Heat stress memory differentially regulates the expression of nitrogen transporter genes in the filamentous red alga 'Bangia' sp. ESS1.

Introduction: To withstand high temperatures that would be lethal to a plant in the naïve state, land plants must establish heat stress memory. The acquisition of heat stress tolerance via heat stress memory in algae has only been observed in the red alga 'Bangia' sp. ESS1. Methods: In this study, we further evaluated the intrinsic ability of this alga to establish heat stress memory by monitoring hydrogen peroxide (H2O2) production and examining the relationship between heat stress memory and the expression of genes encoding nitrogen transporters, since heat stress generally reduces nitrogen absorption. Next, genes encoding nitrogen transporters were selected from our unpublished transcriptome data of 'Bangia' sp. ESS1. Results: We observed a reduction in H2O2 content when heat stress memory was established in the alga. In addition, six ammonium transporter genes, a single-copy nitrate transporter gene and two urea transporter genes were identified. Two of these nitrogen transporter genes were induced by heat stress but not by heat stress memory, two genes showed heat stress memory-dependent expression, and one gene was induced by both treatments. Heat stress memory therefore differentially regulated the expression of the nitrogen transporter genes by reducing heat stress-inducible gene expression and inducing heat stress memory-dependent gene expression. Discussion: These findings point to the functional diversity of nitrogen transporter genes, which play different roles under various heat stress conditions. The characteristic effects of heat stress memory on the expression of individual nitrogen transporter genes might represent an indispensable strategy for reducing the threshold of sensitivity to recurrent high-temperature conditions and for maintaining nitrogen absorption under such conditions in 'Bangia' sp. ESS1.

A radiation-related second primary gastrointestinal stromal tumor in the bladder.

A 69-year-old man underwent 78 Gy/39 Fr of intensity-modulated radiation therapy for prostate cancer. Seven years after radiotherapy, a nonpapillary bladder tumor was identified. Transurethral resection of the bladder tumor was performed, and the pathological diagnosis was spindle cell sarcoma. Immunostaining revealed KIT-, DOG1++, CD34-, Actin++, Cytokeratin-, Desmin-, S100 protein-, and Vimentin++. No tumor recurrence was observed until 17 months after tumor resection. DOG1 is strongly and specifically expressed in gastrointestinal stromal tumors. This was a rare case of bladder gastrointestinal stromal tumor as a radiation-related second primary tumor.

Hypomethylation of a CpG Site in the CpG Island of the Aquaporin 1 Gene May Be Involved in the Formation of Adult-Onset Non-communicating Hydrocele Testis.

Background Water channel aquaporin 1 (AQP1) protein expression is enhanced in the tunica vaginalis of patients with adult-onset non-communicating hydrocele testis and may contribute to the development of non-communicating hydrocele testis. We performed genetic and epigenetic analyses of the AQP1 gene in the tunica vaginalis of patients with adult-onset non-communicating hydrocele testis to elucidate the cause of enhanced AQP1 protein expression. Methodology The genotype was determined for Tag single-nucleotide polymorphisms (SNPs) representing the AQP1 gene and SNPs in the 5'-upstream region of the AQP1 gene. Then, by performing association analysis, the applicability of various genetic models was investigated for each SNP. Moreover, the methylation rate of CpG sites was examined for the CpG island related to the AQP1 gene. Results There was no significant association between each SNP and hydrocele testis for any of the genetic models. The average methylation rate of the 17 CpG sites evaluated was not significantly different between controls and hydrocele testis, but the methylation rate was lower in hydrocele testis than in controls at one CpG site. Conclusions There was a significant decrease in the methylation rate at one of the CpG sites in the CpG island associated with the AQP1 gene in the tunica vaginalis of patients with non-communicating hydrocele testis. This may increase AQP1 protein expression and contribute to the formation of hydrocele testis. SNPs related to the AQP1 gene were not associated with hydrocele testis.

Structural Analysis of the Colony-Stimulating Factor 3 Gene of Granulocyte Colony-Stimulating Factor-Producing Urothelial Cancer.

