNA of bacteriophage phi X 174. An extract prepared from Escherichia coli cells infected with phi chi 174 bacteriophage was capable of incorporating dTTP into phage-specific DNAs in vitro. The synthesized DNAs were associated with proteins and sedimented with S values of 20, 50, and 90 in a sucrose gradient sedimentation. DNA isolated from 20S material was open circular replicative form (RF), DNA in 50S material was replicative-form DNA with an extended single-stranded viral DNA that ranged up to one genome in length, and DNA in 90S material consisted of circular and linear single-stranded viral DNA of full genome length and single-stranded viral DNA shorter than full genome length. Pulse and pulse-chase experiments indicated that 90S material derived from 50S material.


Transduction of bacteriophage Mu by bacteriophage T1. Phage T1 transduces phage Mu PFU from Mu-lysogenic donor cells to sensitive recipient cells. The efficiency of transduction depends on the chromosomal location of the Mu prophage. T1, therefore, appears to package different regions of the bacterial chromosome with different efficiencies. Although T1 transduces bacterial markers with different efficiencies, there is no direct correlation between the efficiency of transduction of a bacterial marker and the efficiency of transduction of Mu PFU from donor cells with the Mu prophage located in that marker.


Isolation of specialized lambda transducing bacteriophages for flagellar genes (fla) of Escherichia coli K-12. Specialized transducing lambda phages carrying the region III flagellar genes (fla) of Escherichia coli K-12 were isolated by a new method. A strain carrying both a cryptic lambda prophage near the his genes and a deletion of the attlambda gene was used as a starting strain. The lysogen of lambdacI857pga18-bio69 was isolated in which the prophage was integrated within the lambda cryptic genes by means of recombination with the residual lambda DNA. The strains with deletions starting within the prophage and ending in these fla genes were selected from among the heat-resistant survivors of the lysogen. They were then infected with heat-inducible and lysis-defective lambda phages and, thus, specialized transducing phage lines for hag and fla were obtained. High-frequency transfer lines of rare phages carrying the fla genes were isolated by inducing a strain carrying a heat-inducible lambda prophage near the his genes and selecting by transduction of a fla deletion strain. Preliminary characterization of these transducing phages is also reported.


Location of the ss--mutation of bacteriophage T7 in genes 10, the structural gene for the major capsid protein. T7+ phage are unable to plate on a strain of Shigella sonnei D2 371-48. Spontaneous phage mutants arise (ss--mutants) that are able to plate on this strain of Shigella. We have shown by complementation studies and genetic crosses that the ss--mutation maps in gene 10, the structural gene for the major protein of the capsid. This finding implies that the gene 10 protein may interact with a host protein during phage development and that the abortive infection of T7 observed in S. sonnei D2 371-48 with T7+ phage may be a defect in head morphogenesis. Our studies also reveal that various T7 strains commonly contain deletions in nonessential regions. T7 ss--mutants selected after growth of T7+ on Shigella D2 371-48 often acquire a deletion in the 0.7 gene that is not necessary for the ss--phenotype. Finally, we have found a new nonessential region of the T7 chromosome that is located between 33 and 35.5% of the T7 genome length.


Percutaneous biopsy of renal transplants using ultrasonographic guidance. The transplanted kidney is an ideal organ for study by ultrasonic methods because it is located superficially in the iliac fossa and it is free of superimposed echoes from the intestines. Therefore, it is possible to localize the transplant accurately, measure the distance from the skin to the surface of the kidney and determine with precision the angle of the intended pathway of the needle.


The clinical significance of sulfonamide disk susceptibility testing. A prospective double-blind study in a homogeneous population and according to a uniform protocol documents the value of sulfonamide susceptibility testing, as performed by the disk-agar diffusion method, for the management of chronic urinary tract infections.


Talc granuloma presenting as a testicular mass. A case of a benign talc granuloma in a 3-year-old boy is reported. Surgical wound contamination with various lubricant dusts from surgeons' gloves and resultant granuloma formation is a well known phenomenon. However, there are few reported cases of peritesticular masses owing to talc granulomas.


Ureteropyelostomy and ureteroneocystostomy in renal transplantation: postoperative urological complications. Ureteropyelostomy offers a suitable alternative to ureteroneocystostomy as a primary method of urinary reconstruction after renal transplantation. In a total of 108 such procedures 12 urological complications occurred. Only 1 patient had graft loss directly attributable to these complications.


Commentary on the immunobiology of bladder cancer. This review is an attempt to integrate the conflicting studies that constitute the literature on immunobiology of bladder cancer and to indicate areas that need further investigation. It is clear that immune reactivity directed at antigens expressed on bladder cancer cells and general host immunologic competence decline with progressive tumor growth. This immunodepression correlates with patient prognosis, even within a given stage of disease. The mechanisms underlying this decline in host immune reactivity are poorly understood. A better understanding of these mechanisms may provide fundamental insight into the host-tumor relationship that possibly could be exploited for preventive, diagnostic and therapeutic purposes.


Urinary fistulas after renal transplantation. Urinary fistulas developed in 18 of 200 renal transplants (9 per cent). Twelve fistulas healed after conservative treatment and 6 were repaired by an operation. Our repair technique included constructing a bladder flap for patching, incorporating the fistula into the bladder and suturing the bladder directly to the renal pelvis.


Comparison of intermittent catheterization and supravesical diversion in children with meningomyelocele. A comparison in terms of renal preservation as evidenced by urography is made between children with meningomyelocele on intermittent catheterization and children who had undergone diversion. Followup in the former group is only 2 1/2 years but at this time those patients on catheterization are doing as well and probably better than children who had undergone diversion and the quality of life for them is better.


The role of intermittent catheterization in the management of children with myelomeningocele. Of 114 patients with myelomeningocele 44 (40 per cent) have no significant urologic disability. Intermittent catheterization has been used in 51 children and was helpful in 13 (26 per cent). Cystometrograms and urethral pressure profiles have been useful as predictive parameters. Causes of intermittent catheterization failures are discussed.


Oxybutynin chloride combined with intermittent clean catheterization in the treatment of myelomeningocele patients. Oxybutynin chloride has been with intermittent clean catheterization to achieve urinary continence in 21 of 25 myelomeningocele patients (84 per cent success rate). The regimen, results, problems and implications of this new method of treatment are discussed.


The familial liver donor in renal transplantation. A survey is made of the 20-year experience with more than 300 living donors in renal transplantation at the Peter Bent Brigham Hospital. Details are given pertaining to the first transplantation of the kidney from one living person to another in December 1954. The surgical technique, the complications and the results of renal transplantation from a living donor are summarized. The philosophic and psychologic problems encountered are described. A comparison of results summarized by Murray of living, related donor transplants to cadaveric donor transplants is included. This paper constitutes a salute to the familial living donor.


Ureterosigmoidostomy.

Ureterosigmoidostomy. Some historical aspects of ureterosigmoidostomy are described, and various techniques are discussed and illustrated. Our own satisfaction with the results when the procedure is done through the open sigmoid colon is expressed. Ureterosigmoidostomy, which has in some surgeons' hands fallen into disuse, will continue to be used and probably should be used more than it is at present. When ureterosigmoidostomy is done meticulous care is important in producing a long submucosal tunnel with direct anastomosis of the ureter to the bowel. Preoperative bowel preparation is mandatory. Patients who have undergone ureterosigmoidostomy should remain on a low chloride diet indefinitely with an adequate supplement of sodium potassium citrate to diminish the dangers of electrolyte imbalance and hyperchloremic acidosis. Careful postoperative management and followup care are vital to success.


Hypospadias repair.

Hypospadias repair. When we began our association for the purpose of treating hypospadias in the early 1950s we used a tube graft of full thickness skin in a 1-stage procedure. We have made modifications to improve the appearance of the glans and now use 1 of 4 procedures, dependent on the location of the urethral meatus. The procedures are described and our results are tabulated.


The development of the wire-loop resectoscope and the ensuing controversy concerning transurethral versus suprapubic prostatectomy. The development of instruments and machines to make practical the removal of tissue and the controll of hemorrhage transurethrally by use of high frequency current is traced. The vicissitudes of the early users are reviewed and the feuds between them and those who advocated open prostatectomy are recalled.


