Metastatic tumors in the temporal bone--a pathophysiologic study.

Nineteen temporal bones were examined from 11 patients who had metastatic temporal bone disease from a distant primary. The salient clinical features were: the high incidence of occult temporal bone involvement (7 of the 10 clinically documented cases), the considerable incidence of melanoma (3 of 10) and the variable correlation between clinical findings and pathologic localization of tumor in the temporal bone. Pathologic examination revealed two distinct modes of tumor spread within the temporal bone: 1) vascularosseous (petrous apex, mastoid, middle ear, external canal); and 2) perineural (nerves in IAC branches, labyrinthine endorgans). Every case was involved by one or both or these routes and no case of CSF-borne metastasis to the perilymphatic space was seen. The external canal was involved extensively in spite of an intact tympanic membrane. Since the presence of symptomatic or occult metastases in the temporal bone affects treatment and prognosis, they must be actively sought by the clinician.


Infections of the temporomandibular joint.

Three interesting case histories of infection of the temperomandibular joint are reported. An unsuccessful search of the recent literature was made in the hope that some assistance would be obtained in understanding the pathology. These infections must have been more common in the pre-antibiotic era and perhaps a search of the older literature would have been more fruitful.


Sexual impotence: the overlooked complication of a second renal transplant.

The incidence of sexual impotence in 20 men who have received at least 2 kidney transplants was 65% compared to 10% after the first transplant. The importance was transient (2 to 4 months) in both affected men after the first transplant but permanent (2 to 10 years) in 6 men after the second operation. We attribute this increased percentage of impotence to the second end-to-end arterial anastomosis that requires division of the internal iliac arteries. We suggest that the second transplant be placed end-to-side into the common iliac artery.


Vascular-induced erectile impotence in renal transplant recipients.

The incidence and pathogenesis of sexual dysfunction were studied in 16 male renal allograft recipients. In 9 patients with bilateral renal transplants in whom both internal iliac arteries were occluded nocturnal erections were absent and a penile arterial pulse could not be palpated. Of 7 patients with 1 renal allograft 4 demonstrated penile nocturnal tumescence and the penile pulse could be palpated in 5. Since interruption of pelvic arterial blood supply appears to contribute to impotence in renal transplant recipients consideration should be given to the use of the external iliac artery for arterial anastomosis. Two patients were treated successfully with an inflatable penile prosthesis.


Etiology, manifestations and therapy of acute epididymitis: prospective study of 50 cases.

There were 50 patients with acute epididymitis who were evaluated prospectively by history, examination and microbiologic studies, including cultures for aerobes, anaerobes, Neisseria gonorrhoeae, Chlamydia trachomatis and Ureaplasma urealyticum. Escherichia coli was the predominant pathogen isolated from the urine of men more than 35 years old, while Chlamydia trachomatis and Neisseria gonorrhoeae were the predominant pathogens isolated from the urethra of men less than 35 years old. The etiologic role of Escherichia coli and Chlamydia trachomatis was confirmed by isolation from epididymal aspirates from a high proportion of men with positive urine or urethral cultures for these agents. Chlamydia trachomatis epididymitis accounted for two-thirds of idiopathic epididymitis in young men and often was associated with oligospermia. Of 9 female sexual partners of men with Chlamydia trachomatis infection 6 had antibody to Chlamydia trachomatis, of whom 2 had positive cervical cultures for this organism and 2 others had non-gonococcal pelvic inflammatory disease. Antibiotic therapy with tetracycline was effective for the treatment of men with Chlamydia trachomatis epididymitis and should be offered to the female sex partners.


Operative blood loss in transurethral prostatectomy.

Operative blood loss was measured in 62 transurethral prostatectomies performed with a continuous irrigation suction resectoscope. Although blood loss per minute is comparable to levels in earlier studies the rapidity with which resection can be accomplished results in a marked reduction in operative blood loss.


Bacteriuria during closed urinary drainage: an evaluation of top-vented versus bag-vented systems.

For a 10-month period a top-vented closed urinary drainage system was evaluted and compared to a conventional bag-vented drainage system. The study involved random assignment of either system to all adult patients who required urinary catheterization. Criteria for exclusion included acute or chronic genitourinary tract infection and surgical interference with bladder function. The end point of the study was removal of any part of the system after at least 24 hours or a positive urine culture (greater than 10,000 colonies per ml. urine). The series included 236 patients. Distributions of patient age, sex and hospital service assignment (medical or surgical), antibiotic usage and duration of catheterization were similar in both study groups. Of 113 patients with the top-vented system 16 (14.2 per cent) acquired bacteriuria, whereas 13 of 123 (10.6 per cent) with the bag-drainage system acquired bacteriuria. No significant difference was noted. Antibiotics were used in 202 of 236 patients (86 per cent). Although usage delayed the appearance of bacteriuria in both groups isolates were more frequently yeasts and gram-negative organisms other than Escherichia coli.


The Denis-Browne hypospadias repair revisited.

Sixty-four cases of hypospadias repair by the Denis-Browne technique are reviewed. A urethrocutaneous fistula was the most common postoperative complication. Other less frequent complications were urethral stricture, residual chordee and improper location of the urethral meatus. These complications were corrected adequately and the results were satisfactory in all patients. This technique is simple, sound and trustworthy.


Adhesiveness to urinary tract epithelial cells of fecal and urinary Escherichia coli isolates from patients with symptomatic urinary tract infections or asymptomatic bacteriuria of varying duration.

Adhesiveness to human urinary tract epithelial cells was high for Escherichia coli strains isolated from patients with acute pyelonephritis and acute cystitis, and low for asymptomatic bacteriuria strains detected at screening. Escherichia coli bacteria causing asymptomatic reinfections, detected near the onset of bacteriuria, adhered more than those detected at screening. No difference in the adhesive ability was found between fecal isolates of the strain causing urinary tract infection, isolated at or before onset of bacteriuria, and the urinary strain in symptomatic or asymptomatic patients. Normal fecal Escherichia coli from non-bacteriuric patients adhered less than all other strains tested.


Parameters of gas urethral pressure profiles: Part I.

Carbon dioxide urethral pressure profiles were analyzed under the same conditions for 113 spina bifida and 87 non-spina bifida patients categorized by sex and age. Either duplicate or triplicate measurements were obtained from each patient for parameters of continence length, closure pressure and functional length. The degree of reproducibility of each parameter varied depending on age, sex and diagnosis. Except for 1 age-sex category closure pressure exhibited the smallest degree of relative variation for spina bifida patients. Results for non-spina bifida patients were mixed in that the measurement of no one parameter resulted in a uniformly smallest degree of relative variation. Reproducibility based on within patient variability generally was least for the continence length.


The use of percutaneous antegrade renal perfusion in kidney transplant recipients.

Percutaneous antegrade renal perfusion under ultrasound control may be performed safely in human renal transplant recipients with dilated pelviocaliceal systems. The study can be useful to define whether the presence of anatomic changes is accompanied by significant functional obstruction.


Upward migration of indwelling ureteral stents.

Polyethylene ureteral stents suspended from the kidney by a pigtail memory in the proximal end were placed bilaterally in a patient with ureteral obstruction secondary to retroperitoneal sarcoma. Both stents migrated upward and 1 migrated into the renal parenchyma. We recommend that these stents be inserted into the renal pelvis, as opposed to the calices, and that sufficient length be allowed to prohibit migration above the bladder.


Bacterial tracheitis.

During a 14-month period, eight infants and children were observed with an acute, infectious, upper airway obstructive disease with features common to both croup and epiglottitis. We have termed this distinct entity "bacterial tracheitis." All patients failed to respond to treatment for croup, including racemic epinephrine delivered by intermittent positive-pressure breathing. Direct laryngoscopy consistently revealed a normal epiglottis and aryepiglottic folds but marked subglottic mucosal edema. Tracheal suctioning yielded copius mucopus below the subglottic swelling. Gram stain of this material corroborated subsequent cultures: Staphylococcus aureus, six; group A Streptococcus, one; and Haemophilus influenzae (not typed), one. All patients required periodic tracheal suctioning for relief of upper airway obstruction. Six patients required endotracheal intubation; one required a tracheostomy. Bacterial tracheitis should be considered in the differential diagnosis of a young child with a croup-like illness that is refractory to conventional therapy.


Roentgenographic findings in PEEP therapy. Indicators of pulmonary complications.

We reviewed serial chest roentgenograms of 82 patients treated with positive end-expiratory pressure (PEEP) therapy for morbid pulmonary failure. No change was identified in the appearance of the chest roentgenograms in 49 patients. Thirty-three patients showed pulmonary hyperinflation, varying degrees of apparent clearing of alveolar or interstitial disease, and resorption of interstitial and pleural fluid. Lung hyperinflation and interstitial gas generally preceded pneumomediastinum and pneumothorax. These pulmonary complications were identified roentgenographically, often before they became clinically apparent. No consistent correlation was observed between these roentgenographic changes and the ventilatory pressures employed. The radiologist should be informed that a patient is receiving PEEP therapy, as the earliest signs of alveolar rupture--interstitial emphysema--are frequently subtle.


[Morphological studies on antibacterial activities of cefotiam (author's transl)].

1. Cefotiam was demonstrated to be more potent than cefazolin in its antibacterial activities against clinical isolates of E. coli, Klebsiella, Serratia, Proteus mirabilis, Proteus morganii and Proteus inconstans. MICs of cefotiam with 10(6) cells/ml inoculum size were considerably lower than those with 10(8) cells/ml. 2. Organisms lysed when exposed to cefotiam at concentrations higher than the MICs with 10(8) cells/ml. Morphological changes of organisms into filament occurred even at concentrations lower than the MICs with 10(6) cells/ml. This indicates that cefotiam is incorporated into organisms at remarkably low concentrations and exerts its antibacterial activities. 3. Cefotiam showed a high affinity for penicillin-binding proteins (PBP) 1A, 1Bs and 3. The formation of filament at low concentrations of cefotiam is possibly attributable to the high affinity of cefotiam for PBP 3 in addition to its high permeability through outer cell membrane. 4. As the antibacterial activities of cefotiam are displayed at lower concentrations, it is reasonable to consider that doses of cefotiam on clinical use can be reduced in comparison with those of conventional cephalosporins.


[Treatment of E. coli meningitis with cefmetazole. Report of two cases with favorable response and determination of the concentrations in CSF (author's transl)].

