THE ETIOLOGY OF VERRUGA PERUANA

A saline suspension of a subcutaneous nodule excised from a verruga patient, and kept in the refrigerator for 14 days, on inoculation into two young Macacus rhesus monkeys (Nos. 33 and 34) induced irregular febrile reactions and enlargement of the lymph glands, and in one instance a subcutaneous nodule arose, independently of direct inoculation, on the tail. A microorganism has been isolated from the blood of both animals, and from the experimental nodule, which in pathogenic properties and in cultural and morphological characteristics is indistinguishable from the strain of Bartonella bacilliformis isolated from a case of Oroya fever. The spontaneous skin lesion of Monkey 34 and the subcutaneous nodules induced by intradermal inoculation of cultures of the microorganism were histologically typical of experimental verruga lesions in monkeys and identical with the skin lesions induced in monkeys by Bartonella bacilliformis. The organism, like Bartonella bacilliformis, is an intracellular parasite, being found in the cytoplasm of the proliferating endothelial cells of the lesions and in the erythrocytes of the blood. The same variations in the manifestations of disease which have been noted in experimental infection with Bartmetta bacilliformis were observed in the experimental verruga infection. In the second passage (Monkey 41) the infection induced by local inoculation of cultures was severe both locally and constitutionally and was accompanied by marked anemia. The organisms were found in the red cells in large numbers. In the third passage the systemic infection was less severe, but the local lesions were more striking. Detection of the parasites in the blood is far more certain by the cultural method than by microscopic examination, the latter procedure being successful only in rather severe infections. The result of blood culture is therefore the decisive method in the final diagnosis of the disease. Preliminary serological study shows that the organism isolated in the present instance from the skin lesion of a verruga patient and that previously obtained from the blood of a case of Oroya fever belong to the same serological group. The data obtained justify the conclusion that verruga peruana is caused by Bartonella bacilliformis. They also definitely establish the fact that the inoculation of blood or sanguineous exudate from lesions of verruga peruana is capable of inducing in susceptible individuals a severe febrile systemic infection, such as that to which Carrion succumbed. The designation "Carrion''s disease" is therefore the appropriate one for both forms of the infection. Bartonella bacilliformis may be regarded as a bacterium, since it has the essential features of that group of microorganisms.     

ETIOLOGY OF OROYA FEVER : VIII. EXPERIMENTS ON CROSS-IMMUNITY BETWEEN OROYA FEVER AND VERRUGA PERUANA.

Nine monkeys (Macacus rhesus) and a chimpanzee which had recently recovered from an infection with the Oroya strain of Bartonella bacilliformis were tested for immunity against the verruga strain of Bartonella bacilliformis as well as against the homologous strain. Complete immunity to both strains was demonstrated. The result establishes the identity of the strains and is in agreement with the result of comparative serological study. The criteria of recovery include not only the subsidence of febrile reactions and local lesions but also a negative result of cultural tests of blood and lymphatic gland tissue. Recovery may occur as early as 1 month after inoculation, but in most instances a period of 2 to 5 months is required for the completion of convalescence.     

STUDIES ON X-RAY EFFECTS : XIV. THE EFFECT OF X-RAY ON THE DIVISION RATE OF PARAMECIUM.

Two races of paramecium were submitted for varying lengths of time to x-radiation and a large number of individuals were observed to determine the effect on the rate of division. The division rate of both races suffered a slight initial depression lasting for 2 to 5 days following the exposure. This depression is followed by complete recovery. Within rather wide limits the length of the exposure has, in these experiments, made no appreciable difference. Apparently the maximum effect of the x-rays is produced by relatively short exposures. Continued radiation produces little further change until exposures of 3 and 4 hours are used, when precisely the opposite results are obtained from those obtained with shorter exposures. Doses repeated at various intervals have in general failed to interfere more markedly with the division rate than a single dose. Repeated radiation causes the cells to become slightly swollen without apparent interference with their viability.     

