Fresh Guinea Pig Serum Research Articles

The protective action of some amino-acids against the effect of heat on complement.

1. Glycine, alanine, and several isomers of alanine, DL-glutamic acid and DL-aspartic acid, when added to fresh guinea-pig serum and allowed to stand on the bench for half an hour, will protect the complement of this serum from destruction by heating at 55 and 56° C. for half an hour, but most of the complement activity is destroyed by heating at 57° C. and it is completely destroyed at 58° C. after half an hour.2. Derivatives of glycine do not have any protective effect.3. Various substances of high molecular weight, that might be described as ‘protective colloids’ do not have any protective effect.4. How these amino-acids when added to serum alter the heat-lability of the complement is not understood.

Complement and Opsonic Activities of Fresh Human Sera

Abstract Previous work from this laboratory (1) confirmed earlier observations that there is a thermolabile anti-staphylococcus opsonin in fresh guinea pig serum which operates as a function of its concentration; and that an inactivated antistaphylococcal rabbit serum also behaves as an opsonin. The latter gives a zone effect. It was also noted that there is practically no spontaneous phagocytosis of staphylococci by human leucocytes in the absence of an opsonic agent. In a second paper of the same series dealing with the opsonic power of different mammalian sera (2), the opsonic power of each serum was evaluated with respect to its over-all complement titer, and the conclusion was drawn that “no relationship exists between the hemolytic complementary and opsonic powers of the guinea pig, human, monkey (rhesus), dog, cat and sheep sera toward staphylococci.”

A Dialyzable Component of Complement

When serum of a normal guinea pig is dialyzed against a normal saline solution with a cellophane tube the hemolytic activity of the solution decreases rapidly. A prolonged dialysis will destroy almost all of the original activity. We have now found that this loss of activity is due to the loss of one of the components of complement which passes through the membrane into the saline solution, and that the original activity may be almost completely restored by the addition of the dialyzable to the non-dialyzable fraction. A typical experiment of the dialysis is as follows: 10 cc of fresh guinea pig serum containing 80 units of complement per cc were poured into a cellophane tube both ends of which were closed with rubber stoppers and tightened with rubber bands. The sealed tube was then suspended in a graduated cylinder containing 100 cc of a cold normal saline solution. The dialysis was allowed to take place at about 5°C. After 24 hours, the bag was opened and a half cubic-centimeter of serum was pipetted out, and it was found to contain 50 units per cc. The rest of the serum was again dialyzed with another 100 cc portion of a normal saline solution. After 24 hours another half cc of the serum was taken out and its hemolytic activity was found to be 20 units per cc. The rest of the serum was again dialyzed with a fresh volume (100 cc) of the saline solution. Two days later the bag was opened and the dialyzed serum now contained less than 10 units per cc. The total volume of serum after dialysis was about 9 cc. Therefore the decrease of activity was not due to the dilution of serum. A control sample of the same guinea pig serum was stored without dialysis in another cellophane tube and kept at about 5°C. Its complement titer dropped from 80 to 60 only in 4 days. It was of interest to ascertain whether the loss of activity during dialysis is due to the separation of the components of complement or to some chemical reaction such as oxidation. We have found that the original complementary activity may be restored by combining the dialyzed serum and the dialyzate. This was done as follows: To 0.5 cc of the dialyzed serum diluted 30-fold with 0.35% NaCl were added varying amounts of dialyzate, and enough normal saline solution to make a total volume of 2 cc. Five-tenths cc of hemolysin (2 units/cc) and 0.5 cc of 2% sheep cells were then added. After 30 minutes at 37°C readings were made. The results of a representative experiment are presented in Table I.

The Influence of Bovine Serum on Brucella Infection in Guinea Pigs

Antiserums have long been used in the field to combat brucella infections in large animals. As pointed out by Gwatkin,1 brucella antiserum has not been used many times under controlled conditions. Inasmuch as he has given a complete bibliography, the literature will not be reviewed here. Gwatkin1 used guinea pigs and injected them with single injections, 1-10 cc, of fresh serum from cows infected with Br. abortus. From these investigations he concluded that Br. abortus infection in guinea pigs was delayed. Single injections, .5 to 1 ? 2 cc. of hyper-immune guinea pig serum produced a similar delay in the onset of infection as indicated by the agglutination titre; single injections of 5 cc. of fresh hyperimmune guinea pig serum produced a like result. Gwatkin's next report2 deals with the toxicity of fresh cow serum for guinea pigs when given intra-abdominally. He concludes that fresh cow serum cannot be given to guinea pigs without danger. Heating the serum for 30 minutes at 56 renders it innocuous for these animals. The writer3 had under observation a herd of dairy cattle. It was found that the bactericidal power of the whole blood of these cows for killing brucella varied between less than 100 organisms to 80,000 organisms per cc. In an endeavor to determine whether normal serum showing a high bactericidal power would, when injected into guinea pigs, enable them to resist brucella infection the following trials were made:

