bluuebunny/crossref_metadata_2025 huggingface dataset:
SECOND ATTACKS OF POLIOMYELITIS
Authors: Howe Howard A. , Bodian David
Published: June 2004

1. It was found in forty rhesus monkeys that intracerebral, intraocular, intracutaneous, intraperitoneal, intraspinal, and neural inoculations of poliomyelitis virus produced no lesions in the olfactory bulbs despite the fact that the animals contracted pronounced paralyses. This indicated that the virus could be restricted to certain neuronal systems.

2. Similarly intranasal inoculation of seven animals produced no lesions in the ciliary ganglia.

3. Two monkeys convalescent from an intracutaneous and an intracerebral inoculation respectively had further paralyses after intranasal inoculation of heterologous virus. A third animal convalescent from an intranasal inoculation showed extension of lesions after intranasal and intraocular inoculation with heterologous virus.

4. Two spinal animals in which an attack of poliomyelitis was limited to an isolated segment of spinal cord, contracted typical paralyses in the previously uninvaded portions of the CNS following intranasal inoculation of homologous virus.

5. Four of six convalescent monkeys showed extension of lesions but no clinical signs after homologous virus inoculation through a previously un-invaded portal.

6. Four animals convalescent from a unilateral intranasal inoculation showed evidence of new invasion in the opposite olfactory bulb but no extension of paralyses following a second inoculation of homologous virus into the appropriate nostril.

7. Two animals had second attacks after heterologous second inoculations. The intranasal portal was employed for both exposures.

8. It thus seems apparent that in the rhesus monkey a second attack of poliomyelitis, whether paralytic or not, seems to depend upon the strains of virus used and the degree to which virus is disseminated through the neuraxis during the first exposure.

9. The above experimental data emphasize the difficulty of utilizing the rhesus monkey for experiments seeking to elucidate the mechanisms of immunity in man and suggest that human immunity to poliomyelitis does not result from immunization of the nervous system but rather is the result of some process which prevents infective quantities of active virus from reaching nervous tissue.
MYOHEMOGLOBINURIA
Authors: Yuile Charles L. , Clark William F.
Published: June 2004

When myohemoglobin is injected intravenously into dogs, in amounts ranging from 0.75 to 1.50 gm., it is rapidly eliminated from the plasma and approximately 65 per cent is excreted by the kidneys in from 1½ to 2½ hours.

Myohemoglobin does not appear in the urine below a threshold plasma concentration which is slightly under 20 mg. per 100 cc. but above this level the rate of renal excretion is directly proportional to the plasma concentration.

The maximum myohemoglobin/creatinine clearance ratio averages 0.58 contrasted with a value of 0.023 for blood hemoglobin. This indicates that the rate of renal clearance of myohemoglobin is twenty-five times more rapid than that of blood hemoglobin. Evidence is presented that the excretory mechanism is essentially similar for the two substances but that differences in molecular weight account for different rates of glomerular filtration.
OBSERVATIONS ON THE INFECTION OF CHICK EMBRYOS WITH BACTERIUM TULARENSE, BRUCELLA, AND PASTEURELLA PESTIS
Authors: Buddingh G. John , Womack Frank C.
Published: June 2004

1. Comparison of the infections of chick embryos by the chorio-allantoic route indicates that Bacterium tularense and Brucella suis, abortus, and melitensis exhibit varying degrees of facultative intracellular parasitism. Pasteurella pestis is adapted to rapid proliferation and spread in the intercellular fluids.

2. In the early stages of infection Bacterium tularense has a marked affinity for growth within ectodermal epithelial cells. Brucella suis and Brucella abortus differ in their selectivity for cells of mesodermal derivation and especially in their effect on vascular endothelium. The strain of Brucella melitensis studied is limited in its intracellular growth to ectodermal epithelium.

3. Many of the features characteristic of these infections in the natural hosts are reproduced in the chick embryo and its membranes.

4. The possible implications regarding the differences in behavior of these microorganisms in relation to the problem of infection and pathogenesis of these diseases are discussed.
STUDIES ON CONDITIONS AFFECTING THE SURVIVAL IN VITRO OF A MALARIAL PARASITE (PLASMODIUM LOPHURAE)
Authors: Trager William
Published: June 2004

The survival of Plasmodium lophurae in vitro, at temperatures of 39.5–42°C., is favored by a balanced salt solution having a high potassium content, by aeration but not by a very high oxygen tension, by an optimal density of parasites per cubic millimeter, by frequent renewal of the suspending medium, by concentrated red cell extract, by optimal concentrations of plasma or serum, of chick embryo extract, of glucose or glycogen, and of glutathione, and probably by yeast extract and a very low concentration of liver extract.

In the best preparations, as judged by infectivity, more than 40 per cent of the parasites were alive on the 3rd day, more than 20 per cent on the 4th day, perhaps 1 per cent on the 5th day, and about 0.05 per cent on the 6th day. Evidence was obtained that the parasites had multiplied during the 1st day of incubation.
COMPARATIVE STUDY OF MENINGOPNEUMONITIS VIRUS, PSITTACOSIS OF PIGEON ORIGIN, AND PSITTACOSIS OF PARROT ORIGIN
Authors: Pinkerton Henry , Moragues Vicente
Published: June 2004

Comparative morphological, histological, and biological studies suggest a close relationship between the meningopneumonitis virus of Francis and Magill and a virus recovered from thiamin-deficient pigeons. Both of these viruses are morphologically identical with typical psittacosis, and it seems probable that they are biologically modified strains of psittacosis. They both differ from typical psittacosis in that they are regularly more pathogenic for the pigeon after intracranial injection, and fail to produce hepatic necrosis after intraperitoneal injection in mice. A virus recently isolated from human cases of atypical pneumonia by Eaton, Beck, and Pearson may also be closely related to these two viruses.

