Mapping Human Fetal Brain Maturation In Vivo Using Quantitative MRI.

BACKGROUND AND PURPOSE: On the basis of a single multidynamic multiecho sequence acquisition, SyMRI generates a variety of quantitative image data that can characterize tissue-specific properties. The aim of this retrospective study was to evaluate the feasibility of SyMRI for the qualitative and quantitative assessment of fetal brain maturation. MATERIALS AND METHODS: In 52 fetuses, multidynamic multiecho sequence acquisitions were available. SyMRI was used to perform multidynamic multiecho-based postprocessing. Fetal brain maturity was scored qualitatively on the basis of SyMRI-generated MR imaging data. The results were compared with conventionally acquired T1-weighted/T2-weighted contrasts as a standard of reference. Myelin-related changes in T1-/T2-relaxation time/relaxation rate, proton density, and MR imaging signal intensity of the developing fetal brain stem were measured. A Pearson correlation analysis was used to detect correlations between the following: 1) the gestational age at MR imaging and the fetal brain maturity score, and 2) the gestational age at MR imaging and the quantitative measurements. RESULTS: SyMRI provided images of sufficient quality in 12/52 (23.08%) (range, 23 + 6-34 + 0) fetal multidynamic multiecho sequence acquisitions. The fetal brain maturity score positively correlated with gestational age at MR imaging (SyMRI: r = 0.915, P < .001/standard of reference: r = 0.966, P < .001). Myelination-related changes in the T2 relaxation time/T2 relaxation rate of the medulla oblongata significantly correlated with gestational age at MR imaging (T2-relaxation time: r = -0.739, P = .006/T2-relaxation rate: r = 0.790, P = .002). CONCLUSIONS: Fetal motion limits the applicability of multidynamic multiecho-based postprocessing. However, SyMRI-generated image data of sufficient quality enable the qualitative assessment of maturity-related changes of the fetal brain. In addition, quantitative T2 relaxation time/T2 relaxation rate mapping characterizes myelin-related changes of the brain stem prenatally. This approach, if successful, opens novel possibilities for the evaluation of structural and biochemical aspects of fetal brain maturation.

Strategies for the evaluation and selection of potential vaginal probiotics from human sources: an exemplary study.

During the last years, the application of probiotics in gynaecological clinical practice has gained increasing relevance regarding therapy and prevention. This trend has also provoked the need for having tailored pharmaceutical preparations containing powerful microbial strains with defined properties. For the development of such preparations, several factors and criteria have to be considered, thereby not only focusing on identity and safety aspects as well as individual properties of the bacterial strains, but also on technological issues, such as stability and targeted release from the preparation. Against this background, this report exemplarily addresses the development procedure of a probiotic bacterial formulation for gynaecological application, covering the search for suitable strains, assessing their microbiological, molecular biological and physiological characterisation, and the selection for their use in clinical trials. In detail, starting with 127 presumptive lactobacilli isolates of vaginal origin, a step-by-step selection of candidate strains meeting special criteria was thoroughly examined, finally leading to a preparation consisting of four individual Lactobacillus strains that possess particular significance in women's urogenital health. Relevant issues and quality criteria of probiotic preparations used in gynecology are addressed and exemplarily introduced.

Differences in the vaginal lactobacilli of postmenopausal women and influence of rectal lactobacilli.

OBJECTIVE: This study was undertaken to characterize the Lactobacillus spp. dominating the vaginal microbiota of healthy postmenopausal women and to determine the possible influence of rectal lactobacilli. METHODS: Sixty postmenopausal women aged 55-65 years without clinical signs of vaginal infection not receiving hormone replacement therapy were included in this cross-sectional observational study. Based on Gram-stained smears, 30 women with a normal vaginal flora (Nugent score 0) were included in Group 1, and 30 women with an intermediate vaginal flora characterized by an absence of vaginal lactobacilli (Nugent score 4) were included in Group 2. Vaginal and rectal smears were taken for molecular lactobacillus profiling using polymerase chain reaction and denaturing gradient gel electrophoresis. Diversity of vaginal and rectal lactobacilli in postmenopausal women was the main outcome measure. RESULTS: We noticed a minor interference of gut lactic acid bacteria on a normal vaginal microflora dominated by lactobacilli strains of the L. delbrueckii group. When the normal vaginal microflora is disturbed by depletion of lactobacilli, the gut may function as a reservoir for lactobacilli of the L. casei group, which then colonize the vagina. CONCLUSION: Our data indicate that rectal lactobacilli may affect the vaginal flora of postmenopausal women in the case of lactobacillary absence and help to maintain a normal vaginal microbiota.

