Genetic correlation between the pre-adult developmental period and locomotor activity rhythm in Drosophila melanogaster.

Biological clocks regulate various behavioural and physiological traits; slower circadian clocks are expected to slow down the development, suggesting a potential genetic correlation between the developmental period and circadian rhythm. However, a correlation between natural genetic variations in the developmental period and circadian rhythm has only been found in Bactrocera cucurbitae. The number of genetic factors that contribute to this genetic correlation is largely unclear. In this study, to examine whether natural genetic variations in the developmental period and circadian rhythm are correlated in Drosophila melanogaster, we performed an artificial disruptive selection on the developmental periods using wild-type strains and evaluated the circadian rhythms of the selected lines. To investigate whether multiple genetic factors mediate the genetic correlation, we reanalyzed previously published genome-wide deficiency screening data based on DrosDel isogenic deficiency strains and evaluated the effect of 438 genomic deficiencies on the developmental periods. We then randomly selected 32 genomic deficiencies with significant effects on the developmental periods and tested their effects on circadian rhythms. As a result, we found a significant response to selection for longer developmental periods and their correlated effects on circadian rhythms of the selected lines. We also found that 18 genomic regions had significant effects on the developmental periods and circadian rhythms, indicating their potential for mediating the genetic correlation between the developmental period and circadian rhythm. The novel findings of our study might lead to a better understanding of how this correlation is regulated genetically in broader taxonomic groups.

Inactivation of creatine kinase induced by quercetin with horseradish peroxidase and hydrogen peroxide. pro-oxidative and anti-oxidative actions of quercetin.

Pro-oxidative and anti-oxidative actions of quercetin were examined through inactivation of CK and inhibition of lipid peroxidation. Quercetin induced inactivation of creatine kinase (CK) during the interaction with horseradish peroxidase and hydrogen peroxide (HRP-H(2)O(2)). CK activity in heart homogenate was also reduced by quercetin with HRP-H(2)O(2). Flavonoids that have a catechol structure in the B ring, such as taxifolin, catechin and luteolin, also induced CK inactivation. These flavonoids strongly inhibited NADPH and ADP-Fe(3+)-dependent microsomal lipid peroxidation. These results suggest a close relationship between pro-oxidative and anti-oxidative actions of quercetin. Electron spin resonance (ESR) signals of the quercetin radical was emitted during the interaction of quercetin with HRP-H(2)O(2) in the presence of Zn(2+) as a stabilizer. Adding CK diminished the ESR signals of quercetin radicals, suggesting CK efficiently scavenged quercetin radicals. Sulfhydryl groups and tryptophan residues in CK decreased during the interaction of quercetin with HRP-H(2)O(2). The kinetic parameters of K(m) and V(max) for ADP and creatine phosphate changed rapidly, suggesting that the inactivation of CK was induced through conformational change of the enzyme. Glyceraldehyde-3-phosphate dehydrogenase had a higher sensitivity to quercetin with HRP-H(2)O(2) than CK. Quercetin radicals may mediate between pro-oxidative and anti-oxidative action.

A developmentally regulated and psychostimulant-inducible novel rat gene mrt1 encoding PDZ-PX proteins isolated in the neocortex.

