1. Fasting and Skeletal Muscle Enzymes in Obese Men Horm.
metabol. Res. 15 (1983) 271 Rath, R., K. Vondra: Short-term fasting
in the treatment of obesity. Vondra, K., R. Rath: Obesity and
thyroid function. 2. The effect Nahrung 21: 193-197 (1977) of
prolonged caloric restriction on Achilles tendon reflex values.
Rath, R., Z. Slabochova, K. Vondra: Immunoreactive insulin in
Endokrinologie 66: 332-336 (1975) obesity of adult women. Intemat
J. of Obesity 1: 279-286 Vondra, K., R. Rath, A. Bass: Skeletal
muscle HK activity and (1977) fasting FFA blood level in man. Horm.
Metab. Res. 8: 323 (1976) Vondra, K., R. Rath: Obesity and thyroid
function. 1. Values of Vondra, K., R. Rath, A. Bass, L. Kukla, Z.
Slabochova: Effect of the Achilles tendon reflex. Endokrinologie
62: 310-320 (1973) protracted intermittent fasting on the
activities of enzymes in- Vondra, K., R. Rath, Z. Kroupa: Improved
needle for muscle volved in energy metabolism, and on the
concentration of glyco- biopsy. Klin. Wschr. 52: 747-748 (1974)
gen, protein a DNA in skeletal muscle of obese women. Nutr. Vondra,
K., R. Rath, A. Bass: Activity of some enzymes of energy Metab. 20:
329-337 (1976) metabolism in striated muscle of obese subjects with
respect Vondra, K., A. Bass, V. Brodan, E. Kuhn, M. Andel, A.
Veselkova, to body composition. Horm. Metab. Res. 7: 475-480 (1975)
V. Vitek: Activities of muscle energy supplying enzymes after 5
days complete fasting in young men. Physiol. Bohemoslov.
(1982)31:311-314(1982) Requests for reprints should be addressed
to: MUDr. K. Vondra, CSc, Department of Medicine I of the Institute
for Clinical and Experimental Medicine, Videnska 800, 146 22 Prague
4 (Czechoslovakia) Horm. metabol. Res. 15 (1983) 271-274 Georg
Thieme Verlag Stuttgart New York Different Potencies of
Biosynthetic Human and Purified Porcine Insulin K.J. Schliiter, F.
Enzmann* and L. Kerp Zentrum fiir Innere Medizin, Abteilung fiir
Klinische Endokrinologie, Universitat Freiburg, and * E l i Lilly,
Bad Homburg, Germany Summary Introduction The glucose clamp
technique was used to compare the biological The biological
activity of insulin has been assessed by intra- activity of
purified porcine insulin and Biosynthetic Human venous bolus
injection, continuous infusion, and applica- Insulin (BHI). An
intravenous bolus of 0.1 U/kg BW was tion of a single subcutaneous
dose. Studies show that injected in eight male volunteers, and the
glucose was intravenous bolus injection produce counterregulatory
clamped at baseline values (euglycemic clamp). hormonal responses
which modify the metabolism of Serum insulin, serum C-peptide and
plasma glucose did not glucose. Since differences in the hormonal
responses to differ between porcine and human insulin. The insulin
in- duced glucose consumption differed significantly ( p <
0.007) human and porcine insulin have been reported, namely that
between purified porcine insulin (50.5 5.2 [SEM] g/2h) and human
insulin produced a lower output of growth hormone, Biosynthetic
Human Insulin (63.5 4 . 5 g / 2 h ) . Purified Cortisol, and
epinephrine (SchViter, Petersen, Borsche, porcine Insulin induced a
hormonal response w i t h significantly Hobitz and Kerp 1981;
SchViter, Petersen and Kerp 1982), ( p < 0.05) elevated
concentrations of serum growth hormone (12.1 0.25 ng/ml) and serum
Cortisol (161.4 28.6 ng/ml), it is difficult to compare the effects
on glucose metabolism. which were not observed following
Biosynthetic Human With the glucose clamp technique, essentially
normal fasting Insulin (serum growth hormone: 2.6 0.2 n g / m l ;
serum plasma glucose concentrations can be maintained (Pfeiffer,
Cortisol: 117.3 14.8 ng/ml). The data confirm earlier results
indicating hormonal and metabolic differences between Thum and
Clemens 1974; DeFronzo, Tobin and Andres human and porcine insulin.
