Predominant accumulation of a 3-hydroxy-γ-decalactone in the male rectal gland complex of the Japanese orange fly, Bactrocera tsuneonis

The Japanese orange fly, Bactrocera tsuneonis, infests various citrus crops. While male pheromone components accumulated in the rectal glands are well characterized for Bactrocera, but information regarding the chemical factors involved in the life cycles of B. tsuneonis remains scarce. Herein, several volatile chemicals including a γ-decalactone, (3R, 4R)-3-hydroxy-4-decanolide [(3R, 4R)-HD], were identified as major components, along with acetamide and spiroketals as minor components in the rectal gland complexes of male B. tsuneonis flies. The lactone (3R, 4R)-HD was also identified in female rectal gland complexes. The amount of this compound in mature males was significantly higher than those observed in females and immature males. The lactone (3R, 4R)-HD was detected in flies fed with sucrose only, indicating that this lactone is not derived from dietary sources during adulthood, but biosynthesized in vivo. The predominant accumulation of (3R, 4R)-HD in mature males also suggests a possible role in reproductive behavior

LACTATE/MCT4/GPR81 AXIS IN BONE PAIN OF BREAST CANCER

Breast cancer (BC) bone metastasis causes bone pain (BP), which detrimentally damages the quality of life and outcome of patients with BC. However, the mechanism of BC‑BP is poorly understood, and effective treatments are limited. The present study demonstrated a novel mechanism of BC‑BP using a mouse model of bone pain, in which mouse (EO771) and human (MDA‑MB‑231) BC cells were injected in the bone marrow cavity of tibiae. Western blot analysis using sensory nerves, in vivo assessment of cancer pain and in vitro calcium flux analysis were performed. These mice developed progressive BC‑BP in tibiae in conjunction with an upregulation of phosphorylated pERK1/2 and cAMP‑response element‑binding protein (pCREB), which are molecular indicators of neuron excitation, in the dorsal root ganglia (DRG) of sensory nerves. Importantly, mice injected with BC cells, in which the expression of the lactic acid transporter monocarboxylate transporter 4 (MCT4) was silenced, exhibited decreased BC‑BP with downregulated expression of pERK1/2 and pCREB in the DRG and reduced circulating levels of lactate compared with mice injected with parental BC cells. Further, silencing of the cell‑surface orphan receptor for lactate, G protein‑coupled receptor 81 (GPR81), in the F11 sensory neuron cells decreased lactate‑promoted upregulation of pERK1/2 and Ca2+ influx, suggesting that the sensory neuro excitation was inhibited. These results suggested that lactate released from BC cells via MCT4 induced BC‑BP through the activation of GPR81 of sensory neurons. In conclusion, the activation of GPR81 of sensory neurons by lactate released via MCT4 from BC was demonstrated to contribute to the induction of BC‑BP, and disruption of the interactions among lactate, MCT4 and GPR81 may be a novel approach to control BC‑BP

日本における保険診療全透析患者追跡と死亡数の現状

Background: The survival rate of chronic dialysis patients in Japan remains the highest worldwide, so there is value in presenting Japan's situation internationally. We examined whether aggregate figures on dialysis patients in the National Database of Health Insurance Claims and Special Health Checkups of Japan (NDB), which contains data on insured procedures of approximately 100 million Japanese residents, complement corresponding figures in the Japanese Society for Dialysis Therapy Renal Data Registry (JRDR). Methods: Subjects were patients with medical fee points for dialysis recorded in the NDB during 2014-2018. We analyzed annual numbers of dialysis cases, newly initiated dialysis cases- and deaths. Results: Compared with the JRDR, the NDB had about 6-7% fewer dialysis cases but a similar number of newly initiated dialysis cases. In the NDB, the number of deaths was about 6-10% lower, and the number of hemodialysis cases was lower, while that of peritoneal dialysis cases was higher. The cumulative survival rate at dialysis initiation was approximately 6 percentage points lower in the NDB than in the JRDR, indicating that some patients die at dialysis initiation. Cumulative survival rate by age group was roughly the same between the NDB and JRDR in both sexes. Conclusion: The use of the NDB enabled us to aggregate data of dialysis patients. With the definition of dialysis patients used in this study, analyses of concomitant medications, comorbidities, surgeries, and therapies will become possible, which will be useful in many future studies.博士（医学）・甲第818号・令和4年3月15日© 2021. The Author(s).　Open Access This article is licensed under a Creative Commons Attri bution 4.0 International License, which permits use, sharing, adapta tion, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/

Three-dimensional microvascular network reconstruction from in vivo images with adaptation of the regional inhomogeneity in the signal-to-noise ratio