A number of psittacosis viruses of pigeon origin showed a similarly increased pathogenicity for pigeons by the intracerebral route, as compared with psittacosis viruses of parrot origin. The viruses of parrot origin, however, commonly produced latent infection in pigeons even when clinical illness was not evidenced.

For the isolation of psittacosis of pigeon origin from human sputum the intracranial injection of mice or pigeons may be essential, although it is probable that the intranasal injection of mice would be successful. The intraperitoneal injection of mice may give negative results.
STUDIES IN THE PATHOGENESIS OF EXPERIMENTAL DYSENTERY INTOXICATION
Authors: Penner Abraham , Bernheim Alice Ida
Published: June 2004

The intravenous injection of Shiga toxin into dogs causes a rise in hemoglobin, red blood cell count, hematocrit reading, and specific gravity of the whole blood. There is thus a decrease in circulating blood volume. The specific gravity of the blood plasma does not change. These findings indicate that the toxin of B. shigae produces a shock-like circulatory state. As a result there occurs a compensatory vasoconstriction in the duodenum of the dog and in the cecum of the rabbit. It has been shown that the toxin of B. shigae has no direct effect upon the intestinal mucosa when brought into contact therewith, but that its absorption through the mucosa leads to the appearance of a lesion in the duodenum of the dog. Therefore we interpret the pathological alterations in the intestinal tract, following the injection of Shiga toxin, as the anatornic end result of a pronounced and prolonged homeostatic vasoconstriction.
THE FORMATION OF ANTIBODIES IN THE POPLITEAL LYMPH NODE IN RABBITS
Authors: Ehrich W. E. , Harris T. N.
Published: June 2004

Typhoid vaccine and sheep erythrocytes were injected subcutaneously into the feet of rabbits, and the subsequent formation of agglutinins and hemolysins in the popliteal lymph node was compared with the output of lymphocytes through the efferent lymph and with changes in the lymph node.

Antibodies began to appear in the efferent lymph 2 to 4 days after the injection of the antigen and reached their highest titer after 6 days. This was preceded by a sharp rise in the output of lymphocytes through the efferent lymph, while in the lymph node there was lymphatic hyperplasia after preliminary infiltration of granulocytes and monocytes. This hyperplasia was first of a diffuse type, but was later superseded by large so called germinal centers, the latter lagging somewhat behind the rise in antibody titer.

The fact that the tissue response accompanying the formation of antibodies was chiefly a lymphocytic one points to the lymphocyte as a factor in the formation of antibodies.
ORAL PAPILLOMATOSIS OF RABBITS: A VIRUS DISEASE
Authors: Parsons Robert J. , Kidd John G.
Published: June 2004

Papillomas occur frequently on the oral mucosa of domestic rabbits procured in the metropolitan area of New York. They are small and benign, and are situated mostly on the under side of the tongue. A filtrable virus can be extracted from them with which growths can be reproduced in the oral mucosa of several species of rabbits and hares but which fails to cause lesions when inoculated into other rabbit tissues and into the oral mucosa of other species. The virus differs notably from the Shope virus, which causes cutaneous papillomas in rabbits but proves innocuous to oral mucosa: rabbits solidly immune to the oral papilloma virus are fully susceptible to the Shope virus and vice versa.

The oral papillomas are not highly contagious, for susceptible animals kept in individual cages in the same rooms with others carrying the growths, fed the same kind of food, and cared for by the same attendants, do not "catch" them. They are found much more frequently in the offspring of dams that carry the growths than in those of mothers free from them, and the causative virus can be recovered from the mouth washings of rabbits having no growths. The observations indicate that the virus may be spread by transfer from the mother to the young during the period of suckling, and that it may lie latent in the mouth, doing no harm unless the mucous membrane is injured. The slight trauma occurring now and then when coarse foods are chewed may furnish the required tissue nidus under natural conditions, for papillomas occasionally appear after virus has been dropped into the mouths of uninoculated rabbits; but the more extensive injury and healing resulting from experimental tattoo inoculations proves regularly effective in this respect. Tar can also act as an efficient adjuvant to the virus, the incidence of "spontaneous" oral papillomas being much higher in domestic rabbits that had had the opportunity to lick tar from their ears and paws during long periods than in normal control animals. The virus is recoverable in quantity from the oral papillomas of tarred domestic rabbits, and the findings indicate that it is their essential cause, the tar acting merely to prepare the tissue for the virus' action. For the same tar does not elicit oral papillomas in wild cottontail rabbits, which do not carry the causative virus though fully susceptible to it.

The implications of the findings are discussed.
THE COURSE OF EXPERIMENTAL INFECTION OF THE CHICK EMBRYO WITH THE VIRUS OF EQUINE ENCEPHALOMYELITIS
Authors: Bang Frederik B.
Published: June 2004

The titration curve for the virus of Eastern equine encephalomyelitis inoculated into the 10 day old chick embryo shows that the maximum increase in virus content continues until shortly before the generalized destruction of the embryo is apparent. This is followed by a stationary phase.

Histological studies of infected embryos fail to demonstrate selective tissue destruction, and titrations show the virus to be distributed throughout the egg, although concentrated in the embryo.

