Horizontal gene transfer from diverse bacteria to an insect genome enables a tripartite nested mealybug symbiosis.

The smallest reported bacterial genome belongs to Tremblaya princeps, a symbiont of Planococcus citri mealybugs (PCIT). Tremblaya PCIT not only has a 139 kb genome, but possesses its own bacterial endosymbiont, Moranella endobia. Genome and transcriptome sequencing, including genome sequencing from a Tremblaya lineage lacking intracellular bacteria, reveals that the extreme genomic degeneracy of Tremblaya PCIT likely resulted from acquiring Moranella as an endosymbiont. In addition, at least 22 expressed horizontally transferred genes from multiple diverse bacteria to the mealybug genome likely complement missing symbiont genes. However, none of these horizontally transferred genes are from Tremblaya, showing that genome reduction in this symbiont has not been enabled by gene transfer to the host nucleus. Our results thus indicate that the functioning of this three-way symbiosis is dependent on genes from at least six lineages of organisms and reveal a path to intimate endosymbiosis distinct from that followed by organelles.

Nursing researchers' concern about research activities during the COVID-19 pandemic: A secondary analysis of longitudinal survey data in Japan.

The coronavirus disease (COVID-19) pandemic has negatively affected research activities across various fields. This study aimed to determine nursing researchers' concerns about research activities during the COVID-19 pandemic in Japan and subsequent changes brought on by it. For this study, we conducted descriptive statistics and text mining analyses using data from two surveys conducted by the Japan Academy of Nursing Science (JANS) in the early days of the pandemic (first survey: mid-2020) and after 2 years (second survey: early 2022). Concerns about research activities were observed in 89% and 80% of the nursing researchers in the first and second surveys, respectively. Furthermore, concerns about "Difficulty in collecting research data" and "Content and quality of your research" were stronger in the second survey. Text mining analyses revealed that in the first survey, they were concerned about environmental changes and restrictions when proceeding with research during the COVID-19 pandemic, which was unfamiliar at the time. In the second survey, after overcoming environmental changes in the early stages of the pandemic, nursing researchers' concerns shifted to anxiety about the future, such as concerns about degree acquisition, employment and career advancement, and research results. The current study highlights various concerns among nursing researchers regarding research activities that have evolved over time during the pandemic. Academic societies must flexibly construct support measures for nursing researchers when a new infectious disease occurs. Such measures should be sensitive to the prevailing social circumstances and the evolving needs of researchers.

Self-Assembled Micro-Sized Hexagons Built from Short DNA in a Crowded Environment.

DNA programmable structures of various morphologies have attracted extensive attention due to their potential for materials science and biomedical applications. Here, we report the formation of micro-sized hexagons via assembly of only one pair of short double-stranded DNA in buffer-salt poly(ethylene glycol) solution. Each DNA strand had complementary bases with a two-base overhang. The procedure of heating and subsequent cooling of blunt-ended double-stranded DNA resulted in different assemblies. These results indicated that end-to-end adhesion at the terminals induced by complementary overhangs were required to construct the hexagonal DNA assemblies. The stable formation of the hexagons was highly dependent on heating temperature. In addition, concentration adjustments of DNA and poly(ethylene glycol) were essential. Circular dichroism spectral measurements and polarization microscopy observations indicated parallel alignment of double-stranded DNA in the hexagonal platelet. Self-assembled micro-sized hexagons composed of simple building blocks may have great potential for future biomedical device development.

Evaluation of DNA-mediated electron transfer using a hole-trapping nucleobase under crowded conditions.

The effects of a crowded environment on DNA-mediated electron transfer were evaluated using a pyrene-modified oligonucleotide containing a hole-trapping nucleobase in poly(ethylene glycol) mixed solutions. Rapid decompositions of hole-trapping bases in condensed and noncondensed DNA showed that more efficient electron transfer occurred under crowded conditions than in dilute solutions.

Backrest elevation in the semi-lateral position: Case of a sacral pressure ulcer with undermining formation.

