ACTA MEDICA (HRADEC KRÁLOVÉ)
1997, Vol. 40, No. 1 CONTENTS ORIGINAL ARTICLES
Vladimír Geršl, Radomír Hrdina, Jaroslava Vávrová, Vladimír Palička, Jaroslava Voglová, Yvona Mazurová, Jiří Bajgar Effects of repeated administration of dithiol chelating agent - sodium 2,3-dimercapto-1-propanesulphonate (DMPS)
- on biochemical and haematological parameters in rabbits ... 3
Josef Herink
Effect of alprazolam and ketamine on seizures induced by two different convulsants ... 9
Dušan Šimkovič, Milan Široký, Karel Šmejkal
The chronic anal fissure treatment by internal partial lateral sphincterotomy and following anal manometry ... 13
BRIEF COMMUNICATION
Eliška Marklová, Miluše Brátová, Viktor Voříšek
Screening for organic acid disorders ... 17
CASE REPORT
Jan Harrer, Miroslav Brtko, Pavel Žáček, Ján Knap
Hemothorax - a complication of subclavia vein connulation ... 21
HISTORICAL ARTICLE
Ivan Hybášek
The history of otorhinolaryngology at the Charles University, faculty of medicine in Hradec Králové ... 25
ISSN 1211 - 4286
Introduction
Early in the eighties it was shown that some newer com- plexing agents, e.g. sodium 2,3-dimercapto-1-propanesulp- honate (DMPS; unithiol), or meso-2,3-dimercaptosuccinic acid (DMSA), were effective in mercury, arsenic and lead poisoning. Compared to 2,3-dimercaptopropanol (BAL), the newer agents were of significantly lower toxicity.
Moreover, they can be administered orally or intravenously (1). Thus, it can be expected that older chelating agents (e.g.
BAL, CaNa2EDTA) will be replaced by DMPS and DMSA (2). Effectiveness of the first mentioned agent - DMPS - was also demonstrated in various experimental haevy metal in- toxications, e.g in As, Hg, Au intoxications (8,23,15).
In addition to haevy metal-chelating activity, DMPS as a dithiol agent may act as an oxygen radical scavenger and
thus it may inhibit lipid peroxidation (4,3). Oxidative stress plays an important role in various pathological conditions, e.g. in the anthracycline cardiotoxicity (20). In relation to the study of DMPS in the above-mentioned pathological state, we have studied changes of biochemical and haema- tological parameters in the course of repeated (10 weeks) intravenous DMPS administration in rabbits. In addition, myocardial content of some elements (calcium, potassium, magnesium, iron, selenium) was also measured at the ter- mination of the experiment.
Methods
Chemicals
2,3-dimercaptopropane-1-sulfonic acid, sodium salt (Sigma Chemie, Czech Republic); ketamin (Narkamon 5% inj., Léči-
3
ORIGINAL ARTICLE
EFFECTS OF REPEATED ADMINISTRATION OF DITHIOL CHELATING AGENT - SODIUM 2,3-DIMERCAPTO-1-PROPANESULPHONATE (DMPS) - ON BIOCHEMICAL AND HAEMATOLOGICAL PARAMETERS IN RABBITS
Vladimír Geršl1, Radomír Hrdina2, Jaroslava Vávrová3, Magdalena Holečková3, Vladimír Palička3, Jaroslava Voglová4, Yvona Mazurová5, Jiří Bajgar6
1Department of Pharmacology, Charles University, Faculty of Medicine, Hradec Králové;
(Head: prof. MUDr. J. Martínková, CSc.)
2Department of Pharmacology and Toxicology, Charles University, Faculty of Pharmacy, Hradec Králové;
(Head: prof. MUDr. Z. Fendrich, CSc.)
3Institute of Clinical Biochemistry and Diagnostics, University Teaching Hospital, Hradec Králové;
(Head: doc. MUDr. V. Palička, CSc.)
4Department of Clinical Hematology, University Teaching Hospital, Hradec Králové;
(Head: prof. MUDr. L. Chrobák, CSc.)
5Department of Histology, Charles University, Faculty of Medicine, Hradec Králové;
(Head: prof. MUDr. S. Němeček, DrSc.)
6Military Medical Academy, Hradec Králové;
(Head: doc. MUDr. S. Býma, CSc.)
Summary: The effects of weekly intravenously administered a dithiol chelating agent - sodium 2,3-dimercaptopropane- sulphonate (DMPS) - in a single dose of 50 mg/kg/week for 10 weeks on biochemical and haematological parameters were studied in rabbits. DMPS was well tolerated, an increase in body weight was similar in the DMPS-treated and control ani- mals. DMPS caused significant decrease in plasma calcium and vitamin E concentrations at the end of the experiment.
No significant differences in haematological parameters between the DMPS and control groups were observed. A signifi- cant decrease in magnesium content in myocardial tissue was observed in the DMPS-treated rabbits. The above-mentioned biochemical changes should be taken into account in studies of possible chelating and radical scavenging effects of DMPS in various pathological conditions.
Key words:2,3-dimercapto-1-propanesulphonate; DMPS; Chelating agents; Dithiols; Biochemistry; Haematology; Myocardial elements; Rabbit
ACTA MEDICA (Hradec Králové) 1997;40:3-8
va, Czech Republic); pentobarbital (Nembutal Sodium, Abbott, USA); aqua pro injectione (Biotika, Slovakia), sali- ne (Solutio natrii chlorati isotonica, Biotika, Slovakia).
Experimental design
Medium size Chinchilla male rabbits of average weight 3.0 kg and age 4 months at the beginning of the experiment were used. This experiment followed the Law of the Czech National Council for the protection of animals against cru- elty, as well as the European convention for the protection of vertebrate animals used for experimental and other sci- entific purposes of the Council of Europe, and was under the supervision of the Ethics Committee of the Medical Faculty, Charles University, Hradec Králové.
DMPS was administered i.v. to twelve rabbits in a dose of 50 mg/kg once weekly for 10 weeks. DMPS was dissol- ved in aqua pro injectione immediately before administrati- on in a concentration 50 mg/ml. Fifteen control rabbits were given saline i.v. in corresponding volume (1.0 ml/kg).
The weight of rabbits was monitored during the experi- ment. Biochemical and haematological parameters were de- termined in arterial blood samples (plasma, or serum, resp.) before the 1st and 5th administration of the drug and at the end of the experiment (3-5 days after the 10th admi- nistration of the drug). After the sacrifice of rabbits by i.v.
pentobarbital overdosing at the 11th week, the gross autop- sy was performed, heart was excised, sample of the left ven- tricle was removed for determination of elements content - calcium, potassium, magnesium, iron and selenium.
