Geographical distribution and phoretic associations of the viviparous nematode Tokorhabditis atripennis with Onthophagus dung beetles in Japan.

Viviparity is generally considered to be rare in animals. In nematodes, only six species of Rhabditida are viviparous. Five of these species have been identified in association with Onthophagus dung beetles, with Tokorhabditis atripennis being repeatedly isolated from the dung beetle Onthophagus atripennis in Japan. T. atripennis is easy to culture in a laboratory setting, and its host, O. atripennis, is distributed all over Japan. Therefore, T. atripennis is an ideal candidate for ecological and evolutionary studies on viviparity. However, the extent of their distribution and relationship with dung beetles, as well as habitats, remain unclear. In the present study, we conducted field surveys and successfully isolated 27 strains of viviparous nematodes associated with tunneler dung beetles from various regions of Japan, all of which were identified as T. atripennis. T. atripennis exhibited a strong association with Onthophagus dung beetles, especially O. apicetinctus and O. atripennis. And it was predominantly found in specific anatomical locations on the beetle bodies, such as the 'groove between pronotum and elytron' and the 'back of the wings'. Our findings suggest that Onthophagus species are the primary hosts for T. atripennis, and T. atripennis exhibits a close relationship with the living environments of tunneler beetles. This association may play a significant role in the evolution of viviparity in nematodes.

Morphological and molecular characterization of Travassosinema viatorum n. sp. (Nematoda: Oxyuridomorpha: Travassosinematidae) from the invasive millipede Chamberlinius hualienensis Wang, 1956 (Diplopoda: Polydesmida: Paradoxosomatidae) in Okinawa, Japan.

Travassosinema viatorum n. sp. (Nematoda: Oxyuridomorpha: Travassosinematidae) is described from the invasive millipede Chamberlinius hualienensis Wang, 1956 (Diplopoda: Polydesmida: Paradoxosomatidae) in Okinawa, Japan. Females of T. viatorum n. sp. resemble T. claudiae Morffe & Hasegawa, 2017; T. thyropygi Hunt, 1996 and T. travassosi Rao, 1958 by lacking both lateral alae and a contraction posterior to the vulva. The body length of T. viatorum n. sp. is shorter than the aforementioned species. It differs from T. claudiae and T. thyropygi by its comparatively longer oesophagus, more posterior vulva, lack of cuticular ribs in the alae of the umbraculum and in the position of the nerve ring and excretory pore. Travassosinema viatorum n. sp. can be differentiated from T. travassosi by the extension of the cephalic umbraculum and the smaller eggs. The phylogeny of T. viatorum n. sp. is inferred by the D2-D3 domains of the 28S rDNA. The new species is located in a monophyletic clade with other sequences of Travassosinema from millipedes. The comparison of the phylogenies of Travassosinema and their hosts points to host switching rather than coevolution as a mechanism to explain the host-nematode relationships. This constitutes the second species of the genus Travassosinema described from the Japanese archipelago and the first oxyuridomorph nematodes found in C. hualienensis.

Medication Adherence of People Living with HIV in Japan-A Cross-Sectional Study.

Long-term medical care for people living with HIV (PLHIV) is critical for treatment efficacy, and various studies have examined reasons for antiretroviral therapy (ART) non-adherence. In Japan, doctors assume patients maintain high adherence. However, little is known about real-world treatment adherence. We conducted an anonymous self-administered web-based survey asking about adherence for a total of 1030 Japanese PLHIV who were currently on ART. Adherence was determined using the eight-item Morisky Medication Adherence Scale (MMAS-8), for which scoring ranged from 0 to 8 and scores < 6 points were classified as low adherence. Data were analyzed based on patient-related factors; therapy-related factors; condition-related factors, such as a comorbidity with depression (utilizing the Patient Health Questionnaire 9, PHQ-9); and healthcare/system-related factors. Among 821 PLHIV who responded to the survey, 291 responders (35%) were identified as being in the low adherence group. A statistically significant relationship was found between the number of missed anti-HIV drug doses within the previous 2 weeks and long-term adherence, per the MMAS-8 score (p < 0.001). Risk factors for low adherence included age (younger than 21 years, p = 0.001), moderate to severe depression (p = 0.002, using the PHQ-9), and drug dependence (p = 0.043). Adherence was also influenced by a shared decision-making process, including treatment selection, doctor-patient relations, and treatment satisfaction. Adherence was mainly affected by treatment decision factors. Hence, support of care providers should be considered critical for improving adherence.

