Biomedical subjects

Y J Lu

Publications and source records attributed to Y J Lu.

At least 19 recordsLinked to original sources

PubMed2001

Comparative expressed sequence hybridization to chromosomes for tumor classification and identification of genomic regions of differential gene expression.

Altered expression of genes can have phenotypic consequences in cancer development and treatment, developmental abnormalities, and differentiation processes. Here we describe a rapid approach, termed comparative expressed sequence hybridization (CESH), which gives a genome-wide view of relative expression patterns within tissues according to chromosomal location. No prior knowledge of genes or cloning is required, and minimal amounts of tissue can be used. Expression profiles are achieved in a manner similar to the identification of chromosomal imbalances by comparative genomic hybridization analysis. The approach is demonstrated to indicate a chromosomal region that harbors overexpressed genes that may be associated with a drug-resistant phenotype. In addition, known and new regions of differential gene expression in both normal tissues and tumor samples from the soft tissue sarcoma group of rhabdomyosarcoma (RMS) are indicated. These regions included 2p24; overexpression of MYCN at 2p24 was confirmed by quantitative reverse transcription-PCR for all of the alveolar RMS cases and did not necessarily correspond to genomic amplification. Evidence including region specific microarray analysis indicated that overexpression of several genes from a region may be required for detection by CESH. This evidence is consistent with clusters of functionally related genes and mechanisms that affect the expression of a number of genes at a particular genomic location. The distinctive CESH profiles demonstrated in different subtypes of RMS show potential for tumor classification.

Chromosomes, Artificial, Yeast↗

PubMed2001

Chromosomal imbalances associated with carcinoma in situ and associated testicular germ cell tumours of adolescents and adults.

Carcinoma in situ (CIS) or intratubular germ cell neoplasia is generally considered the precursor lesion of adult testicular germ cell tumours (TGCT). The chromosomal imbalances associated with CIS and the corresponding seminoma (SE) or nonseminoma (NS) have been determined by comparative genomic hybridization (CGH) analysis of microdissected material from seven cases. Significantly, the CIS showed no gain of 12p material whereas in the invasive components of all cases gain of 12p was found, in 2 cases associated with amplification of the 12p11.2-12.1 region. Interphase fluorescence in situ analysis was consistent with this and provided evidence for the i(12p) or 12p11.2-12.1 amplification in the SE and NS but not in the corresponding CIS. This suggests a role for these changes in progression of CIS to invasive testicular cancer or progression of the invasive disease. Other imbalances such as gain of material from chromosomes 1, 5, 7, 8, 12q and X and loss of material from chromosome 18 were frequently identified (> 40% of cases) in the CIS associated with both SE and NS as well as in the invasive components. Loss of material from chromosome 4 and 13 and gain of 2p were more frequently found in the invasive components. The results shed light on the genetic relationship between the non-invasive and invasive components of testicular cancer and the stage at which particular chromosomal changes may be important.

Adolescent↗

PubMed2001

Total syntheses of bengamides B and E.

Total syntheses of the cytotoxic marine natural products bengamides B and E are described. Both bengamides are prepared via amide coupling of a protected polyhydroxylated lactone intermediate 9 with a suitably substituted aminocaprolactam intermediate. Lactone 9 is prepared in five steps from commercially available alpha-D-glucoheptonic gamma-lactone. The key reactions are a selective deprotection of a 1,2-acetonide in the presence of a 1,3-acetonide and an (E)-selective olefination of an unstable aldehyde using a gem-dichromium reagent. The bengamide B lactam intermediate 10 is prepared in seven steps from commercially available (5R)-5-hydroxy-L-lysine (12). The desired S-configuration at the gamma-OH lactam position is established using the Mitsunobu reaction.

Animals↗

PubMed2001

Characterization of chromosome aberrations associated with soft-tissue leiomyosarcomas by twenty-four-color karyotyping and comparative genomic hybridization analysis.

