Highly Sensitive, Robust, and Recyclable TiO2/AgNP Substrate for SERS Detection.

Label-free biosensors provide an important platform for detecting chemical and biological substances without needing extra labeling agents. Unlike surface-based techniques such as surface plasmon resonance (SPR), interference, and ellipsometry, surface-enhanced Raman spectroscopy (SERS) possesses the advantage of monitoring analytes both on surfaces and in solutions. Increasing the SERS enhancement is crucial to preparing high-quality substrates without quickly losing their stability, sensitivity, and repeatability. However, fabrication methods based on wet chemistry, nanoimprint lithography, spark discharge, and laser ablation have drawbacks of waste of time, complicated processes, or nonreproducibility in surface topography. This study reports the preparation of recyclable TiO2/Ag nanoparticle (AgNP) substrates by using simple arc ion plating and direct-current (dc) magnetron sputtering technologies. The deposited anatase-phased TiO2 ensured the photocatalytic degradation of analytes. By measuring the Raman spectra of rhodamine 6G (R6G) in titrated concentrations, a limit of detection (LOD) of 10-8 M and a SERS enhancement factor (EF) of 1.01 × 109 were attained. Self-cleaning was performed via UV irradiation, and recyclability was achieved after at least five cycles of detection and degradation. The proposed TiO2/AgNP substrates have the potential to serve as eco-friendly SERS enhancers for label-free detection of various chemical and biological substances.

Comparative modeling of improved synthesis of energetic dinitrobenzofuroxan (DNBF) derivatives.

Quantum chemical theoretical computation was performed on gaseous molecular reaction systems to simulate parallel synthesis of energetic primary explosive precursor 4,6-dinitrobenzofuroxan (4,6-DNBF) and its isomeric derivatives. Related polarized continuum model (PCM) and Materials Studio (MS/forcite) energies were collected via kinetic rate and thermodynamic equilibrium analyses, enabling comparison of and suggestions as to suitable reaction conditions (reaction temperature, reagent concentration, mixed acid ratio) together with feasible pathways to obtain a high production yield of the research target. In summary, at a low reaction temperature of 278 K, 1.0 M 4-nitrobenzofuroxan (or 5,6-nitrobenzofuroxan) could be nitrated using concentrated nitric acid/sulfuric acid at a 1 to 2 volume ratio to efficiently and rapidly produce 4,6-dinitro-benzofuroxan (or 5,6-dinitrobenzofuroxan), in agreement with the experimental results reported in the literature. Graphical abstract.

Prerequisite OCT4 Maintenance Potentiates the Neural Induction of Differentiating Human Embryonic Stem Cells and Induced Pluripotent Stem Cells.

Establishing an efficient differentiation procedure is prerequisite for the cell transplantation of pluripotent stem cells. Activating fibroblast growth factor (FGF) signals and inhibiting the activin/nodal pathway are both conserved principles to direct the neural induction (NI) of developing embryos and human embryonic stem cells (hESCs). Wnt signal and OCT4 expression are critical for the hESC pluripotency; however, their roles in cell differentiation are largely unclear. We demonstrate that in the presence of FGF2 and activin inhibitor SB431542, applying a small-molecule Wnt agonist, BIO, efficiently and rapidly steers the NI of all our tested hESCs. A human induced pluripotent stem cell (iPSC), which is refractory for efficient neural conversion by FGF2, effectively differentiated to SOX1(+) cells after the BIO/SB431542/FGF2 treatment. In addition, BIO promoted cell survival and transiently sustained OCT4 expression at the early NI stage with FGF2 and SB431542. Interestingly, at the late NI stage, the OCT4 level rapidly declined in the treated hESCs and consequently initiated the formation of neural rosettes with forebrain neuron characteristics. This study illustrates the distinct effects of Wnt activation on maintaining pluripotency and committing neural lineages at the early and late NI stages of hESCs and iPSCs, respectively.

Stereotactic ablative radiotherapy: a potentially curable approach to early stage multiple primary lung cancer.

