[Mechanism of HIV replication].
Explore the source record for details and available documents.
Gene Products, rev↗
Biomedical subjects
A Adachi
Publications and source records attributed to A Adachi.
Measurement of circadian rhythms of ocular melatonin in the pigeon by in vivo microdialysis.
Ocular melatonin rhythms were measured in pigeons (Columba livia) by in vivo microdialysis. The birds were placed in light-dark cycles with 12 h of light and 12 h of darkness (LD12:12) or continuous dim light (LLdim) after LD12:12. Under LD12:12, melatonin levels were low during the light and high during the dark. The rhythms persisted under LLdim with lower levels during the subjective day and higher levels during the subjective night. Two hours of light exposure in the middle of the night acutely suppressed melatonin to the daytime level. These results indicate that microdialysis is useful for studies investigating the mechanism regulating ocular melatonin rhythms.
Animals↗
Generation and characterization of a host cell-dependent gag gene mutant of human immunodeficiency virus type 1.
An in-frame gag gene mutant of human immunodeficiency virus type 1, which carries two amino acid substitutions in the center of the p24 coding region, was constructed in vitro, and its replication properties in several cell lines were examined. In CD4-negative SW480 cells transfected with the mutant clone, synthesis and processing of viral gag, pol, and env proteins occurred normally, and viral particles were produced. Virions derived from the transfection displayed a severe replication defect when inoculated into some CD4-positive cell lines (H9 and Molt4 clone 8), but in other lines (A3.01 and M8166), the mutant virus grew fairly well. The mutant was demonstrated to be defective at an early infection phase (from adsorption to integration) in Molt4 clone 8 cells but was normal in A3.01 cells. These results indicated that the Gag-p24 protein of human immunodeficiency virus type 1 plays an important role at the early infection phase in a cell-dependent manner.
Amino Acid Sequence↗
Rev-dependency of expression of human immunodeficiency virus type 1 gag and env genes.
Structural gene expression of human immunodeficiency virus type 1 (HIV-1) requires a viral regulatory protein, Rev transactivator. We investigated Rev-dependency of HIV-1 gene expression by various reporter systems. Expression of unspliced and single-spliced viral mRNAs was demonstrated to be differentially dependent on the Rev function. This difference of Rev-dependency was found not to be determined by cis-elements in gag, pol, and env coding sequences reported so far, and was lost when the reporter constructs containing minimum elements for Rev-responsiveness such as splice signals and rev responsive element were used for experiments. These findings indicated that the fundamental structure of HIV-1 mRNA was critical for the differential regulation of gene expression by Rev transactivator.
Animals↗
Glucose-responsive neurons in the brainstem.
AREA POSTREMA: The influence on feeding behavior caused by ablation of the area postrema (AP) in rodents indicates the participation of this structure in the control of ingestion. Two types of glucose responsive neurons were identified in the AP: one is characterized by increasing the discharge rate in response to glucose (glucoreceptor type) and the other by decreasing the discharge rates in response to glucose (glucose sensitive type). These glucose responsive neurons may participate in glycemic homeostasis. NUCLEUS OF SOLITARY TRACT: The glucose responsive neurons exist within the caudal portion of nucleus of the solitary tract (NTS), a relay station in visceral afferents. Two types similar to the AP were also recognized. It is confirmed that hepatic glucose sensitive afferents terminate on some of the glucose sensitive neurons. This convergence may serve as a fail-safe mechanism. In addition, the NTS involving complex neural networks of excitatory and inhibitory interneurons may be concerned with integration of glycemic information. DORSAL MOTOR NUCLEUS OF VAGUS: Some neurons within the dorsal motor nucleus of the vagus (DMV) were identified as the glucose responsive ones. Both types were also recognized. It is confirmed by antidromic activation that these glucose responsive DMV neurons send their axons toward the gastric or coeliac branch that innervates either the stomach, intestine or pancreas. Some of the DMV neurons may subserve an enteroceptor function by themselves. They may also play a role in the brainstem neural control of glycemic homeostasis as the fail-safe mechanism.
