DCLK1 Expression and its Functional Significance in Intrahepatic Cholangiocarcinoma and Bil-IN Stage.

BACKGROUND/AIM: Although several studies in some neoplasms have reported correlation between the expression levels of Doublecortin-like kinase1(DCLK1) and carcinogenesis, its role in cholangiocarcinoma remains unknown. MATERIALS AND METHODS: DCLK1 expression in normal epithelium (NE), biliary intraepithelial neoplasia (BilIN)1∼3, and intrahepatic cholangiocarcinoma (ICC) were investigated immuno-histochemically. The molecular effects of DCLK1 were investigated by gene silencing using RNAi [DCLK1-tagrgeting (siDCLK1)]. The human ICC cell lines HuCCT1 and HuH28 were transfected with these siRNAs, and used for assays in the presence or absence of DCLK1 inhibitors. RESULTS: The positive ratio of DCLK1 expression in ICC was higher than that in NE, and equally distributed among BilIN1∼3 (NE: BilIN1: BilIN2: BilIN3: ICC=62%: 91%: 97%: 100%: 95%, p<0.05). In the wound healing assay, the migration of the siDCLK1-treated cells was significantly inhibited compared to the NT-treated cells (p<0.05). In the cell invasion assay, the invasion of the siDCLK1-treated cells was significantly inhibited compared to the NT-treated cells (p<0.05). In the presence of the DCLK1 inhibitor, cell proliferative capacity at 24 hours was decreased in a concentration-dependent manner. CONCLUSION: DCLK1 was highly expressed in the early stage of ICC carcinogenesis. Human ICC cell growth was suppressed in vitro by siRNA silencing of DCLK1 or after treatment with the DCLK1 inhibitor, indicating DCLK1 may be molecular target for ICC therapy.

Trajectories of medical care expenditure in the last year of life associated with long-term care utilization in frail older adults: A retrospective cohort study.

BACKGROUND: Medical care and long-term care utilization in the last year of life of frail older adults could be a key indicator of their quality of life. This study aimed to identify the medical care expenditure (MCE) trajectories in the last year of life of frail older adults by investigating the association between MCE and long-term care utilization in each trajectory. METHODS: The retrospective cohort study of three municipalities in Japan included 405 decedents (median age at death, 85 years; 189 women [46.7%]) from a cohort of 1,658 frail older adults aged ≥65 years who were newly certified as support level in the long-term care insurance program from April 2012 to March 2013. This study used long-term care and medical insurance claim data from April 2012 to March 2017. The primary outcome was MCE over the 12 months preceding death. Group-based trajectory modeling was conducted to identify the MCE trajectories. A mixed-effect model was employed to examine the association between long-term care utilization and MCE in each trajectory. RESULTS: Participants were stratified into four groups based on MCE trajectories over the 12 months preceding death as follows: rising (n = 159, 39.3%), persistently high (n = 143, 35.3%), minimal (n = 56, 13.8%), and descending (n = 47, 11.6%) groups. Home-based long-term care utilization was associated with increased MCE in the descending trajectory (coefficient, 1.48; 95% confidence interval [CI], 1.35-1.62). Facility-based long-term care utilization was associated with reduced MCE in the rising trajectory (coefficient, 0.59; 95% CI, 0.50-0.69). Both home-based (coefficient, 0.92; 95% CI, 0.85-0.99) and facility-based (coefficient; 0.53; 95% CI, 0.41-0.63) long-term care utilization were associated with reduced MCE in the persistently high trajectory. CONCLUSIONS: These findings may facilitate the integration of medical and long-term care models at the end of life in frail older adults.

Do Home- and Community-Based Services Delay Frailty Onset in Older Adults With Low Care Needs?

