Comparison of pregnancy outcomes following fresh and electively frozen single blastocyst transfer in natural cycle and clomiphene-stimulated IVF cycles.

STUDY QUESTION: Are there any differences in live birth rates (LBR) following fresh blastocyst transfer in natural or clomiphene-stimulated cycles, or after elective blastocyst freezing in clomiphene-stimulated cycles followed by thawing and transfer at different time-points? SUMMARY ANSWER: Clomiphene citrate (CC) administration adversely affected the LBR after single fresh blastocyst transfer (SBT) in CC cycles compared with that in natural cycles, while this adverse effect of CC is not present when a single vitrified-warmed blastocyst transfer (SVBT) is performed in subsequent natural ovulatory cycles, regardless of the duration between CC administration and the day of SVBT. WHAT IS KNOWN ALREADY: CC affects uterine receptivity associated with a thinning of the uterine endometrium through an antioestrogenic effect. However, the duration that this adverse effect of CC on uterine endometrium persists after initial use is still unknown. STUDY DESIGN SIZE DURATION: A retrospective cohort study of 157 natural cycle IVFs followed by SBT and 1496 minimal ovarian stimulation with CC IVF cycles followed by SBT (n = 24) or SVBT (n = 1472) from January 2010 to December 2014 was conducted. SVBT cycles were classified into two groups according to the period between the last day of CC administration and the day of SVBT (A: ≤60 d and B: ≥61 d). All groups were then compared based on pregnancy outcomes (natural-SBT group: n = 157, CC-SBT group: n = 24, SVBT-A: n = 1143, SVBT-B: n = 329). PARTICIPANTS/MATERIALS SETTING METHODS: Women were aged 30-39 years at oocyte retrieval. In SVBT cycles, blastocysts were vitrified and warmed using a Cryotop safety kit. SVBT was performed in subsequent natural ovulatory cycles. The main outcomes were LBR and neonatal outcome, and both were compared among the groups. MAIN RESULTS AND THE ROLE OF CHANCE: The LBR in the CC-SBT group (29.2%, 7/24) was significantly lower compared with the natural-SBT (56.1%, 88/157) (P = 0.01) and SVBT-A (50.0%, 572/1143) (P = 0.04), but not SVBT-B (47.4%, 156/329), groups. Furthermore, multivariate logistic regression analysis revealed that the LBR was comparable among the natural-SBT and SVBT groups, but was significantly lower in the CC-SBT group (adjusted odds ratio: 0.324, 95% CI: 0.119-0.800, P = 0.01). No significant differences among all groups were observed for gestational age (P = 0.19), birthweight (P = 0.41) and incidence of malformation (P = 0.53). LIMITATIONS REASONS FOR CAUTION: In this study we analysed a biased sample, based on clinical judgement regarding endometrial thickness, and the study was limited by its retrospective nature. The low statistical power caused by the group size disparity was also a limitation, especially in the CC-SBT group. Although the outcome showing inferiority of CC-SBT compared to natural-SBT is consistent with general findings in the literature, further large-scale clinical studies, ideally RCTs, are necessary to validate our results and clarify the prolonged effect of CC in SVBT cycles on pregnancy and neonatal outcomes. WIDER IMPLICATIONS OF THE FINDINGS: Our observation suggests that CC administered in minimal ovarian stimulation cycles affects adversely the pregnancy outcomes when SBT is performed. Therefore, for a CC-based minimal stimulation IVF cycle, we suggest that frozen embryo transfer should be performed in a subsequent natural ovulatory cycle to avoid the possibility of implantation failure associated with CC administration. STUDY FUNDING/COMPETING INTERESTS: The authors have no conflicts of interest to declare. No external funding was either sought or obtained.

Electrochemical microdevices for rapid and on-site determination of the minimum inhibitory concentration of antibiotics.

A tool for rapid selection of appropriate antibiotics may be useful to maximize the benefits of their effectiveness against severe infectious diseases. In this study, we propose a microdevice that works with surface tension-driven sample introduction and impedance measurements for on-site, rapid estimation of suitable antibiotics.

Glycolipid-based nanostructures with thermal-phase transition behavior functioning as solubilizers and refolding accelerators for protein aggregates.