Animals↗
Comparison of trihalomethane in tap water with and without activated carbon adsorption during the preparation process of tap water.
Explore the source record for details and available documents.
Adsorption↗
Both SU and TM env proteins are responsible for monkey cell tropism of simian immunodeficiency virus SIV mac.
While simian immunodeficiency virus (SIV) derived from an infectious molecular clone pMA239 is tropic and pathogenic for monkeys, the virus derived from another infectious clone pMA142 does not replicate in monkey cells. To determine genetic sequences responsible for this tropism, a series of recombinant clones were constructed from pMA142 and pMA239. The determinant in pMA239 was mapped within regions encompassing the env gene. Viruses, which carry the 239 env gene encoding surface and/or transmembrane proteins, were tropic for monkey cells.
Animals↗
Functionality of chimeric Rev proteins of HIV/SIV.
Studies on functional compatibility of various Rev proteins derived from all known human and simian immunodeficiency virus subgroups have shown that this essential gene product is not always exchangeable among the viruses. In an attempt to map the region of Rev proteins responsible for the observed nonreciprocal complementation, hybrid genomic Rev expression vectors were constructed by exchanging the first and second exons of rev genes, and were examined for their abilities to activate reporter clones by transfection. With one exception, the second coding exon of rev gene determined the functional specificity of Rev proteins.
Animals↗
Functional analysis of human spuma retrovirus genome.
Human spuma retrovirus (HSRV) belongs to retroviruses that possess a complex genome organization. HSRV carries at least three extra genes in the region between env and the 3' long terminal repeat, which are not found in simple retroviruses. Via alternative splicing, these HSRV genes can encode several proteins. To genetically study the requirements of these viral proteins for viral replication in tissue cultures, a number of mutant viruses were constructed from an infectious molecular clone of HSRV. All mutants grew normally in the cell lines tested, except for those lacking transcriptional transactivator activity. By reporter-based transient assay systems, no Rev/Rex equivalent was detected in the HSRV proteins.
Alternative Splicing↗
Evaluation of gallbladder function before and after gastrectomy using a double-isotope method.
The motor function of the gallbladder after partial distal gastrectomy for gastric carcinoma (Billroth-I reconstruction) and its relationship to the motor function of the stomach was investigated using a double-isotope method. In this method, [99mTc]N-54-pyridoxyl-5-methyltryptophan ([99mTc]PMT) was utilized as a tracer for the biliary tract and [111In]diethyl-triaminopontacetic acid ([111In]DTPA) mixed with a liquid test meal was used as a tracer for the digestive tract. Gastric emptying half-time (GET1/2) was measured, since this was used historically as the physiologic indicator of gallbladder contractile stimulus. The volume of test meal that emptied from the stomach into the duodenum per minute (VOL/MIN) was measured, as was the gastric emptying duration (GED). A series of gallbladder emptying phenomena were analyzed using three different criteria: gallbladder emptying half-time (GBET1/2), retention rate of bile in the gallbladder (RR), and the start of gallbladder bile ejection into the duodenum after ingestion (TL: time lag). While GET1/2 was reduced (P < 0.01) and VOL/MIN was increased (P < 0.01) after gastrectomy, GED was shortened remarkably (P < 0.01). GBET1/2 showed no significant change, RR increased (P < 0.05), and TL was prolonged (P < 0.01). The postoperative dysfunction of the gallbladder was caused by the short and intense stimulus on the biliary tract during the duodenal phase and by intraoperative injury to the innervation of the gallbladder. In particular, the hepatic branch of the left vagus nerve was injured during the right paracardiac lymph node dissection performed as a component of the radical gastrectomy.
Adult↗
Chemosensitivity of neurons in the dorsal motor nucleus of the vagus that responded to portal infusion of hypertonic saline in rats.
