Accumulation of acid-fast lipochrome bodies in glial cells of the midbrain nigral lesion in Parkinson's disease.

To confirm or refute the proposed link between nocardiae and Parkinson's disease (PD), we investigated the presence of acid-fast spherical structures similar to filterable nocardiae at the midbrain nigral lesions of three patients with PD. Many clusters of acid-fast lipochrome bodies were dense around blood vessels in the two patients with Hoehn and Yahr stage II and III PD. These clusters were present in the vicinity of melanin-pigmented neurons in the three PD patients studied. Examination of adjacent hematoxylin-and-eosin-stained sections indicated that they consisted of yellow-green granules, bodies, and aggregates in ballooned glial cells. On the other hand, no clusters of acid-fast lipochrome bodies were observed at the compacta region of three control patients. Our results suggest that the immunological and genetic relationship between the acid-fast lipochrome bodies and filterable nocardiae should be investigated.

Tinctorial properties of spherical bodies in broth cultures of Nocardia asteroides GUH-2.

In yeast extract-supplemented brain heart infusion (BHI) broth cultures of Nocardia asteroides GUH-2, many spherical bodies (SBs) were frequently seen nearby filamentous cells. They showed no gram-positivity when Gram stain was applied. When acridine orange stain was applied, many of them showed different green fluorescence from bright orange fluorescence of the filamentous nocardiae under ultraviolet light. Their acid-fastness appeared to depend on the presence of paraffin. Using the polymerase chain reaction (PCR) method, 16S rRNA genes were detected in SB-containing broth cultures inoculated with culture filtrates from broth cultures of the strain and identical to that of N. asteroides. These results suggest that SBs are cell wall-defective (CWD) forms which result from the spontaneous mutation of N. asteroides GUH-2.

Acid-fastness of Nocardia asteroides GUH-2 cultured in brain heart infusion broth supplemented with paraffin.

Nocardia asteroides GUH-2 was not acid-fast when grown in brain heart infusion (BHI) broth. When grown in BHI broth supplemented with paraffin, many filamentous cells showed acid-fastness after treatment with 1% acid-alcohol as the decolorizing agent. When treated with 3% acid-alcohol, filamentous cells were not acid-fast. In addition to the acid-fast filamentous cells of nocardiae, unknown acid-fast spherical bodies were observed in the paraffin-supplemented BHI broth cultures.

Circulating antibody to Nocardia in the serum of patients with Parkinson's disease.

Nocardia is a genus of aerobic Gram-positive bacteria which forms filamentous cells that fragmented into rod-shaped or coccoid elements or L-forms. According to the literature N. asteroides causes encephalitis with parkinsonian features, and it may be an unknown etiologic agent causing encephalitis with a parkinsonian syndrome. Because there are no reliable immunologic tests routinely used for diagnosing nocardial infection, less severe CNS infections with nocardiae may not be recognized or may be attributed to unknown or incorrect etiologic agents. Recently, it was shown that sublethal doses of N. asteroides causes an L-DOPA-responsive movement disorder with Lewy-like bodies in mice. In this study, we detected antibodies to nocardia in the serum of patients with Parkinson's disease (PD). Sera was diluted in PBS and added in two-fold dilutions to coccoid and rod-shaped cells of nocardia attached to nonfluorescence slides. IF test demonstrated antibodies to coccoid and rod-shaped cells of nocardia in the serum from 20/20 patients with PD at a titer greater than 1:10, and 14 controls showed 10 positively. The results suggested that not only PD patients but also age-matched healthy volunteers are routinely exposed to and naturally infected with nocardia-related microorganisms. A further reliable immunologic test will be required.

L-dopa-responsive movement disorder caused by Nocardia asteroides localized in the brains of mice.

Nocardia asteroides can cause infections in the brain of humans and a variety of animals. In mice, invasion of the central nervous system results in specific neurologic signs. Following intravenous injection of various doses of log-phase N. asteroides GUH-2 into female BALB/c mice, localization and growth of nocardial cells within the brains were determined, histopathological sections were prepared, and Nissl substance and tyrosine hydroxylase immunoreactivity were observed. Mice were monitored for the development of neurologic signs, and their responsiveness to L-dopa was determined. It was shown that nocardial cells became localized within specific regions of the brain and then underwent rapid growth followed by a delayed clearance, and there was no inflammatory response at the site of invasion for 24 h. Mice that received a subclinical dose of nocardiae developed specific neurologic signs that emerged following the elimination of nocardial cells from the brain. On the basis of the specific signs, mice could be divided into distinct groups. One group consisted of animals that had a form of hemiparesis that did not respond to L-dopa. They expressed a deviation of the head and a tendency to roll, and when suspended by the tail they would spin rapidly. The second group of mice developed a rhythmic, uncontrolled vertical shake of the head (four to five times per s) tremulous movement, stooped posture, restlessness, and no signs of hemiparesis. The head shakes were temporarily stopped by treatment with L-dopa. Mice that expressed head shakes had a loss of Nissl substance and tyrosine hydroxylase immunoreactivity in the neurons of the substantia nigra and ventral tegmental areas of the brain. Hyaline inclusion bodies that resembled Lewy bodies were found in the neurons of mice with head shake 1 month after infection. Therefore, mice infected with N. asteroides may serve as a model for studying parkinsonian signs and other degenerative diseases involving extrapyramidal and pyramidal systems.