A METHOD OF FACILITATING THE SERUM DIAGNOSIS OF SYPHILIS UNDER WAR CONDITIONS

The necessity for fresh guinea-pig serum and fresh sheep corpuscles in the original Wassermann technic renders that test impracticable unless these two factors are obtainable in the laboratory. I have already introduced a procedure in which the test is made possible without the use of fresh sheep corpuscles, and this antihuman hemolytic method has found wide adoption, notably in the laboratories of the United States Army and Navy. This test, however, still requires the use of fresh guinea-pig serum as complement. Considerable apprehension is being felt at present with regard to the availability of guinea-pigs and sheep to supply the necessary material for carrying out the serodiagnostic test for syphilis according to the methods hitherto in use, and a method which renders the test independent of these resources should be welcome at this time, when economy of labor, expense and time is particularly urgent. That such a method is possible,

A Contribution to the Action of Amidoacids, Peptids and Proteins on Hemolysis by Cobravenom

A Contribution to the Action of Amidoacids, Peptids and Proteins on Hemolysis by Cobravenom

Natural Hemolysins in Normal Horse Serum

During the study of the complement-fixation test for glanders in horses, our attention was attracted by the question of a natural hemolysin for the corpuscles of the sheep and the ox. A search of the literature has failed to give us the desired information. Morgenroth and Sachs1 state is worthy of note that in almost every horse serum which we examined for the purpose, we found a considerable amount of amboceptor for guinea-pig blood. The amboceptor was characterized by a particularly high degree of thermolability, being invariably destroyed by heating to 55 degrees C. A complement for the same is very often absent, and even when present it is only on the addition of considerable amounts of fresh guinea-pig serum that this amboceptor becomes manifest. Sachs,2 in his table compiled from the literature, mentions as doubtful the presence of natural amboceptors for the erythrocytes of guinea-pigs and rabbits, and an absence of the same for the erythrocytes of other vertebrates.

