The pentameric hydrocoel lobes organize adult pentameral structures in a sea cucumber, Apostichopus japonicus.

Despite being one of the bilaterians, the body plan of echinoderms shifts from bilateral symmetry to five-fold radial, or pentaradial symmetry during embryogenesis or their metamorphosis. While the clarification of the developmental mechanism behind this transition will be a basis for understanding their unique body plan evolution, it is still poorly understood. With this regard, the hydrocoel, a mesodermal coelom formed on the left side of bilateral larva, would be a clue for understanding the mechanism as it is the first pentaradial structure that appears before metamorphosis and develops into the water vascular system of adults. By analyzing the development of a sea cucumber, Apostichopus japonicus, we found that the hydrocoel expresses genes related in muscle and neural formation such as myosin heavy chain, tropomyosin, soxC, and elav, implying that cells of the hydrocoel contributes to muscle and neural structures in the adult. Furthermore, ablation of one of the hydrocoel lobes led to incomplete development of adult pentameral structures. The ablation of primary hydrocoel lobes resulted in the reduction in tentacles and the ablation of secondary hydrocoel lobes resulted in the reduction in water vascular canals and nerve cords. Our findings suggest that the hydrocoel lobes may serve as a potential organizing center for establishing the pentaradial body plan in echinoderms.

[Transient increase of intraocular pressure with primary open angle glaucoma patients associated with warm and cold seasons after long waiting time in an ophthalmology clinic].

PURPOSE: To retrospectively evaluate the influence of long waiting time on intraocular pressure (IOP) with primary open angle glaucoma (POAG) patients in the waiting room of an ophthalmology clinic. PATIENTS AND METHOD: We reviewed 74 consecutive patients with POAG who visited at Kaimeido Ophthalmic and Dental Clinic within the past 8 years without change of glaucoma medications, 34 men and 40 woman, average age +/- standard deviation) 67.9 +/- 13.4 years. Right eye data of all patients were enrolled and IOP was split into 4 groups, short waiting time (< 30 min ; SH30) and long waiting time (> or = 60 min; LH60) in the hot (> or = 15 degrees C) season. Short waiting time (SC30) and long waiting time (LC60) in the cold (< or = 5 degrees C) season; according to the average outdoor air temperature in Sapporo Japan. All groups were compared each other (paired t test). RESULTS: There was a significant difference between SH30 (14.6 2.4 mmHg) and LH60 (14.9 +/- 2.4 mmHg; p = 0.038), between SC30 (14.6 +/- 2.4 mmHg) and LC60 (15.5 +/- 2.6 mmHg; p = 3.45 x 10(-7)) and between LH60 and LC60 (p = 0.00079). On the other hand, there was no significant difference between SH30 and SC30 (p = 0.89). CONCLUSION: This retrospective study suggests that patients with POAG show a transient IOP elevation after longer waiting time, especially during the cold season.

Exendin-4 promotes the membrane trafficking of the AMPA receptor GluR1 subunit and ADAM10 in the mouse neocortex.

Glucagon-like peptide-1 (GLP-1) is a novel treatment modality for type 2 diabetes mellitus. However, GLP-1 has been suggested as a therapeutic target for Alzheimer's disease (AD). In rodent studies, GLP-1 reduces amyloid beta (Aß) and facilitates synaptic plasticity. Therefore, in the present study, we investigated how GLP-1 facilitates synaptic plasticity and reduces the Aß in vivo. Exendin-4, a GLP-1 receptor agonist that can cross the blood brain barrier, was subcutaneously administered to adult mice. We then extracted the total and the plasma membrane proteins from the mouse neocortex. Exendin-4 significantly increased the phosphorylation level of cAMP response element-binding protein (CREB). Consistently, the expression level of brain-derived neurotrophic factor (BDNF), a transcriptional target of CREB, was increased. Furthermore, exendin-4 increased the membrane protein level of the AMPA receptor GluR1 subunit and postsynaptic density protein-95 (PSD-95), whereas GluR2 was unaffected. These exendin-4-dependent increases in membrane GluR1, total PSD-95 and BDNF were abrogated by pretreatment with temozolomide (TMZ), a DNA-alkylating agent, indicating that these alterations were dependent on exendin-4-induced transcriptional activity. In addition, we found that exendin-4 increased the level of the α-C terminal fragment (α-CTF) of amyloid precursor protein (APP). Furthermore, protein levels of both mature and immature ADAM10, the α-secretase of APP in the plasma membrane, were increased, whereas the total mature and immature ADAM10 levels were unchanged. These exendin-4-dependent increases in α-CTF and ADAM10 were not affected by TMZ. These findings suggested that GLP-1 facilitates the GluR1 membrane insertion through CREB activation and increases α-secretase activity through ADAM10 membrane trafficking. Upregulation of GluR1 and ADAM10 at the plasma membrane were also observed in mice with intracerebroventricular administration of Aß oligomer, indicating that a part of benefit of exendin-4 against AD may depend on the GluR1 and ADAM10 membrane trafficking.

