Of 29 hematopoietic cell lines tested for susceptibility to human immunodeficiency virus (HIV)-1HTLV-IIIB infection, all CD4+ cell lines became infected. Continuous culturing of infected cell lines resulted in nine HIV-1 carrier cell lines, including, for the first time, an HIV-1 carrier megakaryoblastic cell line, MEG-01/HIV. The immunophenotypic profiles of a total of 17 HIV-1 carrier cell lines (nine newly and eight previously established cell lines) were compared with their respective parental noninfected cell lines. Except for total absence of CD4 expression, the expression of other antigens was variable among the 17 HIV-1 carrier cell lines. Persistent and consistent replication of infectious HIV-1 was detected in all of them in varying quantities. The great variability observed in both the altered marker expression, with respect to that of the noninfected parental cell lines, and in the quantities of persistently produced infectious HIV-1 was, nevertheless, specific to the individual cell lines. Furthermore, the present study demonstrates that there is no apparent correlation in the quantity of HIV-1 produced to either T cell, myelomonocytic cell, or megakaryocytic cell types. Instead, the results suggest that a particular interaction between HIV-1 and individual clonal cell lines may provide insight into the extremely complex immune dysregulation associated with the pathogenesis of acquired immune deficiency syndrome. Thus, the 17 HIV-1 carrier cell lines of diverse origin presented here provide valuable and unique models for further understanding acquired immune deficiency syndrome pathogenesis at the cellular and molecular levels. [P.S.E.B.M. 1993, Vol 202]
The stability of Δ9-tetrahydrocannabinol acetate (Δ9-THC-OAc) in the presence of diluents or during vaping was investigated. Laboratory-made pure Δ9-THC-OAc and glycerol, propylene glycol (PG), or polyethylene glycol 400 (PEG400) were dissolved in ethanol and subjected to the quantification method for Δ9-tetrahydrocannabinol (Δ9-THC) recommended by UNODC. The pure Δ9-THC-OAc diluted by glycerol, PG, or PEG400 was stored at 40°C for 27 days. A retail liquid product of Δ9-THC-OAc containing Δ9-THC as an impurity was stored at 80°C for 28 days in the presence of glycerol, PG, or PEG400. A cartridge containing the Δ9-THC-OAc liquid was installed in an e-cigarette device, which was heated and vaporized using a syringe connected to the cartridge through a silicone tube. The Δ9-THC-OAc liquid was put into a test tube and heated at 200 or 400°C for 5 min. The relative concentrations of cannabinoids in the test solution before and after each operation were measured by liquid chromatography with photodiode array detection or by gas chromatography with flame ionization detection. Δ9-THC was not detected in any cases of pure Δ9-THC-OAc, and it decreased or disappeared in all cases of Δ9-THC-OAc liquid. The present result showed that Δ9-THC-OAc does not decompose into Δ9-THC during each process.
Introduction: Mitochondrial transcriptional factor A (TFAM), a nucleoid protein of mitochondrial DNA (mtDNA), is essential not only for transcription and replication but also maintenance of mtDNA. We have previously reported that, in failing myocardium, TFAM expression and the myocardial mtDNA copy number were decreased. Moreover, in myocardial infarction, overexpression of TFAM attenuated the decrease in the mtDNA copy number, ameliorated cardiac remodeling, and markedly improved survival. Since exogenously-administrated recombinant TFAM protein has been shown to enter mitochondria in cultured cells, we examined our hypothesis that recombinant TFAM protein administration attenuates pathological remodeling in cardiac myocytes. Methods and Results: We prepared recombinant human TFAM including mitochondrial targeting signal by glutathione s-transferase fusion protein purification protocol. Recombinant TFAM was successfully recruited into mitochondria of rat neonatal cardiac myocytes, whereas TFAM without mitochondrial targeting signal was observed in nucleus. Treatment with TFAM increased the mtDNA copy number dose-dependently (100 nM: 1.8±0.1-folds), whereas it did not change morphology or population of mitochondria (electron microscopy). To elucidate the mechanism of TFAM on signaling pathways in cardiac hypertrophy, we investigated the effects of TFAM on the nuclear factor of activated T cell (NFAT) signaling, which is a major transcriptional factor regulating pathological hypertrophy and remodeling. TFAM totally abolished NFAT nuclear translocation induced by both angiotensin II (41±3% to 2±1%, p<0.01) and endothelin-1 (43±3% to 3±1%, p<0.01), suppressing respective NFAT transcriptional activity and NFAT-dependent gene expression. TFAM inhibited subsequent morphological hypertrophy of cardiac myocytes (angiotensin II: 1686±37 to 1167±20 μ m 2 , p<0.01, and endothelin-1: 1759±44 to 1206±21 μ m 2 , p<0.01). Conclusion: Recombinant TFAM increases the mtDNA copy number, and attenuates angiotensin II and endothelin-1-induced hypertrophy via inhibiting NFAT signaling. Recombinant TFAM would be an attractive, novel therapeutic strategy for cardiac hypertrophy and remodeling.
A highly sensitive ELISA for the determination of polymyxin B sulfate (PMB) was developed which is capable of measuring as low as 32pg/ml. Anti-PMB antibody was obtained by immunizing rabbits wiht PMB conjugated with mercaptosuccinyl bovine serum albumin (MS. BSA) using N-(γ-maleimidobutyryloxy) succinimide (GMBS) as a heterobifunctional coupling agent. An enzyme marker was similarly prepared by coupling PMB with horseradish peroxidase (HRP) employing GMBS. This ELISA showed very low reactivity with the PMB analogue, polymyxin E (0.05%). The values for PMB concentration detected by this assay were comparable with those detected by the bioassay. Moreover, the ELISA was about 10000 times more sensitive in detecting PMB at lower concentrations. Serum PMB concentration after the oral administration of a PMB tablet to human subjects was determined by the ELISA. PMB was rapidly absorbed from the gastrointestinal tract after the administration. then slowly decreased. These results indicate that the ELISA may be a valuable tool for studies of the pharmacokinetics and pharmacodynamics of the anti-endotoxin drug, PMB.
Mucosa-associated lymphoid tissue (MALT) lymphomas are localized primarily in the gastrointestinal tract and are characterized by an indolent nature and favorable outcome with specific therapy. Gastric MALT lymphomas are closely linked to Helicobacter pylori (H. pylori) infection, for which eradication therapy is recognized as an effective primary treatment for the disease. However, there is little information about long-term outcomes after the therapy. In the present study, we elucidated the long-term outcomes of 74 patients (70 H. pylori-positive and 4 negative cases) followed up by endoscopy at least 12 months after exclusive eradication therapy alone. The median follow-up period was 46 months. When the remission status was estimated at the time point of 12 months post-eradication, the numbers of patients with complete remission (CR), histologically residual disease with macroscopic normalization (hRD), partial remission with more than 50% tumor reduction (PR) or no response (NR) were 56, 12, 2 and 4, respectively. During follow-ups of over 12 months post-eradication, 11 of the 12 hRD cases were belatedly induced to CR but one CR case histologically relapsed into hRD. One of the 2 PR cases eventually turned into hRD 20 months later. Therefore, 66 CR, 3 hRD, 1 PR, and 4 NR cases (including 3 H. pylori-negative) were identified at the last follow-up of the present study. All 74 patients were followed up without any second-line therapies, but none exhibited disease progression. Thus, the long-term outcome of localized gastric MALT lymphoma after H. pylori eradication therapy was favorable. A watch and wait strategy may be a reasonable approach for hRD since the majority might be in the process of turning into delayed CR.
