Aberrant microRNA expression in the brains of neurodegenerative diseases: miR-29a decreased in Alzheimer disease brains targets neurone navigator 3.

AIMS: MicroRNAs (miRNAs) are small non-coding RNAs that regulate translational repression of target mRNAs. Accumulating evidence indicates that various miRNAs, expressed in a spatially and temporally controlled that manner in the brain plays a key role in neuronal development. However, at present, the pathological implication of aberrant miRNA expression in neurodegenerative events remains largely unknown. To identify miRNAs closely associated with neurodegeneration, we performed miRNA expression profiling of brain tissues of various neurodegenerative diseases. METHODS: We initially studied the frontal cortex derived from three amyotrophic lateral sclerosis patients by using a microarray of 723 human miRNAs. This was followed by enlargement of study population with quantitative RT-PCR analysis (n = 21). RESULTS: By microarray analysis, we identified up-regulation of miR-29a, miR-29b and miR-338-3p in amyotrophic lateral sclerosis brains, but due to a great interindividual variation, we could not validate these results by quantitative RT-PCR. However, we found significant down-regulation of miR-29a in Alzheimer disease (AD) brains. The database search on TargetScan, PicTar and miRBase Target identified neurone navigator 3 (NAV3), a regulator of axon guidance, as a principal target of miR-29a, and actually NAV3 mRNA levels were elevated in AD brains. MiR-29a-mediated down-regulation of NAV3 was verified by the luciferase reporter assay. By immunohistochemistry, NAV3 expression was most evidently enhanced in degenerating pyramidal neurones in the cerebral cortex of AD. CONCLUSIONS: These observations suggest the hypothesis that underexpression of miR-29a affects neurodegenerative processes by enhancing neuronal NAV3 expression in AD brains.

Epithelial overexpression of interleukin-32alpha in inflammatory bowel disease.

Interleukin (IL)-32 is a recently described proinflammatory cytokine, characterized by induction of nuclear factor (NF)-kappaB activation. We studied IL-32alpha expression in the inflamed mucosa of inflammatory bowel disease (IBD). We also investigated mechanisms regulating IL-32alpha expression. Tissue samples were obtained endoscopically or surgically from patients with ulcerative colitis (UC) (n = 10), Crohn's disease (CD) (n = 10), ischaemic colitis (n = 4) and normal colorectal tissues (n = 10). IL-32alpha expression was evaluated by standard immunohistochemical procedure. IL-32 mRNA expression was analysed by Northern blot. IL-32alpha was expressed weakly by colonic epithelial cells from normal individuals and subjects with ischaemic colitis. In the inflamed mucosa of IBD patients, epithelial IL-32alpha expression was increased markedly. In UC and CD patients, IL-32alpha expression was enhanced in affected mucosa compared to non-affected mucosa. In intestinal epithelial cell lines, expression of IL-32alpha mRNA and protein was enhanced by IL-1beta, interferon (IFN)-gamma and tumour necrosis factor (TNF)-alpha. A combination of TNF-alpha plus IFN-gamma exerted synergistic effects. IL-32alpha induction by IL-1beta and/or TNF-alpha was mediated by NF-kappaB activation. Epithelial IL-32alpha expression was increased in IBD patients, and in CD patients in particular. IL-32alpha might be involved in the pathophysiology of IBD as a proinflammatory cytokine and a mediator of innate immune response.

Refraction and Anterior Chamber Depth Change After Vitrectomy for Pseudophakia.