Monthly itraconazole versus classic homeopathy for the treatment of recurrent vulvovaginal candidiasis: a randomised trial.

OBJECTIVE: Antimycotics effectively treat sporadic and recurrent vulvovaginal candidiasis (RVVC). Classic homeopathy (CH) is also used to treat this condition. We compared the efficacy of CH and itraconazole in reducing the frequency of RVVC episodes. DESIGN: Single-centre, prospective, randomised trial. SAMPLE: One hundred-and-fifty patients with a history of RVVC and an acute episode of VVC. METHODS: Women were randomised into 3 groups: itraconazole with lactobacilli (group 1), itraconazole without lactobacilli (group 2) and CH (group 3). Itraconazole treatment of acute infection was followed by a 6-month maintenance regimen with monthly single-day itraconazole (200 mg bid). Women in group 1 were given additional vaginal lactobacilli for 6 days per month throughout the maintenance regimen Thereafter, patients were followed without treatment for 6 months. CH treatment was performed for 12 months. RESULTS: Women in groups 1 and 2 reached a culture-free status significantly earlier than women in group 3 (log-rank test; P < 0.0001). Specifically, before the start of the maintenance regimen, 44 of 49 women (89.8%) in group 1 and 40 of 47 women (85%) in group 2 were free of Candida detectable by culture, 22 of 46 (47%) women in group 3 reached a culture-free status after the first visit, but had a recurrence significantly earlier compared with women in groups 1 and 2 (log-rank test; P = 0.002). After 12 months, 19 of 25 (76%) women in group 1, 18 of 23 (78%) women in group 2 and 9 of 23 (39%) women in group 3 were free of culture-detectable Candida. Assessment of RVVC-associated complaints by VAS score showed that women in group 3 had a significantly higher level of discomfort (36.8, 25.1 and 27.7 respectively; P < 0.001) and were significantly less satisfied (59.2, 68.2 and 71.7 respectively; P < 0.001) than patients in groups 1 and 2. CONCLUSIONS: Monthly cycle-dependent itraconazole is more effective than CH in the treatment of RVVC. Lactobacilli do not confer an added benefit.

Vaginal Lactobacillus microbiota of healthy women in the late first trimester of pregnancy.

OBJECTIVE: This study was undertaken to characterise the dominant species of Lactobacillus colonising the vagina of healthy pregnant women, to examine some of their phenotypic and genotypic properties, and to gain a better understanding of the potential role of species, which might be associated with infection-free status. DESIGN: A prospective descriptive cohort study. SETTING: Department of Obstetrics and Gynaecology, Medical University of Vienna and Medical School, Vienna, Austria. SAMPLE: A total of 200 women in the late first trimester of pregnancy without clinical signs of vaginal infection were included in the study. Of these, 126 women were found to have a normal vaginal flora based on Gram stain. METHODS: Culture probes from those 126 women were further processed for identification of Lactobacillus species. Overall, 168 colonies from 84 women were identified as belonging to the Lactobacillus genus. Based on the combined results of microbiological methods and genus-specific, multiplex, and species-specific polymerase chain reaction, lactobacilli were recovered from 72 women. MAIN OUTCOME MEASURES: Identification of Lactobacillus species of the vaginal flora of healthy pregnant women. RESULTS: The most frequently occurring species were Lactobacillus crispatus and Lactobacillus gasseri, followed by Lactobacillus jensenii and Lactobacillus rhamnosus. CONCLUSIONS: Our results may have implications on the composition and on the use of Lactobacillus preparations for the prevention of recurrent vaginal infection.