BACKGROUND: Surgical resection has been the standard treatment for early stage multiple primary lung cancer (MPLC). However, a significant proportion of patients with MPLC cannot undergo surgery. For this report, the authors explored the role of stereotactic ablative radiotherapy (SABR) for patients with MPLC. METHODS: Patients with MPLC who received SABR (50 grays [Gy] in 4 fractions or 70 Gy in 10 fractions) for the second tumor were reviewed. Four-dimensional, computed tomography-based, planning/volumetric image-guided treatment was used for all patients. Treatment outcomes/toxicities were analyzed. RESULTS: For the 101 patients who received SABR, at a median follow-up of 36 months and with a median overall survival (OS) of 46 months, the 2-year and 4-year in-field local control rates were 97.4% and 95.7%, respectively. The 2-year and 4-year OS rates were 73.2% and 47.5%, respectively; and the progression-free survival (PFS) rates were 67% and 58%, respectively. Patients who had metachronous tumors had better OS and PFS than patients who had synchronous tumors (2-year OS: 80.6% metachronous vs 61.5% synchronous; 4-year OS: 52.7% vs 39.7%, respectively; P = .047; 2-year PFS: 84.7% vs 49.4%, respectively; 4-year PFS: 75.6% vs 30.4%, respectively; P = .0001). For patients who either underwent surgery or received SABR for an index tumor, the incidence of grade ≥ 3 radiation pneumonitis was 3% (2 of 71 patients); however, this increased to 17% (5 of 30 patients) for those who received conventional radiotherapy for an index tumor. Other grade ≥ 3 toxicities included grade 3 chest wall pain (3 of 101 patients; 3%) and grade 3 skin toxicity (1 of 101 patients; 1%). CONCLUSIONS: SABR achieves promising long-term tumor control and survival and may be a potential curative treatment for early stage MPLC.

Target identification of microRNAs expressed highly in human embryonic stem cells.

MicroRNAs (miRNAs) are noncoding RNAs of approximately 22 nucleotides in length that negatively regulate the post-transcriptional expression by translational repression and/or destabilization of protein-coding mRNAs. The impact of miRNAs on protein output was recently shown that although some targets were repressed without detectable changes in mRNA levels, those translationally repressed by more than a third also displayed detectable mRNA destabilization, and, for the more highly repressed targets, mRNA destabilization usually comprised the major component of repression. Thus, comparative profilings of miRNAs and mRNAs from the same samples of different cell types may identify the putative targets of miRNAs. In this investigation, both miRNA and mRNA profiles from the undifferentiated human embryonic stem cell line hES-T3 (T3ES), hES-T3 derived embryoid bodies (T3EB), and hES-T3 differentiated fibroblast-like cells (T3DF) were compared, and 58 genes were found to be targets of four hES cell-specific miRNAs miR-302d, miR-372, miR-200c and/or miR-367 by inverse expression levels (highly negative correlation) of miRNAs to their target mRNAs. Approximately half of these 58 targets are involved in gene transcription. Three common target genes TRPS1, KLF13 and MBNL2 of three highly expressed miRNAs miR-302d, miR-372, and miR-200c were identified, and the target sites of both miR-302d and miR-372 in the 3'UTR of TRPS1, KLF13, and MBNL2 genes were confirmed by the luciferase assay. The highly expressed mRNAs and miRNA target mRNAs involved in KEGG pathways among T3ES, T3EB, and T3DF cells were also compared, and the expression levels of target mRNAs predicted by abundantly expressed miRNAs were found to be three- to sixfold lower than those of non-target mRNAs involved in the same signaling pathways.

Serum resistin level is a predictor of ovarian response in in vitro fertilisation cycle.

OBJECTIVES: To measure serum resistin levels in infertile women undergoing in vitro fertilisation-embryo transfer (IVF-ET), and to find any correlations between serum resistin levels and body weight, body mass index, the number of oocytes retrieved, and the outcome of IVF-ET. In addition, to assess whether there is any difference in serum resistin levels between infertility caused by polycystic ovary syndrome (PCOS) and infertility caused by other female factors. METHODS: We designed a case-control study, and a total of 44 infertile women were enrolled. The blood samples for resistin measurement were collected on day 3 of the menstrual cycle prior to the administration of gonadotropin during in vitro fertilisation. These cases were then divided into 2 subgroups (PCOS group versus non-PCOS group) and a number of variables were measured and compared, including serum resistin levels. RESULTS: Serum resistin levels were inversely correlated with the number of oocytes retrieved (r=-0.371, p=0.013). No significant correlation was found between serum resistin levels and body mass index or body weight, either in the whole group or in the 2 subgroups. Serum resistin levels in the non-PCOS group were significantly higher than in the PCOS group (p=0.049). Serum resistin levels in the non-PCOS group were inversely correlated to the number of oocytes retrieved (r=-0.386, p=0.039), but no similar correlation was found in the PCOS group. There was no correlation between serum resistin levels and fertility rate or clinical pregnancy rate in either subgroup. CONCLUSIONS: We observed a negative correlation between serum resistin levels and the number of oocytes retrieved during IVF. However, this phenomenon was only present in the non-PCOS group. This result suggests that serum resistin levels might be a good predictor of ovarian response in infertile women without PCOS during IVF. The role of serum resistin in response to inflammation caused by endometriosis or chronic pelvic infection, both of which are major causes of female infertility, should be examined in a further study.