Protocol for cell-type-specific tissue reconstruction in the murine lung fibrogenic microenvironment.

Pulmonary fibrosis is a process characterized by epithelial injury and fibroblast activation. It is also well recognized as a predisposition to lung cancer. Here, we present a protocol to establish an in vivo model to evaluate the dynamics of alveolar epithelial type 2 cells and lung cancer cells in the context of the lung fibrogenic microenvironment. Utilizing the cell transfer technique, we detail a basis for therapeutic approaches in pulmonary fibrosis and tools for precision medicine against lung cancer. For complete details on the use and execution of this protocol, please refer to Miyata et al. (2022).1.

Morphological and molecular characterization of Desmicola ryukyuensis n. sp. (Nematoda: Oxyuridomorpha: Thelastomatidae) from the wood-feeding cockroach Panesthia angustipennis yayeyamensis Asahina, 1988 (Blattaria: Blaberidae) in the Ryukyu Archipelago, Japan.

Desmicola ryukyuensis n. sp. (Nematoda: Oxyuridomorpha: Thelastomatidae) is described from the wood-feeding cockroach Panesthia angustipennis yayeyamensis Asahina, 1988 (Blattaria: Blaberidae) in Iriomote Island, Japan. The males of D. ryukyuensis n. sp. are similar to D. lamdongensis Sokolova, 2019 but differ by the length of the spicule and the extension of the lateral alae. The females of D. ryukyuensis n. sp. resemble those of D. ornata Jex, Schneider, Rose & Cribb, 2005, but can be differentiated by the shape of the sensilla in the interlabial space and the presence of lateral alae in D. ryukyuensis n. sp. that are absent in D. ornata. The females of D. ryukyuensis n. sp. are similar to D. lamdongensis. However, they differ in the morphology of the lips and the size of the eggs. The phylogeny of D. ryukyuensis n. sp. is inferred by the D2-D3 domains of the 28S rDNA. The new species forms a clade with another sequence of an unidentified Desmicola species from a Vietnamese wood-feeding cockroach.

N-WASP-Arp2/3 signaling controls multiple steps of dendrite maturation in Purkinje cells in vivo.

During neural development, the actin filament network must be precisely regulated to form elaborate neurite structures. N-WASP tightly controls actin polymerization dynamics by activating an actin nucleator Arp2/3. However, the importance of N-WASP-Arp2/3 signaling in the assembly of neurite architecture in vivo has not been clarified. Here, we demonstrate that N-WASP-Arp2/3 signaling plays a crucial role in the maturation of cerebellar Purkinje cell (PC) dendrites in vivo in mice. N-WASP was expressed and activated in developing PCs. Inhibition of Arp2/3 and N-WASP from the beginning of dendrite formation severely disrupted the establishment of a single stem dendrite, which is a characteristic basic structure of PC dendrites. Inhibition of Arp2/3 after stem dendrite formation resulted in hypoplasia of the PC dendritic tree. Cdc42, an upstream activator of N-WASP, is required for N-WASP-Arp2/3 signaling-mediated PC dendrite maturation. In addition, overactivation of N-WASP is also detrimental to dendrite formation in PCs. These findings reveal that proper activation of N-WASP-Arp2/3 signaling is crucial for multiple steps of PC dendrite maturation in vivo.

Molecular identification of a new species of Rhigonema (Nematoda: Rhigonematidae) and phylogenetic relationships within the infraorder Rhigonematomorpha.