Cost effectiveness of a screen-and-treat program for asymptomatic vaginal infections in pregnancy: towards a significant reduction in the costs of prematurity.

OBJECTIVE: The purpose of this investigation was to determine the cost-saving potential of a simple screen-and-treat program for vaginal infection, which has previously been shown to lead to a reduction of 50% in the rate of preterm births. STUDY DESIGN: To determine the potential cost savings, we compared the direct costs of preterm delivery of infants with a birth weight below 1900g with the costs of the screen-and-treat program. We used a cut-off birth weight of 1900g because, in our population, all infants with a birth weight below 1900g were transferred to the neonatal intensive care unit. The direct costs associated with preterm delivery were defined to include the costs of the initial hospitalization of both mother and infant and the costs of outpatient follow-up throughout the first 6 years of life of the former preterm infant. The costs of the screen-and-treat program were defined to include the costs of the screening examination and the resulting costs of antimicrobial treatment and follow-up. All calculations were based on health-economic data obtained in the metropolitan area of Vienna, Austria. RESULTS: The number of preterm infants with a birth weight below 1900g was 12 (0.5%) in the intervention group (N=2058) and 29 (1.3%) in the control group (N=2097). The direct costs per preterm birth were found to amount to EUR (euro) 60262. Overall, the expected total savings in direct costs achieved by the screen-and-treat program and the ensuing 50% reduction in the number preterm births with a birth weight below 1900g amounted to more than euro 11 million. The costs of screening and treatment were found to amount to merely 7% of the direct costs saved as a result of the screen-and-treat program. CONCLUSION: A simple preterm prevention program, consisting of screening and antimicrobial treatment and follow-up of women with asymptomatic vaginal infection, leads not only to a significant reduction in the rate of preterm births but also to substantial savings in the direct costs associated with prematurity.

Evolutionarily plastic regions at human 3p21.3 coincide with tumor breakpoints identified by the "elimination test".

We have previously found with the microcell hybrid-based "elimination test" that human chromosome 3 transferred into murine or human tumor cells regularly lost certain 3p regions during tumor growth in SCID mice. The most common eliminated region, CER1, is approximately 2.4 Mb at 3p21.3. CER1 breakpoints were clustered in approximately 200-kb regions at both telomeric and centromeric borders. We have also shown, earlier, that tumor-related deletions often coincide with human/mouse synteny breakpoints on 3p12-p22. Here we describe the results of a comparative genomic analysis on the CER1 region in Caenorhabditis elegans, Drosophila melanogaster, Fugu rubripes, Gallus gallus, Mus musculus, Rattus norvegicus, and Canis familiaris. First, four independent synteny breaks were found within the CER1 telomeric breakpoint cluster region, comparing human, dog, and chicken genomes, and two independent synteny breaks within the CER1 centromeric breakpoint cluster region, comparing human, mouse, and chicken genomes, suggesting a nonrandom involvement of tumor breakpoint regions in chromosome evolution. Second, both CER1 breakpoint cluster regions show recent tandem duplications (seven Zn finger protein family genes at the telomeric and eight chemokine receptor genes at the centromeric side). Finally, all genes from these regions underwent horizontal evolution in mammals, with formation of new genes and expansion of gene families, which were displayed in the human genome as tandem gene duplications and pseudogene insertions. In contrast the CER1 middle region contained evolutionarily well-conserved solitary genes and a minimal amount of retroposed genes. The coincidence of evolutionary plasticity with CER1 breakpoints may suggest that regional structural instability is expressed in both evolutionary and cancer-associated chromosome rearrangements.

The LZTFL1 gene is a part of a transcriptional map covering 250 kb within the common eliminated region 1 (C3CER1) in 3p21.3.

