Effect Of Isonicotinic Acid Hydrazide Research Articles

Low Cytosolic Non-Heme Iron Levels in Erythroid Cells Prevent IRP2-Mediated Ferritin Upregulation during Differentiation.

Erythroid cells are the major consumers of iron in the human body. Differentiating erythroid cells shuttle the metal with very high efficiency towards mitochondria for the formation of heme. To satisfy their high iron needs, developing red blood cells (RBC) have to sustain high expression of transferrin receptor 1 (TfR) despite increasing cellular iron concentration. Moreover, synthesis of ferritin must not be activated by incoming iron, since this would represent a counterproductive storage during the phase of high iron demand. Recently we have demonstrated that during terminal differentiation primary erythroid cells satisfy their exceptionally high requirements for iron by switching to a mode where the post-transcriptional, iron-dependent regulatory system, formed by iron responsive proteins (IRP1 and IRP2) and iron responsive elements (IREs), seems to sense a low-iron state. This occurs despite a massive net increase of iron import into the cell (Schranzhofer et al., Blood 107:4159, 2006). To examine the hypothesis that erythroid cells have low non-heme iron levels in their cytosol, we experimentally increased the cytosolic iron pool by either inhibiting heme biosynthesis or overloading cells with iron. Both block of heme synthesis by either succinylacetone or isonicotinic acid hydrazide (INH) or administration of ferric ammonium citrate, resulted in a clear increase in ferritin levels. This increase was directly proportional to the increase in the cellular concentration of non-heme-iron. Moreover, the effect of INH, the inhibitor of 5-aminolevulinic acid (ALA) synthase, could be reversed by the addition of ALA. Strikingly, increases in ferritin expression upon perturbation of cellular iron homeostasis strongly correlated with the loss of IRE-binding activity of IRP2 but not IRP1, as determined by mobility shift assays. This suggests that IRP2 is the major regulator of ferritin expression in erythroid cells. To further elaborate on this observation, we cultured primary erythroblasts derived from IRP1−/− and IRP2−/− mice (kindly provided by Drs. M. Hentze and B. Galy). In agreement with the published phenotype of microcytic hypochromic anemia, only erythroblasts lacking IRP2 exhibited a reduction in hemoglobinization. Moreover, only IRP2−/− cells showed a significant increase in ferritin expression, whereas developing RBC lacking IRP1 had levels of ferritin protein equal to wild type cells. We conclude that in erythroid cells efficient shuttling of incoming iron towards mitochondria and its prompt use for heme formation is important to keep the cytosol in an iron-deprived state and consequently ferritin protein levels low. This translational repression seems to be mainly achieved by IRP2. Together with the observation that surface expression of TfR was reduced in IRP2−/− erythroblasts during self renewal but not during terminal differentiation, our results suggest that not only down-regulation of TfR, but also up-regulation of ferritin may be a major factor for the anemic phenotype observed in IRP2−/− mice.

Isonicotinic acid hydrazide: an anti-tuberculosis drug inhibits malarial transmission in the mosquito gut

We studied the transmission-blocking effect of isonicotinic acid hydrazide (INH), a widely used anti-tuberculosis drug, against Plasmodium gallinaceum and Plasmodium berghei. INH-treatment of infected animals did not inhibit parasite development in the blood of the vertebrate host, but did inhibit exflagellation, ookinete formation, and oocyst development in the mosquito. Oocyst development was inhibited in a dose-dependent manner. The ED 50 in the P. gallinaceum/chicken/ Aedes aegypti model and P. berghei/mouse/ Anopheles stephensi model was 72 and 109 mg/kg, respectively. In marked contrast, in vitro exflagellation and ookinete development were not directly affected by physiological concentrations of INH. We suggest that INH exerts its inhibitory effects on the mosquito stages of the malaria parasite by an indirect, and at present undefined mechanism. Further elucidation of the mechanism how INH inhibits parasite development specifically on mosquito stages may allow us to identify new targets for malaria control strategy. Index Descriptors and Abbreviations: Plasmodium berghei; Plasmodium gallinaceum; Anopheles stephensi; Aedes aegypti; Transmission-blocking anti-malarial; INH, isonicotinic acid hydrazide; XA, xanthurenic acid; GAF, gametocyte activating factor; HBSA, Hepes-buffered suspended animation medium; EI, exflagellation index; ED 50, half-maximal effective dose

In vivo inhibition of kynurenine aminotransferase activity by isonicotinic acid hydrazide in rats.