Two patients with purulent meningitis, of which causative organism was presumed to be E. coli, were treated with intravenous administration of cefmetazole, 300 approximately 400 mg/kg/day in 4 approximately 6 divided doses, and the following conclusions were obtained. 1) Clinical response was favorable and a complete cure was obtained without sequelae in both patients. There were no adverse reactions noted except for a mild and transient eosinophilia (12%) in one case. 2) Of two strains of E. coli recovered from CSF, one was sensitive to ampicillin, cefazolin and cefmetazole, among which cefmetazole had the highest bacterial activity. Although another strain was sensitive to cefmetazole, it showed resistance to cefazolin (greater than 12.5 microgram/ml) and to ampicillin (greater than 100 microgram/ml). 3) Concentrations of cefmetazole in CSF following 1 approximately 2 hours of its intravenous administration were either equal to or higher than those of ampicillin, which was given to the same patient for a short period of time. The concentrations in CSF were higher than 3.1 microgram/ml on each occasion except for in some specimens during the convalescent phase and exceeded well the MIC of the causative organism. 4) Based on the above results, cefmetazole is considered to be a potent antibiotic in the treatment of E. coli meningitis. Although further studies are needed as to the dosage, an intravenous administration at 4-hour interval appears to be warranted based on the studies that the half-life of the drug is short in CSF in animal experiments.


[Two-dimensional diffusion method for determining antimicrobial agents (author's transl)].

Intending to improve the accuracy of determination of antibiotics, a two-dimensional diffusion method using large agar plate was introduced. Three antimicrobial agents, ampicillin, PC-904, and tobramycin, were used. Inhibition zones of B. subtilis on the agar plate were measured which were formed as a result of diffusion of these agents. The relationship between the concentration of antimicrobials and the size of inhibition zones was studied. Plotting the data-points on the graph, it was predicted that there might be a relationship of quadratic equation between the diameter of inhibition zone and the logarithm of concentration of the agents. On the other hand, mathematical considerations were taken to find out a physical principle or an equation which governs the diffusion of antibiotics in the agar. Assuming that antibiotics spreads in the agar after the principle of simple diffusion, an equation was lead which shows how the antibiotics distributes in the agar in relation to time. The equation was written as, (formula: see text) where, S is amount of antibiotics, D diffusion constant, r distance from the center of diffusion, t period during which the diffusion proceeds. As a result of the mathematical calculations mentioned, it was confirmed that the relation between the size of zones and the logarithm of concentration of antibiotics is described by a quadratic equation as predicted on the basis of experimental data.


Mutation of human cells by kerosene soot.

The polycyclic aromatic hydrocarbon fraction of a kerosene soot induced forward mutation in human diploid lymphoblasts when coincubated with Sprague-Dawley rat liver postmitochondrial supernatant. Two components of the kerosene soot extract, benzo[a]pyrene (BP) and cyclopenta[cd]pyrene (CP), were also tested. TP was not mutagenic at the concentration found in the soot extract, although it was active at higher concentrations. The amount of CP present could account for approximately 8% of the total mutation observed with the soot. The results were compared to data obtained previously in a similar mutation assay in Salmonella typhimurium. The protocol described permits the facile assay of mutation at the hgprt locus in human lymphoblasts; such mutation is induced by compounds of complex mixtures requiring mixed-function oxygenase activity for metabolism to genetically active derivatives.


Selection of a low antigenic variant subline from the TA3St ascites carcinoma by repeated passage of antibody-coated cells through anti-immunoglobulin columns.

TA3St mouse ascites carcinoma cells of strain A origin (H-2a) were coated with anti-H-2a serum and passaged through mouse immunoglobulin-anti-immunoglobulin columns repeatedly prior to each in vivo passage. After six passages in vivo, a variant cell line emerged with significantly decreased H-2a expression and an increased ability to grow progressively across H-2 allograft barriers. Chromosome analysis of the variant cell line revealed no major changes in karyotype. The application of the method for studies on the generation of immunoresistant tumor cell variants and on the relationship between antibody-binding sites and rejection target sites is discussed.


Effect of adriamycin and Corynebacterium parvum in tumor-bearing mice: modulation of response to sheep red blood cells.

Administration of adriamycin and Corynebacterium parvum alone in C57BL/6J mice bearing Lewis lung carcinoma stimulated the direct (19S) and indirect (7S) plaque-forming cell (PFC) response specific for sheep red blood cells. Thus adriamycin appears to possess some immunostimulatory effect on tumor-bearing mice with much less effect than C. parvum alone. Simultaneous administration of adriamycin and C. parvum decreased the PFC response compared to that for C. parvum alone. This decrease may indicate that drug-vaccine interaction could produce some inhibition of the strong immunostimulatory effect of C. parvum as measured by the PFC response in tumor-bearing mice. The immunostimulatory effect of adriamycin and C. parvum administered as a single agent or in combination was associated with significant splenomegaly. The results of this study could be helpful in clinical situations when these agents are used alone or combined.


Metabolism of 1,3-bis(tetrahydro-2-furanyl)-5-fluorouracil in mice.

1,3-Bis(tetrahydro-2-furanyl)-5-fluorouracil (FD-1) is a new masked compound of fluorinated pyrimidine and a derivative of 1-(tetrahydro-2-furanyl)-5-fluorouracil (FT-207). The pharmacokinetics of FD-1 and FT-207 were compared in the livers and kidneys of control mice and in mice (DD males) pretreated with phenobarbital. The half-time in the liver of FD-1 orally administered was about 40 minutes, whereas that of FT-207 in the liver was about 3 hours. Hepatic concentrations of 5-fluorouracil (FUra) originating from FD-1 were three to five times as much as those originating from FT-207. FD-1 formed 3-(tetrahydro-2-furanyl)-5-fluorouracil (3-T-F-FU) and FT-207 by a ratio of about 3 to 10, respectively. Although FD-1 had a short period of half reduction, FT-207 and 3-T-F-FU had a half-time lasting for 3--4 hours. In contrast, the renal concentration of FD-1 was one-third that of FT-207. Oral administration of FD-1 to the mice pretreated with phenobarbital elevated the FT-207 and 3-T-F-FU levels in the livers to twice the levels in the control mice and further elevated the FUra levels in the livers to twice the levels in the controls. These results indicate that FD-1 is catabolized in the liver by microsomal enzymes (including cytochrome P450) faster than is FT-207, which consequently enlarges the hepatic pool of the intermediates on the way to FUra formation.


The mother as incubator-after delivery.

Decreases in body temperature of 10 nude babies who were held next to their mothers' bodies were compared with the decreases in temperature of 9 babies who were kept in heated beds. After 15 minutes both groups of babies had identical temperatures of 98.1 degrees F, which is well above the acceptable minimum of 96.9 degrees F. It appears that mothers who desire to hold their babies immediately after delivery can be permitted to do so without concern by the staff that the babies will become chilled.


Localization of urinary procoagulant in the human kidney.

By the indirect immunofluorescent and immunoenzymatic techniques with monospecific antiserum against urinary procoagulant (a tissue factor which accelerates blood coagulation), we found the urinary procoagulant in the kidney distributed to the loop of Henle and distal convoluted tubules. In these areas urinary procoagulant was found in association with the luminal and intercellular borders as well as in the cytoplasm of epithelial cells. Both the descending and ascending limbs of Henle were equally stained. The cytoplasmic staining was patchy in distribution among cells of distal tubules and was predominantly localized in the supranuclear areas. Glomeruli, the proximal tubular cells, the vascular wall, and the interstitium were not stained. There was, however, fluorescent staining along the epithelial layers of the Bowman's capsule, which was observed only in the frozen sections. Casts in the distal tubules were also positively stained. These findings suggest that urinary procoagulant is synthesized in the epithelial cells of these particular parts of nephron and is secreted into urine, although its physiologic roles and pathologic significance are not entirely known.


[Falk's technic of extranasal dacryocystorhinostomy in cases of tear duct stenoses (author's transl)].

The lacrimal sac is totally sacrificed and rebuilt into an accessory nasal sinus using the extranasal dacryocystorhinostomy in the treatment of nasolacrimal duct stenosis as described by Falk in 1961. Using this method, 62 patients with tear duct stenoses were operated in our clinic since 1959. According to etiology, the stenoses were in 5 cases connatal, in 17 traumatic, in 32 inflammatory and in 8 iatrogenic. In only 4 cases (6,4%) failures were reported. - the operative indication has at present been expanded to include stenosis of the lower tear ducts (presaccal stenosis) or in cases where the lacrimal sac has been obliterated (saccal stenosis). In these cases the stenosis is dissected and a silicon tube (introduced via the tear point into the inferior lacrimal duct and drained into the nose) is left in position.


[Massive air embolism complicating artificial ventilation (author's transl)].

A 1740 g premature infant being treated by intermittent positive pressure ventilation with PEEP because of respiratory distress syndrome died from massive air embolism occuring in two attacks on the 14th and 16th day of life. The pathogenesis of systemic air embolism in neonates with respiratory distress syndrome is discussed and the difficulties in diagnosis as well as the importance of this complication regarding possible cerebral damage are considered. There is no conclusive information about the incidence of this serious event available at present.


[Circulating immune complexes in bronchogenic carcinoma: relation to extent of disease and to therapy (author's transl)].

In sera of 72 patients with lung cancer, 20 patients with various benign lung diseases and 34 age matched controls circulating immune complexes were determined by column chromatography on Sepharose 6 B and subsequent testing of the eluate for macromolecular IgG as well as by inhibition of radiolabelled C1q binding to sensitized sheep erythrocytes. Whereas in both control and benign lung disease-sera complexes could be detected in less than 5%, sera of lung cancer patients showed macromolecular IgG in 83% and C1q reactive material in 53% at the time of diagnosis. Patients with metastases exhibited a significantly higher percentage of positive reactions than those without metastases (macromolecular IgG 93%/68%, C1q 71%/28%). The size of the complexes increased with the extent of disease. So far, no signficiant changes in circulating immune complexes could be demonstrated id pretherapeutic values were compared with those after X-ray-, chemo- or immunotherapy with one exception, which is an increase of C1q reactive material after radiotherapy.


Effects of duration of daily illumination on reproductive organs and fertility of the meadow vole (Microtus pennsylvanicus).

