Impact of nutritional rehabilitation on enzymatic antioxidant levels in protein energy malnutrition.

To assess the role of enzymatic antioxidants in the pathogenesis of protein energy malnutrition (PEM) and the effect of nutritional rehabilitation, we studied 30 infants with PEM (mean age 10.63 +/- 4.39 months: 10 marasmic; 8 with kwashiorkor; 12 with marasmic kwashiorkor) and 15 controls. All underwent clinical examination and laboratory investigations, including superoxide dismutase (SOD) and glutathione peroxidase (GPx) estimation before and after nutrition rehabilitation. SOD and GPx were significantly lower in all malnourished infants compared to controls, and significantly increased after nutritional rehabilitation. These significant correlations suggest that antioxidants could be introduced during PEM nutritional rehabilitation to decrease morbidity and mortality.

Lysosomal enzymuria in protein energy malnutrition.

Protein energy malnutrition (PEM) is common in underprivileged populations in many parts of the world and results from diets deficient in protein (kwashiorkor) or protein and calories (marasmus). The literature documents renal tubular abnormalities in children with PEM. In PEM the reabsorption of amino acids and phosphate is defective. In many kidney disorders in which renal tubular function is impaired (e.g., diabetes, preeclampsia, nephrotic syndrome, sickle cell anemia), lysosomal enzymuria ensues. We compared the urinary excretion of the following five lysosomal enzymes in 31 Nigerian children with marasmus, kwashiorkor, or marasmic-kwashiorkor: beta-hexosaminidase, alpha-galactosidase, beta-galactosidase, beta-glucuronidase, and alpha-mannosidase. All of the protein energy malnourished children and the 18 age- and gender-matched controls were from the city of Jos, located in central Nigeria. In the severely malnourished children, the urine levels of all five lysosomal enzymes (expressed as units of enzyme activity per mg creatinine) were markedly increased. The greatest increases were seen with beta-hexosaminidase (16-fold) and beta-glucuronidase (14-fold). Routine clinical analyses also revealed that, relative to the control population, the sera of the 14 most severely malnourished patients contained 2- to 5-fold more vitamin B12 and markedly reduced levels (15%, p < 0.00001) of calcium. These data are significant in that they document lysosomal enzymuria in Nigerian children with severe PEM and point to the potential diagnostic utility of the urinary beta-galactosidase determination for assessing renal function in children with this disorder.

Glyoxalase I activity in erythrocytes from severely malnourished children

The enzyme glycoxalase I (glyox I) is involved in metabolic detoxification, and requires glutathione (GSH) as a cofactor. Given the low concentration of whole blood GSH in children with oedematous malnutrition, it is possible that the function of this pathway may be compromised in these children. Glyox I activity was therfore assayed in erythocytes taken from 133 severely malnourished children and 21 age-matched controls. The mean values (ñSEM) for the marasmic group (marasmus: 105 ñ 4/u/gm Hb) and the group with kwashiorkor (Kwash: 103 ñ 4/u/gm Hb) were not significantly different from controls (cont: 104 ñ 2u/gm HB)>. In the group with marasmic-kwashiorkor (M-K: 88 ñ 4u/g Hb) Glyox I activity was significantly lower in controls (p < 0.005), as well as in children with marasmus (p < 0.005), and kwashiorkor (p < 0.05). Enzyme activity was lower than normal in 45 percent of the MK group. Seven children died subsequent to admission; in five cases Glyox I activities were exceedingly low. There was a weak positive correlation between Glyox I activity and whole blood levels of GSH (r=0.215). We conclude that Glyox I activity is relatively unaffected in malnutrition, except in those with M-K and especially those who do not survive the acutely malnourished state (AU)

Glyoxalase I activity in erythrocytes from severely malnourished children

The enzyme glycoxalase I (glyox I) is involved in metabolic detoxification, and requires glutathione (GSH) as a cofactor. Given the low concentration of whole blood GSH in children with oedematous malnutrition, it is possible that the function of this pathway may be compromised in these children. Glyox I activity was therfore assayed in erythocytes taken from 133 severely malnourished children and 21 age-matched controls. The mean values (ñSEM) for the marasmic group (marasmus: 105 ñ 4/u/gm Hb) and the group with kwashiorkor (Kwash: 103 ñ 4/u/gm Hb) were not significantly different from controls (cont: 104 ñ 2u/gm HB)>. In the group with marasmic-kwashiorkor (M-K: 88 ñ 4u/g Hb) Glyox I activity was significantly lower in controls (p < 0.005), as well as in children with marasmus (p < 0.005), and kwashiorkor (p < 0.05). Enzyme activity was lower than normal in 45 per cent of the MK group. Seven children died subsequent to admission; in five cases Glyox I activities were exceedingly low. There was a weak positive correlation between Glyox I activity and whole blood levels of GSH (r=0.215). We conclude that Glyox I activity is relatively unaffected in malnutrition, except in those with M-K and especially those who do not survive the acutely malnourished state

Elevated glutathione S-transferase activity in erythrocytes from malnourished children.

