Histidine utilization by Alcaligenes eutrophus: Regulation of histidase formation under heterotrophic conditions of growth

Histidine supported good growth of Alcaligenes eutrophus strain H 16 as a nitrogen source, but only poor growth as a carbon and energy source. The facultative chemolithoautotrophic bacterium was also able to utilize urocanic acid, the first intermediate of histidine catabolism. The products of histidine degradation were ammonium, formate and glutamate. Three enzymes of the pathway, histidase, urocanase and formiminoglutamate hydrolase, were present in histidine-grown cells. Two types of spontaneous mutants, derived from the wild type, were characterized by an increased growth rate on histidine. One of these types was found to produce histidase constitutively and at a higher activity compared with the parental strain. The second type of mutant had apparently gained an improved histidine uptake system, which is supposed to be growth rate-limiting in the wild type. From the physiological studies the conclusion was drawn that the control of histidine-degrading enzymes is based on induction by urocanate and catabolite repression by carbon sources supporting fast growth, such as succinate or pyruvate. Ammonium was found not to affect catabolite repression, however, we obtained evidence that histidine uptake is subject to a nitrogen control.Abbreviation CTAB hexadecyltrimethylammonium bromide     

Time course of active Na transport and oxidative metabolism following transepithelial potential perturbation in toad urinary bladder

Summary The use of an Ussing chamber with well-defined mixing characteristics coupled to a mass spectrometer permits the concurrent evaluation of transepithelial current and oxidative metabolism with improved temporal resolution. The time-course of the amiloride-sensitive currentI ^a and the rate of suprabasal CO₂ productionJ _CO2 ^sb were observed in 10 toad urinary bladders at short-circuit and after clamping at 100 mV, serosa positive. Following perturbation of (0100mV),I ^a declined sharply within 1/2 min, remained near constant 15 min, and then increased slightly.J _CO2 ^sb declined more gradually, remained near constant at 4–7 min, and then declined further. Detailed analysis revealed an early quasi-steady state with near constancy ofJ _CO2 ^sb starting at 2.9±1.1 (sd) min and lasting 4.7 ±1.8 (sd) min, followed by relaxation to a later steady state at about 15 min. During the early quasi-steady state,I ^a was also nearly constant. Considering that in steady statesI ^a/FJ _Na ^a , the rate of transepithelial active Na transport, during the early quasi-steady state mean values ±se ofJ _Na ^a ,J _CO2 ^sb and (J _Na ^a /J _CO2 ^sb ) were, respectively, 29.9±1.7%, 59.4 ±3.2%, and 56.4±5.7% of values at short-circuit. Corresponding values during the late steady state were 41.4±6.0%, 38.2±6.1%, and 111.3±8.6%. Thus the flow ratioJ _Na ^a /J _CO2 ^sb was depressed significantly during the early quasi-steady state, but returned later to the original value. The results of measurements ofI ^a andJ _CO2 ^sb in three hemibladders were qualitatively similar. In terms of a phenomenological black-box treatment the findings are consistent with earlier studies indicating incomplete coupling between transport and metabolism. Further studies will be required to clarify the molecular basis for these observations.     

Membranes of the protoplast L-form of Proteus mirabilis

Isolated membranes of the cell wall-less stable protoplast L-form of Proteus mirabilis were characterized by density gradient centrifugation and by assay for their major chemical constituents, proteins, phospholipids and lipopolysaccharide, and for some specific marker enzymes of the cytoplasmic membrane. In most of the analyzed properties the L-form protoplast membrane resembled the bacterial cytoplasmic membrane, with some notable modifications. considerable amounts of lipopolysaccharide, normally an exclusive constituent of the outer membrane, were found. Furthermore, the L-form membranes contained the functions of the reduced nicotinamide adenine dinucleotide oxidase system, of d-lactate dehydrogenase (EC 1.1.1.28) and of succinate dehydrogenase (EC 1.3.99.1) at specific activities comparable to, or in some cases considerably higher than, those present in cytoplasmic membranes of the bacterial form. Of two peptidoglycan DD-carboxypetidase/transpeptidases (EC 3.4.17.8 and EC 2.3.2.10), which are normally present in the cytoplasmic membrane of the bacterial form of P. mirabilis, the membrane of the protoplast L-form contained only one. Electron microscopy of thin sectioned L-form protoplasts showed extensive heterogeneity of membraneous structures. In addition to the single membraneous integument, internal membrane-bounded vesicles and multiple stacks of membranes were present, as the result of unbalanced growth and membrane synthesis in the L-form state.     

