Impact of Energy Malnutrition on Exacerbation Hospitalization in Patients with Chronic Obstructive Pulmonary Disease: Retrospective Observational Study.

Nutritional disorders in patients with chronic obstructive pulmonary disease (COPD) are associated with cachexia, sarcopenia, and weight loss. In particular, weight loss is a prognostic factor in COPD independent of pulmonary function, and energy malnutrition is a contributing factor. Frequent exacerbation hospitalization is also a prognostic factor for COPD patients. The impact of energy malnutrition on adverse events such as exacerbation hospitalization is unknown, and this study aimed to investigate that. We included 163 male subjects with COPD. Respiratory quotient (RQ), an index of energy malnutrition, was calculated by expiratory gas analysis using an indirect calorimeter. RQ <0.85 was categorized as the energy malnutrition group. Kaplan-Meier analysis was used to compare the hospitalization avoidance rate between the with and without energy malnutrition groups. Independent factors associated with exacerbation hospitalization were evaluated by Cox regression analysis. We finally analyzed data from 56 selected subjects (median age: 74 y). The exacerbation hospitalization rate was significantly higher in the energy malnutrition group. Fifty percent of the energy malnutrition group was hospitalized for an exacerbation, and the median hospitalization avoidance time was 701 d. In Cox regression analysis (adjusted for age, BMI, mMRC dyspnea scale score, %FEV1, and 6-min walk test), energy malnutrition was an independent factor associated with exacerbation hospitalization (HR 4.14, 95% CI 1.13-15.1, p=0.03). Energy malnutrition may be the risk factor for exacerbation hospitalization. Energy malnutrition may be an early nutritional disorder and early detection and intervention may reduce exacerbation hospitalizations.

CHARACTERISATION OF RADIOACTIVE CAESIUM IN SEDIMENTS AT THE NOGAWA RIVER, JAPAN.

The TEPCO Fukushima Daiichi Nuclear Power Plant accident that occurred in March 2011 resulted in the release of radioactive caesium into the environment. The radioactive caesium has been detected in the Tama River watershed. Previous investigations have shown that the concentration of radioactive caesium in sediment was relatively high in the Nogawa River. In this study, the relationship between the concentration of radioactive caesium in the sediment and the sediment characteristics was investigated. We found that 137Cs concentration in the tributary sediment has difficulty migrating downstream, while exhibiting a strong correlation with the amount of organic matter and a correlation with the clay, silt layer. Based on the results, we inferred that 137Cs is deposited together with the organic matter and clay, silt layer in the sediment and migrates at a slower pace than that in the mainstem.

Synergistic effect of hypertension and smoking on the total small vessel disease score in healthy individuals: the Kashima scan study.

The total cerebral small vessel disease (SVD) score is a proposed comprehensive index of SVD severity in the brain. However, data on lifestyle-related risk factors affecting SVD scores are limited. We conducted a cross-sectional study with 858 neurologically healthy adults who underwent brain magnetic resonance imaging (MRI). Information on clinical and lifestyle-related risk factors was obtained from health screenings. The SVD score (0-4) was calculated from the presence of lacunes, cerebral microbleeds, moderate to severe white matter lesions, and basal ganglia perivascular spaces on MRI. Subjects were divided into two groups by SVD score; potential risk factors and their joint effects in the two groups were assessed by logistic regression. Biologic interactions were estimated using the synergy index. After adjustment for possible confounders, the adjusted odds ratio for moderate to severe SVD scores (SVD score ≥ 2) was 1.12 (95% confidence interval (CI) 1.08-1.16) for age per year, 1.33 (95% CI 1.02-1.74) for body mass index per standard deviation, 3.39 (95% CI 1.90-6.03) for hypertension, 2.31 (95% CI 1.14-4.69) for diabetes, and 2.35 (95% CI 1.10-5.02) for smoking. Hypertension and current smoking had a synergistic effect on the risk of moderate to severe SVD (OR 10.59, 95% CI 3.97-28.3; synergy index 4.03, 95% CI 1.17-28.30), and the combination of hypertension and diabetes had an additive effect on the risk of moderate to severe SVD (OR 9.48, 95% CI 3.80-23.66; synergy index 2.12, 95% CI 0.68-6.67). Therefore, combined strategies for managing hypertension, smoking, and diabetes may be effective for preventing SVD.

