Effect of pharmacological inhibition of the pontine respiratory group on swallowing interneurons in the dorsal medulla oblongata.

OBJECTIVES: To examine the role of neurons of the pontine respiratory group (PRG) overlapping with the Kölliker-Fuse nucleus in the regulation of swallowing, we compared the activity of swallowing motor activities and interneuron discharge in the dorsal swallowing group in the medulla before and after pharmacological inhibition of the PRG. METHODS: In 23 in situ perfused brainstem preparation of rats, we recorded the activities of the vagus (VNA), hypoglossal (HNA), and phrenic nerves (PNA), and swallowing interneurons of the dorsal medulla during fictive swallowing elicited by electrical stimulation of the superior laryngeal nerve or oral water injection. Subsequently, respiratory- and swallow-related motor activities and single unit cell discharge were assessed before and after local microinjection of the GABA-receptor agonist muscimol into the area of PRG ipsilateral to the recording sites of swallowing interneurons. RESULTS: After muscimol injection, the amplitude and duration of swallow-related VNA bursts decreased to 71.3 ± 2.84 and 68.1 ± 2.76 % during electrically induced swallowing and VNA interburst intervals during repetitive swallowing decreased. Similar effects were observed for swallowing-related HNA. The swallowing motor activity was similarly qualitatively altered during physiologically induced swallowing. All 23 neurons were changed in either discharge duration or frequency after PRG inhibition, however, the general discharge patterns in relation to the motor output remained unchanged. CONCLUSION: Descending synaptic inputs from PRG provide control of the primary laryngeal sensory gate and synaptic activity of the PRG partially determine medullary cell and cranial motor nerve activities that govern the pharyngeal stage of swallowing.

Development of Artificial System to Induce Chromatin Loosening in Saccharomyces cerevisiae.

In eukaryotic cells, loosening of chromatin causes changes in transcription and DNA replication. The artificial conversion of tightly packed chromatin (heterochromatin) to loosely packed chromatin (euchromatin) enables gene expression and regulates cell differentiation. Although some chemicals convert chromatin structures through histone modifications, they lack sequence specificity. This study attempted to establish a novel technology for inducing chromatin loosening in target regions of Saccharomyces cerevisiae. We focused on histone acetylation, which is one of the mechanisms of euchromatin induction. The sequence-recognizing ability of the dead Cas9 (dCas9) and guide RNA (gRNA) complex was used to promote histone acetylation at a targeted genomic locus. We constructed a plasmid to produce a fusion protein consisting of dCas9 and histone acetyltransferase Gcn5 and a plasmid to express gRNA recognizing the upstream region of heterochromatic URA3. Confocal microscopy revealed that the fusion proteins were localized in the nucleus. The yeast strain producing the fusion protein and gRNA grew well in the uracil-deficient medium, while the strain harboring empty plasmids or the strain containing the mutations that cause loss of nucleosomal histone acetylation activity of Gcn5 did not. This suggests that the heterochromatin was loosened as much as euchromatin through nucleosomal histone acetylation. The amount of euchromatic DNA at the target locus increased, indicating that chromatin loosening was induced by our system. Nucleosomal histone acetylation in heterochromatic loci by our developed system is a promising method for inducing euchromatic state in a target locus.

Firing characteristics of swallowing interneurons in the dorsal medulla during physiologically induced swallowing in perfused brainstem preparation in rats.

Oropharyngeal swallowing is centrally mediated by a swallowing central pattern generator (Sw-CPG) in the medulla oblongata. The activity of the Sw-CPG depends on the sensory inputs determined by physical and chemical bolus properties. Here we investigate the sensory-motor integration during swallowing arising from different sensory sources. To do so we electrically stimulated the superior laryngeal nerve and we triggered swallowing with oral injections of distilled water or capsaicin solution and extracellularly recorded from swallowing interneurons in arterially perfused brainstem preparations of rats. We recorded the activities of 40 neurons, while monitoring the motor activities of the phrenic, vagal and hypoglossal nerves. Eighteen neurons responded to electrical stimulation of the ipsilateral superior laryngeal nerve, and 6 neurons were excited by oral fluid injection, while 16 non-respiratory neurons did not receive afferent inputs to either electrical or physiological stimuli. The cellular activities displayed by swallowing interneurons during electrical and physiological stimulation of pharyngeal and laryngeal afferent input reveal complex adaptations of the timing of firing patterns and frequencies. The modulation of neuronal activity is likely to contribute to the coordination of efficient bolus transfer during the pharyngeal stage of swallowing.

Synthesis of substituted anilines via a gold-catalyzed three-component reaction.

A three-component reaction for the synthesis of substituted anilines by a gold(i)-catalyzed domino reaction was developed. Cationic gold catalysts selectively and sequentially activated two different alkynes, which were involved in pyrrole synthesis and subsequent Diels-Alder reaction. The sequential formal (3 + 2) annulation/Diels-Alder reaction of three components provided a variety of substituted anilines in a modular fashion. Moreover, utility of the aniline products was demonstrated by derivatization to substituted benzoxazines, which are pharmaceutically important heterocycles.