Characterization and gene expression profiling of five new human embryonic stem cell lines derived in Taiwan.

Many human embryonic stem cell (hESC) lines have been reported, but only a few of them have been fully characterized. In this report, five new hESC lines were derived from 32 discarded blastocysts in Taiwan, and these lines were continuously cultured on mitotically inactivated mouse embryonic fibroblast (MEF) feeder layer in the hESC medium for more than 44 passages and underwent freezing/thawing processes. All five hESC lines expressed characteristic undifferentiated hESC markers, such as SSEA-4, TRA-1-81, alkaline phosphatase, TERT, and the transcription factors POU5F1 (OCT4) and NANOG. hESC lines T1 and T3 possess normal female karyotypes, whereas lines T4 and T5 are normal male, but line T2 is male trisomy 12 (47XY,+12). hESC lines T1, T2, T3, and T5 were able to produce teratomas in severe combined immunodeficient (SCID) mice, and line T4 could only form embryoid bodies (EBs) in vitro. Global gene expression profiles of these five newly derived hESC lines were analyzed using the Affymetrix human genome U133 plus 2.0 GeneChip. The results showed that 4,145 transcripts, including 19% of unknown functions, were detected in all five hESC lines. Comparison of the 4,145 genes commonly expressed in the five hESC lines with those genes expressed in teratomas produced by the hESC line T1 and placenta revealed 40 genes exclusively expressed in all five hESC lines. These 40 genes include the previously reported stemness genes, such as POU5F1 (OCT4), NANOG, TDGF1 (CRIPTO), SALL4, LECT1, and BUB1 responsible for self-renewal and pluripotent differentiation. The global gene expression analysis also indicated that the transforming growth factor-beta (TGF-beta)/activin branch components inhibin BC, ACVR2A, ACVR1 (ALK2), TGFBR1 (ALK5), and SMAD2 were found to be highly expressed in undifferentiated states of these five hESC lines and decreased upon differentiation. In short, the hESC nature of these five hESC lines is supported by the undifferentiated state, extensive renewal capacity, and pluripotency, including the ability to form teratomas and/or EBs. These cell lines will be useful for human embryonic stem cell biology and drug development.

Serum adiponectin levels increase after human chorionic gonadotropin treatment during in vitro fertilization.

AIMS: To determine whether or not serum adiponectin concentrations are influenced by ovarian hyperstimulation during in vitro fertilization (IVF). METHODS: This study involved 52 women who were participating in IVF-ET cycles. Adiponectin levels in serum were determined by radioimmunoassay and compared. RESULTS: Serum adiponectin levels fell from Day-basal to Day-hCG (p = 0.047), and then rose on Day-OR and again on Day-7ET (p < 0.001; p < 0.001). Estradiol levels on Day-hCG were significantly and positively correlated with serum adiponectin levels on Day-OR and Day-7ET (r = 0.325, p = 0.019; r = 0.372, p = 0.007). Progesterone levels on Day-OR positively correlated with serum adiponectin levels on Day-basal (r = 0.278, p = 0.046). There was also a positive correlation between progesterone levels on Day-7ET and serum adiponectin levels on Day-OR (r = 0.289, p = 0.038). Multiple linear regression analysis revealed that adiponectin levels on Day-OR and Day-7ET were negatively correlated with age and body mass index after adjustment was made for concomitant diseases. CONCLUSIONS: To sum up, following gonadotropin treatment, serum adiponectin levels decrease as a result of the negative effect of high estradiol levels on adiponectin production. Conversely, serum adiponectin levels increase following human chorionic gonadotropin treatment.

Analysis of gene expression in single human oocytes and preimplantation embryos.

