A conceptual approach for scientifically based guidelines.
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Avitaminosis↗
Biomedical subjects
P Walter
Publications and source records attributed to P Walter.
Ire1p: a kinase and site-specific endoribonuclease.
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Endoribonucleases↗
[Chemistry of cosmetics in antiquity].
Several texts, statues and paintings denote the importance of make up and eye medicines since the earliest periods of Egyptian history. We have investigated cosmetic powders that were preserved in original alabaster and reed containers. Quantitative crystallographic and chemical analysis of the mineral and organic components revealed surprising facts. In addition to the well known galena PbS and cerussite PbCO3, two unexpected constituents have been identified: laurionite PbOHCl and phosgenite Pb2 (CO3) Cl2, which are rare halide minerals found in lead slag only in certain places where the sea water has weathered lead debris left over from silver mining operations in Antiquity. Alteration of natural lead minerals is also unlikely, given the excellent state of conservation of the reed vessels. This evidence indicates that laurionite and phosgenite were synthesised artificially. Support for this statement comes from recipes of medicinal products to be "used in ophthalmology" reported by Greco-Roman authors such as Dioscorides and Pline (1st Century B.C.): silver foam PbO is crushed and mixed with rock salt and sometimes with natron (Na2CO3). The reaction seems to be straightforward. However, our experiments in the laboratory have shown a major difficulty, arising from the concomitant production of alkali, which raises the pH and leads to different products. It follows that the Egyptians very early mastered this kind of chemical synthesis and technology, a fact of great importance in the History of Sciences. Fire-based technology had been mastered to manufacture Egyptian Blue pigments since the third millennium B.C. The present results now suggest that wet chemistry was already known 4000 years ago. This key finding provides a new insight into the chemical technology of far greater antiquity than has previously been believed. Yet, an important question remains relative to the ultimate motivation for these technological developments. If the Egyptians initially only aimed at different nuances of grey, they could use cerussite, an easily accessible mineral, crushed and mixed with galena. Thus, one should seek other motivations; we shall propose to consider a religious-ritual context, ultimately linked to a pharmaceutical aspect.
Cosmetics↗
Pausing to decide.
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Alternative Splicing↗
Prm1p, a pheromone-regulated multispanning membrane protein, facilitates plasma membrane fusion during yeast mating.
Cell fusion occurs throughout development, from fertilization to organogenesis. The molecular mechanisms driving plasma membrane fusion in these processes remain unknown. While yeast mating offers an excellent model system in which to study cell fusion, all genes previously shown to regulate the process act at or before cell wall breakdown; i.e., well before the two plasma membranes have come in contact. Using a new strategy in which genomic data is used to predict which genes may possess a given function, we identified PRM1, a gene that is selectively expressed during mating and that encodes a multispanning transmembrane protein. Prm1p localizes to sites of cell-cell contact where fusion occurs. In matings between Deltaprm1 mutants, a large fraction of cells initiate zygote formation and degrade the cell wall separating mating partners but then fail to fuse. Electron microscopic analysis reveals that the two plasma membranes in these mating pairs are tightly apposed, remaining separated only by a uniform gap of approximately 8 nm. Thus, the phenotype of Deltaprm1 mutants defines a new step in the mating reaction in which membranes are juxtaposed, possibly through a defined adherence junction, yet remain unfused. This phenotype suggests a role for Prm1p in plasma membrane fusion.
Amino Acid Sequence↗
The unfolded protein response regulates multiple aspects of secretory and membrane protein biogenesis and endoplasmic reticulum quality control.
The unfolded protein response (UPR) is an intracellular signaling pathway that relays signals from the lumen of the ER to activate target genes in the nucleus. We devised a genetic screen in the yeast Saccharomyces cerevisiae to isolate mutants that are dependent on activation of the pathway for viability. Using this strategy, we isolated mutants affecting various aspects of ER function, including protein translocation, folding, glycosylation, glycosylphosphatidylinositol modification, and ER-associated protein degradation (ERAD). Extending results gleaned from the genetic studies, we demonstrate that the UPR regulates trafficking of proteins at the translocon to balance the needs of biosynthesis and ERAD. The approach also revealed connections of the UPR to other regulatory pathways. In particular, we identified SON1/RPN4, a recently described transcriptional regulator for genes encoding subunits of the proteasome. Our genetic strategy, therefore, offers a powerful means to provide insight into the physiology of the UPR and to identify novel genes with roles in many aspects of secretory and membrane protein biogenesis.