Data on the chromosome aberrations associated with leiomyosarcomas of soft tissues are limited, complex, and incomplete. The aim of this study was to characterize genetic aberrations associated with this tumor group, to identify consistent regions of involvement and to determine correlations with clinical outcome. Chromosomes were prepared from 10 primary soft-tissue leiomyosarcoma samples, and preparations from four of them, plus the cell line SK-LMS-1, were suitable for analysis using 24-color karyotyping by multifluor fluorescence in situ hybridization. This method allowed rearranged chromosomes to be characterized, which would not have been possible by banding analysis alone. The remaining six chromosome preparations were analyzed using standard Giemsa banding. The chromosome imbalances associated with all the samples were determined by comparative genomic hybridization analysis. Taken together, the results show both intra- and intertumor heterogeneity and considerable complexity. Although no highly consistent rearrangements were found, some regions of the genome frequently were involved, including 1q21, 5p14-pter, and 20q13, which likely harbor genes that play a role in the pathogenesis of soft-tissue leiomyosarcomas. There were no obvious correlations between the chromosomal changes identified and available clinical details.

Adult↗

PubMed2001

The kinetics and thermodynamics of surfactants in solvent sublation.

Solvent sublation has been performed on very dilute solutions of one cationic surfactant, hexadecylpyridinium chloride (HPC), and one anionic surfactant, dodecylbenzenesulfonic acid (LBS). Some thermodynamic values were obtained, e.g. molecular areas, A0, which are 50.0 and 47.7 A2/molecule, respectively, for HPC and LBS, and free adsorption energies, delta G(o)ads, which are -33.17 and -43.58 kJ mol(-1), respectively, for HPC and LBS. The kinetics were determined for a range of temperatures and gas flow-rates. Although the processes of solvent sublation of the two surfactants obey first-order kinetics, the respective adsorption mechanisms of HPC and LBS in the solvent sublation process were different. The pH and the presence of KCl and ethanol had no effect on the solvent sublation of LBS. The apparent active energy was calculated as 8.11 kJ mol(-1).

Journal Article↗

PubMed2001

Gain of 1q is associated with adverse outcome in favorable histology Wilms' tumors.

Although several genes/genetic loci involved in the etiology of Wilms' tumor have been identified, little is known of the molecular changes associated with relapse. We therefore undertook an analysis by comparative genomic hybridization (CGH) of 58 tumor samples of favorable histology Wilms' tumor taken at initial diagnosis and/or relapse. Tumors with anaplastic histology were excluded as this is known to be associated with p53 mutation and a poor prognosis. A control group of 21 Wilms' tumors that did not relapse was also analyzed. The overall frequency of gains or losses of genetic material detected by CGH was similar in both groups (77% in relapsing tumors and 70% in the nonrelapse group) as was the median number of changes per tumor (relapse group: n = 4, range, 1 to 19; nonrelapse group: n = 3, range, 1 to 8). However, gain of 1q was significantly more frequent in the relapse series [27 of 46 (59%) versus 5 of 21 (24%), P: = 0.019]. In 12 matched tumor pairs, the CGH profiles, including 1q gain, were similar at diagnosis and relapse, with little evidence for further copy number changes being involved in clonal evolution. The results suggest that 1q gain at diagnosis could be used to identify patients with favorable histology Wilms' tumor at increased risk of relapse who might benefit from early treatment intensification.

Chromosome Aberrations↗

PubMed2001

Pinworm control and risk factors of pinworm infection among primary-school children in Taiwan.

Longitudinal pinworm (Enterobius vermicularis) infection rates were estimated at a mass screening for first-grade children during 1991-1996; children were provided medication at the screening. This campaign was able to decrease the infection rates for the 1991 cohort from 16.3% to 0.6%. A case-control study was further conducted for the investigation of risk factors among fourth-graders. Cases comprised 429 children with at least one infection between September 1996 to June 1999, and controls were 280 randomly selected uninfected classmates. Parents were asked to complete a questionnaire survey to report students' personnel hygiene habits. The case-control study revealed that significant factors associated with the infection included playing on the floor (odds ratio [OR], 2.5), nail biting (OR, 2.1), failure to wash hands before meals (OR, 1.7) and living in nonapartment dwellings (OR, 1.6). Girls were at a higher, but not significant, risk (OR, 1.4), than boys. In conclusion, inadequate personal hygiene increases the risk for pinworm infection. The mass screening-medication campaign can be adapted to countries with a similar parasitic problem.

Case-Control Studies↗

PubMed2000

Trimeric ring-like structure of ArsA ATPase.