The chorioallantoic membrane gradually becomes increasingly resistant with age to both the Eastern and Western viruses. Increased resistance with age is also apparent in the hatched chick.

These findings are based on the use of the chick embryo itself as the test animal to determine the 50 per cent mortality end-point. The limits of accuracy of this method are defined.
CHANGES IN LIPID CONTENT OF SERUM AND OF LIVER FOLLOWING BILATERAL RENAL ABLATION OR URETERAL LIGATION
Authors: Winkler A. W. , Durlacher S. H. , Hoff H. E. , Man E. B.
Published: June 2004

1. In the dog and monkey bilateral nephrectomy or ureteral ligation results in a marked, progressive increase of total fatty acids, of free and esterified cholesterol, of phospholipid, and of free fat of serum.

2. No such changes follow unilateral nephrectomy, splenectomy, or fasting.

3. The increase after bilateral nephrectomy is not inhibited by glucose administration.

4. A marked increase of the phospholipid and a less significant elevation in cholesterol content of the liver accompanies this increase of serum lipids after bilateral nephrectomy.
NEUTRALIZING AND COMPLEMENT-FIXING ANTIBODY PRODUCTION AND RESISTANCE FOLLOWING VACCINATION IN EXPERIMENTAL ENCEPHALITIS INFECTIONS
Authors: Casals J.
Published: June 2004

In mice vaccinated subcutaneously with different doses of virulent W.E.E. virus or with formolized vaccine, neutralizing and complement-fixing antibodies paralleled resistance to some extent yet appeared in groups in which resistance remained undetectable, persisted at a similar maximum level in spite of different titers of resistance, and after resistance had become negligible.

In mice vaccinated subcutaneously with different doses of virulent St. Louis encephalitis virus or with formolized vaccine, neutralizing and complement-fixing antibodies bore little relation to resistance. Neutralizing antibodies appeared only in the group showing resistance but not until resistance was diminishing. Complement-fixing antibodies developed equally well in groups with or without resistance.
BIOCHEMICAL STUDIES ON SHOCK
Authors: Russell Jane A. , Long C. N. H. , Wilhelmi Alfred E.
Published: June 2004

1. With increasing severity of shock following hemorrhage in fasted rats there is an increasing depression in the rate of oxygen uptake, in oxygen, of liver slices from the bled animals. The respiration of kidney tissue is only slightly depressed even in severe states of shock.

2. The rates of oxygen uptake of liver tissue from bled rats are nicely correlated with the increases in blood amino nitrogen that follow severe hemorrhage.

3. A supplement of coenzyme factors, in the form of a hot water extract of normal rat liver, increases the oxygen uptake of liver tissue from rats in mild shock, but is without effect on the respiration of liver slices from rats in moderate or severe shock.

4. The ability of rat liver to oxidize succinate is not impaired even in severe shock, but the extra oxygen uptake does not improve the basal rate of respiration of the tissue.

5. Effects on the rate of oxygen uptake of normal rat liver slices comparable to those seen after hemorrhage could be produced by exposing the tissue to an atmosphere of nitrogen for periods of 15 and 60 minutes. This treatment had more marked effects on the respiration of kidney slices than are found after hemorrhage, but the kidney, unlike the liver, exhibited a marked degree of recovery in the presence of glucose.

6. The significance of these findings is briefly discussed.
BIOCHEMICAL STUDIES ON SHOCK
Authors: Engel Frank L. , Harrison Helen C. , Long C. N. H.
Published: June 2004

1. In a series of rats subjected to hemorrhage and shock a high negative correlation was found between the portal and peripheral venous oxygen saturations and the arterial blood pressure on the one hand, and the blood amino nitrogen levels on the other, and a high positive correlation between the portal and the peripheral oxygen saturations and between each of these and the blood pressure.

2. In five cats subjected to hemorrhage and shock the rise in plasma amino nitrogen and the fall in peripheral and portal venous oxygen saturations were confirmed. Further it was shown that the hepatic vein oxygen saturation falls early in shock while the arterial oxygen saturation showed no alteration except terminally, when it may fall also.

3. Ligation of the hepatic artery in rats did not affect the liver's ability to deaminate amino acids. Hemorrhage in a series of hepatic artery ligated rats did not produce any greater rise in the blood amino nitrogen than a similar hemorrhage in normal rats. The hepatic artery probably cannot compensate to any degree for the decrease in portal blood flow in shock.

4. An operation was devised whereby the viscera and portal circulation of the rat were eliminated and the liver maintained only on its arterial circulation. The ability of such a liver to metabolize amino acids was found to be less than either the normal or the hepatic artery ligated liver and to have very little reserve.

5. On complete occlusion of the circulation to the rat liver this organ was found to resist anoxia up to 45 minutes. With further anoxia irreversible damage to this organ's ability to handle amino acids occurred.

6. It is concluded that the blood amino nitrogen rise during shock results from an increased breakdown of protein in the peripheral tissues, the products of which accumulate either because they do not circulate through the liver at a sufficiently rapid rate or because with continued anoxia intrinsic damage may occur to the hepatic parenchyma so that it cannot dispose of amino acids.
THE DURATION OF IMMUNITY TO PLASMODIUM KNOWLESI MALARIA IN RHESUS MONKEYS
Authors: Maier J. , Coggeshall L. T.
Published: June 2004

Rhesus monkeys with experimental Plasmodium knowlesi infections of varying duration were treated with sodium sulfathiazole to sterilize the infection and after differing lengths of time were reinoculated intraperitoneally with homologous strains of the plasmodium, for the purpose of determining whether there is any acquired immunity to malaria in hosts from whom all parasites have been removed.

