r-Process elements from magnetorotational hypernovae.

Neutron-star mergers were recently confirmed as sites of rapid-neutron-capture (r-process) nucleosynthesis1-3. However, in Galactic chemical evolution models, neutron-star mergers alone cannot reproduce the observed element abundance patterns of extremely metal-poor stars, which indicates the existence of other sites of r-process nucleosynthesis4-6. These sites may be investigated by studying the element abundance patterns of chemically primitive stars in the halo of the Milky Way, because these objects retain the nucleosynthetic signatures of the earliest generation of stars7-13. Here we report the element abundance pattern of the extremely metal-poor star SMSS J200322.54-114203.3. We observe a large enhancement in r-process elements, with very low overall metallicity. The element abundance pattern is well matched by the yields of a single 25-solar-mass magnetorotational hypernova. Such a hypernova could produce not only the r-process elements, but also light elements during stellar evolution, and iron-peak elements during explosive nuclear burning. Hypernovae are often associated with long-duration γ-ray bursts in the nearby Universe8. This connection indicates that similar explosions of fast-spinning strongly magnetized stars occurred during the earliest epochs of star formation in our Galaxy.

Extremely metal-poor stars from the cosmic dawn in the bulge of the Milky Way.

The first stars are predicted to have formed within 200 million years after the Big Bang, initiating the cosmic dawn. A true first star has not yet been discovered, although stars with tiny amounts of elements heavier than helium ('metals') have been found in the outer regions ('halo') of the Milky Way. The first stars and their immediate successors should, however, preferentially be found today in the central regions ('bulges') of galaxies, because they formed in the largest over-densities that grew gravitationally with time. The Milky Way bulge underwent a rapid chemical enrichment during the first 1-2 billion years, leading to a dearth of early, metal-poor stars. Here we report observations of extremely metal-poor stars in the Milky Way bulge, including one star with an iron abundance about 10,000 times lower than the solar value without noticeable carbon enhancement. We confirm that most of the metal-poor bulge stars are on tight orbits around the Galactic Centre, rather than being halo stars passing through the bulge, as expected for stars formed at redshifts greater than 15. Their chemical compositions are in general similar to typical halo stars of the same metallicity although intriguing differences exist, including lower abundances of carbon.

The effect of endodontic procedures on apical crack initiation and propagation ex vivo.

AIM: To evaluate the potential effects of endodontic procedures (instrumentation and filling) on crack initiation and propagation in apical dentine. METHODOLOGY: Forty extracted single-rooted premolars with two canals were selected, 1.5 mm of the apex was ground perpendicular to the long axis of the tooth and the surface polished. The specimens were divided into 4 groups. The buccal canals of groups A, B and C were enlarged to size 40 with manual K-files. Group A was filled with gutta-percha using lateral condensation and vertical compaction without sealer. Group B was filled with the same method as group A except only lateral condensation was used. Group C was left unfilled, while group D was left unprepared and unfilled. Images of the resected surface were taken after resection (baseline), after canal preparation, after filling and after 4-week storage. The images were then inspected for cracks originating from the canal. RESULTS: A significant effect of preparation on crack initiation (P < 0.05) and no significant effect of filling (P > 0.05) or 4-week storage on crack initiation (P > 0.05) was found (logistic regression). Fisher's exact test revealed a significant effect of filling on crack propagation (P < 0.05) and no effect of 4-week storage on crack propagation (P > 0.05). CONCLUSIONS: Root canal procedures can potentially initiate and propagate cracks from within the root canal in the apical region.

Investigation of cynomolgus monkey complement.

