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Biomedical subjects

D Amsterdam

Publications and source records attributed to D Amsterdam.

12 recordsLinked to original sources

Simple inoculum standardizing system for antimicrobial disk susceptibility tests.

A collaborative study was undertaken to evaluate a simple, convenient device which expedites inoculum standardization for antimicrobial disk susceptibility tests. The Inocupac system (Minnesota Mining & Manufacturing [3M] Co., St. Paul, Minn.) was used to perform disk tests in parallel with the standard Bauer-Kirby method. Five investigators tested 100 selected isolates, each in triplicate. Inter- and intralaboratory precision of both disk procedures was essentially comparable. The Inocupac system failed to consistently provide satisfactory growth with some streptococci, but when testing common gram-negative bacilli, staphylococci, and most enterococci, the Inocupac system gave zones about the same size (+/-2 mm) as the Bauer-Kirby procedure. Interpretive agreement between the two test systems varied from 90 to 99% with different antimicrobial agents, and repeated tests with the Bauer-Kirby method demonstrated the same degree of interpretive agreement. The Inocupac system is a valid alternative method for inoculating disk susceptibility tests.

Anti-Bacterial Agents

Cytology, growth characteristics and cellular alterations following SV40-induced transformation of human foetal brain cells derived from a Gm2 gangliosidosis and control.

Cell cultures were derived from the cerebra of a control and a Gm2 gangliosidosis (Tay-Sachs disease (TSD)) foetus. Both cell lines were identified as astrocytic, based on the ultrastructural demonstration of glial fibres. The control culture exhibited morphological differentiation when exposed to dibutyryl cAMP, a finding which was not observed with the TSD cells. The TSD culture demonstrated the pathobiological features of the disease, which included the absence of hexosaminidase A, increased concentration of Gm2 ganglioside and the detection of membranous cytoplasmic bodies by electron microscopy. Control and TSD cells were exposed to SV40 virus, resulting ultimately in the isolation of transformants which differed from the parental cell types in morphology, growth rate and greatly accelerated cell death. Both control and TSD cell lines have been in propagation for over 200 subcultures and the transformants were identified as astrocytic, based on the retention of characteristic glial fibres. The control culture demonstrated a chromosome range of 34--63, with a mean of 47. In contrast, the TSD transformants exhibited a range of 50--107 and a mean of 74. Transformed lines retained their parental hexosaminidase isoenzyme profiles; Hex A and B in control, and Hex B in TSD cells. Membraneous cytoplasmic bodies persisted in the TSD line. Neither line could be induced to differentiate after exposure to cAMP. Additionally, they had a population doubling time of under 85 h and failed to release infectious virus particles. Significant alterations in the total quantity and distribution profile of gangliosides were noted following viral transformation. A large increase in the percentage of Gm3 and a more modest increase in Gm2 were detected. In contrast, transformed lines were characterized by substantial reduction in the percentage of glucosamine-containing Gm2 and polysialoganglioside. Additionally, cultures exhibited a characteristic reduction in ganglioside content after transformation. The in vitro transformation of human brain cells has resulted in the derivation of permanent astrocytic lines which are, by virtue of their rapid growth rate and long-term survival, uniquely suited and adapted to the large scale in vitro production of substantial quantities of cells required for extensive biochemical study. Significantly, those characteristics which are unique to the Gm2 gangliosidosis storage disease have been retained in a permanent model CNS cell line.

Brain

Light and electron microscopic studies of "nude" mice CNS after subcutaneous administration of the E variant of the encephalomyocarditis (EMC) virus.