Single or repeated exposure to psychostimulants such as amphetamines and cocaine after postnatal week 3 leads to an enduring enhancement in the psychotomimetic responses elicited by a subsequent challenge of a stimulant in rodents. This behavioral sensitization phenomenon has been considered to be the neural consequences of stimulant-induced alterations in gene expression in the brain after a critical period of postnatal development. Using a differential cloning technique, RNA arbitrarily primed PCR, we have now identified from the rat neocortex a novel and developmentally regulated methamphetamine (MAP)-inducible gene mrt1 (MAP responsive transcript 1). mrt1 encodes two major types of PDZ- and PX-domains containing proteins of approximately 62 kDa in size with different carboxy termini, Mrt1a and Mrt1b. The mrt1 mRNAs for Mrt1a, mrt1a, and for Mrt1b, mrt1b, are predominantly expressed in various brain regions and the testes, respectively. Acute MAP injection upregulated mrt1b expression in the neocortex after postnatal week 3 in a D1 receptor antagonist-sensitive manner without affecting mrt1a expression. This upregulation was mimicked by another stimulant, cocaine, whereas pentobarbital and D1 antagonist failed to change the mrt1b transcript levels. Moreover, repeated daily treatment of MAP, but not MAP plus D1 antagonist, for 5 days caused an augmentation of the basal expression of mrt1b 2 and 3 weeks after the drug discontinuation. These late-developing, cocaine-crossreactive, D1 antagonist-sensitive and long-term regulations of mrt1b by MAP are similar to the pharmacological profiles of stimulant-induced behavioral sensitization, and therefore may be associated with the initiation and/or maintenance of the long-term neuronal adaptation.

Pseudoankylosis of the mandible as a result of methyl methacrylate-induced inflammatory cicatricial contracture of the temporal muscle after cranioplasty.

Pseudoankylosis of the mandible after intracranial surgical procedure has been widely reported, and is usually caused by fibrosis of the temporal muscle as a result of injury during the operation. We present an unusual case of mandibular pseudoankylosis as a result of methyl methacrylate-induced aseptic inflammatory cicatricial contracture of the temporal muscle after cranioplasty.

Amatoxin poisoning from ingestion of Japanese Galerina mushrooms.

BACKGROUND: Although some Japanese Galerina species poisonings manifest as gastrointestinal symptoms followed by late-onset hepatorenal failure (phalloides syndrome), the toxin responsible for this has not been determined. CASE REPORT: We report a 6-year-old boy who developed characteristic cholera-like diarrhea and late-onset severe hepatic deterioration after eating mushrooms, later identified as a Galerina species, most likely Galerina fasciculata. A residual mushroom revealed alpha-amanitin. This account is the first known reported case of poisoning by Japanese Galerina species where an amatoxin was demonstrated to be responsible for the toxicity.

Forced swimming stimulates the expression of vasopressin and oxytocin in magnocellular neurons of the rat hypothalamic paraventricular nucleus.

Previous studies have shown that a 10-min forced swimming session triggers the release of both vasopressin and oxytocin into the extracellular fluid of the hypothalamic paraventricular (PVN) and supraoptic nuclei (SON) in rats. At the same time oxytocin, but not vasopressin, was released from the axon terminals into the blood. Here we combined forced swimming with in situ hybridization to investigate whether (i) the stressor-induced release of vasopressin and oxytocin within the PVN originates from parvo- or magnocellular neurons of the nucleus, and (ii) central release with or without concomitant peripheral secretion is followed by changes in the synthesis of vasopressin and/or oxytocin. Adult male Wistar rats were killed 2, 4 or 8 h after a 10-min forced swimming session and their brains processed for in situ hybridization using 35S-labelled oligonucleotide probes. As measured on photo-emulsion-coated slides, cellular vasopressin mRNA concentration increased in magnocellular PVN neurons 2 and 4 h after swimming (P < 0.05). Similarly, oxytocin mRNA concentration was significantly increased in magnocellular neurons of the PVN at 2 and 8 h (P < 0.05). We failed to observe significant effects on vasopressin and oxytocin mRNA levels in the parvocellular PVN and in the SON. Taken together with results from previous studies, our data suggest that magnocellular neurons are the predominant source of vasopressin and oxytocin released within PVN in response to forced swimming. Furthermore, in the case of vasopressin, central release in the absence of peripheral secretion is followed by increased mRNA levels, implying a refill of depleted somato-dendritic vasopressin stores. Within the SON, however, mRNA levels are poor indicators of the secretory activity of magnocellular neurons during stress.

Incidence of deep fascial space infection after surgical removal of the mandibular third molars.