Background Granulocyte colony-stimulating factor (G-CSF) is a member of the CSF family of glycoproteins that regulate the proliferation, differentiation, and mobilization of neutrophils. G-CSF-producing malignant cancers have been reported to occur in various organs and are mostly associated with poor clinical prognosis. Here, we analyzed the structure of the CSF3 gene encoding the G-CSF protein to delineate the mechanism of G-CSF production by the cancer cells. Methodology Two cases of G-CSF-producing urothelial cancers and three cases of G-CSF-nonproducing bladder cancers were enrolled for genetic analysis. Results In one case of G-CSF-producing bladder cancer, six somatic mutations were detected in the 5'- upstream region of the CSF3 gene. No somatic mutations in the CSF3 gene were detected in another case of G-CSF-producing renal pelvic cancer and G-CSF-nonproducing bladder cancers. Copy numbers of the CSF3 gene were not increased in G-CSF-producing urothelial cancers. Conclusions Somatic mutations in the 5'- upstream region of the CSF3 gene may cause G-CSF protein overproduction.

Membrane Fluidization Governs the Coordinated Heat-Inducible Expression of Nucleus- and Plastid Genome-Encoded Heat Shock Protein 70 Genes in the Marine Red Alga Neopyropia yezoensis.

Heat shock protein 70 (HSP70) is an evolutionarily conserved protein chaperone in prokaryotic and eukaryotic organisms. This family is involved in the maintenance of physiological homeostasis by ensuring the proper folding and refolding of proteins. The HSP70 family in terrestrial plants can be divided into cytoplasm, endoplasmic reticulum (ER)-, mitochondrion (MT)-, and chloroplast (CP)-localized HSP70 subfamilies. In the marine red alga Neopyropia yezoensis, the heat-inducible expression of two cytoplasmic HSP70 genes has been characterized; however, little is known about the presence of other HSP70 subfamilies and their expression profiles under heat stress conditions. Here, we identified genes encoding one MT and two ER HSP70 proteins and confirmed their heat-inducible expression at 25 °C. In addition, we determined that membrane fluidization directs gene expression for the ER-, MT-, and CP-localized HSP70 proteins as with cytoplasmic HSP70s. The gene for the CP-localized HSP70 is carried by the chloroplast genome; thus, our results indicate that membrane fluidization is a trigger for the coordinated heat-driven induction of HSP70 genes harbored by the nuclear and plastid genomes in N. yezoensis. We propose this mechanism as a unique regulatory system common in the Bangiales, in which the CP-localized HSP70 is usually encoded in the chloroplast genome.

Combination therapy with anti-programmed cell death 1 antibody plus angiokinase inhibitor exerts synergistic antitumor effect against malignant mesothelioma via tumor microenvironment modulation.

Malignant pleural mesothelioma (MPM) is an asbestos-related fatal malignant neoplasm. Although there has been no reliable chemotherapeutic regimen other than combination therapy of cisplatin and pemetrexed for two decades, combination of ipilimumab plus nivolumab brought about a better outcome in patients with MPM. Thus, cancer immunotherapy using immune checkpoint inhibitor (ICI) is expected to play a central role in the treatment of MPM. To maximize the antitumor effect of ICI, we evaluated whether nintedanib, an antiangiogenic agent, could augment the antitumor effect of anti-programmed cell death 1 (PD-1) antibody (Ab). Although nintedanib could not inhibit the proliferation of mesothelioma cells in vitro, it significantly suppressed the growth of mesothelioma allografts in mice. Moreover, combination therapy with anti-PD-1 Ab plus nintedanib reduced tumor burden more dramatically compared with nintedanib monotherapy via inducing remarkable necrosis in MPM allografts. Nintedanib did not promote the infiltration of CD8+ T cells within the tumor when used alone or in combination with anti-PD-1 Ab but it independently decreased the infiltration of tumor-associated macrophages (TAMs). Moreover, immunohistochemical analysis and ex vivo study using bone marrow-derived macrophages (BMDMs) showed that nintedanib could polarize TAMs from M2 to M1 phenotype. These results indicated that nintedanib had a potential to suppress protumor activity of TAMs both numerically and functionally. On the other hand, ex vivo study revealed that nintedanib upregulated the expression of PD-1 and PD-ligand 1 (PD-L1) in BMDMs and mesothelioma cells, respectively, and exhibited the impairment of phagocytic activity of BMDMs against mesothelioma cells. Co-administration of anti-PD-1 Ab may reactivate phagocytic activity of BMDMs by disrupting nintedanib-induced immunosuppressive signal via binding between PD-1 on BMDMs and PD-L1 on mesothelioma cells. Collectively, combination therapy of anti-PD-1 Ab plus nintedanib enhances the antitumor activity compared with respective monotherapy and can become a novel therapeutic option for patients with MPM.

