Cytoplasm Of Leukocytes Research Articles

Cytochemical identification of acid deoxyribonuclease in blood cell cytoplasm. 3. Activity of acid deoxyribonuclease in leukocyte cytoplasm of children suffering from acute leukemia

Cytochemical identification of acid deoxyribonuclease in blood cell cytoplasm. 3. Activity of acid deoxyribonuclease in leukocyte cytoplasm of children suffering from acute leukemia

Bactericidal substances in cytoplasm of leucocytes. An attempt to separate lysozyme by gel filtration.

A substance with a bactericidal effect onEscherichia coli was isolated from the cytoplasm of polymorphonuclear leucocytes. It consists of a mixture of different active substances and therefore a gel filtration using s Sephadex G-100 column was used to separate lysozyme from other active substances. The extract was separated by this procedure into five peaks, out of which peak I. possesed a significant bactericidal activity, peak II. and III. had a weak one, whereas the lysozyme activity was present in peak III., IV. and V.

Cytophotometric determination of alkaline phosphatase activity of individual neutrophilic leukocytes with a biochemically calibrated model system.

A model system consisting of polyacrylamide films into which cell sonicate is incorporated was applied to investigate quantitative aspects of the cytochemical azo dye coupling method for alkaline phosphatase activity in neutrophilic leukocytes. The films were assayed in media containing naphthol AS-MX phosphate and 4-aminodiphenylamine diazonium sulfate. Optimal reaction conditions under which proportionality existed between the amount of reaction product and incubation time or enzyme concentration were determined. Since the enzyme activity in the films could also be measured chemically, using disodium phenyl phosphate as a substrate, a direct relation between the cytochemical and biochemical activity could be established. The azo dye coupling method was applied for the quantification of the enzyme activity. Air-dried microscopic preparations of exudate neutrophils were stained and the amount of dye formed in the cytoplasm of individual leukocytes was measured with a cytospectrophotometer based on the two-wavelength principle. By reference to the relation between biochemical and cytochemical activities in the model films, the cytochemically determined enzyme activity could be expressed in biochemical units. Independently, the average alkaline phosphatase activity per cell was determined by direct biochemical assay of the leukocyte suspension. The results showed that about 60% of the biochemically assayed activity in sonicate was demonstrated in the cell preparations by the cytochemical staining method. The discrepancy between the results of the two methods is discussed. The applicability of the model system for elucidation of the relationship between the results of cytochemical and biochemical enzyme determination in cells is stressed.

Iodination of bacteria: a bactericidal mechanism.

Myeloperoxidase, iodide, and H(2)O(2) have a bactericidal effect on Escherichia coli. Myeloperoxidase can be replaced in this system by lactoperoxidase or by a guinea pig leukocyte particulate preparation, H(2)O(2) by an H(2)O(2)-generating system such as glucose and glucose oxidase, and iodide by thyroxine or triiodothyronine. The bactericidal effect was high at pH 5.0 and fell as the pH was increased. Preincubation of myeloperoxidase, iodide, and H(2)O(2) for 30 min before the addition of the bacteria largely prevented the bactericidal effect. Thus, the organisms must be present in the reaction mixture during iodide oxidation for maximum killing, which suggests the involvement of labile intermediates of iodide oxidation rather than the more stable end products of oxidation such as iodine. Iodination of the bacteria by the myeloperoxidase-iodide-H(2)O(2) system was demonstrated chemically and radioautographically. Iodination and the bactericidal effect were similarly affected by changes in experimental conditions in all the parameters tested (effect of preincubation, pH, and inhibitors). Phagocytosis of bacteria by guinea pig leukocytes was associated with the conversion of iodide to a trichloroacetic acid-precipitable form. Iodide was localized radioautographically in the cytoplasm of human leukocytes which contained ingested bacteria. Iodide fixation was not observed in the absence of phagocytosis or in the presence of Tapazole.

