Colorimetric analysis with N,N-dimethyl-p-phenylenediamine of the uptake of intravenously injected horseradish peroxidase by various tissues of the rat

1. A method is described for the colorimetric determination of peroxidase with N,N-dimethyl-p-phenylenediamine. The amount of red pigment formed by peroxidase is proportional to the concentration of enzyme and to the time of incubation during the first 40 to 90 seconds. The influence of the concentration of enzyme, N,N-dimethyl-p-phenylenediamine, H(2)O(2), the time of incubation, pH, the temperature, and the possible interference by oxidizing and reducing agents of tissues has been tested. 2. The method has been used to follow the uptake of intravenously injected horseradish peroxidase by 18 different tissues of the rat over a period of 30 hours. The highest concentration of the injected tracer enzyme was found in extracts of kidney, liver, bone marrow, thymus, and spleen. Considerable amounts were taken up by pancreas, prostate, epididymis, and small intestine. Lower concentrations were found in extracts of lung, stomach, heart, and skeletal muscle, aorta, skin, and connective tissue. No uptake was observed by brain and peripheral nerve tissue. 3. Tissue homogenates containing high concentrations of the injected peroxidase, in general also showed high or average activity of acid phosphatase. 4. Six hours after intravenous administration, the liver contained 27 per cent, the kidney 12 per cent, and the spleen, 1.4 per cent of the injected dose. 5. Approximately 20 per cent of the injected peroxidase was excreted in the urine during the first 6 hours, and the concentration of peroxidase in blood serum and urine fell exponentially during this time. After 6 hours, only low concentrations were excreted in the urine but low enzyme activity was still detectable after 30 hours. Approximately 6 per cent of the injected dose was excreted in the feces from 6 to 20 hours after administration. 6. After feeding through a stomach tube, low concentrations of peroxidase were found in blood serum and urine. Considerable variations in the extent of absorption from the gastrointestinal tract were observed in individual rats.     

Colorimetric Analysis with N,N-Dimethyl-p-phenylenediamine of the Uptake of Intravenously Injected Horseradish Peroxidase by Various Tissues of the Rat

1. A method is described for the colorimetric determination of peroxidase with N,N-dimethyl-p-phenylenediamine. The amount of red pigment formed by peroxidase is proportional to the concentration of enzyme and to the time of incubation during the first 40 to 90 seconds. The influence of the concentration of enzyme, N,N-dimethyl-p-phenylenediamine, H₂O₂, the time of incubation, pH, the temperature, and the possible interference by oxidizing and reducing agents of tissues has been tested. 2. The method has been used to follow the uptake of intravenously injected horseradish peroxidase by 18 different tissues of the rat over a period of 30 hours. The highest concentration of the injected tracer enzyme was found in extracts of kidney, liver, bone marrow, thymus, and spleen. Considerable amounts were taken up by pancreas, prostate, epididymis, and small intestine. Lower concentrations were found in extracts of lung, stomach, heart, and skeletal muscle, aorta, skin, and connective tissue. No uptake was observed by brain and peripheral nerve tissue. 3. Tissue homogenates containing high concentrations of the injected peroxidase, in general also showed high or average activity of acid phosphatase. 4. Six hours after intravenous administration, the liver contained 27 per cent, the kidney 12 per cent, and the spleen, 1.4 per cent of the injected dose. 5. Approximately 20 per cent of the injected peroxidase was excreted in the urine during the first 6 hours, and the concentration of peroxidase in blood serum and urine fell exponentially during this time. After 6 hours, only low concentrations were excreted in the urine but low enzyme activity was still detectable after 30 hours. Approximately 6 per cent of the injected dose was excreted in the feces from 6 to 20 hours after administration. 6. After feeding through a stomach tube, low concentrations of peroxidase were found in blood serum and urine. Considerable variations in the extent of absorption from the gastrointestinal tract were observed in individual rats.     