Nine hundred and ninety-three patients who underwent surgical removal of the mandibular third molars with oral antibiotic prophylaxis were examined to determine the incidence of postoperative deep fascial space infection and its background factors. Postoperative deep fascial space infection was observed in 8 of the patients (0.8%; 4 males and 4 females), and submandibular spaces were involved in all infected patients. Only 1 of these 8 patients was an immune compromised host. Patients aged 30 years or more had a significantly higher incidence of deep fascial space infection than those aged under 30. Five patients had partial bony impactions and 3 had complete bony impactions. However, the incidence of infection according to the molar positions was not significantly different between partial bony impaction and complete bony impaction. The 8 patients had not had pericoronitis preoperatively. The clinical courses of all were favorable after antibiotics were administered intravenously. In conclusion, the incidence of deep fascial space infection after removal of the mandibular third molars was low, at 0.8%. However, it may be desirable to remove the molars, if applicable, at a younger age because of the higher incidence of infection in patients aged over 30. The results of this study also offer information that will be useful as a basis for obtaining informed consent from patients whose mandibular third molars are to be removed.

Specific characteristics and management strategies of cerebral artery aneurysms: report of eleven cases.

Owing to the deep location of the posterior cerebral artery (PCA) and its close relationship with the brainstem and surrounding vital structures, surgical treatment of aneurysms in this region is complex. This study was undertaken in an attempt to better delineate the surgical risks of PCA aneurysms. A retrospective analysis was undertaken in 11 patients with PCA aneurysm surgically treated between 1988 and 1996 at Shinshu University and its affiliated hospitals. Data regarding surgical strategy, surgical complications and outcomes were analysed. Seven aneurysms were saccular (including one mycotic) and the other four were fusiform, dissecting, thrombosed and an infundibular dilatation. The locations of the aneurysms were at the P1 segment in two patients, P1-P2 junction in two, P2 segment in six and P3 segment in one. Six saccular non-mycotic aneurysms were treated with neck clipping and the other five aneurysms were treated each with proximal occlusion of the parent artery, excision of the aneurysm or wrapping. All aneurysms were satisfactorily exposed except one large saccular aneurysm. Surgical outcomes were either good recovery or moderate disability in 10 patients, and severe disability in one patient with a large aneurysm due to temporal lobe contusion. In conclusion it is the responsibility of the surgeon dealing with rare PCA aneurysms to be aware of these specific characteristics and to appreciate which surgical technique is appropriate for each patient.

Phenylbutazone radicals inactivate creatine kinase.

Creatine kinase (CK) was used as a marker molecule to examine the side effect of damage to tissues by phenylbutazone (PB), an effective drug to treat rheumatic and arthritic diseases, with horseradish peroxidase and hydrogen peroxide (HRP-H(2)O2). PB inactivated CK during its interaction with HRP-H(2) O(2), and inactivated CK in rat heart homogenate. PB carbon-centered radicals were formed during the interaction of PB with HRP-H(2)O2. The CK efficiently reduced electron spin resonance signals of the PB carbon-centered radicals. The spin trap agent 2-methyl-2-nitrosopropane strongly prevented CK inactivation. These results show that CK was inactivated through interaction with PB carbon-centered radicals. Sulfhydryl groups and tryptophan residues in CK were lost during the interaction of PB with HRP-H(2)O2, suggesting that cysteine and tryptophan residues are oxidized by PB carbon-centered radicals. Other enzymes, including alcohol dehydrogenase, glyceraldehyde-3-phosphate dehydrogenase, but not lactate dehydrogenase, were also inactivated. Sulfhydryl enzymes seem to be sensitive to attack by PB carbon-centered radicals. Inhibition of SH enzymes may explain some of the deleterious effects induced by PB.

Inactivation of creatine kinase during the interaction of indomethacin with horseradish peroxidase and hydrogen peroxide: involvement of indomethacin radicals.

