Hans Kollberg

The Reserpinized Rat in the Study of Cystic Fibrosis: X-ray Microanalysis of Submandibular Gland and Pancreas

The chronically reserpinized rat has been suggested as an animal model for cystic fibrosis. X-ray microanalysis of thick and thin cryosections was carried out to assess elemental redistribution in the submandibular glands and the pancreas of reserpinized rats at the cellular and subcellular level. In the submandibular gland of reserpinized rats, calcium and magnesium concentrations were significantly elevated. Mucus globules, secretory granules, and endoplasmic reticulum were the primary sites of the localization of excess calcium and magnesium. A significant potassium loss from the gland had occurred, particularly from the serous cells. Electron microscopy of conventionally prepared tissue showed marked swelling of the endoplasmic reticulum, especially in mucous cells. The elemental changes in the pancreatic acinar cells of reserpinized rats were reminiscent of elemental redistribution connected with cell death: increased levels of sodium, chlorine, and calcium and decreased levels of magnesium and potassium. Ultrastructural changes included swelling of the endoplasmic reticulum and obstruction of the acinar lumen. It is concluded tha elemental redistribution in chronically reserpinized rats presents interesting parallels with cystic fibrosis.

INCIDENCE AND SURVIVAL CURVES OF CYSTIC FIBROSIS IN SWEDEN

ABSTRACT. A registry of all patients with cystic fibrosis (CF) in Sweden has been kept since 1968. During the 10‐year period 1968‐1977 the number of CF patients in Sweden increased by about 8‐9 per year, i. e. from 116 to 200, the median age of patients increased from 5 to 11 years, the percentage of patients above 18 years of age increased from 3 to 9 of the total CF population, and the median survival age increased from 7.5 to 16 years. For CF patients born without meconium ileus the median survival age in 1973‐1977 was 17.8 years. From these figures the incidence of CF in Sweden is estimated to be 1:2200 to 1:4500. Several factors may have contributed to the improved outcome for CF patients, such as better medical treatment with more intensive physical therapy, the use of inhalation therapy and new antibiotics, improved social care and intensified information.

Electrolyte redistribution in cystic fibrosis fibroblasts studied by electron probe X-ray microanalysis.

The elemental composition of fibroblasts from cystic fibrosis (CF) patients and from healthy controls was compared by means of electron probe X-ray micro-analysis. Significantly lower sodium levels (p < .01) and higher calcium levels (p < .02) were found in CF fibroblasts, indicating a disturbance of ion regulation in these cells. Treatment with Staphylococcus aureus protein A and gammaglobulin (proteins that may influence the ciliotoxic CF factor) did not change the elemental composition of the fibroblasts.

Acid hydrolases in sera and plasma from patients with cystic fibrosis.

The enzyme activities of alpha-fucosidase (pH 4.0 and pH 5.5), alpha-galactosidase, beta-galactosidase, alpha-glucosidase (pH 4.5 and pH 6.0), beta-glucosidase, beta-glucuronidase, beta-hexosaminidase, and alpha-mannosidase (pH 4.5 and pH 5.5) were investigated in sera from cystic fibrosis (CF) patients. Several of these activities were significantly increased in sera from patients compared to age-matched control children. CF-patients in a more advanced stage of the disease had a tendency to higher values of some of these hydrolases than those in better condition. No new isoenzymes of these hydrolases were found. Only minor differences could be detected in the pH-profiles of alpha-mannosidase and acid phosphatase from age-matched normal controls, heterozygotes and homozygotes for CF. With our technique, alpha-mannosidase and acid phosphatase showed the same thermostability in CF-patients. CF-heterozygotes and age-matched controls, except at 56 degrees C, when the activity of acid-phosphatase in the plasma from adult CF-heterozygotes decreased more than that from adult controls

Ribonuclease in Different Types of Saliva from Cystic Fibrosis Patients

ABSTRACT. The flow rate, the activity of ribonuclease (RNAase) and the concentration of protein were determined in whole saliva and in parotid and submandibular saliva from patients with cystic fibrosis (CF) and from healthy controls, both before and after stimulation of the salivary secretion. Lower flow rates were found in all types of saliva from CF‐patients than in control saliva. Increased activity of RNAase was found in CF saliva, both before and after stimulation of the secretion. The concentration of protein was also increased, but to a lesser degree. No correlation was found between either RNAase activity or protein concentration and severity of the disease or age of the children. It is therefore unlikely that these disturbances are secondary to progression of the disease. Considerable variations in RNAase activity and protein concentration were observed, especially within the CF group, and therefore despite the significant increase in these variables in CF their estimation is of limited diagnostic value.

RELIABLE DIAGNOSIS OF THE MAJOR TYPE OF CYSTIC FIBROSIS WITH FIBROBLAST CULTURES A Double Blind Study

ABSTRACT. A double blind study has been carried out to demonstrate that the most common type of cystic fibrosis (CF) can be reliably diagnosed with skin‐derived fibroblast cultures. Alkaline phosphatase (AIP) activity has been measured in 6 normal controls, 12 CF‐heterozygotes and 6 CF‐homozygotes before and after stimulation with Tamm‐Horsfall‐glycoprotein (THP), isoproterenol and theophylline (“THP‐induction test”). The mean AIP‐activities after THP‐induction were 8.8, 12.7 and 34.6 for the three different genotypes respectively. There was no overlap between the values of CF‐homozygotes on the one hand, and the values of CF‐heterozygotes and normal controls on the other hand. All 24 specimens were correctly diagnosed in the present double blind study, indicating the very high degree of reliability of the THP‐induction test in the detection of the predominant type of cystic fibrosis with fibroblast cultures. Normal controls and CF‐heterozygotes could not be discriminated on an individual basis, but as a group the CF‐heterozygotes displayed higher AIP‐values.

