[The effects of testosterone and estradiol on the activity of lysosomal enzymes in rat liver and kidneys]. In in vitro tests with lysosomes isolated from the liver and kidneys of castrated rats of both sexes the action of testosterone and beta-estradiol in concentrations of 3.76.10(-4)M on the activity of beta-glucosidase, beta-galactosidase and acid phosphatase was investigated. Testosterone is shown to reduce the total and free activity of the membrane-bound enzymes and to increase the release from the matrix lysosomes. Estradiol proved less active than is testosteron. The renal lysosomes in vitro are more sensitive to the action of sex hormones than are hepatic lysosomes. In the interaction of testosterone and estradiol with lysosomal membranes a sex specificity was revealed.


Inhibition of ovulation in marmoset monkeys by indomethacin. The effect of indomethacin on human menopausal gonadotropin/human chorionic gonadotropin-induced ovulation was studied in marmoset monkeys. Indomethacin was effective in preventing follicular rupture and ovum extrusion when administered simultaneously with gonadotropin. The production of progesterone and luteinization of the granulosa cells were not affected by indomethacin treatment. Histologic examination of corpora lutea from indomethacin-treated animals revealed the presence of entrapped oocytes surrounded by luteal cells. These results suggest that prostaglandin synthesis is required for ovulation in this species, but not for luteinization and progesterone production.


Clinical and experimental aspects of the anti-mammary carinogenic activity of estriol. Intermittent implantation of 600--1,300 microgram estriol subcutaneously beginning 48 h before oral administration of 7,12-dimethylbenz(a)anthracene or procarbazine prevents development of 80--90% of carcinomas of the breast occurring during the natural life span of the intact female Sprague-Dawley rat. Some estriol precursors were less inhibitory of breast cancer development among 23 other estrogens and androgens, progestins and glucocorticoids tested. More frequent or lower estriol doses than 100--200 microgram/kg/24 h every 2 months were less inhibitory of breast carcinogenesis. No other types of neoplasms were reduced in incidence by estriol implants, which also reduced uterine weights by 20--25%. Intermittent substitution of estriol for estrone or estradiol in the nuclear receptor complexes of target cells probably accounts for these observations, which resemble the effect of castration in reducing breast cancer incidence. Human studies indicate excellent tolerance for oral estriol doses of 10--200 microgram/kg/24 h, which may correct subnormal estriol/estrone + estradiol urinary quotients associated with elevated risk of breast carcinogenesis in epidemiologic investigations.


[Studies on the activities of glycogen synthetase and glycogen phosphorylase in the human endometrium (author's transl)]. Glycogen content, glycogen synthetase and glycogen phosphorylase were studied in the endometrial tissues of 28 women with normal menstrual cycles and in the decidual tissues of 24 women with normal early pregnancies. The endometrial glycogen synthetase enzyme increased gradually from the proliferative phase to the secretory phase and reached maximal activity during the sixteenth to twenty-third days of the cycle, a time coincident with maximal glycogen content, while the glycogen phosphorylase reached its maximal activity on and after the twenty-fourth day of the cycle. In addition, glycogen phosphorylase activity in the decidual tissue during the early period of gestation (4--10 weeks) was lower than that in the endometrial tissue during the late secretory phase of the cycle, and in particular, the active form (-AMP) was significantly (p less than 0.005) low, with the result that the glycogen content in the decidua was significantly (p less than 0.05) higher than that in the endometrium. On the other hand, in 20 normal pregnant women during the 6--10th weeks of gestation, the glycogen contents in the decidua and in the placental villi were 599 +/- 44 mg/100 g wet weight (mean +/- standard error of the mean) and 731 +/- 55 mg/100 g, respectively, but the difference between them was statistically not significant. The levels of glycogen synthetase and glycogen phosphorylase enzyme in the decidua were significantly (p less than 0.005) higher than those in the placental villi.


[Demonstration of immunoreactive GIF-like substance in villi and decidua by radioimmunoassay and immunofluorescence (author's transl)]. Since Arimura et al (1975) reported the radioimmunoassay for somatostatin (GIF), the concentration of GIF in various organ and brain regions were determined by radio-immunoassay. Dubois et al (1975) reported that immunohistochemically somatostatin was located in the discrete cells of the pancreas as well as the hypothalamus, and from this result, they presented the concept of local hormone instead of systemic hormone which was up to that time accepted in endocrinology. In this study, we developed the high specific anti-GIF serum using rabbits, and with the micro immunodiffusion method, we demonstrated that the precipitin band formed a circular fusion between the GIF and anti-GIF serum. This pattern of reaction was also seen in decidual immunodiffusion. In addition, we developed the radioimmunoassay for GIF using this anti-serum and measured immunoreactive GIF-like substances in villi and decidua of early pregnancy. The concentration of GIF-like substances with 2 N acetic acid extracted of villi and decidua were 0 to 30 pg/0.1 g dry weight. At the same time, we demonstrated the presence of GIF-like substance-containing cells in the villi and decidua by indirect immunofluorescent method. The intensity of immunofluorescence was in cytotrophoblast rather than syncytiotrophoblast, and decidual stromal cell also reacted to the immunofluorescence.


Growth hormone response to arginine infusion in malnourished children. Pituitary response to stimulation was assessed by intravenous arginine infusion in 17 malnourished children and 7 controls. Following stimulation, serum growth hormone levels increased from 6.6 +/- 1.2 to 14.9 +/- 2.9 ng/ml in the marasmic group and similarly from 7.3 +/- 1.8 to 11.9 +/- 2.7 ng/ml in the controls. By contrast, in kwashiorkor the mean growth concentration was high and remained unchanged (19.6 +/- 3.9 ng/ml before and 17.2 +/- 2.1 ng/ml after stimulation). No difference was observed between groups for serum insulin concentration. These results do not lend support to the suggestion that there might be atrophy or pituitary hypofunction in infantile malnutrition.


Competitive social strategies in groups of deprived and experienced rhesus monkeys. Behavior during competition for water was observed in 2 social groups of young rhesus monkeys (3 females, 3 males in each). Monkeys in one group were socially deprived and those in the other were socially experienced (raised with mother and agemates). Social status, based on dyadic recording of displacements at the water bottle, was predictive of a number of measures related to water consumption and social orientation in both groups, but this measure was less reliable and predictive for the experienced group than for the deprived groups, but this measure was less reliable and predictive for the experienced group than for the deprived group. A major reason for the comparatively low predictive value and reliability of status among experienced monkeys was their ability to influence the behavior of higher status members through responses directed to a 3rd party and other elaborate social strategies, many of which depended on responding to status relations between 2nd and 3rd parties. The fact that such strategies were only observed in the experienced group is a clear indication that the development of higher orders of social cognition is dependent on early social experience.


The neurological manifestations of chronic inhalation of leaded gasoline. Abnormal neurological signs were found in 46 of 50 children and adolescents chronically sniffing leaded gasoline. These abnormalities resolved within eight weeks in all but one case. Exaggerated deep reflexes, postural tremor and evidence of cerebellar dysfunction occurred in a highly significnat number of patients. Forty-nine had blood lead levels greater than or equal to 40ug/dl. The mean blood lead levels were significantly higher in those with (a) abnormally brisk deep reflexes and (b) with evidence of cerebellar dysfunction, than in those without these findings. Five optional treatment regimes were employed and a classification was used, based on clinical findings, initial blood lead levels and the response to the calcium disodium edatate mobilization test. 39 patients received chelation therapy. These data suggest that neurological manifestations occur frequently in those abusing leaded gasoline and that chelation therapy has an important place in their management.


Changes of the cell cycle during carcinogenesis of hamster cheek pouch epithelium by 7,12-dimethylbenz[a]anthracene. Changes of the cell cycle during carcinogenesis of hamster cheek pouch epithelium were studied and it was demonstrated that in hyperplastic and neoplastic stages, the cell cycle time decreased to approximately 2/3 and 1/3 of the normal control, respectively. A nominal growth fraction was found to exceed unity in carcinogen-treated epithelium, whether hyperplastic or neoplastic. This result seems to indicate that a considerable number of cells in which DNA synthesis is stimulated by the carcinogen fail to enter mitosis. A possible implication of repair synthesis is also discussed.


Susceptibility of human cultured cells to Mason-Pfizer monkey virus. Mason-Pfizer monkey virus (M-PMV), originally isolated from mammary carcinoma of a rhesus monkey, is infectious to various human cells in tissue culture. Human epithelioid cells (HeLa and HEp-2), fibroblastic cells (embryonic cells as well as virus-transformed embryonic RSb cells), and glial cells (KC) have all been infected with M-PMV. Electron microscope studies have shown that all infected cells produce progeny virus. The titer of infectious M-PMV in cultures of HeLa and HEp-2 cells assayed by KC syncytium formation was 10(1) syncytium-forming unit (SFU) per 0.2 ml. The titers for whole human embryonic cells (HEC), human embryonic lung cells (HEL), and embryonic skin Detroit-551 cells were 10(4) SFU/0.2 ml for each, and 10(3) SFU/0.2 ml for RSb and KC cells.


Gallstone duodenum: unusual complication of an obstructing intraluminal duodenal diverticulum. An unusual case of gallstones within the duodenum, secondary to obstruction by an intraluminal diverticulum, is reported. The gallbladder appeared normal an oral cholecystogram, excluding internal biliary fistula as the source of duodenal gallstones.


Angiographic demonstration of non-occlusive hepatic infarction with scintigraphic and microscopic correlation. The angiographic appearances of histologically proven hepatic infarction without vascular occlusion has not, to our knowledge, been previously described. A case of non-occlusive hepatic infarction in a diabetic patient is presented and correlated with scintigraphic and microscopic findings. It would appear to be the first recorded case of hepatic infarction associated with diabetic ketoacidosis. A follow-up angiogram showed resolution. The literature is briefly reviewed.


Bacillus cereus-induced malabsorption in young mice. Following a single, oral dose of Bacillus cereus (2 X 10(8) bacteria) in vitro intestinal absorption of D-glucose, D-galactose, L-arginine, L-histidine, L-ornithine and L-proline in young mice (aged 2--3 1/2 months) decreased. Malabsorption of D-glucose was dose- and time-dependent. Impaired absorption of D-glucose occurred throughtout the length of the small intestine, particularly distally. Following hydrolysis of D-maltose at the brush border, D-glucose absorption in infected mice and that of the untreated controls was similar. Using D-glucose, fluid transfer in the infected intestine and that of the controls was alike. Although slightly lower, fluid transfer in the infected intestine using the other solutes was not significantly different compared with the controls. Glucose-dependent and glucose-independent intestinal fluid transfer in infected animals was like that of the controls. Using old infected mice (aged 8--9 months) intestinal absorption of D-glucose and L-histidine was unchanged compared with young mice. The fresh small intestinal weight in infected mice and the controls was alike. Changes in the histology of the small intestine in young infected mice were small and inconsistent.