Biochemical parameters
The following biochemical parameters were determined in plasma (serum) samples using automatic analyser Hitachi 717 (Japan): Na, K, Ca, Cl, Mg, phosphate, gluco- se, urea, creatinine, uric acid, bilirubin, lactate dehydroge- nase (LD), alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatine kinase (CK), alkaline phosphatase (ALP), cholesterol, triglycerides, proteins incl.
electrophoresis, glutathione peroxidase (GSH-px), glutathi- one (GSH), malone dialdehyde (MDA), vitamin E.
Haematological parameters
Parallel to biochemical parameters, the following haematological parameters were determined using analyser Coulter T890 (USA): white blood cells count and white blood picture, red blood cells count, haemoglobin, haema- tocrit and thrombocytes count.
Myocardial content of elements
The content of calcium (Ca), potassium (K), magnesi- um (Mg), iron (Fe) and selenium (Se) was measured in samples of dry, mineralized (using microwave digestion with nitric acid and hydrogen peroxide in microwave oven, Milestone, Italy) left ventricular tissue and expressed in µmol/g (Se in nmol/g) of dry tissue. Ca, Mg and Se were determined by atomic absorption spectrophotometry using
an analyser Unicam Sollar 959 (USA). Content of K and Fe was measured photometrically using an apparatus Eppendorf Efox 5053 (Germany), and Hitachi 717 (Japan), resp.
Statistical analysis
The data are presented as means± S.E.M. Significance was estimated with the adequate t-test at the level p ≤ 0.05 (9).
Results
Body weight
The initial values of body weight were 3193± 68 g in the control group, and 3042± 60 g in the DMPS group, resp.
The weight gain in both groups of rabbits was almost iden- tical during the experiment (Fig. 1).
Fig. 1: Body weight gain (%) in the control and DMPS tre- ated rabbits during the experiment (11 weeks); p<0,005 compared to the initial value.
Biochemical parameters
As shown in tab. 1, there were some significant diffe- rences between control and DMPS treated animals at the beginning of the experiment in some measured biochemical parameters. In the DMPS group, there was a higher con- centration (or activity) of Na, urea, ALT, albumin, GSH-px and GSH, compared to the control group. No consistent trends were found in the measured parameters during the experiment (i.e., in the 5th week). At the end of the experi- ment, a significant decrease in Ca plasma concentration was found in DMPS treated rabbits compared to the con- trol group. In addition, there was a significantly lower inc- rease in CK activity in DMPS group vs.control group. In the middle of the experiment (the 5th week), there was also transient, but significant increase in some globulin fracti- ons (α2, β, γ) of plasma proteins in DMPS treated rabbits.
Compared to the initial values, there was a significant de- crease in vitamin E concentration at the end of the experi- ment in the DMPS group.
0 5
100 110 120 130
week
%
DMPS
7 8 9 10 11
*
*
*
*
*
*
*
*
*
*
*
*
control
Table 1:Biochemical parameters (initial = absolute values) and their changes (%) during repeated i.v. administration of DMPS (50 mg/kg/week).
Time interval (weeks) Parameter control group (n1=15) DMPS group(n2=12)
1 (initial) 5 11 (final)
sodium 141.7±0.7 101.2±0.7 101.9±0.8* (mmol/l) 144.5±0.7# 99.0±0.7# 99.8±1.2 potassium 4.0±0.2 93.6±3.2 83.9±5.0* (mmol/l) 3.9±0.1 92.4±4.1 106.0±11.6 chloride 102.2±1.0 102.4±1.2 100.6±1.5 (mmol/l) 104.3±0.8 97.7±1.0#* 103.2±2.7 calcium 3.1±0.1 102.7±4.6 96.5±2.2 (mmol/l) 3.2±0.1 106.4±2.2* 86.8±4.1#*
magnesium 0.94±0.08 359.3±253.0 893.6±792.8 (mmol/l) 1.02±0.03 96.5±8.7 99.9±4.0 phosphate 1.90±0.10 83.1±3.9* 77.2±4.7* (mmol/l) 1.85±0.07 85.8±3.8* 75.2±2.9* glucose 10.8±0.8 100.3±9.2 133.7±13.8* (mmol/l) 8.7±0.9 118.8±13.3 131.0±14.4 urea 7.9±0.3 111.7±6.3 101.4±4.8 (mmol/l) 9.7±0.3# 95.4±4.4 92.7±8.0 creatinine 91.2±4.1 106.9±2.8* 108.4±4.8 (µmol/l) 90.9±1.9 106.9±3.8 109.9±5.3 uric acid 12.5±1.8 72.3±11.6* 100.8±34.4 (µmol/l) 13.9±2.3 83.2±34.5 94.9±53.3 bilirubin 6.3±0.3 86.1±3.5* 84.0±3.4* (µmol/l) 6.3±0.2 81.0±2.4* 80.2±2.9* LD 7.1±2.0 249.1±80.3 300.3±139.0 (µkat/l) 7.8±1.0 121.8±26.5 108.8±19.7
ALT 1.2±0.1 102.1±6.6 92.2±7.4
(µkat/l) 1.8±0.1# 83.9±3.9#* 75.1±4.3* AST 0.49±0.05 146.7±16.8* 114.0±9.5 (µkat/l) 0.58±0.04 104.8±8.3# 118.2±15.7 CK 23.6±4.0 134.8±22.0 202.4±24.9* (µkat/l) 32.4±3.8 105.0±14.1 124.2±18.1# ALP 3.4±0.3 102.1±11.4 53.9±6.9* (µkat/l) 2.9±0.2 85.9±6.1* 55.7±5.3* cholesterol 1.04±0.11 119.9±11.4 79.4±8.3* (mmol/l) 1.15±0.09 94.2±5.6 214.6±145.4 triglycerides 1.03±0.09 91.7±8.0 133.1±13.0*
(mmol/l) 0.85±0.09 137.8±18.4# 168.7±19.4*
Table 1(cont’d)
protein (g/l) 62.3±1.1 107.4±2.7* 99.7±2.5 64.7±0.8 97.7±5.8 101.3±3.4
electrophoresis:
albumin (%) 66.3±1.3 103.1±1.1* 101.2±2.6 69.8±1.3# 95.1±2.0# 98.1±2.0 α1globulin 5.6±0.6 140.9±50.8 117.6±22.5
6.2±0.2 see note 96.3±8.6 α2globulin 6.7±0.4 92.3±5.3 97.6±7.0 5.2±0.4# 138.4±11.1#* 122.5±9.2#*
βglobulin 11.0±1.1 109.0±11.4 87.7±8.2 9.3±1.0 184.3±14.9#* 95.6±8.7 γglobulin 12.5±0.9 88.9±7.1 102.7±8.8 10.6±0.5 110.6±6.4# 111.0±7.5 A/G quotient 2.04±0.13 114.9±5.9 103.5±11.4
2.51±0.14# see note 90.6±6.1 GSH-px 84.0±7.8 216.9±22.7* 378.2±40.4* (U/g Hb) 124.2±9.9# 223.8±13.2* 376.4±26.7* GSH 5.4±1.2 511.1±222.3 414.2±176.1 (mg/g Hb) 9.6±1.2# 181.4±123.0 240.6±139.1 MDA 0.86±0.11 92.6±12.3 116.7±22.1 (µmol/l) 0.87±0.10 99.2±10.2 102.0±10.0 vitamin E 4.3±1.0 74.4±11.5 97.6±25.0 (µmol/l) 2.6±0.4 60.0±20.6 58.9±15.2*
LD - lactate dehydrogenase;
ALT - alanine aminotransferase;
AST - aspartate aminotransferase;
CK - creatine kinase;
ALP - alkaline phosphatase;
A/G quotient - albumin/globulin quotient;
GSH-px - glutathione peroxidase;
GSH - glutathione;
MDA - malondialdehyde.
