Heinz Body Counts Research Articles

PSXV-15 Effect of Supplemental Garlic on the Incidence of Heinz Body Anemia in non-Working Horses

Abstract Garlic (Allium sativum) has been historically used as a fly and pest control for horses, a practice that continues today. Recent research has shown garlic supplementation may cause Heinz body anemia in horses. The purpose of this study was to evaluate the incidence of Heinz body anemia in horses supplemented with incremental amounts of powdered garlic. The study included 12 horses divided into 4 groups (n=3), with horse serving as the experimental unit. Supplementation groups included: Control (0.0 g garlic/kg BW and 14.3 g molasses) and 3 supplementation rates: Low (0.0625 g garlic/kg BW and 14.3 g molasses), Medium (0.125 g garlic/kg BW and 14.3 g molasses), and High (0.1875 g garlic/kg BW and 14.3 g molasses) that were fed daily for 74 days. Horses were maintained on dormant tall fescue pasture and had ad libitum access to medium quality tall fescue hay throughout the study. Blood samples (jugular venipuncture) and weights were taken on day 0, 25, 50, and 74. Data were analyzed using Proc Mixed (SAS 9.4) for a completely randomized design, α = 0.05. Garlic supplementation reduced red blood cell count (P = 0.028) compared with Control and lymphocyte count (P = 0.046) showed a quadratic response to supplementation rate, with Low having the greatest lymphocyte count. A trend was noted for white blood cell count (P = 0.090) and hemoglobin count (P = 0.074) with garlic supplementation. Heinz body count (P = 0.406) was not affected by garlic supplementation. Results are interpreted to mean that garlic supplementation did not induce Heinz body anemia in horses; however, other blood constituents, including red blood cell and lymphocyte counts, were affected. Horses in the study were non-working, further research would be needed to evaluate any potential health impacts these variations in blood constituent concentrations would have in working horses.

Therapeutic Effect of Hydrogen Injected Subcutaneously on Onion Poisoned Dogs.

IntroductionThe purpose of this study was to investigate the therapeutic effect of hydrogen on the therapy of onion poisoned dogs.Material and MethodsA total of 16 adult beagle dogs were divided into two groups (control and hydrogen) and all were fed dehydrated onion powder at the dose of 10 g/kg for three days. The dogs of the experimental group were given subcutaneous injection of 0.2 mL/kg of hydrogen for 12 days after making the poisoned model successful. Blood samples were collected before feeding onions, one day before injecting hydrogen, and 2 h after the injection of hydrogen on days 1, 3, 5, 7, 9, and 12. Control dogs were not treated with hydrogen.ResultsThe levels of leukocyte production, anaemia, red blood cell degeneration which was reflected by the values of Heinz body count, haemolytic ratio, and oxidative products in hydrogen treated group were lower than in control dogs on some days. The capacity of medullary haematopoiesis that was based on reticulocyte counts, and the antioxidation in hydrogen group were higher compared with control group. However, the differences in renal function were not obvious in both groups.ConclusionAccordingly, it was concluded that subcutaneous injection of hydrogen could alleviate the symptoms in onion poisoned dogs.

Hematology of healthy Florida manatees (Trichechus manatus)

Hematologic analysis is an important tool in evaluating the general health status of free-ranging manatees and in the diagnosis and monitoring of rehabilitating animals. The purpose of this study was to evaluate diagnostically important hematologic analytes in healthy manatees (Trichechus manatus) and to assess variations with respect to location (free ranging vs captive), age class (small calves, large calves, subadults, and adults), and gender. Blood was collected from 55 free-ranging and 63 captive healthy manatees. Most analytes were measured using a CELL-DYN 3500R; automated reticulocytes were measured with an ADVIA 120. Standard manual methods were used for differential leukocyte counts, reticulocyte and Heinz body counts, and plasma protein and fibrinogen concentrations. Rouleaux, slight polychromasia, stomatocytosis, and low numbers of schistocytes and nucleated RBCs (NRBCs) were seen often in stained blood films. Manual reticulocyte counts were higher than automated reticulocyte counts. Heinz bodies were present in erythrocytes of most manatees. Compared with free-ranging manatees, captive animals had slightly lower MCV, MCH, and eosinophil counts and slightly higher heterophil and NRBC counts, and fibrinogen concentration. Total leukocyte, heterophil, and monocyte counts tended to be lower in adults than in younger animals. Small calves tended to have higher reticulocyte counts and NRBC counts than older animals. Hematologic findings were generally similar between captive and free-ranging manatees. Higher manual reticulocyte counts suggest the ADVIA detects only reticulocytes containing large amounts of RNA. Higher reticulocyte and NRBC counts in young calves probably reflect an increased rate of erythropoiesis compared with older animals.