It is known that the anti-tuberculosis drug, isonicotinic acid hydrazide (INH), causes pellagra, a niacin deficiency syndrome, and peripheral neuritis in humans. We investigated the effects of INH on the metabolism of tryptophan to niacin in rats fed on a niacin-free diet. The activity of kynurenine aminotransferase was significantly inhibited by feeding a diet containing INH and by an injection of INH, and the urinary excretion of xanthurenic acid, the side-reaction product of the conversion pathway of tryptophan to niacin, was below the limit of detection. The inhibition of kynurenine aminotransferase and the resulting decreased formation of xanthurenic acid generally mean a higher conversion ratio of tryptophan to niacin. However, the conversion ratio was no different between the control and INH groups.

Effect of Isonicotinic Acid Hydrazide on Extracellular Amino Acids and Convulsions in the Rat: Reversal of Neurochemical and Behavioural Deficits by Sodium Valproate

We have studied the effect of isonicotinic acid hydrazide (INH), a convulsant agent, on the extracellular levels of amino acids in the hippocampus, and the effect of sodium valproate (VPA) administration in INH-treated rats. INH (250 mg/kg) caused a rapid and sustained decrease in basal levels of GABA, and during this period convulsions of increasing severity were observed. Basal levels of glutamine, taurine, aspartate, and glutamate were unchanged by INH. When VPA was coadministered with INH, basal GABA levels were increased and no convulsions were observed. When transmitter release was evoked using 100 mM K+, the increase in dialysate GABA observed in INH-treated animals was less than that seen in controls and convulsions increased in frequency. K(+)-evoked release of glutamate and aspartate tended to be higher following INH treatment, and in the case of aspartate, this increase was significant. VPA reversed the changes in evoked release of glutamate and aspartate, and release of GABA was considerably greater than that seen in control or INH-treated rats. No drug effect on evoked changes in taurine or glutamine level was seen. These are the first data to show decreased extracellular GABA in conjunction with convulsions in freely moving animals in vivo.

The Synergistic Effect of Isonicotinic Acid Hydrazide (INH) and Griseofulvin (GF) on Porphyrin Metabolism

Abnormal porphyrin metabolism can be induced by several chemicals. To investigate the synergistic effect on porphyrinopathy of isonicotinic acid hydrazide (INH) with a low concentration of griseofulvin (GF), the two chemicals were given to mice simultaneously. INH was added to drinking water at a concentration of 0.05%. GF was mixed with feed at a concentration of 0.1%. The mice (dd-y strain) were divided into four groups. Those in group A were fed normally. Group B received only 0.1% GF, group C received only 0.05% INH, and group D received both 0.1% GF and 0.05% INH. The treatment was continued for 13 to 30 weeks. After treatment, the mice were anesthetized and sacrificed. The liver and whole blood were taken for analysis of porphyrins. The results revealed a slight elevation of erythrocytic porphyrins in the groups treated with 0.1% GF or 0.05% INH alone and remarkable abnormalities in the hepatic and erythrocytic porphyrin levels of the group simultaneously treated with both chemicals. These results show that INH itself may have a small potential for the induction of porphyric abnormalities, and that the administration of INH with a low concentration of GF induces marked porphyrinopathy in dd-y strain mice.

Effect of isoniazid, a haem inhibitor, on globin chain synthesis in reticulocytes from non-thalassaemic and beta thalassaemic subjects.

The effect of isonicotinic acid hydrazide (INH), a potent haem inhibitor, on globin chain synthesis was studied in reticulocytes from the following groups of patients: four non-thalassaemic patients (group i); five beta thalassaemia heterozygotes (group ii); three Hb S/beta thalassaemia heterozygotes (group iii); and two additional patients--one with homozygous beta thalassaemia and the other with thalassaemia intermedia (group iv). This was done to determine whether haem inhibitors depress alpha globin chain synthesis. The progressive increase of INH concentration (10-40 mmol l-1) in reticulocytes from a beta thalassaemia heterozygote resulted in a remarkable decrease of the alpha and beta chain synthesis, ranging from 80% to 97% and from 74% to 96% of control values, respectively, and in a gradual drop of alpha:beta ratio from 1.87 to 1.38. Furthermore, in the samples incubated with 40 mmol l-1 INH, a pronounced inhibition of globin chain synthesis 77 (19%) for alpha chain and 67 (27%) for beta or beta S chain) and a substantial drop of the alpha:beta or beta S ratio in samples with INH (median 1.16) compared with that in samples without INH (median 1.70) were observed. The inhibitory effect of INH was significantly or completely corrected by adding exogenous haem. It is suggested that haem inhibition and the resulting preferential diminution of alpha chain synthesis could provide a new approach to the treatment of homozygous beta thalassaemia with an excess of detrimental free alpha chain in erythroid cells.

