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Specific extraction of antigen in vivo by extracorporeal circulation over antibody immobilized in collodion-charcoal.

A novel extracorporeal immunoadsorbent is described in which anti-bovine serum albumin (anti-BSA) was entrapped in collodion membranes adherent to activated charcoal particles. This immunoadsorbent was capable of specifically removing circulating BSA in vitro and in vivo in an extracorporeal system in dogs. In preparation of the immunoadsorbent, up to 81.2% of added anti-BSA was retained on collodion-charcoal. In vitro circulation studies demonstrated that anti-BSA collodion-charcoal removed 812% more 125I-BSA than control colloidon-charcoal. For in vivo studies, an extracorporeal circulation system was established and arterial blood was passaged through a continuous flow celltrifuge wherein plasma which was separated from formed elements of the blood was circulated over anti-BSA and control rabbit gamma-globulin collodion-charcoal. 125I-BSA was passively infused into mongrel dogs, and plasma was then circulated over extracorpeal immunoadsorbents for 2 hr. Results showed up to 896% greater uptake of circulating 125I-BSA on the charcoal containing immobilized anti-BSA compared to control charcoal. There was no evidence of release of anti-BSA from the immunoadsorbent since 125I-anti-BSA cpm on the charcoal before and after the experiments were unchanged. In addition, there were no significant alterations in hematocrit, leukocyte counts, serum sodium, potassium, calcium, magnesium, or creatinine levels before and after in vivo procedures. These data suggest that this immunoadsorbent consisting of anti-BSA immobilized in collodion membranes adherent to charcoal particles may specifically withdraw circulating antigens in vivo with minimal release of entrapped antibodies and no significant alteration in the host hematologic and biochemical status.

Animals

Specific removal of antibody by extracorporeal circulation over antigen immobilized in collodion-charcoal.

Studies were undertaken to determine whether BSA immobilized in collodion membranes adherent to activated charcoal particles, would be capable of specifically removing circulating BSA antibody in vitro and in vivo in an extracorporeal system in dogs. Up to 59-8 mg of BSA were retained in collodion membranes adherent to small particles. In vitro studies demonstrated that immobilized BSA could specifically reduce BSA binding activity from circulating antisera. For in vivo studies, an extracorporeal circulation system was established and arterial blood was circualted through a continuous flow celltrifuge in which plasma was separated from formed elements of the blood. Only plasma was circulated over the BSA collodion-charcoal immunoadsorbent. Anti-BSA and anti-HSA atibodies were passively infused into dogs and, after an equlibration period of 12 or 15 min, plasma was passed over the BSA collodion-charcoal immunoadsorben. Plasma exhibited a sharp reduction in BSA binding over the next 30-60 min with only slight reduction in anti-HSA binding the same period. Dogs, actively immunized to BSA and HSA, were also treated by extracorporeal plasma perfusion over BSA collodion-charcoal. A specific decline in BSA binding of sera, was again observed with minimal changes in HSA binding. A post-perfusion rebound of BSA binding was observed which reached pre-perfusion levels after 6-8 days. A second treatment during the rebound period also resulted in a specific decline in BSA binding with a similar pattern of antibody rebound. There were no significant changes in I-labelled BSA recorded on the charcoal before and after in vivo procedures and no signifcant alterations in haematocrit, serum sodium, potassium, calcium, magnesium or creatinine levels before and after the procedures. These data suggests that antigen-coated charcoal may specifically withdraw circulating antibodies in vivo with minimal release of the entrapped antigen and little alteration in the host's haematological and biochemical status.

Animals

[Collodion baby. Clinical course based on 29 cases].

Study of 29 cases of collodion baby and 169 cases previously published led to several conclusions. In the short-term, the child is threatened by pulmonary infections. The increased skin permeability may result in the loss of water or intoxication by cutaneous absorption of a substance applied to the skin. In the long-term the condition progresses to one of a congenital disturbances in keratinisation. In 2/3 of cases, the problem is that of congenital non-bullous ichthyosiform erythrodermia. Profression to lamellar ichthyosis, ichthyosis vulgaris, circumflex linear ichthyosos and Conradi's syndrome is established. Ichthyosis related to sex in our cases did not seem to be a feature of the collodion baby, the opinion in the literature being divergent. Congenital bullous ichthyosiform erythrodermia never begins as collodion baby syndrome.

Adolescent

The keratinization disorder in collodion babies evolving into lamellar ichthyosis. Its possible relevance for determining the primary defect in lamellar ichthyosis.

Two collodion baby girls with disorder evolving into lamellar ichthyosis were followed by light and electron microscopy. Light microscopically, the neonatal colloidion skin was characterized by a thick compact stratum corneum which was PAS positive in its upper two thirds, by a thin stratum granulosum and by a non-acanthotic stratum spinosum with normal mitotic activity. Electron microscopically, the upper stratum corneum appeared pathological, whereas the lower part was normal except for some minor parakeratosis. The main alterations in the underlying stratum granulosum were diminished tonofibrils and keratohyalin. Biopsy specimens taken at the age of 2 weeks were typical for lamellar ichthyosis and showed hyperkeratosis with focal parakeratosis, a thickened stratum granulosum in which the cellular content of keratohyalin and tonofibrils was moderately diminished, and acanthosis with increased mitotic activity. It appears that the ultrastructural changes of the stratum granulosum, seen in lamellar ichthyosis, are already present in the collodion skin of the newborn, at a time when the epidermis does not yet show an increase in mitotic activity.