Life cycle and reproduction dynamics of Bangiales in response to environmental stresses.

Red algae of the order Bangiales are notable for exhibiting flexible promotion of sexual and asexual reproductive processes by environmental stresses. This flexibility indicates that a trade-off between vegetative growth and reproduction occurs in response to environmental stresses that influence the timing of phase transition within the life cycle. Despite their high phylogenetic divergence, both filamentous and foliose red alga in the order Bangiales exhibit a haploid-diploid life cycle, with a haploid leafy or filamentous gametophyte (thallus) and a diploid filamentous sporophyte (conchocelis). Unlike haploid-diploid life cycles in other orders, the gametophyte in Bangiales is generated independently of meiosis; the regulation of this generation transition is not fully understood. Based on transcriptome and gene expression analyses, the originally proposed biphasic model for alternation of generations in Bangiales was recently updated to include a third stage. Along with the haploid gametophyte and diploid sporophyte, the triphasic framework recognizes a diploid conchosporophyte-a conchosporangium generated on the conchocelis-phase and previously considered to be part of the sporophyte. In addition to this sexual life cycle, some Bangiales species have an asexual life cycle in which vegetative cells of the thallus develop into haploid asexual spores, which are then released from the thallus to produce clonal thalli. Here, we summarize the current knowledge of the triphasic life cycle and life cycle trade-off in Neopyropia yezoensis and 'Bangia' sp. as model organisms for the Bangiales.

Successful Intrasaccular Coil Embolization of Mycotic Pulmonary Artery Pseudoaneurysm Using the Balloon Remodeling Technique.

A 66-year-old man with ventricular septal defect was hospitalized for fever, dyspnea, and hemoptysis. Infectious endocarditis by Streptococcus parasanguinis was suspected. At 3 weeks after admission, massive hemoptysis suddenly occurred. Contrast-enhanced computed tomography showed a 2-cm aneurysmal formation in the right lower pulmonary artery. Rupture of a mycotic pulmonary artery pseudoaneurysm was diagnosed. Pulmonary angiography showed a wide-necked pseudoaneurysm at the trifurcation of the anterior, lateral, and posterior basal segmental arteries of the right lower lobe. Intrasaccular coil embolization with the balloon remodeling technique was successfully performed, preserving residual pulmonary function distal to the pseudoaneurysm.

Auxin Regulates Apical Stem Cell Regeneration and Tip Growth in the Marine Red Alga Neopyropia yezoensis.

The red alga Neopyropia yezoensis undergoes polarized elongation and asymmetrical cell division of the apical stem cell during tip growth in filamentous generations of its life cycle: the conchocelis and conchosporangium. Side branches are also produced via tip growth, a process involving the regeneration and asymmetrical division of the apical stem cell. Here, we demonstrate that auxin plays a crucial role in these processes by using the auxin antagonist 2-(1H-Indol-3-yl)-4-oxo-4-phenyl-butyric acid (PEO-IAA), which specifically blocks the activity of the auxin receptor TRANSPORT INHIBITOR RESPONSE1 (TIR1) in land plants. PEO-IAA repressed both the regeneration and polarized tip growth of the apical stem cell in single-celled conchocelis; this phenomenon was reversed by treatment with the auxin indole-3-acetic acid (IAA). In addition, tip growth of the conchosporangium was accelerated by IAA treatment but repressed by PEO-IAA treatment. These findings indicate that auxin regulates polarized tip cell growth and that an auxin receptor-like protein is present in N. yezoensis. The sensitivity to different 5-alkoxy-IAA analogs differs considerably between N. yezoensis and Arabidopsis thaliana. N. yezoensis lacks a gene encoding TIR1, indicating that its auxin receptor-like protein differs from the auxin receptor of terrestrial plants. These findings shed light on auxin-induced mechanisms and the regulation of tip growth in plants.

