Hemidiaphysectomy to control chronic osteomyelitis.

A retained Küntscher nail in the left femur of a 34-year-old man was complicating the course of chronic, unremitting osteomyelitis. The nail was bent and could not be removed by routine methods. A hemidiaphysectomy of the femur allowed easy removal of the nail and radical curettage of the chronically infected intramedullary granulation tissue. The portion of the diaphysis removed at osteotomy was replaced without internal fixation. Postoperative external fixation was considered unnecessary because of the bulk of the femoral diaphysis secondary to the chronic sclerosis of osteomyelitis. Roentgenograms taken seven months after operation showed healing of the femur and no evidence of sequestration of the bone fragment. At 23 months' follow-up, the patient has no pain, no drainage, and no limitation of activities. Long-term follow-up is planned.


The halo vest: an evaluation of motion and forces across the neck.

Flexion-extension motion and compression-distraction forces across the cervical spine were evaluated in 6 patients in halo vests and in 1 patient in a halo cast. Motion in the vest was evaluated in supine and upright positions and averaged 31% of normal motion at the levels tested. Compression-distraction forces showed great individual variation and frequent compression. The average distraction force varied in different positions by nearly 20 pounds in the vest and over 30 pounds in the cast. Methods of improving cervical spine stability in halo orthoses are discussed.


Odontoid fracture complicating ankylosing hyperostosis of the spine.

Ankylosing hyperostosis of the spine (Forestier's disease) is a distinct clinical entity which must be differentiated from ankylosing spondylitis (Marie-Strumpell's disease) and hypertrophic spondylosis. In the case presented, the distinction was made by clinical, roentgenographic, and postmortem pathologic findings. This case had the unique and previously unreported feature of complication by a fracture of the odontoid. The patient's particular circumstances led to the decision to treat the fracture by posterior arthrodesis, though this is not necessarily recommended for all spinal fractures occuring in this disease.


Healed untreated anterior cervical spine dislocation. A case report.

Fracture and fracture-dislocations of the cervical spine are serious injuries that rarely go undiagnosed. A case report of a bilateral facet dislocation with anterior displacement is presented that went undiagnosed and untreated, and healed without neurologic sequelae. No previous similar reports are known.


Vertebra plana and eosinophilic granuloma of the cervical spine in children.

Eosinophilic granuloma of the cervical spine in children is a rare lesion which presents both diagnostic and therapeutic problems. The report describes three additional cases. Open biopsy is recommended instead of attempted needle aspiration in children. Laminectomy is likely to result in spinal deformity, and is not advised unless a second stage fusion is planned. Radiotherapy is effective in controlling the lesion. The ultimate outcome is usually good.


Energy expenditure during walking in patients with scoliosis. The effect of surgical correction.

Oxygen uptake during treadmill walking was measured in 43 female patients with scoliosis, aged 11 to 32 years. The majority of the patients had idiopathic thoracic curves, ranging from 25 to 160 degrees. The study included variables such as trunk imbalance, heart rate, step frequency, and respiratory data. Compared to age- and sex-matched controls, the scoliotic patients showed a significantly higher oxygen uptake/kg body weight and higher heart and respiratory rates. There was a negative correlation between the oxygen uptake/kg and the vital capacity. Trunk imbalance or palvic obliquity, measured from standing radiograms, did not correlate with energy expenditure, nor did the degree of the curvature. Twenty-eight of the patients underwent surgical correction and fusion of the spine with the Harrington technique. Two years after surgery, these patients showed a mean decrease in oxygen uptake/kg, significant at low walking speeds but not at high velocities. A decrease in oxygen uptake/kg was related to an increase in vital capacity. A correlation between a lowered oxygen uptake/kg and a gain in weight was observed.


Combined Dwyer and Harrington instrumentation and fusion in the treatment of selected patients with painful adult idiopathic scoliosis.

Five patients with painful adult idiopathic scoliosis and double structural curve patterns were treated by combined Dwyer and Harrington instrumentation and fusion. The goal of treatment, achieved in all 5 cases, was elimination of pain, stabilization of the spine, and restoration of torso balance. The thoracic curve was corrected an average of 24%, and the lumbar curve, 46%. The loss of correction was only 5 degrees at followup. There were no pseudoarthroses and no complications.


X-ray fluorescence analysis of muscles in scoliosis.

X-ray fluorescence spectrometry was used to determine calcium, copper, and zinc concentrations in paraspinal and gluteal muscles obtained during spinal surgery from patients with scoliosis. Samples of 1-3 mg were sufficient to demonstrate that calcium was higher in idiopathic than in other forms of scoliosis or in normal control muscles. It is suggested that a calcium-related neuromuscular defect could be an important factor in the genesis of idiopathic scoliosis.


Scoliosis in arthrogryposis multiplex congenita.

Fourteen of 50 patients studied with arthrogryposis also had scoliosis, an incidence of 28%. Eight patients developed a severe curve associated with functional disability and poor health. Three patients were unable to walk independently, and 2 died in childhood. Seven patients had congenital scoliosis; 4 had long "C" neuromuscular curves, and 3 had scoliosis associated with pelvic obliquity caused by unilateral hip dislocation or contracture. The natural history of congenital and neuromuscular curves was progression to an extreme and rigid scoliosis. The authors urge others to look for scoliosis in their patients with arthrogryposis and to anticipate its progression.


Occurrence of cold agglutinins in congenital scoliosis.

Eighty-seven patients undergoing spinal surgery for a variety of conditions were surveyed for the presence of cold agglutinins. Patients with congenital scoliosis or metastatic lesions of the spine were found to have a frequency of cold agglutinins that was 250 to 1000 times greater than that in the general population. This greater frequency was independent of ABO blood group, sex, or age. Pre-admission screening of congenital scoliosis patients is urged.


Fracture-dislocation of the dorsal-lumbar spine. Acute operative stabilization by Harrington instrumentation.

Twenty-four patients (of whom 17 were paraplegic) with acute fracture dislocations of the dorsal-lumbar spine underwent stabilization with Harrington instrumentation and were followed for an average of 16 months. Immediate postoperative ambulation was achieved in 16 patients. There was little difference between compression and distraction with respect to reduction and maintenance of correction. Compression rods resulted in solid union in 15 of 16 patients, while distraction rods were successful in 6 of 8 instances. In contrast to reported series in which operative intervention has been avoided, at least a 50% reduction in total hospitalization stay and costs has been achieved. Functional results were comparable to or exceeded the results of other series, and complication rates were similar with the exception of postoperative pain in 2 patients.


Hypotensive anesthesia for spinal fusion with sodium nitroprusside.

Deliberate induced hypotension for spinal fusion has been used in the past, but a search for better techniques continues. The authors used sodium nitroprusside by intravenous infusion to reduce the mean arterial pressure from 72 to 42 torr in 10 cases. This degree of hypotension provided an optimal surgical field and maintained adequate tissue perfusion as demonstrated by arterial oxygen tensions greater than 600 torr, no development of base deficit, and continuous urinary excretion. There were no complications related to hypotension. None of the patients received blood transfusions, whereas in the past, 2000 ml blood was usually required for this procedure.


Microlumbar discectomy: a conservative surgical approach to the virgin herniated lumbar disc.

Microlumbar discectomy is an extremely conservative microsurgical procedure for the treatment of the virgin herniated lumbar disc. The technique has been utilized in 530 patients over a 5 1/2-year period, and features no laminectomy or curettement of the disc space. Preservation of extradural fat and blunt perforation of the annulus fibrosus, rather than scalpel incision, appear to minimize reherniations and adhesion reactions. Operating time has averaged 37 minutes and postoperative stay 3.1 days during the past 36 months (46% of these were compensation cases). In this series, a satisfactory result is defined as those patients who remain economically productive and physically comfortable at the time of followup. Satisfactory results were achieved in 91% of patients after one procedure.


Microlumbar discectomy: followup of 147 patients.

Microlumbar discectomy is a new surgical technique for the treatment of herniated lumbar disc. The operating microscope and special instruments enable the surgeon to remove the herniated portion of the disc without laminectomy through a 1-inch skin incision. A transfusion was never necessary. In a followup study of 147 patients operated on over a 2 1/4-year period, the surgical cure rate was 96%, and the postoperative hospital stay was reduced to less than 3 days. One year after surgery, all noncompensation cases were working as were 80% of the compensation cases. Microlumbar discectomy is safe, effective, and economic for the patient.


A lumbar disc surgery predictive score card.

A lumbar disc surgery predictive score card or questionnaire has been developed to assess potential candidates for excision of a herniated lumbar disc who have not previously undergone lumbar spine surgery. It is not designed to encompass patients who are being considered for other types of lumbar spine surgery, such as decompressive laminectomy or fusion. In an effort to make the "score card" usable by almost all physicians who are involved in lumbar disc surgery, only studies which have broad acceptance and are generally employed are included. Studies which have less widespread use such as electromyogram, discogram, venogram, special psychologic studies (MMPI, pain drawings) have been purposely excluded.


Isolation and partial characterization of drug-resistant Aedes albopictus cells.

We report here the first isolation and characterization of drug-resistant cell lines from cultured mosquito cells. A BrdU-resistant cell line was isolated from mutagen-treated Aedes albopictus cells, and then used as the parent cell from which sublines resistant to ouabain or alpha-amanitin were derived. The BrdU-resistant cell line and all sublines derived from it were shown to be deficient in thymidine kinase. Evidence that favored attributing the drug-resistant phenotypes to mutant genotypes included (a) the drug-resistant phenotypes were stable for up to 16 months of culture under nonselective conditions, and (b) the frequency of the ouabain-resistant cells in a sensitive population was increased 500-fold (to between 10(-3) and 10(-4) by exposure to the chemical mutagen EMS. Evidence illustrating the usefulness of mutant mosquito cells for the study of the genetics of somatic cells is discussed.


Canavanine-resistant variants of human lymphoblasts.

Variants resistant to canavanine, an arginine analogue, have been isolated from two long-term human lymphoblastoid cell lines. They are 20-fold more resistant to canavanine than the parental lines and this phenotype is stable in the absence of canavanine for more than 100 generations. The specific activity of argininosuccinate synthetase, the first of two enzymes necessary for the conversion of citrulline to arginine, is elevated in variants from both cell lines. Furthermore, this enzyme activity is refractory to the repression caused by arginine in normal lymphoblasts. The specific activity of argininosuccinate lyase, the second enzyme in the pathway from citrulline to arginine, is not appreciably changed. Arginine uptake appears normal in the variants since they grow as well as the parental lines in media containing a wide range of arginine concentrations. Arginyl-tRNA synthetase activity is also unchanged. Thus the canavanine-resistant variants have altered control of at least one urea cycle enzyme and appear to be regulatory mutants of human cells.


Hypokalemia in resuscitation from multiple trauma.

