A sensitive method for determining quinolinic acid in animal tissues.
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Biomedical subjects
Publications and source records attributed to A Gil.
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DNA sequencing studies indicate that only one of two closely linked human embryonic alpha-like globin genes, zeta (zeta), encodes a functional polypeptide. The other is a pseudogene (psi zeta) that differs by only 3 bp in the protein coding sequence, one of which converts the codon for amino acid 6 into a chain termination codon. Both zeta-globin genes differ from all other alpha-like genes thus far reported in that they contain large introns consisting, in part, of simple repeat sequences. Intron 1 of each gene contains a variation of the repeat sequence ACAGTGGGGAGGGG, while intron 2 contains the repeat sequence CGGGG. Comparison of the human zeta- and alpha-globin gene sequences reveals that the embryonic and adult alpha-like genes began to diverge from each other relatively early in vertebrate evolution (400 million years ago). In contrast, the beta-like embryonic globin gene, epsilon (epsilon), is the product of a much more recent evolutionary event (200 million years ago). Thus, even though the temporal and quantitative expression of zeta- and epsilon-globin genes must be coordinately controlled during development, their evolutionary histories are clearly distinct.
The acid-soluble ribonucleotides of human milk were measured by enzymic and ion-exchange chromatographic procedures at different stages of lactation. Human colostrum and milk contained CMP, AMP, GMP, UMP, GDP-mannose, UDP-N-acetyl-glucosamine, UDP-N-acetyl-galactosamine, UDP-glucose, UDP-galactose, UDP and other minor nucleotides. Cytidine and adenosine derivatives were present in relatively higher amounts than in milk from ruminant species. The nucleotide concentration decreased with advancing lactation. Human milk at 3 months of lactation contained about 10 mumol/100 ml nucleotides, which represented about 75% of the amount of nucleotides present in human colostrum. Human milk did not contain orotic acid. The UDP-glucose/UDP-galactose ratio was constant during lactation and was similar to that of milks from ruminant species.
The evolution of plasma and urine amino acids in "small for dates" newborns from the first hours of life to 30 days has been studied. The results obtained point out the occurrence of malnutrition in these newborns who have lower levels of plasma albumin than controls, high indexes of malnutrition from plasma aminograms and lower urinary excretion of histidine and its N-methyl derivatives. All indexes are normalized at 7 days of life with the used diet.
The levels of quinolinic acid in liver and kidney and the gluconeogenic capacity in these tissues were determined in rats treated with tryptophan. The administration of this aminoacid produced a high increase in the hepatic quinolinic acid concentrations fed and 48 h. starved rats. On the contrary, only slight variations in the concentrations of renal quinolinic acid were observed. The highest value raised in these conditions was three times lower than Ki of the quinolinic acid for the phosphoenolpyruvate carboxykinase. These results suggest that tryptophan administration originates a selective inhibition of hepatic gluconeogenesis.
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Out of 110 cases of fever of unknown origin (FUO) that met Petersdorf and Beeson's criteria 15 patients were selected because of prolonged FUO with more than six months elapsed between admission and the final diagnosis. In this group of chronic FUO an etiological diagnosis was reached in 11 cases, distributed as follows: four cases with infections (two with toxoplasmosis, one with brucellosis, and another with a brain abscess); one with colon carcinoma; two with collagen-vascular diseases (systemic lupus erythematosus, temporal arteritis); and four with different diseases (two with familial mediterranean fever, one with idiopathic granulomatous disease, and another with factitious fever). In four cases no cause for the FUO could be determined. The procedures used to obtain the diagnosis were non-invasive in five cases (clinical course and serological tests), and invasive in another five (angiography, biopsies, and exploratory laparotomy). In one case the ethology could only be ascertained at autopsy. In the FUO with a prolonged course the peculiar etiological spectrum, the lesser yield of invasive procedures, and a mortality inferior to that of FUO in general all deserve special emphasis.
The appearance of amyloidosis during the course of multiple myeloma is a well known fact and has an overall incidence of 6 to 15%. However, the total transformation of a plasmocytoma into a voluminous amyloid tumor is a very rare event. A female patient was diagnosed of lambda light chain disease after developing a conspicuous rib plasmocytoma over the same region where a pathological fracture had appeared three years before. She was treated with discontinuous courses of melphalan and methyl-prednisolone, and developed a reversible nephrotic syndrome and a pathological fracture of the right clavicle. At necropsy there was generalized amyloidosis and complete substitution of the rib plasmocytoma by amyloid substance, with another important accumulation of amyloid in the region of the clavicular fracture. The present concepts on amyloidogenesis in multiple myeloma are reviewed, and the peculiarities of the present case together with the possible role of initiating factors and the effects of therapy are discussed. The case herein reported appears to represent a human model of focal amyloidogenesis in myeloma.
