Spatial and temporal patterns of temperature,alkalinity, dissolved oxygen and conductivity in an oligo-mesotrophic,deep-storage reservoir in central Texas

Impoundment behavior was determined for alkalinity, temperature, dissolved oxygen, and conductivity from stations located along the length of a bottom-draining, oligo-mesotrophic, hardwater, deep-storage reservoir located in central Texas. The epilimnion deepened the length of the reservoir throughout the summer as a result of drawdown. Bicarbonate alkalinity and conductivity exhibited both longitudinal and vertical stratification. Alkalinity and conductivity in the epilimnion decreased from the riverine reach downreservoir to the dam. This longitudinal progression was attributed to inflow and photosynthetically induced epilimnetic decalcification.Hypolimnetic anoxic conditions first occurred in the sedimentation zone in the upreservoir and riverine reaches and then developed in a downreservoir pattern as summer progressed as a result of drawdown. Alkalinity and conductivity in the hypolimnion increased during anoxic conditions and consequently increased in a downreservoir progression.     

Bacterial breakdown of benomyl. II. Mixed cultures

Experiments with mixed bacterial cultures grown in liquid media which contained the benzimidazole fungicide benomyl, with or without Na-lactate, as source of carbon provided circumstantial evidence for cleavage of the benzimidazole heterocyclic ring. Yet, neither 2-aminobenzimidazole (2-AB) nor benzimidazole, as sole source of carbon, supported any bacterial growth. Total ¹⁴C-balance analysis experiments conclusively showed production of ¹⁴CO₂ from [2-¹⁴C] methyl benzimidazol-2-yl carbamate (MBC), and thus cleavage of the benzimidazole nucleus; bioassays, however, showed that the actual rate of benomyl and MBC breakdown was only small, the parent compound benomyl being still recovered in substantial quantities after up to 80 days of incubation. Therefore, cleavage of the benzimidazole ring is probably a matter of cometabolism, n-butylamine which originates from the butylcarbamoyl side chain serving as the proper source of carbon.Besides radiolabelled 2-AB and CO₂, an unknown metabolite was isolated which showed characteristics of a 2-AB-nucleotide. Probably, 2-AB was incorporated into bacterial DNA, which upon lysis of the bacterial cells gave rise to the nucleotide in question. Therefore, 2-AB might exert its inhibitory action by interfering with the normal functioning of DNA.     

On the relation between filament overlap and the number of calcium-binding sites on glycerinated muscle fibers.

The formation of rigor complexes between the thick and thin filaments of glycerinated rabbit psoas muscle fibers causes the fibers to bind more calcium at any given level of free calcium. I studied the maximum amount of calcium bound as a function of filament overlap under rigor conditions. Fibers stretched to zero filament overlap (sarcomere length greater than 3.8 micron) bound exactly 75% as much calcium as fibers with maximum overlap. Between these extremes a linear relationship was found between maximum bound calcium and the length of the overlap zone. The results support the hypothesis that in the intact filament lattice one of the four calcium-binding sites of troponin depends for its existence on attachment between myosin and actin. In addition, the linear relation between maximum bound calcium and filament overlap is consistent with the assumption that the cooperative effect of rigor complex formation on calcium binding is limited to the binding site in the immediate vicinity of the rigor complex.     

Further evidence for a phycobilisome model from selective dissociation,fluorescence emission,immunoprecipitation, and electron microscopy

Phycobilisomes, isolated in 500 mM Sorensen's phosphate buffer pH 6.8 from the red alga, Porphyridium cruentum, were analyzed by selective dissociation at various phosphate concentrations. The results are consistent with a structural model consisting of an allophycocyanin core, surrounded by a hemispherical layer of R-phycocyanin, with phycoerythrin being on the periphery. Such a structure also allows maximum energy transfer.Intact phycobilisomes transfer excitation energy ultimately to a pigment with a fluorescence emission maximum at 675 nm. This pigment is presumed to be allophycocyanin in an aggregated state. Uncoupling of energy transfer among the pigments, and physical release of the phycobiliproteins from the phycobilisome follow a parallel time-course; phycoerythrin is released first, followed by R-phycocyanin, and then allophycocyanin. In 55 mM phosphate buffer, the times at which 50% of each phycobiliprotein has dissociated are: phycoerythrin 40 min, R-phycocyanin 75 min, and allophycocyanin 140 min.The proposed arrangement of phycobiliproteins within phycobilisomes is also consistent with the results from precipitation reactions with monospecific antisera on intact and dissociated phycobilisomes. Anti-phycoerythrin reacts almost immediately with intact phycobilisomes, but reactivity with anti-R-phycocyanin and anti-allophycocyanin is considerably delayed, suggesting that the antigens are not accessible until a loosening of the phycobilisome structure occurs. Reaction with anti-allophycocyanin is very slow in P. cruentum phycobilisomes, but is much more rapid in phycobilisomes of Nostoc sp. which contains 6–8 times more allophycocyanin. It is proposed that allophycocyanin is partially exposed on the base of isolated intact phycobilisomes of both algae, but that in P. cruentum there are too few accessible sites to permit a rapid formation of a precipitate with anti-allophyocyanin.Phycobilisome dissociation is inversely proportional to phosphate concentration (500 mM to 2 mM), and is essentially unaffected by protein concentration in the range used (30–200 μg/ml). Phycobiliprotein release occurs in the same order (phycoerythrin > R-phycocyanin > allophycocyanin) in the pH range 5.4–8.0.     

