[Hypervascular Tumors such as Hemangioblastoma].

Hemangioblastomas are richly vascular tumors. Therefore, visualizing the vascular anatomy of their feeders and drainers is important for planning surgical excision. Preoperative three-dimensional computer graphic(3DCG)images are useful for determining the number, location, and course of their feeders and drainers.

Links between cholesteryl sulfate-dependent and -independent processes in the morphological and physiological changes of Entamoeba encystation.

The protozoan parasite Entamoeba histolytica causes amoebiasis, a global public health problem. Amoebiasis is solely transmitted by cysts that are produced from proliferative trophozoites by encystation in the large intestine of humans. During encystation, various metabolites, pathways, and cascades sequentially orchestrate the morphological and physiological changes required to produce cysts. Cholesteryl sulfate (CS) has recently been revealed to be among the key molecules that control the morphological and physiological changes of encystation by exerting pleiotropic effects. CS promotes the rounding of encysting Entamoeba cells and maintains this spherical morphology as encysting cells are surrounded by the cyst wall, a prerequisite for resistance against environmental stresses. CS is also involved in the development of membrane impermeability, another prerequisite for resistance. The initiation of cyst wall formation is, however, CS-independent. Here, we overview CS-dependent and -independent processes during encystation and discuss their functional linkage. We also discuss a potential transcriptional cascade that controls the processes necessary to produce dormant Entamoeba cysts.

CD47 promotes peripheral T cell survival by preventing dendritic cell-mediated T cell necroptosis.

Conventional dendritic cells (cDCs) are required for peripheral T cell homeostasis in lymphoid organs, but the molecular mechanism underlying this requirement has remained unclear. We here show that T cell-specific CD47-deficient (Cd47 ΔT) mice have a markedly reduced number of T cells in peripheral tissues. Direct interaction of CD47-deficient T cells with cDCs resulted in activation of the latter cells, which in turn induced necroptosis of the former cells. The deficiency and cell death of T cells in Cd47 ΔT mice required expression of its receptor signal regulatory protein α on cDCs. The development of CD4+ T helper cell-dependent contact hypersensitivity and inhibition of tumor growth by cytotoxic CD8+ T cells were both markedly impaired in Cd47 ΔT mice. CD47 on T cells thus likely prevents their necroptotic cell death initiated by cDCs and thereby promotes T cell survival and function.

Possible Effect of Blonanserin Transdermal Patch on Antiemetic Control in Patients With Terminal Cancer with Refractory Nausea.

Background: Haloperidol is widely used for antiemetic control in advanced cancer. However, due to its limited administration methods (oral or injection), its management is frequently challenging in palliative home care. Recently, a blonanserin transdermal patch was developed as the first antipsychotic percutaneous agent. Objectives: This study aimed to evaluate its effectiveness and safety for refractory nausea. Methods: We conducted an observational study of 21 terminal cancer patients who had been initiated for refractory nausea in their homes. Results: After its initiation, none of the patients experienced aggravated nausea, and the number of patients with severe nausea decreased dramatically (52.4% vs. 9.5%, p = 0.008). Of 16 patients without ascites, 12.5% had not improved their nausea, which was lower than in patients with ascites (80.0%). Conclusions: Blonanserin transdermal patch has a possible effect on antiemetic control in cancer patients, and its efficacy might be particularly prominent in patients without ascites.

Efficacy of blonanserin transdermal patch on terminal delirium in patients with respiratory diseases.

