Spore morphology of Taenitis, Syngramma, and Austrogramme species (Pteridoideae, Pteridaceae) from South-Eastern Asia and Oceania. II

This paper continues consideration of the spores of three paleotropical fern genera – Taenitis, Syngramma, and Austrogramme (Pteridoideae, Pteridaceae) from South-Eastern Asia and Oceania. At the second stage, we carried out a comparative scanning electron microscopy study of spores of three species of Austrogramme, four species of Syngramma, and six species of Taenitis and added information about previously studied spores of seven species of these genera. Spores of all examined species are trilete, tetrahedral or tetrahedral-globose with convex to hemispherical distal side and plane, convex or conical proximal side. The spores of Austrogramme species are the smallest, simplest in ornamentation and similar to each other. Sculpture of the proximal and distal sides are microverrucate, the surface of the spores is covered by granular deposits. Spores of most Syngrammaspecies are very similar to spores of Austrogramme species in shape and surface sculpture: their distal and proximal surfaces are microverrucate, whereas the spores of S. borneensis and S. cartilagidens have the low-tuberculate sculpture. Spores of Taenitis species are very different from the spores of Austrogramme and Syngramma. Seven of nine studied species have spores with well-expressed cingulum (T. blechnoides, T. cordata, T. diversifolia, T. interrupta, T. luzonica, T. obtusa, and T. requiniana), three species (T. cordata, T. hookeri, and T. pinnata) have spores with prominent laesural ridges. The spores have well-expressed ornamentation – tuberculate, baculate, rugate, tuberculate-rugate. The most conspicuous character of the ornamentation of spore surfaces is the presence of rodlets associated with sculpture elements. The densest rodlets are characteristic of Taenitis diversifolia, T. luzonica, T. obtusa, and T. requiniana. Spore size (equatorial diameter) ranges on average between 22 μm and 37 μm in Austrogramme, between 27 μm and 41 μm in Syngramma, and between 26 and 51 μm in Taenitis species.

Comparison of digital and silicone impressions for single-tooth implants and two- and three-unit implants for a free-end edentulous saddle

Background The use of intraoral scanners (IOS) has facilitated the use of digital workflows for the fabrication of implant-supported prostheses not only for single missing teeth, but also for multiple missing teeth. However, the clinical application of IOS and computer-aided design/manufacturing (CAD/CAM) in implant-supported prosthodontics remains unclear. This study aimed to compare the accuracy of digital and silicone impressions for single-tooth implants for bounded edentulous spaces and two-unit and three-unit implant-supported fixed dental prostheses for free-end edentulous spaces. Methods This study enrolled 30 patients (n = 10 for each of the three groups) with an average age of 61.9 years. Conventional silicone-based and digital IOS-based impressions were made for all patients, and the implant superstructures were fabricated. We measured the scan-body misfit and compared the accuracy of the impressions for single-unit, two-unit, and three-unit implant prostheses with a bounded edentulous space by superimposing the standard triangulated language (STL) data obtained from IOS over the STL data of the plaster model used for final prosthesis fabrication. The scan bodies of the superimposed single-molar implant, two-unit implant prosthesis without teeth on the mesial side, two-unit implant prosthesis without teeth on the distal side, three-unit implant prosthesis without teeth on the mesial side, and three-unit implant prosthesis without teeth on the distal side were designated as A, B1, B2, C1, and C2, respectively. The misfit for each scan body was calculated and the accuracies were compared using the Tukey–Kramer method. Results The average scan-body misfit for conditions A, B1, B2, C1, and C2 was 40.5 ± 18.9, 45.4 ± 13.4, 56.5 ± 9.6, 50.7 ± 14.9, and 80.3 ± 12.4 μm, respectively. Significant differences were observed between the accuracies of A and B2, A and C2, and C1 and C2 (P < 0.001). Conclusions IOS and CAD/CAM can find clinical applications for implant-supported prostheses of up to three units for a bounded edentulous saddle. The use of IOS could render implant treatment easier, benefiting both the surgeons and patients. Prosthesis maladjustment may lead to peri-implantitis and prosthetic fracture. Therefore, further validation of the accuracy of IOS impressions is required in patients with multiple missing teeth in long-span implant prostheses.

WDR60-mediated dynein-2 loading into cilia powers retrograde IFT and transition zone crossing

The dynein-2 motor complex drives retrograde intraflagellar transport (IFT), playing a pivotal role in the assembly and functions of cilia. However, the mechanisms that regulate dynein-2 motility remain poorly understood. Here, we identify the Caenorhabditis elegans WDR60 homolog (WDR-60) and dissect the roles of this intermediate chain using genome editing and live imaging of endogenous dynein-2/IFT components. We find that loss of WDR-60 impairs dynein-2 recruitment to cilia and its incorporation onto anterograde IFT trains, reducing the availability of the retrograde motor at the ciliary tip. Consistently, we show that less dynein-2 motors power WDR-60-deficient retrograde IFT trains, which move at reduced velocities and fail to exit cilia, accumulating on the distal side of the transition zone. Remarkably, disrupting the transition zone's NPHP module almost fully restores ciliary exit of underpowered retrograde trains in wdr-60 mutants. This work establishes WDR-60 as a major contributor to IFT and the NPHP module as a roadblock to dynein-2 passage through the transition zone.

