Edwin W. Schultz

Regeneration of Olfactory Cells.

It is generally held that nerve cells which have been destroyed are not restored. While this may well be true for the type of nerve cells which are usually considered in this connection, it has not been demonstrated to apply to the neurons of the olfactory nerve, the cell bodies of which retain the primitive location in the surface epithelium found in certain lower forms of animal life (neuroepithelium). The cell bodies of the olfactory nerve (olfactory cells) are located for the most part in the mid-zone of the fairly thick pseudostratified columnar epithelium of the olfactory mucosa. 1 Each cell gives off two processes: (1) a superficial stout one, or dendrite, which extends slightly beyond the surface of the mucosa and (2) a deep one, or axone, which passes centralward to the olfactory bulbs under the frontal lobe of the brain. These processes can be demonstrated only by special staining procedures, such as the Bodian method for showing neurofibrils, 2 and are less easily stained than the fibrils of other nerves. In previous communications 3 we have reported that it is possible to induce a high degree of resistance in monkeys against intranasal inoculation with the virus of poliomyelitis by a previous irrigation of the nasal passages with a 1% solution of zinc sulfate. The resistance so induced generally lasts at least one month, but is almost without exception followed by a return of susceptibility within the next 3 or 4 months. It is initiated by an extensive coagulation necrosis of the olfactory mucosa, followed usually within a day or two by its separation, more or less en masse, from the underlying lamina propria and adjacent glands of Bowman. 1

Bacteriological Observations on a Case of Meningo-Encephalitis.∗

On December 12, 1933, a nurse became suddenly ill and repeatedly vomited bile stained material. The following morning she had a severe chill, a temperature of 101 °F., severe headache, some rigidity of the neck and a doubtful Kernigs sign. A day later her temperature reached 105.3°F. and the meningeal symptoms were more marked. During the first 10 days of her illness the more prominent symptoms were: fever, headache, occipital tenderness, rigidity of the neck, strabismus, ptosis of left eyelid, diplopia, twitching muscle fasciculi and lethargy from which she could be fairly easily roused. There was gradual improvement after about the tenth day. Ten weeks after the onset there remained some speech defect, difficulty in feeding herself, dysmetria, sprawling gait, slight nodding of head, emotional instability and impaired memory. Spinal fluid obtained December 15 was hazy and contained 812 cells per cu. mm. (95% small lymphocytes). The sugar content was 37 mg. per 100 cc. and the gold curve 0023321111. Subsequent specimens gave a paretic curve. Repeated Wassermann tests have been negative. Inoculated guinea pigs failed to develop tuberculosis. Of special interest is the fact that an apparently new Gram-positive bacillus was obtained in pure culture from the spinal fluid on December 18, 19, 21, 22, 26, 28, 1933; January 3, 8, 10, 18, March 8, April 12, 1934. Cultures made May 10 were negative. A rabbit inoculated intracerebrally with 0.5 cc. of the spinal fluid obtained on December 28 developed a severe encephalitis 2 days after inoculation. The same organism was cultivated from the brain tissue of this animal.

Recent advances in the study of poliomyelitis

Summary 1. The evidence at present indicates that the virus of poliomyelitis is a true ultramicroscopic virus, probably less than 50 μμ. in size. 2. The virus apparently does not yield to cultivation on lifeless culture media, but like other ultramicroscopic viruses is regenerated only in the presence of living tissue cells. 3. As an antigen the virus calls forth antibodies resembling antitoxins more closely than antibacterial antibodies. This suggests that virus may be an aggregate much simpler in composition than a bacterial cell. 4. The pathogenicity of the virus is restricted to man and certain species of monkeys. 5. The virus is highly selective in directing its attack, not only restricting its attack to certain hosts but to certain special cells within these hosts. After reaching the olfactory bulb from the nasopharyngeal vault, it seems to travel exclusively by nerve axons to different levels of the central nervous system, apparently remaining in all stages of the disease in very intimate association with nerve cells. 6. The immunity following recovery from poliomyelitis is of a solid and durable character. This may be determined in part by a persistence of some virus in the tissues of the recovered individual. 7. There is evidence that a large percentage of the adult population has acquired an immunity to poliomyelitis by natural means, probably as the result of a subclinical infection. 8. The value of serum therapy continues to remain an unsettled question, the general impression being that it is of some value if used early enough in the disease. 9. In addition to human convalescent serum for therapeutic purposes, there has become available the serum of certain normal human adults not known to have had the disease, and also the serum of individual poliomyelitis refractory animals which have been subjected to virus injections over a period of time. The serum of individual sheep, goats, and especially horses may attain an antibody value comparable to that of the average poliomyelitis convalescent serum. 10. Actual solution of the poliomyelitis problem seems to rest on a systematic active immunization of children and young adults by appropriate methods still to be worked out. Such preventive measures also call for some convenient, inexpensive, and reliable means of detecting those who require artificial immunization.

