Safer Vitrification of Mouse and Human Embryos Using the Novel Cryoroom Vitrification System for Assisted Reproductive Technology.

BACKGROUND: Vitrification is widely used for assisted reproductive technology (ART). Most vitrification devices require the skillful placement of embryos into the carrier and aspiration of excessive vitrification solution. OBJECTIVE: To evaluate the efficacy and safety of the Cryoroom as a vitrification device. MATERIALS AND METHODS: Mouse and human embryos were vitrified with Cryoroom or Cryotop, and the developmental potency was assessed in vitro. Mouse monozygotic twin blastocysts were vitrified with Cryoroom or Cryotop for microarray analysis. RESULTS AND DISCUSSION: In mouse and human embryos, there were no differences between the survival and developmental progress in each device. In silico, the Cryoroom device showed no changes, particularly in DNA methylation after vitrification compared with the Cryotop. These results showed that the form and function of the device may affect the gene expression levels in vitrified embryos. CONCLUSION: The Cryoroom represents a safe and potentially revolutionary vitrification device for ART.

Structures and immunolocalization of Na+, K+ -ATPase, Na+ /H+ exchanger 3 and vacuolar-type H+ -ATPase in the gills of blennies (Teleostei: Blenniidae) inhabiting rocky intertidal areas.

The structure and immunolocalization of the ion transporters Na(+) ,K(+) -ATPase (NKA), Na(+) /H(+) exchanger (NHE3) and vacuolar-type H(+) -ATPase (VHA) were examined in the gills of teleosts of the family Blenniidae, which inhabit rocky shores with vertical zonation in subtropical seas. These features were compared among the following species with different ecologies: the amphibious rockskipper blenny Andamia tetradactylus, the intertidal white-finned blenny Praealticus tanegasimae and the purely marine yaeyama blenny Ecsenius yaeyamaensis. Light and electron microscopic observations indicated that thick gill filaments were arranged close to each other and alternately on two hemibranches of a gill arch in the opercular space of A. tetradactylus. Many mucous cells (MC) and mitochondrion-rich cells (MRC) were present in the interlamellar regions of the gill filament. An immunohistochemical study demonstrated that numerous NKA, NHE3 and some VHA were located predominantly on presumed MRCs of gill filaments and at the base of the lamellae. Analyses using serial (mirror image) sections of the gills indicated that only a few NKA immunoreactive cells (IRC) were colocalized with VHA on some MRCs in the filaments. In the gills of P. tanegasimae, NKA- and NHE3-IRCs were observed in the interlamellar region of the filaments and at the base of the lamellae. VHA-IRCs were located sparsely on the lamellae and filaments. In the gills of E. yaeyamaensis, the lamellae and filaments were thin and straight, respectively. MCs were located at the tip as well as found scattered in the interlamellar region of gill filaments. NKA-, NHE3- and VHA-IRCs were moderately frequently observed in the filaments and rarely on the lamellae. This study shows that the structure and distribution of ion transporters in the gills differ among the three blennid species, presumably reflecting their different ecologies.

Binding analysis of peptides that recognize preferentially cis-azobenzene groups of synthetic polymers.

Peptides identified by the PD method against a film surface composed of azobenzene-containing synthetic polymers were characterized by QCM measurements. Among the four peptides analyzed, the c16 peptide with the sequence Trp-His-Thr-Leu-Pro-Asn-Ala showed the highest binding affinity to the films rich in cis-azobenzene groups. SPR served to determine the association/dissociation constants of the c16 peptide against the trans- and cis-azobenzene groups. The binding constants were estimated to be 1.3 × 10(5) and 1.4 × 10(6) /M, respectively, indicating a high specificity of the c16 peptide for cis-azobenzene conformer. Then mutants of the c16 peptide were synthesized and characterized to gain information on the structural requirements for the cis-form specificity. An Ala-scan clearly revealed that all the amino acid residues of the c16 peptide were essential for the specificity. More importantly, the results with peptides of inverted sequence and containing Gly insertions confirmed that the specificity is strictly derived from the primary sequence of the c16 peptide.

Biological significance of plasminogen activator inhibitor-1 expression in ovarian clear cell adenocarcinoma.