The self-assembly of synthetic glycolipids produced nanostructures such as vesicles and nanotubes consisting of bilayer membranes, which underwent a gel-to-liquid crystalline thermal phase transition. Vesicles formed at temperatures above the thermal phase transition temperatures (Tg-l) could solubilize aggregates of denatured proteins by trapping them in the fluid bilayer membranes. Cooling to temperatures below Tg-l caused a morphological transformation into nanotubes that accompanied the thermal phase transition from the fluid to the solid state. This phenomenon allowed the trapped proteins to be quickly released into the bulk solution and simultaneously facilitated the refolding of the proteins. The refolding efficiency strongly depended on the electrostatic attraction between the bilayer membranes of the nanostructures and the proteins. Because of the long shape (>400 nm) of the nanotubes, simple membrane filtration through a pore size of 200 nm led to complete separation and recovery of the refolded proteins (3-9 nm sizes).

Three-dimensional positioning and control of colloidal objects utilizing engineered liquid crystalline defect networks.

Topological defects in liquid crystals not only affect the optical and rheological properties of the host, but can also act as scaffolds in which to trap nano or micro-sized colloidal objects. The creation of complex defect shapes, however, often involves confining the liquid crystals in curved geometries or adds complex-shaped colloidal objects, which are unsuitable for device applications. Using topologically patterned substrates, here we demonstrate the controlled generation of three-dimensional defect lines with non-trivial shapes and even chirality, in a flat slab of nematic liquid crystal. By using the defect lines as templates and the electric response of the liquid crystals, colloidal superstructures are constructed, which can be reversibly reconfigured at a voltage as low as 1.3 V. Three-dimensional engineering of the defect shapes in liquid crystals is potentially useful in the fabrication of self-healing composites and in stabilizing artificial frustrated phases.

Fabrication of patterned cell co-cultures on albumin-based substrate: applications for microfluidic devices.

A surface coated with cross-linked albumin film resists the adhesion of cells, and subsequent exposure to UV irradiation or electrostatic adsorption of a cationic polymer switches the surface from non-adherent to adherent. Taking advantage of this unique property of cross-linked albumin, the authors fabricated patterned cell co-cultures with desired patterns and cell types. In this scheme, the cell-adherent region was initially created in the cell-non-adhesive albumin substrate, on which a first cell type was attached. Subsequently, the remaining region was also changed to adherent for the attachment of secondary cells in the same manner, thereby allowing distinctly localized co-cultures. As a proof of concept demonstration of the feasibility of this approach, a patterned co-culture of Neuro-2a cells with L929 cells was successfully prepared on the substrate. Furthermore, combining this technique with a microfluidic technique, a micropatterned co-culture of PA6 cells with 3T3 fibroblasts was created inside microfluidic devices. This approach could potentially be a useful tool for fundamental investigations of cell-cell interactions and for tissue engineering applications.

Mandibular reconstruction with bone transport in a patient with osteogenesis imperfecta.

Osteogenesis imperfecta (OI) was originally considered a connective tissue disorder, primarily involving type 1 collagen. It is characterized by numerous skeletal and extraskeletal defects, including bone fragility, multiple fractures, blue sclerae, hearing deficits, skin and ligament laxity, and dentinogenesis imperfecta (DI). The authors describe a 24-year-old Japanese man with OI and DI who had an ossifying fibroma of the mandible. Segmental resection was performed, and the mandible was reconstructed by distraction osteogenesis with the transport segment technique. This is the first report to describe a patient with OI undergoing mandibular reconstruction with bone transport, to the authors' knowledge.

The BMP signaling and its Smads in mandibular distraction osteogenesis.

AIM: The aim of the present study was to clarify the mechanism of signal transduction of bone morphogenetic proteins (BMPs) through their specific down signaling molecules Smads inducing bone formation in response to mechanical stimulus during distraction osteogenesis (DO) in the rat mandible. MATERIALS AND METHODS: Osteotomy of the left mandible was performed in 45 rats. Thirty rats underwent mandibular distraction (protocol; 5 days latency, 8 days distraction, and 2 weeks consolidation) while 15 rats served as non-distracted (fracture healing) group. The expression of BMPs-2,-4 and Smads 1, 5, and 8 were evaluated in the new regenerate area using immunohistochemistry. RESULTS: Expressions of BMPs-2,-4 and Smads 1, 5, and 8 were moderate during latency, significantly increased during distraction and decreased towards consolidation period. CONCLUSIONS: The enhanced expression of BMPs and its Smads during distraction compared to the non-distracted group suggests the possible role of BMP signaling pathway in translation of mechanical forces into biological results during DO.

Changes in temporomandibular disc position during growth in young Japanese.