BACKGROUND: Delirium is a common distressing symptom observed in patients with terminal respiratory diseases and is treated with antipsychotic medications such as haloperidol. Its management is difficult, especially in palliative home care, because its administration is limited to oral or injection methods. Recently, the blonanserin transdermal patch was developed as the first antipsychotic percutaneous agent. After it became available, we recognized its potential for the management of delirium and the alleviation of uncontrolled dyspnea in clinical practice. Thus, this study aimed to assess its efficacy in patients with terminal respiratory diseases. METHODS: This retrospective study included 113 patients with respiratory diseases who were cared for at home. The efficacy was evaluated through the prevalence of terminal delirium before and after its treatment initiation for uncontrolled dyspnea. RESULTS: Blonanserin transdermal patch treatment for uncontrolled dyspnea improved the prevalence and severity of terminal delirium (from 70.4% to 16.3%, p < 0.001) and reduced the number of doctors' visits to patients' homes within a week before their death (from 4.0 to 3.0, p = 0.086). A sub-group analysis of 23 patients with interstitial pneumonia revealed that the treatment prevented dyspnea progression by inhibiting terminal delirium. CONCLUSIONS: Blonanserin transdermal patch performed similarly to haloperidol, as previously reported, for managing terminal delirium. Our study suggests that a blonanserin transdermal patch potentially prevents terminal delirium and alleviates uncontrolled dyspnea in patients with respiratory diseases. Our findings encourage clinical trials to evaluate the safety and efficacy of blonanserin transdermal patches for patients with terminal illnesses.

Thermal Effect on Thin-Film Formation of the Polymer Sheets by the CO2 Laser with the Copper Base.

A method that makes polymer sheets partially thinner with continuous-wave carbon dioxide (CO2) lasers has been developed. This method can create thin polymer films by attaching the polymer sheets to the copper base by vacuum suction through the holes in the base. Applying the method to polypropylene (PP), polyethylene terephthalate (PET), polystyrene (PS), and polytetrafluoroethylene (PTFE), the thin-film formation is confirmed in PP, PET, and PS but not PTFE. These polymers have the similar thermal properties. PP, PET, and PS show fluidity with increased temperature, but PTFE does not have fluidity. These characteristics of the polymers indicate that the fluidity of polymer is the important characteristic for film formation. The experiments with PP and PET sheets of different thickness show that thicker sheets make thicker films. The fluid flow of the molten polymer is considered to form the thin film at the bottom of the groove made by laser scribing. The numerical simulation of the 2D thermal model also indicates the week cooling effects of the base on the film formation and importance of polymer fluidity. The results of Fourier transform infrared spectrometer (FT-IR) show thermal degradation of the films. To decrease the heat's effect on the films, the polymer sheets should be processed at the highest laser-beam scanning speed that can make thin films.

Pleiotropic Roles of Cholesteryl Sulfate during Entamoeba Encystation: Involvement in Cell Rounding and Development of Membrane Impermeability.

Entamoeba histolytica, a protozoan parasite, causes amoebiasis, which is a global public health problem. The major route of infection is oral ingestion of cysts, the only form that is able to transmit to a new host. Cysts are produced by cell differentiation from proliferative trophozoites in a process termed "encystation." During encystation, cell morphology is markedly changed; motile amoeboid cells become rounded, nonmotile cells. Concomitantly, cell components change and significant fluctuations of metabolites occur. Cholesteryl sulfate (CS) is a crucial metabolite for encystation. However, its precise role remains uncertain. To address this issue, we used in vitro culture of Entamoeba invadens as the model system for the E. histolytica encystation study and identified serum-free culture conditions with CS supplementation at concentrations similar to intracellular CS concentrations during natural encystation. Using this culture system, we show that CS exerts pleiotropic effects during Entamoeba encystation, affecting cell rounding and development of membrane impermeability. CS dose dependently induced and maintained encysting cells as spherical maturing cysts with almost no phagocytosis activity. Consequently, the percentage of mature cysts was increased. CS treatment also caused time- and dose-dependent development of membrane impermeability in encysting cells via induction of de novo synthesis of dihydroceramides containing very long N-acyl chains (≥26 carbons). These results indicate that CS-mediated morphological and physiological changes are necessary for the formation of mature cysts and the maintenance of the Entamoeba life cycle. Our findings also reveal important morphological aspects of the process of dormancy and the control of membrane structure. IMPORTANCE Entamoeba histolytica causes a parasitic infectious disease, amoebiasis. Amoebiasis is a global public health problem with a high occurrence of infection and inadequate clinical options. The parasite alternates its form between a proliferative trophozoite and a dormant cyst that enables the parasite to adapt to new environments. The transition stage in which trophozoites differentiate into cysts is termed "encystation." Cholesteryl sulfate is essential for encystation; however, its precise role remains to be determined. Here, we show that cholesteryl sulfate is a multifunctional metabolite exerting pleiotropic roles during Entamoeba encystation, including the rounding of cells and the development of membrane impermeability. Such morphological and physiological changes are required for Entamoeba to produce cysts that are transmissible to a new host, which is essential for maintenance of the Entamoeba life cycle. Our findings are therefore relevant not only to Entamoeba biology but also to general cell and lipid biology.