An aggressive approach to prostatic cancer. During the last 47 years an aggressive approach to the treatment of prostatic malignancies has been maintained at the Duke University Medical Center. Radical prostatectomy, usually by the perineal route and, more recently, by the retropubic technique in conjunction with pelvic lymphadenectomy, has been vigorously applied in patients with diffuse stages A, B and even C diseases. A 5-year survival rate of 80 per cent has been documented. Hormonal therapy has been equally vigorous, most often embracing orchietomy at the time of diagnosis with immediate initiation of estrogen therapy. Patients so treated exhibited a 5-year survival rate of 49 per cent, far greater than the average survival rates usually quoted. Radiation therapy had been used primarily in conjunction with other treatment measures, and multiple drug chemotherapy is now being used in patients who demonstrate disseminated and progressive disease, refractory to other measures previously instituted. By maintaining flexible combinations of all modalities of therapy our overall 5-year survival rate of 50 per cent has been achieved with all patients exhibiting all stages and classifications of prostatic carcinoma.


Studies of introital colonization in women with recurrent urinary infections. IX. The role of antimicrobial therapy. To determine if antibiotics used in the treatment of urinary infections alter introital gramnegative carriage after termination of therapy we analyzed 254 cultures obtained between episodes of bacteriuria in 14 women with recurrent urinary infections. Cultures obtained within the first 30 days after termination of therapy were compared to all subsequent cultures. Introital carriage in women with recurrent urinary infections was compared to 416 consecutive introital cultures from 31 control women resistant to bacteriuria. In women with recurrent bacteriuria introital colonization patterns were similar in incidence and density during the immediate post-treatment period compared to later cultures. Four volunteer controls received tetracycline for 10 days. There was no difference in introital carriage of enterobacteria before during or after tetracycline therapy. Consecutive cultures also confirmed a higher incidence and greater density of vaginal carriage of enterobacteria in patients when compared to similar cultures from women who never had a urinary infection.


T lymphocyte rosette formation after major burns. The ability of lymphocytes to form rosettes with sheep erythrocytes was studied in the acute burn period to quantitate T lymphocytes. Seventeen adult patients with burns of greater than 15% body surface area were studied serially and compared to normal controls. The absolute number of total rosette-forming cells (RFC) averaged 1,229.1 +/- 92.8/cu mm (SEM) for controls. In patients, a comspicuous decrease in absolute number of total RFC occurred at days 3 to 5 postburn (578.0 +/- 144.2/cu mm) (P less than .001) and during days 6 to 10 (799.8 +/- 106.1/cu mm) (P less than .01). Similarly, the percentage total RFC in patients at days 6 to 10 (48.9% +/- 3.3%) was significantly lower than controls (73.9% +/- 0.7%) (P less than .01). The decrease in total RFC did not correlate with the total lymphocyte count or the cortisol level. These findings suggest that T lymphocytes are decreased or have altered rosette-forming ability in the early postburn period and are in accordance with other reports of impaired cellular immunity following burns.


Respiratory managements in patients with shock. Severe respiratory failure may develop in patients following or during various types of shock. A number of mechanisms for the genesis of this syndrome have been proposed. The exact role of each of these etiologies is unknown, but a combination of these and others may be the actual causative factors. Prevention of respiratory failure consists of prompt resuscitation of shock, and prevention of hypoventilation or atelectasis, embolism and infection. Judicious use of oxygen and electrolyte fluids is also important in prophylactic therapy. At the present the principal feature in treatment consists of IPPV with adequate inspired oxygen concentration. If progressive pulmonary deterioration occurs in patients on IPPV, the CPPV may be indicated. Judicious use of CPPV may result in an apparent improvement of shock lung in some instances.


Determination of the left ventricular volume by scinticamera --basic studies and clinical application--. Scintillation angiocardiography was performed around the point of time when the injected RI mainly passed through the left ventricle. Gating permitted the radiophotography of the end-diastolic and end-systolic image of the left ventricle. Critiques of the techniques gave result that the best available image could be obtained when the cumulative count was 20,000, aperture setting 4/5 and brightness of cathode ray tube 8. Studies with mechanical model showed that the condition of high-flow, combined with the use of relatively small dose of RI resulted in a marked underetimation of the left ventricular image. Clinical studies was performed using SWI-LVEDV plots, giving the plot of stroke index (SWI) against the left ventricular end-diastolic volume (LVEDV). It was thus formed that the patients with enlarged left ventricles responded to the administration of digitalis with shrinkage of the heart, while the patients with relatively small heart responded to it with increase in SWI at the unchanged level of LVEDV. Studies with systolic time intervals particularly PEP/LVET, showed that relatively high values of PEP/LVET applied to those whose left ventricular SWI-LVEDV plots were located low and to the right, while relatively low values of PEP/LVET applied to those patients whose SWI-LVEDV plots were located high and towards the left, indicating relatively good LV performance.


Decreased early diastolic dV/dt in ischemic heart disease observed by ECG-gated radiocardiography. (1) ECG-gated RCG were performed in 127 cases, among them 30 were normal controls, 35 were hypertensives, 16 were ischemic heart diseases, 8 were cardiomyopathies and the rest of 38 cases were of miscellaneous cardiac diseases. (2) ECG-gated RCG were performed consecutively after the preceding RCG. Parameters calculated from RCG and ECG-gated RCG were evaluated together in terms of the sensitivity to discriminate the ischemic heart diseases from the control subjects. Among the parameters evaluated, peak diastolic dV/dt was the most sensitive index to discriminate the ischemic heart group from the control group. PDdV/dt of 20 ml/100 msec/m2 was considered to be a borderline between two groups. (3) Principal component analysis of 12 variables in 127 cases demonstrated that the peak diastolic dV/dt was closely correlated with the cardiac pump function, but was also well related to the ischemic heart factor. (4) ECG-gated RCG following the preceding RCG is a sensitive method to discriminate the ischemic heart disease from the control group.


Occurrence of disseminated intravascular coagulation (DIC) in obstructive jaundice and its relation to biliary tract infection. Coagulation studies were done on 78 consecutive cases of obstructive jaundice with or without biliary tract infection. Among 26 cases with biliary tract infection 20 cases showed no bleeding tendency but remarkable hypercoagulability with decreased fibrinolytic activity. Other six cases developed diffuse bleeding tendency in addition to the signs of hypotension and multiorgan dysfunction such as oliguria, respiratory distress and mental confusion. Most showed marked coagulation defects characterized by thrombocytopenia, decreased fibrinogen, antithrombin III and plasminogen levels and narrowing of maximal amplitude in thrombelastogram as well as the increase of fibrin degradation products and positive soluble fibrin monomer complexes. All except one died and three cases were autopsied. In two cases postmortem examination revealed multiple fibrin thrombi in lungs and other organs. A cause of the development of bleeding tendency in obstructive jaundice presently observed may likely to be due to the occurrence of disseminated intravascular coagulation (DIC), i.e. hypercoagulability caused by the biliary tract infection is responsible.


[Alpha 1 antitrypsin deficiency].

[Alpha 1 antitrypsin deficiency]. Alpha-1-antitrypsin (alpha-1-AT) is a potent protease inhibitor. Its deficiency predisposes to serious diseases such as "neonatal hepatitis" and "obstructive pulmonary emphysema". Due to the existence of multiple codominant alleles at one single locus, there are several genetic variants from alpha-1-AT. In homozygous persons the protease inhibitor type (Pi type) MM is prevailing, in heterozygous persons the Pi types MZ and MS. So far one knows at least 24 different alleles. Their phenotypes differ as well in their electrophoretic position as in the protein concentration of the serum. Pi type MM guarantees a normal concentration of alpha-1-AT in the serum, whereas Pi type ZZ causes of serious alpha-1-AT deficiency which bears a particularly high risk of disease.


[Reye syndrome (author's transl)].

[Reye syndrome (author's transl)]. Reye's syndrome is characterized by severe encephalopathy and fatty infiltration of the liver. Probably this is a polyetiological syndrome. In most cases the disease is preceded by influenza B or varicella infection. As known to date, damage to mitochondria is the essential feature. Therapy is symptomatic. A review is given of the clinical symptoms and of the many unsolved problems of pathogenesis.


[Studies on the question of a possible autoregulation of insulin secretion (author's transl)]. In eight normal weight healthy volunteers it was proved by C-peptide assay whether high concentrations of insulin in serum would suppress the secretion of insulin in an autoregulative manner. Insulin combined with glucose, therefore was given intravenously as a single bolus injection as well as an infusion over a period of two hours. Moreover it was proved whether high levels of insulin would suppress the reactivity of the B-cell to sulphonylurea administration. It was demonstrated that the secretion of the B-cell is regulated only by the concentration of the blood sugar, but not by the level of serum insulin. Likewise the stimulation of the B-cell by sulphonyl-urea administration is not suppressed by high concentrations of insulin.


