Methods for generating point mutations at predetermined sites of RNA or DNA genomes have been developed. With Qbeta RNA as a template, minus strands were synthesized in vitro in a stepwise, substrate-controlled reaction. The nucleotide analogue N4-hydorxyCMP was introduced in the desired position, the minus strands were completed with the four standard triphosphates and used as templates to synthesize plus strands; about 30% of the progeny plus strands showed a base transition at the position corresponding to the nucleotide analogue. Two mutant RNAs with extracistronic nucleotide substitutions have been generated; one of these was viable, albeit with a reduced propagation rate, while the other was non-infectious. Furthermore, mutants with changes at the initiation codon of the coat cistron were prepared. An analysis of ribosome binding to such mutant RNAs revealed the importance of the A-U-G region for the formation of the initiation complex. With a similar approach applied to the beta-globin complementary DNA (CDNA) plasmid PbetaG, point mutations have been introduced at the positions corresponding to amino acids 121 to 123.

Base Sequence↗

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Lack of expression of antigens for islet cell antibodies in rat fetal pancreas.

It is not clear yet when pancreatic islet cells begin to express antigens for islet cell antibodies (ICA). Therefore, we studied whether human ICA-positive sera, crossreacting with adult rat islet cells, react with fetal and neonatal rat islet cells immunohistologically. The ICA did not react with fetal islet cells. On the other hand, neonatal islet cells over 2-3 weeks of age expressed the reactivities almost similar to those of the adult. It was suspected that pancreatic islet cells started to express their antigens to induce ICA gradually after birth.

Aging↗

PubMedDate not supplied

Localization of three subtypes of Fc gamma receptors in human placenta by immunohistochemical analysis.

We investigated the localization of the three subtypes of (Fc gamma R) in the normal human placenta using immunohistochemical and immunocytochemical techniques with specific monoclonal antibodies. The analysis revealed that Fc gamma RI was expressed on Hofbauer cells, that Fc gamma RII was expressed on Hofbauer cells and endothelial cells of fetal vessels, while Fc gamma RIII was expressed on trophoblasts, especially syncytiotrophoblasts. Moreover, we demonstrated that the expression of Fc gamma RI and Fc gamma RII on Hofbauer cells and endothelial cells of the fetal vessels in the 1st trimester placenta was different from that in the 3rd trimester placenta. These results, therefore, indicate that the three subtypes of Fc gamma R in the human placenta may contribute to maintenance of placental functions, because each Fc gamma R molecule displays unique biological functions similar to those on leukocytes.

Animals↗

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Platelet protein kinase C levels in Alzheimer's disease.

Alzheimer's disease (AD) has been suggested to be a systemic disease, and signal transduction abnormalities have been reported in non-neuronal AD cells. We have previously quantified the protein kinase C (PKC) subtypes in AD and control brains using a two-site enzyme immunoassay (EIA), and have shown that type II PKC levels were significantly reduced in the temporal cortex of AD patients. In this study, we used this EIA to assess the platelet levels of type II PKC in age-matched groups of AD patients and normal controls. The cytosolic level of type II PKC was significantly higher in AD platelets than in control platelets but was unchanged in the membranous fraction. Platelet proteins showed no differences between the AD and control groups. Therefore, the type II PKC content of the cytosolic fraction was increased in AD platelets. These results suggest that type II PKC may be altered in both the brain and platelets of AD patients and support the hypothesis that AD is a systemic disease.

Aged↗

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Significance of chorioamnionitis.

In preterm deliveries, we have reported a high incidence (30-50%) of histologic chorioamnionitis (CAM) in the placenta. There is little evidence about the effects of CAM on preterm infants. We investigated the levels of complements and cytokines in the cord blood, the pathological nature of the placenta, the L/S ratio of gastric and tracheal aspirate of each preterm infant at birth, and assessed the biological effects of CAM on them. CAM stimulates the immunological system by cytokine production (IL6 and IL8) and complement activation in the fetus. It has been suggested that CAM may be one of the factor accelerating fetal maturation of the immunological system such as complement activation and immunoglobulin production, and of surfactant synthesis in the lung. On the contrary, CAM may damage the structures along the lining cells in the airway by accumulating polymorphonuclear cells of the infants with Wilson-Mikity syndrome.

Chorioamnionitis↗

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Prevalence of normal-tension glaucoma and primary open-angle glaucoma in patients with collagen diseases.