1979;Nosadini, Noy, Kurtz and Alberti 1981). Hormonal responses are
minimal under these experimental conditions. Key-Words:
Biosynthetic Human Insulin Glucose Clamp The effect of Biosynthetic
Human Insulin (BHI) and puri- Technique Glucose Requirement Growth
Hormone fied porcine insulin (PPI) on glucose metabolism were meas-
Cortisol ured using the euglycemic clamp technique to avoid inter-
ference of other hormones. Subjects and Methods Informed consent
was obtained from 8 male volunteers (age: 24.2 1.3 years [mean
SEM], body weight: 75.1 1.3 kg; height: 185.6 Received: 15 March
1982 Accepted: 25 Aug. 1982 3.9 cm) without family or personal
history of diabetes. Laboratory
2. 272 Horm. metabol. Res. 15 (1983) K.J. Schliiter, F. Enzmann
and L. Kerp chemistry, ECG, physical examination and oral glucose
tolerance tests (100 g) were normal. In a randomized study, each
subject received both Biosynthetic Human Insulin (BHI) (Eli Lilly,
Indianapolis, CT 4969-1B) and identical formulated purified pork
insulin (PPI) (Eli Lilly, Indiana- polis, CT 4970-OA) intravenously
(bolus: 0.1 U/kg BW). The com- pound of BHI used in this experiment
was identical to the BHI (Lot 615-70N-174-9) used in the USP-rabbit
hypoglycemia assay. The biological activity in rabbits was 27.5 1.7
units/mg, which was 160 nmoles/mg for BHI and 159 nmoles/mg for
PPI. The tests were performed at one week intervals in this study.
One hour before the administration of insulin, three catheters were
in- serted into antecubital veins. The glucose-controlled insulin
infusion system (Biostator, Life Science Instruments, Miinchen,
FRG) was calibrated for continuous blood glucose monitoring and
glucose in- fusion. The plasma glucose concentration was clamped at
individual fasting baseline levels ( 0.25 mMol/1). The Biostator
was pro- grammed to maintain the individual (7696 mg/dl) euglycemia
of the subjects. Glucose (40 g/100 ml) was infused through the
three channels (saline-, glucose-, and optional channel) of the
Biostator. The maximum glucose infusion by the Biostator was 2.4
g/min. After an hour's rest, a bolus of insulin (0.1 U/kg BW) was
injected intravenously. Venous blood samples were obtained at-15,
0, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 75, 90, 105 and 120
minutes. Plasma glucose was determined by the glucose oxidase
technique with a Beckmann glucose analyzer (Beckmann Instruments,
Inc., Fullerton, Calif.). The following radioimmunoassays were
carried out: serum insulin (Phadebas-Insulintest,
Pharmacia-Diagnostics, AB, Uppsala, Sweden) (the cross-reaction of
PPI and BHI with the antibody used in this RIA was identical in the
range from 10 MU/ml to 320 MU/ml), serum Fig. 1 Plasma glucose
(mg/ml), serum insulin (MU/ml), serum C-peptide (Riamat, C-peptide
assay, Byk-Mallinckrodt, Diezenbach, C-peptide (ng/ml) after an
i.v. bolus (0.1 U/kg BW, given at 0 FRG), serum Cortisol
(Cortisol-Ria, Travenol, Cambridge, Mass.), minutes) of
Biosynthetic Human Insulin ( ) and purified and serum growth
hormone (Serono, Freiburg, FRG). pork insulin (oo) during
euglycemic clamp in eight male Interassay variations were reduced
by using the same immunoassay volunteers. The values at each time
interval represent the mean for all samples of an individual
subject. Intra-assay error, measured SEM of eight samples. There
was no statistical significance of as coefficient of variation, was
below 5% in all cases. differences between the porcine and human
group. The data are expressed as mean SEM. Areas under the
concentra- tion-time curves were calculated according to the
equation: *f ( x n+l- x n> (vn+l + yn) L Wilcoxons test for
paired differences was used. Results Eight male volunteers received
either Biosynthetic Human Insulin (BHI) (0.1 U/kg BW) or purified
porcine insulin (PPI) during an euglycemic clamp study. Plasma
glucose, serum insulin, serum C-peptide, serum growth hormone, and
serum Cortisol concentrations obtained from the venous blood
samples are shown in Fig. 1 and Fig. 2. The individual fasting
plasma glucose values (BHI: 89.1 1.6 mg/dl;PPI: 90.1 2.7 mg/dl)
were maintained through- out the clamping procedure, but those
following Bio- synthetic Human Insulin were slightly lower than
those following purified porcine insulin. Throughout the experi-
ment the maximum blood glucose fluctuation was 20 mg/dl and the
coefficient of variation of the plasma glucose values Fig. 2 Serum
growth hormone and serum Cortisol after an i.v. was below 15%.