Alkaline Phosphatase in Leucocytes by An Azo Dye Method

Alkaline phosphatase activity was demonstrated in the cytoplasm of leucocytes of man, rat and rabbit by means of naphthol AS phosphate as the substrate and fast blue BBN as the coupling agent. The reaction was effected in Tris buffer, 0.2 M at ph 9.1, in 5-10 min at 25°C. Air-dried smears, with or without fixation in 10% neutral formalin, showed activity but those heated 1 min in water at 100°C did not.

Studies on bacteriemia. I. Mechanisms relating to the persistence of bacteriemia in rabbits following the intravenous injection of staphylococci.

During the course of studies on the characteristics of experimental bacteriemia, staphylococci were swiftly cleared from the blood stream of rabbits during the initial 10 to 15 minutes following intravenous injection of microorganisms. A subsequent abrupt decline in the rate of clearance ensued, resulting in a low grade bacteriemia which was demonstrable for many hours. The experiments reported have indicated that this strain of staphylococcus is rapidly phagocyted within the vascular system of rabbits, and that viable staphylococci circulate within the cytoplasm of polymorphonuclear leukocytes. The removal mechanisms contained within the liver and spleen appear to preferentially trap circulating extracellular staphylococci. When most of the circulating staphylococci are contained within leukocytes, splanchnic removal declines or virtually ceases. These observations suggest that viable, intracellular microorganisms are responsible for the persistence of staphylococcal bacteriemia in rabbits following the phase of rapid removal from the blood stream.

Myocarditis and pulmonary arteritis in mice associated with the presence of rickettsia-like bodies in polymorphonuclear leucocytes.

A previously undescribed disease of mice, characterized by acute interstitial myocarditis, and by periarterial pulmonary lesions, is reported. The transmissibility of this disease has been shown, but a large proportion of inoculated mice appears to be resistant. Microorganisms, rickettsial in form but not positively identified as such, are consistently present in the lesions. They are found predominantly in the cytoplasm of polymorphonuclear leucocytes. Filtrates of fecal material from normal mice of the Harvard breeding stock may produce lesions in myocardium and pulmonary arteries indistinguishable from those occurring in this disease. Rickettsia-like bodies are also found in these lesions.

A simple method of preserving the outlines of leucocytes and malarial parasites in giemsa-stained thick blood films

Thick blood films cannot be satisfactorily stained by conventional Giemsa methods until they have been dried for several hours. In thick films which are insufficiently dried diluted Giemsa stain produces a marked dispersion of the cytoplasm of leucocytes and of blood protozoa. Prolonged drying greatly reduces this dispersion, hence the usual recommendation that thick blood films should be dried for several hours before staining. Similar preservation of cytoplasmic contour is obtainable by treating freshly dried films for 1 second with an isotonic solution of methylene blue. This procedure obviates delay from slow drying and is perhaps of interest to those who, because of the delay, hesitate to use thick blood films for routine malarial diagnosis.

The estimation of the bactericidal power of the blood

The survival rate,p, of a measured inoculum ofStaph. aureusin a standard volume of denbrinated blood, is a reliable quantitative measure of the bactericidal power of blood. The number of viable organisms in the inoculum and in the blood-bacterium mixture may be estimated with the necessary accuracy by counts of colonies developing from measured volumes of the fluids let fall on to the surface of solid media. Fildes' agar was the most suitable medium for this surface-viable count, and was selected on the basis of four criteria; of the media tested it yielded the highest counts, and the counts conformed most closely to a Poisson series; and on it the mean colony size was maximum, and the coefficient of variation of colony size was minimum. On this medium, the close conformity of the separate count values to a Poisson series enabled the standard error of the survival rate to be determined from a simplification of the general expression for the standard error of a ratio.The number of colonies growing from a sample of a blood-bacterium mixture may be reduced, not by killing of the individual cocci, but as a result of their aggregation either by agglutinins in the blood, or in the cytoplasm of leucocytes that are phagocytic but not bactericidal. It appears that these mechanisms are unlikely to operate in blood-bacterium mixture containing relatively few organisms; in such mixtures the survival rate is a reflexion of the killing power only.The immunological significance ofphas not been investigated, but the range of values for healthy human adults differs significantly from that for sufferers from chronic staphylococcal infection. Moreover, by the technique employed differences may be detected between individual values ofpthat cannot reasonably be attributed to technical or sampling errors.