Neural responses in the dorsal motor nucleus of the vagus (DMV) to topical administrations of sodium and portal infusions of hypertonic saline were investigated electrophysiologically by using multibarrel electrodes in anesthetized rats. Of 102 neurons that showed antidromic response to electrical stimulation of the ventral gastric vagus or the accessory celiac vagus, 51 neurons increased and 13 neurons decreased their discharge rates in response to the electrophoretic administration of sodium. The other 38 neurons did not respond to this stimulation. The portal infusion of hypertonic saline elicited neural responses of some DMV neurons whose axons are involved into either the ventral gastric or the accessory celiac vagus. Further, effects of the topical administration and the portal infusion of hypertonic saline were examined on 33 neurons. Typical response was characterized by an increase in discharge rate responding to both of the portal infusion and the topical administration. In conclusion, the DMV neurons receiving the afferent inputs from hepatoportal osmoreceptors may have an enteroceptor function detecting the change in osmotic pressure of their environment.
Animals↗
Function of human immunodeficiency virus type 1 Vpu protein in various cell types.
We evaluated the function of human immunodeficiency virus type 1 vpu gene in various cell lines. We established a highly sensitive system consisting of chloramphenicol acetyltransferase and reverse transcriptase assays and used it to monitor the effects of mutation of the vpu gene. In some cell lines, Vpu protein was not required at the early phase of viral replication but was important for efficient virion production. In these cells, the Vpu protein functioned effectively irrespective of the presence of intact env gene products. Likewise, CD4 gene expression had no effects on Vpu function. In the other cell lines tested, Vpu protein was not important for virion release, and the vpu mutant clone generated a normal level of progeny virions upon transfection.
3T3 Cells↗
Early replication block of human immunodeficiency virus type 1 in monkey cells.
The genetic and functional basis of the replication-defective nature of human immunodeficiency virus type 1 (HIV-1) in monkey cells was studied. By the generation and characterization of chimeras between HIV-1 and simian immunodeficiency virus, the sequence encompassing the 3' half of the long terminal repeat, gag and pol genes of HIV-1 was found to be responsible for the growth restriction. Early and late phases of HIV-1 replication in monkey cells were analysed in detail using several assay systems: transfection/coculture, transcomplementation between various proviral clones carrying the CAT gene and effector clones and evaluation of transcription and reverse transcription. All the data were consistent with the notion that HIV-1 replication is blocked at a very early stage(s) such as uncoating and/or reverse transcription in monkey cells.
Animals↗
Targeting of chrolamphenicol acetyltransferase to human immunodeficiency virus particles via Vpr and Vpx.
Vpr and Vpx are the auxiliary proteins of human immunodeficiency viruses (HIVs) selectively incorporated into mature viral particles. We showed that the bacterial chloramphenicol acetyltransferase (CAT) fused to the N-terminus of HIV-1 Vpr, HIV-2 Vpr, or HIV-2 Vpx was incorporated into mature virions in a type-selective manner. By using chimeric proteins between HIV-1 Vpr and HIV-2 Vpx, we found that the N-terminal side of these proteins was mainly important for type-selective virion incorporation. The C-terminal arginine-rich region of HIV-1 Vpr was also found to transport CAT fusion proteins into virions but without any type selectivity. Furthermore, the corresponding regions of HIV-2 Vpr and HIV-2 Vpx had no such activity. This region of HIV-1 Vpr may interact nonspecifically with viral genomic RNA. Collectively, Vpr and Vpx may provide a means to introduce foreign proteins and other molecules into HIV virions for therapeutic purposes.
Amino Acid Sequence↗
Molecular epidemiology of Staphylococcus spp. contamination in the ward environment: study on mecA and femA genes in methicillin-resistant strains.
