Polyandry may mitigate the negative impact of reproductive interference among bumblebees in Japan.

In social hymenopterans, monandry of the queen is an ancestral trait, and polyandry is a derived trait. Polyandry of the queen is the norm in a limited number of lineages, such as honeybees, leaf-cutting ants, Pogonomyrmex ants, and Vespula wasps, which presumably provide fitness advantages for the whole colony. The queen of the introduced bumblebee, Bombus terrestris, is polyandrous in Japan, whereas it is monandrous in native regions. We hypothesize that polyandry can evolve in a process that avoids the negative impacts of reproductive interference caused by interspecific mating and conducted genetic studies of the invasive species B. terrestris and two native subspecies, Bombus hypocrita sapporoensis and Bombus hypocrita hypocrita, in Japan. Our results revealed that although the native queens of B. hypocrita hypocrita allopatric with B. terrestris were strictly monandrous, the native queens of B. hypocrita sapporoensis sympatric with B. terrestris were polyandrous. These results suggested that the queens of native B. hypocrita sapporoensis do not experience negative impacts on interspecific mating from the invasive B. terrestris. We discuss the possibility that reproductive interference is a driving force in selection for multiple mating through an arms race between sympatric species.

Reproductive interference by alien hornet Vespa velutina threatens the native populations of Vespa simillima in Japan.

The yellow-legged hornet Vespa velutina has become one of the major alien species in European and East Asian countries. As in its homeland, the invading V. velutina is reported as the major predator of honeybees and is becoming a threat to beekeeping in Europe. However, it remains unknown how V. velutina might affect native hornets when it invades Asia, where a large number of Vespa species are distributed. Thus, by analyzing the mitochondrial cytochrome c oxidase subunit I gene, we investigated whether interspecific mating occurs between V. velutina and Japanese native Vespa spp. Our results showed that the alien V. velutina causes reproductive interference in Japanese native hornet Vespa simillima. Forty-three percent of native V. simillima queens had the sperm of V. velutina males, and among the all V. simillima queens analyzed, 28% only had V. velutina sperm. We did not find evidence of V. velutina queens having the sperm of V. simillima males. These findings suggest that reproductive interference by V. velutina males poses a threat to the native V. simillima populations. A decline of V. simillima may also negatively affect other insects that interact with V. simillima.

Impact of Statins Therapy for Ischemic Heart Disease Patients with Low-Density Lipoprotein Cholesterol Levels Less Than 100 mg/dL.

BACKGROUND: The objective of this study was to determine whether the use of statins prevents the progression of ischemic heart disease (IHD) in patients with low levels of low-density lipoprotein cholesterol (LDL-C). METHODS: We reviewed data obtained from IHD patients who underwent first percutaneous coronary intervention (PCI). Patients underwent follow-up coronary angiography (re-CAG) after PCI. However, only patients with LDL-C levels less than 100 mg/dL at PCI were included in this study. Ultimately, 92 patients were enrolled. All patients were divided into two groups: 1) patients who were treated with statins (n = 69), and 2) patients who were not treated with statins (n = 23). RESULTS: The two groups had similar LDL-C levels at PCI. At re-CAG, the ratio of patients who underwent PCI for de novo lesion in the statin group was lower than that in the non-statin group (12% vs. 48%) (p < 0.001). In multiple regression analysis, statin usage and LDL-C level at PCI were independent predictors of the ratio of patients undergoing PCI for de novo lesion. CONCLUSIONS: Statins therapy for patients whose LDL-C levels are less than 100 mg/dL has a beneficial effect on secondary prevention of IHD.

Herbivore community promotes trait evolution in a leaf beetle via induced plant response.

Several recent studies have emphasised that community composition alters species trait evolution. Here, we demonstrate that differences in composition of local herbivore communities lead to divergent trait evolution of the leaf beetle Plagiodera versicolora through plant-mediated indirect interactions. Our field surveys, genetic analyses and community-manipulation experiments show that herbivore community composition determines the degree of herbivore-induced regrowth of willows (Salicaceae), which in turn, promotes the divergent evolution of feeding preference in the leaf beetle from exclusive preference for new leaves to a lack of preference among leaf-age types. Regrowth intensity depends both on the differential response of willows to different herbivore species and the integration of those herbivore species in the community. Because herbivore-induced regrowth involves phenological changes in new leaf production, leaf beetle populations develop divergent feeding preferences according to local regrowth intensity. Therefore, herbivore community composition shapes the selection regime for leaf beetle evolution through trait-mediated indirect interactions.