PURPOSE: To detect the initial changes in temporomandibular disc position during growth with MRI examination in a prospective longitudinal study of a young Japanese population. METHODS: Forty-four temporomandibular joints in 22 healthy asymptomatic volunteers, mean age 11.8 years, age range 9-16 years, were examined by MRI. They were re-examined 2-3 years later. The mean age and age range at the second examination were 14.5 years and 12-19 years, respectively. The discs were assessed at three different locations by oblique sagittal planes at the lateral, central and medial parts, and the displacement was evaluated as a partial anterior or total anterior position. RESULTS: Six joints already had a partial anterior position of the disc at the initial examination and no developmental change in disc position was observed during the examination period. Nine joints developed partial disc displacement during the examination period. Every partial displacement was observed at the lateral part of the disc. One joint developed a total anterior position from the lateral to the medial images. CONCLUSIONS: Initial changes in the disc position are likely to start from the lateral part of the joint in the early teenage years and are often asymptomatic.

Intraoperative surgical clearance confirmation of tongue carcinomas using ultrasound.

OBJECTIVES: The existence of tumour cells on the surgical margin can be shown in frozen sections. However, there has been no method to obtain information about surgical clearance intraoperatively. We propose a simple method to confirm the surgical clearance of tongue carcinomas during surgery. METHODS: Just after the resection of a tumour in a conventional manner, the resected fresh specimen was immersed in a gelatine solution, maintaining its original shape and orientation, and refrigerated for solidification of the gelatine. Then ultrasound observation of the gelatine-embedded specimen was performed from the surface of the gelatine. RESULTS: Very fine images could be obtained easily and without any specialist skills. CONCLUSIONS: With this method, the operator can easily obtain information about surgical clearance during the operation.

Configuration of a chiral smectic-C film with a circular inclusion.

It was shown experimentally (P.V. Dolganov et al., Europhys. Lett. 76, 250 (2006)) and by numerical calculations (C. Bohley, R. Stannarius, Eur. Phys. J. E 23, 25 (2007)) that the c -director profile of a two-dimensional chiral smectic-C (SmC) film around a circular inclusion adopts dipolar rather than quadrupolar configuration observed in achiral SmC films. We give an analytical argument on how spontaneous bend inherent in chiral SmC liquid crystals influences the configuration of a SmC liquid crystal film around a circular inclusion imposing tangential anchoring. We find how the angle alpha between two surface defects seen from the center of the inclusion depends on the radius of the inclusion R and the strength of the spontaneous bend q . We show, however, that the contribution of the spontaneous bend to the free energy suffers from mathematical ambiguity; it depends on the mathematical treatment of the outer boundary even when it is at infinity. This might indicate that the shape as well as the treatment of the outer boundary of the film can significantly influence the equilibrium configuration of the c -director and the position of the surface defects.

Functional evaluations of the parotid and submandibular glands using dynamic magnetic resonance sialography.

PURPOSE: To evaluate the functional differences between the parotid and submandibular glands using dynamic MR sialography. METHODS: In 30 volunteers, the time-dependent changes (dynamic changes) in the maximum area of the detectable parotid and submandibular gland ducts on dynamic MR sialographic images were analysed. RESULTS: Dynamic changes in the parotid gland ducts were detectable on MR sialographic images in all volunteers, but images of the submandibular gland ducts were detectable in only 23 volunteers. In addition, the dynamic changes in the submandibular gland ducts in these 23 subjects were less than those seen in the parotid gland ducts. A relationship was found between the changing ratio of parotid (Pearson r=0.448, P=0.013) or submandibular gland ducts (Pearson r=0.418, P=0.047) and the salivary flow rate during the stimulation period. CONCLUSIONS: Dynamic MR sialography allows evaluation of rest and stimulated functioning and morphological evaluation of the parotid and submandibular glands. This technique appears to have many possible applications in the dental, medical and biological fields.

Local delivery system of cytotoxic agents to tumors by focused sonoporation.

Recently, ultrasound-targeting microbubble destruction has been employed in molecular gene therapy, and a new potent nonviral gene transfer method known as 'sonoporation' has been developed. We investigated the efficiency of sonoporation toward growth inhibition of human gingival squamous carcinoma cell line, Ca9-22, in vitro and in vivo. The cytotoxicity of bleomycin (BLM) was investigated using flow-cytometric analysis and Hoechst's staining in vitro assay systems. We found that the delivery of BLM by sonoporation induced cytotoxic effect toward Ca9-22 cells in vitro. Our in vivo results showed that tumors nearly disappeared in Ca9-22 cell-implanted nude KSN/slc mice treated with a low dose of BLM followed by sonoporation during the 4-week experimental period. Histological analysis revealed that the cytotoxic effect was mainly apoptosis. We previously reported that the cytolethal distending toxin B (cdtB) from Actinobacillus actinomycetemcomitans, a periodontopathic bacterium, is responsible for cell cycle arrest and apoptosis in vitro. Thus, we used sonoporation to transfect a cdtB-expressing plasmid into Ca9-22 cells and examined cell viability in vitro and in vivo. We found that an administration of cdtB-expressing plasmid followed by sonoporation-induced marked growth inhibition of Ca9-22 cells and apoptotic cells were also observed in vitro and in vivo. These findings suggest that local administration of cytotoxic agents with sonoporation is a useful method for molecular cancer therapy.

