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J B Hudson

Publications and source records attributed to J B Hudson.

At least 55 records · Page 3Linked to original sources

Plant photosensitizers with antiviral properties.

Many antiviral compounds obtained from plants are photosensitizers, i.e., their biological properties are dependent upon or augmented by light of specific wavelengths, commonly long wave ultraviolet, UVA. Three groups of chemically distinct plant photosensitizers have been investigated in some detail in regard to antiviral properties. These are (a) thiophenes and polyacetylenes; (b) furyl compounds; (c) certain alkaloids. Some of the thiophenes and their acetylenic derivatives possess extremely potent phototoxic activities toward membrane-containing viruses. These activities are markedly affected by the chemical structures of these compounds. Inactivated virus retains its integrity, however, and penetrates cells, but does not replicate. Their mechanism of action is believed to occur via singlet-oxygen damage to the membranes, although other targets cannot be ruled out. In contrast, the antiviral activities of plant furyl compounds (such as psoralens and furanochromones) appear to depend on UVA-mediated covalent adduct formation with the viral genomes. Some of the photoactive beta-carboline alkaloids also have impressive antiviral activities, especially against viruses with single-stranded genomes. These and other types of alkaloids appear to work by mechanisms that do not require covalent bonding to nucleic acids, and may also involve other target molecules as well. Some of these compounds have potent antiviral activities at concentrations well below cytotoxic levels, and accordingly should be tested in animal models.

Alkaloids↗

The antiviral action of lignans.

A total of 18 purified lignans was evaluated for antiviral activity against murine cytomegalovirus (CMV) and Sindbis virus, by means of different treatment regimens. Podophyllotoxin and alpha-peltatin were the most potent compounds, and they apparently inhibited murine CMV at an essential early step in the replication cycle after the adsorption of virus to the cells. On the other hand, justicidin B and the diphyllin derivatives were much more effective against Sindbis virus, and 12 of the lignans had no demonstrable effect at all, despite their known activities in other bioassays.

Antiviral Agents↗

The DNA-binding properties of immediate-early and early proteins of murine cytomegalovirus.

The DNA-binding properties of the immediate early (IE) and early proteins induced by the murine cytomegalovirus (Smith strain) in permissively infected 3T3-L 1 cells and nonpermissively infected J774A.1 cells were characterized. Eight of 10 IE proteins and 4 of 7 early proteins had affinity for DNA (native or denatured). A 96K IE protein was observed to have affinity for denatured DNA-cellulose when synthesized in J774A.1 cells but not when synthesized 3T3-L 1 cells.

Antigens, Viral↗

Analysis in vitro of two biologically distinct strains of murine cytomegalovirus.

The two biologically distinct strains of murine cytomegalovirus, "Smith" and "K181", showed slightly different restriction-endonuclease profiles; but we could not detect strain differences in the numbers or sizes of the major immediate-early, early or late viral proteins. It is possible however that some of the minor late proteins may differ.

Antigens, Viral↗

Further characterization of the murine cytomegalovirus induced early proteins in permissive and nonpermissive cells.

Some of the properties of the immediate-early (IE) and early proteins induced by the murine cytomegalovirus (Smith strain) were examined in permissively infected 3 T 3-L 1 cells, and in non-permissively infected J 774A.1 (mouse macrophage) and human fibroblast cells, in order to determine differences that could account for the restriction in virus replication in the latter two cell lines. The different virus induced proteins had distinctive partitioning characteristics between nuclear and cytoplasmic fractions. The 96 K major IE protein had an exclusively nuclear association, as did the most abundant early proteins of 39 K and 36 K. The other viral proteins however were evenly distributed between nucleus and cytoplasm. In general these patterns were also seen in the infected non-permissive cells. Several proteins showed more than one charge isomer on two-dimension gels, and in addition five IE proteins and two early proteins were phosphorylated. Only two differences between the permissive and nonpermissive infections were observed; the IE proteins of 100 K and 89 K when synthesized in the human cells had a stronger affinity for the nuclear fraction; also a phosphorylated form of the 30 K IE protein was not detected in MCMV infected J 774 A.1 cells.

