The in vitro real-time oscillation monitoring system identifies potential entrainment factors for circadian clocks

BACKGROUND: Circadian rhythms are endogenous, self-sustained oscillations with approximately 24-hr rhythmicity that are manifested in various physiological and metabolic processes. The circadian organization of these processes in mammals is governed by the master oscillator within the suprachiasmatic nuclei (SCN) of the hypothalamus. Recent findings revealed that circadian oscillators exist in most organs, tissues, and even in immortalized cells, and that the oscillators in peripheral tissues are likely to be coordinated by SCN, the master oscillator. Some candidates for endogenous entrainment factors have sporadically been reported, however, their details remain mainly obscure. RESULTS: We developed the in vitro real-time oscillation monitoring system (IV-ROMS) by measuring the activity of luciferase coupled to the oscillatory gene promoter using photomultiplier tubes and applied this system to screen and identify factors able to influence circadian rhythmicity. Using this IV-ROMS as the primary screening of entrainment factors for circadian clocks, we identified 12 candidates as the potential entrainment factor in a total of 299 peptides and bioactive lipids. Among them, four candidates (endothelin-1, all-trans retinoic acid, 9-cis retinoic acid, and 13-cis retinoic acid) have already been reported as the entrainment factors in vivo and in vitro. We demonstrated that one of the novel candidates, 15-deoxy-Δ(12,14)-prostaglandin J(2 )(15d-PGJ(2)), a natural ligand of the peroxisome proliferator-activated receptor-γ (PPAR-γ), triggers the rhythmic expression of endogenous clock genes in NIH3T3 cells. Furthermore, we showed that 15d-PGJ(2 )transiently induces Cry1, Cry2, and Rorα mRNA expressions and that 15d-PGJ(2)-induced entrainment signaling pathway is PPAR-γ – and MAPKs (ERK, JNK, p38MAPK)-independent. CONCLUSION: Here, we identified 15d-PGJ(2 )as an entrainment factor in vitro. Using our developed IV-ROMS to screen 299 compounds, we found eight novel and four known molecules to be potential entrainment factors for circadian clocks, indicating that this assay system is a powerful and useful tool in initial screenings

The role of ARX in human pancreatic endocrine specification

The in vitro differentiation of human embryonic stem cells (hESCs) offers a model system to explore human development. Humans with mutations in the transcription factor Aristaless Related Homeobox (ARX) often suffer from the syndrome X-linked lissencephaly with ambiguous genitalia (XLAG), affecting many cell types including those of the pancreas. Indeed, XLAG pancreatic islets lack glucagon and pancreatic polypeptide-positive cells but retain somatostatin, insulin, and ghrelin-positive cells. To further examine the role of ARX in human pancreatic endocrine development, we utilized genomic editing in hESCs to generate deletions in ARX. ARX knockout hESCs retained pancreatic differentiation capacity and ARX knockout endocrine cells were biased toward somatostatin-positive cells (94% of endocrine cells) with reduced pancreatic polypeptide (rarely detected), glucagon (90% reduced) and insulin-positive (65% reduced) lineages. ARX knockout somatostatin-positive cells shared expression patterns with human fetal and adult δ-cells. Differentiated ARX knockout cells upregulated PAX4, NKX2.2, ISL1, HHEX, PCSK1, PCSK2 expression while downregulating PAX6 and IRX2. Re-expression of ARX in ARX knockout pancreatic progenitors reduced HHEX and increased PAX6 and insulin expression following differentiation. Taken together these data suggest that ARX plays a key role in pancreatic endocrine fate specification of pancreatic polypeptide, somatostatin, glucagon and insulin positive cells from hESCs

Ablation of the scaffold protein JLP causes reduced fertility in male mice

金沢大学がん研究所がん分子細胞制御The specific and efficient activation of mitogen-activated protein kinase (MAPK) signaling modules is mediated, at least in part, by scaffold proteins. c-Jun NH2-terminal kinase (JNK)-associated leucine zipper protein (JLP) was identified as a scaffold protein for JNK and p38 MAPK signaling modules. JLP is expressed nearly ubiquitously and is involved in intracellular signaling pathways, such as the Gα13 and Cdo-mediated pathway, in vitro. To date, however, JLP expression has not been analyzed in detail, nor are its physiological functions well understood. Here we investigated the expression of JLP in the mouse testis during development. Of the tissues examined, JLP was strongest in the testis, with the most intense staining in the elongated spermatids. Since the anti-JLP antibody used in this study can recognize both JLP and sperm-associated antigen 9 (SPAG9), a splice variant of JLP that has been studied extensively in primates, we also examined its expression in macaque testis samples. Our results indicated that in mouse and primate testis, the isoform expressed at the highest level was JLP, not SPAG9. We also investigated the function of JLP by disrupting the Jlp gene in mice, and found that the male homozygotes were subfertile. Taken together, these observations may suggest that JLP plays an important role in testis during development, especially in the production of functionally normal spermatozoa. © 2008 Springer Science+Business Media B.V