An experimental study of hemolysis induced by onion (Allium cepa) poisoning in dogs

The mechanism of hemolysis induced by onion poisoning in dogs was studied. Six adult, clinically normal Pekingese dogs were fed cooked onions at 30 g/kg body weight/day for 2 days. Blood samples were collected on days 1, 3, 5, 8, 12, 18 and 24 after onion administration, and urine was collected the day after bleeding. Red blood cell counts, hemoglobin and hematocrit were decreased from day 1, and significantly so on day 5 (P < 0.01), contrary to the results of white blood cell counts. So the plasma bilirubin levels and urobilinogen were increased on day 3 (P < 0.01) and day 4 (P < 0.01), respectively. The Heinz body counts were increased dramatically from day 1 (P < 0.01), peaking on day 3 (P < 0.01). Reticulocyte counts were increased from day 1 and the highest value was on day 8 (P < 0.01). Besides anemia, the following erythrocyte parameters were altered: erythrocyte glucose-6-phosphate dehydrogenase was reduced from day 1 and reached the lowest value on day 5 (P < 0.01); the reduced form of nicotinamide adenine dinucleotide phosphate was decreased on day 1 (P < 0.01); reduced glutathione was decreased from day 1 (P < 0.01) and the lowest value was on day 3 (P < 0.01); glutathione-peroxidase was increased on day 1, but decreased significantly on day 3 (P < 0.01); hydrogen peroxide was increased on day 1 (P < 0.01), then went down on days 3-12 (the undermost value on day 5, P < 0.01); catalase was risen dramatically on day 5 (the peak value, P < 0.01); malondialdehyde (MDA) was increased on days 1-8, (P < 0.01), and reached the peak value on day 5 (P < 0.01). Deformity of the erythrocyte membrane was decreased on days 1-12 and fluorescence polarization (rho) and microviscosity (eta) of the erythrocyte membrane were increased on days 1-12 (P < 0.01). There were positive correlations between MDA and rho as well as eta, with correlation coefficients of 0.908 and 0.922, respectively (P < 0.01), but there was a negative correlation between MDA and deformity index, with a correlation coefficient of -0.887 (P < 0.05). This study confirmed that onion poisoning in dogs causes hemolytic anemia.

Efetividade da vitamina E na redução do estresse oxidativo em hansenianos da forma multibacilar sob tratamento

Abstract The disease, multibacillary leprosy, is treated with dapsone,clofazimine and rifampicin, with this therapy causing oxidativestress, leading to methemoglobinemia and the formation of Heinzbodies. Due to its antioxidant action, vitamin E has been used toprevent oxidative stress. Our objective was to evaluate theeffectiveness of vitamin E in patients with multibacillary leprosywith the hypothesis that this medication may reduce the oxidativestress produced. The study involved 32 patients with multibacillaryleprosy being treated with dapsone, clofazimine and rifampicin.The presence of oxidative stress was initially confirmed andsubsequently the patients were randomly divided into two equalgroups denominated as group “taking vitamin E” and “control”. Thepatients of the group “taking vitamin E” orally ingested 800 UI/dailyof vitamin E and the group “control” did not take any vitaminsupplement. After 30, 60 and 90 days of treatment, blood samples ofthe two groups were drawn to determine the metahemoglobinconcentration, presence of Heinz bodies and ferritin levels. The resultsof ferritin levels were submitted to the Mann-Whitney statistical testand the results of methemoglobin concentrations and Heinz bodycounts were submitted to the Fisher’s exact test. There were nosignificant differences between the two groups. In conclusion, vitaminE at the dose and for the treatment duration used, neither protectedagainst the oxidative stress caused by dapsone, clofazimine andrifampicin used by patients with multibacillary leprosy nor did itreduce serum ferritin levels.

