Side-to-side portacaval shunt in the treatment of Budd-Chiari syndrome. This is a case of a 22-year-old female patient with idiopathic thrombosis of the hepatic veins in whom side-to-side portacaval shunt was followed by the disappearance of a hitherto intractable ascites and a marked improvement in her general condition. The patient remained well for 1 1/2 years after surgery; however, liver cirrhosis, which was present before shunt, persisted. From this case and from the few published cases, the authors concluded that side-to-side portacaval shunt might be beneficial to patients with thrombosis of the hepatic veins.


Vasodilatory drugs in the management of nonocclusive bowel ischemia. A patient with a clinical and angiographic diagnosis of nonocclusive mesenteric ischemia had phenoxybenzamine directly infused into the superior mesenteric artery to counteract the existing splanchnic vasoconstriction. The diagnosis was confirmed by later operative intervention and the patient recovered. Vasodilatory drugs may play a significant role in the management of patients with nonocclusive mesenteric ischemia.


Salmonella heidelberg infection in Caroli's syndrome. A PATIENT IS DESCRIBED IN WHOM DIARRHEA AND FEVER DUE TO Salmonella heidelberg was followed by a series of episodes of cholangitis due to the same organism. Cholecystectomy and prolonged courses of antibiotics failed to eradicate the infection, and the presence of multiple intrahepatic biliary cysts (Caroli's syndrome) was eventually discovered by endoscopic retrograde cholangiography.


An electron-microscopic study of jejunal mucosal morphology in control subjects and in patients with tropical sprue in southern India. The ultrastructure of the jejunal mucosa obtained by peroral biopsy from 7 control subjects and 12 patients with tropical sprue was studied. Compared with biopsies from control subjects in temperature zones, southern Indian control subjects showed minimal increase in lysosomes in surface epithelial cells, an occasional dark-staining degenerating epithelial cell in the upper two-thirds of villi, and an increase in cellular infiltration of surface epithelium and lamina propria. The jejunal epithelial cells of tropical sprue patients had abnormal microvilli, marked increase in lysosomes, increase in intracellular fat, and degenerative changes in rough endoplasmic reticulum and mitochondria. Two types of degenerating epithelial cells, designated pale and dark based on their staining quality, were seen in the villi. Pale staining degenerating cells were also seen in the crypts. The basement membrane region showed an amorphous deposit with many collagen fibers and fat droplets. The epithelial layer and lamina propria showed increased cellular infiltration. In one biopsy from a patient with sprue, particles of possible viral origin were seen.


Sequelae of colectomy and ileostomy: comparison between Crohn's colitis and ulcerative colitis. A comparison is made of the immediate and long term mortality of colectomy and ileostomy between 73 patients who had Crohn's colitis and 442 who had ulcerative colitis. The immediate mortality in Crohn's disease is 4%. In ulcerative colitis it is 10%, chiefly because of the higher proportion of emergency operations. The late mortality in both groups is 10%, chiefly as a result of recurrence of Crohn's disease or the sequellae of colonic malignancy present at the time of colectomy for ulcerative colitis. A further comparison is made between the postoperative course of the 64 surviving patients with Crohn's disease and a comparable sample of 65 patients who had an ileostomy for ulcerative colitis in the same era. There was a similar incidence of postoperative septic complications in the two groups (35%). The readmission rate was twice as high in the Crohn's disease patients. Ileostomy reconstruction for mechanical complication was needed in 21 patients with Crohn's disease compared with 6 with ulcerative colitis. Further ileal resection was required for recurrent disease on another 25 occasions in the patients with Crohn's disease but never in those with ulcerative colitis. Long therm review graded the clinical status as excellent or good in 70% of those with Crohn's disease compared with 95% with ulcerative colitis.


The normal human esophageal mucosa: a histological reappraisal. In 19 asymptomatic subjects, a total of 95 mucosal suction biopsies were taken from multiple sites in the distal 10 cm of esophagus. The biopsies were examined for evidence of basal cell hyperplasia and elongated dermal papillae, features considered to be histological consequences of gastroesophageal reflux. Fifty-seven per cent of the biopsies in the distal 2.5 cm of the esophagus and 19% of the biopsies above 2.5 cm exhibited these histological features.


Effect of sodium, mannitol, and magnesium on glucose, galactose, 3-O-methylglucose, and fructose absorption in the human ileum. Although it is generally agreed that active sugar absorption in vitro is absolutely dependent on the presence of sodium ions on the luminal side of the mucosa, previous in vivo studies in the ileum of rat, dog and man have shown that active glucose absorption is almost as rapid from a sodium-free mannitol solution as from a sodium chloride solution. These experiments were performed in hopes of reconciling this discreptancy. Absorption of three actively transported sugars (glucose, galactose, and 3-O-methylglucose) having different apparent Km's, and of fructose (absorbed by a separate carrier-mediated process) were measured in the human ileum in vivo. The following observations were made: (1) Mannitol substitution for sodium results in only a slight reduction (23%) in the active absorption of glucose. (2) Magnesium substitution for sodium results in a greater depression (45%) of glucose absorption. (3) The apparent Km for glucose absorption is increased when sodium is replaced by magnesium, but the Vmax is not altered. (4) Magnesium does not depress glucose absorption or the apparent Km for glucose transport when sodium is present in the perfusing solution. (5) Neither sodium removal nor magnesium has any effect on fructose absorption. (6) Absorption of galactose and 3-O-methylglucose (low affinity sugars for the glucose carrier) is reduced by about 40 to 50% when mannitol replaces sodium, but magnesium substitution for mannitol in a sodium-free medium does not further depress absorption of these sugars. The following conclusions are suggested by these results: First, part of the discrepancy between previous in vitro and in vivo experiments was due to the type of test sugar (glucose versus glucose analogue) and the solute used to replace sodium in the luminal solution. Second, magnesium is more effective than mannitol in reducing sodium concentration at the glucose transport site on the brush border. Third, luminal sodium ions have an important effect on active sugar absorption in the human small intestine in vivo, as they do in vitro. And, fourth, there is a component of active sugar absorption (about one-half) which appears to be independent of luminal sodium ions in vivo.


Increased sensitivity of the cholylglycine breath test for detecting ileal dysfunction. Deconjugation of bile salts was assessed by measuring the rate of appearance of 14-CO2 in the breath after the administration of cholyl [1-14-C]glycine to patients with ileal dysfunction (ileitis or ileal resection), cholesterol cholelithiasis with or without cholecystectomy, and control subjects (healthy or with diarrhea). Vitamin B12 absorption was also measured in many of the patients with ileal dysfunction by determining the recovery of 57-Co in the urine (Schilling test). The percentage of administered 14-C excreted in breath 14-CO2 was increased in the patients with ileal dysfunction, but there was considerable overlap between the patients with ileal dysfunction and the control subjects. Breath 14-CO2 was excreted more rapidly in all the patients with ileal dysfunction than in the control subjects. Of the calculated 24-hr breath excretion 78.3 plus or minus 8.9% SD of administered 14-C was excreted in 12 hr in the patients with ileal dysfunction, compared to 39.1 plus or minus 13.1% in the control subjects; only 1 of the 19 patients with ileal dysfunction overlapped with the control subjects. The Schilling test was normal orequivocal in 5 of 14 patients with ileal dysfunction. The percentage of administered 14-C excreted in breath 14-CO2 was increased in patients with cholecystectomy, but excretion was not more rapid than in the control subjects. The data suggest that the sensitivity of the breath test for diagnosing ileal dysfunction may be usefully increased by determining the rate of excretion of breath 14CO2.


Atrophy of villi with hypertrophy and hyperplasia of Paneth cells in isolated (thiry-Vella) ileal loops in rabbits. Light-microscopic studies. Thiry-Vella loops in rabbit ileum were prepared by a new technique and were studied 18 hr to 49 days postisolation. The loops became grossly shortened after 14 days. Histologically, some shortening and blunting of villi was detectable as early as 4 days postisolation, and with prolonged isolation the changes became marked. Reduction in epithelial cell height and in brush border thickness were noted, and goblet cells were increased somewhat in size and prominence. Yet there was only slightly increased chronic inflammation in the mucosa and acute inflammation was uncommon, suggesting that mucosal injury was minimal. Furthermore, mean epithelial mitotic indices for the crypts did not rise and were generally reduced. Striking hyperplasia and hypertrophy of Paneth cells associated with mitotic figures in Paneth cells accompanied the atrophic changes in the villi. Reimplantation of loops into the bowel 3 weeks after isolation led to complete reversal of all changes, including hyperplasia of Paneth cells. On the other hand, regular perfusion of loops with a solution containing a large variety of nutrient substances failed to reverse the mucosal changes. It was concluded that atrophy of villi in isolated ileum of the rabbit occurred mainly because one or more substances contained in the chyme are needed to maintain normal mucosal architecture. These substances probably help regulate epithelial cell turnover and may well be endogenous in origin. Loss of substances in the chyme after loop isolation may also have led to Paneth cell hyperplasia. Alternatively, the Paneth cell changes and atrophy of villi might have been related in a cause and effect way.


Adenylate cyclase in intestinal crypt and villus cells: stimulation by cholera enterotoxin and prostaglandin E1. The secretory responses to cholera enterotoxin and prostaglandin E1 (PGE1) are dependent upon elevation of the intracellular levels of cyclic AMP. Although several previous reports have suggested that intestinal secretion due to cholera enterotoxin and PGE1 may be confined to the crypt cells, this matter has been incompletely resolved. These studies were undertaken to define the activity of adenylate cyclase in villus and crypt cells from rabbit and rat intestine, and to determine the influence of enterotoxin and PGE1 on this activity, Mucosal fractions were prepared from rabbit ileum with a planing device, and from rat distal small intestine by a vibration technique. In both species base line adenylate cyclase activity was greater in crypt than in villus cells. After exposure to cholera enterotoxin in vivo, adenylate cyclase activity was enhanced in all fractions prepared from rabbit ileum, and the response was most marked in villus cells. Furthermore, adenylate cyclase in membranes prepared from both rat villus and crypt intestinal cells was responsive to the in vitro addition of PGE1. The results of these studies indicate that both villus and crypt cells contain one of the important components required for the cyclic AMP-mediated secretory response, namely, a cholera enterotoxin and PGE1-sensitive adenylate cyclase activity. Since an increased level of cyclic AMP alone may not be sufficient to evoke a secretory response, these studies do not clarify the extent to which each of these major cell types may participate in cyclic AMP-mediated secretion.