In a series of 75 patients review for injuries of the gastrointestinal tract, a consistent degree of hypokalemia and hypofibrinogenemia was noted. The degree of hypokalemia was associated with the severity of trauma and subsequent mortality. The degree of hypofibrinogenemia was associated more with mortality than with severity of trauma.


Intracellular sodium and potassium changes in vascular smooth muslce during hemorrhagic shock.

The vascular smooth muscle cell sodium and potassium changes occurring in hemorrhagic shock were studied in rats subjected to hemorrhagic shock at 30 millimeters of mercury for a two hour period. A new ion exchange method, based upon the substitution of lithium for extracellular sodium was used to measure the intracellular sodium and potassium. A significant rise in cell sodium and fall in cell potassium levels occurred. These changes suggest that vascular smooth muscle cell membrane function is impaired in hemorrhagic shock. The normal homeostatic vascular responses to hemorrhage may be significantly affected by these changes.


Long term results following esophageal anastomosis in the neonate.

In this series, a striking decrease in stricture formation following one layer repair of the esophagus in the newborn with esophageal atresia is clearly demonstrated. The long term evaluation of repair of esophageal atresia in the neonate shows a significant association between morbidity and the presence of a stricture. Long term sequelae related to anastomotic leaks were not evident. A close association between a leak and subsequent stricture formation has been suggested, but that relationship is not substantiated by this review. The number of deaths from anastomotic complications is too small to allow a valid comparison, but the mortality for the two anastomotic groups appears comparable. There is also no significant difference in mortality associated with the type of anastomotic complication. On the basis of these results, we suggest that repair of esophageal atresia in the neonate be done using a one layer end-to-end anastomosis through an extrapleural approach with good drainage.


Surgical and pathologic aspects of protein losing gastropathy.

The mechanism of protein loss from the gastric wall was investigated in patients with Ménétrier's disease, gastric polyps, and carcinoma of the stomach. In Ménétrier's disease, plasma protein was lost as glycoprotein in the gastric juice secreted from proliferated foveolar cells. Some of the gastric polyps also showed pathologic findings similar to that of Ménétrier's disease. As a treatment for the diffuse type of Ménétrier's disease, subtotal gastrectomy was recommended by presenting enough recovery from hypoproteinemia. A small amount of protein lost from the remaining portion of the stomach may be almost completely digested and reabsorbed from the intestine. In instances of carcinoma of the stomach, the mechanism of protein loss was speculated to be loss of lymph, blood or other interstitial fluids from opened capillaries of the cancerous surface. Complete removal of the carcinoma, therefore, certainly results in recovery from hypoproteinemia.


High tension electrical injury of the upper extremity.

A group of 18 patients received a high tension electrical injury of the upper extremity. The over-all results were dismal. Opportunity for improvement appears to lie in a regimen of rapid and complete removal of devitalized skin and muscle, avoidance of contamination and desication and the early provision of blood supply and wound coverage with pedicled flap coverage.


Tumor-host interrelationships in carcinoma of the female breast.

Three hundred and two carcinomas of the female breast were studied histologically with special reference to the morphologic aspects of the tumor, its surrounding host tissue and the regional lymph nodes. The nuclear grade of the tumor was positively correlated with the five year survival rate of the patient. Tumor metastases in the regional lymph nodes were observed to be a sign of a poor prognosis. The stromal lymphocyte and mast cell reactions did not correlate with the frequency of nodal metastases or the five year survival rate. Sinus histiocytosis in the lymph nodes was a sign of favorable five year survival because of its presence in cancer-free nodes only. The paracortical activity of the lymph nodes was an important determinant of whether or not tumor metastases appear in the node.


Ascorbic acid requirements in postoperative patients.

The postoperative ascorbic acid requirements of 63 surgical patients were assessed by measurements of buffy layer leukocyte ascorbic acid and the ascorbic acid content of leukocytes. There was a significant reduction in ascorbic acid levels following operation. The postoperative changes were unrelated to the extent of surgical trauma or the volume of blood transfused during operation, but there was a significant correlation between postoperative ascorbic acid measurements and white blood counts. It appears that postoperative leukocytosis and release by the bone marrow of leukocytes with a low ascorbic acid content may partly account for the postoperative changes in buffy layer and leukocyte ascorbic acid measurements. However, surgical operations were followed by an authentic increase in ascorbic acid requirements, and there was a 42 per cent reduction in circulating leukocyte ascorbic acid levels on the third postoperative day. The findings of this study create an argument for the use of ascorbic acid supplements in surgical patients, although it is unlikely that postoperative changes in leukocyte ascorbic acid have pathologic significance in wound repair.


Blood coagulation changes in shock.

A secondary coagulopathy develops in a state of hypovolemic shock; in endotoxin shock, the coagulopathy is primary. As the changes in the hemodynamic and metabolic functions advance in hypovolemic shock, a state of hypercoagulability appears which reinforces the irreversible nature of this condition. In endotoxin shock, a primary mixed coagulopathy develops as an event directly related to the presence of bacterial lipopolysaccharides. The first finding is the activation of the fibrinolytic system through the activation of the serum complement system by means of the participation of the properdin, the so-called alternate pathway to the complement system. At the same time and by an identical mechanism, the activation of Factors VII and XI occurs through the activation of the Fletchner Factor, while the Hageman trait is activated by kinogen. Immediately after and following an antigen-antibody type reaction, endotoxin stimulates activation of platelets in the presence of gamma-2-globulin along the classical pathway of activation of the serum complement.


Venous thromboembolism in the morbidly obese.

Obesity has been considered as a high risk factor in the development of thromboembolism. To test the validity of this hypothesis, the records of 564 morbidly obese patients who underwent gastric bypass for control of their obesity were reviewed. Four patients, 0.7 per cent, had proved fatal pulmonary emboli and three demonstrated detectable, but not fatal, embolization. Fifty-seven patients were studied prospectively with Doppler examinations of the lower extremities, and only one patient had a transient abnormality of venous thrombosis of the calf, which proved to be associated with a nonfatal pulmonary embolus. The morbidly obese patients are not at high risk from thromboembolism, and the prophylactic use of low dose heparin, which may increase known wound morbidity, is discouraged. Doppler ultrasound of the lower extremities, however, has proved usefulness to differentiate deep venous thrombosis of the lower extremity from other causes of pain in the leg of the morbidly obese.


Gas-producing clostridial and nonclostridial infections.

Clostridial infections have always been treated promptly and aggressively on the basis of history and Gram's strain. We believe that nonclostridial infections with similar fulminating, septic courses also should receive as prompt and radical treatment and perhaps more so, as hyperbaric oxygen therapy is of far less benefit in a nonclostridial infection. It is particularly important to treat the high risk, obese, patient with diabetes and an infection of the perineum or groin promptly and aggressively if the current, high morality is to be decreased.


Peritoneolysis in the reconstruction of the Roux-en-Y loop.

A technique to provide an additional 5 centimeters of length, without compromising the blood supply to the anastomosis, was developed. It has been applied to 26 patients, 18 with Roux-en-Y biliary bypass at various levels, five with high Roux-en-Y esophagojejunostomies and three with gastrojejunostomies. In the last three patients with unresectable adenocarcinoma of the distal part of the stomach and invasion of the intestinal mesentery, due to foreshortening of the latter, the proximal loop of the intestine would not reach the desired level of the stomach until this maneuver was performed.


Cimetidine prevents reduction in gastric mucosal blood flow during shock.

This study evaluated Cimetidine's possible role in regulating gastric mucosal blood flow in the anesthetized, stressed miniature swine. Stress consisted of hemorrhagic shock to a mean arterial pressure of 50 mm Hg. Twenty-one animals were divided into three experimental groups: untreated controls, preshock Cimetidine treatment group, and postshock Cimetidine treatment group. Gastric mucosal blood flows were determined (microsphere method) during a stabilization period and after 90 minutes of shock. Central hemodynamic indices were monitored throughout each experiment. In the fundus, mucosal blood flow decreased 59% in the controls, 11% in the preshock, and 28% in the postshock Cimetidine groups. Antral mucosal blood flow decreased 57% in controls, 19% in the preshock, and 33% in the postshock Cimetidine groups. In the corpal mucosa, blood flow decreased 53% in controls, 11% in the preshock group, and 41% in the postshock Cimetidine group. Cimetidine administration, both before and after shock, conferred significant protection on mucosal blood flow changes related to shock. Preshock drug administration had a significantly greater protective effect than postshock treatment on blood flow in the corpal mucosa.


Improved renal function and inhibition of renin and aldosterone secretion following peritoneovenous (LeVeen) shunt.

Twelve patients with cirrhosis, refractory ascites, and varying degrees of renal failure (creatinine clearance, 5 to 44 ml/min) were studied before and up to 2 weeks following peritoneovenous shunt. Creatinine clearance increased 60% or more in seven patients (group I) and 22% or less in five patients (group II). There were no significant differences in maximum urine output or sodium excretion between groups (group I, 4,272 ml/14 hr, 372 mEq/24 hr; group II, 3,722 ml/24 hr, 255 mEq/24 hr). Aldosterone and renin concentrations were higher in group I and showed a greater decrease after shunting. Renin substrate levels also were higher in group I and rose following shunt insertion, while group II remained low. Ascitic fluid was found to contain renin substrate in concentrations of approximately 25% to 50% of plasma concentrations. Patients with the greatest increase in creatinine clearance showed the largest rise in substrate concentration and fall in renin and aldosterone secretion, suggesting a dynamic relationship between these factors. That a diuresis could occur without significant change in these parameters in five of 12 patients suggests independent control mechanisms for renal salt and water excretion and glomerular filtration in the ascitic patient.


The use of medical audits in surgical education.

Quality of medical care audits conducted in the form of a conference attended by surgical residents, faculty, and medical students accomplish the following: (1) provide an excellent educational experience for all participants; (2) are enthusiastically received by all concerned; (3) accomplish considerably more than audits performed in the manner prescribed by the Joint Commission on Accreditation of Hospitals (JCAH); and (4) fulfill JCAH/Professional Standards Review Organization (PSRO) requirements. This modified type of audit can be used successfully as a variant of a teaching seminar in which learning is more predictable because of the active participation of all concerned. Criteria sets which include simple and complex criteria must be developed if audits are to alter patterns of care for complex surgical problems.


Hepatic metabolism in patients with alcoholic cirrhosis.

Fuel homeostasis was studied in 15 patients with hepatic cirrhosis who previously had sustained upper gastrointestinal hemorrhage secondary to portal hypertension. By combining substrate arteriovenous concentration differences with measured hepatic blood flow rates, the exchange rates of metabolites across the liver was calculated. Hepatic extraction of acetoacetate, beta-hydroxybutyrate, lactate, pyruvate, analine, and glycerol was studied. After an overnight fast, splanchnic glucose production in 15 cirrhotic patients was diminished markedly. Despite reduced total glucose production, there was no decrease in hepatic gluconeogenesis; instead, there was increased glucose formation from amino acids, glycerol, lactate, and pyruvate. In patients with hepatic cirrhosis, the liver does not produce as much glucose as does a normal liver; the failing cirrhotic liver is capable of maintaining fuel homeostasis by increased ketone-body production.


