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Biomedical subjects

L Sperling

Publications and source records attributed to L Sperling.

At least 19 recordsLinked to original sources
PubMed2006

Twin pregnancy possibly associated with high semen quality.

BACKGROUND: Recent studies found an association between a long waiting time to pregnancy (TTP) and reduced probability of twinning and a reduced dizygotic (DZ) twinning rate in subfertile men. However, it remains unsolved whether semen quality is associated with twin offspring. We therefore studied the semen quality in a group of fathers of naturally conceived twins. METHODS: In this study, 37 fathers of DZ twins and 15 fathers of monozygotic (MZ) twins participated, and 349 normal fertile men served as a reference group. All men delivered a semen sample, underwent a physical examination and completed a questionnaire. RESULTS: After adjustment, fathers of DZ and MZ twins had 3.6 (95% CI 1.7; 5.4) and 4.6 (95% CI 2.0; 7.2) percentage points higher percentage of sperm cells with normal morphologic features and percentages of motile sperm cells were 11.5 (95% CI 7.2; 15.9) and 12.5 (95% CI 6.3; 18.6) percentage points higher than the reference group (P < 0.01). Fathers of DZ twins and MZ twins had 24.7 (95% CI; -9.1; 71.3) and 17.0% (-25.2%; 83.0%) higher sperm concentration than the reference group. CONCLUSIONS: Fathers of DZ twins had a better semen quality than the reference group, which supports the assumption that spontaneous DZ twinning rate can be used as a sensor of male fecundity of a population.

Female↗
PubMed2006

Naturally conceived twins with monochorionic placentation have the highest risk of fetal loss.

OBJECTIVE: The aim of this study was to estimate the rate of fetal loss in dichorionic (DC) and monochorionic (MC) twin pregnancies stratified according to zygosity and method of conception. METHODS: In a prospective multicenter observational study women with a twin pregnancy had an ultrasound scan before 14 + 6 weeks' gestation in order to determine chorionicity. The fetal loss rate, the perinatal, neonatal and infant mortality rates and the frequency of very preterm labor were estimated for the different types of twin. RESULTS: Among the 495 pregnancies (421 DC and 74 MC) 229 (46%) were conceived naturally and 266 (54%) by assisted reproduction (AR). Outcome data for 945 liveborn babies were obtained. The spontaneous miscarriage rate before 24 weeks' gestation was 10.9% (7/64) among naturally conceived MC compared to 3.0% (5/165) for naturally conceived DC twins (P < 0.05). For twins conceived by AR the corresponding figures were 0% (0/10) and 0.4% (1/256). The odds ratio (OR) for very preterm birth-before 28 weeks' gestation-was 4.2 for MC twins compared to DC twins. The relative risk of fetal loss or death among DC twins was 20% of the risk for MC twins. CONCLUSION: The risk of fetal loss, very preterm delivery and neonatal/infant death is significantly higher among twins with MC compared to DC placentation. Twins conceived by AR have a much lower risk of MC placentation. The risk of losing one or both twins seems higher among naturally conceived twins compared to twins conceived by AR, despite the fact that the maternal age was higher among the mothers of the AR twins.

Adult↗
PubMed2005

How to identify twins at low risk of spontaneous preterm delivery.