Purpose: To evaluate the association between the vitreous and the refractive error in pseudophakia.Methods: Vitrectomy was performed in 67 eyes of 61 patients who underwent cataract surgery. Vitrectomy was needed for epiretinal membrane in 30 eyes, macular edema in 22 eyes, macular hole in 3 eyes, lamelar macular hole in 2 eyes, macular hole in 3 eyes, lamelar macular hole in 2 eyes, vitreous opacity in 6 eyes, and vitreous hemorrhage in 4 eyes. Refraction was measured before the operation, and 1 month, 3 months, 6 months, and 12 months after operation. Anterior chamber depth was measured in 10 eyes before the operation, and 1 month, 3 months, and 6 months after operation. We evaluated the refractive error after vitrectomy in 49 eyes that had predicted refraction.Results: The mean value of refractive change was -0.3 D 1 month postoperatively, and then gradually became positive. The variation of refraction (6-month postoperative refraction minus preoperative refraction) was negatively correlated with preoperative refraction (P =.0052, R(2) = 0.146). If preoperative refraction was more myopic than -1.5 D, then refraction became positive. If preoperative refraction was not more myopic than -1.5 D, then refraction became negative.Conclusion: About 15% of postoperative refractive error may be associated with the vitreous, but further investigation is required.

[Refraction and anterior chamber depth change after vitrectomy for pseudophakia].

PURPOSE: To evaluate the association between the vitreous and the refractive error in pseudophakia. METHODS: Vitrectomy was performed in 67 eyes of 61 patients who underwent cataract surgery. Vitrectomy was needed for epiretinal membrane in 30 eyes, macular edema in 22 eyes, macular hole in 3 eyes, lamelar macular hole in 2 eyes, vitreous opacity in 6 eyes, and vitreous hemorrhage in 4 eyes. Refraction was measured before the operation, and 1 month, 3 months, 6 months, and 12 months after operation. Anterior chamber depth was measured in 10 eyes before the operation, and 1 month, 3 months, and 6 months after operation. We evaluated the refractive error after vitrectomy in 49 eyes that had predicted refraction. RESULTS: The mean value of refractive change was -0.3 D 1 month postoperatively, and then gradually became positive. The variation of refraction (6-month postoperative refraction minus preoperative refraction) was negatively correlated with preoperative refraction (p = 0.0052, R2 = 0.146). If preoperative refraction was more myopic than -1.5 D, then refraction became positive. If preoperative refraction was not more myopic than -1.5 D, then refraction became negative. CONCLUSION: About 15% of postoperative refractive error may be associated with the vitreous, but further investigation is required.

Vitreous opacification after neodymium:YAG posterior capsulotomy.

PURPOSE: To describe the clinical picture in eyes that developed vitreous opacification behind the intraocular lens (IOL) after neodymium:YAG (Nd:YAG) laser posterior capsulotomy and determine whether this type of opacification tends to occur in patients with diabetes. SETTING: Shinjo Eye Clinic, Miyazaki, Japan. METHODS: The clinical course in 728 eyes that had Nd:YAG posterior capsulotomy was reviewed. RESULTS: After Nd:YAG posterior capsulotomy, opacification developed in the vitreous in contact with the IOL in 9 eyes (1.2%). All occurred in diabetic patients, and the vitreous opacification developed within 1 month after the capsulotomy. A vitrectomy was performed in 8 eyes and in 1, the opacification spontaneously absorbed. Vitreous opacification occurred in 8.9% of 101 eyes of diabetic patients, and the prevalence in diabetic eyes was significantly higher than in nondiabetic eyes (P < .0001). Nine of the diabetic eyes were opaque and 92 nonopaque. In the opaque eyes, the prevalence of panretinal photocoagulation was higher than in the nonopaque eyes (P = .013), and hemoglobin Alc (P = .030) was higher; the interval between cataract surgery and Nd:YAG capsulotomy was shorter (P = .047) and the final visual acuity, lower (P = .045). CONCLUSION: The prevalence of vitreous opacification after Nd:YAG laser posterior capsulotomy was significantly higher in diabetic than in nondiabetic eyes. Viterectomy was effective for this type of opacification.

Change in postoperative refractive error when vitrectomy is added to intraocular lens implantation.