Total Small Vessel Disease Score in Neurologically Healthy Japanese Adults in the Kashima Scan Study.

Objective We explored the association between the total small vessel disease (SVD) score obtained with magnetic resonance imaging and risk factors and outcomes in the Japanese population. Methods The presence of SVD features, including lacunes, cerebral microbleeds, white matter changes, and basal ganglia perivascular spaces on MRI, was summed to obtain a "total SVD score" (range 0-4). Ordinal and multinomial logistic regression analyses were performed to investigate the association of higher total SVD scores with vascular risk factors, the Mini-Mental State Examination (MMSE) score, and cerebral atrophy. Results We included 1,451 neurologically healthy adults (mean age, 57.1 years; 47% male). A multivariate ordinal logistic regression analysis showed that the total SVD score was associated with aging, hypertension, blood pressure (BP), diabetes mellitus, MMSE score, and deep cerebral atrophy, but the equal slopes assumption between scores did not hold. A multivariate multinomial logistic regression analysis (total SVD score 0=reference) showed that aging, hypertension, and BP were positively associated with scores of 1, 2, or ≥3. These effects, presented as odds ratios (ORs), increased as the score increased and were strongest with a score of ≥3 [aging (per 10-year increment), OR 4.00, 95% confidence interval (CI) 2.47-6.46; hypertension, OR 5.68, 95% CI 2.52-12.80; systolic BP (per standard deviation increase), OR 1.96, 95% CI 1.41-2.74, respectively]. Diabetes mellitus and deep cerebral atrophy tended to be associated with the SVD scores. The MMSE score showed no consistent associations. Conclusion The total SVD score may be a promising tool for indexing SVD, even in the Japanese population.

Basal ganglia cerebral microbleeds and global cognitive function: the Kashima Scan Study.

BACKGROUND: We previously showed that global cognitive function was associated with deep or infratentorial (D/I) cerebral microbleeds (CMBs) in a Japanese healthy cohort. We continually recruited participates and performed further investigation to focus on the impact of different distributions of D/I CMBs on gradient-echo magnetic resonance imaging on global cognitive function. METHODS: A total of 1392 subjects including subjects without CMBs (n = 1335), with D/I CMBs limited to the basal ganglia (BG; BG group, n = 33), thalamus (thalamus group, n = 14), and infratentorial area (infratentorial group, n = 10) were included in analyses. Subjects with strictly lobar CMBs (n = 43) were excluded, but subjects in the BG, thalamus, and infratentorial groups could also have lobar CMBs. The mini-mental state examination (MMSE) was administered to determine global cognitive function; scores less than 27 or more than 1.5 standard deviations (SDs) below the age-education-related mean were regarded as impaired. RESULTS: In the multivariable logistic regression analyses, hypertension and severe white matter hyperintensities were associated with the BG group and the thalamus group. In multivariable logistic regression analysis of the association between D/I CMBs classification and impaired MMSE score, only the BG group consistently displayed associations with both MMSE score less than 27 (odds ratio [OR], 5.96; 95% confidence interval [CI], 2.08-17.09) and MMSE score more than 1.5 SDs below the age-education-related mean (OR, 3.34; 95% CI, 1.24-8.99). In the BG group, adjusted mean scores of total MMSE and "attention and calculation" were lower compared with subjects without CMBs. CONCLUSIONS: In our study of D/I CMBs, only BG CMBs have strong association with global cognitive function. This association was independent of CMBs in other location.

Norms of the Mini-Mental state Examination for Japanese subjects that underwent comprehensive brain examinations: the Kashima Scan Study.

OBJECTIVE: The distribution of the Mini-Mental State Examination (MMSE) scores by age and educational level was investigated in subjects that underwent comprehensive brain examinations. METHODS: This cross-sectional study included 1,414 adults without neurological disorders who underwent health-screening tests of the brain, referred to as the "Brain Dock," in our center. The MMSE scores were compared between age groups (40-44, 45-49, 50-54, 55-59, 60-64, 65-69, or ≥70 years) and educational levels [the low education level group (6-12 years) and the high education level group (≥13 years)]. RESULTS: The median age was 59 years, and 763 (54%) were women. There was no significant difference in the MMSE total score between women and men. The stepwise method of the multiple linear regression analysis confirmed that a higher age [ß value, -0.129; standard error (S.E.), 0.020; p<0.001], low education level (6-12 years) (ß value, -0.226; S.E., 0.075; p=0.003), and women (ß values, 0.148; S.E., 0.066; p=0.024) was significantly associated with decreased MMSE score. In general, both the percentile scores and mean scores decreased with aging and were lower in the low education level group than in the high education level group. The degree of decrement in scores with age was stronger in the low education level group than in the high education level group. CONCLUSION: The provided data for age- and education-specific reference norms will be useful for both clinicians and investigators who perform comprehensive brain examinations to assess the cognitive function of subjects.