Note: values not measured due to interference between fractions of albumin and α1globulin.
Statistical significance (p ≤ 0.05):
* - compared to the initial value;
# - between groups
Haematological parameters
Tab. 2 shows that there were no significant differences in haematological parameters between the DMPS and con- trol groups of rabbits at the beginning of the experiment.
Some changes observed during the experiment were com- parable in both groups.
5
Table 2: Haematological parameters (initial = absolute va- lues) and their changes (%) during repeated i.v. administra- tion of DMPS (50 mg/kg/week).
Time interval (week)
Parameter control group (n1=15)
DMPS group(n2=12)
1 (initial) 5 11 (final)
leucocytes 7.9±0.7 90.1±7.2 57.4±5.9* (109/l) 7.6±0.6 89.9±6.9 61.1±6.4* erythrocytes 5.8±0.2 109.4±2.1* 108.8±2.9* (1012/l) 5.9±0.1 110.3±2.7* 111.7±1.5* haemoglobin 120.9±3.0 113.2±1.9* 110.5±2.8* (g/l) 121.3±2.4 115.1±3.3* 117.4±2.2* haematocrit 0.377±0.081 112.1±2.3* 111.1±3.0* (ratio) 0.394±0.008 111.7±2.3* 112.6±2.1* MCV 65.7±0.5 102.6±1.1* 102.0±0.7* (fl) 66.4±0.5 101.2±1.1 100.6±0.9 thrombocytes 544.3±34.6 84.7±4.9* 83.7±3.9* (109/l) 595.3±42.2 75.9±5.3* 78.6±6.3*
differential count (%)
band 0.5±0.2 0.0±0.0 0.0±0.0
neutrophils 0.1±0.1 0.0±0.0 12.5±12.5* segment 41.5±3.5 97.4±12.1 104.5±18.2 neutrophils 40.3±4.8 98.6±16.6 125.1±19.5 eosinophils 2.5±1.0 19.7±12.1* 140.7±45.2 1.2±0.5 287.5±138.1 25.0±17.1#*
basophils 1.7±0.8 75.0±46.1 72.2±25.7 0.6±0.2 10.0±10.0* 66.7±66.7 monocytes 2.7±0.7 88.8±14.2 203.6±56.4 2.0±0.5 88.3±37.5 98.3±46.3 lymphocytes 48.5±3.8 134.4±11.8* 111.2±13.0 55.8±4.9 117.6±13.5 88.2±10.9
MCV- mean cellular volume.
Statistical significance (p ≤ 0.05):
* - compared to the initial value;
# - between groups
Myocardial content of elements
There was a tendency to a decrease in calcium and po- tassium content in the myocardium of DMPS treated rab- bits. In the case of magnesium, this decrase was significant.
The content of iron and selenium were not significantly dif- ferent in both groups of rabbits (tab. 3).
Table 3:The content of elements - calcium, potassium, mag- nesium, iron and selenium - in the left cardiac ventricle af- ter repeated i.v. administration of DMPS (50 mg/kg/week).
Group Ca K Mg Fe Se
control 17.7±3.1 436.9±69.7 65.1±6.8 9.6±1.8 10.0±1.2 (n1=15)
DMPS 11.2±1.3 390.5±29.5 47.6±4.6# 9.5±1.4 12.7±1.5 (n2=12)
Means± S.E.M. in µmol/g (Se-nmol/g) of dry tissue.
Statistical significance between groups: # p ≤ 0.05
Discussion
Repeated i.v. administration of DMPS in a cumulative dose of 0.5 g/kg/10 weeks was well tolerated by rabbits. The weight gain of the DMPS treated animals was comparable with that of the control group. After slow i.v. administrati- on of a single dose 50 mg/kg of DMPS only transient irre- gularities in respiration were observed. These results correspond to those of other authors who had studied chro- nic effects of DMPS in other species of experimental ani- mals (19, 22). Rabbits were used in our experiment considering a follow-up study of possible effect of DMPS on anthracycline cardiotoxicity, as the anthracycline model of chronic heart failure in rabbits has been often used for these purposes (24,5,10). Similarly, the selected single dose of DMPS (i.e. 50 mg/kg) corresponds on a molar basis to the doses of both experimentally and clinically used cardi- oprotective agent against anthracycline cardiomyopathy - dexrazoxane (ICRF-187) (12).
In some measured biochemical parameters (Na, urea, ALT, GSH-px, etc.) there were significant differences bet- ween the DMPS and control groups at the beginning of the experiment. This fact can be explained by the variation of the biochemical parameters in rabbits (17). A significant decrease in plasma calcium concentration in the DMPS treated rabbits may be possibly due to chelating activity of DMPS. The concentration of other ions, however, was not affected by DMPS treatment. An increase in CK activity in both groups of rabbits during the experiment can be expla- ined by repeated i.m. administration of an anaesthetic agent (ketamine). In the DMPS group this increase was signifi- cantly attenuated at the end of the experiment.