The effect of exogenous substrate on hepatic metabolism and membrand transport during endotoxemia.

Endotoxemia in dogs reduced hepatic uptake and biliary excretion of indocyanine green dye. This diminished active membrane transport was associated with reduced hepatocyte membrane potential difference. Studies of arteriovenous concentration differences and flow across the liver demonstrated that endotoxemia increased hepatic glucose and lactate production and decreased oxygen consumption. Correction of this energy deficit occurred following infusion of glucose and insulin, but not after administration of isocaloric quantities of intravenous amino acids. The glucose-insulin infusion during endotoxemia shifted the liver back to an organ of glucose uptake, improved oxygen consumption, and provided the necessary energy for normal dye transport and maintenance of the normal membrane potential difference.


Lung injury following a 50-metre fall into water.

The pulmonary complications of a 50-metre fall to the water (a form of suicide attempt producing 87% mortality) were studied in 15 survivors. Presenting findings included crackles, haemoptysis, and hypotension. The alveolar-arterial oxygen difference was greater than 150 mmHg (20 kPa) in nine subjects on admission. Ventilatory failure developed in 10 of the patients, including all of those with massive haemoptysis. Radiographic findings included pneumothorax and diffuse pulmonary opacities adjacent to the area of impact. Pneumothorax developed within 12 hours of admission in 10 of 15 subjects but was associated with rib fractures in only four subjects. The clinical course of the condition is consistent with the hypothesis that the traumatic pulmonary tears produced interstitial emphysema, with subsequent development of pneumomediastinum, subcutaneous emphysema, and pneumothorax. Pneumothorax is a common complication of severe lung contusion even in the absence of penetrating pleural injury.


An outbreak of pleural mesothelioma and chronic fibrosing pleurisy in the village of Karain/Urgüp in Anatolia.

The 575 inhabitants of the remote Anatolian village of Karain suffered 11 deaths from pleural mesothelioma in 1975/76 and there were five cases of fibrosing pleurisy. In the previous five years there had been 25 cases of mesothelioma. Calcified pleural plaques were common on survey radiography. Asbestos does not occur in the local soil or rock, nor is it handled in the village, but a few fibres were found in the water. Fibres were also found in the pleural tissue of two of five cases examined. Inhabitants of the neighbouring villages are free of mesothelioma.


Peripheral pooling of bronchographic contrast material: evidence of its relationship to smoking and emphysema.

Sixty-six subjects, mainly derived from various occupational groups and one-third of whom admitted to dyspnoea on exertion, have been grouped according to the appearance of their peripheral airways at bronchography with oily propyliodone. Eleven subjects showed marked peripheral pooling of radiographic contrast material, 22 showed mild or moderate pooling, and in 33 peripheral pools were absent. Pooling was not seen in non-smoking subjects. In the group of subjects without pooling, pulmonary function in non-smokers and subjects with a history of smoking was similar. Subjects with marked pooling had a significantly lower pulmonary diffusing capacity (transfer factor) and evidence of loss of pulmonary elastic recoil when compared with subjects with absent peripheral pooling. These results indicate that bronchographic peripheral pooling is associated with the physiological changes of panacinar pulmonary emphysema and suggest that a causal relationship may exist between the organic bronchiolar lesion of pooling and the peripheral parenchymal lesion of panacinar emphysema.


Carboxyhaemoglobin levels and inhaling habits in cigarette smokers.

In 520 men who currently smoked only cigarettes, carboxyhaemoglobin (COHb) levels were measured as a method of estimating the extent to which cigarette smoke was inhaled and the results were compared with the smokers' own assessment of their inhaling habits. The mean COHb level after standardising for the number of cigarettes smoked before the blood test on the day of the test was 4.0% in self-described non-inhalers. This was much higher than the mean level of 0.7% in 1891 similar non-smokers, but not very different from the standardised mean levels of 5.2%, 5.3%, and 5.6% in men who said they inhaled slightly, moderately, or deeply, respectively. The increasing trend in the COHb levels of men in the four self-described inhaling categories (nil to deep) was small but statistically highly significant. The data from this study may help to explain some of the anomalous epidemiological results regarding the relationship between self-described inhaling habits and the development of diseases associated with smoking, such as coronary heart disease and lung cancer.


Unilateral pulmonary collapse in asthmatics.

Five asthmatic patients developed collapse of one lung. Three of the patients were children and three of the five had repeated episodes of atelectasis. Episodes of atelectasis were usually associated with localised chest pain, which was not pleuritic in character, and with breathlessness, but without wheezing. The were not related to clinically apparent respiratory infections or to deterioration of the underlying asthma. The cause is obscure, but re-expansion seems to be hastened by oral corticosteroid therapy.


Bronchial responsiveness to inhaled histamine and isoprenaline in patients with airway obstruction.

In order to examine the hypothesis that bronchial reactivity to non-specific constrictor stimuli is influenced by the resting tone of the bronchial smooth muscle, the airway responses to inhaled histamine solution and inhaled isoprenaline were measured in 19 patients with airway obstruction. There was a significant positive correlation between the size of the constrictor response to histamine and the dilator response to isoprenaline (r = +0.83; p less than 0.01) as measured by changes in specific airway conductance. Patients with asthma showed greater bronchial reactivity to both histamine and isoprenaline than those with chronic bronchitis, although some patients had changes intermediate between the two extremes.


Pulmonary and hepatic granulomatous disorders due to the inhalation of cement and mica dusts.

Hepatic and pulmonary granulomas were recognised in two workers exposed respectively to Portland cement and to muscovite dusts. The pulmonary lesions in the patient exposed to cement consisted of histiocytic granulomas and irregular fibrohyaline scars, and in the patient exposed to mica of a diffuse thickening of all interalveolar septa due to new formation of reticulin and collagen fibres and proliferation of fibroblasts and histiocytes. In the liver the following pathological findings were observed: focal or diffuse swelling of sinusoidal lining cells, sarcoid-type granulomas, and, in the case of mica exposure, perisinusoidal and portal tract fibrosis. Abundant inclusions of the inhaled material were identified within the pulmonary and hepatic lesions by histochemical and x-ray diffraction techniques.


Bronchial carcinoma and long-term survival. Retrospective study of 433 patients who underwent resection.

The long-term follow-up of patients with bronchial carcinoma treated by surgery is presented. Of 471 patients who underwent thoracotomy, the tumour could not be resected in 38 (8%). Sixty-three (13.4%) died within the first four weeks; 125 (28.9%) survived more than five years. A high percentage developed either late metastases, late recurrences, or a second primary lung carcinoma. The results of surgical resection for bronchial carcinoma cannot be considered satisfactory, although resection remains the best treatment even in those patients with an apparently unfavourable prognosis. In spite of reservations regarding retrospective studies, conclusions can be drawn regarding diagnosis, therapy, and prognosis. Questions concerning histological type, size, and site of tumour, and tumour stage can be answered only after an adequate postoperative interval. Five years after operation the patient who has apparently been successfully treated may die from a second primary carcinoma.


Pulmonary function studies in healthy Pakistani adults.

Predicted normal spirometric values have been shown to have significant geographical and ethnic variation. These variations are of epidemiological significance in determining the prevalence of disease and of clinical importance in measuring the effects on pulmonary function of various diseases. A total of 599 men were chosen from employees of a package manufacturer, a general hospital in Lahore, and a village in northern Pakistan; 94 students and staff of a women's college in Lahore were also studied. The forced vital capacity (FVC) was recorded from three satisfactory efforts, and the FVC, one second forced expiratory volume (FEV1), and maximal midexpiratory flow (MMF, or FEF25-75%) were calculated from the best FVC effort. The FVC and FEV1 in men were found to be similar to those of a group of emigrant Pakistanis and a north-western Indian population (Delhi) but higher than populations in south and eastern India. Pakistani women had values similar to those of women in northern India. None of the women smoked and, among Pakistani men, the smokers (285) averaged 6.7 pack years. While the FVC and FEV1 values did not differ between smokers and non-smokers, there was a significant difference in MMF (FEF25-75%) in the two groups. This latter finding corroborates studies on North American populations in which smokers generally have had a higher lifelong cigarette consumption. This confirms the MMF (FEF25-75%) to be a more sensitive test of subtle, asymptomatic changes in pulmonary function than the more widely used FVC and FEV1.


Use of BCG as an immunostimulant in the surgical treatment of carcinoma of lung: a five-year follow-up report.

Sixty consecutive surviving patients treated with subdermal BCG (5 X 10(6) organisms) have been followed up for five years after resection of lung carcinoma. A control group of the previous 60 consecutive surviving patients not treated with BCG was similarly studied. We found no statistical evidence that the administration of BCG by the method described influenced the five-year survival rate, nor has any effective action upon involved lymph nodes or small metastases been demonstrated.


Blood flow and pressure measurements of right coronary artery to left ventricle fistula.

A case of a dilated and tortuous right coronary artery to left ventricle fistula was surgically repaired. Preoperative intracoronary pressure measurements showed a sudden depression of diastolic pressure in the terminal part of the right coronary artery. Intraoperative mean right coronary flow was recorded as 315 ml/min. After repair of the fistula, blood flow measurements showed only 35 ml/min, which suggested that a large amount of blood was stolen by the fistula. Although systolic and diastolic murmurs only were heard in our patient, continuous murmurs have been noticed by some observers. From a study of the preoperative physical findings, intraoperative coronary flow measurements, and other operative findings, the genesis of heart murmurs relating to this type of anomaly is discussed.


Tumour production of alkaline phosphatase in a patient with giant-cell carcinoma of bronchus.

A patient with a giant-cell carcinoma of the bronchus was found to have a raised serum alkaline phosphatase without any evidence of bone or liver involvement. After necropsy the excised tumour was found to be producing alkaline phosphatase. The alkaline phosphatase found in the serum during life was heat-labile but that in the tumour was heat-stable. The significance of this is unknown.


A new method of disinfection of the flexible fibrebronchoscope.

A modified method for rapid sterilisation of the fibrebronchoscope using immersion of the instrument in 0.1% benzalkonium chloride for five minutes followed by a further five minutes' immersion in 96% alcohol was tested both by deliberate contamination of rubber catheters and on bronchoscopes after use in patients. The method proved effective in eliminating a wide range of potentially pathogenic organisms.


[Bone content in mechanically deboned pork (author's transl)].

The KOH method is a useful procedure to determine the bone content of pork. Samples containing a known quantity of added bone were tested by the KOH method. There were no significant differences between the results of the KOH method and the added bone content (P greater than 0.05). The correction for desiccation of clean bone is hard to determine because of the variations in loss of weight as a result of evaporation of water on preparing clean bone. The correction for conversion of hard bone residues into dry bone was determined from bones of fattening pigs, sows and male swine. Sex and age of the pigs were found to have little effect on this factor. It is suggested to base testing of mechanically deboned pork on the determination of hard bone residues. A content of 0.4 per cent is suggested as the maximum permissible residue of hard bone in mechanically deboned pork, which is equivalent to a clean bone residue of approximately 1 per cent.