Porphyrinostatic effect of isonicotinic acid hydrazide on porphyric chick embryo and yeast.

1. The common feature of "inducible" forms of hepatic porphyria is an increased activity of delta-amino-levulinate/ALA/synthase. 2. As a potential therapeutic procedure, isonicotinic acid hydrazide/INH/as inhibitor of this enzyme, was applied and found to show a porphyrinostatic effect, depending on its concentration, both in yeast and porphyric chick embryo models. 3. Moreover, pyridoxalphosphate/PDX/eliminated the porphyrinostatic effect of INH, depending again on its concentration. 4. Histochemical tests did not demonstrate any significant functional or morphological changes in the chick embryo hepatocytes following INH administration.

Drug metabolism in experimental tuberculosis: II. Modification of monooxygenase activities due to infection by the administration of isonicotinic acid hydrazide.

Isonicotinic acid hydrazide (INH) upon administration to tuberculous guinea pigs exhibited curative effect by bringing back the tissue weights to normal. The reduced hepatic content of cytochrome P450 and NADPH-cytochrome C reductase activity of tuberculous guinea-pigs was restored to normal by INH treatment. Also a similar effect of INH was observed in the case of aminopyrine-N-demethylase and benzphetamine-N-demethylase in the liver and lung of tuberculous guinea-pigs. But INH had little effect on the content of cytochrome b5 and NADH-cytochrome b5 reductase activity. The hepatic and pulmonary activities of glutathione-S-transferase exhibited a tendency to come back to normal while UDP-glucuronyl transferase was unaffected by INH administration to the infected animals. The results suggest that the treatment of tuberculous guinea-pigs with INH cause the normalisation of disturbed hepatic and pulmonary metabolism.

The effect of substrate on inhibition of Corynebacterium lepus by isonicotinic acid hydrazide (Isoniazid)

Isonicotinic acid hydrazide (INH) inhibits the growth of Corynebacterium lepus on hexadecane but has no effect on its growth on fructose. INH also inhibits the production of the mycolic acid containing lipopeptide bioemulsifier normally produced by C. lepus in response to an insoluble substate. The primary effect of INH appears to be inhibition of mycolic acid synthesis, which limits the growth of C. lepus on hexadecane by reducing the concentration of bioemulsifier.

Effect of isonicotinic acid hydrazide on the intratumor injection of BCG

Effect of isonicotinic acid hydrazide on the intratumor injection of BCG

Effect of Isonicotinic Acid Hydrazide on the Intratumor Injection of BCG2

Rats with established subcutaneous tumors were treated by repeated intratumor injections of BCG over a 5-week period. Isonicotinic acid hydrazide (INH) was given to some rats to determine if this drug decreased the effect of BCG on local tumor growth. INH alone had no effect on either survival or tumor volume. Rats treated with either BCG or BCG and INH had prolonged survivals and smaller tumor volumes than did controls rats. In these experiments, INH did not decrease the effect of BCG on local tumor growth. However, the intratumor injection of BCG in rats receiving INH systematically was associated with better survival and smaller tumor volumes compared to the intratumor injection of BCG alone.

N-bromosuccinimide protects chick embryos against the lathyrogenic effect of isonicotinic acid hydrazide.