Methicillin-resistant Staphylococcus aureus carriage among veterinary staff and dogs in private veterinary clinics in Hokkaido, Japan.

To explore the prevalence and molecular characteristics of methicillin-resistant Staphylococcus aureus (MRSA) in veterinary medical practices, MRSA carriage was tested among 96 veterinarians (Vets), 70 veterinary technicians (VTs) and 292 dogs with which they had contact at 71 private veterinary clinics (VCs) in Hokkaido, Japan. MRSA isolates were obtained from 22 Vets [22.9%] and 7 VTs [10%]. The prevalence of MRSA among Vets was as high as that found in an academic veterinary hospital in our previous study. In contrast, only two blood donor dogs and one dog with liver disease (1.0%, 3/292) yielded MRSA. All MRSA-positive dogs were reared or treated in different VCs, in each of which at least one veterinary staff member carrying MRSA worked. Sequence types (ST) identified by multilocus sequence typing, spa types, and SCCmec types for canine MRSA isolates (ST5-spa t002-SCCmec II [from two dogs] or ST30-spa t021-SCCmec IV [from a dog]) were concordant with those from veterinary staff members in the same clinics as the MRSA-positive dogs, with which they had potentially had contact. Most MRSA isolates from veterinary staff were the same genotype (SCCmec type II and spa type t002) as a major hospital-acquired MRSA clone in Japan. The remaining MRSA was the same genotypes as domestic and foreign community-associated MRSA. Measures against MRSA infection should be provided in private VCs.

Epidemiological analysis of methicillin-resistant Staphylococcus aureus carriage among veterinary staff of companion animals in Japan.

Veterinary staff carrying methicillin-resistant Staphylococcus aureus(MRSA) can be a source of MRSA infection in animals. To identify risk factors of MRSA carriage among veterinary staff, MRSA carriage and epidemiological information (sex, career, contact with MRSA-identified animal patients and others) were analyzed from 96 veterinarians and 70 veterinary technicians working at 71 private veterinary clinics in Japan. Univariate analysis determined sex (percentage of MRSA carriage, male (29.2%) vs. female (10%); P=0.002) and career (veterinarians (22.9%) vs. veterinary technicians (10%); P=0.030) as risk factors. Multivariable analysis revealed that sex was independently associated with MRSA carriage (adjusted odds ratio, 3.717; 95% confidence interval, 1.555-8.889; P=0.003). Therefore, male veterinary staff had a higher risk of MRSA carriage than female staff.

[Familial type III hyperlipoproteinemia].

Familial type III hyperlipoproteinemia(HLP) is characterized by increased plasma triglyceride(TG) and plasma remnant lipoproteins(chyromicron remnants and VLDL remnants i.e. IDL). Remnants predispose affected subjects to premature or accelerated atherosclerosis. Clinical features of type III HLP with apo E2/2 genotype are examined in 26 Japanese patients. Mean levels of plasma TG, total cholesterol, LDL cholesterol and remnant cholesterol(RLP-C) were 374, 256, 74 and 49 mg/dL, respectively. High plasma RLP-C levels above 30 mg/dL and high plasma RLP-C/plasma TG ratio above 0.1 are very useful for diagnosis of type III HLP. Fifty-four point two percent of the patients had diabetes mellitus and 66.2 % of the patients had metabolic syndrome. Diabetes and obesity contribute to the occurrence of type III HLP with apo E2/2 genotype in Japan. Coronary heart disease(CHD) occurred in 41.7% of the patients. Type III HLP is strongly associated with atherosclerosis in Japan.