OBJECTIVES: To assess whether using adult day services or personal assistance services can delay the onset of frailty among older adults with low care needs during a 5-year follow-up study. DESIGN: This prospective cohort study was conducted using long-term care and health insurance claims data. SETTING AND PARTICIPANTS: This was a population-based study of 3 municipalities in Osaka, Japan. Initially, 655 nonfrail or prefrail individuals were included from a cohort of 790 population-based adults aged ≥65 years, who were newly certified as being on a support level of the long-term care insurance program from September 2012 to March 2013. METHODS: Using long-term care and health insurance claims data from the Southern Osaka Health and Aging Study, conducted between April 2012 and March 2017, monthly usage of adult day and personal assistance services was measured. Data were analyzed from December 2021 to January 2022. RESULTS: Of the 655 individuals (median age at baseline: 79 years), 436 (66.6%) were female, 388 (59.2%) were nonfrail, and 267 (40.8%) were prefrail, according to the Veterans Affairs Frailty Index. During the 5-year follow-up period, 222 individuals (33.9%) experienced the onset of frailty. The time-dependent Cox regression models showed that using adult day services lowered the risk of frailty when compared with not using such services [hazard ratio (HR) 0.60, 95% CI 0.42-0.86; P = .006], although personal assistance services usage was not associated with the onset of frailty (HR 0.70, 95% CI 0.48-1.03, P = .07). CONCLUSIONS AND IMPLICATIONS: Using adult day services lowered the risk of frailty in older adults with low care needs over the 5-year follow-up period. The findings support the value of providing adult day services to prevent frailty for those in need of long-term care.

Five-year disease-related risk of mortality in ambulatory frail older Japanese.

Objectives　We investigated the 5-year disease-related mortality risk, including that associated with neoplasms, mental/behavioral/neurodevelopmental disorders, and diseases of the circulatory system and respiratory system,in ambulatory frail Japanese older adults.Methods　We retrospectively analyzed long-term care and health insurance claims data in this cohort study performed between April 2012 and March 2017. The primary outcome was mortality, and the secondary outcome was care-need level decline. Risk factors were determined based on the International Statistical Classification of Disease and Related Health Problems, 10th Revision codes, hospitalization, and institutionalization. The study included 1,239 ambulatory frail older adults newly certified as needing Support-Level care at baseline (April 2012-March 2013) across three Japanese municipalities.Results　Of the 1,239 participants, 454 (36.6%) died. Neoplasms (hazard ratio [HR] 2.69, 95% confidence interval [CI] 1.97-3.68) or respiratory system diseases (HR 1.62, 95%CI 1.26-2.08) were independently associated with mortality. Mental/behavioral/neurodevelopmental disorders (HR 1.39, 95%CI 1.17-1.66) or diseases of the respiratory system(HR 86, 95%CI 75-99) were independently associated with care-need level decline.Conclusions　This study suggests that neoplasms or respiratory system diseases were associated with a high mortality risk and that mental/behavioral/neurodevelopmental disorders were associated with care-need level decline among ambulatory frail older adults. Optimal disease management and effective long-term care are important to delay the onset of these events in older adults certified as needing Support-Level care.

[A Case of Neuromyelitis Optica Spectrum Disease with Hypoglycorrhachia].

A 75-year-old Japanese woman developed myelitis 3years prior to her admission. She was diagnosed with HTLV-1-related myelitis and had taken prednisolone. Her myelitis relapsed several times, and serum aquaporin-4 was positive in an ELISA. She developed a sudden headache, consciousness disturbance, dysarthria, and left limb paralysis, and was admitted to our hospital. The CSF analysis revealed pleocytosis dominated by morphonuclear cells and hypoglycorrhachia. Magnetic resonance imaging revealed abnormalities in the corpus callosum, bilateral thalamus, and corticospinal tracts. We initially suspected a relapse of neuromyelitis optica spectrum disorder (NMOSD) and infection. We treated the patient with methylprednisolone pulse and antibacterial and antiviral treatment, which were not effective. Plasmapheresis was performed five times, and she gradually improved. Immunosuppressive treatment was added. It is rare for NMOSD to cause hypoglycorrhachia. This case suggests that infection may trigger an autoimmune response in NMOSD. (Received February 13, 2018; Accepted July 12, 2018; Published October 1, 2018).