Reduced glutathione accelerates the oxidative damage produced by sodium n-propylthiosulfate, one of the causative agents of onion-induced hemolytic anemia in dogs

The oxidative effects of sodium n-propylthiosulfate, one of the causative agents of onion-induced hemolytic anemia in dogs, were investigated in vitro using three types of canine erythrocytes, which are differentiated by the concentration of reduced glutathione and the composition of intracellular cations. After incubation with sodium n-propylthiosulfate, the methemoglobin concentration and Heinz body count in all three types of erythrocytes increased and a decrease in the erythrocyte reduced glutathione concentration was then observed. The erythrocytes containing high concentrations of potassium and reduced glutathione (approximately five times the normal values) were more susceptible to oxidative damage by sodium n-propylthiosulfate than were the normal canine erythrocytes. The susceptibility of the erythrocytes containing high potassium and normal reduced glutathione concentrations was intermediate between those of erythrocytes containing high concentrations of potassium and reduced glutathione and normal canine erythrocytes. In addition, the depletion of erythrocyte reduced glutathione by 1-chloro-2,4-dinitrobenzene resulted in a marked decrease in the oxidative injury induced by sodium n-propylthiosulfate in erythrocytes containing high concentrations of potassium and reduced glutathione. The generation of superoxide in erythrocytes containing high concentrations of potassium and reduced glutathione was 4.1 times higher than that in normal canine erythrocytes when the cells were incubated with sodium n-propylthiosulfate. These observations indicate that erythrocyte reduced glutathione, which is known as an antioxidant, accelerates the oxidative damage produced by sodium n-propylthiosulfate.

Effects of dietary cysteine on blood sulfur amino acid, glutathione, and malondialdehyde concentrations in cats

Abstract Objective To determine effects of dietary cysteine on blood sulfur amino acids (SAA), reduced glutathione (GSH), oxidized glutathione (GSSG), and malondialdehyde (MDA) concentrations in cats. Animals 12 healthy adult cats. Procedure Cats were fed diets with a nominal (0.50 g/100 g dry matter [DM]), moderate (1.00 g/100 g DM), or high (1.50 g/100 g DM) cysteine content in a 3 × 3 Latin square design with blocks of 8 weeks’ duration. Venous blood samples were collected after each diet had been fed for 4 and 8 weeks, and a CBC and serum biochemical analyses were performed; poikilocyte, reticulocyte, and Heinz body counts were determined; and MDA, GSH, GSSG, and SAA concentrations were measured. Results Blood cysteine and MDA concentrations were not significantly affected by dietary cysteine content. Blood methionine, homocysteine, and GSSG concentrations were significantly increased when cats consumed the high cysteine content diet but not when they consumed the moderate cysteine content diet, compared with concentrations obtained when cats consumed the nominal cysteine content diet. Blood GSH concentrations were significantly increased when cats consumed the moderate or high cysteine content diet. Conclusions Increased dietary cysteine content promotes higher blood methionine, homocysteine, GSH, and GSSG concentrations in healthy cats. Clinical Relevance Supplemental dietary cysteine may be indicated to promote glutathione synthesis and ameliorate adverse effects of oxidative damage induced by disease or drugs. (Am J Vet Res 1999;60:328–333)

Assessing the Neurotoxic Potential of Methyl Ethyl Ketoxime in Rats

Assessing the Neurotoxic Potential of Methyl Ethyl Ketoxime in Rats. SCHULZE, G. E., AND DERELANKO, M. J. (1993). Fundam. Appl. Toxicol. 21, 476–485. The potential of methyl ethyl ketoxime (MEKO) to produce neurotoxicity following acute and subchronic exposure was studed in rats. A Functional Observational Battery, assessment of motor activity, and neuropathology evaluations were conducted in the context of acute and subchronic toxicity studies. Three independent studies are reported: a pilot time-effect study designed to determine the time course and time to peak effect following a single high dose of MEKO, a single-dose neurotoxicity study, and a subchronic (13-week) repeated-dose neurotoxicity study in rats. An acrylamide-positive control group was included in the acute and subchronic studies for comparison with MEKO. Following an acute oral exposure of MEKO at a dose level of 900 mg/kg, locomotor activity was decreased compared to control with maximum decreases occurring between 30 and 60 min following oral administration. In the acute study, transient treatment-related changes in ease of cage removal, ease of handling, and in posture and gait were observed 1 hr after dosing with 900 mg/kg MEKO, as were significant depressions in motor activity. Following a single 300 mg/kg dose, transient MEKO-related changes in gait and aerial righting reflex were noted 1 hr after dosing. All effects were reversible within 24 hr of dosing. The single 100 mg/kg dose of MEKO was without observable effects. No acrylamide-related behavioral effects were noted following a single 50 mg/kg dose. In the subchronic study, transient treatment-related changes in ease of cage removal, ease of handling, and in posture, gait, and aerial righting were observed at the 400 mg/kg/day dose level when assessments were conducted immediately after dose administration. No consistent behavioral effects were observed prior to daily dose administration even after 13 weeks of exposure, indicating a lack of cumulative behavioral effect. No consistent behavioral changes were noted at doses of 125 mg/kg/day and below. Significant dose-related decreases in red cell mass, and increases in methemoglobin levels, reticulocyte, leukocyte, Heinz body counts, and spleen weights occurred at subchronic MEKO doses of 40 mg/kg/day and higher. No MEKO-related neuropathological changes occurred. Animals receiving acrylamide at 20 mg/kg/day showed expected behavioral and neuropathological changes consistent with peripheral neuropathy. In conclusion, high doses of MEKO can produce transient and reversible changes in neurobehavioral function consistent with central nervous system (CNS) depression. No evidence of cumulative neurotoxicity was detected. The hematopoietic system was effected at doses which did not produce detectable changes in CNS function.