COLORIMETRIC INVESTIGATION OF THE UPTAKE OF AN INTRAVENOUSLY INJECTED PROTEIN (HORSERADISH PEROXIDASE) BY RAT KIDNEY AND EFFECTS OF COMPETITION BY EGG WHITE

After intravenous injection of horseradish peroxidase into rats, the foreign protein appeared in the kidney first in the small phagosomes and its concentration there decreased quickly; it then was concentrated and "stored" for several days in the large phagosomes. After injection of 10 mg of peroxidase per 100 gm of body weight, the concentration of peroxidase in blood and urine decreased exponentially during the first 6 hours; small amounts of peroxidase were excreted in the urine for several days. When 0.05 to 1.0 mg of peroxidase per 100 gm were administered, most of the peroxidase was taken up by the liver and little by the kidney, and a portion was excreted in the urine even at the lowest dose. At doses above 1.5 mg per 100 gm, the liver cells were saturated, and large reabsorption droplets appeared in the tubule cells of the kidney. With further dosage increase, the concentration of peroxidase in the phagosomes of the kidney increased rapidly until saturation was reached at doses of 13 mg per 100 gm. After intraperitoneal injection of egg white 18 hours prior to the administration of peroxidase, the concentration of peroxidase in all kidney fractions was only 10 to 25 per cent of the values for the untreated animals, the disappearance of peroxidase from the blood was delayed, and 81 percent more peroxidase was excreted in the urine. The treatment with egg white had no effect on the uptake of peroxidase by the liver. The ability of kidney tissue to degrade and adsorb peroxidase in vitro was tested.     

Efficacy of meropenem in the treatment of severe complicated urinary tract infections]

The clinical and bacteriological efficacies of meropenem in the treatment of 12 patients with urinary tract infection were studied. In 8 patients the drug was administered intravenously in a dose of 1 g every 8 hours and in 4 patients with the creatinine clearance below 50 ml/min it was administered in a dose of 1 g every 12 hours (the treatment course of 7 to 10 days). Meropenem was used in the monotherapy. Severe complicated urinary tract infections were mainly observed in the patients with long-term urolithiasis, subjected to repeated surgical interventions and isolating as a rule polyresistant strains of Pseudomonas aeruginosa and E.agglomerans as the pyelonephritis pathogens at a titre of 5 x 10(5)-5 x 10(8) microbial cells per 1 ml of the urine susceptible to meropenem in 80 to 96 per cent of the cases. The clinical efficacy of the drug was stated in all the patients while the bacteriological efficacy amounted to 88.9 per cent.     

A SURVEY OF THE RESULTS OF TREATMENT OF GASTRIC CANCER IN SAN FRANCISCO

A survey was made of all patients treated for gastric cancer on the clinic services of the Stanford University Hospital during the 30-year period 1919 to 1948. During the last decade of the survey there were impressive gains in the surgical treatment of this disease. It was possible from 1944 to 1948 to do a gastric resection on half the patients seen with cancer of the stomach. Also, there was a pronounced decrease in resection mortality so that from 1939 to 1948 the mortality rate for subtotal gastrectomy for cancer was 3 per cent.The over-all five-year survival rate was discouragingly low—4.6 per cent. On the other hand, 23 per cent of those surviving gastric resection lived for five years.A survey of the management of carcinoma of the stomach from 1939 to 1948 was made in 11 general hospitals in San Francisco. A wide range of resectability and resection mortality rates was observed. The cases from these hospitals were combined with those from Stanford for the same period to form a composite group of 1,128 patients. Analysis of this group of cases from 12 representative hospitals in San Francisco showed encouraging trends toward higher resectability rates with a lower resection mortality.     

