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The replication fork temporarily stalls when encountering an obstacle on the DNA, and replication resumes after the barrier is removed. Simultaneously, activation of the replication checkpoint delays the progression of S phase and inhibits late origin firing. Camptothecin (CPT), a topoisomerase I (Top1) inhibitor, acts as aDNAreplication barrier by inducing the covalent retention of Top1 on DNA. The Timeless-Tipin complex, a component of the replication fork machinery, plays a role in replication checkpoint activation and stabilization of the replication fork. However, the role of the Timeless-Tipin complex in overcoming the CPT-induced replication block remains elusive. Here, we generated viable TIPIN gene knock-out (KO) DT40 cells showing delayed S phase progression and increased cell death. TIPIN KO cells were hypersensitive to CPT. However, homologous recombination and replication checkpoint were activated normally, whereas DNA synthesis activity was markedly decreased in CPT-treated TIPIN KO cells. Proteasome-dependent degradation of chromatin-bound Top1 was induced in TIPINKOcells upon CPT treatment, and pretreatment with aphidicolin, a DNA polymerase inhibitor, suppressed both CPT sensitivity and Top1 degradation. Taken together, our data indicate that replication forks formed without Tipin may collide at a high rate with Top1 retained on DNA by CPT treatment, leading to CPT hypersensitivity and Top1 degradation in TIPIN KO cells. [ABSTRACT FROM AUTHOR]

Hatchery produced leptocephalus larvae of the Japanese eel (Anguilla japonica) were reared on a research vessel and fed natural particulate organic matter (POM) collected in theWestern North Pacific. A small net (36 cm diameter, 60 mm mesh) was vertically towed from 200m depth to surface in 2013, and POM-enriched water (POMEW) filtered through 350 mm mesh was fed to the leptocephali for 11 days. Although the swollen gut corroborated the active ingestion of POM by the leptocephali, low survival rate and heavily melanized gut followed by necrosis in the mid-hindgut region of the leptocephali were observed. A large net (1.14m diameter, 30 mm mesh) was used in 2014 and 2015, which was horizontally drifted subsurface (100-175 m). POMEWs filtered through 53 or 25 mm meshes were fed to the leptocephali for 5-18 days. Neither melanized gut nor necrosis occurred, but considerably low survival rate and little growth comparable with that in a starved condition were observed. No apparent shift in stable carbon and nitrogen isotope ratios was observed in the reared leptocephali. These indicated that the ingested POMs were not assimilated by the leptocephali and suggested that smaller particles may be important for the leptocephali. [ABSTRACT FROM AUTHOR]

Background. Recombinant human erythropoietin (rHuEpo) is a definitive treatment for anaemia in chronic kidney disease (CKD). During long-term rHuEpo treatment most patients develop and show persistent iron deficiency in spite of oral iron supplementation. Abnormalities of iron absorption and transport in the duodenum may contribute to this deficiency. Methods. To investigate changes in iron absorption and transport in CKD and iron deficiency against the background of rHuEpo treatment, we used severely anaemic rats with adenine-induced renal failure (adenine rats) and sham-treated control rats given only the vehicle. After 4 weeks on adenine or the vehicle, the rats were divided into four groups according to whether or not they received rHuEpo for the next 4 weeks: rHuEpo(−)-adenine, rHuEpo(−)-control, rHuEpo(+)-adenine and rHuEpo(+)-control. We evaluated the effects of rHuEpo treatment on iron balance, duodenal mRNA expression of molecules related to iron absorption and transport and hepatic mRNA expression of hepcidin. Results. Treatment with rHuEpo improved anaemia and induced iron deficiency only in the adenine rats, in whom the expression of mRNAs for ferroportin 1 and hephaestin 1 increased and for divalent metal transporter 1 (DMT1) was unchanged. In contrast, control rats treated with rHuEpo showed no changes. Hepcidin mRNA expression was greater in adenine rats than in control rats. Conclusions. In the adenine rats, rHuEpo treatment improved renal anaemia and induced persistent iron deficiency. An alteration of mRNA expression of molecules related to iron metabolism in renal insufficiency may be one of the reasons for this iron deficiency. [ABSTRACT FROM AUTHOR]