[Concentration of C-peptide in correlation to kidney function (author's transl)]. Recently, the radioimmunological determination of C-peptide came into interest because of the jugdement of the remaining function of the islet apparatus in insulin-dependent diabetics. As the degradation of C-peptide preferably takes place in the kidney we performed an intravenous glucose load in 32 patients with kidney diseases. The following results were obtained: 1. In patients with a healthy carbohydrate metabolism a clear correlation exists between the concentration of creatinine on the one hand, the creatinine-clearance and the fasting C-peptide concentration respectively the measured amount of C-peptide on the other hand. 2. The more advanced the renal insufficiency the better is the correlation between the parameter of the kidney function and the C-peptide concentration. 3. In diabetic patients there shows to be no clear correlation between the C-peptide levels and the kidney function. --In insulin-dependent diabetics the amount of C-peptide is only of diagnostic use if the renal function is well known.


Investigation of the histocompatibility of the NYA:NYLAR mouse colony by skin grafting. The histocompatibility status of the Nya:NYLAR mouse colony was studied by exchange of skin grafts between female mice. The colony had been divided into two portions since 1962, a larger, outbred stock (Nya:NYLAR), and a smaller, inbred strain (NYLR/Nya). The results of skin graft exchanges between mice of the inbred strain indicated that they were skin-compatible. There was weak skin-incompatibility within the outbred stock and between this stock and the inbred strain, and strong skin-incompatibility between the outbred stock and outbred Webster Swiss mice.


Current status of the therapy of advanced renal carcinoma. A preclinical murine renal model system is available for evaluation of the effectiveness of therapeutic agents. Clinical review reveals that objective response rates to hormonal agents reported since 1971 have been substantially lower than those reported prior to 1971. Many chemotherapeutic agents have not been adequately evaluated clinically or with the experimental model. Vinblastine to date may be the most active single agent with a 25% objective response rate. At the present time, it is uncertain whether combination chemotherapy or hormonal chemotherapy add to response rates obtained with single agents. There is some evidence that immunotherapy may be of some adjuvant benefit and, thus may provide an additional avenue of investigation.


Carcinogenic and possible mutagenic effects of stilboestrol in offspring exposed in utero. The relevant literature has been reviewed and it appears that there is no doubt that in utero exposure to stilboestrol is related to the eventual development of vaginal adenocarcinoma in young females. Although cancer of other types are also prevalent, the incidence is not great and may be related to other types of drugs taken during pregnancy. The review attempts to correlate the genetic effects with the abnormal development, but on account of the scarcity of data such correlation is not possible. Further work on the genetic effects and the development of the lower genital tract in the female needs to be carried out.


Pathogenic factors involved in renovascular hypertension. State of the art. The complex hormonal action of angiotensin II in the long-term control of blood pressure or sodium metabolism, or in renal hypertension, is not completely understood. Structure-activity relations with analogues of angiotensin II gave information about the functions responsible for pressor and myotropic response in the molecule that led to the synthesis of competitive antagonists of this hormone. These antagonists, however, show variable agonist/antagonist ratios in different species or different tissues of the same species. This fact necessitates further work to induce tissue specificity. Although des-Asp1-angiotensin II ("angiotensin III") has been recognized as a hormone, its exact role in the biosynthesis of aldosterone is yet to be discovered. The antagonists such as des-Asp1-[Ile8]-angiotensin II or des-Asp1-[Thr8]-angiotensin II have provided important leads in this direction. Many of the biologic effects of angiotensin I have been attributed to its conversion to angiotensin II by the converting enzyme. Recent investigations indicate that angiotensin I itself may play a direct role; however, most of these studies were carried out by inhibiting the converting enzyme activity with peptides obtained from the venom of Bothrops jararaca. Since these peptides also potentiate bradykinin action, the observed biologic activities could be caused by either angiotensin I or bradykinin. Bsides, converting enzyme is no longer thought to be a single enzyme and its nature varies from species to species and from tissue to tissue in the same species. Renin inhibitors related to renin substrate or pepstatin are not freely soluble in plasma and are not effective under physiologic conditions. This points to the importance of renin inhibitors isolated from kidney or other natural sources. Thus, although the renin-angiotensin system appears to be an integral part of the problem of hypertension, characterization of various converting enzymes, roles of extrarenal renin, isorenin, tonin, and brain-renin, and the involvement of other humoral, neurogenic, and immunogenic factors should be pieced together to get a clear picture of the hypertension problem.


Role of naturally occurring vasoactive principles in hypertension. State of the art. Hypertension may result from excessive activity of one or more components of the blood pressure-elevating system. These include the adrenergic nervous system, the renin-angiotensin axis, and mineralocorticoids (aldosterone), which potentiate each other, reinforcing their effects on renal hemodynamics and electrolyte transport and thereby affecting extracellular fluid volume, vascular tone, and reactivity. We consider of no less importance in the genesis of hypertension the failure of one or more components of the blood pressure-lowering system: the kallikrein-kinin system, prostaglandin, or one or more lipids associated with the renomedullary interstitial cells. As a corollary of this hypothesis, if one assumes tonic activity of these opposing blood pressure-regulating systems, in the case of a deficiency of the vasodepressor system, hypertension should result. That is, unopposed activity of the pressor system should be sufficient to increase blood pressure without an increase in the "basal level" of its activity. Hypertension, then, may be considered to result from either uncompensated deficiencies or excesses, which may be relative or absolute, of one or more components of the vasodepressor and vasopressor systems.


The end-stage renal disease networks: an attempt through Federal regulation to regionalize health care delivery. Recognizing the needs of patients with end-stage renal disease (ESRD), the federal government passed legislation in 1972, establishing Medicare coverage for 92 per cent of the diseased victims regardless of age. However, federal health planners did not in many areas, take into account what was already being accomplished on the state and local level. As a result, the implementation of the provision has created complications of its own. This case study of New Jersey illustrates that until existent informal health delivery networks are incorporated into federal health planning, the taxpayer and health care consumer will not get the most mileage for his or her dollar.


Recent developments in the age-related alteration of enzymes: a review. Recent developments dealing with the age-related alteration of enzymes have been examined. Since the last review of this field, three enzymes have been purified to homogeneity from young and old nematodes (enolase, phosphoglycerate kinase, triosephosphate isomerase) and one from the liver of young and old rats (superoxide dismutase). In all cases except for triosephosphate isomerase, the enzymes from old animals show a reduced catalytic ability compared to those from young animals. In addition, new reports of increases in the amount of altered enzymes in late-passage cells in tissue culture have appeared, though contrary evidence has also been published. The data from these and other papers are compared and discussed. Possible explanations for the alteration of enzymes include error theory, substitution of individual amino acids and conformational change without sequence change. Though final conclusions cannot be made, the evidence favors the latter idea to explain the presence of altered enzymes in old animals.


Experimental antitumor agents from plants, 1974-76. The published literature in the areas of antitumor agents and cocarcinogens, for the years 1974-6 is reviewed. Some of the chemical classes of compounds discussed include tthe sesquiterpene lactones, diterpenes of the phorbol and simaroubolide types, triterpenes of the withanolide and cucurbitacin types, lignans, monomeric and dimeric indole alkaloids, isoquinoline alkaloids, Cephalotaxus alkaloids, camptothecin and the maytansinoids. Emphasis is placed on the isolation, synthesis, biosynthesis and biological activities of the most promising entities.


Glucose kinetics in dogs following a lethal dose of endotoxin. The effects of a lethal dose of Escherichia coli endotoxin on the glucose kinetics and cardiovascular responses of conscious dogs were studied. The plasma glucose level fell steadily after endotoxin administration to severely hypoglycemic levels preterminally. The fall in plasma glucose appeared to be due primarily to an increased tissue uptake in the early phase, whereas decreased hepatic glucose output also contributed in the later, preterminal phases.


Mechanisms of the adjuvant effect of nystatin on in vitro antibody response of mouse spleen cells: indication of nystatin as a B-cell mitogen and as a stimulant for polyclonal antibody synthesis in B cells. Adjuvanticity of nystatin, one of the polyenic antifungal antibiotics having as its primary target the membrane sterol of eukaryotic cells, was investigated by examining its effect on several functions of mouse spleen cells relevant to immunological phenomena in vitro. Nystatin was found to stimulate significantly DNA synthesis in thymus-independent (B) cells but not in thymus-dependent (T) cells. Like the other B-cell mitogens such as bacterial lipopolysaccharide (LPS), nystatin elicited nonspecifically polyclonal antibody synthesis in mouse spleen cell cultures, and also restored antibody response of T cell-deficient spleen cells of congenitally athymic nude mice to heterologous erythrocytes (RBC; thymus-dependent antigen). Thus, nystatin and LPS appeared to cause similar changes in the functions of spleen cells relevant to immunological events. However, antagonism but no additive effect in the adjuvanticity was revealed between the two adjuvants. As an interesting finding, the polyclonal generation of anti-RBC antibody-forming cells (AFC) in the spleen cell cultures by stimulation with B-cell mitogen, i.e., either nystatin or LPS, was not inhibited at all by inclusion of any anti-RBC antiserum, whereas, as is well known, the generation of AFC by stimulation with the antigen was specifically suppressed by the corresponding antiserum, indicating a difference in the genesis between the mitogen-induced AFC and the antigen-induced AFC.