Deletions on 3p have been described in a large number of human tumors, suggesting the presence of a tumor suppressor gene(s). Using the elimination test, we previously defined a 1-Mb segment from human 3p21.3 (C3CER1). Genomic sequencing allowed us to construct a transcription map covering 250 kb containing five genes. We have characterized a human leucine zipper containing gene, leucine zipper transcription factor-like 1 (LZTFL1), and its mouse orthologue (Lztfl1), which was also mapped to mouse chromosome 9F. The LZTFL1 gene has two transcript isoforms displaying alternative polyadenylation. We have localized the human orthologue of the yeast SAC1 (suppressor of actin) gene as well as characterized and mapped the mouse Sac1 gene. Furthermore, the XT3 gene was characterized, encoding a member of the Na(+)/Cl(-) neurotransmitter superfamily. It has been shown that the XT3 gene had an alternatively spliced brain-specific isoform, predicted to remove 1 of 12 putative transmembrane domains. The transcription map also includes the CC chemokine receptor 9 gene (CCR9) and the LIM domain containing gene 1 (LIMD1). This work partially defines the gene content of C3CER1 that is a prerequisite for delineation of its role in tumorigenesis.

Postoperative aneurysm of the popliteal artery after arthroscopic meniscectomy.

We report a case of a 14-year-old girl who underwent arthroscopic subtotal meniscectomy after a bucket-handle tear of the lateral meniscus followed by a pseudoaneurysm of the popliteal artery. A revision surgery and reconstruction of the artery with a saphena magna patch was necessary to repair the defect. At 1-year follow-up, excellent range of motion, function of the knee, and a normal quality of life was achieved.

Similar regions of human chromosome 3 are eliminated from or retained in human/human and human/mouse microcell hybrids during tumor growth in severe combined immunodeficient (SCID) mice.

By passaging microcell hybrids (MCHs) containing human chromosome 3 (chr3) on A9 mouse fibrosarcoma background through severe combined immunodeficient (SCID) mice (elimination test), we have previously defined a 1-Mb-long common eliminated region 1 (CER1) at 3p21.3, a second eliminated region (ER2) at 3p21.1-p14 and a common retained region (CRR) at 3q26-qter. In the present work, chr3 was transferred by microcell fusion into the human nonpapillary renal cell carcinoma line KH39 that contained uniparentally disomic chr3. Four MCHs were generated. Compared with KH39, they developed fewer and smaller tumors, which grew after longer latency periods in SCID mice. The tumors were analyzed in comparison with corresponding MCHs by chr3 arm-specific painting, 19 fluorescent in situ hybridization (FISH) probes, and 27 polymorphic markers. Three MCHs that maintained the intact exogenous chr3 in vitro lost one 3p copy in all 11 tumors. Seven of 11 tumors lost the exogenous 3p, whereas four tumors contained mixed cell populations that lacked either the exogenous or one endogenous KH39 derived 3p. In one MCH the exogenous chr3 showed deletions within CER1 and ER2 already in vitro. It remained essentially unchanged in 8/9 derived tumors. The third, exogenous copy of the 3q26-q27 region (part of CRR) was retained in 16/20 tumors. It can be concluded that the human/human MCH-based elimination test identifies similar eliminated and retained regions on chr3 as the human/murine MCH-based test.

Development of the gametophytes, flower, and floral vasculature in Dichorisandra thyrsiflora (Commelinaceae).

The flowers of Dichorisandra thyrsiflora (Commelinaceae) are monosymmetric and composed of three sepals, three petals, six stamens, and three connate carpels. The anthers are poricidal and possess a wall of five cell layers (tapetum included). This type of anther wall, not previously observed in the Commelinaceae, is developmentally derived from the monocotyledonous type via an additional periclinal division and the persistence of the middle layers through anther dehiscence. Secondary endothecial thickenings develop in the cells of the two middle layers only. The tapetum is periplasmodial and contains raphides. Microsporogenesis is successive and yields both decussate and isobilateral tetrads. Pollen is shed as single binucleate grains. The gynoecium is differentiated into a globose ovary, hollow elongate style, and trilobed papillate stigma. Each locule contains six to eight hemianatropous to slightly campylotropous crassinucellar ovules with axile (submarginal) placentation. The ovules are bitegmic with a slightly zig-zag micropyle. Megagametophyte development is of the Polygonum type. The mature megagametophyte consists of an egg apparatus and fusion nucleus; the antipodals having degenerated. The floral vasculature is organized into an outer and inner system of bundles in the pedicel. The outer system becomes ventral carpellary bundles. All other floral vascular traces originate from the inner system.