The objective of this study was to determine the optimum simple, constant photoperiod for voles in a laboratory colony. Voles from an established colony maintained at 22 degrees C with a photoperiod of 14:10 hours of light:dark were transferred at 50 days of age to a photoperiod of 12:12, 14:10, 16:8 or 18:6 hours of light:dark. From Day 96 to Day 102, the 9--10 females per treatment group were paired with a male. Body and gonadal weights, spermatogenesis, ovarian activity and pregnancy were evaluated on Day 110. Reproductive function and body weight of both male and female voles maintained on a photoperiod of 16:8 exceeded (p less than 0.05) values for voles exposed to 12:12 hours of light:dark and tended to be more favorable than for voles in the 14:10 and 18:6 groups.


Perfusion fixation of the newborn opossum: equipment and techniques.

A 30-gauge cannula was used to perfuse fixative through the fragile heart of a newborn opossum. The cannula was gently maneuvered into the heart and held in place with a specially designed manipulator. The flow rate of the fixative was regulated with an infusion set connected to the cannula.


Tamm-Horsfall glycoprotein: ultrastructural immunoperoxidase localization in rat kidney.

Tamm-Horsfall urinary glycoprotein (TH) is the primary constituent of urinary casts. The intracellular distribution of TH in normal rat kidney was determined by immunoelectron microscopy using horseradish peroxidase-labeled antibodies and compared with morphologic localization of TH by immunofluorescence and light immunoperoxidase microscopy. Electron microscopy revealed an intracellular localization of TH restricted to the thick ascending limb of the loop of Henle (ALH). In this nephron segment, TH was distributed on and between adjacent intercellular membranes and infolding intracellular membranes at the base of these cells, within Golgi vacuoles, apical vesicles, and on the luminal membranes. Macula densa cells were negative, although typical ALH cells across the lumen of the same tubular segment were positive. Other renal segments were negative for intracellular TH. The unique distribution of TH is consistent with the known function of the ALH as the diluting segment of the nephron. We speculate that the aggregation and gel formation of TH on and between ALH surface membranes may restrict water movement across the ALH. This influence on permeability would be an important role for TH in the generation of concentration gradients for the countercurrent multiplier system of the kidney.


The infected pacemaker pocket.

Between January, 1963, and September, 1978, a total of 1,789 pacemakers were implanted at Henry Ford Hospital. Infection at the site of implantation developed in 19 instances for an incidence of 1.06 percent. The most common organism cultured was Staphylococcus epidermidis, and conservative treatment was successful with these patients. In all patients with organisms other than Staphylococcus epidermidis, reimplantation of a new unit in a new, clean site was required.


Cancer of the breast. Mammography and thermography.

Quality mammography with knowledgeable interpretation is now a widely utilized and reliable procedure. It permits clinically occult lesions to be detected and clinically obvious or indeterminate lesions to be managed more intelligently. Abnormal mammographic signs are well defined, as are their differential diagnoses. Thermography of the breast is a younger science than mammography and must mature before its full potential can be realized. It is clear that thermography today cannot be considered an adequate prescreening technique to obviate further examination, as was once anticipated. Used in conjunction with physical examination and mammography it can serve to reinforce suspicions, and the high acceptability of the examination is an impetus to further clarify its role in detection.


Cancer of the breast. Epidemiology.

Though the etioolgy of mammary cancer in humans remains obscure, epidemiologic data, animal models and in-depth studies of patient populations are providing insights. The rapid progress and sophistication of these investigations promise to clarify with increasing precision the genetic and environmental determinants of risk, if not the fundamental cause. The importance of this information cannot be overestimated; inherent in it lies the possibility of prevention.


[Value of the LH-RH test in secondary hypothalamic amenorrhea].

Forty-three patients with secondary hypothalamic amenorrhea and 17 healthy women in the early and late folliculin phases of the normal cycle were tested with 100 micrograms synthetic LH-RH i. m. The LH and FSH concentration in the plasma was measured before and after the injection of synthetic LH-RH. The patients were divided into two groups, according to the degree of estrogen deficit. The basal concentration and maximum LH increase in the group of hypothalamic amenorrhea with a milder estrogen deficit proved to be significantly higher than in the group with a higher estrogen deficit (22.2 +/- 7.2 versus 12.7 +/- 9,3 p less than 0.01 and, respectively, 74.8 +/- 17.00 versus 34.4 +/- 11.2, p less than 0.01). In healthy women with the normal cycle, the basal LH and FSH concentrations and the maximum increase after the LH-RH application were significantly higher in the late than in the early folliculin phase. The basal LH and FSH concentrations and their increase after the LH-RH injection appear to depend on the concentration of endogenous estrogens, both in normal cycle and in secondary hypothalamic amenorrhea.


Cancer of the breast. Staging methods, primary treatment options and end results.

A totally satisfying concept of treatment is not easy to formulate from the complex and often conflicting results of local therapeutic interventions for breast cancer. It seems evident that clinically occult cancer is often beyond the pale of both resection and irradiation at primary treatment, particularly when cancer is found in regional lymph nodes. Despite all combinations of local treatment, the ultimate risk of failure correlates more closely with the stage of the disease at the time of treatment than with the particular form of treatment. Thus the extent of disease must be considered the major, perhaps the ultimate determinant of prognosis. Because, under controlled conditions, several therapeutic alternatives have appeared to provide virtually identical end results in terms of survival and ultimate dissemination of the disease, the adequacy of control within the field of treatment may, in fact, be the most meaningful end result of local treatment. The experience that has accumulated with treatment of breast cancer supports the thesis that removal of the breast accomplishes all that can be achieved in terms of curing the disease, and wider treatment with surgery or irradiation serves only to improve the prospects for local control. Halsted demonstrated this principle with his radical mastectomy and it still seems to be the case. This fact provides further impetus for detecting and treating cancer while it is still localized to the breast. With these generalizations in mind some empirical observations can be added. An anatomic fact is that multiple microscopic foci of cancer that are not evident clinically are often present in the mammary parenchyma. Undisturbed, at least some, and perhaps eventually all, of these foci of cancer progress to become clinical cancers. Thorough removal of the entire breast (the entire mammary parenchyma) eliminates this particular hazard and, one may presume, terminates the disease if it is still limited to the breast. Removal of the underlying pectoralis major muscle provides additional margin around the tissues primarily involved, but sacrific of the muscle is apparently needless unless it is directly invaded by cancer. Microscopic metastases are also often present in regional lymph nodes without being clinically detectable and, left untreated, have the capacity to enlarge and become clinically apparent. Routine wide removal of regional lymph nodes improves the control of cancer at these sites when metastases are present, but whether it improves the chances for cure is doubtful. The fact is that approximately 25 per cent of patients with axillary metastases enjoy prolonged survival free of recurrence, some remaining well even after thirty years (Adair et al., 1974). Whether they would survive as well without removal of the metastases is uncertain. Desease-free survival is highest if metastases are removed while still microscopic, but this phenomenon may simply reflect treatment at an earlier phase in the evolution of the disease...


Cancer of the breast. Surgical management.

We have reviewed a variety of surgical techniques for mastectomy, endocrine gland ablation and the management of pathologic fractures and pleural effusion. With their judicious use for the chronic disease of mammary cancer, many persons can be cured and many others can be given long periods of symptom-free survival. With (1) attention to proper indications and contraindications for surgery, (2) surgical detail and (3) good patient care, very few patients will be made worse.


Cancer of the breast. Endocrine and hormonal therapy.

Additive hormonal therapy remains the treatment of choice for disseminated breast cancer in postmenopausal women. Patients with hormone-dependent tumors receive excellent and long-lasting palliation from alterations in the hormonal milieu. Now that hormone receptor assays are clinically available, responses can be accuratedly predicted in a large percentage of cases. Tables 11--6 is a summary of additive hormonal therapy in postmenopausal patients. Endocrine ablative therapy remains of primary importance in premenopausal women because of the superior results, but androgens or antiestrogens may be helpful when patients are not surgical candidates. Castration continues to be the initial approach, with adrenalectomy or hypophysectomy reserved for promising candidates. In postmenopausal women the initial choice is estrogens. The exceptions are those patients with metastases limited to bone, when androgens excel because of an equivalent objective response and superior subjective and metabolic effects. Patients who respond to estrogens and then progress are observed for a rebound regression following the discontinuation of estrogen therapy. Whereas some who do not respond to androgens will respond to estrogens, the converse does not appear to be true (Kennedy, 1974). Currently progestins are the secondary hormonal agent of choice in postmenopausal women, but they may be displaced by antiestrogens as more data become available. In general, if a patient's tumor lacks estrogen receptors or the patient fails to respond to an adequate trial of endocrine or hormonal therapy, one should proceed directly to cytotoxic chemotherapy. A suggested plan for the integration of endocrine with hormonal therapy and both with other forms of palliation is diagrammed at the end of Chapter 12.


Mammary carcinoma and pregnancy.

Among premenopausal women approximately 7 per cent of newly diagnosed breast cancers are accompanied by pregnancy, with most patients being in the fourth decade of life. The prognosis of these patients, and of those whose cancers are diagnosed soon after pregnancy, is generally less favorable than that of nonpregnant females, but if age and stage of disease are comparable, pregnancy per se has little influence upon prognosis. Mastectomy is as effective for pregnant patients as for others and the chance of spontaneous abortion is small. Therapeutic abortion does not improve the chances for cure of patients with clinically localized cancer. Effective endocrine therapy or chemotherapy for advanced or disseminated breast cancer does require therapeutic abortion, and an early pregnancy is best terminated without delay. For pregnancies near term the decision depends greatly upon the desire of the patient for a child. Unless therapeutic needs are urgent, intervention can often be delayed temporarily without significant deterioration of the patient. Pregnancies subsequent to a mastectomy have little bearing upon continued well-being, and as long as the patient is clinically free of cancer no therapeutic benefit can be expected from interrupting them. A decision for future pregnancies should be individualized with due regard for the risk of recurrence and the desirability of completing one's family while still reasonably young. Progress with the treatment of breast cancer depends in part upon the appreciation that cancers do occur during pregnancy and lactation, that they are best diagnosed early and that they are curable. Pregnancy should neither deter a prompt diagnosis nor delay definitive treatment.


Cancer of the male breast.

Cancer of the male breast is infrequent but not rare. In contrast to women, men are generally older at its onset and present with more advanced cancers. In comparable circumstances, they are probably equally curable with radical surgery. The tumor is sensitive to irradiation and often responds to endocrine ablations, making the latter exceptionally useful for palliation of systemic metastases. Hormones and chemotherapy can also be beneficial.