The medical complications of narcotic addiction. I. The major medical complications of needle addiction are reviewed. The problem of overdose, and diseases of the skin, lung and kidney are discussed in detail. The importance and prevalence of infections are emphasized, with special reference to infective endocarditis and viral hepatitis. The work is introduced with a historical review.


Recent developments in the measurement of cardiac output and regional blood flow using indocyanine green and microspheres. Densitometric studies show that the large flow measurement errors and inability to obtain reproducible densitometer calibrations reported with indocyanine green (ICG) in nearly isotonic saline may have arisen from two sources: (a) slowed optical stabilization, and (b) sedimentation of dye aggregates formed in the salt "solutions" of ICG, both of which are avoidable by preparing the dye in water. The assumptions of the widely used regional blood flow measurement technique using radionuclide-labeled microspheres are described. Simultaneous injection of 8mu and 15mu microspheres in turkeys and in dogs demonstrated the existence of at least 8 mu arteriovenous communications (AVCs) in the stomach and intestine, not previously described by this technique, which, in addition to their physiologic functions, may play a role in production of acute gastric mucosal ischemia and erosions. Similar AVCs, producing a lesser degree of "shunting," were also found in the heart. Loss of 8 mu relative to 15 mu microspheres continued with time in the gastrointestinal circulation.


Serum reactions. An analysis of commercial antivenoms and the possible role of anticomplementary activity in de-novo reactions to antivenoms and antitoxins. Infusion of concentrated heterologous serum proteins may precipitate severe reactions in humans. Some of these reactions are not due to a preexisting hypersensitivity to the foreign protein, but rather to the strongly anticomplementary activity (ACA) of the infused protein. The latter mechanism could account for the occasional anaphylactoid reaction seen in a patient who has had no known previous exposure to the foreign protein. Similarly, it could account for the patient who fails to react to a preliminary subcutaneous trial dose, but then collapses when he is infused later with the same preparation. Nine commercial antivenoms from nine countries were examined and most were found to have high levels of anticomplementary activity. Considerable variation was found in both the immunochemical properties and the anticomplementary activity of these antivenoms. Antitoxins of equine origin to the toxins of diphtheria, tetanus and gas gangrene were also found to have high anticomplementary activity. Because of this activity these preparations should always be diluted and infused slowly. Lack of reaction to a test dose will never guarantee an uneventful infusion, and thus all infusions of heterologous immunoglobulin should be undertaken with the greatest caution and vigilance.


Tissue diagnosis in lymphomas.

Tissue diagnosis in lymphomas. An accurate histological diagnosis is sometimes difficult to achieve in the lymphoma patient. This problem is accentuated by incorrect choice of noded for biopsy or mishandling of tissues, either at surgery or during the fixative process. Practical guidelines are suggested to help minimize these problems.


Faculty of Anaesthetists, Royal Australasian College of Surgeons. Foundation and progress. The Faculty of Anaesthetists, Royal Australasian College of Surgeons, was inaugurated in 1952. Events leading to its foundation are described, together with the steps of evolution. An evaluation of its present-day relations with the Australian Society of Anaesthetists and the Royal Australasian College of Surgeons is made, and the extension of its activities to South-East Asia and the community is indicated.


[Bone marrow biopsy investigation: an important prognostic factor in aplastic anaemia (author's transl)]. In thirty-five patients with aplastic anaemia initial bone marrow biopsy investigations were performed. 25 patients showed partial bone marrow aplasia whereas in ten patients complete myeloid aplasia could be demonstrated. Probability of survival was approximately ten times higher in patients with partial than in patients with complete bone marrow aplasia. The results of bone marrow biopsy investigations had a greater predictive value than the degree of diminution of single cellular blood components. Initial bone marrow biopsy investigation facilitates the selection of poor risk patients who should be treated by bone marrow transplantation rather than by conventional therapy.


Synthetic disaccharide-protein antigens for production of specific 04 and 09 antisera for immunofluorescence diagnosis of salmonella. The synthetic disaccharides abequose 1 leads to a 3 mannose and tyvelose 1 leads to a 3 mannose, representative of Salmonella O-antigen 4 and 9 respectively, were covalently linked to bovine serum albumin (BSA) . Antisera from rabbits immunized with these immunogens were used in indirect immunofluorescence assay for the identification of group B (O-antigen 4) and D (O-antigen 9) Salmonella. A total of 1030 enteric bacterial strains were tested, including 207 group B and 55 group D Salmonella. The anti-abequose-mannose-BSA serum correctly identified all Salmonella group B strains tested. The anti-tyvelose-mannose-BSA serum correctly indentified all Salmonella group D bacteria examined with the exception of 11 of 18 Vi-positive S. typhi strains which did not not stain until the Vi-antigen was removed by boiling. Among the 768 strains representing Salmonella other than groups B and D, E. coli, Shigella, Citrobacter, Klebsiella, Enterobacter, Serratia, Proteus, Vibrio, Pseudomonas, Aeromonas, Yersinia, Bacteroides and Fusobacterium only 5 positive reactions were found. These were observed with Y. pseudotuberculosis strains which have the same disaccharide antigenic determinants as Salmonella O-antigen 4 and 9 respectively. The high specificity of the antisera elicited by the synthetic disaccharide-BSA immunogens make them suitable for a specific and rapid identification of Salmonella bacteria belonging to serogroups B and D.


The action of antitumor agents: a double-edged sword? Although cytotoxic immunosuppressive agents play an unquestionably useful role in treating many neoplastic and non-neoplastic disorders, there is accumulating evidence that the toxicity associated with their use is considerable. The therapeutic successes obtained with antitumor agents have led to increases in the life span of cancer patients, but have also provided the opportunity for this toxicity to become manifest. A search of the available literature was carried out, with emphasis on cases in which a malignancy developed in patients following chemotherapy for either neoplastic or non-neoplastic (e.g., renal transplantation, psoriasis) conditions; particular focus was given to the incidence of acute myelogenous leukemia in various groups of Hodgkin's disease and multiple myeloma patients. That patients with nonmalignant conditions treated with cytotoxic immunosuppressive agents are also at increased risk of developing a malignancy raises the possibility that these agents may have oncogenic potential. Therefore, one may be faced with the paradox that the patients benefiting most from chemotherapy may be at highest risk of suffering its consequences.


Moral dilemmas in clinical cancer experimentation. An examination of certain moral tensions that have developed in clinical cancer experimentation is offered. It is argued that the individual partner must be addressed in his singular unique position, rather than as a potential for the future good.


Detection of messenger RNA from the isoleucine--valine operons of Salmonella typhimurium by heterologous DNA-RNA hybridization: involvement of transfer RNA in transcriptional repression. A hybridization assay using Escherichia coli K-12 DNA isolated from the specialized transducing bacteriophage gammaCI857St68h80 dilv was used to examine the rate of synthesis of the messenger  RNA's (mRNA) derived from the isoleucine-valine (ilv) gene cluster of Salmonella typhimurium. In all cases examined, changes in ilv enzyme levels could be correlated with changes in the rate of synthesis of ilv mRNA. Several well characterized regulatory mutants of S. typhimurium had rates of synthesis of ilv mRNA 3 to 8-fold higher than the repressed wild-type strain. The increased rates of ilv mRNA synthesis found in a hisT strain as well as in isoleucyl-and leucyl-tRNA SYNTHETASE MUTANTS, STRONGLY SUGGESTS A ROLE FOR BRANCHED-CHAIN AMINOACYL-TRNA's in transcriptional control.


