[Advance of molecular biological research of filamentous pathogenic fungi].
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Antifungal Agents↗
Biomedical subjects
L Han
Publications and source records attributed to L Han.
[Comparing Swerita daridi with Swertia mileencis on pharmacologic action].
The article compared Swertia daridi with Swertia mileencis about chemical composition and pharmacological action. The study showed two species drug's bitter taste chemical composition and pharmacological action are approximately equal. It can be tried to replace Swertia daridi with Swertia mileencis.
Animals↗
Fine structure of the murine leptin receptor gene: splice site suppression is required to form two alternatively spliced transcripts.
The fine structure of the murine leptin receptor gene (Lepr) is described. Duplicated ligand binding domains (conserved among cytokine receptors) are found in eight exons (coding exons 3 to 6 and 8 to 11). Thus, it is possible that a single leptin receptor molecule could have two functional ligand binding domains. The transmembrane region of Lepr is in coding exon 16 while the juxtamembrane JAK docking site is in coding exon 17. For all membrane-bound forms, the transcript must include 17 invariant exons and 1 alternatively spliced 3' terminal exon. The transcript encoding the soluble receptor (Re) includes 14 coding exons and an alternatively spliced 3' terminal exon. We have identified two splice variants (Rc and Re) for which there are no intervening sequences between the two final exons. This unusual juxtaposition of exons requires that splice donor sites at the 5' end of the respective terminal exons be ignored in the production of these splice variants. We suggest that splice site suppression is responsible for the formation of two of the alternatively spliced forms of the mouse Lepr gene. The juxtaposition of two coding exons separated by a consensus splice donor sequence is the structural substrate for this mode of alternative splicing. We present evidence that the Rc form is expressed in human tissues while the Re form, the soluble receptor, is not expressed.
Alternative Splicing↗
Protein binding and signaling properties of RIN1 suggest a unique effector function.
Human RIN1 was first characterized as a RAS binding protein based on the properties of its carboxyl-terminal domain. We now show that full-length RIN1 interacts with activated RAS in mammalian cells and defines a minimum region of 434 aa required for efficient RAS binding. RIN1 interacts with the "effector domain" of RAS and employs some RAS determinants that are common to, and others that are distinct from, those required for the binding of RAF1, a known RAS effector. The same domain of RIN1 that binds RAS also interacts with 14-3-3 proteins, extending the similarity between RIN1 and other RAS effectors. When expressed in mammalian cells, the RAS binding domain of RIN1 can act as a dominant negative signal transduction blocker. The amino-terminal domain of RIN1 contains a proline-rich sequence similar to consensus Src homology 3 (SH3) binding regions. This RIN1 sequence shows preferential binding to the ABL-SH3 domain in vitro. Moreover, the amino-terminal domain of RIN1 directly associates with, and is tyrosine phosphorylated by, c-ABL. In addition, RIN1 encodes a functional SH2 domain that has the potential to activate downstream signals. These data suggest that RIN1 is able to mediate multiple signals. A differential pattern of expression and alternate splicing indicate several levels of RIN1 regulation.
3T3 Cells↗
The packaging capacity of adeno-associated virus (AAV) and the potential for wild-type-plus AAV gene therapy vectors.
Because of its ability to integrate chromosomally and its non-pathogenic nature, adeno-associated virus (AAV) has significant potential as a human gene therapy vector. Here we investigate the maximum amount of DNA which can be inserted into the AAV genome and still allow efficient packaging into an infectious virus particle. Altered wild-type AAV genomes were constructed with inserts, which increased in size by 100 bp, ligated at map unit 96. These large wild-type-plus genomes were able to replicate and produce infectious virus, at levels slightly reduced but comparable to normal sized wild type, until the insert size reached 1 kb. These data indicate that the maximum effective packaging capacity of AAV is approximately 900 bp larger than wild type, or 119%. Furthermore, it is demonstrated that these large AAV genomes are able to latently infect cells by chromosomal integration as does wild-type AAV. These data suggest that therapy vectors carrying a foreign gene of 900 bp or less can be generated from AAV, by ligation into non-essential locations, and result in a recombinant AAV virus with a fully wild-type phenotype. Such wild-type-plus AAV vectors will have both advantages and disadvantages over defective recombinant AAV virus - the most important advantages being the ease in which high titers of infectious virus can be generated and the ability to specifically integrate within chromosome 19. Once the concern subsides over the presence of wild-type AAV in clinical applications, wild-type AAV vectors may find specific application niches for use in human gene therapy.