Two monkeys, one receiving sulfathiazole on the 2nd day of acute infection and the other on the 4th day, had no immunity at the time of reinoculation, 3 weeks and 10 weeks later, respectively. Both developed infections which followed the course usual in an acute attack in a normal monkey.

In monkeys which survived acute infection with the aid of immune serum or quinine and in which a naturally acquired immunity had developed to the point where the acute infection was converted into a chronic one, there was an undoubted persistence of partial immunity up to about a year after sterilization of the infection with sulfathiazole, as indicated by recovery of reinoculated animals after mild or moderately severe infections differing widely in characteristics from the infection in the normal monkey.

The end point at which immunity disappears seems to be independent of the length of the chronic infection.
POLIOMYELITIS IN THE CYNOMOLGUS MONKEY
Authors: Faber Harold K. , Silverberg Rosalie J. , Dong Luther
Published: June 2004

1. Poliomyelitis virus suspensions were atomized so as to produce dry droplet nuclei which, suspended in air, were introduced into a special infecting chamber and inhaled by test animals, both rhesus and cynomolgus monkeys.

2. Without olfactory blockade, 5 of 7 rhesus and 6 of 7 cynomolgus monkeys developed poliomyelitis of the CNS with entry through the olfactory nerves.

3. With olfactory blockade, 2 of 35 rhesus and 4 of 10 cynomolgus monkeys developed this form of the disease by routes proved by serial sections of the olfactory bulbs not to have been olfactory.

4. The neural pathways of infection from the mucous surfaces to the CNS in the 4 cynomolgus monkeys with blockade were shown in 2 instances to have been the afferent fibers of the trigeminal nerve into the Gasserian ganglion and thence to its central connections in the pons-medulla; in another case this was the probable route. In one instance the pathway consisted of the sympathetic fibers of the nose or nasopharynx into the cervical sympathetic ganglia and thence into the uppermost levels of the thoracic cord. The routes in the 2 rhesus monkeys with non-olfactory takes were not accurately determined but in one there was suggestive evidence of entry through the trigeminal nerve.

5. Study of the peripheral ganglia in a number of exposed cynomolgus and rhesus monkeys, including several with no demonstrated involvement of the CNS, revealed lesions most constantly in the Gasserian ganglia; less so in the cervical sympathetics and still less so in the celiac. In 2 rhesus monkeys dying of other causes a few days after exposure, lesions were limited to the Gasserian ganglia. No evidence was found in any case of passage of infection from the celiac ganglia into the CNS.

6. The importance of the peripheral ganglia as intermediate stations in the centripetal passage of infection from the body surface is again emphasized.

7. Comparison of the present with a previous study suggests that infection by inhalation of virus occurs with greater ease than by ingestion.
EXPERIMENTAL TYPHUS INFECTION IN THE EASTERN COTTON RAT (SIGMODON HISPIDUS HISPIDUS)
Authors: Anderson Charles R.
Published: June 2004

1. The course of typhus infection in cotton rats has been described, and a relationship between the dose administered and the time of death has been demonstrated.

2. Rather incomplete pathological studies indicated that typhus in the cotton rat is an acute infection primarily involving mesothelial tissue, and tissues of mesothelial origin. Depending somewhat upon the route of infection, rickettsiae were most easily demonstrated in smears from liver, brain, and pericardial and peritoneal exudates. By subinoculation into the yolk sacs of fertile eggs, rickettsiae were readily isolated from these sources and, with rather less facility, from blood.

3. Although relatively large numbers of rickettsiae were found necessary to produce lethal infections, the susceptibility of the cotton rat as measured by the development of an immunizing infection proved to be very high. The supporting data suggest that cotton rats might be suitable for the isolation of new strains.

4. Contrary to the earlier report (1), age was not found to influence markedly the susceptibility of the cotton rat to lethal infection with any of the strains studied.

5. Comparative studies indicated that cotton rats were just as susceptible to murine infection as white mice and more susceptible to Breinl infection than the latter animal. The susceptibility of cotton rats and of guinea pigs to Breinl infection was found to be about the same. Comparative studies in these animals were not made with other strains.

6. No toxic activity of rickettsial suspensions could be demonstrated for cotton rats, although suspensions of infected cotton rat livers were shown to be toxic for mice.

7. A method has been described by which three strains of Rickettsia prowazeki were carried in serial intracardial passage as fatal infections for cotton rats. No evidence was obtained that prolongation of such passage increased the virulence of the strains for the rats.

8. Cotton rats surviving typhus infection were shown to be solidly immune to reinfection with homologous virus and to possess nearly complete immunity to rickettsiae of heterologous immunologic character. Sera from recovered rats, furthermore, were shown to possess neutralizing, complement-fixing, and antitoxic antibodies but no agglutinins for proteus OX19.

9. The technique of a serum neutralization test in cotton rats has been described in which the test antigen consists of suspensions of infected cotton rat livers. By this technique neutralizing antibody was demonstrated in human, rabbit, guinea pig, and cotton rat sera. Although some degree of cross-neutralization could be shown, serum titers against homologous antigen were uniformly greater than against comparable doses of antigen of heterologous immune nature.
STUDIES ON THE MAMMARY TUMORS OF DOGS
Authors: Huggins Charles , Moulder Peter Vincent
Published: June 2004

Spontaneous mammary tumors occur in middle aged or senile dogs and are of three principal pathological types; a diffuse epithelial and connective tissue overgrowth often with associated cartilage and bone; solid masses of epithelial cells; intracystic papillomatous tumors. The metastases in this series were always composed of papillary carcinomatous cysts; this tissue in transplants to abdominal connective tissue induced epithelial osteogenesis. All of the dogs with mammary tumors had lipid-rich suprarenal cortical tumors.