X射线对血液流变性质的影响

本文对３８例从事Ｘ射线工作者进行血液流变检测，结果血浆比粘度２０～２９岁对照组高于受检组，４０岁以上受检组高于对照组。血沉３０～３９岁和４０岁以上两组受检组高于对照组。提示年龄愈大，接触Ｘ线时间愈长，血浆比粘度增高，血沉增快。     

ETIOLOGY OF OROYA FEVER : IX. BACTERIUM PERUVIANUM,N. SP., A SECONDARY INVADER OF THE LESIONS OF VERRUGA PERUANA.

A minute, pleomorphic, motile, Gram-negative bacterium has been isolated from two specimens of nodular tissue from human verruga. In films and sections of the original tissues the organism in question is difficult to distinguish from Bartonella bacilliformis, with which it was associated, and even in pure culture it has a number of properties in common with that parasite. No sugars are fermented by it, it is an obligate aerobe, the optimum temperature for its growth is 25°C., and it has two to four spiral flagella attached to one end of the body. It is, however, readily cultivated on any ordinary culture medium. Broth cultures contain much mucin, but no hydrogen sulfide is formed. Coagulated serum is liquefied by its growth, and the red corpuscles in a blood agar plate are hemolyzed. Rabbits, guinea pigs, rats, and mice develop acute, fatal septicemia as a result of intravenous or intratesticular inoculation of young cultures. The liver is characteristically affected and shows a general parenchymatous degeneration and necrosis; the entire gastrointestinal tract is intensely congested, and numerous hemorrhagic areas are present; the spleen, dark and soft, is rarely much enlarged; the kidneys are swollen and congested; the adrenals are much swollen and intensely red; the lungs are sometimes congested but otherwise normal. In the case of intratesticular inoculation the scrotum and testicle both undergo rapid gangrene. In monkeys no septicemia has been observed, but a violent local reaction—swelling, congestion, sometimes necrosis—follows intradermal inoculation. Since no microorganism corresponding in character with this one has previously been described, it is regarded as a new species, and because of its presence in material obtained from Peru it has been given the name Bacterium peruvianum. The significance of the association of B. peruvianum with Bartonella bacilliformis deserves further investigation; it is not impossible that the two organisms are introduced into the human body by the same blood-sucking insect.     

THE EFFECT OF X-RAY ON THE SKIN OF VITALLY STAINED WHITE MICE : ESTABLISHMENT OF AN X-RAY UNIT FOR THE SKIN OF THE MOUSE.

1. The time interval between radiation and the occurrence of epilation is shorter in mice injected with trypan blue than in normal animals. 2. An x-ray unit defined as that causing total spontaneous epilation on the skin of the mouse is suggested. It corresponds to four to five human erythema doses.     

高能X线照射对肺癌转移相关基因的影响

目的探讨高能X线照射对肺癌转移相关基因的影响。方法建立Lewis肺癌模型，将荷Lewis肺癌的C57BL／6雄性小鼠35只随机分为照射组（n=18）和对照组（n=17），照射组接受30Gy的X线肿瘤局部照射，对照组不接受照射，2周后采用免疫组化法检测瘤组织中E-钙黏素（E—Cad）蛋白和基质金属蛋白酶-9（MMP-9）蛋白的表达。结果与对照组比较，照射组MMP-9蛋白表达水平显著降低，E—Cad蛋白表达水平显著升高，差异均有显著性（t=9．53、20．68，P〈0．01）。结论X线照射可能通过抑制肿瘤组织内MMP-9蛋白表达和促进E—Cad蛋白表达阻止肿瘤细胞的侵袭和转移。     