SummaryIsonicotinic acid hydrazide when applied to the chorio‐allantoic membrane of 14 days old chick embryos produced after 3 days of further incubation a marked increase in the fragility of the embryo tissues. It also increased the amount of neutral salt‐soluble collagen. The administration of N‐Bromo‐succinimide reversed this effect and at the same time increased the elasticity of the embryo above the normal control. It is suggested that NBS acts by virtue of its aldehyde group.ZusammenfassungN‐Bromosuccinimid schützt Hühnerembryonen gegen den lathyrogenen Effekt von Isonikotinsäure‐HydrazidDie Applikation von Isonikotinsäure‐Hydrazid in die Membran des Allantochorions von 14 Tage alten Hühnerembryonen erhöht innerhalb 3 Tagen die Fragilität der embryonalen Gewebe. Die Verabreichung von N‐Bromosuccinimid (NBS) macht dem Effekt räckgängig, wobei die Gewebselastizität sogar über den von normalen Kontrolltieren ansteigt. NBS wirkt wahrscheinlich dank seiner Aldehydgruppe.RésuméAction protectrice de la N‐bromosuccinimide contre l'effet lathyrogène de l'hydrazide de l'acide isotonique sur les embryons de pouletsSous l'action de l'hydrazide de l'acide isonicotinique sur la membrane allanto‐choriale des embryons de poulets âgés de 14 jours, il se produit, après 3 jours d'incubation, une nette augmentation de la fragilité des tissus embryonnaires. II se produit en outre une augmentation de la quantité des collagènes neutres solubles dans les sels. L'administration de la N‐bromosuccinimide produit un phénomène inverse et augmente en même temps l'élasticité de l'embryon au‐delà de celle du témoin normal. On suggère que la N‐bromosuccinimide pourrait agir par son groupement aldéhydique.ResumenLa protección de embriones de pollo con N‐bromosuccinimida frente al efecto latirógeno de la hidrazida del ácido isonicotínicoLa aplicación de hidrazida del ácido isonicotínico en la membrana corioalantoidea de embriones de pollo de 14 días de edad aumenta en un plazo de 3 días la fragilidad de los tejidos embrionarios. También aumenta la cantidad de colágeno neutro soluble en sal. La administración de N‐bromosuccinimida (NBS) hace reversible este efecto, aumentando la elasticidad hística incluso por encima de la de los animales testigos normales. NBS actúa probablemente gracias a su grupo aldehido.

Sustained drug-induced elevation of brain GABA in the rat.

Abstract— The contents of GABA, homocarnosine, and β‐alanine can be raised in rat brain for long periods of time by the continued administration of phenelzine, aminooxyacetic acid (AOAA), or isonicotinic acid hydrazide (INH). These 3 compounds apparently act by preferential inhibition of the enzyme GABA aminotransferase (GABA‐T). Oral administration of phenelzine (20 mg/kg per day) caused a 25–50 per cent increase in GABA levels in rat brain, but produced appreciable toxic side effects. A similar increase in GABA levels in brain resulted from oral administration to rats of INH in a dosage of 60 mg/kg per day, without production of any obvious toxic effects. Simultaneous administration of large doses of pyridoxine did not abolish the GABA‐elevating effect of INH. Brain GABA levels in the rat were increased by approx. 50 per cent by daily injections of AOAA (2.5 mg/kg per day). At this low dosage, AOAA injections in rats could be continued for at least 6 weeks without producing evident toxic effects. Oral administration of large amounts of GABA, on the other hand, failed to increase the content of GABA in the brains of rats not treated with GABA‐T inhibitors, and failed to produce any further increase of brain GABA levels in rats treated with AOAA.

Tumor regression mediated by Mycobacterium bovis (strain BCG). Effects of isonicotinic acid hydrazide, cortisone acetate, and antithymocyte serum.

Tumor regression mediated by Mycobacterium bovis (strain BCG). Effects of isonicotinic acid hydrazide, cortisone acetate, and antithymocyte serum.

The effects of isonicotinic acid hydrazide on the early chick embryo

ABSTRACT Unincubated White Leghorn fertile eggs were injected with isonicotinic acid hydrazide (INH) through a hole in the shell. A control group was injected with normal saline, a second group with INH plus vitamin B6, and a third group was left unopened. INH, in the doses we used, proved to be lethal for more than 50% of the early chick embryos, and also produced important developmental alterations. Alterations produced by INH were primarily observed at the level of the neural epithe-lium, particularly at its cephalic portion. The most important ones were a degenerative and necrotic process of the neural epithelium, and a distortion of the normal anatomical relations of the cephalic structures. The embryonic mortality and developmental alterations induced by INH were prevented to a considerable extent by the concurrent injection of pyridoxine hydrochloride and INH. The preventive action of vitamin B6 suggests that toxicity of INH in the chicken embryos is due to the antivitamin-B6 action of INH. Through this mechanism INH would block, specially, the amino acid metabolism. The developing nervous system was the embryonic area most sensitive to such metabolic alterations.