ArsA protein is the soluble subunit of the Ars anion pump in the Escherichia coli membrane which extrudes arsenite or antimonite from the cytoplasm. The molecular weight of the subunit is 63 kDa. In the cell it hydrolyzes ATP, and the energy released is used by the membrane-bound subunit ArsB to transport the substrates across the membrane. We have obtained two-dimensional crystals of ArsA in the presence of arsenite on negatively-charged lipid monolayer composed of DMPS and DOPC. These crystals have been studied using electron microscopy of negatively-stained specimens followed by image processing. The projection map obtained at 2.4 nm resolution reveals a ring-like structure with threefold symmetry. Many molecular assemblies with the same ring-shape and dimensions were also seen dispersed on electron microscopy grids, prepared directly from purified ArsA protein solution. Size-exclusion chromatography of the protein sample with arsenite present revealed that the majority of the protein particles in solution have a molecular weight of about 180 kDa. Based on these experiments, we conclude that in solution the ArsA ATPase with substrate bound is mainly in a trimeric form.

Adenosine Triphosphatases↗

PubMed2000

The expression of DAZL1 in the ovary of the human female fetus.

OBJECTIVE: To determine whether DAZL1 is expressed in human fetal ovarian tissue. DESIGN: The presence of DAZL1 expression was determined by reverse transcriptase polymerase chain reaction (RT-PCR). SETTING: Academic tertiary care medical center and research unit of university. PATIENT(S): Five female abortuses between the 19th and 22nd week of gestational age. INTERVENTION(S): Fetal ovarian tissues were collected immediately after the cessation of the heart beat. MAIN OUTCOME MEASURE(S): The product of RT-PCR. RESULT(S): DAZL1 expression was detected in all five samples. CONCLUSION(S): DAZL1 is not only expressed in human testes but also in ovaries. It may play a role in germ cell survival and gonad development in both sexes.

Abortion, Induced↗

PubMed2000

Evaluation of 24-color multifluor-fluorescence in-situ hybridization (M-FISH) karyotyping by comparison with reverse chromosome painting of the human breast cancer cell line T-47D.

Multifluor-fluorescence in-situ hybridization (M-FISH) chromosome paints for all the chromosomes in the human complement labeled with different combinations of fluorochromes is a recent technological development enabling assignment of chromosomal material to rearranged chromosomes. Little data is available on the accuracy and limitations of the approach to the analysis of complex karyotypes, which are characteristic of many malignant diseases. Here we compare M-FISH analysis of the breast-cancer-derived cell line T-47D with a previous analysis by reverse chromosome painting analysis of flow-sorted chromosomes from the same material. This demonstrated a high degree of concordance. It also illustrated the limitations of M-FISH analysis, including difficulties identifying small regions of chromosomal material and intrachromosomal rearrangements. Confirmation of selected aberrations using less-complex mixtures of painting probes and further definition of abnormalities using single copy markers may be required. The detailed karyotype description possible by M-FISH analysis contrasts with the definition in the original G-banding analysis. This and the level of concordance with reverse FISH painting supports the utility of the approach in the definition of complex karyotypes.

Breast Neoplasms↗

PubMed2000

Two different timings of intrauterine insemination for non-male infertility.

PURPOSE: Our purpose was to assess the simplicity and convenience of treatment scheduled not on weekends, by comparing two different timings of intrauterine insemination (IUI) protocol. METHODS: A prospective observational study of two different protocols of intrauterine insemination was designed. Two hundred and ten infertile couples with normal spermiograms were included in this study. Fifty-eight couples were treated with IUI 26 to 28 h after human chorionic gonadotropin (hCG) injection plus timed intercourse within a 12- to 18-hr period and 147 couples had IUI 36 to 38 hr after hCG injection and timed intercourse within a 12- to 18-hour period. Pregnancy rates were compared with two different protocols of IUI. RESULTS: The mean age, duration, and causes of infertility and the cycle characteristics following follicular stimulation were similar between the two groups. The cycle characteristics of follicular stimulation in the two treatment groups were not different. There also were no significant differences between the groups in the type of sperm concentration, sperm motility, and the percentage of sperm with normal morphology per insemination. The number of follicles greater than 17 mm per patient was not significantly different between the two groups. The pregnancy rate per cycle also was similar between the two groups in men with lower motile sperm numbers (< 40 x 10(6)) (23.6% vs. 23.4%) and in men with higher sperm numbers (> or = 40 x 10(6)) (25% vs. 24.4%). CONCLUSIONS: The different timing but similar efficacy of these two IUI protocols provides a practical choice to clinicians. The availability of both protocols may avoid unnecessary scheduling of clinical and laboratory work on weekends and holidays in women participating in controlled ovarian hyperstimulation and IUI programs for treatment of non-male infertility.