Cross-mating between the alien bumblebee Bombus terrestris and two native Japanese bumblebees, B. hypocrita sapporensis and B. cryptarum florilegus, in the Nemuro Peninsula, Japan.

The rapid naturalization of Bombus terrestris across the Nemuro Peninsula has led to a decline in two closely related native Japanese species, namely Bombus hypocrita sapporensis and Bombus cryptarum florilegus, both belonging to the common subgenus Bombus. Although it is widely believed that cross-mating of native and non-native species is influenced by the common male sex pheromone in this region, no study has been conducted to substantiate this claim. Thus, we investigated the cross-activities of male sex pheromones between native and non-native bumblebees, as well as the frequencies of cross-mating, using chemical and DNA assays. Our gas chromatography-electroantennographic detector analyses and behavioral tests revealed the presence of sex pheromonal cross-activities between B. terrestris and the two Japanese bumblebees species. Furthermore, DNA analyses revealed the occurrence of cross-mating between native and non-native species in the Nemuro Peninsula. Overall, these results indicate the immediate need for conservation measures to safeguard Japanese bumblebee populations in the Nemuro Peninsula.

Genetic differentiation and local adaptation of the Japanese honeybee, Apis cerana japonica.

We examine the population genetic structure and divergence among the regional populations of the Japanese honeybee, Apis cerana japonica, by re-sequencing the genomes of 105 individuals from the three main Japanese islands with diverse climates. The genetic structure results indicated that these individuals are distinct from the mainland Chinese A. cerana samples. Furthermore, population structure analyses have identified three genetically distinct geographic regions in Japan: Northern (Tohoku-Kanto-Chubu districts), Central (Chugoku district), and Southern (Kyushu district). In some districts, "possible non-native" individuals, likely introduced from other regions in recent years, were discovered. Then, genome-wide scans were conducted to detect candidate genes for adaptation by two different approaches. We performed a population branch statistics (PBS) analysis to identify candidate genes for population-specific divergence. A latent factor mixed model (LFMM) was used to identify genes associated with climatic variables along a geographic gradient. The PBSmax analysis identified 25 candidate genes for population-specific divergence whereas the LFMM analysis identified 73 candidate genes for adaptation to climatic variables along a geographic gradient. However, no common genes were identified by both methods.

Formation of Amino Acids and Carboxylic Acids in Weakly Reducing Planetary Atmospheres by Solar Energetic Particles from the Young Sun.

Life most likely started during the Hadean Eon; however, the environmental conditions which contributed to the complexity of its chemistry are poorly known. A better understanding of various environmental conditions, including global (heliospheric) and local (atmospheric, surface, and oceanic), along with the internal dynamic conditions of the early Earth, are required to understand the onset of abiogenesis. Herein, we examine the contributions of galactic cosmic rays (GCRs) and solar energetic particles (SEPs) associated with superflares from the young Sun to the formation of amino acids and carboxylic acids in weakly reduced gas mixtures representing the early Earth's atmosphere. We also compare the products with those introduced by lightning events and solar ultraviolet light (UV). In a series of laboratory experiments, we detected and characterized the formation of amino acids and carboxylic acids via proton irradiation of a mixture of carbon dioxide, methane, nitrogen, and water in various mixing ratios. These experiments show the detection of amino acids after acid hydrolysis when 0.5% (v/v) of initial methane was introduced to the gas mixture. In the set of experiments with spark discharges (simulation of lightning flashes) performed for the same gas mixture, we found that at least 15% methane was required to detect the formation of amino acids, and no amino acids were detected in experiments via UV irradiation, even when 50% methane was used. Carboxylic acids were formed in non-reducing gas mixtures (0% methane) by proton irradiation and spark discharges. Hence, we suggest that GCRs and SEP events from the young Sun represent the most effective energy sources for the prebiotic formation of biologically important organic compounds from weakly reducing atmospheres. Since the energy flux of space weather, which generated frequent SEPs from the young Sun in the first 600 million years after the birth of the solar system, was expected to be much greater than that of GCRs, we conclude that SEP-driven energetic protons are the most promising energy sources for the prebiotic production of bioorganic compounds in the atmosphere of the Hadean Earth.