Phosphatidic acid phosphatase and phospholipase A activities in plasma membranes from fusing muscle cells

Plasma membranes from fusing embryonic muscle cells were assayed for phospholipase A activity to determine if this enzyme plays a role in cell fusion. The membranes were assayed under a variety of conditions with phosphatidylcholine as the substrate and no phospholipase A activity was found. The plasma membranes did contain a phosphatidic acid phosphatase which was optimally active in the presence of Triton X-100 and glycerol. The enzyme activity was constant from pH 5.2 to 7.0, and did not require divalent cations. Over 97% of the phosphatidic acid phosphatase activity was in the particulate fraction. The subcellular distribution of the phosphatidic acid phosphatase was the same as the distibutions of the plasma membrane markers, $\text{Na}{}^{\mathrm{+}}\text{+ K}{}^{\mathrm{+}}$)-ATPase and the acetylcholine receptor, which indicates that this phosphatase is located exclusively in the plasma membranes. There was no detectable difference in the phosphatidic acid phosphatase activities of plasma membranes from fusing and non-fusing cells.     

Investigations on the turnover of adrenocortical mitochondria

Summary The effects of chronic administration of ACTH (up to 36 consecutive days) on the mitochondria of the zona reticularis of the rat adrenal cortex were investigated by stereologic techniques. It was found that ACTH induces two phases of hypertrophy of mitochondria alternating with two proliferative stages, which are associated with a significant decrease in the average volume of the organelles. It is suggested that, as in the zona fasciculata, ACTH controls the processes of growth and division of mitochondria in the zona reticularis. The mechanism underlying this action of ACTH as well as the differences between the responses to ACTH of the mitochondrial population of the two adrenal zones are discussed in the light of evidence indicating that mitochondria contain a complete genetic apparatus largely independent of nuclear control.The authors wish to thank Miss A. Coi and Mr. G. Gottardo for their excellent technical assistance. This work was partly supported by a contract with the CNR (C.T. 73.00663.04)     

In vitro anti-hapten response to a lactoside-conjugated protein

This paper describes the in vitro response of mouse lymphocytes to the hapten, azophenyl lactoside (Lac), coupled to the protein carrier, keyhole limpet hemocyanin, (KLH), and attempts to characterize and quantify the specific B and T cells involved. The system studied in detail has used spleen suspensions from mice which received Lac-KLH in complete adjuvant some months previously and a subsequent iv injection. The mean IgG plaque response of such cultures was about 15,000 with a corresponding IgM response about 5% of that value, similar in magnitude and Ig class distribution to in vivo responses. Primary responses were small, reflecting the low frequency of Lac precursors in normal mice. From dilution studies, we have estimated that the frequency of precursors specific for the Lac epitope in primed and normal mice is of the order of 10^?5 and 10^?7, respectively. The clonal yield of plaques from a stimulated B cell was about 150, independent of T cell concentration. The Lac response was dependent on the presence of adherent cells. It also shows a stringent requirement for carrier-specific T cells, which could not be satisfied by nonspecific T cell products or the addition of B cell mitogens. We have found that both the IgM and the IgG Lac responses are dependent on specific T cell help.     

The active species of 'CO2' utilized by reduced ferredoxin:CO2 oxidoreductase from Clostridium pasteurianum.

Reduced ferredoxin:CO2 oxidoreductase (CO2 reductase) from Clostridium pasteurianum catalyzes the reduction of 'CO2' to formate with reduced ferredoxin, an isotopic exchange between 'CO2' and formate in the absence of ferredoxin, and the oxidation of formate to 'CO2' with oxidized ferredoxin. The active species of 'CO2', i.e. CO2 or HCO3 (H2CO3), utilized by the enzyme was determined. The method employed for the species identification was that of Copper et al. (1968). Both 'CO2' reduction to formate and the exchange reaction were studied. Data were obtained which are compatible with those expected if CO2 is the active species. The V and the dissociation constant Ks of the enzyme - CO2 complex in dependence of pH were determined from initial velocity studies of the exchange reaction. V was found to be only slightly affected by pH between 5.5 and 7.5. Ks was markedly dependent on pH; the constant increased with decreasing pH from 0.2 mM at pH 7.5 to 3 mM at pH 5.5.