Cells, Cultured↗
Regulation of the oncogenic activity of BCR-ABL by a tightly bound substrate protein RIN1.
RIN1 was originally identified by its ability to physically bind to and interfere with activated Ras in yeast. Paradoxically, RIN1 potentiates the oncogenic activity of the BCR-ABL tyrosine kinase in hematopoietic cells and dramatically accelerates BCR-ABL-induced leukemias in mice. RIN1 rescues BCR-ABL mutants for transformation in a manner distinguishable from the cell cycle regulators c-Myc and cyclin D1 and the Ras connector Shc. These biological effects require tyrosine phosphorylation of RIN1 and binding of RIN1 to the Abl-SH2 and SH3 domains. RIN1 is tyrosine phosphorylated and is associated with BCR-ABL in human and murine leukemic cells. RIN1 exemplifies a new class of effector molecules dependent on the concerted action of the SH3, SH2, and catalytic domains of a cytoplasmic tyrosine kinase.
Adaptor Proteins, Signal Transducing↗
Human papillomavirus is more prevalent in first trimester spontaneously aborted products of conception compared to elective specimens.
In this study the possible role of human papillomaviruses (HPV) in spontaneous abortions is addressed by assaying for HPV DNA in first trimester spontaneous and electively aborted products of conception materials enriched for chorionic villi. The presence of HPVs was measured by polymerase chain reaction (PCR) amplification and DNA dot blot hybridization using an internal probe. The "broad spectrum" HPV primers were directed to amplify E6/E7 junction sequences, while the probe was of an HPV-16 sequence with significant homology to HPV-6/11. The quantity and quality of isolated DNA was also analyzed and compared by observing the PCR amplification of a cellular sequence from the human beta-globin gene. Fifteen of the 25 spontaneous samples (60%) were found to be positive for HPV E6/E7 sequences. In comparison, only 3 of the 15 elective samples (20%) were positive. This is the first study of HPV in fetal materials to incorporate material from elective abortions as a control group. Although confounding contamination from the cervix and vagina can't be ruled out, these data are significant and strongly suggest that HPVs are elevated in spontaneously aborted products of conception. Furthermore, these results suggest the possibility that HPVs may be etiologic agents of at least some spontaneous abortions.
Abortion, Induced↗
Nanophthalmos with longstanding choroidal effusion and serous retinal detachment.
PURPOSE: To report a typical case of nanophthalmos with uveal effusion and local serous retinal detachment followed for 1 year. METHODS: Clinical ocular examinations included vision, refraction, corneal diameter, anterior chamber depth, intraocular pressure, fundoscopy, A/B scan ultrasonography and gonioscopy. RESULTS: Both eyes were hypermetropic with small corneas and shallow anterior chambers. Decreased axial length and thickened sclera were also found. There were peripheral choroidal effusions and local serous retinal detachments as well. The patient declined any surgery that was offered. No significant change in either eye was found after 1 year follow-up. CONCLUSION: This case illustrates that the progress of choroidal effusion and retinal detachment in nanophthalmos may be very slow and even non-progressive for at least 1 year. In these cases sclerectomy and or sclerotomy may be delayed without undue immediate risk to the vision.
Anterior Chamber↗
In vivo and in vitro effects of thiolactomycin on fatty acid biosynthesis in Streptomyces collinus.