Assessing the Neurotoxic Potential of Methyl Ethyl Ketoxime in Rats

Assessing the Neurotoxic Potential of Methyl Ethyl Ketoxime in Rats. Schulze, G. E., and Derelanko, M. J. (1993). Fundam. Appl. Toxicol. 21, 476-485.The potential of methyl ethyl ketoxime (MEKO) to produce neurotoxicity following acute and subchronic exposure was studied in rats. A Functional Observational Battery, assessment of motor activity, and neuropathology evaluations were conducted in the context of the acute and subchronic toxicity studies. Three independent studies are reported: a pilot time-effect study designed to determine the time course and time to peak effect following a single high dose of MEKO, a single-dose neurotoxicity study, and a subchronic (13-week) repeated-dose neurotoxicity study in rats. An acrylamide-positive control group was included in the acute and subchronic studies for comparison with MEKO. Following an acute oral exposure of MEKO at a dose level of 900 mg/kg, locomotor activity was decreased compared to control with maximum decreases occurring between 30 and 60 min following oral administration. In the acute study, transient treatment-related changes in ease of cage removal, ease of handling, and in posture and gait were observed 1 hr after dosing with 900 mg/kg MEKO, as were significant depressions in motor activity. Following a single 300 mg/kg dose, transient MEKO-related changes in gait and aerial righting reflex were noted 1 hr after dosing. All effects were reversible within 24 hr of dosing. The single 100 mg/kg dose of MEKO was without observable effects. No acrylamide-related behavioral effects were noted following a single 50 mg/kg dose. In the subchronic study, transient treatment-related changes in ease of cage removal, ease of handling, and in posture, gait, and aerial righting were observed at the 400 mg/kg/day dose level when assessments were conducted immediately after dose administration. No consistent behavioral effects were observed prior to daily dose administration even after 13 weeks of exposure, indicating a lack of cumulative behavioral effect. No consistent behavioral changes were noted at doses of 125 mg/kg/day and below. Significant dose-related decreases in red cell mass, and increases in methemoglobin levels, reticulocyte, leukocyte, Heinz body counts, and spleen weights occurred at subchronic MEKO doses of 40 mg/kg/day and higher. No MEKO-related neuropathological changes occurred. Animals receiving acrylamide at 20 mg/kg/day showed expected behavioral and neuropathological changes consistent with peripheral neuropathy. In conclusion, high doses of MEKO can produce transient and reversible changes in neurobehavioral function consistent with central nervous system (CNS) depression. No evidence of cumulative neurotoxicity was detected. The hematopoietic system was effected at doses which did not produce detectable changes in CNS function.

Effects of oral administration of brassica secondary metabolites, allyl cyanide, allyl isothiocyanate and dimethyl disulphide, on the voluntary food intake and metabolism of sheep