THE POTENTIAL OF MICROCOMPUTED TOMOGRAPHY IN STUDYING THE VARIANT MORPHOLOGY OF THE DENTAL CANAL-ROOT SYSTEM

— High-resolution microfocus computed tomography performed on a Skyscan 1176 (Bruker) device was used to study morphological features of the canal-root systems of the upper jaw first molars in residents of the European part of Russia. An analysis of coronal, axial and sagittal sections of 134 upper jaw first permanent molars removed for medical reasons revealed variations in the canal-root system structure. In 85.8% of the cases, the first upper molars had three separate roots: two vestibular and one palatal; in 14.2% of the cases the roots make up different fusions. The palatal root bent in the buccal direction in 52.9% of the cases, while the mesiobuccal root bent in the distal direction in 87.9% of the cases. The greatest variety of curvatures was observed in the distal-buccal root: towards the mesial side — in 28.3% of the cases, another 21.6% of cases being curved towards the distal side. The palatal root in 90.3% of the cases had one canal; 9.7% of the cases featured two canals, while the distal-buccal root in 73.9% of the cases had one canal, and two canals — in 26.1% of the cases. In 89.6% of the cases, the mesiobuccal root had two canals; in 10.4% of the cases — one canal. The mesiobuccal root of the first upper molars was of an oval shape, elongated in the vestibular-oral direction, the most common structures being those of Type IV (43,3%), II (25.4%), III (8.2%) and VI (7.4%) by F. J. Vertucci. The occurrence rate of extra types of root canal structure was 7.4%.

Reproductive biology, embryonic development and matrotrophy in the phylactolaemate bryozoan Plumatella casmiana

Bryozoa is a phylum of aquatic, colonial suspension-feeders within the Lophotrochozoa. In the Phylactolaemata embryonic development occurs in an internal brood sac on the body wall accompanied by extraembryonic nutrition. Owing to previous contradictive descriptions, many aspects of their sexual reproduction require restudy. Consequently, this study analyses embryogenesis of the freshwater bryozoan Plumatella casmiana by serial sections, 3D reconstruction and transmission electron microscopy. Early embryos cleave and soon develop into blastulae with a small central cavity. The mesoderm forms by delamination starting from the distal side towards the proximal end. In later embryos two polypides form on the posterior side that ultimately will be covered by a ciliated mantle in the larva. Embryos increase in size during development and form temporary cell contacts to the embryo sac. Mesodermal cells of the embryo sac show signs of transcellular transport indicating that embryos are nourished by transferring nutrients from the maternal coelom towards the brood cavity. This study clarifies several details such as mesoderm formation and the onset of bud development. Embryos are connected to their respective embryo sacs by a variety of temporary cytoplasmic processes formed by both tissues during embryogenesis, including a ‘placental’ ring zone. Although ultrastructural data of these cell contacts are not entirely conclusive about their function, we suggest that embryos absorb nutrients via the entire surface. The close opposition of embryos to the embryo sac implies placentation as matrotrophic mode in phylactolaemate bryozoans, with embryo sacs acting as placental analogues.

High LET-Like Radiation Tracks at the Distal Side of Accelerated Proton Bragg Peak

Proton therapy is a type of hadron radiotherapy used for treating solid tumors. Unlike heavy charged elements, proton radiation is considered to be low LET (Linear Energy Transfer) radiation, like X-rays. However, the clinical SOBP (Spread Out Bragg Peak) proton radiation is considered to be higher in relative biological effectiveness (RBE) than both X-ray and their own entrance region. The RBE is estimated to be 1.1–1.2, which can be attributed to the higher LET at the SOBP region than at the entrance region. In order to clarify the nature of higher LET near the Bragg peak of proton radiation and its potential cytotoxic effects, we utilized a horizontal irradiation system with CHO cells. Additionally, we examined DNA repair mutants, analyzed cytotoxicity with colony formation, and assessed DNA damage and its repair with γ-H2AX foci assay in a high-resolution microscopic scale analysis along with the Bragg peak. Besides confirming that the most cytotoxic effects occurred at the Bragg peak, extended cytotoxicity was observed a few millimeters after the Bragg peak. γ-H2AX foci numbers reached a maximum at the Bragg peak and reduced dramatically after the Bragg peak. However, in the post-Bragg peak region, particle track-like structures were sporadically observed. This region contains foci that are more difficult to repair. The peak and post-Bragg peak regions contain rare high LET-like radiation tracks and can cause cellular lethality. This may have caused unwanted side effects and complexities of outputs for the proton therapy treatment.