Physiological and biochemical contributions to the taxonomy of the genera Ankistrodesmus and Scenedesmus

The 17 Ankistrodesmus strains studied do not produce extracellular amylolytic enzymes. 20 out of 28 Scenedesmus strains, however, are able to hydrolyse starch.The groups of strains previously characterized physiologically and biochemically were assigned to established species of the genera Ankistrodesmus (including Monoraphidium) and Scenedesmus.Abbreviation GC Guanine + cytosine     

Two relative efficiencies of polymorphic enzymes for characterizing cell lines,detecting contaminations,and monitoring transplants

Summary A new calculation of the relative efficiency of polymorphic enzyme markers, called the REB, was determined and compared with one of Fisher's determinations of the relative efficiency called REA here. The REA estimates the chance of failing, and 1-REA of succeeding, to show a phenotypic difference between two randomly selected persons or cultured cell lines (Case 1). In this study it was shown that the REA also estimates the chance of detecting a cell line mislabeling or similar mixup (Case 2) and a cell line cross-contamination leading to the complete replacement of an original line by contaminating line (Case 3). The new REB determines the probability of failing, and 1-REB of succeeding, to detect a contamination of an original line by another line leading to their coexistence, or at least a sufficiently long period of transitional coexistence before one overgrows the other. The REA and REB also apply to determining the efficiency of polymorphic markers in detecting donor and recipient cells in tissue transplants. This work was developed from the author's involvement in the human tumor cell-line characterization project at Sloan-Kettering Institute and he acknowledges this opportunity and the benefits of his association with Dr. J?rgen Fogh and colleagues in the Human Tumor Cell Laboratory.     

Evidence for two genetic complementation groups in pyruvate carboxylase-deficient human fibroblast cell lines

We have examined genetic complementation in pyruvate carboxylase deficiency by comparing the enzyme activity in polyethylene glycol-induced heterokaryons with that in unfused mixtures of fibroblasts from three affected children. Complementation, manifested as a three- to sevenfold increase in pyruvate carboxylase activity, was observed in fusions between a biotin-responsive multiple carboxylase (pyruvate carboxylase, propionyl CoA carboxylase, and -methylcrotonyl CoA carboxylase) deficient fibroblast line and two other lines deficient only in pyruvate carboxylase activity. Kinetic analysis of complementing pyruvate carboxylase deficient lines, measured by the rate of restoration of enzyme activity as a function of time, revealed that maximum restoration was achieved within 10–24 hr after fusion. This profile is similar to those observed for fusions between the multiple carboxylase deficient line and two lines deficient in propionyl CoA carboxylase activity that are known to represent different gene mutations. Although the patients with pyruvate carboxylase deficiency had similar clinical findings, our studies indicate that pyruvate carboxylase deficiency is genetically heterogeneous, with at least two distinct, probably intergenic, complementation groups.This work was supported by an NIH research grant (AM 25675) and an A. D. Williams research grant (6-48360). B. Wolf is the recipient of an NIH Research Career Development Award (AM 00677) and is aided by a Basil O'Connor Starter Research Grant from The National Foundation-March of Dimes (5-263). G. Feldman is the recipient of an NIH predoctoral training grant (GM 07492). This article is No. 100 from the Department of Human Genetics at the Medical College of Virginia.     

Comparative sensitivities of electrophoretic assays for human enzymes

The sensitivities of 26 starch gel electrophoretic enzyme assays have been compared using HeLa human cells and A9 mouse cells grown in vitro.This research was supported by National Institutes of Health Grant No. USPHS GM 09966.     

Evolution des activités protéolytiques digestives chez les Jeunes Mulgilidae

Proteolytic activity in the stomach, pyloric caeca and intestine decreases with increasing size in the three fish species Mugil auratus, M. capito and M. saliens. Differences between these species are found mainly in the gastric proteolytic activity which appears to be related to diet. This activity is pronounced in M. saliens whose diet is mainly carnivorous. Comparison of regression lines relating gastric proteolytic activity to size reveals differences which distinguish M. auratus from both of the other species. Total proteolytic activity exhibits high variability depending on the types of diet.     

Isolubimin: A possible precursor of lubimin in infected potato slices

Potato slices treated with spirovetiva-1(10),11-diene-2-one yield lubimin and rishitin within 24 hr. A vetispirane which has not been detected in fungally infected potatoes was also isolated. This compound, isolubimin, appears to be an intermediate in the conversion of the above spirovetivone to lubimin.     

Local cold exposure of Andean Indians during normal and simulated activities.

Thermal resonses of Andean Indians were measured during several customary tasks associated with cold exposure in the highlands of southern Peru. These included surface temperature measurements of women while they washed clothing in the river and similar measurements of men while they constructed a diversion channel in the same river. A third test measured the effects of alcohol consumption on body temperatures during light activity. Women maintained slightly warmer hand than foot temperatures. Men maintained nearly equal hand and foot temperatures during the exposure period. Among male subjects the foot rewarmed at a faster rate than the hand. The results from the field studies compared favorably with results from earlier laboratory exposure tests. Comparisons between the river water exposure tests for males and females showed a consistent pattern where females maintained warmer hand and foot temperatures than males. These findings were in accord with previous laboratory studies among Quechua Indians and with the findings reported for other ethnic groups who experience natural cold stress. Alcohol ingestion appeared to have minimal effect in mitigating cold stress response during light activity. This finding was counter to earlier laboratory tests of resting subjects.