Costs of gastrointestinal examinations: a comparative study. The costs of performing four major gastrointestinal examinations were studied and compared: single-contrast and double-contrast upper gastrointestinal series, and single-contrast and double contrast barium enema. A  total of 1,725 examination records were reviewed for the number of films expended, room occupancy time, and physician involvement time, and the costs of the studies determined. The data indicated a large and significantly increased time and expense required for the double-contrast barium enema over the single-contrast barium enema. A relatively small increment was found in the cost performing the double-contrast upper gastrointestinal series over the single-contrast study.


Clinical and histopathological evolution of active chronic hepatitis. The clinical and histopathological evolution of 7 children with severe chronic active hepatitis and 8 with moderate chronic active hepatitis were studied. The majority (11-15) of the children had a clear past history of acute viral hepatitis. The cases of chronic active hepatitis with a moderate activity had a favorable evolution, but not the cases of severe chronic active hepatitis. Four of them developed into liver cirrhosis, in two the morphological alteration did not improve, and only one case showed a chronic presistent hepatitis.


[Action of an anti-viral on the Australia antigen in patients with hepatitis A virus and in healthy carriers]. Patients with positive serum HBAg reactions were treated with Viramid (1-beta-D-ribofuranozyl-1, 2, 4-triazol-3-carboxamide) a new anti-viral drug. Of the patients involved in this study, 5 had chronic hepatitis or prolonged hepatitis and 12 were healthy HBAg carriers. Results obtained suggest that the drug had some action on serum HBAg titres with a decrease or negativation of HBAg serum titres in peripheric blood, the evaluation of which was immediately seen by employing the counterimmune electrophoresis technique.


[Liver abscess. Diagnosis by the method of opacification]. A 15 min infusion of a 75% iodized solution at 2 cc/k-w produces opacification of the more vascularized organs; hypovascular, necrotic or cystic regions are thus of a different radiological density. In order to show the usefulness of this method in diagnosing a hepatic abscess quickly and topographically, it was applied in 13 cases which had been diagnosed as invading amoebiasis. Hepatography proved to be the most valuable. Positive findings are observed 5 to 7 min after infusion has been completed. The greater density at the edge of the abscess enables its diameter to be measured and the liquid content to be calculated fairly accurately.


Hemostasis in schistosomiasis mansoni.

Hemostasis in schistosomiasis mansoni. Hemostasis was studied in 18 patients with the hepato-intestinal form of Schistosomiasis mansoni and in 23 with the hepato-splenic form. Both groups are compared referring to alterations found. The relations between plasmatic coagulation factors and hepatocytic function tests are studied. In 6 patients the explorations were repeated after administration of hycanthone. In the patients with the hepato-splenic form, the presence of chronic consumptive coagulophathy was found. The coagulopathy disappeared either by treatment with heparine or by splenectomy. This observation points out the importance of splenomegaly in the development of chronic consumptive coagulopathy, and may be an important factor in indicating splenectomy in a patient with portal hypertension due to Schistosomiasis mansoni.


Chronic treatment with azathioprine and cyclophosphamide in rats: structural and functional effects on small intestine and liver. A report is presented of the effects upon the liver and enteric mucosa of rats treated chronically with azathioprine (AZA) and cyclophosphamide (CY) in equiactive doses three times greater. The groups treated either with equiactive doses of AZA and CY or with doses three times greater of AZA and CY presented limited and moderate morphological alteration in the liver and in the mucosa of the small intestine, while all rats treated with high dose of CY died in the first phase of the experiment.


[Incidence of hepatitis surface antigen among all the blood donors of the blood banks of the Venezuelan Institute of Social Security in the federal district]. The presence of H Bs-Ag was studied by means of counterimmunoelectrophoresis in the sera of 879 voluntary donors who gave blood between May and September 1974 at the federal District's blood banks of the Venezuelan Institute of Social Security. Eight hundred twenty-eight were male and fifty one female. Five sera samples were positive HBs-Ag which represents an incidence of 0.56%. A bibliographic review of the subject is made. The necessity of making mandatory the determination of HBs-Ag in all blood prior to transfusion is emphasized.


The intra-hepatic bile ducts in hepatic steatosis and in congested livers. The authors study 25 steatotic or congested livers by means of post-mortem cholangiographies and vinyl moulds of the bile ducts. They show that the bile ducts of these organs present alterations, characterized by stretching, rectification, and lengthening of the intra-hepatic bile ducts, as a consequence of the diffuse hepatomegaly that occurs in the disease. They relate the average number of straight ducts in the intra-hepatic bile tract to the extension of the extension of the disease, obtaining a positive relation. A statistical evaluation of the results was made, allowing objective conclusions.


Post-mortem aeroportography.

Post-mortem aeroportography. The authors suggest a new methodology for the study of the intrahepatic portal system, using atmospheric air as a contrast medium. They used 48 livers of normal and pathological corpses, obtaining post-mortem cholangiographies and vinyl molds. Comparing the images obtained with literature data and with the vinyl molds, the authors conclude that the method is accurate, presenting furthermore many advantages with regard to contrasts generally used, such as low cost, easy performance, possibility repeating the radiographs as many times as necessary, and the possibility of simultaneous obtaining contrasts of other intra-hepatic systems by use of other radio-opaque contrast media in another system, as for example the biliary or the arterial system.


[Treatment of acute pancreatitis by glucagon. Experimental studies in dogs]. The authors made a comparative study of the use of glucagon (4 mg/day IV) in 30 dogs with acute pancreatitis. They found that the mortality and amylasemia were significantly lower in the glucagon-treated group than in the control groups (saline and glucose solution 5%). In the glucagon group, the areas of necrosis were smaller (1.5 cm) and rarely found; microscopically, the areas of necrosis and the inflammatory reaction were much smaller than in the other groups. These findings lead to the conclusion that the beneficial action of glucagon is due to another mechanism other than its hyperglycemic effect and that the administration of hypertonic solutions of glucose does not have a beneficial effect in acute pancreatitis.


[Pancreatic fistula].

[Pancreatic fistula]. The authors present the result of an investigation on the use of cement and bentonite in the treatment of pancreatic fistulas that coexisted with permeable duct and which has been closed by the local application of said elements.


[Comparative radiographic study of the stomach using technics of double contrast]. One hundred patients with gastroduodenal symptoms were studied: 50 at a general hospital and 50 at a private hospital. The first group was studied with the Japonese method recommended by Heizaburo Ishikawa and used in Venezuela with some modifications by Zaidman. The second group was studied with the technique developed by Valencia-Parparcén. In addition, in both groups 4.5 mg of an anticholinergic drug (Hsp-2986 Merck-Darmstad-Germany) was applied intravenously to obtain gastroduodenal hypotony. The results from both groups showed that double contrast X-rays are better than the standard and are important factors in the judgment of the radiological diagnosis of gastroduodenal diseases. The methods do not exclude, but rather complement, each other, and are important contributions when a radiography in series with dosified compression through fluoroscopy cannot be carried out under optimum conditions.


[Toluidine blue and gastric cancer].

[Toluidine blue and gastric cancer]. The authors used Toluidine Blue to stain gastric lesions in nine patients. Six cases were found with adenocarcinoma that stained an intense blue that was limited to the extension of the malignancy itself. The other three cases did not stain and their histological diagnosis had been benign chronic gastric ulcers. One patient did color faintly due to atypical cells in the ulcer crater. The authors concluded that it is a procedure easy to apply, simple, harmless, and inexpensive, that gives results and permit the differentiation of malignant lesions from benign ones. They therefore recommend the method to gastroenterologists, endoscopists, and surgeons to guide and direct the taking of biopsies, specially in small cases, incipient or multiple stomach lesions.


[Internal hemorrhoidectomy by ligation]. The present work consists in the treatment of 60 patient suffering from internal hemorrhoids in the first, second and third degree, with and without complications. We utilized a new semi-surgical methods that is based on the ligation of these with satisfactory results in 85% of the cases and an improvement in 9.33%.


[Cancer of the gastric stump in gastrectomies for benign diseases]. The authors present 8 cases of cancer in the gastric stump of patients operated for benign pathologies. They reviewed 800 clinical histories of gastric cancers with microscopic confirmation. Incidence is of 1%. They analyzed age, sex, interval of time elapsed since the first operation, diagnosis, treatment and mortality of these patients. They emphasize that diagnosis is difficult and confusing but must be considered in patients with gastrectomies for benign causes done long ago and that present new symptoms or aggravation of the preexisting ones. For this reason these patients must be carefully worked up and studied. The mortality is very high. Treatment is surgical and radical.


[Superselective vagotomy of parietal cells. Evaluation and results in duodenal ulcer]. The authors present a study carried out jointly by the Gastroenterology Service and Surgical Service No. 2 of the General Hospital of the Venezuela Institute of Social Security "Dr. Ildemaro Salas". The results and objectives of the operation are analyzed insisting on its lower morbility and mortality, as well as on the lower incidence of diarrhea, dumping and other gastrointestinales discomforts. The first operation was performed on 17 november 1971 and up to the present 74 patients have been operated, 65 for duodenal ulcers, 7 for esophageal hernia of the hiatus, 1 case of hemorrhagic gastritis, and 1 case of prepyloric ulcer. Some details are given of the operations, the indications for it are pointed out, specially gastric hyperacidity, symptomatic ulcer, and absence of duodenal stenosis. There were 7 post-operative complications and 2 cases of mortality. As to the results there are presented 92% successful interventions and 8% failures.


[Intraluminal diverticulum of the duodenum. Report of a case and review of published cases]. The intraluminal diverticulum of the duodenum is very rare; it is an anatomical sac like formation, located in the interior of the duodenum and communicating with it through an orifice situated a the upper pole of the diverticulum where the wall is covered by mucosa on both faces. It has interesting etiopathogenic problems. We present an observation after reviewing the corresponding literature. It was a 32-year old woman suffering from moderate epigastric pains, pyrosis, chronic constipation, markedly nervous. The epigastric pain was unrelated to food or physical efforts. The physical examination showed slight epigastric pain. The gastroduodenal radiological examination showed in the second part of the duodenum a particular image, pear-shaped, homogeneous, impregnated in barium; this image was surrounded by a radiotransparent halo which changed with the patient's position. It has been seen that the diverticulum changes in size and position in agreement with the peristaltism, without ever obstructing the passage through the duodenum. A review is made of the literature and history of intraluminal diverticulum.