Morphological features of cartilage observed during mandibular distraction in rabbits.

Ossification during distraction osteogenesis can be classified as intramembranous or endochondral. It is not known whether cartilage in the distraction gap is transformed into new bone. The aim of this study was to investigate the morphological features of ossification in the transition of cartilage to bone during mandibular distraction osteogenesis in a rabbit model. A cortical osteotomy was performed and custom-made devices were applied. Immediately after surgery, the devices were lengthened by 0.25 mm every 12h for up 10 days, during which time four rabbits were killed at 0, 5 and 10 days and examined using histological staining and immunohistochemical methods. Apoptotic cells were identified by an in-situ detection assay for nuclear DNA fragmentation using a modified TUNEL procedure, with several sections analyzed using software for histomorphometric analysis. The results showed that the amount of cartilage in the distraction gap was significantly decreased. The cartilage had ossified in two ways, termed endochondral ossification and transchondroid bone formation.

Dynamic magnetic resonance sialography as a new diagnostic technique for patients with Sjögren's syndrome.

OBJECTIVE: To evaluate the clinical utility of dynamic magnetic resonance (MR) sialographic images as a diagnostic tool for patients with Sjögren's syndrome. METHODS: The morphological findings and various kinds of functional parameters in volunteers on dynamic MR sialographic images were compared with those in five patients with definite Sjögren's syndrome. RESULTS: On the MR sialographs of all five patients with Sjögren's syndrome, the so-called 'apple-tree appearance' was seen. The difference in two functional parameters using the dynamic MR sialographic data was elucidated between the two groups. The maximum area of the detectable ducts in the group of patients was significantly smaller (P < 0.001) than that in the group of volunteers. The ratio of change in the detectable ducts in the group of patients was significantly lower (P = 0.011) than that in the group of volunteers. CONCLUSIONS: Our study suggests that dynamic MR sialographic data in addition to MR sialographic images might be useful for the diagnosis of Sjögren's syndrome.

A simple method for bone positioning of mandibular segments.

A simple bone-positioning technique is reported, using a combination of bone plates and a flexible tube filled with acrylic polymer. The flexible tube is solidified by injection of acrylic monomer. This technique is applicable for bone positioning after mandibular resection as well as positioning of the proximal segment after sagittal split ramus osteotomy. This technique has the advantages of greater simplicity and flexibility compared to previously reported methods of bone positioning.

Hepatocyte spheroid formation on a titanium dioxide gel surface and hepatocyte long-term culture.

The cell morphology and expression of differentiated functions of primary rat hepatocytes on a titanium dioxide (TiO(2)) gel surface were investigated. Polystyrene culture dishes were coated with TiO(2) gel by spin-coating an ethanol solution of titanium n-butoxide, hydrolyzing in a humidity chamber and drying with nitrogen gas. The TiO(2) gel layer formed on the polystyrene dishes was transparent and robust, and its surface was quite flat. Rat hepatocytes inoculated on the TiO(2) gel-coated polystyrene dishes gradually accumulated with increasing culture time, and then spontaneously formed many hepatocyte spheroids at 90 +/- 21 microm diameter from about 3 days of culture. The diameter of the spheroids increased during the culture, and was 151 +/- 43 microm at 14 days of culture. Ammonia removal and albumin secretion by hepatocytes on the TiO(2) gel-coated polystyrene dishes were maintained at a high level for at least 14 days of culture compared with on a type I collagen-coated dish and a non-coated polystyrene dish. These results indicate that TiO(2) gel is an adequate material for hepatocyte spheroid formation and long-term culture of spheroids.

Intratumoural expression of thymidylate synthase is an independent predictor of prognosis in patients with squamous cell carcinoma of the tongue: results from a retrospective study.