Backrest elevation for a patient with a sacral-coccygeal pressure ulcer is necessary in certain situations, particularly to enable the patient to take meals. Deep pressure ulcers with undermining formations affect patients and create challenges for caregivers. The procedure of backrest elevation potentially worsens the pre-existing sacral-coccygeal pressure ulcers with undermining formations. Here, we report a Case of the clinical care of a patient using a simple approach for backrest elevation that minimizes additional injury to the existing sacral pressure ulcer covered with granulation tissue. In this case, we performed the backrest elevation in the semi-lateral position. After the backrest elevation, the patient was repositioned to the supine position to take a meal. The supine position was allowed at any time except during backrest elevation. The pressure ulcer of the patient improved rapidly using the combined treatment of our positioning approach and appropriate topical ointments. This practical approach may be effective for some patients as it potentially decreases the effect of shear force. The rationale for this approach can be explained by the mechanism of the undermining formation outlined in our experimental study using a pressure ulcer model mounted to a phantom. Although we used this position for patients with sacral-coccygeal pressure ulcers, this simple approach may also be considered for other patients based on our proposed pathogenesis of undermining formations.

Podoplanin is indispensable for cell motility and platelet-induced epithelial-to-mesenchymal transition-related gene expression in esophagus squamous carcinoma TE11A cells.

BACKGROUND: The transmembrane glycoprotein podoplanin (PDPN) is upregulated in some tumors and has gained attention as a malignant tumor biomarker. PDPN molecules have platelet aggregation-stimulating domains and, are therefore, suggested to play a role in tumor-induced platelet activation, which in turn triggers epithelial-to-mesenchymal transition (EMT) and enhances the invasive and metastatic activities of tumor cells. In addition, as forced PDPN expression itself can alter the propensity of certain tumor cells in favor of EMT and enhance their invasive ability, it is also considered to be involved in the cell signaling system. Nevertheless, underlying mechanisms of PDPN in tumor cell invasive ability as well as EMT induction, especially by platelets, are still not fully understood. METHODS: Subclonal TE11A cells were isolated from the human esophageal squamous carcinoma cell line TE11 and the effects of anti-PDPN neutralizing antibody as well as PDPN gene knockout on platelet-induced EMT-related gene expression were measured. Also, the effects of PDPN deficiency on cellular invasive ability and motility were assessed. RESULTS: PDPN-null cells were able to provoke platelet aggregation, suggesting that PDPN contribution to platelet activation in these cells is marginal. Nevertheless, expression of platelet-induced EMT-related genes, including vimentin, was impaired by PDPN-neutralizing antibody as well as PDPN deficiency, while their effects on TGF-ß-induced gene expression were marginal. Unexpectedly, PDPN gene ablation, at least in either allele, engendered spontaneous N-cadherin upregulation and claudin-1 downregulation. Despite these seemingly EMT-like alterations, PDPN deficiency impaired cellular motility and invasive ability even after TGF-ß-induced EMT induction. CONCLUSIONS: These results suggested that, while PDPN seems to function in favor of maintaining the epithelial state of this cell line, it is indispensable for platelet-mediated induction of particular mesenchymal marker genes as well as the potentiation of motility and invasion capacity.

Guanine damage by singlet oxygen from SYBR Green I in liquid crystalline DNA.

It is known that double-stranded DNA (dsDNA) turns into a liquid crystalline phase by the addition of a high concentration of polymer with salt. SYBR Green I (SG) is a well-known sensitive fluorescent stain for dsDNA, and is intercalated in liquid crystalline DNA. Formation of the liquid crystalline dsDNA-SG complex has been confirmed by CD spectral measurements, fluorescence spectral measurements and confocal fluorescence microscopy. SG in dsDNA was also used as a singlet oxygen generator. We conducted photoirradiation experiments using three kinds of 42-mer oligonucleotides with SG. The amount of guanine decomposition by selective irradiation of SG was analyzed using HPLC after digestion of dsDNA in each sample solution. We found that singlet oxygen produced in liquid crystalline DNA promoted guanine damage much more efficiently than in homogeneous solution.

The mechanism of persistent undermining of a sacral pressure ulcer: Experimental analyses using a deformable model and examination of skin mobility over different anatomical locations.

Undermining is an important issue in the treatment and care of deep pressure ulcers. The frequency of the undermining over different bony prominences varies. In particular, deep pressure ulcers over the sacrum exhibit undermining more frequently than those occurring over the heel. Although shear force has been suggested as a critical factor in undermining, the exact mechanism remains unclear due to ethical and technical reasons in clinical practice. To clarify this issue, a deformable model was constructed to recreate the physical and morphological properties of a pressure ulcer with persistent undermining. The model was constructed using urethane to recreate the physical properties of a pressure ulcer. To examine the clinical relevance of the model, mechanical properties of the skin and the model were measured using a durometer. The model was further mounted onto a phantom that was laid on a bed. Backrest elevation of the bed induced deformities in the urethane model, suggesting a mechanism of persistent undermining of the sacral pressure ulcer. Moreover, a simple palpation examination in elderly volunteers revealed that the skin over the sacrum was more mobile than the skin over the heel. Therefore, persistent undermining is likely caused by specific external forces and the characteristic skin mobility of the sacral region. These two different factors explain the frequent undermining that occurs in sacral pressure ulcers.