[Transient elevation of intraocular pressure in primary open-angle glaucoma patients after automated visual field examination in the winter].

PURPOSE: To evaluate retrospectively seasonal fluctuations of transient intraocular pressure (IOP) elevation after automated visual field examination in patients with primary open-angle glaucoma (POAG). PATIENTS AND METHODS: We reviewed 53 consecutive patients with POAG who visited Kaimeido Ophthalmic and Dental Clinic from January 2011 to March 2013, 21 men and 32 women aged 67.7 +/- 11.2 years. The patients were divided into 4 groups, spring, summer, autumn, and winter according to the month of automated visual field examination and both eyes of each patient were enrolled. IOP was measured immediately after automated visual field examination (vf IOP) and compared with the average IOP from the previous 3 months (pre IOP) and with the average IOP from the following 3 months (post IOP) in each season. IOP elevation rate was defined as (vf IOP- pre IOP)/pre IOP x 100% and calculated for each season (paired t test). Additionally, the correlation between mean deviation (MD) and IOP elevation rate was evaluated (single regression analysis). Exclusion criteria were patients who received cataract surgery during this study or had a history of any previous glaucoma surgery. The automated visual field test was performed with a Humphrey field analyzer and the Central 30-2 FASTPAC threshold program. RESULTS: The average vf IOP was 14.5 +/- 2.5 mmHg, higher than pre IOP 13.8 +/- 2.4 mmHg (p < 0.0001) and the post IOP 13.8 +/- 2.2 mmHg (p < 0.0001). IOP elevation rate in each season was 4.1 11.6% in spring (n = 22, p = 0.18), 0.1 +/- 9.9% in summer (n = 16, p = 1.0), 5.0 +/- 13.8% in autumn (n = 30. p = 0.11), 10.6 +/- 8.8% in winter (n = 38, p < 0.0001). The MD was not correlated with IOP elevation rate (p = 0.17). CONCLUSION: Patients with POAG show a transient IOP elevation after automated visual field examination, especially in the winter but not in the summer.

Apolipoprotein B-48 to triglyceride ratio is a novel and useful marker for detection of type III hyperlipidemia after antihyperlipidemic intervention.

AIM: Remnant lipoproteins are atherogenic and are accumulated in patients with type III hyperlipidemia (HL). Although type III HL is diagnosed by phenotyping apolipoprotein (apo) E, this procedure is time-consuming and inconvenient for routine clinical use. Clinical indices for screening type III HL in untreated HL patients have been proposed; however, in clinical settings, HL patients are promptly treated with lipid-lowering agents without diagnosing the underlying cause. We investigated whether existing clinical indices for screening type III HL as well as the apo B-48/triglyceride (TG) ratio, which was suggested to be related to the accumulation of small chylomicron (CM) remnants, are useful after the initiation of lipid-lowering therapies. METHODS: In 25 normolipidemic subjects and 191 treated HL patients (type I, n =6; IIa, 62; IIb, 66; III, 12; IV, 22; and V, 23) from Osaka University Hospital and related hospitals, fasting low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), TG, and apolipoproteins were measured and clinical indices were evaluated statistically. RESULTS: Apo B-48 levels were significantly higher in patients with type I, III, and V HL, and TG levels were significantly higher in patients with type I and V HL. The apo B-48/TG ratio was significantly higher only in patients with type III HL compared with other types of HL (p<0.001), and was statistically significant among the other clinical indices (AUC-ROC value, 0.895; cut-off value, 0.110). CONCLUSION: The apo B-48/TG ratio is a novel and useful marker for detecting type III HL even after the initiation of lipid-lowering interventions.