The tumors frequently secrete a dilute milk with citric acid content considerably increased above serum levels; large numbers of colostrum corpuscles are present in this fluid. After weaning, normal parturient dogs undergo a complete cessation of mammary secretion; lactation persists considerably longer in dogs with mammary tumor than in normal dogs. Absence of suckling does not eliminate lactation in pseudopregnancy.

The failure of the breast to lactate under the stimulus of prolactin signifies a lack of physiological maturity. The breasts of mature dogs have two types of response to ovariectomy; either great physiological involution results or lactation, actual or potential, is retained in areas for at least 3 to 6 months. Those in which excision of the ovaries does not cause prompt and great involution of the breast include all dogs with mammary tumors, all with pseudopregnancy, and certain dogs, presumably normal which we are unable to characterize further at this time. Excision of the suprarenal glands in addition to the ovaries usually did not completely eliminate lactation or the lactational potential.
THE CARDIOTOXIC ACTION OF PREPARATIONS CONTAINING THE OXYGEN-LABILE HEMOLYSIN OF STREPTOCOCCUS PYOGENES
Authors: Bernheimer Alan W. , Cantoni G. L.
Published: June 2004

1. A study has been made of the effect on the isolated frog's heart of a preparation derived from the supernatant fluid of cultures of hemolytic streptococcus.

2. The streptococcal preparation was found to induce systolic contracture, the contracture usually developing only after the second of two administrations of the preparation. It was found that a single application of the streptococcal preparation sensitized the heart to a second application.

3. The properties of the cardiotoxic factor so far as they have been studied were found to be identical with those of the oxygen-labile hemolysin of streptococci. The capacity of normal and of immune sera to neutralize the cardiotoxic action paralleled the antihemolytic potency of the sera.
TOLERANCE TO AMINO ACID MIXTURES AND CASEIN DIGESTS GIVEN INTRAVENOUSLY
Authors: Madden S. C. , Woods R. R. , Shull F. W. , Remington J. H. , Whipple G. H.
Published: June 2004

Several synthetic mixtures of natural and racemic crystalline amino acids suitable for the daily nitrogen requirement are tested in dogs for their tolerance upon intravenous injection.

Certain mixtures of the ten essential amino acids plus non-essential amino acids exclusive of glutamic acid are accepted without any obvious sign of disturbance even at rates above 10 mg. nitrogen per kilo per minute for quantities greater than 300 mg. per kilo. One such mixture consists in parts per 100 of dl-threonine 7, dl-valine 15, l(-)-leucine 10.9, dl-isoleucine 9.9, l(+)-lysine· HCl·H2O 10.9, dl-tryptophane 3, dl-phenylalanine 9.9, dl-methionine 6, l(+)-histidine·HCl·H2O 5, l(+)-arginine-HCl 5, glycine 9.9, dl-α-alanine 4, dl-serine 2, l(-)-cystine 0.5, and l(-)-tyrosine 1. In addition other well tolerated mixtures included the prolines.

When glutamic acid, natural or racemic, is included in similar mixtures vomiting reactions frequently occur at nitrogen rates above 4 mg. per kilo per minute. Vomiting almost always occurs on the first daily injection containing glutamic acid and usually on any subsequent injection containing more than 100 mg. glutamic acid per kilo unless given very slowly.

Upon the addition of glycine certain mixtures of the ten essential amino acids show an improved tolerance.

Two casein digests tested usually produced vomiting at injection rates above 2 mg. nitrogen per kilo per minute, probably because of their glutamic acid content.

No serious reaction has ever occurrred to any mixture of amino acids or casein digest tested. Elimination of minor reactions such as vomiting appears possible and desirable for greater usefulness of these solutions in parenteral feeding.
A PROTEOLYTIC ENZYME PRODUCED BY GROUP A STREPTOCOCCI WITH SPECIAL REFERENCE TO ITS EFFECT ON THE TYPE-SPECIFIC M ANTIGEN
Authors: Elliott Stuart D.
Published: June 2004

1. Group A streptococci sometimes produce in broth culture an extracellular proteolytic enzyme.

2. Under suitable cultural conditions the enzyme has been demonstrated in representative cultures of most of the Griffith types. Its production by a given strain may be suppressed by serial passage through mice and the variant so produced has been found to maintain this change in character on subculture in artificial media.

3. Under certain conditions, the enzyme attacks the type-specific M antigens of all the group A streptococci so far tested, with the exception of that of type 28. The enzyme exhibits its maximal activity at 37°C.: Extracts made from enzyme-producing cultures which have been grown at this temperature lack the M antigen; enzyme-producing strains may sometimes be induced to yield M substance in extracts by culturing the streptococci at 22° C. Cultures which, when grown at 37° C. yield M substance in extracts, do not produce the enzyme.

4. Human and rabbit fibrin are attacked and streptococcal fibrinolysin is also inactivated by the enzyme. Other susceptible substrates include casein, milk, gelatin, and benzoyl-l-arginineamide but not l-leucylglycylglycine.