Glucosinolates, such as sinigrin, and S-methyl cysteine sulphoxide (SMCO), which are found in forage brassica species have been implicated in the low intakes observed among lambs consuming such diets. To test both the individual and interactive effects of these compounds in sheep, all combinations of the sinigrin breakdown products, allyl cyanide (ACN) and allyl isothiocyanate (AITC; 10 mmol/d), and the SMCO metabolite dimethyl disulphide (DMDS; 25 mmol/d) were orally administered twice daily for 5 weeks to forty sheep offered dried grass pellets ad lib. As well as measuring voluntary food intake (VFI), a number of haematological and clinical function tests were conducted to assess the physiological effects of the compounds. VFI was significantly depressed by both ACN and AITC but not by DMDS. DMDS significantly ameliorated the effects of ACN on VFI (P < 0.001). Concentrations of reduced glutathione in the blood were depressed by ACN and AITC and elevated by DMDS but no significant interactions were evident. Elevated plasma gamma-glutamyl transpeptidase (EC 2.3.2.1) activity on ACN and AITC treatments indicated possible liver damage. DMDS elicited a rise in Heinz bodies to 11% by week 2 but this was not reflected in packed cell volume and blood haemoglobin levels which were unaffected by treatment. The increased Heinz body count caused by DMDS was not further influenced by ACN or AITC. In conclusion, the depressive effects of sinigrin breakdown products on VFI were not compounded by the additional presence of DMDS which, on the contrary, lessened the depression of VFI caused by ACN.

Elevated Glutathione Accelerates Oxidative Damage to Erythrocytes Produced by Aromatic Disulfide

It has been shown that certain dogs have erythrocytes characterized by an inherited high concentration of reduced glutathione (GSH), five to seven times the normal level (high-GSH RBCs). We examined whether increased GSH in dog erythrocytes leads to increased protection against oxidative damage induced by acetylphenylhydrazine (APH) and/or 4-aminophenyl disulfide (4-AD). When erythrocytes were incubated with 30 mmol/L APH, the Heinz body count was appreciably higher in normal RBCs than in high-GSH RBCs, while there was no difference in the increase of the methemoglobin (metHb) concentration in both RBCs. In contrast, both the Heinz body count and metHb production were much higher in high-GSH RBCs than in normal RBCs when erythrocytes were incubated with 4-AD. Furthermore, the generation of the superoxide in erythrocytes treated with 4-AD, which was measured by spin trapping combined with electron spin resonance (ESR), was obviously higher in high-GSH RBCs than in normal RBCs. These results clearly indicate that erythrocyte GSH is an important defense against oxidative damage induced by certain compounds such as APH, but that, in contrast, elevated GSH appears to accelerate oxidative damage to erythrocytes produced by aromatic disulfides, such as 4-AD, which generated a superoxide in erythrocytes via its redox reaction with GSH.

Elevated glutathione accelerates oxidative damage to erythrocytes produced by aromatic disulfide

It has been shown that certain dogs have erythrocytes characterized by an inherited high concentration of reduced glutathione (GSH), five to seven times the normal level (high-GSH RBCs). We examined whether increased GSH in dog erythrocytes leads to increased protection against oxidative damage induced by acetylphenylhydrazine (APH) and/or 4- aminophenyl disulfide (4-AD). When erythrocytes were incubated with 30 mmol/L APH, the Heinz body count was appreciably higher in normal RBCs than in high-GSH RBCs, while there was no difference in the increase of the methemoglobin (metHb) concentration in both RBCs. In contrast, both the Heinz body count and metHb production were much higher in high-GSH RBCs than in normal RBCs when erythrocytes were incubated with 4-AD. Furthermore, the generation of the superoxide in erythrocytes treated with 4-AD, which was measured by spin trapping combined with electron spin resonance (ESR), was obviously higher in high-GSH RBCs than in normal RBCs. These results clearly indicate that erythrocyte GSH is an important defense against oxidative damage induced by certain compounds such as APH, but that, in contrast, elevated GSH appears to accelerate oxidative damage to erythrocytes produced by aromatic disulfides, such as 4-AD, which generated a superoxide in erythrocytes via its redox reaction with GSH.

Heinz body anaemia in lambs with deficiencies of copper or selenium.