A stable-isotope assay was used to analyze the effectiveness of various perdeuterated short-chain acyl coenzyme A (acyl-CoA) compounds as starter units for straight- and branched-chain fatty acid biosynthesis in cell extracts of Streptomyces collinus. In these extracts perdeuterated isobutyryl-CoA was converted to isopalmitate (a branched-chain fatty acid), while butyryl-CoA was converted to palmitate (a straight-chain fatty acid). These observations are consistent with previous in vivo analyses of fatty acid biosynthesis in S. collinus, which suggested that butyryl-CoA and isobutyryl-CoA function as starter units for palmitate and isopalmitate biosynthesis, respectively. Additionally, in vitro analysis demonstrated that acetyl-CoA can function as a starter unit for palmitate biosynthesis. Palmitate biosynthesis and isopalmitate biosynthesis in these cell extracts were both effectively inhibited by thiolactomycin, a known type II fatty acid synthase inhibitor. In vivo experiments demonstrated that concentrations of thiolactomycin ranging from 0.1 to 0.2 mg/ml produced both a dramatic decrease in the cellular levels of branched-chain fatty acids and a surprising three- to fivefold increase in the cellular levels of the straight-chain fatty acids palmitate and myristate. Additional in vivo incorporation studies with perdeuterated butyrate suggested that, in accord with the in vitro studies, the biosynthesis of the palmitate from butyryl-CoA decreases in the presence of thiolactomycin. In contrast, in vivo incorporation studies with perdeuterated acetate demonstrated that the biosynthesis of palmitate from acetyl-CoA increases in the presence of thiolactomycin. These observations clearly demonstrate that isobutyryl-CoA is a starter unit for isopalmitate biosynthesis and that either acetyl-CoA or butyryl-CoA can be a starter unit for palmitate biosynthesis in S. collinus. However, the pathway for palmitate biosynthesis from acetyl-CoA is less sensitive to thiolactomycin, and it is suggested that the basis for this difference is in the initiation step.
Acetic Acid↗
A novel alternate anaplerotic pathway to the glyoxylate cycle in streptomycetes.
ccr encoding crotonyl coenzyme A (CoA) reductase (CCR), which catalyzes the conversion of crotonyl-CoA to butyryl-CoA in the presence of NADPH, was previously cloned from Streptomyces collinus. We now report that a complete open reading frame, designated meaA, is located downstream from ccr. The predicted gene product showed 35% identity with methylmalonyl-CoA mutases from various sources. In addition, the predicted amino acid sequences of S. collinus ccr and meaA exhibit strong similarity to that of adhA (43% identity), a putative alcohol dehydrogenase gene, and meaA (62% identity) of Methylobacterium extorquens, respectively. Both adhA and meaA are involved in the assimilation of C1 and C2 compounds in an unknown pathway in the isocitrate lyase (ICL)-negative Methylobacterium. We have demonstrated that S. collinus can grow with acetate as its sole carbon source even though there is no detectable ICL, suggesting that in this organism ccr and meaA may also be involved in a pathway for the assimilation of C2 compounds. Previous studies with streptomycetes provided a precedent for a pathway that initiates with the condensation of two acetyl-CoA molecules to form butyryl-CoA, which is then transformed to succinyl-CoA with two separate CoB12-mediated rearrangements and a series of oxidations. The biological functions of ccr and meaA in this process were investigated by gene disruption. A ccr-blocked mutant showed no detectable crotonyl-CoA reductase activity and, compared to the wild-type strain, exhibited dramatically reduced growth when acetate was the sole carbon source. An meaA-blocked mutant also exhibited reduced growth on acetate. However, both methylmalonyl-CoA mutase and isobutyryl-CoA mutase, which catalyze the two CoB12-dependent rearrangements in this proposed pathway, were shown to be present in the meaA-blocked mutant. These results suggested that both ccr and meaA are involved in a novel pathway for the growth of S. collinus when acetate is its sole carbon source.
Acetates↗
[Improvement of product technology on prepared soypean].
This paper reports the new prepared method of Prepared Soypean by purebred culture of Aspergillus niger. The technology condition is that temperture is 28 +/- 2 degrees C and comparative humidity is 95%, time of fermented culture is 15-20 days. After improvement, polluttion of other bacteria is easily avoided.
Aspergillus niger↗
Angiotensin-related induction of immediate early genes in rat brain.