5. The general properties of the enzyme resemble those of papain and some of the cathepsins: It is active under the reducing conditions produced in broth cultures by the presence of living bacteria; it is also activated by substances which reduce disulfide to sulfhydryl groups, e.g. potassium cyanide, cysteine, glutathione, and thioglycollic acid, but it is not activated by ascorbic acid. The enzyme is inactivated by iodoacetic acid and also by normal rabbit or mouse serum.
THE EXPERIMENTAL INFECTION OF THE HUMAN BODY LOUSE, PEDICULUS HUMANUS CORPORIS, WITH MURINE AND EPIDEMIC LOUSE-BORNE TYPHUS STRAINS
Authors: Snyder J. C. , Wheeler C. M.
Published: June 2004

Experiments are described which demonstrate that human body lice (Pediculus humanus corporis), were infected experimentally with murine and epidemic louse-borne strains of typhus fever by feeding on suitably prepared rabbits. Details of the two methods of infection, the "bleb technique" and the "I.V. technique," are presented.

It is concluded that the experimental infection of human lice with typhus can be accomplished very easily and rapidly with these methods. The possible applications of the method are discussed.
BACTERIOSTATIC EFFECT OF HUMAN SERA ON GROUP A STREPTOCOCCI
Authors: Rothbard Sidney
Published: June 2004

1. Type-specific antibodies were demonstrated by the indirect bacteriostatic test in sera from human adults convalescing from group A streptococcal infection of the upper respiratory tract. The time of appearance of the antibodies varied from 3 to 5 weeks; and they persisted in 2 patients for at least 37 weeks after the onset of the infection.

2. The specificity of the antibody response in one serum was tested with strains of 7 heterologous types; in another, with 6; and in the third, with 2; but in no instance were cross-reactions observed. Moreover, each convalescent serum showed approximately equal bacteriostasis for 7 different strains of the same type as that which caused the infection.

3. The antibodies were specifically absorbed from the serum by homologous heat-killed streptococci, but not significantly by strains of heterologous types.

4. The specific M antigen of the streptococcal cell with its respective antibody, and not the T substance, appeared to be concerned in the reaction.

5. In spite of numerous technical difficulties inherent in the method, this bacteriostatic test provides a useful procedure for studying type-specific immunity in streptococcal infections.
THE ETIOLOGY OF COLORADO TICK FEVER
Authors: Florio Lloyd , Stewart Mabel O. , Mugrage Edward R.
Published: June 2004

1. The infectious agent of Colorado tick fever filtered through 181 mµ membranes caused infection in hamsters and one volunteer.

2. In five out of seven instances, hamsters were infected initially with serum filtered through 24 mµ membranes or the tick fever appeared on serial transfer.

3. We could not infect two volunteers with 24 mµ filtrates, though these filtrates did cause infection in hamsters as shown by development of the fever on serial transfer. In one instance, the serum of the hamsters rendered ill in this way was injected into the volunteer who had remained well after inoculation with the original material and it caused Colorado tick fever.

4. Normal hamster serum was passaged through 10 groups of animals and then two human volunteers were injected with it. They did not come down with Colorado tick fever, although both of them were susceptible to the disease as shown by the fact that they fell ill of it on later inoculation with serum from a natural instance of the disease.

5. The infectious agent of Colorado tick fever passes through 24 mµ membranes. On this basis, it is classified as a virus.
Unique Inner Pore Properties of BK Channels Revealed by Quaternary Ammonium Block
Authors: Li Weiyan , Aldrich Richard W.
Published: June 2004

Potassium channels have a very wide distribution of single-channel conductance, with BK type Ca2+-activated K+ channels having by far the largest. Even though crystallographic views of K+ channel pores have become available, the structural basis underlying BK channels' large conductance has not been completely understood. In this study we use intracellularly applied quaternary ammonium compounds to probe the pore of BK channels. We show that molecules as large as decyltriethylammonium (C10) and tetrabutylammonium (TBA) have much faster block and unblock rates in BK channels when compared with any other tested K+ channel types. Additionally, our results suggest that at repolarization large QA molecules may be trapped inside blocked BK channels without slowing the overall process of deactivation. Based on these findings we propose that BK channels may differ from other K+ channels in its geometrical design at the inner mouth, with an enlarged cavity and inner pore providing less spatially restricted access to the cytoplasmic solution. These features could potentially contribute to the large conductance of BK channels.
The Bone Morphogenetic Protein Type Ib Receptor Is a Major Mediator of Glial Differentiation and Cell Survival in Adult Hippocampal Progenitor Cell Culture
Authors: Brederlau A. , Faigle R. , Elmi M. , Zarebski A. , Sjöberg S. , Fujii M. , Miyazono K. , Funa K.
Published: June 2004

Bone morphogenetic proteins (BMPs) act as growth regulators and inducers of differentiation. They transduce their signal via three different type I receptors, termed activin receptor-like kinase 2 (Alk2), Alk3, or bone morphogenetic protein receptor Ia (BMPRIa) and Alk6 or BMPRIb. Little is known about functional differences between the three type I receptors. Here, we have investigated consequences of constitutively active (ca) and dominant negative (dn) type I receptor overexpression in adult-derived hippocampal progenitor cells (AHPs). The dn receptors have a nonfunctional intracellular but functional extracellular domain. They thus trap BMPs that are endogenously produced by AHPs. We found that effects obtained by overexpression of dnAlk2 and dnAlk6 were similar, suggesting similar ligand binding patterns for these receptors. Thus, cell survival was decreased, glial fibrillary acidic protein (GFAP) expression was reduced, whereas the number of oligodendrocytes increased. No effect on neuronal differentiation was seen. Whereas the expression of Alk2 and Alk3 mRNA remained unchanged, the Alk6 mRNA was induced after impaired BMP signaling. After dnAlk3 overexpression, cell survival and astroglial differentiation increased in parallel to augmented Alk6 receptor signaling. We conclude that endogenous BMPs mediate cell survival, astroglial differentiation and the suppression of oligodendrocytic cell fate mainly via the Alk6 receptor in AHP culture.
Cbl-dependent Ubiquitination Is Required for Progression of EGF Receptors into Clathrin-coated Pits
Authors: Stang Espen , Blystad Frøydis D. , Kazazic Maja , Bertelsen Vibeke , Brodahl Tonje , Raiborg Camilla , Stenmark Harald , Madshus Inger Helene
Published: June 2004