BACKGROUND: The cynomolgus monkey is one of the most popular recipient animals in xenotransplantation experiments. However, studies of the cynomolgus monkey complement are rare. In the present study, based on the study that compared the hemolytic complement titer in cynomolgus monkeys with that in humans, the complement regulatory function of human decay accelerating factor (CD55) in both human and cynomolgus monkey sera was studied. METHODS: Hemolytic complement titers in cynomolgus monkeys were calculated using the same methods as are used in humans. Next, the complement regulatory function of human DAF (CD55) in cynomolgus monkey serum was studied using porcine endothelial cells (PECs) and human DAF. RESULTS: Of the complement titers tested, such as CH50, ACH50, C4, C2, and C3, the values were relatively high, except for the C4 titer. Human DAF on the surface of PEC resulted in nearly identical complement regulatory function in the human and cynomolgus monkey sera. CONCLUSIONS: Human DAF showed nearly the same complement regulatory function in both human and cynomolgus monkey sera.

An uncommon cleft subtype of unilateral cleft lip and palate.

The finding that the vomer plays a crucial role in maxillary growth suggests that the bilateral cleft configuration of unilateral cleft lip and palate (UCLP), in which the vomer is detached from the non-cleft-side secondary hard palate, negatively influences palatal development, and this hypothesis was tested. Sixty persons with complete UCLP, including those with the vomer detached from (n = 30, b-UCLP) and attached to (n = 30, u-UCLP) the secondary hard palate, were analyzed morphologically, with the use of cast models taken at 10 days, 3 mos, and 12 mos of age. The anterio-posterior palatal length at 12 mos of age in those with b-UCLP was significantly shorter than that in those with u-UCLP, by 8.7% (p < 0.05). In addition, palatal width development in the first year in those with b-UCLP was also significantly retarded. These results suggest that the uncommon bilateral cleft subtype in UCLP should be included in the cleft classification.

Identification of the novel AML1 fusion partner gene, LAF4, a fusion partner of MLL, in childhood T-cell acute lymphoblastic leukemia with t(2;21)(q11;q22) by bubble PCR method for cDNA.

The AML1 gene is frequently rearranged by chromosomal translocations in acute leukemia. We identified that the LAF4 gene on 2q11.2-12 was fused to the AML1 gene on 21q22 in a pediatric patient having T-cell acute lymphoblastic leukemia (T-ALL) with t(2;21)(q11;q22) using the bubble PCR method for cDNA. The genomic break points were within intron 7 of AML1 and of LAF4, resulting in the in-frame fusion of exon 7 of AML1 and exon 8 of LAF4. The LAF4 gene is a member of the AF4/FMR2 family and was previously identified as a fusion partner of MLL in B-precursor ALL with t(2;11)(q11;q23), although AML1-LAF4 was in T-ALL. LAF4 is the first gene fused with both AML1 and MLL in acute leukemia. Almost all AML1 translocations except for TEL-AML1 are associated with myeloid leukemia; however, AML1-LAF4 was associated with T-ALL as well as AML1-FGA7 in t(4;21)(q28;q22). These findings provide new insight into the common mechanism of AML1 and MLL fusion proteins in the pathogenesis of ALL. Furthermore, we successfully applied bubble PCR to clone the novel AML1-LAF4 fusion transcript. Bubble PCR is a powerful tool for detecting unknown fusion transcripts as well as genomic fusion points.

Induction of heme oxygenase-1 by cobalt protoporphyrin enhances the antitumour effect of bortezomib in adult T-cell leukaemia cells.

Adult T-cell leukaemia (ATL) is a lethal neoplasia derived from HTLV-1-infected T lymphocytes frequently exhibiting nuclear factor-kappaB (NF-kappaB) activation. Despite the use of various treatment regimens, the prognosis of ATL is poor, and new treatment strategies are urgently needed. We therefore explored the effect and the molecular mechanism of a proteasome inhibitor, bortezomib, in ATL cells. We found bortezomib-induced cell death, and bortezomib suppressed constitutive NF-kappaB activation via I-kappaB stabilisation in three ATL cell lines (TaY, MT-2 and MT-4). An oligonucleotide DNA microarray analysis of TaY cells revealed upregulation of genes encoding heat shock proteins (HSPA1A, STIP1, HSPA1B, and HSPCA), genes related to protein folding (CDC37 and ANAPC5), Fas-associated factor 1(FAF1) and an oxidative stress-related gene, heme oxygenase-1(HMOX-1), known to be a target gene of hypoxia-inducible gene-1 alpha (HIF-1 alpha). Cobalt protoporphyrin induced HMOX-1, instead of HIF-1 alpha expression and increased bortezomib-induced apoptosis in the presence of pharmacologically effective doses of bortezomib. In contrast, zinc protoporphyrin downregulated HMOX-1 expression, thereby partially inhibiting bortezomib-induced cell death. This indicates that HMOX-1 may modulate anticancer effects of bortezomib in ATL cells, and could be a molecular target in treating ATL patients.