Role of Integrin Signaling Activation on the Development of Human T Cell Leukemia Virus-1 (HTLV-1)-Associated Myelopathy/Tropical Spastic Paraparesis: Its Relationship to HTLV-1-Infected CD4+ T Cell Transmigrating Activity into the Tissues.

The main clinical feature of human T cell leukemia virus-1 (HTLV-1)-associated myelopathy/tropical spastic paraparesis (HAM/TSP) is slowly progressive spastic paraparesis with bladder dysfunction. HAM/TSP is induced by chronic inflammation in the spinal cord, mainly the lower thoracic cord. A long-standing bystander mechanism, such as the destruction of surrounding tissues by the interaction between infiltrated Th1-like, HTLV-1-infected CD4+ T cells and HTLV-1-specific CD8+ cytotoxic T cells (CTL), is probably critical for the induction of chronic inflammation. Although the HTLV-1-infected CD4+ T cells in HAM/TSP appear to play a crucial role in the initial pathogenesis of HAM/TSP, the exact mechanisms of how these cells acquire their function as the first responders in the pathogenesis of HAM/TSP still remain unresolved. Herein, we propose the importance of the activation of both outside-in signals from integrin signaling and inside-out signals for integrin signaling in the HTLV-1-infected CD4+ T cells of HAM/TSP patients.

[Case of herpes simplex encephalitis with hypersomnia and low orexin level in the cerebrospinal fluid].

A 60-year-old woman suffered from high fever (38°C) and abnormal behavior, was admitted to our hospital on the seventh day of the fever. At admission, she was stuporous, and a cerebrospinal fluid (CSF) analysis revealed pleocytosis (55/µl, monocytes). Fluid-attenuated inversion recovery (FLAIR) magnetic resonance (MR) images showed high-intensity signals in the medial temporal lobe, inferior surface of the frontal cortex, right cerebellar vermis, and left thalamus. We diagnosed herpes simplex encephalitis, based on the finding of an elevated titer of herpes simplex virus antibody in the CSF (2.90). She was started on treatment with acyclovir and steroid pulse therapy, which was followed by rapid clinical improvement. After recovering from the stupor, the patient exhibited the symptoms of hypersomnia with low orexin level in the CSF. Thus, we should bear in mind that other than consciousness disturbance, patients with herpes simplex encephalitis can also present with rare complications due to the extent of the lesions.

Animal protein intake is associated with higher-level functional capacity in elderly adults: the Ohasama study.

OBJECTIVES: To determine the association between protein intake and risk of higher-level functional decline in older community-dwelling adults. DESIGN: Prospective. SETTING: Ohasama Town, Japan. PARTICIPANTS: Residents (N = 1,007; mean age 67.4 ± 5.5) free of functional decline at baseline; follow-up was conducted for 7 years. MEASUREMENTS: Nutrient and food intakes were determined using a validated 141-item food frequency questionnaire. Participants were divided into quartiles according to intake levels of total, animal, and plant protein. Subscales of the Tokyo Metropolitan Institute of Gerontology Index of Competence subscales were used to assess higher-level functional decline. Logistic regression analysis was used to examine the future risk of higher-level functional decline in relation to protein intake, with lowest protein intake as reference. RESULTS: During the study period, 24.4% of eligible participants reported declines in higher-level functional capacity. After adjustment for putative confounding factors, men in the highest quartile of animal protein intake had significantly lower risk of higher-level functional decline than those in the lowest quartile (odds ratio (OR) = 0.41, 95% confidence interval (CI) = 0.20-0.83; P for trend .01). These associations were not seen in women (OR = 0.76, 95% CI = 0.41-1.34; P for trend .37). No consistent association was observed between plant protein intake and future higher-level functional decline in either sex. CONCLUSION: Higher protein, particularly animal protein, was associated with lower risk of decline in higher-level functional capacity in older men. Animal protein intake may be a modifiable indicator for early detection and prevention of higher-level functional decline in elderly adults.