[Cochlosomosis, a problem in raising waxbills kept in aviaries (author's transl)].

A flagellate, (Cochlosoma sp.) in the intestines of waxbills caused disease and death, particularly in the younger birds. Disturbance of the intestines was the characteristic clinical symptom. Treatment with Emtryl (Specia, 40% dimetridazole), 250 mg. per litre of drinking water for three days, was effective.


[Abnormal respiration with dullness over the thorax in cattle. Due to an extensive lesion of the liver (author's transl)].

The case of a four-year-old cow is reported, in which a very extensive necropurulent lesion was localized in the left half of the liver. When the animal was studied clinically and by X-ray, a space-occupying lesion in the thorax was strongly suspected. The importance of including diseases of the liver in the differential diagnosis of thoracic disorders is stressed.


[What is your diagnosis? (author's transl)].

The clinical and radiographic findings, surgical treatment and post-operative course in a dog with a recurrent fistula in the neck are reported. Although a foreign body was discernible on a survey radiograph, pre-operative contrast radiography was carried out using a watersoluble contrast medium, the advantages of which are discussed. The foreign body, a piece of treebark, was removed at surgery. The fistula did not recur during a post-operative period of ten months.


[Treatment of Salmonella carriers in a number of reptile species (author's transl)].

Housing Tejus contaminated with Salmonella on wire-mesh floors did not have any effect on the degree of contamination. The sensitivity of Salmonella organisms isolated from desert monitors to ampicillin was satisfactory, the serotypes being less sensitive to chloramphenicol, furazolidone, neomycin, streptomycin and oxytetracycline. Intramuscular injection of combined chloramphenicol and ampicillin in tejus and lizards was not successful, nor was oral administration of furazolidone in tortoises. On the other hand, oral treatment with combined ampicillin and chloramphenicol was effective in tortoises.


[Detection of C-strain virus in pigs following vaccination against swine fever (author's transl)].

During and shortly after a mass vaccination campaign against swine fever with C-strain virus, samples were received from five vaccinated herds, which were found positive on examination with the cryostat immunofluorescent technique (IFT). In four cases fluorescence was due to C-virus antigen as determined by rabbit inoculation. The samples concerned were derived from 4 to 13 week old pigs which had been vaccinated 6 to 7 days prior to death. The fluorescence was notably bright and was confined to the epithelial lining of the tonsillar crypts. The persistence of C-virus following vaccination was studied in twelve-week-old pigs susceptible to swine fever. The animals were immunosuppressed by a concurrent treatment with corticosteroids. Virus antigen was detected by immuno-fluorescence in the tonsils and kidneys till 15 days following vaccination. The replication of virus in these organs is believed to be responsible for the spreading of C-virus observed in the field. Isolation of vaccine virus in rabbits was also possible till 15 days after vaccination. It is concluded that vaccination with C-virus may interfere with the laboratory diagnosis of swine fever by the fluorescent antibody technique applied to tissue sections for at least two weeks.


[Johne's disease in goats. Report of a case (author's transl)].

A case of Johne's disease in a goat is reported. The final diagnosis was not established until a post-mortem examination had been made. The fact is stressed that the symptoms described do not only suggest parasitic infection but also Johne's disease.


[Listrophorus gibbus, a fur mite in domestic rabbits (author's transl)].

A case of infection with the fur mite of domestic rabbits, Listrophorus gibbus, is reported. Possible methods of treatment of individual rabbits as well as of colonies of rabbits are reviewed. The presence of Listrophorus gibbus in conjunction with Cheyletiella parasitivorax is also discussed.


Effect of spironolactone on urinary excretion of immunoreactive prostaglandin E in essential hypertension and primary aldosteronism.

To determine the effect of aldosterone antagonist on renal prostaglandin E synthesis, the rate of urinary excretion of immunoreactive prostaglandin E was measured radioimmunologically before and during the oral administration of an aldosterone antagonist, spironolactone, in 5 patients with essential hypertension, 3 with primary aldosteronism and 2 with postoperative primary aldosteronism. Spironolactone was administered at an oral dose of 25 mg 4 times daily for about 1 week. In the control state, the rates of urinary prostaglandin E excretion ranged from 151 ng/day to 4,527 ng/day in essential hypertension. The rates were not augmented in primary aldosteronism but decreased after the removal of an aldosterone producing adenoma. No obvious relationship was observed between plasma aldosterone concentration and the rate of urinary prostaglandin E excretion. On the first day of spironolactone administration, the excretion rates of urinary prostaglandin E were markedly increased independently of basal plasma aldosterone level in all cases except one case of essential hypertension. Urinary prostaglandin E excretion was increased with the concominant increase of urinary Na/K ratio in essential hypertension and primary aldosteronism. After the second day, the augmented urinary prostaglandin E excretion was decreased and the changes of urinary prostaglandin E excretion varied from case to case. These results suggest that synthesis of renal prostaglandin E is not mainly regulated by aldosterone in essential hypertension and primary aldosteronism.


Effects of riboflavin-2',3',4',5'-tetrabutyrate and flavin adenine dinucleotide on the platelet aggregation induced by hydrogen peroxide.

Both riboflavin-2',3',4',5'-tetrabutyrate and flavin adenine dinucleotide (FAD), especially the latter, could inhibit H2O2-induced platelet aggregation. FAD enhanced the glutathione reductase activity of platelets. FAD might exert its inhibitory effect on the H2O2-induced platelet aggregation via the glutathione reductase and peroxidase system.


Damage of DNA and its recovery in AH-109A cells treated with carboquone in vivo.

A single injection of carboquone at a dose of 0.1 mg/kg body weight induced damage of DNA of AH-109A cells as revealed by alkaline and neutral sucrose density gradient centrifugation. A significant decrease in the sedimentation velocity of DNA on alkaline sucrose density gradient centrifugation was observed 15 min after the injection, and it returned to that of control after 60 min. Thereafter, the size of DNA decreased progressively. When the cells were lysed with 2% sodium dodesyl sulfate solution and analyzed on neutral sucrose density gradient centrifugation, a similar change in the sedimentation velocity was observed. The results obtained from the studies of intraperitoneal growth of AH-109A cells pretreated with carboquone in vivo and the survival of host animals well corresponded to the extent of the damage of DNA as revealed by alkaline and neutral sucrose density gradient centrifugation.


Morphological characterization of two established cell lines, T24 and MGH-U1, derived from human urinary bladder carcinoma.

Two cell lines, T24 and MGH-U1, established from human urinary bladder carcinoma showed typically epithelial features. Although the proliferating cell population presented a polymorphic appearance including polygonal and elongated cells, ultrastructural studies failed to identify different types of cells in each cell line. Extensive cytoplasmic organelles, microvilli on the cell surface and cell continuity with the junctional complex were common findings in both the cell lines. Virus-like particle was not detected. Chromosome numbers of these cell lines were distributed from hypertriploidy to hypertetraploidy. T24 cell line contained a large number of elongated cells and well developed microvilli as compared with MGH-U1 cell line. The present results suggest that T24 cell line has been transformed to a relatively uniform cell population during the long-term culture as referred to in a previous report.


Structure of a novel sialooligosaccharide from the urine of a patient with mucolipidosis.

A mannose-containing sialooligosaccharide has been isolated from the urine of a patient with a newly recognized mucolipidosis which showed a low liver beta-galactosidase activity and hyperglycopeptiduria. The chemical structure has been determined by glycosidase digestion, Smith degradation and permethylation studies. The following structure has been given for the oligosaccharide: NANAalpha2-6Galbeta1-4GlcNAcbeta1-2Manbeta1-3Manbeta1-4GlcNAc. This compound appears to be derived from an incomplete catabolism of glycoproteins with aspartylglucosylamine type side chains.


Simultaneous determination of plasma 11-deoxycorticosterone, 18-hydroxy-11-deoxycorticosterone, and aldosterone in man.

A method for simultaneous measurement of 11-deoxycorticosterone (DOC), 18-hydroxy-11-deoxycorticosterone (18-OH-DOC) and aldosterone using 1.0-2.0 ml of plasma has been developed. The present method consists of extracting plasma with dichloromethane, separating the DOC, 18-OH-DOC, and aldosterone from other steroids on a Sephadex LH-20 column, and quantitating each steroid by radioimmunoassay. This method was demonstrated to be sensitive, accurate and precise. In 20 normal male subjects, the mean recumbent level of DOC was 9.1 +/- 3.1 ng/100 ml, on random diet, at 0800 h. The corresponding levels of 18-OH-DOC and aldosterone were 8.2 +/- 3.9 ng/100 ml, and 6.7 +/- 2.6 ng/100 ml, respectively. Plasma levels of these three steroids were measured in several types of adrenocortical disorders associated with hypertension and hypokalemia. Patients with Cushing's syndrome due to adrenocortical hyperplasia, and 17alpha-hydroxylase deficiency had elevated DOC and 18-OH-DOC levels, but showed normal or lower aldosterone levels. Hypersecretion of DOC and 18-OH-DOC may cause the symptoms of hypertension and hypokalemia. Patients with primary aldosteronism had elevated levels of DOC and 18-OH-DOC as well as aldosterone. The former two steroids may be hyperproduced as a precursor of aldosterone.


Effects of insulin and oral antidiabetic agents on the plasma triglyceride levels in lipoatrophic diabetes.

Three cases of congenital type of lipoatrophic diabetes were treated with oral antidiabetic agents or insulin for high blood sugar. The plasma triglyceride levels, determined after overnight fasting, were elevated following administration of oral agents or injections of insulin. On these medications, plasma phospholipid and cholesterol also tended to elevate. Paper electrophoresis of plasma revealed an increment of pre-beta-lipoprotein. The levels of plasma triglyceride were reduced when doses of oral antidiabetic agents or insulin were kept constant for several days, and further reduction was observed after withdrawal of these therapeutics. These observations suggest that insulin enhances production and secretion of triglyceride in the liver.


Annulate lamellae in erythroblasts in the mice splenic colonies.

Lethally irradiated mice received a transfusion of normal bone marrow cells from the same strain mice. The transfused colony-forming cells (stem cells) were settled in the spleen of the recipient and proliferated in it into erythroblast colonies. Some of these mice were given chloramphenicol or thiamphenicol for 7 days after marrow cell transfusion. Annulate lamellae were frequently observed exclusively in the erythroblasts of the mice received a injection of thiamphenicol.


Chronic hepatitis B and hepatitis B surface antigen carriers in university students.

