The serial cultivation of human diploid cell strains.

The limited in vitro lifetime of human diploid cell strains

The discovery that mammalian cells have the ability to synthesize the free radical nitric oxide (NO) has stimulated an extraordinary impetus for scientific research in all the fields of biology and medicine. Since its early description as an endothelial-derived relaxing factor, NO has emerged as a fundamental signaling device regulating virtually every critical cellular function, as well as a potent mediator of cellular damage in a wide range of conditions. Recent evidence indicates that most of the cytotoxicity attributed to NO is rather due to peroxynitrite, produced from the diffusion-controlled reaction between NO and another free radical, the superoxide anion. Peroxynitrite interacts with lipids, DNA, and proteins via direct oxidative reactions or via indirect, radical-mediated mechanisms. These reactions trigger cellular responses ranging from subtle modulations of cell signaling to overwhelming oxidative injury, committing cells to necrosis or apoptosis. In vivo, peroxynitrite generation represents a crucial pathogenic mechanism in conditions such as stroke, myocardial infarction, chronic heart failure, diabetes, circulatory shock, chronic inflammatory diseases, cancer, and neurodegenerative disorders. Hence, novel pharmacological strategies aimed at removing peroxynitrite might represent powerful therapeutic tools in the future. Evidence supporting these novel roles of NO and peroxynitrite is presented in detail in this review.

/pdf/nitric-oxide-and-peroxynitrite-in-health-and-disease-3uuuqgfdcz.pdf

Nitric Oxide and Peroxynitrite in Health and Disease

▪ Abstract At the interface between the innate and adaptive immune systems lies the high-output isoform of nitric oxide synthase (NOS2 or iNOS). This remarkable molecular machine requires at least 17 binding reactions to assemble a functional dimer. Sustained catalysis results from the ability of NOS2 to attach calmodulin without dependence on elevated Ca2+. Expression of NOS2 in macrophages is controlled by cytokines and microbial products, primarily by transcriptional induction. NOS2 has been documented in macrophages from human, horse, cow, goat, sheep, rat, mouse, and chicken. Human NOS2 is most readily observed in monocytes or macrophages from patients with infectious or inflammatory diseases. Sustained production of NO endows macrophages with cytostatic or cytotoxic activity against viruses, bacteria, fungi, protozoa, helminths, and tumor cells. The antimicrobial and cytotoxic actions of NO are enhanced by other macrophage products such as acid, glutathione, cysteine, hydrogen peroxide, or superoxid...

Nitric oxide and macrophage function

http://theory.bio.uu.nl/immbio/Marks91.pdf

By-passing immunization: Human antibodies from V-gene libraries displayed on phage

OBITUARY: Albert B. Sabin (1906-1993)

Pharmaceutical compositions that comprise a pharmaceutically acceptable carrier or diluent and a antibodies that each comprise at least a portion of the variable region from MAb 1112-1 are disclosed. Methods of treating individuals suspected of exposure to rabies are disclosed.

Methods for treating post-exposure rabies and anti-rabies compositions

Chicken-embryo vaccine.

SummaryThe reproductive capacity of several type 3 poliovirus strains was studied at 23°C, 37°C, and 40°C. There was marked variation in the capacity of the strains to replicate at different temperatures. Two wild strains, L-97 and L-38, were serially passed at 23°C. After 14–16 passages at 23°C, each strain lost its growth potential at 40°C while its growth potential at 23°C increased. A plaque pool obtained from the L-97 strain had the interesting ability to replicate well at all 3 temperatures. This plaque pool did not entirely lose its growth potential at 40°C even after 16 passages at 23°C. A plaque obtained from L-38 was passed at 23°C. There was a marked alteration in reproductive capacity at various temperatures of this particular plaque pool during a single passage, the third, at 23°C.

Effect of Cold Passage on Reproductive Capacity at Various Temperatures of Several Type 3 Poliovirus Strains.

A paper entitled “Pathogenesis of Alfalfa mosaic virus in Soybean (Glycine max) and Expression of Chimeric Rabies Peptide in Virus-Infected Soybean Plants” by Nina Fleysh, Deepali Deka, Maria Drath, Hilary Koprowski, and Vidadi Yusibov was published recently in Phytopathology (1). Two claims were made in the paper: (i) Soybean can be successfully infected with wild Alfalfa mosaic virus (AIMV), and (ii) recombinant of AIMV coat protein fused to rabies virus peptide could infect soybean. In trying to extend the data presented in the paper, we were unable, using the same rabies construct, to confirm the expression of rabies epitope in the soybeans. We performed exact replicas of the described experiments to reexamine the results. The plasmid clones pUT17, pUT27, pALg24, and A4 (1) were from the laboratory collection. Plasmids were linearized with the restriction enzymes SmaI, PstI, PstI, and SmaI, respectively, and transcribed with the T7 RNA polymerase (pUT17, pUT27, and pALg24) or SP6 RNA polymerase (A4) in vitro in the presence of the cap-analogue as described (1). The four RNA transcripts were inoculated into soybean as described (1) and symptom development was monitored 1 to 4 weeks postinoculation; at least three plants were used per sample for inoculations. To confirm the status of the inoculated plants, leaf tissue was sampled at different times postinoculation and tested in immunoblots (1) using AlMVspecific antibodies (Agdia, Elkhart, IN). As a positive control for the quality of transcription, an NFlg24 clone of the AIMV RNA3 was processed in parallel, i.e., the NFlg24 clone was linearized with SmaI and used for transcription with T7 RNA polymerase in the presence of the cap-analogue. The resulting NFlg24 transcript was inoculated to P12 Nicotiana tabacum plants that can support the replication of the RNA3 of AIMV (Fig. 1A). The development of symptoms in P12 plants was monitored visually 1 to 3 weeks postinoculation and confirmed by immunoblotting (Fig. 1A). In control experiments, AIMV was inoculated into soybean plants as described and symptom development and virus accumulation was monitored 1 to 4 weeks postinoculation (Fig. 1B). The results of the testing of recombinant, capped transcripts in soybean and N. benthamiana plants are summarized in Table 1. The capped transcripts of pUT17, pUT27, pALg24, and A4 did not produce visible symptoms in soybean or N. benthamiana upon inoculation. The lack of productive infection was confirmed by western blots of the lower inoculated leaves as well as upper uninoculated leaves (Table 1). The control, capped transcript NFlg24 resulted in normal productive infection upon inoculation into the P12 tobacco plants with clearly visible local and systemic

https://apsjournals.apsnet.org/doi/pdfplus/10.1094/PHYTO.2002.92.12.1260

Pathogenesis of Alfalfa mosaic virus in Soybean (Glycine max) and Expression of Chimeric Rabies Peptide in Virus-Infected Soybean Plants-A Reexamination.

Hilary Koprowski

Papers

OBITUARY: Albert B. Sabin (1906-1993)

Methods for treating post-exposure rabies and anti-rabies compositions

Chicken-embryo vaccine.

Effect of Cold Passage on Reproductive Capacity at Various Temperatures of Several Type 3 Poliovirus Strains.

Pathogenesis of Alfalfa mosaic virus in Soybean (Glycine max) and Expression of Chimeric Rabies Peptide in Virus-Infected Soybean Plants-A Reexamination.