ON THE SIGNIFICANCE OF THE SUBMILIARY MYOCARDIAL NODULES OF ASCHOFF IN RHEUMATIC FEVER

1. In rheumatic myocarditis, foci, termed submiliary nodules of Aschoff, are present which are characteristic of the rheumatic infection. 2. They are most frequently found in the walls of the left ventricle, the auricles usually escaping. 3. The nodules were found in three cases of chorea without joint manifestations, proving the close relation of this condition to rheumatism. 4. They were absent in fourteen cases of subacute bacterial endocarditis due to Streptococcus mitis. 5. They were not found in infections of the endocardium with the gonococcus, staphylococcus, streptococcus, or pneumococcus. 6. Even in the absence of a rheumatic history we believe, in accordance with Fraenkel, that the presence of Aschoff bodies signifies a previous rheumatic infection. 7. Aschoff bodies are not always found in rheumatic carditis, where the infection antedates death by a long period, but the healed remains, represented by sclerotic patches ("Schwielen"), are present. 8. We suggest that the cases of arthritis characterized by the presence of the submiliary nodules of Aschoff in the myocardium be placed in one group and called for the time being "rheumatism"; and the cases with articular manifestations, yielding positive bacteriological findings and no Aschoff bodies, should be classified according to the infecting microörganisms concerned, and not as rheumatism.     

STUDIES ON X-RAY EFFECTS : V. EFFECT OF SMALL DOSES OF X-RAYS OF LOW PENETRATION ON THE LYMPHOID TISSUE OF MICE.

A dose of x-rays governed by the following factors induces a stimulation of lymphoid tissue in mice: spark-gap ⅞ inch, milliamperage 25, distance 8 inches, time of exposure 10 minutes. Within 4 days after this dose there appeared an abnormally large number of mitotic figures in the lymphoid tissue of spleen and lymph glands, indicating an acceleration of the proliferative activity of the tissue.     

STUDIES ON X-RAY EFFECTS : VI. EFFECT OF THE CELLULAR REACTION INDUCED BY X-RAYS ON CANCER GRAFTS.

Small areas of the skin in the groin of mice were subjected to an erythema dose of x-rays and a week later a cancer graft was inoculated intracutaneously into the area and at the same time a like graft was inoculated in the same manner in the opposite groin protected from x-rays. The graft in the x-rayed area showed a low percentage of takes, while that in the normal skin gave the usual high percentage When the graft was introduced into the subcutaneous tissues it grew equally as well in the x-rayed area as in the protected area. Histological examination shows the skin layers, a few days after x-ray treatment, to be markedly infiltrated with round cells of the lymphoid type. The reaction did not extend deeper than the skin layers. It is suggested that this local lymphoid reaction induced by the x-rays controls the graft made into the skin, while its absence in deeper tissues accounts for the growth of the grafts more deeply implanted.     

低分割模式腹部X线照射对大鼠肾放射生物学效应的影响

目的探讨低分割模式腹部X线照射对大鼠肾放射生物学效应的影响,为低分割模式在腹部照射的临床应用提供参考依据。方法 125只Wistar大鼠随机分为对照组、4 Gy组、6 Gy组、8 Gy组、12 Gy组,每组各25只。各照射组分别接受相应分割剂量的分次照射,测定大鼠放射结束后第2、4、6、8及10周肾脏系数、血尿素氮（BUN）、肌酐（Scr）的变化。苏木精-伊红染色观察大鼠肾脏的形态学变化,免疫组织化学法检测凋亡蛋白Bcl-2及Bax的表达。结果各照射组均可观察到不同程度肾脏体积缩小,皮质比例减小,肾小球毛细血管袢增厚,血管腔扩大,肾小管上皮细胞间隙增大。各组间肾脏系数、Scr、BUN比较,差异有显著性（F=11.833～781.972,P均〈0.001）;不同时间点肾脏系数、Scr、BUN差异有显著性（F=20.857～264.692,P均〈0.001）;不同剂量和不同时间点之间的交互效应比较,差异有显著性（F=2.139～27.550,P均〈0.001）。各组间Bcl-2及Bax表达差异有显著性（F=211.607、116.577,P均〈0.001）,不同时间点Bcl-2及Bax表达差异有显著性（F=54.083、68.749,P均〈0.001）,不同剂量和不同时间对Bcl-2和Bax的交互效应差异均有统计学意义（F=5.032、4.385,P均〈0.001）。结论腹部低分割模式照射可造成大鼠肾脏的放射性损伤,损伤程度呈剂量和时间依赖性,其机制可能与促凋亡蛋白Bax高表达、抑制凋亡蛋白Bcl-2低表达引起细胞凋亡有关。     