Adult↗

PubMed2000

Comparative genomic hybridization analysis of myoepithelial carcinoma of the breast.

Although there seems to be a common stem cell for the two epithelial cell types in the breast, the vast majority of breast cancers exhibit a luminal phenotype. Pure myoepithelial carcinomas are rare. We report our findings of genetic alterations in these tumors. We have analyzed 10 cases of pure myoepithelial cell carcinomas using laser capture microdissection and comparative genomic hybridization. The mean number of changes was 2.1 (range 0-4), compared with a mean of 8.6 (range 3.6-13.8) in unselected ductal carcinomas. Common alterations included loss at 16q (3/10 cases), 17p (3/10), 11q (2/10), and 16p (2/10), regions also commonly deleted in ductal carcinomas. The single case in which both pure myoepithelial carcinoma and invasive ductal carcinoma was present showed 2 alterations in the myoepithelial tumor (losses at 17p and 17q), whereas the invasive ductal component showed 15 alterations (5 gains and 9 losses), including loss at 17p. The sharing of 17p loss in myoepithelial and ductal carcinoma is consistent with a common stem cell model in the breast. The relatively few genetic alterations in otherwise aggressive neoplasms suggests that myoepithelial tumors may be a good model for the delineation of genes important in breast tumorigenesis.

Aged↗

PubMed2000

Clinical implications of intracytoplasmic sperm injection using cryopreserved testicular spermatozoa from men with azoospermia.

OBJECTIVE: To investigate whether sperm obtained by testicular sperm extraction (TESE) and cryopreserved well before intracytoplasmic sperm injection (ICSI) can serve as an effective sperm source. STUDY DESIGN: The role of cryopreserved testicular spermatozoa was evaluated in a retrospective analysis of consecutive ICSI cycles using fresh or cryopreserved sperm; they were followed by prospective, planned treatment using cryopreserved sperm with a modified ICSI procedure. Sixteen men (22 cycles) with obstructive or nonobstructive azoospermia were included in the retrospective analysis. Another 25 men (29 cycles) were in the planned treatment group. Following these series, the pregnancy outcomes were compared between ICSI cycles with fresh or cryopreserved testicular sperm. RESULTS: In the retrospective phase, 14 ICSI cycles were performed using fresh sperm, with 8 using cryopreserved sperm. There were no statistically significant differences between the two groups in any outcome measure. Planned treatment with cryopreserved sperm resulted in a fertilization rate of 84% and an embryo transfer rate of 89%. Thirteen couples (44%) achieved pregnancy (five ongoing, six delivered). These rates were similar to those in the retrospective phase of the study. All couples in the planned cryopreservation group had multiple aliquots (6.5 +/- 2.1) of sperm remaining after the first cycle. CONCLUSION: Cryopreserved sperm obtained by TESE can be used as an effective sperm source in ICSI cycles. Planned cryopreservation allows multiple aliquots to be stored for use in subsequent cycles and thus avoids the need for repeat biopsies.

Adult↗

PubMed1999

Cloning and mapping of members of the MYM family.

Tandem repeats of a novel, putative, zinc-binding motif (MYM) have been described within the products of two, highly homologous genes: ZNF198/RAMP/FIM and ZNF261/DXS6673E. ZNF198, mapping to 13q11-q12, was recently shown to fuse to the fibroblast growth factor receptor 1 gene in the t(8;13)(p11;q11-q12) rearrangement associated with a stem cell leukemia/lymphoma syndrome. ZNF261 at Xq13.1 is disrupted by a t(X;13)(q13.1;q32) rearrangement in a mentally retarded patient and is a candidate gene for nonspecific X-linked mental retardation. Here we have cloned another member of this family, designated ZNF258, and mapped it to chromosome band 14q12. In addition, ZNF262/KIAA0425 was identified as a further member of the family and mapped to 1p32-p34. The predicted protein products of ZNF258 and ZNF262 maintain the repeats of the MYM motif. Isolation of these new members will facilitate the functional characterization of the MYM family and motif.

Amino Acid Motifs↗

PubMed1999

Synovial sarcoma specific translocation associated with both epithelial and spindle cell components.