An ultrastructural study of HeLa cell invasion with Salmonella typhi GIFU 10007.

Scanning electron micrograph of HeLa S3 monolayered cells, inoculated with viable bacteria of a Salmonella typhi strain GIFU 10007, revealed that the extended microvilli tangled the bacteria within 10 min after inoculation. The micrographs of HeLa cells, at 1 hr after inoculation, indicate the following: shortening of bacterium-attached microvilli, subsiding of tangled bacteria into microvilli bush, and then attachment of bacterial soma to cell surface making the cell membrane depressed. The transmission electron micrographs, at 1 hr after inoculation, demonstrated the findings of interaction between HeLa cell and S. typhi 10007, similar to those observed on scanning electron micrographs. Hair-like fine structures from the soma of challenge organisms were also observed. They were in contact with HeLa cell microvilli and cell membrane. The bacteria were first partially and then totally surrounded by the HeLa cell plasma membrane. One, two, or several bacteria with intact outer membrane were enclosed in intracytoplasmic membrane-bound vacuoles. Fragmented vacuolar membrane was still visible around the intracellularly accumulated bacteria at 24 hr after inoculation. The viable cells of S. typhi 10007 are regarded as internalizing into HeLa cells by a process of endocytosis and to multiply within the membrane-bound vacuoles.

Cytopathogenic effect of Salmonella typhi GIFU 10007 on M cells of murine ileal Peyer's patches in ligated ileal loops: an ultrastructural study.

An electron microscopic study revealed that, within 30 min after inoculation into the ligated ileal loop of anesthetized mice, cells of Salmonella typhi GIFU 10007 adhered to the M cell surface of Peyer's patch lymphoid follicle epithelium, and induced almost complete destruction of M cells. The M cell cytoplasms were pinched off and extruded from the epithelial lining into the luminal space together with the lymphoid cells primarily enfolded into the corresponding M cells. When two or more M cells were destroyed, a large defect in the epithelial lining was apparent, and a number of bacteria appeared near the basal lamina of the epithelial lining. These findings suggest, as far as anesthetized murine ileal loops and strain 10007 are concerned, that ileal M cells are the target cell at an early stage of S. typhi infection and the infection may further progress to deeper tissues and to the general circulation.

Lactose-fermenting, multiple drug-resistant Salmonella typhi strains isolated from a patient with postoperative typhoid fever.

Two lactose-fermenting Salmonella typhi strains were isolated from bile and blood specimens of a typhoid fever patient who underwent a cholecystectomy due to cholelithiasis. One lactose-fermenting S. typhi strain was also isolated from a pus specimen which was obtained at the tip of the T-shaped tube withdrawn from the operative wound of the common bile duct of the patient. These three lactose-fermenting isolates: GIFU 11924 from bile, GIFU 11926 from pus, and GIFU 11927 from blood, were phenotypically identical to the type strain (GIFU 11801 = ATCC 19430 = NCTC 8385) of S. typhi, except that the three strains fermented lactose and failed to blacken the butt of Kligler iron agar or triple sugar iron agar medium. All three lactose-fermenting strains were resistant to chloramphenicol, ampicillin, sulfomethoxazole, trimethoprim, gentamicin, cephaloridine, and four other antimicrobial agents. The type strain was uniformly susceptible to these 10 drugs. The strain GIFU 11925, a lactose-negative dissociant from strain GIFU 11926, was also susceptible to these drugs, with the sole exception of chloramphenicol (minimal inhibitory concentration, 100 micrograms/ml).

[Antibacterial activity of cefoperazone against anaerobic bacteria (author's transl)].

The following results were obtained from the comparison of cefoperazone (CPZ) with cefazolin (CEZ), cephalothin (CET) and cefoxitin (CFX) in respect to their antibacterial action against anaerobic bacteria: 1) CPZ showed an antibacterial spectrum which was comparable to those of the known antibiotics, CEZ, CET and CFX. CPZ was particularly effective against anaerobic Gram-positive bacteria. 2) CPZ showed a distribution of susceptibility against clinically isolated strains which was comparable to those of CEZ, CET and CFX, CPZ was slightly inferior to CFX against B. fragilis and B. thetaiotaomicron, but was somewhat superior to any of CEZ, CET and CFX against anaerobic Gram-positive rods and B. distasonis. 3) The susceptibility of B. fragilis and F. necrophorum to CPZ decreased with an increase in the amount of inoculation, while C. perfringens and P. asaccharolyticus did not show any difference in susceptibility with the amount of inoculation. 4) The values of MIC and MBC of CPZ showed good coincidence with each other, thereby testifying its superior bactericidal action. 5) Natural resistant mutants were not obtained. 6) According to the examination of resistance in a test tube, P. variabilis did not show any appreciable increase in resistance, but F. varium increased its resistance rapidly. 7) CPZ was inferior only to CFX in stability against the beta-lactamase of B. fragilis. All of CEZ, CET and CMD lacked stability against the beta-lactamase of B. fragilis. 8) CPZ was found effective for the treatment of experimental subcutaneous abscess in mice caused by F. necrophorum.