THE EFFECT OF PULMONARY CONGESTION ON THE ON THE VENTILATION OF THE LUNGS

1. A method is described for producing pulmonary congestion, together with what may be termed a differential spirometer method for studying lung ventilation. 2. The method utilized permits an approximately accurate prediction of degrees of pulmonary edema in the living animal, and suggests avenues of approach for the very difficult problems of pulmonary capillary pressure. 3. It is shown that intravascular blood can encroach markedly upon the pulmonary air space. Although the methods used in these animal experiments do not resemble vital capacity measurements in man, their result is so definite that their applicability to clinical conditions may be considered. 4. The similarity between the experiments described and certain conditions of cardiac decompensation, of which mitral stenosis is the best example, is pointed out.     

STUDIES ON X-RAY EFFECTS : VII. EFFECT OF SMALL DOSES OF X-RAYS OF LOW PENETRATION ON THE RESISTANCE OF MICE TO TRANSPLANTED CANCER.

A relatively increased degree of resistance in mice to a certain strain of transplantable cancer was demonstrated after treatment of animals with small doses of x-rays capable of stimulating lymphoid tissue. The refractory state induced was determinable 3 to 7 days after the dose of x-rays was given.     

EFFECT OF THE INJECTION OF BILE ON THE CIRCULATION

1. We have confirmed previous work which shows that the injection of bile increases the tone of the vagus nerve, and that this action can be abolished after the administration of atropine. 2. We have found that the amount of bile salts in a lethal dose of pig''s bile for dog''s will, if inejcted alone, produce neither a fall in blood pressure nor a slowing in rate. 3. We have found that the amount of pigment in a lethal dose of the bile will, if injected alone, cause death with slowing of the heart and lowering of blood pressure. 4. We have found that the bile pigment in combination with calcium or sodium is less toxic than uncombined pigment. 5. We have found that in experimentally produced jaundice the calcium content of the blood is increased, while that of the liver, muscle and brain are decreased. 6. We are of the opinion that increase in calcium in the blood is a protective mechanism against the circulating pigments of obstructive jaundice.     