Glutathione S-transferases (GSTs) are principally involved in detoxification. These enzymes can be induced by an increased flux of substrate, such as occurs during pro-oxidative stress or antioxidant deficiency. We tested the hypothesis that the postulated oxidative stress in severe malnutrition would result in induction of GSTs in erythrocytes. Erythrocyte GST activity towards 1-chloro-2,4-dinitrobenzene (CDNB) was measured in 271 malnourished children (22 undernourished; 92 marasmic; 82 kwashiorkor; 75 marasmic-kwashiorkor) and 48 healthy children. GST activity in the malnourished children was significantly higher than the control group (P < 0.01). The GST activity in the four classes of malnutrition did not differ. There was a weak relationship between GST activity and the height deficit, but not with the weight deficit, or the clinical features displayed by the children. The 11 children that died had a higher value than the survivors. There was no change in GST with anthropometric recovery. We conclude that erythrocyte GST has been induced in children with malnutrition. Induction of erythrocyte GST may be the result of exposure of the children to oxidative stress during the months prior to their presentation with severe malnutrition.

Elevated glutathione S-transferase activity in erythrocytes from malnourished children

Glutathione S-transferases (GSTs) are principally involved in detoxication. These enzymes can be induced by an increased flux of substrate, such as occurs during pro-oxidative stress or antioxidant deficiency. We tested the hypothesis that the postulated oxidative stress in severe malnutrition would result in induction of GSTs in erythocytes. Erythrocyte GST activity towards 1-chloro-2, 4-dinitrobenzene (CDNB) was measured in 271 malnourished children (22 undernourished; 92 marasmic; 82 kwashiorkor; 75 marasmic-kwashiorkor) and 48 healthy children. GST activity in the malnourished children was significnatly higher than the control group (p < 0.01). The GST activity in the four classes of malnutrition did not differ. There was a weak relationship between GST activity and the height deficit, but not with the weight deficit, or the clinical features displayed by the children. The 11 children that died had a higher value than the survivors. There was no change in GST with anthropometric recovery. We conclude that erythrocyte GST has been induced in children with malnutrition. Induction of erythrocyte GST may be the result of exposure of the children to oxidative stress during the months prior to their presentation with severe malnutrition (AU)

Red blood cell antioxidant enzyme concentrations in kwashiorkor and marasmus.

Kwashiorkor may occur when an imbalance between pro- and antioxidants in malnourished children results in an excess of free radicals. The concentrations of the antioxidant enzymes catalase (CAT), superoxide dismutase (SOD), reduced glutathione (GSH) and glutathione peroxidase (GPX) were measured in erythrocytes of 22 children with kwashiorkor on admission to hospital and repeated on days 5, 10 and 30 of recovery. The concentrations were compared with those in 22 children with marasmus and in 20 children who were normally nourished but had infective illness necessitating their hospitalization. CAT and SOD were similar in all groups and did not change during recovery. GSH and GPX were significantly lower in kwashiorkor than in the other groups. Concentrations of thiobarbituric acid-reactive substances (TBARS), a marker of lipid peroxidation, were significantly elevated in children with kwashiorkor. During clinical recovery, GSH but not GPX concentrations rose despite an increase in plasma selenium levels and decreased concentrations of TBARS. These findings suggest that the antioxidant status of children with kwashiorkor differs from that of well nourished and marasmic children. Whether these differences are the cause of the consequence of the clinical picture is unresolved.

Erythrocyte membrane (Ca2+ + Mg2+)-ATPase in human protein-energy malnutrition.