Floating chip mounting system driven by repulsive force of permanent magnets for multiple on-site SPR immunoassay measurements.

We have developed a measurement chip installation/removal mechanism for a surface plasmon resonance (SPR) immunoassay analysis instrument designed for frequent testing, which requires a rapid and easy technique for changing chips. The key components of the mechanism are refractive index matching gel coated on the rear of the SPR chip and a float that presses the chip down. The refractive index matching gel made it possible to optically couple the chip and the prism of the SPR instrument easily via elastic deformation with no air bubbles. The float has an autonomous attitude control function that keeps the chip parallel in relation to the SPR instrument by employing the repulsive force of permanent magnets between the float and a float guide located in the SPR instrument. This function is realized by balancing the upward elastic force of the gel and the downward force of the float, which experiences a leveling force from the float guide. This system makes it possible to start an SPR measurement immediately after chip installation and to remove the chip immediately after the measurement with a simple and easy method that does not require any fine adjustment. Our sensor chip, which we installed using this mounting system, successfully performed an immunoassay measurement on a model antigen (spiked human-IgG) in a model real sample (non-homogenized milk) that included many kinds of interfering foreign substances without any sample pre-treatment. The ease of the chip installation/removal operation and simple measurement procedure are suitable for frequent on-site agricultural, environmental and medical testing.

Passive fluidic chip composed of integrated vertical capillary tubes developed for on-site SPR immunoassay analysis targeting real samples.

We have successfully developed a surface plasmon resonance (SPR) measurement system for the on-site immunoassay of real samples. The system is composed of a portable SPR instrument (290 mm(W) × 160 mm(D) × 120 mm(H)) and a microfluidic immunoassay chip (16 mm(W) × 16 mm(D) × 4 mm(H)) that needs no external pump system. An integrated vertical capillary tube functions as a large volume (150 µL) passive pump and a waste reservoir that has sufficient capacity for several refill operations. An immunoassay was carried out that employed the direct injection of a buffer and a test sample in sequence into a microfluidic chip that included 9 antibody bands and 10 reference reagent bands immobilized in the flow channel. By subtracting a reliable averaged reference sensorgram from the antibody, we effectively reduced the influence of the non-specific binding, and then our chip successfully detected the specific binding of spiked IgG in non-homogeneous milk. IgG is a model antigen that is certain not to be present in non-homogeneous milk, and non-homogeneous milk is a model of real sample that includes many interfering foreign substances that induce non-specific binding. The direct injection of a real sample with no pretreatment enabled us to complete the entire immunoassay in several minutes. This ease of operation and short measuring time are acceptable for on-site agricultural, environmental and medical testing.

Photostability of Iiovaline and its precursor 5-Ethyl-5- methylhydantoin exposed to simulated space radiations.

Aqueous solutions of isovaline and its precursor molecule, 5-ethyl-5-methylhydantoin, were irradiated with ultraviolet and γ-ray photons, to evaluate their structural stability against space radiation. The degree of photolysis was measured and irradiation products were identified using chiral, reversed-phase and ion-exchange high-performance liquid chromatography. The experimental results show that the degree of photolysis of 5-ethyl-5-methylhydantoin is more significant than that of isovaline under ultraviolet light irradiation, while the results under γ-ray irradiation are the opposite. As the products of isovaline photolysis, aspartic acid, serine, glutamic acid and alanine were dominantly detected.

Complete mitochondrial DNA sequence of the Eastern Asian catfish Silurus asotus (Siluriformes: Siluridae) from Lake Biwa in Japan.

The east Asian catfish Silurus asotus is a common species living in fresh water in Japan. The complete mitochondrial genome of the S. asotus from Lake Biwa in Japan was analyzed using next-generation sequencing. The mitochondrial genome of S. asotus was identified as a 16,515 bp circular molecule containing 13 protein-coding genes (PCGs), 22 tRNA genes, and two rRNA genes, along with one A + T-rich control region. The AT content was 56.1%. Start codons ATG and GTG were found in 13 PCGs. Stop codons TAA, TAG, and AGA were observed in 13 PCGs. The heavy (H)-strand was predicted to have 12 PCGs and 14 tRNA and two rRNA genes, while the light (L)-strand was predicted to contain one PCGs and eight tRNA genes. The molecular phylogenetic analysis showed that S. asotus from Lake Biwa is genetically similar to S. asotus from China.

