Cytological Observations and Bulked-Segregant Analysis Coupled Global Genome Sequencing Reveal Two Genes Associated with Pollen Fertility in Tetraploid Rice

Neo-tetraploid rice with high fertility is a useful germplasm for polyploid rice breeding, which was developed from the crossing of different autotetraploid rice lines. However, little information is available on the molecular mechanism underlying the fertility of neo-tetraploid rice. Here, two contrasting populations of tetraploid rice, including one with high fertility (hereafter referred to as JG) and another with low fertility (hereafter referred to as JD), were generated by crossing Huaduo 3 (H3), a high fertility neo-tetraploid rice that was developed by crossing Jackson-4x with 96025-4x, and Huajingxian74-4x (T452), a low fertility autotetraploid rice parent. Cytological, global genome sequencing-based bulked-segregant (BSA-seq) and CRISPR/Cas9 technology were employed to study the genes associated with pollen fertility in neo-tetraploid rice. The embryo sacs of JG and JD lines were normal; however, pollen fertility was low in JD, which led to scarce fertilization and low seed setting. Cytological observations displayed low pollen fertility (25.1%) and approximately 31.3 and 27.2% chromosome lagging at metaphase I and II, and 28.8 and 24.8% chromosome straggling at anaphase I and II in JD, respectively. BSA-seq of F_2–3 generations and RNA-seq of F₄ generation detected a common fragment, i.e., 18,915,234–19,500,000, at chromosome 7, which was comprised of 78 genes associated with fertility. Among 78 genes, 9 genes had been known to be involved in meiosis and pollen development. Two mutants ny1 (LOC_Os07g32406) and ny2 (LOC_Os07g32040) were generated by CRISPR/Cas9 knockout in neo-tetraploid rice, and which exhibited low pollen fertility and abnormal chromosome behavior. Our study revealed that two unknown genes, LOC_Os07g32406 (NY1) and LOC_Os07g32040 (NY2) play an important role in pollen development of neo-tetraploid rice and provides a new perspective about the genetic mechanisms of fertility in polyploid rice.     

Temburong Bridge,Brunei – Design of two cable stayed bridges

The Temburong Bridge Project is a 30 km long dual two‐lane highway crossing over the Brunei Bay. It will connect the relatively isolated Brunei district of Temburong to the other three Brunei districts. The main objective of the project is to stimulate economic growth in the Temburong region by connecting it to the country's airport and ports. The alignment crosses two navigation channels resulting in the need for two cable stayed bridges – the Brunei Channel Bridge (145 m main span) and the Eastern Channel Bridge (260 m main span). The design of these cable stayed bridges is one of the first applications of the Eurocode to a fully concrete cable stayed bridge. Both cable stayed bridges draw on strong Islamic architectural influences from the region to form a tower shape that is unique and instantly recognisable. Temburong Brücke, Brunei – Entwurf von zwei Schrägkabelbrücken Die Temburong Brücke ist eine 30 km lange Verbindung über die Brunei Bucht mit zwei Richtungsfahrbahnen mit je zwei Fahrstreifen. Sie wird den relativ isolierten Distrikt Temburong mit den anderen drei Distrikten Bruneis verbinden. Hauptanliegen des Projekts ist die Stimulierung von ökonomischem Wachstum in der Temburong Region, indem sie an den Flughafen und Seehafen des Landes angebunden wird. Die Trassierung erfordert die Überquerung zweier Schiffahrtsrouten. Dies wird mit der Anordnung zweier Schrägkabelbrücken ermöglicht, der Brunei Channel Brücke (145 m Hauptspannweite) und der Eastern Channel Brücke (260 m Hauptspannweite). Die Bemessung der beiden Brücken stellt eine der ersten Anwendungen des Eurocode auf Schrägkabelbrücken mit Betonüberbau dar. Optisch sind die beiden Brücken geprägt von der islamischen Architektur der Umgebung, die Pylonform ist ein besonderes Erkennungsmerkmal der Brücken.     

