Change and Impact of MicroRNA Modification With Age in Drosophila Melanogaster

microRNAs (miRNAs) are 20~24nt small RNAs that are critical for many biological aspects, from development to age-associated processes. Starting from the identification of the first miRNA, lin-4, hundreds of miRNAs have been discovered across species. To reveal the role of miRNAs in aging, studies have profiled changes in miRNA levels with age. However, increasing evidence suggests that miRNAs show heterogeneity in length and sequence in different biological contexts. Despite the observation of such heterogeneity, it is largely unknown how such heterogeneity is generated, and whether it is biologically regulated or important. Here we report the characterization of a novel 3\u27-to-5\u27 exonuclease, Nibbler (Nbr), that generates different length miRNA isoforms in Drosophila. Small RNA deep-sequencing in flies followed by northern blots revealed a set of miRNAs that depend on Nbr for their length patterns. We pursued the biological importance of nbr by analyzing the impact of loss-of-function mutation of the nbr gene, as well as the effect of nbr loss on other classes of small RNAs such as piRNAs and endo-siRNAs by small RNA deep-sequencing of ovary tissue. This analysis revealed a critical role of nbr to regulate age-associated traits such as brain degeneration and locomotion, as well as the length of piRNAs and endo-siRNAs. Detailed analysis revealed that, even though most piRNA loci are affected, only select endo-siRNA loci are affected upon nbr loss. Finally, we report a novel age-associated change in miRNA heterogeneity: the increase in 2\u27-O-methylation of select miRNA isoforms with age. Detailed study by immunoprecipitation of Ago1 and Ago2, followed by northern blots and RNA deep-sequencing showed that this reflects increased loading of miRNA isoforms into Ago2 (siRISC) with age. Importantly, the loss of 2\u27-O-methylation of small RNAs by hen1 and ago2 mutations led to accelerated brain degeneration and shorter lifespan. Together, this research has revealed an important mechanism to generate 3\u27 end heterogeneity of miRNAs by Nbr, and a novel age-associated heterogeneity of miRNAs, 2\u27-O-methylation of miRNAs. Importantly, the lack of such heterogeneity led to accelerated age-associated defects, implying the potential importance of regulating the 3\u27 end heterogeneity of small RNAs on age-dependent processes in Drosophila

Results of the Archaeological Project at Ak Beshim (Suyab), Kyrgyz Republic from 2011 to 2013 and a Note on the Site`s Abandonment

The National Research Institute for Cultural Properties, Tokyo has been conducting an archaeological project at Ak Beshim (Suyab) in the Chuy Valley, Kyrgyz Republic since 2011. This paper provides preliminary results of the project`s first three seasons from 2011 to 2013. The excavations revealed a Qarakhanid main street and several houses along the street in the center of Shakhrsitan of Ak Beshim. This layer is dated by radiocarbon to the late 10th century. According to historical documents, the region was Islamized in the middle of the 10th century, so this layer provides relevant cultural materials for studying Islamization in this region. Radiocarbon dating also suggests that Ak Beshim was abandoned in the late 10th century, 200 years earlier than previously believed. Reasons behind the abandonment of Ak Beshim are considered at the end of this paper

OBSERVATION AND ANALYSIS OF THE A1-A2 SPLITTING OF CH3D

Sub-Doppler resolution spectroscopy of CH$_{3}$D has been carried out for the $nu _1$ and $nu _4$ fundamental bands using a comb-referenced difference-frequency generation spectrometer. Thirty transitions from the low-$J''$ and $K'' = 3$ levels are observed with a resolution of 60 to 100 kHz, and the $A_1$-$A_2$ splitting is resolved for twenty-three of the thirty transitions. Most of them are overlapped in Doppler broadening and resolved for the first time, as far as we know. The absolute transition frequencies are determined with a typical uncertainty of 4 kHz. The $A_1$-$A_2$ splitting constant of the $K'' = 3$ levels is yielded as $2h_{3,v=0} = (1.5641 pm 0.0026)$ Hz for the ground vibrational state. Those of the $K' = 3$ levels for the $v_1 = 1$ states and of the ($K' = 2$, $l = -1$) and ($K' = 4$, $l = 1$) levels for the $v_4 = 1$ state are also determined including the $J'$-dependent terms

The Development of Urbanism and Pastoral Nomads in the Southern Levant -Chalcolithic and Early Bronze Age Stone Tool Production Industries and Flint Mines in the Jafr Basin, Southern Jordan-