Effectiveness of dental checkups incorporating tooth brushing instruction.

The purpose of this study was to compare the effectiveness of dental checkups incorporating tooth-brushing instruction (TBI) with that of conventional dental checkups. A team consisting of one dentist and three dental hygienists saw an average of 60 employees per day on-site at an airline company. The patient's teeth were stained with a disclosing tablet and the results recorded on a Plaque Control Record (PCR) chart. The patient was then given TBI. After recording the relevant data, including TBI given and PCR scores, the charts were stored. Checkups were performed in a total of 3,854 patients between 2001 and 2005 and changes in annual scores investigated. In addition, annual shifts in mean score in patients receiving checkups over all five years were compared with those in patients receiving checkups for the first time in each of the five years. The mean score in patients receiving a checkup in 2001 was 35.1%, declining by 2.6 points to 32.5% in 2005. Among patients receiving checkups over all five years, the mean score in 2001 was 34.0%, declining by 11.2 points to 22.8% in 2005. Over the five-year period, the mean score in patients receiving checkups was 34.1%. In patients receiving checkups over all five years, the proportion with PCR scores <30% increased each year. This was because the number of patients with PCR scores ≥60% decreased each year. These findings suggest that TBI is effective in reducing poor plaque control. When compared with in patients who had not received TBI, five consecutive years of checkups was clearly effective. These results indicate that checkups incorporating TBI are more effective than conventional dental checkups that simply check for caries. In future, this type of checkup should contribute to improved preventative dentistry with minimal intervention.

Complete mitochondrial DNA sequence of the Japanese endemic catfish Silurus biwaensis (Siluriformes: Siluridae) from Lake Biwa.

The Japanese endemic catfish Silurus biwaensis is distributed only in Lake Biwa and Yodo river drainages. There are four species of the genus Silurus in Japan, of which S. biwaensis has a most limited distribution. This catfish needs to be collected for DNA data owing to the lack of information related to its phylogenetic relationship. Here, the complete mitochondrial genome of the S. biwaensis from Lake Biwa in Japan was analyzed using next-generation sequencing. The mitochondrial genome of S. biwaensis was identified as a 16,531 bp circular molecule containing 13 protein-coding genes (PCGs), 22 tRNA genes, and 2 rRNA genes, along with one A + T-rich control region. The AT content was 55.83%. The heavy (H)-strand was predicted to have 12 PCGs, 14 tRNA, and 2 rRNA genes, whereas the light (L)-strand was predicted to contain one PCG and eight tRNA genes. The start codons ATG, ATC, and GTG were found in 13 PCGs. The stop codons TAA, TAG, and AGA were observed in all PCGs, except CytB and COX3. All tRNA genes formed typical cloverleaf secondary structures. The molecular phylogenetic relationships estimated using 13 PCGs (maximum-likelihood method) indicated that S. biwaensis is genetically distinct from the sympatric species S. asotus and S. lithophilus. This result clearly indicated that S. biwaensis is a valid species.

Complete mitochondrial DNA sequence of the Japanese endemic catfish Silurus lithophilus (Siluriformes: Siluridae).

The Japanese endemic catfish Silurus lithophilus is distributed only in Lake Biwa, Lake Yogo, and their river drainages. There are four species of the genus Silurus in Japan, of which S. lithophilus has a limited distribution. This catfish needs to be collected for DNA data owing to the lack of information related to its conservation. Here, the complete mitochondrial genome of the S. lithophilus from Lake Biwa in Japan was analyzed using next-generation sequencing. The mitochondrial genome of S. lithophilus was identified as a 16,524 bp circular molecule containing 13 protein-coding genes (PCGs), 22 tRNA genes, and 2 rRNA genes, along with one A + T-rich control region. The AT content was 55.89%. The heavy (H)-strand was predicted to have 12 PCGs and 15 tRNA, and 2 rRNA genes, whereas the light (L)-strand was predicted to contain one PCG and seven tRNA genes. The start codons ATG and GTG were found in 13 PCGs. The stop codons TAA, TAG, and AGA were observed in all PCGs, except CytB. All tRNA genes formed typical cloverleaf secondary structures. The molecular phylogenetic relationship, inferred using 13 PCGs (based on the maximum likelihood), was consistent with that reported in previous studies, which predicted a sister relationship between S. lithophilus and S. asotus. The results also clearly indicated that S. lithophilus is a valid species.