BACKGROUND: The infraorder Rhigonematomorpha comprises a group of obligate parasitic nematodes of millipedes (Arthropoda: Diplopoda). The current species identification of Rhigonematomorpha nematodes remains mainly based on morphological features, with molecular-based identification still in its infancy. Also, current knowledge of the phylogeny of Rhigonematomorpha is far from comprehensive. METHODS: The morphology of Rhigonematomorpha nematodes belonging to the genus Rhigonema, collected from the millipede Spirobolus bungii Brandt (Diplopoda: Spirobolida) in China, was studied in detail using light and scanning electron microscopy. Five different genetic markers, including the nuclear small ribosomal subunit (18S), internal transcribed spacer (ITS) and large ribosomal subunit (28S) regions and the mitochondrial cox1 and cox2 genes of these Rhigonematomorpha nematodes collected from China and Rhigonema naylae collected from Japan were sequenced and analyzed using Bayesian inference (BI) and Assemble Species by Automatic Partitioning (ASAP) methods. Phylogenetic analyses that included the most comprehensive taxa sampling of Rhigonematomorpha to date were also performed based on the 18S + 28S genes using maximum likelihood (ML) and BI methods. RESULTS: The specimens of Rhigonema collected from S. bungii in China were identified as a new species, Rhigonema sinense n. sp. Striking variability in tail morphology was observed among individuals of R. sinense n. sp. ASAP analyses based on the 28S, ITS, cox1 and cox2 sequences supported the species partition of R. sinense n. sp. and R. naylae, but showed no evidence that the different morphotypes of R. sinense n. sp. represent distinct genetic lineages. BI analyses also indicated that R. sinense n. sp. represents a separated species from R. naylae based on the cox1 and cox2 genes, but showed that R. naylae nested in samples of R. sinense n. sp. based on the ITS and 28S data. Phylogenetic results showed that the representatives of Rhigonematomorpha formed two large clades. The monophyly of the families Carnoyidae and Ichthyocephalidae and the genus Rhigonema was rejected. The representatives of the family Ransomnematidae clustered together with the family Hethidae with strong support. CONCLUSIONS: A new species of Rhigonematomorpha, R. sinense n. sp. is described based on morphological and molecular evidence. ASAP analyses using 28S, ITS, cox1 and cox2 data indicate the striking variability in tail morphology of R. sinense n. sp. as intraspecific variation, and also suggest that partial 28S, ITS, cox1 and cox2 markers are effective for molecular identification of Rhigonematomorpha nematodes. The phylogenetic results support the traditional classification of Rhigonematomorpha into the two superfamilies Rhigonematoidea and Ransomnematoidea, and indicate that the families Carnoyidae and Ichthyocephalidae and the genus Rhigonema are non-monophyletic. The present phylogeny strongly supports resurrection of the family Brumptaemiliidae, and also indicates that the family Ransomnematidae is sister to the family Hethidae.

Lung fibrogenic microenvironment in mouse reconstitutes human alveolar structure and lung tumor.

A mesenchymal cell activation is a hallmark event of pulmonary fibrosis. Alveolar type 2 (AT2) cells are progenitor cells that maintain alveolar homeostasis, and their damage is assumed to be an initiating event for pulmonary fibrosis. However, the interaction between the lung fibrogenic microenvironment and AT2 cell dynamics remains to be elucidated. Here, we report a unique role of the lung fibrogenic microenvironment, where cell type-specific tissue reconstruction is achieved by exogenous cell transplantation. We found that in the lung fibrogenic microenvironment the AT2 cell pool was depleted, whereas mesenchymal cells could promote intact AT2 cell proliferation in vitro. Furthermore, exogenously transplanted AT2 cells formed alveolar colonies and ameliorated pulmonary fibrosis. Exogenous tumor cells formed tumor nests with relevant histological and transcriptional properties. Human primary cells were adaptable to this microenvironment, facilitating epithelial cell-targeted therapy in pulmonary fibrosis and the establishment of patient-derived xenografts for precision medicine in lung cancer.

Visualizing interface-specific chemical bonds in adhesive bonding of carbon fiber structural composites using soft X-ray microscopy.

Adhesion is a technology for assembling carbon fiber (CF) reinforced polymer (CFRP), enabling them to maintain their lightweight and high-stiffness properties. Despite the importance of adhesion, the lack of a molecular-level understanding of the adhesion mechanisms has limited the reliability of adhesion for use in next-generation aircraft and automobiles. Here, we focused on the chemical-state distribution at a practical adhesive interface composed of an epoxy-based adhesive film bonded to an epoxy-based CF matrix. By fluorinating the OH group, we succeeded in visualizing the chemical state at the CF-matrix/adhesive interface using soft X-ray microscopy. The soft X-ray images exhibited a decrease in OH-related signals at the interface due to the local chemical interaction at the epoxy-epoxy adhesive interface. We also found that the N and O Kα signals were observable at the CF's surface, indicating the presence of nitrogen- and oxygen-containing functional groups. Based on these observations, we discuss the molecular-level adhesion mechanism at the CF-matrix/adhesive interface.