Amino terminal gastrin fragment in serum of Zollinger-Ellison syndrome patients. In addition to the previously described molecular forms of gastrin, a new component has been found in high concentrations in the serum of 6 patients with Zollinger-Ellison syndrome. Serum was fractionated by gel filtration, and ion exchange chromatography, and the new component was identified in eluates by radioimmunoassay using an antibody with specificity for the N-terminal portion of heptadecapeptide gastrin. The precise chemical nature of the new component is not known, but its chromatographic behavior and its reactivity to various antibodies is indistinguishable from that of the natural or synthetic N-terminal 1 to 13 fragment of G-17. The new component is present in gastrinoma tumor tissue. Its concentration in serum of gastrinoma patients increases markedly when secretin is injected.


Phlegmonous gastritis.

Phlegmonous gastritis. We have reviewed 23 documented cases of phlegmonous gastritis reported since 1945 in the American literature, and have added 2 of our own. A small series of cases makes it somewhat difficult to draw any definite conclusions regarding the modes of presentation; nonetheless, some clinical trends are discernible. In a patient with a history of large ethanol intake, a recent bout of "gastritis," or recent upper respiratory infection, who presents with acute upper abdominal pain, peritonitis, purulent ascitic fluid, and fever, the diagnosis of phlegmonous gastritis must be considered in differential diagnosis. With normal serum amylase, no historical evidence of ulcer or gallbladder disease, the diagnosis becomes even more probable. Preoperative diagnosis is rare, but gastroscopy with or without biopsy, and culture of gastric contents may establish the diagnosis. The definitive treatment would appear to be resection or drainage of the stomach, combined with large doses of systemic antibiotics, usually penicillin. The surgical mortality in cases reviewed was 18.2%, while the medical mortality was 100%. The overall mortality was 67%. It is hoped that more frequent recognition of this disease entity will lead to earlier diagnosis and a resulting lower morbidity and mortality.


Malabsorption of essential amino acids in tropical sprue. The absorption of seven amino acids from an equimolar mixture containing each of the eight essential amino acids in a concentration of 20 mM was evaluated using a marker perfusion technique in 9 patients with untreated tropical sprue and in 6 healthy Puerto Rican control subjects. The rate of absorption of from two to six of the amino acids tested was lower in 7 of the patients than in the control subjects. The absorption of isoleucine, leucine, valine, and threonine was significantly lower in the group of sprue patients than in the control subjects. Treatment of 4 patients with oral tetracycline for 2 weeks was associated with return of amino acid absorption to normal in the 3 persons who had improved intestinal function, as tested by other parameters. More prolonged treatment was required for normalization of amino acid absorption in the 4th patient. These observations indicate that the absorption of certain essential amino acids is reduced in some patients with untreated tropical sprue.


Coagulation studies in ulcerative colitis and Crohn's disease. Thromboembolic disease is a known complication of ulcerative colitis, but is rarely reported in Crohn's disease. In this study, we have tried to determine if there are indeed coagulation abnormalities associated with these two diseases. Four groups of patients were studies: (1) patients with ulcerative colitis; (2) patients with Crohn's disease; (3) patients with other diseases; (4) healthy control subjects. In both disease groups, platelet counts, factor V, factor VIII, and fibrinogen levels were found to be elevated, while antithrombin III levels were decreased.


Jejunal perfusion of simple and conjugated folates in man. The technique of human jejunal perfusion was used to study the process of digestion and absorption of conjugated folates in five healthy volunteers. The test solution of isotonic saline contained equimolar concentrations of purified [3H]pteroylmonoglutamate ([3H]PG-1) and of [14C]pteroylheptaglutamate ([14C]PG-7) which was labeled on the first glutamyl unit. Calculations were made of the luminal disappearance of each labeled folate, and degradation products of [14C]PG-7 were identified in luminal contents obtained 15 and 30 cm from the infusion site. The percentage of disappearance of [3H]PG-1 was 74.7% and of [14C]PG-7 was 52.6% (P less than 0.001)9 Column chromatography of intestinal aspirates demonstrated a spectrum of 14C-labeled folates corresponding to chain lengths from [14C]PG-1 to [14C]PG-7, with distal accumulation of derived [14C]PG-1. Intraluminal hydrolysis of [14C]PG-7 was excluded by finding the compound unchanged after in vitro incubation with intestinal juice obtained by saline perfusion or siphonage. These data indicate that there are separate rates for the luminal disappearance of PG-1 and PG-7, and show that the digestion of PG-7 requires its contact with the intestinal mucosa. The evidence suggests that progressive mucosal hydrolysis is integral to the process of PG-7 absorption in man.


Changes in liver and gastric mucosal hexosamine synthesis after restraint. The specific activity of L-glutamine: D-fructose-6-phosphate aminotransferase was measured in the oxyntic gland mucosa and liver of fasted, male rats after immobilization in a cold environment. Immobilization for 3 hr resulted in 100% frequency of lesion formation associated with decreased enzyme activity in oxyntic gland mucosa (70.1 plus or minus 5.9% of control) and liver 25.2 plus or minus 5.2% of control). Enzyme activity had returned to control level in the stomach 9 hr after immobilization, and in the liver 21 hr after immobilization. Immobilization for 1 1/2 hr decreased oxyntic gland mucosa enzyme activity to 21.0 plus or minus 9.8% of control, although the frequency of lesion formation was only 62.5%. Liver enzyme activity was 51.3 plus or minus 12.8% of control. Concentrations of UDP-N-acetylhexosamines, feedback inhibitors of this enzyme, were not altered in either tissue. Adrenalectomy, which increased the frequency of damage after 1 hr of immobilization, enhanced the decreases in enzyme activity in both tissues, while atropine sulfate, which decreased the frequency of damage after 3 hr of immobilization, had no significant effect on the enzyme. Thus, immobilization of rats in a cold environment decreases or alters the synthesis of hexosamine-containing compounds through a mechanism not dependent on acid secretion or adrenocorticoids. This alteration in synthesis may result not only in abnormal mucus secretion, but also in altered cell membrane structure and function.


Response of the isolated perfused stomach of the dog to electrical vagal stimulation. The left vagal trunk of blood-perfused, isolated, canine stomach was continously stimulated electrically for periods of 30 min at 7 v and 5-msec duration; the frequency of stimulation was varied without interruption from 0.5 to 10 Hz. Plasma immunoreactive gastrin (IRG), total gastric blood flow (GBF), and gastric acid output (GAO) were monitored continuously. When the vagus was stimulated, gastrin concentrations and GBF increased rapidly, while GAO rose at a slower rate. Two Hertz appeared to be the optimal stimulus frequency for increasing plasma IGR levels, while 10 Hz was most effective for increasing GBF. Increasing the frequency from 0 to 0.5 Hz produced the maximum observed change in GAO; additional increments to 2 or 10 Hz produced no consistent additional increase in GAO. The results indicate that plasma IRG concentration and GBF increase in response to various frequencies of vagal stimulation. Vagal stimulation increases the output of immunoreactive gastrin, as well as acid, from the isolated canine stomach.


Vagal control of gastric electrical control activity and motility. The nerves of Latarjet and those going directly to the fundus and the body of the stomach were stimulated to determine their effect on gastric electrical control activity (ECA) and contractions. The lower limits of effective stimulation parameters were: pulse frequency 2 pulses per sec, pulse width 2 msec, pulse amplitude 0.3 ma or 5 v. Stimulation at or near the lower limits of stimulation parameters caused contractions with little effect on gastric ECA. At higher pulse repetition rates and amplitudes, vagal stimulation caused premature control potentials (PCP's), delayed control potentials (dcp's), and contractions throughout the electrically active region. The PCP's and DCP's in the antrum became phase-unlocked from those in the corpus. After 2 to 5 min of such vagal stimulation, PCP's and DCP's disappeared; gastric control waves became phase-locked at normal frequency and aboarad phase lag. Contractions continued to occur. Atropine and hexamethonium blocked all effects of vagal stimulation. After gastric ECA became insensitive to vagal stimulation, PCP's could still be produced by intraarterial acetylcholine or dimethylphenylpiperazinium, or by an electrical stimulus applied to gastric muscle, and physostigmine could temporarily restore ECA sensitivity to vagal stimulation. The ECA-insensitive state to vagal stimulation represents depletion of acetycholine at preganglionic sites.


Effects of low dose chenodeoxycholic acid feeding on biliary lipid metabolism. To better define the mechanisms by which chenodeoxycholic acid (CDCA) alters the lipid composition of bile and promotes dissolution of cholesterol gallstones, we have studied the effects of relatively low doses of CDCA on the metabolism of biliary lipids in 10 subjects without gallstones. In 5 subjects, the effects of CDCA feeding were compared to those of cholic acid, a bile acid that apparently does not dissolve cholesterol stones. The following measurements were carried out during control and treatment periods: (1) composition of biliary lipids, (2) hepatic secretion rates of biliary lipids, (3) pool sizes of bile acids, and (4) specific composition of bile acids. Low doses of CDCA consistently reduced the lithogenicity of gallbladder bile by decreasing the proportion of cholesterol relative to the solubilizing lipids--bile acids and lecithin. This decrease in lithogenicity was associated with a selective reduction in hepatic secretion rates of cholesterol. At the doses of CDCA given, secretion rates of bile acids and lecithin and pool sizes of bile acids were not significantly changed; also conversion of cholesterol into bile acids was not completely inhibited. Cholic acid feeding appeared to increase the total size of the bile acid pool, but it did not affect the lithogenicity of bile or secretion rates of cholesterol. The data show that at low doses CDCA lowers lithogenicity of bile by reducing hepatic secretion of cholesterol, while cholic acid does not have a similar effect.


Bile acid metabolism in cirrhosis. IV. Characterization of the abnormality in deoxycholic acid metabolism. Several recent studies have demonstrated that patients with cirrhosis frequently lack deoxycholic acid in bile and plasma. In order to explain this observation, comparative experiments on the colonic absorption of deoxycholic acid and on the colonic conversion of cholic to deoxycholic acid were carried out in the cirrhotic patients with normal and very low percentages of deoxycholic acid. Deoxycholic or cholic acid (100 mg) plus 5 muc of each [14C] bile acid were administered by enema to 8 patients with and 5 without liver disease. Deoxycholic acid produced a significant increase in the percentage of biliary deoxycholic acid in patients with cirrhosis. However, the rate of appearance of 14C-deoxycholic acid in patients with cirrhosis was slower than in normal control subjects. Distribution of the 14C activity among the bile acids indicated that rehydroxylation of deoxycholic to cholic acid did not occur. The distribution of 14C activity in biliary bile acids after the rectal administration of [14C]cholic acid showed that patients with severe cirrhosis converted [14C]cholic to [14C]deoxycholic acid at a much slower rate than did cirrhotic patients with normal percentages of biliary deoxycholic acid. Feeding of cholic acid (450 mg per day) for 3 days to 4 cirrhotic patients resulted in a 2-fold increase in the percentage of biliary cholic acid, but only a small increase in the percentage of deoxycholic acid. In a separate group of 9 cirrhotic patients, fecal bile acid analysis indicated that cirrhotic patients had a significantly lower percentage of deoxycholic acid than 12 patients without liver disease; there was no significant difference in fecal lithocholic acid. The data suggest that alteration of bacterial flora and/or altered conditions for bacterial 7alpha-dehydroxylase enzyme activity in the colon could account for the virtual absence of biliary deoxycholic acid in severely cirrhotic patients.


