Hum. Hered. 28: 285-292 (1978)
ABO Blood Groups and Serum Proteins in Thromboangiitis obliterans (Buerger’s Disease) R .L . G u pta a n d S .S .P apiha Department of Surgery, Himachal Pradesh Medical College, Simla, and Department of Human Genetics, University of Newcastle upon Tyne, Newcastle upon Tyne
Key Words. Thromboangiitis obliterans • Buerger’s disease • Serum proteins ■ABO blood groups • Albumin/globulin ratio
Differential diagnosis of peripheral atherosclerotic obliterative disease and thromboangiitis obliterans (TAO) was for long the subject of debate. The latter condition lacked definite biochemical identity and so the condition has to be defined on clinical observations alone. W essler et al. [1960] indeed felt that the two were different forms of the same condition but in recent years associated haematological variations have shown that the two are distinct entities. C raven and C otten [1968] showed a higher concentration of heparin-precipitable fractions of fibrinogen in the plasma of patients with Buerger’s disease, and highly positive correlations of this both with fibrino gen and erythrocyte sedimentation rate. Such a correlation was not seen in atherosclerosis. Biochemical distinction was also observed in the levels of Nacetylneuraminic acid (NANA) [W alaszew sk a , 1969]; patients with peri pheral obliterative atherosclerosis showed higher levels of NANA in all age groups (20-70 years), whereas in TAO patients the level of NANA remained normal. Serum proteins are frequently used for routine analysis in various diseases, and one object of the present study was to examine the changes in serum proteins that occur in TAO disease.
Downloaded by: Nagoya University 133.6.82.173 - 1/13/2019 12:26:29 PM
Abstract. In Madhya Pradesh, India, 43 patients with thromboangiitis obliterans showed a decrease in total serum proteins and in the albumin fraction, an increase in a,and a2-globuIm fractions, and decrease in the albumin/globulin ratio. In the ABO blood groups, patients show a significant excess of group B individuals. It is possible that the disease may be more severe in individuals with the B blood group substance.
286
G upta /P apiha
The aetiology of TAO disease is complex, and as in some other arterial diseases, how far the individual’s constitution plays a role in its development is not clear. That genetic factors may be involved in atherosclerosis is suggested by the studies of J aegerm an [1962] and K in g sbu ry [1971] who showed a possible association with blood group A; however, A lla n [1973] was not able to confirm this association. There is little direct evidence for the role of heredity in TAO, though Z a n n in i and C o tru po [1973] reported this disease to follosv a similar course in two brothers. Our present investigation was initiated with two objectives, firstly to examine the quantitative changes of serum proteins in patients with different degrees of severity of the disease, and secondly to enquire whether there is any association with the ABO blood groups in Buerger’s disease such as has been suggested in atherosclerosis.
Methods and Material 43 patients under the age of 40 years front different regions of the Indian state of Madhya Pradesh were diagnosed and accepted as suffering from TAO by the clinical criteria of intermittent claudication, history of smoking, absence of arterial pulsation and manifestation or threat of gangrene. All patients included in this survey manifested the above four criteria. Routine and special examinations eliminated other causes of gangrene, as well as such conditions as diabetes and hyperlipidaemia which were also excluded by routine biochemical analysis. For comparison, a control group was obtained from normal healthy laboratory and hospital workers of the same ages from the same regions. Paper electrophoresis was performed in Barbitone buffer, pH 8.6, by using freely hanging vertical strips with a voltage not exceeding 100-150 V, subsequently stained with bromophenol blue, following the method of Smith [1968]. Quantitation of the scrum proteins was performed by scanning the filter strips in an extinction recording instrument Carl Zeiss model E R 1-10. Total proteins were estimated by the method of Micro-Kjeldahl. ABO blood grouping was performed by standard tile techniques.
The patients were divided into 3 groups according to the extent of limb involvement: group 1: patients with one lower limb involved; group 2: patients with both lower limbs involved; group 3: patients with all four limbs involved. The results for the serum protein fractions are shown in table I.
Downloaded by: Nagoya University 133.6.82.173 - 1/13/2019 12:26:29 PM
Results
Number of Total protein Albumin“ individuals tested
No. 1 22 No. 2 13 No. 3 8 F (d.f. 2, 40) Total patients, N o .( l+ 2 + 3)43 Normal 20 t (d.f. 61)
a,-Globulina a2-Globulina /9-Globulin3
/-Globulin3
Albumin/globulin ratio
6.09±0.71 5.07+0.90 5.27+0.43 9.04”
42.91+3.34 35.70+5.62 41.35+3.45 9.62”
8.91+2.01 8.20+1.74 8.25+1.28 0.78*
15.86+3.63 15.40+1.76 15.85+3.41 0.10b
11.91+2.49 12.70+2.13 14.10+3.83 1.99b
21.41+4.77 28.00+7.63 20.40+6.30 5.96*
0.72+0.11 0.59+0.13 0.70+0.18 6.11*
5.48+0.68 7.22+0.71 9.32”
39.65+4.14 51.30+2.97 11.29”
8.45+1.68 4.79+1.23 8.70”
15.70+2.93 9.00+1.61 9.55”
12.90+2.81 11.78+1.84 1.62b
23.27+6.20 23.13+2.96 0.10b
0.67+0.11 1.08+0.13 13.06”
a Values expressed as percent of total serum protein. b Not significant. * Significant at 0.01; *’ significant at 0.001.
