Lupus eritematoso sistémico (LES): perfil de severidad al ingreso hospitalario de 44 pacientes sin seguimiento postdiagnóstico / Systemic Lupus Erythematosus (SLE): Severity profile at hospital admisión of 44 patients without post-diagnosis monitoring

El grado que un paciente cumpla con el seguimiento medico y se adhiera a las indicaciones terapéuticas, se ha convertido en un punto esencial en la repercusión que tendrá sobre la evolución de su enfermedad. Es un tema de gran trascendencia sobre todo en enfermedades crónicas. El incumplimiento es grave porque hace ineficaz el tratamiento prescripto, provocando un aumento de la morbilidad. El paciente presentará complicaciones y secuelas que pueden acarrear lesiones de tipo irreversibles. En el Lupus Eritematoso Sistémico, la adherencia al tratamiento es de suma trascendencia en el control de su enfermedad. El presente estudio tiene como objetivo valorar el impacto que provoca la falta de cumplimiento al tratamiento y el abandono al seguimiento médico. Concomitantemente se evaluó la prevalencia del daño en los distintos órganos y su actividad. Los datos obtenidos en el relevamiento de ingreso a nuestro hospital muestran que 44 pacientes con diagnóstico previo de LES abandonaron el seguimiento en su hospital de origen. Se revisaron las historias clínicas de los ingresados entre mayo de 1984 y de junio de 1999. Desde julio de 1999 a mayo de 2005 se obtuvieron datos en forma prospectiva según grilla prediseñada. Se definieron términos según criterios de clasificación, SLEDAI e Indice de Daño (ACR). Se evaluaron las distintas prevalencias, observándose que la gran mayoría de los pacientes presentó actividad multisistémica con mayor prevalencia en riñón. Más del 50% de los casos presentó daño irreversible con alta prevalencia en riñón y sistema nervioso. Los pacientes dañados evidenciaron mayor tiempo de evolución, pero sin diferencias estadísticamente significativas. No encontramos relación entre daño y mantenimiento del tratamiento corticoideo.

The genetics of complex diseases.

Genetic factors influence virtually every human disorder, determining disease susceptibility or resistance and interactions with environmental factors. Our recent successes in the genetic mapping and identification of the molecular basis of mendelian traits have been remarkable. Now, attention is rapidly shifting to more-complex, and more-prevalent, genetic disorders and traits that involve multiple genes and environmental effects, such as cardiovascular disease, diabetes, rheumatoid arthritis and schizophrenia. Rather than being due to specific and relatively rare mutations, complex diseases and traits result principally from genetic variation that is relatively common in the general population. Unfortunately, despite extensive efforts by many groups, only a few genetic regions and genes involved in complex diseases have been identified. Completion of the human genome sequence will be a seminal accomplishment, but it will not provide an immediate solution to the genetics of complex traits.

RPD3 (REC3) mutations affect mitotic recombination in Saccharomyces cerevisiae.

Prior research identified the recessive rec3-1ts mutation in Saccharomyces cerevisiae which, in homozygous diploid cells, confers a conditional phenotype resulting in reduced levels of spontaneous mitotic recombination and loss of sporulation at the restrictive temperature of 36 degrees C. We found that a 3.4-kb genomic fragment that complements the rec3-1ts/rec3-1ts mutation and which maps to chromosome XIV, is identical to RPD3, a gene encoding a histone de-acetylase. Sporulation is reduced in homozygous diploid strains containing the rec3-1ts allele at 24 degrees C, suggesting that this allele of RPD3 encodes a gene product with a reduced function. Sporulation is abolished in diploid strains homozygous for the rpd3Delta or rec3-1ts alleles, as well as in rpd3Delta/rec3-1ts heteroallelic diploids, at the non-permissive temperature. Acid-phosphatase expression has been shown to be RPD3 dependent. We found that acid-phosphatase activity is greater in diploid strains homozygous for the temperature-sensitive rec3-1ts allele than in RPD3/RPD3 strains and increased further when mutant strains are grown at 36 degrees C. We also tested the rpd3Delta/rpd3Delta strains for their effects on spontaneous mitotic recombination. By assaying a variety of intra- and inter-genic recombination events distributed over three chromosomes, we found that in the majority of cases spontaneous mitotic recombination was reduced in diploid rpd3Delta/rpd3Delta cells (relative to a RPD3/RPD3 control). Finally, although 90% of mitotic recombinant events are initiated in the G1 phase of the growth cycle (i.e., before DNA synthesis) we show that RPD3 is not regulated in a cell-cycle-dependent manner. These data suggest that mitotic recombination, in addition to gene expression, is affected by changes in chromatin architecture mediated by RPD3.