Effects of caloric and noncaloric materials in fasting hyperbilirubinemia. Fasting hyperbilirubinemia (FHB) has previously been shown to be rapidly reversed by the ingestion of a mixed diet. This study examines the effect of carbohydrate, fat, amino acids, and noncaloric materials on FHB. After an initial fast of 15 to 39 hr, caloric and noncaloric materials were administered by mouth or vein to 13 subjects, and the total serum bilirubin (TSB) was determined frequently for 4.5 hr. Only oral glucose reversed FHB; the TSB did not change significantly from control studies after oral saline, mannitol, amino acids, and fat. In contrast, intravenous infusion of glucose resulted in a significant increase in TSB which could not be explained by osmotic or intravascular volume changes, and was nonspecific, since similar effects resulted from infusions of mannitol and amino acids. The results of this study demonstrate that oral glucose reverses FHB, and that under the conditions of these experiments other nutrients and noncaloric materials are ineffective. In contrast, intravenous glucose, mannitol, and amino acids increase the TSB by unknown mechanisms.


Presence of serum alpha-1-fetoprotein in alcoholic hepatitis. Alpha-Fetoprotein was detected in a patient with alcoholic hepatitis during the acute phase of the illness. The alpha-fetoprotein was no longer detected as clinical improvement developed. No evidence of malignant disease was found after an extensive evaluation. This case represents another example of a non-neoplastic disease associated with the presence of increased serum levels of alpha-fetoprotein.


Hereditary hemorrhagic telangiectasia: presenting as an unusual gastric lesion. A case of hereditary hemorrhagic telangiectasia, occuring in a 61-year-old male, presented as an ulcerating antral mass lesion and hepatosplenomegaly. The antral mass was demonstrated to be composed of a proliferation of blood vessels and is an unusual presentation for hereditary hemorrhagic telangiectasia of the gastrointestinal tract.


The emergency cold leg.

The emergency cold leg. The clinical state of the emergency cold leg is appropriately named. The problem may be classified as a vasospastic or venoocclusive nonocclusive lesion or as an embolic or a thrombotic occlusive lesion. Diagnosis is based on the history and results of physical examination. The status of the pulses determines the need for arteriography. Rapid recognition of the problem, with an aggressive reconstructive approach, is the cornerstone of successful management. Complications resulting from failure to carry out proper treatment or arising even when proper treatment is given may be serious and lead to loss of limb or life. The seriousness of emergency cold leg cannot be overemphasized.


Lipids in fruiting bodies of the basidiomycete Oudemansiella mucida. Dried fruiting bodies of the basidiomycete Oudemansiella mucida contain 29.1% total lipids. Their qualitative analysis revealed the presence of mono-, di-, triglycerides, sterols, free fatty acids and sterolesters. Quantitatively most significant were triglycerides (37.9%) and free fatty acids (29.7%). The phospholipid fraction contained phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine and phosphatidic acid. Gas chromatography showed the presence of a broad spectrum of fatty acids. The  ratio between the neutral and polar fractions was 6 : 1, both having linoleic acid as the main component.


Production of extracellular lysine by a strain of Bacillus coagulans. Identification and requirements for growth and lysine accumulation. A naturally deficient thiamine and methionine requiring strain of Bacillus coagulans (Ms 5) accumulates lysine in medium only when exogenous pyridoxine (optimal concentration, 0.1 mu g/ml) are supplied. Threonine exerts an inhibitory effect at higher  concentrations but pyridoxine does not.


The control of immune response in vitro by Ca2+. II. The Ca2+-dependent period during mitogenic stimulation. The Ca2+ requirement for stimulation of DNA synthesis in vitro by phytohaemagglutinin (PHA) and by lipopolysaccharide (LPS) was investigated using mouse spleen cells. During mitogenic stimulation two periods could be distinguished: a Ca2+-independent period which was followed by a Ca2+-dependent period. The results suggest that Ca2+ is required for a step preceeding DNA synthesis but not for the early initial phase of transformation.


Adoptive transfer of cell-mediated immunity to tuberculin using RNA from tuberculin-sensitive subjects. Adoptive transfer of delayed hypersensitivity skin test response to tuberculin with 'immune' RNA extracted from the sensitized lymphocytes of a healthy subject or a patient with Hodgkin's disease was successfully demonstrated in previously non-sensitive patients with neoplastic diseases including Hodgkin's disease. 'Non-immune' RNA obtained from non-sensitive man, on the other hand, failed to transfer PPD skin reactivity in non-sensitive recipients. 'Immune' RNA-mediated PPD skin test response remained positive for a considerable period of time, indicating that the effect of 'immune' RNA is systemically active. 'Immune' RNA was found to be RNase-sensitive but DNase-resistant. In vitro adoptive transfer of delayed hypersensitivity with 'immune' RNA, as assayed by lymphocyte transformation test, was unsuccessful.


Studies on the inhibition of C56-initiated lysis (reactive lysis). III. Characterization of the inhibitory activity C567-INH and its mode of action. An activity in serum which inhibits reactive lysis has recently been shown to do so by preventing the attachment of C567 complexes to cells, and hence has been designated C567-INH. This report describes certain physiochemical characteristics of the inhibitory activity. It behaves as a heat-stable pseudoglobulin, soluble in 20 per cent Na2SO4, and having alpha1 mobility on Pevikon block electrophoresis. It is excluded from CM cellulose at pH 6-0, RSC Equals 0.007 M, is retained by an XM-100 membrane and is heterogenous on Sephadex G-200, eluting in at least two peaks. The combined active materials from the Sephadex column elute from DE-52 in at least four peaks. The mechanism of action of material from each of these four peaks is shown to involve prevention of attachment of C567 complexes to membranes, and this is shown to involve an effect on C567 complexes in solution rather than an effect on the membrane. A less dramatic effect on the lysis of EC567 by limited quanities of C8 and C9 can be demonstrated. Haemolytic studies using cell-bound C567 suggest that the interaction of C567-INH with C567 involves a loose reversible association. It is therefore postulated that C567-INH inhibits reactive lysis primarily by reversibly associating with the nascent C567 complex in solution, increasing its bulk and decreasing its diffusion capacity so that it is unable to reach a cell membrane before its haemolytic potential decays.


Studies on equine immunoglobulins. IV. Immunoglobulins of the  donkey. Donkey IgGa was isolated in purified form from normal and immune donkey sera by column chromatography on DEAE-cellulose. Isolated donkey IgGa and mixtures of (IgGa+IgGb) were used as antigens to prepare rabbit reagents specific for equine IgGa or IgGb. Antibodies present in sera obtained from a single donkey at various times during the course of hyperimmunization with BSA were isolated by immuno-adsorption. The class or subclass of immunoglobulins present among isolated, donkey anti-BSA antibodies was determined by use of specific rabbit anti-equine immunoglobulin reagents. The homologues of horse IgGa, IgGb, IgGc and IgA were identified in normal donkey, mule, hinny and zebra serum.


A comparative study of anaerobic Coryneforms. Attempts to correlate their anti-tumour activity with their serological properties and ability to stimulate the lymphoreticular system. Various strains of anaerobic coryneforms and the closely related Propionibacteria have been compared in vivo with respect to their anti-tumour activity. Their effectiveness has been correlated with their serological relationship and to some extent with their ability to stimulate the lymphoreticular system. Organisms belonging to Corynebacterium acnes groups I and II and C. avidum group IV were active anti-tumour agents, although of varying effectiveness. These strains are serologically closely related and all produce a soluble cross-reacting antigen. The single C. granulosum group III strain which we tested, an unclassified coryneform, and the classical Propionibacteria did not cross-react with the main group and had little or no anti-tumour activity. At the high dose (0.7 mg) we used, all strains, whether they inhibited tumour development or not, enhanced clearance of colloidal carbon and stimulated production of an inflammatory peritoneal exudate; at lower dosage the results were too variable to permit valid comparison. At the higher dose anti-tumour activity of a strain appeared to correlate best with ability to produce splenomegally and decrease red cell volume in the blood.


Effect of a potent interferon inducer on acute and chronic lactic dehydrogenase virus viremia. The effect of a single and multiple doses of polyinosinic-polycytidylic acid (poly (I). poly(C) on lactic dehydrogenase virus viremia has been studied. A single injection of 200 mug of poly (I)-poly(C) given 6 h before infection with lactic dehydrogenase virus caused a temporary decrease in viremia of about 2.5 log10. Repeated injections of 100 mug of poly(I)-poly(C) every 24 h beginning at -24 h caused a temporary decrease in viremia of about 4.0 log 10. Therapeutic treatment with optimal dosage schedules (100 mug every 24 h) caused a reduction in viremia of 1.0 to 2.0 log10. This decrease lasted for at least 24 h after the treatment was stopped. Some possible reasons for the limited effectiveness of poly (I)-poly(C) in this system are discussed.


Immune response to hepatitis B surface antigen. A total of 69 persons were investigated for assessment of cell-mediated and humoral immunity to hepatitis B surface antigen (HBsAg). Three groups, each consisting of 20 normal persons, 20HBsAg carriers, and 20 convalescent hepatitis B patients, were studied for HBsAg, anti-HBs, and leukocyte migration inhibition with purified HBsAg. Sequential sampling if an additional group of nine acute hepatitis B patients defined the cellular and humoral immune response to HBsAg. The antigen was eliminated rapidly by mounting of cell-mediated immune response detectable for a limited period, followed by antibody response in relatively few patients moore than 3 months after clearance of circulating HBsAg.


Hepatitis B antigen in saliva, urine, and stool. A survey of hepatitis B patients, asymptomatic hepatitis B antigen (HBsAg) carriers, and control subjects was conducted to determine the relationship between antigenemia and antigen excretion in saliva, urine, and stool. Radioimmunoassay was used to detect HBsAg. Specificity-confirmed HBsAg was detected in the saliva of 6 (30%) of 20 antigenemic patients, 1 (5%) of 20 nonantigenemic patients, 14 (34%) of 41 carriers, and 0 of 112 controls. HBsAg was detected in urine only after 100-fold concentration of first-morning specimens. Specificity-confirmed HBsAg was present in the urine of 7 (16%) of 43 carriers; unconfirmed HBsAg was found in the urine of 5 (13%) of 38 patients and 5 (5%) of 112 controls. Unconfirmed HBsAg was detected in concentrated stool specimens from 5 (46%) of 11 patients and 3 of 8 carriers and controls. Longitudinally collected specimens from antigenemic subjects showed no consistent patterns of antigen excretion.