Antigens, Viral↗

Further studies on the mechanism of centrifugal enhancement of cytomegalovirus infectivity.

The phenomenon of centrifugal enhancement of infectivity, previously shown to be an intrinsic property of murine and human cytomegalovirus (CMV) was studied further in an attempt to understand the underlying mechanism of action and to try to influence the phenomenon by means of extrinsic factors. The sand-rat herpes virus also displayed centrifugal enhancement in rat, mouse, and human cells. Further investigations were performed with murine CMV. The property of enhancement was not restricted to a specific phase of cell cycle. A variety of factors, known to influence cellular physiology, did not significantly affect the ratio of centrifugal infectivity to standard infectivity i.e., the c/s ratio. However, it was necessary to centrifuge the infected cultures within one hour of the establishment of infection, since centrifugation after this delay failed to yield the enhancement; but there was no partial effect; the cultures either showed complete centrifugal enhancement or no such effect. Thus, it appears that some herpes viruses, during their initial encounter with susceptible cells, can be forced into preferential productive infection by the process of centrifugation, but after a lapse of an hour or more, centrifugation can no longer do this and most of the infected cells fail to replicate virus. Consequently we continue to recommend low speed centrifugation (1000 x g, for 30 min) in order to enhance CMV detection and to optimize infectivity assays.

Animals↗

The efficacy of amantadine and other antiviral compounds against two salmonid viruses in vitro.

Various compounds, with known clinical efficacy against human viruses, were evaluated for their ability to inhibit the growth of infectious hematopoietic necrosis virus (IHNV, a rhabdovirus), and infectious pancreatic necrosis virus (IPNV, a birnavirus), in rainbow trout cell cultures. Amantadine inhibited the plaque forming ability of IHNV, at concentrations which did not affect cell growth or morphology, although it was not active against IPNV. Metisazone and bis-benzimidazole were also effective against IHNV; but they were slightly cytotoxic. Ribavirin, as expected, was active against IPNV, but was also equally effective against IHNV, although it was cytotoxic. Several other compounds were also tested but they were not inhibitory to either virus. The attraction of amantadine is the fact that relatively easy administration should be feasible.

Amantadine↗

Development of DNA probes for Candida albicans.

An attempt was made to produce DNA probes that could be used as a rapid and efficient means of detecting candidiasis (invasive Candida infection) in immunocompromised patients. Whole DNA from Candida albicans was digested with restriction endonuclease, and the resulting fragments were randomly cloned into a plasmid vector. Several recombinant plasmids were evaluated for cross-hybridization to various other Candida species, other fungal DNAs, and to nonfungal DNAs. Cross reactions were observed between the probes and different yeasts, but none with unrelated DNAs. Some recombinants were genus-specific, and two of these were applied to the analysis of C. albicans growth curves. It became evident that, although both 32P- and biotin-labelled probes could be made quite sensitive, a possible limitation in their diagnostic potential was the poor liberation of Candida DNA from cells. Thus, better methods of treatment of clinical specimens will be required before such probes will be useful in routine diagnosis.

Autoradiography↗

Characterization of a murine cytomegalovirus-induced immunosuppressive factor.

Murine cytomegalovirus infection in spleen cultures resulted in the production of a soluble factor, VISF (virus-induced suppressive factor), which inhibited concanavalin A mitogenesis in fresh spleen cells. Its production was specific for MCMV, since infection of spleen cultures by Sindbis virus, or bacteriophages PM 2 and T 4, and the phagocytosis of latex beads, all failed to elicit VISF. Maximum appearance of the factor occurred within 24 hours p.i. in spleen cultures, and its source was identified as the population of spleen cells which adhered to a plastic culture dish within two hours at 37 degrees C. Non-adherent cells did not produce the factor. Its production was not inhibited by indomethacin. VISF could be concentrated by ultrafiltration on a YM 2 membrane filter, and it was readily fractionated by chromatography on sephadex G-25. In relation to peptides of known molecular weight it appeared to be smaller than 1,400 daltons. Its ability to suppress concanavalin A mitogenesis was largely removed by digestion with proteinase K. Thus VISF appears to be a relatively small peptide or peptide-containing substance. It was purified further by HPLC.

