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A study conducted by J. Dubois, J. Colard, and H. L. Vis in 1966, focusing on muscle electrolyte composition and determination by neutron activation. The researchers used the BR1 and BR2 reactors of the C.E.N. in Mol, Belgium, to analyze the weights and electrolyte content of muscle biopsies taken from infants. The document also discusses the analytical methods used, including weighing, fat extraction, and gamma spectrometry.
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JOURNAL OF NUCLEAR MEDICINE 7:827-836, 1966
J. Dubois,2 J. Colard,3 and H. L. Vis
Brussels, Belgium
INTRODUCTION Because muscle tissue is abundant, easily approachable and of relatively constant composition, both hydrosaline and energetic metabolism can be easily studied in the muscle area. The activation analysis method, recently applied by Bergstrom (1) to the study of this material allows electrolytes (Na, K, Cl) and phosphor determina tions, on small muscular fragments taken by needle biopsies. This method has been adapted to the irradiation characteristics of the BR1 and BR2 reactors of the C.E.N. Mol (Belgium). In order to increase the speed and precision of the determination of induced activity, a new technique of measurements was devised. The purpose of this work, of which preliminary results are described in this paper, is a direct observation of the muscular hydrosaline equilibrium in some states of infantile dehydration.
MATERIAL AND METHOD
described by Bergstrom (1), who modified the Pollen and Bickel needle, which is pointed and hollow with a window at its distal end. A cylinder with a sharp rim fits into the needle and permits the sampling of material engaged in the window. Finally, a stylet may be introduced in the cylinder in order to extract the sample (Fig. I).
‘This work was done under contract Euratom-Université de Bruxelles-Université de Pise 026-63-4 Biac. 2Service et Laboratoire de Pédiatrie (Prof. R. DUBOIS) Université Libre de Bruxelles (Belgium). 3Centre d'Etude de l'Energie Nucléaire(Mol)—Belgium. 827
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II 4
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12
13 Fig. la. Needle1 used for muscle biopsy similar to that used by Bergstrom (1) but modified for pediatric investigation.
‘Manufactured in Belgium b@ SIMAL (CEMBLOUX).
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Fig. lb. From right to left: 1) Hollow needle window diameter (0.6 cm). 2) Cylinder with a sharp rim (diameter 0.2 cm). 3) Stylet (diameter 0.15 cm).
Half lifeCapture
cross section act.)-yf3Na2415.0 (oP.
h0.52 barn1. 100%1.39\lev100%K4212.5 2.75MevMev100%
h1.17 (^) barn1.53Mev18%3. 18%Cl'8 1.98\levMev82%
p3237.
mm.
143 days0.
barn
0.21 barn1.
2.15Mev\Iev
—31%
1.71Mev
Mev Mev @\Iev31%
MUSCLE ELECTROLYTE COMPOSITION AND NEUTRON ACTIVATION 831
The isotopes produced are Cl38 with a half-life of 37.5 mm. emitting 1. Mev and 2.15 Mev gamma rays and Na24 with a half-life of 15 h. emitting 1. and 2.75 Mev gamma rays (Table I). A sample of 1 ml of a solution of NaCl of known concentration, (O.OIM), is irradiated simultaneously with each group of biopsies. After irradiation, this
(Intertechnique) for five minutes and automatically recorded on a magnetic tape. The following sample is measured and recorded immediately afterwards. A perfect stability of the spectrum is obtained by means of a drift stabilizer, “Stabimat―,centered on the Na24 2.75 MeV photopeak.
A reference spectrum of Na24 is recorded on a magnetic tape in the same conditions as those of the samples. To obtain the relative activities, this spectrum
ISOTOPE OF SoDIuM, POTASSIUM, CHLORINE AND PHOSPHORUS OBTAINED BY NEUTRON ACTIVATION.