ObjectiveQuantification of angiographic images with two-photon laser scanning fluorescence microscopy (2PLSM) relies on proper segmentation of the vascular images. However, the images contain inhomogeneities in the signal-to-noise ratio (SNR) arising from regional effects of light scattering and absorption. The present study developed a semiautomated quantification method for volume images of 2PLSM angiography by adjusting the binarization threshold according to local SNR along the vessel centerlines.MethodsA phantom model made with fluorescent microbeads was used to incorporate a region-dependent binarization threshold.ResultsThe recommended SNR for imaging was found to be 4.2–10.6 that provide the true size of imaged objects if the binarization threshold was fixed at 50% of SNR. However, angiographic images in the mouse cortex showed variable SNR up to 45 over the depths. To minimize the errors caused by variable SNR and a spatial extent of the imaged objects in an axial direction, the microvascular networks were three-dimensionally reconstructed based on the cross-sectional diameters measured along the vessel centerline from the XY-plane images with adapted binarization threshold. The arterial volume was relatively constant over depths of 0–500 µm, and the capillary volume (1.7% relative to the scanned volume) showed the larger volumes than the artery (0.8%) and vein (0.6%).ConclusionsThe present methods allow consistent segmentation of microvasculature by adapting the local inhomogeneity in the SNR, which will be useful for quantitative comparison of the microvascular networks, such as under disease conditions where SNR in the 2PLSM images varies over space and time

sj-pdf-1-jcb-10.1177_0271678X221125743 - Supplemental material for Spatiotemporal analysis of blood plasma and blood cell flow fluctuations of cerebral microcirculation in anesthetized rats

Supplemental material, sj-pdf-1-jcb-10.1177_0271678X221125743 for Spatiotemporal analysis of blood plasma and blood cell flow fluctuations of cerebral microcirculation in anesthetized rats by Tomoya Niizawa, Ruka Sakuraba, Tomoya Kusaka, Yuika Kurihara, Takuma Sugashi, Hiroshi Kawaguchi, Iwao Kanno and Kazuto Masamoto in Journal of Cerebral Blood Flow & Metabolism</p

HIV-1 Suppressive Sequences Are Modulated by Rev Transport of Unspliced RNA and Are Required for Efficient HIV-1 Production

<div><p>The unspliced human immunodeficiency virus type 1 (HIV-1) RNAs are translated as Gag and Gag-Pol polyproteins or packaged as genomes into viral particles. Efficient translation is necessary before the transition to produce infective virions. The viral protein Rev exports all intron-containing viral RNAs; however, it also appears to enhance translation. Cellular microRNAs target cellular and viral mRNAs to silence their translation and enrich them at discrete cytoplasmic loci that overlap with the putative interim site of Gag and the genome. Here, we analyzed how Rev-mediated transport and the splicing status of the mRNA influenced the silencing status imposed by microRNA. Through identification and mutational analysis of the silencing sites in the HIV-1 genome, we elucidated the effect of silencing on virus production. <em>Renilla</em> luciferase mRNA, which contains a let-7 targeting site in its 3′ untranslated region, was mediated when it was transported by Rev and not spliced, but it was either not mediated when it was spliced even in a partial way or it was Rev-independent. The silencing sites in the <em>pol</em> and <em>env</em>-<em>nef</em> regions of the HIV-1 genome, which were repressed in T cells and other cell lines, were Drosha-dependent and could also be modulated by Rev in an unspliced state. Mutant viruses that contained genomic mutations that reflect alterations to show more derepressive effects in the 3′ untranslated region of the <em>Renilla</em> luciferase gene replicated more slowly than wild-type virus. These findings yield insights into the HIV-1 silencing sites that might allow the genome to avoid translational machinery and that might be utilized in coordinating virus production during initial virus replication. However, the function of Rev to modulate the silencing sites of unspliced RNAs would be advantageous for the efficient translation that is required to support protein production prior to viral packaging and particle production.</p> </div

The effect of Rev-dependent export for Let-7 family members and suppressive sequences within HIV-1 in Jurkat cells.