[Arterio-mesenteric occlusion of the duodenum in Chagas' disease]. The authors present eight cases of arterio-mesenteric duodenal ileus diagnosed in the Serviço de Cirurgia de Ribeirão Preto (São Paula--Brasil) among 2.573 surgical interventions of digestive tract performed in 15 years. Seven (87.5%) cases presented positive complement fixation test for Chagas' disease and signs of Chagas' cardiopathy and/or enteropathy. Comparing similar casuist studies published in Brazil, the authors suggest that American Trypanosomiasis must be considered as a contributive factor in the genesis of arterio-mesenteric duodenal ileus in Latin America.


[Study of the length of a stay of partial gastroduodenectomized patients]. The length of stay for 482 partial gastrectomy patients in the Servico de Cirurgia de Ribeirao Preto in the period from 1960 to 1974 is compared with that of four other neighbor hospitals for the year 1973. There was reduction in the length of stay during these years and it was more obvious when compared the other four hospitals. The authors claim that early discharge and home management should always be recommended and suggest an analysis of factors interfering with length of stay.


[Comparative study of cicatrization with suture technics in 1 or 2 layers in small intestine anastomosis in adverse conditions. Experimental work in dogs]. Some experimental procedures have shown in intestinal anastomosis advantages in the use of suture on only one plane as compared to the classical suture on two planes. However, these experiments, performed in healthy dogs, raised doubts about its safety when used under adverse clinical conditions. The objective of this work is, therefore, to compare in dogs submitted to immunosupressive drugs (azotioprine and corticoid) the two techniques of suture in the presence of peritonitis and intestinal ischemia. The comparative study of the healing in the sutures performed under such conditions shows the advantages in the use of suture on only one plane. In the dogs examined on the fourth and fifteenth day after surgery, the superiority of the suture on one plane compared to that on two planes is evident, both in the macroscopic and microscopic aspects. The suture in one plane shows a better coadaptation of the borders, without increasing the thickness of the wall at the level of the anastomosis, less exudative inflammatory reaction, quicker and greater fibroblastic proliferation, less inflammatory reaction to the suture material, and less vascular alteration at the healing level.


[Treatment of injuries of the duodenum and pancreas]. The authors study 84 patients that had damages of the duodenum and or pancreas and underwent different types of surgery. In 39 patients who had damage only in the duodenum; there were 2 (4.0%) duodenal fistulas, and 3 (6.0%) intraperitoneal abscess. In 38 patients who had injuries only in the pancreas; there were 6 (13.1%) acute pancreatities, pancreatic fistulas and 1  (2.2%) pancreatic pseudocyst. In 7 cases, injuries were found to both pancreas and duodenum. The author reports 1 case of duodenal fistula (14.3%) 1 case of acute pancreatitis (14.3%) and 3 cases of pancreatic fistula (43.0%). Only 7 patients died (8.3%), and of these 2 died for reasons not directly related to the operatory technic used. A simple suture can be performed in those cases where a complete section of the duodenum is unnecessary and there is no injury to the duodenal papilla. A burying suture of the stomas associated with a side-to-side duodenojejunal anastomosis should be preferred in the complete section of the duodenum localized beyond the ampulla of Vater. In all superficial wounds of small extension a suture of the pancreas should be performed. Distal pancreatectomy of body or tail should be made in the wounds with a possible lesion to the Wirsung duct and when there is an extensive glandular lesion. The associate wounds of duodenum and pancreas should be treated as if they were isolated lesions.


Duhamel procedure for sigmoid cancer.

Duhamel procedure for sigmoid cancer. A description is given of the advantages of the Duhamel technique over that carried out by colorectal anastomosis at the level of the pelvis. The anastomosis is performed at the level of the perineum. Ten patients with cancer of the colon were operated and it was determined that there was no tumoral relapse of the residual ampulla. There were no complications and this technique can be used as a substitute for Hartmann's.


[Stenosis and intestinal ulcer due to chronic ingestion of a combination of a thiazidic diuretic with potassium]. Reports from recent years shown evidence of the existence of a type of primary ulceration of the small bowel associated with stenosis, relatively frequent and possibly iatrogenic. The primary characteristics of the studied cases were that it was present in patients over 50 years old, a good number of them had cardiorenal diseases or were hypertensives, and ingestion of a thiazide diuretic supplemented with potassium. Clinical and experimental investigations suggest that potassium, with or without a diuretic, could cause ulceration and stenosis of the small bowel. Presented in this report the case of a 62-year-old patient with cardiovascular disease and chronic ingestion of a thiazide diuretic with potassium chloride (Diclotride-K (R) who developed an intestinal obstruction that merited surgical intervention which showed a circumscribed stenosis of jejunum accompanied by ulceration.


[Changes of serum proteins in schistosomiasis mansoni]. The total and fractional proteins were determined in 174 out-patients from a rural area that had schistosomiasis mansoni and had not received specific treatment, and in 51 subjects also from another rural area and therfore having similar epidemiological characteristics but not in the endemic area of bilharziasis. In schistosomiasis patients there was a notable change in the fractional proteins, the hypoalbuminemia and hypergammaglobulinemia being significant irrespective of the clinical forms of the disease. The intermediate fraction between the beta and gamma globulins, described by some authors, was not found, possibly due to the fact that the serums analyzed were from out-patients with the less serious clinical forms. In the group from the non-endemic area of bilharziasis hypoalbuminemia was the most significant alteration found.


[Etiopathogenical and clinical aspects of chronic pancreatitis in San Pablo (Brazil): observations of 80 cases]. The etiopathogeny and clinical aspects of chronic pancreatitis were studied in 80 patients. From the etiological point of view it was observed that 75 out of the 80 patients (93,75%) had had previous intense alcohol ingestion (mean 239.95 +/- 158.34 g of pure ethanol/day) during an average period of 12.5 years. A study of their diet showed that these patients ate significantly more protein than the controls (p less than 0.001). In 2 of the remaining 5 patients, the familiar form of the disease was characterized; in 1 there was a previous history of biliar disease and no etiological factor could be detected. The pathogenic implications of the excessive consumption of alochol associated with a high protein diet, as well as the main clinical characteristics presented by these patients are discussed.


[Intramural hematoma of the digestive tract in relation to anticoagulant therapy]. Three cases of submucosas hematomas of the gastrointestinal tract are presented in association with anticoagulant therapy. The clinical and radiological criteria for their diagnosis is discussed.


[Gastrinemia and basal gastric acid secretion]. A group of 43 subjects was studied: 22 normal, 6 with gastric ulcers, and 15 with duodenal ulcers. A correlation was made between gastrinemia and basal gastric acid secretion. The findings suggest that the basal gastric secretion does not play an important role in either the basal gastrinemia in normal subjects or in those with gastric ulcers. The correlation was significant and negative between basal gastric secretion and basal gastrinemia in those with duodenal ulcers, which might express differences in the self-regulation of gastrinemia related to antral pH.


[Strongyloidiasic hyperinfestation].

[Strongyloidiasic hyperinfestation]. A study was made of twelve cases of fatal Strongylodiasis with Hyperinfestation which presented a severe picture of the small intestine and colon, some with localization in the lungs, bronchus, and other organs. Preliminary results are given of a prospective study related to the bacterial flora and its role in the pictures of larvarial aggression, which could lead to greater knowledge about the biology of the parasite, and therefore to interesting findings of a therapeutic nature.


[Role of the gastroenterologist in the fate of the functional gastrointestinal patient. Comparative evaluation in 2 series of patients in the outpatient clinic of the Social Security Gastroenterology department of Barquisimeto]. A comparative evaluation is made of patients with functional digestive diseases in an out-patient clinic of the Social Security of Barquisimeto. In the first group of patients a routine approach was followed while in the second group a brief biographic history is added adapting it to the circumstances of work that exist within this type of clinic. From the information obtained of the destiny of the patients of each group, it is evident that the total approach to the patient is of importance as the only mean possible for an adequate approximation to the patient and the proper management of the case. The problem is presented of the gastroenterologist, overwhelmed by the great technical progress, confronted by the destiny of the functional digestive patient in socialized medicine.


[Portal hypertension by hepatic cirrhosis. Selective decompression by distal renal spleno anastomosis. Report of a case]. An analysis is made of the new techniques used in the surgical treatment of portal hypertensión, specially the selective descompression of the gastroesophageal varices. A case is presented of surgery performed in the Surgical Service IV of the Hospital Universitario de Caracas using the technique described by Warren.


Genetic analysis of two allelic temperature-sensitive mutants of Drosophila melanogaster both of which are zygotic and maternal-effect lethals. After fertilization, the development of a zygote depends upon both gene products synthesized by its maternal parent and gene products synthesized by the zygote itself. To analyze genetically the relative contributions of these two sources of gene products, several laboratories have been isolating two classes of mutants of Drosophila melanogaster: maternal-effect lethals and zygotic lethals. This report concerns the analysis of two temperature-sensitive mutants, OX736hs and PC025hs, which were isolated as alleles of a small-disc mutant, l(3)1902. These alleles are not only zygotic lethals, but also maternal-effect lethals. They have temperature-sensitive periods during larval life and during oogenesis. Mutant larvae exposed continuously to restrictive temperature have small discs. One-or two-day exposures to the restrictive temperature administered during the third larval instar lead to a homeotic transformation of the midlegs and hindlegs to the pattern characteristic of the forelegs. Mutant females exposed to the restrictive temperature during oogenesis produce eggs that can develop until gastrulation, but do not hatch.--The existence of these mutants, and one that was recently described by another group, implies that there may be a class of genes, heretofore unrecognized, whose products are synthesized during oogenesis, are essential for embryogenesis and are also synthesized during larval stages within imaginal disc cells.


[New derivative of nitromidazole-tinidazole, for the treatment of giardiasis]. The author treated 100 patients infected by Giardia lamblia with a new antiprotozoal agent: Tinidazole. The patients were divided in two groups: Group I, composed of 50 patients were divided with a dosage of 300 mgm per day for seven consecutive days. The author obtained a cure rate of 96% (48 cases), and minimal side-effects were observed in 10% of the cases: nausea and headache, Group II also composed of 50 patients which were treated with a single dose of 2,0 gm. The parasitological cure obtained was around 90% (45 patients) and the tolerance to the drug was very good. To evaluate the cure, the methods of Faust et al. and Hoffman, Pons & Janer, such as spontaneous sedimentation in water, were used, 7, 14 and 21 días after treatment. The author concluded that the new antiprotozoal agent is an excellent drug against Giardia lamblia infections.