Female

[Correlation of structural and ultrastructural modifications of the renal cortex with water, sodium and protein excretion in rats rendered hypertensive by perinephritic constriction by collodion].

Water, sodium and proteins renal excretion in with collodion Page's method hypertensive rats is compared with that of same Wistar strained control rats. Blood pressure of treated animals (n = 16) is 193.1 Hg mm (138 in control rats). Urinary flow increases from 19.5 to 33.7 ml/24 h (+ 72.8%), sodium excretion from 29.6 to 37.5 mg/24 h (+ 26.7%) and total proteins excretion from 23.5 to 63 mg/24 h (+ 169%. This data are correlated with the renal cortex morphological changes with photon and electron microscope. Severe damages are seen in Bowman's capsule and in glomerular copruscules, especially at the epithelial layer level. Important proteic pools occur within tubular lumen. Proximal tubular epithelium seems normal. On the other hand, distal tubular epithelium seriously scales. So, important water, sodium and proteins excretion increase in our experimental hypertension model can be explained. It is also discussed about functional and structural modifications analogy in our model's rats and in spontaneously hypertensive rats (SHB).

Blood Pressure

A technique for the freeze-fracture of tissue culture.

Cultures of rat cerebellum grown on glass coverslips coated first with collodion and then collagen are fixed in glutaraldehyde to which ruthenium red has been added. After fixation, the collodion comes off the glass coverslips and the 3 layers: collodion-collagen-culture are handled as one piece. Following immersion in glycerol, selected areas are cut from the piece; the excised areas are a little smaller than those of the specimen carriers used in a Balzers apparatus. The ruthenium red, which stains all cells and processes, is a good indicator of specimen thickness. The explant which consists of several cell layers is redder than the peripheral monolayer of outgrowth. The square areas cut from either the explant or the outgrowth can be matched in thickness according to their shade of red. In frozen specimens, the red colour distinguishes the culture layer from the collodion-collagen support under it and from the white frost on top. During fracturing, the appearance of pink flakes on the edge of the knife indicates that the culture layer has been cleaved. The growth of cells on collodion-collagen and the fracture of the cultures with a knife, eliminate the restrictions inherent in the use of frozen 'metallic sandwiches'.

Animals

Attenuation of canine nephrotoxic glomerulonephritis with an extracorporeal immunoadsorbent.

A novel immunoadsorbent is described in which soluble canine glomerular basement membrane (GBM) antigen was immobilized in collodion membranes adherent to small charcoal particles. As much as 80% of the added GBM antigen was retained in the collodion-charcoal. The binding of GBM antibody to immobilized GBM antigen in vitro and in vivo was demonstrated. For in vivo studies, an extracorporeal circulation system was established, and mongrel dogs were then injected intravenously with GBM collodion-charcoal for 4 h. Renal tissue was examined by fluorescence and light microscopy for the deposition of heterologous gamma globulin and glomerular inflammation before and after the extracorporeal procedure. Results showed less deposition of heterologous IgG and less inflammation in experimental dog kidneys than in control kidneys. There was no evidence of release of 131I-labeled GBM from the immunoadsorbent at the conclusion of perfusion experiments and no significant biochemical or hematologic alterations. Finally, extracorporeal immunoadsorbent treatment was capable of attenuating the early heterologous phase of passive canine nephrotoxic nephritis and may have a potential role in the therapy of anti-GBM-mediated nephropathy.

Animals

Thermoosmosis through charged membranes.

Thermoosmosis through oxidized collodion and collodion-sulfonated polystyrene interpolyrene interpolymer membranes has been observed in KCl solutions of various concentrations. The effective temperature difference acting for thermoosmosis was determined by measuring the thermal membrane potential appearing on both sides of membrane. It was found that the velocity of thermoosmosis is proportional to the effective temperature difference and the proprtionality constant (themoosmotic coefficient) is a function of electrolyte concentration. The dependence of the thermoosmotic coefficient of charged membranes on the electrolyte concentration is found to have a characteristic feature.

Collodion

[In vitro evaluation of adsorbents used in haemoperfusion (author's transl)].

The properties of six adsorbents for Haemoperfusion were investigated in vitro: Amberlite XAD-2; Amberlite XAD-4; Haemocol; collodion coated charcoal; albumine-collodion encapsulated charcoal and activated carbon fibers. Carbromal and diazepam are most quickly removed from aqueous solutions by Amberlite XAD-2 and XAD-4. Of the charcoal containing materials, Haemocol eliminates most rapidly carbromal and diazepam; while activated carbon fibers remove phenobarbital faster. When heparinized blood is perfused, a remarkable reduction in platelet count is observed. The drop is most pronounced with Amberlite XAD-2 and XAD-4, but even when using charcoal preparations the platelet depletion can reach 10% of the original number over 5 hours. Haemolysis is most pronounced with Amberlite XAD-2, XAD-4 and Haemocol. As the efficiency of the adsorbents towards the various drugs differ and as their adverse effects also vary, the decision for a particular material will have to be made according to the individual case.

Adsorption