A study was made of latent hepatic diseases in university students. 1.4% to 1.6% of the students were positive for hepatitis B surface antigen (HBsAg), and 10.3% for anti-HBs. Of 28 students with HBs-antigenemia, 2 had chronic persistent hepatitis, and 3 minimal hepatitis, 23 being healthy carriers. Hepatitis B e-antigen (HBeAg) was detected in 44% of the students with HBsAg, and anti-HBe in 13%. Anti-HBe was significantly more frequently found in female students with HBsAg than in male students. Though most of the students with HBsAg had high titer of antibody to hepatitis B core antigen (anti-HBc), there were a small number of cases showing low titer. HBsAg and anti-HBs was detected in the same serum specimens of 2 carrier students. Liver damage was also found in 3 students without HBs-antigenemia.


Septicemia caused by Klebsiella oxytoca.

Klebsiella oxytoca was isolated from the blood of a leukemic patient. The typical pathogenicity of Klebsiella oxytoca was observed in the patient. The pathogenicity of several Klebsiella oxytoca strains isolated from blood, urine and sputum of different patients was examined in mice, and it was found that Klebsiella oxytoca isolated from the blood had a higher toxicity than the others.


Studies on brain lesion by administration of monosodium L-glutamate to mice. I. Brain lesions in infant mice caused by administration of monosodium L-glutamate.

Light-microscopic examination was performed on the brain lesions induced by monosodium L-glutamate (MSG) in neonatal and infant mice of ICR strain. Lesions characterized as cytoplasmic balooning, chromatin clumping, pyknosis and karyorrhexis of neurons were recognized in the arcuate nucleus (AN), subfornical organ, preoptic area, area postrema and cerebral cortex. The most vulnerable region was the AN in which the region near the root of  the median eminence was easily damaged. The changes in the AN were severest in 7-day-old mice, but only slight in 20-day-old mice. Thresholds of inducing AN lesions in 10-day-old mice after intraperitoneal injection and force-tube feeding were 0.4 and 0.7-0.8 g/kg body weight, respectively. The threshold of retinal changes was about 2.5-fold that of AN in force-tube feeding. In neonatal mice injected daily with 4 g MSG/kg body weight, the neurons of the AN disappeared almost completely by the 4th day of intraperitoneal administration.


Studies on brain lesions after administration of monosodium L-glutamate to mice. II. Absence of brain damage following administration of monosodium L-glutamate in the diet.

The effects of monosodium L-glutamate (MSG) on the brain of mice were examined by light-microscopy under the intended conditions of use of this material. Basal diet containing 5, 10 or 15% w/w MSG was fed to pregnant, lactating and weanling mice ad libitum, resulting in the ingestion of 14.0, 42.8 or 42.0 g/kg body wt. of MSG a day respectively. Mice at the same developmental stages were given the basal diet and 5% w/w MST in aqueous solution ad libitum, the maximum ingestion of MSG being 16.9, 31.4 and 26.8 g/kg body wt. a day respectively. Light-microscopy examination revealed no pathological changes in the hypothalamic arcuate nuclei (AN) of all treated mice, i.e., pregnant females and their fetuses, lactating females and their sucklings, and weanlin mice. Pregnant females and their fetuses, and weanling mice, injected subcutaneously with either 5.0 or 4.0 g/kg body wt. of MSG, showed "Glu-type lesions" in the hypothalamus.


Effect of hexachlorophene intoxication on learning in rats.

Neurotoxicity in the form of hindlimb paralysis is known to be associated with brain lesions characterized by vacuolation of the white matter following hexachlorophene (HCP) intoxication; these paralytic and histopathological effects are reversible with time after discontinuance of HCP exposure. The rates of acquiring a bar-pressing escape and avoidance behavior were measured in female rats after recovery from paralysis following daily oral dosing with HCP (25 mg/kg). Learning deficits were seen as a delay in reaching a 90% escape performance level, as an increase in the number of sessions required to reach a 50% avoidance criterion, as an increase in training time needed to transfer from escape to avoidance responding, and by an increase in the number of sessions needed to reach a maximum level of avoidance behavior over a 35-session period. Peak responding eventually reached a level comparable to controls only after prolonged periods. Brain lesions seen 2-3 months after HCP intoxiciation correlated with the lag in learning ability but not with the maximum avoidance acquisition before sacrifice. These results indicate a long-lasting behavioral deficiency following neurotoxicity which is slowly reversible.


Toxicologic studies of N-trichloromethylthio-4-cyclohexene-1,2-dicarboximide (captan): its metabolism by rat liver drug-metabolizing enzyme system.

The degree of toxicity caused in rats by captan (N-trichloromethylthio-4-cyclohexene-1,2-dicarboximide) administered intraperitoneally is greater than that induced by orally administered captan. With regard to its effect on the drug-metabolizing enzymes of rat liver, the activity of aniline hydroxylase and the level of cytochrome P-450 were found to decrease in the treated rats 24 h after a single oral dose (650 mg/kg). The loss was even greater in the animals receiving diethyl maleate 1 h prior to captan. Furthermore, usual increase in the activity of drug biotransformation enzymes seen after phenobarbital treatment appears to decrease in rats dosed with this funaicide. In vitro incubations of rat liver microsomes with captan resulted in a profound loss of cytochrome P-450 and the acitivty of benzphetamine N-demethylase as well as aniline hydroxylase. Although the inhibition of drug-metabolizing enzyme activity by captan was observed in microsomal incubations with or without NADPH, a detectable amount of carbonyl sulfide (COS) was found only in the incubations that contained captan plus NADPH. Carbonyl sulfide appears to arise from a captan-derived metabolite, thiophosgene (CSCl2), which decomposes to COS in aqueous solutions and in the presence of NADPH inhibits the activity of drug biotransformation enzymes.


An unbiased comparison of organ weights when an inequality in body weight exists.

It is demonstrated in simulations that, if adjustment for inequality in body weight is to be made by covariance analysis and the comparison is carried out in terms of adjusted means of organ weights thus obtained, the test of significance for an organ weight change can be unbiased, or the chances for detecting spurious effects on organ weights remain at about 5% and the chances for detecting treatment effects are far above the 5% irrespective of differences in body weight. Since the use of relative organ weights frequently violates the unbiasedness, it is concluded that if a treated group differs from the control in body weight the comparison of adjusted means of organ weights by covariance analysis will be far more justifiable than that of relative weights.


Influence of sex hormones on free erythrocyte protoporphyrin response to lead in rats.

Following oral administration of lead a difference in free erythrocyte protoporphyrin (FEP) increase was found between adult male, adult female and suckling rats: young animals are more susceptible than adult female rats which in turn exhibit a greater FEP increase than adult male animals. This observation parallels that made previously in humans. Possible difference in iron metabolism does not appear to explain the sex-linked difference in FEP response to lead. Sex hormones mainly progesterone, seem to play a role but their interaction with lead on the FEP response is restricted to female rats and apparently is not mediated through changes in delta-aminolevulinic acid synthetase activity.


Distribution of endosulfan in plasma and brain after repeated oral administration to rats.

Rats were fed endosulfan (5 or 10 mg/kg) containing alpha- and beta-isomers in the ratio of 2:1, daily for 15 days. The distribution pattern of endosulfan, its isomers and metabolite, endosulfan sulfate, was estimated in the plasma and brain of the rats. On day 16, the alpha-isomer in rats receiving 5 mg/kg was highest in the cerebrum (3.76 microgram/g) followed by the remaining part of the brain (2.66 microgram/g), and the cerebellum (2.04 microgram/g). The concentration of the beta-isomer was 0.06 microgram/g in the cerebrum and 0.02 microgram/g in the cerebellum; no beta-isomer was detected in the remaining part of the brain. The plasma concentration of alpha- and beta-isomers was 2.26 and 0.46 microgram/ml, respectively. No metabolite other than endosulfan sulfate was detected in plasma. No significant changes in brain tissue were observed in any of the groups under treatment. On day 30 (15 days after the last treatment), the concentration in plasma declined more rapidly than that in the brain tissue. At a higher dose (10 mg/kg), the distribution pattern of isomers and its metabolite, endosulfan sulfate, followed almost the same trend except that the concentration was higher than in rats receiving lower doses.


Lower azure B methylene blue ratios in Giemsa type blood and malaria stains.

Starting from ancient reports that rare samples of methylene blue were apparently sufficiently contaminated with azures to give red plasmodial and red purple nuclear chromatin in Chenzinsky type methylene blue eosin stains, it was decided to determine how little azure B would suffice for such staining in methylene blue eosin stains. The traditional 1902 Giemsa had an azure : methylene blue : eosin ratio of about 6 : 3 : 6.3 : 10; Lillie's 1943 formula had a 5 : 7 : 10 ratio. In the current series of tests 5 : 7 : 10 (I), 4 : 8 : 10 (II), 3 : 9 : 10 (III), 2 : 10 : 10 (IV), 1 : 11 : 10 (V), and 0 : 12 : 10 (VI) were used. Malaria and blood stains were better than the standard 5 : 7 : 10 (I) in III, IV and II in that order. Normal and leukemic human blood, mouse blood with Plasmodium berghei, and monkey blood with the CDC strain of Pl. falciparum were used as test materials. The staining mixtures were made from highly purified samples of azure B and methylene blue. Staining mixtures contained 12 ml 0.1% thiazin dye, 10 ml 0.1% eosin, 2 ml each of glycerol, methanol and 0.1 M phosphate buffer pH 6.5, 3 ml acetone as accelerator, and distilled water to make 40 ml; staining times of 10--30 min were used.


[Radiation therapy of carcinomas of the oral tongue and floor of mouth (experiences with 266 cases from 1950 to 1975) (author's transl)].

We report 266 patients, who were treated in the Universitätsklinik für Radiotherapie und Nuklearmedizin Zürich between 1950 and 1976. The relationship of the radiation dose to the tumor control and complication rate in various stages is analysed. In early stages, local control is achieved by a dose about 2200 ret. This dose is applicable without an undue complication rate by interstitial radiation with or without combination with percutaneous irradiation. The cure rate in this early stage is about 70 to 90% by irradiation. The cure rate in the stage T2N0 is about 50%, in T3N0 about 20% and in the other stages with cervical node metastases between 10 and 20%. In the majority of the locally advanced cases, a combined therapy with irradiation and surgery should be performed. In cases with multiple or fixed neck nodes, a combined therapy should be performed. In cases with a single neck node metastasis which is not too large, radiotherapy mostly is sufficient. In most cases with TxN0, prophylactic neck irradiation is indicated.


[A simple algorithm for practical calculations using the NSD concept (author's transl)].

An analytic equation is given, which allows simplified comparison calculations of treatment schedules in the validity range of the NSD concept. The equation is easily applicable, even in case of irregular fractionation, of treatment intermissions, and of changed schedules. The advantage of this unified calculation method is demonstrated by five examples, solving the problems occurring most frequently in practice.


[Own experiences with radiogold therapy of inoperable breast cancer in old patients (author's transl)].

Fifteen 62- to 85-year-old women with breast cancer are reported. An infiltration treatment by means of 198Au was performed with the patients because of general and local inoperability. Eleven of them were exposed to additional local X-irradiation. This therapy generally leads to local disappearance of the cancer disease.