Animals↗

Potentially useful antimicrobial and antiviral phototoxins from plants.

The wide range of naturally occurring compounds from microorganisms and plants which are phototoxic in UV-A light (320-400 nm) includes cinnamyl esters, coumarins and furanocoumarins, furanochromones, benzofurans, alkaloids, based on tryptophan or phenylalanine, extended naptho-and anthraquinones, polyacetylenes and their thiophene derivatives. The cellular targets are cell membranes, e.g., acetylenes, or the nucleus, e.g., furanocoumarins. Compounds reacting with cell membranes in light either generate singlet oxygen, e.g., alpha-terthienyl, or react by a free radical mechanism, e.g., phenylheptatriyne, or do both. Those that react with nucleic acids intercalate with these macromolecules forming photoadducts, e.g., furanocoumarins, furanochromones and furanoquinolines. With others, such as the beta-carbolines, the explanation for their photogenotoxicity is unknown. A number of these natural photosensitizers have been examined with bacteria, yeasts, and viruses and the mechanisms of phototoxicity have been elucidated. Some of the sulfur-containing acetylenes may be useful in photochemotherapy.

Anti-Bacterial Agents↗

Clinical significance of virulence-related assay of Yersinia species.

During the 42-month period from June 1982 through December 1985, 215 fecal specimens from 171 patients were found to be positive for yersiniae by using a combination of CIN agar and cold enrichment. Isolates were tested for markers of virulence including carriage of a plasmid 42 megadaltons in size, calcium dependence, autoagglutination, Congo red uptake, pyrazinamidase activity, fermentation of salicin, and hydrolysis of esculin. The results were correlated to symptoms in patients. A total of 80 Yersinia enterocolitica and 52 Y. enterocolitica-like strains (42 Y. frederiksenii, 8 Y. intermedia, and 2 Y. kristensenii) were examined. Positive virulence-related tests were as follows (for Y. enterocolitica, Y. frederiksenii, Y. intermedia, and Y. kristensenii, respectively): pyrazinamidase negativity, 12.5, 0, 0, and 50%; Congo red positivity, 5, 7.1, 87.5 and 0%; calcium dependence, 3.8, 0, 0, and 0%; autoagglutination positivity, 8.8, 0, 0, and 0%; carriage of the 42-megadalton plasmid, 28.6, 73.2, 5.7, and 0; salicin and esculin negativity, 12.5, 0, 0, and 50%. The isolates recovered from symptomatic patients were characterized in relation to the presenting symptoms. Isolates from 12 of 32 (37.5%) patients with acute-onset diarrhea and 9 of 30 (30.0%) patients with chronic symptoms expressed at least one virulence feature. No individual test or group of tests was consistently associated with onset or either type of symptoms. Routine testing of plasmid carriage, uptake of Congo red, calcium dependence, autoagglutination, and pyrazinamidase activity did not appear to provide information that would link the presence of symptoms with the virulence potential of fecal isolates of yersiniae.

Acute Disease↗

Functional changes in murine macrophages infected with cytomegalovirus relate to H-2-determined sensitivity to infection.

Peritoneal macrophages were infected with murine cytomegalovirus in vitro, and indices of infection and macrophage function were monitored over 4 days. When the cells were assessed for the expression of viral antigen or for cytopathic effects, infection was found to be solely determined by the H-2 phenotype. Less than 10% of the macrophages from resistant H-2k strains were affected, whereas 90% of H-2d cells and approximately 80% of H-2b and H-2a cells became infected. Similar trends were demonstrable by the measurement of viral DNA. In H-2a cells (B10.A), Dd conferred sensitivity despite the resistant K and class II phenotype. The findings suggest a critical association between the class I antigens and an early stage in the infectious process. Indices of infection were paralleled by a loss of Fc receptor expression and optimal colloidal gold uptake, whereas most cells remained trypan blue negative, retained dehydrogenase and acid phosphatase activities, and did not release infectious virus during the period of study. This is consistent with a role for macrophages in the persistence of cytomegalovirus in the host.

Animals↗