Use of a YAP1 overexpression cassette conferring specific resistance to cerulenin and cycloheximide as an efficient selectable marker in the yeast Saccharomyces cerevisiae.

Drug-resistance markers for yeast transformation are useful because they can be applied to strains without auxotrophic mutations. However, they are susceptible to technical difficulties, namely lower transformation efficiency and the appearance of drug-resistant mutants without the marker. To avoid these problems, we have constructed a phosphoglycerate kinase (PGK) promoter-driven YAP1 expression cassette, called PGKp-YAP1. Yeast cells containing PGKp-YAP1 were resistant to cycloheximide, a protein synthesis inhibitor, and also to cerulenin, a fatty acid synthesis inhibitor, but not to other drugs tested. The transformation efficiency of PGKp-YAP1 using cerulenin selection was comparable to that using a URA3 auxotrophic marker when low concentrations of cerulenin were used. Non-transformed drug-resistant colonies did appear on the low-concentration cerulenin plates. However, these non-transformed colonies could easily be identified, based on their cycloheximide sensitivity and/or their resistance to aureobasidin A to which the transformants were sensitive. Therefore, the dual drug resistance of PGKp-YAP1 could be used as an effective selection for PGKp-YAP1 recipient cells. The PGKp-YAP1 marker was used to disrupt the LYS2 gene and to transform an industrial yeast strain, indicating that this marker can be used for efficient and reliable gene manipulations in any Saccharomyces cerevisiae strain.     

Hybrid Link/Path-Based Design for Translucent Photonic Network Dimensioning

The advancement of ultralong-haul transmission technology has dramatically enhanced the all-optical reaches. However, the actual situations of installed fiber and sites for terrestrial network often prevent implementing a purely transparent network, and thus, opaque reshaping retiming regenerating (3R) regeneration is required to guarantee optical signal reachability. Since 3R regenerators based on optical/electrical/optical conversion tend to dominate the total network costs, an efficient network design method that allocates a minimum number of 3R regenerators to optimum locations is essential to build a cost-effective photonic network. In this paper, we propose such a network-dimensioning method by combining the advantages of link-based and path-based design approaches. It first guarantees optical signal reachability for any possible traffic demand in each segmented linear link. After combining all the links, excessive regenerators are eliminated based on the optical signal quality check with -factor calculation for each wavelength path. A trial design of a large-scale mesh network demonstrated a significant cost savings of more than 30% in comparison with a conventional link-based design. In the trial, the impact of fiber loss coefficient over the total network cost was investigated quantitatively, addressing the importance of such quantitative modeling and analysis.     

Development of an epileptic seizure prediction algorithm using R–R intervals with self-attentive autoencoder

Epilepsy is a neurological disorder that may affect the autonomic nervous system (ANS) from 15 to 20 min before seizure onset, and disturbances of ANS affect R–R intervals (RRI) on an electrocardiogram (ECG). This study aims to develop a machine learning algorithm for predicting focal epileptic seizures by monitoring R–R interval (RRI) data in real time. The developed algorithm adopts a self-attentive autoencoder (SA-AE), which is a neural network for time-series data. The results of applying the developed seizure prediction algorithm to clinical data demonstrated that it functioned well in most patients; however, false positives (FPs) occurred in specific participants. In a future work, we will investigate the causes of FPs and optimize the developing seizure prediction algorithm to further improve performance using newly added clinical data.

     

Comparison of span configurations of Raman-amplifieddispersion-managed fibers

We evaluate the performance of dispersion-managed fiber transmission systems employing distributed Raman amplification with 50- and 100-km span length for three different span configurations. The evaluation considers the optical signal-to-noise ratio and the nonlinear phase shift as a measure for the impact of nonlinear effects. The simulation results indicate that a 100-km-long span with the dispersion compensating fiber placed in the span center achieves the best tradeoff between optical signal-to-noise degradation and nonlinear effects     

BER estimation in optical fiber transmission systems employing all-optical 2R regenerators

A bit error rate estimation method for optical fiber transmission systems employing all-optical 2R regenerators is presented. The method is based on the linearization of the nonlinear transfer function of the regenerators in three sections. Using this linearized three-section model, the impact of 2R regeneration on the system performance as a function of the extinction ratio, the clamping factor, and the steepness of the transfer function is studied.     

Detection of a point mutation in FAS2 gene of sake yeast strains by allele-specific PCR amplification

To identify yeast mutants with a point mutation, detection of the specific mutant alleles is necessary. For this purpose, we applied allele-specific polymerase chain reaction (PCR) to detect the FAS2-1250S dominant mutant allele that encodes an altered fatty acid synthase in Japanese brewer's yeast strains. These strains are known to produce a higher amount of ethyl caproate in Japanese sake. The mutant strains were supposed to be diploid and to contain heterozygous alleles, including wild-type FAS2 and a dominant FAS2-1250S. A set of oligonucleotide primers was designed to contain different nucleotides at their 3' termini: one type was identical to the wild type and the other to the mutant FAS2. Another set of primers was designed to have an additional mismatch at the second nucleotide from their 3' termini. By testing with control strains, we established PCR conditions for specific amplification. Using these conditions and a simple template preparation procedure with SDS, the presence of the allele was detected in commercially used sake yeast strains. The method presented here will be useful for the identification of specific yeast strains.     

