The prevalence, incidence, and clinical assessment of neuromyelitis optica spectrum disorder in patients with demyelinating diseases.

BACKGROUND: Neuromyelitis optica spectrum disorder (NMOSD) is characterised by recurrent attacks of optic neuritis and transverse myelitis. The purpose of this work was to identify the incidence and prevalence of NMOSD and its clinical characteristics in the population treated for demyelinating diseases in Western Mexico. MATERIAL AND METHOD: A descriptive, retrospective study was carried out in the Department of Neurology, at the Sub-specialty Medical Unit, Specialties Hospital (known by its Spanish abbreviation UMAE-HE), of the National Western Medical Center (CMNO), Mexican Institute of Social Security (IMSS). A review of the electronic files for all patients with a diagnosis of NMOSD in 2019, was carried out in the State of Jalisco, Mexico. RESULTS: Fifty-eight patients with NMOSD were included in the study. The incidence was 0.71/100 000 (CI 0.60-0.85) and the prevalence was 1.09/100 000 (CI 0.84-1.42). There were 79.3% women, and 20.6% were men (P = .01). All (100%) patients presented with anti-aquaporin-4 immunoglobulin G, and 89.6% showed seropositivity for anti-aquaporin-4 (CI 82.6-94.9). Magnetic resonance imaging was performed on 100% of patients, where 34.4% were normal, and 65.5% (38) abnormal, presenting with non-specific subcortical lesions (P = 0.04). The initial clinical presentation was optic neuritis (ON) in 58.6%; where 31.0% was bilateral ON, 20.7% was left ON, and 6.9% were right ON; transverse myelitis in 26.0%, area postrema syndrome (APS) in 10.3%, among others. CONCLUSIONS: The incidence of NMOSD exceeds 0.71/100 000, the prevalence is low at 1.09/100 000, and NMOSD is predominantly found in women.

FALSIFLORA, the tomato orthologue of FLORICAULA and LEAFY, controls flowering time and floral meristem identity.

Characterization of the tomato falsiflora mutant shows that fa mutation mainly alters the development of the inflorescence resulting in the replacement of flowers by secondary shoots, but also produces a late-flowering phenotype with an increased number of leaves below first and successive inflorescences. This pattern suggests that the FALSIFLORA (FA) locus regulates both floral meristem identity and flowering time in tomato in a similar way to the floral identity genes FLORICAULA (FLO) of Antirrhinum and LEAFY (LFY) of Arabidopsis. To analyse whether the fa phenotype is the result of a mutation in the tomato FLO/LFY gene, we have cloned and analysed the tomato FLO/LFY homologue (TOFL) in both wild-type and fa plants following a candidate gene strategy. The wild-type gene is predicted to encode a protein sharing 90% identity with NFL1 and ALF, the FLO/LFY-like proteins in Nicotiana and Petunia, and about 80 and 70% identity with either FLO or LFY. In the fa mutant, however, the gene showed a 16 bp deletion that results in a frameshift mutation and in a truncated protein. The co-segregation of this deletion with the fa phenotype in a total of 240 F2 plants analysed supports the idea that FA is the tomato orthologue to FLO and LFY. The gene is expressed in both vegetative and floral meristems, in leaf primordia and leaves, and in the four floral organs. The function of this gene in comparison with other FLO/LFY orthologues is analysed in tomato, a plant with a sympodial growth habit and a cymose inflorescence development.

Lisofylline sensitizes p53 mutant human ovarian carcinoma cells to the cytotoxic effects of cis-diamminedichloroplatinum (II).

A novel approach to counteracting drug resistance is the development of nontoxic agents that are able to preferentially increase the sensitivity of tumor cells to the cytotoxicity of chemotherapy. The possibility that such an agent could be directed specifically against p53-defective tumor cells led us to study the new methylxanthine, Lisofylline, for its ability to sensitize ovary cancer cells to cis-diamminedichloroplatinum(II) (CDDP). In cell lines lacking functional p53 (SKOV3, SKOV3 CDDP-resistant, OVCAR3, and OVCAR432) Lisofylline (20-100 microM) enhanced the cytotoxicity of CDDP by approximately 50% as measured by the Alamar blue vital dye indicator assay. LSF had no effect on p53 wild-type cell lines: OVCAR 420, 429, and 433. Restoration of wild-type p53 phenotype by transfection of SKOV3 cells with a p53 cDNA expression vector showed reversal of sensitization by Lisofylline to CDDP cytotoxicity. While sensitization to DNA damaging agents by other methylxanthines is related to an abrogation of G2 delay, FACS data found no loss of CDDP-induced G2 block in the cell lines, demonstrating that Lisofylline enhanced sensitization. Cell death was examined by quantitative fluorescence microscopy but no increase in apoptosis attributable to Lisofylline exposure was observed. Our results show that the combination of CDDP and Lisofylline preferentially sensitizes p53-defective cancer cells to the cytotoxic effect of CDDP by a yet undetermined mechanism.

UCN-01 in ovary cancer cells: effective as a single agent and in combination with cis-diamminedichloroplatinum(II)independent of p53 status.