Purpura trombocitopanica trombotica (PTT) asociada a poliartritis erosiva seronegativa / Thrombotic thrombocytopenic purpura associated to seronegative arthritis

Paciente, sexo femenino, edad 22 años. Padecio una poliartritis erosiva simetrica, no nodular, seronegativa, F.A.N. negativo, que requirio varias internaciones en el lapso de 2 años por marcado compromiso del estado general que nunca alcanzo a cumplir criterios suficientes para LES ni otras enfermedades del tejido conectivo. Su ultima internacion respondio a la instalacion de un sindrome neurologico de tipo encefalitico, purpura palpable, anemia microangiopatica y trombocitoipenia severa compatible con PTT. Evaluados los diagnosticos diferenciales se la trato con pulsos de esteroides, falleciendo antes de iniciarse la plasmaafaresis. Los hallazgos anatomopatologicos confirmaron el diagnostico de PTT

Purpura trombocitopanica trombotica (PTT) asociada a poliartritis erosiva seronegativa / Thrombotic thrombocytopenic purpura associated to seronegative arthritis

Paciente, sexo femenino, edad 22 años. Padecio una poliartritis erosiva simetrica, no nodular, seronegativa, F.A.N. negativo, que requirio varias internaciones en el lapso de 2 años por marcado compromiso del estado general que nunca alcanzo a cumplir criterios suficientes para LES ni otras enfermedades del tejido conectivo. Su ultima internacion respondio a la instalacion de un sindrome neurologico de tipo encefalitico, purpura palpable, anemia microangiopatica y trombocitoipenia severa compatible con PTT. Evaluados los diagnosticos diferenciales se la trato con pulsos de esteroides, falleciendo antes de iniciarse la plasmaafaresis. Los hallazgos anatomopatologicos confirmaron el diagnostico de PTT

The genomic instability of yeast cdc6-1/cdc6-1 mutants involves chromosome structure and recombination.

When diploid cells of Saccharomyces cerevisiae homozygous for the temperature-sensitive cell division cycle mutation cdc6-1 are grown at a semipermissive temperature they exhibit elevated genomic instability, as indicated by enhanced mitotic gene conversion, mitotic intergenic recombination, chromosomal loss, chromosomal gain, and chromosomal rearrangements. Employing quantitative Southern analysis of chromosomes separated by transverse alternating field gel electrophoresis (TAFE), we have demonstrated that 2N-1 cells monosomic for chromosome VII, owing to the cdc6-1 defect, show slow growth and subsequently yield 2N variants that grow at a normal rate in association with restitution of disomy for chromosome VII. Analysis of TAFE gels also demonstrates that cdc6-1/cdc6-1 diploids give rise to aberrant chromosomes of novel lengths. We propose an explanation for the genomic instability induced by the cdc6-1 mutation, which suggests that hyper-recombination, chromosomal loss, chromosomal gain and chromosomal rearrangements reflect aberrant mitotic division by cdc6-1/cdc6-1 cells containing chromosomes that have not replicated fully.

Nonrandomly-associated forward mutation and mitotic recombination yield yeast diploids homozygous for recessive mutations.