The cause of a significant decrease in vitamin E plasma concentration at the end of the experiment in the DMPS treated rabbits (compared to the initial value) may be com- plex. Rapid oxidation of DMPS after i.v. administration to disulfide forms (16,18) in the blood is supported by kinetic data. Fifteen minutes after i.v. administration of DMPS (3.0 mg/kg) to humans only 12% of the total DMPS was un- changed, i.e. nearly 90% of DMPS was oxidized to disulfi- des (14). In the cells disulfide forms of DMPS can be
reduced at the expense of vitamin E or by involving glutat- hione-disulfide exchange reaction (21). In addition, depleti- on of cytosol GSH may contribute to the attenuated regeneration of the membrane-bound vitamin E (7,13).
Doses of DMPS used in this study caused no significant changes in haematological parameters. Higher doses of DMPS can cause anaemia due to copper deprivation, as ob- served by Szinich et al. (22) in dogs treated i.v. with DMPS 2 x 75 mg/kg daily for 10 weeks.
In the DMPS group, there was a significant lower con- tent of Mg in the myocardium of the left ventricle. There was also tendency to a decrease in Ca and K myocardial content, in the case of Ca the decrease was near the level of statistical significance (p = 0.069). On the other hand, the content of Fe and Se was not practically affected by DMPS treatment. The described changes in mineral concentrati- ons, especially those concerned magnesium, calcium and partially iron, were consistent with findings of Bosque, et.
al. (1990).
In conclusion, experimental data of our study confir- med low toxicity of repeatedly i.v. administered DMPS in rabbits. In respect to a possible use of this dithiol chelating agent in other pathological states than haevy metal intoxi- cations (e.g. in those where an oxidative stress plays a role), one should take into account distinct effects of DMPS on the myocardial content of some important elements, parti- cularly of magnesium and calcium.
Acknowledgements
The author wish to thank Mrs. L. Koželuhová for her technical assistance, and Dr. E. Ettlerová for statistical eva- luation of the results. This study was supported by means of Internal Grant of the Charles University No. 256/1994, and by IGA MH CR No. 4212-3/97.
References
1. Aaseth J. Recent advance in the therapy of metal poiso- nings with chelating agents. Hum Toxicol 1983;2:257-72.
2. Aposhian HV, Maiorino RM, Gonzalez-Ramirez D, Zuniga-Charles M, Xu Z, Hurlbut KM, Junco-Munoz P, Dart RC, Aposhian MM. Mobilization of haevy metals by newer, therapeutically useful chelating agents. Toxicology 1995;97:23-38.
3. Benov LC, Benchev IC, Monovich OH. Thiol antidotes effect on lipid peroxidation in mercury-poisoned rats.
Chem Biol Interact 1990;76:321-32.
4. Benov LC, Ribarov SR, Monovich OH. Study of activa- ted oxygen production by some thiols using chemiluminis- cence. Gen Physiol Biophys 1992;11:195-202.
5. Bocherens-Gadient SA, Quast U, Nussberger J, Brunner HR, Hof RP. Chronic adriamycin treatment and its effect on the cardiac β-adrenergic system in the rabbit.
J Cardiovasc Pharmacol 1992;19:770-8.
6. Bosque MA, Domingo JL, Paternain JL, Llobet JM, Corbella J. Evaluation of the developmental toxicity of 2,3-
dimercapto-1-propanesulphonate (DMPS) in mice. Effect on mineral metabolism. Toxicology 1990;62:311-20 7. Coudray C, Boucher F, Deleiris J, Favier A. Modulation by selenium supplementation of lipid peroxidation induced by chronic administration of adriamycin in rats. Nutrition 1995;11 (Suppl.):512-6.
8. Flora SJ, Dube SN, Arora U, Kannan GM, Shukla MK, Malhotra PR. Therapeutic potential of meso- 2,3-dimercap- tosuccinic acid or 2,3-dimercaptopropane-1-sulphonate in chronic arsenic intoxication in rats. Biometals 1995;8:
111-6.
9. Gad SC, Weil Cs. Statistics for toxicologists. In: Hayes AW (ed). Principles and methods of toxicology. 2nd ed., New York:Raven Press, 1989;445-6.
10. Geršl V, Mazurová Y, Bajgar J, Mělka M, Hrdina R, Palička V. Lack of cardiotoxicity of a new antineoplastic agent, a synthetic derivative of indenoisochinoline: compa- rison with daunorubicin in rabbits. Arch Toxicol 1996;70:645-51
11. Geršl V, Hrdina R. Noninvasive polygraphic cardiac changes in daunorubicin-induced cardiomyopathy in rab- bits. Sb Ved Pr Lek Fak Univerzity Karlovy Hradec Králové 1994;37:49-55.
12. Green MD, Alderton P, Gross J, Muggia FM, Speyer JL. Evidence of the selective alteration of anthracycline ac- tivity due to modulation by ICRF-187 (ADR-529).
Pharmac Ther 1990;48:61-9.
13. Haenen GRMM. Thiols in oxidative stress. Some impli- cations for catecholamine toxicity. Amsterdam:Free University,1990.182 pp.
14. Hurlbut KM, Maiorino RM, Mayersohn M, Dart RC, Bruce DC, Aposhian HV. Determination and metabolism of dithiol chelating agents. XVI. Pharmacokinetics of 2,3- dimercapto-1-propanesulphonate after intravenous admi- nistration to human volunteers. J Pharmacol Exp Ther 1994;268:662-8.
15. Inns RH, Rice P. Efficacy of dimercapto chelating agents for the treatment of poisoning by percutaneously applied dichloro(2-chlorovinyl)arsine in rabbits. Hum Exp Toxicol 1993;12:241-6.
16. Klimmek R, Krettek C, Werner HW. Acute effects of the haevy metal antidotes DMPS and DMSA on circulati- on, respiration, and blood homeostasis in dogs. Arch Toxicol 1993;67:428-34.
17. Kraus AL, Weisbroth SH, Flatt RE, Brewer N. Biology and diseases of rabbits. In: Fox JG, Cohen BJ, Loew FM, eds. Laboratory animal medicine. Orlando:Academic Press,1984:212-3.
18. Maiorino RM, Weber GL, Aposhian HV.
Determination and metabolism of dithiol chelating agents.
III. Formation of oxidized metabolites of 2,3-dimercapto- propane-1-sulfonic acid in rabbits. Drug Metab Dispos 1988;16:455-63.
19. Planas-Bohne F, Gabard B, Schaffer EH. Toxicological studies on sodium 2,3-dimercaptopropane-1-sulphonate in the rat. Arzneimittel-Forsch 1980;30:1291-4.