Membrane-Fluidization-Dependent and -Independent Pathways Are Involved in Heat-Stress-Inducible Gene Expression in the Marine Red Alga Neopyropia yezoensis.

Heat stress responses are complex regulatory processes, including sensing, signal transduction, and gene expression. However, the exact mechanisms of these processes in seaweeds are not well known. We explored the relationship between membrane physical states and gene expression in the red alga Neopyropia yezoensis. To analyze heat-stress-induced gene expression, we identified two homologs of the heat-inducible high temperature response 2 (HTR2) gene in Neopyropia seriata, named NyHTR2 and NyHTR2L. We found conservation of HTR2 homologs only within the order Bangiales; their products contained a novel conserved cysteine repeat which we designated the Bangiales cysteine-rich motif. A quantitative mRNA analysis showed that expression of NyHTR2 and NyHTR2L was induced by heat stress. However, the membrane fluidizer benzyl alcohol (BA) did not induce expression of these genes, indicating that the effect of heat was not due to membrane fluidization. In contrast, expression of genes encoding multiprotein-bridging factor 1 (NyMBF1) and HSP70s (NyHSP70-1 and NyHSP70-2) was induced by heat stress and by BA, indicating that it involved a membrane-fluidization-dependent pathway. In addition, dark treatment under heat stress promoted expression of NyHTR2, NyHTR2L, NyMBF1, and NyHSP70-2, but not NyHSP70-1; expression of NyHTR2 and NyHTR2L was membrane-fluidization-independent, and that of other genes was membrane-fluidization-dependent. These findings indicate that the heat stress response in N. yezoensis involves membrane-fluidization-dependent and -independent pathways.

Research on the Regulatory Mechanism of Algae Reproduction under Abiotic Stress Conditions.

The intertidal and subtidal zones are characterized by daily and seasonal fluctuations in environmental conditions [...].

Heat-Stress Responses Differ among Species from Different 'Bangia' Clades of Bangiales (Rhodophyta).

The red alga 'Bangia' sp. ESS1, a 'Bangia' 2 clade member, responds to heat stress via accelerated asexual reproduction and acquires thermotolerance based on heat-stress memory. However, whether these strategies are specific to 'Bangia' 2, especially 'Bangia' sp. ESS1, or whether they are employed by all 'Bangia' species is currently unknown. Here, we examined the heat-stress responses of 'Bangia' sp. ESS2, a newly identified 'Bangia' clade 3 member, and Bangia atropurpurea. Intrinsic thermotolerance differed among species: Whereas 'Bangia' sp. ESS1 survived at 30 °C for 7 days, 'Bangia' sp. ESS2 and B. atropurpurea did not, with B. atropurpurea showing the highest heat sensitivity. Under sublethal heat stress, the release of asexual spores was highly repressed in 'Bangia' sp. ESS2 and completely repressed in B. atropurpurea, whereas it was enhanced in 'Bangia' sp. ESS1. 'Bangia' sp. ESS2 failed to acquire heat-stress tolerance under sublethal heat-stress conditions, whereas the acquisition of heat tolerance by priming with sublethal high temperatures was observed in both B. atropurpurea and 'Bangia' sp. ESS1. Finally, unlike 'Bangia' sp. ESS1, neither 'Bangia' sp. ESS2 nor B. atropurpurea acquired heat-stress memory. These findings provide insights into the diverse heat-stress response strategies among species from different clades of 'Bangia'.

The Absence of Hydrodynamic Stress Promotes Acquisition of Freezing Tolerance and Freeze-Dependent Asexual Reproduction in the Red Alga 'Bangia' sp. ESS1.

The ebb tide causes calm stress to intertidal seaweeds in tide pools; however, little is known about their physiological responses to loss of water movement. This study investigated the effects of static culture of 'Bangia' sp. ESS1 at 15 °C on tolerance to temperature fluctuation. The freezing of aer-obically cultured thalli at -80 °C for 10 min resulted in the death of most cells. By contrast, statically cultured thalli acquired freezing tolerance that increased cell viability after freeze-thaw cycles, although they did not achieve thermotolerance that would enable survival at the lethal temperature of 32 °C. Consistently, the unsaturation of membrane fatty acids occurred in static culture. Notably, static culture of thalli enhanced the release of asexual spores after freeze-and-thaw treatment. We conclude that calm stress triggers both the acquisition of freezing tolerance and the promotion of freezing-dependent asexual reproduction. These findings provide novel insights into stress toler-ance and the regulation of asexual reproduction in Bangiales.