Epstein-Barr virus-positive multiple early gastric cancers and dysplastic lesions: a case report

Epstein-Barr virus (EBV) has been implicated as a causal virus of gastric cancer with episomal monoclonality, elevated antibodies and a unique morphologic expression in the early intramucosal stage, but the infection mechanisms have not been demonstrated. EBV has been shown only in the cancerous lesions by the highly sensitive EBV-encoded small RNA in situ hybridization (EBER-ISH) method, not in the dysplastic mucosa adjacent to the cancer. A case is presented of multiple EBV-positive gastric cancer and dysplastic epithelium observed in a 52-year-old man. Serial cut sections of the gastrectomy specimen showed four small cancerous lesions, three of which were EBER-positive, and three EBER-positive, minute, non-cancerous dysplastic lesions. The three cancerous lesions were intramucosal cancer, with one having minimal submucosal invasion forming a lymphoepithelioma-like histology. All of these EBER-positive cancerous and dysplastic lesions showed intense CD8 T-lymphocytic infiltration. There was no such findings in the EBV-negative cancerous lesion. It was concluded that EBV infection may occur in the epithelial cells of atrophic gastric mucosa, and progress to cancer with monoclonal expansion through the EBV-positive dysplastic change. Cytotoxic T-lymphocytic reactions can occur even in the dysplastic lesions. Multifocal EBV infection in the gastric mucosa may occur and, if necessary, total gastrectomy is recommended in such a case.     

Modern interpretation on the high-stretching of natural rubber attained by the classic ‘racking’ method

To investigate the mechanism of high elongation of natural rubber attained by the ‘racking method’, a strip of smoked-sheet was elongated up to 150 times in length, by stretching rapidly at room temperature and forcing to shrink at 75-80 °C repeatedly. On X-ray diffraction, a typical fibre pattern with an amorphous ring appeared already at ×10 (stretch ratio=10). The degree of crystallinity increased to the level of 17-18% at ×20 and turned to decrease after ×60, but the degree of crystallite orientation reached at a high level already at ×10 and did not change significantly and the half-height width of reflection profiles stayed almost at a constant level, while the Young's modulus increased up to ca. 300 MPa at ×100 and then fell rather discontinuously. It was assumed that some fractions of chain segments between entanglements were gradually broken in each step of stretching operation, rather than disentangled and slipped.     

Metabolomic Profiling of Plasma from Melioidosis Patients Using UHPLC-QTOF MS Reveals Novel Biomarkers for Diagnosis

To identify potential biomarkers for improving diagnosis of melioidosis, we compared plasma metabolome profiles of melioidosis patients compared to patients with other bacteremia and controls without active infection, using ultra-high-performance liquid chromatography-electrospray ionization-quadruple time-of-flight mass spectrometry. Principal component analysis (PCA) showed that the metabolomic profiles of melioidosis patients are distinguishable from bacteremia patients and controls. Using multivariate and univariate analysis, 12 significant metabolites from four lipid classes, acylcarnitine (n = 6), lysophosphatidylethanolamine (LysoPE) (n = 3), sphingomyelins (SM) (n = 2) and phosphatidylcholine (PC) (n = 1), with significantly higher levels in melioidosis patients than bacteremia patients and controls, were identified. Ten of the 12 metabolites showed area-under-receiver operating characteristic curve (AUC) >0.80 when compared both between melioidosis and bacteremia patients, and between melioidosis patients and controls. SM(d18:2/16:0) possessed the largest AUC when compared, both between melioidosis and bacteremia patients (AUC 0.998, sensitivity 100% and specificity 91.7%), and between melioidosis patients and controls (AUC 1.000, sensitivity 96.7% and specificity 100%). Our results indicate that metabolome profiling might serve as a promising approach for diagnosis of melioidosis using patient plasma, with SM(d18:2/16:0) representing a potential biomarker. Since the 12 metabolites were related to various pathways for energy and lipid metabolism, further studies may reveal their possible role in the pathogenesis and host response in melioidosis.     

Real-time investigation of crystallization in poly(vinylidene fluoride)-based nano-composites probed by infrared spectroscopy

Via time-resolved Fourier transform infrared spectroscopy (FTIR), we examined the real-time investigation of the conformational changes of poly(vinylidene fluoride) (PVDF) chain segment during crystallization of neat PVDF and the corresponding nano-composites having intercalated structure. It was shown that in the following crystallization processes the crystal growth was virtually the same in both nano-composites and neat PVDF. We have examined an annealing at an infinitely long time at 200 °C (∼20 min) to erase the thermal history in the nano-composites. The dispersed titanate nano-filler particles exhibited strong contribution to enhance the heterogeneous nucleation for the formation of both γ- and β-phase crystals.