Lack of carcinogenic effects of isonicotinic acid hydrazide in the Syrian golden hamsters.

Isonicotinic acid hydrazide (INH) was administered as a 0.3 % solution for 42 weeks intermittently and as a 0.2 and 0.1 % solutions continuously in the drinking water for the lifespan of adult Syrian golden hamsters. 0.3 % INH solution was found to be too toxic, therefore it had to be given periodically and finally the treatment had to be stopped, while 0.2 and 0.1 % INH solutions were relatively well tolerated during the entire life of the animals. The consumption of INH resulted in no detectable tumorigenic effect in the treated hamsters. It is concluded that the drug is apparently no carcinogenic in this species. In view of the fact that the tumor producing capability of INH was shown by numerous studies in mice, the matter of selective neoplastic response in the various species is discussed and emphasized.

Nuclear and microsomal nicotinamide-adenine dinucleotidases of rat liver

1. 1. Nicotinamide-adenine dinucleotidase activity is present in both the nuclear and microsomal fractions of rat liver homogenates. 2. 2. Comparison of the nicotinamide-adenine dinucleotidase activity of a partially purified nuclear fraction with that of the microsomal fraction revealed that the two differed in apparent K m values, ratios of maximum velocities with nicotinamide-adenine dinucleotide and nicotinamide-adenine dinucleotide phosphate, pH optima, solubility and effects of isonicotinic acid hydrazide and pyridine. 3. 3. It is concluded that the nicotinamide-adenine dinucleotidases associated with the nuclear and microsomal fraction of rat liver are different enzymes.

The effect of isonicotinic acid hydrazide on the development of tumors.

AbstractIsonicotinic acid hydrazide (INH) was administered as a 0.1% solution in the drinking water for the life span of adult AKR inbred mice. The treatment resulted in no significant inhibitory effect on the development of malignant lymphomas which are characteristic of these animals, nor did it induce other tumors.Similar administration of INH as 0.05 and 0.15% solutions to adult randomly bred Swiss mice enhanced the formation of pulmonary adenomas, while it had no apparent inhibitory influence on the progression of other types of tumors.Single doses of 500, 400, 300, 250, 200 and 100 μg of INH were injected subcutane‐ously on the day of birth to Swiss mice, without significant tumorigenic effect.The problem of tumor induction versus inhibition is discussed.

Effect of isoniazid on biosynthesis in Mycobacterium tuberculosis var. bovis BCG.

Surmmary Radioactive incorporation techniques have been employed to investigate the effect of isonicotinic acid hydrazide (INH) on biosynthetic reactions in Mycobacterium tuberculosis var. bovis BCG. INH has no measurable effect on incorporation of 32P phosphate, 35S sulphate or 14C glycine into whole cells, although it inhibits by 50% the incorporation of 14C acetate. Incorporation of 32P into trichloracetic acid (TCA) insoluble components of supernatant solutions from centrifuged disrupted bacteria appears to be inhibited soon after the cells are exposed to INH. Similar experiments with 35S show a lag of 30–40 hr before incorporation is influenced by the drug. Chemical fractionation of organisms grown in the presence of 32P show that the major effect of INH is exerted on incorporation into the ribonucleic acid fraction.

Tumor-inducing effect of hydrazine in mice.

AFTER the discovery of the experimental tumour-inducing effect of isonicotinic acid hydrazide1 several later investigations verified that this compound showed carcinogenic effects in white mice of different inbred strains when administered intraperitoneally or orally over a long period2–8. According to Mori et al.9, the carbamyl group is responsible for the cancerogenic effect of isonicotinic acid hydrazide. On the other hand, Biancifiori et al.3 are of the opinion that the hydrazine radical10 detached from the compound during the metabolic process plays a part in producing tumours. They found in their experiments that the frequency of tumours in mice of BALB/c strain caused by isonicotinic acid hydrazide or hydrazine sulphate was very high, while isonicotinic acid alone showed no carcinogenic effect.