The levels of immunoglobulins (IgG. IgA. IgM) under the effect of age and exposure to the mining environment in uranium industry.

IgG, IgA, and IgM levels were measured in 347 miners working in the underground of uranium mines; in 156 men entering into the work in mines, and in 101 men working in food industry and food distribution, all in the region of Central Bohemia. Significantly elevated IgG and IgM levels were found in the subgroup of miners exposed a short time (less than five years) to the mining environment, whereas elevated IgA levels were found in the subgroup exposed more than 15 years. The comparison of differences with age-matched controls has shown that the levels of IgM are significantly increased in the miners subgroup exposed to the mining environment no more than five years. In miners subgroups above 31 years a significant decline of IgG levels occurs. The expression of Ig levels' changes is not uniform in all exposed persons; an apparent heterogeneity occurs in miners population. Some differences in IgG and IgM levels were found between the two 31- to 40-year-old control subgroups. The possible conditions for differences found are discussed from the point of view of the effect of ionizing radiation and from the point of view of the stressing influence of environment and employment.


Liver polyribosome distribution intact and adrenalectomized rats exposed to gamma-radiation.

The mechanism(s) by which gamma radiation influences liver polyribosome distribution was studied in groups of intact and adrenalectomized male rats. The animals were divided among four subgroups: (1) control; (2) irradiation only; (3) cortisone only; (4) irradiation plus cortisone. Analysis of the liver polyribosome distribution patterns indicated that a shift from light to heavy aggregates occurred in both intact and adrenalectomized rats after they were exposed to gamma rays. In irradiated adrenalectomized rats, however, the shift to heavier aggregates was not as great as that which occurred in irradiated adrenal-intact animals. Subcutaneous injection of cortisone acetate (10 mg/100 g body weight) also altered the liver polyribosome patterns of both intact and adrenalectomized rats within 8 hours of its administration. The shift which occurred following cortisone administration, however, was less striking than that seen after irradiation only. Thus, although adrenal glucocorticoids contribute to the radiation induced shift in liver polyribosomes in adrenal intact rats, other factors appear to be involved, since the shift is also obtained in adrenalectomized animals.


Protective effect of a mixture of radioprotective substances (AET and mexamine) on the haemopoietic stem cells of mice.

A protective effect of a combined application of radioprotectors (AET -i.p., mexamine -s.c.) was studied in experiments on mice. Duration of the protective effect on the haemopoietic stem cells and on the survival of mice after a lethal exposure to X-ray radiation lasted for 5 hours after the injection. DRF calculated from a change of LD 50/30 reached the maximum (1.53) within 30 min after the injection, and was equal to 1.17 within an interval of 5 hrs. The protection of haemopoiesis, as expressed by DRF calculated from an equieffective exposure for 2 ESC, reached the maximum within a 60 min interval after the injection (DRF=3.4). D0 calculated from the CFU survival curves of endogenous haemopoietic tissue colonies (ESC) was, within a 120 min interval, almost three times higher (310 R) than in the control group (110 R). After the injection of radioprotectors a greater number of haemopoietic CFU survives at an exposure of 100-400 R than after mere irradiation. The postirradiation decrease in the weight of small intestine was smaller in protected animals than in the controls. The role of a decrease of the radiation damage to the haemopoietic stem cells in the total protective effect of the radioprotectors used is discussed.


Investigations on the modification of postirradiation pancreatic lipase activity by some endo- or exogenic factors. Part II. The influence of some ions, biogenic amines and hormones.

The subject of investigations was the influence of some endogenic substances: ions (K+ and Ca++), amines (serotonin, histamine and acetylcholine) and hormones (hydrocortisone and glucagon) on the lipase release from pancreatic granules (zymogen and lysosomes) of control and irradiated rats (800 R=24 hours). The decrease of enzyme release under the influence of hydrocortisone (both concentrations: 10(-6) and 10(-8) M), Ca++ ions, serotonin, histamine and acetylcholine after their administration in greater concentration (10(-6) M) was stated. Smaller concentrations of these substances evoked the transitory enhancement of enzyme activity. K+ ions and glucagon increased the lipase release (both concentrations). In the fraction of organelles from irradiated rats the action of K+ ions, glucagon and acetylcholine was diminished or abolished, while the inhibitory effectiveness (of greater concentrations) of Ca++ ions, serotonin, histamine and hydrocortisone was maintained.


Steroid hydroxylase induction in cultured human lymphocytes: effects of the menstrual cycle.

Steroid hydroxylases (SAH) are inducible in cultured human lymphocytes following treatment with estradiol-17beta. The enzyme systems induced are carbon monoxide sensitive and convert estradiol-17beta to a metabolite chromatographically indistinguishable from estriol. The level of inducibility of SAH varies drastically over a normal menstrual cycle with maximum induction in the late follicular phase and minimum induction during the luteal phase. The use of an oral contraceptive containing both a synthetic progestogen and ethynyl estradiol reduced SAH induction levels to those typically seen during the luteal phase of the menstrual cycle.


Thin-layer chromatographic in situ analysis of insect ecdysones via fluorescence-quenching.

The possibility of quantitating insect ecdysones in situ on thin-layer chromatographic plates was examined. Two approaches were evaluated: 1) the induction of ecdysone fluorescence by sulfuric acid treatments and 2) the fluorescence-quenching of fluorescent thin-layer chromatographic plates by ecdysones. The fluorescence-quenching method was found to be most suitable and had a linear response range from 0.5 to 3 microgram for alpha-ecdysone and 20-hydroxyecdysone. Fluorescence-quenching and high pressure liquid chromatographic analyses obtained from extracts of alpha-ecdysone 20-hydroxylase incubations gave similar results. New data concerning the acid-induced fluorescence of ecdysones showed alpha-ecdysone to be twice as fluorescent as 20-hydroxyecdysone.


Reactions of 4beta,5-epoxy-5beta-androstan-3-ones with hydrogen fluoride in pyridine.

4beta,5-Epoxy-5beta-androstane-3,17-dione (1a), 17beta-hydroxy-4beta,5-epoxy-5beta-androstan-3-one (1b) and 17beta-acetoxy-4beta,5-epoxy-5beta-androstan-3-one (1c) were treated with anhydrous hydrogen fluoride in pyridine (70% solution) at 55 detrees and yielded the corresponding 4-en-4-ols, e.g. 4-hydroxy-4-androstene-3,17-dione (2a). As the reaction temperature was lowered each epoxide formed a second product which, at -75 degrees, was the major component of the reaction mixture and was identified as the 5alpha-fluoro-4alpha-ol derivative of the parent enone, e.g. 4alpha-hydroxy-5-fluoro-5alpha-androstane-3,17-dione (3a). These fluorohydrins are thermally unstable, losing hydrogen fluoride. The acetates of -he fluorohydrins were also prepared, characterized, and shown to be more stable than the parent alcohols.


Progesterone receptor in the rat anterior pituitary: effect of estrogen priming and adrenalectomy.

The present study was done to determine if a progesterone receptor is present in rat pituitary. Cytosol was labeled with 3H-progesterone (3HP) or 3H-R5020 (3HR) and subjected to sucrose-glycerol density-gradient centrifugation. Serum progesterone was measured for correlation with progesterone receptor levels. Two 3HP-binding peaks (4S + 6S) were evident in uterine and pituitary cytosols. The 4S peak was eliminated by competition with unlabeled cortisol leaving a single 6S peak (progesterone receptor). Estradiol (E) priming of the male or female rat increased progesterone receptor levels in pituitary cytosol as demonstrated using 3HP and 3HR, and pituitary progesterone receptor bound 3HR with a higher affinity than 3HP. Following adrenalectomy of gonadectomized rats, progesterone receptor levels were increased in pituitary and uterine cytosol of both E-primed and unprimed groups. An inverse relationship was established between serum progesterone and progesterone receptor levels in the uterus and pituitary suggesting that stress-induced adrenal progesterone secretion significantly influences progesterone receptor levels in the rat. These results demonstrate an estrogen-inducible progesterone receptor in the rat pituitary with properties similar to those of the uterine progesterone receptor.


Polar corticosteroids in human neonatal urine; synthesis and gas chromatography-mass spectrometry of ring A reduced 6-hydroxylated corticosteroids.

This report describes the synthesis of 3alpha,6beta,11beta,17alpha,21-pentahydroxy-5beta-pregnane-20-one, 3alpha,6beta,11beta,17alpha,21-pentahydroxy-5beta-pregnane-20-one, 3alpha,6alpha,11beta,17alpha,21-pentahydroxy-5alpha-pregnane-20-one, 3alpha,6beta,17alpha,21-tetrahydroxy-5beta-pregnane-11,20-dione, 3alpha,6beta,17alpha,21-tetrahydroxy-5alpha-pregnane-11,20-dione, 3alpha,6alpha,17alpha,21-tetrahydroxy-5beta-pregnane-11,20-dione and 3alpha,6alpha,17alpha,21-tetrahydroxy-5alpha-pregnane-11,20-dione. The gas chromatographic-mass spectrometric properties of these compounds are given. Proof of structure was accomplished using gas chromatography-mass spectrometry, microchemical reactions, optical rotatory dispersion and nuclear magnetic resonance spectroscopy.


Preparation and characterization of (24R and 24S)-5beta-cholestane-3alpha,7alpha,24,25-tetrols and (24R and 24S)-5beta-cholestane-3alpha,24,25-triols.

(24R and 24S)-5beta-cholestane-3alpha,7alpha,24,25-tetrols were prepared by osmium tetroxide oxidation of 5beta-cholest-24-ene-3alpha,7alpha-diol. The resulting diastereomeric tetrols were separated by thin-layer chromatography, their purity ascertained by melting point, gas-liquid chromatography and mass spectra and their structural configurations were assigned by molecular rotation measurement and circular dichroism studies. In a similar fashion, the (24R and 24S)-5beta-cholestane-3alpha,24,25-triols were prepared and their structures identified.


Color reaction of cholesterol with trichloracetic acid and antimony trichloride. On the reaction mechanism.

The color reaction of cholesterol with trichloracetic acid and antimony trichloride was examined to elucidate its reaction mechanism. 3,5-Cholestadiene, 3,3'bis(3,5-cholestadiene), 3,3'bis(2,4-cholestadiene), and cholesteryl trichloroacetate were isolated as the reaction products from the colored reaction mixture of cholesterol, and the first three compounds were found to be responsible for the coloration. It was assumed that cholesterol was dehydrated to 3,5-cholestadiene and 2,4-cholestadiene, which were dimerized to 3,3'-bis(3,5-cholestadiene) and 3,3'-bis(2,4-cholestadiene), respectively, and 3,3'-bis(2,4-cholestadiene) was in part converted to 3,3'-bis(3,5-cholestadiene) in trichloroacetic acid and antimony trichloride. The free radicals were detected in the colored solutions of choelsterol, 3,5-cholestadiene, 3,3'-bis(3,5-cholestadiene), and 3,3'-bis(2,4-cholestadiene), and inferred to be the radical cations of the steroids. The radical cation was postulated to be responsible with respect to the mechanism of the coloration. The relationship between the color reagent and the formation of dimeric steroids was described.