'The development of urbanism' has been one of the most important topics since V. G Childe's seminal works. This paper will discuss the impact of the development of urbanism III the Southern Levant on pastoral nomads using archaeological data from the Jafr Basin, Southern Jordan. The Jafr Basin is one of the best flint sources in the Southern Levant and yields high quality Eocene flint. In the Early Bronze Age when a number of fortified urban settlements appeared in the Southern Levant, pastoral nomads in the Jafr Basin started intensive flint mining and stone tool production of tabular scrapers. Tabular scrapers were distributed from the basin in large quantities to urban settlements and farming villages. Pastoral nomads in the Negev and Sinai also started utilizing desert resources for urban settelements. It is likely that in the Early Bronze Age the arid areas were economically integrated with the moister zones to a greater degree than before. The development of urbanism had the effect of making pastoral nomads more to\\l1 and market-oriented. In the Early Bronze Age, a variety of desert products were distributed to sedentary settlements by pastoral nomads. Meanwhile, Early Bronze Age pastoral nomads probably became more dependent for living necessities and luxuries such as cereals, foods, vegetables and clothes on markets in urban communities

Relationship between radiographic grading of osteoarthritis and the biochemical markers for arthritis in knee osteoarthritis

The aim of this study was to investigate the relationship between the biochemical markers of arthritis and the radiographic grading of osteoarthritis (OA) in knees. Seventy-one women aged 49–85 years with knee OA were studied. Anterior–posterior knee radiographs and hand radiographs were taken in all patients. The radiographic grading of OA in the knee was performed by using the Kellgren–Lawrence criteria and the joint space width. The 71 patients with knee OA were divided into two groups: 37 patients exhibiting generalized osteoarthritis (GOA) and 34 non-GOA patients, according to the grading of their hand radiograph. C-reactive protein (CRP), urinary pyridinoline, YKL-40, plasma matrix metalloproteinase (MMP)-3, MMP-9 and tissue inhibitor of metalloproteinases (TIMP)-1 were measured as the biochemical markers of arthritis. The radiographic grading with the Kellgren–Lawrence scale revealed a significant relationship to the joint space width (P = 0.003): the joint space width decreased with increasing Kellgren–Lawrence grade. All biochemical markers had negative correlations with the joint space width, but only urinary pyridinoline had a significant correlation (P = 0.039). Pyridinoline (P = 0.034) and TIMP-1 (P = 0.017) also exhibited a significant relationship to the Kellgren–Lawrence grade. In GOA evaluations, the joint space width did not differ between GOA and non-GOA patients. CRP, pyridinoline, YKL-40 and MMP-3 levels were significantly greater in GOA patients than in non-GOA patients. CRP, pyridinoline, YKL-40, MMP-3 and TIMP-1 levels each related to at least one of the radiographic gradings. Furthermore, pyridinoline related to every type of radiographic grading examined in the present study

Immunohistochemical Analysis of Oral Dysplasia: Diagnostic Assessment by Fascin and Podoplanin Expression

The aim of this study was to investigate fascin and podoplanin expression in oral dysplasia and carcinoma in situ (CIS) immunohistochemically, and to evaluate their relationship to histopathological diagnosis based on architectural and cytological features. Fascin and podoplanin expression patterns were analyzed immunohistologically in 26 specimens of oral lesions, including benign disease (hyperplasia, papilloma, and others), intraepithelial neoplasia/borderline disease (dysplasia), and malignant disease (CIS, invasive squamous cell carcinoma). Fascin expression was scored into four original categories, and podoplanin expression was scored into five previously established categories. The relationship between the immunohistochemically determined scores of fascin and podoplanin expression and the architectural and cytological features in the hematoxylin-eosin-stained slides was analyzed statistically. The immunostaining scores for fascin and podoplanin were significantly higher in dysplasia and CIS than in benign disease (p=0.0011, p=0.00036), and they were significantly higher in dysplasia than in benign disease (p=0.0087, p=0.0032). In all cases of invasive SCC, fascin was expressed mainly in the cytoplasm of the tumor cells and fascin expression extended from the destruction of the basal layer of the epithelium to the upper layer of the epithelium and podoplanin was expressed in the cytoplasm and membrane of the tumor cells. This was the first report of up-regulation of fascin in oral dysplasia. Our results suggest that it would be helpful for improving the diagnostic accuracy of oral dysplasia and CIS to assess the expression of fascin and podoplanin immunohistochemically