Creatine kinase (CK) was used as a marker molecule to examine the side effect of damage to tissues by indomethacin (IM), an effective drug to treat rheumatoid arthritis and gout, with horseradish peroxidase and hydrogen peroxide (HRP-H2O2). IM inactivated CK during its interaction with HRP-H2O2. Under aerobic conditions, inactivation of CK significantly decreased. CK in rat heart homogenate was also inactivated by IM with HRP-H2O2. When IM was incubated with HRP-H2O2, the maximum absorption of IM at 280 nm rapidly decreased and a new peak at 410 nm occurred with isosbestic points at 260 and 312 nm. In contrast, under anaerobic conditions, the spectral change of IM was almost absent, indicating IM was oxidized to the yellow substance by HRP-H2O2. Adding catalase strongly inhibited the production of yellow substance. Sodium azide also blocked the formation of yellow substance and the inactivation of CK. Electron spin resonance signals of IM carbon-centered radical were detected using 2-methyl-2-nitrosopropane during the interaction of IM with HRP-H2O2 under anaerobic conditions. Oxygen was consumed during the interaction of IM with HRP-H2O2. These results suggest that IM carbon-centered radicals may rapidly react with O2 to generate the peroxyl radicals. Sulfhydryl groups and tryptophane residues of CK decreased during the interaction of IM with HRP-H2O2. Other sulfhydryl enzymes, including alcohol dehydrogenase and glyceraldehyde-3-phosphate dehydrogenase, were also readily inactivated during the interaction with HRP-H2O2. Sulfhydryl enzymes seem to be very sensitive to IM activated by HRP-H2O2.

DNA strand break and 8-hydroxyguanine formation induced by 2-hydroxyestradiol dispersed in liposomes.

DNA damage induced by estrogens dispersed in liposomes was investigated. 2-Hydroxyestradiol (2HOE(2)) and 4-hydroxyestradiol (4HOE(2)), but not estrone, estradiol-17beta or estriol, caused strand break of plasmid DNA damage in the presence of ADP-Fe(3+). The catechol structure may be necessary for DNA damage. When DNA was incubated with 2HOE(2) for a long time (24 h), DNA damage was induced even at very low concentrations. Adding hydrogen peroxide markedly enhanced the sensitivity of DNA to the attack by 2HOE(2). Hydroxyl radical (HO.) scavengers strongly inhibited the 2HOE(2)-induced DNA damage, and EDTA partially inhibited DNA damage. However, 2HOE(2) caused 8-hydroxyguanine formation from calf thymus DNA only in the presence of EDTA-Fe(3+), but not ADP-Fe(3+). In addition, deoxyribose, which is a detective molecule of HO(.), was not degraded by 2HOE(2) in the presence of ADP-Fe(3+). Upon adding EDTA 2HOE(2) rapidly degraded deoxyribose. These results suggest that DNA strand break caused by 2HOE(2) in the presence of ADP-Fe(3+) was due to ferryl ion rather than HO(.), whereas 8-hydroxyguanine (8HOG) induced by 2HOE(2) in the presence of EDTA-Fe(3+) was due to HO(.).

Strategic embolisation for successful resection of a large cerebral arteriovenous malformation.

The risks accompanied by the treatment of cerebral arteriovenous malformation (AVM) are still cumulative despite recent progress in available treatment options. Pre-operative embolisation is one such option, however, it seldom makes the surgical resection difficult. The excessive embolised nidus makes the surgical resection difficult because it cannot be compressed during the resection surgery and embolised nidus as a 'glue ball' with marginal hypervascular territory is most difficult to remove. The aim of pre-operative embolisation for successful surgical resection is to put glue into the marginal part of the nidus so as to make a cleavage between the surrounding normal tissues. Remaining feedings via the dilatated leptomeningeal anastomoses from surrounding normal cortical arteries do not interfere with the resection and can be eliminated easily by coagulating the pia matter around the nidus. Strategic planning with regard to the systemic course of treatment, including the manner of resection, is important for effective pre-operative embolisation.

Iron-deficiency anemia associated with Helicobacter pylori gastritis.