Topography and associations of perivascular spaces in healthy adults: the Kashima scan study.

OBJECTIVE: We investigated whether the topography of MRI-visible perivascular spaces (PVS) is associated with markers of specific underlying small vessel disease, including cerebral microbleed (CMB) distribution, in neurologically healthy adults. METHODS: We analyzed baseline data of an ongoing Japanese population-based cohort study. PVS were rated in the basal ganglia (BG-PVS) and centrum semiovale (CSO-PVS) on axial T2-weighted MRI using a validated rating scale (score 0-4). BG-PVS degree was classified as low (score <2) or high (score ≥2). CSO-PVS degree was classified as low (score <3) or high (score ≥3). Independent demographic, clinical, and imaging factors for high degree of BG-PVS and CSO-PVS were investigated. RESULTS: A total of 1,575 neurologically healthy adults were included (mean age 57.1 years, SD 9.7; 47% male). In multivariable analyses, high degree of BG-PVS (n = 212, 14%) was associated with deep or infratentorial CMBs (odds ratio [OR] 2.77, 95% confidence interval [CI] 1.62-4.74), a marker of hypertensive arteriopathy; by contrast, high degree of CSO-PVS (n = 357, 23%) was associated with strictly lobar CMBs (OR 2.49, 95% CI 1.35-4.61), which share risk factors with cerebral amyloid angiopathy. Both high degree of BG-PVS and CSO-PVS were associated with hypertension (OR 2.03, 95% CI 1.46-2.82 and OR 1.39, 95% CI 1.07-1.81, respectively), lacunes (OR 3.35, 95% CI 1.92-5.86; OR 1.83 95% CI 1.08-3.08), and severe white matter hyperintensities (OR 2.17, 95% CI 1.42-3.31; OR 1.35, 95% CI 0.93-1.96), but these associations were stronger for high degree of BG-PVS. CONCLUSIONS: In a neurologically healthy cohort, the associations of PVS differ according to their topography. PVS distribution may be useful for the early detection and classification of small vessel disease.

Microvascular decompression surgery for vertebral artery compression of the medulla oblongata: 3 cases with respiratory failure and/or dysphagia.

BACKGROUND: It is well known that brainstem dysfunction may be caused by vascular compression of the medulla oblongata (MO). However, only a limited number of reports have found microvascular decompression (MVD) surgery to be an effective treatment for symptomatic patients with MO dysfunction, such as essential hypertension, pyramidal tract signs, dysphagia, and respiratory failure. CASE DESCRIPTION: This report describes 3 patients with vertebral artery compression of MO who presented with respiratory failure and/or dysphagia. MVD surgery using the transcondylar fossa approach was effective in relieving patient symptoms. CONCLUSIONS: Although the pathogenic mechanisms of symptomatic vertebral artery compression of MO remain unclear, we should recognize that MVD surgery is effective for selected patients with brainstem dysfunction. The transcondylar fossa approach and the stitched sling retraction technique are appropriate in MVD surgery to relieve vertebral artery compression of MO.

[A case of migraine with aura attenuated by transcatheter closure of atrial septal defect].

A 38-year old man with an 8-year history of migraine with subclinical abnormal brain lesions on MRI was admitted to our hospital. His migraine attacks followed visual disturbance or dysarthria. Brain MRI revealed old infarctions restricted to the posterior circular region. On transesophageal cardiography, an atrial septal defect (ASD) was detected, and a bubble study showed an immediate appearance of many bubbles in the left atrium via ASD without Valsalva maneuver. The bubble study on transcranial-color-flow imaging also detected micro-embolic signals at the left vertebral artery and the left middle cerebral artery without Valsalva maneuver. Since paradoxical embolism via ASD was highly suspected and Qp/Qs was more than 1.5, transcatheter closure of ASD using AMPLATZER(®) Septal Occluder was performed. At a 2-year follow up, no recurrence of either migraine or infarction was found. This case indicates the relevance of right-to-left shunt to migraine with aura, as well as the usefulness of transcatheter closure of ASD using AMPLATZER(®) Septal Occluder for treatment of migraine with aura.