Calmodulin-free ghost membranes were prepared from erythrocytes of kwashiorkor children and from healthy children in the same age bracket. In the absence of calmodulin, the specific activity of Mg2+-dependent Ca2+-pumping ATPase (Ca2+ + Mg2+-ATPase) of kwashiorkor membranes was more than 40 percent lower than the specific activity of the normal enzymes, whose maximum velocity was increased by at least four-fold by the modulator protein. In contrast, the maximum velocity of the enzymes of kwashiorkor membranes was enhanced by calmodulin by about 1 1/2 times the basal activity of the normal enzymes and by 2 times the basal activity of the kwashiorkor enzymes. The affinity of the pump for ATP was lower in the membranes of kwashiorkor children (Km for ATP = 30.6 +/- 2.8 microM ATP) in comparison to normal membranes (Km for ATP = 21.7 +/- 2.0 microM ATP). Similarly, calmodulin-affinity of the enzymes, was lower in kwashiorkor membranes than in the normal membranes irrespective of source of calmodulin. Calmodulin from haemolysates of kwashiorkor red cells activated the enzymes of normal and kwashiorkor membranes to the same degree as calmodulin partially purified from the haemolysate of healthy children. A determination of the dependence of the activity of the pump on calcium in the absence and presence of calmodulin reveals that the affinity of the kwashiorkor enzymes for Ca2+ is at least 70 percent lower than that of enzymes of normal membranes. Altogether, these findings suggest that the Ca2+-pumping ATPase of kwashiorkor membranes is less functional than the enzymes of healthy erythrocytes.

Pepsins in protein-energy malnutrition.

This study reports on basal gastric pepsins in 40 normal, kwashiorkor and marasmic children admitted to the paediatric wards of the University of Benin Teaching Hospital, Nigeria. The activity of total pepsin was significantly depressed in the diseased states. The various fractions of pepsin separated on ion-exchange chromatography also showed dramatic reductions for both kwashiorkor and marasmus. This adaptive reduction of pepsin and its fractions was more drastic in marasmus than kwashiorkor. On disc-gel electrophoresis, four of the pepsin bands found in normal gastric aspirate were missing in children with the syndromes. Gastric electrolytes and acidity recorded for the diseased states were not conducive for maximal peptic activity. It is suggested that due to the reduced proteolytic capability of the malnourished infants, rehabilitation with intact protein should be cautious and gradual.

Serum aminotransferases and gamma-glutamyl transferase in protein energy malnutrition.

Forty six children suffering from Protein Energy Malnutrition (PEM) were classified according to the Wellcome classification. Their aspartate aminotransferase, alanine aminotransferase and gamma-glutamyl transferase were measured. Aspartate aminotransferase was raised in 20 patients (43.5%) and alanine aminotransferase was raised in 12 patients (26%). Y-glutamyl transferase was raised in only one patient suffering from marasmic kwashiorkor, who, in contrast to the rest of the patients had a marked rise in aminotransferases. The aminotransferase elevation correlated positively with a Severity Index calculated from height and weight retardation and serum albumin levels. It is suggested that the moderate rise in aminotransferases found in PEM is not due to damage to the liver. However, marked enzyme elevations can occur in a small minority of patients, suggestive of liver injury, probably caused by hepatotoxins.

Urinary excretion of arylsulfatases in malnourished/vitamin A deficient children.

Serum vitamin A (retinol) levels were generally low in all malnourished children (6-15 microgram/100 ml) compared with control children (50 microgram/100 ml). A significant increase in vitamin A after appropriate therapy was observed in all malnourished groups. Dietary supplements of proteins and calories even without extra vitamin A supplements increased serum vitamin A levels in cases of kwashiorkor indicating active mobilization of liver vitamin A. Total urinary arylsulfatase A activity excreted in 24-h or within 8-h in the morning (6 a.m. to 2 p.m.) was significantly reduced in cases of malnutrition with or without mild vitamin A deficiency symptoms. The excretion of arylsulfatase B was not altered. In cases of severe vitamin A deficiency coupled with malnutrition increased excretion of both arylsulfatases A and B was evident. These results on urinary arylsulfatases excretory pattern have been obtained either in samples collected for 24-h or specifically for 8-h (morning) and it is suggested that this test on urinary arylsulfatases may prove useful for detection of acute vitamin A deficiency with malnutrition in field studies. A ratio of arylsulfatases A/B of 2.0 or less seems to indicate mild malnutrition, the normal ratio being 3.4. Furthermore a low ratio coupled with increased excretion of both arylsulfatases A and B may be considered specific for acute vitamin A deficiency.

Infantile jejunal mucosa in infection and malnutrition.