Magnetic Resonance Imaging Findings Related to Stroke Risk in Japanese Patients With Fabry Disease.

Background and Purpose- Stroke is the most frequent severe clinical event in patients with Fabry disease. We aimed to evaluate the cerebral magnetic resonance imaging findings of patients with Fabry disease and assess their association with future stroke risk after enzyme replacement therapy (ERT) induction. Methods- We retrospectively reviewed the magnetic resonance imaging findings of 25 patients with Fabry disease. Of these, 12 adult patients without prior stroke or ERT were enrolled to evaluate the association between baseline magnetic resonance imaging findings and stroke occurrence after ERT initiation. We assessed white matter hyperintensities, periventricular hyperintensities, and basilar artery diameter as neuroimaging markers. Results- The mean age of participants was 38.8±16.8 years. Fourteen patients (56.0%) were women. White matter hyperintensities and periventricular hyperintensities were observed in 15 and 13 cases, respectively. The mean basilar artery diameter was 3.2±0.55 mm. Two patients demonstrated old infarct lesions. Three of 12 patients in whom ERT was initiated had symptomatic stroke after baseline magnetic resonance imaging. Of these, 2 developed stroke within 6-month post-ERT initiation. The baseline basilar artery diameter was larger in patients who developed stroke than in those who did not (4.0±0.40 versus 2.9±0.49 mm, P=0.02). Conclusions- Basilar artery diameter may be associated with stroke risk after ERT initiation among Japanese patients with Fabry disease.

The effect of spermidine on guanine decomposition via photoinduced electron transfer in DNA.

Spermidine, a trivalent organic cation, induced DNA structural changes and suppressed guanine photooxidative decomposition via electron transfer through pyrene-modified DNA. On the other hand, adding higher concentrations of spermidine resulted in DNA condensation. The efficiency of guanine decomposition in condensed pyrene-modified DNA was promoted remarkably.

Anterior Cerebral Artery Dissection in a Patient With Autosomal Dominant Polycystic Kidney Disease.

Intracranial artery dissection secondary to autosomal dominant polycystic kidney disease is far less common than cerebral aneurysm. A 55-year-old man presented a sudden onset of headache and disturbed consciousness caused by ischemic stroke in the bilateral frontal lobes with minor subarachnoid hemorrhage. The bilateral anterior cerebral arteries were firstly occluded and re-perfused with irregular narrowing and dilation in 3 days after stroke onset, indicating dissection. He was diagnosed with autosomal dominant polycystic kidney disease by abdominal CT findings and by his family history though his renal function was almost normal. Dissection in the anterior cerebral artery has not been reported previously, while some cases with dissection in the vertebral and extracranial arteries were reported in autosomal dominant polycystic kidney disease. His family also had a history of aortic dissection and subarachnoid hemorrhage. Intracranial artery dissection may be a manifestation of systemic arteriopathy with familial clustering in autosomal dominant polycystic kidney disease. Strict antihypertensive treatment is needed in these cases.

Correction to: Acquisition of Stop Consonants in Mandarin-Japanese Early Bilingual Children.

The original version of the article unfortunately contained an error in Acknowledgement section.

Acquisition of Stop Consonants in Mandarin-Japanese Early Bilingual Children.

This study examines the nature of stop accuracy and substitute patterns of word-initial Japanese and Mandarin stops produced by Mandarin-Japanese bilingual children. The purpose of the study is to understand phonological development in bilinguals. The sample consists of 36 bilingual children between the ages of three and six, who simultaneously acquired Japanese and Mandarin from birth. The results were as follows: (1) most of the bilingual children were able to produce Mandarin and Japanese stops by the age of three and the accuracy of the target stops were found to develop with age; (2) the age of developing the target consonants is slightly different in the two languages; (3) substitution patterns observed in each language reveals a mixture of child-specific patterns, language specific systems and language influence as well as individual differences. These findings indicate that Mandarin-Japanese bilingual children possess a unique phonological development system, which is a monolinguallike pattern with cross-linguistic interaction. These results constitute a new body of descriptive reference materials documenting the phonological development of bilingual children for speech therapists or pathologists.