Renal function and renal size have been studied in ten early insulin-dependent diabetic patients and in ten matched control subjects. Glomerular filtration rate, renal plasma flow and radiological kidney size were determined in each subject. Glomerular filtration rate and renal plasma flow were increased in diabetics (mean +/- SD: 169.6 +/- 16.1 and 690.1 +/- 52.6 ml/min/1.73 m2, respectively) compared with controls (120.6 +/- 9.7 and 605.9 +/- 67.2 ml/min/1.73 m2; p less than 0.001 and p less than 0.01). Calculated kidney weight corrected to 1.73 m2 of body surface area was elevated in diabetics (385.2 +/- 29.0 g) with respect to controls (277.5 +/- 17.5 g; p less than 0.001). No significant differences were found between diabetics and control subjects when glomerular filtration rate was expressed per gram calculated kidney weight, while renal plasma flow was significantly lower in diabetics than control subjects when so expressed (p less than 0.01). A positive correlation was found between glomerular filtration rate, renal plasma flow and kidney size in both controls and diabetics (p less than 0.01 in all cases). These findings support the conclusion that in the early state of diabetes glomerular hyperfunction is related to enlargement of the kidneys and augmented renal plasma flow.
The acid-soluble ribonucleotides of cow's, goat's and sheep's milks were measured by enzymic and ion-exchange chromatographic procedures at different stages of lactation. Colostra and milk from ruminant species contained orotic acid and 13 well-identified nucleotides: AMP, CMP, GMP, UMP, UDP, GDP, UDP-glucose, UDP-galactose, UDP-N-acetyl-glucosamine, UDP-N-acetyl-galactosamine, UDP-glucuronate, GDP-mannose and GDP=fucose. Cow's goat's and sheep's colostrum contained significant amounts of nucleotides, which increased in amount from the moment of parturition, to reach a maximum 24--48 h later. The nucleotide concentration decreased thereafter with advancing lactation. Cow's milk contained substantial amounts of orotic acid, which increased during lactation, whereas in goat's and sheep's milk no increase took place. The cytidine and adenosine derivatives in ruminants' milk did not change significantly during lactation. The UDP-glucose/UDP-galactose ratio was measured in ruminants' milk at different stages of lactation.
Renal elimination of uric acid, calcium, phosphorus, sodium, potassium, chloride and magnesium and urinary acidification capacity were determined in ten insulin-dependent diabetics and in ten matched control subjects. The diabetics showed excessive excretion of uric acid, sodium, potassium, chloride and ammonia. Sodium, chloride and ammonia excretion fractions was also increased with respect to controls. The enhanced excretion of these substances in diabetics failed to relate to glomerular filtration rate, glycosuria or insulin requirements. These findings might be explained on the basis of glomerular filtration rate elevation, tubular response to this increment, and the underlying metabolic disturbances of diabetes.
Angiographic studies were carried out on 21 patients with systemic necrotizing vasculitis. Four basic arterial anomalies were found: (1) saccular microaneurysms appeared in 62% of the patients (2) arterial thrombosis was seen in 81% of patients; (3) arterial stenosis occurred in 81%; and (4) lumen irregularities occurred in 90%. Alterations in the renal vascular flow were also observed in accordance with changes in the cortical medullary differentiation, heterogeneous nephrogram, and prolonged washout. Two patients showed regression of microaneurysms after immunosuppressive therapy. We found angiography to be a low-risk technique of use for diagnosis and followup studies on vasculitis.
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Clinical symptoms, biochemical analyses, immunologic status and angiographic findings corresponding to seven HBsAg positive patients with panarteritis nodosa and to 16 HBsAg negative patients with panarteritis nodosa have been compared. HBsAg positive cases showed a statistical significant higher incidences of Raynaud's phenomenon (p less than 0.05) and cardiopathies (p les than 0.05), as well as high occurrence of blood hypertension, artropathy, liver involvement and peripheral neuropathy. Significant differences in relation to cell immunity were not found. Humoral immune disturbances were more common among HBsAg positive patients, being hypergammaglobulinemia (p less than 0.01), IgM increase and decrease of complement factors (C3, C4 and C3PA) the most frequent abnormalities recorded. Angiographic studies revealed a high incidence of microaneurisms for the HBsAg positive group.
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Gel filtration on Sephadex G-100 at pH 9.0 in 1 mM Tris buffer produces denaturation and inactivation of pancreatic DNAase A. Limiting concentrations of Ca2+ in the suspension and elution buffer, reactivates some of the enzyme molecules in an amount proportional to the calcium added. Stable active and inactive forms were separated on Sephadex columns. A model for the conformational role of Ca2+ on DNAase A demonstrates that at least one Ca2+ is involved (Kapp = 8.3 . 10(-5) M) in the correct folding of the polypeptide chain. Na+ was unable to reactivate the enzyme.
DNase A studied by gel filtration on Sephadex G-100 at pH 7.4 in 40 mM Tris-HCl buffer, behaves hydrodynamically as a spherical monomeric macromolecule of around 31,000 molecular weight, with a Stokes radius = 24.7 A, f/fo = 1.19, and D20,W = 8.69. Similar results were obtained by analytical dialysis using zinc chloride-modified cellophane membranes. The elution volume of DNase A decreases as the pH increases between pH 4.7 and pH 9.5. This effect has been attributed to a change in the tridimensional structure of the protein and interpreted as a modification in the axial ratio due to unfolding of the polypeptide chain with increase in the apparent Stokes radius. The addition of Ca2+ produce reversion of the pH-induced changes at pH 9.5. The transition occurs when Ca2+ binds to at least two binding sites (n = 1.66 in a Hill plot) with a Kd = 8.9 X 10(-5) M and the effect appears to be cooperative. These findings support the hypothesis that Ca2+-binding to DNase A causes a conformational change that maintains a more active structure of the enzyme, especially when the pH-induced unfolding reduces its activity.