Cloning, Molecular↗
Role of 4.5S RNA in assembly of the bacterial signal recognition particle with its receptor.
The mechanism by which a signal recognition particle (SRP) and its receptor mediate protein targeting to the endoplasmic reticulum or to the bacterial plasma membrane is evolutionarily conserved. In Escherichia coli, this reaction is mediated by the Ffh/4.5S RNA ribonucleoprotein complex (Ffh/4.5S RNP; the SRP) and the FtsY protein (the SRP receptor). We have quantified the effects of 4.5S RNA on Ffh-FtsY complex formation by monitoring changes in tryptophan fluorescence. Surprisingly, 4.5S RNA facilitates both assembly and disassembly of the Ffh-FtsY complex to a similar extent. These results provide an example of an RNA molecule facilitating protein-protein interactions in a catalytic fashion.
Bacterial Proteins↗
Functional and genomic analyses reveal an essential coordination between the unfolded protein response and ER-associated degradation.
The unfolded protein response (UPR) regulates gene expression in response to stress in the endoplasmic reticulum (ER). We determined the transcriptional scope of the UPR using DNA microarrays. Rather than regulating only ER-resident chaperones and phospholipid biosynthesis, as anticipated from earlier work, the UPR affects multiple ER and secretory pathway functions. Studies of UPR targets engaged in ER-associated protein degradation (ERAD) reveal an intimate coordination between these responses: efficient ERAD requires an intact UPR, and UPR induction increases ERAD capacity. Conversely, loss of ERAD leads to constitutive UPR induction. Finally, simultaneous loss of ERAD and the UPR greatly decreases cell viability. Thus, the UPR and ERAD are dynamic responses required for the coordinated disposal of misfolded proteins even in the absence of acute stress.
Endoplasmic Reticulum, Rough↗
Perspectives: structural biology. SRP--where the RNA and membrane worlds meet.
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Amino Acid Motifs↗
Transplantation of autologous iris pigment epithelium after removal of choroidal neovascular membranes.
BACKGROUND: Transplantation of autologous iris pigment epithelium (IPE) into the subretinal space has been suggested as one approach for the treatment of age-related macular degeneration, as well as for other conditions in which loss of retinal pigment epithelium (RPE) occurs. Surgical removal of choroidal neovascular membranes is associated with traumatic loss of the RPE cell layer, disruption of the integrity of the photoreceptor-RPE complex, and limited visual outcome. OBJECTIVE: To examine whether IPE cells can substitute for RPE cells to be transplanted to the subretinal space of patients with either RPE degenerative disease or traumatic loss of the RPE cell layer after subretinal surgery. METHODS: Autologous IPE cells were transplanted to the subretinal space in 20 consecutive patients undergoing removal of subretinal fibrovascular membranes using pars plana vitrectomy. Autologous IPE cells were harvested by iridectomy, isolated, and transplanted directly to the subretinal spaces. Transplants were evaluated for 6 to 11 months by funduscopy, fluorescein angiography, and scanning laser ophthalmoscopic (SLO) microperimetry. RESULTS: For the entire follow-up period, no evidence of any immunologic response was observed. Revisional surgery was necessary in 3 patients because of complications (rhegmatogenous retinal detachment [n = 1]; proliferative vitreoretinopathy [n = 1]; and macular pucker [n = 1]); 1 patient did not receive IPE cells. Five of 19 phakic eyes underwent cataract surgery; in 1 case this was combined with the vitrectomy. Five patients showed improved visual acuity of 3 to 4 lines, 13 patients had stable visual acuity (+/-2 lines), and 2 patients had reduced visual acuity of 6 lines. CONCLUSIONS: In this pilot study, the transplantation of autologous IPE cells was done as an addition to conventional surgical excision of choroidal neovascular membranes. Transplanted cells were well tolerated in the subretinal space and did not adversely affect the function of the photoreceptors, since improvement or stable visual acuity was observed in 18 patients after IPE transplantation. These results suggest that autologous IPE cells may be used as a substitute for autologous RPE cells to transplant to the subretinal space to treat age-related macular degeneration.
Adult↗
Evoked cortical potentials after electrical stimulation of the inner retina in rabbits.