The biphasic subtype of synovial sarcoma consists of both epithelial and spindle cell components. To address the relationship between the different cellular components found in synovial sarcoma, we deduced the presence of the synovial sarcoma-specific der(X)t(X;18)(p11.2;q11.2), and involvement of the SSX1 gene in both the epithelial/glandular and spindle cell components of 3 biphasic synovial sarcomas with wide ranging proportions of each of the 2 elements. This has been achieved using a 2-color fluorescence in situ hybridization (FISH) methodology that we had developed recently for analysis paraffin-embedded material. The presence of the rearrangement could be deduced in histologically defined regions and the results showed clearly that the rearrangement was present in both cellular components. This finding provides direct genetic evidence for biphasic synovial sarcomas being clonal and truly biphasic.

Adult↗

PubMed1999

A comparative molecular field analysis study on several bioactive peptides using the alignment rules derived from identification of commonly exposed groups.

A 3D convex hull computation algorithm designed by us previously is used as a tool to align a series of structures randomly generated for eleven bioactive tachykinin peptides. There are 10 random structures generated for each peptide. A random structure is selected for each peptide to form a structural set of eleven structures. The total number of structural sets generated is 100. The convex hull computation algorithm is applied to each peptide structure generated. We count the frequency of atoms lying on the vertices of each hull computed. Vertices of the same atom type are gathered together as a set of commonly exposed atoms for a structural set generated. Structures are then aligned by treating the set of commonly exposed atoms as a set of correspondences using the FIT option of the SYBYL 6.4 program. All the structure sets are also aligned by using the coordinates of the backbone C alpha atoms as a set of correspondences through the same program. It is found that while a smaller degree of structural similarity for structures aligned by the convex hull alignment rule is detected, the overall SYBYL comparative molecular field analysis (CoMFA) statistics computed for the aligned structures using the alignment rule is better than that computed for the aligned structures using the C alpha atoms alignment rule. A similar conclusion is drawn for a subset of structures selected and probed by a different type of atoms using the SYBYL CoMFA program. These results indicate that computation of 3D convex hulls is a feasible way that one can use to align structures generated for highly flexible molecules of this kind.

Algorithms↗

PubMed1999

Dual colour fluorescence in situ hybridization to paraffin-embedded samples to deduce the presence of the der(X)t(X;18)(p11.2;q11.2) and involvement of either the SSX1 or SSX2 gene: a diagnostic and prognostic aid for synovial sarcoma.

Identification of the t(X;18)(p11.2;q11.2) and the fusion gene products, SYT-SSX1 and SYT-SSX2, associated with a high proportion of synovial sarcomas, has been shown to be a useful diagnostic aid. This study demonstrates the application of dual colour fluorescence in situ hybridization to paraffin-embedded samples to deduce the presence of the derivative X chromosome and also the position of the breakpoint on chromosome X at either the SSX1 or the SSX2 gene. This used region specific markers from chromosomes X and 18 and an optimized protocol involving microwave exposure. Novel and rapid scoring criteria were validated which circumvented potential problems of nuclear truncation and defining cell boundaries. This involved blind analysis of two negative sarcoma samples and three synovial sarcomas in which corresponding frozen material had been previously shown to have the translocation involving different SSX genes. Six new cases diagnosed as synovial sarcoma were also analysed; two monophasic and two biphasic case were deduced to have a breakpoint in the SSX1 gene, one monophasic case an SSX2 breakpoint, and one case did not show rearrangement of the region. The ability to analyse formalin-fixed, paraffin-embedded samples in this way has practical implications for aiding the diagnosis of difficult cases, recently ascribed prognostic relevance, and allows further retrospective studies to be carried out. The methodology is also applicable to the identification of other tumour specific translocations in paraffin-embedded material.

Adult↗

PubMed1999

Chromosome 3 imbalances are the most frequent aberration found in non-small cell lung carcinoma.

The chromosomal imbalances in nine cases of primary non-small cell lung cancer (NSCLC) and two cell lines derived from normal human bronchial epithelial (HBE) tissue were identified by comparative genomic hybridization (CGH). Gain of material from 3q and loss of 3p material were the most frequent changes in the primary tumors. Other commonly found imbalances included gain of material from 1q, 7p, 8q, 9q, 17q and 20q, and losses involving 4, 5q, 8p, 10 and 13q. High level gain was found in two cases, both encompassing the 3q23-q27 region. Loss of 3p was also found in both of the HBE cell lines suggesting that loss of one or more tumor supressor genes on 3p may be important for epithelial transformation and could be involved in the earlier stages of lung cancer development.

Adult↗