Little is known about the gene expression in human oocytes and early embryo development because of the rare availability of the materials. The recent advancement of biotechnology has allowed one to analyze the gene expression in single human oocytes and preimplantation embryos. Gene expression of human lactate dehydrogenase isozymes (LDH-A, LDH-B, and LDH-C) and small ubiquitin-like modifier isoforms (SUMO-1, SUMO-2, and SUMO-3) in four oocytes, two 4-cell and three 8-cell embryos was studied using the reverse transcription-polymerase chain reaction. The mRNAs for SUMO-1, SUMO-2, SUMO-3, and LDH-B (heart) were detected in all of oocytes, 4- and 8-cell embryos. The mRNA for LDH-A (muscle) was detected in two of four oocytes and one of three 8-cell embryos. However, the mRNA for testis-specific LDH-C was not detected at all as expected. A cDNA microarray containing 9600 cDNA spots was used to investigate differential expression of human genes in oocyte, 4-cell and 8-cell embryos. The expression of 184, 29, and 65 genes was found to have a value more than twofold above the median value of all genes expressed in oocyte, 4- and 8-cell embryos, respectively, indicating that the expression of some zygotic genes had already occurred at 4-cell embryo.

Higher basal adiponectin levels are associated with better ovarian response to gonadotropin stimulation during in vitro fertilization.

AIMS: To measure adiponectin levels in the serum samples from day 3 of the menstrual cycle prior to the administration of gonadotropin during in vitro fertilization, and to measure any correlations between adiponectin levels and the number of oocytes retrieved, body weights and body mass indexes. Also, to examine whether there is a difference in adiponectin levels between women who conceive and those who do not. METHODS: In a case-control design, 56 women undergoing in vitro fertilization procedures were included in this study: 28 women who conceived were matched with 28 women who did not conceive. Adiponectin levels in serum were determined by radioimmunoassay and compared. RESULTS: The adiponectin levels were positively correlated with the number of oocytes retrieved (r = 0.306, p = 0.022), but negatively correlated with body mass index and body weight (r = -0.367, p = 0.005; r = -0.326, p = 0.014). No significant correlations were found between the number of oocytes retrieved and body mass index or body weight (r = 0.020, p = 0.882; r = 0.069, p = 0.613). We further observed that adiponectin levels in women who conceived (23.0 +/- 2.0 microg/ml) were found to be significantly higher (p = 0.026) than those in women who did not conceive (17.3 +/-1.4 microg/ml). The number of oocytes retrieved from women who conceived (13.3 +/- 0.9) was also higher than that in women who did not (10 +/- 1.2; p = 0.029). CONCLUSIONS: The number of oocytes retrieved was found to correlate positively with the adiponectin levels on the day prior to the administration of gonadotropin. In addition, basal adiponectin levels were significantly higher in those women who conceived. Therefore, adiponectin is a better marker of adequate follicular development during in vitro fertilization than is body weight or body mass index.

Introduction of a laparoscopic gynecological program in a general hospital in Taiwan.

BACKGROUND AND OBJECTIVES: We reviewed the records for 571 gynecologic laparoscopies performed at a privately owned general hospital in Kaosiung Taiwan in 1998 and 1999 and discuss here the major obstacles we encountered while introducing these procedures at our institution. METHODS: Included in this series are 293 procedures performed in 1998 (149 hysterectomies, 144 adnexal procedures), and 278 procedures performed in 1999 (131 hysterectomies, 147 adnexal procedures). Thirty-nine of these patients also underwent laparoscopic appendectomy. Mean patient age was 62 years (range 28 to 82). All procedures were performed by 1 of 6 board-certified gynecologic surgeons, or by 1 of 4 residents under the direct supervision of a board-certified surgeon. RESULTS: We experienced 0% mortality and 7.2% morbidity in this series. In comparing cases from 1998 and 1999, we observed a decrease in both mean surgery time (135.4 to 123.0, P=0.032) and mean length of hospital stay (5.52 to 4.62, P=0.046) for hysterectomies and adnexal procedures combined. CONCLUSIONS: These data support ongoing efforts to incorporate gynecologic laparoscopy as an alternative to open procedures at our institution. Introduction of these procedures in privately owned hospitals in Taiwan has been limited because of the large initial investment for equipment, patient education issues, and difficulties obtaining reimbursement.

Laparoscopic myomectomy technique. Use of colpotomy and the harmonic scalpel.