<p>(A) The secondary structure of the miRNA/mRNA duplex of let-7 family members targeting the Bulge sequence was predicted using the RNA-hybrid program. (B) The relative RNA expression levels in the let-7 family members from Jurkat cells were calculated following normalization to the internal control RUN6B. (C) The silencing of RNAs containing the let-7 targeting-sequences in Jurkat cells. (D) The effects of the Bulge and BulgeMut sequences on RNA export by Rev-HA in Jurkat cells. The blue arrow points to the Bulge-containing constructs and the black arrow points to the BulgeMut-containing vectors. Three independent experiments were performed. **P<0.005, *P<0.05. (E) The effect of Rev-mediated export on RNAs in which the suppressive sequences identified in the <i>pol</i> (element 1 in <b>Fig. 5A</b>) and <i>env</i>-<i>nef</i> (element 15 in <b>Fig. 5A</b>) regions were inserted into the <i>Rluc</i> 3′ UTR was assessed in Jurkat cells (black bars, blue arrows). The corresponding vectors with mutations in the repressive sites (lattice and blue bars, black arrows) were also assessed as well (mutation 1-2m-2 in the <i>pol</i> region and 15-3m-2 in the <i>env</i>-<i>nef</i> region; <b>Fig. 5B–G</b>). The bar patterns correspond to the mutational patterns shown in <b>Fig. 5</b>. The <i>Renilla</i>/firefly luciferase value was assessed in each graph. The empty vector C was used as a control, and the results are presented as the mean ± S.D. as a percentage of the control. “pcDNA” denotes the pcDNA3.1(+) plasmid.</p

The effects of mutations in suppressive sequences in the <i>pol</i> and <i>env</i>-<i>nef</i> regions.

<p>(A) The <i>pol</i> and <i>env</i>-<i>nef</i> regions of pNL4-3 are illustrated. The <i>cis</i>-acting instability elements (INS) are also illustrated below the genome. The arrows indicate the mutated sites in the <i>pol</i> (sites “a” and “b”) and <i>env</i>-<i>nef</i> (sites “c”, “d” and “e”) regions. The black (>40%) and gray (40–20%) bars represent the repressed region. The number and bar pattern corresponds to the graph below. (B) The effects of mutations introduced into the repressive sequences in the <i>pol</i> region (sites “a” and “b”) were assessed in Jurkat cells. The “1 m” indicates the vector that has the mutated sequence introduced into the repressive site. (C) Mutational assay of the suppressive sequences (sites “c” and “d”) in the <i>env</i>-<i>nef</i> region. (D) Mutational assay of the suppressive sequences (site “e”) in the <i>nef</i> region. The <i>Rluc</i> activity was normalized to the firefly luciferase activity, and the data shown are the mean ± S.D. of the percentage of the activity of the empty psiCHECK-2 vector (psiCHECK). (E) Pattern mutations and the effects of each dissected sequence in the <i>pol</i> (sites “a” and “b”) and <i>env</i>-<i>nef</i> (sites “c”, “d” and “e”) regions. The gray characters represent unchanged residues. Asterisks denote mutations that did not change any amino acids (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0051393#pone.0051393.s010" target="_blank">Table S1</a> for details).</p

The effects of combining multiple mutations in the <i>pol</i> and <i>env</i>-<i>nef</i> silencing regions.

<p>(A) The <i>pol</i> and <i>env</i>-<i>nef</i> regions of pNL4-3 and the position of the <i>cis</i>-acting instability elements (INS) are illustrated. The arrows indicate the mutated sites in the <i>pol</i> (sites “a” and “b”) and <i>env</i>-<i>nef</i> (sites “c”, “d” and “e”) regions. The number and bar pattern corresponds to the graph below. (B) The effects of multiple mutations in the <i>pol</i> region (sites “a” and “b”; elements 1 and 7) were assessed in Jurkat cells. Six independent experiments were performed. The bar patterns correspond to the mutational patterns shown in (F). The “2 m” indicates that the vector has two mutation sites in the repressive sequence. (C) The effects of multiple mutations in the <i>pol</i> region were assessed in M4C8 cells. (D) The effects of multiple mutations introduced into the <i>env</i>-<i>nef</i> region (sites “c”, “d” and “e”; elements 15 and 16) were assessed in Jurkat cells. Four independent experiments were performed. The bar patterns correspond to the mutational patterns shown in (G). The “1 m”, “2 m” and “3 m” indicate that the vector has one, two and three mutation sites individually. The red and blue bars indicate the more derepressed mutational patterns. (E) The effects of multiple mutations introduced into the <i>env</i>-<i>nef</i> region were assessed in M4C8 cells. The psiCHECK was used as a control, and the results are expressed as the mean ± S.D. as a percentage of the control. ***P<0.001, **P<0.005 and *P<0.05. (F) Combinations of mutated patterns in the <i>pol</i> region and the bar patterns in graph B and C. (G) Combinations of mutated patterns in the <i>env</i>-<i>nef</i> region and the bar patterns in graph D and E. The gray characters represent unchanged residues. The asterisk denotes the mutational pattern used for the generation of mutant viruses.</p