A growing aneurysm of the posterior inferior cerebellar artery complicated with cerebellar infarction: A case report.

INTRODUCTION AND IMPORTANCE: Hereby we describe an instructive patient with cerebellar infarction and a growing aneurysm at the posterior inferior cerebellar artery (PICA), which was not a true cause of infarction. CASE PRESENTATION: A 50-year-old female presented with dizziness and posterior neck pain at our hospital (Mitaka city, Tokyo, Japan). Diffusion weighted magnetic resonance (MR) images showed cerebellar infarction in the left PICA territory and MR angiography study showed an aneurysm at the origin of the left PICA, which grew in 2 weeks. Since we considered cerebellar infarction was caused by thrombosis from the aneurysm, trapping of the PICA and occipital artery-PICA bypass was performed to prevent recurrent cerebellar infarction and rupture of the aneurysm by neurosurgeons. During the operation, dissection was observed at the distal PICA, which was diagnosed to be the true cause of cerebellar infarction. By the follow-up for 12 months at an outpatient, there was no recurrence of cerebral infarction. CLINICAL DISCUSSION: A specimen of the artery showing the findings of dissection was not obtained, and the pathological diagnosis could not be made. It would be controversial whether a surgical procedure presented here was the most optimal. CONCLUSION: This is a first reported case of growing aneurysms and cerebral infarction due to arterial dissection. Even if cerebral infarction is accompanied by growing aneurysms, arterial dissection should be included in the differential diagnoses of a cause of infarction. Posterior cervical pain can be a clue for early appropriate diagnosis in such a case.

Entamoeba Chitinase is Required for Mature Round Cyst Formation.

Entamoeba histolytica, a protozoan parasite, causes amoebiasis in humans. Amoebiasis transmission is solely mediated by chitin-walled cysts, which are produced in the large intestine of humans from proliferative trophozoites by a cell differentiation process called encystation. Resistance to environmental stresses, an essential characteristic for transmission, is attributed to the cyst wall, which is constructed from chitin and several protein components, including chitinase. Chitinase may play a key role in cyst wall formation; however, this has not been confirmed. Here, to elucidate the physiological role of chitinase during Entamoeba encystation, we identified a new chitinase inhibitor, 2,6-dichloro-4-[2-(1-piperazinyl)-4-pyridinyl]-N-(1,3,5-trimethyl-1H-pyrazol-4-yl)-benzenesulfonamide, by recombinant-Entamoeba chitinase-based screening of 400 Pathogen Box chemicals. This compound dose dependently inhibited native chitinase associated with Entamoeba invadens encystation, a model for E. histolytica encystation, with an 50% inhibitory concentration (IC50) of ∼0.6 µM, which is comparable to the IC50s (0.2 to 2.5 µM) for recombinant E. histolytica and E. invadens chitinases. Furthermore, the addition of this compound to E. invadens encystation-inducing cultures increased the generation of cyst walls with an abnormal shape, the most characteristic of which was a "pot-like structure." A similar structure also appeared in standard culture, but at a far lower frequency. These results indicate that chitinase inhibition increases the number of abnormal encysting cells, thereby significantly reducing the efficiency of cyst formation. Transmission electron microscopy showed that compound-treated encysting cells formed an abnormally loose cyst wall and an unusual gap between the cyst wall and cell membrane. Hence, Entamoeba chitinase is required for the formation of mature round cysts. IMPORTANCE Amoebiasis is caused by Entamoeba histolytica infection and is transmitted by dormant Entamoeba cells or cysts. Cysts need to be tolerant to severe environmental stresses faced outside and inside a human host. To confer this resistance, Entamoeba parasites synthesize a wall structure around the cell during cyst formation. This cyst wall consists of chitin and several protein components, including chitinase. The physiological roles of these components are not fully understood. Here, to elucidate the role of chitinase during cyst formation, we identified a new chitinase inhibitor by screening a library of 400 compounds. Using this inhibitor, we showed that chitinase inhibition causes the formation of abnormal cyst walls, the most characteristic of which is a "pot-like structure." This results in decreased production of mature cysts. Chitinase is therefore required for Entamoeba to produce mature cysts for transmission to a new host.