The transposon Tn1 as a probe for studying ColE1 structure and function. Insertion of the transposable genetic element Tn1 into different sites of plasmid ColE1 results in a number of mutnat phenotypes. Whereas all plasmid examined were present in normal amount, all showed reduced immunity to killing by colicin E1. Of six insertions isolated after conjugation, five fail to produce colicin, are conjugally proficient (transmissible), and map within a 500 nucleotide region of the genome. The other is conjugally deficient, produces colicin normally and maps close to two others with a similar phenotype isolated after transformation. Of four others isolated after transformation, two have similar properties to the original five transmissible plasmids. The other two are nontransmissible and produce colicin. Non-transmissibility is correlated with reduced relaxation complex. Patterns of protein synthesis in minicells by ColE1 and ColE1 :: Tn1 plasmids have been examined: all ColE1 plasmids containing Tn1 show an altered pattern of ColE1 protein synthesis in addition to three presumptive Tn1-specified proteins, one of which is shown to be beta-lactamase. ColE1 :: Tn1 plasmids can be inserted into the conjugative plasmid R64drd11 to form a cointegrate in which ColE1 and Tn1 function can be expressed.


Studies on the bipolar argECBH operon of E. coli: characterization of restriction endonuclease fragments obtained from gammadargECBH transducing phages and a ColE1 argECBH plasmid. The isolation of a new type of gamma transducing phage carrying the bipolar argECBH operon of E. coli K12 is described. The argECBH segment is inserted in the phage in a direction which is opposite from that of previously isolated argECBH-carrying phages. A colE1 argECBH plasmid has been constructed. DNA fragments resulting from digestion of these genetic elements with Eco RI and Hind III restriction enzymes have been characterized by agarose gel electrophoresis and electron microscopy, including hetero-duplex analysis. Two fragments are of special significance for studies on the control of arginine synthesis, one of length 9.8 kilobases carrying the whole argECBH region, the other of length 2 kilobases carrying most or all of the control region between argE and argC.


Analysis of the phase variation in lambda reduced immunity lysogens. Two distinct phases characterized by different levels of immunity that appear in some E. coli strains lysogenic for reduced immunity mutants of bacteriophage lambda are identified as single and tandem double lysogens respectively on the basis of DNA-DNA hybridization experiments and the requirement of the phage xis function for the transition from a single to a double, and of the host recA function for the transition from a double to a single lysogen (in a xis- condition). Rim lysogens with a further increase in immunity, containing some 5 copies of the lambda genome per host genome, have also been observed. It is argued that the different levels of immunity are a direct reflection of the CI gene dosage effect. An unexplained finding is that rim single lysogens yield double lysogens with a frequency of near 1% per generation, whereas cured cells fail to appear even at a frequency 100 times lower.


Complementation of transfer deficient ColE1 mutants. The transfer defect of some ColE1 mutants is complemented by ColE1 or ColK, but not by ColE2. This implies that at least one ColE1-specified protein or RNA is normally needed for ColE1 conjugal transfer. The gene(s) postulated for this function lies within a region whose length is at most 50% of the genome.


Altered dihydrofolate reductase in fol regulatroy mutants of Escherichia coli K12. Three spontaneous fol regulatory mutants contain dihydrofolate reductase molecules which differ in physical properties from enzymes of their parent strains. The enzymes were purified over 100-fold by affinity chromatography and were shown to differ in vitro in thermolability and in affinity for trimethoprim, a competitive inhibitor of the enzyme. These results indicate that some fol regulatroy mutations occur in the structural gene for dihydrofolate reductase.


Defective thymine dimer excision in radiation-sensitive mutants rad10 and rad16 of Saccharomyces cerevisiae. Two rad mutants of yeast, rad10 and rad16, are shown to be defective in the removal of UV-induced pyrimidine dimers since DNAs obtained from irradiated cells following a post-irradiation incubation in the dark still retain UV-endonuclease-sensitive sites. Both rad10 and rad16 mutants are in the same pathway of excision-repair as the rad1, rad2, rad3 and rad4 mutants.


Physical and genetic studies with restriction endonucleases on the broad host-range plasmid RK2. The cleavage map of the plasmid RK2 was determined for the five restriction endonucleases EcoRI, HindIII, BamH-I, SalI and HpaI. DNA has been inserted into several of these sites and cloned in Escherichia coli. Efforts to obtain derivatives of RK2 reduced in size by restriction endonuclease digestion of the plasmid were not successful and indicated that genes required for the maintenance of this plasmid in E. coli are not tightly clustered. An RK2 derivative possessing an internal molecular rearrangement was obtained by transformation with restriction endonuclease digests of the plasmid.


Interaction of super-repressible and dominant constitutive mutations for the synthesis of galactose pathway enzymes in Saccharomyces cerevisiae. Two dominant uninducible mutant alleles in the gal80 locus were identified. The GAL80s-1 and GAL80s-2 mutants showed novel phenotypes in response to the newly isolated GAL81-1 mutant allele, a dominant constitutive mutation linked to the gal4 locus; the GAL80s-1 GAL81-1 strain was inducible and the GAL80s-2 GAL81-1 strain was uninducible. Many galactose positive revertants from the GAL80s-2 GAL81-1 strain were isolated. It was proved that each revertant was due to a secondary mutation either in the gal80 or GAL81 locus, whereas revertants due to mutation at the supposed controlling site for the structural gene cluster of the galactose-pathway enzymes have not been isolated.


Replication of the mini-R1 plasmid Rsc11 and Rsc11 hybrid plasmids. Replication of the multicopy mini-R1 plasmid, Rsc11, is dependent on host replication functions dna A, B, C, E and G but independent of polA1. Chloramphenicol immediately stops its replication. A stable relaxation complex is not formed. Composite plasmids were constructed with Rsc11 and other small replicons like pSC101, ColE1 and mini-ColE1. In all combinations the amount of hybrid plasmid DNA in the cell never exceeds the amount of Rsc11 DNA itself. This leads to varying copy numbers of the hybrid plasmids depending on the size of the second plasmid. Replication of the composite plasmids proceeds probably always under the control of the Rsc11 part although the second replicon is still functional. The composite plasmids are incompatible with both the parent replicons.


Control of ribosomal RNA synthesis in Escherichia coli. II. Ribosomal RNA synthesis in isolated nucleoids. The effect of amino acid-starvation on the transcription in vitro of overall RNA and ribosomal RNA was investigated using nucleoids prepared from the exponentially growing and the amino acid-starved cells of rel+ and rel- strains of Escherichia coli. In this system, the synthesis of RNA is exclusively due to elongation of the chains which have been initiated in vivo. The amounts of overall and ribosomal RNA synthesized per unit of DNA in the nucleoids were analyzed for each preparation. The following observations have been made. (1) The total RNA synthesis per unit of DNA in the nucleoids from the amino acid-starved rel+ and rel- cells was not significantly different from each other. (2) The preferential ribosomal RNA synthesis occurred in the nucleoids from the growing cells; the ribosomal RNA synthesis was restricted in the nucleoids from the starved rel+ cells, while no restriction was observed in the nucleoids from the starved rel- cells. The results suggest that the ribosomal RNA synthesis is regulated at the initiation or less likely elongation level of the transcription. (3) A ribosomal RNA of a discrete size of about 30S was synthesized in the nucleoids. No mature ribosomal RNA species was produced in this system. The 30S RNA is probably a primary transcript of ribosomal RNA genes containing 23S, 16S and 5S mature ribosomal RNA sequences.


Control of ribosomal RNA synthesis in Escherichia coli. III. Cytoplasmic factors for ribosomal RNA synthesis. The ribosomal RNA synthesis in a cell-free system containing the nucleoids and the cytoplasmic fraction prepared from Escherichia coli cells has been investigated. The addition of the "4S" fraction from the cytoplasm to the isolated nucleoids induces RNA synthesis by a new chain initiation. In this system a preferential initiation or rRNA chains occurs. The experimental results suggest that the 4S fraction contains at least two activities, one for releasing RNA-polymerases from the nucleoids, and another for the frequent initiation of rRNA chains. No restriction of the rRNA synthesis has been observed in the nucleoids and the 4S fraction from the amino acid-starved rel+ cells. The rRNA synthesized in the above system is detected at about 23S and 16S rRNA regions.


Restriction endonuclease mapping and mutagenesis of the F sex factor replication region. The plasmids pSC138 and pML31 each contain the EcoRI-generated f5 replicator fragment of the conjugative plasmid F in addition to an EcoRI fragment encoding antibiotic resistance: ampicillin resistance derived from Staphylococcus aureus in pSC138 and kanamycin resistance from Escherichia coli in pML31. We have mapped one HindIII and two BamHI restriction sites in the f5 region of these plasmids and one HindIII site in the antibiotic resistance region of each plasmid. The HindIII site in the Km region of pML31 occurs in the kan gene whereas the HindIII site in the Ap region of pSC138 appears to occur in an area important for the regulation of beta-lactamase production. By means of in vitro recombinant DNA manipulation of plasmids pML31 and pSC138, we have shown that approximately 1.9 X 10(6) daltons of the 6.0 X 10(6) dalton f5 fragment can be deleted without disrupting plasmid stability. In addition, we have used these same techniques to isolate a novel F-controlled Ap plasmid cloning vehicle which contains a single restriction site for each of the enzymes EcoRI, HindIII, and BamHI. This cloning vehicle has been linked via either its EcoRI or HindIII site to a ColE1 plasmid replicon to yield stable recombinants.