Deletion of entire HLA-A gene accompanied by an insertion of a retrotransposon.

Unusual HLA-A'null' alleles because of an entire gene deletion were found in three apparently unrelated Japanese families with leukemia patients. Inclusion of the entire HLA-A gene in the deletion was confirmed by polymerase chain reaction direct sequencing of the surrounding regions of HLA-A. Further localization of the breakpoints of the HLA-A deletion at the centromeric and telomeric sides was performed, and these families were shown to possess the identical deletion. We then determined the genomic sequence of the HLA-A-deleted haplotype. Surprisingly, the haplotype turned out to carry an insertion of an SVA (SINE-VNTR-Alu) retrotransposon of 2 kb as well as the 14 kb deletion that included the entire HLA-A gene.

Similar acute molecular responses to equivalent volumes of isometric, lengthening, or shortening mode resistance exercise.

The present study was undertaken to test the hypothesis that the contraction mode of action [static-isometric (Iso), shortening-concentric (Con), or lengthening-eccentric (Ecc)] used to stress the muscle provides a differential mechanical stimulus eliciting greater or lesser degrees of anabolic response at the initiation of a resistance training program. We performed an acute resistance training study in which different groups of rodents completed four training sessions in either the Iso, Con, or Ecc mode of contraction under conditions of activation and movement specifically designed to elicit equivalent volumes of force accumulation. The results of this experiment indicate that the three modes of contraction produced nearly identical cell signaling, indicative of an anabolic response involving factors such as increased levels of mRNA for IGF-I, procollagen III alpha1, decreased myostatin mRNA, and increased total RNA concentration. The resulting profiles collectively provide evidence that pure mode of muscle action, in and of itself, does not appear to be a primary variable in determining the efficacy of increased loading paradigms with regard to the initiation of selected muscle anabolic responses.

The effects of file size, sodium hypochlorite and blood on the accuracy of Root ZX apex locator in enlarged root canals: an in vitro study.

BACKGROUND: The initial electronic apex locator (EAL) length measurement is generally established with a small-sized file. It is not known whether file size would be interfering with the reading accuracy of the EAL. This study aimed to evaluate the effect of file size on the accuracy of Root ZX apex locator using an agar model when sodium hypochlorite solution or blood was present during electronic measurements in enlarged root canals. METHODS: A total of 36 extracted lower premolars were used. In stage 1, the canals were instrumented using size 10-40 K-files with a size 40 K-file as the master apical file (MAF). The teeth were then divided randomly into two groups of 18 teeth each. In group A, the teeth were mounted in one per cent agar and irrigated with six per cent sodium hypochlorite solution (NaOCl), while in group B the teeth were mounted in agar and irrigated with human blood. In stage 2, the canals were enlarged using a size 60 K-file as the MAF. In stages 1 and 2, the apical portions of the canals were instrumented using the step-back sequence (up to a size 80 K-file). In stage 3, the canals were enlarged using a size 80 K-file as the MAF. In each stage, the length was measured with a Root ZX until the meter value reached 'APEX' using small and large size files. RESULTS: Three-way ANOVA and Bonferroni test showed that file size, stage of preparation and type of irrigant all had a significant influence on the measurement error (P < 0.0001), with all the interactions between these three factors being significant (P < 0.0001). CONCLUSIONS: As the diameter of the root canal increased, the measured length with the smaller size files became shorter. A file of a size close to the prepared canal diameter should be used for root length measurement in the presence of blood. In the presence of NaOCl, the Root ZX was highly accurate even when the file was much smaller than the diameter of the canal. The agar model was effective and suitable for testing EALs in vitro.