Possible stochastic sex determination in Bursaphelenchus nematodes.

Sex determination mechanisms evolve surprisingly rapidly, yet little is known in the large nematode phylum other than for Caenorhabditis elegans, which relies on chromosomal XX-XO sex determination and a dosage compensation mechanism. Here we analyze by sex-specific genome sequencing and genetic analysis sex determination in two fungal feeding/plant-parasitic Bursaphelenchus nematodes and find that their sex differentiation is more likely triggered by random, epigenetic regulation than by more well-known mechanisms of chromosomal or environmental sex determination. There is no detectable difference in male and female chromosomes, nor any linkage to sexual phenotype. Moreover, the protein sets of these nematodes lack genes involved in X chromosome dosage counting or compensation. By contrast, our genetic screen for sex differentiation mutants identifies a Bursaphelenchus ortholog of tra-1, the major output of the C. elegans sex determination cascade. Nematode sex determination pathways might have evolved by "bottom-up" accretion from the most downstream regulator, tra-1.

Molecular mechanisms regulating the spatial configuration of neurites.

Precise neural networks, composed of axons and dendrites, are the structural basis for information processing in the brain. Therefore, the correct formation of neurites is critical for accurate neural function. In particular, the three-dimensional structures of dendrites vary greatly among neuron types, and the unique shape of each dendrite is tightly linked to specific synaptic connections with innervating axons and is correlated with its information processing. Although many systems are involved in neurite formation, the developmental mechanisms that control the orientation, size, and arborization pattern of neurites definitively defines their three-dimensional structure in tissues. In this review, we summarize these regulatory mechanisms that establish proper spatial configurations of neurites, especially dendrites, in invertebrates and vertebrates.

Morphological and molecular characterization of two species of nematodes (Oxyuridomorpha: Thelastomatoidea: Protrelloididae, Thelastomatidae) parasitic in the cockroach Blaberus discoidalis Serville (Blattaria: Blaberidae) from Cuba.

Protrelleta floridana Chitwood, 1932 and Cranifera cranifera (Chitwood, 1932) (Oxyuridomorpha: Thelastomatoidea: Protrelloididae, Thelastomatidae) are recorded for the first time in Cuba. These nematodes were found to parasitize the cockroach Blaberus discoidalis Serville (Blattaria: Blaberidae), constituting a new host record for them. Both species are redescribed with the aid of scanning electron microscopy and the arrangement of the copulatory papillae of the males of P. floridana is amended. The present specimens coincide in their morphology and most of the measurements with the type populations from USA and the records from Costa Rica, with only minor differences. The molecular phylogeny was inferred by mean of the D2-D3 domain of the 28S rDNA and the Cuban P. floridana and C. cranifera form monophyletic clades with sequences of both taxa from Costa Rica as well as a sequence of C. cranifera from Russia. In the case of C. cranifera its phylogeny and that of its blaberid hosts reveal coevolutionary relationships.

Simulation Study of the Effects of Interfacial Bonds on Adhesion and Fracture Behavior of Epoxy Resin Layers.

The adhesion and fracture behavior of tetraglycidyl-4,4'-diaminodiphenylmethane (TGDDM)/4,4'-diaminodiphenyl sulfone (44DDS)-bisphenol A diglycidyl ether (DGEBA)/44DDS layer interfaces were investigated by molecular dynamics (MD) simulation, mainly focusing on the role of covalent and noncovalent interactions. To accurately investigate the bond dissociation processes, the force field parameters of several bond potentials of the epoxy resin polymers were optimized by density functional theory calculations. In the MD simulations under a tensile load, small voids gradually developed without covalent bond dissociation in the plateau region. In the final large strain region, the stress rapidly increased with bond breaking, leading to failure. When the chemical bonds across the interface between the two layers were removed, the stress-strain curve in the initial elastic region was almost the same as that with interfacial bonds. This showed that the nonbonded interactions governed adhesion strength in the initial elastic region. In contrast, the bonded interactions at interfaces played important roles in the hardening regions because the bonded interactions made the major contribution to the fracture energies. We also investigated the effect of the etherification reaction in cross-linking. It was found that the etherification reaction mainly contributed to the behavior in the late region with large strain. These simulation results revealed that the nonbonded interactions, especially, van der Waals interactions, are important factors for adhesion of the different polymer layers in the small strain region up to the yield point.