Sixteen 3 month old "nude" mice, 24 of their litter mates and 30 Swiss mice were injected subcutaneously with 0.1 ml suspension of the E variant of the encephalomyocarditis (EMC) virus. While the mortality rate of the litter mates and Swiss mice during 5-7 days after inoculation was more than 40%, none of the "nude" mice died during the experiment. The surviving animals were sacrificed at 24 h intervals from day one to seven days after injection. Brain suspensions assayed for the presence of the virus yielded significant titers at 24 h in all groups, which increased during 7 days. The litter mates and Swiss mice showed proliferation of lymphocytes and microglial cells in the perivascular areas of the brain during the fifth to the seventh day. The "nude" mice, on the other hand, displayed no perivascular lymphocytic infilteration during the same periods. Ultrastructurally, all groups showed aggregates of ribosomes in the cytoplasmic matrix on the third day, which became enlarged in size on the 5th day. At 7 days, both litter mates and Swiss mice showed an increased number of necrotic cells, while these changes were not observed in the "nude" mice. These findings suggest that the high mortality rate in immunologically normal mice was related to the efforts of T cells to eliminate virus-infected cells and to produce extensive necrosis, while T cell-depleted animals showed good survival rates.

Animals

API computer profiles: correlation of API 20E with API 10S.

A comparison of identifications of 201 clinical isolates with the 21-test API 20 Enteric kit and the subset of tests in API 10S indicated 85.6% agreement at the species level and 93.5% agreement at the genus level.

Bacteriological Techniques

GM2 ganglioside in fetal Tay-Sachs disease brain cultures: a model system for the disease.

Cultured cells derived from Tay-Sachs disease (TSD) fetal cerebellum were shown to accumulate GM2 ganglioside when compared with control cultures. In contrast, fibroblasts derived from TSD fetal lung do not contain GM2. GM2 was identified by thin-layer chromatography (TLC) and confirmed by gas-liquid chromatography (GLC). Unlike fetal TSD brain, the cell cultures established from fetal TSD brain have high concentrations of globoside and GD3 and little or no asialo GM2(GA2). The GM2 and related glycosphingolipids in these cultured cells contain a high percentage of C24:0 and C24:1 fatty acids. In spite of these differences, this TSD cell strain is unique in that it accumulates GM2, and can therefore serve as a useful in vitro model for the study of TSD.

Cells, Cultured

Ultrastructural alterations of tissue cultures from human fetal brain infected with the E variant of EMC virus.

Explant outgrowths from human fetal brain were infected with 104 plaque forming units of the E variant of the encephalomyocarditis virus. Ultrastructurally, the majority of the cultured cells were astrocytes containing a moderate amount of glial fibrils. The earliest alterations at 44 hrs after infection of the culture consisted of dilatation of the rough endoplasmic reticulum (ER) and moderate enlargement of the mitochondria with increased density of their matrix. Twenty hours thereafter, increased amounts of the rough ER and of free ribosomes were observed in the infected cells. This was followed by aggregates of larger dense particles which developed into a parallel lattice-like pattern within the cytoplasm, presumably presenting viral particles, without obvious cytonecrosis. The present observations of the continuous development of the crystalloid formation of the virus within the cytoplasmic matrix as well as the mode of formation of the free ribosomes adjacent to the ER support the previous hypothesis that, after the uptake of the virus into the cytoplasmic matrix, it associates with the ER, where production of viral RNA polymerase and of viral RNA is initiated.

Astrocytes

Glucose tolerance and pancreatic ultrastructure in mice with long-term diabetes induced by EMC virus (M variant) (38518).

Of 27 young male CD-1 mice infected with the M variant of the encephalomycarditis (EMC) virus and surviving for 11 or 12 mo, all but three had normal glucose tolerance curves, and all displayed normal islet cell morphology, at the time of sacrifice, in spite of an initital hyperglycemic response. Three animals maintained diabetic glucose tolerance curves one year after infection but they, too, failed to show significant morphological alterations in their pancreatic beta cells.

Animals

EMC virus and cultured human fetal pancreatic cells. Ultrastructural observations.

Tissue cultures of pancreatic acinar cells of seven human fetuses were infected with either the M or the E variant of the encephalomyocarditis (EMC) virus. Between the third and fifth days after infection, severe ultrastructural damage was noted. Margination, condensation, and fragmentation of the nuclear chromatin and the appearance of numerous cytoplasmic vesicles filled with finely granular material were the major changes observed. Viral particles were not identified. One hundred plaque-forming units (PFU) were sufficient to induce these alterations with the M variant while 10,000 PFU were needed to obtain the same results with the EMC-E virus. The described lesions are qualitatively similar to those induced in vivo in experimental CD-1 mice infected with the virus.

Cell Nucleus