Genetic and biochemical basis of enzyme activity variation in natural populations. I. Alcohol dehydrogenase in Drosophila melanogaster. Recent studies by various authors suggest that variation in gene regulation may be common in nature, and might be of great evolutionary consequence; but the ascertainment of variation in gene regulation has proven to be a difficult problem. In this study, we explore this problem by measuring alcohol dehydrogenase (ADH) activity in Drosophila melanogaster strains homozygous for various combinations of given second and third chromosomes sampled from a natural population. The structural locus (Adh) coding for ADH is on the second chromosome. The results show that: (1) there are genes, other than Adh, that affect the levels of ADH activity; (2) at least some of these "regulatory" genes are located on the third chromosome, and thus are not adjacent to the Adh locus; (3) variation exists in natural populations for such regulatory genes; (4) the effect of these regulatory genes varies as they interact with different second chromosomes; (5) third chromosomes with high-activity genes are either partially or completely dominant over chromosomes with low-activity genes; (6) the effects of the regulatory genes are pervasive throughout development; and (7) the third chromosome genes regulate the levels of ADH activity by affecting the number of ADH molecules in the flies. The results are consistent with the view that the evolution of regulatory genes may play an important role in adaptation.


Experimental evolution of a new enzymatic function. II. Evolution of multiple functions for ebg enzyme in E. coli. The evolution of ebgo enzyme of Escherichia coli, an enzyme which is unable to hydrolyze lactose, lactulose, lactobionate, or galactose-arabinoside effectively, has been directed in successive steps so that the evolved enzyme is able to hydrolyze these galactosides effectively. I show that in order for a strain of E. coli with a lacZ deletion to evolve the ability to use lactobionate as a carbon source, a series of mutations must occur in the ebg genes, and that these mutations must be selected in a particular order. The ordered series of mutations constitutes an obligatory evolutionary pathway for the acquisition of a new function for ebgo enzyme. A comparison of newly evolved strains with parental strains shows that when ebg enzyme acquires a new function, its old functions often suffer; but that in several cases old functions are either unaffected or are improved. I conclude that divergence of functions catalyzed by an enzyme need not require gene duplication.


[Gastric candidiasis. Report of a case]. One patient was seen with Candida albicans infection in a gastric ulcer. Gastric biopsy was of value in the diagnosis. The patient responded well to Nistatin therapy.


[Effect of EcoRI restrictase on the transforming DNA of different bacilli]. The transforming activity of DNA from Bacillus subtilis 168, Bac. subtilis W23, Bac. subtilis NRS and Bac. aterrimus after the EcoRI restrictase treatment was studied. The auxotrophic strains of Bac. subtilis 168 were used as recipients in bacterial transformation. The transforming activity for different markers decreased up to 0.001--6 per cent from the initial level for different Bacilli DNAs. In some cases the sensitivity of the same marker from different Bacilli differed in more than 50 times. Bac. subtilis and Bac. aterrimus have demonstrated the maximal differences. Such differences can be used for the identification of close related Bacilli.


[One of the classes of revertants of a rec H Bacillus subtilis mutant]. A class of revertants of Bacillus subtilis mutant rec H, which completely restored the ability to transformation but without restoring the activity of ATP-dependent deoxyribonuclease, is isolated and studied. Reversions are located in the same chromosome region as the original mutation. The detection of such revertants points out the existence of more than one recombination pathway for Bac. subtilis transformation.


[Nature of the mutations that determine the ability of Bacillus subtilis A-50 to sporulate at high glucose concentrations in the medium]. The ability of Bacillus subtilis A-50 to sporulate in the medium containing high glucose concentrations is caused by at least two mutation types: pts mutations and cat (or tgl) mutations, both of them affecting differently the level of alkaline proteinase synthesis. The decrease of the level of enzyme activity in the case of pts mutation (gluR3 mutant) occurs at the expense of glucose transport disturbance. The mutation cat (tgl) (mutant gluR5) causes the increase in enzyme synthesis at the expense of catabolic resistance to glucose of genes controlling alkaline proteinase synthesis and the spore formation in Bac. subtilis A-50. cat5(gluR5) and pts3(gluR3) mutations are located on the chromosome of Bac. subtilis in the region metD and argC respectively. The over-synthesis of alkaline proteinase characteristic of Bac. subtilis A-50 is controlled by the polygenic system, as the level of alkaline proteinase synthesis in argA+ transformants makes up 25% of the level of activity of the original strain. The productivity of Bac. subtilis A-50 can be enhanced by introducing an additional cat mutation.


[Effect of repeated administration of glucocorticoid on the glycogen level of mouse muscle (author's transl)]. Glycogen content and glycogen synthetase activity in mouse skeletal muscle (M. gastrocnemius) after single or repeated administrations of glucocorticoid were investigated and compared with those in liver, in a series of experiments to elucidate the biochemical mechanism of steroid myopathy. Results obtained were as follows. After a single administration of triamcinolone acetonide (TA, 5 mg/kg i.p.), glycogen content in muscle increased maximally at about 12 hr and returned to control level at about 24 hr. The I form activity of glycogen synthetase in muscle was increased only 6 hr after a single administration, while the total activity of the enzyme was not significantly changed. Similar results were obtained with the liver. After repeated administrations of TA for one week, the glycogen accumulation in the muscle was lowered markedly as compared with that after a single administration. No increase in synthetase I activity was observed, or rather, the enzyme activity was decreased as compared with that from control. Similar results were obtained with the liver. After single or repeated administrations, blood glucose level decreased slightly, glucose level of muscle was not affected and G-6-P level of muscle increased markedly. These results suggest that the lower accumulation of glycogen in the muscle after repeated administrations of glucocorticoid may be primarily due to an abnormality in the glycogen synthetase system.


Non-transformable mutants of Bacillus subtilis defective in the penetration of DNA into the cell. Four non-transformable mutants of Bacillus subtilis 168 defective in the penetration of DNA into the recipient cell were isolated. All mutants were fully non-transformable with mutation in genes influencing irreversible binding of the donor DNA by the recipient cell.


Lactase activity of microorganisms.

Lactase activity of microorganisms. Sixty-two strains of yeasts, molds and bacteria were screened for lactase (beta-D-galactosidase) activity. Strains exhibiting the enzyme activity were evaluated for cell yield as well as enzyme units available per litre of the medium, per g cell dry weight and per mg protein of their cell-free extracts. The molds exhibited lowest enzyme activity but highest cell yields, bacteria produced lowest cell yield and maximum enzyme activity. Cultures exhibiting very high activity among yeasts were Saccharomyces fragilis (strain 3217) and among bacteria Streptococcus cremoris (strain H), Lactobacillus bulgaricus (strain RTS and 1373) and Leuconostoc citrovorum (strain 8081).


Specific endonucleases in Streptomyces aureofaciens. Two strains of Streptomyces aureofaciens were found to contain restriction endodeoxynucleases; S. aureofaciens IKA 18/4 contains Sau I which splits lambda DNA into three fragments, S. aureofaciens IKA 22201 Sau Ii which splits lambda DNA into more than 15 fragments.


Food sharing among captive Leontopithecus rosalia. Captive L. rosalia were observed to exchange food routinely in family groups, a process varying from donation to theft. Donation involved the use of 'invitational' signals which operated most efficiently in parent-infant sharing and in donating live prey. Food moved, by sharing and theft, toward: (a) males prior to the female's first conception; (b) females during late pregnancy; (c) carriers of infants; (d) weanlings; (e) juveniles when infants died; (f) third animals in trios, regardless of their age and relatedness to the pair. Successful reproduction, a complex behavior involving all group members in cooperative infant care and provisioning, is probably enhanced by captive L. rosalia's food-sharing habits.


Diurnal behavior of Lemur mongoz at Ampijoroa, Madagascar. Lemur mongoz was observed for 28 h between February and July in Ampijoroa, northwestern Madagascar. The animals were diurnal, with peaks of activity in mid-morning and late afternoon. This finding contrasts with other reports that L. mongoz at the same site and somewhat later in the year are nocturnal. Group size was three or four. L. mongoz associated with L. fulvus during group progression and feeding and when giving alarm responses to man. Vocal signals consist of a range of intergrading grunts and shrieks; olfactory signals of apparent scent marking with the anogenital region, forehead, and palms of the hands; visual signals of mainly unritualized body movements and postures. All of these signals are quite similar to signals of L. fulvus.


Cytogenetics and taxonomy of some South Bolivian monkeys. Illustrations are given by the authors of chromosome banding patterns of three species of monkeys: douroucouli, black-capped capuchin and squirrel monkey. The precise locality of the animals in southern Bolivia is known. This enables a taxonomic discussion on the value of some names of these animals.


The social behavior of a marmoset (Saguinus fuscicollis) group III. Spatial analysis of social structure. The social behavior, and particularly the spacing patterns, of a marmoset (Saguinus fuscicollis) group in a semi-naturalistic enclosure were observed for 14 months. Data analysis revealed various changes as the group grew in size from four to eventually six members. Weekly mean distances between the adult pair supported a spatial measure for estrus for the female. Group dispersion, mean squared distance between all possible pairs, seemed to vary with the age composition of the group. Factor analysis of location correlation matrices by 4-week blocks resulted in age-related subgroupings. Each of the adult pair formed an independent subgroup except for behavioral estrus and early infant care periods when they formed one subgroup. The offspring, initially attached to the adults, gradually moved into juvenile/subadult subgroups. Increasing spatial independence was shown as an animal approached adulthood at 24 months of age.


[Neurofibroma and neurofibrosarcoma in Recklinghausen's neurofibromatosis]. The neurofibromatosis von Recklinghausen is a systemic disease, which can cause various different changes in the body. The case report of a female patient shows all the typical signs of neurofibromatosis von Recklinghausen. These signs are café-au-lait spots, congenital tibia pseudarthrosis, abdominal plexiform neurofibromatosis and peripherical neurofibromas with sarcomatous degeneration.


Pituitary responsiveness to thyrotropin releasing hormone in insulin-dependent diabetes mellitus. The pituitary responses to the intravenous administration of 200 mg of Thyrotropin Releasing Hormone were investigated in 14 poorly controlled insulin dependent diabetic males and in nine matched controls. The mean TSH and prolactin responses in the two groups were similar although both tended to be lower in the diabetics. There was a small FSH rise in 11 of the 23 subjects.


The use of intramuscularly administered methyl-TRH to evaluate the pituitary-thyroid axis. The present study was carried out to evaluate the effectiveness of intramuscular administration of methyl-TRH, a potent analogue of thyrotropin-releasing hormone, for assessing pituitary reserve of TSH and prolactin and for distinguishing euthyroid, hypothyroid and hyperthyroid individuals. Serum samples were taken for 24 hours after intramuscular injection of methyl-TRH, 200 microgram, in 19 euthyroid subjects, 9 hypothyroid men and 9 hyperthyroid men. The mean serum prolactin and TSH concentrations were significantly elevated over baseline levels at 30 min in the euthyroid individuals and remained elevated for 3 to 4 hours. The serum TSH, T3 and T4 responses after intramuscular methyl-TRH in euthyroid subjects were clearly distinguishable from those of hyperthyroid and hypothyroid patients. Significant elevation of the serum T3 and T4 concentrations at 24 hours after intramuscular injection of methyl-TRH shows the sustained effect of this TRH analogue in euthyroid subjects.