Several studies are reviewed in which behavioral aspects of angiotensin (Ang) II on fluid intake have been compared with induction of the immediate early gene product, Fos, as a marker of neuronal activation in rat bain. Either peripheral or central administration of Ang II induced Fos along the lamina terminalis (SFO, MnPO, AV3V) and in the magnocellular neurosecretory groups (SO, PVH). A similar pattern is seen with central injection of renin. Both pharmacological and antisense oligonucleotide probe studies indicate that an AT1 receptor is involved, probably with the initial transduction in the SFO. Treatments that induce sodium appetite all induce Fos along the lamina terminalis, but usually not in the SO or PVN. Kininase II inhibitors, such as captopril, acutely potentiate drinking to Ang I, but after chronic exposure they may inhibit water intake. In contrast, the dipsogenic effect of bradykinin which is manifest in the presence of acute captopril remains unaffected by chronic administration. This suggests that the sodium appetite that appears with chronic captopril treatment may depend in part on peptides other than Ang.
Angiotensin II↗
Expression of Fos in rat brain in relation to sodium appetite: furosemide and cerebroventricular renin.
Two experiments were performed to investigate the relationship between the expression of sodium appetite and the appearance of Fos-like immunoreactivity (Fos-IR) in the brain of rats. In the first experiment, rats were depleted of sodium by treatment with furosemide 24 h prior to sacrifice and without access to either food or sodium solution. Some rats had access to distilled water, and others had no fluids available during the 24 h. All of the furosemide-treated rats showed Fos-IR in both the subfornical organ (SFO) and around the organum vasculosum laminae terminalis (OVLT). Rats with access to distilled water during the depletion period showed no Fos-IR in the supraoptic (SON) or paraventricular hypothalamic nuclei (PVN) and, in parallel behavioral studies, comparably-treated rats consumed only 0.3 M NaCl solution at the end of the 24 h. In rats that had no fluids during the deprivation period, only about one half showed Fos-IR in SON and PVN and, in parallel behavioral studies, comparably treated rats consumed both water and 0.3 M NaCl solution at the end of 24 h. In a second experiment, cerebroventricular administration of renin stimulated short latency intake of 0.3 M NaCl and water. The relative intakes of water and NaCl were comparable at a low dose of renin, but intake of water exceeded that of NaCl after higher doses. Renin induced Fos-IR in SFO, MnPO, peri-OVLT region, SON and PVN. Both Fos-IR and fluid intake were antagonized by administration of losartan, an angiotensin II type 1 receptor antagonist. Thus, only the circumventricular organs of the lamina terminalis showed Fos-IR during each natriorexigenic regimen in these studies. These data support the view that Ang II of both central and peripheral origin activates the SFO and/or peri-OVLT region and contributes to sodium appetite.
Animals↗
Dissociation of Fos-like immunoreactivity in lamina terminalis and magnocellular hypothalamic nuclei induced by hypernatremia.
Rats were given either slow (1 h) or rapid (10 min) intravenous infusions of either 6 or 12 mmol NaCl/kg body weight. Fos-like immunoreactivity (FLI) induced by the infusions was measured in several brain regions. The higher dose of NaCl induced FLI in structures of the lamina terminalis, including organum vasculosum (OVLT) and subfornical organ (SFO), as well as in the magnocellular supraoptic (SON) and paraventricular hypothalamic (PVN) nuclei. The lower dose of hypertonic NaCl induced FLI in only the SON and PVN. Faster delivery of the solute load tended to amplify the FLI in SFO and OVLT. These data confirm and extend previous reports of osmotically-induced FLI in rat brain and demonstrate that the discrepancies between these studies result from different dosage regimens of NaCl. The data are discussed as they relate to the lamina terminalis as a primary osmosensitive region in brain.
Animals↗
High prevalence of adeno-associated virus (AAV) type 2 rep DNA in cervical materials: AAV may be sexually transmitted.