Age associated increase of single-stranded regions in the DNA of mouse brain and liver cells.

The touch smears of brain cells and hepatocytes of young and senescent mice were stained with antibody to cytidine nucleoside by an indirect immunofluorescence technique and subsequently combined with fluorescence cresyl violet staining of DNA. Nuclear binding of the antibody which reacts only with denatured or single-stranded regions in the DNA was seen only in the tissues of an aging animal. No such DNA lesion was detected in the epithelial cells of the gastrointestinal tract at any age. This type of DNA alteration is supposed to accumulate in the slowly renewing and non-replenishing tissues as a function of aging. The antibody was found not to react with the cells in S phase as demonstrated by 3H-thymidine autoradiography on a smear of newborn hepatocytes after the double fluorescence staining with cresyl violet and anti-cytidine antibody.


Progress in cytogerontology.

The finite in vitro lifetime of cultured normal cells is interpreted to be aging at the cellular level. In addition to the inverse relationship between donor age and population doubling potential (PDP), a number of biochemical and physiological increments and decrements occur prior to the cessation of cell division. The reconstruction of replicating normal human cells from the nuclei of "young" cells and the cytoplasm of "old" cells (and the reverse) suggests that the nucleus governs PDP. Several morphological changes were found to occur in late phase III cells held for up to one year in culture. Autoradiography studies show that (1) a cell population may be composed of several subpopulations, each of which is at a different stage in its life history and (2) lipid synthesis is affected much less as cells age than is DNA, RNA and protein synthesis. Changes occurring in the genetic program of individual cells seem to be the most tenable hypothesis to explain fundamental causes of aging.


The pathogenesis of atherosclerosis.

The pathogenesis of atherosclerosis is hypothesized to occur as a response to various forms of injury to the lining arterial endothelial cells. The resulting endothelial alterations could potentially lead to interactions between platelets in the circulation and the underlying subendothelial connective tissue or with the altered endothelial cells themselves. Such interactions provide an opportunity for platelet degranulation and release of a platelet-derived growth factor. This factor has been shown in cell culture to be an extremely potent mitogen and will induce DNA synthesis and cell multiplication of a number of cells including smooth muscle cells, fibroblasts, and other mesenchymally derived cells. Chronic endothelial injury and repeated interactions between platelet-derived mitogens, plasma components, and the underlying arterial smooth muslce cells would promote the progression of the intimal proliferative lesions of atherosclerosis that lead to the clinical sequelae associated with this disease process.


Hemopoietic stem cell heterogeneity: use of cell cycle-specific drugs to look for age-associated alterations.

Hemopoietic tissue is vulnerable to perturbations, and data show that it is an appropriate tissue in which to look for age-associated alterations. This tissue has a high regenerative capacity, is composed of a heterogeneous population of stem cells that are capable of self renewal or differentiation, or both, and is sustained by a pool of resting cells. The heterogeneity of bone marrow has made characterization of the cellular elements difficult. Techniques commonly used to identify and quantify the various maturation levels of hemopoietic stem cells and the limitations of these techniques are discussed. Most techniques used to assay age-associated changes in bone marrow have not differentiated between specific cellular alterations or shifts in the distribution of the cellular elements. In particular, it has been difficult to determine the stability of the non-dividing stem cell because of the low incidence of this cell (6 per 1000) and the lack of a specific assay for this important cell type. The use of cell cycle-specific drugs has provided quantitative information on specific subpopulations of hemopoietic stem cells and seems to be the most promising approach towards determining qualitative and quantitative differences in the hemopoietic stem cells of young and old individuals.


Interrelations of dietary and hormonal effects in aging.

A model system is proposed which suggests that aging involves a genetically programmed process encoded within specific regions of the central nervous system and is characterized by a gradual ascendency of inhibitory over excitatory influences. The resulting neural suppression reduces tonic stimulation to the hypothalamus-pituitary-endocrine axis, the autonomic nervous system and the spinal cord, and promotes the secretion of inactive pituitary hormone, the involution of the somatic musculature (smooth, cardiac and striated) and the compromise of secretomotor function. Experimental observation from nutritional restriction studies which include the prolongation of the life-span and delays in reproductive and homeostatic aging, degenerative coat changes and the appearance of neoplasia, are considered from the perspective of this hypothesis.


Glial cells: modulators of neuronal environment.

Studies of glial cells in neural tissue culture systems suggest that glial cells subserve different functions during development and aging of the central nervous system and that they may help modulate the neuronal environment by virtue of their responsiveness to hormones and other intrinsic factors. There is a marked proliferation of glial cells during early stages of brain development, probably reflecting the involvement of glial cells in myelination and other growth processes. Studies in culture suggest that proliferation of glial cells can be induced by steroid hormones. The migration rate of glial cells from cerebellar explants of embryonic chick brain grown in organotypic culture was measured in control and hormone-treated explants. Treatment with cortisol, corticosterone, estradiol, and progesterone significantly elevated glial cell migration from the tissue explants. The influence of steroid hormones on glial cells may be mediated via a steroid intracellular mechanism. In C-6 glioma cells and in chick embryo dissociated brain cell cultures consisting predominantly of glial cells, 3H-corticosterone was shown to accumulate by a saturable but non-specific retention mechanism. In contrast, the accumulation of 3H-corticosterone by predominantly neuronal cultures was both saturable and specific. Glial cells in culture exhibit certain age-related changes, including changes in resting membrane potentials and in cellular responses to hormone treatment, as measured by changes in incorporation of 3H-leucine into protein and incorporation of 3H-uridine into RNA. The possibility that glial cells in vivo may likewise exhibit differential responses to hormones throughout the lifespan suggests that hormones may markedly influence cellular aging.


Individual and institutional variables which may serve as indicators of quality of medical care.

This article is a critical review of empiric studies, in the medical care literature of the past two decades, that investigated associations between characteristics of physicians and medical care institutions and some measure of the quality of medical care given by them. The intention is to identify those characteristics of physicians and medical care institutions which can be considered the best indicators of the quality of performance to be expected, given the present state of knowledge. The analysis discusses 18 such characteristics but derives a list of 14 which appear to be the best choice of indicators on which further research might focus. It would be possible to design a survey instrument based on these characteristics, which, if upheld by empiric testing, could serve as a crude assessment tool for third parties needing to make quality comparisons between medical care institutions.


[Spontaneous carotid-cavernous fistula: angiogammagraphic findings and computer analysis (author's transl)].

The case of a 74-year-old woman with a carotid-cavernous fistula is reported. She was admitted to hospital with oliguria and generalized edema. After improvement of the edematous condition she suddenly presented intense, pulsatile, frontal cephalea. It was more intense on the right side and was accompanied by nausea and vomiting and swelling of the right eye with reddening, sharp pains, difficulty of movement, and loss of vision. Cerebral angiogammagraphy was practiced in anterior-posterior and right lateral view. A righ carotid-cavernous fistula was discovered with drainage through the superior ophthalmic vein and the deep venous system. The angiogammagraphic findings can be considered characteristic; it is important to obtain activity/time curves from various areas of interest in order to evaluate the degree of shunting and the pathway of drainage from the fistula. There was a good coorelation with the arteriographic findings. Eight cases of carotid-cavernous fistulas diagnosed with radionuclids have been published previously, though the activity/time curves were not determined by computer for any of them.


[Acute effects of bumetanide in healthy subjects. A comparative study with furosemide (author's transl)].

A comparative test on the efficacy of bumetanide and furosemide was made in ten healthy individuals. The effect on diuresis and urinary excretion of ions and metabolites after the acute oral administration of 1 mg bumetanide and 40 mg furosemide was observed. From the results of the experiment it can be concluded that the effectiveness of bumetanide is about 15 percent superior to that of furosemide with a later but more sustained effect. The excretion of Na and K was similar with both compounds but the chloruresis was more intense with furosemide. Although the excretion of uric acid was increased too, it was proportionally less than the increase of the diuresis obtained with both drugs. A noticeable increase of the urinary excretion of calcium caused by bumetanide stands out. The authors suggest its possible therapeutic usefulness in patients with hypercalcemia as well as its possible mechanism of action on the reabsorption of calcium. In every case tolerance of both drugs was good and there were no signs of toxicity or adverse side effects. Generally speaking the doses of 1 mg bumetanide and 40 mg furosemide were judged to be equally powerful.


[Determination of endotoxemia by means of the Limulus test in patients with septicemia (author's transl)].

The Limulus test was applied to determine bacterial endotoxemia in 42 patients suspected of having septicemia. Hemocultures were practiced simultaneously. A comparative study is made of the two presently most commonly recommended tests for neutralizing the factors inhibiting the endotoxin-amebocyte reaction. According to our results the dilution of plasma in isotonic saline solution is preferable; we obtained a sensitivity of determination of 0.0001 micrograms of E. coli endotoxin per ml of plasm. Seventeen cases of bacteriemia were diagnosed, only one of which with sepsis due to Serratia marcescens responded positively to the limulus test. The test was positive in three of the remaining 25 patients. Two of them had liver failure. The basic role that liver failure plays in the high level of endotoxin in the blood is discussed. The prognostic value of this test and the low correlation between bacteremia and endotoxemia are also commented on


[Infrared contact coagulation. Clinical application in liver and spleen (author's transl)].

Infrared-contact-coagulation is a new principle for hemostasis of bleeding parenchyma and was used in 33 patients with 41 bleeding surfaces, 22 in the liver, 19 in the spleen. Hemostasis was achieved in all but three deep lacerations without further complications. Accidental bleeding, biopsy, and resection were the main indications; biopsy and partial resection of the spleen was achieved.


[First experiences with voice-rehabilitative laryngectomy, as described by Staffieri and Amatsu (author's transl)].