Downloaded by: Nagoya University 133.6.82.173 - 1/13/2019 12:26:29 PM
Group of patients
Buerger’s Disease, Blood Groups and Serum Proteins
Table l. Serum protein fractions in three groups of patients and controls (means +SD )
G upta/P apiha
Since the distribution of the quantitative values among the various groups appeared normal, intercomparison of the various groups could be made directly by the t or the F test. The average values for the normal control group of 20 individuals showed no significant differences from previously reported values for the Indian region [G h ai and K u m a r , 1965], But the total group of the patients with TAO showed several changes in serum proteins: i) A significant decrease of some 26% was seen in total serum proteins compared to the normal value. ii) The albumin fraction of 40% was also significantly decreased com pared to the normal value of 51%. iii) In the globulin fractions there was a significant increase in ax and a2 fractions, respectively. There was no increase in the y-globulin fraction and no significant increase in the /1-globulin fraction; neither can be regarded as showing any deviation from the normal values. iv) There was a significant decrease of the albumin/globulin ratio in these 2 groups. By comparison with the normal group, all 3 subgroups of patients showed a significant decrease in total serum proteins and in albumin, and a marked increase in the eq- and a2-globulin fractions. Intercomparison of the 3 groups showed some variation from one to another in the different fractions highly significant in total protein, albumin and y-globulin and the albumin/globulin ratio, but none showed any clear trend with increasing severity. Table II shows the ABO blood group results of the 43 patients with TAO. The number with blood group B is remarkable. The observed numbers do not show any significant deviation from those expected from the gene fre quencies, so that technique and sampling are not the explanation. As a normal sample, data from two different studies of the Madhya Pradesh population were pooled [R oberts et ah, 1974; K um ar and G h o sh , 1967], While this too shows a high B blood group frequency it is by no means as elevated as in the patients. Comparison with this showed a significant increase of individuals with the B gene and decrease of blood group O indivi duals in the TAO patients (A'2 = 13.40, p
ABO Blood Groups and Serum Proteins in Thromboangiitis obliterans (Buerger’s Disease) R .L . G u pta a n d S .S .P apiha Department of Surgery, Himachal Pradesh Medical College, Simla, and Department of Human Genetics, University of Newcastle upon Tyne, Newcastle upon Tyne
Key Words. Thromboangiitis obliterans • Buerger’s disease • Serum proteins ■ABO blood groups • Albumin/globulin ratio
Differential diagnosis of peripheral atherosclerotic obliterative disease and thromboangiitis obliterans (TAO) was for long the subject of debate. The latter condition lacked definite biochemical identity and so the condition has to be defined on clinical observations alone. W essler et al. [1960] indeed felt that the two were different forms of the same condition but in recent years associated haematological variations have shown that the two are distinct entities. C raven and C otten [1968] showed a higher concentration of heparin-precipitable fractions of fibrinogen in the plasma of patients with Buerger’s disease, and highly positive correlations of this both with fibrino gen and erythrocyte sedimentation rate. Such a correlation was not seen in atherosclerosis. Biochemical distinction was also observed in the levels of Nacetylneuraminic acid (NANA) [W alaszew sk a , 1969]; patients with peri pheral obliterative atherosclerosis showed higher levels of NANA in all age groups (20-70 years), whereas in TAO patients the level of NANA remained normal. Serum proteins are frequently used for routine analysis in various diseases, and one object of the present study was to examine the changes in serum proteins that occur in TAO disease.
Downloaded by: Nagoya University 133.6.82.173 - 1/13/2019 12:26:29 PM
Abstract. In Madhya Pradesh, India, 43 patients with thromboangiitis obliterans showed a decrease in total serum proteins and in the albumin fraction, an increase in a,and a2-globuIm fractions, and decrease in the albumin/globulin ratio. In the ABO blood groups, patients show a significant excess of group B individuals. It is possible that the disease may be more severe in individuals with the B blood group substance.
286
G upta /P apiha
The aetiology of TAO disease is complex, and as in some other arterial diseases, how far the individual’s constitution plays a role in its development is not clear. That genetic factors may be involved in atherosclerosis is suggested by the studies of J aegerm an [1962] and K in g sbu ry [1971] who showed a possible association with blood group A; however, A lla n [1973] was not able to confirm this association. There is little direct evidence for the role of heredity in TAO, though Z a n n in i and C o tru po [1973] reported this disease to follosv a similar course in two brothers. Our present investigation was initiated with two objectives, firstly to examine the quantitative changes of serum proteins in patients with different degrees of severity of the disease, and secondly to enquire whether there is any association with the ABO blood groups in Buerger’s disease such as has been suggested in atherosclerosis.