The aim of this study was to assess the importance of immunohistochemical thymidylate synthase (TS) expression level as a prognostic marker in tongue cancer patients. In 140 patients with primary squamous cell carcinoma (SCC) of the tongue, intratumoural TS expression was evaluated by immunohistochemistry. The level of TS expression was determined by a semiquantitative scoring system, ranging from 1+ to 3+ according to the ratio of TS-positive cells. Of 140 patients, 64 (45.7%), 49 (35.0%) and 27 (19.3%) were assessed as 1+, 2+ and 3+, respectively. Univariate analyses demonstrated that both disease-free survival (DFS) and overall survival (OS) were significantly lower in patients with a TS 3+ tumour than in those with a TS 1+/2+ tumour (DFS: P = 0.0082, OS: P = 0.0100). In a multivariate analysis using the Cox regression model, cervical lymph-node status and TS expression level were selected as independent factors for DFS and OS. Maintenance adjuvant chemotherapy by oral 5-fluorouracil (5-FU) significantly improved DFS and OS in patients with a TS 1+/2+ tumour (DFS: P = 0.0027, OS: P = 0.0398). These data suggest that the level of immunohistochemical TS expression is an independent prognosticator in patients with tongue SCC, and may be useful in the selection of patients who would benefit from oral 5-FU adjuvant chemotherapy.

Stability of the director profile of a nematic liquid crystal around a spherical particle under an external field.

We study numerically the effect of an external magnetic or electric field on the director profiles of a nematic liquid crystal around a spherical particle. We pay particular attention to the stability of a hyperbolic hedgehog defect accompanying the particle, which transforms into a Saturn-ring defect encircling the particle under a sufficiently strong external field. We focus on the particle size dependence of the two important threshold field strengths: the "thermodynamic-transition" field strength H1 at which the hedgehog and the Saturn-ring configurations have the equal free energy, and the critical field strength H2 at which the hedgehog loses its (meta)stability. Our numerical results demonstrate that while H1 is non-monotonically dependent on the particle radius R0, H2 monotonically increases with R0 and the dependence of H2 is weak for large R0. The non-monotonic dependence of H1 on R0 can be explained by comparing the energies of the two configurations and assuming the dependence of those energies on a rescaled field. A crude argument of the energetics of a hyperbolic hedgehog defect under an external field shows that for an asymptotically large R0 the critical field strength is independent of R0, which agrees with our numerical finding.

Velar ascent and morphological factors affecting velopharyngeal function in patients with cleft palate and noncleft controls: a cephalometric study.

This study compares velar ascent and morphological factors affecting velopharyngeal function between patients with repaired cleft palate and noncleft controls from early childhood to puberty. Lateral cephalograms obtained at rest and during blowing from 61 patients with repaired unilateral cleft lip and palate (cleft group) and 82 noncleft controls (control group) were divided into four developmental stages according to age and were studied cross-sectionally. Indices of nasopharyngeal area were derived from a coordinate system and landmarks on lateral cephalograms. The cleft group had lesser velar ascent, more posterosuperior position of the posterior maxilla, shorter velar length, and lesser pharyngeal depth than did the control group. There was a strong correlation between the vertical position of the posterior maxilla and the pharyngeal depth in the cleft group. Discriminant analysis revealed that the cleft group could be discriminated from the control group primarily on the basis of pharyngeal depth, velar length, and velar ascent. Our results suggest that the posterosuperior position of the posterior maxilla in patients with repaired cleft palate, resulting in reduced pharyngeal depth, represents an effort to facilitate velopharyngeal closure by means of shorter velar length and lesser velar ascent.

Utility of three dimension fast asymmetric spin-echo (3D-FASE) sequences in MR sialographic sequences: model and volunteer studies.

OBJECTIVES: To evaluate the utility of 3D-FASE for the visualization of salivary gland ducts for use in MR sialographic sequences. METHODS: We compared MR sialographic images and virtual endoscopic views from 3D-FASE with those from three kinds of sequences described by previous reports in a 3D parotid gland duct model and volunteer. The four sequences were two-dimension fast spin-echo (2D-FSE), three-dimension fast spin-echo (3D-FSE), two-dimension fast asymmetric spin-echo (2D-FASE), and three-dimension fast asymmetric spin-echo (3D-FASE). RESULTS: In the 3D parotid gland duct model, image visibility on visual score was clearest with 3D-FSE, followed by 3D-FASE (P = 0.028). In the volunteers, the visualization of images improved in the following order: 3D-FASE > 3D-FSE > 2D-FSE > 2D-FASE. CONCLUSIONS: The technique of 3D-FASE sequencing is more suitable and useful for MR sialography with an appropriate acquisition time.