Health behaviors as predictors for declines in higher-level functional capacity in older adults: the Ohasama study.

OBJECTIVES: To determine the characteristics of health behaviors related to higher-level functional decline in older community-dwelling adults. DESIGN: Prospective. SETTING: Ohasama Town, Japan. PARTICIPANTS: One thousand fifty residents (mean age: 67.5) free of functional decline at baseline. MEASUREMENTS: Health behaviors including smoking status, alcohol consumption, frequency of exercise, sleep duration, dietary habits (supplement use, breakfast, late-night snacking, eating regularly, and eating out), and self-rated health were obtained from a self-administered questionnaire at baseline. Higher-level functional decline was examined using the subscales of the Tokyo Metropolitan Institute of Gerontology Index of Competence. RESULTS: During the 7-year follow-up, 27.5% of eligible participants reported decline in higher-level functional capacity. After adjustment for putative confounding factors, health behaviors that were significant predictors for declines in higher-level functional capacity at the 7-year follow-up were current smoking (odds ratio (OR) = 1.58, 95% confidence interval (CI) = 1.06-2.36), sleep duration of 9 hours or longer (OR = 2.15, 95% CI = 1.49-3.11), and poor self-rated health (OR = 1.93, 95% CI = 1.40-2.67). CONCLUSION: Several modifiable health behaviors contribute to higher-level functional decline.

Disulfide-mediated apoptosis of human T-lymphotrophc virus type-I (HTLV-I)-infected cells in patients with HTLV-I-associated myelopathy/tropical spastic paraparesis.

BACKGROUND: This study was conducted to construct a basis for a therapeutic strategy against human T-lymphotropic virus type-I (HTLV-I)-associated myelopathy/tropical spastic paraparesis (HAM/TSP) using a compound that contained a disulfide moiety, prosultiamine, which is a homologue of allithiamine originally synthesized by allicin and thiamine-thiol, for the targeting of HTLV-I-infected cells. METHODS: First, we analysed the apoptotic pathway in allicin or prosultiamine treatment against an HTLV-I-infected T-cell line (HCT-1), derived from an HAM/TSP patient, by flow cytometry and western blot. Second, we evaluated the effect of targeting HTLV-I-infected cells in a prosultiamine in vitro treatment and in a clinical trial in HAM/TSP patients by quantitative PCR analysis of HTLV-I proviral load. RESULTS: Prosultiamine, like allicin, induced caspase-dependent apoptosis against HCT-1 cells. The fact that the loss of mitochondrial membrane potential was recovered in z-VAD-fmk-pretreated HCT-1 cells with prosultiamine treatment suggested that prosultiamine can induce caspase-dependent apoptosis through the mitochondrial pathway. On the basis of data showing that prosultiamine in vitro treatment against peripheral blood CD4(+) T-cells of HAM/TSP patients induced a significant decrease of HTLV-I proviral copy numbers by apoptosis of HTLV-I-infected cells, we treated six HAM/TSP patients with intravenous administration of prosultiamine for 14 days. As a result of this treatment, the copy numbers of HTLV-I provirus in peripheral blood decreased to approximately 30-50% of their pretreatment levels with some clinical benefits in all patients. CONCLUSIONS: Our results suggest that prosultiamine has the potential to be a new therapeutic tool that targets HTLV-I-infected cells in HAM/TSP.

HTLV-I production based on activation of integrin/ligand signaling in HTLV-I-infected T cell lines derived from HAM/TSP patients.