THE EFFECT OF HEAT ON ANTIBODIES

It is possible by means of curves to depict graphically the behavior of the various antibodies under various conditions. Logarithmic curves based on the data presented in the tables are submitted in Text-fig. 1. In general it is evident that antibody destruction goes on gradually as the temperature is increased. Thus 65°C. for 20 minutes diminishes the activity of all the antibodies with the exception of red cell agglutinin, and in this case although the final titer was the same evidently some of the antibody was inactivated, since the reaction was weaker in the higher dilutions. It can, then, be said that 65°C. for 20 minutes appreciably affects the activity of all the antibodies tested. When the temperature is increased to 70°C. more marked differences are apparent. Here both types of the bacterial agglutinin and the precipitin are fairly stable when compared with hemolysin and red cell agglutinin. In both instances there is a sharp decline in the activity of the antibody. 75°C., however, is even a more critical temperature since at this point the somatic bacterial agglutinin and the precipitin are completely inactivated. The hemolysin and hemagglutinin behave alike. The flagellar agglutinin is the most resistant of the group to this temperature. When the temperature is increased to 80°C. the red cell agglutinin is completely inactivated, but sufficient hemolysin still remains to give a slight reaction at the lowest dilution. A further increase to 85°C. completely destroyed the hemolysin but left a definite amount of flagellar agglutinin; in fact, 90°C. for 20 minutes did not completely destroy this substance, since well defined clumps in the lower serum dilutions could be detected on microscopic examination. In this respect, then, the observations of Beyer and Reagh and Orcutt that there is a well defined difference between the two agglutinins for the hog cholera bacillus have been confirmed. However, each substance tested, with perhaps two exceptions, differs in its behavior to heat. It is of interest to point out the similarities in the reaction of somatic agglutinin and precipitin. Both are diminished when heated to 65°C.; 70°C. further affects the agglutinin, but not the precipitin; 75°C. completely inactivates both. The assumption that the substances are apparently destroyed when they cease to react visibly with their respective antigens seems well founded since they cannot be reactivated with normal serum and no longer react to divert complement when combined in a hemolytic system. It might be of interest to mention briefly other experiments in which the temperature was kept constant and the time varied. Thus temperatures of 50–55°C. and 60°C. maintained for 8 hours had no effect on antibody. 60°C. for 4 days failed to alter materially the flagellar agglutinin, although the same temperature for 24 hours inactivated the somatic agglutinin and the cow serum precipitin. Hemolysin deteriorates slowly at 60°C., so that after 4 days the serum, which originally reacted at a dilution of 1:10,240, only titered 1:160. The red cell agglutinin was about as resistant as the hemolysin in that a little still remained at the end of the test period. The experiments while incomplete add further proof that the somatic agglutinin and the precipitin are the least resistant to heat, while the flagellar agglutinin is on the whole comparatively stable. A final experiment was performed to determine, if possible, at what temperature rabbit serum globulin was inactivated. With this in view, globulin was obtained by precipitation with ammonium sulfate, and a series of guinea pigs and chickens received several intraperitoneal injections. In no instance was a globulin precipitin obtained. By immunizing fowls in a similar manner with small quantities of rabbit serum good precipitin was obtained. The diluted rabbit sera heated at various temperatures for 20 minutes were tested for their antigenic activity with rabbit serum precipitin. It was found that diluted serum heated to 90°C. for 20 minutes reacted to about the same antigenic level as that not heated. Even boiling for 20 minutes failed to reduce greatly its antigenic properties. Paradoxically the visibility of the reaction was more intense with the antigen heated at the higher temperatures. The phenomenon was altogether so opposed to the usual conceptions of the inactivation of antigens that the subject will be gone into with more detail in a later communication. Although it is not possible to show definitely in the experiments that the globulin is or is not inactivated at certain temperatures, nevertheless it appears probable that certain of the antibodies are destroyed at temperatures below that capable of greatly altering globulin. It must be recognized that comparisons cannot be made between similar antibodies in the serum of different species, since somatic agglutinin in rabbit serum resisted 70°C. for 20 minutes, although the same agglutinin in cow serum was destroyed at 65°C.     

THE EFFECT OF FORMALDEHYDE ON PNEUMOCOCCI

When used in low concentration, formaldehyde increases the rate of autolytic disintegration of pneumococci whereas in large concentrations it completely inhibits autolysis and preserves both the morphological and staining characteristics of the cells. Pneumococci treated with large concentrations of formaldehyde, then washed free of the antiseptic and resuspended in physiological solutions, rapidly undergo a change which renders them Gram-negative and smaller. The lysis is only partial, however, and is not accompanied by an actual disintegration of the cell. It is caused by the autolytic enzyme of the cell which remains inactive in the presence of an excess of formaldehyde but recovers its activity when the cells are resuspended in a neutral medium after removal of the antiseptic. If the autolytic enzyme is irreversibly inactivated by heating, or maintained inactive in acid or alkaline reaction, the formolized cells retain their staining characteristics and morphological integrity. Formolized pneumococci which have become Gram-negative owing to the action of their autolytic enzyme, fail to elicit the type specific carbohydrate antibodies in rabbits. Formolized pneumococci in which the autolytic enzyme has been destroyed or maintained inactive, and which have retained their Gram-positive character, function as a very effective type specific antigen in the rabbit. These observations emphasize once more the close relation between the Gram-positive structure of pneumococci and the capsular polysaccharide antigen of the cell. They can be used as a basis for the preparation of suspensions of formolized pneumococci which are stable and very effective as type specific antigens.