1. The progression of Heinz body anaemia was studied in groups of lambs of low- and high-copper status, produced through breeding or Cu supplementation, when they were transferred from improved pasture to rape (Brassica napus L.) in autumn. Some lambs had previously received selenium by injection. The Cu and Se supplements markedly increased superoxide dismutase (EC 1.15.1.1; SOD) and glutathione peroxidase (EC 1.11.1.9; GSHPx) activities respectively in the erythrocytes, and both supplements had elicited growth responses at pasture. 2. At the time of transfer to rape, lambs not treated with Cu had lower whole-blood haemoglobin (Hb) concentrations and a higher percentage of erythrocytes containing Heinz bodies (6.6 v. 3.7%, P less than 0.01) than Cu-treated lambs: the corresponding effects of Se treatment were similar in direction but lower in magnitude (P less than 0.05). 3. After grazing rape for 2 weeks the mean Hb concentration had fallen by 30 g/l while Heinz body count had increased from 5 to 25%. However, counts were negatively correlated with the initial values and were unaffected by the Cu and Se treatments which maintained high plasma Cu concentrations and SOD and GSHPx activities. 4. The results provide the first evidence that Cu deficiency can induce Heinz body formation and the anaemia in grazing Cu-deficient lambs may be partly haemolytic in origin. The concomitant Se deficiency added marginally to the problem but neither the separate nor combined deficiencies increased the susceptibility of lambs to brassica anaemia.

Subchronic oral toxicology of 4-chloro-3-nitroaniline in the rat

4-Chloro-3-nitroaniline was given to groups of 20 male and 20 female rats by gavage at doses of 3.6, 18, or 90 mg/kg in a 10% corn oil suspension. Doses were administered 5 days per week for 90 days. The high dose resulted in reduced body weight gain in males, reduced feed consumption and fluctuating feed consumption in both sexes, slight hemolytic anemia with Heinz bodies in both sexes, enlargement of the spleen due to congestion, hemosiderosis, and increased extramedullary hematopoiesis in both sexes, inflammatory changes in the splenic capsules of two females, hemosiderin pigmentation of the liver in both sexes, bone marrow hyperplasia in both sexes, increased liver weight in females, and testicular atrophy. The middle dose produced a fluctuating feed consumption pattern in males, Heinz bodies in both sexes, very slight anemia in females, splenic hemosiderosis in males, slightly increased splenic extramedullary hematopoiesis in females, hemosiderin pigmentation of the liver in males, and possibly increased liver weight in females, and inflammatory changes in the splenic capsule of one male. The low dose produced Heinz bodies in males and possibly females although the Heinz body counts of females were not statistically significant. The primary toxicologic damage produced by 4-chloro-3-nitroaniline is hemolytic or Heinz body anemia and testicular atrophy. Thus the toxicity of 4-chloro-3-nitroniline is similar to that of other aromatic nitro and amino chemicals.

Effect of onion ingestion on anti-oxidizing agents in dog erythrocytes.

The effect of onion ingestion on red cell anti-oxidizing agents such as superoxide dismutase (SOD), catalase, glucose-6-phosphate dehydrogenase (G-6-PD), glutathione peroxidase (GPx), NADH-methemoglobin reductase and reduced glutathione (GSH) was studied in relation to Heinz body hemolytic anemia in dogs. To dogs was given the onion soup equivalent to 30g of raw onion/kg body weight once a day for three days. A few days after the initiation of feeding, there was a transient decrease of SOD, catalase and G-6-PD activities to 60%, and GSH value to 70% of their predosing values with a sudden and marked Heinz body formation. These values then began to increase when Heinz bodies were observed in almost every red cells. On day 10, when red cells carrying Heinz bodies were almost cleared from blood circulation, catalase and G-6-PD reached peak values of nearly 250% of their predosing levels. Changes of GPx and NADH-methemoglobin reductase were not correlated with Heinz body counts. Methemoglobin concentration was significantly elevated and maintained in the level of 2-3% during the presence of Heinz bodies in the red cells.

Subchronic Oral Toxicology of 4-Chloro-3-nitroaniline in the Rat

4-Chloro-3-nitroaniline was given to groups of 20 male and 20 female rats by gavage at doses of 3.6, 18, or 90 mg/kg in a 10% corn oil suspension. Doses were administered 5 days per week for 90 days. The high dose resulted in reduced body weight gain in males, reduced feed consumption and fluctuating feed consumption in both sexes, slight hemolytic anemia with Heinz bodies in both sexes, enlargement of the spleen due to congestion, hemosiderosis, and increased extramedullary hematopoiesis in both sexes, inflammatory changes in the splenic capsules of two females, hemosiderin pigmentation of the liver in both sexes, bone marrow hyperplasia in both sexes, increased liver weight in females, and testicular atrophy. The middle dose produced a fluctuating feed consumption pattern in males, Heinz bodies in both sexes, very slight anemia in females, splenic hemosiderosis in males, slightly increased splenic extramedullary hematopoiesis in females, hemosiderin pigmentation of the liver in males, and possibly increased liver weight in females, and inflammatory changes in the splenic capsule of one male. The low dose produced Heinz bodies in males and possibly females although the Heinz body counts of females were not statistically significant. The primary toxicologic damage produced by 4-chloro-3-nitroaniline is hemolytic or Heinz body anemia and testicular atrophy. Thus the toxicity of 4-chloro-3-nitroaniline is similar to that of other aromatic nitro and amino chemicals.