Attenuation of guanine oxidation via DNA-mediated electron transfer in a crowded environment using small cosolutes.

Guanine oxidation induced by photoirradiation on a pyrene-modified oligonucleotide was investigated under molecular crowding using small cosolutes such as glycerol. The efficiency of guanine photooxidation was suppressed in accordance with the increase in the concentration of glycerol. The results of photooxidation experiments using fully matched and mismatched DNA showed that guanine decomposition was mainly caused by DNA-mediated electron transfer (ET) in glycerol mixed solutions, as well as in diluted aqueous buffer solutions. Multiple factors can contribute to the suppression of guanine oxidation in crowded environments. However, our experimental results indicated that the attenuation of the DNA-mediated ET process suppressed guanine oxidation. On the other hand, experiments using ethylene glycol showed that the guanine decomposition efficiency varies depending on the surrounding solvent. These results suggested that changes in the characteristics of the surrounding medium affect the DNA fluctuation, dominating DNA-mediated ET.

Using the Acropora digitifera genome to understand coral responses to environmental change.

Despite the enormous ecological and economic importance of coral reefs, the keystone organisms in their establishment, the scleractinian corals, increasingly face a range of anthropogenic challenges including ocean acidification and seawater temperature rise. To understand better the molecular mechanisms underlying coral biology, here we decoded the approximately 420-megabase genome of Acropora digitifera using next-generation sequencing technology. This genome contains approximately 23,700 gene models. Molecular phylogenetics indicate that the coral and the sea anemone Nematostella vectensis diverged approximately 500 million years ago, considerably earlier than the time over which modern corals are represented in the fossil record (∼240 million years ago). Despite the long evolutionary history of the endosymbiosis, no evidence was found for horizontal transfer of genes from symbiont to host. However, unlike several other corals, Acropora seems to lack an enzyme essential for cysteine biosynthesis, implying dependency of this coral on its symbionts for this amino acid. Corals inhabit environments where they are frequently exposed to high levels of solar radiation, and analysis of the Acropora genome data indicates that the coral host can independently carry out de novo synthesis of mycosporine-like amino acids, which are potent ultraviolet-protective compounds. In addition, the coral innate immunity repertoire is notably more complex than that of the sea anemone, indicating that some of these genes may have roles in symbiosis or coloniality. A number of genes with putative roles in calcification were identified, and several of these are restricted to corals. The coral genome provides a platform for understanding the molecular basis of symbiosis and responses to environmental changes.

Wound fixation for pressure ulcers: a new therapeutic concept based on the physical properties of wounds.

A pressure ulcer is defined as damage to skin and other tissues over a bony prominence caused by excess pressure. Deep pressure ulcers that develop over specific bony prominences often exhibit wound deformity, defined as a change in the 3-dimensional shape of the wound. Subsequently, the wound deformity can result in undermining formation, which is a characteristic of deep pressure ulcers. However, to date, a concept with respect to alleviating wound deformity has yet to be defined and described. To clarify the issue, we propose a new concept called "wound fixation" based on the physical properties of deep pressure ulcers with wound deformity. Wound fixation is defined here as the alleviation of wound deformity by exogenous materials. The wound fixation methods are classified as traction, anchor, and insertion based on the relation between the wound and action point by the exogenous materials. A retrospective survey of a case series showed that wound fixation was preferentially used for deep pressure ulcers at specific locations such as the sacrum, coccyx, and greater trochanter. Moreover, the methods of wound fixation were dependent on the pressure ulcer location. In conclusion, our new concept of wound fixation will be useful for the practical treatment and care of pressure ulcers. Further discussion and validation by other experts will be required to establish this concept.

Association between interleukin-6 levels and first-ever cerebrovascular events in patients with vascular risk factors.