Pathogenecity of Yersinia enterocolitica for mice. A laboratory infection of Yersinia enterocolitica in mice which closely resembles the naturally acquired human infection is described Intravenous inoculation of mice with small numbers of Y. enterocolitica gives rise to a systemic, pyogenic infection involving primarily the spleen, liver, and lungs. Massive neutrophil infiltration of these organs occurs early in the infection, eventually leading to large abscesses and pulmonary consolidation. Mice infected intragastrically show neutophil infiltration in the Peyer's patches of the distal ileum less than 24h postinfection. The Peyer's patches are unable to contain the infection which spreads to the mesenteric lymph node, causing large abscesses in the medullary regions. Soon after, the infection becomes systemic with abscesses forming in the liver, spleen, and lungs, and the total peripheral leukocyte count rises dramatically to over 30,000/mm2. A serological response, in the form of agglutinating antibody, begins to appear 2 weeks after infection. Possible causes of death and the usefulness of this infectious disease model are discussed.


Coccidioidin and merthiolate in previously sensitized animals. The effect of merthiolate, which is used as a preservative in skin test materials, on skin test reactions was determined in guinea pigs. In four groups of animals, merthiolate in basal medium produced skin tests at 24 and 48 h characterized by erythema and/or induration in an intermediate region, i.e., 5 plus or minus 2.2 mm. One of the four groups of animals was a nonsensitized control group. The other three groups were subcutaneously sensitized with (i) merthiolate and saline, (ii) killed Coccidioides immitis arthrospores, and (iii) merthiolate with killed C. immitis arthrospores. Coccidioidin only and merthiolate in coccidioidin produced positive delayed results in groups 3 and 4, which were sensitized with arthrospores. A synergistic effect of merthiolate and coccidioidin was observed in animals of group 4 sensitized by merthiolate with killed C. immitis arthrospores. This effect was observed at 24 h when positive reactions of coccidioidin with merthiolate were significantly greater than skin tests with plain coccidioidin.


51Cr release microassay for measuring cell-mediated immunity in chickens. A microcytotoxic assay, based on the release of 51Cr from chicken erythrocytes by sensitized chicken lymphoid cells in the presence of the specific antigen, purified protein derivative (PPD), is described. The percentage of antigen-specific lysis was dependent on the lymphoid cell concentration, antigen concentration, time of incubation, and incubation temperature. Maximum antigen-specific lysis of nonspecific target cells was induced with a spleen-to-target cell ratio of 100:1 and with a peripheral blood lymphoid-to-target cell ratio of 1,000:1 in the presence of 100 mug of PPD per ml over a 20-h incubation period. As the concentration of PPD was increased from 25 to 200 mug/ml the specific lysis that incubation for 20 h. Incubation at 41 C produced significantly higher (P smaller than 0.05) antigen-specific cytotoxicity than incubation at 37 C.


PCA reactions with mouse antibodies in mice and rats. Mouse IgE was titrated in rats. The sensitization period was 2 h. The results were consistent and corresponded to the titers obtained in young adults SJL mice using a sensitization period of 48 h. With longer sensitization periods in rats, higher antibody titers were obtained. The optimum sensitization period in rats was found to be 48 h. Old CFW mice are inadequate for mouse IgE titration. IgG1 will not give a PCA reaction in rats. IgG1 titers are higher in SJL or A/J mice than in young CFW mice, and markedly higher than in old CFW mice.


Contact sensitivity in vitro. II. The effect of beryllium preparations on the proliferative responses of specifically allergised lymphocytes and normal lymphocytes stimulated with PHA. Beryllium in various physical forms was studied for its ability to induce increased 14C-thymidine incorporation by allergised guinea pig lymphocytes. No increase was observed. It was further shown that beryllium had an inhibitory effect on the response of normal lymphocytes to PHA and on the response of allergised lymphocytes to antigen. These results support the finding in other systems that beryllium can suppress DNA synthesis.


Specific inhibitory action of polyunsaturated fatty acids on lymphocyte transformation induced by PHA and PPD. Both saturated (SFA) and unsaturated fatty acids (UFS) inhibited PHA- and PPD-induced lymphocyte transformation. The degree of inhibition varied with the nature of the fatty acid (FA) and further comparison with their effect on unstimulated lymphocyte cultures suggests a specific action of polyunsaturated fatty acid (PUFA) on the lymphocyte PHA- and -PPD interaction. These findings are briefly discussed in the light of similar observations with PUFA-derived prostaglandins.


Nippostrongylus brasiliensis infection in rats. Both antibodies and sensitised cells are necessary for the immunological control of developing larvae. A larval N. brasiliensis infection is expelled before reaching maturity in rats given both antiserum and sensitised lymphocytes but developing larvae are not expelled from rats given antiserum or cells alone. The immunity of animals given both components is as strong as immunity in actively immunised rats. The effects of immunity do not become evident until the larvae have migrated to the intestine. Developing larvae are more susceptible to the combined action of cells and antiserum than adult worms.


Cell-mediated immunity to insulin and its polypeptide chains in insulin-treated diabectics. 35 insulin-treated diabetics and 25 control persons were studied for in vivo and in vitro manifestations of hypersensitivity to insulin and its isolated polypeptide chains. Delayed hypersensitivity reactions were determined by skin testing, and blastogenic conversion tests were done with peripheral blood lymphocytes. None of the controls and 62.9% of the patients gave positive reactions in the blastogenic test, most of these reacted to the intact molecule, fewer patients responded to B chains, and very few responded to A chains. None of the controls and only three of the patients produced positive delayed hypersensitivity reactions to insulin, and all of these were to the intact insulin molecule.


Leukocyte reactivity to alpha-gliadin in dermatitis herpetiformis and adult coeliac disease. The leukocyte adherence inhibition test has been used to study leukocyte reactivity to alpha-gliadin in 20 patients with dermatitis herpetiformis and 11 patients with adult coeliac disease. Leukocytes from both groups of patients showed a significant inhibition of leukocyte adherence thus indicating possible cell-mediated reactivity to this fraction of gluten.


Potentiating effect of D-2-O on the ascaris-induced, reagin-mediated model of asthma in the Rhesus monkey studied with a double aerosolized antigen challenge technique. Rhesus monkeys with IgE-mediated sensitivity have respiratory reactions following aerosol challenge with antigen. Studies of agents affecting this response have previously been limited by variations in animal responsiveness at different times. A method for avoiding this variability was tested in these experiments by comparing the degree of response to two antigen challenges during the same experiment and subsequently repeating the experiment with antigen dissolved in a mixture of H-2-O and D-2-O. D-2-O was selected for testing because it should theoretically potentiate histamine-mediated respiratory responses. The results of the studies indicated that there is a potentiating effect of D-2-O on the antigen-induced respiratory response in rhesus monkeys. The double antigen challenge system described should have potential use for study of a variety of inhibiting or potentiating agents on the rhesus model of asthma.


Circumvention of immunological tolerance to bovine serum albumin by diazotized antigen as a function of hapten density. Immunological tolerance was induced in adult guinea pigs by the injection of 10-100 mg of centrifuged BSA. The tolerant state was readily circumvented, in terms of both humoral antibody response and delayed hypersensitivity, by the immunization with sulfanilated BSA with different degree of conjugation. The level of anti-BSA and intensity of the skin reactions to BSA evolked in animals with circumvented tolerance responding to the challenge immunization with sulfanilated BSA were inversely related to the number of sulfanil groups per BSA molecule. On the basis of the experimental data, the mechanism of the circumvention of tolerance is discussed.


Smoking habits in Romania.

Smoking habits in Romania. The main aspects and consequences of tobacco use resulting from a series of field surveys in Romania are reviewed. The proportion of smokers in middle-aged adults is about 10 per cent for women and 40 to 50 per cent for men. Cigarettes are practically the only form of tobacco consumed--cigars are very rarely smoked and pipe-smoking accounts for less than 1 per cent of tobacco use. Less than 10 per cent of smokers use more than 25 cigarettes a day. Age at starting to smoke is low, with a quarter of smokers taking up the habit by the age of 15. Ninety per cent of the smokers admit to inhaling, and about half the men and nearly all the women smoke filter cigarettes.


Surveillance of congenital malformations and other conditions of the newborn. A brief description is given of a birth defects monitoring system in operation on a monthly basis in Norway since 1 January 1971. The system has disclosed apparent epidemic increases in congenital dislocation of hip and anomalies of the limbs, caused by changes in registration practice. An increase in the registration of anomalies of the uro-genital system, that started in the autumn of 1972, is currently being investigated in depth through a case-control study covering all cases of hypospadias and undescended testicles that are registered in the total population of births in Norway. However, this population is too small to provide a quick answer to urgent questions. To overcome this obstacle international collaboration is essential.


Associations between oral clefts and drugs taken during pregnancy. Associations between drug consumption during pregnancy and the birth of children with oral clefts were studied in material from the Finnish Register of Congenital Malformations, consisting of 599 children with clefts and their matched controls. Information concerning maternal drug consumption was partly prospective. During the first trimester, analgesic, chemotherapeutic and antineurotic drugs had all been significantly more frequently used by the mothers of children with clefts, than by the control mothers. The consumption of drugs was in general more frequent among mothers of children with cleft lip, with or without cleft palate, than among those of children with cleft palate alone.


[Suicidal problems in Norway (author's transl)]. The author gibes a review of suicide problems in Norway. The suicide rate has in Norway traditionally been low compared with the other Scandinavian countries, about one third, and has remained so for a century. In the last century the suicide rate increased with increasing age. In this century it has come to a peak in the age groups of 50-69 years for men and 50-59 years for women. There are only small differences in the suicide rate for the different parts of the country, with a slightly higher rate in the most densely populated areas. The ratio male/female is now 3:1, and has changed little. The suicidal methods are discussed, as is also the relation of suicidal acts to alcohol and drug problems. Precipitating factors are discussed, as are also vulnerable groups, among them the sailors, where the suicidal rate is about three times higher than of the average population of the same age and sex. Norwegian follow-up studies on the future fate of suicide attempters are mentioned. Finally, the author gives an outline of the present state of suicidology in Norway.