Vitamin E and Dapsone-Induced Hemolysis

Sixteen patients, each receiving 100 mg of dapsone per day, were studied for evidence of hemolysis. Vitamin E (dl-alpha tocopherol acetate), 800 mg/day, was then administered for up to three months, and dapsone therapy was continued at the same dose. Hemolysis factors were reexamined immediately prior to cessation of vitamin E therapy. No substantial change was demonstrable for levels of hemoglobin, reticulocyte count, and haptoglobin at the end of vitamin E therapy, despite a significant rise in serum vitamin E levels. Erythrocyte survival measured in four patients before and at the end of vitamin E therapy also showed no substantial change. Erythrocyte Heinz body count, however, fell in nine of 15 patients studied, and none showed an increase in this measurement while receiving vitamin E. We conclude that in patients receiving dapsone at 100 mg/day, vitamin E therapy at 800 mg/day does not substantially ameliorate the hemolytic effect of this drug.

Abnormal erythrocyte metabolism in hepatic disease

Erythrocyte (RBC) metabolic studies were done on 114 patients with severe hepatic disease. Heinz body formation after incubation of RBCs with acetyl phenylhydrazine was found to be significantly higher in patients than in controls. RBC-reduced glutathione levels were lower than those of controls both before and after incubation with acetyl phenylhydrazine, and patients with the highest Heinz body counts had the lowest reduced glutathione levels. RBC methylene blue-stimulated hexose monophosphate (HMP) shunt metabolism and glucose recycling through the shunt were significantly lower in patients with active hepatic disease than in controls. There was no difference in resting HMP shunt activity or in resting recycling of glucose. Despite impairment of shunt metabolism, total glucose consumption was greater in patients than in controls. The patients with the lowest stimulated HMP shunt metabolism and glucose recycling had the highest Heinz body counts, lowest reduced glutathione, and highest total glucose consumption. A continuum of abnormal shunt metabolism was seen, from a mild reduction of stimulated HMP shunt activity to a severe combined decrease in both the HMP shunt and glucose recycling. When measured, glutathione reductase, glutathione peroxidase, glucose-6-phosphate dehydrogenase, and transketolase were normal or increased. Sequential studies were done on 11 patients who had abnormal metabolic studies. Coincident with improvement of HMP shunt metabolism, the Heinz body counts became lower, reduced glutathione higher, hematocrit higher, and liver function improved. Impaired HMP shunt metabolism appears to be a common, acquired RBC abnormality in patients with severe, active liver disease.

Abnormal Erythrocyte Metabolism in Hepatic Disease

Erythrocyte (RBC) metabolic studies were done on 114 patients with severe hepatic disease. Heinz body formation after incubation of RBCs with acetyl phenylhydrazine was found to be significantly higher in patients than in controls. RBC-reduced glutathione levels were lower than those of controls both before and after incubation with acetyl phenylhydrazine, and patients with the highest Heinz body counts had the lowest reduced glutathione levels. RBC methylene blue-stimulated hexose monophosphate (HMP) shunt metabolism and glucose recycling through the shunt were significantly lower in patients with active hepatic disease than in controls. There was no difference in resting HMP shunt activity or in resting recycling of glucose. Despite impairment of shunt metabolism, total glucose consumption was greater in patients than in controls. The patients with the lowest stimulated HMP shunt metabolism and glucose recycling had the highest Heinz body counts, lowest reduced glutathione, and highest total glucose consumption. A continuum of abnormal shunt metabolism was seen, from a mild reduction of stimulated HMP shunt activity to a severe combined decrease in both the HMP shunt and glucose recycling. When measured, glutathione reductase, glutathione peroxidase, glucose-6-phosphate dehydrogenase, and transketolase were normal or increased. Sequential studies were done on 11 patients who had abnormal metabolic studies. Coincident with improvement of HMP shunt metabolism, the Heinz body counts became lower, reduced glutathione higher, hematocrit higher, and liver function improved. Impaired HMP shunt metabolism appears to be a common, acquired RBC abnormality in patients with severe, active liver disease.