A novel complex deletion-insertion mutation mediated by Alu repetitive elements leads to lipoprotein lipase deficiency.

Lipoprotein lipase (LPL) deficiency is a rare autosomal recessive inherited disorder, characterized by marked hypertriglyceridemia, eruptive xanthoma, hepatosplenomegaly, recurrent attacks of pancreatitis, and markedly low or absent LPL activity in postheparin plasma. A majority of LPL deficient patients have been reported to have point mutations in the LPL gene; however, we find a complex deletion-insertion mutation by Alu elements, mobile retrotransposons, in a patient with LPL deficiency. This patient suffered from acute pancreatitis, showed chylomicronemia and lacked detectable LPL activity or mass in her postheparin plasma. Southern blot analysis and long-range PCR of the patient's DNA demonstrated a 2.2-kb deletion encompassing exon 2. Sequence analysis revealed (1) a 2.3-kb deletion between an AT-rich region adjacent to an Alu element in intron 1 and another Alu element in intron 2; (2) an insertion of approximately 150bp 5'-truncated Alu sequence with a poly (A) tail at the deletion point. The inserted sequence belongs to Alu Yb9, the youngest subfamily of Alu elements. The deletion occurred at the consensus cleavage site (3'-A|TTTT-5') without target site duplication. These findings indicated that Alu retrotransposition caused the complex deletion-insertion. The patient was homozygous for this complex mutation, which eliminates exon 2 and leads to LPL deficiency. To our knowledge, the patient is the first case with LPL deficiency due to a complex deletion-insertion mediated by Alu repetitive elements.

Plasma lipid levels and nutritional intake in childhood- and adolescence-onset young type 1 diabetic patients in Japan.

In recent years, the diet of the young Japanese has changed to westernized diet with high fat content. Childhood-onset type 1 diabetic patients have had good diet training since onset of the disease, but adolescence-onset type 1 diabetic patients have already established westernized diet habit at onset of the disease, which may not be easily improved. We hypothesized that a difference of the age at onset of the disease may affect nutritional status and plasma lipid levels in Japanese type 1 diabetic patients. Plasma lipid levels and nutritional intake were compared between childhood- and adolescence-onset young type 1 diabetic patients. Our research involved 9 childhood-onset type 1 diabetic patients (childhood group), 11 adolescence-onset type 1 diabetic patients (adolescent group), and 24 age-matched non-diabetic control subjects. There were no significant differences in age and body mass index (BMI), daily energy intake among the childhood group, the adolescent group, and the non-diabetic control group. There was no significant difference in HbA1c level between the childhood group and the adolescent group. The adolescent group had significantly higher plasma levels of total cholesterol, triglyceride, and low-density lipoprotein (LDL)-cholesterol than the childhood group (p<0.01, <0.05, and <0.001, respectively) or the control group (p<0.001, <0.001, and <00.001, respectively). The adolescent group had significantly lower plasma level of high-density lipoprotein (HDL)-cholesterol than the childhood group (p<0.05). The adolescent group had significantly higher percentage energy intake from fat (31.7%, p<0.001), higher saturated fatty acids intake (19.0g/day, p<0.01), and higher cholesterol intake (428mg/day, p<0.05), and significantly lower polyunsaturated fatty acids intake (13.4g/day, p<0.05) and lower fiber intake (9.5g/day, p<0.01) than the childhood group. It is concluded that young Japanese type 1 diabetic patients with onset of adolescence have lipid abnormalities, which may be mainly caused by westernized dietary habits.

Effect of apolipoprotein E4 allele on plasma LDL cholesterol response to diet therapy in type 2 diabetic patients.

