Long-term follow-up of multilayer amniotic membrane transplantation (MLAMT) for non-traumatic corneal perforations or deep ulcers with descemetocele.

Background: To evaluate the long-term efficacy of multilayer amniotic membrane transplantation for reconstruction of epithelium and stroma in non-traumatic corneal perforations (less than 2 mm) or deep ulcers with descemetocele.

Design: Retrospective, non-comparative, interventional case series.

Patients And Methods: Eleven consecutive patients with non-traumatic corneal perforations or deep corneal ulcers with descemetocele refractory to conventional treatments: herpetic or zoster keratitis (n = 4), Sjögren's syndrome (n = 2), rosacea (n = 1), hydrops (n = 1), mucous membrane pemphigoid (n = 1), bacterial keratitis (n = 1) and perforation after protontherapy for melanoma (n = 1).

Glaucoma and keratoprosthesis surgery: role of adjunctive cyclophotocoagulation.

Purpose: To evaluate the efficacy and safety of diode laser transscleral cyclophotocoagulation (DLTSC) to control intraocular pressure (IOP) in keratoprosthesis patients with uncontrolled glaucoma.

Patients And Methods: Between 1993 and 2007, 18 eyes of 18 patients underwent DLTSC, either before (n=3), during (n=1), or after (n=14) keratoprosthesis surgery. Keratoprosthesis type I was used in 72%.

Ten years follow-up after deep sclerectomy with collagen implant.

Purpose: To evaluate the long-term success rate and complications of nonpenetrating deep sclerectomy with collagen implant in open-angle glaucoma.

Patients And Methods: Clinical, prospective, monocentric, nonrandomized, unmasked study on 105 patients with medically uncontrolled glaucoma. A standard procedure deep sclerectomy with collagen implant was performed.

The DNA end-binding protein Ku regulates silencing at the internal HML and HMR loci in Saccharomyces cerevisiae.

Heterochromatin resides near yeast telomeres and at the cryptic mating-type loci, HML and HMR, where it silences transcription of the alpha- and a-mating-type genes, respectively. Ku is a conserved DNA end-binding protein that binds telomeres and regulates silencing in yeast. The role of Ku in silencing is thought to be limited to telomeric silencing.

Ex-PRESS R-50 miniature glaucoma implant insertion under the conjunctiva combined with cataract extraction.

Purpose: To evaluate the efficacy and safety of the Ex-PRESS R-50 implant (Optonol Ltd.) in eyes operated on for open-angle glaucoma combined with phacoemulsification.

Setting: Glaucoma Unit, Ophthalmology Department, University of Lausanne, Lausanne, Switzerland.

Histone H1 regulates chromatin condensation in Leishmania parasites.

We investigated the functional role of the Leishmania histone H1 and demonstrate for the first time that addition of histone H1 has a strong effect on microccocal digestion, chromatin condensation of parasite nuclei and that its overexpression can modulate parasite infectivity in vivo.

Synthetic triacylated lipid A derivative activates antigen presenting cells via the TLR4 pathway and promotes antigen-specific responses in vivo.

Triggering the maturation of dendritic cells (DC) with toll-like receptor (TLR) agonists is a favored strategy for the development of vaccine adjuvants. The triacyl pseudo-dipeptidic agent OM-197-MP-AC mimicking the lipid A structure of endotoxin induces the maturation of human monocyte-derived DC. In this study we investigated the signaling pathway by which this molecule activates DC.

Vaccination against Leishmania major in a CBA mouse model of infection: role of adjuvants and mechanism of protection.

Gp63 is a major surface protein of Leishmania promastigotes. Its protective efficacy has been tested in several experimental models using different mouse strains, gp63 forms, adjuvants and routes of immunization, giving rise to conflicting results. This investigation was designed to determine whether these discrepancies could be ascribed to differing experimental procedures, and to compare gp63-induced protection with that achieved using live promastigotes.

SAS4 and SAS5 are locus-specific regulators of silencing in Saccharomyces cerevisiae.

Sir2p, Sir3p, Sir4p, and the core histones form a repressive chromatin structure that silences transcription in the regions near telomeres and at the HML and HMR cryptic mating-type loci in Saccharomyces cerevisiae. Null alleles of SAS4 and SAS5 suppress silencing defects at HMR; therefore, SAS4 and SAS5 are negative regulators of silencing at HMR. This study revealed that SAS4 and SAS5 contribute to silencing at HML and the telomeres, indicating that SAS4 and SAS5 are positive regulators of silencing at these loci.

Identification of SAS4 and SAS5, two genes that regulate silencing in Saccharomyces cerevisiae.

In Saccharomyces cerevisiae, chromatin-mediated silencing inactivates transcription of the genes at the HML and HMR cryptic mating-type loci and genes near telomeres. Mutations in the Rap1p and Abf1p binding sites of the HMR-E silencer (HMRa-e**) result in a loss of silencing at HMR. We characterized a collection of 15 mutations that restore the alpha-mating phenotype to MATalpha HMRa-e** strains.

Designer deletion and prototrophic strains derived from Saccharomyces cerevisiae strain W303-1a.

We report the construction of Saccharomyces cerevisiae strains isogenic to W303-1a that are designed to allow efficient genetic analysis. To facilitate the generation of null alleles of target genes by PCR-mediated gene disruption, we constructed designer deletion alleles of the ARG4, TRP1 and URA3 genes. In addition, a single pair of oligonucleotide primers were designed that can be used to amplify any of several marker genes for use in PCR-mediated gene disruption.

Identification of a compound origin of replication at the HMR-E locus in Saccharomyces cerevisiae.