Methods and Material 43 patients under the age of 40 years front different regions of the Indian state of Madhya Pradesh were diagnosed and accepted as suffering from TAO by the clinical criteria of intermittent claudication, history of smoking, absence of arterial pulsation and manifestation or threat of gangrene. All patients included in this survey manifested the above four criteria. Routine and special examinations eliminated other causes of gangrene, as well as such conditions as diabetes and hyperlipidaemia which were also excluded by routine biochemical analysis. For comparison, a control group was obtained from normal healthy laboratory and hospital workers of the same ages from the same regions. Paper electrophoresis was performed in Barbitone buffer, pH 8.6, by using freely hanging vertical strips with a voltage not exceeding 100-150 V, subsequently stained with bromophenol blue, following the method of Smith [1968]. Quantitation of the scrum proteins was performed by scanning the filter strips in an extinction recording instrument Carl Zeiss model E R 1-10. Total proteins were estimated by the method of Micro-Kjeldahl. ABO blood grouping was performed by standard tile techniques.
The patients were divided into 3 groups according to the extent of limb involvement: group 1: patients with one lower limb involved; group 2: patients with both lower limbs involved; group 3: patients with all four limbs involved. The results for the serum protein fractions are shown in table I.
Downloaded by: Nagoya University 133.6.82.173 - 1/13/2019 12:26:29 PM
Results
Number of Total protein Albumin“ individuals tested
No. 1 22 No. 2 13 No. 3 8 F (d.f. 2, 40) Total patients, N o .( l+ 2 + 3)43 Normal 20 t (d.f. 61)
a,-Globulina a2-Globulina /9-Globulin3
/-Globulin3
Albumin/globulin ratio
6.09±0.71 5.07+0.90 5.27+0.43 9.04”
42.91+3.34 35.70+5.62 41.35+3.45 9.62”
8.91+2.01 8.20+1.74 8.25+1.28 0.78*
15.86+3.63 15.40+1.76 15.85+3.41 0.10b
11.91+2.49 12.70+2.13 14.10+3.83 1.99b
21.41+4.77 28.00+7.63 20.40+6.30 5.96*
0.72+0.11 0.59+0.13 0.70+0.18 6.11*
5.48+0.68 7.22+0.71 9.32”
39.65+4.14 51.30+2.97 11.29”
8.45+1.68 4.79+1.23 8.70”
15.70+2.93 9.00+1.61 9.55”
12.90+2.81 11.78+1.84 1.62b
23.27+6.20 23.13+2.96 0.10b
0.67+0.11 1.08+0.13 13.06”
a Values expressed as percent of total serum protein. b Not significant. * Significant at 0.01; *’ significant at 0.001.
Downloaded by: Nagoya University 133.6.82.173 - 1/13/2019 12:26:29 PM
Group of patients
Buerger’s Disease, Blood Groups and Serum Proteins
Table l. Serum protein fractions in three groups of patients and controls (means +SD )
G upta/P apiha
Since the distribution of the quantitative values among the various groups appeared normal, intercomparison of the various groups could be made directly by the t or the F test. The average values for the normal control group of 20 individuals showed no significant differences from previously reported values for the Indian region [G h ai and K u m a r , 1965], But the total group of the patients with TAO showed several changes in serum proteins: i) A significant decrease of some 26% was seen in total serum proteins compared to the normal value. ii) The albumin fraction of 40% was also significantly decreased com pared to the normal value of 51%. iii) In the globulin fractions there was a significant increase in ax and a2 fractions, respectively. There was no increase in the y-globulin fraction and no significant increase in the /1-globulin fraction; neither can be regarded as showing any deviation from the normal values. iv) There was a significant decrease of the albumin/globulin ratio in these 2 groups. By comparison with the normal group, all 3 subgroups of patients showed a significant decrease in total serum proteins and in albumin, and a marked increase in the eq- and a2-globulin fractions. Intercomparison of the 3 groups showed some variation from one to another in the different fractions highly significant in total protein, albumin and y-globulin and the albumin/globulin ratio, but none showed any clear trend with increasing severity. Table II shows the ABO blood group results of the 43 patients with TAO. The number with blood group B is remarkable. The observed numbers do not show any significant deviation from those expected from the gene fre quencies, so that technique and sampling are not the explanation. As a normal sample, data from two different studies of the Madhya Pradesh population were pooled [R oberts et ah, 1974; K um ar and G h o sh , 1967], While this too shows a high B blood group frequency it is by no means as elevated as in the patients. Comparison with this showed a significant increase of individuals with the B gene and decrease of blood group O indivi duals in the TAO patients (A'2 = 13.40, p