Report on six cases of tracheo-esophageal shunt-operations as described by Staffieri, and four cases operated according to Amatsu. Both methods hold in common the construction of a slit-like anastomosis between trachea and hypopharynx, respectively upper esophagus. According to Staffieri the anastomosis is made towards the hypopharynx, according to Amatsu towards the upper esophagus. Advantages of the Amatsu-method, as compared to the Staffieri-method, are as follows: 1. no additional hypopharynx-mucosa is required to establish the shunt. Therefore, it is also suited for tumors with loss of pharynx-mucosa, 2. when constructing the anastomosis, mucosa of esophagus, and of trachea, is directly and seamlessly connected so that no granulations or polyps can develop, 3. voice and speech is learned earlier, 4. less aspiration. Both methods have the following disadvantages: 1. for voice-production, the tracheostoma has to be occluded with a finger, 2. a certain amount of aspiration must always be reckoned with, though, up to now, no patient asked for an occlusion of the fistula. At the time being, we are recommending the shunt-method to those patients only who expressly ask for it because of the possibility of early voice-production, and a better quality of speech. Or we recommend it to such patients as would otherwise not consent to laryngectomy.


A computer-aided system for the diagnosis of hepato-biliary diseases. A comparison with the performance of physicians.

A computer-aided system has been developed for the diagnosis of disease of the liver and biliary system. The program is based on the use of 30 indicants, all of which are available within six hours after patient's admission: 18 of them are clinical signs, 12 are laboratory parameters including routine liver-function tests. To date, the program concerns 52 hepato-biliary diseases. From the analysis of the 30 items collected in any patient, the diagnoses are computed according to Bayes' theorem and printed by decreasing order of probability. The performance of the program was tested using records of patients with fully proven diagnosis. The first diagnosis given by the computer was correct in 57 per cent of the cases. In 80 per cent, the right diagnosis was among the first four proposed. When the performance of the model was compared to that of physicians, the number of correct answers was roughly the same for the computer and for the specialists in hepatology; in contrast, the computer's responses were far better than those of general practitioners. These results demonstrate the efficiency of the program for the diagnosis of hepato-bilitary diseases and its potential interest for helping clinicians in decision-making.


Computers and medical language.

This review article presents principles and problems of automated processing of medical language data. Work on automated processing of information in medical language is surveyed. References and a bibliography are provided as an introduction to the field.


Unusual uses of the deltopectoral flap.

For the past decade the deltopectoral flap has become the premier reconstructive tool in oncologic head and neck surgery. It has achieved a level of reliability that has not been approached by any other single flap. Its most common application has been replacement of lost skin either from planned surgical resection or necrosis following ablative surgery. This paper illustrates a few unusual clinical settings in which the deltopectoral flap was used instead of other conventional methods: 1. Use of the same delopectoral flap in two different locations. 2. Employment of the base of the deltopectoral tube for skin coverage. 3. Successful application of bilateral simultaneous deltopectoral flaps. 4. Three-stage reconstruction of the cervical esophagus in a tenuous metabolic setting. 5. Two-stage technique for soft palate replacement. 6. An alteration of the design of the conventional deltopectoral flap because of an upper pectoral abscess.


[Laser indicator of immunological reactions].

The instrument described was designed for measurement of the scattered light integral intensity; the light is scattered within a volume where immunologic reaction takes place. Operation speed and sensitivity are the merits of this instrument, as compared to standard ways of the immunologic reactions observation.


[Attachment for stereoroentgenogrammetric photography].

Construction and operation principles of an automated stereoradiographic set and a stereoradiocomparator with a portable digital computer are described. This technique, when used for radiotopometric purposes, allows rapid determination of precise quantitative data in clinical conditions about internal structure of organism.


[Value of indirect immunofluorescence test in the diagnosis and prognosis of intestinal and extra-intestinal amoebiasis in endemic areas (author's transl)].

A comparative study of 108 patients with patient amoebiasis and a reference group of 85 free of the disease but living in endemic area. The positivity of the test has been more frequent and has given higher titers in the extra-intestinal forms than in the intestinal ones. The positivity rate in the reference group has been 15 per 100.


[Post-therapeutic evolution of patients presenting an immunological suspicion of trypanosomiasis without parasitological proof in support of it (author's transl)].

The authors have recently preconised systematic treatment of every human being who is fluorescent positive. The evolution of antibodies of 26 subjects parasitologically negative and positive only in FAT test is described here after treatment with Arsobal. Statistically the antibodies decrease progressively like antibodies of Sleeping Sickness patients of first period usually did. Therefore it is possible to conclude that immunological suspects who were treated were realy sick and rightly cured.


The genetic map of transfer RNA genes of yeast mitochondria: correction and extension.

Ninety five rho- mitochondrial DNA's of Saccharomyces cerevisiae were compared for their deletion structure by means of 15 genetic markers and 22 tRNA genes. The patterns of co-deletion and co-retention of different tRNA genes allowed us to determine their positions with respect to each other. The deduced order of tRNA genes was consistent with the order of the genetic markers established by independent genetic approaches. Our previously proposed mitochondrial tRNA gene map has been revised and extended. Transfer RNA genes, corresponding to all 20 aminoacids, and two isoacceptor tRNA genes were localized. The possible position of each tRNA gene has been indicated on the physical map of mitochondrial DNA. Seventeen tRNA genes are carried by a narrow region representing less than 20% of the wild type genome.


Isolation and properties of a plasmid which expresses the E. coli Su+7 amber suppressor tRNA gene.

The gene of the amber suppressor tRNA derived from tRNATry, Su+7, has been inserted into a col E1-derived vehicle by selecting for its expression. Despite selection for a suppressor phenotype, and the plasmid's stable presence at ca. 180 copies cell during balanced growth, the level mature tRNA maintained by the gene is less than that of the normal haploid tRNATry locus in the bacterial chromosome. Transfer RNA genes, both the plasmid Su+7 gene and chromosomal tRNA's are expressed during inhibition of protein synthesis. During, e.g. chloramphenicol inhibition, Su-7 and Su+7 tRNA can be elevated similarly in the plasmid-containing cell; Su+7 reaches levels of molecules/cell which ordinarily characterize a major tRNA. The recombinant plasmid, but not the cloning vehicle alone, has a more general effect on tRNA levels; accumulation of tRNA from three chromosomal tRNA loci including tRNATry, continues during extensive isoleucine limitation. The plasmid therefore contains a locus which probably alters the relaxed-stringent circuit, whose effect is disseminated to at least 3 widely separated loci.


The structure of the DNA containing the E. coli tRNATry gene.

Using the pMB9 recombinant plasmid pMY3, which contains a functional gene for the tRNATry mutant Su+7, the EcoRI fragment containing the tRNATry gene is mapped and oriented with respect to the HindIII site in the tetracycline region of pMB9. Complete HpaII and HaeIII maps of the EcoRI fragment are derived. The Su+7 tRNA gene is placed by hybridization to these fragments, and the tRNA gene is oriented by using the restriction sites for HinfI, TaqI, and HpaII in the tRNA gene itself. A tRNAAsp gene is shown to lie adjacent to tRNATry, and is also placed and oriented in the map. The RI fragment itself originates in a locus adjacent to, and transcribed in the same direction as, the ribosomal RNA genes of phi 80d3. The implications of the structure of the cloned DNA for its previously measured regulatory and tRNA gene activities are discussed. In particular, the effect on the regulation of RNA synthesis is attributable to an E. coli DNA sequence, but cannot be due to the presence of a normal tRNA promoter on the plasmid.


Relaxation of stable RNA synthesis by a plasmid-borne locus.

The plasmid pMY3, which was constructed so as to express the Su+7 amber suppressor tRNA gene, also relaxes control of stable RNA synthesis in stringent cells. The relaxation is not growth medium or strain-dependent and does not occur in the presence of the vehicle alone. When expression of the effective sequence is diminished, in a lysogen of phi 80d3 ilv+Su+7, the sequence no longer affects RNA synthesis. The relaxation is general, extending to all or almost all tRNA loci, including tRNAs located in the ribosomal spacer regions, and to all ribosomal RNAs. Relaxed plasmid-carrying strains are still able to elevate guanosine tetra- and penta-phosphate levels in response to amino acid starvation, but steady state levels are somewhat diminished. Aminoacyl-tRNA falls to control levels when the plasmid-carrying strain is deprived of amino acid. Therefore, the relaxed strain perceives amino acid starvation, but does not respond normally. These properties define a novel locus which relaxes stringent control.


Studies of a plasmid coding for tetracycline resistance and hydrogen sulfide production incompatible with the prophage P1.

The plasmid pIP231, determining tetracycline resistance and hydrogen sulfide production is shown to belong to incompatibility group Y and to code for a restriction and modification system. Unlike the IncY plasmids, P7 and P15B, plasmid pIP231 shows only little genetic and physical homology with P1 prophage.


Nuclear mutations in Saccharomyces cerevisiae which increase the spontaneous mutation frequency in mitochondrial DNA.

Fourteen mutants have been identified in which the frequency of spontaneous mutations in mitochondrial DNA is increased. As well as increasing the frequency of mutations to resistance to erythromycin, oligomycin and spiramycin, all the mutants also show changes in the frequency of spontaneous petite induction. None of the mutants has any effect on the frequency of spontaneous nuclear mutations. Nine of the mutants are in one complementation group and five are in another. The phenotype of both groups is caused by a single nuclear mutation.


Genetic analysis of an osmotic sensitive Saccharomyces cerevisiae mutant.

The genetic analysis of VY1160 sorbitol dependent, osmotic sensitive yeast mutant led to the identification of three different nuclear recessive mutations. Two of them, designated sorb- and ts1 are closely linked to one another. The mutation sorb- determines the lysis, while the mutation ts1 increases the ability for lysis of the sorbitol dependent cells. The third mutation ts2 segregates independently from the other two and confers the sensitivity of VY1160 mutant cells towards rifampicin.


Alterations of lipid metabolism in mice injected with endotoxin.

Some alterations in lipid metabolism in mice were observed by the intraperitoneal injection of endotoxin from Salmonella typhimurium. The content of serum triglyceride increased markedly in poisoned mice 16-24 hr postintoxication. The level of free fatty acid (FFA) in the serum of endotoxin-administered mice decreased in inverse proportion to an increase in the injected dose of endotoxin. The electrophoretic analysis of the serum lipoprotein on cellulose acetate membrane showed that pre beta-lipoprotein increased markedly and that FFA fraction in the poisoned mice sera disappeared 18 hr postintoxication. The activity of hormone-sensitive lipase in adipose tissue was elevated appreciably 2 hr after injection, but decreased more significantly after 18 hr than that in fasted control mice. On the other hand, the activity of lipoprotein lipase decreased in the post-heparin serum and adipose tissue 3 hr postintoxication, and decreased significantly after 16 hr. There were no significant differences between changes in the formation of active glycerol (alpha-GP) and in the activity of alpha glycerophosphate dehydrogenase (alpha-GPDH) in the mice liver with or without administration of endotoxin, and after 16 hr levels of both hepatic alpha-GP content and alpha-GPDH activity in poisoned mice showed a tendency to be slightly lower than those in fasted control mice.