MUSCLE ELECTROLYTE COMPOSITION AND NEUTRON ACTIVATION 833
containing a carrier of KH2PO4 102M. Three samplings of the solution are pipetted off on counting trays and dried under an infrared lamp.
photomultiplier connected to a monochannel analysor.
tection of P32 ( max. : 1.7 MeV ) the monochannel analysor window width is 2
cause of the presence of sodium-24. In reality, the gamma rays of 2.75 MeV
detection of Na24 $ ray (1.39 MeV) and of the Compton event of one of the y radiations of Na24 can give an impulse exceeding the 2 MeV threshold fixed at the monochannel. In the material studied, this error can reach 25% of the total activity. The measurement of @32is done on the same counting trays after 10 days, allowing time for the decay of K42 and Na24. In this case, the monochannel window set from 0,4 to 1,8 MeV does not de tect the S35 and Ca45. The K and P contents of the muscular sample are calculated in reference to the standards.
PRELIMINARY RESULTS AND DISCUSSION Preliminary results are shown in Table II and III. Table II points out the values obtained by chemical and neutron activation methods in normal children,
by the two methods. We emphasize the fact that all our biopsies were done on children, because the values are different from those obtained in adults and cannot be compared. Similarly, Bergstrom's values for normal adults by the neutron activation method are identical with those found by other authors (3,4,5,6,7,8) by chemical
are comparable. Table III shows quantities of total water and electrolytes found in children with two different acute dehydration states (Isotonic and hypertonic dehydra tions). We have classified patients as hypertonic if serum sodium concentrations are 150 mEq/liter or higher. The preliminary results seem to indicate a rapid participation by muscular tissue in water and electrolyte movements which are characteristics of the above states. Moreover, these variations are generally similar to those obtained in the plasma. This seems to show that the muscular water osmolarity is well depicted by the plasmatic ionogram. In certain conditions, however, the interpretation of the data is ambiguous.
we assume that the dry tissue remains constant during all pathological processes. This assumption is probably true in acute states of dehydration occuring in pre viously healthy children, but we have noted that this material was modified in some cases of chronic malnutrition (7). In these last cases, the extracellular protein material composition is quan titatively different from that of normal muscle; the collagenous fractions become greatly augmented in comparison with the other extracellular protein fractions. Moreover the relation of total extracellular proteins to intracellular proteins changes, that is, referring the amounts of electrolytes to dry fat free weight will lead to a false interpretation. For example, the potassium values will be over estimated. We have to admit, therefore, that reference to fat-free solids will be inade quate in some circumstances. As do Lilienthal and his colleagues (9), some authors suggest expressing the results in terms of non-collagen nitrogen, that is, in terms of intracellular pro teins and we have indeed demonstrated that non-collagen nitrogen remains con stant even in severe pathological cases where collagen nitrogen is modified (10). It is impossible, however, to determine non-collagen nitrogen accurately on small fragments taken by needle biopsy. A surgical biopsy is necessary, but cannot be carried out in most cases.
i\ltSCt'LAR COMPOSITION IN TOTAL WATER, ELECTROLYTES AND PHOSPHORUS IN NORMAL CHILDREN, MEAN AND S.D.' Column A: results obtained by neutron activation. Column B and C: results obtained in two groups of children by chemical methods (7).
Total water 354.7 ± 6.2 342.4 ± 5.0 346.2 ± 5. —9 cases Na 18.4± 1.7 25.8± 1.0 16.8± 1. — 10 cases
—9cases (1 13.5± 1.5 15.4± 1.1 17.4± 0. — 10 cases 26.6± 1.4 29.9± 1. —8 cases —6 cases —10 cases
‘Theresults are expressed in g. for water and in mM for electrolytes and phosphorus, per 100 g. dry fat free solid.
SUMMARY AND CONCLUSIONS Following the work of Bergstrom in the adult, we have described a method of analysis of muscular electrolytes and phosphorus in children. By comparing our results with those obtained by the chemical technique we came to the con clusion that this method is reliable. An analysis of needle biopsy specimens of muscle from children with differ ent degrees of dehydration suggests that the muscular tissue is rapidly involved in acute hydro-electrolytic disorders. The preliminary data will be checked by the study of a larger series of cases. Although the use of such a method is limited, it remains the only one which allows a direct study of tissue in pediatric pathology.
REFERENCES