OBJECTIVE: Activity of integrin/ligand signaling leading to activation of small GTPases might regulate the efficiency of cell-to-cell spread of human T lymphotropic virus type I (HTLV-I) through the virological synapse. We compared both activity of small GTPases and involvement of integrin/ligand signaling in extracellular release of HTLV-I virions between each three HTLV-I-infected T cell lines derived from HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP) patients and from other origins as a control. METHODS: Activity of small GTPases (Rho, Rac and Cdc42) was analyzed by pull-down assay with suppressive effect of both HTLV-I production and HTLV-I tax mRNA expression by anti-integrin-blocking antibodies. RESULTS: All small GTPases were strongly activated in all cell lines derived from HAM/TSP patients, but not in control cell lines except one cell line. Treatment of all cell lines derived from HAM/TSP patients, but not all control cell lines, with anti-integrin-blocking antibodies significantly suppressed the level of HTLV-I p19 antigen in culture supernatants without downregulation of HTLV-I tax mRNA expression. CONCLUSION: Significant involvement with integrin/ligand signaling in extracellular release of HTLV-I virions in cell lines derived from HAM/TSP patients suggests that HTLV-I-infected cells in HAM/TSP patients have the potential for the efficient spread of HTLV-I to uninfected cells.

Blockade of IL-2 receptor suppresses HTLV-I and IFN-gamma expression in patients with HTLV-I-associated myelopathy/tropical spastic paraparesis.

OBJECTIVE: Th1 activation based on a high HTLV-I proviral load is one of the characteristic immunological abnormalities in the peripheral blood lymphocytes of patients with HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP). To clarify the cause of this abnormality with the potential to be one of the therapeutic targets, we analyzed the involvement of interleukin-2 (IL-2)/IL-2 receptor (IL-2R) signaling in HTLV-I and interferon-gamma (IFN-gamma), which is a representative Th1 cytokine, expression in peripheral blood CD4(+) T cells from HAM/TSP patients. PATIENTS AND METHODS: Twelve patients with HAM/TSP were included in the study. After the peripheral blood CD4(+) T cells were treated in cultures under the presence of each anti-IL-2Ralpha, beta,and gamma blocking antiboby for 48 hours, both HTLV-I p19 antigen and IFN-gamma levels in the culture supernatants were measured using ELISA methods. To check the influence on cell proliferation under these culture conditions, the numbers of viable cells were simultaneously determined by MTS assay. RESULTS: Treatment with anti-IL-2Ralpha blocking antibody, but not anti-IL-2Rbeta or anti-IL-2Rgamma blocking antibody, suppressed HTLV-I p19 antigen expression levels. In addition, treatment with all types of anti-IL-2R blocking antibodies also suppressed IFN-gamma expression levels. All of the types of anti-IL-2R blocking antibodies did not inhibit the proliferation. CONCLUSION: These results indicate that IL-2/IL-2R signaling is involved in HTLV-I and IFN-gamma expression on peripheral blood CD4(+) T cells from HAM/TSP patients, suggesting that the interruption of this signaling has therapeutic potential against HAM/TSP in patients with the focus on the down-regulation of Th1 activation based on a high HTLV-I proviral load in the peripheral blood.

Cochlear changes in patients with type 1 diabetes mellitus.

OBJECTIVE: To evaluate the effects of diabetes on cochlear elements in human beings. STUDY DESIGN AND SETTING: Twenty-six temporal bones (mean age, 37.5 years) with type 1 diabetes and 30 age-matched controls were examined by light microscopy. We compared the findings of cochlear vessels, hair cells, spiral ganglion cells, and cochlear lateral walls. RESULTS: In diabetics, the walls of vessels of the basilar membrane (P < 0.001) and vessels of the stria vascularis were (P < 0.01) significantly thicker in all turns and loss of outer hair cells (OHCs) was significantly greater in the lower basal turn (P < 0.01). Atrophy of the stria vascularis in all turns (P < 0.0001) and loss of spiral ligament cells in upper turns (P < 0.01) were significantly higher than controls. No significant difference was obtained in the number of spiral ganglion cells between groups. CONCLUSION: This study suggests that type 1 diabetes mellitus can cause cochlear microangiopathy and subsequently degeneration of cochlear lateral walls and OHCs.

Large deletion involving the 5'-UTR in the spastin gene caused mild phenotype of autosomal dominant hereditary spastic paraplegia.