Ligand binding causes the EGF receptor (EGFR) to become ubiquitinated by Cbl upon association with the adaptor protein Grb2. We have investigated the role of ubiquitin and Grb2 in ligand-induced endocytosis of the EGFR. Incubation of cells with EGF on ice caused translocation of Grb2 and Cbl from the cytosol to the rim of coated pits. Grb2 with point mutations in both SH3 domains inhibited recruitment of the EGFR to clathrin-coated pits, in a Ras-independent manner. On overexpression of the Cbl-binding protein Sprouty, ubiquitination of the EGFR was inhibited, the EGFR was recruited only to the rim of coated pits, and endocytosis of the EGFR was inhibited. Conjugation-defective ubiquitin similarly inhibited recruitment of EGF-EGFR to clathrin-coated pits. Even though this does not prove that cargo must be ubiquitinated, this indicates the importance of interaction of ubiquitinated protein(s) with proteins harboring ubiquitin-interacting domains. We propose that Grb2 mediates transient anchoring of the EGFR to an Eps15-containing molecular complex at the rim of coated pits and that Cbl-induced ubiquitination of the EGFR allows relocation of EGFR from the rim to the center of clathrin-coated pits.
A Novel Intermediate in Initiation Complex Assembly for Fission Yeast DNA Replication
Authors: Yamada Yoshiki , Nakagawa Takuro , Masukata Hisao
Published: June 2004

Assembly of initiation factors on individual replication origins at onset of S phase is crucial for regulation of replication timing and repression of initiation by S-phase checkpoint control. We dissected the process of preinitiation complex formation using a point mutation in fission yeast nda4-108/mcm5 that shows tight genetic interactions with sna41+/cdc45+. The mutation does not affect loading of MCM complex onto origins, but impairs Cdc45-loading, presumably because of a defect in interaction of MCM with Cdc45. In the mcm5 mutant, however, Sld3, which is required for Cdc45-loading, proficiently associates with origins. Origin-association of Sld3 without Cdc45 is also observed in the sna41/cdc45 mutant. These results suggest that Sld3-loading is independent of Cdc45-loading, which is different from those observed in budding yeast. Interestingly, returning the arrested mcm5 cells to the permissive temperature results in immediate loading of Cdc45 to the origin and resumption of DNA replication. These results suggest that the complex containing MCM and Sld3 is an intermediate for initiation of DNA replication in fission yeast.
JAM-C Is a Component of Desmosomes and a Ligand for CD11b/CD18-mediated Neutrophil Transepithelial Migration
Authors: Zen Ke , Babbin Brian A. , Liu Yuan , Whelan John B. , Nusrat Asma , Parkos Charles A.
Published: June 2004

Neutrophil (PMN) transepithelial migration is dependent on the leukocyte β2integrin CD11b/CD18, yet the identity of epithelial counterreceptors remain elusive. Recently, a JAM protein family member termed JAM-C was implicated in leukocyte adhesive interactions; however, its expression in epithelia and role in PMN-epithelial interactions are unknown. Here, we demonstrate that JAM-C is abundantly expressed basolaterally in intestinal epithelia and localizes to desmosomes but not tight junctions. Desmosomal localization of JAM-C was further confirmed by experiments aimed at selective disruption of tight junctions and desmosomes. In assays of PMN transepithelial migration, both JAM-C mAbs and JAM-C/Fc chimeras significantly inhibited the rate of PMN transmigration. Additional experiments revealed specific binding of JAM-C to CD11b/CD18 and provided evidence of other epithelial ligands for CD11b/CD18. These findings represent the first demonstration of direct adhesive interactions between PMN and epithelial intercellular junctions (desmosomes) that regulate PMN transepithelial migration and also suggest that JAM-C may play a role in desmosomal structure/function.
Role of the α-Glucanase Agn1p in Fission-Yeast Cell Separation
Authors: Dekker Nick , Speijer Dave , Grün Christian H. , van den Berg Marlene , de Haan Annett , Hochstenbach Frans
Published: June 2004

Cell division in the fission yeast Schizosaccharomyces pombe yields two equal-sized daughter cells. Medial fission is achieved by deposition of a primary septum flanked by two secondary septa within the dividing cell. During the final step of cell division, cell separation, the primary septum is hydrolyzed by an endo-(1,3)-β-glucanase, Eng1p. We reasoned that the cell wall material surrounding the septum, referred to here as the septum edging, also must be hydrolyzed before full separation of the daughter cells can occur. Because the septum edging contains (1,3)-α-glucan, we investigated the cellular functions of the putative (1,3)-α-glucanases Agn1p and Agn2p. Whereas agn2 deletion results in a defect in endolysis of the ascus wall, deletion of agn1 leads to clumped cells that remained attached to each other by septum-edging material. Purified Agn1p hydrolyzes (1,3)-α-glucan predominantly into pentasaccharides, indicating an endo-catalytic mode of hydrolysis. Furthermore, we show that the transcription factors Sep1p and Ace2p regulate both eng1 and agn1 expression in a cell cycle-dependent manner. We propose that Agn1p acts in concert with Eng1p to achieve efficient cell separation, thereby exposing the secondary septa as the new ends of the daughter cells.
Flagellar Radial Spokes Contain a Ca2+-stimulated Nucleoside Diphosphate Kinase
Authors: Patel-King Ramila S. , Gorbatyuk Oksana , Takebe Sachiko , King Stephen M.
Published: June 2004