OBJECTIVE: The aim of this study was to evaluate transvaginal sonographic assessment of cervical length at 23 weeks as a screening test for spontaneous preterm delivery in order to define a cut-off value that could be used to select twin pregnancies at low risk of spontaneous preterm delivery. METHODS: In a prospective multicenter study of 383 twin pregnancies included before 14 + 6 weeks a cervical scan with measurement of the cervical length was performed at 23 weeks' gestation. The results were blinded for the clinicians if the cervical length was > or = 15 mm. The rates of spontaneous delivery at different cut-off levels of cervical length were determined. RESULTS: Eighty-nine percent of the twins had dichorionic placentation and 58% were conceived after assisted reproduction. The rate of spontaneous preterm delivery was 2.3% (1.5% for dichorionic (DC) and 9.1% for (MC) monochorionic twins) before 28 weeks and 18.5% (17.1% for DC and 29.5% for MC twins) before 35 weeks. The screen-positive rate was 5% for a cervical length < or = 20, 7-8% at < or = 25, 16-17% at < or = 30 and 34-48% at < or = 35 mm depending on chorionicity. The false-negative rate (1 - negative predictive value) ranged from 1.2% at 28 weeks to 18.6% at 35 weeks for all twins. Receiver-operating characteristics curves showed that the sensitivity increased with declining gestational age with cut-off levels of highest accuracy at 21 mm for 28 weeks and 29 mm for 33 weeks. CONCLUSIONS: Cervical length measurement at 23 weeks of gestation is a good screening test for predicting twins at low risk of preterm and very preterm delivery, especially in DC twins. The present results suggest that a cut-off of 25 mm should be recommended.

Cervical Ripening↗
PubMed2003

Update on cicatricial alopecia.

Cicatricial alopecia is an enigmatic group of hair disorders linked by the potential permanent loss of scalp hair follicles in involved areas. Progress in our understanding and treatment of these disorders has been stymied by the lack of clear diagnostic criteria for the current terms used to describe the various hair loss entities. Since all of these conditions evolve as the hair is destroyed or replaced, diagnosis is further made difficult by a lack of clinical and pathologic "snapshots" over the evolution of each disorder. Without some acceptance of general clinical and histological presentations in the early, mid and late stage of these disorders, one cannot begin to explore ways to make the diagnosis at a very early stage before significant follicular destraction has occurred (making the clinical diagnosis obvious) and when the damage is potentially repairable or progression preventable.

Alopecia↗
PubMed2001

Transgene-mediated post-transcriptional gene silencing is inhibited by 3' non-coding sequences in Paramecium.

Homology-dependent gene silencing is achieved in Paramecium by introduction of gene coding regions into the somatic nucleus at high copy number, resulting in reduced expression of all homologous genes. Although a powerful tool for functional analysis, the relationship of this phenomenon to gene silencing mechanisms in other organisms has remained obscure. We report here experiments using the T4a gene, a member of the trichocyst [corrected]matrix protein (TMP) multigene family encoding secretory proteins, and the ND7 gene, a single copy gene required for exocytotic membrane fusion. Silencing of either gene leads to an exocytosis-deficient phenotype easily scored on individual cells. For each gene we have tested the ability of different combinations of promoter, coding and 3' non-coding regions to provoke silencing, and analyzed transcription and steady-state RNA in the transformed cells. We provide evidence that homology-dependent gene silencing in Paramecium is post-transcriptional and that both sense and antisense RNA are transcribed from the transgenes, consistent with a role for dsRNA in triggering silencing. Constructs with and without promoters induce gene silencing. However, transgenes that contain 3' non-coding regions do not induce gene silencing, despite antisense RNA production. We present a model according to which different pathways of RNA metabolism compete for transcripts and propose that the relative efficiencies of dsRNA formation and of 3' RNA processing of sense transgene transcripts determine the outcome of transformation experiments.

3' Untranslated Regions↗
PubMed2001

Paramecium genome survey: a pilot project.

A consortium of laboratories undertook a pilot sequencing project to gain insight into the genome of Paramecium. Plasmid-end sequencing of DNA fragments from the somatic nucleus together with similarity searches identified 722 potential protein-coding genes. High gene density and uniform small intron size make random sequencing of somatic chromosomes a cost-effective strategy for gene discovery in this organism.

Animals↗
PubMed2001

Genetic interference in protozoa.

RNA interference first described in Caenorhabditis elegans and transgene-induced post-transcriptional gene silencing first described in plants and fungi now appear as different means of activating a conserved and ancient mechanism that can protect genomes against viruses and transposons and perhaps also control expression of endogenous genes. We present here similar genetic interference phenomena in highly divergent protozoa, Trypanosoma and Paramecium, and look ahead to what contribution these microorganisms could bring to this fast-moving area.