7
20. Seifert CF, Thompson DF, Nesser ME. Dexrazoxane in the prevention of doxorubicin-induced cardiotoxicity. Ann Pharmacother 1994;28:1063-72.
21. Stewart JR, Diamond GL. In vivo renal tubular secreti- on and metabolism of the disulfide of 2,3-dimercaptopro- pane-1-sulphonate. Drug Metab Dispos 1988;16:189-95.
22. Szinicz L, Wiedemann P, Häring H, Weger N. Effects of repeated treatment with sodium 2,3-dimercaptopropane- 1-sulphonate in Beagle dogs. Arzneimittel-Forsch 1983;33:
818-21.
23. Takahashi Y, Funakoshi T, Shimada H, Kojima S. The utility of chelating agents as antidotes for nephrotoxicity of gold sodium thiomalate in adjuvant-arthritic rats.
Toxicology 1995;97:151-7.
24. Van Vleet JF, Ferrans VJ. Evaluation of vitamin E and selenium protection against chronic adriamycin toxicity in rabbits. Cancer Treat Rep 1980;64:315-7.
Submitted April 1997.
Accepted May 1997.
Doc. MUDr. Vladimír Geršl, CSc., Department of Pharmacology, Charles University, Faculty of Medicine, Šimkova 870, 500 01 Hradec Králové, Czech Republic.
Introduction
N-(3,5-dimethoxy-4-propoxyphenylethyl)-aziridine or FAZ-4 is the most interesting representative of groups of aziridine derivatives, which were formerly synthetised at the Chemical centre of the Slovak Academy of Sciences in Bratislava according to the original US patent (13).
We estimated the toxicity of aziridine derivatives follo- wing the method of Weil (16). These compounds are mild- ly toxic, but FAZ-4 was the most toxic within them (7). We also studied the effects of intracerebral administration of FAZ-4 on some qualitative indicators of learning and me- mory, FAZ-4 induced a disturbance of spatial orientation in a T maze. The influencing of operant conditioning in the Skinner box had a very similar course of events (8,9).
Furthermore, FAZ-4 was the only aziridine derivative tes- ted which possesses the convulsive activity. This one is comparable with pentylenetetrazol (PTZ) paroxysm (see, e.g.4,11),with one exception: the lack of tonic component of major PTZ paroxysm (5). Presuming different sites of origin for the tonic and clonic component, i.e., the forebra- in in case of clonic, and the brainstem in case of tonic com- ponent (3,14,15), we can therefore exclude the brainstem as the site of convulsive activity of FAZ-4. The aim of the pre- sent work is an evaluation of two anticonvulsants with dif- ferent mechanisms of action, i.e. alprazolam and ketamine, with respect to FAZ-4 induced seizure activity. This contin- gent anticonvulsive efficacy of the both drugs tested is also compared with the same ability of them in the case of clas- sical convulsive agent PTZ.
Material and Methods
The experiments were carried out on a total number of 144 male Wistar rats (VELAZ, Praha), weighing 200 - 260 g.
The animals were fed with standard Larsen diet and had free access to water. The rats were randomly divided into groups of 8 animals. Occurence of abnormal signs, especi- ally the feature and intensity of the convulsions, was evalu- ated according to specially elaborated scale (10,11), described in details previously (5). Maximum rate in this scale (equal to 5) corresponds to the generalized tonic-clo- nic seizures (convulsions) represented by the loss of the righting reflex at the beginning of the tonic phase and tonic- clonic seizures involving muscles of forelimbs, hindlimbs, and whole body.
FAZ-4 was administered intraperitoneally in a dose of 50 mg/kg. Its effect was compared with the model con- vulsive drug PTZ, injected subcutaneously in a dose of 100 mg/kg.
Both convulsants were freshly dissolved in saline.
Alprazolam (VÚFB, Prague) and ketamine (KetalarR, Parke- Davis, Madrid) were given intraperitoneally either 30 minu- tes prior, or 1 minute subsequently, to administration of the convulsants tested. Doses used were 0,1 and 0,5 mg/kg for alprazolam, 10 and 25 mg/kg for ketamine. Following the administration of the drugs, the animals were placed into the experimental cage and individually observed for a peri- od of 30 minutes after the last injection. The animals were always assigned the highest convulsive score observed and the average score was calculated for all groups (always 8 animals in each) and doses. The lethality of the animals wit- hin 48 hours after all experiments was recorded. The results were statistically evaluated by means of t-test. The level of significance was set at the 5% level.
Results
A lack of any tonic component of major paroxysm fol- lowing DSP-4 was confirmed with respect to PTZ (compa-
9
ORIGINAL ARTICLE
EFFECT OF ALPRAZOLAM AND KETAMINE ON SEIZURES INDUCED BY TWO DIFFERENT CONVULSANTS
Josef Herink
Military Medical Academy J. E. Purkyně, Hradec Králové;
(Rector: doc. MUDr. S. Býma, CSc.)
Summary: Effect of two anticonvulsants with different mechanism of action, i.e. alprazolam and ketamine, was tested in two types of seizure activity. The first one was induced by N-(3,5-dimethoxy-4-propoxyphenylethyl)-aziridine, the second one by pentylenetetrazol. While alprazolam alleviated both the minimal as well as major paroxysms, ketamine supressed only ma- jor seizures. These effects are discussed in terms of the both N-methyl-D-aspartate and GABA receptors involvement.
Key words:Aziridines; Convulsions; Pentylenetetrazol; Rat; Experimental Therapy
ACTA MEDICA (Hradec Králové) 1997;40:9-11
re upper and bottom left columns in Figures). Doses of both convulsants tested represent more than LD50 value.
Alprazolam exerted a biphasic effect on the PTZ con- vulsions. The lower dose used increased intensity of these symptoms, while for the higher dose a strong anticonvulsi- ve effect, especially in case of subsequent administration, was observed (Fig. 1, above). On the contrary, alprazolam always showed anticonvulsive effect against FAZ-4 induced seizures, with exception of prior administration of the lower dose used (Fig. 1, below). In addition to it, alprazolam had a strong positive effect on sur vival of animals following ad- ministration of all doses of convulsants tested.
Fig. 1:The influence of alprazolam (ALP) on PTZ-induced (top) and FAZ-4-induced (bottom) seizure activity. The re- sults are represented as means with SD of a total score of seizure activity. The numbers (0.1, 0.5) indicate used doses of ALP. The letters represent the order of drugs tested ad- ministration (i.e. prior or subsequent administration of ALP with respect to the convulsants tested). The numbers in the base of each column indicate the lethality (number of died animals to the total number of animals in each group).