An ex vivo evaluation of a new root canal irrigation technique with intracanal aspiration.

AIM: To evaluate the effectiveness of a new root canal irrigation technique with intracanal aspiration in removing the smear layer and to assess irrigant extrusion ex vivo. METHODOLOGY: Thirty-five instrumented canals of extracted human canine teeth that had been resected apically by removing 3 mm of the root tip were divided into one control and four experimental groups of seven teeth each. The roots were fixed in a plastic case and surrounded with normal saline agar coloured with 1% acid red. No irrigation was performed in the control teeth. Each root canal in the experimental groups was irrigated with 9 mL of 14% ethylenediaminetetraacetic acid for 3 min, and then with 6 mL of 6% sodium hypochlorite (NaOCl) for 2 min. In the intracanal aspiration technique, the irrigant was delivered from the tip of an injection needle placed 12 mm from the apical root-end and an aspiration needle that was connected to a Root ZX apex locator placed 2 and 3 mm short of the apical root-end in groups 1 and 2, respectively. In the conventional method, the tip of an injection needle used for delivery of the irrigant and as an active electrode was placed 2 and 3 mm short of the apical root-end in groups 3 and 4, respectively, the tip of the aspiration needle was placed 12 mm from the apical root-end in these groups. The readings of the Root ZX during irrigation were recorded. The cleanliness of the canal was evaluated by scoring smear layer from scanning electron microscopy (SEM) images of the canal. Extrusion of NaOCl was detected by measuring the discoloured area of the agar around the apical root-end. The data obtained were statistically analysed by one-way anova, the Kruskal-Wallis test and Friedman's test. RESULTS: In the SEM study, the canals in groups 1-3 were significantly cleaner than those in the control and group 4 (P < 0.05). The mean Root ZX readings in groups 1-3 were approximately "0.5". The discoloured area in group 3 was significantly larger than the other groups (P < 0.05). CONCLUSIONS: Irrigation using the intracanal aspiration technique allowed more effective removal of the smear layer than that performed by the conventional method in an apically resected canine tooth. The intracanal aspiration technique produced limited extrusion of the irrigant beyond the apical foramen.

Selective and efficient culturing of retinal pigment epithelial cells using a feeder layer.

BACKGROUND: The techniques to isolate and purify retinal pigment epithelial (RPE) cells from small piece of autologous tissues are extremely difficult, and it is important to develop an efficient cell culture technique for RPE cells. The purpose of this study was to investigate the effect of 3T3-J2 cells and conditioned medium from 3T3-J2 cells on the proliferation of cultured RPE cells. METHODS: RPE cells from pigmented rabbits and a human RPE-derived cell line, ARPE-19, were used. First, the effects of co-culturing RPE cells with 3T3-J2 cells on the growth of the cells were analyzed. Second, the effects of the conditioned medium from 3T3-J2 cells on the proliferation of both types of cells were investigated. And third, the effects of the conditioned medium on RPE cell culture from a surgically removed choroidal neovascular (CNV) membrane were investigated. RESULTS: The 3T3-J2 cells increased the proliferation of both rabbit RPE cells and ARPE-19 cells. The number of rabbit RPE cells cultured in a mixture of the conditioned medium from 3T3-J2 cells was significantly higher than that in the reported optimal condition, and a similar tendency was observed for ARPE-19 cells. The results from enzyme-linked immunosorbent assay showed the presence of PDGF-AB, VEGF and IGF-I in the conditioned medium. The conditioned medium also promoted selective growth of human RPE cells from CNV. DISCUSSION: The results from this study present the conditions for efficient and selective culture of primary RPE cells.

Detection of the second mesiobuccal canal in mesiobuccal roots of maxillary molar teeth ex vivo.