Animals↗
PubMed2001

Twin pregnancy: the role of ultrasound in management.

Determination of chorionicity is one of the most important issues in the management of twin pregnancy. Modern ultrasound equipment has made it possible to accurately assess placentation already in the first trimester with the lambda sign. With regard to prenatal diagnosis, it is important to know the chorionicity in order to calculate the risk of chromosomally abnormal fetuses. Accurate chorionicity offers the obstetricians the opportunity to observe the monochorionic twins more intensively than is required for twins with dichorionic placentation. This review gives an update of the state of the art for clinicians caring for twin pregnancies.

Amniocentesis↗
PubMed2001

Growth and form of secretory granules involves stepwise assembly but not differential sorting of a family of secretory proteins in Paramecium.

Paramecium trichocysts are voluminous secretory vesicles consisting of a spindle-shaped body surmounted by a tip that serves to anchor them at exocytotic sites in the plasma membrane. This constrained shape is conferred by the proteins stored in the vesicles, which form an insoluble three-dimensional crystalline array. The constituent polypeptides (Trichocyst Matrix Proteins, TMPs), which assemble during trichocyst biogenesis, are produced by proteolytic processing of soluble proproteins encoded by a large multigene family. In order to investigate the functional significance of the TMP multigene family, which assures the synthesis of a mixture of related polypeptides, we have designed synthetic genes for heterologous expression of three different mature polypeptides, which were used to obtain sequence-specific rabbit antisera. We used these antisera to carry out immunolocalization experiments with wild-type trichocysts at different stages of development and found that the trichocyst matrix consists of two concentric layers containing different TMPs, and that the assembly of each layer corresponds to a distinct phase of trichocyst growth. Examination of mutant trichocysts created by targeted gene silencing of different TMP genes showed that the layer containing the products of the silenced genes is specifically affected, as are all subsequently assembled parts of the structure, consistent with an ordered assembly pathway. This stepwise assembly is not controlled by differential sorting of the TMPs, as single and double label experiments provided evidence that the different TMPs are delivered together to post-Golgi vesicles and developing trichocysts. We present a model for trichocyst biogenesis in which TMP assembly is controlled by protein processing.

Amino Acid Sequence↗
PubMed2000

Molecular genetics of regulated secretion in paramecium.

Paramecium is a unicell in which cellular processes are amenable to genetic dissection. Regulated secretion, which designates a secretory pathway where secretory products are first stored in intracellular granules and then released by exocytotic membrane fusion upon external trigger, is an important function in Paramecium, involved in defensive response through the release of organelles called trichocysts. In this review, we focus on recent advances in the molecular genetics of two major aspects of the regulated pathway in Paramecium, the biogenesis of the secretory organelles and their exocytosis.

Animals↗
PubMed2000

Regulation of secretory protein gene expression in paramecium role of the cortical exocytotic sites.

In cells that possess a regulated secretory pathway, exocytosis can lead to transcriptional activation of genes encoding products stored in secretory granules as well as genes required for granule biogenesis. With the objective of understanding this response, we have examined the expression of Paramecium secretory protein genes in different physiological and genetic contexts. The genes belong to the trichocyst matrix protein (TMP) multigene family, encoding polypeptides that form the crystalline matrix of the secretory granules, known as trichocysts. Approximately 1000 trichocysts per cell are docked at pre-formed cortical exocytotic sites. Their rapid and synchronous exocytosis can be triggered by vital secretagogues such as aminoethyldextran without harming the cells. Using this exocytotic trigger, we found that the transcription of TMP genes undergoes rapid, transient and co-ordinate 10-fold activation in response to massive exocytosis, leading to a 2.5-fold increase in the pool of TMP mRNA. Experiments with exocytosis-deficient mutants show that the secretagogue-induced increase in intracellular free calcium implicated in stimulus/secretion coupling is not sufficient to activate TMP gene expression. We present evidence that the state of occupation of the cortical exocytotic sites can affect TMP gene expression and suggest that these sites play a role in gene activation in response to exocytosis.