Effect of chronic treatment with cystic fibrosis fibroblast medium on rat submandibular gland acinar cells.

The effect of chronic treatment with cystic fibrosis (CF) fibroblast medium on rat submandibular gland and pancreas was investigated. Rats were injected for 8 days with conditioned medium from normal or CF fibroblasts. The elemental content of the acinar cells was measured by X-ray microanalysis of cryosections. A significant increase in cellular calcium, and a decrease in cellular sodium concentrations were found after treatment with CF medium. The ultrastructure of the submandibular acinar cells was not affected by the conditioned CF fibroblast culture medium. No effect of treatment with CF medium on ultrastructure and elemental content of pancreatic acinar cells could be demonstrated. The response to alpha-adrenergic, beta-adrenergic, cholinergic, and peptidergic stimulation in submandibular gland acinar cells of rats injected with normal or CF medium was investigated in vitro. With regard to changes in elemental composition after stimulation, no significant differences in response between the two groups could be found. Apparently, a factor in conditioned medium from cultured CF fibroblasts induces a net increase in calcium content of rat submandibular gland acinar cells. Possibly, this factor acts in a similar way in CF patients and may cause elevated calcium levels in CF cells.

Diagnosis of cystic fibrosis homozygotes and heterozygotes from plasma and fibroblast cultures. A three-generation family study

The diagnosis of cystic fibrosis (CF) homozygotes and heterozygotes and of individuals without the CF gene, based on differences in the thermal stability of acid phosphatase and a‐mannosidase, is reported. The residual activities at 36.5°C and 41.3°C were below 10% of the activity in unheated samples for homozygotes, 4&50% for heterozygotes and above 90% for normals. The intracellular alkaline phosphatase and extracellular β‐hexosaminidase activities after treatment with heparin and gammaglobulin were 500% and 200%, respectively, of the activities without this treatment in CF homozygotes, whereas for heterozygotes and normals the values were the same after treatment as before. Pedigrees of four CF families, covering 2–3 generations, are presented. The possible use of these tests as diagnostic tools is further discussed.

CYSTIC FIBROSIS-LIKE CHANGES IN BODY FLUIDS OF HEALTHY PERSONS PERFORMING ANAEROBIC EXERCISE

Ceder, O., Teien, D., Bardoń, A., Hellsing, K. and Kollberg, H. (Department of Pediatrics, University Hospital, Umeå, Sweden, Department of Clinical Physiology, University Hospital, Umeå, Sweden, Department of Clinical Chemistry, University Hospital, Uppsala, Sweden, Department of Biochemistry, Institute of Sport, Warsaw, Poland and Department of Pediatrics, University of Kuwait, Safat, Kuwait). Cystic fibrosis‐like changes in body fluids of healthy persons performing anaerobic exercise. Acta Paediatr Scand 1983, Suppl. 309: 25. The biochemical compositions of blood, saliva and urine were compared at rest and during and after anaerobic physical exercise in homozygotes and heterozygotes for cystic fibrosis (CF) and in healthy controls.

EFFECTS OF SUCROSE LOAD ON THE BIOCHEMICAL COMPOSITION OF BLOOD, SALIVA AND URINE FROM CYSTIC FIBROSIS HOMOZYGOTES AND HETEROZYGOTES AND HEALTHY CONTROLS

Ceder, O., Bardoń, A., Hellsing, K. and Kollberg, H. (Department of Pediatrics, University Hospital, Umeå, Sweden, Department of Clinical Chemistry, University Hospital, Uppsala, Sweden, Department of Biochemistry, Institute of Sport, Warsaw, Poland and Department of Pediatrics, University of Kuwait, Safat, Kuwait). Effects of sucrose load on the biochemical composition of blood, saliva and urine from cystic fibrosis homozygotes and heterozygotes, and healthy controls. Acta Paediatr Scand, 1983; Suppl. 309:33. A disturbance in the metabolism of carbohydrates has been suggested to be responsible for many of the clinical symptoms in cystic fibrosis. To study this, the biochemical compositions of blood, mixed saliva and urine from homozygotes and heterozygotes for cystic fibrosis and healthy children and adults were compared before and after an oral load of sucrose. In blood, the fasting values of glucose, pyruvate and lactate were similar in the four groups. In saliva, the fasting concentrations of protein, sodium, chloride and potassium were significantly increased for homozygotes. Heterozygote saliva values were intermediate, although not significantly different from those of controls. The urinary concentrations of sodium, chloride, potassium and calcium were also significantly increased in the homozygotes. Again, heterozygotes values were intermediate and did not differ significantly from control values. Sucrose intake caused greater increases in the blood levels of glucose, lactate and pyruvate in the homozygotes and heterozygotes than in their respective control groups. Reversal of the increased concentrations occurred most quickly in the homozygotes and 150 min after sucrose intake the glucose and lactate concentrations were significantly lower in this group than in control children. In saliva significantly lower concentrations of sodium, chloride and potassium were found in the homozygotes and heterozygotes 30 and 90 min after sucrose intake than in the fasting state. Thus, their salivary concentrations of electrolytes became closer to those of controls. In controls sucrose intake caused an increase in the activity of ribonuclease in the saliva while the electrolyte values were unaffected. Limited effects of the sucrose intake were seen on the urinary values in the four groups.