Arild E. Hansen

Study of Certain Tissue Lipids in Generalized Lipodystrophy (“Lipohistiodiaresis”) .∗

An unusual opportunity to study the lipid composition of various tissues in an extremely rare condition, that of generalized lipodystrophy, was offered when death occurred in a 9-year-old boy in whom an almost complete absence of adipose tissue from the body had been present for the past 6 years (Case 1). In addition to this remarkable apparent lack of body fat, the symptom complex was composed of cirrhosis of the liver, chronic fibrosis of the spleen, pancreas and certain lymph nodes, and diabetes mellitus. Necropsy was begun within one hour following demise, at which time samples of various tissues were obtained. After being weighed, the specimens (usually about 1 g of tissue) were placed in 95% alcohol and allowed to stand for 24 hours. The tissues were then ground with sea sand in a mortar, rinsed several times with alcohol and ether, returned to the original flasks, and sufficient ether added to make approximately a 3:1 alcohol-ether mixture. The flasks were immersed in a boiling water bath for about 5 minutes and allowed to cool; the contents were filtered through fat-free filter paper into volumetric flasks, brought to volume, and stored in a refrigerator until analyses were made. Aliquots were measured and the following procedures employed: The method of Wilson and Hansen 1 was used for the determination of the unsaponifiable and saponi-fiable fractions, while the technic followed by Hansen 2 was used for the determination of the fatty acids in the acetone-insoluble (phospholipid) fraction and the acetone-soluble (cholesterol ester-neutral fat) fraction. The total cholesterol and cholesterol esters were determined by the procedure described by Bloor, 3 4 the photoelectric colorimeter being used in obtaining the final readings. For the control studies, similar tissues from a 14-vear-old boy dying in uremia from subacute nephritis (Case 2), the best available material at the time, were used.

Serum Lipid Changes and Therapeutic Effects of Various Oils in Infantile Eczema

In a previous communication 1 the iodine number of the serum fatty acids was reported to be abnormally low in infantile eczema during the active phase of the disease. Additional data have confirmed the original findings. The average iodine number of the serum fatty acid in 21 determinations on 11 cases of infantile eczema was 82, while that of 21 determinations on 11 normal infants in the same age group was 114. The magnitude of the difference between these 2 groups is similar to that between rats suffering from unsaturated fatty acids deficiency and normal rats. 2 In December, 1931, oils especially rich in unsaturated fatty acids were given to 2 infants suffering with severe eczema, with good clinical results. Twelve additional cases treated in like manner since that time have shown a similar response. This paper reports changes in the iodine number of the serum fatty acids in 4 cases which could be kept under close observation over a period of months while receiving treatment. All 4 received the same diet and same local care of the skin. Three received supplements of linseed or corn oil while the fourth received none. The iodine number of the serum fatty acids was subnormal in all 4 cases before treatment was instituted, but increased to normal levels, in cases RS, DL and BS coincidentally with the marked clinical improvement. (Fig. 1.) In contrast to this response was that of the infant RB. This control subject, receiving no oil supplement, failed to show definite clinical improvement. The iodine number not only did not rise during the long period of observation but actually fell slightly. Infant DL was carefully observed in the hospital for about 3 months for any sign of a spontaneous remission before being given oil. No improvement occurred during this period and the iodine number remained low. Three weeks after oil was added to the diet, however, the skin became clear and the iodine number rose to the normal level as it had done in the other 2 treated cases.

Study of Iodine Number of Serum Fatty Acids in Infantile Eczema

The observation that marked skin changes develop in rats suffering from the unsaturated fatty acid deficiency disease described by Burr and Burr 1 suggested the possibility that certain dermatological disorders of childhood, such as infantile eczema, might be dependent in part at least upon this type of dietary deficiency. A few preliminary observations on eczematous babies given liberal supplements of unsaturated fatty acids in their diets in addition to the routine treatment were sufficiently encouraging to suggest the desirability of careful study of the blood lipids in this disease. In order to determine whether any relationship might exist between the two clinical entities, similar blood studies were undertaken on eczematous infants and on rats suffering from the unsaturated fatty acid deficiency. The present paper deals exclusively with the results in the cases of infantile eczema. The total iodine absorption of the serum was first determined on one or more occasions in 10 cases of eczema and in 16 normal infants of similar age. This was followed by determinations of the total fatty acids, cholesterol, total iodine absorption of the serum and the iodine number of the serum fatty acids in 5 normal and 6 eczematous infants. All blood samples were collected between 12 and 16 hours after a meal. Bloors methods 2 were used to determine the cholesterol and total fatty acids in the serum. The Rosenmund-Kuhnhenn 3 method as modified by Page, Pasternak and Burt4 was used to determine the iodine absorption of the serum lipids. The preliminary data on the total iodine absorption values of the sera are as follows: The average for 21 determinations on 10 infants with eczema was 383 mg. (range 280-487) iodine absorbed per 100 cc. serum.