The impact of the New York State abortion law on black and white fertility in Upstate New York. The availability of abortions on demand, as provided by the new New York State abortion law, had a profound effect on fertility in a large community with previously very restricted access to illegal abortions. Improved family planning and the availability of abbortions as a backup measure combines with a preference for a smaller family size led to a one-third reduction of live births in the short period since the new law became effective. Planning for paediatric and obstetric care had to be adjusted accordingly. The frequently hypothesized or reported secondary effects on illegitimacy or infant mortality could not be observed. The trend towards increasing illegitimacy was slowed down, but not reversed. Infant mortality continued its short-term erratic pattern; the trend towards a higher proportion of teenage pregnancies cancelled the effect that pregnancies at higher risk for infant death are also at higher risk for abortion.


The validity of data from next-of-kin in studies of mortality among migrants. The design of the British-Norwegian Migrant Study afforded a unique opportunity for comparing in a large sample certain items of information on the same individual, as reported by himself some time before death, with the responses on the same points given after his death in a questionnaire addressed to his next-of-kin. This paper describes a comparison of the degree of agreement between these two methods of obtaining data and draws attention to possible sources and directions of bias involved in the use of information supplied after death by the next-of-kin. Validity, as measured by agreement with the subject's own replies, depends in part on the topic and nature of the information required and on the population included in the survey. A built-in check, such as that incorporated in the British-Norwegian Study, confers the advantage that such biases can be identified and the numerical effect of adjusting for them assessed with some confidence.


Cancer of the oesophagus in Brittany: an incidence study in Ille-et-Vilaine. Mortality from cancer of the oesophagus is known to be high in Brittany and Normandy. A morbidity survey was initiated to confirm this finding. A registry was set up in Rennes for the département of Ille-et-Vilaine. In the course of six years (1968-73) 718 cases were entered, 669 males and 49 females; the annual incidence rate for males is 29-4 per 100,000, one of the highest ever described. Most tumors are in the middle third of the oesophagus (49 per cent); 33 per cent are in the lower third. Most tumors were squamous cell carcinomas; 7 per cent were adenocarcinomas. There is an area of high endemicity in the rural areas of the north of Ille-et-Vilaine with rates reaching 60 per 100,000. The relationship with drinking habits is currently being investigated.


Amniocentesis: indications and complications. Transabdominal amniocentesis is described and our experience with this method is presented. No severe maternal complications were recorded. In two cases accidental puncture of fetal blood vessels in pregnancies over 40 weeks led to delivery by cesarean section. We believe that, although fetal complications are rare, the indications for amniocentesis should be significant enough to discount the hazards of the procedure.


Clinical study of intersex states in Nigerian children. Early diagnosis of intersex is essential to avoid indecision and to establish the correct sex of rearing before mistakes have been made. Diagnosis of adrenogenital syndrome is singularly important for delay in diagnosis may lead to stunted growth or death from electrolyte imbalance. All others except the hormonally induced types are sterile, the gonad can be ignored and the sex of rearing is determined by the external genital configuration which can be most easily created by surgery plus hormones. Patients except those with maternal virilization may also need castration and hormone therapy corresponding with the chosen sex of rearing.


Incidental appendectomy in vaginal surgery. We have analyzed 225 cases of incidental appendectomy performed during 2200 vaginal operations over a six-year period. The main vaginal procedures were 1700 hysterectomies and 500 colpotomies for tubal ligation, ovarian cystectomy and diagnostic procedures. Other than prolonging the operating time by about 15 minutes, there was no sign of increased morbidity or duration of hospital stay.


Out-patient uterine vacuum curettage without anesthesia. The Vabra aspirator was used for vacuum curettage in 130 Nigerian women needing uterine curettage. The procedure was performed on an out-patient basis without anesthesia. Eighty-nine patients (68.5 percent) had their operations at the time of their first visit to the gynecological consultative clinic. The procedure was acceptable to all the patients; 117 (90 per cent) reported no pain and all 120 patients seen at the follow-up clinic returned to work a few hours after the procedure. The vacuum curettings were adequate for histological diagnosis in 92.3 per cent of the patients in whom specimens were obtained. There was no immediate or long-term complications.


A new, rapid fluorometric determination of retinol in serum. A new procedure for the determination of retinol (vitamin A alcohol) in blood has been devised. It consists of direct measurement of the fluorescence of retinol (excitation 335 nm.; emission 458 nm.) in diluted serum, and is considerably more rapid than any previous method. Such measurements are feasible because the intensity of fluorescence of retinol bound to its transport protein (retinol-binding protein) is sufficiently high that other natural occurring fluorescent substances in blood do not interfere significantly. When the new method was compared to a conventional procedure employing extraction and chromatographic separation of retinol on columns of alumina, the correlation coefficient was 0.85.


A rapid method for measuring miotic activity of drugs in the intact mouse eye. A rapid and precise method for evaluating the miotic activity of cholinergic drugs has been developed based on Long's method for measuring the rate of mydriasis. The rate of reversal of mydriasis developed previously in the intact mouse eye by a mild mydriatic (phenycyclidine) is used to evaluate the miotic activity. The method provides a useful tool for measuring and comparing the miotic activity of acetylcholine agonists and cholinesterase inhibitors.


Pathways for the response of the eye to injury. The role of a neural pathway in the disruption of the blood-aqueous barrier of the rabbit after two irritating stimuli, topical nitrogen mustard, and paracentesis, was studied. Retrobulbar anesthesia or section of the ophthalmic division of the trigeminal nerve decreased the breakdown of the blood-aqueous barrier, as measured by protein in the aqueous humor, after topical nitrogen mustard. Sensory denervation, accomplished with retrobulbar alcohol, virtually prevented the protein rise in the aqueous humor. Disruption of the blood-aqueous barrier after paracentesis, however, was not affected by retrobulbar anesthesia or alcohol denervation. Therefore, the increased protein in the aqueous humor after an irritative stimulus appears to be mediated by at least two pathways. The response to a stimulus such as nitrogen mustard depends on sensory innervation and is not mediated by prostaglandins to any important degree. The response to paracentesis does not require sensory innervation and appears to be mediated, at least in part, by prostaglandins.


Experimental tonography on enucleated human eyes. I. The validity of Grant's tonography formula. When newer tables for parameters in Grant's formula for tonography are used, one obtains far higher values for the outflow facility than those generally accepted. The aim of this iinvestigation has, therefore, been to test the validity of the tonography formula. The outflow facility determined by constant-rate perfusion in enucleated eyes has been compared with the results of tonography on the same eyes in the steady-state condition. Loss of intraocular fluid during tonography has been measured directly by means of a new technique. Outflow facilities determined according to the two methods show very good agreement. Thus, when the production of aqueous is held at a constant rate, and when there is no change in intraocular blood volume. Grant's tonography formula is found valid, provided absolute values for Po, Pt, and dV are used.


Cone dark adaptation: the influence of halothane anesthesia. Cone dark adaptation (da) (measured with an electroretinogram [ERG] method) is found to be severely retarded when the subject (human and macaque monkey) is under halothane anesthesia. The degree of retardation depends both on the halothane level and on the bleach history. The effect of bleach history shows a great similarity with psychophysical findings concerning foveal da. Interpretation of halothane effects, therefore, is possible in terms of an existing model on cone pigment kinetics.


Evidence for lens oxoprolinase, an enzyme of the gamma-glutamyl cycle. The presence of oxoprolinase, an enzyme of the gamma-glutamyl cycle, not previously reported in the lens was demonstrated by organ-culture technique and from a study of the partially purified enzyme. The evidence for oxoprolinase in intact rabbit lens is based on the following: (1) [14C]-labeled oxoproline is utilized by the lens giving rise to labeled CO2, (2) [14C]-oxoproline is converted to glutamic acid, which is subsequently incorporated into glutathione, (3) formation of labeled glutamic acid and CO2 from [14C]-oxoproline is effectively blocked by a structural analog of the compound L-2-imidazolidone-4-carboxylic acid, a known inhibitor of oxoprolinase. The enzyme was partially purified from bovine lens capsule epithelium and certain of its properties were examined. Ocular lens was also found to contain significant amounts of oxoproline, an intermediate of the gamma-glutamyl cycle.


Blood pressure and pressure amaurosis.

Blood pressure and pressure amaurosis. Susceptibility to pressure amaurosis was measured in young research subjects before and during blood pressure elevation induced by intravenous infusions of phenylephrine. Intraocular pressure elevations were produced by paralimbal suction; we measured the highest level to which intraocular pressure could be raised without obliterating perception of a slowly flickering stimulus in the nasal field of vision. Elevation of systemic blood pressure was accompanied in all subjects by a corresponding increase in the highest "safe" level of intraocular pressure. This observation confirms the commonly held hypothesis that pressure amaurosis is the result of pressure-induced neuroretinal ischemia.


Lymphocyte-induced vitreous membranes: a comparative study with leukocyte- and platelet-induced vitreous membranes. Vitreous membranes were induced in rabbit eyes by injecting an autogenous lymphocyte preparation. These membranes were compared with membranes induced by autogenous leukocyte and platelet preparations. Lymphocytes gave rise to faint, nonprogressive membranes, while leukocytes and platelets produced dense, long-standing membranes. It is suggested that lymphocytes may not be the cause of this weak response, as other cells in the preparation may be involved.


Effects of antibiotics on the in vitro rabbit corneal endothelium. The effects of ten antibiotics and two preservatives on the rabbit corneal endothelium were studied by in vitro perfusion. Dose-related effects on corneal thickness were seen with certain antibiotics. No correlation was found between structure, origin, or bacterial sensitivity of the antibiotics and their effect on corneal transplants. Five distinct morphological changes were observed: two were transient and only one was specific. Although clinical dose correlation was not intended, further investigation is needed on erythromycin, bacitracin, chloramphenicol, penicillin, oxacillin, and cephalothin.


Anti-inflammatory effectiveness of topically administered corticosteroids in the cornea without epithelium. The present studies demonstrate that modification of the derivative of a given steriod base alters its anti-inflammatory potential as measured by suppression of leukocyte invasion of the cornea. A comparison of each drug's corneal bioavailability with its anti-inflammatory effectiveness shows the acetate derivative of prednisolone to be a more potent anti-inflammatory agent than the phosphate derivative. Similarly, the free alcohol derivative of dexamethasone proved to be more potent than the phosphate derivative. Increasing the concentration of prednisolone acetate from 0.125 per cent to 1.0 per cent results in a significant increase in its anti-inflammatory effectiveness in the cornea following topical administration. The same increase in prednisolone phosphate concentration does not produce a significant increase in its ability to suppress polymorphonuclear leukocyte infiltration of the cornea. When the epithelium of the inflamed cornea is intact, prednisolone acetate, 1.0 per cent ophthalmic suspension, is the most effective of the corticosteroid preparations studied. In the absence of an intact epithelium, prednisolone acetate, 1.0 per cent ophthalmic suspension, again produces the greatest mean reduction in polymorphonuclear leukocytic infiltration of the cornea although here one cannot demonstrate a statistically significant difference from the anti-inflammatory effect produced by prednisolone phosphate, 1.0 per cent ophthalmic solution, or dexamethasone alcohol, 0.1 per cent ophthalmic suspension. Overall, therefore, prednisolone acetate 1.0 per cent is the most effective of the topical agents studied for suppression of corneal inflammation.


