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Biomedical subjects

M Losada

Publications and source records attributed to M Losada.

At least 55 records · Page 3Linked to original sources

Sustained Photoproduction of Ammonia from Dinitrogen and Water by the Nitrogen-Fixing Cyanobacterium Anabaena sp. Strain ATCC 33047.

Conditions have been developed that lengthen the time during which photosynthetic dinitrogen fixation by filaments of the cyanobacterium Anabaena sp. strain ATCC 33047 proceeds freely, whereas the subsequent conversion of ammonia into organic nitrogen remains blocked, with the resulting ammonia released to the outer medium. When l-methionine-dl-sulfoximine was added every 20 h, maximal rates of ammonia production (25 to 30 mumol/mg of chlorophyll per h) were maintained for about 50 h. After this time, ammonia production ceased due to a deficiency of glutamine and other nitrogenous compounds in the filaments, conditions which finally led to cell lysis. The effective ammonia production period could be further extended to about 7 days by adding a small amount of glutamine at the end of a 40-h production period or by allowing the cells to recover for 8 h in the absence of l-methionine-dl-sulfoximine after every 40-h period in the presence of the inhibitor. A more prolonged steady production of ammonia, lasting for longer than 2 weeks, was achieved by alternating treatments with the glutamine synthetase inhibitors l-methionine-dl-sulfoximine and phosphinothricin, provided that 8-h recovery periods in the absence of either compound were also alternated throughout. The biochemically manipulated cyanobacterial filaments thus represent a system that is relatively stable with time for the conversion of light energy into chemical energy, with the net generation of a valuable fuel and fertilizer through the photoreduction of dinitrogen to ammonia.

Journal Article↗

Purification and properties of glutathione reductase from the cyanobacterium Anabaena sp. strain 7119.

An NADPH-glutathione reductase (EC 1.6.4.2) has been purified 6,000-fold to electrophoretic homogeneity from the filamentous cyanobacterium Anabaena sp. strain 7119. The purified enzyme exhibits a specific activity of 249 U/mg and is characterized by being a dimeric flavin adenine dinucleotide-containing protein with a ratio of absorbance at 280 nm to absorbance at 462 nm of 5.8, a native molecular weight of 104,000, a Stokes radius of 4.13 nm, and a pI of 4.02. The enzyme activity is inhibited by sulfhydryl reagents and heavy-metal ions, especially in the presence of NADPH, with oxidized glutathione behaving as a protective agent. As is the case with the same enzyme from other sources, the kinetic data are consistent with a branched mechanism. Nevertheless, the cyanobacterial enzyme presents three distinctive features with respect to that isolated from non-photosynthetic organisms: (i) absolute specificity for NADPH, (ii) an alkaline optimum pH value of ca. 9.0, and (iii) strong acidic character of the protein, as estimated by column chromatofocusing. The kinetic parameters are very similar to those found for the chloroplast enzyme, but the molecular weight is lower, being comparable to that of non-photosynthetic microorganisms. A protective function, analogous to that assigned to the chloroplast enzyme, is suggested.

Amino Acids↗

Optimization of Conditions for Photoproduction of Ammonia from Nitrate by Anacystis nidulans.

The effect of several relevant environmental factors influencing the photoproduction of ammonia from nitrate by Anacystis nidulans cells treated with the glutamine synthetase inhibitor l-methionine-dl-sulfoximine has been investigated. The optimal ratio between l-methionine-dl-sulfoximine concentration (micro-molar) and cell density (micrograms of chlorophyll per milliliter) was around 1, the process taking place at maximal rate at a temperature of about 40 degrees C, within the pH range of 7 to 10. Ammonia production was stimulated by CO(2) or bicarbonate and was not affected by the accumulation of ammonia in the medium up to concentrations of 30 mM. The rate of ammonia production was found to be determined by the interaction of at least four factors, namely, irradiance and the density, depth, and turbulence of the cell suspension. Ammonia photoproduction from nitrate and water represents an interesting process for the conversion of light energy into chemical energy, which can operate at high efficiency, around 30% of its theoretical maximum.

Journal Article↗

Sustained Photoproduction of Ammonia from Nitrate by Anacystis nidulans.