Societies and Academies

LONDON. Royal Society, January 28.—Sir William Crooks, president, in the chair.—W. W. C. Topley: The influence of salt-concentration on haemolysis. The method employed in this investigation has consisted in varying the percentage of sodium chloride present in a mixture of sheep's red cells, hæmolytic anti-body and complement. The hæmolytic anti-body was obtained from an immunised rabbit, and fresh guinea-pig serum, previously absorbed with sheep's red cells at 0° C, furnished the complement. Where hypotonic percentages of sodium chloride have been employed, the solutions have been made in 7.8 per cent. saccharose solution. The results have confirmed the observations of other workers that, when the percentage of salt is increased beyond the normal limit, hæmolysis is inhibited, owing to the failure of union between the complement and the red-cell anti-body complex, and that, if the amount of anti-body present in the mixture be increased, the effect of the increased salt concentration may to a certain extent be overcome. It has further been shown that, if the salt concentration be decreased below the normal limits, less and less anti-body becomes necessary in order to bring about the union of red cells and complement, and that, in the almost entire absence of electrolytes, guinea-pigs' complement can hæmolyse sheep's red cells without the intervention of hæmolvtic anti-body.—G. Smith: The life-cycle of Cladocera, with remarks on the physiology of growth and reproduction in Crustacea. As the result of breeding experiments, it is shown that the effects of isolation, crowding, and temperature are the same for Daphnia pulex as previously reported for Moina rectirostris, viz., isolation combined with high temperature completely inhibits the production of sexual individuals, reproduction being entirely parthenogenetic. Crowding, combined with low temperature, results sooner or later in the production of males and ephippial females.—T. Goodey: Investigations on protozoa in relation to the factor limiting bacterial activity in soil. Two different lots of soil were taken and inoculated with cultures of protozoa originally obtained from soil, in order to determine whether the added protozoa would cause a decrease in the numbers of bacteria in the soil. One soil was free from protozoa to begin with, having been bottled since 1846, whilst the other was freed from protozoa by partial sterilisation. Separate lots of each soil were inoculated with cultures of ciliates, amoebae, and flagellates, and suitable control sampleswere set up. Periodical bacterial counts were made inorder to ascertain if the protozoa were exerting a limiting action on the soil bacteria, and these counts were continued over a period of about eighteen months. The general conclusion drawn from the investigations is that ciliates, amoebae, and flagellates do not act as the factor limiting bacterial activity in soil.—S. Hatta: The mesodermic origin and fate of the so-called mesectoderm in petromyzon. The name mesectoderm has given to a loose aggregation of mesenchymatous tissue, in some places assuming the character of an epithelium intervening between the myotomes and the ectoderm in the head and branchial region of the embryo of petromyzon. As the name implies, it has been confidently asserted that this tissue is derived from the ectoderm. In this paper, however, it is shown that this tissue originates from the ventral edge of the myotome and corresponds to the ventral extension of the myotome in the trunk region which grows downwards towards the mid-ventral line outside the splanchnocœle and gives rise to the ventral muscles of the trunk.—Prof. J. C. Bose: The influence of homodromous and heterodromous electric currents on transmission of excitation in plant and animal. The action of an electric current in inducing variation of conductivity may be enunciated under the following laws, which are equally applicable to the conducting tissue of the plant and the nerve of the animal:—(1) The passage of a current induces a variation of conductivity, the effect depending on the direction and intensity of current. (2) Under feeble intensity, heterodromous current enhances and homodromous current depresses the conduction of excitation. (3) The after-effect of a feeble current is a transient conductivity-variation, the sign of which is opposite to that induced during the continuation of current. (4) The normal conductivity-variation undergoes a reversal under a strength of current above the critical value. The heterodromous current then induces a depression while the homodromous current induces an enhancement of conductivity.

Studies on the Berkefeld-Filtration of Complement

In the Berkefeld filtration of a fresh guinea-pig serum, adsorption on the filter surface takes place, and the serum passes unaltered, if the saturation of the adsorbing surface has become complete.This adsorption involves most probably first the albumins, which process then secondly influences the filtrability of the globulins. It is also shown that the physical conditions of the serum as regards salt concentration and dilution as well as H.-concentration modify the effect of filtration to a large extent.

The Formation of Antibodies in Rats Fed on Pure Vegetable Proteins (Osbornemendel Stunting Food)

Osborne and Mendel have made the important discovery that by feeding white rats certain pure vegetable proteins normal growth is arrested without disturbance of the general health, the animals resuming growth on the resumption of ordinary food or when certain amino acids are furnished.' As I was engaged in a study of the formation of antibodies under different conditions, it occurred to me that it would be of interest to try to determine whether or not the formation of antibodies proceeds in the usual way in rats whose growth is arrested by this means. Accordingly, the amount of specific lysin for sheep corpuscles, produced in normal and stunted animals under comparable conditions, has been determined. The blood-serum of white rats normally causes lysis of sheep corpuscles. This action is dependent on an amboceptor-complement mechanism, because on being heated at 56 C. for thirty minutes the serum loses its lytic power, which is restored, however, on the addition of complement in the form of fresh guinea-pig serum, in quantities that in themselves do not cause laking of sheep corpuscles. The amount of amboceptor in normal rat serum varies, but usually no lytic action is obtained in dilutions of the serum above 1 to 96. When white rats are

THE NOGUCHI SERUM REACTION FOR SYPHILIS AS AN AID TO DIAGNOIS IN EYE LESIONS

The serum diagnosis of syphilis as given to the world by Wassermann, Neisser and Bruck1in 1906, has now become a well-established diagnostic method, generally used in all medical centers, and has proved of value in all fields of medicine. The Wassermann method is an adaptation of the Bordet-Gengou phenomenon of complement deviation, in which the hemolytic system consists of a suspension of washed red blood cells of the sheep, hemolytic amboceptor made by immunizing rabbits to sheep's red blood cells, and as complement fresh guinea-pig serum. For syphilitic antibody the serum from suspected patients is used, and for antigen a watery extract of a liver of congenital syphilis known to be rich inSpirochaeta pallida. It has been pointed out that there is one factor of error in this method due to the fact that there is in human serum a variable amount of natural antisheep amboceptor which