We have employed the analysis of spontaneous forward mutations that confer the ability to utilize L-alpha-aminoadipate as a nitrogen source (alpha-Aa+) to discern the events that contribute to mitotic segregation of spontaneous recessive mutations by diploid cells. alpha-Aa- diploid cells yield alpha-Aa+ mutants at a rate of 7.8 +/- 3.6 x 10(-9). As in haploid strains, approximately 97% (30/31) of alpha-Aa+ mutants are spontaneous lys2-x recessive mutations. alpha-Aa+ mutants of diploid cells reflect mostly the fate of LYS2/lys2-x heterozygotes that arise by mutation within LYS2/LYS2 populations at a rate of 1.2 +/- 0.4 x 10(-6). Mitotic recombination occurs in nonrandom association with forward mutation of LYS2 at a rate of 1.3 +/- 0.6 x 10(-3). This mitotic recombination rate is tenfold higher than that of a control LYS2/lys2-1 diploid. Mitotic segregation within LYS2/lys2-x subpopulations yields primarily lys2-x/lys2-x diploids and a minority of lys2-x aneuploids. Fifteen percent of lys2-x/lys2-x diploids appear to have arisen by gene conversion of LYS2 to lys2-x; 85% of lys2-x/lys2-x diploids appear to have arisen by mitotic recombination in the CENII-LYS2 interval. lys2-1/lys2-1 mitotic segregants of a control LYS2/lys2-1 diploid consist similarity of 18% of lys2-1/lys2-1 diploids that appear to have arisen by gene conversion of LYS2 to lys2-1 and 82% of lys2-1/lys2-1 diploids that appear to have arisen by mitotic recombination in the CENII-LYS2 interval.(ABSTRACT TRUNCATED AT 250 WORDS)

Diploid yeast cells yield homozygous spontaneous mutations.

A leucine-requiring hybrid of Saccharomyces cerevisiae, homoallelic at the LEU1 locus (leu1-12/leu1-12) and heterozygous for three chromosome-VII genetic markers distal to the LEU1 locus, was employed to inquire: (1) whether spontaneous gene mutation and mitotic segregation of heterozygous markers occur in positive nonrandom association and (2) whether homozygous LEU1/LEU1 mutant diploids are generated. The results demonstrate that gene mutation of leu1-12 to LEU1 and mitotic segregation of heterozygous chromosome-VII markers occur in strong positive nonrandom association, suggesting that the stimulatory DNA lesion is both mutagenic and recombinogenic. In addition, genetic analysis of diploid Leu+ revertants revealed that approximately 3% of mutations of leu1-12 to LEU1 result in LEU1/LEU1 homozygotes. Red-white sectored Leu+ colonies exhibit genotypes that implicate post-replicational chromatid breakage and exchange near the site of leu1-12 reversion, chromosome loss, and subsequent restitution of diploidy, in the sequence of events leading to mutational homozygosis. By analogy, diploid cell populations can yield variants homozygous for novel recessive gene mutations at biologically significant rates. Mutational homozygosis may be relevant to both carcinogenesis and the evolution of asexual diploid organisms.

Conditional hyporecombination mutants of three REC genes of Saccharomyces cerevisiae.

We have isolated and characterized three conditional hyporecombination mutants, rec1-1, rec3-1 and rec4-1, that define three REC genes of Saccharomyces cerevisiae required for spontaneous general mitotic interchromosomal recombination. Each MATa/MAT alpha rec/rec diploid is deficient in mitotic single site gene conversion, intragenic recombination, intergenic recombination and sporulation at the restrictive temperature (36 degrees C). The rec1-1 mutation also confers conditional enhanced sensitivity to the killing effects of X-rays. The rec1-1 and rec3-1 mutations have been mapped to chromosome VII. The rec1-1, rec3-1 and rec4-1 mutations exhibit complementation at 36 degrees C for both mitotic recombination and sporulation.

The REC46 gene of Saccharomyces cerevisiae controls mitotic chromosomal stability, recombination and sporulation: cell-type and life cycle stage-specific expression of the rec46-1 mutation.