The radial spokes are required for Ca2+-initiated intraflagellar signaling, resulting in modulation of inner and outer arm dynein activity. However, the mechanochemical properties of this signaling pathway remain unknown. Here, we describe a novel nucleoside diphosphate kinase (NDK) from the Chlamydomonas flagellum. This protein (termed p61 or RSP23) consists of an N-terminal catalytic NDK domain followed by a repetitive region that includes three IQ motifs and a highly acidic C-terminal segment. We find that p61 is missing in axonemes derived from the mutants pf14 (lacks radial spokes) and pf24 (lacks the spoke head and several stalk components) but not in those from pf17 (lacking only the spoke head). The p61 protein can be extracted from oda1 (lacks outer dynein arms) and pf17 axonemes with 0.5 M KI, and copurifies with radial spokes in sucrose density gradients. Furthermore, p61 contains two classes of calmodulin binding site: IQ1 interacts with calmodulin-Sepharose beads in a Ca2+-independent manner, whereas IQ2 and IQ3 show Ca2+-sensitive associations. Wild-type axonemes exhibit two distinct NDKase activities, at least one of which is stimulated by Ca2+. This Ca2+-responsive enzyme, which accounts for ∼45% of total axonemal NDKase, is missing from pf14 axonemes. We found that purified radial spokes also exhibit NDKase activity. Thus, we conclude that p61 is an integral component of the radial spoke stalk that binds calmodulin and exhibits Ca2+-controlled NDKase activity. These observations suggest that nucleotides other than ATP may play an important role in the signal transduction pathway that underlies the regulatory mechanism defined by the radial spokes.
EST clustering error evaluation and correction
Authors: Wang Ji-Ping Z. , Lindsay Bruce G. , Leebens-Mack James , Cui Liying , Wall Kerr , Miller Webb C. , dePamphilis Claude W.
Published: June 2004

Motivation: The gene expression intensity information conveyed by (EST) Expressed Sequence Tag data can be used to infer important cDNA library properties, such as gene number and expression patterns. However, EST clustering errors, which often lead to greatly inflated estimates of obtained unique genes, have become a major obstacle in the analyses. The EST clustering error structure, the relationship between clustering error and clustering criteria, and possible error correction methods need to be systematically investigated.

Results: We identify and quantify two types of EST clustering error, namely, Type I and II in EST clustering using CAP3 assembling program. A Type I error occurs when ESTs from the same gene do not form a cluster whereas a Type II error occurs when ESTs from distinct genes are falsely clustered together. While the Type II error rate is <1.5% for both 5′ and 3′ EST clustering, the Type I error in the 5′ EST case is ∼10 times higher than the 3′ EST case (30% versus 3%). An over-stringent identity rule, e.g., P ≥ 95%, may even inflate the Type I error in both cases. We demonstrate that ∼80% of the Type I error is due to insufficient overlap among sibling ESTs (ISO error) in 5′ EST clustering. A novel statistical approach is proposed to correct ISO error to provide more accurate estimates of the true gene cluster profile.

Availability: We have automated the methods developed in this paper in a web-based software ESTstat at http://cwdg5.bio.psu.edu/eststat.

Supplementary information:  http://cwdg5.bio.psu.edu/eststat
Identifying Candidate Causal Variants Responsible for Altered Activity of the ABCB1 Multidrug Resistance Gene
Authors: Soranzo Nicole , Cavalleri Gianpiero L. , Weale Michael E. , Wood Nicholas W. , Depondt Chantal , Marguerie Richard , Sisodiya Sanjay M. , Goldstein David B.
Published: June 2004

The difficulty of fine localizing the polymorphisms responsible for genotype-phenotype correlations is emerging as an important constraint in the implementation and interpretation of genetic association studies, and calls for the definition of protocols for the follow-up of associated variants. One recent example is the 3435C>T polymorphism in the multidrug transporter gene ABCB1, associated with protein expression and activity, and with several clinical conditions. Available data suggest that 3435C>T may not directly cause altered transport activity, but may be associated with one or more causal variants in the poorly characterized stretch of linkage disequilibrium (LD) surrounding it. Here we describe a strategy for the follow-up of reported associations, including a Bayesian formalization of the associated interval concept previously described by Goldstein. We focus on the region of high LD around 3435C>T to compile an exhaustive list of variants by (1) using a relatively coarse set of marker typings to assess the pattern of LD, and (2) resequencing derived and ancestral chromosomes at 3435C>T through the associated interval. We identified three intronic sites that are strongly associated with the 3435C>T polymorphism. One of them is associated with multidrug resistance in patients with epilepsy (χ2 = 3.78, P = 0.052), and sits within a stretch of significant evolutionary conservation. We argue that these variants represent additional candidates for influencing multidrug resistance due to P-glycoprotein activity, with the IVS 26+80 T>C being the best candidate among the three intronic sites. Finally, we describe a set of six haplotype tagging single-nucleotide polymorphisms that represent common ABCB1 variation surrounding 3435C>T in Europeans.