Adeno-associated virus (AAV) is a human parvovirus that in laboratory and animal models has the ability to suppress the oncogenic phenotype of a variety of viruses and cellular derived oncogenes. The inhibitory effects of AAV have been mapped to its rap gene (Rep78 protein). Furthermore, seroepidemiologic data indicate that AAV infection is linked to reduced cervical cancer rates in humans. Because of AAV's inverse relationship with cervical cancer, we attempted to identify AAV rep sequences within DNA derived from cervical brushings taken from nondiseased middle class patients at a Little Rock clinic. Polymerase chain reaction (PCR) amplification was carried out with primers designed to amplify a specific segment of the endogenous human beta-globin gene or the AAV rep gene. Of those cervical samples that were positive for beta-globin DNA, 50% were also found to be positive for AAV rep DNA when analyzed by either ethidium bromide staining or dot-blot hybridization with an internal probe. These data strongly suggest that AAV is commonly carried in the genital region and further raise the possibility that AAV can be sexually transmitted.
Cervix Uteri↗
Race and gender differences in the distribution of home and community-based services in Florida.
This article examines the distribution of home and community-based services (HCBS) under Florida's Medicaid waiver program. Controlling for personal and community characteristics, it was found that gender and race significantly affect the access of the disabled adult population to HCBS services, with women and nonwhites significantly more likely to be receiving HCBS services. At the county level, the likelihood of one's being in the waiver program is contingent on the racial composition and level of segregation of the county. People residing in counties with substantial proportions of nonwhites are less likely to receive HCBS services--whatever their race. However, the higher the rate of racial segregation in the county, the higher the probability that the Medicaid disabled adult population will receive HCBS services. The Medicaid waiver program allows older, disabled black women to remain in their home neighborhoods rather than having to move to predominantly white areas where nursing homes are concentrated. Thus, the HCBS program not only provides them with a form of care that is preferred by most older people but also resolves market problems stemming from the lack of nursing homes in segregated areas by taking advantage of support systems in black households.
Aged↗
Elimination of false negative results in the two-hybrid system in the phagocyte NADPH oxidase.
The yeast two-hybrid system is finding increased use in the study of interactions between proteins. In this method, two polypeptides are expressed in yeast as fusion proteins to a transcriptional activator DNA-binding domain (bd) and activating domain (ad), respectively. Interaction between the two polypeptides reconstitutes function of a transactivator which controls expression of reporters. The phagocyte NADPH oxidase is a complex of membrane cytochrome b558 (comprised of subunits p22-phox and gp91-phox) and three cytosol proteins (p47-phox, p67-phox, and p21rac) that translocate to membrane and bind to cytochrome b558. This is the first report to demonstrate that two of cytosolic components of cytochrome b558, p47-phox binding to p67-phox each other. We encountered several methodological problems in the two-hybrid system which are the focus of this report.
False Negative Reactions↗
[Vaginal bleeding patterns and the regularity in use of Norplant].
OBJECTIVES: To understand vaginal bleeding patterns and the regularity in use of Norplant, and find out the main cause of termination that Norplant users can not bear. METHODS: A total of 306 menstrual diaries of Norplant users for 5 years were analyzed. The analysis of bleeding patterns was carried out by using the reference period approach and followed the guidelings published by WHO. RESULTS: The total number of vaginal bleeding days and the number of spotting days in the first reference periods were 36.6 days and 21.5 days, respectively. They were decreased obviously in the third reference period. The number of bleeding days was not obviously changed. The percentage of irregular bleeding was predominent in all of the bleeding patterns, 43.1%-57.6% usually. The percentage of prolonged bleeding was 13.4%-31.0%, secondly. The percentage of "normal" was only 33%. The number of bleeding days and the percentage of prolonged bleeding were more in bleeding termination group than in continuation group. The percentage of irregular bleeding was lower. CONCLUSIONS: During the initial stage of use of Norplant the total number of vaginal bleeding days was increased, but decreased obviously after 6 months. This change was influenced by the number of spotting days. Irregular bleeding and prolonged bleeding were main types of the disturbance of menstrual cycles after use of Norplant, The main cause of termination was prolonged bleeding.
Adult↗