PURPOSE: To investigate the prevalence of normal-tension glaucoma (NTG) and primary open-angle glaucoma (POAG) in patients with collagen diseases and determine whether an immunocompromised condition is present in a subset of glaucoma patients. METHODS: Three glaucoma specialists prospectively examined patients with collagen diseases. The diagnostic process included applanation tonometry, slit-lamp examination, gonioscopy, direct ophthalmoscopy, and automated static perimetry. Twenty-four-hour intraocular pressure monitoring was done when necessary. Using the results of a population-based survey conducted in Japan, we calculated an expected number of cases of NTG and POAG, and compared these with the actual number of cases. RESULTS: Of the 153 patients with collagen diseases examined, we found 6 patients with NTG and 2 patients with POAG. Of these 8 patients, 2 with progressive systemic sclerosis (PSS), one with NTG, and the other, POAG, had a history of being on systemic steroidal therapy. The prevalence of NTG and POAG was significantly higher in women patients having collagen diseases as compared with normal women (P = .027). CONCLUSION: Women patients with collagen diseases are highly susceptible to NTG and POAG.

Adult↗

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Clinical isolates of Vibrio cholerae O1 not producing cholera toxin.

Explore the source record for details and available documents.

Cholera↗

PubMedDate not supplied

Recombinant cholera toxin B subunit acts as an adjuvant for the mucosal and systemic responses of mice to mucosally co-administered bovine serum albumin.

We examined the mucosal adjuvant activity of recombinant cholera toxin B subunit (rCTB) produced by Bacillus brevis carrying pNU212-CTB by intranasal or oral co-administration of bovine serum albumin (BSA). Intranasal administration stimulated a high level of BSA-specific serum IgG antibody response and BSA-specific IgA antibody responses in the nasal and pulmonary lavages. Oral administration induced a moderate level of BSA-specific serum IgG antibody and a low level of BSA-specific IgA antibody in the large intestinal washes. These results show that CTB alone can act as an intranasal or oral delivery carrier; it also has strong adjuvant properties for stimulating serum IgG and mucosal IgA immune responses to unrelated, non-coupled antigens after intranasal or oral co-immunization.

Adjuvants, Immunologic↗

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The role of interferon regulatory factors in the interferon system and cell growth control.

Complex cellular responses are often coordinated by a genetic regulatory network in which a given transcription factor controls the expression of a diverse set of target genes. Interferon regulatory factor (IRF)-1 and IRF-2 have originally been identified as a transcriptional activator and repressor, respectively, of the interferon-beta (IFN-beta) as well as of IFN-inducible genes. However, these factors have since been shown to modulate not only the cellular response to IFNs, but also cell growth, susceptibility to transformation by oncogenes, induction of apoptosis, and development of the T cell immune response. Furthermore, the evidence suggests that deletion and/or inactivation of the IRF-1 gene may be a critical step in the development of some human hematopoietic neoplasms. Subsequently, these factors have been shown to constitute a family of transcription factors, termed the IRF-family. Recent studies indicate that other IRF family members also involve the regulation of the IFN system and cell transformation. The IRF-family may be examples of transcription factors which can selectively modulate several sets of genes depending on the cell type and/or nature of the cellular stimuli, so as to evoke host defense mechanisms against infection and oncogenesis.

Animals↗

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The interferon system and interferon regulatory factor transcription factors -- studies from gene knockout mice.

Interferon regulatory factors (IRFs) were initially identified as regulators of IFN-alpha/beta genes and to date nine members have been determined in human and mouse. They share a conserved DNA-binding domain in the N-terminal portion that recognizes similar DNA sequences. Despite their similar DNA binding specificity, the IRFs show diverse functions in response to extra cellular stimuli. Although the study of IRFs was started with respect to regulation of the IFN-alpha/beta gene expression, recent studies have revealed other aspects of IRF functions. In this review, we summarize our current knowledge of the functions of IRF family members revealed by our gene targeting study in mice, focusing on the regulation of the IFN system.

Animals↗

PubMedDate not supplied

Structure and expression of a cloned cDNA for human interleukin-2.

A cDNA coding for human interleukin-2 (IL-2) has been cloned from a cDNA library prepared from partially purified IL-2 mRNA. The DNA sequence codes for a polypeptide which consists of 153 amino acids including a putative signal sequence. A biologically active polypeptide, characteristic of human IL-2, was produced when the cDNA was fused to a simian virus 40 promoter sequence and used to transfect cultured monkey COS cells.

Amino Acid Sequence↗