Ketamine suppressed the PTZ convulsions at the higher dose tested (Fig. 2, above), subsequent administration of this drug (i.e., 1 minute after PTZ) was more effective than that of prior injection (i.e., 30 minutes before PTZ).
Ketamine was more effective in the suppression of the FAZ-4 induced convulsion in case of subsequent administ- ration of both doses tested (Fig. 2, below). Ketamine de- creased significantly the lethality in animals. This protection was more evident in PTZ than in DSP-4 intoxi- cation.
Fig. 2: The inf luence of ketamine (KTM) on PTZ-induced (top) and FAZ-induced (bottom) seizure activity. Other de- tails as in Fig. 1.
Discussion
Convulsive effects and their influence is connected di- rectly or indirectly with number of neuromediators, similari- ty as it was demonstrated for cholinergic and peptidergic sys- tems (1,4) or organophosphates (2). PTZ is the prototype agent in the class of systemic convulsants. Nevertheless its
PTZ 1
2 3 4 5 SCORE
ALP/PTZ 0,1
PTZ/ALP 0,1
ALP/PTZ 0,5
PTZ/ALP 0,5
FAZ 1
2 3 4 5 SCORE
ALP/FAZ 0,1
FAZ/ALP 0,1
ALP/FAZ 0,5
FAZ/ALP 0,5
PTZ 1
2 3 4 5 SCORE
KTM/PTZ 10
PTZ/KTM 10
KTM/PTZ 25
PTZ/KTM 25
FAZ 1
2 3 4 5 SCORE
KTM/FAZ 10
FAZ/KTM 10
KTM/FAZ 25
FAZ/KTM 25
mechanism of action is only poorly understood. At a syna- ptic level PTZ appears to interact with the GABA receptor- benzodiazepine complex (4,12). However other mechanisms for PTZ-induced seizures must also be important, we suppo- se that it would be NMDA-receptors involvement with res- pect especially to the major paroxysms.
Anticonvulsant tested suppressed seizures in different manner. Alprazolam was effective against minimal as well as major seizures, while ketamine suppressed only major seizures. Regarding to presumed N-methyl-D-aspartate (NMDA) antagonistic properties of ketamine, there is a litt- le possibility of the involvement of these receptor system in the origin and propagation of minimal seizure phenome- non (14,15). Strong anticonvulsive activity of benzodiazepi- nes, especially alprazolam, suggests the GABA receptor- chloride ionophore complex involvement in the origin and propagation not only in case of PTZ- but FAZ-4 convulsi- ons too (6,7). Discrepancy between „proconvulsive“ effect of lower dose tested of alprazolam on the one hand, and de- creased lethality of animals in the same situation on the se- cond hand, excludes largely straightforward chain of events leading from major paroxysm to the death itself.
Acknowledgement:
Author wish to thank Mrs. J. Bajgarová for skilfull technical assistance.
This work was supported by grant No. 307/95/1537 from the Grant Agency of the Czech Republic.
References
1. Bajgar J. Biological monitoring of exposure to nerve agents. Br J Ind Med 1992;19:648-53.
2. Bajgar J, Herink J, Skopec F, Hrdina V, Patočka J, Vičar J. Modulation of cholinergic nerve transmission by leucin5- enkephalin. Sb Ved Pr Lek Fak Univeryity Karlovy Hradec Králové 1985;28:47-54.
3. Browning RA. Role of the brain-stem reticular formation in the tonic-clonic seizures: Lesions and pharmacological studies. Fed Proc 1985;44:2425-31.
4. Fisher RS. Animal models of the epilepsies. Brain Res Rev 1989;14:245-78.
5. Herink J. Convulsive properties of N-(3, 5-dimethoxy-4- propoxyphenylethyl)-aziridine and their influencing by dia- zepam and triazolam. Sb Ved Pr Lek Fak Univerzity Karlovy Hradec Králové 1995;38:85-8.
6. Herink J, Koupilová M, Bajgar J. Effect of triazolam in convulsions induced by pentylenetetrazol. Homeostasis 1991;33:181.
7. Herink J, Koupilová M, Krs O, Bajgar J, Patočka J.
Modelling of some neuropathological states of the central nervous system by aziridine derivatives. Cesk Fyziol, 1992;41(suppl.):7-10.
8. Koupilová M, Herink J. Attend to antagonize DSP-4 in- duced impairment of the performance of rats in a T-maze.
Homeostasis 1994;36:41-3.
9. Koupilová M, Herink J, Bajgar J. Effects of aziridine de- rivative N-(3,5-dimethoxy-4-propoxyphenylethyl)-aziridine on learning and memory in laboratory rats. Homeostasis 1993;34:117-20.
10. Mareš P, Marešová D, Schickerová R. Effect of antiepi- leptic drugs on metrazol convulsions during ontogenesis in the rat. Physiol Bohemoslov 1981;30:113-21.
11. Mareš P, Mirvaldová H, Bělská M. Influence of a new antiepileptic drug ORG637O on metrazol-induced seizures in rats during ontogenesis. Physiol Bohemoslov 1990;39:
199-205.
12. Olsen RW. The GABA postsynaptic membrane recep- tor-ionophore complex. Site of action of convulsant and an- ticonvulsant drugs. Moll Cell Biochem 1981;39:261-79.
13. Razdan K. US patent office 3.637.622, 1972.
14. Velíšek L, Kusá R, Kulovaná M, Mareš P. Excitatory amino acid antagonists and pentylenetetrazol-induced sei- zures during ontogenesis. I. The effects of 2-amino-7-phosp- hoheptanoate. Life Sci 1990;46:1349-57.
15. Velíšek L, Kulhánková I, Roztočilová L, Mareš P.
Ethosuximide affects both pentylenetetrazole-and kainate- induced clonic seizures. Can J Physiol Pharmacol 1989;67:1357-61.
16. Weil CS. Tables for convenient calculation of median-ef- fective dose (LD50 or ED50) and instruction in their use.
Biometrics 1952;8:249-63.
Submitted May 1997.
Accepted June 1997.
MUDr. Josef Herink, DrSc., Purkyně Military Academy, P.O.Box 35, 500 01 Hradec Králové, Czech Republic.
11
Introduction
Anal fissure remains one of the most common procto- logic problems. Fissures are being seen in all age groups, although the majority of patients is represented by relative- ly young to middle-aged adults. Fissures cause considerable pain, irrelevant to the size of the lesion, resulting in signifi- cant morbidity and disability. Therefore,it is worthwhile to find and identify the most efffective but safe treatments for this disease.