Prevalence of and risk factors for postoperative complications after lower third molar extraction : A multicenter prospective observational study in Japan

Lower third molar extraction is the most common surgical treatment among routine dental and oral surgical procedures. while the surgical procedures for lower third molar extraction are well established, the difficulty of tooth extraction and the frequency of postoperative complications differ depending on the patient’s background. To establish a management protocol for the lower third molars, the prevalence of and risk factors for postoperative complications after lower third molar extraction were investigated in a large number of Japanese patients in a multicenter prospective study. During 6 consecutive months in 2020, 1826 lower third molar extractions were performed at the 20 participating institutions. The medical records of the patients were reviewed, and relevant data were extracted. The prevalence of and risk factors for postoperative complications were analyzed. The prevalence of postoperative complications after lower third molar extraction was 10.0%. Multivariate analysis indicated that age (≤32 vs >32, odds ratio [OR]: 1.428, 95% confidence interval [95% CI]: 1.040–1.962, P < .05), the radiographic anatomical relationship between the tooth roots and mandibular canal (overlapping of the roots and canal vs no close anatomical relationship between the roots and the superior border of the canal, OR: 2.078, 95% CI: 1.333–3.238, P < .01; overlapping of the roots and canal vs roots impinging on the superior border of the canal, OR: 1.599, 95% CI: 1.050–2.435, P < .05), and impaction depth according to the Pell and Gregory classification (position C vs position A, OR: 3.7622, 95% CI: 2.079–6.310, P < .001; position C vs position B, OR: 2.574, 95% CI: 1.574–4.210, P < .001) are significant independent risk factors for postoperative complications after lower third molar extraction. These results suggested that higher age and a deeply impacted tooth might be significant independent risk factors for postoperative complications after lower third molar extraction

HISTOPATHOLOGICAL AND IMMUNOHISTOCHEMICAL ANALYSIS OF ADEN OMATOUS HYPERPLASIA AN D HEPATOCELLULAR CARCINOMA: Cellularity, Thickness of Cell Cord, and Ki-67 Proliferative Activity

To characterize adenomatous hyperplasia (AH) and hepatocellular carcinoma (HCC), and to establish their histopathological differences, morphometrical and immunohistochemical analyses, namely, cellularity, thickness of cell cord, and Ki-67 labeling index (Ki-67 LI) were done on surgically obtained hepatic lesions from patients with positive serum antibody against HCV. The hepatic lesions analyzed include chronic active hepatitis (CAH) (11 specimens), regenerative nodules of liver cirrhosis (LC) (29), AH (11), small HCC Edmondson’s Grade I (GI) (19), GII (26), and GIII (14)The results showed that AH has relatively high cellularity, and significantly greater thickness of Cell cord than LC; whereas, HCC GI has significantly higher cellularity and Ki-67 LI than AH. From the data of these markers, and from the absence of conspicuous structural atypism, AH is considered to be in a different category from HCC GI. The premalignant potential of AH is supported only by its high incidence of coexistence adjacent to HCC GI or GII (6/11). Most lesions of HCC seem to develop from the liver tissue having a background of CAH or LC without passing through AH. Focal fatty changes are frequently observed within lesions of both AH and HCC GI (5/11, 8/19). When non fatty regions of AH and HCC GI are compared, with respect to their markers, particularly Ki-67 LI, as well as the structural atypism, such as microacinus formation and pseudoglandular structure, and invasive growth into the surrounding liver parenchyma, HCC GI can be diagnosed as an early or well-differentiated malignant lesion

Which symptoms negatively affect the oral health-related quality of life in patients with osteonecrosis of the jaw?

Objectives. One of the treatment goals for osteonecrotic lesions of the jaw, such as medication-related osteonecrosis of the jaw (MRONJ) or osteoradionecrosis (ORN), is restoration of quality of life (QOL). This study aimed to identify symptoms that negatively affect QOL in patients with unhealed MRONJ or ORN. Study Design. This cross-sectional study included patients who were previously diagnosed with MRONJ or ORN and who underwent treatment at the Kobe University Hospital between June 2015 and February 2016. Patient QOL was measured by using the Oral Health Impact Profile (OHIP-14). The predictor variable was disease status (stage and healing). The outcome variable was OHIP-14. One-way analysis of variance and Tukey's test were performed. Results. The study included 74 patients (37 men and 37 women; mean age 70 years). Although there was no significant difference between the OHIP-14 scores of unhealed MRONJ and ORN (stages 1-3) and those of healed ones, the "worsened sense of taste" resulted in significant differences among stages in patients with unhealed MRONJ (P = .027) and the "painful mouth aching" in patients with unhealed ORN (P = .041). Conclusions. Worsened sense of taste and pain negatively affected QOL in patients with unhealed MRONJ and ORN