High Carbohydrate Diet Increased Glucose Transporter Protein Levels in Jejunum but Did Not Lead to Enhanced Post-Exercise Skeletal Muscle Glycogen Recovery.

We examined the effect of dietary carbohydrate intake on post-exercise glycogen recovery. Male Institute of Cancer Research (ICR) mice were fed moderate-carbohydrate chow (MCHO, 50%cal from carbohydrate) or high-carbohydrate chow (HCHO, 70%cal from carbohydrate) for 10 days. They then ran on a treadmill at 25 m/min for 60 min and administered an oral glucose solution (1.5 mg/g body weight). Compared to the MCHO group, the HCHO group showed significantly higher sodium-D-glucose co-transporter 1 protein levels in the brush border membrane fraction (p = 0.003) and the glucose transporter 2 level in the mucosa of jejunum (p = 0.004). At 30 min after the post-exercise glucose administration, the skeletal muscle and liver glycogen levels were not significantly different between the two diet groups. The blood glucose concentration from the portal vein (which is the entry site of nutrients from the gastrointestinal tract) was not significantly different between the groups at 15 min after the post-exercise glucose administration. There was no difference in the total or phosphorylated states of proteins related to glucose uptake and glycogen synthesis in skeletal muscle. Although the high-carbohydrate diet significantly increased glucose transporters in the jejunum, this adaptation stimulated neither glycogen recovery nor glucose absorption after the ingestion of post-exercise glucose.

PKD1-Associated Arachnoid Cysts in Autosomal Dominant Polycystic Kidney Disease.

OBJECTIVES: the prevalence of intracranial aneurysms and arachnoid cysts is higher in patients with autosomal dominant polycystic kidney disease (ADPKD) than in the general population. A genotype correlation was reported for intracranial aneurysms, but it is unclear for arachnoid cysts. Therefore, the genotype correlation with intracranial aneurysms and arachnoid cysts was investigated in ADPKD. MATERIALS AND METHODS: intracranial aneurysms and arachnoid cysts were screened by magnetic resonance imaging (MRI), and PKD genotypes were examined using next-generation sequencing for 169 patients with ADPKD. RESULTS: PKD1-, PKD2- and no-mutation were identified in 137, 24 and 8 patients, respectively. Intracranial aneurysms and arachnoid cysts were found in 34 and 25 patients, respectively, with no significant difference in frequency. Genotype, sex, estimated glomerular filtration rate and age at ADPKD diagnosis significantly affected the age at brain MRI. The proportional hazard risk analyzed using the age at brain MRI adjusted by these four variables was 5.0-times higher in the PKD1 group than in the PKD2 group for arachnoid cysts (P = 0.0357), but it was not different for intracranial aneurysms (P = 0.1605). Arachnoid cysts were diagnosed earlier in the PKD1 group than in the PKD2 group (54.8 vs 67.7 years, P = 0.0231), but no difference was found for intracranial aneurysms (P = 0.4738) by Kaplan-Meier analysis. CONCLUSIONS: this study demonstrated the correlation between arachnoid cysts and PKD1 mutation. The reported association of arachnoid cysts with advanced renal disease may be due to the common correlation of these factors with PKD1 mutation.

Thin-Film Processing of Polypropylene and Polystyrene Sheets by a Continuous Wave CO2 Laser with the Cu Cooling Base.