Sixteen children with refractory diarrhea and three malnourished children who had frequent episodes of acute gastroenteritis but little diarrhea at the time of hospital admission, were studied by peroral upper small intestinal biopsy. Six children were adequately nourished; five children weighed 62 to 79% of expected weight and eight weighed less than 60% of expected weight. Two of the malnourished children had giardiasis. Pathogenic bacteria were found in only one case. Varying degrees of mucosal atrophy with reduction of mean villous height were seen in 18 cases. The concentration of mononuclear inflammatory cells and plasma cells was about half that seen in well-nourished children with severe nongastrointestinal infections. The concentration of mononuclear cells in the lamina propria was about twice that seen in normal adults. The proportions of IgA-producing cells and cells that stained for secretory component were significantly reduced, as compared with normal adult control values. This reduction was most striking in children with malnutrition complicated by giardiasis. Enzyme histochemical studies were performed for leucine aminopeptidase, alkaline phosphatase and acid phosphatase. There was a tendency for considerably reduced acid phosphatase activity in all clinical groups (kwashiorkor, marasmic kwashiorkor and marasmus) of growth-retarded infants.

Studies on hepatic and extrahepatic lipoprotein lipases in protein-calorie malnutrition.

Postheparin serum lipolytic activities (hepatic and extrahepatic), serum free fatty acid, and triglyceride levels were measured in 16 kwashiorkor, 14 marasmic, and 14 control children. The results showed that the reduction in total postheparin lipolytic activity in kwashiorkor was in the activity of hepatic origin. In marasmus, the total postheparin lipolytic activity, hepatic and extrahepatic activities, were within normal range. The was no evidence for the presence of inhibitors of postheparin lipolytic activity in the serum of kwashiorkor or marasmic children. Fasting serum-free fatty acid level was significantly elevated in kwashiorkor, while the level in marasmus was not significantly different from control value. The serum triglyceride levels in both conditions showed no significant differences from the control value. These findings suggest that the defective production of hepatic lipoprotein lipase, as well as increased influx of free fatty acid into the liver, could account for the accumulation of fat in the liver of kwashiorkor and not in that of marasmic children.

[Salivary amylase in kwashiorkor. III. Investigative study of salivary concentration and secretion in 4 patients presenting complications during nutritional recovery]. / Amilase salivar no kwashiorkor. III. Estudo evolutivo da concentração em quatro pacientes apresentando complicações durante a recuperação nutricional.

In four kwashiorkor children with ages between 20 and 36 months, the salivary volume (ml), the amylase concentration (U/ml) and secretion (U/Kg/5 min.) was studied, since the first day in the hospital until the discharge. These children had same diseases during the nutritional recuperation. The control group had eighteen children of the same age and eutrophic. The salivary secretion during 5 minutes was taken by special technic and in the same moment in each day. In one patient with serious kwashiorkor the level of the salivary secretion was slow while the nutritional state was bad. The levels of the salivary amylase secretion and concentration was slow while the disease was present making difficult the nutritional recuperation in each patient.

High erythrocyte membrane (Na+ + K+)-ATPase in kwashiorkor, in vivo reversal by diuretic.

Specific activity (mumol Pi released/h/mg membrane protein) of ouabain-sensitive (Na+ + K+)-ATPase has been shown to be higher in erythrocytes from children suffering from kwashiorkor, compared to that in normal children. Twenty four hours after treatment of these children with a diuretic, there was reduction in their body weights due to loss of oedema fluid. Ouabain sensitive (Na+ + K+)-ATPase of the erythrocyte membrane was inhibited by about 40% and this was associated with gain of 1.8 mequivalents Na+ per litre of erythrocytes. The results suggest that high ouabain-sensitive (Na+ + K+)-ATPase could be one of the mechanisms operative in erythrocytes to prevent accumulation of Na+ in kwashiorkor.

Erythrocyte membrane Na+ and K+ activated adenosine triphosphatase in protein-calorie malnutrition.

Total, ouabain insensitive and ouabain sensitive Na+ and K+ adenosine triphosphatase activity in the erythrocyte membrane of protein-calorie malnourished children with marasmus and kwashiorkor was compared with the enzyme activity in apparently healthy children (normals). Na+ and K+ contents of erythrocyte and plasma were also determined in these patients. Specific activity (units per milligram of membrane protein) of ouabain sensitive Na+ and K+ adenosine triphosphatase was significantly higher in erythrocyte membrane preparations from children with kwashiorkor but not from children with marasmus. After 4 to 5 weeks of treatment with diets sufficient in protein and calories the specific activity of the enzyme was lower as compared to that on admission. Erythrocyte and plasma Na+ content (microgram/10(6) cells and microgram per milliliter of plasma) in children with kwashiorkor were not different from those in normal children, however, there was reduction in K+ content of erythrocytes and plasma of these children. After treatment, erythrocyte Na+ and K+ and plasma K+ in children with kwashiorkor increased significantly. In marasmic children erythrocyte Na+ was higher as compared to normal but there were no differences in K+ content of either eyrthrocytes or plasma in these children.