The effect of tear substitutes on tear film break-up time. Twelve commercial artificial tear solutions and a newly developed one were evaluated as to their effect on tear film breakup time (BUT) in ten normal subjects. Instillation of one drop of these solutions altered the BUT in such a way that serial BUT measurements could be used as an index of retention time. Results demonstrated significantly longer retention time for three related products (Adapt, Adapette, and Adsorbotear) and a newly developed product (Alcon 0413) [Tears Naturale (Alcon TM)]. This method appears to be an accurate nonirritative way of assessing retention time of tear substitute/vehicles and demonstrate values much longer than previously reported by other methods.


Radiographically determined lung volumes at full inspiration and during dynamic forced expiration in normal subjects. Posteroanterior and lateral radiographs of the lungs of 30 normal adults were obtained at total lung capacity (TLC) and 1 sec after initiation of forced expiration from ttlc. simultaneous spirographic recording of the volume expired (FEV-1) and independent plethysmographic recording of TLC were obtained. Two types of measurements were made from the radiographs: 1) surface areas of the lung fields, measured planimetrically; and 2) combinations of linear distances between selected points. These radiographic measurements and the measured lung volumes were subjected to computer analysis to develop reliable formulas for determining lung volumes solely from radiographs. Formulas derived using planimetry produced multiple correlation coefficients of 0.95 for TLC and 0.89 for volume during forced expiration. Use of linear measurements produced multiple correlation coefficients of 0.92 for TLC and 0.84 for dynamic volume. Thus, in normal subjects, static lung volumes and lung volumes during dynamic forced expiration can be determined from radiographs.


Roentgenographic study of blood flow redistribution in acute pulmonary edema in dogs. Acute pulmonary edema was induced in anesthetized vertically suspended dogs by inflating a ballon catheter in the descending thoracic aorta and infusing saline. Indices measured included pulmonary arterial pressure, left ventricle end-diastolic pressure, and cardiac output. Pulmonary blood-flow distribution was determined in frozen tissue after injecting radioactive macroaggregates labelled with 125-I, 131-I, and 99m-Tc. Randomized serial chest roentgenograms were interpreted by three radiologists not taking part in the physiologic studies. When left ventricular end-diastolic pressure was raised initially, roentgenograms usually revealed only interstitial edema. As alveolar edema increased, blood-flow to the lung bases decreased as measured physiologically. The roentgenographic and physiologic data show that in acute experiments on normal lungs, blood-flow redistribution occurs in association with alveolar edema rather than interstitial edema.


Controlled deposition of tantalum powder in a cast of the human airways: applications for aerosol bronchography. A hollow latex cast of the human larynx and tracheobronchial tree extending to 2 mm diameter airways "inhaled" tantalum powder (mass median aero-dynamic diameter equals 9.2 mum, omicron-g equals 1.41) at 8 liters/min. Tantalum deposited within the cast as predicted by preliminary deposition  calculations. These calculations predicted deposition surface densities among greater than 2 mm diam. airways to have a range of similar to 2:1, and also predicted less than 5% alveolar deposition. A deposition surface density of 8 mg/cm-2 provided good bronchographic visualization. Single rice grains located within some airways were distinctly outlined. The small amount of tantalum needed to outline the airways be a simulated voluntary inhalation indicates that tantalum may be suitable for use as a bronchographic contrast medium when administered under strictly controlled exposure conditions.


Myocardial wall motion evaluation using amplitude frequency spectral analysis. Frequency spectral analysis of cardiac wall motion as calculated from the recorded video kymogram are demonstrated. Characteristic harmonics following the fundamental frequency of the cardiac rate have been demonstrated to have an alternating sequential energy distribution as measured from the free margin of the left ventricle of humans. The characteristic patterns occur in approximately 80% of the 50 patients evaluated. The correlation of these harmonics with physiologic events is not yet complete. Amplitude frequency spectral analysis offers an objective method for analysis of cardiac kymography not otherwise possible.


Subacute sclerosing panencephalitis: incidence among ethic groups in Israel. During the period 1968-73, 46 patients with subacute sclerosing panencephalitis (SSPE), a fatal childhood disease related to rubeola, were encountered in Israel. The incidence per million population was 3.4 for Sephardic Jews, 3.2 for Arabs and only 0.5 for Ashkenazic Jews. An environmental factor which sharply demarcated Arabs and Sephardic Jews from Ashkenazic Jews, and which might account for the differences in incidence of SSPE, was family size: 42.3% of Arab families and 21.5% of the families of Sephardic Jews but only 1.1% of Ashkenazic Jewish families had five or more children. It is postulated that older siblings in large families might constitute vectors which introduce rubeola to the younger siblings at a time when they are at unusual risk of developing SSPE (i.e. before two years of age), whereas in small families, rubeola tends to be acquired later in childhood when the risk of SSPE is reduced.


Conservative vs. surgical treatment of vesicoureteral reflux. The treatment of choice in vesicoureteral reflux in children is discussed on the basis of experience with 160 refluxing ureters. Two-thirds of the ureters responded well to conservative nonsurgical treatment and the majority of these are now completely cured. Thirty-three ureters, not responding to conservative measures, were treated surgically by the Leadbetter-Politano or Bischoff methods. It is concluded that 1) conservative treatment should be attempted before recourse to surgery and 2) Bischoff's technique should be given first consideration in cases with dilated and poorly functioning ureters.


Dynamic radiography. An evaluation of cardiac motion by the analysis of scattered radiation during fluoroscopy. A non-invasive technique, utilizing scattered radiation, has been developed to monitor and measure the motion of the epicardial surface during fluoroscopy. A number of dogs were studied using a monitoring device which consists of an X-Ray beam collimator, collimated sodium iodide dectetors and their associated electronics. The detector signals are computer processed to obtain frequency fingerprints of epicardial motion that may, in the future, indicate the condition of the myocardium.


A case of maternal hyperparathyroidism presenting as neonatal tetany. A three-week-old infant presented with tetany and with serum calcium and phosphate levels compatible with the diagnosis of hypoparathyroidism. Investigation of the asymptomatic mother revealed serum calcium and phosphate levels and urinary phosphate clearance values diagnostic of hyperparathyroidism. The maternal hyperparathyroidism was treated by removal of a parathyroid adenoma.


Toxicity of metrizamide and meglumine iocarmate in the spinal subarachnoid space. An experimental study in rats with special reference to long-term effects. The toxic effects of metrizamide, a new non-ionic contrast medium, and of meglumine iocarmate (Dimer-X), were investigated after injection into the spinal subarachnoid space of rats. Convulsions were seen in 8 of 11 animals with meglumine iocarmate. No such irritative effects seen with metrizamide. Histologic examiniation of the spinal cord was carried out here 4 months. No changes were seen which could be ascribed to toxic effects of the contrast media.


Bilateral hearing loss as a sequel to unilateral acoustic trauma. Within the framework of a study on the natural history of acoustic trauma, over 600 soldiers were examined. A higher rate of bilateral acoustic trauma was observed among soldiers with longer service. Comparison between unilateral and bilateral acoustic trauma showed a greater severity of damage in both ears of the latter group. Comparison of the progression of the damage in the same persons on two consecutive examinations suggested the same trend. It is suggested that bilateral acoustic trauma may be a later and more severe stage in some types of noise-induced damage.


Renal arteriography. The choice of contrast material. Angiographers responding to a questionnaire disagreed on the agent of choice for selective renal arteriography. Opinion differed regarding the advantages of sodium versus meglumine salts, diatrizoate versus iothalamate, and 50-60 percent concentration agents versus those in the 70-76 percent range. Although an increasing number of angiographers are using more concentrated agents for renal arteriography, there is little information available to indicate their safety for this purpose. Dog experiments were done to see whether Renografin 76 percent (R-76), at a reasonable dose, caused more severe changes than the lower concentration agents. The studies failed to demonstrate significant differences between R-76 and the less concentrated agents indicating that R-76 can be used for selective renal arteriography if the additional iodine is deemed necessary. Factors affecting nephrotoxicity of contrast media in patients with normal and abnormal kidneys are discussed.


The direct measurement of cytochrome P450 in unfixed tissue sections. A method for the measurement of cytochrome P-450 in unfixed cryostat sections is described. The sections are incubated for 10 minutes at room temperature in a buffered solution containing polyvinyl alcohol and sodium dithionite. Two incubations are performed on serial sections, one in nitrogen and the other in carbon monoxide. Readings are taken on a Vickers M85 microdensitometer fitted with a high sensitivity photomultiplier amplifier system, the measurements being made on corresponding fields in the serial sections. Subtraction of the nitrogen values from the carbon monoxide values, after allowing for an absorption shift, gives the absolute spectrum of cytochrome P450. The subtraction corrects for the tissue content of other haem-containing proteins. The cytochrome P450 spectrum shows a sharp maximum at 450 nm, and two other minor components absorbing at 444 nm and 458 nm. The content of cytochrome P450 in animals fed with phenobarbitone was 2.4 times greater than in control animals.


Methylation or lactonization of acid mucins by acid methanolic reagents in histochemistry. With the demonstration that cationic dye staining of acid mucins can be prevented by treatment with hydrocholoric acid or thionyl chloride in nonpolar, nonalcoholic solvents it has been contended that the blockade has occurred by lactonization of the acid mucosaccharides. It is further contended that also in methanolic HCl lactonization is the only process by which cationic dye staining of acid mucins occurs to the exclusion of methyl esterification of the carboxyls. We have demonstrated that an acetylation adequate to prevent the PAS reaction of mucins does not prevent either direct cationic dye staining or its blockade by methylation, and that periodic acid cleavage of the 2,3 glycol of hyaluronic acid mucins does not prevent methylation blockade of cationic dye staining. With occupying or destroying the variation of(3) hydroxyl on which variation of-lactonization would have to occur it is believed that the successful blockade must have occurred by methyl esterification of the carboxyl. Lactonization must be regarded as an alternative rather than an exclusive pathway for this blockade.