Microsomal aspects of carcinogenesis and neoplasia.

Neither immunologic nor genetic concepts of carcinogenesis have yet been decisively confirmed, and epigenetic theories, as formulated so far, are either non-predictive or insufficiently consistent with morphologic and experimental evidence. Computing data, concerned with carcinogenic mechanisms and neoplastic changes at the level of the endoplasmic reticulum, may lead to a new coherent understanding of tumor pathogenesis. Carcinogenic agents initiate biophysical perturbations, chemical alterations and conformational transitions in the membrane lattice of the endoplasmic reticulum. Foremost among the resulting neoplastic changes is an increased, irreversible separation of polyribosomes from membranes of the ergastoplasm. The carcinogenic process, apparently, deletes a protein required for polysome attachment. Since microsomal cytochromes can be synthesized by membrane-bound polysomes only, the translation of genetic information for their biosynthesis is irreversibly restricted. A similar, self-perpetuating deficiency may be postulated for the polysome attachment protein. Activities, depending on cytochromes P-450 and b5, are hampered, e.g. those of the monoxygenase system. Cholesterogenesis is derepressed. Ratios of phospholipids/cholesterol are decreased, and lipid-protein complexes, altered both in structure and function. Another distinct effect of the membrane-polysome separation is the unmasking of thiol-disulfide exchange enzymes which, in turn, stimulate the biosyntehsis of proteins and of deoxyribonucleotides involved in cell replication.


Ovarian function and the immune system.

A hypothesis is presented on the interaction between the immune system and ovary in the regulation of the reproductive system and in the origin of some of its disorders. It has been suggested that the beginning, duration and age dependent failure of ovarian ovulatory function depends among other things on the adequate relationship between the immune system and approriate ovarian target structures. The cyclicity of ovarian function is considered to be primarily dependent on the induction of a specific cyclic immune response to the ovary. Similarly, the selection of a species-specific number of ovulating follicles during sexual maturity is thought to be ensured by immune mechanisms. This hypothesis, on the role of the immune system in regulation of ovulatory ovarian function respects the physiological effect of gonadotropins and steroids on the ovarian structures. The interaction between the ovary and the hypothalamus-pituitary system appears to be modulated by the relationship between the ovary and the immune system.


The possible role of hypertension in aggravating hemolytic episodes in G-6PD deficient persons.

Approximately 13 percent of American Negro males carry a mutant [A-variant] glucose-6-phosphate dehydrogenase enzyme in their red blood cells that predisposes them to hemolytic episodes following exposure to oxidant drugs such as primaquine. Most hemolytic episodes to standard prophylactic treatment are mild and self-limited, but as many as 2 percent of Negro males develop severe hemolysis when similarly treated. The exaggerated response may be due in part to the combination of G-6-PD deficiency and hypertension since hypertension can cause red cell fragmentation, and the stressed cells of G-6-PD deficient person would be more sensitive to such fragmentation.


[Contribution to the clinicopharmacological fundamentals of the orthostasis syndrome (author's transl)].

We have carried out therapeutic trials in 20 orthostatic patients (ages ranging between 20 and 35 years) with unequivocal signs of a static-labile circulatory disturbance. The test programme was used three days per week over a three-week period (medication of Amphodyn-retard). The test programme consisted of measurements of the systolic and diastolic blood pressure, the pulse pressure and the pulse rate at one-minute intervals during a resting, loading (standing) and recovery phase, each covering seven minutes; further measurements of the flicker fusion threshold (FFT) in each case in the 3rd and 4th minute in each of the three study phases. Compensation of disturbances of orthostatic regulation was observed as a result of the treatment: Unchanged systolic and diastolic blood pressure during loading, narrowing of the pulse pressure did not occur, and the pulse rate increased only slightly. There was an increase in the flicker fusion threshold as an expression of the stabilization which was slightly significant. The associated subjective symptoms improved.


Computerized system for the measurement of gallstone volume, in vivo.

Instrumentation and methodology for the determination of gallstone volume from standard roentgenographic views have been developed. Evaluation of size is made by using video viewing, an operator-set electronic window, intercept registers, and the programming of a PDP/8E for handling the data. The programming makes corrections for geometric factors arising from anatomical variations, corrections for variations in technique, roentgenographic magnification, and provides for the use of several roentgenographic views. The precision of the measuring (video) technique is approximately 0.5 mm as measured on the roentgenogram. Estimates and test data indicate that this system should be capable of an absolute accuracy within 10% of the true volume.


Gene activation during immune reaction.

In cell-free systems the addition of antigen stimulates the synthesis of informational RNA (i-RNA) which exhibits the following properties: It codes for the entire antibody molecule, it codes for the synthesis of regulator protein which initiates transcription of i-RNA with the correspondent informational content from DNA, it is a template for an an i-RNA dependent RNA polymerase, it is a template for an i-RNA dependent reverse transcriptase. The i-RNA may exist in a state of latency in cells. The product of reverse transcription of i-RNA is i-DNA which can be used to transcribe further i-RNA of the same specificity. Similar to i-DNA is an extracellular DNA which codes also for antibody and from which i-RNA can be transcribed. The data presented are summarized in a scheme of the flow of information during immunological reactions. It could be shown that there exist three different types of extrachromosomally synthesized molecules--i-RNA, i-DNA and extracellular DNA--which bear immunological specific information. These extrachromosomal states of information may be relevant for the generation of antibody diversity.


Isolation and binding properties of leucyl-tRNA synthetase from Escherichia coli MRE 600.

A procedure for the large-scale isolation of leucyl-tRNA synthetase from E. cole MRE 600 is described: The enzyme was purified about 320-fold to homogeneity by precipitation with cetyl-trimethyl-ammonium bromide, two consecutive chromatographies on DEAE-cellulose and three on hydroxyapatite with an over-all yield of 4%. The molecular weight of leucyl-tRNA synthetase from E. coli MRE 600 was found to be 99 000 daltons. Bindings studies by ultracentrifugation and equilibrium partition showed that the enzyme binds leucine, leucyl-adenylate and tRNA Leu, each in a 1 : 1 stoichiometry. For ATP only a very weak binding to the enzyme could be observed, which did not allow the evaluation of the complex stoichiometry. The presence of ATP was not required for the binding of leucine or tRNA to leucyl-tRNA synthetase from E. coli MRE 600.


Messenger ribonucleoproteins (informosomes) and RNA-binding proteins.

Messenger ribonucleoproteins, first discovered in 1964 in our laboratory as free mRNA-containing particles of fish embryo cytoplasm and designated as informosomes, proved to have a universal occurrence in eukaryotic cells. Messenger ribonucleoproteins of different intracellular localization such as free cytoplasmic non-translatable informosomes, translatable messenger ribonucleoproteins in polyribosomes and nuclear pre-mRNA-containing particles are characterized by a number of features common for all of them. However, the transport from the nucleus into the cytoplasm as well as the transition from the free non-translatable state into the polyribosome-bound translatable state are accompanied by essential changes in the protein moiety of the particles. The existence of free RNA-binding proteins in eukaryotic cells has also been shown. These proteins seem to represent a pool for the formation of messenger ribonucleoproteins (informosomes). It has recently been demonstrated that the eukaryotic translation factors and, in particular, both the elongation factors and some initiation factors are among the cytoplasmic RNA-binding proteins. It is suggested that the mRNA in eukaryotic cells at different stages of its life time carries on itself the proteins which are required for its own biogenesis, processing and transport (nuclear informosomes), for its existence in a temporarily inactive state (free cytoplasmic informosomes) and for its functioning as a template (polyribosomal informosomes): omnia mea mecum porto.


On pre-messenger RNA and transcriptions. A review.

From the present review integrating old and new data emerge a few principles of gene expression in eukaryotes, and an infinite variety of possible mechanistic details generating the overal pattern. The few principles, most of which are not fundamentally new, may thus be summarized. 1) The eukaryotic genome is subdivided into transcriptional units: into transcriptons which are subject to individual activation controlled at DNA level. 2) Viral genomes may contain one or a few transcriptons, while cells of multicellular organisms contain from 3 x 10(3) in diptera up to an estimated 2 x 10(5) in birds and mammals. 3) Transcriptons may include one or several coding sequences. 4) Transcriptons vary considerably in size: in mammals and birds their size spectrum falls into the 2,000 to 20,000 bp range. 5) Units of coding information constituting one message (genes) and, possibly, units of regulative information are frequently broken up and stored within the transcripton in sub-genic blocks (of so far unknown significance) in general located at a certain distance from the 5' and 3' transcript terminals which are determined by the promotor and terminator signals. 6) The gene, in its specific definition as the functional unit underlying the phenotype, is in general constituted posttranscriptionally by the processing mechanisms from the mosaic of its genomic subunits in the transcripton; segments of coding, service and regulative sequences are recombined within themselves and with each other, polygenic transcripts separate into their unit messages. 7) Activated transcriptons produce pre-mRNA; these primary transcripts are colinear with the DNA of the transcriptional unit. 8) Primary pre-mRNA is processed into secondary pre-mRNA's by extragenic cleavage and intragenic ("splicing") processing, giving rise stepwise to functional mRNA. During this process chemical modifications as methylation, 5'-terminal capping and 3'-terminal polyadenylation take place. 9) Translation yields either potentially functional polypeptides or polycistronic polyproteins subject to further processing. 10) Processing is a regulated process; it involves many of the possible phases and mechanisms of post-transcriptional regulation (cf. 39, 40).


[Immunogenic DNA of Shigella sonnei bacteriophage].

Immunogenic and some other properties of Sh, sonnei "Ufa" phage DNA were investigated. This DNA has been shown to possess high immunogenic activity indicating the presence of an unusual base. The base is distinct from 5-OmC, glucosylated 5-OmC, 5-OmU or dihydroxypentyluracil. The phage DNA is double stranded and has the molecular weight of 1.11.10(8) daltons and Tm=85.5 degrees C. Base sequences common with T4 DNA have been found using the competition DNA-DNA hybridization technique. Experiments on cross neutralization of intact phage with the corresponding antisera confirm the existence of common antigenic determinants in T4 and Sh, sonnei "Ufa" phage particles.