BACKGROUND: Electrical stimulation of the retina using implantable devices may be one possible approach in the treatment of blindness causing progressive degeneration of the outer retina. Stimulation of ganglion cells or fibers could be achieved by epiretinal positioning of a microelectrode array as one component of a retinal prosthesis. Experiments were performed in rabbits to determine whether it is possible to elicit cortical responses with current pulses delivered via an epiretinal placed microelectrode array. METHODS: Polyimide-based microelectrode arrays with platinum electrodes and silicon-based TiN electrode arrays were implanted and placed onto the retinal surface of healthy pigmented rabbits after vitrectomy. The devices were temporarily fixated under PFCL and connected to constant current sources delivering bi- or monophasic current pulses. Cortical evoked potentials were recorded using subcutaneously implanted EEG electrodes over the visual cortex. RESULTS: The surgical procedures for implantation and fixation could be performed without serious complications. The electrode array could be placed in the area of the visual streak. Cortical potentials could be recorded after pulse train stimulation either with biphasic or with monophasic pulses. At threshold, pulses of 10 microA/phase and 100 micros/phase were necessary for detection of an electrically evoked cortical potential (EEP). Charge delivery at threshold varied between 0.1 and 0.3 nC/phase and charge densities varied between 1 and 12 microC/cm2. The EEP amplitude increased with increasing stimulus strength. The cortical response could be completely blocked with a retrobulbar injection of 4 ml lidocaine 2%. CONCLUSION: Constant current pulses delivered via Retina Stimulator devices equipped either with platinum electrodes or with TiN electrodes placed onto the inner retinal surface were able to elicit cortical potentials in the rabbit visual system. At threshold the charge delivery seems to be in a safe range.
Animals↗
Mandibular pseudocarcinomatous hyperplasia.
AIMS: Three unusual cases of pseudocarcinomatous (pseudoepitheliomatous) hyperplasia (PH) affecting chronic osteomyelitic mandibular sequestra are reported to highlight the differences with the various squamous neoplasms which occur in that site. METHODS AND RESULTS: In two patients carrying a mandibular graft following the excision of an ameloblastoma, mucosal ulcers resulted in chronic osteomyelitis. In a third patient, an apical dental infection was associated with fistulated osteomyelitis. Histology of the three sequestra showed an intraosseous squamous proliferation. It was characterized by a peripheral involvement of medullary spaces, the more mature epithelial layer covering the bone trabeculae without intervening stroma, and the basal type epithelial layer surrounding a central fibrovascular core. There were no histological or cytological signs of malignancy. CONCLUSION: PH shows an inverted pattern when compared with the centro-medullary tumoural islands seen in the various oral or odontogenic squamous neoplasms which occur in the jaws. The lack of signs of malignancy distinguish PH from common squamous cell carcinomas. A short clinical course is an important feature in the distinction of PH from the well differentiated squamous cell carcinomas which may develop in fistulated chronic osteomyelitis.
Adolescent↗
Influence of membrane differential filtration on the natural course of age-related macular degeneration: a randomized trial.
PURPOSE: Membrane differential filtration is able to optimize rheologic parameters by eliminating high molecular weight proteins and lipoproteins from the blood. Following the hypothesis that these changes result in an improvement of the microcirculation, the authors tested the efficacy of membrane differential filtration in improving visual function in patients with age-related macular degeneration (ARMD). METHODS: Forty patients (40 eyes) were randomized into two groups. The treatment group was treated five times over a period of 21 weeks. In both groups, 9/20 of the eyes showed subfoveolar subretinal neovascularization. The main parameter of the study was visual acuity (VA). Electroretinogram (ERG), electrooculogram, and macular visual evoked potentials were also recorded. Plasma and whole blood viscosity and erythrocyte aggregation were measured. RESULTS: The 20 patients treated repeatedly over a period of 21 weeks showed a mean improvement of 0.63 lines (SD 1.8) of VA on Early Treatment Diabetic Retinopathy Study charts. The control group showed a deterioration of 0.94 lines (SD 1.7) compared to VA at baseline examination. The amplitude of the ERG photopic a-wave and the flicker ERG was significantly increased. The rheologic parameters were lowered in all treated patients. CONCLUSION: Repetitive treatment with membrane differential filtration is able to improve visual acuity of patients with ARMD and the natural course of this disease. Several questions arise from the results of this study. Further research will show if it is possible to optimize the selection of patients for subgroups with predictive responses through morphologic and functional tests and how to create an optimized and individual treatment strategy determined by the quality, intensity, and frequency of treatment sessions.