[Vitamin B 6 biosynthesis in Bacillus subtilis]. 1) A vitamin-B6-producing mutant, BA 1, was selected by treatment of Bacillus subtilis with N-methyl-N'-nitro-N-nitrosoguanidine. Using gradient plates supplemented with the vitamin B6 antagonist isonicotinohydrazide, three mutants of BA 1 were isolated, which excrete 2-5 mg of vitamin B6/l of growth medium. 2) Mutation of the three vitamin-B6-producing strains BA 1, BA 11 and L 71 led to the isolation of 49 vitamin-B6 deficient mutants. All mutants are able to grow with pyridoxine, pyridoxal, pyridoxamine, and even with 4'-deoxypyridoxine. Glycolaldehyde or nicotinic acid do not support growth of the mutants. Some of these vitamin-B6-deficient mutants can also grow in the absence of vitamin B6, providing isoleucine is present. Others show a growth stimulation, when isoleucine is added to a medium containing a vitamin B6 compound. Isoleucine can be replaced by 3-methyl-2-oxovalerate. Cross-feeding experiments indicated a division of the mutants into two groups. Using chromatographic methods, substances which support growth of the mutants were purified, but have not yet been identified. Following the addition of 4'-deoxypyridoxine, 4'-deoxypyridoxine 5'-phosphate was isolated from the growth medium of a vitamin B6-deficient mutant. 3) Threonine dehydratase, transaminase B and transaminase C from wild-type Bacillus subtilis were compared with the enzymes from vitamin-B6-producing strains and with the enzymes from vitamin-B6-deficient mutants. Both the vitamin-B6-producing and the vitamin B6-deficient mutants show higher specific activities than wild type. In the mutant strains no multivalent repression of the threonine dehydratase and transminase B by isoleucine, leucine and valine could be demonstrated. Leucine dehydrogenase, the first enzyme of the isoleucine catabolic pathway, is constitutively produced in the vitamin-B6-producing and in the vitamin-B6-deficient mutants. In the vitamin-B6-deficient mutants, there is a correlation between growth yield in the presence of isoleucine and the specific activity of leucine dehydrogenase. In the crude extract of Bacillus subtilis no pyridoxamine-phosphate oxidase activity could be demonstrated, whereas pyridoxal kinase was readily detectable.


Human plasma kallikrein: purification, enzyme characterization and quantitative determination in plasma. Human plasma kallikrein was isolated from a plasma fraction related to Cohn fraction IV4 by affinity- and Sephadex G-150 chromotography yielding a material with 17.3 TAME-U/A280 unit. The preparation was characterized by immunological and enzymatic methods. Complex formation with alpha2-macroglobulin, C1-inactivator and aprotinin was demonstrated by immunoelectrophoresis. The bradykinin release from high-molecular weight kininogen and its inhibition by antibodies to kallikrein, AT III and AT III-heparin complex were measured using a biological test system (rat uterus). Time dependent inactivation of kallikrein by AT III, and AT III-heparin complex was shown by means of a synthetic kallikrein substrate: Bz-Pro-Phe-Arg-pNan. The same substrate was used to measure the activation of prekallikrein in plasma by kaolin and F XII a. Antibodies raised against kallikrein were shown to inhibit the reaction specifically. A quantitative determination of plasma prekallikrein by electroimmunodiffusion according to Laurell was developed: the plasma concentration in normal individuals was found to be 1.8 - 2.2 TAME-U/ml related to kallikrein activity; this corresponds approximately to 9 - 11 mg antigen/100 ml plasma.


The modification of the lone tryptophan residue in human serum albumin by 2-hydroxy-5-nitrobenzyl bromide. Characterization of the modified protein and the binding of L-tryptophan and benzodiazepines to the tryptophan-modified albumin. The possible function of the lone tryptophan residue of human serum albumin in the stereospecific binding site for indole and benzodiazepine compounds was investigated by chemical modification. This residue can be selectively modified with 2-hydroxy-5-nitrobenzyl bromide. The modification alters the conformation of the albumin only slightly, as revealed by circular dichroism, fluorescence, and ultraviolet absorption measurements. A decrease in the association constants of L-tryptophan and diazepam of about 30 - 50% and a decrease in the extrinsic Cotton effects of four benzodiazepine derivatives of about 10 - 15% were found as specific effects of the tryptophan modification. The tryptophan modification itself did not change the number of binding sites of diazepam and L-tryptophan. It is suggested that the lone tryptophan residue of human serum albumin is not directly involved in the specific binding site for indole and benzodiazepine compounds. However, the modification alters the properties of the binding site either by an incomplete refolding of the albumin after urea treatment, or a more selective allosteric effect of the modified tryptophan residue.


Day hospitalization as a cost-effective alternative to inpatient care: a pilot study. Two private day hospitals and an insurance company offering group health coverage to federal employees undertook a pilot study to determine if providing insurance coverage for day hospitalization on the same basis as for inpatient treatment was a feasible means of controlling the cost of psychiatric care. The study included 31 patients who otherwise would have been hospitalized; most had histories of severe psychiatric disorders and extensive treatment. Using the measure that the day hospital patients would have been in inpatient treatment for the same number of days, the authors estimate that the use of day treatment saved the insurer more than $255,000. They recommend that day hospitalization be reimbursed on the same basis as inpatient care if a day hospital can meet stringent criteria ensuring that it provides active, appropriate treatment, and they present a list of such criteria.


Clinical affiliations needed to augment educational experience. Too little attention has been paid to the need for a solid clinical component in the health professional's education program. So that this important component is added to the didactic component, hospitals should work with educational institutions as coequal partners to arrive at workable affiliation agreements.


Deciphering the price tag on government regulations. Studies show that about one and a half to two percent of total hospial costs is spent on complying with regulations. But, some observers say the costs are even higher.


[Serum levels and urinary excretion of zinc, magnesium, calcium and phosphates during postoperative parenteral feeding]. In 45 patients serum levels and urinary excretion of zinc, magnesium, calcium and phosphate were examined during postoperative parenteral nutrition. The patients received the same parenteral nutrition and were divided in groups according to different types of operations. Serum levels of calcium and magnesium were found to be normal in all groups  at highly varying degrees of urinary excretion. Zinc and phosphate serum levels, as well as urinary excretion, show a markedly different behavior.


[Parenteral feeding after ureterosigmoidostomy: nitron balance, free plasma amino acids and state of proteins]. The postoperative protein metabolism of 11 patients who had required either a colon conduit or ureterosigmoidostomy urinary diversion was examined. From the 3rd to 10th postoperative day day, each patient received a standard parenteral alimentation (100 g amino acids and 2000 kcal/24 h). Nitrogen balance, plasma level of the free amino acids, and the C3c-, C4-complement and transferrin fraction in plasma were determined regularly. The results showed that this substitution therapy succeeded in avoiding serious alterations in the parameters measured.


[General aspects concerning the intravenous feeding of cancer patients]. In order to obtain or maintain a good nutritional status in cancer patients, it is often necessary to perform intravenous nutrition. In summary, several studies have indicated that intravenous nutrition may be beneficial in association with surgery, radiation, or chemotherapy in patients with cancer. More controlled studies, however, are required. There is no indication at the present time of any adverse effects of this method of treatment in relation to tumor growth. The general nutritional improvement in patients on intravenous nutrition increases the immunocompetence, resistance to radiation and cytostatic as well as the mood and quality of life of the cancer patients. In very broad terms this new intravenous nutrition therapy means that a cancer patient should not be left without specific cancer therapy because of starvation and its serious or even fatal complications.


Immunogenic capacity of ribosomes of Salmonella typhi interfered with a flagellin-like material contaminant. The double-immunodiffusion technique and sodium dodecyl sulfate-polyacrylamide electrophoresis were used to demonstrate the presence of flagellin-like material strongly attached to ribosomes of Salmonella typhi Ty 2. This flagellin-like material contaminating the ribosome preparation interferes with the induction of antiribosome serum promoting the formation of antisera reacting either only with flagellin or in some cases with flagellin and ribosomes, but giving a very weak reaction with the latter. The interference is also observed when purified ribosomes from a nonflagellated mutant of S. typhi (S. typhi O-901) mixed with purified S. typhi Ty 2 flagellin are utilized as antigens. The antiribosome sera obtained with ribosomes from S. typhi O-901 have a considerably higher titer than those that are interfered with. These sera were able to react with ribosomes obtained from several related species and did not react with flagella-derived flagellin of S. typhi Ty 2.


Isolation of the L-phase variant from toxigenic Corynebacterium diphtheriae C7(beta). The L-phase variant was isolated from lysogenic strain C7(beta) of Corynebacterium diphtheriae by the penicillin disk tecnnique on an agar medium containing 3.7% brain heart infusion, 3% NaCl, 1.25% Grycine, 1% Noble agar, and 10% horse serum. The L-phase formed a "fried egg" colony; it was resistant to antibiotics that inhibit biosynthesis of bacterial cell walls but was susceptible to antibiotics that act on the protein-synthesizing system. After several transfers of the L-phase initially isolated, the L-phase of strain C7(beta) became stable, forming a typical L-phase colony on the agar even without penicillin. The L-phase variant retained lysogenicity and toxigenicity. The toxin production by this L-phase variant was, like that by the bacterial phase, inhibited by excess iron.


Effect of immunosuppression on Rickettsia rickettsii infection in guinea pigs. The role of the immune response in the pathogenesis of Rickettsia rickettsii infection in guinea pigs was investigated by immunosuppression, using antilymphocyte serum. Twenty guinea pigs were inoculated with R. rickettsii, Sheila Smith strain, on day 0. Fifteen animals received antilymphocyte serum on days --1, 0, 2, 4, and 6. Five animals received normal rabbit serum on the same schedule. At necropsy, specimens were collected for histological examination, rickettsial immunofluorescence, rickettsial titration, and antirickettsial antibody titration. All normal rabbit serum recipients and 12 of 15 antilymphocyte serum recipients developed typical disease. Comparison of animals in terminal stages of disease revealed the same clinical course and gross lesions, but differing rickettsial burden and cellular response. Immunosuppressed animals had higher titers of splenic rickettsiae and greater numbers of immunofluorescent rickettsiae. Thus, although antibody was undetectable in both groups, there appeared to be an inhibition of antirickettsial immunity. Microscopic vasculitis was similar quantitatively, but differed qualitatively, with immunocompetent animals having the typical monouclear/lymphocytic inflammation and immunosuppressed animals having neutrophilic predominance. This study demonstrates that immunopathological mechanisms are not necessary for the pathogenesis of experimental Rocky Mountain spotted fever. The rickettsiae themselves seem capable of causing cellular and tissue damage.