AIM: To assess the effectiveness of magnification and dentine removal (troughing) when locating the second mesiobuccal canal in mesiobuccal roots of maxillary molars. METHODOLOGY: A total of 208 extracted human maxillary molars were examined. After crown and pulp removal, the MB1 and 2 canals in the mesiobuccal root were located in three stages that were performed by two undergraduate dental students. Stage 1: canals were located with an endodontic explorer; stage 2: additional canals in the same teeth were located under magnification with a digital microscope (VH-8000, Keyence, Japan); stage 3: additional canals in the same teeth were located by removing dentine (troughing) from the pulp chamber floor within 3 mm from MB1 canal towards the palatal canal with an Enac ultrasonic tip (ST 21, Osada, Japan). In each group, the canals were prepared with Gates Glidden burs and K-files. The distal and palatal roots were then removed, and Indian ink was injected into the canal system within the mesio-buccal root. The root surfaces were washed with 6% NaOCl, and then rendered transparent to observe canal morphology. The root canal configurations were classified into five categories following the modified Weine's classification. RESULTS: More than one canal in the mesio-buccal root was observed in 48% of specimens. Detection rates of multiple canals were 7, 18 and 42% following stages 1, 2 and 3, respectively. There was a significant difference between the stages for detecting the MB2 canal (P < 0.05, Friedman test). CONCLUSIONS: Both magnification (stage 2) and dentine removal under magnification (stage 3) were effective in detecting the presence of the MB2 canal. However, MB2 canals could not be detected in 13% of the teeth because of canal calcification or branching located more apically.

Analysis of forces developed during mechanical preparation of extracted teeth using RaCe rotary instruments and ProFiles.

AIM: To compare the torque and load during instrumentation with ProFile and RaCe nickel-titanium rotary instruments. METHODOLOGY: Thirty human incisor roots 12 mm in length were embedded in epoxy resin, and divided into three groups of 10 specimens each. Instruments employed in each group were as follows: RaCe .02 Step Back Files in group 1; RaCe .04 Step Back Files in group 2; and .04 ProFiles in group 3. Two load cells, whose outputs were connected to a digital oscilloscope, recorded torque and vertical load. Data were analysed by one-way anova. RESULTS: Torque values were statistically different between groups 1 and 3, and between groups 2 and 3 (torque was higher in group 3 than in groups 1 and 2). Vertical loads were statistically higher in group 3 than in groups 1 and 2. CONCLUSIONS: Torsional and vertical forces can be evaluated during instrumentation of straight root canals using the device and methods described in this study. When the step-back technique was employed, torque and vertical load obtained with RaCe instruments were lower than that obtained with ProFiles.

Evaluation of root-end cavity preparation using ultrasonic retrotips.

AIM: To evaluate and compare the efficiency of root-end preparations using ultrasonic retrotips coated with diamond and zirconium nitride. METHODOLOGY: Eighty-five extracted single-rooted teeth were root filled, and then resected 3 mm from their apices. Root-end cavities were prepared with KiS (zirconium nitride-coated retrotip), CT-5 (stainless steel tip) or diamond-coated (DC) ultrasonic retrotips, and 10 teeth served as controls. Thirty teeth were used for evaluation of the time required to prepare the root-end cavity, the number of microcracks produced on the resected surface and the number of dentinal tubule openings on the root-canal wall using scanning electron microscope (SEM) images. A further 55 teeth were used for evaluation of dye penetration following filling of the root-end cavities with Super EBA. The degree of dye penetration in millimetres was measured under the microscope after 7 days of immersion in India ink. Statistical analyses were performed using the one-way ANOVA and Scheffe's F-test as the post hoc test. RESULTS: There was no significant difference in the number of microcracks and dentinal tubule openings present in the root apices prepared by the three retrotips. The time required for root-end cavity preparation using the DC retrotip was significantly less than that using the other groups (P<0.01). Positive controls showed dye penetration throughout the length of the root-end cavity, and negative controls showed no dye penetration. There was no significant difference between the three experimental groups in dye penetration. CONCLUSIONS: In this laboratory study, the time required to prepare root-end cavities using KiS retrotips was the same as that using CT-5 retrotips, and longer than that using DC retrotips. There was no significant difference in the number of microcracks or dye penetration between the three kinds of retrotips.