Time-course relationships between serum LH, serum progesterone and urinary preganediol concentrations in normal women.

A specific and sensitive gas chromatographic method was used to investigate the concentration of pregnanediol glucuronide in urine in relation to the time of ovulation. Serum LH and progesterone concentrations in the same subjects were used as evidence for the occurrence of ovulation. The urinary concentration of pregnanediol glucuronide in 24-hour collections and in overnight specimens increased 2-fold or more from the day of the midcycle LH peak to the time of predicted ovulation (24-48 hour after the LH peak) in parallel with the rise in serum progesterone concentration.


A radioimmunoassay for danazol (17alpha-pregna-2,4-dien-20-yno[2,3-d]isoxazol-17-ol).

A method is described for the radioimmunoassay of circulating levels of the pituitary inhibiting agent, danazol. An antigen for danazol was prepared by reacting a 17-carboxy-methyloxime derivative of danazol with bovine serum albumin. By immunizing rabbits with this antigen, antiserum was generated which shows excellent specificity for danazol relative to its known metabolites as well as to many natural steroids. A radioimmunoassay was developed, without using separation or extraction techniques, involving competition for the antiserum between danazol in plasma and 14C-danazol. This assay has been successfully used to measure danazol in a series of normal human subjects receiving the drug at either 100 or 200 mg b.i.d. for 2 weeks. A significant relationship was seen between dosage of danazol and plasma concentrations.


Disulphates of 16-oxygenated ketonic C19 steroids as biliary metabolities of androsterone sulphate in female rats.

The chemical synthesis of 16beta-hydroxyandrosterone was described preparatory to studies of the disulphates of the 16-oxygenated ketonic C19 steroids present in the bile of female rats dosed with [3H]androsterone sulphate. The biliary metabolites were separated by chromatography on Sephadex LH-20 to afford monosulphate and diconjugate fractions. After solvolysis of the diconjugate fraction, the liberated steroids were separated by partition chromatography on Celite 545 and analyzed by gas chromatography-mass spectometry. In addition to 3alpha, 17beta-dihydroxy-5alpha-androstan-16-one isolated previously, 16beta-hydroxyandrosterone was identified as a disulphate.


Importance of jejunum for bile acid absorption in pigeons: effect of jejunal resection on plasma cholesterol and cholesterol excretion.

Previous studies from our laboratory suggested that the jejunum could be the major site of bile acid absorption in the pigeon. To confirm this observation directly, the effects of jejunal resection on the fecal excretion of bile acids and neutral sterols and on the level of plasma cholesterol were examined in the pigeon. This surgical procedure specifically increased (p less than 0.05) the excretion of bile acids, with no change in the excretion of neutral sterols. The total fecal steroid excretion in the pigeons with jejunal resection was significantly (P less than 0.01) higher than that in the control group. Jejunal resection also decreased the plasma cholesterol level in the pigeon. These results confirm our earlier suggestion that the jejunum is the major site of bile acid absorption in pigeons, unlike that in mammals, in which the ileum is the major site.


Testosterone metabolites in dog bile.

Testosterone-1,2-3H was injected intravenously into a male dog with a bile fistula and bile and urine collected. The radioactivity was excreted preponderantly in bile (52% of the injected dose) in 6 hours; only 12% appeared in the urine. Methods to study the biliary metabolites of testosterone in this and other animals were developed. Satisfactory conjugate patterns were obtained by fractionation on DEAE-Sephadex A-25 columns using two different elution systems. In addition to an unchanged fraction, six different monoglucuronide fractions were separated. No other conjugates were isolated. Lipidex 5000 column chromatography, TLC and paper chromatography were used for the isolation and purification of aglycone metabolites, which were further identified by co-crystallization methods. The biliary metabolites of testosterone were epiandrosterone (3beta-hydroxy-5alpha-androstan-17-one), etiocholanlone (3alpha-hydroxy-5beta-androstan-17-one), 5alpha-androstan-3beta, 17beta-diol, 5beta-androstan-3alpha, 17beta-diol and 5beta-androstan-3beta,17beta-diol.


Radioimmunoassay of 2-hydroxyestrone.

Under the protection of ascorbic acid a 2-hydroxyestrone bovine serum albumin conjugate was prepared containing intact 2-hydroxyestrone as determined by gas chromatographymass spectometry. Using this antigen highely specific antibodies were raised in rabbits. Cross-reactivity for 2-hydroxyestradiol and 2-hydroxyestriol was 26 and 4.5%, respectively. An assay procedure of 2-hydroxyestrone in human plasma is described. Using special precautions the assay allows the determination of 2-hydroxyestrone in plasma samples of women (50-95 pg/ml), pregnant women (105-220 pg/ml), men (45-65 pg/ml) and children(20-40 pg/ml).


The identification and quantification of three new 6alpha-hydroxylated corticosteroids in human neonatal urine.

The 24 hours urines of six two days old fullterm newborn infants were investigated for polar corticosteroids. 6alpha-hydroxy-tetrahydrocortisone, 6alpha-hydroxy-20alpha-cortolone and 6alpha-hydroxy-20beta-cortolone were identified by gas chromatographic-mass spectrometric comparison of the urinary steroids to compounds synthesized previously. These 6alpha-hydroxylated corticosteroids as well as seven other polar corticosteroids were quantified by gas chromatography or mass fragmentography. It was shown that the newly identified steroids constituted a quantitatively important part of the neonatal urinary corticosteroids. The unconjugated- and glucuronic acid conjugated steroids were quantified separately. It was found that the extent of glucuronoconjugation decreased with increasing polarity of the steroid moiety.


Uptake and metabolism of androgens by bovine spermatozoa.

Spermatozoa from bovine ejaculates and cauda epiditymidis were incubated with either tritiated 17 beta-hydroxy-5 alpha-androstane-3-one (DHT) or 5 alpha-androstane-3 alpha, 17 beta-diol (3 alpha-diol). Examination of the medium incubations demonstrated metabolic conversion of both DHT and 3 alpha-diol when these steriods were incubated with ejaculated sperm. In addition to this interconversion, the following metabolities were identified: 5 alpha-androstane-3 beta, 17 beta-diol, (3 beta-diol), androsterone and 5 alpha-androstane-3, 17-dione (5 alpha-A-dione). Incubations with cauda spermatozoa showed similar metabolic patterns. Androgen binding was exhibited by both sperm types. Examination of the washed cauda sperm pellet, following incubations with 3 alpha-diol showed that the incubated steroid was the most abundantly bound. DHT and 5 alpha-androst-16-en-3 alpha-ol (delta 16-3 alpha-ol1 were also detected. The major part of the radioactivity bound in the sperm pellet was identified as DHT when this steroid was used as the substrate; the remaining radioactivity consisted of 3 alpha-diol and delta 16-3 alpha-ol. Investigations of ejaculated sperm pellets gave similar results apart from the additional identification of 5 alpha-androst-16-en-3 one (delta 16-3-one) and 5 alpha-androst-16-en-3 beta-ol (delta 16-3 beta-ol (delta 16-3 beta-ol).


Identification of bile alcohols in normal rabbit bile.

Rabbit bile was examined for bile alcohols. Using combined gas chromatography-mass spectrometry, 5 beta-cholestane-3 alpha, 7 alpha, 12 alpha, 25,26-pentol, 5 beta-cholestane-3 alpha, 7 alpha, 12 alpha, 25--tetrol, 5 beta-cholestane-3 alpha, 7 alpha, 12 alpha,24 beta-tetrol, 24-methyl-25-homo-5 beta-cholane-3 alpha, 7 alpha, 12 alpha, 24-tetrol, and 5 alpha-cholestane-3 alpha, 7 alpha, 12 alpha, 24 beta-tetrol were identified as the minor constituents of normal rabbit bile.


Identifications of 5 beta-cholestane-3 alpha, 7 alpha, 12 alpha, 23 beta-tetrol, 5 beta-cholestane-3 alpha, 7 alpha, 12 alpha, 24 alpha-tetrol, and 5 beta-cholestane-3 alpha, 7 alpha, 12 alpha, 24 beta-tetrol in cerebrotendinous xanthomatosis.

The bile alcohols present in the feces of a patient with cerebrotendinous xanthomatosis were studied. Three bile alcohols which are different from any known natural bile alcohol were isolated as minor components of the fecal bile alcohol fraction. The structures of these compounds were established as 5 beta-cholestane-3 alpha, 7 alpha, 12 alpha, 23 beta-tetrol, 5 beta-cholestane-3 alpha, 7 alpha, 12 alpha, 24 alpha-tetrol, and 5 beta-cholestane-3 alpha, 7 alpha, 12 alpha, 24 beta-tetrol by comparison with synthetic samples.


The presence in bovine trachealis muscle of cytosol receptors with high affinity for glucocorticoids.

Whereas receptors with high affinity for glucocorticoids have been reported present in both fetal and adult lungs, there has been no attempt to identify such receptors in airways smooth muscle. We have examined the binding characteristics of glucocorticoids for a cytosol fraction prepared from bovine trachealis muscle. The cytosol fraction, which was prepared in 10 mM Tris, 1.5 mM Na2 EDTA and 2 mM dithiothreitol, contained macromolecular high affinity receptors for 3H-dexamethasone (Kd = 5.6 nM) at a concentration of 0.20 pmol/mg protein. The comparative affinities of other steroids in descending order were: beclomethasone-17, 21-dipropionate, dexamethasone, beta-methasone, triamcinolone and triamcinolone acetonide. Progesterone and deoxycorticosterone only bound to a limited extent, whereas the binding with 5 alpha-dihydrotesterone and pregnenolone was negligible. We conclude that airways smooth muscle is a target tissue for glucocorticoids and postulate that the binding of glucocorticoids to these receptors may receptors may represent the initial step in the anti-asthmatic action of these drugs.


A simple and efficient method for raising steroid antibodies in rabbits.

The production of estradiol antibodies by two immunization procedures was monitored by means of an enzyme-immunoassay. Procedure A consisted of three intramuscular injections given at two-week intervals, followed by five intravenous booster injections and procedure B consisted of multiple intradermal injections given once. Procedure A gave much higher antibody titer. In both procedures the sensitivities of assays using the antisera increased initially and reached a plateau after three to four months of immunization. Consistent changes in specificities were observed. A shortened procedure A is proposed as a simple and efficient procedure for raising steroid antibodies in rabbits.


Various 18-substituted progesterones.