Carbon dioxide (CO2) laser is widely used in commercial and industrial fields to process various materials including polymers, most of which have high absorptivity in infrared spectrum. Thin-film processing by the continuous wave (CW) laser is difficult since polymers are deformed and damaged by the residual heat. We developed the new method to make polypropylene (PP) and polystyrene (PS) sheets thin. The sheets are pressed to a Cu base by extracting air between the sheets and the base during laser processing. It realizes to cut the sheets to around 50 µm thick with less heat effects on the backside which are inevitable for thermal processing using the CW laser. It is considered that the boundary between the sheets and the base is in thermal equilibrium and the base prevents the sheets from deforming to support the backside. The method is applicable to practical use since it does not need any complex controls and is easy to install to an existing equipment with a minor change of the stage.

TREM2 is a receptor for non-glycosylated mycolic acids of mycobacteria that limits anti-mycobacterial macrophage activation.

Mycobacterial cell-wall glycolipids elicit an anti-mycobacterial immune response via FcRγ-associated C-type lectin receptors, including Mincle, and caspase-recruitment domain family member 9 (CARD9). Additionally, mycobacteria harbor immuno-evasive cell-wall lipids associated with virulence and latency; however, a mechanism of action is unclear. Here, we show that the DAP12-associated triggering receptor expressed on myeloid cells 2 (TREM2) recognizes mycobacterial cell-wall mycolic acid (MA)-containing lipids and suggest a mechanism by which mycobacteria control host immunity via TREM2. Macrophages respond to glycosylated MA-containing lipids in a Mincle/FcRγ/CARD9-dependent manner to produce inflammatory cytokines and recruit inducible nitric oxide synthase (iNOS)-positive mycobactericidal macrophages. Conversely, macrophages respond to non-glycosylated MAs in a TREM2/DAP12-dependent but CARD9-independent manner to recruit iNOS-negative mycobacterium-permissive macrophages. Furthermore, TREM2 deletion enhances Mincle-induced macrophage activation in vitro and inflammation in vivo and accelerates the elimination of mycobacterial infection, suggesting that TREM2-DAP12 signaling counteracts Mincle-FcRγ-CARD9-mediated anti-mycobacterial immunity. Mycobacteria, therefore, harness TREM2 for immune evasion.

Glucosyl Hesperidin Has an Anti-diabetic Effect in High-Fat Diet-Induced Obese Mice.

Glucosyl hesperidin (GH) is a water-soluble derivative of hesperidin, a citrus flavonoid. GH has various pharmacological effects, such as hypolipidemic and hypouricemic effects, and may therefore be a useful supplement or drug. In the present study, we evaluated the effects of long- and short-term intake of GH on hyperglycemia and macrophage infiltration into the adipose tissue of high-fat diet (HFD)-fed mice. Long-term (11-week) consumption of GH tended to reduce body weight and the fasting blood glucose concentration of the HFD-fed mice, and ameliorated glucose intolerance and insulin resistance, according to glucose and insulin tolerance tests. Additionally, although GH did not affect fat pad weight, it reduced HFD-induced macrophage infiltration into adipose tissue. Short-term (2-week) consumption of GH did not affect the HFD-induced increases in body weight or fasting blood glucose, and it did not ameliorate glucose intolerance or insulin resistance. However, short-term intake did reduce the HFD-induced macrophage infiltration and monocyte chemotactic protein 1 (MCP-1) expression in adipose tissue. Furthermore, hesperetin, which is an aglycone of GH, inhibited MCP-1 expression in 3T3-L1 adipocytes, 3T3-L1 adipocytes co-cultured with RAW264 macrophages, and tumor necrosis factor-α-treated 3T3-L1 adipocytes. The present findings suggest that daily consumption of GH may have preventive and/or therapeutic effects on obesity-related diseases, such as diabetes mellitus.

Stage-Specific De Novo Synthesis of Very-Long-Chain Dihydroceramides Confers Dormancy to Entamoeba Parasites.