Increased blood pressure response to the cold pressor test in pregnant women developing pre-eclampsia.

OBJECTIVES: Recent data indicate an increased vascular reactivity due to an overactivity of the sympathetic nervous system in women with pre-eclampsia. We therefore evaluated whether this increased vascular reactivity can be detected prior to the clinical manifestation of preeclampsia by the use of a physiological stimulus. DESIGN: Prospective data collection. SETTING: Clinic of Obstetrics and Gynecology in a 2000 bed tertiary care hospital. PARTICIPANTS: One hundred and twenty-three pregnant women between the 16th to 20th week of gestation. INTERVENTIONS: A cold pressor test was performed by positioning an ice-bag on the forehead of the woman for 3 min. Blood pressure and heart rate were monitored by a continuous, noninvasive blood pressure measurement device during the stimulus and after removal of the icebag. A clinical follow-up was carried out by review of the charts after delivery to identify those women who have developed pre-eclampsia. RESULTS: Ten (8%) out of 123 pregnant women developed pre-eclampsia. During the cold pressor test systolic as well as diastolic blood pressure increased significantly and was more pronounced in women developing pre-eclampsia compared with healthy pregnant women (systolic blood pressure: 14.2 +/- 5.5 versus 8.5 +/- 7.2 mmHg, P= 0.02; diastolic blood pressure: 7.3 +/- 4.9 versus 3.9 +/- 4.7 mmHg, P=0.03). The change in heart rate was similar between both groups (8 +/- 2.6 versus 10.4 +/- 6.4 beats/min, not significant). CONCLUSIONS: An increased vasoconstrictive response to a physiological stimulus is present in women with pre-eclampsia as a sign of an increased vascular reactivity prior to clinical manifestation of the disease. The cold pressor test may be a suitable diagnostic tool to identify women, who will develop pre-eclampsia. However, future studies in larger cohorts are required to establish the final value of this test.

Fetal fibronectin as a predictor of term labor: a literature review.

OBJECTIVE: Accurately predicting the onset of delivery continues to be problematic notwithstanding the clinical relevance of this capability, especially with regard to the elective induction of labor. While the assessment of the cervix according to Bishop, to date, constitutes the single generally recognized method, the determination of fetal fibronectin (FFN) cervicovaginal secretions promises greater precision in this context. DESIGN: English-language medical literature was analyzed using the search parameter 'fetal fibronectin', 'term delivery', 'induction of labor', 'labor', and 'birth', respectively. All those original reports examining cervicovaginal FFN that were originally published in English and followed a prospective design were subsequently included in the study at hand. These reports were evaluated in regard to their findings on the predictive value of FFN for spontaneous delivery and induction of labor. RESULTS: A total of eight reports were included in the present study: five of these, covering a total of 480 patients, discussed the relationship between FFN and the spontaneous onset of labor; the other three addressed the subject of FFN and the induction of labor (300 patients). All reports clearly related the presence of FFN in cervicovaginal secretions to the imminent onset of delivery; three studies, furthermore, demonstrated a correlation with the spontaneous onset of labor, with a sensitivity greater than 90%. In case of the studies concerning induction of labor, a positive FFN result was accompanied by a significantly shorter interval until delivery than a negative FFN result. CONCLUSION: Determining the presence of FFN in cervicovaginal secretions promises to provide an indicator relating to the prediction of term labor, as well as to the prediction of successful labor induction.

How can plants tell which way is up?