Complete mitochondrial DNA sequence of the giant water bug Kirkaldyia deyrolli (Hemiptera: Belostomatidae).

Giant water bugs (genus Kirkaldyia and Lethocerus) are well known species from aquatic habitats throughout the world's subtropical and tropical areas. Only one species of the Kirkaldyia deyrolli is distributed in Japan. K. deyrolli has been designated as second category rare species according Law for Conservation of Endangered Species of Wild Fauna and Flora in Japan, in 2019. We analyzed, for the first time, the complete mitochondrial genomes of the giant water bug K. deyrolli from Japan using next-generation sequencing. The mitochondrial genome was a circular and 15,579 bp molecule that included 13 protein-coding genes (PCGs), 22 tRNA genes, and two ribosomal RNA genes, along with one AT-rich control region. The AT content value was 69.55%. The heavy strand was predicted to have nine PCGs and 15 tRNA genes, whereas the light strand was predicted to contain four PCGs, seven tRNA genes, and two rRNA genes. Start codons were variable for all PCGs: four ATA, three ATC, four ATG, two ATT genes as the start codon. Stop codons were of two types: TAA for 12 genes and TAG for one gene. Incomplete stop codon T was identified. The molecular phylogenetic relationship, inferred using 13 PCGs, was consistent with that reported in previous studies, which predicted a sister relationship to the genus Lethocerus.

Complete mitochondrial DNA sequence of the parasitic honey bee mite Varroa destructor (Mesostigmata: Varroidae).

Varroa destructor is a parasite mite of the eastern honey bee Apis cerana, which is native to Asia. The European honey bee Apis mellifera was imported to Asia from Europe and the USA for apiculture in the 19th century. In a short period of time, V. destructor parasitized the artificially introduced honey bees. Varroa destructor was estimated to have spread around the world with A. mellifera when it was exported from Asia to locations worldwide about 50 years ago. The mitochondrial DNA of the parasitic honey bee mite V. destructor was analyzed using next-generation sequencing. The complete mitochondrial genome of V. destructor was identified as a 16,476-bp circular molecule containing 13 protein-coding genes (PCGs), 22 tRNA genes, two rRNA genes, and one AT-rich control region. The heavy strand was predicted to have nine PCGs and 13 tRNA genes, whereas the light strand was predicted to contain four PCGs, nine tRNA genes, and two rRNA genes. All PCGs began with ATA as the start codon, except COIII and CytB, which had ATG as the start codon. Stop codons were of two types: TAA for eight genes and TAG for five genes. Molecular phylogenetic analysis revealed that V. destructor from Japan was genetically distant from that of France. A high base substitution rate of 2.82% was also confirmed between the complete mitochondrial DNA sequences of V. destructor from Japan and the USA, suggesting that one Varroa mite strain found in the USA is not from Japan.

Complete mitochondrial DNA sequence of the East Asian minnow, Pungtungia herzi (Actinopterygii: Cypriniformes).

Some populations of the East Asian minnow Pungtungia herzi Herzenstein, 1892, which are naturally distributed in western Japan, have declined and are considered endangered. However, P. herzi has become a domestic invasive species in eastern Japan. Thus, knowledge of genetic features and phylogenetic relationships of P. herzi is important for conservation of this species and understanding its impact on ecosystems. We analyzed the complete mitochondrial genome using next generation sequencing of the East Asian minnow P. herzi from Yodo River, Osaka Prefecture, Japan. The mitochondrial genome of P. herzi consists of a circular molecule of 16,599 bp that includes 13 protein-coding genes (PCGs), 22 tRNA genes, two rRNA genes, and one control region. The heavy (H)-strand was predicted to have 12 PCGs, 14 tRNA, and two rRNA genes, while the light (L)-strand was predicted to contain one PCGs and eight tRNA genes. The average AT content was 57.68%. The genes ATP8 and ATP6, ATP6 and COIII, ND4L and ND4, and ND5 and ND6 shared seven, one, seven, and four nucleotides, respectively. The initiation codons ATG and GTG were found in 12 and one genes, respectively. The termination codons TAA, TAG, incomplete TA-, and single T-- were observed in nine, one, one, and two genes, respectively. All the tRNA genes possessed a cloverleaf secondary structure. The phylogenetic relationships inferred using 13 PCGs (based on the maximum likelihood) were consistent with previous studies that predicted interrelationships of Cypriniformes.