Hereditary spastic paraplegia (HSP) due to mutations in the spastin gene (SPG4) located to 2p22-p21 is the most common form of autosomal dominant (AD) HSP. We performed PCR-based direct sequencing of SPG4, followed by a linkage analysis and subsequent Southern blot analysis in large Japanese kindred where 20 of 33 members were evaluated neurologically, and consequently 6 were affected with HSP. Clinical evaluation showed that the mean age at disease onset of the patients was older and the disability was less severe than those of previously reported typical patients with SPG4 mutations. Direct sequencing of genomic DNA and RT-PCR product did not show a SPG4 mutation despite of a strong linkage to the SPG4 locus at 2p. Southern blot analysis suggested a deletion involving the 5'-UTR of SPG4. Further sequence analysis confirmed a heterozygous 2307-bp deletion spanning from the 5'-UTR to intron 1 of SPG4. The results suggested that transcription of the mutated allele starts from an authentic initiation site, but lacks an authentic translational start site of exon 1 because of a deficient splice donor site and coding region. The abnormal transcripts may result in rapid RNA decay. The novel refractory mutation we identified widens the spectrum of SPG4 mutations. These findings suggest that structural genomic abnormalities of SPG4 are more frequent than expected, and this explains previously reported cases more feasibly in which SPG4 mutations were failed to be identified but the disease was linked to 2p.

Involvement of p38 MAPK signaling pathway in IFN-gamma and HTLV-I expression in patients with HTLV-I-associated myelopathy/tropical spastic paraparesis.

We analyzed the relationship between the expression of interferon (IFN)-gamma and HTLV-I p19 antigen and activation of p38 mitogen-activated protein kinase (p38 MAPK) in two HTLV-I-infected T cell lines derived from two patients (HCT-1 and HCT-4) with HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP), and three HTLV-I-infected T cell lines derived from three patients with adult T cell leukemia (ATL). Expression of phosphorylated (activated)-p38 MAPK was markedly increased concomitant with high levels of both IFN-gamma and HTLV-I p19 antigen expression in both HCT-1 and HCT-4 compared with cell lines derived from ATL patients. Treatment with SB203580, a specific inhibitor of p38 MAPK, suppressed IFN-gamma and HTLV-I p19 antigen expression levels in HCT-1, HCT-4 and peripheral blood CD4(+) T cells of HAM/TSP patients. These findings strongly suggest that activation of p38 MAPK signaling pathway is involved in the up-regulation of IFN-gamma expression with high HTLV-I proviral load in HAM/TSP patients.

Increased mRNA expression of Th1-cytokine signaling molecules in patients with HTLV-I-associated myelopathy/tropical spastic paraparesis.

Expression of inflammatory cytokines derived from Th1 cell population is increased in patients with human T-lymphotropic virus type I (HTLV-I)-associated myelopathy/tropical spastic paraparesis (HAM/TSP). It has been shown that cytokine signaling molecules, including transcription factors T-bet and GATA-3, interleukin-12 receptor beta 2 (IL-12R beta 2) and suppressors of cytokine signaling (SOCS), such as SOCS1, are important in differentiation of naive T cells into Th1 helper T cells. To assess the immunological status from the stand-point of cytokine signaling in patients with HAM/TSP, we analyzed mRNA expression of these cytokine signaling molecules in peripheral blood mononuclear cells using quantitative RT-PCR. Twenty-eight HAM/TSP patients, nine HTLV-I-infected individuals without HAM/TSP and twenty-two HTLV-I-uninfected individuals were included in this study. Expression of T-bet, GATA-3, IL-12R beta 2 and SOCS1 was significantly increased in HAM/TSP patients in comparison with HTLV-I-uninfected individuals. In contrast, expression of SOCS3, a marker for Th2 cells, was significantly decreased in HTLV-I-infected individuals. These results indicate that HAM/TSP patients are associated with increased Th1 and decreased Th2 cytokine signaling activities.