Experimental infection of the genital tract of female grivet monkeys by Mycoplasma hominis. Mycoplasma hominis, a common inhabitant of the mucosae of the genitourinary tract of human and nonhuman primates, was inoculated directly into the uterine tubes of five laparotomized grivet monkeys. A self-limiting acute salpingitis and parametritis developed within a few days in all animals. Although there were no clinical signs of overt disease, the gross pathology was characterized by pronounced oedematous swelling and hyperaemia of the tubes and parametria. Microscopically, cellular infiltrations of lymphocytes and some polymorphonuclear leukocytes were found in the acute phase in the subserosa and muscularis of the tubes and in the parametria. Granulation tissue and fat necrosis appeared at a later stage in the parametria. The infection was associated with a marked antibody response and a moderate rise of the erythrocyte sedimentation rate and leukocyte counts. The capability of M. hominis to produce salpingitis and parametritis in a nonhuman primate would seem to add rather significantly to the available evidence suggesting an etiological role of this organism in inflammatory diseases of the internal female genitals of humans.


Impaired host resistance to endotoxin and malaria in polychlorinated biphenyl- and hexachlorobenzene-treated mice. The in vivo effect of polychlorinated biphenyl (PCB) and hexachlorobenzene (HCB) on murine endotoxin sensitivity and resistance to malaria (Plasmodium berghei NYU-2) infection was studied. The dietary administration of 167 ppm (167 microgram/g) of PCB 1242 or HCB for 3 weeks resulted in an enhanced sensitivity to gram-negative endotoxin (Salmonella typhosa), which was further increased in animals maintained on the diets for 6 weeks. By 6 weeks, a 5.2- or 32-fold increase in endotoxin sensitivity was seen in mice fed PCB or HCB, respectively. A 20% decrease in mean survival time of mice fed PCB 1242 for 3 or 6 weeks and inoculated with malaria was demonstrated. Infected mice that received HCB for 3 or 6 weeks manifested a reduction in mean survival time of 24 or 31%, respectively. Histopathological examination revealed a normal thymus, spleen, mesenteric lymph nodes, and lungs. Centrilobular and pericentral hepatocyte hypertrophy, common to organochlorine exposure, was observed. Electron capture gas chromatographic analysis for PCB 1242 or HCB in the tissues examined histologically revealed a significant deposit of the xenobiotics. HCB concentration was approximately 16 to 25 times greater than that of PCB. The data indicate that environmental chemicals impair host resistance and that the alteration may be related to the presence of the chemicals in the lymphoreticular organs.


Listeria monocytogenes cell walls induce decreased resistance to infection. A significant decrease in murine resistance to Listeria monocytogenes was induced by using crude Listeria cell wall fraction (LCWF) and purified Listeria cell walls (PF). When equal amounts of these materials were injected, PF was more effective than LCWF in decreasing resistance. The PF effect was dose dependent when measured either as a decrease in 50% lethal dose of the Listeria challenge or as a decrease in survival time of the infected mice. PF apparently does not act directly on the Listeria since it (i) did not cause a change in in vitro growth of Listeria and (ii) did not increase the virulence of Listeria passaged in vivo or in vitro. The greatest decrease in resistance was observed when both PF and the Listeria challenge were injected intraperitoneally, which may suggest a localized effect. A decrease in resistance was seen when PF was given as early as 3 days before challenge. There was little or no decrease in resistance when PF was given 2 days after the Listeria challenge. Mice previously immunized with live Listeria were immune to Listeria challenge. However, after PF injection the immune mice showed a decreased resistance which was of the same order of magnitude as that seen in unimmunized mice. The effect of PF seems to be at least partially nonspecific, since a decrease in resistance to Salmonella typhimurium could also be demonstrated in PF-treated mice. Phagocytosis of Listeria both in vivo and in vitro did not appear to be inhibited by PF, although the ability of PF-treated mice to kill Listeria in the peritoneal cavity was inhibited.


Effect of growth environment on Pseudomonas aeruginosa killing by rabbit polymorphonuclear leudocytes and cationic proteins. Pseudomonas aeruginosa grown in a chemostat under carbon- and magnesium-limited conditions showed varying resistance to killing by rabbit peritoneal exudate polymorphonuclear leukocytes. Slow-growing (D = 0.05 h-1), magnesium-limited cells were significantly more resistant to the lethal effects of the phagocytes than were fast-growing magnesium-limited cells and carbon-limited cells (D = 0.05 h-1 and D = 0.5 h-1, respectively). The resistance of magnesium-limited cells to killing by cationic proteins isolated from the leukocytes was shown to be growth-rate dependent, the slowest-growing (D = 0.05 h-1) cells being the most resistant. Carbon-limited cells were sensitive to killing by the cationic proteins at all growth rates tested. Antisera raised in rabbits to all types of cells and commercial anti-Pseudomonas serum rapidly agglutinated magnesium-limited cells but failed to agglutinate carbon-limited cells. There was some indication that slow-growing (D = 0.05 h-1), magnesium-limited cells agglutinated most readily with both types of antisera. No difference was detected in the mouse toxicity of heat-killed cells grown under the various conditions.


Suppression of cellular responses in mice during Trypanosoma cruzi infections. Immunosuppression has been reported to occur in several protozoan parasitic infections. The significance of this suppression on host resistance or on parasite avoidance of immune destruction has not, however, been determined. In the present study two strains of mice that differ with respect to resistance to Trypanosoma cruzi were examined during the course of infection for differences in expression of suppression of blastogenic responses to phytohemagglutinin and an antigen preparation of these parasites. It was found that in vitro blastogenic responses were suppressed in both strains of mice: on day 12 for C57BL/6 mice (resistant strain) and on day 17 for C3H(He) mice (susceptible strain). Neither C3H(He) nor C57BL/6 lymph node cells (LNC) responded to a crude sonically treated antigen of these parasites, although C57BL/6 LNC were inhibited by this antigen later in infection. There was no abrogation of suppression of LNC responses late in infection, when decreases in spleen weight or total spleen cell numbers occurred, in the resistant C57BL/6 mice. LNC from normal uninfected mice were found to be completely suppressed in responsiveness to phytohemagglutinin when cultured at a 2:1 ratio with LNC from C57BL/6 mice with 18-day infections. Attempts to characterize the cell type responsible for this suppression showed it to be a non-thy 1.2-bearing, nylon wool-adherent cell.


Cigarette smoke and phagocyte function: effect of chronic exposure in vivo and acute exposure in vitro. Phagocytic function was studied in mice chronically exposed to cigarette smoke, and the effects of in vitro exposure to cigarette smoke on macrophage activity were also assessed. Cultures of radiolabeled Pseudomonas aeruginosa were employed to investigate phagocyte activity in vivo and in vitro. Mice were exposed on weekdays to fresh cigarette smoke for periods up to 37 weeks and the bactericidal and clearance activity of their lungs was measured. Both pulmonary clearance and bactericidal activity was impaired. The clearance of intravenously injected bacteria from the blood of smoke-exposed mice occurred at the same rate as in control mice, but the accumulation of radiolabel by the liver was decreased. In addition, the rate of elimination of radiolabel from the liver was less than the controls. Macrophages exposed to cigarette smoke in vitro initially had a depressed phagocytic rate, but if phagocytosis over a prolonged period was measured it was eventually enhanced over the rate of control macrophages. The vapor phase of cigarette smoke could also transiently inhibit and then enhance the phagocytic activity.


Induction of capping in blood-stage trypomastigotes of Trypanosoma cruzi by human anti-Trypanosoma cruzi antibodies. The effect of human-specific antibody on surface membrane antigens of trypomastigotes of Trypanosoma cruzi was studied in vitro by using immunofluorescence methods. Immune sera induced aggregation of surface antigens in trypomastigotes to form polar cell caps.


Role of iron in disseminated gonococcal infections. Strains of Neisseria gonorrhoeae isolated from disseminated gonococcal infections have enhanced ability to acquire iron in an experimental host when compared with urogenital gonococcal isolates from patients with uncomplicated infections.


Relationship of method of administration to respiratory virulence of Klebsiella pneumoniae for mice and squirrel monkeys. Klebsiella pneumoniae given by aerosol was significantly less virulent in mice and monkeys than when given by intranasal (mice) or intratracheal (monkeys) instillation.


Detection of the Rocky Mountain spotted fever agent, Rickettsia rickettsii, in dead ticks, Dermacentor andersoni. Rickettsia rickettsii remained viable and retained chromophilic properties for not more than 24 h after Dermacentor andersoni were killed by freezing. Antigenic reactivity was detected for at least 71 days by direct immunofluorescence. However, rickettsiae in ticks suffocated with mineral oil remained pathogenic for at least 14 days. Accordingly, ticks removed from a host by mineral oil or dying from desiccation in transit are still suitable for rickettsial examination.


Adjuvant activity of synthetic 6-O-"mycoloyl"-N-acetylmuramyl-L-alanyl-D-isoglutamine and related compounds. Adjuvant and antitumor activities of synthetic 6-O-"mycoloyl"-N-acetylmuramyl-L-alanyl-D-isoglutamine were examined. All the synthetic 6-O-corynomycoloyl-, 6-O-mocardomycoloyl-, and 6-O-mycoloyl-N-acetylmuramyl-L-alanyl-D-isoglutamine were active as adjuvants for cell-mediated immune responses. However, 6-O-mycoloyl-N-acetylmuramyl-L-alanyl-D-isoglutamine was less active as an adjuvant on circulating antibody formation. It was shown that pyrogenic activity of N-acetylmuramyldipeptide was reduced by 6-O-acylation with mycolic acid, but not with nocardomycolic or corynomycolic acid. Tumor-suppression activity was observed by the synthetic 6-O-mycoloyl-N-acetylmuramyl-L-alanyl-D-isoglutamine by using transplantable tumor in syngenic mice.


Quantitative measurement of phagocytosis of Neisseria gonorrhoeae by mouse peritoneal macrophages. The simultaneous labeling of gonococci with [6-3H]uracil and of mouse peritoneal macrophages with L-[U-14C]leucine permits a quantitative assessment of the association of gonococci with macrophages under various experimental conditions. Colony-type T1 (piliated) gonococci associated more than T4 (nonpiliated) organisms at 4 degrees C, but at 37 degrees C the association of T4 gonococci with macrophages exceeded that for the T1 organisms. The association of T1 gonococci with macrophages could be enhanced as much as 70-fold by homologous rabbit antisera prepared against whole, formaldehyde-treated organisms. This immune enhancement represented primarily increased phagocytosis rather than surface attachment, as shown by its inhibition at 4 degrees C or with 2-deoxyglucose. The data further suggested that this enhanced phagocytosis was mediated via the Fc receptor for immunoglobulin G.