The recessive hyperrecombination mutation rec46-1, isolated by ultraviolet light mutagenesis of the MAT alpha n+1 chromosome VII disomic strain LBL1 (Esposito et al. 1982), enhances the mitotic rates of spontaneous gene conversion, intergenic recombination and restitution of haploidy (due to chromosomal loss or mitotic nondisjunction) in MAT alpha n+1 chromosome VII disomic strains. The rec46-1 mutation does not prevent HO directed homothallic interconversion of mating types. MATa/MaT alpha rec46-1/rec46-1 diploids exhibit the same degree of hyperrecombinational activity as MAT alpha rec46-1 n+1 chromosome VII disomics with respect to gene conversion and intragenic recombination resulting in prototrophy. When compared to MAT alpha rec46-1 n+1 disomics however, MATa/MAT alpha rec46-1/rec46-1 diploids exhibit a ten fold reduced level of hyperrecombinational activity with respect to intergenic recombination and present no evidence of chromosomal loss or nondisjunction resulting in 2n-1 monosomic segregants. MATa/MAT alpha rec46-1/rec46-1 diploids are sporulation-deficient. The results obtained demonstrate that the REC46 gene product modulates mitotic chromosomal stability and recombination and is essential for sporulation (meiosis and ascospore formation).

Coincident gene conversion during mitosis in saccharomyces.

During mitosis, gene conversion events at the TRP5 locus on chromosome VII are coupled with conversion events at LEU1, a locus 18 cM away, 1200 times more frequently than would be expected for two independent acts of recombination. Such coincident conversion events that occur over relatively long distances could be due to several mechanisms. We discuss these possibilities and describe an experiment that indicates that a portion of coincident events is due to extensive heteroduplexes. The phenomenon of coincident gene conversion is discussed in relation to our earlier evidence that spontaneous recombination between homologues occurs prereplicationally in mitosis.

Molecular mechanisms of recombination in Saccharomyces cerevisiae: testing mitotic and meiotic models by analysis of hypo-rec and hyper-rec mutations.

Recombination in the yeast Saccharomyces cerevisiae has been the subject of extensive genetic studies documenting the general properties of intragenic and intergenic recombination and the differences between mitotic and meiotic gene conversion and reciprocal exchange. Spontaneous mitotic and meiotic events differ in the time of onset of recombination relative to chromosomal replication, symmetry versus asymmetry of putative heteroduplex DNA regions, polarity of conversion of intragenic markers, and the lengths of DNA segments that undergo coincident conversion. The differences observed and the properties of yeast rec mutations provide evidence for multiple modes or pathways of mitotic and meiotic recombination. Several molecular models of recombination have been proposed to account for the basic parameters of genetic recombination and the differences between mitotic and meiotic recombination. Since the models differ with respect to the partial reactions comprising recombination they predict the isolation of different classes of hypo-recombination and hyper-recombination rec mutants. We have isolated a broad spectrum of yeast REC gene mutations that includes both hyper-rec and hypo-rec mutants. Five phenotypic classes of rec variants have been identified based upon their effects on spontaneous mitotic gene conversion and intergenic recombination. Their characteristics demonstrate that mitotic gene conversion and intergenic recombination are under independent as well as coordinate genetic control. Four gene mutations affecting recombination rad50, rad52, rem1 and spo11 have been extensively examined in several laboratories and illustrate the information that can be obtained by characterization of double mutant strains, detailed genotypic analysis of recombinants, and studies of meiotic recombination in cells in which the reductional division of meiosis has been bypassed by the spo13 mutation.

Enhancement of spontaneous mitotic recombination by the meiotic mutant spo11-1 in Saccharomyces cerevisiae.

Both nonreciprocal and reciprocal mitotic recombination are enhanced by the recessive mutant spo11-1, which was previously shown to affect meiosis by decreasing recombination and increasing nondisjunction. The mitotic effects are not distributed equally in all chromosomal regions. The genotypes of mitotic recombinants in spo11-1/spo11-1 diploid cells provide further evidence that widely spaced chromosomal markers undergo coincident conversion in mitosis.