[A case of superficial siderosis treated with intravenous and oral hemostatic drugs].

A 39-year-old man suffering from progressive dysarthria, gait disturbance, and sensorineural deafness for 2 years was admitted to our hospital. He scored 28 points on the mini-mental state examination. He had previously undergone surgery at 24 years and 39 years of age for a cerebellar tumor (pilocytic astrocytoma). Superficial siderosis (SS) was diagnosed based on bloody cerebrospinal fluid (CSF) and the findings of T2-weighted head MRI that revealed marginal hypointensity of the surface of the cerebellum, brainstem, and cerebral cortex. After intravenous infusion and the oral use of hemostatic drugs (carbazochrome, tranexamic acid), the CSF became watery clear and his condition improved. Hemostatic drug therapy should be considered for SS.

[Case of cheiro-oral syndrome with a bilateral perioral sensory disturbance caused by unilateral pontine tegmental hemorrhage].

A case of a small left pontine tegmental hemorrhage that presented as cheiro-oral syndrome with a bilateral perioral sensory disturbance is described. An 83-year-old man suddenly developed numbness in his bilateral perioral area and right hand. Head CT and MRI (T(2)*-weighted image) revealed a small left pontine tegmental hemorrhage. The patient was diagnosed as having cheiro-oral syndrome with bilateral perioral sensory disturbance, probably due to unilateral pontine tegmental hemorrhage. All residual symptoms disappeared within a month.In the present case, the following clinicopathological hypothesis was considered. The hematoma located in the left pontine tegmentum impaired the sensory fibers from the contralateral medial lemniscus (from the right hand) and the ventral trigeminothalamic tract (from the right perioral region). In addition, the ipsilateral trigeminothalamic tract (from the left perioral region) was also impaired. It is important to know that a small unilateral lesion in the brainstem (especially the pons) can cause cheiro-oral syndrome with a bilateral perioral sensory disturbance, and a small brainstem hematoma is the most frequent etiology of this disease.

Stroke scale items associated with neurologic deterioration within 24 hours after recombinant tissue plasminogen activator therapy.

It is unclear when and which neurologic deficits should be examined within 24 hours after intravenous recombinant tissue plasminogen activator (rt-PA) therapy for acute ischemic stroke. Relationships between serial changes in National Institutes of Health Stroke Scale (NIHSS) subscores and neurologic deterioration (ND) within the first 24 hours after therapy were investigated in 43 consecutive patients. The NIHSS score was measured by neurologists 28 times within 24 hours after therapy. Assessments of subscores associated with ND, defined as the first change 4 or more points from baseline, were performed at 15 minutes (most frequent time of the first ND), 120 minutes (median time of the first ND), and 24 hours after therapy. Seventeen of 43 patients (age range, 55-94 years) showed ND. Of the NIHSS subscores, increases in scores for loss of consciousness (15 minutes, P = .001; 120 minutes, P = .026; 24 hours, P = .018) and motor limbs total (15 minutes, P = .014; 120 minutes, P = .031) were related to deterioration. Items such as questions, gaze, visual fields, ataxia, language, dysarthria, and extinction/inattention were not related to deterioration at any time. In conclusion, ND of ischemic stroke patients treated with intravenous rt-PA therapy was frequently seen within 120 minutes after therapy. Items such as loss of consciousness and motor limbs total may be considered indices for monitoring neurologic deficits after therapy.

Distributional impact of brain microbleeds on global cognitive function in adults without neurological disorder.