Lysozyme: primary bactericidin in human plasma serum active against Bacillus subtilis. The in vitro bactericidal reaction of human plasma serum against Bacillus subtilis was investigated. Human lysozyme was purified to homogeneity, and antiserum was prepared against the enzyme. The anti-lysozyme immunoglobulin G was used as a specific inhibitor in bactericidal and bacteriolytic reactions. It was found that at low serum concentrations lysozyme was the primary bactericide active against B. subtilis. At appreciably higher serum concentrations, a lysozyme-independent bactericidal activity was also demonstrated.


Demonstration of K88ac and K88ab antigens of Escherichia coli by means of immunoelectrophoresis and immunodiffusion. Five strains of Escherichia coli were tested for the presence of the K88ac or K88ab antigens by immunoelectrophoresis and immunodiffusion. The K88ac antigen of 0A2 and Sojka Abbotstown gave an anodic line in the immunoelectrophoresis test and a line in immunodiffusion with homologous K88ac antisera. The K88ab antigens of 0G7, 0E68, and Moon 263 also gave anodic lines in immunoelectrophoresis, and were detectable by immunodiffusions. The 0 groups of these strains were also demonstrated by immunoelectrophoresis and immunodiffusion with homologous 0 antisera. Lack of complete inactivation at 100 degrees C of both the K88ac and K88ab antigens was noted in this study.


Immunoglobulin biosynthesis in vitro after stimulation of pig blood lymphocytes with mitogen. The biosynthesis of immunoglobulin (Ig) by pig blood lymphocytes after mitogen stimulation with lipopolysaccharide (LPS) and anti-Ig serum has been assessed by incorporation of 3H--leucine into secreted Ig. Lymphocytes prepared from defibrinated and heparinised blood responded to LPS with mitosis, but Ig biosynthesis (predominantly IgM) only occurred with heparinised blood lymphocytes. Anti-Ig treatment produced pronounced mitosis in both preparations but no significant Ig biosynthesis in either. Mitosis was induced by anti-IgM (mu- and L chains), anti-mu and anti-IgG (gamma- and L chains) but only weakly by anti-gamma in both types of preparation. Lymphocytes from heparinised blood gave preparations with higher percentages of surface-Ig-bearing cells by direct immunofluorescence (defibrinated 14.0%, heparinised 25.2%).


Argon laser trabeculotomies in primates: evaluation by histological and perfusion studies. The continuous argon laser beam has been applied to the trabeculum at the anterior chamber angle of one eye of nine primates. The treatment was evaluated by light and scanning electron microscopy and constant rate infusion studies at different intervals following the laser treatment. This histological studies revealed coagulative necrosis, with trabeculotomies extending into Schlemm's canal in the immediate postoperative period. However, complete closure of the laser-induced trabeculotomies by healing was observed within 1 to 3 weeks following the treatment. Outflow facility data as measured by the perfusion studies revealed somewhat increased values of the treated eye during the early 2 weeks after laser treatment. On longer follow-up, no significant outflow differences were measured between the treated and untreated eyes.


Discontinuous counter-immunoelectrophoresis in the study of viruses. A method for the use of the discontinuous counter-immunoelectropnoresis (DCIE) technique in the study of viruses of the major taxonomic groups is reported. The method was successfully employed in detection of antigen of viruses in the bunyavirus, bunyavirus-like, flavivirus, orbivirus, and rhabdovirus groups, and/or of antibody to these agents. Although less sensitive than the complement fixation test, the DCIE test was found to be rapid and in some cases more specific in differentiating related viruses or detecting antigenic similarity between strains of the same virus.


Is the solitary dark neuron a manifestation of postmortem trauma to the brain inadequately fixed by perfusion? Dark neurons, classified as solitary because of their sparse occurrence, were discerned in the transitional zones between gray and white matter in various species of laboratory animals fixed by perfusion. These neurons, histologically indistinguishable from dark neurons in immersion fixed material, tended to develop when the saline perfusion was delayed or slow, the amount of the Bouin fixative was excessive, or the autopsy was performed shortly after the perfusion. Under these conditions, the white matter manifested a softer consistency and a paler color than the gray matter. These observations suggest that, as the consequence of regional differences in intensity and speed of fixation, distortion during extraction of the brain may activate a stress force in the transitional zones where incompletely fixed neurons become affected and acquire an abnormal affinity for aniline dyes and silver.


Catalysis of CO2 reactions by lung carbonic anhydrase. Steady-state carbon dioxide excretion was studied in isolated bloodless lung preparations perfused with bicarbonate solutions. Addition of acetazolamide produced a prompt, significant decrease in the volume of excreted CO2 under all conditions studied. Excreted CO2 was derived from two sources: CO2 dissolved in the perfusate and CO2 produced by dehydration of bicarbonate in the pulmonary capillary. The relative quantity of these two sources was determined by measurement of the simultaneous excretion of acetylene. Determination of the rate of CO2 production permitted the calculation of capillary blood volume, mean capillary transit time, and the degree of catalysis of CO2 reactions by carbonic anhydrase present in the lung. Contamination of perfusate with blood carbonic anhydrase was ruled out by measuring hemoglobin concentration and carbonic anhydrase activity in pulmonary venous drainage. Comparison of steady-state CO2 production during control conditions and carbonic anhydrase inhibition indicated that bicarbonate in plasma has access to sufficient lung carbonic anhydrase to catalyze the CO2 hydration-dehydration reaction by a factor of five.


Nonrebreathing valve for spontaneously breathing or mechanically ventilated animals. The design of a small dead space, nonbreathing valve suitable for use in small laboratory animals is presented. The design overcomes previous problems of condensation occurring at valve leaflets, and allows ongoing collection of expired gases should the animal require mechanical ventilation. The valve apparatus performs reliably when in operation for several hours at a time with a variety of small animals.


Motorized minitrack for exercising large animals. A simply constructed device for exercise training large animals, e.g., swine, is described. Basically the device is a small, circular, enclosed track constructed of plywood and metal fence material. A centrally located, vertical shaft powered by a DC motor rotates at a variable speed. Attached to each of four horizontal metal beams extending from the shaft are four suspended partitions which hang freely above the enclosed track surface. As the shaft rotates, the four partitions move in unison, thus forcing animals in each compartment to move at the same pace. The advantages of this device are its cost, low maintenance, and the flexibility to train eight animals by one person. Preliminary results of a study on training pigs are discussed.


The Behavior Observation Instrument: a method of direct observation for program evaluation. The background and development of a multicategory direct observation system, the Behavior Observation Instrument (BOI), is described. This time-sampling procedure for recording the behavior of persons is demonstrated in several treatment settings and the results applied to issues of program evaluation. Elements that have prevented direct observation from being widely adopted, such as costs, manpower, and training requirements, are systematically analyzed. A basic psychometric analysis of the instrument is used to determine optimum frequency and duration of observation intervals as well as observer agreement. The results imply that direct observation methods, once assumed by some to belong to the special province of the single-subject design, can be used to assess the effects of programs on groups of psychiatric clients in an efficient and economic manner.


Post-transcriptional modifications of the anticodon loop region: alterations in isoaccepting species of tRNA's during development in Bacillus subtilis. Structural similarities of tRNA's were compared using three sets of isoaccepting species that had previously been shown to undergo significant changes in chromatographic elution properties as a function of developmental stage in Bacillus subtilis. Comparisons of the structures of the tRNA's were based on the composition of their modified nucleosides, comparisons of oligonucleotide elution profiles from RPC-5 columns, and two-dimensional electrophoretic fingerprint analysis of oligonucleotides. The tRNA's studied were tRNA(Lys) (1) and tRNA(Lys) (3); tRNA(Tyr) (1) and tRNA(Tyr) (2); and tRNA(Trp) (1) and tRNA(Trp) (2). The results suggest that the difference among these pairs of isoaccepting species is a difference in the degree of post-transcriptional modifications of the anticodon loop region. The nucleosides involved were N(6)-(Delta(2)-isopentenyl)adenosine (i(6)A), 2-methylthio-N(6)-(Delta(2)-isopentenyl)adenosine (ms(2)i(6)A), and an unknown nucleoside K, which occurred in a position analogous to N-[9-(beta-d-ribofuranosyl)purin-6-ylcarbamoyl]threonine. The amounts of i(6)A and ms(2)i(6)A, determined using total tRNA from exponential-or stationary-phase cells, suggest that the thiomethylation of i(6)A is a pleiotropic phenomenon affecting several tRNA species. As opposed to the situation in Escherichia coli tRNA, where ms(2)i(6)A constitutes about 90% of the total hydrophobic nucleosides at all growth stages, B. subtilis tRNA's have i(6)A as the predominant hydrophobic nucleoside in exponential growth and ms(2)i(6)A as the predominant nucleoside in stationary phase. Thus, the enzyme system which forms i(6)A and the enzyme system which thiomethylates i(6)A are not coordinated during growth in B. subtilis as they are in E. coli. It is suggested that these changes in anticodon loop modifications in B. subtilis may be related to changes in the translational apparatus which occur during sporulation.


Study of calcium dipicolinate release during bacterial spore germination by using a new, sensitive assay for dipicolinate. The release of calcium and dipicolinic acid from spores of Bacillus megaterium KM during L-alanine-induced triggering of germination has been studied using a new, simple, and rapid assay for dipicolinic acid capable of detecting a concentration of 0.5 micron. The release of both calcium and dipicolinate started within seconds of exposure of the spores to L-alanine, thus preceding other measurable changes associated with germination. From the earliest times, the two substances were released in equimolar quantities, although later in germination calcium predominated.


Cellular lysis of Streptococcus faecalis induced with triton X-100. Lysis of exponential-phase cultures of Streptococcus faecalis ATCC 9790 was induced by exposure to both anionic (sodium dodecyl sulfate) and nonionic (Triton X-100) surfactants. Lysis in response to sodium dodecyl sulfate was effective only over a limited range of concentrations, whereas Triton X-100-induced lysis occurred over a broad range of surfactant concentrations. The data presented indicate that the bacteriolytic response of growing cells to Triton X-100: (i) was related to the ratio of surfactant to cells and not the surfactant concentration per se; (ii) required the expression of the cellular autolytic enzyme system; and (iii) was most likely due to an effect of the surfactant on components of the autolytic system that are associated with the cytoplasmic membrane. The possibility that Triton X-100 may induce cellular lysis by releasing a lipid inhibitor of the cellular autolytic enzyme is discussed.