Animals↗
PubMed1999

The effect of decreasing digital image resolution on teledermatology diagnosis.

OBJECTIVE: To determine the effect of degraded digital image resolution (as viewed on a monitor) on the accuracy and confidence of dermatologic interpretation. MATERIALS AND METHODS: Eight dermatologists interpreted 180 clinical cases divided into three Logical Competitor Sets (LCS) (pigmented lesions, non-pigmented lesions, and inflammatory dermatoses). Each case was digitized at three different resolutions. The images were randomized and divided into (9) 60-image sessions. The physicians were completely blinded concerning the image resolution. After 60 seconds per image, the viewer recorded a diagnosis and level of confidence. The resultant ROC curves compared the effect of LCS, level of clinical difficulty, and resolution of the digital image. One-way analysis of variance (ANOVA) compared the curves. RESULTS: The areas beneath the ROC curves did not demonstrate any consistently significant difference between the digital image resolutions for all LCS and levels of difficulty. The only significant effect observed was amongst pigmented lesions (LCS-A) where the ROC curve area was significantly smaller in the easy images at high resolution compared to low and medium resolutions. For all other ROC curve comparisons within LCS-A, at all other levels of difficulty, as well as within the other LCS at all levels of difficulty, none of the differences was significant. CONCLUSION: A 720 x 500 pixel image can be considered equivalent to a 1490 x 1000 pixel image for most store-and-forward teledermatology consultations.

Analysis of Variance↗
PubMed1998

Homology-dependent gene silencing in Paramecium.

Microinjection at high copy number of plasmids containing only the coding region of a gene into the Paramecium somatic macronucleus led to a marked reduction in the expression of the corresponding endogenous gene(s). The silencing effect, which is stably maintained throughout vegetative growth, has been observed for all Paramecium genes examined so far: a single-copy gene (ND7), as well as members of multigene families (centrin genes and trichocyst matrix protein genes) in which all closely related paralogous genes appeared to be affected. This phenomenon may be related to posttranscriptional gene silencing in transgenic plants and quelling in Neurospora and allows the efficient creation of specific mutant phenotypes thus providing a potentially powerful tool to study gene function in Paramecium. For the two multigene families that encode proteins that coassemble to build up complex subcellular structures the analysis presented herein provides the first experimental evidence that the members of these gene families are not functionally redundant.

Animals↗
PubMed1997

Characterization of multigene families in the micronuclear genome of Paramecium tetraurelia reveals a germline specific sequence in an intron of a centrin gene.

In Paramecium, as in other ciliates, the transcriptionally active macronucleus is derived from the germline micronucleus by programmed DNA rearrangements, which include the precise excision of thousands of germline-specific sequences (internal eliminated sequences, IESs). We report the characterization of micronuclear versions of genes encoding Paramecium secretory granule proteins (trichocyst matrix proteins, TMPs) and Paramecium centrins. TMP and centrin multigene families, previously studied in the macronuclear genome, consist of genes that are co-expressed to provide mixtures of related polypeptides that co-assemble to form respectively the crystalline trichocyst matrix and the infraciliary lattice, a contractile cytoskeletal network. We present evidence that TMP and centrin genes identified in the macronucleus are also present in the micronucleus, ruling out the possibility that these novel multigene families are generated by somatic rearrangements during macronuclear development. No IESs were found in TMP genes, however, four IESs in or near germline centrin genes were characterized. The only intragenic IES is 75 bp in size, interrupts a 29 bp intron and is absent from at least one other closely related centrin gene. This is the first report of an IES in an intron in Paramecium.