Intraleukocytic Bodies in Scarlet Fever

Since Dbhle's first article on Leukocytic Inclusions in Scarlet Fever, a number of workers have investigated the occurrence, significance, and nature of such inclusions but the conclusions derived from these investigations differ very much. D6hle' examined smears of blood fixed by alcohol from 30 cases of scarlet fever and from numerous other diseases, which he does not specify. The smears were stained either with a mixture of Hoppe-Seyler's reagent, water and Michaelis' azure blue, or with an acid solution of orseiles mixed with acid hematoxylin. He also used Pappenheim's stain. By these methods he observed inclusions in the cytoplasm of nearly all neutrophiles almost invariably in scarlet fever. These bodies stained blue with the two first-mentioned stains, and took a pale red with Pappenheim's mixture. Their size and shape varied greatly, some being as large as one-sixth the size of the nucleus, and round or rod-shaped, or oval with one pointed end. The number of bodies per cell varied from one to six. The only cases of scarlet fever in which such bodies did not occur were two examined late in the disease. Only in cases of two other diseases were similar bodies found; viz., two cases of carcinoma and one of pneumonia, in which there was a history of syphilis (the author stating that in this particular instance his slides might have become mixed with those of scarlet fever). D6hle holds that because the bodies stain red with Pappenheim's stain they need not be confused with nuclear fragments; he also finds similar bodies free in the blood. In a more recent article D6hle2 describes spiral forms in the cytoplasm of leukocytes, and also in the blood of two cases of scarlet fever. These he believes are spirochaetes and he names them Spirochaeta scarlatinae. The bodies which he described in the leukocytes he holds to be fragments of the parasites of both diagnostic and prognostic significance. Stimulated by D6hle's work, Kretschmer3 studied some 30 cases of scarlet fever and about 70 controls. At first he used D6hle's method, but soon changed to fixation by methyl alcohol and staining with Manson's stain, because this technic is equally efficient and much simpler. He also tried Giemsa's stain. Kretschmer verified Dbhle's results in regard to scarlet fever, finding bodies in all early cases; they usually disappeared by the 8th-ioth day, occasionally somewhat later. D6hle and Kretschmer both failed to find such bodies in animals; the latter failed to find any relation between the temperature and the occurrence of the bodies. The other diseases in which he found the bodies, but by no means constantly, were diphtheria, tuberculosis, and pneumonia. They were not found in leukocytes from normal persons or in cases of measles. Nicoll and Williams4 used Kretschmer's technic, and also Giemsa's stain over night. In 51 cases of scarlet fever in which the patients had been sick more than eight days the bodies were present in all except six cases. Bodies were also found

NOTE ON THE MULTIPLICATION AND MIGRATION OF NUCLEOLI IN NERVE CELLS OF MAMMALS

The conclusions arrived at from the observations here described are—1. That the nucleoli of the nerve cells described multiply continually, generally by a process of budding, more rarely by an equal division in bulk of a pre‐existing nucleolus.2. That the nucleoli pass out of the nucleus, and in the process their staining reaction changes.3. After passing out of the nucleus the nucleoli become granular in appearance, and increase in size. The increase in size would appear to be due to the lessened density of the contents, rather than to an increase in substance.4. The nucleoli sometimes pass bodily out of the nerve cell (peri‐karyon), and are taken into the cytoplasm of leucocytes or capsular cells. At other times the substance of the nucleoli seems to pass in small portions from the cytoplasm of the nerve cell into that of a capsular cell or a leucocyte.