Reduction in organ–organ friction is critical for corolla elongation in morning glory

In complex structures such as flowers, organ–organ interactions are critical for morphogenesis. The corolla plays a central role in attracting pollinators: thus, its proper development is important in nature, agriculture, and horticulture. Although the intraorgan mechanism of corolla development has been studied, the importance of organ–organ interactions during development remains unknown. Here, using corolla mutants of morning glory described approximately 200 years ago, we show that glandular secretory trichomes (GSTs) regulate floral organ interactions needed for corolla morphogenesis. Defects in GST development in perianth organs result in folding of the corolla tube, and release of mechanical stress by sepal removal restores corolla elongation. Computational modeling shows that the folding occurs because of buckling caused by mechanical stress from friction at the distal side of the corolla. Our results suggest a novel function of GSTs in regulating the physical interaction of floral organs for macroscopic morphogenesis of the corolla.

Mycobacterial and Human Ferrous Nitrobindins: Spectroscopic and Reactivity Properties

Structural and functional properties of ferrous Mycobacterium tuberculosis (Mt-Nb) and human (Hs-Nb) nitrobindins (Nbs) were investigated. At pH 7.0 and 25.0 °C, the unliganded Fe(II) species is penta-coordinated and unlike most other hemoproteins no pH-dependence of its coordination was detected over the pH range between 2.2 and 7.0. Further, despite a very open distal side of the heme pocket (as also indicated by the vanishingly small geminate recombination of CO for both Nbs), which exposes the heme pocket to the bulk solvent, their reactivity toward ligands, such as CO and NO, is significantly slower than in most hemoproteins, envisaging either a proximal barrier for ligand binding and/or crowding of H2O molecules in the distal side of the heme pocket which impairs ligand binding to the heme Fe-atom. On the other hand, liganded species display already at pH 7.0 and 25 °C a severe weakening (in the case of CO) and a cleavage (in the case of NO) of the proximal Fe-His bond, suggesting that the ligand-linked movement of the Fe(II) atom onto the heme plane brings about a marked lengthening of the proximal Fe-imidazole bond, eventually leading to its rupture. This structural evidence is accompanied by a marked enhancement of both ligands dissociation rate constants. As a whole, these data clearly indicate that structural–functional relationships in Nbs strongly differ from what observed in mammalian and truncated hemoproteins, suggesting that Nbs play a functional role clearly distinct from other eukaryotic and prokaryotic hemoproteins.

Indocyanine green fluorescence navigation for hepatocellular carcinoma with bile duct tumor thrombus: a case report

Background Bile duct tumor thrombus (BDTT) is one of the features of advanced hepatocellular carcinoma (HCC). In the resection of HCC with BDTT, it is important to detect the BDTT tip to decide the appropriate point of bile duct division. In this regard, the efficacy of indocyanine green (ICG) fluorescence navigation has been confirmed for the detection of HCC, whereas its utility for BDTT has not yet been reported. Herein, we describe our experience with right hepatectomy for HCC with BDTT using ICG fluorescence navigation. Case presentation A 72-year-old woman had experienced local recurrences of HCC after radiofrequency ablation, with BDTT reaching the confluence of the right anterior branch and posterior branch. Right hepatectomy was planned, and 2.5 mg of ICG was injected one day before surgery. After transection of the liver parenchyma, the right liver was connected with only the right hepatic duct. ICG fluorescence imaging visualized the tip of BDTT in the bile duct with clear contrast; the proximal side (hepatic side) of the right hepatic duct showed stronger fluorescence than the distal side (duodenal side). The bile duct was divided at the distal side of the BDTT border, and the tip of BDTT was recognized into the resected right hepatic duct without laceration. The patient had an uneventful postoperative course and currently lives without recurrences for 6 months. Conclusions ICG fluorescence navigation assisted in the precise resection of the bile duct in HCC with BDTT.

An electron transfer competent structural ensemble of membrane-bound cytochrome P450 1A1 and cytochrome P450 oxidoreductase

Cytochrome P450 (CYP) heme monooxygenases require two electrons for their catalytic cycle. For mammalian microsomal CYPs, key enzymes for xenobiotic metabolism and steroidogenesis and important drug targets and biocatalysts, the electrons are transferred by NADPH-cytochrome P450 oxidoreductase (CPR). No structure of a mammalian CYP–CPR complex has been solved experimentally, hindering understanding of the determinants of electron transfer (ET), which is often rate-limiting for CYP reactions. Here, we investigated the interactions between membrane-bound CYP 1A1, an antitumor drug target, and CPR by a multiresolution computational approach. We find that upon binding to CPR, the CYP 1A1 catalytic domain becomes less embedded in the membrane and reorients, indicating that CPR may affect ligand passage to the CYP active site. Despite the constraints imposed by membrane binding, we identify several arrangements of CPR around CYP 1A1 that are compatible with ET. In the complexes, the interactions of the CPR FMN domain with the proximal side of CYP 1A1 are supplemented by more transient interactions of the CPR NADP domain with the distal side of CYP 1A1. Computed ET rates and pathways agree well with available experimental data and suggest why the CYP–CPR ET rates are low compared to those of soluble bacterial CYPs.