6-Keto-prostaglandin F1 alpha is not added to urine during passage through the rat urinary bladder in vivo

Studies were conducted to determine whether prostaglandins are added to the urine during its passage through the rat urinary bladder in vivo. Control rats and rats with chronic streptozotocin-induced diabetes were anesthetized with Inactin, 100 mg/kg i.p., and urine was collected simultaneously from both kidneys. Urine from the left kidney was collected directly from the renal pelvis via a ureteral cannula, while urine from the right kidney was collected via a cannula in the urinary bladder. Prostaglandins in the urine were measured by radioimmunoassay. No difference in urinary concentration or rate of excretion of 6-keto-PGF1 alpha or PGE2 was seen between ureteral urine and bladder urine from either normal or diabetic rats. The results of this study indicate that in vivo there is no intralumenal addition of either 6-keto-PGF1 alpha or PGE2 to the urine by the ureteral bladder of rats.     

Cardiac screening procedures

Seven per cent of a sample group of civil service employees were found to have diagnosable heart disease. The diagnosis was made on the basis of a complete physical examination and history taken by a specialist in internal medicine and/or heart disease. In addition a questionnaire form related to symptoms of heart disease was filled out by the patient, and various laboratory tests were made. As a technique in cardiac case-finding, the electrocardiogram was the best single device. Of all the cases of heart disease in this survey 65 per cent were detected from tracings obtained by using all 12 leads, and 57 per cent if only the three standard limb leads were taken; but of the presumably normal persons, 13 per cent would be erroneously suspected of possible heart disease by this technique. Heart disease was detected in 50 per cent of the diagnosed cases on the basis of answers to three of the questions in the questionnaire. Eighteen per cent of normal persons would also have been suspected of having heart disease by this case-finding device. Although the survey reported did not develop a simple cardiac case-finding technique, the data presented indicated that a questionnaire history-form, if judiciously used and evaluated, may be of value to physicians who examine large numbers of patients who are unwilling to submit to a complete cardiac evaluation.     

Antibacterial prophylaxis of bacteriuria at transrectal multifocal prostatic biopsy]

It was shown that prophylactic use of ciprofloxacin (500 mg per os 30 min prior to and 5 days after transrectal multifocal prostatic biopsy) along with topical treatment with 40 ml 1% povidone-iodine and evacuant enema provided negative bacteriological urine analysis in 24 hours for 94.4 per cent of cases. Positive effect was registered for all patients as no urinary tract infections were demonstrated. Transitory fever over 37.5 degrees C was not registered at 67 (97.2 per cent) patients, for the rest cases no changes of the treatment regime were necessary. The results of the trial proves high bacteriological and clinical efficacy of the therapy regimes and allow to recommend its implementation at transrectal biopsy.     

Antibiotic resistance dynamics of plasma-coagulating staphylococci isolated from patients in 1970-1975]

Resistance of 2345 strains of plasmocoagulating staphylococci isolated from purulent inflammatory foci of surgical patients was studied with respect to the widely used antibiotics by the method of standard paper discs in 1970--1975. It was noted that the cultures resistant to erythromycin and monomycin were more frequent, i.e. from 24.2 +/- 2.5 to 51.4 +/- 3.4 per cent and from 1.0 +/- 0.6 to 28.0 +/- 2.1 per cent respectively, p less than 0.001 in both cases, while the percentage of the strains resistant to benzylpenicillin and tetracycline steadily increased, i.e. from 69.9 +/- 2.4 to 47.0 +/- 2.3 per cent and from 72.8 +/- 2.4 to 28.4 +/- 2.1 per cent respectively, p less than 0.001 in both cases. The number of the resistant cultures to streptomycin and levomycetin slightly changed and was relatively high (about 50 per cent and more). Direct correlation (mean and pronounced) between the amount of levomycetin, tetracycline, erythromycin or monomycin used per citizen of Minsk and the frequency of the strains isolated from the patients to these drugs was noted.     