PURPOSE: Ovarian clear cell adenocarcinoma (OCCA) has been reported to display different characteristics from other histological types of epithelial ovarian cancer, and especially differs from serous adenocarcinoma. We investigated plasminogen activator inhibitor-1 (PAI-1) expression in patients with OCCA and attempted to assess its biological significance. METHODS: Fifty-seven patients with OCCA were enrolled. We used formalin-fixed, paraffin-embedded sections of the primary tumor obtained at the first operation to investigate the immunohistochemical expression of PAI-1 and the association of PAI-1 expression with various clinicopathologic factors. RESULTS: The 57 patients were classified into a high PAI-1 expression group and a low expression group. Comparison between the two groups revealed that the percentage of patients with concomitant endometriosis was significantly larger in the high expression group, while the percentage of Stage I patients with positive peritoneal cytology was significantly larger in the low expression group. Comparison of cumulative 5-year survival rates showed that the high expression group had a better prognosis than the low expression group. CONCLUSION: These data suggest an association between concomitant endometriosis and increased expression of PAI-1 in OCCA. The data also suggest that PAI-1 expression influences both peritoneal dissemination of early OCCA and the prognosis.

Cauda Equina and Filum Terminale Arteriovenous Fistulas: Anatomic and Radiographic Features.

Intradural AVF below the conus medullaris may develop either on the filum terminale or the cauda equina (lumbosacral and coccygeal radicular nerves). Although not a few filum terminale AVFs are found in the literature, only 3 detailed cauda equina AVFs have been reported. Here, we analyze the angiographic and MR imaging findings of our cauda equina and filum terminale AVF cases, supplemented with literature research to characterize the radiologic features of the 2 entities. On angiography, filum terminale AVFs were invariably supplied by the extension of the anterior spinal artery accompanied by a closely paralleling filum terminale vein. Cauda equina AVFs were fed by either a radicular or a spinal artery or both arteries, often with a characteristic wavy radicular-perimedullary draining vein. On thin-section axial MR imaging, the filum terminale AVF draining vein joined the spinal cord at the conus medullaris apex, but that of the cauda equina AVF joined above the conus medullaris apex.

Control system for the new RIKEN 28-GHz superconducting electron cyclotron resonance ion source for SRILAC.

A new RIKEN 28-GHz superconducting electron cyclotron resonance ion source (SC-ECRIS) has been installed for the superconducting RIKEN linear accelerator (SRILAC). The new SC-ECRIS control system mainly consists of programmable logic controllers (PLCs) embedded with the Experimental Physics and Industrial Control System. To improve the reliability as compared with previous control systems, two types of PLC central processing units, sequential and Linux, have been installed in the same unit. Past experience has shown that new types of designs that can rapidly respond to system scalability are key. By connecting PLC stations using star-topology field buses, their rapid and cost-effective response to system changes is realized for the new devices. Furthermore, a unique data acquisition system employing a 920-MHz-band radio was developed to measure analog data such as the temperature at the high-voltage stage.

Cardio-cephalic neural crest syndrome: A novel hypothesis of vascular neurocristopathy.

A novel hypothesis proposes that "cardio-cephalic neural crest (NC) syndrome," i.e. cephalic NC including cardiac NC, contributes to the concurrent occurrence of vascular diseases in the cardio- and cerebrovascular regions. NC is a transient structure present in early embryogenesis. Cephalic NC provides mesenchymal cells to the vascular media in these regions. Concurrent cardio- and cerebrovascular lesions have been reported in PHACE syndrome, ACTA2 mutation syndrome, and less frequently in the spontaneous occlusion of the circle of Willis (so-called moyamoya disease). Cardiovascular lesions in these syndromes include coarctation of the aorta, persistent truncus arteriosus, patent ductus arteriosus, and coronary artery disease, and cerebrovascular lesions include agenesis and stenosis/occlusion of the internal carotid arteries, and moyamoya phenomenon. These concurrent vascular lesions both in the cardio- and cerebrovascular regions might be related to cephalic NC. This hypothesis, although not proven, may facilitate a better understanding of the above-mentioned NC-related vascular pathologies and lead to appropriate diagnostic and therapeutic approaches for clinicians and chart future direction for researchers.

Control system renewal for efficient operation in RIKEN 18 GHz electron cyclotron resonance ion source.

A RIKEN 18 GHz electron cyclotron resonance ion source (18 GHz ECRIS) is used as an external ion source at the Radioactive Ion Beam Factory (RIBF) accelerator complex to produce an intense beam of medium-mass heavy ions (e.g., Ca and Ar). In most components that comprise the RIBF, the control systems (CSs) are integrated by the Experimental Physics and Industrial Control System (EPICS). On the other hand, a non-EPICS-based system has hardwired controllers, and it is used in the 18 GHz ECRIS CS as an independent system. In terms of efficient and effective operation, the 18 GHz ECRIS CS as well as the RIBF CS should be renewed using EPICS. Therefore, we constructed an 18 GHz ECRIS CS by using programmable logic controllers with embedded EPICS technology. In the renewed system, an operational log system was developed as a new feature, for supporting of the 18 GHz ECRIS operation.

Progress towards synthetic enzymes for phosphoester hydrolysis.