Pyruvate kinase mutants of Saccharomyces cerevisiae: biochemical and genetic characterisation. Mutants of Saccharomyces cerevisiae lacking pyruvate kinase (EC 2.7.1.40) are described. These have less than 0.5% of the pyruvate kinase activity of the wild type. All the other glycolytic enzymes are present in normal amounts in these mutants. The mutation is recessive and segregates in diploids as a single gene. Five alleles examined fail to complement one another. Tetrad analysis and mitotic recombination data place the mutation on the left arm of chromosome I distal to cys 1. The majority of single-step spontaneous revertants on glucose regain the enzyme activity fully and this activity appears, by a number of criteria, to be due to the same enzyme present in the wild type. Some of these revertants become nuclear petites. The mutants do neither grow on nor ferment sugars but do grow on ethyl alcohol or pyruvate. Glucose addition to cultures growing on alcohol arrests growth until glucose is exhausted. The steady state rate of glucose utilization is slower than in the wild type. This is associated with the accumulation of as much as 5 micronmoles P-enolpyruvate per g wet weight of cells and proportional amounts of 2-P-glyceric and 3-P glyceric acids. The mutation is believed to involve some regulatory element in the synthesis of pyruvate kinase.


A specialized transducing lambda phage carrying the Escherichia coli genes for phenylalanyl-tRNA synthetase. A lambda phage has been isolated which specifically transduces the Escherichia coli pheS and pheT genes coding for the alpha and beta subunits of the phenylalanyl-tRNA synthetase (PRS). This phage transduces with high frequency (i) several temperature-sensitive PRS mutants to thermoresistance and (ii) a p-fluorophenylalanine resistant PRS mutant to sensitivity against this amino-acid analog. The in vitro PRS activities of such lysogens suggest that the alpha and beta subunits coded by the transducing phage complement the mutant host PRS-subunits in vivo by means of formation of hybrid enzymes. The transducing lambda phages were also used to infect UV light irradiated cells. The SDS-gel electrophoretic analysis of the proteins synthesized in such cells revealed that the phage codes at least for four different E. coli proteins. Two proteins with molecular weights of 94,000 and 38,000 daltons cross-reacted with an anti PRS serum and were thus identified as the beta and alpha subunits of PRS, respectively. A third protein with a molecular weight of 22,000 daltons is identical with the ribosomal initiation factor IF3 (Springer et al., 1977b). The other protein (Mr 78,000) is still unidentified.


Replication of Fpoh+ plasmid in mafA mutants of Escherichia coli defective in plasmid maintenance. A class of F' plasmids, designated Fpoh+, was previously shown to be able to replicate extra-chromosomally on Hfr strains by virtue of carrying the specific site or region poh+ (permissive on Hfr) of the E. coli chromosome (Hiraga, 1975, 1976a). These plasmids were now found to replicate on E. coli mafA mutants (mafA1 and mafA23) that cannot support vegetative replication of F and some other F-like plasmids. The derivatives of Fpoh+ that have lost the poh+ site, on the other hand, failed to replicate on mafA mutants. These mutants harboring Fpoh+ (but not Poh- derivatives thereof) exhibit abnormal cell division and form elongated cells, presumably due to competition between Fpoh+ and the host chromosome for some factor(s) essential for the initiation of DNA replication of the both replicons. It is tentatively concluded that the poh+ site is required for F' plasmids to replicate on mafA mutants as well as on Hfr strains. In view of the fact that the mechanism of inhibition of autonomous F DNA replication in mafA mutants and in Hfr strains are clearly different, the present data seem to provide strong support to the notion that the poh+ region contains the replication origin of the E. coli chromosome.


The use of a novel plate assay in a search for yeast mutants defective in deoxyribonucleases. Methods by which the intracellular enzymes deoxyribonuclease, ribonuclease and protease can be assayed in whole colonies of Saccharomyces cerevisiae on agar plates are described. A search for mutants deficient in deoxyribonuclease has been carried out. Two types of mutant are described. One apparently fails to produce deoxyribonuclease, ribonuclease or protease on agar plates and the other apparently fails to produce deoxyribonuclease and ribonuclease.


Properties of hybrid plasmids, consisting of parts of the mini-R1 factor Rsc11 and ColE1. In vitro joining of the two small multicopy plasmids Rsc11 and ColE1 by a poly dAdT linker resulted in hybrid plasmids, which determine resistance to ampicillin and immunity to colicin E1. Isolation of the plasmid DNA from single colonies revealed that a variety of hybrid plasmids was formed. Cleavage of these plasmids with restriction endonucleases HinII, HindIII, EcoRI, SmaI and BamI and hybridization with ColE1 demonstrated that they contain different parts of the parent plasmids, Rsc11 and ColE1. Their copy number in the cell is between 6 and 15 per chromosome depending on the plasmid. None of these plasmids can replicate in polA mutants. Replication continues in the presence of chloramphenicol. This suggests that replication can only occur from the ColE1 origin and that the replication function of the Rsc11 part is lost. The hybrid plasmids are compatible with Rsc11 but not with ColE1. The comparison of the physical maps of these Rsc11--ColE1 hybrids with their functions allows a partial determination of the location of ampicillin resistance, replication and incompatibility on the Rsc11 genome.


Further temperature-sensitive mutants of Escherichia coli with altered ribosomal proteins. Various alterations in ribosomal proteins were detected in forty-one mutants of E. coli isolated as temperature-sensitive mutants. Out of these, six are new classes of mutants harboring mutations in proteins S3, L5, L7 (L12), L29, L30 and L33. One of them apparently lacks protein L7 of the large subunit. These mutants together with those reported previously (Isono et al., 1976) total one hundred and one ribosomal mutants in thirty different proteins.


Kinetics of ribosome synthesis during a nutritional shift-up in Escherischia coli K-12. The rates of total protein synthesis, polyribosome formation and 70S ribosome accumulation were measured following a nutritional shift-up of Escherichia coli K-12. Changes in ribosome content and distribution during the shift-up were measured by examining the total cellular content of free and polysome-associated ribosomes using a sensitive double isotope labeling method. The kinetics of ribosomal subunit formation and the biosynthesis of subunit protein and RNA species were also defined. The results indicated that a pre-shift population of ribosomal subunits was utilized for the immediate post shift increase in both total and ribosomal-specific protein synthesis. An assembly time for new subunits of about 3 min was observed. The formation of certain ribosomal proteins during the shift suggested that new subunit assembly was limited by the rate of synthesis of particular ribosomal proteins during this growth transition.


Mitochondrial DNA replication in petite mutants of yeast: resistance to inhibition by ethidium bromide, berenil and euflavine. Mitochondrial DNA (mtDNA) replication in petite mutants of Saccharomyces cerevisiae is generally less sensitive to inhibition by ethidium bromide than in grande (respiratory competent) cells. In every petite that we have examined, which retain a range of different grande mtDNA sequences, this general phenomenon has been demonstrated by measurements of the loss of mtDNA from cultures grown in the presence of the drug. The resistance is also demonstrable by direct analysis of drug inhibition of mtDNA replication in isolated mitochondria. Furthermore, the resistance to ethidium bromide is accompanied, in every case tested, by cross-resistance to berenil and euflavine, although variations in the levels of resistance are observed. In one petite the level of in vivo resistance to the three drugs was very similar (4-fold over the grande parent) whilst another petite was mildly resistant to ethidium bromide and berenil (each 1.6-fold over the parent) and strongly resistant (nearly 8-fold) to inhibition of mtDNA replication by euflavine. The level of resistance to ethidium bromide in several other petite clones tested was found to vary markedly. Using genetic techniques it is possible to identify those petites which display an enhanced resistance to ethidium bromide inhibition of mtDNA replication. It is considered that the general resistance of petites arises because a product of mitochondrial protein synthesis is normally involved in facilitating the inhibitory action of these drugs on mtDNA synthesis in grande cells. The various levels of resistance in petites may be modulated by the particular mtDNA sequences retained in each petite.