Characteristics of the antibiotic sensitivity of nitrofuran--resistant Gram-negative bacteria of clinical origin]

494 persons with various purulent inflammatory diseases were examined. Gram-negative nitrofuran resistant organisms isolated from the clinical material were found in 22.9 per cent of the cases. Representatives of Enterobacteriaceae, Klebsiella-Enterobacter-Serratia (26.8 per cent), Proteus-Providencia (16.8 per cent) and E. coli-Arizona-Citrobacter (16.1 per cent) predominated. Pseudomonas (32.1 per cent) and Acinetobacter (5.3 per cent) predominated among the aerobic gram-negative bacteria. Sensitivity of 131 strains to 10 chemotherapeutic drugs was determined. 60.4 per cent of the aerobic and 44.2 per cent of the Coli bacteria had multiple drug resistance (to 5 and more drugs). The organisms isolated from the urine and wound excretion were most resistant. Representatives of Pseudomonas and Klebsiella-Enterobacter predominated among the polyresistant cultures. High sensitivity of the isolates to gentamicin, carbenicillin and nalidixic acid was noted.     

Retroperitoneal tumors in children; roentgen diagnosis

A study was undertaken to determine whether there are any features of retroperitoneal tumors in children that might be demonstrated on roentgenograms to aid in identifying them preoperatively. Study was limited to Wilms' tumor of the kidney and neuroblastoma. Calcification was found in 57 per cent of the neuroblastomas and in only 12 per cent of Wilms' tumors. Calcifications in neuroblastomas differed from those in Wilms' tumors. Calcification in neuroblastoma was more frequent in older children than in the younger ones. The kidney was frequently displaced by both types of tumor. However, the neuroblastoma always displaced the kidney downward, or downward and slightly outward. In most instances, the Wilms' tumor also displaced the kidney downward and outward, but in some instances upward and medially. This, of course, depended upon the site of origin of the tumor. There was a distortion of the intrarenal structures in 75 per cent of the cases of neuroblastoma and in 71 per cent of the cases of Wilms' tumor.     

6-keto-prostaglandin F1α is not added to urine during passage through the rat urinary bladder

Studies were conducted to determine whether prostaglandins are added to the urine during its passage through the rat urinary blader . Control rats and rats with chronic streptozotocin-induced diabetes were anesthetized with Inactin, 100 mg/kg i.p., and urine was collected simultaneously from both kidneys. Urine from the left kidney was collected directly from the renal pelvis via a ureteral cannula, while urine from the right kidney was collected via a cannula in the urinary bladder. Prostaglandins in the urine were measured by radioimmunoassay. No difference in urinary concentration or rate of excretion of 6-keto-PGF_1α or PGE₂ was seen between ureteral urine and bladder urine from either normal or diabetic rats. The results of this study indicate that there is no intralumenal addition of either 6-keto-PGF_1α or PGE₂ to the urine by the ureteral bladder of rats.     

The role of external irradiation in the treatment of transitional cell carcinoma of the bladder-a review of fifty cases

Of 100 patients with carcinoma of the bladder seen in the Section of Therapeutic Radiology, University of California, San Francisco, between 1957 and 1962, 59 were accepted for radiation treatment. Fifty had transitional cell carcinoma and were treated with supervolt therapy (1 mev or cobalt-60).TWO TYPES OF TUMORS WERE AGAIN FOUND SUITABLE FOR EXTERNAL IRRADIATION: Papillary carcinomas Grades II and III, as long as they have not, or at least have not massively, invaded muscle; and undifferentiated carcinomas, Grade IV, regardless of degree of extension through the pelvis. The former type, if single, is treated by irradiation for the first recurrence after one attempt with radical transurethral resection. In the presence of multiple lesions at the first examination, radiation therapy is given immediately. The latter type is treated by radiation therapy without any attempt at surgical removal.Of 37 patients, Stages A to C, treated more than three years ago, 14 (38 per cent) lived more than three years and eight (22 per cent) had no cystoscopic or clinical signs of active disease and had normal bladder function. Of 23 patients treated more than five years ago, eight were alive after five years (35 per cent) and four (17 per cent) remained controlled by radiation therapy alone, with normal bladder function.No major complications were observed. In particular, no fibrosis of the bladder occurred. Doses ranged from 5,000 r in five and a half weeks to 6,000 r in seven weeks.A close cooperation between urologic surgeons and radiotherapists during recent years permits long-range treatment planning from the time of diagnosis, which is essential in the effective therapy of carcinoma of the bladder.     