Morphological and molecular characterization of Buzionema lutgardae n. sp. (Nematoda: Oxyuridomorpha: Thelastomatidae) from the cockroach Byrsotria sp. (Blattaria: Blaberidae) in Cuba.

Buzionema lutgardae n. sp. (Nematoda: Oxyuridomorpha: Thelastomatidae) is described from the cockroach Byrsotria sp. (Blattaria: Blaberidae), endemic to Cuba. Females of B. lutgardae n. sp. are shorter than those of B. validum Kloss, 1966 (16002150 µm vs. 31313378 µm), but the oesophagus is comparatively longer (b = 2.963.77 vs. 4.654.87). The lateral alae of the new species extend from ca. the midpoint of the cylindrical part of the procorpus to the level of the anus in contrast to the base of the basal bulb to the level of the anus in B. validum. The males of B. lutgardae n. sp. are shorter than those of B. validum (780940 µm vs. 11771423 µm) and their lateral alae end at some distance before the cloaca instead the level of the cloaca in B. validum. The phylogeny of B. lutgardae n. sp. is inferred by the D2-D3 domains of the 28S rDNA. B. lutgardae n. sp. and B. validum form a monophyletic clade with strong nodal support, as sister-group of the genus Leidynema Schwenck in Travassos, 1929.

Symbiosis of the millipede parasitic nematodes Rhigonematoidea and Thelastomatoidea with evolutionary different origins.

BACKGROUND: How various host-parasite combinations have been established is an important question in evolutionary biology. We have previously described two nematode species, Rhigonema naylae and Travassosinema claudiae, which are parasites of the xystodesmid millipede Parafontaria laminata in Aichi Prefecture, Japan. Rhigonema naylae belongs to the superfamily Rhigonematoidea, which exclusively consists of parasites of millipedes. T. claudiae belongs to the superfamily Thelastomatoidea, which includes a wide variety of species that parasitize many invertebrates. These nematodes were isolated together with a high prevalence; however, the phylogenetic, evolutionary, and ecological relationships between these two parasitic nematodes and between hosts and parasites are not well known. RESULTS: We collected nine species (11 isolates) of xystodesmid millipedes from seven locations in Japan, and found that all species were co-infected with the parasitic nematodes Rhigonematoidea spp. and Thelastomatoidea spp. We found that the infection prevalence and population densities of Rhigonematoidea spp. were higher than those of Thelastomatoidea spp. However, the population densities of Rhigonematoidea spp. were not negatively affected by co-infection with Thelastomatoidea spp., suggesting that these parasites are not competitive. We also found a positive correlation between the prevalence of parasitic nematodes and host body size. In Rhigonematoidea spp., combinations of parasitic nematode groups and host genera seem to be fixed, suggesting the evolution of a more specialized interaction between Rhigonematoidea spp. and their host. On the other hand, host preference of Thelastomatoidea spp. was not specific to any millipede species, indicating a non-intimate interaction between these parasites and their hosts. CONCLUSIONS: The two nematode superfamilies, Rhigonematoidea and Thelastomatoidea, have phylogenetically distinct origins, and might have acquired xystodesmid millipede parasitism independently. Currently, the two nematodes co-parasitize millipedes without any clear negative impact on each other or the host millipedes. Our study provides an example of balanced complex symbioses among parasitic nematodes and between parasitic nematodes and host millipedes, which have been established over a long evolutionary history.

Vascularization of human brain organoids.

Human brain organoids are three-dimensional tissues that are generated in vitro from pluripotent stem cells and recapitulate the early development of the human brain. Brain organoids consist mainly of neural lineage cells, such as neural stem/precursor cells, neurons, astrocytes, and oligodendrocytes. However, all human brain organoids lack vasculature, which plays indispensable roles not only in brain homeostasis but also in brain development. In addition to the delivery of oxygen and nutrition, accumulating evidence suggests that the vascular system of the brain regulates neural differentiation, migration, and circuit formation during development. Therefore, vascularization of human brain organoids is of great importance. Current trials to vascularize various organoids include the adjustment of cultivation protocols, the introduction of microfluidic devices, and the transplantation of organoids into immunodeficient mice. In this review, we summarize the efforts to accomplish vascularization and perfusion of brain organoids, and we discuss these attempts from a forward-looking perspective.