A number of reports (6,8,9,10,12) have advocated the use of partial internal sphincterotomy as a treatment of chronic anal fissure. Surgical sphincterotomy promises the immediate relief of pain, it is simple to be performed and most patients express their satisfaction with the result.
However, the complications of sphincterotomy generally may be represented by a disquieting incidence of postope- rative incontinence to feces or flatus.
This study shows the result of a retrospective analysis of our patients who underwent lateral internal sphincterotomy for the treatment of chronic anal fissure.
Patients and methods
From January 1985 to December 1996, 133 patients (81 males, 52 females; average age 45,24 years) with chronic
symptomatic anal fissures underwent surgical treatment using internal lateral sphincterotomy.
As chronic fissures we considered those (according to Tzu-Chi Hsu), in which at least two of following three symptoms - sentinel pile, fissure with sclerotic edges with revealed interior sphincter on the bottom and hypertrophic anal papilla - were present.
Most of the operations were performed by the author.
Under general anesthesia in gynecological position the in- vestigation of anal canal was done first. After careful dif- ferentiation of the internal sphincter the right sided internal sphincterotomy in necessary extent was perfor- med while the anal canal tonus was controlled with left fo- ref inger. We were not bound to performe internal sphincterotomy in any patient as far as to the area of linea dentata.After hemostasis by help of compression we per- formed the suture of the incision by one stitch. Sentinel pile and hypertrophic anal papilla were cut only in cases of their larger size.
Post operation treatment was simple, occasionally anal- getics were given but dietetical restrictions were not neces- sary. In the anal hygiene we recommended short time sitting baths. After short hospitalization all patients were followed up in the outpatients department till total recovery.
To determine long time results we examined random sample of 75 (56%) operated patients. From these indivi-
13
ORIGINAL ARTICLE
THE CHRONIC ANAL FISSURE TREATMENT BY INTERNAL PARTIAL LATERAL SPHINCTEROTOMY AND FOLLOWING ANAL MANOMETRY
Dušan Šimkovič, Milan Široký, Karel Šmejkal
Department of Surgery, University Teaching Hospital, Hradec Králové;
(Head: doc. MUDr. J. Bedrna, CSc.)
Clinical Centre, Department of Biochemistry and Diagnostics, University Teaching Hospital, Hradec Králové;
(Head: doc. MUDr. J. Bureš, CSc.)
Summary: Anal fissure remains one of the most common proctologic problems. A number of reports have advocated the use of partial internal sphincterotomy as a treatment of chronic anal fissure. Our study shows the results of a retrospecti- ve analysis of our patients who underwent lateral internal sphincterotomy for the treatment of chronic anal fissure.
To determine long time results we examined random sample of 75 operated patients. Apart from taking careful history and patient’s assessment of the operation effect, the patients were investigated „per rectum“, and even possibly rectoscopical- ly. To asscertain suitability and also security of properly done internal sphincterotomy, we performed anorectal manome- tric examinations in 53 patients controlled. To conclude our results, we can state that in 9 (12%) patients controlled some subjective complaints or certain pathologic findings connected with anal fissure or sphincterotomy were found.
None of the people from the set of ours suffered from any complaints, which can be taken as stool incontinency. In no pa- tients any change in pressure measured by anorectal manometry indicating incontinency was proved.
Key words: Chronic anal fissure; Internal lateral sphincterotomy
ACTA MEDICA (Hradec Králové) 1997;40:13-15
duals there were 43 males, 32 females, average age, 45,12 years. Apart from taking careful history and patient’s as- sessment of the operation effect, the patients were investi- gated „per rectum“, and even possibly rectoscopically.
To asscertain suitability and also security of properly done internal sphincterotomy we performed anorectal ma- nometric examinations in 53 patients controlled.
Results
Having evaluated results of check ups we found that 70 controlled patients (93%) take the operation results for very satisfactory or good. When we investigated these individu- als digitally „per rectum“ we found no pathological change, which could have been in possible connection with anal fis- sure or sphincterotomy. Two patients from the set of ours complained of mild soiling, 2 other people occasionally had flatus control problems. In the two last mentioned ones we found manometric resting preassures at the lower limits wit- hin the normal range (40 mm Hg).
One patient of ours took the operation results for being satisfactory. When investigated the patient had a submu- cous perianal fistule connected with sphincterotomy per- formance.
Only 4 patients considered the operation results as un- satisfactory ones. One of them had perianal fistule in the spot of sphincterotomy, 3 others had signs of fissure recur- rency. Manometric measurement in two of those with fis- sure recurrences proved high levels of resting and squeeze pressures.
To conclude, we can state that in 9 (12%) patients con- trolled some subjective complaints or certain pathologic findings connected with anal fissure or sphincterotomy were found.
None of the people from the set of ours suffered from any such complaints, which can be taken as stool inconti- nency. In no patient any change in pressure measured by anorectal manometry indicating incontinency was proved.
Discussion
Anal fissure remains one of the most commonly en- countered proctologic problems. It affects both men and women of all ages. Its appearance is that of a relatively mi- nor lesion, but anal fissure causes considerable pain and discomfort, by far disproportionate to its size. Fissures are most commonly found in the posterior midline.
Identification of etiologic, as well as predisposing factors, may help to reduce the incidence of anal fissures.
Much work has been done to elucidate its etiology;ho- wever, a simple unification has not been established (1,10).
Anal sphincteric measurements show that the maximal basal pressure is significantly higher in patients with anal fissures and when these patients are treated by sphinctero- tomy, the basal pressure is significantly reduced. These stu-
dies suggest that increased pressure in the anal canal con- tributes to the pathophysiology of anal fissure (2,4).
Till the middle of this century, posterior sphincteroto- my was an accepted surgical treatment for chronic anal fis- sure (3). In 1969, Notaras (9) described a simpler and more effective method of partial lateral internal sphincte- rotomy. The advantage of this method over the posterior technique or stretching method was confirmed by Hawley (5) and Hoffman and Goligher (6).
Why some patients after partial sphincterotomy have transient or permanent postoperative incontinence of fla- tus or feces and others have not, is not clear, but the reason may be elucidated by the wide range of postoperative sp- hincter pressures. The lowest postoperative sphincter pres- sures reported are within the range usually associated with incontinence. These low pressures may result from sphin- cterotomy being carried out in patients with a fissure but with normal preoperative pressures (7).