BACKGROUND AND PURPOSE: Brain microbleeds (MBs) are considered to be associated with cognitive decline and can be pathologically and topographically classified as cerebral amyloid angiopathy-related (located in lobar regions) and hypertensive microangiopathy-related (located in deep regions). We examined whether different effects on global cognitive function might be seen with different distributions of MBs. METHODS: A total of 1279 adults without neurological disorders were studied prospectively. Subjects were divided into 4 groups: without-MBs group; lobar group; deep group; and with in both areas (diffuse group). The Mini-Mental State Examination was administered to determine global cognitive functions, with scores<27 regarded as subnormal. RESULTS: MBs were detected in 98 subjects (8%): 36 subjects (3%) classified as lobar group, 48 subjects (4%) as deep group, and 14 subjects (1%) as diffuse group. Subnormal scores were found in 76 subjects (5.9%), associated with age, education, hypertension, severe white matter hyperintensities, and distribution and number of MBs. In the final model of logistic regression analysis, the deep group (OR, 2.79; 95% CI, 1.14-6.79) was associated with subnormal scores, whereas the lobar group (OR, 0.77; 95% CI, 0.17-3.44) was not. Trend for the diffuse group did not reach the level of significance (OR, 5.01; 95% CI, 0.88-28.41). These trends were also seen in analysis using another cut-off point for subnormal score. Scores for total Mini-Mental State Examination and attention and calculation were significantly lower in the deep group and the diffuse groups compared with the without-MBs group. CONCLUSIONS: This Japanese cross-sectional study demonstrated that MB-related global cognitive dysfunction seems to occur based on hypertensive pathogenesis rather than on cerebral amyloid angiopathy.

Identification of astrocyte-derived immune suppressor factor that induces apoptosis of autoreactive T cells.

In experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis, apoptosis of T cells is mainly seen at inflammation sites of the central nervous system (CNS). Cumulative data suggests that astrocytes might render T cells susceptible to induction of apoptotic cell death. We observed that apoptotic cell death of proteolipid protein (PLP)-reactive T cells was induced by an interferon (IFN)-γ-treated astrocyte cell line. In this study, we have identified and cloned the genes derived from the IFN-γ-treated astrocyte cell line that induce apoptosis of autoreactive T cells. We created subtraction cDNA libraries from the IFN-γ-treated astrocyte cell line and obtained 100 positive clones. After screening of subtracted cDNAs, we found two candidate genes that induced apoptosis of the PLP-reactive T cell line. The first is a previously unknown gene of 726 base pairs that we named astrocyte-derived immune suppressor factor (AdIF). It contained an open reading frame encoding a polypeptide of 228 amino acids. The second was SPARC/osteonectin, a multifunctional glycoprotein secreted in the extracellular matrix. AdIF protein was found at the inflammatory sites of the EAE brain, and bound to the surface of CD4(+) T cells. Purified recombinant AdIF protein inhibited the proliferation of activated PLP-reactive CD4(+) T cells and induced their apoptosis in vitro. Intravenous administration of recombinant AdIF protein to mice with in which acute EAE was induced prevented the incidence of EAE and suppressed the symptoms. The newly discovered molecule AdIF may render auto-reactive T cells susceptible to the induction of apoptotic cell death and could potentially be a new therapeutic agent for multiple sclerosis.

Marked cerebral atrophy is correlated with kidney dysfunction in nondisabled adults.

The relationship between kidney dysfunction, such as chronic kidney disease (CKD), and brain morphology has attracted increasing attention, but the association between kidney dysfunction and cerebral atrophy has yet to be determined. The purpose of this study was to clarify the relationship between kidney function and a substantial degree of cerebral atrophy. A total of 610 consecutive Japanese adults without neurological disorders who had undergone health screening tests of the brain were studied prospectively. Magnetic resonance imaging was performed using a 1.5-T scanner. Using a computer-assisted processing system, the percentage of cerebrum atrophy (%Cerebrum atrophy) was calculated as an index of cerebral atrophy. Atrophy was defined as >2 s.d.s below the mean %Cerebrum atrophy. The glomerular filtration rate (GFR) was estimated using the revised equations for estimated GFR from serum creatinine in Japan. Kidney function variables included the GFR value and the prevalence of subjects with GFR <60 ml min⁻¹ per 1.73 m². Cerebral atrophy was found in 25 (4.1%) cases. Univariate analysis showed that age, male sex, hypertension, each kidney function variable, white matter hyperintensities and lacunae were associated with cerebral atrophy. On logistic regression analysis, GFR (odds ratio (OR), 0.64; 95% confidence interval (CI), 0.42-0.98) and GFR <60 ml min⁻¹ per 1.73 m² (OR, 5.93; 95% CI, 1.82-19.27) were significantly associated with cerebral atrophy. On sub-analysis, GFR <60 ml min⁻¹ per 1.73 m² was significantly associated with cortical atrophy (OR, 3.23; 95% CI, 1.15-9.11). Decreased GFR was significantly associated with cerebral atrophy, indicating that treatment of CKD may control age-related degenerative processes of the brain.