Amino Acid Sequence↗
PubMed1996

Cloning and sequence analysis of genes coding for paramecium secretory granule (trichocyst) proteins. A unique protein fold for a family of polypeptides with different primary structures.

The architecturally complex secretory granules of Paramecium, known as trichocysts, have two unusual and seemingly contradictory features: their protein contents have crystalline organization (Sperling, L., Tardieu, A., and Gulik-Krzywicki, T. (1987) J. Cell Biol. 105, 1649-1662), yet these proteins are a heterogeneous set of molecules encoded by a large multigene family (Madeddu, L., Gautier, M.-C., Vayssié, L., Houari, A., and Sperling, L. (1995) Mol. Biol. Cell 6, 649-659). We present here the first complete sequences of three genes coding for three different precursors of the trichocyst crystalline matrix proteins. The deduced protein sequences indicate that each precursor gives rise to two of the mature polypeptides found in the crystalline trichocyst matrix. Analysis of putative processing sites suggests that a series of reactions, some of which may involve a novel endopeptidase, are involved in their proteolytic maturation. Each of the 6 mature polypeptides contains heptad segments. Characterization of the heptad segments leads us to propose that the mature polypeptides that compose the crystalline trichocyst matrix, despite their different primary structures, all share a unique protein fold, probably a 4 alpha-helical antiparallel bundle.

Amino Acid Sequence↗
PubMed1996

Clinical and histopathologic features of hair loss in patients with HIV-1 infection.

BACKGROUND: Hair loss is common in patients with HIV-1 infection, and in black patients this loss may be associated with straightening. Possible causes are frequently present in patients with HIV-1. These causes include chronic HIV-1 infection itself and recurrent secondary infections, nutritional deficiencies, immunologic and endocrine dysregulation, and exposure to multiple drugs. However, histopathologic features have rarely been reported in these patients. OBJECTIVE: The objective was to evaluate the changes in the hairs of a group of these patients and to identify the light microscopic and ultrastructural changes in the hairs and the histologic changes in the scalp. METHODS: Hair plucks and pulls with scanning electron microscopy of the hairs were done on 10 patients with late-stage HIV-1 infection. In addition, scalp biopsy specimens were examined in both vertical and transverse sections. RESULTS: All patients had telogen effluvium. Numerous apoptotic or necrotic keratinocytes were seen in the upper external root sheath follicular epithelium in addition to a mild to moderate perifollicular mononuclear cell infiltrate often containing eosinophils. Variable dystrophy of the hair shafts was also a consistent feature. CONCLUSION: Although telogen effluvium is a common response to a wide spectrum of biologic stresses, the presence of apoptotic or necrotic keratinocytes within the upper end of the external root sheath epithelium and dystrophy of hairs may be markers of hair loss in patients with HIV-1 infection.

Acquired Immunodeficiency Syndrome↗
PubMed1995

Ergonomic development work: co-education as a support for user participation at a car assembly plant. A case study.

This study deals with the design, trials and evaluation of a co-education programme at the Volvo Uddevalla plant in Sweden. Involving operators, manufacturing engineers and managers, the programme served as a support for the creation of a participatory ergonomics process, intended for continuous use at the plant. It consisted of a basic ergonomics knowledge package, and a dialogue model defining the roles and relations of actors involved. As a practical part of the programme, trial development projects were also carried out by the participants. The main and long term objective of the project was to start the participants cooperating in a continuous change and development process on the shop-floor. The outcome of the co-education programme was evaluated immediately after the first two regular courses, and, as a longterm follow-up, after seven subsequent courses shortly after the closing of the Uddevalla plant. The co-education programme was shown to be successful. Later on, the expertize of both operators and manufacturing engineers became obvious to everyone at the plant, and the cooperation between operators and manufacturing engineers increased steadily. The main conclusion drawn was that the co-education programme is a good starting point for a process of participation and industrial change work. However, in order to get a permanent impact, the whole organization must nurse and nourish the further development, and implementation of the process.

Journal Article↗