Amoebiasis is a parasitic disease caused by Entamoeba histolytica infection and is a serious public health problem worldwide due to ill-prepared preventive measures as well as its high morbidity and mortality rates. Amoebiasis transmission is solely mediated by cysts. Cysts are produced by the differentiation of proliferative trophozoites in a process termed "encystation." Entamoeba encystation is a fundamental cell differentiation process and proceeds with substantial changes in cell metabolites, components, and morphology, which occur sequentially in an orchestrated manner. Lipids are plausibly among these metabolites that function as key factors for encystation. However, a comprehensive lipid analysis has not been reported, and the involved lipid metabolic pathways remain largely unknown. Here, we exploited the state-of-the-art untargeted lipidomics and characterized 339 molecules of 17 lipid subclasses. Of these, dihydroceramide (Cer-NDS) was found to be among the most induced lipid species during encystation. Notably, in encysting cells, amounts of Cer-NDS containing very long N-acyl chains (≥26 carbon) were more than 30-fold induced as the terminal product of a de novo metabolic pathway. We also identified three ceramide synthase genes responsible for producing the very-long-chain Cer-NDS molecules. These genes were upregulated during encystation. Furthermore, these ceramide species were shown to be indispensable for generating membrane impermeability, a prerequisite for becoming dormant cyst that shows resistance to environmental assault inside and outside the host for transmission. Hence, the lipid subclass of Cer-NDS plays a crucial role for Entamoeba cell differentiation and morphogenesis by alternating the membrane properties.IMPORTANCEEntamoeba is a protozoan parasite that thrives in its niche by alternating its two forms between a proliferative trophozoite and dormant cyst. Cysts are the only form able to transmit to a new host and are differentiated from trophozoites in a process termed "encystation." During Entamoeba encystation, cell metabolites, components, and morphology drastically change, which occur sequentially in an orchestrated manner. Lipids are plausibly among these metabolites. However, the involved lipid species and their metabolic pathways remain largely unknown. Here, we identified dihydroceramides (Cer-NDSs) containing very long N-acyl chains (C26 to C30) as a key metabolite for Entamoeba encystation by our state-of-the-art untargeted lipidomics. We also showed that these Cer-NDSs are critical to generate the membrane impermeability, a prerequisite for this parasite to show dormancy as a cyst that repels substances and prevents water loss. Hence, ceramide metabolism is essential for Entamoeba to maintain the parasitic lifestyle.

Effects of 5-mg dose of olanzapine for breakthrough nausea and vomiting in patients receiving carboplatin-based chemotherapy: a prospective trial.

BACKGROUND: Olanzapine 10 mg is recommended for breakthrough chemotherapy-induced nausea and vomiting. However, there is a possibility that 5 mg can be expected to be sufficiently effective. We aimed to investigate the efficacy and safety of olanzapine 5 mg for breakthrough chemotherapy-induced nausea and vomiting. METHODS: A single-arm prospective trial of olanzapine 5 mg every 24 h for 72 h was conducted to treat breakthrough chemotherapy-induced nausea and vomiting in patients receiving carboplatinbased chemotherapy. The primary endpoint was total control (i.e., no emesis, no nausea, and no rescue medications) over 72 h. The secondary endpoints were early efficacy using the nausea scores at 30, 60, and 120 min after taking olanzapine from baseline and adverse events. RESULTS: Among 84 potentially eligible patients, 19 patients who took olanzapine for breakthrough chemotherapy-induced nausea and vomiting were examined. The total control rate was 32% (95% CI: 13- 57%), 65% (95% CI: 38-89%), 65% (95% CI: 38-89%), and 29% (95% CI: 10-56%) during 2-24, 24-48, 48-72 h, and overall period, respectively. The nausea scale significantly reduced after 30 min (P=0.0078), and the scale had been reduced by 67% from the baseline after 60 min. The adverse event of somnolence of any grade was observed in 13 (68%) patients, 6 (32%) of whom had grade 2 and 1 (5%) grade 3 somnolence. CONCLUSIONS: Olanzapine 5 mg did not show the expected effect on the complete disappearance of breakthrough chemotherapy-induced nausea and vomiting within 24 h.