Incidence and clinical significances of human T-cell lymphotropic virus type I-associated myelopathy with T2 hyperintensity on spinal magnetic resonance images.

OBJECTIVE: To clarify the incidence and clinical significance of HTLV-I associated myelopathy/tropical spastic paraparesis (HAM/TSP) showing T2 hyperintensity in the spinal cord on magnetic resonance images (MRI). PATIENTS AND METHODS: We reviewed the spinal cord MRI of 38 HAM/TSP patients and analyzed them in relation to clinical and laboratory findings. Analyzed data were: age at onset, disease duration, disability status, responsiveness to interferon therapy, brain abnormalities on MRI, serum anti-HTLV-I titers, and cerebrospinal fluid (CSF) findings. RESULTS: MRI findings of the spinal cord were classified into 3 types, "normal" (n=22, 57.9%), "atrophy" (n=13, 34.2%) and "T2-hyperintensity" (n=3, 7.9%). Patients in the normal and atrophy types showed slowly progressive paraparesis. Significant differences were not found between the normal and atrophy types in any clinical or laboratory data, including disease duration, disability status and responsiveness to interferon-alpha therapy. Meanwhile, all patients showing T2-hyperintensity had severe paraparesis of a rapid progressive nature, with CSF IgG elevation. CONCLUSION: HAM/TSP with T2-hyperintensity on spinal MRI shows a rapid progressive clinical course with severe motor impairment. The incidence of this malignant form of HAM/TSP is estimated to be around 7.9%.

Brain microbleeds and global cognitive function in adults without neurological disorder.

BACKGROUND AND PURPOSE: Increasing attention has been paid to associations between cognitive dysfunction and brain microbleeds (MBs). Because all previous studies have investigated patients with neurological disorders, we examined subjects without neurological disorder in order to clarify pathogenic relationships. METHODS: A total of 518 consecutive adults without neurological disorder who had undergone health-screening tests of the brain were studied prospectively. Gradient-echo T2*-weighted MRI using a 1.5-T system was used to detect MBs. The Mini-Mental State Examination (MMSE) was administered to determine cognitive functions. MMSE scores <27 or >1.5 SDs below the age-related mean were regarded as subnormal. RESULTS: MBs were found in 35 subjects (6.8%). MMSE score <27 was found in 25 subjects (4.8%), with MMSE score >1.5 SDs below the age-related mean in 34 subjects (6.6%). Univariate analysis showed presence and number of MBs, short duration of education, and severe white matter hyperintensities as significantly associated with subnormal scores. In logistic regression analysis, presence of MBs (odds ratio [OR], 5.44; 95% CI, 1.83 to 16.19) and number of MBs (OR, 1.32; 95% CI, 1.04 to 1.68) still displayed significant associations with MMSE score <27. Logistic regression analysis revealed a significant relationship between presence (OR, 3.93; 95% CI, 1.44 to 10.74) and number (OR, 1.26; 95% CI, 1.01 to 1.59) of MBs and MMSE score >1.5 SDs below the age-related mean. Among MMSE subscores, "attention and calculation" was significantly lower in MB-positive subjects (P=0.017). CONCLUSIONS: MBs appear to be primarily associated with global cognitive dysfunction.

Human astrocytes express 14-3-3 sigma in response to oxidative and DNA-damaging stresses.