The long-term patency of a gastroepiploic artery bypass graft deployed in a semiskeletonized fashion: predictors of patency.

OBJECTIVES: Whether or not using the gastroepiploic artery (GEA) is associated with improved outcomes of coronary artery bypass grafting (CABG) remains unclear. Previous research has shown that the short-term function of the GEA was strongly associated with the degree of native vessel stenosis. We assessed the association between long-term GEA patency and the degree of stenosis of the coronary artery. METHODS: We retrospectively examined 517 patients who underwent CABG with an in situ semiskeletonized GEA from January 2000 to January 2015. In this cohort, 282 (54.5%) patients underwent distant radiological evaluations for >1 year post-surgery (range 1-18 years after surgery). Quantitative coronary angiography was used to measure the degree of stenosis of the native coronary artery. Preoperative angiographic parameters include the minimal lumen diameter (MLD) and the percentage of target vessel stenosis. A multivariable stepwise Cox proportional hazards regression analysis was used to identify predictors of angiographic occlusion. RESULTS: The cumulative patency rate of the GEA was 79.3% at 10 years. A multivariable analysis showed that an MLD (hazard ratio 4.43, 95% confidence interval 3.25-6.82; P < 0.001) was an independent risk factor of GEA occlusion. A time-dependent receiver operating characteristic (ROC) curve analysis identified that an MLD >1 mm was set as the cut-off value for graft occlusion. Patients with an MLD <1 mm had a 10-year patency rate of 89.8%. CONCLUSIONS: The long-term patency of the semiskeletonized GEA was acceptable. The target vessel MLD obtained using quantitative coronary angiography was a strong predictor of patency. Good long-term patency can be expected for an MLD <1 mm.

On-chip ultrasonic manipulation of microparticles by using the flexural vibration of a glass substrate.

As biotechnology develops, techniques for manipulating and separating small particles such as cells and DNA are required in the life sciences. This paper investigates on-chip manipulation of microparticles in small channels by using ultrasonic vibration. The chip consists of a rectangular glass substrate with a cross-shaped channel (cross-section: 2.0×2.0mm2) and four lead zirconate titanate transducers attached to the substrate's four corners. To efficiently generate the flexural vibration mode on the chip, we used finite element analysis to optimize the configurations of the glass substrate and transducers. Silicon carbide microparticles with an average diameter of 50µm were immersed in the channels, which were filled with ethanol. By applying an in-phase input voltage of 75V at 225kHz to the four transducers, a flexural vibration mode with a wavelength of 13mm was excited on the glass substrate, and this flexural vibration generated an acoustic standing wave in the channel. The particles could be trapped at the nodal lines of the standing wave. By controlling the driving phase difference between the two pairs of transducers, the vibrational distribution of the substrate could be moved along the channels so that the acoustic standing wave moved in the same direction. The trapped particles could be manipulated by the two-phase drive, and the transport direction could be switched at the junction of the channels orthogonally by changing the combination of the driving condition to four transducers.

Dispersions of image potential states on surfaces of clean graphite and lead phthalocyanine film.

Dispersions of image potential states on a graphite surface (denoted IPS1) and on 1 monolayer (ML) film (denoted IPS2) of lead phthalocyanine (PbPc) are investigated by the micro-spot angle-resolved two-photon photoemission (micro-AR-2PPE) spectroscopy. On the graphite surface, whole dispersions of the two members of IPS1 (n = 1 and 2) are observed. The n = 1 IPS1 peak is weakly visible at energy higher than the vacuum level. The effective mass of an electron in the n = 1 IPS1 becomes slightly light at the high momentum region, suggesting the interaction between the IPS1 and the unoccupied σ-band of graphite. On the PbPc film, the IPS2 band forms a band gap and back-folds at the boundary of the Brillouin zone. A 1-dimensional Kronig-Penny model is used to reproduce the effective mass and the shift of binding energy.

A Case Report of a Malignant Fibrous Histiocytoma in a T-cell Receptor ß Chain and p53 Double-knockout Mouse.

A subcutaneous tumor was found in the right abdomen of a 16-week-old male TCRß and p53 double-knockout mouse. The tumor had indistinct borders with the surrounding tissue. The cut surface after formalin fixation was pale yellowish white, partially dark red and partly white. Histologically, the tumor was composed of three distinct regions. The first region showed pleomorphic cells arranged in sheets. The second region showed spindle cells arranged in interlacing fascicles. The final region contained a mixture of the above mentioned two types of cells. Furthermore, a small amount of collagen fibers, round cells, multinucleated giant cells, and cells with eosinophilic granules were observed between these tumor cells. Immunohistochemical examination and electron microscopy identified that the pleomorphic cells and spindle cells were histiocytes and fibroblasts, respectively, and that the round cells were undifferentiated mesenchymal cells. Based on these findings, the tumor was diagnosed as a malignant fibrous histiocytoma.