[Substrate specificity of E. coli glutamate decarboxylase].

Interaction of highly purified E. coli glutamate decarboxylase with a number substrate analogs was studied. Decarboxylation of the following amino acids was demonstrated: gamma-methylene glutamate, threo-beta-hydroxyglutamate, allo-gamma-hydroxyglutamate, threo-beta-methylglutamate, homocysteate, aminoadipate and cysteinesulfinate. The Km and either Ki or I50 values were determined for these compounds. The final products of the interaction of glutamate decarboxylase with these analogs have the same absorption spectra and capacity for reactivation by pyridoxal-P, as has the pyridoxamine-P form of the enzyme. Thus, decarboxylation of all the amino acids, mentioned above, was probably associated with the side reaction of transamination to coenzyme in the active center. Binding of aliphatic dicarboxylic acids or of valeric acid by glutamate decarboxylase leads to a slight shift of absorption spectra and of circular dichroism spectra from 420 to 423--425 nm. The following compounds fail to be bound and decarboxylated by the enzyme: gamma-aminobutyrate, D-glutamate, L-glutamine, 3,3-dimethylglutarate, methioninesulfone, methioninesulfoxide, norvaline, gamma-hydroxy-gamma-methylglutamate, erytro-beta-methylglutamate and erythro-beta-hydroxyglutamate.


[Methylation of E. coli RNA polymerase with dimethylsulfate].

DNA dependent RNA polymerase from E. coli was methylated with dimethylsulfate. After the methylation the enzymatic activity was lost. Addition of two methyl groups per enzyme monomer completely inactivated enzyme with respect to RNA synthesis but couldn't prevent enzyme binding to DNA. Methylated enzyme was able to form tight complexes with DNA and to compete with the native enzyme for the formation of rifampicin resistant complex with DNA. The ratio of the binding constants of the native and methylated enzymes to DNA was determined to be equal to 3. Methylated enzyme was not able to form the first phosphodiester bound as revealed from pyrophosphate exchange reaction studies.


[Conformational transition of DNA within the B-family as revealed by anisotropy circular dichroism].

The native T2 DNA in solutions with different LiCl concentrations was oriented by pumping through a capillary device similar to that described in rf. 1. The change of the parallel, deltaepsilon parallel, and perpendicular, 2 delta epsilon perpendicular, components of circular dichroism tensor was studied as a function of salt concentration. Positive (delta epsilon parallel 280) and negative (2 delta epsilon perpendicular 280) components were shown to increase in absolute magnitude, so that there was a monotonous drop in the longwave band magnitude of the isotropic delta epsilon 280 as a result of compensation; the shortwave band magnitude of the isotopic delta 245 and its anisotropic components were constant. It is proposed that while the helix is winding as a result of LiCl rise progressive tilting of base pairs takes place. It was also shown that the glucosylated T2 DNA in solution is wound to a greater extent than the non-glucosylated DNA of the same GC-content.


[Hydroxyapatite thin-layer chromatography of nucleic acid].

The present work is concerned with a sensitive and fast micromethod for separation of single- and double-stranded molecules of nucleic acid by hydroxyapatite (HAP) thin-layer chromatography. The thin layers were obtained by precipitation of ground HAP particules into the surface of the plates in water. Chromatography in sodium phosphate buffer makes it possible to separate from 1 to 50 micrograms of nucleic acids for 30--50 sec. Thereby double-stranded molecules remain at the starting line, whereas single-stranded DNA or RNA follow up the solvent. For quantitative assay of nucleic acids by HAP thin-layer chromatography, the plates were scanned in UV light, radioactivity was measured without extracting substances from HAP and DNA and RNA were eluted with the help of phosphate buffer. A simple and accurate determination method has been suggested consisting in dissolving HAP in perchloric acid followed by hydrolysis of nucleic acids and spectrophotometry of solutions. The retrieval of the material after chromatography in 99 +/- 2%, the mean determinations error is 2--3%. The conditions are described for extraction, after thin-layer chromatography, of desalted and concentrated DNA, ready for use in later experiments. The paper describes a method: for determination of the degree of DNA nativity; quantitative determination of DNA in solutions, containing admixtures; separation of synthesized RNA from its precursors and from the DNA template; assay of DNA thermostability; investigation of the kinetics of DNA reassociation and DNA-DNA hybridization. Some results obtained from hydroxyapatite thin-layer chromatography are discussed.


[Induction and repair of breaks in DNA chains in vivo due to the imbalance between DNA and protein synthesis].

A sudden induction of the imbalance between the rates of DNA and protein synthesis in the cell (by nalidixic acid or by thymine starvation) results in the stabilization of breaks in DNA chains in vivo. Such "imbalance induced breaks" represent gaps in DNA chains formed with the participation of exonuclease V. Stabilization of the "imbalance induced breaks" is accompanied by DNA degradation and cell death. Restoration of the imbalance between the rates of DNA and protein syntheses by balanced inhibition completely prevents the stabilization of breaks in cells with competent repair systems. Balanced inhibition of intracellular DNA and protein synthesis decreases the rate of repair and permits to see the sequence of induction (stabilization) and disappearance (repair) of breaks in DNA chains in vivo. Repair of breaks occuring on the background of balanced inhition of DNA and protein synthesis decreases the extent of DNA degradation and completely prevents death of E. coli cells with completely functional repair systems.


[Compact structure of the small E. coli ribosomal subunit and its RNA studied by fluorescence spectroscopy and sedimentation analysis].

Analysis of the temperature dependence of fluorescence polarization of ethidium bromide adsorbed on the double helical fragments of 16S RNA's hairpin loops was used to characterize the intramolecular flexibility of RNA in the free state and within 30S subunit. We show that the local mobility of RNA segments is strongly limited by the tertiary structure of 16S RNA and ribosomal proteins reinforce these limitations. It was suggested that the mechanism of the temperature dependent RI particle activation involved the temporary increase of the local mobility of RNA segments in RNP which favored the formation of the new intraribosomal contacts. A comparison of sw 20 dependences of RNA and 30S subunit on Mg+ and K+ concentrations leads to the proposal that RNA in the small subunit in the physiological conditions has the stressed conformation. This conformation is maintained by the Mg2+-dependent RNA-RNA interactions induced by ribosomal proteins and specific only for the subunit.


[Addition of the fluorescent label to the 3'-OH end of DNA and the 3'-OH end of nascent RNA].

3'(2')-O-acyl derivatives of the uridine triphosphate were synthesized. Acyl residues contained fluorescent dye; fluoresceine or rodamine C. Optical properties and stability of UTP analogues were studied. Their ability to serve as the substrates for calf thymus terminal deoxyribonucleotidyl transferase and E. coli RNA polymerase was also examined. It was shown that both enzymes were able to use tested analogues as substrates. Incorporation of the analogues into nascent RNA and DNA chains inhibited the synthetic reaction because of primer inactivation. The rate of the incorporation of the analogues showed an exponential time dependence


[Repression of the enzyme inducible syntheses in Escherichia coli K12 mutant with a deleted ptsH gene].

The genome of lambda phage with thermosensitive repressor was integrated into the pts region of the E. coli chromosome. Such a lysogenic culture behaves as a pts mutant at 30 degrees. Heating of cells of this strain leads to the induction of lambda prophage and formation of deletions in the pts region. A mutant with a deletion covering ptsH gene was isolated after prophage induction. The deletion nature of pts mutation was confirmed in genetic and biochemical experiments. It was shown that the deletion is small and does not involve ptsI and lig genes. The isolated deltaptsH mutant possesses all characteristics of pts mutants: pleiotropic impairment of transport and utilization of a number of carbohydrates, repression of the enzyme inducible synthesis and resistance to catabolite repression with glucose. These data (together with earlier ones) allow us to conclude that the phosphorylated form of HPr is involved (in direct of indirect manner/ in activation of DNA transcription.


[Influence of protein bound with single-stranded DNA on the synthesis of RNA and poly(A). III. Protein product of F1 gene 5].

Influence of the protein product of F1 phage gene 5 (protein 5) on the synthesis RNA and poly(A) in vitro was studied. It has been shown, that protein 5 has no effect on the transcription of the native DNA by E. coli RNA-polymerase, but completely prevents RNA and poly(A) synthesis on the denatured or single-stranded DNA at the protein/DNA ratio 10:1. Protein 5 inhibits poly(A) synthesis with oligo(dT)9 and oligo(dT)12 as a template, preventing binding of enzyme to the oligonucleotide. After the initiation of the poly(A) synthesis the inhibition becomes considerably wearer. The biological function of the inhibition of the transcription by "unwinding" proteins is discussed.


[Fragment reaction catalyzed by E. coli ribosomes].

It has been shown that 50S subunits of E. coli MRE-600 ribosomes catalyze the reaction of N-(formyl)-methionyl ester of adenosine 5'-phosphate acting as peptide donor, with Phe-tRNA or CACCA-Phe serving as a peptide acceptor. The reaction is stimulated by cytidine 5'phosphate and inhibited by lincomycin, puromycin and chloramphenicol. The obtained results show that the structure of the donor site of peptidyltransferase is completely assembled on the 50S subunit and 30S subunit is not required for its formation.


[Repair of the interstrand cross links in DNA: role of restriction system "K"].

The interstrand cross links formed in the DNA after psoralenplus light treatment of bacteria E. coli K12 and bacteriophage lambda are repaired by the restriction and modification system "K." Strain E. coli C, which does not have the restriction and modification system is 2 times more sensitive to psoralen plus light treatment, than the wild type E. coli K12. But these strains are equally sensitive to inactivation by UV-light (254 nm). By studing the sedimentation of bacterial DNA in alkaline sucrose it was found that cells of E. coli C are deficient in filling of gaps, appearing in parental DNA after enzymatic excision of the "arms" of cross links. In experiments with bacteriophage lambda (W-reactivation, prophage-reactivation), treated by psoralen plus light, it was shown that E. coli C and mutants of E. coli K12 rk-mk+/- are deficient in non-recombination lexA-dependent repair of cross links. The function of restriction and recognition of restriction and modification system are essential for repair of cross links. It is suggested that the restriction and modification system "K" acts either as a regulatory protein or as a protector of single-stranded regions of DNA against nuclease attack.