OBJECTIVE: The objective of this study was to examine the association of inflammatory markers with risk of first-ever cerebrovascular events (CVEs), while simultaneously evaluating subclinical vascular disease. METHODS AND RESULTS: We enrolled 464 outpatients who had vascular risk factors without any preexisting cardiovascular disease. We examined the presence of silent lacunar infarction (SLI) by magnetic resonance imaging; carotid intima-media thickness by ultrasound; and measured high-sensitivity C-reactive protein, interleukin (IL)-6, and IL-18 at baseline, and assessed their associations with CVEs using Cox proportional hazards models of 4.8±2.6 years follow-up. We further calculated measures of reclassification and discrimination. In age- and sex-adjusted analysis, IL-6, but neither high-sensitivity C-reactive protein nor IL-18, was associated with CVEs. The association remained significant after adjustment for conventional risk factors, intima-media thickness, and SLI (hazard ratios: 1.80, per 1-SD increase in log IL-6, P=0.03). Compared with the patients with below median IL-6 without SLI, those with above median IL-6 and SLI had a higher risk of CVEs (hazard ratios: 4.14, P=0.0014). The combination of IL-6 and SLI resulted in the net reclassification improvement of 14.3% (P=0.04), and the integrated discrimination improvement gain of 2.1% (P=0.05). CONCLUSIONS: IL-6 levels were independently associated with CVEs and could improve reclassification in those with SLI.

Basilar artery diameter is an independent predictor of incident cardiovascular events.

OBJECTIVE: Basilar arterial (BA) dolichoectasia is associated with cerebral small-vessel disease and stroke. However, the association between moderate dilation of the BA and cerebral small-vessel disease or subsequent cardiovascular events remains unclear. This study aims to clarify the factors related to BA diameter and to clarify whether the BA diameter is an independent predictor of cardiovascular events. APPROACH AND RESULTS: The study subjects comprised 493 outpatients with atherosclerotic risk factors. BA diameter, lacunar infarct, severity of deep white matter hyperintensities, and intracranial steno-occlusive lesions were assessed with MRI and magnetic resonance angiography. Then, we prospectively evaluated the association between BA diameter and cardiovascular events. The BA diameter ranged from 1.1 to 5.2 mm, and only 0.8% of the patients had dolichoectasia. Male sex, the presence of lacunar infarcts, the severity of deep white matter hyperintensities, the fetal-type variation of the circle of Willis, and intracranial steno-occlusive lesions were independently associated with BA diameter. In the mean follow-up of 6.0 years, 91 patients developed cardiovascular events. BA diameter was independently associated with total cardiovascular events after adjusting for age, sex, and conventional risk factors (hazard ratio, 1.55 per 1 mm increase in BA diameter; P=0.009). CONCLUSIONS: Increased BA diameter within the normal range is related to both large-vessel disease and cerebral small-vessel disease, and it could be a new predictor of cardiovascular events.

Probing a coral genome for components of the photoprotective scytonemin biosynthetic pathway and the 2-aminoethylphosphonate pathway.

Genome sequences of the reef-building coral, Acropora digitifera, have been decoded. Acropora inhabits an environment with intense ultraviolet exposure and hosts the photosynthetic endosymbiont, Symbiodinium. Acropora homologs of all four genes necessary for biosynthesis of the photoprotective cyanobacterial compound, shinorine, are present. Among metazoans, these genes are found only in anthozoans. To gain further evolutionary insights into biosynthesis of photoprotective compounds and associated coral proteins, we surveyed the Acropora genome for 18 clustered genes involved in cyanobacterial synthesis of the anti-UV compound, scytonemin, even though it had not previously been detected in corals. We identified candidates for only 6 of the 18 genes, including tyrP, scyA, and scyB. Therefore, it does not appear that Acropora digitifera can synthesize scytonemin independently. On the other hand, molecular phylogenetic analysis showed that one tyrosinase gene is an ortholog of vertebrate tyrosinase genes and that the coral homologs, scyA and scyB, are similar to bacterial metabolic genes, phosphonopyruvate (ppyr) decarboxylase and glutamate dehydrogenase (GDH), respectively. Further genomic searches for ppyr gene-related biosynthetic components indicate that the coral possesses a metabolic pathway similar to the bacterial 2-aminoethylphosphonate (AEP) biosynthetic pathway. The results suggest that de novo synthesis of carbon-phosphorus compounds is performed in corals.

Enhancement of fluorescence quenching and exciplex formation in DNA major groove by double incorporation of modified fluorescent deoxyuridines.

5-(1-Naphthalenylethynyl)-2'-deoxyuridine ((N)U) and 5-[(4-cyano-1-naphthalenyl)ethynyl]-2'-deoxyuridine ((CN)U) were synthesized and incorporated into oligodeoxynucleotides. Fluorescence emissions of modified duplexes containing double (N)U were efficiently quenched depending upon the sequence pattern of the naphthalenes in DNA major groove, as compared to the duplex possessing single (N)U. When one of the naphthalene moieties has a cyano substituent, the exciplex emission from the chromophores in DNA major groove was observed at longer wavelength.