Morphological and molecular characterization of Klossnema viguerasi n. sp. (Nematoda: Oxyuridomorpha: Hystrignathidae) from a Cuban passalid beetle (Coleoptera: Passalidae), first record of the genus for Cuba.

Klossnema viguerasi n. sp. (Nematoda: Oxyuridomorpha: Hystrignathidae) is described from the passalid beetle Antillanax pertyi (Kaup, 1869), endemic to Cuba. The females of K. viguerasi n. sp. are morphologically similar but slightly longer than K. repentina Cordeiro Artigas, 1983 (1.143 mm vs. 1.000 mm). Both species differ in that K. viguerasi n. sp. has a longer procorpus (139 µm vs. 110 µm), isthmus (39 µm vs. 24 µm), and tail length (28 µm vs. 21 µm). The distance from the vulva to the anterior end is also longer in the new species (0.748 mm vs. 0.650 mm). The males of K. viguerasi n. sp. are larger than K. repentina (0.980 mm vs. 0.800 mm), but their isthmus is shorter (38 µm vs. 48 µm). New features of the cephalic end of both sexes, and copulatory papillae pattern of the males were observed by SEM and the generic diagnosis is emended in order to include such features. The phylogeny of K. viguerasi n. sp. is inferred by the analysis of the D2-D3 domains of the 28S rDNA and the 18S rDNA. This constitutes the first record of the genus Klossnema for the Cuban archipelago and the West Indies.

Exposure to particulate matter upregulates ACE2 and TMPRSS2 expression in the murine lung.

Coronavirus disease (COVID-19) is currently a serious global issue. Epidemiological studies have identified air pollutants, including particulate matter (PM), as a risk factor for COVID-19 infection and severity of illness, in addition to numerous factors such as pre-existing conditions, aging and smoking. However, the mechanisms by which air pollution is involved in the manifestation and/or progression of COVID-19 is still unknown. In this study, we used a mouse model exposed to crude PM, collected by the cyclone method, to evaluate the pulmonary expression of angiotensin-converting enzyme 2 (ACE2) and transmembrane protease serine type 2 (TMPRSS2), the two molecules required for the entry of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) into host cells. Multiplex immunohistochemical analysis revealed that exposure to PM increased the expression of these two molecules at the same site. Furthermore, image cytometry analysis revealed increased expression of these proteins, particularly, in the alveolar type 2 cells and macrophages, which are potential targets for SARS-CoV-2. Our findings provide an experimental evidence that exposure to PM may adversely affect the manifestation and progression of COVID-19, mediated by the impact of SARS-CoV-2 on the site of entry. The study results suggest that examining these effects might help to advance our understanding of COVID-19 and aid the development of appropriate social interventions.

Three new species of Longior Travassos Kloss, 1958 (Nematoda: Thelastomatoidea: Hystrignathidae) parasites of passalid beetles (Coleoptera: Passalidae) from Dominican Republic, Mexico and Colombia.

Three new species of the genus Longior Travassos Kloss, 1958 are described and illustrated, namely L. surieli n. sp. in Antillanax dominicanus (Doesburg, 1953) from the Dominican Republic, L. lamothei n. sp. in Passalus punctiger Le Peletier Serville, 1825 from Mexico and Colombia and L. zumpimito n. sp. in P. punctatostriatus Percheron, 1835 from Mexico. These constitute the first records of the genus Longior for the aforementioned countries, rising to nine species in the genus. The new species can be differentiated mainly by the length of their body, oesophagus and tail in both sexes, the extension of the lateral alae in the females and the morphology of the cephalic and posterior end in the males. The molecular phylogeny of the new taxa is inferred by the 28S and 18S rDNA and they form a monophyletic clade with other Longior species. The phylogeny of Longior and that of their passalid hosts reveal coevolutionary relationships. These patterns suggest that the phylogeny of Longior species is probably strongly influenced by the evolutionary trajectories of their passalid hosts.