Arachidonic and Linolic Acid of the Serum in Normal and Eczematous Human Subjects.

Summary and Conclusions Linolic acid (tetrabromides calculated as sudh) is present in human serum to the extent of about 5% of the total fatty acids, and arachidonic acid (polybromides calculated as such) about 37% of the total fatty acids in pooled samples of blood serum from normal children. The content oi both these fatty acids is definitely diminished in children with eczema. In those cases of active eczema which were clinically cured either by the internal administration of oils rich in the unsaturated fatty acids or by the local application of ointments containing crude coal tar, or both, the arachidonic acid was not increased while the linolic content was the same 2s that found in normal subjects. These findings indicate a possible reason for the low iodine number of the serum fatty acids in eczema and further suggest that a disturbance in the metabolism of the unsaturated fatty acids may be one of the many factors at fault in this condition.

Studies With Dogs Maintained on Diets Low in Fat.

Conclusions Distinct alterations in the appearance of the skin have been observed in puppies reared on a diet low in fat. This skin change became evident at about 3 months of age with the gradual development of a generalized flaky desquamation together with a dryness and coarseness of the hair. The skin and hair of the litter-mate puppies receiving 28% of their calories as lard in the diet remained clear and soft. Coincidental with this phenomenon marked differences in the degree of unsaturation of the fatty acids of the blood serum in the 2 groups of animals was demonstrated. The most marked difference in the iodine number of the fatty acids was found in the acetone soluble fraction, the average value being 83.9 for the animals receiving practically no fat in contrast to 1 18.7 for the animals receiving lard as a form of fat in the diet.

Iodine Numbers of Serum Lipids in Rats Fed on Fat-Free Diets.

Summary (1) The cholesterol and total fatty acids of the serum in rats fed on a fat-free diet were found to be lower than in control animals. (2) The iodine number of the serum fatty acids from rats fed on fat-free diets indicates that the fatty acids are less unsaturated than those of controls.

Study of the serum lipids in the celiac syndrome

Summary 1. Ten children with the celiac syndrome were seen in the Department of Pediatrics at the University of Minnesota Hospitals from September, 1940, to August, 1942. Six of these patients died, in four of whom cystic fibrosis of the pancreas was demonstrated at autopsy. In the fifth and sixth patients autopsy was not performed, but the clinical symptoms and the laboratory findings suggested the same condition for at least one of them. Of the four children still living, one is considered to have a cystic fibrosis of the pancreas because of the abnormal response to the injection of secretin as well as the clinical nature and laboratory findings and family history. The other three living children with symptoms typical of the celiac syndrome left the hospital considerably improved. These patients are still under observation and the cause of their disturbance is still obscure. 2. In the eight patients in whom the serum lipids were studied, the values for the various fractions of the serum lipids (total fatty acids, acetone-soluble fatty acids, acetone-insoluble fatty acids, total cholesterol, cholesterol esters, and free cholesterol) were within the normal range of variability, except in one patient who had a low value for total cholesterol. 3. The iodine numbers of the fatty acids of all fractions of the lipid in these eight patients were abnormally low. 4. In five instances no increase in the serum lipids occurred following the ingestion of a fatty meal; in one patient there was a slight increase; in two the increase was normal. It happened that the latter two were considered unlikely to have a cystic fibrosis of the pancreas. 5. On two occasions no better absorption of fat was observed when the fatty meal consisted of a fat (corn oil) of a high degree of unsaturation. 6. The low degree of unsaturation of the serum fatty acids in the patients with celiac syndrome seems to be best explained on the basis of a deficient fat absorption. From these studies it is suggested that the human organism is unable to synthesize the more highly unsaturated fatty acids. 7. It appears that in patients with the celiac syndrome, the determination of the iodine numbers of the serum fatty acids might offer a means of ascertaining their ability to absorb fats. Since the steatorrhea, ordinarily present in patients with this condition is often lacking when the patient has been maintained on the usual relatively low fat diet, this might offer a relatively simple but helpful procedure in confirming the diagnosis and in evaluating clinical improvement in these patients. 8. The deficient fat absorption in the celiac syndrome seems to be difficult to explain with the present knowledge of the physiology of fat metabolism.