Synthesis of artificial enzymes for catalyzing phosphoester hydrolysis has been attracting interest for a long time. The remarkable discovery that lanthanide ions catalyze the hydrolysis of DNA and RNA spurred the trend. Currently, progress is being made, mainly in the preparation of homogeneous catalysts, the promotion of catalytic activity by using acid/base cooperation within catalysts, the detailed understanding of the reaction mechanisms involved, and the design of artificial enzymes expressing high specificity and catalytic turn-over.

Second international guidelines for the diagnosis and management of hereditary hemorrhagic telangiectasia

Hereditary hemorrhagic telangiectasia (HHT) is an autosomal dominant disease with an estimated prevalence of 1 in 5000 that is characterized by the presence of vascular malformations (VMs). These result in chronic bleeding, acute hemorrhage, and complications from shunting through VMs. The goal of the Second International HHT Guidelines process was to develop evidence-based consensus guidelines for the management and prevention of HHT-related symptoms and complications. The guidelines were developed using the AGREE II (Appraisal of Guidelines for Research and Evaluation II) framework and GRADE (Grading of Recommendations Assessment, Development and Evaluation) methodology. The guidelines expert panel included expert physicians (clinical and genetic) in HHT from 15 countries, guidelines methodologists, health care workers, health care administrators, patient advocacy representatives, and persons with HHT. During the preconference process, the expert panel generated clinically relevant questions in 6 priority topic areas. A systematic literature search was done in June 2019, and articles meeting a priori criteria were included to generate evidence tables, which were used as the basis for recommendation development. The expert panel subsequently convened during a guidelines conference to conduct a structured consensus process, during which recommendations reaching at least 80% consensus were discussed and approved. The expert panel generated and approved 6 new recommendations for each of the following 6 priority topic areas: epistaxis, gastrointestinal bleeding, anemia and iron deficiency, liver VMs, pediatric care, and pregnancy and delivery (36 total). The recommendations highlight new evidence in existing topics from the first International HHT Guidelines and provide guidance in 3 new areas: anemia, pediatrics, and pregnancy and delivery. These recommendations should facilitate implementation of key components of HHT care into clinical practice.

Sequence-specific and hydrolytic scission of DNA and RNA by lanthanide complex-oligoDNA hybrids.

Totally synthetic and sequence-specific nucleases and ribonucleases, which hydrolyze DNAs and RNAs selectively at target sites, have been prepared. Lanthanide ions, which efficiently hydrolyze the phosphodiester linkages in nucleic acids, are attached to the 5'-end of synthetic DNA oligomers (sequence-recognizing sites) by use of an iminodiacetate ligand. Under physiological conditions, the hybrids selectively hydrolyze substrate DNA or RNA at the 3'-side of the sequence which is complementary with the DNA oligomers in the hybrids. The cerium(IV) ion is the most active as to DNA scission, whereas the europium(III), thulium(III), and lutetium(III) ions are the most effective for RNA scission. All the scissions proceed totally via hydrolysis of the phosphodiester linkages, in the same way as the scissions by natural nucleases and ribonucleases do. The artificial enzymes, which show far greater sequence-specificities than natural ones, exhibit high potential for application to molecular biology, biotechnology, and therapy.

Selective hydrolysis of tRNA by ethylenediamine bound to a DNA oligomer.

A sequence-selective artificial ribonuclease was prepared by attaching ethylenediamine to the 5'-end of a DNA oligomer as the sequence-recognizing moiety. The hybrid, incorporating a 19-mer DNA which is complementary with the A44-A62 sequence of tRNA(Phe), hydrolyzed the tRNA selectively at the 3'-side of C63.

Hydrolysis of DNA and RNA through cooperation of two metal ions: a novel mimic of phosphoesterases.

The phosphodiester linkages in DNA and RNA are efficiently hydrolyzed through cooperation of the lanthanum(III) ion with the iron(III) or tin(IV) ion, although each of these metal ions exhibits poor activity. The conversion on the hydrolysis of adenylyl(3'-5')adenosine by the lanthanum(III)/iron(III) combination (10 mM each) at pH 7.0 and 50 degrees C for 5 min is 72 mol%, whereas the catalysis by the lanthanum or iron ion alone is only marginal. The activity of their combination in the hydrolysis of thymidyl(3'-5')thymidine is more than 15-fold greater than the sum of the activity of each of the two metal ions.

Enormously fast RNA hydrolysis by lanthanide(III) ions under physiological conditions: eminent candidates for novel tools of biotechnology.