Postoperative incontinence of flatus or feces, even if only mild soiling, may be extremely inconvenient. The conclusion reached by Pernikoff et al (11) in 1994 study of 500 sphincterotomies is representative to most of series. In spite of a long-term complication rate of 11% these authors state: „Partial lateral sphincterotomy is a highly succesful, safe procedure when performed carefully, selectively and under controlled circumstances“.
Surgical sphincterotomy is an excellent procedure for the long-term cure of chronic anal fissure. It probably works by reducing resting anal pressure by as much as 50%.
There is an evidence from laser Doppler studies that this may lead to improved perfusion of the pecten which might then facilitate healing of what are now known as relatively ischemic ulcers (2,7).
Conclusion
Careful patient selection, absence of peroperative con- tinence problems, and meticulous surgical techniques are necessary to achieve the type of results presented.
References
1. Arnous J, Denis J. Pathogenesis and concepts of anal fis- sure. Am J Proctol 1971;22:184-6.
2. Boulos PB, Araujo JGC. Adequate internal sphinctero- tomy for chronic anal fissure: subcutaneous or open tech- nique? Br J Surg 1984;71:360.
3. Eisenhammer S. The surgical correction of chronic anal (sphinteric) contracture. S Afr Med J 1951;25:486-9.
4. Hancock BD. The internal sphincter and anal fissure. Br J Surg 1977;64:92.
5 Hawley PR. The treatment of chronic fissure-in-ano: a tri- al of methods. Br J Surg 1969;56:915-8.
6. Hoffman DC, Goligher JC. Lateral subcutaneous inter- nal sphincterotomy in treatment of anal fissure. Br Med J 1970;3:673-5.
7. Chowcat NL, Araujo JG, Boulos PB. Internal sphincte- rotomy for chronic anal fissure: long-term effects on anal pressure. Br J Surg 1986;73:915-6.
8. Millar DM. Subcutaneous lateral internal anal sphincte- rotomy for anal fissure Br J Surg 1971;58:737-9.
9. Notaras MJ. Lateral subcutaneous sphincterotomy for anal fissure: A new technique. Proc R Soc Med 1969;62:713.
10. Oh C. Lateral subcutaneous internal sphincterotomy for anal fissure. Mt Sinai J Med (NY) 1975;42:596-601.
11. Pernikoff BJ,Eisenstat TE, Rubin RJ, Oliver GC, Salvati EP. Reappraisal of partial lateral internal sphincterotomy.
Dis Colon Rectum 1994;37:1291-5.
12. Rudd WW. Lateral subcutaneous internal sphincteroto- my for anal fissure. Dis Colon Rectum 1975;18:319-23.
Submitted May 1997.
Accepted May 1997.
MUDr. Dušan Šimkovič, CSc., Čechova 759, 500 02 Hradec Králové, Czech Republic.
15
Introduction
Profiling urinary organic acids is considered an invalu- able tool for diagnosing the numerous metabolic diseases known as organic acidurias (2-5, 8). These are a group of genetic disorders in which an enzyme or cofactor defect in one of the metabolic pathways leads to accumulation and increased urinary excretion of specific acidic metabolites.
Urinary organic acids (carboxylic acids and their amino acid conjugates) are water-soluble end products or interme- diates of the metabolism of amino acids, sugars, lipids, ste- roids, biogenic amines and many other compounds. It should be pointed out that the products themselves are nor- mal intermediates of the pathways involved, and it is the ab- normal amounts present which is pathologic. Accumulated organic acids may be found in blood (organic acidemias), but in much lower concentration compared to urine (orga- nic acidurias).
Organic acidurias manifest with an acute onset either early after birth or whenever later in life, often after provo- cation with a banal respiratory infect, vaccination or star- vation. Otherwise, progression of neurological symptoms may be the only sign of disease.
Symptoms of the organic acidurias include vomiting, dehydration, tachypnoe, abnormal odour, seizures, hypoto- nia, hypertonia, ataxia, hepatomegalia, lethargy and death.
Chronic manifestations are failure-to-thrive and mental re- tardation. Biochemical abnormalities, such as acidosis, hy-
poglycemia and hyperammonemia are due to increased pro- duction of ketoacids and to the accumulated organic acid intermediates, which then inhibit other pathways.
Prompt diagnosis of the specific enzyme abnormality and early treatment may prevent life threatening episodes and psychomotor retardation in some disorders.
The variety of clinical features and the similarity of symptoms for completely different disorders increase the importance of identifying excreted organic acids, direct as- say for deficient enzyme usually being either time-consu- ming or not widely available.
Recently we have introduced a three-step procedure for detection of organic acid abnormalities, the first using TLC methodology, the second GC and the third GC-MS techni- que.
Experimental
Materials
Organic acid standards were supplied by Sigma-Aldrich and Serva, silylating agents (BSTFA:TMCS) were from Pierce, precoated micropulverised cellulose thin-layers (plastic sheets No. 5577) and all other chemicals were pur- chased from Merck.
A fasting plasma and fresh random or 24-hrs-urine sam- ples should preferably be taken before the therapy is intro- duced. Cerebrospinal fluid is analysed in neurometabolic diseases and vitreous humour in post-mortem diagnoses.
17
BRIEF COMMUNICATION
SCREENING FOR ORGANIC ACID DISORDERS
Eliška Marklová, Miluše Brátová* and Viktor Voříšek**
Department of Pediatrics, Faculty Hospital, Charles University, Hradec Králové;
(Head: doc. MUDr. E. Pařízková, CSc.)
*Department of Gerontology & Metabolism, Faculty Hospital, Charles University, Hradec Králové;
(Head: prof. MUDr. Z. Zadák, CSc.)
** Department of Clinical Biochemistry & Diagnostics, Faculty Hospital, Charles University, Hradec Králové;
(Head: doc. MUDr. V. Palička, CSc.)
Summary: The detection of organic acidurias is a part of our screening programme for inherited metabolic diseases.
Adapted procedure is differentiated and involves several steps: 1) thin-layer chromatography (TLC) in the case of an ab- normal finding followed by 2) gas chromatography (GC). The next step of the investigation, using 3) gas chromatography mass-spectrometry (GS-MS) is reserved for more complicated and dubious analyses. In acutely sick patients and in the case of discrepancies between TLC results on the one hand, and clinical symptoms, supported by other laboratory findings on the other, the GC or GC-MS-analysis is performed immediately. Some examples of metabolic disorders, identified by this procedure, are presented.
Key words:Organic acids; Screening; Inherited metabolic disorders
ACTA MEDICA (Hradec Králové) 1997;40:17-19