The 14-3-3 protein family consists of seven isoforms, most of which are expressed abundantly in neurons and glial cells, although the sigma isoform, a p53 target gene originally identified as an epithelium-specific marker, has not been identified in the human central nervous system. Here, we show that human astrocytes in culture expressed 14-3-3sigma under stress conditions. By Western blot, the expression of 14-3-3sigma, p53 and p21 was coordinately upregulated in astrocytes following exposure to hydrogen peroxide, 4-hydroxy-2-nonenal (4-HNE) or etoposide, a topoisomerase II inhibitor. 14-3-3sigma was induced by treatment with 5-aza-2'-deoxycytidine, suggesting a hypermethylated status of the gene promoter in astrocytes. In vivo, a small subset of hypertrophic reactive astrocytes, often showing a multinucleated morphology, expressed 14-3-3sigma in active demyelinating lesions of multiple sclerosis (MS) and ischemic lesions of cerebral infarction, where the expression of 4-HNE and 8-hydroxy-2'-deoxyguanosine was enhanced in reactive astrocytes. Microarray analysis of etoposide-treated astrocytes verified upregulation of p53-responsive genes and concurrent downregulation of mitotic checkpoint-regulatory genes. These observations suggest that 14-3-3sigma might serve as a marker of oxidative and DNA-damaging stresses inducing the mitotic checkpoint dysfunction in reactive astrocytes under pathological conditions.

Microarray analysis identifies a set of CXCR3 and CCR2 ligand chemokines as early IFNbeta-responsive genes in peripheral blood lymphocytes in vitro: an implication for IFNbeta-related adverse effects in multiple sclerosis.

BACKGROUND: A substantial proportion of multiple sclerosis (MS) patients discontinue interferon-beta (IFNbeta) treatment due to various adverse effects, most of which emerge at the early phase after initiation of the treatment and then diminish with time. At present, the molecular mechanism underlying IFNbeta-related adverse effects remains largely unknown. The aim of this study is to identify a comprehensive list of early IFNbeta-responsive genes (IRGs) in peripheral blood mononuclear cells (PBMC) that may play a key role in induction of adverse effects. METHODS: Total RNA of PBMC exposed to 50 ng/ml recombinant human IFNbeta for 3 to 24 hours in vitro was processed for cDNA microarray analysis, followed by quantitative real-time RT-PCR analysis. RESULTS: Among 1,258 genes on the array, IFNbeta elevated the expression of 107 and 87 genes, while it reduced the expression of 22 and 23 genes at 3 and 24 hours, respectively. Upregulated IRGs were categorized into conventional IFN-response markers, components of IFN-signaling pathways, chemokines, cytokines, growth factors, and their receptors, regulators of apoptosis, DNA damage, and cell cycle, heat shock proteins, and costimulatory and adhesion molecules. IFNbeta markedly upregulated CXCR3 ligand chemokines (SCYB11, SCYB10 and SCYB9) chiefly active on effector T helper type 1 (Th1) T cells, and CCR2 ligand chemokines (SCYA8 and SCYA2) effective on monocytes, whereas it downregulated CXCR2 ligand chemokines (SCYB2, SCYB1 and IL8) primarily active on neutrophils. CONCLUSION: IFNbeta immediately induces a burst of gene expression of proinflammatory chemokines in vitro that have potential relevance to IFNbeta-related early adverse effects in MS patients in vivo.

Rapid identification of 14-3-3-binding proteins by protein microarray analysis.

The 14-3-3 protein family consists of acidic 30-kDa proteins composed of seven isoforms in mammalian cells, expressed abundantly in neurons and glial cells of the central nervous system (CNS). The 14-3-3 isoforms form a dimer that acts as a molecular adaptor interacting with key signaling components involved in cell proliferation, transformation, and apoptosis. Until present, more than 300 proteins have been identified as 14-3-3-binding partners, although most of previous studies focused on a limited range of 14-3-3-interacting proteins. Here, we studied a comprehensive profile of 14-3-3-binding proteins by analyzing a high-density protein microarray using recombinant human 14-3-3 epsilon protein as a probe. Among 1752 proteins immobilized on the microarray, 20 were identified as 14-3-3 interactors, most of which were previously unreported 14-3-3-binding partners. However, 11 known 14-3-3-binding proteins, including keratin 18 (KRT18) and mitogen-activated protein kinase-activated protein kinase 2 (MAPKAPK2), were not identified as a 14-3-3-binding protein. The specific binding to 14-3-3 of EAP30 subunit of ELL complex (EAP30), dead box polypeptide 54 (DDX54), and src homology three (SH3) and cysteine rich domain (STAC) was verified by immunoprecipitation analysis of the recombinant proteins expressed in HEK293 cells. These results suggest that protein microarray is a powerful tool for rapid and comprehensive profiling of 14-3-3-binding proteins.