Many people think of plants as essentially sessile organisms that do not actively respond to their environment. What could be further from the truth! In fact, plants are capable of a variety of movements, including the dramatic nastic responses (such as Venus fly trap closure) and the less sensational tropisms. These latter movements are directed growth responses to some type of external stimulus such as gravity (gravitropism, formerly known as geotropism) or light (phototropism). This paper describes some interesting exercises that are derived from recent work, including research that has led to experiments performed on two Space Shuttle missions in 1997 (Kiss et al. 1998). The study of tropisms can be a useful way to introduce students to plant biology in high school and introductory college courses. In our experience, students are fascinated by plant movements when they are presented in lectures and find laboratory experiences on this topic quite engaging. Laboratory work on plant tropisms can also be used to introduce important concepts in science such as hypothesis testing, quantitative analysis, and the use of statistics. The laboratory exercises described in this paper involve the higher plant Arabidopsis thaliana, which has become an important organism in molecular biology research and is the focus of an international plant genome project. Based on the material presented here, a number of plant gravitropism laboratory exercises with Arabidopsis that are simple in terms of equipment/materials and procedures can be developed. These exercises are robust in that they work well even in the hands of introductory students, and they can be expanded according to the individual instructor's needs. This paper describes two exercises that have been performed by beginning college students, and these exercises can easily be performed in biology classes in most high school settings.

A 1-Mb PAC contig spanning the common eliminated region 1 (CER1) in microcell hybrid-derived SCID tumors.

We have developed an elimination test to identify chromosomal regions that contain tumor inhibitory genes. Monochromosomal human/mouse microcell hybrids are generated and passaged through SCID mice. Derived tumors are then analyzed for deletions on the transgenomic chromosome. Using this strategy, we have previously identified a 1.6-cM common eliminated region 1 (CER1) on human 3p21. 3. We now report that CER1 contains 14 markers that are deleted in 19 SCID-derived tumors. A 1-Mb PAC contig that spans CER1 was assembled. Five chemokine receptor genes (CCR1, CCR3, CCR2, CCR5, and CCR6) were localized in CER1 in a 225-kb cluster. The lactotransferrin gene (LTF, or lactoferrin, LF), which reportedly has tumor inhibitory activity, also maps to CER1. Our results create a basis for characterization and further functional testing of genes within CER1.

A novel gene containing LIM domains (LIMD1) is located within the common eliminated region 1 (C3CER1) in 3p21.3.

Chromosomal deletions on 3p have been described in a large number of human tumors, suggesting the presence of a tumor suppressor gene(s). Using an experimental system, called the elimination test, we previously identified a 1 Mb segment, the common eliminated region 1 (C3CER1). C3CER1 was also covered by a PAC contig. Using the sequence of two overlapping PACs from C3CER1, we localized the human KIAA0028 cDNA, encoding the precursor of mitochondrial leucyl-tRNA synthetase. We also characterized a novel human LIM domain-containing gene (LIMD1) and its mouse ortholog (Limd1). LIM domains consist of a cysteine-rich consensus sequence containing two distinct zinc-binding subdomains, which mediate protein-protein interactions. The predicted protein sequences of the human and mouse genes reveal three LIM domains located at the C-terminal end, which indicates that they belong to the group 3 of the gene family encoding LIM motifs. We characterized the genomic structure of the human LIMD1 gene and assigned the mouse Limd1 gene to the chromosome 9F subtelomeric region. Both genes are ubiquitously expressed at the mRNA level. The LIM motif has been previously identified in many developmentally important factors from various eukaryotes. These factors have been shown to play a role in intracellular signaling, transcriptional regulation and cellular differentiation during development. The human C3CER1-located LIMD1 gene should therefore be further studied for its possible role in tumor suppression.

Expression of endothelial (type III) nitric oxide synthase in cytotrophoblastic cell lines: regulation by hypoxia and inflammatory cytokines.