Conditions that lengthen the time during which l-methionine-dl-sulfoximine (MSX) promotes excretion of ammonia produced by photosynthetic nitrate reduction in Anacystis nidulans have been sought. If MSX was added every 24 h, maximal rates of ammonia production were maintained for 3 days. After this time, ammonia production ceased due to a specific deficiency of glutamine in the cells, which finally led to cell lysis. The effective ammonia production period could be further extended either by adding a low amount of glutamine at the end of the 3-day period or by allowing the cells to recover for 8 h in the absence of MSX after every 48-h period in the presence of inhibitor. In this way, a steady production of ammonia lasting for at least 10 days was achieved. The MSX-treated cyanobacterial cells thus represent a system relatively stable with time for the conversion of light energy into chemical energy through the photoreduction of nitrate to ammonia.

Journal Article↗

DNA release and appearance of antinuclear antibodies in chronic hemodialysis patients.

The presence of free serum DNA and/or antibodies against nuclear antigens was studied in 65 patients in chronic hemodialysis. Most of patients showed increased levels of both free single-stranded DNA (ss DNA) (28 out of 29) and native (double-stranded) DNA (n DNA) (25 our of 28) at the beginning of each dialysis session (69 +/- 26 vs. 119 +/- 14, p less than 0.05) suggesting a rapid in vivo degradation of the n DNA released. 15 out of 65 patients (23%) developed low anti-ss DNA antibody titers by Millipore filtration assay. 2 of them presented anti-n DNA antibodies simultaneously. Isolated anti-n DNA, anti-Sm and anti-RNP antibodies were systematically negative in all patients. No relationship was found between the presence of anti-ss DNA antibodies and the type of nephropathy, the duration of treatment and the method of dialysis. The fate of a renal allograft was studied in 12 patients after having measured serum-free DNA and anti-DNA antibodies. The 4 patients with good graft function after 2 years of transplantation had significantly higher circulating ss-DNA levels than the other 8 patients whose grafts were rejected.

Antibodies, Antinuclear↗

Mesangial deposits of IgM in patients with the nephrotic syndrome.

Kidney biopsies from fourteen patients with the nephrotic syndrome were studied by light, immunofluorescence and electron microscopy. Morphologically, all cases showed moderate diffuse mesangial cell proliferation. In all cases, immunofluorescent microscopy demonstrated diffuse and generalized deposits of IgM as the sole or predominant immunoglobulin. Electron microscopic examination showed electron dense deposits localized in the mesangium in seven cases. One nephrotic patient experienced spontaneous remission and eight others were steroid responsive. Only one of the five steroid resistant patients treated with chlorambucil showed clinical remission. Five steroid responsive patients relapsed. At present, four patients are healthy, having not relapsed for the past two years. Although deposits of IgM may be related to the renal pathology in cases of the nephrotic syndrome, immunofluorescence findings do not seem to be sufficiently consistent or characteristic to justify a possible subclassification of the idiopathic nephrotic syndrome.

Adolescent↗

Human perinatal distribution of butorphanol.

The perinatal distribution of butorphanol was demonstrated in relation to maternal-neonatal transfer and colostrum/milk excretion in obstetric patients. Parenteral butorphanol passed the placental barrier and was found in neonatal cord serum. The mean neonatal serum concentration of butorphanol was not different from the mean maternal serum concentration of butorphanol, following a 1 or 2 mg intramuscular dose. Butorphanol was detected in the milk of lactating women following oral and intramuscular administration. Serum and milk concentrations appeared to be parallel with time. This observation was confirmed by the constancy of the mean milk-to-serum concentration ratio (0.7 intramuscular, 1.9 oral). We calculated that 4 micrograms would be the maximum amount of butorphanol, which would be expected to be present in the full daily milk output (1 L) following administration four times a day of 2 mg intramuscular or of 8 mg oral doses. An oral dose of 4 micrograms to an infant weighing 4 kg corresponds to the negligible oral dose of 0.7 mg to a 70 kg adult. The demonstrated safety and efficacy of butorphanol as an obstetric analgesic and the characteristics of the maternal-neonatal transfer and milk excretion are indicative of the potential excellence of this agent for obstetric use.

Anesthesia, Obstetrical↗

Purification and properties of assimilatory nitrate reductase [NAD(P)H] from Ankistrodesmus braunii.