[Unprimed synthesis of poly (d(A-T)), catalyzed by a preparation of Escherichia coli DNA polymerase I].

The initial events of the de novo synthesis of poly[d(A-T)], catalyzed by preparations of E. coli DNA-polymerase I, were investigated. The data provide evidence that deoxynucleoside diphosphate: oligonucleotide deoxynucleotidyl transferase (dNDP-transferase), the enzyme which is able to catalyze unprimed polymerization of dNDP, participates in the process of initiation. This conclusion is based on the following data: 1) preincubation of E. coli DNA-polymerase I preparation with dADP and dTDT abolishes a lag-period in the poly[d(A-T)] synthesis; 2) dithiothreitol and N-ethylmaleinide, inhibitors of dNDP-transferase, inhibit de novo synthesis of [d(A-T)]-copolymer by preparations of E. coli DNA-polymerase I but do not effect primed synthesis ensured by this enzyme. High concentration of the substrate have similar effect. Using two-dimentional thin-layer chromatography and microcolumn chromatography on TEAE-cellulose we have shown that preliminary incubation of DNA-polymerase I preparations with dADP and dTDP results in the synthesis of short oligonucleotides (from di- to decanucleotides). Hydrolysis of these oligonucleotides with dilute sulfuric acid demonstrates that among the reaction products prevail oligoadenylates and oligothymidylates, but an appreciable amounts of heterooligomers including oligo[d(A-T)] were revealed as well. The model of so called de novo synthesis of regular polynucleotides is proposed, according to which dNDP-transferase, an accompanying enzyme in the preparations of DNA-polymerase I E. coli, is carrying out the synthesis of short oligonucleotides which form template-primer complexes repeatedly replicated by the DNA-polymerase I E. coli.


[Isolation of free cytoplasmic informosomes].

The review concerns methods used by different authors for isolation of free cytoplasmic informosomes. The results of centrifugation in sucrose gradient and in density gradients, electrophoresis, ion-exchange, gel-filtration and affinity methods are described. It is concluded that these methods do not give good purification of informosomes from ribosomes and free proteins. Urgent necessity of a more careful control of the purity of informosomes isolated is emphasized.


[Influence of ionic strength on RNA-polymerase structure].

Chromatography of RNA polymerase holoenzyme preincubated under different ionic strength conditions on the DNA agarose column was studied. Ratio of two peaks identified to be core and holoenzyme was analysed. In the range of 0.15 to 0.05 M KCl the relative content of the holoenzyme peak gradually decreased from 100 to 50%. At the same time a peak of free sigma-subunit appeared as detected by the chromatography on DNA agarose gel A-1.5 m. The dissociation of half of the sigma-subunit amount occured within the enzyme dimer-monomer transition range. The results suggest that the dimerization follows the equation: E sigma + E sigma in equilibrium with E2 sigma. Reconstitution of the RNA polymerase holoenzyme from purified core enzyme and sigma-subunit was also studied by the same method. Reconstitution did not occur at a low ionic strength (0--0.1 M KCl), but takes place at ionic strength of 0.2 M or higher. Possible function of the dimerisation of the enzyme in search of promoter site and regulation of RNA synthesis is discussed.


[Lambdoid phage structural proteins and antigens].

The composition of structural proteins of lambdoid phages such as lambda, phi 80 434 divided by molecular weights was determined by means of SDS-disc-electrophoresis in a 15% polyacrylamide gel. The proteins of the same phages were divided by isoelectric points using an isoelectric focusing in a 5,25% polyacrylamide gel with 8 M urea and a gradient pH 7.0--3.5. The both methods brought out a composition character of the virion proteins and illustrated the high degree of similarity among the structural proteins of phages lambda and 434 and a far less similarity among the proteins lambda and phi 80. The antigenic composition of the lambdoid phage was determined and the basic antigenes were identified on one-dimensional and two-dimensional immunoelectrophoregrams. The appreciable immunochemical affinity of basic antigenes of the lambda and 434, but a partial affinity of the phages lambda and phi 80 were found. The basic protein of the head pE proved to be immunochemically similar for all three phages.


[Effect of single-stranded and double-stranded breaks on the melting temperature of phage T2 DNA].

The effect of single- and double-stranded breaks in DNA phage T2, on the melting temperature of this DNA in the 0,05 M SSC solution, was investigated. The number of cleavages per 1000 nucleotide pairs varied in the range of 0 to 10. It is shown that single- and double-stranded breaks affect the melting temperature with approximately (within 20%) the same efficiency. The relationship between the melting temperature shift (delta Tm) and the number of cleavages is non-linear. The magnitude of the effect is characterized by delta Tm of 2 +/- 0.4 degrees C for the average inter-cleavage distance of 200 base pairs. It is shown that the observed melting curves are non-equilibrium ones, which is probably due to the fact that the effect of cleavages on the melting temperature is largely results from the complete and practically irreversible separation of strands.


[Present state of antiviral therapy (author's transl)].

Application of antiviral drugs in viral infections is still limited to few diseases. Most of the preparations available induce strong side effects since these substances not only interfere with virus replication but also with cell metabolism. Therefore, only in severe diseases such as herpes simplex encephalitis or complications of varizella zoster infection such a treatment is advisable. From a clinical point of view an antiviral drug would be desirable which selectively inhibitis virus growths without affecting the host. This can already be achieved with interferon. However, at the present time the quantities of interferon needed for such treatment are not available.


[Role of RNA-polymerase in gene activity regulation of E. coli RNA-polymerase mutants with a pleiotropic effect. I. Physiological and biochemical studies].

Four Rifr-mutants of E. coli B/r (rpo B401, rpo B402, rpo B403, rpo B409) which differ from the wild strain in one or more phenotypic properties besides rifampicin resistance were obtained. Transfer of the mutant Rifr-alleles into the parent strain gives the latter all the properties of the mutant. This indicates that the new properties are due to the pleiotropic effect of Rifr-mutations. Biochemical studies of the properties of RNA-polymerases from the mutants and the parent showed that some new properties of the mutants could not be explained by the appearance of analogous properties in the mutant RNA-polymerase itself. They seem to be caused by alteration in functional activity of the mutant enzyme, particulary, alteration of its control properties during transcription. The function of the beta-subunit in genetic transcription is discussed.


The effects of the ultraviolet-protecting plasmids pKM101 and R205 on DNA polymerase I activity in Escherichia coli K-12.

The mutagenesis- and repair-enhancing plasmids pKM101 and R205 were introduced into a series of Esherichia coli K-12 polA mutants including two temperature-sensitive mutants. Polymerase levels in extracts of these strains were assayed using an activated DNA template. In none of the cases did the presence of the plasmid in the strains change either the initial rate of incorporation of [3H]thymidine triphosphate into acid-soluble material or the subsequent degradation of the template at longer reaction times. Neither did the presence of the plasmids affect the proportion of N-ethylmaleimide-sensitive polymerase activity detected. Previous studies have reported increased polymerase I-like activity of polA mutants of Salmonella typhimurium and Pseudomonas aeruginosa upon introduction of mutagenesis- and repair-enhancing plasmids. Our experiments indicate that, at least, such an increase in polymerase-I-like activity is not an obligatory phenotype associated with these plasmids.


[Heat denaturation of paramyosin].

Heat denaturation of paramyosin and its two proteolytic fragments has been studied by scanning microcalorimetry. It has been shown that paramyosin denaturation proceeds in several stages, each stage corresponding to the melting of a cooperative block in the macromolecule. The junctions between the blocks are the most strongly affected by proteolytic fragments. Enthalpies of paramyosin and its fragment denaturation and their temperature dependencies have been measured. It has been shown that the mean value of the specific enthalpy extrapolated to 110 degrees is noticeably lower than for globular proteins and is close to the values obtained earlier for L-meromyosin and tropomyosin. A conclusion has been drawn that the lower, in comparison with globular proteins, value of the denaturation enthalpy of alpha-superhelical structure is of a general character.


The mutagenicities of safrole, estragole, eugenol, trans-anethole, and some of their known or possible metabolites for Salmonella typhimurium mutants.

Safrole, estragole, anethole, and eugenol and some of their known or possible metabolites were tested for mutagenic activity for S. typhimurium TA1535, TA100, and TA98. Highly purified 1'-hydroxyestragole and 1'-hydroxysafrole were mutagenic (approximately 15 and 10 revertants/micromole, respectively) for strain TA100 in the absence of fortified liver microsomes; trans-anethole and estragole appeared to have very weak activity. 3'-Hydroxyanethole was too toxic for an adequate test. Supplementation with NADPH-fortified rat-liver microsomes and cytosol converted 3'-hydroxyanethole to a mutagen(s) and increased the mutagenic activities for strain TA100 of 1'-hydroxyestragole, 1'-hydroxysafrole, estragole, and anethole. No mutagenicity was detected for safrole or eugenol with or without added NADPH-fortified liver preparations. The electrophilic 2',3'-oxides of safrole, 1'-hydroxysafrole, 1'-acetoxysafrole, 1'-oxosafrole, estragole, 1'-hydroxyestragole, and eugenol showed dose-dependent mutagenic activities for strain TA1535 in the absence of fortified liver microsomes. These mutagenic activities ranged from about 330 revertants/micromole for 1'-oxosafrole-2',3'-oxide to about 7000 revertants/micromole for safrole-2',3'-oxide. The arylalkenes, their hydroxylated derivatives, or their epoxides did not show mutagenic activity for strain TA98, except for 1'-oxosafrole-2',3'-oxide, which had weak activity. Since the arylalkenes are hydroxylated and/or epoxidized by hepatic microsomes, hydroxy and epoxide derivatives appear to be proximate and ultimate mutagenic metabolites, respectively, of the arylalkenes.


[Quantitative studies of interaction of polyuridylic acid with 30S subunits of ribosomes of Escherichia coli].

Polyuridilic acid of average molecular weight 18 000 binds to the 30S subunits with stoichiometry 1 : 1 but two kinds of 30S.poly(U) complexes with different stability are formed. The main reason for such heterogeneity was found to be due to the presence or absence of ribosomal protein Sl in 30S subunits. In its presence the association constant of 30S.poly(U) complex is equal 2.7.10(8) M-1, and in the opposite case it is much less 1.5.10(6) M-1. In the same conditions (20 mM MgCl2, 200 mM HN4Cl, 0 degrees) the association constant of binary complex Sl.poly(U) is equal 5.10(7) M-1.