OBJECTIVE: The aim of this study was to investigate the effect of apolipoprotein (apo)E4 allele on plasma LDL cholesterol response to calorie-restricted diet therapy in type 2 diabetic patients. RESEARCH DESIGN AND METHODS: Twenty-four diabetic patients with the apoE3/3 genotype and 11 diabetic patients with the apoE4/3 genotype were recruited. Participants were hospitalized for calorie-restricted diet therapy (25.0 kcal. kg body wt(-1). day(-1)) for 14 days. Body weight, fasting plasma glucose (FPG) levels, and plasma lipid levels on hospital days 1 and 14 were compared between the two apoE genotype groups. RESULTS: There were no significant differences in baseline FPG levels, HbA(1c) levels, BMI, and plasma levels of total cholesterol, triglyceride, and HDL cholesterol between the two apoE genotype groups, but baseline plasma levels of LDL cholesterol were significantly higher in the apoE4/3 group than in the apoE3/3 group. Body weight decreased slightly and FPG levels decreased significantly after diet therapy in both apoE genotype groups. In the apoE3/3 group, only plasma levels of triglyceride decreased significantly after diet therapy, whereas in the apoE4/3 group, plasma levels of triglyceride, total cholesterol, and LDL cholesterol decreased significantly after diet therapy. The decrease (percentage of change) in total cholesterol (-16.3 vs. -6.6%) and LDL cholesterol (-15.6 vs. -0.7%) after diet therapy was significantly greater in the apoE4/3 group than in the apoE3/3 group. CONCLUSIONS: Calorie-restricted diet therapy is more effective in reducing plasma LDL cholesterol in type 2 diabetic patients with the apoE4 allele.

Rescue of beta-cell exhaustion by diazoxide after the development of diabetes mellitus in rats with streptozotocin-induced diabetes.

In this study, we attempted to demonstrate the possibility of rescuing beta-cell exhaustion by chronic intervention with an ATP-sensitive K(+) channel opener, diazoxide, which reduces the stress of insulin secretion, using rats with streptozotocin-induced diabetes. Three groups of male Wistar rats: (i) controls (n = 7), (ii) streptozotocin (30 mg/kg i.v.)-induced diabetic rats (n = 10), and (iii) streptozotocin-induced diabetic rats treated with diazoxide 30 mg/kg for 6 weeks (n = 10), were studied. Intraperitoneal 2-g glucose tolerance testing was performed every 2 weeks, and pancreatic tissue was examined after 6 weeks of treatment with diazoxide. The insulin concentration in diabetic rats treated with diazoxide was significantly higher than in diabetic rats without diazoxide (6.6 +/- 1.6 vs. 2.4 +/- 1.0 ng/ml, P < 0.05). The islet size and its cell number were reduced in diabetic rats compared to those in normal control rats. In normal control rats, 88% of pancreatic islet cells were insulin-positive, while 50% or less were positive in diabetic rats. However, islet size and its cell size appeared to be well preserved by diazoxide treatment. The average mass of islets in diazoxide-treated rats was significantly larger than that in untreated control animals. In addition, the degree of immunostaining for insulin was obviously higher in rats treated with diazoxide than in rats without diazoxide. Pancreatic proinsulin mRNA was restored in rats treated with diazoxide. The present study demonstrated that diazoxide protected from further damage the pancreatic beta-cells both functionally and morphologically in streptozotocin-induced diabetic rats by suppression of excessive insulin secretion. Our results strongly suggest the possibility that chronic intervention with an ATP-sensitive K(+) channel opener prevents the progress of deranged beta-cell function even after the development of diabetes mellitus.

Remnant-like lipoprotein particles in type 2 diabetic patients with apolipoprotein E3/3 and apolipoprotein E2 genotypes.