Staphylococcus spp. were isolated from the ward environment and antibiotic susceptibility tests were performed. Twenty-nine strains out of 274 isolates were S. aureus, and 41.4% of the S. aureus strains were methicillin resistant (MRSA). All 12 strains of MRSA were also resistant to oxacillin, ceftizoxime, ampicillin and clindamycin. Among the coagulase-negative staphylococci (CNS), methicillin-resistant (MR) strains of S. epidermidis, S. capitis, S. warneri, S. haemolyticus, S. hominis, S. auricularis, S. saprophyticus and S. cohnii were isolated. Eight of the 10 S. Haemolyticus strains were methicillin resistant. The femA gene was detected in S. aureus (MSSA and MRSA), but not in CNS by polymerase chain reaction (PCR) analysis and Southern blot analysis. The mecA gene was found in all the MRSA and MR-S. epidermidis strains tested, and one of the two MR-S. hominis strains, but not in MSSA, MS-S. epidermidis, MS-S. hominis, or MS-S. haemolyticus. DNA from one strain of MR-S. hominis and 2 strains of MR-S. haemolyticus was not amplified by PCR using the mecA gene primer, or hybridized by Southern blotting. The ambiguity that mecA was detected in some MR-CNS strains, but not in others is discussed.
Base Sequence↗
Effects of translation initiation factor eIF-5A on the functioning of human T-cell leukemia virus type I Rex and human immunodeficiency virus Rev inhibited trans dominantly by a Rex mutant deficient in RNA binding.
The viral transactivator proteins Rex and Rev are necessary for the expression of structural proteins of human T-cell leukemia virus type I and human immunodeficiency virus type 1, respectively. Although the interaction of Rex/Rev with a cellular cofactor(s) has been thought to be required for Rex/Rev action, there is no suitable system to search for the cofactor(s) in mammalian cells. We found that a Rex mutant, TAgRex, which contains a simian virus 40 nuclear localization signal in place of the N-terminal 19 amino acids of Rex, could dominantly inhibit wild-type Rex/Rev functions. The inhibition did not require either Rev response element/Rex response element binding or the oligomerization ability of the mutant, but it did require a region around amino acid 90 of the Rex protein, suggesting that TAgRex sequestered the cellular cofactor. Complementation with the eukaryotic translation initiation factor 5A (eIF-5A) in this system could restore the impaired Rex function. These results indicate that eIF-5A is the cofactor indispensable for Rex function. Additionally, by using a two-hybrid system, the homo-oligomer formation of Rex was found to be mediated by the region around amino acid 90 in addition to Tyr-64 and Trp-65 of Rex protein. Thus, eIF-5A may play a part in the formation of the Rex homo-oligomer.
Animals↗
The CD5+ B cells and myasthenia gravis.
A high frequency of CD5+ B lymphocytes in the peripheral blood of patients with myasthenia gravis (MG) has been reported recently. These results seem to indicate an attractive linkage between CD5+ B lymphocytes and autoantibodies against Acetylcholine receptor in MG. We examined the frequency of CD5+ B cells in 20 patients with MG and 21 normal healthy controls by two-color flow cytometry. However, there were no significant differences in the percentages of CD5+ B lymphocytes between the two groups. We also examined the frequency of CD5+ B lymphocytes in the resected thymus of patients. The frequency of CD5+ B lymphocytes in the thymus was low and similar pattern to that in the peripheral blood. We checked the antibody (Ab) production against the human acetylcholine receptor in either CD5+ B or CD5- B lymphocytes using B lymphoblastoid cell line generated from the lymphocytes of 11 patients with anti-AChR Abs in the sera. Abs against the AChR in the human were mostly produced by CD5- B, not CD5+ B lymphocytes. The anti-AChR Abs (IgG) production of CD5+ B cells and CD5- B cells (mean +/- SD) were 6.8 +/- 2.4 fmol/ml and 18.5 +/- 17.6 fmol/ml, respectively. These results suggest that in MG, the frequencies of the CD5+ B lymphocytes in PBL may be genetic background and that there may be no strong linages between AChR Ab production and CD5+ B lymphocytes.
Adult↗
[Functions of human immunodeficiency virus regulatory genes].
Explore the source record for details and available documents.
Gene Expression Regulation, Viral↗