Expression of endothelial nitric oxide synthase (eNOS) has been localized to the villous syncytiotrophoblasts suggesting that NO release from these cells could prevent platelet adhesion and aggregation in the intervillous space. Hypoxia- or inflammation-dependent changes in the release of this vasoactive substance may result in thrombus formation and altered vascular resistance which occur in the placental bed of pre-eclamptic patients. To evaluate the influence of low-oxygen tension and inflammation on eNOS production in the trophoblast steady-state eNOS mRNA and protein levels were investigated in cytotrophoblastic BeWo and Jeg-3 cells cultured at 3.5 per cent oxygen and/or in the presence of the pro-inflammatory cytokines IL-1 and TNF-alpha. By RT-PCR and immunocytochemistry we demonstrate that BeWo cells produce eNOS mRNA and protein while eNOS polypeptide was undetectable in JEG-3 cells. In BeWo cells addition of both cytokines decreases eNOS mRNA and protein abundancies within 24 h of incubation while each substance alone had no effect. Compared to controls, the amount of eNOS transcripts was found to be elevated at low-oxygen tension, however, cNOS protein was downregulated after 24 h in the hypoxic environment, as shown by immunocytochemistry and Western blot analysis. Forskolin and methotrexate, which induce biochemical differentiation/ growth arrest in choriocarcinoma cells, stimulate eNOS mRNA and protein synthesis, but cannot overcome the decline of eNOS polypeptide levels during hypoxic incubation. It is speculated that acute hypoxia and inflammation impair eNOS/NO production of the trophoblast in vivo, which might contribute to pathological conditions of gestational diseases.

Inhibitory effect of pentoxifylline on HLA-DR expression and glycosaminoglycan synthesis by retrobulbar fibroblasts.

OBJECTIVE: Glycosaminoglycan (GAG) production by retro-ocular fibroblasts (REF) is increased in patients with thyroid-associated ophthalmopathy (TAO). Various cytokines stimulate REFs to proliferate and elaborate GAG, free oxygen radicals as well as induce HLA-DR expression on these cells. Pentoxifyllin (Ptx) regulates the production of several cytokines including tumor necrosis factor alpha (TNF-alpha), interleukin-1 (IL-1) and, interferon gamma (IFN-gamma). We wished in this study to determine whether Ptx modified the spontaneous and cytokine-induced GAG synthesis by REF and IFN-gamma induced HLA-DR expression. DESIGN: REF derived from extraocular muscles of healthy subjects were cultured without and with cytokines (IFN-gamma, TNF alpha and IL-1) and the effect of Ptx on the production of GAG by REF and HLA-DR expression was determined. MEASUREMENTS: Glycosaminoglycan was measured by incorporation of (3H) glycosamine into GAG. HLA-DR expression was analyzed by fluorescence activated cell sorter. RESULTS: Both spontaneous and cytokine induced GAG synthesis by REF was inhibited by Ptx (100, 500 and 1000 mg/l, respectively). IFN-gamma (50, 100 and 500 U/ml) induced a dose-dependent increase in the expression of HLA-DR molecules by REF. Ptx, which was not toxic to REF, inhibited HLA-DR expression on those cells dose-dependently. CONCLUSIONS: Our in vitro results suggest that Ptx reduces cytokine-induced GAG production and HLA-DR expression by REF. It thus has potential as a therapeutic agent which regulates the function of lymphocytes infiltrating the retro-orbital tissues, and which are instrumental in TAO.

Lectin histochemistry of fallopian tube epithelial cells. Relation to ovum transport and ovum pickup.

OBJECTIVE: Data on histochemical and biochemical characteristics of the human oviduct are scarce. The exact mechanisms of ovum transport and pickup are not fully understood. STUDY DESIGN: Human fallopian tubes were obtained and prepared for histochemistry. We analyzed the distribution of negatively charged groups on the oviduct epithelium and cumulus cells and examined the distribution of glycoconjugates by means of lectin histochemistry. We tested the possible influence of poly-L-lysine and considered ABO blood group expression since these characteristics are determined by specific terminal sugar residues. RESULTS: A negatively charged glycocalyx exists on tubal epithelial cells and cumulus cells. Adherence by affinities similar to sugar-lectin binding forces could be disproven in case of commonly used lectins. Poly-L-lysine inhibited the cationic binding reaction but did not influence lectin binding. The blood group A glycoprotein presents terminal D-N-acetyl-galactosamine residues, which are demonstrated by HPA lectin binding. CONCLUSION: Our study indicates that it is unlikely that electrostatic interactions play a major role in ovum transport or pickup. Since poly-L-lysine has been described as inhibiting ovum transport, sugar-lectin binding affinities seem not to operate in ovum transport or pickup.