[On the lysosomal pattern in the kidney of normal rats with special reference to the protein excretion(author's transl)]. Using the histochemical reaction for acid phosphatase the lysosomal pattern of the proximal renal tubule has been investigated in virginal female rats, in the course of pregnancy, and in females following pretreatment with progesterone. Furthermore the stage of gestation, phases of sexual cycle, the urinary output as well its protein concentration and the protein excretion have been taken into account. From the metabolic studies it becomes clear, that the urine values depend on the sexual phase and the duration of gravidity. On the basis of our histochemical results strong evidence occurs, that the lysosomes in the proximal renal tubule are influenced by both, the proteins in the primary urine and by sex hormones. In all collectives investigated an increase of the protein concentrations in the urine runs always parallel to an increase of the number of small and medium-sized lysosomes; the big ones disappear. At the end of pregnancy and after pretreatment with progesterone acid phosphatase seems to diffuse out of the lysosomes in the epithelial cells of the S1- and S2-segments.


Evaluation of tear formation in the dog, using a modification of the Schirmer tear test. The Schirmer tear test for the measurement of tear formation in the dog was done according to 2 procedures: Schirmer I (the conventional procedure) and SCHIRMER II (with topical anesthesia and drying of the ventral conjunctival fornix. Results from 97 normal dogs averaged 21.0 mm wetting/minute (standard deviation (sd) plus or minus 4.2 mm) for the Schirmer I test and 11.6 mm  wetting/minute (sd plus or minus 6.1 mm) for the Schirmer II test. When atropine was injected subcutaneously in 50 dogs (0.02 mg/kg, wetting/minute averaged 9.36 mm (Schirmer I) AND 4.32 MM (Schirmer II). Two dogs in each of 3 groups had either the lacrimal gland, gland of the membrane nictitans, or both excised. Changes in tear formation were evaluated by Schirmer I and II tear tests, as well as after parenteral administration of atropine. The lacrimal gland, the gland of the membrama mocotams. amd tje accessory lacrimal glands and mucous cells were found to contribute 6.17, 35.2, and 3.1%, respectively, to tear formation.


Spinal cord compression caused by osteocartilaginous exostoses of the spine in two dogs. Two dogs, one 4 months old and the other 1 year old, had histories of progressive posterior weakness. In both cases, there was severe asymmetric proprioceptive loss but appreciable preservation of strength. Benign, cystic expansile lesions involving the facets and lamina of the thoracic spine were observed radiographically; a single lesioons was demonstrated in the other. Examination of biopsy speciments taken at the time of laminectomy revealed the lesions to be osteocartilaginous exostoses (osteochondromas, ecchondromas, exostoses). The progress of dogs 1 and 2 has been monitored for 1 1/2 years and 9 months, respectively. Dog 1 has no weakness or proprioceptive loss; dog 2 has a mild proprioceptive loss in the right hindlimb.


Acute pancreatic hemorrhage associated with iatrogenic hypercalcemia in a dog. A 2-year-old female dog with idiopathic hypolcalcemia was given intermittent calcium therapy, to effect. Excessive calcium may have led to the development of acute pancreatic hemorrhage.


Calcifying aponeurotic fibroma in a dog. A calcifying aponeurotic fibroma involving the soft tissue and bone of the occipital area compressed and attenuated the cerebellum and brainstem of a 9-year-old female dog of mixed breeding. Clinical signs included a head tilt to the left; gait ataxia; anisocoria, with a midly dilated left pupil; horizontal nystagmus, with the fast phase to the right; and a mild depression of the level of consciousness. The signs were sequential and progressive.


Perianal adenoma of apocrine origin in a dog. At necropsy, a 15-year-old male Standard Poodle had a 6-cm diameter mass in the perianal region. Histologic examination revealed an adenoma of apocrine tubular gland origin, rather than the more common type of perianal gland adenoma derived from modified sebaceous glands. The diameter of the mass had increased from 3 to 6 cm in 12 months and it had failed to respond to treatment with repository stilbestrol.


Genetic recombination in Nocardia mediterranei. A system of genetic recombination in Nocardia mediterranei ATCC 13685 is described. This strain produces a mixture of several rifamycin antibiotics. Using haploid recombinant selection and analysis procedures similar to those applied to Streptomyces coelicolor A3(2), 14 auxotrophic markers and 1 streptomycin resistance marker were located on a circular linkage map. The linkage map of N. mediterranei seems to be similar to that of S. coelicolor A3(2).


Characterization of the lipids of mesosomal vesicles and plasma membranes from Staphylococcus aureus. Mesosomal vesicles and plasma membranes were isolated from Staphylococcus aureus ATCC 6538P by protoplasting and differential centrifugation. The lipids of each of the two membrane fractions were extracted with pyridine-acetic acid-N-butanol, and the nonlipid contaminants were removed by Sephadex treatment. The lipids were then separated by passage through diethylaminoethyl-cellulose columns and characterized by thin-layer chromatographic, chemical, and spectral analyses. The lipids were separated into four discrete diethylaminoethyl fractions: (i) vitamin K2, carotenoids, C55 isoprenoid alcohol, and monoglucosyl diglyceride; (ii) cardiolipin, carotenoids, phosphatidyl glycerol, diglucosyl diglyceride, and an unidentified ninhydrin-positive component; (iii) cardiolipid and phosphatidyl glyderol; (iv) cardiolipin, phosphatidyl glycerol, and phosphatidyl glucose. Qualitatively, no difference in lipid composition between mesosomal vesicles and plasma membranes was found. However, based on equal dry weights of membrane materials, a relative quantitative difference in the amount of specific lipids in mesosomal vesicles and plasma membranes was observed. There are 4 times more monoglucosyl diglyceride, 2.6 times more diglucosyl diglyceride, 3.8 times more phosphatidyl glucose, 2 times more carotenoids, and 2 times more vitamin K2 found in mesosomal vesicles than in plasma membranes. The concentration of cardiolipin and phosphatidyl glycerol is 3.6 and 6 times greater, respectively, in mesosomal vesicles.


Sensitivity of a Bacteroides melaninogenicus strain to monosaccharides: effect on enzyme induction. The inhibition of growth in Bacteroides melaninogenicus by sugars in described. Monosaccharides such as D-glucose, D-galactose, D-mannose, and D-fructose are inhibitory at low concentrations, whereas the disaccharides sucrose and lactose are not inhibitory even at high concentrations. The major inhibitory effect of the sugar is found during the transition of lag to logarithmic growth phases. There was no primary effect of D-glucose on protein, ribonucleic acid, or deoxyribonucleic acid synthesis on cells in transition from lag to logarithmic growth. However, the addition of glucose or galactose completely abolished the induction of 3-ketodihydrosphingosine synthetase by vitamin K in vitamin K-depleted cells. Futhermore, in cells which were not vitamin K depleted, the level of this enzyme was drastically reduced by the addition of the sugar. Cyclic adenosine 5-monophosphate was unable to reverse the growth inhibition produced by glucose. In actively growing cultures, addition of sugar slows the growth rate. In these experiments the level of 3-ketodihydrosphingosine synthetase fell only after the cells had assumed the slower rate of growth. There were two indications that D-galactose was more inhibitory than D-glucose; in the presence of 0.1% D-galactose cells in lag phase did not show the increase in turbidity found in similar cells placed in medium with 0.1% D-glucose, and also D-galactose caused a greater decrease in the growth rate of actively growing cultures than was found with D-glucose. These studies suggest that the inhibitory effect of monosaccharides in lag leads to logarithmic growth transition can be ascribed to an effect on enzyme induction. On the other hand, the ability of many monosaccharides to inhibit growth, and the greater inhibitory property of D-galactose compared with D-glucose, suggests that other mechanisms may be operative as well.


Kinetics of Na+-dependent K+ ion transport in a marine pseudomonad. The effect of external Na plus concentration on the transport of K plus was studied using K plus-depleted cells of a marine pseudomonad. K plus transport was found to be a saturable process and requires Na plus. The initial rates for K plus transport over a range of external K plus concentrations were measured in suspensions containing various fixed concentrations of Na plus. Reciprocals of the initial rates for K plus transport were plotted against reciprocals of the external concentration of K plus or Na plus to yield two primary Lineweaver-Burk plots. The experimental data were found to fit bisubstrate enzyme kinetics, with a sequential type mechanism. However, the initial rate data did not allow distinction between ordered or random mechanisms. The results suggest that Na plus and K plus form a ternary complex with a specific K plus carrier molecule on the outer surface of the membrane prior to translocation and the release of K plus inside the cell.


Lipid metabolism during bacterial growth, sporulation, and germination: kinetics of fatty acid and macromolecular synthesis during spore germination and outgrowth of Bacillus thuringiensis. The timing and kinetics of fatty acid synthesis are delineated for Bacillus thuringiensis spore germination and outgrowth by analyzing [U-14C]acetate and [2-3H]glycerol incorporation into chloroform-methanol-extractable and trichloroacetic acid-precipitable lipids. In addition to measurement of pulsed and continuous labeling of fatty acids, monitoring the incorporation of radioactive phenylalanine, thymidine, and uridine from the onset of germination through first cell division provides a profile of biochemical activities related to membrane differentiation and cellular development. Upon germination, ribonucleic acid synthesis is initiated, immediately followed by rapid and extensive fatty acid synthesis that in turn precedes protein, deoxyribonucleic acid and triglyceride synthesis. Significantly, formation of fatty acids from acetate exhibits further developmental periodicity in which a large transient increase in fatty acid synthetic activity coincides with the approach of cell division. Radiorespirometric analyses indicates only slight oxidative decarboxylation of acetate and corroborates the extreme involvement of acetate in specific fatty acid biosynthetic reactions throughout cellular modification. These findings graphically demonstrate an intimate association of fatty acid metabolism with commitment to spore outgrowth and subsequent cell division.


Tryptophan biosynthetic pathway in the Enterobacteriaceae: some physical properties of the enzymes. Several physical properties of the first four enzymatic activities of the tryptophan pathway were examined using gel filtration and ion exchange chromatography. Five different patterns were noted. Differences in the anthranilate synthetase (AS) and phosphoribosylanthranilate transferase (PRT) defined these patterns. In all the organisms studied phosphoribosylanthranilate isomerase and indoleglycerol phosphate synthetase co-eluted from both diethylaminoethyl-cellulose and G-200 and thus probably are contained in a single polypeptide of 50,000 daltons. An AS-PRT complex was found in Citrobacter species, Enterobacter cloacae, and Erwinia dissolvens. In all the other bacteria examined AS and PTR were separate molecules. In Serratia marcescens, S. marinorubra, and Enterobacter liquefaciens, AS was 140,000 daltons and PRT was 45,000 daltons. In Erwinia carotavora and Enterobacter hafniae the AS was the same size as the Serratia species but the PRT was larger at 67,000 daltons. Two Proteus species had an AS and PRT of the same size as E. carotavora and E. halfniae but the Proteus AS was different in that it partially dissociated upon gel filtration. Aeromonas formicans was unique in its possession of an AS with a molecular weight of 220,000. The PRT of A. formicans was found to elute at 67,000 daltons. Possible paths of evolution of the tryptophan enzymes are discussed in terms of the results of this study. The results presented here are also considered with respect to existing taxonomic schemes of the enteric bacteria.