Eukaryotic chromosomal origins of replication are best defined in Saccharomyces cerevisiae. Previous analysis of yeast origins suggests that they are relatively simple structures comprised of three or four small DNA sequence elements contained within approximately 100-200-base pair regions (Gilbert, D. M.

HMR-I is an origin of replication and a silencer in Saccharomyces cerevisiae.

There appear to be fundamental differences between the properties of the silencers at HML and HMR, with some being origins of replication and others not. Moreover, past studies have suggested that HMR-I's role in silencing may be restricted to plasmid contexts. This study established that HMR-I, like HMR-E and unlike either HML silencer, is an origin of replication.

Leishmania mexicana: extracellular proton concentration is a key regulator of cysteine proteinase CPb expression.

The purpose of this work was to determine which parameters trigger expression of proteins that are potentially important for the differentiation of Leishmania mexicana from the promastigote to the amastigote stage. To this effect, a protein-free axenic incubation system was used that supported the differentiation of L. mexicana promastigotes into amastigotes at 33 degreesC and at acidic pH.

The role of Sas2, an acetyltransferase homologue of Saccharomyces cerevisiae, in silencing and ORC function.

Silencing at the cryptic mating-type loci HML and HMR of Saccharomyces cerevisiae requires regulatory sites called silencers. Mutations in the Rap1 and Abf1 binding sites of the HMR-E silencer (HMRa-e**) cause the silencer to be nonfunctional, and hence, cause derepression of HMR. Here, we have isolated and characterized mutations in SAS2 as second-site suppressors of the silencing defect of HMRa-e**.

Vaccine development against cutaneous leishmaniasis. Subcutaneous administration of radioattenuated parasites protects CBA mice against virulent Leishmania major challenge.

Experiments described in this paper were aimed at determining whether subcutaneous inoculation of live, avirulent Leishmania major would protect mice against infection by the virulent parasite. To this effect, promastigotes or amastigotes of a highly virulent strain of L. major (MRHO/IR/76), used in human trials of leishmanization, and which induces non-healing skin lesions in both CBA and BALB/c mice, were rendered non-pathogenic by gamma irradiation.

An origin of DNA replication and a transcription silencer require a common element.

A eukaryotic chromosomal origin of replication was identified in the yeast Saccharomyces cerevisiae. By several criteria, including map position, deletion analysis, and a synthetic form of saturation mutagenesis, the origin co-localized with the HMR-E silencer, which is a DNA element that represses transcription of the adjacent genes. A specific site within the silencer was required for both initiation of chromosomal replication and for repression of transcription.

Silencing: the establishment and inheritance of stable, repressed transcription states.

Silencing refers to a particular type of transcriptional repression characterized by the formation of a genetically heritable, repressed transcriptional state. Examples of silencing include position-effect variegation, X-chromosome inactivation, and the repression of the silent mating-type gene loci in yeast. Recent discoveries suggest that silencing in yeast, like silencing in larger eukaryotes, results from a particular chromatin structure that defines a chromosomal domain.

Sequences far downstream from the classical tRNA promoter elements bind RNA polymerase III transcription factors.

We have examined the interaction of transcription factors TFIIIC and TFIIID with a silkworm alanine tRNA gene. Previous functional analysis showed that the promoter for this gene is unusually large compared with the classical tRNA promoter elements (the A and B boxes) and includes sequences downstream from the transcription termination site. The goal of the experiments reported here was to determine which sequences within the full promoter make stable contacts with transcription factors.

The properties of a new polymerase III transcription factor reveal that transcription complexes can assemble by more than one pathway.

We have resolved a previously unidentified factor (TFIIID) that is required for in vitro transcription of polymerase III templates. Our ability to resolve factor D from each of the other components of the transcription machinery (polymerase and transcription factors IIIB and IIIC) allowed us to test the capacity of these separated components to form stable complexes with tRNA genes. We find that none of the individual components binds detectably to tRNA genes, but that certain combinations of transcription factors do bind.

In vivo isotype regulation of humoral responses to dextran B1355 in BALB/c mice. Double-class IgG3/IgM producers as a function of age.

This report shows that, in 8- to 10-month-old BALB/c mice immunized intraperitoneally with dextran B1355, approximately 75% of IgG3 anti-alpha (1----3) polyglucan (anti-dex) plaque-forming cells (PFC) detected in the spleen were identified as double-Ig class producers secreting simultaneously IgG3 and IgM antibodies with the same specificity for the dex epitope. Under the same conditions of immunization, however, IgA anti-dex PFC were mostly single-class secretors. IgA PFC developed in the spleen in highest numbers (equal to IgM), but in Peyer's patches IgA PFC were sevenfold more numerous than IgM.

Age-dependence of the IgA anti-alpha (1 leads to 3) dextran B1355 response in vitro.

The magnitude of the Balb/c mouse IgA anti-alpha (1 leads to 3) dextran B1355 (anti-dex) response in vivo was recently found to be markedly T-cell-dependent and age-dependent. This report demonstrates that the in vitro IgA anti-dex response by mesenteric lymph nodes (MLN) is highly age-dependent and that there is an age-dependent increase of the background IgA anti-dex plaque-forming cell (PFC) response occurring in the absence of added antigen which correlates significantly with the magnitude of the antigen-stimulated response. In aging mice both background and antigen-stimulated IgA anti-dex responses appeared to be significantly higher in MLN than in spleen cultures.

Synergism between iron chelators and complement for bactericidal activity.

Iron-binding agents such as the plasma protein transferrin or the siderophore desferal from Streptomyces pilosus inhibit the growth of pathogenic bacteria, supposedly by interfering with iron uptake by those bacteria. This study shows that anti-Escherichia coli activity exerted by desferal and transferrin can be increased in a synergistic way by complement and anti-E. coli antibodies of normal serum.