Neoadjuvant intraarterial chemotherapy for ten cases of inflammatory breast cancer by Seldinger's methods

The conventional methods (CM) of intraarterial (IA) chemotherapy for inflammatory breast cancer were catheterizations from superior epigastric and brachial artery under general anesthesia. Since 1997, we selected the Seldinger's methods (SM) for ten cases of the disease to control local effects and simplify the technique. Complications of the SM were slight, and side effects were equal to those with CM. The postoperative pathological findings of the SM showed the direct effects of chemotherapy to tumor cells (degenerative and necrotic changes) as compared with the embolism-like effects of CM. But the overall histological effects of chemotherapy by SM were almost equal to those for CM. The strong points of the SM were as follows: 1) More selective IA chemotherapy is available, 2) one can find the passage of the aim vessels and no trouble related to catheter, 3) general anesthesia is not necessary, and the techniques are simple, 4) the wounds are not remarkable. The disadvantages are as follows: 1) Patients must rest the day of IA chemotherapy, 2) in 20% of the procedures, one can not search the vessels, and 3) in 4 cases complications of stiffness of Mj or Mn pectral muscles were found. In future, we expect more effective results by dose escalation or combination chemotherapy.     

Inflammatory pseudotumor of the abdomen: report of a case

BACKGROUND: Cosmetic denervation of hyperfunctional facial lines using botulinum toxin (Botox, Allergan, Inc., Irvine, CA) has gained growing popularity over recent years. Understanding the clinical use and effects of botulinum toxin requires a thorough understanding of the muscular anatomy of the treatment areas. OBJECTIVE: The purpose of this article is to review the anatomy of the frown, forehead, and periocular regions. Function of individual muscles is discussed to understand proper injection technique. CONCLUSIONS: The anatomy of the frown, forehead, and periocular regions is complex. Individual muscles are tightly intertwined and treatment of one anatomic region may affect many different muscles. A complete understanding of the anatomy of the upper face is essential to ensure proper injection technique, safe and predictable results as well as anticipating complications.     

YHp as a highly stable,hyper-copy,hyper-expression plasmid constructed using a full 2-μm circle sequence in cir0 strains of Saccharomyces cerevisiae

In the yeast Saccharomyces cerevisiae, the yeast episomal plasmid (YEp), containing a partial sequence from a natural 2-μm plasmid, has been frequently used to induce high levels of gene expression. In this study, we used Japanese sake yeast natural cir⁰ strain as a host for constructing an entire 2-μm plasmid with an expression construct using the three-fragment gap-repair method without Escherichia coli manipulation. The 2-μm plasmid contains two long inverted repeats, which is problematic for the amplification by polymerase chain reaction. Therefore, we amplified it by dividing into two fragments, each containing a single repeat together with an overlapping sequence for homologous recombination. TDH3 promoter-driven yEmRFP (TDH3p-yEmRFP) and the URA3 were used as a reporter gene and a selection marker, respectively, and inserted at the 3′ end of the RAF1 gene on the 2-μm plasmid. The three fragments were combined and used for the transformation of sake yeast cir⁰ ura3^- strain. The resulting transformant colonies showed a red or purple coloration, which was significantly stronger than that of the cells transformed with YEp-TDH3p-yEmRFP. The 2-μm transformants were cultured in YPD medium and observed by fluorescence microscopy. Almost all cells showed strong fluorescence, suggesting that the plasmid was preserved during nonselective culture conditions. The constructed plasmid maintained a high copy state similar to that of the natural 2-μm plasmid, and the red fluorescent protein expression was 54 fold compared with the chromosomal integrant. This vector is named YHp, the Yeast Hyper expression plasmid.     

Subharmonic hybrid mode-locking of a monolithic semiconductor laser

Performance of a subharmonically hybrid mode-locked (SH-ML) monolithic semiconductor laser is investigated. A 33-GHz passively mode-locked distributed Bragg reflector semiconductor laser is stabilized by the injection of an electrical signal with a subharmonic frequency of the laser cavity resonance. Systematic measurements on the phase noise, timing jitter, amplitude modulation, and locking bandwidth are performed for the second- and third-order SH-ML conditions, and the results are compared with the fundamental hybrid mode-locking (FH-ML) case. Low timing jitter of less than 0.6 ps, comparable to that under the FH-ML case, is achieved for the both SH-ML cases. The amplitude modulation imposed by the subharmonic driving frequencies is found to be very small (<-24 dBc) for the second-order SH-ML because of the sufficiently low modulation response of the laser at 16.5 GHz. The third-order SH-ML is found to exhibit a very unique locking characteristics, leading to a maximum locking bandwidth of 56 MHz that is even larger than that for the FH-ML case