Aged↗
L-Carnitine, a 'Vitamin-like Substance' for functional food. Proceedings Of the symposium on L-carnitine, april 28 to may 1, 2000, zermatt, switzerland
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Journal Article↗
A role for presenilin-1 in nuclear accumulation of Ire1 fragments and induction of the mammalian unfolded protein response.
The unfolded protein response (UPR) mediates signaling from the endoplasmic reticulum to the nucleus. In yeast, a key regulatory step in the UPR is the spliceosome-independent splicing of HAC1 mRNA encoding a UPR-specific transcription factor, which is initiated by the transmembrane kinase/endoribonuclease Ire1. We show that yeast HAC1 mRNA is correctly spliced in mammalian cells upon UPR induction and that mammalian Ire1 can precisely cleave both splice junctions. Surprisingly, UPR induction leads to proteolytic cleavage of Ire1, releasing fragments containing the kinase and nuclease domains that accumulate in the nucleus. Nuclear localization and UPR induction are reduced in presenilin-1 knockout cells. These results suggest that the salient features of the UPR are conserved among eukaryotic cells and that presenilin-1 controls Ire1 proteolysis in mammalian cells.
Animals↗
Blobel's nobel: a vision validated.
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Cell Physiological Phenomena↗
The crystal structure of the dimerization initiation site of genomic HIV-1 RNA reveals an extended duplex with two adenine bulges.
BACKGROUND: An important step in retroviral replication is dimerization of the genomic RNA prior to encapsidation. Dimerization is initiated by the formation of a transient 'kissing-loop complex' that is thought to be subsequently matured into an extended duplex by the nucleocapsid protein (NCp). Although chemical probing and nuclear magnetic resonance spectroscopy have provided insight into the structure of the kissing-loop structure, no structural information concerning the extended-duplex state is available so far. RESULTS: The structure of a minimal HIV-1 RNA dimerization initiation site has been solved at 2.3 A resolution in two different space groups. It reveals a 22 base pair extended duplex with two noncanonical Watson-Crick-like G-A mismatches, each adjacent to a bulged-out adenine. The structure shows significant asymmetry in deep groove width and G-A base-pair conformations. A network of eight magnesium cations was clearly identified, one being unusually chelated by the 3' phosphate of each bulge across an extremely narrowed deep major groove. CONCLUSIONS: These crystal structures represent the putative matured form of the initial kissing-loop complex. They show the ability of this self-complementary RNA hairpin loop to acquire a more stable extended duplex structure. Both bulged adenines form a striking 'base grip' that could be a recognition signal, either in cis for another viral RNA sequence, or in trans for a protein, possibly the NCp. Magnesium binding might be important to promote and stabilize the observed extrahelical conformation of these bulges.
Adenine↗
Mechanism of non-spliceosomal mRNA splicing in the unfolded protein response pathway.
The unfolded protein response is an intracellular signaling pathway that, in response to accumulation of misfolded proteins in the lumen of the endoplasmic reticulum (ER), upregulates transcription of ER resident chaperones. A key step in this pathway is the non-conventional, regulated splicing of the mRNA encoding the positive transcriptional regulator Hac1p. In the yeast Saccharomyces cerevisiae, the bifunctional transmembrane kinase/endoribonuclease Ire1p cleaves HAC1 mRNA at both splice junctions and tRNA ligase joins the two exons together. We have reconstituted HAC1 mRNA splicing in an efficient in vitro reaction and show that, in many ways, the mechanism of HAC1 mRNA splicing resembles that of pre-tRNA splicing. In particular, Ire1p endonucleolytic cleavage leaves 2', 3'-cyclic phosphates, the excised exons remain associated by base pairing, and exon ligation by tRNA ligase follows the same chemical steps as for pre-tRNA splicing. To date, this mechanism of RNA processing is unprecedented for a messenger RNA. In contrast to the striking similarities to tRNA splicing, the structural features of the splice junctions recognized by Ire1p differ from those recognized by tRNA endonuclease. We show that small stem-loop structures predicted to form at both splice junctions of HAC1 mRNA are required and sufficient for Ire1p cleavage.
Base Pairing↗