Possible Mechanism of Crude Coal Tar Therapy in Infantile Eczema.

The therapeutic value of ointments containing crude coal tar in infantile eczema has long been recognized. It is not infrequently noted in eczema cases showing extensive skin eruption that untreated areas, as well as those to which direct application of the tar is made, improve. This suggests that crude coal tar may have a systemic as well as a local effect. In the light of the observations that the serum fatty acids of infants with eczema are less unsaturated than those of normal subjects of the same age 1 and that administration of oils high in unsaturated fatty acids causes an increase in the degree of unsaturation as well as a clinical improvement in the condition, 2 a preliminary study of the effect of local applications of tar ointment on the fatty acids of the serum has been made. Data from 3 cases followed over a period of several months are presented here. Case 1, an eczematous male infant 15 months of age, showed values of 334 and 322 mg. iodine absorbed per 100 cc. of serum on 2 successive days at the height of an outbreak of eczema. Eight days after local treatment with crude coal tar ointment the patient had improved clinically and the iodine absorption had risen to 486. One and 2 months later the values were 494 and 520 respectively and the condition remained good. In subsequent cases the iodine number of the serum fatty acids was studied. Case 2, an infant 5 months of age with severe eczema, was found during the active phase of the disease to have an iodine number of the serum fatty acids ranging between 79 and 81. After application of coal tar ointment for one week, which caused marked improvement in the condition of the skin, the iodine number was found to be 109. At the end of the second week of treatment it was 95 and two weeks later was 101. Active treatment was then omitted for 3 weeks, during which the skin remained entirely clear, except for the appearance of a slight eruption in the folds of the neck. The iodine number at the end of this period was 113. Case 3, a 6-months-old infant with severe eczema showed an iodine number of the serum fatty acids of 78 before the ointment was applied. This had risen to 96 at the end of a 10-day period of treatment during which the lesions showed marked improvement. Four weeks after the tar was stopped the iodine number had again fallen to 84 and in 5 weeks it was found to be 76.

Nature of Fatty Acids of Acetone Insoluble (Phospholipid) Fraction of Serum.

Summary This microgravimetric technic for the study of serum lipids in 14 normal infants and children disclosed that the acetone insoluble (phospholipid) fatty acids comprised about one-third of the total fatty acids. These fatty acids were more unsaturated and also had higher molecular weights than did the total fatty acids. Most methods used for the estimation of fatty acids of phospholipid fraction assume the presence of 18 carbon atoms. Our finding of high molecular weights indicates the necessity of revision of the calculations used in their determination.

Phosphatase Activity of the Serum and Tissues in Osteogenesis Imperfecta

Robison 1 has pointed out that the enzyme phosphatase is an important factor in the calcification of bone. Kay 2 has more recently summarized the evidence for its activity in connection with studies on a wide variety of clinical conditions. Since osteogenesis imperfecta is an outstanding example of defective bone formation for which no pathogenesis has as yet been discovered, a comprehensive study of the various metabolic factors which might be involved in this condition was begun by the writer in 1931. The present report deals with the phase of that study which concerns the phosphatase activity of the blood serum and fixed tissues. Kay reported the serum phosphatase activity to be slightly increased in some cases of osteogenesis imperfecta, while Bodansky and Jaffe 3 found it to be normal in 6 cases. Periodic determinations of the serum phosphatase by the method of Kay have now been made in 4 cases of osteogenesis imperfecta under a variety of experimental conditions. In one case the fixed tissue phosphatase has also been studied postmortem. Repeated determinations in the 4 cases showed values almost identical with those found for normal control subjects and for a miscellaneous group of patients in the same age period. Variations in the ordinary constituents of the diet were found to have no definite effect on the serum phosphatase values. Following the administration of vitamin D in relatively enormous doses in the form of viosterol 10,000X, however, the phosphatase activity of the serum was uniformly reduced. Parathormone administration over a period of several days had a similar but less marked depressing effect on the enzyme.