Selection of AVP-Shortage Patients as Candidates for Low-Dose Oral Desmopressin Administration.

OBJECTIVE: We herein attempted to select male patients with an elevated nocturnal urinary frequency possibly due to a shortage of AVP. These patients may be good candidates for low-dose oral desmopressin administration. PATIENTS AND METHODS: Serum and spot urine osmolality, electrolytes, serum creatinine, casual blood glucose, plasma brain natriuretic polypeptide (BNP), and plasma AVP were measured at the same time in 97 elderly male patients with urinary symptoms under free water drinking. RESULTS: A binary plot of plasma AVP and serum osmolality indicated a region at which patients had relatively lower AVP considering higher serum osmolality. It was tentatively named the desmopressin region. Twenty out of 97 (20.6%) patients were in the desmopressin region. Daily urine output did not exceed 3 L in any patient. Urine osmolality was slightly lower in patients in the desmopressin region. No significant differences were observed in urine volume, urinary frequency, or urination questionnaire scores between both groups. CONCLUSION: AVP-shortage patients may be selected for treatment with oral desmopressin based on measurements of serum osmolality and plasma AVP.

Delayed Intracystic Hemorrhage after Percutaneous Drainage and Sclerotherapy for a Symptomatic Giant Hepatic Cyst: A Case Report.

Herein, we have reported a rare case of intracystic hemorrhage due to rupture of a right hepatic artery pseudoaneurysm in a 76-year-old female patient who underwent drainage and 3% polidocanol sclerotherapy for a symptomatic giant hepatic cyst. One month after sclerotherapy, the patient presented to the emergency room with acute and severe abdominal pain. Non-contrast T1-weighted magnetic resonance imaging findings showed high hepatic cyst fluid signal intensity and abdominal arteriography findings revealed a right hepatic artery pseudoaneurysm surrounding the hepatic cystic wall. Therefore, the patient was diagnosed with intracystic hemorrhage due to a ruptured pseudoaneurysm. Embolization, using a detachable coil, was successful. Interventional radiologists should be aware of potential vascular injuries during drainage and sclerotherapy for giant hepatic cysts.

Establishment of a Live-Imaging Analysis for Polarized Growth of Conchocelis in the Multicellular Red Alga Neopyropia yezoensis.

A wide range of tip-growing cells in plants display polarized cell growth, which is an essential cellular process for the form and function of individual cells. Understanding of the regulatory mechanisms underlying tip growth in terrestrial plants has improved. Cellular processes involved in tip growth have also been investigated in some algae species that form filamentous cells, but their regulatory mechanisms remain unclear. In the macro red alga Neopyropia yezoensis, for which genome information has recently been released, the conchocelis apical cell exhibits tip growth and forms a filamentous structure. Here, we report a live-imaging technique using high-resolution microscopy to analyze the tip growth and cell division of N. yezoensis conchocelis. This imaging analysis addressed tip growth dynamics and cell division in conchocelis apical cells. The directionality and tip growth expansion were disrupted by the application of cytoskeletal drugs, suggesting the involvement of microtubules (MTs) and actin filaments (AFs) in these processes. A growing apical cell mostly contained a single chloroplast that moved toward the expanding part of the apical cell. Drug application also inhibited chloroplast movement, implying that the movement may be dependent on the cytoskeleton. The study determined that live-imaging analysis is a versatile approach for exploring the dynamics of tip growth and cell division in N. yezoensis conchocelis, which provides insights into the regulatory mechanisms underlying cellular growth in multicellular red algae.

Low temperature causes discoloration by repressing growth and nitrogen transporter gene expression in the edible red alga Pyropia yezoensis.