Comparative analysis of the lipids of Acinetobacter species grown on hexadecane. A comparative analysis of the cellular and extracellular lipids of Acinetobacter species HO1-N indicated basic physiological differences in hexadecane-grown cells. The cellular lipids obtained from hexadecane-grown cells were characterized by 3- and 18-fold increases in the phospholipid fraction and the mono- and diglyceride fraction, respectively, over that obtained from nutrient broth-yeast extract-grown cells. The cellular-associated pools of hexadecane were shown to comprise approximately 8% of the dry cell weight of hexadecane-grown cells. The extracellular lipids obtained from the culture broths of hexadecane-grown cells were comprised of triglyceride, mono- and diglyceride, free fatty acid, and wax ester. These lipids were either absent or present in minor concentrations in the culture broths of nutrient broth-yeast extract-grown cells. The exponential growth of Acinetobacter sp. on hexadecane was characterized by the significant accumulation of free fatty acid, monoglyceride, and diglyceride in the culture medium. Wax ester was shown to represent a minor portion of the extracellular lipids during the exponential growth phase, appearing in significant proportion only after the culture had entered the stationary phase of growth.


Regulation of arylsulfatase synthesis by sulfur compounds in Klebsiella aerogenes. In Klebsiella aerogenes, arylsulfatase synthesis was repressed by inorganic sulfate, sulfite, sulfide, thiosulfate, and cysteine, but not by methionine under normal growth conditions. We isolated cysteine-requiring mutants (Cys minus), and mutants (AtsS minus, AtsR minus) in which the regulation of arylsulfatase synthesis was altered. In the cysteine auxotroph, enzyme synthesis was also repressed by inorganic sulfate or cysteine. Kinetic studies on mutants of the cysteine auxotroph showed that inorganic sulfate repressed arylsulfatase synthesis and that this was not due to cysteine formed by reduction of sulfate. Arylsulfatase synthesis in the AtsS minus mutant was not repressed by inorganic sulfate but was repressed by cysteine. This mutant strain had a normal level of inorganic sulfate transport. Another mutant strain, defective in the inorganic sulfate transport system, synthesized arylsulfatase in the presence of inorganic sulfate but not in the presence of cysteine. The AtsS minus mutant could synthesize the enzyme in the presence of inorganic sulfate but not cysteine. The AtsR minus mutant could synthesize the enzyme in the presence of either inorganic sulfate or cysteine. These results suggest that there are two independent functional corepressors of arylsulfatase synthesis in K. aerogenes.


Ultrastructure of Chlamydomonas eugametos palmelloids induced by chloroplatinic acid treatment. Palmelloids induced in the unicellular green alga Chlamydomonas eugametos by chloroplatinic acid treatment have been studied electron microscopically. Thin-sectioned specimens revealed the multilayer nature of the cell walls after second division within the palmelloid. Although synchrony in cell division is lost, to a certain degree, within the palmelloid, the cells themselves appeared normal, and the presence of normal flagellar structure was confirmed. The presence of the eyespot was observed at the optical level as well as in the freeze-etched specimens. The above results support the hypothesis that the palmelloid condition of Chlamydomonas eugametos induced by chloroplatinic acid is due to an abnormality in cell wall formation rather than flagellar malfunction or loss.


Energetics of Bacillus stearothermophilus growth: molar growth yield and temperature effects on growth efficiency. The major growth yield of a prototrophic strain of Bacillus stearothermophilus under aerobic conditions on salts medium containing ammonium nitrate as the nitrogen source and glucose or succinate as the carbon source was maximal at the lowest growth temperature employed and decreased steadily as the temperature was raised. The temperature optima for growth yield and for growth rate were thus different. The molar growth yield values of the thermophile, especially at the lower growth temperatures, were similar to those reported for aerobically grown mesophilic bacteria, both on glucose and on succinate. At the higher growth temperatures, a lower proportion of glucose carbon was incorporated into cells and a correspondingly greater proportion was left incompletely utilized in the medium, mostly as acetate. This suggests a greater inefficiency in the coordination of the nonoxidative and oxidative phases of glucose metabolism at the gigher temperatures. Another factor causing a decreased cell yield at higher temperatures was possibly an uncoupling of energy production from respiration. The rates of respiration by intact cells of the thermophile on glucose and on succinate followed the Arrhenius relationship from 55 C to 20 C, which is some 20 C below the minimal growth temperature of the organism. The Arrhenius constant was 17.1 kcal/mol for glucose oxidation and 13.5 kcal/mol for succinate oxidation. These results are comparable to those reported for some mesophiles, and they suggest that the inability of the thermophile to grow at temperatures below about 41 C is not due to an abnormally high temperature coefficient for the uptake and oxidation of the carbon source.


Relationship of macromolecular synthesis to competence induction in a group H streptococcus. Group H streptococcus strain Wicky, which was induced to competence for genetic transformation with competence factor (CF) derived from a related strain, displayed reduced rates of ribonucleic acid (RNA) and peptidoglycan synthesis. Pulse-labeling studies revealed that the inhibition of both RNA and peptidoglycan synthesis was maximal at the peak of competence and decreased as competence declined. These studies indicated that competence induction had only a slight effect on the rate of protein synthesis. Trypsin inactivation of CF prevented the reductions in synthesis normally elicited by CF preparations. If the addition of trypsin was delayed until 5 min after the addition of CF, competence induction and decreased synthesis of RNA and peptidoglycan were again apparent. Thus, the alterations in the synthesis of these macromolecules appeared to be related to the induction of competence. Further studies indicated that the apparent reductions in biosynthesis were not caused by decreased uptake of the labeled precursors by intact Wicky cells. In addition, these effects were probably not the result of turnover of macromolecules induced by CF. The lack of turnover of labeled peptidoglycan suggested that competence induction may not involve an autolysin.


Metabolic control of urea catabolism in Chlamydomonas reinhardi and Chlorella pyrenoidosa. In the unicellular green alga Chlamydomonas reinhardi (strain y-1), synthesis of the enzymes required for urea hydrolysis is under substrate induction control by urea and under end product repression control by ammonia. Hydrolysis of urea if effected by the sequential action of the discrete enzymes urea carboxylase and allophanate lyase, collectively called urea amidolyase. The carboxylase converts urea to allophanate in a reaction requiring biotin, adenosine 5'-triphosphate, and Mg2+. The lyase hydrolzyes allophanate to ammonium ions and bicarbonate. Neither activity is present in more than trace amounts when cultures are grown with ammonia or urea plus ammonia, or when they are starved for nitrogen for 8 h. Urea in the absence of ammonia induces both activities 10 to 100 times the basal levels. Addition of ammonia to an induced culture causes complete cessation of carboxylase accumulation and an 80% depression of lyase accumulation. Ammonia does not reduce urea uptake by repressed cells, so it does not prevent induction by the mechanism of inducer exclusion. The unicellular green alga Chlorella pyrenoidosa (strain 3 Emerson) also has discrete carboxylase and lyase enzymes, but only the carboxylase exhibits metabolic control.


Capacity of the outer membrane of a gram-negative marine bacterium in the presence of cations to prevent lysis by Triton X-100. Cells of marine pseudomonad B-16 (ATCC 19855) washed with a solution containing 0.3 M NaCl, 50 mM MgCl2, and 10 mM KCl (complete salts) could be protected from lysis in a hypotonic environment if the suspending medium contained either 20 mM Mg2+, 40 mM Na+, or 300 mM K+. When the outer double-track layer (the outer membrane) of the cell envelope was removed to yield mureinoplasts, the Mg2+, Na+ or K+, requirements to prevent lysis were raised to 80, 210, and 400 mM, respectively. In the presence of 0.1% Triton X-100, 220, 320, and 360 mM Mg2+, Na+ or K+, respectively, prevented lysis of the normal cells. Mureinoplasts and protoplasts, however, lysed instantly in the presence of the detergent at all concentrations of Mg2+, Na+, or K+ tested up to 1.2 M. Thus, the structure of the outer membrane appears to be maintained by appropriate concentrations of Mg2+ or Na+ in a form preventing the penetration of Triton X-100 and thereby protecting the cytoplasmic membrane from dissolution by the detergent. K+ was effective in this capacity with cells washed with complete salts solution but not with cells washed with a solution of NaCl, suggesting that bound Mg2+ was required in the cell wall membrane for K+ to be effective in preventing lysis by the detergent. At high concentrations (1 M) K+ and Mg2+, but not Na+, appeared to destabilize the structure of the outer membrane in the presence of Triton X-100.


Regulation of pyruvate dehydrogenase in isolated rat liver mitochondria. Effects of octanoate, oxidation-reduction state, and adenosine triphosphate to adenosine diphosphate ratio. Factors which influence the distribution of pyruvate dehydrogenase between its active, unphosphorylated form (PDHa) and its inactive, phosphorylated form (PDHb) have been examined in isolated rat liver mitochondria. A rapid freezing method was developed for the extraction of pyruvate dehydrogenase from incubated mitochondria which prevented interconversions between PHDa and PDHb which normally occur when mitochondria are collected by centrifugal methods. The intramitochondrial ATP:ADP ration was varied over a 100-fold range by the addition of dinitrophenol, oligomycin, or both substances to mitochondria oxidizing 2-oxoglutarate. PDHa activity was found to be inversely proportional to the intramitochondrial ATP:ADP ratio but was not closely correlated with the extramitochondrial adenine nucleotide levels. When mitochondria were incubated in State 4 with succinate and rotenone, the addition of pyruvate increased PDHa activity more than 10-fold without appreciably altering the mitochondrial ATP:ADP ratio. These observations are most readily explained by the known inhibitory effects of pyruvate and ADP on PDHa kinase. PDHa activity could be maintained at a high level by incubating mitochondria in a condition resembling State 3 by the addition of succinate, glucose, and hexokinase. The further addition of octanoate reduced PDHa activity by 60% without appreciably altering the ATP:ADP ratio. Rotenone had a sililar effect. When added in the presence of octanoate, rotenone further decreased PDHa activity whereas 4-pentenoate led to an increase in activity. The effects of octanoate on PDHa activity were not seen when mitochondria were incubated in the presence of high levels of pyruvate, though pyruvate oxidation was till diminished by over 50%. The data suggest that octanoate addition favors the PDHa kinase reaction leading to inactivation of PDHa, and in addition causes the accumulation of NADH and acetyl-CoA which are recognized competitive inhibitors of pyruvate dehydrogenase.