Our goal was to determine the cytotoxicity of 7-OH-hydroxystaurosporine (UCN-01) as a single agent and in combination with cis-diamminedichloroplatinum(II) (CDDP) in a panel of ovarian carcinoma cells. We sought to examine the role of p53 gene function and alterations in cell cycle progression or other mechanisms of action of UCN-01 including perturbation of the apoptosis pathway mediated by NF-kappaB and Bcl-2/Bax. Cytotoxicity was determined from dose-response curves established by the Alamar blue vital dye indicator assay. Restoration of wild-type p53 in a p53 null cell line, SKOV 3, was achieved by transfection of a p53 expression vector. Cell cycle distribution was measured by fluorescence-activated cell sorting analysis of ethidium bromide-stained nuclei. Apoptosis was measured by quantitative fluorescence microscopy. NF-kappaB DNA binding activity was measured by electrophoretic mobility shift assay. Bcl-2 and Bax levels were determined by Western immunoblotting. UCN-01 was effective as a cytotoxic agent alone and in combination with CDDP in all cell lines studied, regardless of p53 status. The degree of sensitization to CDDP conferred by UCN-01, however, was found to correlate with p53 gene status. p53 wild-type cells seem to be more sensitive to the cytotoxic effects of the combination of UCN-01 + CDDP than the p53 mutant cells. This was confirmed in cells in which p53 wild-type function was restored by transfection of p53 cDNA, but these cells are also significantly more sensitive to CDDP alone. The effects of UCN-01 on cell cycle progression also appear to be p53 dependent but may not be the primary mechanism of action. The rate of apoptosis is increased 4-fold in UCN-01 + CDDP-treated cells compared to either agent alone. UCN-01 does not effect NF-kappaB DNA binding activity or Bcl-2 and Bax levels. UCN-01 enhances CDDP cytotoxicity and apoptosis in ovary cancer cells. This occurs regardless of p53 status, but wild-type p53 seems to increase the degree of sensitization.

Encapsulation of the UV-DNA repair enzyme T4 endonuclease V in liposomes and delivery to human cells.

T4 endonuclease V, a pyrimidine-dimer-specific DNA repair enzyme, was encapsulated in liposomes under mild conditions. The encapsulated enzyme was active, and when applied to ultraviolet (UV)-irradiated human cells in culture, the liposomes increased incision of UV-irradiated cellular DNA, enhanced DNA repair replication, and enhanced survival of UV-irradiated cells. This method is a first step in a new approach for topical application of DNA repair enzymes to human skin to prevent skin cancer.

Polyclonal antibodies against O6-methylguanine-DNA methyltransferase in adapted bacteria.

The similarity of the adaptive response and the methyltransferase component in bacterial strains from different phylogenic groups was investigated. An adaptive response with induction of transferase activity was found for the first time in the soil bacteria P. aeruginosa and X. maltophilia. Polyclonal antibodies against the E. coli ada protein were used to investigate the structural similarity of the transferases from several bacterial strains with adaptive responses and inducible transferase activity. These antibodies cross-reacted with transferase from M. luteus and P. aeruginosa but not with proteins from other related bacteria, and not with human transferase. The phylogenic relationships of bacteria with adaptive responses suggest that the response likely was present in the common ancestor of eubacteria. The restricted antibody cross-reactivity may reflect the dual role of the E. coli ada protein not only in DNA repair but in positive gene regulation.

Cultivos primarios de queratinocitos epidérmicos humanos utilizando una capa alimentadora combinada / Primary cultures of human epidermal keratinocytes using a compound feeder layer

Una de las aplicaciones prácticas más importantes del cultivo de queratinocitos epidérmicos humanos es la obtención de sábanas de epidermis, para utilizarlas en el tratamiento de pacientes que han recibido grandes quemaduras y que con la utilizacuón de los métodos convencionales tienen pocas posibilidades de recuperación. Estos cultivos, comúnmente desarrollados sobre un soporte de células SWISS 3T3 letalmente irradiadas, se hicieron crecer sobre un nuevo soporte combinado, primero de fibroblastos primarios de embrión de ratón y posteriormente en presencia de fibroblastos diploides humanos de la propia muestra de piel. Se logró obtener un epitelio escamoso estratificado, comprobado con la tinción de azul de rodanilo, el cual fue desprendido posteriormente de la placa de cultivo. Ensayando la línea celular NIH 3T3 como capa alimentadora, no se obtuvieron resultados positivos

Cultivos primarios de queratinocitos epidérmicos humanos utilizando una capa alimentadora combinada / Primary cultures of human epidermal keratinocytes using a compound feeder layer

Una de las aplicaciones prácticas más importantes del cultivo de queratinocitos epidérmicos humanos es la obtención de sábanas de epidermis, para utilizarlas en el tratamiento de pacientes que han recibido grandes quemaduras y que con la utilizacuón de los métodos convencionales tienen pocas posibilidades de recuperación. Estos cultivos, comúnmente desarrollados sobre un soporte de células SWISS 3T3 letalmente irradiadas, se hicieron crecer sobre un nuevo soporte combinado, primero de fibroblastos primarios de embrión de ratón y posteriormente en presencia de fibroblastos diploides humanos de la propia muestra de piel. Se logró obtener un epitelio escamoso estratificado, comprobado con la tinción de azul de rodanilo, el cual fue desprendido posteriormente de la placa de cultivo. Ensayando la línea celular NIH 3T3 como capa alimentadora, no se obtuvieron resultados positivos