Further observations on the cultivation of strains of poliomyelitis virus in developing eggs.

Summary Further observations on the cultivation of the murine SK strain of poliomyelitis virus in fertile eggs are reported. This strain has now been carried through 30 passages in eggs. The virus has been found to be widely distributed in the infected embryo. It is easily transmitted from egg to egg by different routes. Embryos from 5 to about 14 days of age seem to serve equally well for the propagation of the virus, but those above 16 days of age fail to support its growth. The murine MM strain was carried through 10 egg passages; a Stanford mouse passage strain, believed to be a high titer variant of the Lansing strain, tentatively labeled C(M) virus, was carried through 15 egg passages. The GD VII strain of Theilers mouse encephalomyelitis virus was carried through 10 passages. It caused the lowest incidence of embryo deaths of the three viruses cultivated. Five strains of poliomyelitis virus failed to show growth in eggs.

Cultivation of the Murine SK Strain of Poliomyelitis Virus in Developing Eggs.

Summary The murine SK strain of poliomyelitis virus was carried through 14 passages on developing eggs without evident diminution of virus content. Its multiplication was associated with a high incidence of deaths in the embryos. The incidence of deaths was influenced by the temperature at which the eggs were incubated after inoculation.†

Electron microscopic observations on Pseudomonas aeruginosa bacteriophage.

Summary A sperm-shaped structure was identified with two bacteriophages active for Ps. aeruginosa.

A gelatinous variant of Pseudomonas aeruginosa.

Summary What appears to be a new variant of Pseudomonas aeruginosa is described. It is characterized by the production of large amounts of gelatinous material when grown on media containing glycerol, and on solid media gives rise to markedly raised colonies of a stiff jelly-like consistency and to a thick gelatinous layer near the surface of liquid media. The variation seems to be entirely one of ability to utilize glycerol for the production of the particularly gelatinous material, and to be unrelated to any one of the several basic colony forms known to occur in this species.

Listerella Monocytogenes: A Cause of Meningo-Encephalitis in Man.

In a preliminary note 1 we reported the isolation from a human case of meningo-encephalitis of a new organism which we identified with the genus Corynebacterium. We wish at this tme to report additional observations and to correct the classification of the organism. The clinical history of the case is described in the preliminary note and has been more fully presented by Marcellus, Crouch and Terry. 2 A noteworthy fact is that the organism was cultured from the spinal fluid of this patient 12 times over a period of nearly 4 months, though the period of acute illness lasted less than 2 weeks. A month after our report, Burn 3 in Connecticut reported the isolation of a similar organism from 3 fatal cases of meningo-encephalitis in infants. Later, 4 he reported a fourth fatal case, this time in an adult, and identified his strains with the genus Listerella. In this paper he refers also to a case reported to him by Dr. W. Allen of Hartford, Connecticut. From a fatal case of meningitis in a human adult in Scotland, Gibson 5 isolated an organism which he thought belonged to the genus Corynebacterium. Morphologically and culturally the organism resembled our strain. In Norway, Tesdal 6 described a case of fatal meningitis in man “caused by a Corynebacterium.” His description of the organism, however, suggests a Listerella. More recently Carey 7 in Massachusetts has isolated a Listerella from a case of acute cerebrospinal meningitis in a boy. This patient recovered. These seem to comprise all the cases of human infection known to have been caused by Listerella. Since publishing our preliminary note 1 our organism has been definitely identified as Listerella monocytogenes, originally described by Murray, Webb and Swann 8 and isolated by them from a spontaneous disease in rabbits characterized by monocytosis. Similar organisms have also been recovered from a variety of diseases in animals, including a plague-like disease in gerbille in South Africa, meningo-encephalitis in sheep and in cattle, and a disease in chickens associated with massive myocardial necrosis (for a review see Seastone 9 and Jungherr 10 ).