Achieving recommended goals for blood-glucose and blood-pressure control in diabetic patients: a multi-center cross-sectional evaluation in the Niigata prefecture.

Hypertension in patients with diabetes mellitus can contribute to a deterioration of the patient's status as much as most diabetes mellitus-related complications. Previous interventional trials have found that to prevent the onset and progression of diabetic microangiopathy, the optimal glycemic threshold is an HbA1c of less than 6.5% and the optimal blood pressure is less than 130/85 mmHg. The present study investigated the prevalence of achieving these recommended goals for glycemic and blood pressure control in type 2 diabetic patients in the Niigata prefecture. A questionnaire was administered in this multi-center study to 3573 patients from 92 participating Niigata Diabetic Complication Study hospitals. Patients aged 63.5 +/- 11 years with type 2 diabetes were evaluated. Only 46.8% of patients achieved the recommended glycemic control level, and only 41.4% of patients achieved the BP target. Moreover, only 20.5% of all patients achieved both target levels for blood glucose and blood pressure. This demonstrates the difficulty in clinical practice of achieving concurrent glucose and blood pressure control in diabetic patients.

Intercalary regeneration in planarians.

How can a planarian regenerate its entire body from a small portion of its body? Neoblasts, the totipotent stem cells of planarian, are assumed to be able to produce all missing cell types. However, we do not know how the cell fate of these cells is controlled during regeneration. Our recent studies with molecular markers suggest that intercalary regeneration is the fundamental principle in planarian regeneration. Here, we introduce the intercalation induced by ectopic grafting along the anteroposterior (A-P), dorsoventral (D-V), and left-right (L-R) axes. Blastema formation is evoked by ectopic D-V interactions after wound closure. Intercalation between the blastema and stump induces rearrangement of the positional identities along the A-P axis. Consequently, totipotent stem cells change their differentiation patterns according to the newly rearranged positional identities along the A-P, D-V, and L-R axes. According to the classic view, the blastema is regarded as the place where undifferentiated cells accumulate and regenerative events occur. Here, we propose a new interpretation, i.e., that the blastema may work as a signaling center inducing intercalary regeneration. Also, the roles of molecules and genes involved in intercalary regeneration are discussed.

A scanning electron microscopic study of dentinal erosion by final irrigation with EDTA and NaOCl solutions.

AIM: The purpose of this in vitro study was to examine dentinal erosion caused by final irrigation with EDTA and NaOCl. METHODOLOGY: Twenty-five single-rooted human teeth were instrumented with rotary nickel-titanium Series 29 Profile Instruments. The teeth were divided into five groups and subjected to final irrigation as follows: group A, irrigated with 6% NaOCl (3 mL) for 2 min; group B, 15% EDTA (3 mL) for 1 min; group C, 15% EDTA (3 mL) for 1 min, followed by 6% NaOCl (3 mL) for 2 min; group D, 15% EDTA (3 mL) for 3 min and group E, 15% EDTA (3 mL) for 3 min, followed by 6% NaOCl (3 mL) for 2 min. Photomicrographs of dentinal walls were produced using a scanning electron microscope (3000 x) at 1, 3 and 6 mm from the apex. The amount of debris and dentinal tubule diameter were evaluated, and values were statistically analysed using one-way ANOVA and Fisher's PLSD test. RESULTS: When the root canal was irrigated with 15% EDTA alone, the dentine had a smooth and plane appearance, and dentinal tubule orifices were regular and separated. When the root canal was irrigated with EDTA followed by NaOCL the dentine was eroded and the dentinal tubule orifices were irregular and rough. Dentinal tubule diameter increased to 3.43 +/- 0.23 microm in group C and to 3.93 +/- 0.44 microm in group E. Significant differences were observed between groups B and C, and between groups D and E (P < 0.05). However, more debris was removed by irrigation with EDTA followed by NaOCl than with EDTA alone (P < 0.05). CONCLUSIONS: Final irrigation with 6% NaOCl accelerates dentinal erosion following treatment with 15% EDTA.