Two bacteriophages which utilize a new Escherichia coli major outer membrane protein as part of their receptor. Escherichia coli strain JF694 contains a new major outer membrane protein which we have called protein E (J. Foulds, and T. Chai, J. Bacteriol. 133:1478-1483). Two new bacteriophages, TC45 and TC23, were isolated that require the presence of protein E in the outer membrane of host cells for growth. Both of these bacteriophages have a morphology similar to T-even bacteriophages but are distinct in properties such as plaque morphology, buoyant density, and burst size. Although strain JF694, containing protein E, adsorbs bacteriophage TC45 efficiently, cells killed with heat or chloroform are unable to inactivate this bacteriophage. Purified protein E either in the presence or absence of additional probable cofactors such as lipopolysaccharide was also unable to inactivate bacteriophage TC45. Both bacteriophages probably use protein E as at least part of their receptor but require, in addition, other outer membrane components or a specific orientation or organization of this protein in the outer membrane.


F'-plasmid transfer from Escherichia coli to Pseudomonas fluorescens. Various F' plasmids of Escherichia coli K-12 could be transferred into mutants of the soil strain 6.2, classified herein as a Pseudomonas fluorescens biotype IV. This strain was previously found to receive Flac plasmid (N. Datta and R.W. Hedges, J. Gen Microbiol. 70:453-460, 1972). ilv, leu, met, arg, and his auxotrophs were complemented by plasmids carrying isofunctional genes; trp mutants were not complemented or were very poorly complemented. The frequency of transfer was 10(-5). Subsequent transfer into other P. fluorescens recipients was of the same order of magnitude. Some transconjugants were unable to act as donors, and these did not lose the received information if subcultured on nonselective media. Use of F' plasmids helped to discriminate metabolic blocks in P. fluorescens. In particular, metA, metB, and argH mutants were so distinguished. In addition, F131 plasmid carrying the his operon and a supD mutation could partially relieve the auxotrophy of thr, ilv, and metA13 mutants, suggesting functional expression of E. coli tRNA in P. fluorescens. In P. fluorescens metA Rifr mutants carrying the F110 plasmid, which carried the E. coli metA gene and the E. coli rifs allele, sensitivity to rifampin was found to be dominant at least temporarily over resistance. This suggests interaction of E. coli and P. fluorescens subunits of RNA polymerase. his mutations were also complemented by composite P plasmids containing the his-nif region of Klebsiella pneumoniae (plasmids FN68 and RP41). nif expression could be detected by acetylene reduction in some his+ transconjugants. The frequency of transfer of these P plasmids was 5 X 10(-4).


Ultrastructural study of Salmonella typhimurium treated with membrane-active agents: specific reaction dansylchloride with cell envelope components. Amino groups of cell envelope proteins, lipids, and lipopolysaccharides cannot be labeled in intact cells of Salmonella typhimurium G 30 by using 5-dimethylaminonaphthalene-1-sulfonylchloride incorporated in lecithin-cholesterol vesicles. However, application of membrane-interacting agents like tris(hydroxymethyl)aminomethane (Tris)-hydrochloride, ethylenediaminetetraacetate (Na salt) (EDTA), divalent cations, and sublethal doses of the cationic antibacterial agents polymyxin B and chlorhexidine induced specific fluorescent labeling of envelope proteins and lipids but not of cytoplasmic compounds, with the exception of a soluble protein with a molecular weight of 46,000 in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Treatment with Tris-hydrochloride buffer produced labeling of the heat-modifiable protein B/B(+) and of proteins with molecular weights of 26,000, 22,000, and below 17,000. A combination of Tris-hydrochloride and EDTA induced additional dansylation of the major protein A and of proteins of molecular weights 80,000, 60,000, and 44,000. Polymyxin B and chlorhexidine caused similar labeling patterns. In every case, except with divalent cation treatment, protein B/B(+) was the most prominently labeled species. Phosphatidylethanolamine was dansylated up to 30%. Lipopolysaccharide was not reactive under any condition or treatment. In addition, the peptidoglycan-bound lipoprotein did not react with dansylchloride in either intact or Tris-hydrochloride-treated cells. The results are discussed with regard to a possible localization of labeled and unlabeled compounds of the cell envelope on the basis of a model placing cell envelope amino groups into ion-ion interactions with anionic components of other envelope compounds like phosphate and carboxyl groups.


Isolation of large bacterial plasmids and characterization of the P2 incompatibility group plasmids pMG1 and pMG5. Large plasmids from Agrobacterium tumefaciens, Salmonella typhimurium, Escherichia coli, Pseudomonas putida, and Pseudomonas aeruginosa were routinely and consistently isolated using a procedure which does not require ultracentrifugation but includes steps designed to separate large-plasmid DNA from the bacterial folded chromosome. It also selectively removes fragments of broken chromosome. A variety of large plasmids was readily visualized with agarose gel electorphoresis, including five between 70 and 85 megadaltons (Mdal) in size, six between 90 and 143 Mdal, one that was larger than 200 Mdal, and one that was larger than 300 Mdal. This isolation procedure allowed initial estimation of the molecular sizes of the two IncP2 plasmids, pMG1 and pMG5, which were 312 and 280 Mdal, respectively. A standard curve for size determination by gel electrophoresis including plasmids between 23 and 143 Mdal in size did not extrapolate linearly for plasmids of the 300-Mdal size range. Unique response of different plasmids to the isolation procedure included sensitivity of IncP1 plasmids to high pH and the co-isolation of a 20-Mdal "cryptic" plasmid in conjunction.


Identification of a new sequence-specific endonuclease, NgoII, from Neisseria gonorrhoeae. A class II restriction endonuclease which recognizes the same nucleotide sequence as EndoR.HaeIII has been found in four of seven isolates of Neisseria gonorrhoeae tested.


TOL is a broad-host-range plasmid.

TOL is a broad-host-range plasmid. We readily isolated insertions of the carbenicillin resistance element Tn401 into the TOL plasmid in Pseudomonas putida. Hybrid TOL::Tn401 plasmids stably express the Cbr phenotype in Pseudomonas aeruginosa and Escherichia coli. Whereas the replicative and conjugative functions are expressed in both hosts, the ability to grow on m-toluate is only expressed in the Pseudomonas species.


Immunochemical comparison between an oxacillin-hydrolyzing penicillinase of Aeromonas hydrophila and those mediated by R plasmids. Antiserum against an oxacillin-hydrolyzing penicillinase of Aeromonas hydrophila did not show immunochemical cross-reaction with four oxacillin-hydrolyzing penicillinases mediated by R plasmids.


Prevention of autolysis in suspensions of Neisseria gonorrhoeae by mercuric ions. Mercuric ions were the only metal ions which prevented autolysis of Neisseria gonorrhoeae, as measured both by stabilization of optical density and by prevention of release of [3H]diaminopimelic acid.


Biosynthesis of delta-aminolevulinic acid by blue-green algae (cyanobacteria). When levulinic acid, a competitive inhibitor of delta-aminolevulinic acid dehydratase, was added to growing cultures of blue-green algae (cyanobacteria), delta-aminolevulinic acid was excreted into the medium and cell growth was inhibited.


Physiological role of oxygenated cytochrome o: observations on whole-cell suspensions of Vitreoscilla. The form of cytochrome o that predominates in Vitreoscilla cells having various levels of respiratory activity was studied by using untreated, frozen-thawed, and starved cells, which had respiratory rates decreasing in the order given. Direct spectral observation revealed that the oxygenated form of cytochrome o predominated during the aerobic steady-state oxidation of endogenous substrate or exogenous glutamate in untreated and frozen-thawed cells and was replaced by the reduced form when the cell suspensions became anaerobic. The respiratory rates, estimated inversely from the time of duration of the steady state, were correlated to the rates of oxygen consumption for the various cells. Oxidized cytochrome o predominated in aerobic starved cells. These results indicate the involvement of three forms of cytochrome o--oxidized, reduced, and oxygenated--in the catalytic and cyclic change of this cytochrome. The oxygenated form also appeared after the addition of hydrogen peroxide to the cells, but only the oxidized form appeared when ethyl hydrogen peroxide was added. The appearance of the oxygenated form with the addition of hydrogen peroxide was probably due to the reaction of the reduced cytochrome with the oxygen that had evolved by the action of catalase present in the cells.


Polyethylene glycol-induced fusion of heat-inactivated and living protoplasts of Bacillus megaterium. Protoplasts of Bacillus megaterium, incubated at 50 degrees C for 120 min, lost the ability to revert to bacillary form. Such heat-inactivated protoplasts, however, produced recombinants when fused by polyethylene glycol treatment with normal protoplasts. Although this differential inactivation effect is not yet fully reproducible, reciprocal inactivations of the parental protoplasts in genetic crosses have clearly shown that for protoplast fusion (i) either of the parents may serve as the viable recipient for markers coming from the heated parental protoplasts, and (ii) either of the parents may be rendered nonviable and yet, when fused with a viable partner, contribute to formation of a recombinant. Heat inactivation seems to provide a way to counterselect when few markers are available and one of the parents is prototrophic.


Repression of biotin biosynthesis in Escherichia coli during growth on biotin vitamers. A strain of Escherichia coli in which the lacZ gene was fused to the bioA promoter was constructed. Colonies of this strain formed Lac(+) colonies on low-biotin agar (1.6 to 4.1 nM) and Lac(-) colonies on high-biotin agar (41 nM). This lac-bio fusion strain was used to study the question of whether cells growing on the biotin vitamers d-biotin-d-sulfoxide (BDS) and dethiobiotin (DTB) generate enough biotin to give maximal repression of beta-galactosidase synthesis. Repression by high concentrations (400 nM) of BDS was almost maximal (about 96%), whereas DTB repression reached a saturation level of about 80% with increasing DTB concentrations. The levels of repression obtained with both vitamers were sufficient to cause the colonies to appear Lac(-). When the lac-bio fusion was transduced into lines carrying mutations (bis) that prevent reduction of BDS to biotin, the transductants were not repressed by added BDS. Repression by BDS is unlikely to result from accumulation of extracellular biotin-related substances because (i) washed bis(+) cells were not detectably derepressed when transferred into medium containing BDS and (ii) washed bis cells were not detectably repressed when transferred into medium in which bis(+) cells had grown. Lactose agar plates containing high concentrations of DTB or BDS comprise an efficient selective medium for bioB or bis mutants and were used to isolate spontaneous mutations of these genes. This method should be adaptable to the selection of mutations in any biosynthetic pathway subject to end-product repression.