The Role of External Irradiation in the Treatment of Transitional Cell Carcinoma of the Bladder—A Review of Fifty Cases

Of 100 patients with carcinoma of the bladder seen in the Section of Therapeutic Radiology, University of California, San Francisco, between 1957 and 1962, 59 were accepted for radiation treatment. Fifty had transitional cell carcinoma and were treated with supervolt therapy (1 mev or cobalt-60).Two types of tumors were again found suitable for external irradiation: Papillary carcinomas Grades II and III, as long as they have not, or at least have not massively, invaded muscle; and undifferentiated carcinomas, Grade IV, regardless of degree of extension through the pelvis. The former type, if single, is treated by irradiation for the first recurrence after one attempt with radical transurethral resection. In the presence of multiple lesions at the first examination, radiation therapy is given immediately. The latter type is treated by radiation therapy without any attempt at surgical removal.Of 37 patients, Stages A to C, treated more than three years ago, 14 (38 per cent) lived more than three years and eight (22 per cent) had no cystoscopic or clinical signs of active disease and had normal bladder function. Of 23 patients treated more than five years ago, eight were alive after five years (35 per cent) and four (17 per cent) remained controlled by radiation therapy alone, with normal bladder function.No major complications were observed. In particular, no fibrosis of the bladder occurred. Doses ranged from 5,000 r in five and a half weeks to 6,000 r in seven weeks.A close cooperation between urologic surgeons and radiotherapists during recent years permits long-range treatment planning from the time of diagnosis, which is essential in the effective therapy of carcinoma of the bladder.     

Various methods of using lincomycin for treating acute and chronic hematogenic osteomyelitis in children]

Comparative data on the treatment of 209 children with acute and chronic hematogenic osteomyelitis are presented; 128 patients hospitalized before 1974 were treated with antibiotics, mainly penicillin and streptomycin without sensitivity testing. From 1974 81 children were treated with lincomycin; 80 per cent of the isolates were sensitive to this antibiotic. In lincomycin therapy the method of electrophoresis on the disease focus, intrabone administration of the drug and administration of the drug into the bone cavity together with the blood clot during surgical interventions in cases with chronic hematogenic osteomyelitis were used. A marked decrease in the rate of the chronic forms of the disease was registered (from 77.2 to 8.8 per cent).     

SUMMER DIARRHEA IN THE SAN JOAQUIN VALLEY

In laboratory, epidemiologic and clinical studies of 85 patients with diarrhea admitted to the General Hospital of Fresno County and the San Joaquin County Hospital during part of the summer of 1949 the following features were noted:1. Cultures were positive for Shigella in about 45 per cent of the cases in the San Joaquin County Hospital and in about 15 per cent of those in the Fresno County Hospital.2. In 80 per cent of cases in which there was gross blood in the stools before the third day of hospitalization, Shigella grew on cultures. No gross blood was noted in 27 per cent of cases in which Shigella was demonstrated.3. The cases in which Shigella was demonstrated did not differ greatly from the others with regard to fever, leukocytosis, or response to therapy.4. Salmonella organisms were not grown on cultures in any case.5. Most of the patients were infants younger than one year of age.6. Poor socio-economic and hygienic conditions characterized the households from which these county hospital patients came.7. Household follow-up studies indicated that there had been one or more instances of diarrhea among household associates of approximately half the hospitalized patients at some time during the month prior to admission.     

Stapes mobilization for otosclerotic deafness; the monitored peribasal technique

Direct surgical operation on the stapes footplate region in the treatment of hearing loss due to otosclerosis has been revived. In the last 100 cases of a total series of 600, the peribasal stapes mobilization (stapedolysis) technique was completely successful in 62 per cent, partially successful in 29 per cent and a failure in 9 per cent. Stapes mobilization requires precise microsurgical technique monitored by audiometry during the surgical procedure. It appears that mobilization is the preferred approach to the surgical treatment of otosclerosis, but fenestration of the vestibular labyrinth is an important secondary procedure in some cases. Successful restoration of hearing requires adequate mobility of the footplate coupled with physiological continuity of the drum ossicular chain mechanism.     