Discovery of Self-Assembling Small Molecules as Vaccine Adjuvants.

Immune potentiators, termed adjuvants, trigger early innate immune responses to ensure the generation of robust and long-lasting adaptive immune responses of vaccines. Presented here is a study that takes advantage of a self-assembling small-molecule library for the development of a novel vaccine adjuvant. Cell-based screening of the library and subsequent structural optimization led to the discovery of a simple, chemically tractable deoxycholate derivative (molecule 6, also named cholicamide) whose well-defined nanoassembly potently elicits innate immune responses in macrophages and dendritic cells. Functional and mechanistic analyses indicate that the virus-like assembly enters the cells and stimulates the innate immune response through Toll-like receptor 7 (TLR7), an endosomal TLR that detects single-stranded viral RNA. As an influenza vaccine adjuvant in mice, molecule 6 was as potent as Alum, a clinically used adjuvant. The studies described here pave the way for a new approach to discovering and designing self-assembling small-molecule adjuvants against pathogens, including emerging viruses.

The dynamics of ultrastructural changes during Entamoeba invadens encystation.

Entamoeba histolytica infection causes amoebiasis, which is a global public health problem. The major route of infection is oral ingestion of E. histolytica cysts, cysts being the sole form responsible for host-to-host transmission. Cysts are produced by cell differentiation from proliferative trophozoites in a process termed 'encystation'. Therefore, encystation is an important process from a medical as well as a biological perspective. Previous electron microscopy studies have shown the ultrastructure of precysts and mature cysts; however, the dynamics of ultrastructural changes during encystation were ambiguous. Here, we analysed a series of Entamoeba invadens encysting cells by transmission electron microscopy. Entamoeba invadens is a model for encystation and the cells were prepared by short interval time course sampling from in vitro encystation-inducing cultures. We related sampled cells to stage conversion, which was monitored in the overall population by flow cytometry. The present approach revealed the dynamics of ultrastructure changes during E. invadens encystation. Importantly, the results indicate a functional linkage of processes that are crucial in encystation, such as glycogen accumulation and cyst wall formation. Hence, this study provides a reference for studying sequential molecular events during Entamoeba encystation.

[Necessity of Serum Calcium Concentration Measurement for Investigating Hypocalcemia Induced by Denosumab].

Denosumab is widely used for treating bone metastases in patients with advanced cancer. However, hypocalcemia has been reported as a serious adverse effect during this treatment. Therefore, monitoring serum calcium concentration is essential. During long-term continuous administration ofdenosumab, the burden ofeach blood sampling occasion and medical economics should be clarified; however, the appropriate blood sampling frequency has not yet been confirmed. In the present study, we performed a retrospective investigation of serum calcium concentration after denosumab administration. The results indicated that serum calcium concentration tended to increase with repeated administration. A significant increase was observed at the time ofthe third and sixth administration. This suggested that it was possible to reduce the frequency ofblood sampling during long-term administration ofdenosumab with appropriate calcium supplementation.

Adjuvant activity of Mycobacteria-derived mycolic acids.

Successful vaccination, especially with safe vaccines such as component/subunit vaccines, requires proper activation of innate immunity and, for this purpose, adjuvant is used. For clinical use, alum is frequently used while, for experimental use, CFA, containing Mycobacterial components, was often used. In this report, we demonstrated that mycolic acids (MA), major and essential lipid components of the bacterial cell wall of the genus Mycobacterium, has adjuvant activity. MA plus model antigen-immunization induced sufficient humoral response, which was largely comparable to conventional CFA plus antigen-immunization. Importantly, while CFA plus antigen-immunization induced Th17-biased severe and destructive inflammatory responses at the injected site, MA plus antigen-immunization induced Th1-biased mild inflammation at the site. MA induced dendritic cell activation by co-stimulatory molecule induction as well as inflammatory cytokine/chemokine induction. MA plus antigen-immunization successfully protected mice from tumor progression both in prevention and in therapy models. We thus submit that MA is a promising adjuvant candidate material for clinical purposes and for experimental purposes from a perspective of animal welfare.