Apolipoprotein (apo) E2 and diabetes mellitus are known to be associated with an accumulation of remnant lipoproteins in plasma. In this study, effects of type 2 diabetes mellitus and/or apo E2 genotypes on remnant-like lipoprotein particles (RLP) were assessed. Thirty-three subjects were divided into 6 groups: 7 apo E3/3 nondiabetic subjects, 6 apo E3/3 diabetic patients, 5 apo E3/2 nondiabetic subjects, 6 apo E3/2 diabetic patients, 5 apo E2/2 nondiabetic subjects, and 4 apo E2/2 diabetic patients. First, the effect of diabetes mellitus on RLP were estimated by comparing the apo E3/3 nondiabetic group with the apo E3/3 diabetic group. Plasma levels of RLP-cholesterol (chol) in the apo E3/3 diabetic group and the uptake of RLP from the apo E3/3 diabetic group by macrophages were significantly greater compared with the apo E3/3 nondiabetic group. Second, the effect of apo E2 on RLP was estimated in nondiabetic subjects. Apo E2/2 nondiabetic subjects had type III hyperlipoproteinemia (HLP). Plasma levels of RLP-chol in the apo E2/2 nondiabetic group and the uptake of RLP from the apo E2/2 nondiabetic group by macrophages were significantly greater compared with the apo E3/3 and apo E3/2 nondiabetic groups. Third, the effects of both apo E2 and diabetes on RLP were estimated. Plasma levels of RLP-chol in the apo E2 (E3/2 and E2/2) diabetic groups and the uptake of RLP from apo E2 (E3/2 and E2/2) diabetic groups by macrophages were significantly greater compared with apo E3/3 nondiabetic and diabetic groups or the apo E3/2 nondiabetic group. In diabetes, a gene dose effect of apo E2 on plasma levels of RLP-chol and uptake of RLP by macrophages was present (apo E3/3 < apo E3/2 < apo E2/2). The apo E2/2 diabetic group had type III HLP. Furthermore, uptake of RLP from the apo E2/2 diabetic group with type III HLP was significantly greater compared with the apo E2/2 nondiabetic group with type III HLP. In conclusion, type 2 diabetes was associated with increased RLP-chol in plasma and atherogenic RLP. In nondiabetes, apo E2/2 contributes to increased plasma RLP-chol and atherogenic RLP. In diabetes, additional effects of apo E2 to increase RLP-chol in plasma and to enhance the uptake of RLP by macrophages are present. RLP from apo E2/2 diabetes with type III HLP are more atherogenic than those from apo E2/2 nondiabetes with type III HLP.

Apolipoprotein E genetic polymorphism, remnant lipoproteins, and nephropathy in type 2 diabetic patients.

BACKGROUND: We previously showed that the apolipoprotein (apo) Eepsilon2 allele is associated with the progression of diabetic nephropathy. The aim of the present study is to further investigate the association between apo E genetic polymorphism, plasma lipid levels (particularly remnant lipoproteins), and diabetic nephropathy. SUBJECTS AND METHODS: One hundred fifty-eight patients with type 2 diabetes who had a duration of diabetes longer than 10 years were divided into the three apo E groups: apo E2 (n = 22), E3/3 (n = 102), and E4 (n = 34). Plasma levels of lipids and remnant lipoproteins were measured. The effect of apo E2 triglyceride (TG)-rich lipoproteins, including remnant lipoproteins, on the accumulation of cholesteryl esters by human mesangial cells (HMCs) was estimated by measuring the stimulation of radioactive carbon-labeled oleate incorporation into cholesteryl esters. RESULTS: The frequency of overt nephropathy was significantly greater in apo E2 patients with diabetes (59.1%) than apo E3/3 (34.3%) or apo E4 patients (8.8%), and the frequency of normoalbuminuria was significantly greater in apo E4 patients with diabetes (67.6%) than apo E3/3 (34.3%) or apo E2 patients (4.5%). Logistical regression analysis showed that odds ratios of apo E2 and apo E4 genotypes for the presence of overt nephropathy were 10.179 (P = 0.0349) and 0.129 (P = 0.0028), respectively. Plasma TG and remnant-like lipoprotein particle cholesterol levels were significantly greater in apo E2 patients and significantly lower in apo E4 patients than apo E3/3 patients. Apo E2 TG-rich lipoproteins stimulated the accumulation of cholesteryl esters by HMCs significantly more than apo E3/3 or apo E4 TG-rich lipoproteins. CONCLUSION: Apo E2 is a positive factor and apo E4 is a negative factor for diabetic nephropathy. Apo E2 TG-rich lipoproteins, including remnant lipoproteins, affected HMCs. Remnant lipoproteins may have an important role in the progression of diabetic nephropathy.