OBJECTIVE: To describe the use of colpotomy and the harmonic scalpel in laparoscopic myomectomy and to compare estimated blood loss when using unipolar cautery versus the harmonic scalpel and surgical time for colpotomy versus morcellation. STUDY DESIGN: A retrospective, cohort study was performed on 168 patients who were diagnosed with and treated for uterine leiomyomata between January 1992 and January 2000. Patients presented with infertility, menometrorrhagia, dysmenorrhea, masses on ultrasound or a combination of these symptoms. Two patients required hysterectomy, and 1 required conversion to laparotomy due to bleeding, leaving 165 patients who underwent laparoscopic myomectomy, 143(87%) by colpotomy and 22 (13%) by morcellation. Unipolar cautery was used for uterine incision in 112 of the 165 myomectomies (68%) and the harmonic scalpel in 53 (32%). Surgery was performed at one of two community hospitals in the Seattle area or a general hospital in Taiwan. RESULTS: Mean estimated blood loss using the harmonic scalpel (243 mL, range 150-350) was significantly less (P < .01) than that using unipolar cautery (378 mL, range 203-800) for uterine incision. Mean surgical time for colpotomy (144 minutes, range 110-260) was less (P < .05) than that for morcellation (168 minutes, range 140-244) despite having removed a larger median number of myomata per patient of comparable or larger size (seven by colpotomy versus four by morcellation). CONCLUSION: These data suggest that the harmonic scalpel is effective for uterine incision during myomectomy and may result in less bleeding than unipolar cautery while offering some advantages in safety. Multiple leiomyomata can generally be extracted more quickly via posterior colpotomy than by morcellation. This difference is smaller and therefore less important in patients with only a few small to medium-sized (< 10 cm) fibroids. For these patients, the minimal invasiveness of morcellation may offer a more significant benefit.

Leptin affects pregnancy outcome of in vitro fertilization and steroidogenesis of human granulosa cells.

PURPOSE: This study was to examine the serum leptin levels in the prediction of pregnancy outcomes in women receiving ovarian hyperstimulation. Effect of leptin on the steroidogenesis was evaluated for granulosa cell (GC) culture in vitro. METHOD: Serum levels of leptin and estradiol were measured on Day 2, the time of hCG administration and oocyte retrieval in 50 women undergoing long-course GnRH agonist ovarian hyperstimulation. The production of estrogen and progesterone in granulosa cell culture were detected after administration of leptin at the absence or presence of FSH 1 mIU. RESULTS: Leptin levels at the time of hCG injection were significantly lower in the pregnant women than in those without pregnancy. Estradiol concentrations were not correlated with leptin levels during the time of hCG injection and oocyte retrieval. High leptin concentration suppressed both basal estradiol and progesterone productions in GC. Leptin in high doses suppressed estradiol production, but did not alter progesterone production of GC in the presence of FSH. CONCLUSIONS: Serum leptin levels may be predictive of IVF pregnancy outcome with the effect on intraovarian progesterone/estradiol ratio during the follicular phase. Significantly low serum leptin levels were noted in the pregnant women than in the nonpregnant women.

Laparoscopic uterine suspension by round ligament plication.

OBJECTIVE: To describe a method for laparoscopic uterine suspension by round ligament plication using standard suturing instruments. STUDY DESIGN: Forty-six women underwent uterine suspension for treatment of chronic pelvic pain due to uterine retroversion over a period of five years. All operations were performed by the principal author. RESULTS: Forty-six patients treated with this technique were selected from 388 who underwent clinical evaluation and diagnostic laparoscopy for chronic pelvic pain, defined as lasting more than six months. There were no intraoperative complications in this series. Mean follow-up was 23.6 months (range, 6-42). At each follow-up, patients were asked to rate their pain relative to preoperatively as unchanged, minor, moderate, significant or resolved. At six months, 44 of 46 patients (96%) reported at least some improvement, with 17 (37%) reporting significant improvement, and 18 (39%) reporting resolution of pain. Sixteen of 27 patients (59%) at 24 months' follow-up and 15 of 23 patients (65%) with 36 months' follow-up reported significant improvement in or resolution of their pain. CONCLUSION: For patients experiencing chronic pelvic pain associated with uterine retroversion, round ligament plication is an effective method of repositioning the uterus and reducing or eliminating the patient's symptoms.