PURPOSE: To compare the actual and expected refractive errors after intraocular lens (IOL) implantation alone with those after IOL implantation with simultaneous vitrectomy. SETTING: Shinjo Eye Clinic, Miyazaki, Japan. METHOD: One hundred thirty-six eyes had cataract extraction and implantation of a single-piece IOL using a frown incision, continuous annular anterior capsule tear, phacoemulsification, and intracapsular lens fixation. Thirty-six eyes also had vitrectomy. RESULTS: Mean postoperative refractive error was 0.55 diopter (D) +/- 1.34 (SD) in eyes having no vitrectomy and 0.04 +/- 1.24 D in those having vitrectomy. The difference between groups was statistically significant (P = .047; t-test). CONCLUSION: The refraction after simultaneous IOL implantation and vitrectomy shifted toward myopia by a mean of 0.50 D compared with that after IOL implantation alone.

Myofibrils and Meat Texture of Broilers Fed on a Diet of Tochu Leaf Powder.

Feeding of Tochu leaf to broiler was done to discover the role of myofibrilar protein and the effects of Tochu on meat toughness. The quality and quantity of myofibrillar protein was the same between Tochu-fed muscle and the controls. The improvement of meat quality should be concluded to be due to collagen as reported before and not due to the myofibrillar proteins.

Analyses of metal ions in spider venoms in relation to insecticidal activity of clavamine.

To elucidate insecticidal activity of spider toxins, metal ions in venoms and in the bodies were determined by thin layer chromatography, spark source mass spectrometry, ion chromatography, inductively coupled plasma emission spectrometry and atomic absorption spectrometry. Two kinds of spiders were used, Nephila clavata and Nephila maculata. Metals from their venom glands were extracted with hydrochloric acid and the metal concentrations were almost the same in the two species. Many kinds of metals, Fe, Zn, Pb, Cu, Ca, Mg, Na, P and S were found at higher levels in the venoms at concentrations higher than in the bodies. The contents of metal ions were low in the dragonfly and the cicada which are considered to be preys. Clavamine, the main insecticidal component in N. clavata, was effective on larvae of a mosquito with Ca2+, Fe3+ or Pb2+, but ineffective with Mg2+, Zn2+, Fe2+ or Cu2+. It is suggested that the metal chelates play an important role in the intoxication and detoxication of the spider toxins.

Molecular analysis of genetic mutation in electrophoretic variant of human lactate dehydrogenase-A(M) subunit.

An electrophoretic variant of lactate dehydrogenase-A (M) subunit was discovered in a patient with multiple myeloma. DNA analysis of the variant allele revealed a nucleotide substitution (transition) of C to T at codon 314 (CGT-TGT), and this mutation resulted in the replacement of an arginine by a cysteine (R314C). This amino acid replacement affects the net charge of the subunit and makes the LDH-A variant have a faster electrophoretic mobility. The responsible missense mutation created a new restriction site, AGGCCT, which can be simply detected by endonuclease AatI digestion. In addition, four synonymous substitutions with no amino-acid replacements were found at codons 51, 119, 163 and 175 in the LDH-A gene from the patient.

Inputs from the nucleus of the solitary tract to subfornical organ neurons projecting to the paraventricular nucleus in the rat.

Extracellular single-unit activity was recorded from 26 subfornical organ (SFO) neurons antidromically identified as projecting to the hypothalamic paraventricular nucleus (PVN) in urethane-anesthetized male rats. Of these identified SFO neurons, 10 displayed an inhibitory response and 2 exhibited an excitatory response in neuronal excitability following electrical stimulation of the nucleus of the solitary tract (NTS), while the remaining neurons were unresponsive. Eight out of 10 SFO neurons that demonstrated the inhibitory response to NTS stimulation also displayed a reduction in ongoing activity in response to activation of peripheral baroreceptors, achieved by rising arterial blood pressure with an intravenous administration of the alpha-agonist metaraminol, whereas the remaining responsive neurons to NTS stimulation were ineffective. These results show that a part of SFO neurons projecting to the PVN may receive inhibitory inputs from the peripheral baroreceptors through NTS neurons.