Infectivity of Murine SK Strain of Poliomyelitis Virus.

Summary Observations are reported on the infectivity of the murine SK strain for mice based on certain types of exposure to the virus. Some of the infected mice were embedded in toto, sectioned, and examined histologically for evidence of virus multiplication in non-nervous as well as nervous tissue. Although infections occurred readily under various types of exposures to the virus, no lesions suggestive of virus activity were observed in any tissues or organs apart from the CNS. Infections by the intranasal route could not be prevented by prior intranasal instillations of zinc sulphate solution used to block the olfactory pathway as a portal of entry. Mice could be readily infected with virus-contaminated drinking water. Virus cultivated in developing eggs proved less infectious for mice when supplied to the animals in drinking water or food than mouse brain-cord virus suspensions. Old mice showed themselves less susceptible than young mice to certain types of exposure, but to other types of exposure the difference was less marked. Young mice could be readily infected by housing them in battery jars contaminated with virus, but it required a heavy contamination to bring this about. While a virus suspension dropped into the conjunctiva after light scarification of the cornea induced infection in 70% of mice, only 10% infection resulted when this procedure was not preceded by scarification of the cornea. Infection was induced in 60% of mice by applying the virus to the skin and then introducing it by the multiple puncture method. Infection was induced in 30% of mice on introducing the virus into the rectum. Young mice can be easily infected by the intraperitoneal route, this route being only slightly less sensitive than the intracranial route. The average incubation period following intravenous inoculation did not differ significantly from that observed following inoculations by other routes with similar amounts of virus. The virus induced a typical poliomyelitic syndrome in guinea pigs. In these animals it became a low titer virus. It proved infectious for hamsters but not for rabbits or monkeys. Attempts to adapt it to baby chicks were unsuccessful.

Antipoliomyelitis Serum Production in the Horse.

In a previous note 1 we reported our inability to demonstrate an antibody response in poliomyelitis refractory animals in any way comparable to that realized in monkeys. Although antisera produced in the guinea pig, rabbit, dog, sheep and goat in some instances inactivated an equal volume of a 5% suspension of virus tissue (cord and medulla) when these sera were employed in an undiluted form, previous dilution of the serum to 1-2 (after addition of virus suspension, 1-4) failed to render such a virus suspension non-infectious for monkeys. Essentially the same results were obtained with the serum of a horse, which at the time of the previous report (June, 1930) had been given 9 intravenous injections of 50 cc. (one injection 20 cc.) each of a 10% suspension of virus material (cord and medulla) over a period of 45 days. Since the horse represented a much later addition to our series of poliomyelitis refractory animals, virus injections have been continued to make the period of “immunization” more comparable to those of the sheep and goat, which received virus injections over a period of about a year. The results with the horse have, however, proven far more gratifying than in the case of the other poliomyelitis refractory animals. Titrations carried out recently on serum procured from the horse 8 months after the “immunization” was begun indicate that the serum is now capable of inactivating an equal volume of a 5% virus suspension in a final serum dilution of 1-60, which according to the recent observation of Shaughnessy, Harmon and Gordon 2 may equal, if not exceed the antibody titer of the average human poliomyelitis convalescent serum. The antibody titer is not as high as that of some of the hyperimmunized poliomyelitis convalescent monkey sera we have titrated (1-128), but we anticipate that with further immunization of the horse the titer may rise to a higher level.