Temperature-sensitive respiratory-deficient mitochondrial mutations: isolation and genetic mapping. In order to find new genetic loci and functions on the yeast mitochondrial DNA, especially mutations affecting the mitochondrial protein synthesis apparatus, temperature sensitive mutants have been isolated after MnCl2 mutagenesis and mitochondrial and nuclear mutants classified according to their pattern of recombination with three rho- tester strains. Eighteen cold- and heat-sensitive respiratory deficient mitochondrial mutants have been isolated and localized on the mitochondrial genome by deletion mapping using 113 rho- strains. Eight of them appear to represent new loci, among which some are probably mutations of the tRNA and rRNA genes.


Cold-sensitive growth of a mutant of Escherichia coli with an altered ribosomal protein S8: analysis of revertants. 26 cold-resistant revertants of a cold-sensitive Escherichia coli mutant with an altered ribosomal protein S8 were analyzed for their ribosomal protein pattern by two-dimensional polyacrylamide gel electrophoresis. It was found that 16 of them had acquired the apparent wild-type form of protein S8, one exhibits a more strongly altered SC than the original mutant and two revertants regained the wildtype form of S8 and, in addition, possess alterations in protein L30. The ribosomes of the residual revertants showed no detectable difference from those of the parental S8 mutant. The mutation leading to the more strongly altered S8 was genetically not separable from the primary S8 mutation; this indicates that both mutations are very close to each other or at the same site. The structural gene for ribosomal protein L30 was mapped relative to two other ribosomal protein genes (for proteins S5 and S8) by the aid of one of the L30 mutants: The relative order obtained is: aroE....rpmD(L30)....rpsE(S5)....rpsH(S8)....rpsL(S12). The L30 mutation impairs growth and ribosomal assembly at 20 degrees C and is therefore the first example of a mutant with defined 50S alteration that has (partial) cold-sensitive ribosome assembly. A double mutant was constructed which possesses both the S8 and the L30 mutations. It was found that the L30 mutation had a slight antagonistic effect on the growth inhibition caused by the S8 mutation. Thus the L30 mutants might have possibly arisen from the original S8 mutant first as S8/L30 double mutants which was followed by the loss of the original S8 lesion.


Mapping of mutation tsm-8 with respect to transfer RNA genes on the mitochondrial DNA of Saccharomyces cerevisiae. A precise localization of the tsm-8 mutation in relation to the transfer RNA genes has been attempted by rho- deletion analysis. The data show that the tsm-8 mutation is in close proximity to the isoleucyl transfer RNA gene. However, it is not yet possible to decide whether the tsm-8 mutation is within this transfer RNA gene.


[Do modern surgical methods in the treatment of retinal detachment represent progress?]. The results of plombage and encircling procedures in 417 eyes operated on for retinal detachment are compared with the rate of success obtained by lamellar or penetrating scleral resection in the past.


Population and family studies to demonstrate Ir genes: HLA haplotype in atopic allergy. 47 normal healthy controls and 45 atopic individuals together with five families with allergic diathesis and sensitive to mite antigen were typed for 13 HLA antigens. In general population, HLA-A1 and HLA-A8 were found to be higher than in normal controls. But the difference was significant at a level of p 0.05 after correction was made for the number of HLA antigens used. In one family, an HLA-A1 -B8 haplotype seems to be linked to the manifestation of atopic disease. The pattern of association of an HLA-A9 -B7 haplotype with the disease process in another family indicates that multiple genes may be involved in the manifestation of a variety of allergic disorders. Although more than one HLA haplotype of HLA-associated Ir gene for one antigen have been postulated, we still do not have enough evidence to suggest linkage between HLA haplotypes and atopic allergy. Further studies utilizing mixed lymphocyte culture and carefully matched control with a large number of subjects and more families are in progress to study the genetic basis of allergic diseases.


The use of rec-bacteria for testing of carcinogenic substances. A series of rec-Escherichia coli strains were tested for their sensitivity to four known carcinogenic compounds by examination of a zone of inhibited bacterial growth around a central well containing the test chemical. The mutants recA-, recB-, recC-, and recA- recB- recC- were all more sensitive to the mutagens than the parental strain AB1157. The recB- recC- strain was examined with a larger series of compounds and was found to respond to many of the substances in a similar way as the Salmonella typhimurium strains of Ames but with some notable exceptions. Nitrosamines, with rat liver microsomal activation, could be detected at lower levels and a group of aromatic amino compounds failed to react with these rec-E. coli. An unusual feature of these rec-mutants is their sensitivity to mixtures of nitrosamines and 2-acetyl amino-fluorene in the absence of microsomal activation.


Relative efficiencies of a series of square-planar plantinum(II) compounds on Salmonella mutagenesis. Twelve Pt(II) compounds have been tested for mutagenicity on Salmonella typhimurium (strain TA 100). Very high mutagenic activities were found for the cis derivatives. A correlation is suggested between these results and a formerly described model of chemical reactivity towards DNA, according to which cis derivatives from intra-strand chelates with guanine. A smaller activity was found with monodentate complexes with DNA.


Mutagenicity of anti-cancer nitrobenzofuroxans. Anti-leukaemically active benzofuroxans were tested for mutagenicity in Salmonella typhimurium. Mutagenicity was not found to be correlated to the previously established anti-leukaemic activity. One anti-leukaemically inactive compound after exposure to liver microsomal enzymes proved the most mutagenic of the derivatives for TA100, whereas after similar treatment, the mutagenicity of the most potent anti-leukaemic compound was reduced. All twelve derivatives tested were mutagenic in a base-substitution strain which was defective in excision-repair and also carried a plasmid-linked repair deficiency. Mutagenicity of five dervatives was undetectable in strains proficient for one or the other of the above repair pathways. Nine of the benzofuroxans could also be detected as mutagens in the frameshift tester strain TA98.


The mutagenic action of nitroimidazoles. II. Tinidazole, ipronidazole, panidazole and ornidazole. The 5-nitroimidazoles tinidazole (Fasigyn), ipronidazole (Ro-7-1554), panidazole and ornidazole (Tiberal, Ro-7-0207) in concentrations of 0.02--1 mM per liter increased the mutation frequency of Klebsiella pneumoniae. Escherichia coli K12 and Citrobacter freundii to streptomycin resistance, including streptomycin dependence, in Luria and Delbrück's fluctuation test. At low concentration (0.1 mM), the increase of the mutation frequency caused by each compound was nearly the same, i.e. 3--4 times the spontaneous mutation frequency. At higher concentrations, considerable differences between the mutagenic activities of the compounds occurred.


Resistance and mitotic instability to chloroneb and 1,4-oxathiin in Aspergillus nidulans. Mutants resistant to two fungicides, chloroneb (1,4-dichloro-2,5-dimethoxybenzene) and vitavax (2,3-dihydro-5-carboxanilido-6-methyl-1,4-oxathiin) were spontaneously obtained from a strain of Aspergillus nidulans with frequencies of 12.5 and 1.1 respectively, in 10(8) conidia. One chloroneb-resistant mutant (Chl 1) segregated as a single gene and was mapped in linkage group IV. It also caused a partial dependence of the strain on the fungicide and was semi-dominant. The mutant resistant to vitavax (Vit 1) also segregated as a single gene and was dominant. Both fungicides altered the instability of diploid and duplication strains. Chloroneb mainly increased haploidization, and vitavax reduced the mitotic recombination in diploids. Chloroneb increased the instability of duplication strains, and vitavax reduced such instability. The possible mode of action of such fungicides affecting stability is discussed.


Detection of mutagenic activity in particulate air pollutants. Ames's strains of Salmonella typhimurium were used to evaluate the mutagenic activity of airbone particulate materials collected at six different points in the industrial area of Ohmuta and the residential area Fukuoka. Tests were done in presence of rat-liver S-9 fraction isolated from rats that had been treated with Aroclor 1254. When the number of revertant colonies per plate was plotted against the amount of methanol extract of particulate air pollutants, using strain TA98, approximately linear relationships were observed for active samples. Generally, mutagenic activity of the samples increased in proportion to the density of air pollutants. In our system, 38--349 microng of methanol extract, from 0.225--4.51 m3 of air from the factory districts in Ohmuta City gave 100 his+ revertants per plate. On the other hand, 54--2300 microng of air pollutants, from 1.29--14.1 m3 of air from the residential districts in Fukuoka City, gave a comparable activity. Every sample from each area had mutagenic activity. Chemicals in air pollutants were fractionated by alumina column chromatography and identified by gas chromatography and mass spectrometry. More than 28 compounds, including 12 unknown substances were identified as polycyclic hydrocarbons. Twelve of these compounds are already known to be carcinogens and to induce reversions to histidine independence in strain TA98 of Salmonella.