Lanthanide(III) ions have shown enormous catalyses for the hydrolysis of the phosphodiester linkages in RNA, indicating their high potential for versatile applications to biotechnology and molecular biology. The activity monotonically increases with increasing atomic number in the lanthanide series, the last three ions (Tm3+, Yb3+, and Lu3+) being the most active. Non-lanthanide metal ions are virtually inactive. The pseudo first-order rate constant for the hydrolysis of adenylyl(3'-5')adenosine (ApA) by LuCl3 (5 mmol x dm(-3)) at pH 7.2 and 30 degrees C is 1.9 x 10(-1) min(-1) (the half-life is only 3.6 min), corresponding to 10(8)-fold acceleration. The product is an equimolar mixture of adenosine and its 2'- or 3'-monophosphate without any byproducts. The 2',3'-cyclic monophosphate of adenosine is not accumulated much in the reaction mixture. Lanthanide ions also efficiently hydrolyze oligoribonucleotides without a specific base-preference. In ApA hydrolysis by NdCl3 and GdCl3, the dependence of the hydrolysis rate on either the pH or concentration of the metal salt coincides fairly well with the corresponding profile of the equilibrium concentration of the bimetallic hydroxo-cluster [M2(OH)2]4+ (M=metal ion). Both the formation of the pentacoordinated intermediate and its decomposition are greatly promoted by lanthanide ions. A catalytic mechanism in which two metal ions (or their coordination water) in these tetracationic hydroxo-clusters show acid/base cooperation is proposed.

Dinuclear lanthanum(III) complex for efficient hydrolysis of RNA.

A dinuclear La3+ complex hydrolyzes the phosphodiester bond in diribonucleotides efficiently under mild conditions. The rate of hydrolysis is remarkably accelerated on the complex formation between La3+ and TPHP (TPHP = N,N,N',N'-tetrakis [(2-pyridyl)methyl]-2-hydroxy-1,3-diaminopropane), and the highest activity is attained when the dinuclear structure is formed. The half-life of ApA hydrolysis by the dinuclear La3+ complex (1 mM) is 350 s, whereas that by the free La3+ ion (1 mM) is 23,000 s at pH 7.2 and 50 degrees C. The dinuclear La3+ complex is promising as a catalytic center of artificial nucleases.

Lanthanide ions for the first non-enzymatic formation of adenosine 3',5'-cyclic monophosphate from adenosine triphosphate under physiological conditions.

Adenosine 3',5'-cyclic monophosphate (cAMP) is formed from adenosine triphosphate at pH 8 and 50 degrees C by use of lanthanide ions. Pr3+ and La3+ are the most active. The cAMP formation is more efficient at higher pH, where the mixture is made homogeneous by the addition of beta-cyclodextrin. The potential functioning of lanthanide ions as the catalytic center of an artificial adenylate cyclase is indicated.

Catalytically active species for CeCl3-induced DNA hydrolysis.

Cerium(III) chloride efficiently hydrolyzes DNA at pH 7 under aerobic conditions. A titration study showed that Ce(III) is oxidized to Ce(IV) in the reaction mixture and the resultant Ce(IV) is responsible for the DNA hydrolysis.

Transient involuntary movement of the leg (monoballismus) during cerebral angiography.

Transient involuntary movement of the leg developed during diagnostic or therapeutic angiography in five patients. The movement was jerky, brisk flexion/extension of the hip and knee joints on the side contralateral to acute ischemia. This movement may occur as a result of thromboembolic cerebral ischemia or as an ischemic complication of angiography.

The accessory middle cerebral artery as a collateral blood supply.

We describe two cases of acute embolic occlusion of the internal carotid artery and the middle cerebral artery in association with a patent accessory middle cerebral artery. Because of the presence of the accessory middle cerebral artery, the frontal lobe was salvaged to some extent, but it did not provide sufficient collateral blood supply to the middle cerebral artery territory to spare the rest of the frontal lobe, the temporal lobe, and the basal ganglia.

Traumatic carotid cavernous sinus fistula: serial angiographic studies from the day of trauma.

BACKGROUND AND PURPOSE: The purpose of this study was to ascertain the early angiographic features characteristic of traumatic carotid cavernous sinus fistulas (CCFs). METHODS: Eight patients with severe craniofacial injuries underwent emergency diagnostic and therapeutic angiography for intractable oronasal bleeding, starting on an average of 6.7 hours after trauma. Carotid angiograms and the clinical manifestation of traumatic CCFs were then reviewed retrospectively to determine characteristic angiographic features. RESULTS: In four of the eight patients, no arteriovenous fistulas were found in the cavernous sinuses and symptomatic CCF did not occur during the follow-up period. In the remaining four patients, dural CCFs (Barrow type B) were observed, unilaterally in three patients and bilaterally in one. One of these four patients subsequently became symptomatic and required transarterial coil embolization. CONCLUSION: Traumatic dural CCFs are frequently observed in the early stage of severe craniofacial trauma, if investigated. Although their spontaneous disappearance is known, some of these do become symptomatic and need treatment.