bolus (0.1 U/kg BW, given at 0 minutes) of Biosynthetic Human
Insulin ( ) and purified porcine insulin (oo) during Serum insulin
concentrations did not differ during the test euglycemic clamp in
eight male volunteers. The values at each time period. Following
the intravenous administration identical interval represent the
mean SEM of eight samples. X, values that differ significantly from
the human insulin group at peak values (BHI: 309.3 61.0 juU/ml;
PPI: 335.2 64.2 that time interval (p< 0.05 by paired Wilcoxon's
test). /xU/ml) of insulin were obtained.
3. Potencies of Biosynthetic Human and Purified Porcine Insulin
Horm. metabol. Res. 15 (1983) 273 Serum C-peptide levels following
Biosynthetic Human Chiasson, Keller and Rubenstein 1978; Beischer,
Schmid, Insulin and purified porcine insulin did not differ
signifi- Kerner, Keller and Pfeiffer 1978) may be due to this tech-
cantly. A suppression of endogenous insulin secretion by nical
difference. exogenous insulin was not observed (Fig. 1). A
significant A small but significant rise in serum growth hormone
and (p< 0.05) elevation of serum growth hormone concentra- serum
Cortisol concentrations was observed following the tion (from 2.8
1.7 ng/ml at 0 minutes to 12.1 6.2 ng/ml injection of a bolus of
purified porcine insulin, which was at 30 minutes) occurred after
the administration of puri- not observed after Biosynthetic Human
Insulin. This can- fied porcine insulin. BHI did not produce any
fluctuation not be attributed to differences in plasma glucose or
to of serum growth hormone (2.5 1.9 ng/ml at 0 minutes plasma
glucose concentrations because plasma glucose was and 2.5 2.0 ng/ml
at 30 minutes). clamped at individual fasting glucose levels. The
secretion Serum Cortisol levels were also elevated (A serum
Cortisol of growth hormone and of Cortisol appears to be a response
38.8 8.1 ng/ml) after purified porcine insulin but not to the
heterologous insulin. after human insulin (A serum Cortisol 23.1
4.5 ng/ml) Significantly more exogenous glucose was required to (p.
Low bicarbonate levels 1979). However, only the influence of age
and diabetes increased the absorption ( T 1 / 2 3.92.3 h , p <
0.05). Lean duration on NPH insulin absorption have been
systematical- diabetics had a faster absorption (6.2 1.9 h) than
normal ly studied (Dobson, Robbins, Johnson, Mdalel, Odem, Corn-
weight diabetics (7.5 2.0 h, p < 0.02). Sex, age, diabetes wall
and Davis 1967). The aim of this study was to assess duration and
injection depth d i d not influence T 1 / 2 . The some clinical
factors which might influence the absorption half life was
significantly inversely correlated to the resting subcutaneous
blood f l o w (r= - 0 . 8 8 2 , p < 0.01). The overall of
I25I-NPH insulin from the subcutaneous tissue in diabetic
interindividual coefficient of variation for insulin absorption
patients by using the biotelemetric technique. The absorp- in
nonketotic diabetics was 27.4%. Also considerable intra- tion of
insulin was measured to determine the influence patient day-to-day
variation was found (24.5%), and between of lipohypertrophy,
ketosis, body weight, sex, age, dia- different injection sites
(30.2%). These variations emphasize betes duration and inter- and
intrapatient variation. In some the drawbacks of conventional
insulin therapy in the manage- ment of insulin-requiring diabetics.
diabetics, also the effect of injection depth and subcutane- ous
bloodflow(SBF) was evaluated. Key-Words: Insulin Absorption NPH
Insulin Diabetes Mellitus Lipohypertrophy Ketosis Body Weight Sex
Age Diabetes: Duration Subcutaneous Blood-Flow Material and Methods
125 l-NPH Insulin One hundred and one diabetic patients were
investigated as inpatients in 194 studies after informed consent
was obtained. Group 1 com- prised 20 patients with
insulin-dependent diabetes mellitus (IDDM) Received: IS March 1982
Accepted: 31 July 1982 with palpable lipohypertrophy at the
injection site. Group 2 included