A revision of subspecies structure of western honey bee Apis mellifera.

The taxonomy of honey bee A. mellifera contains a lot of issues due to the specificity of population structure, features of biology and resolutions of honey bee subspecies discrimination methods. There are a lot of transition zones between ranges of subspecies which led to the gradual changes of characteristics among neighbor subspecies. The modern taxonomic pattern of honey bee Apis mellifera is given in this paper. Thirty-three distinct honey bee subspecies are distributed across all Africa (11 subspecies), Western Asia and the Middle East (9 subspecies), and Europe (13 subspecies). All honey bee subspecies are subdivided into 5 evolutionary lineages: lineage A (10 subspecies) and its sublineage Z (3 subspecies), lineage M (3 subspecies), lineage C (10 subspecies), lineage O (3 subspecies), lineage Y (1 subspecies), lineage C or O (3 subspecies).

Complete mitochondrial DNA sequence of the paper wasp Polistes riparius (Hymenoptera: Vespidae).

The paper wasp Polistes riparius is distributed in cold regions of northern East Asia to Russia. P. riparius are characterized by having longer cells than those of the closely related P. chinensis, which has a similar life history, as an adaptation to cold regions. The phylogenetic relationships of paper wasps have recently been studied; however, the genetic diversity and population structure of P. riparius has not been determined. The present study is the first to analyze the complete mitochondrial genome using next generation sequencing of P. riparius collected from Sapporo, Hokkaido Prefecture, Japan. The genome consisted of a closed loop that was 16,383 bp-long and included 13 protein coding genes (PCGs), 22 tRNA genes, two rRNA genes, and one AT-rich control region. The average AT content was 84.54%. The heavy (H)-strand was predicted to have 12 PCGs and 14 tRNA genes, while the light (L)-strand was predicted to contain one PCGs, eight tRNA genes, and two rRNA genes. All PCGs started with ATG. Stop codons were of two types: TAA for 11 genes (ND1, ND2, ND3, ND4L, ND5, ND6, COXI, COXII, COXIII, COB, ATP6 and ATP8) and TAG for two genes (ND3 and ND4). The molecular phylogenetic relationship based on the maximum likelihood method using 13 PCGs was consistent with some previous studies in which a closely relationship between P. riparius and P. jokahamae.

Chirality emergence in thin solid films of amino acids by polarized light from synchrotron radiation and free electron laser.

One of the most attractive hypothesis for the origin of homochirality in terrestrial bioorganic compounds is that a kind of "chiral impulse" as an asymmetric excitation source induced asymmetric reactions on the surfaces of such materials such as meteorites or interstellar dusts prior to the existence of terrestrial life (Cosmic Scenario). To experimentally introduce chiral structure into racemic films of amino acids (alanine, phenylalanine, isovaline, etc.), we irradiated them with linearly polarized light (LPL) from synchrotron radiation and circularly polarized light (CPL) from a free electron laser. After the irradiation, we evaluated optical anisotropy by measuring the circular dichroism (CD) spectra and verified that new Cotton peaks appeared at almost the same peak position as those of the corresponding non-racemic amino acid films. With LPL irradiation, two-dimensional anisotropic structure expressed as linear dichroism and/or linear birefringence was introduced into the racemic films. With CPL irradiation, the signs of the Cotton peaks exhibit symmetrical structure corresponding to the direction of CPL rotation. This indicates that some kinds of chiral structure were introduced into the racemic film. The CD spectra after CPL irradiation suggest the chiral structure should be derived from not only preferential photolysis but also from photolysis-induced molecular structural change. These results suggest that circularly polarized light sources in space could be associated with the origin of terrestrial homochirality; that is, they would be effective asymmetric exciting sources introducing chiral structures into bio-organic molecules or complex organic compounds.