Simultaneous detection of changes in chromosome number, gene conversion and intergenic recombination during mitosis of Saccharomyces cerevisiae: spontaneous and ultraviolet light induced events.

We have employed a hyperhaploid strain of Saccharomyces cerevisiae disomic for chromosome VII to monitor spontaneous and ultraviolet light induced restitution of haploidy (chromosomal loss and/or nondisjunction), mitotic gene conversion and mitotic intergenic recombination. The disomic chromosomal pair incorporates six heterozygous markers, including cyh2 (r), distributed on both sides of the centromere. Cycloheximide resistant segregants of spontaneous origin were analyzed to calculate the spontaneous mitotic rates of restitution of haploidy, intergenic recombination and gene conversion that result in expression of the cyh2 (r) mutation. Restitution of haploidy was found to be the most common source of spontaneously arising cycloheximide resistant segregants. In contrast, those induced by ultraviolet light resulted most frequently from gene conversion of CYH2 (s) to cyh2 (r). The chromosome VII hyperhaploid system provides a sensitive method to detect the aneugenic and recombinagenic effects of suspect chemical and physical agents.

Mitotic recombination: mismatch correction and replicational resolution of Holliday structures formed at the two strand stage in Saccharomyces.

In a preliminary report (Esposito 1978), evidence was presented which showed that heteroallelic recombination resulting in prototrophic colonies occurs at the 2-strand stage. A model utilizing replicative resolution of Holliday structures was proposed to explain how gene conversion at the 2-strand stage can result in exchange of outside markers. The object of the experiments reported herein was to present detailed genetic evidence for 2-strand recombination. In addition, we examined the features of mitotic recombination with respect to symmetry, length and polarity of heteroduplexes in wild type strains (REM1/REM1) and in strains bearing the hyper-recombination mutation rem1-1. To do this, we constructed strains so that prototrophs arising from heteroallelic recombination and recombinant for outside markers were detected by visual inspection. By analyzing these colonies genetically, we have inferred several features of mitotic recombination which distinguish it from its meiotic counterpart. Firstly, mitotic heteroduplexes are often symmetric while meiotic heteroduplexes are almost exclusively asymmetric. Secondly, heteroduplexes tend to be longer in mitosis that in meiosis. Thirdly, unlike meiotic conversion, mitotic conversion does not show strong polarity. Recombination in strains homozygous for the rem1-1 mutation also takes place at the 2-strand stage. The rem1-1 mutation, however, appears to alter the features of mismatch correction.

Erythromycin and cycloheximide sensitivities of protein and RNA Synthesis in sporulating cells of Saccharomyces cerevisiae: Environmentally induced modifications controlled by chromosomal and mitochondrial genes.

The purpose of the experiments reported below was to examine the response in sporulation medium of the three diploid cell types MATα MATα, MATα MATα (asporogenic diploids) and MATα MATα (sporogenic diploid) to erythromycin, a specific inhibitor of mitochondrial protein synthesis (MPS) in vegetative cultures, and cycloheximide, a specific inhibitor of cytosol protein synthesis (CPS) in vegetative cultures. When MATα MATα diploids are transferred to sporulation medium a significant fraction of total protein synthesis (CPS + MPS) becomes sensitive to erythromycin in contrast to the behavior of MATa MATa and MATα MATα diploids in which the resistance of CPS to erythromycin is maintained. The decompartmentalization of erythromycin sensitivity is thus cell type specific. Erythromycin stimulates total RNA synthesis of MATα MATα cells in sporulation medium but not of MATα MATα and MATα MATα cells. Cycloheximide inhibits protein synthesis and stimulates RNA synthesis in all three diploid cell types. An erythromycin resistant mutant, shown to be due to a mutation of the mitochondrial genome, exhibited only partial resistance of CPS to erythromycin in sporulation medium in the background of the MATα MATα mating type genotype. Total RNA synthesis in this mutant was not stimulated. The results reported indicate that mitochondrial functions during sporulation are not restricted to those involving respiratory metabolism.