The reduced availability of nitrogen sources in seawater leads to discoloration of the edible red seaweed Pyropia yezoensis and induces the expression of genes encoding ammonium, nitrate and urea transporters. In the present study, we demonstrate that low temperatures can also cause discoloration of this economically important seaweed. Thus, we addressed regulatory mechanisms of cold-inducible discoloration. When P. yezoensis thalli were incubated at 0, 5 and 10°C, the thalli exhibited retarded growth and discoloration, along with reduced phycoerythrin contents. Fertilization with nitrogen sources did not recover this discoloration at 0°C, suggesting that defects in nitrogen absorption cause low temperature-induced discoloration. The expression levels of many nitrogen transporter genes were reduced at low temperature. We propose that low temperature-mediated repression of ammonium, nitrate and urea transporter gene expression promotes the reduced absorption of nitrogen sources in P. yezoensis, thereby leading to discoloration. This process is different from the well-known mechanism underlying discoloration in P. yezoensis under nitrogen-deficient conditions at normal culture temperature.

Tumor-associated macrophage-derived inflammatory cytokine enhances malignant potential of malignant pleural mesothelioma.

Malignant pleural mesothelioma (MPM) is an asbestos-related aggressive malignant neoplasm. Due to the difficulty of achieving curative surgical resection in most patients with MPM, a combination chemotherapy of cisplatin and pemetrexed has been the only approved regimen proven to improve the prognosis of MPM. However, the median overall survival time is at most 12 mo even with this regimen. There has been therefore a pressing need to develop a novel chemotherapeutic strategy to bring about a better outcome for MPM. We found that expression of interleukin-1 receptor (IL-1R) was upregulated in MPM cells compared with normal mesothelial cells. We also investigated the biological significance of the interaction between pro-inflammatory cytokine IL-1ß and the IL-1R in MPM cells. Stimulation by IL-1ß promoted MPM cells to form spheroids along with upregulating a cancer stem cell marker CD26. We also identified tumor-associated macrophages (TAMs) as the major source of IL-1ß in the MPM microenvironment. Both high mobility group box 1 derived from MPM cells and the asbestos-activated inflammasome in TAMs induced the production of IL-1ß, which resulted in enhancement of the malignant potential of MPM. We further performed immunohistochemical analysis using clinical MPM samples obtained from patients who were treated with the combination of platinum plus pemetrexed, and found that the overexpression of IL-1R tended to correlate with poor overall survival. In conclusion, the interaction between MPM cells and TAMs through a IL-1ß/IL-1R signal could be a promising candidate as the target for novel treatment of MPM (Hyogo College of Medicine clinical trial registration number: 2973).

Irregularly branched microvessels as visualized by magnifying endoscopy: a reliable marker for predicting deep submucosal invasion of superficial esophageal squamous cell carcinoma.

Background and study aims Magnifying endoscopy with narrow-band imaging (M-NBI) is reported to be useful in diagnosing invasion depth of superficial esophageal squamous cell carcinoma (SCC), but accurate diagnosis of deep submucosal invasion (SM2) has remained difficult. However, we discovered that irregularly branched microvessels observed with M-NBI are detected in SM2 cancers with high prevalence. Thus, this retrospective study aimed to investigate the diagnostic performance of irregularly branched microvessels as visualized by M-NBI for predicting SM2 cancers. Patients and methods Patients with superficial esophageal SCC lesions that were endoscopically or surgically resected at our hospital between September 2005 and December 2014 were included. Endoscopic findings by M-NBI of these lesions were presented to an experienced endoscopist who was unaware of the histopathological diagnosis and who then judged whether irregularly branched microvessels were present. Using the invasion depth according to postoperative histopathological diagnosis as the gold standard, we determined the diagnostic performance of the presence of irregularly branched microvessels as an indicator for SM2 cancers. Results A total of 302 superficial esophageal SCC lesions (228 patients) were included in the analysis. When irregularly branched microvessels were used as an indicator of SM2 cancers, the diagnostic accuracy was 94.0 % (95 % confidence interval [CI]: 91.1-96.1 %), sensitivity was 79.4 % (95 % CI: 66.6-88.4 %), specificity was 95.9 % (95 % CI: 94.3-97.0 %), positive predictive value was 71.1 % (95 % CI: 59.6-79.1 %), and negative predictive value was 97.3 % (95% CI: 95.7-98.5 %). Conclusions Irregularly branched microvessels may be a reliable M-NBI indicator for the diagnosis of cancers with deep submucosal invasion.