CIRRHOSIS OF THE LIVER—Clinical Course in 2,377 Patients at San Francisco General Hospital

The records of 2,377 patients with Laennec''s cirrhosis were reviewed for the period 1947-1957. The chief presenting symptom was ascites in 46 per cent, bleeding in 23 per cent, coma in 18 per cent, jaundice in 9 per cent, and both jaundice and ascites in 4 per cent. Nearly half of the patients died during the period under study—one-third from hepatic failure, one-third from gastrointestinal bleeding, and one-third from other causes, most of which were related to alcoholism.Massive gastrointestinal bleeding occurred in 21 per cent of the patients at some time in their clinical course, and in the 10 per cent of these in whom ulcer was demonstrated, one-fifth died as a result of the hemorrhage. Of those presumed to be bleeding from esophageal varices, 64 per cent died at the first hemorrhage and 10 per cent at subsequent hemorrhages; 85 per cent of all those who bled from varices were dead at the end of one year, and 91 per cent were dead at the end of three years.The survival curve of a group of patients who bled once and were good operative risks but had received no operative treatment was compared to the survival curve for entire group who survived the first hemorrhage. The three-year survival in the good risk group was 47 per cent; for the group as a whole it was 30 per cent. The difference in mortality rate was primarily due to an increased number of deaths from hepatic failure in the combined group, whereas 60 per cent of the good risk group died of recurrent gastrointestinal hemorrhage.As 86 per cent of those who were to die of gastrointestinal bleeding did so at the first hemorrhage, it was concluded that any decided improvement in the salvage rate achievable by operation must come from some means of diagnostic forecast of the likelihood of bleeding, with resort to prophylactic operation in such cases.     

RETROPERITONEAL TUMORS IN CHILDREN—Roentgen Diagnosis

A study was undertaken to determine whether there are any features of retroperitoneal tumors in children that might be demonstrated on roentgenograms to aid in identifying them preoperatively. Study was limited to Wilms'' tumor of the kidney and neuroblastoma.Calcification was found in 57 per cent of the neuroblastomas and in only 12 per cent of Wilms'' tumors. Calcifications in neuroblastomas differed from those in Wilms'' tumors. Calcification in neuroblastoma was more frequent in older children than in the younger ones.The kidney was frequently displaced by both types of tumor. However, the neuroblastoma always displaced the kidney downward, or downward and slightly outward.In most instances, the Wilms'' tumor also displaced the kidney downward and outward, but in some instances upward and medially. This, of course, depended upon the site of origin of the tumor.There was a distortion of the intrarenal structures in 75 per cent of the cases of neuroblastoma and in 71 per cent of the cases of Wilms'' tumor.     

Prolonged antibacterial action of polymer coated suture materials]

The antimicrobial activity of capromed, a surgical polymer-coated sutural material containing dioxidine, quinoxidine or gentamicin was studied in vitro and in vivo. Capromed was shown to be active against the hospital strains of S. aureus, E. coli, Proteus spp., Klebsiella spp. and P. aeruginosa. The antimicrobial activity of ADH and AG threads was preserved for 3 to 4 days after implantation. The DH-2 and G-2 threads preserved their activity for 6 to 7 days. It was concluded that the duration of thread antimicrobial activity depended on the properties of the polymer coating the thread. Capromed was applied to 280 operation wounds in 275 patients. There was no wound suppuration in the group of patients after pure operations (n = 62). In the group of patients after conditionally pure operations (n = 130) suppuration was observed in 2 patients (1.3 per cent). In patients with contaminated wounds (n = 88) suppuration in 5 of them (5.7 per cent) was recorded. The total number of the purulent complications after using capromed in surgical operations amounted to 2.5 per cent. In the control group purulent complications were stated in 8.2 per cent of the cases.