Mutagenicity of polycyclic hydrocarbons. II. Monitoring genetical hazards of chrysene in vitro and vivo. Mutagenicity tests were performed with chrysene in the Salmonella/microsome test, NMRI-mice oocytes, bone-marrow cells and spermatogonia of Chinese hamsters. Only in mice oocytes was a weak but significant increase of structural chromosome aberrations observed. Correlations were found between weak carcinogenic and observed weak mutagenic activities of chrysene in vitro and in vivo.


Marijuana exposure in vivo and human lymphocyte chromosomes. Sequential chromosome examinations of peripheral lymphocte cultures were carried out on 21 adult male volunteers who smoked natural blend marijuana cigarettes containing about 1%, 2%, or no delta9-THC. For a limited number of subjects, blood samples from a single venipuncture were cultured independently in two cytogenetic laboratories, and later the slides were exchaged for re-analysis. There were significant differences between laboratories in the absolute break frequencies recorded. These inter-laboratory differences were demonstrated for both techniques of cell culture and metaphase analysis. Neither laboratory found a statistically significant increase in break frequencies asssociated with marijuana smoking. The present study, therefore, failed to detect a measurable effect of marijuana smoking on chromosomal aberrations in subjects experienced in the use of the drug.


Mutagenic studies with acrylonitrile.

Mutagenic studies with acrylonitrile. The mutagenicity of acrylonitrile (vinyl cyanide, propenenitrile) has been demonstrated in the Ames Salmonella typhimurium/liver microsome assay system. Acrylonitrile, in the presence of a mouse liver homogenate produced mutations in the TA 1535, TA 1538 and TA 1978 strains. Exposure of the bacteria was achieved by spotting the acrylonitrile on a "lawn" of salmonella, by shaking a reaction mixture consisting of bacteria, liver homogenate and acrylonitrile, and by exposing the homogenate and bacteria to an atmosphere containing the acrylonitrile. Mutagenesis by this latter method was observed at exposures as low as 57 ppm, less than three times the TLV of 20 ppm that is designated in the United States.


Mutagenic activity of amino acid pyrolyzates in Salmonella typhimurium TA 98. Pyrolyzates of 25 amino acids and 5 indole derivatives were tested for mutagenicity in the histidine-requiring mutant Salmonella typhimurium TA 98. Significant mutagenic activity was detected with pyrolyzates of most of the amino acids. These pyrolyzates required a liver microsomal fraction, as representative of mammalian metabolism, to be detected as mutagens. Among the pyrolyzates tested, the highest mutagenic activity was observed with that of L-tryptophan. As little as 10 microgram of the pyrolyzate of L-tryptophan had detectable mutagenic activity toward TA 98. The optimal pyrolysis temperatures for the formation of mutagenic products were shown to be 500 degrees C for L-tryptophan and 600 degrees C for the other amino acids. The results from pyrolyses of some indole derivatives suggest that an amino group at the alpha-position to the carboxyl group of L-tryptophan plays an important role in the formation of mutagens.


Mutagenicity tests on anthelmintics: microsomal activation of viprynium embonate to a mutagen. Eight anthelmintic preparations readily available in Australia were tested for mutagenicity in the Salmonella typhimurium test system. A slightly modified version of the procedure recommended by Ames et al. [2] was adopted, in that the test samples were placed in "wells" cut out of the agar of a plate previously seeded with the appropriate tester strain. Addition of a mixture of rat liver microsomal enzymes and appropriate co-factors ("S-9 mix") to one of the two wells on a single plate allowed a possible requirement for metabolic activation to be recognised. Using this procedure, viprynium embonate was found to be non-mutagenic. It was however, activated by the rat liver microsome preparation to a mutagen capable of causing both base-pair substitution (detected with strain TA100) and frameshift (detected with strain TA98) mutations. The other seven compounds tested all gave negative results in this system.


Mutagenicity of nitrovin--a nitrofuran feed additive. Nitrovin, a nitrofuran feed additive, is shown to be directly mutagenic in Salmonella typhimurium TA 98 and TA 100 between 0.1 and 2.5 microgram per plate (0.09--2.3 micrometer). Addition of a rat-liver homogenate reduces the mutation rates. Nitrovin inhibits growth of the same bacteria in suspension cultures at concentrations above 0.09 micrometer.


Relation between chemical constituents of tobacco and mutagenic activity of cigarette smoke condensate. Mutagenic activities of cigarette smoke condensate were assayed in the presence of S-9 Mix using Salmonella typhimurium TA 98. The results were examined in relation to chemical data of tobacco leaves. Among the nitrogenous constituents examined, the contents of total nitrogen and protein nitrogen and the soluble nitrogenous fraction were positively and significantly related to an increase in mutagenic activity of the smoke condensate, whereas nicotine and nitrate were not important in contributing to mutagenic potency of such condensates. The age of tobacco leaves influenced the mutagenic potency of the condensate, which was lowest in leaves from the lower stalk position and increased with ascending leaf position on the stalk. Smoke condensate from tobacco with higher sugar content resulted in lower mutagenic activity. The present results, together with the previous study on the mutagenicity of the amino acid pyrolyzates, suggest that potent mutagens in cigarette smoke condensate are nitrogen-containing compounds, which may be formed from proteins and amino acids during the burning of a cigarette.


Is an early calcium flux necessary to stimulate lymphocytes? Concentrations of concanavalin A or the calcium ionophore A23187 that are optimal for the transformation of pig or mouse lymphocytes do not normally cause a measurable increase in calcium influx compared with unstimulated cells. If the cells are treated with the mitogens in conditions where a measurable increase in calcium influx occurs, no stimulation of the cells can occur while the flux is maintained. If an early influx of extracellular calcium is necessary for stimulation, then a much smaller increase in the total concentration of cellular calcium than reported previously is sufficient to allow the entry of lymphocytes into the cell cycle.


[Psychiatry and mental health in the Institutp de Seguridad Social para los Trabajadores del Estado. Philosophy of its development]. In 1972, prepaid medical care for government employees provided by their social security institute, ISSSTE, was reorganized. A division of planning and technical standards was established, within which a Department of Psychiatry was included. Psychiatric care was restructured at three levels: psychiatric hospital, psychiatric OPD at clinic and hospital level and a pilot program in community psychiatry. A three-year psychiatric residency program was established, in addition to participation in other postgraduate, in-service training and monographic courses. Systematic research was started, as well as a publications program, working relationship with other institutions and societies were enhanced. A descriptive example is Child Psychiatry. Most frequent diagnoses are reviewed, and development of services is followed in relation to pediatric departments.


Reorganizations of health agencies by local government in American urban centers: what do they portend for "public health"? The major health agencies of local urban governments--public health department and public hospital--are often dismissed as "irrelevant" or attacked as "unresponsive". The rush to reform through reorganization hasn't often addressed the underlying problems of an eroded tax base, an indifferent federal policy, and intra-professional disputes. Newer corrective steps are called for if complex demands for public and personal service are to be met.


Licensure of foreign medical graduates: an historical perspective. Today, one out of five practicing physicians in the U.S. is a graduate of a foreign medical school. The sixfold growth in their number over the past two decades results from national policy; but the place and conditions of practice have been controlled by state governments. The several states have used their jurisdiction over licensure in ways that have often been unfair and irrational. Recent trends toward uniform standards, however, may lead to more equitable assurance of professional competence, and to interstate mobility related to local and national needs.


[Lymphoid myelofibrosis or hairy cell leukemia]. By lymphoid myelofibrosis or hairy cell leukaemia or tricholeukaemia is meant an unusual haemopathic condition known only for the past few years. It is characterized pathognomonically by the presence of lymphocyte type cells with villous extroflexions, hence the name "hairy cell". Clinically the disease presents as an involutive myelopathy associated with splenomegaly, generally without any particular lymph gland involvement. The attention of students today is concentrated on the nature of the hairy cells; while some are inclined to admit their monocyte or histiocyte derivation, others consider that they derive from B lymphocytes. Therapeutically, almost everybody agrees that splenectomy is the only valid step. A case of H.C.L., which was typical from the clinical and laboratory viewpoints is reported. It is probable that certain haemopathic pictures once classified among atypical leucoses and lymphomas, would today be more correctly classed as hairy cell leukaemia.


Pelvic floor reconstruction following gynecologic exenterative surgery. At the end of exenterative procedures, the problem of bowel placement in the pelvic cavity denuded of peritoneum is still one of the most taxing and potentially troublesome problems of radical pelvic surgery. A technic which utilizes pelvic peritoneum to form a complete peritoneal diaphragm between the abdominal and pelvic cavities is described. Experience with this technic is with a small personal series of 15 patients which is presented.