Formation of 8-hydroxydeoxyguanosine in DNA treated with fecapentaene-12 and -14.

Fecapentaene-12 and -14, direct-acting mutagens in human feces, were found to hydroxylate the C-8 position of guanine residues in DNA in vitro. Fecapentaene-12 or -14 was incubated with 0.5 mg of calf thymus DNA in 1 ml of reaction mixture at pH 7.4 for 2 h at 37 degrees C in the dark, and then 8-hydroxydeoxyguanosine (8-OH-dG) was analyzed. In these conditions 8-OH-dG was formed dose-dependently at levels of 1.1-4.6 residues/10(4) dG with concentrations of 0.5-3.0 mM of fecapentaene-12. Similar results were obtained with fecapentaene-14. The amount of 8-OH-dG in untreated DNA was 0.2-0.3 residue/10(4) dG.

Formation of a mutagen, 2-amino-1-methyl-6-phenylimidazo[4,5-b]-pyridine (PhIP) in cooked beef, by heating a mixture containing creatinine, phenylalanine and glucose.

When a mixture of creatinine, phenylalanine and glucose in diethylene glycol-water solution was heated for 2 h at 128 degrees C, a mutagen, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), was found to be produced, identified by high-performance liquid chromatography, and UV-visible and mass spectrometries. The yield of PhIP was 3.6 nmoles/mmole original creatinine. PhIP was originally isolated from cooked beef, and this study shows creatinine, phenylalanine and glucose to be the probable precursors of PhIP in beef.

[Antihypertensive action of dl-alpha-tocopherol 5-n-butyl-2-pyridinecarboxylate (TF-80)].

Antihypertensive action of TF-80, an alpha-tocopheryl ester of fusaric acid (FA), was studied in normotensive and hypertensive rats. Oral doses of TF-80 up to 100 mg/kg did not affect systolic blood pressure in Wistar rats, whereas 50 and 100 mg/kg doses produced a significant reduction in systolic blood pressure in both spontaneously hypertensive rats (SHR) and DOCA-salt hypertensive rats (DSR) without significant changes in heart rate. Although maximal reduction in systolic blood pressure at single oral dosing of TF-80 (50, 100 mg/kg) and FA (15, 30 mg/kg) was approximately the same extent, respectively, the antihypertensive action of TF-80 was longer-lasting than that of FA. TF-80 at a chronic administration of 1 to 100 mg/kg/day for 6-9 weeks was preventive to the generation of hypertension in young SHR and DSR, but FA did not prevent the generation of hypertension. Hypotensive doses of TF-80 did not interfere in the rise in blood pressure produced by vasoactive substances such as norepinephrine, prostaglandin F2 alpha and angiotensin II in SHR, and it also failed to inhibit plasma renin. TF-80 was also found not to have a diuretic effect. These results show that TF-80 has a mild and long-lasting antihypertensive action that is different from FA. However, the antihypertensive mechanism of TF-80 remains to be solved.

[Study on the concentration of FT-207 and 5-FU in serum, lymph nodes and tissues after administration of FT-207 suppositories].

In 21 cancer patients receiving administration of FT-207 suppositories, we studied the concentration of FT-207 and 5-FU in the blood before surgery, and in the blood and tissues-tumor, normal and lymph node-sampled during surgery. These 21 cases were comprised of 12 cases of stomach cancer, 1 case of lymphosarcoma, 4 cases of rectal cancer, 2 cases of sigmoid cancer and 2 cases of breast cancer. Two hours after administration, the concentration of 5-FU reached its highest level and continued to maintain a level at which it could be expected to bring about clinical results even after four hours. The concentration of 5-FU in tumors and lymph nodes was high, especially in tumors compared with normal tissue (p less than 0.01). No difference could be found in the concentrations of FT-207 and 5-FU in serum and tissues from each 12 cases of stomach cancer, not only between of the early cancer and advanced cancer but also between patients more than 60 years old and those under 60 years old. No side-effects occurred in any case during suppository administration.