Assimilatory nitrate reductase [NAD(P)H] (EC 1.6.6.2) from Ankistrodesmus braunii has been purified to homogeneity by a simple procedure that utilizes as the main step affinity chromatography on Blue-Sepharose. The best enzyme preparation has a specific activity of 61.25 units/mg protein. The enzyme has a sedimentation coefficient of 10.9 S by sucrose-density-gradient centrifugation, and a Stokes radius of 9.8 nm was estimated by gel filtration techniques. Its molecular weight is 460000, but only one single band of 58000 was detected after sodium dodecyl sulfate/polyacrylamide gel electrophoresis. The native enzyme seems thus to be composed of eight subunits. The nitrate reductase absorption spectrum shows wavelengths maxima at 280 and 416 nm and a broad shoulder at 450 nm. Reduced enzyme shows maxima at 424 (Soret), 527 (beta) and 557 (alpha) nm, and a bleaching at 450 nm. The reduced extracted heme chromophore, in pyridine and KOH, shows absorption bands at 414, 522 and 552 nm. These properties indicate the presence of a b-type cytochrome and flavin as prosthetic groups of A. braunii nitrate reductase. A minimum of four molecules of heme has been calculated per molecule of the enzyme complex. Redox titration of the enzyme shows a midpoint potential for the heme of -73 mV at pH 7.0. In the presence of p-hydroxymercuribenzoate, which inhibits the NAD(P)H-dependent activities of the complex, the enzyme-bound heme can be reduced with dithionite, but not with NAD(P)H.

Centrifugation, Density Gradient↗

Double-blind oral analgesic study of butorphanol in musculoskeletal pain: a comparison with codeine and placebo.

Butorphanol tartrate (4 mg and 8 mg) was compared to codeine phosphate (60 mg) and placebo for oral analgesic activity and side-effects employing a double-blind design in ninety-three out-patients suffering from moderate to very severe musculoskeletal pain. The study duration was 72 hours with medication administered every 4 to 6 hours (four times daily) for a total of twelve doses per patient. The results demonstrate that both the 4 mg and 8 mg doses of butorphanol were significantly better (p less than 0.u5) than placebo. While codeine 60 mg also proved active, it appears to be less efficacious than the high dose of butorphanol. The peak effect appeared to be evident in 1 to 2 hours. Butorphanol may be at least seven times more potent than codeine on a milligram basis. Although no serious side-effects were observed, butorphanol appeared to present a greater incidence of side-effects than codeine and placebo in this study.

Adolescent↗

Comparison of analgesia by intravenous butorphanol and meperidine in patients with post-operative pain.

Intravenous doses of butorphanol tartrate (0.5 mg, 1.0 mg and 2.0 mg) and meperidine hydrochloride (20 mg and 40 mg) were compared under controlled conditions employing a double blind study design. Informed consent was obtained from all post-operative patients suffering from moderate to severe pain who participated in this study. Approximately 25 patients were included in each group. The data from 125 patients were subjected to statistical analysis. The results indicated that butorphanol is approximately 40 to 50 times more potent than meperidine. In addition, at most of the time intervals, there were no statistically significant differences between the responses to butorphanol 0.5 mg and 1 mg and meperidine 20 mg and 40 mg; but the response to butorphanol 2 mg was significantly (p less than 0.05) better than the low dose of each agent. The low doses of butorphanol (0.5 mg) and meperidine (20 mg) appear to have an effective duration of action of less than two hours. The larger doses (butorphanol 1.0 mg and 2.0 mg and meperidine 40 mg) appeared to produce a two- to four-hour duration of action. The largest butorphanol dose (2.0 mg) appeared to produce the longest duration of action. A comparison of the test groups with respect to the incidence and type of side effects showed that butorphanol 2.0 mg produced a greater incidence of drowsiness (39 per cent). The overall incidence of drowsiness for patients receiving either the 0.5 mg or 1.0 mg dose of butorphanol was 12 per cent, as compared with an 8 per cent overall incidence in the meperidine group. The incidence of other side effects was relatively low in all test groups. No significant differences were noted among the groups with regard to the onset (usually less than or equal to 30 minutes post-therapy) or the duration (usually less than or equal to 2 hours) of side effects. Butorphanol appears to be a safe and effective analgesic for the relief of moderate to severe post-operative pain.

Adult↗