In order to test the potential biological activity of 18-substituted progesterones, 3,20-dioxo-4-pregene-18-carbonitrile (ld approximately) was converted to 3,20-dioxo-4-pregnene-18-carboxylic acid (lb approximately) and methyl 3,20-dioxo-4-pregnene-18-carboxylate (ld approximately) via a sequence of reactions involving an intramolecular hydrolysis of the 18--arbonitrile. Lithium aluminum hydride reduction of the bisethylene ketal derived from la approximately furnished 18-aminomethyl-5-pregnene-3,20-dione 3,20-bisethylene ketal (8 approximately). Acetylation and hydrolysis furnished 18-acetamidomethyl-4-pregnene-3,20-dione (lf approximately) and simple hydrolysis of 8 approximately furnished 3'alpha H-5' 6'-dihydro-2',19 beta-dimethyl-3-oxo-4-goneno [13,17-c]pyridine (9 approximately). None of the compounds exhibited any activity in Clauberg or anti-Clauberg tests.


A radioimmunoassay method for urinary catechol estrogens.

A radioimmunoassay method for urinary catechol estrogens is described; The specific nature of the antisera allows direct analyses of acid hydrolyzed urine. A LH-20 Sephadex column chromatography can be employed for individual determinations of 2-hydroxyestrone and 2-hydroxyestradiol. The excretion of catechol estrogens during menstrual cycles ranged from 14.48 to 50.15 microgram per 24 hours, whereas, during the last trimester of pregnancies, the values ranged from 129.30 to 1758. 20 microgram per 24 hours.


An exchange assay for the cytoplasmic glucocorticoid receptor in the liver of the rat.

Cytosolic receptor for glucocorticoids can exist in either the free or bound form; assays now in use measure only the free form. In order to assay the total glucocorticoid receptor content of rat liver, free plus bound, we have developed an exchange assay wherein specifically bound [3H]dexamethasone is shown to be a valid measure of receptor in the presence of high concentrations of corticosterone. The exchange between [3H]dexamethasone and corticosterone is able to proceed because, unter the conditions of the assay, corticosterone is almost completely metabolized.


The extrasplanchnic origin of blood dihydrotestosterone in elderly men.

The splanchnic extraction and interconversion of testosterone (T) and dihydrotestosterone (DHT) were studied in 5 elderly men undergoing cardiac catheterization using a constant infusion of [1,2-3H] testosterone and [4-14C] DHT. Metabolic clearance rate (MCR), splanchnic extraction (SE), splanchnic clearance (SC), extrasplanchnic clearance (ESC), transfer constant in blood ([p] T-DHT BB) and transfer constant across the liver ([p] T-DHT SB) were calculated. The MCRT was 675 +/- 108 (mean +/- SC) L/day and MCRDHT was 409 +/- 68 L/day. SET was 45.9 +/- 7.0% and SEDHT was 18.5 +/- 5.4%. When these values are compared with those recently reported by us for normal men, there is a 1/3 reduction in SET and 1/2 reduction for SEDHT in elderly men. The calculated SCT and ESCT were 355 +/- 72 L/day and 320 +/- 86 L/day, respectively. SCDHT and ESCDHT were 145 +/- 48 L/day and 263 +/- 77 L/day respectively, suggesting that a major fraction of DHT is metabolized in extrasplanchnic organs. No evidence for a net appearance of DHT by either mass or specific activity analysis in hepatic vein blood was observed indicating that the splanchnic compartment does not contribute DHT into the circulation either by de novo synthesis or via conversion from testosterone. This work indicates that conversion of testosterone to DHT in elderly men occurs entirely in extrasplanchnic tissue.


A new and improved synthesis of 19-iodocholesterol 3-acetate.

19-Iodocholesterol 3-acetate (VI) was synthesized in a single step by iodo group substitution for hydroxyl using either one of two different reagents: (1) carbodiimidonium methiodide (VIII) or (2) triphenyl-phosphine/N-iodosuccinimide (IX). The yields were as satisfactory as those obtained from the two step iodide replacement of a 19-hydroxy group via the 19-tosyloxy group. The principal intermediate, 19-hydroxy cholesterol 3-acetate (V), was derived in appreciable quantities, and relatively inexpensively, through the Pb (OCOCH3) 4 photolytic oxidation of the bromohydrin of cholesterol 3-acetate (III) to the epoxide (IV) thence Zn reduction to the 19-hydroxy compound. A specially designed 12 liter flask was of aid in accomplishing the photolysis reaction. Dry column chromatography with the supportive puncture sampling was integral to achieving the good yields and high purity of 19-iodocholesterol (VIII).


Metabolism of diethylstilbestrol in the C3H mouse: chromatographic systems for the quantitative analysis of DES metabolic products.

Initial excretion studies with orally administered [monoethyl-1-3H] DES demonstrated the feces to be the principal mode of elimination of DES in the C3H mouse. Metabolic studies with tritiated DES and/or [UL-14C] DES were performed with orally dosed C3H high (MTV+) and low (MTV-) titer MMTV female mice. Extraction and partitioning of the fecal radioactivity demonstrated 77 to 86% (n = 4) to be benzene soluble and the remainder H2O soluble. The principal product in the organic phase following Sephadex LH-20 and HPLC purification was DES. The aqueous phase was resolved by LH-20 into two conjugate fractions that were partially hydrolyzed by beta-glucuronidase. The principal aglycone was chromatographically identical with authentic DES. The urinary conjugates were resolved into six fractions. The four major fractions were 80% hydrolyzable with beta-glucuronidase. Two of these fractions had trans-DES as the principal aglycone, whereas the other two had a major peak similar to but not chromatographically coincident with cis-DES. In certain experiments mice were sequentially dosed with tritium (24 hr) followed by a 14C dose (24 hr). Two mice (MTV+) were also previously fed 1000 ppb DES prior to these experiments. The tritated and 14C products were combined and analyzed simultaneously. This experiment did not reveal significant differences in the metabolism due to the modes of radioactive labeling, MMTV titer, or the prior feeding of DES. The developed methodology was judged to purify quantitatively 90% or more of the DES radioactive products.


5alpha-Reductase and 3alpha-hydroxysteroid oxidoreductase enzyme activities in epididymis and their control by androgen and the rete testis fluid.

The 5alpha-reductase and 3alpha-hydroxysteroid oxidoreductase enzyme activities have been measured in epididymal tissues and the control of these activities by androgens and the rete testis fluid appreciated. The highest 5alpha-reductase enzyme activity was found in the caput, the lowest in the corpus epididymidis. Androgens have a positive control on the 5alpha-reductase but no effect on the 3alpha-hydroxysteroid oxidoreductase activity. Ligation of the efferent ducts decreased significantly both enzyme activities in the caput but not in the corpus or in the cauda epididymidis.


An improved synthesis of 6beta-hydroxy-5beta-pregnane-3,20-dione.

6beta-Hydroxy-5beta-pregnane-3,20-dione, formerly prepared by hydroboration method, has been obtained in greatly improved yield by a simpler irradiation-hydrogenation procedure.


Identification of 7xi-hydroxytetrahydrocortisol and 7xi-hydroxytetrahydrocortisone from urine of newborn baboons (Papio papio).

Two highly polar, unconjugated metabolltes of tetrahydrocortisol and tetrahydrocortisone, which had been shown previously to account for greater than 40% of [14C]cortisol (F) administered to baboon neonates, were isolated from urine of a term newborn animal. Metabolites were extracted with ethyl acetate, purified by sequential paper chromatography and crystallization, and identified by chemical and biochemical procedures. Both metabolites of F are 5beta-pregnanes with a hydroxyl of unknown orientation at C-7, a 3alpha-hydroxyl, and a dihydroxyacetone side chain. The two compounds differ from each other by the presence of either oxo or hydroxyl functions at C-11. The following nomenclature is proposed: unknown 1: 3alpha,7xi,11beta,17,21-pentahydroxy-5beta-pregnan-20-one (7xi-hydroxytetrahydrocortisol); unknown 11: 3alpha,7xi,17,21-tetrahydroxy-5beta-pregnane-11,20-dione (7xi-hydroxytetrahydrocortisone). Production of these compounds compensates quantitatively for the decreased formation of F glucuronoside metabolites by baboon neonates, compared with that in adults. Therefore, we propose that 7-hydroxylation is a major alternative pathway for F metabolism in the perinatal period of the baboon.


Studies of the effect of 5alpha-androstane-3alpha,17alpha-diol in the canine prostate in vivo.

Adult beagle dogs, castrated one month previously, were treated with 5alpha-androstane-3alpha,17alpha-diol (total dose 300 mg) over a period of one month. Examination of the prostate after treatment showed no significant change in size, weight or histological appearance from the castrate dog prostate. Subsequent administration of 5alpha-androstane-3alpha,17beta-diol (300 mg) over the same period of time resulted in restoration of the prostate size, weight and histological appearance to that of the normal intact dog prostate. It is concluded that exogenously administered 5alpha-androstane-3alpha,17alpha-diol does not promote prostatic growth in the castrate beagle dog.


A general procedure for estimation of corticosteroid response in individual rats.

A procedure is presented whereby microliter volumes of plasma, obtained rapidly and repeatedly from the tail vein of unanesthetized rats, can be analyzed for corticosterone. Preliminary results suggest that (1) basal levels are considerably lower and (2) the latency to corticosteroid stress response is shorter than previously reported.


Stabilization of placental aromatase by dithiothreitol in the presence of oxidizing agents.

Cumene hydroperoxide, sodium periodate and iodosobenzene were not able to support aromatization by placental microsomes in the absence of NADPH or molecular oxygen. In the presence of these oxidizing agents and NADPH, aromatase was slowly inactivated. Dithiothreitol (10mM) prevented the loss of aromatizing activity in the presence of these compounds. One function of dithiothreitol may be to protect aromatase by scavenging harmful oxidizing agents.


21-Hydroxypregna-1,4-diene-3,11,20-trione: a biliary metabolite of a cartilaginous fish, Raja sp.

21-Hydroxypregna-1,4-diene-3,11,20-trione was isolated from skate bile and as an in vivo metabolite of 3H-1alpha-hydroxycorticosterone. Identity was established by chromatography and derivatization to constant 3H/14C ratio and mass spectrometry of the 20,21-acetonide. The new steroid was present in the free form and as the glucuronoside.


Participation of propionate in cholesterol biosynthesis by rat liver.

Incorporation of (214C) propionate into cholesterol was demonstrated using rat liver slices and homogenates. The incorporation of (214C) propionate was greater than that of (214C) acetate. Using the same liver homogenate preparation (214C) succinate and (214C) pyruvate were incorporated into cholesterol to a lesser extent than (214C) acetate and (214C) propionate. Addition of unlabeled acetate failed to dilute the incorporation of (214C) propionate. Incorporation of the 2 and 3 carbon atoms pf propionate were equal; little incorporation of the 1 carbon atom was demonstrable. These results indicate that propionate is an excellent source of 2 carbon units for isoprenoid biosynthesis; the intermediary pathway does not involve a common acetate pool nor can these results be satisfactorily explained by citric acid intermediary metabolism.