BACKGROUND: Recent studies have suggested an association of Helicobacter pylori and iron-deficiency anemia (IDA). This is a report of six cases of IDA associated with H. pylori gastritis. METHODS: Six patients with IDA were studied (5 boys and 1 girl; mean age 13.6 years; range 13-15 years). Five had a medical history of long-standing IDA and of oral iron supplementation at outpatient clinics. The anemia recurred after the iron therapy had been discontinued. The sixth patient was admitted to the hospital with severe IDA. An extensive work-up was ordered that included technetium-99m (99mTc) scans for Meckel's diverticulum, total colonoscopy, and gastrointestinal endoscopy. After biopsy-based H. pylori test results were confirmed to be positive, anti-H. pylori therapy consisting of lansoprazole, clarithromycin, and metronidazole was administered for 2 weeks with no iron supplementation. The hematologic profile and iron status were assessed periodically after the end of the eradication regimen. RESULTS: Upper gastrointestinal endoscopy revealed a marked antral nodularity but no evidence of bleeding lesions in all the patients. Given the histology and the fact that rapid urease test results were positive, chronic active gastritis with H. pylori was diagnosed in all these cases. H. pylori was successfully eradicated in all the patients. There was no evidence of IDA in any of the follow-up examinations between 27 and 50 months after anti-H. pylori therapy. CONCLUSIONS: H. pylori infection may be involved in cases of IDA of unknown origin, and the eradication of H. pylori can be associated with the resolution of anemia.

DNA damage induced by catechol derivatives.

We investigated the effect of catechol derivatives, including dopa, dopamine, adrenaline and noradrenaline, on DNA damage and the mechanisms of DNA strand breakage and formation of 8-hydroxyguanine (8HOG). The catechol derivatives caused strand breakage of plasmid DNA in the presence of ADP-Fe(3+). The DNA damage was prevented by catalase, mannitol and dimethylsulfoxide, suggesting hydroxyl radical (HO..)-like species are involved in the strand breakage of DNA. Iron chelators, such as desferrioxamine and bathophenanthroline, and reduced glutathione also inhibited the DNA damage. Deoxyribose, a molecule that is used to detect HO,, was not degraded by dopa in the presence of ADP-Fe(3+). By adding EDTA, however, dopa induced the marked deoxyribose degradation in the presence of ADP-Fe(3+), indicating that EDTA may extract iron from ADP-Fe(3+) to catalyze HO. formation by dopa. Thus, EDTA was a good catalyst for HO.-generation, whereas it did not promote the strand breakage of DNA. However, calf thymus DNA base damage, which was detected as 8-HOG formation, was caused by dopa in the presence of EDTA-Fe(3+), but not in the presence of ADP-Fe(3+). The 8HOG formation was also inhibited by catalase and HO. scavengers, indicating that HO&z.rad; was involved in the base damage. These results suggest that DNA strand breakage is due to ferryl species rather than HO., and that 8HOG formation is due to HO. rather than ferryl species.

Antioxidative and prooxidative action of stilbene derivatives.

The effects of stilbene derivatives, including resveratrol, diethylstilboestrol and stilbene, as antioxidants or prooxidants were examined. Resveratrol and diethylstilboestrol, but not stilbene, strongly inhibited NADPH- and adenosine 5'-diphosphate (ADP)-Fe3+-dependent lipid peroxidation at the initial and propagation stages. In addition, phenolic stilbenes also inhibited ultraviolet light-induced lipid peroxidation. Resveratrol and diethylstilboestrol efficiently scavenged 2,2'-azobis-(2-amidinopropane)-dihydrochloride peroxyl radicals. However, 2,2'-diphenyl-p-picrylhydrazyl radicals were trapped only by resveratrol, but not by diethylstilboestrol. These results suggest that the inhibitory effect of phenolic stilbenes on lipid peroxidation was due to their scavenging ability of lipid peroxyl and/or carbon-cantered radicals. Resveratrol efficiently reduced ADP-Fe3+, but not EDTA-Fe3+. Stilbenes and diethylstilboestrol did not reduce either ADP-Fe3+ or EDTA-Fe3+. The strand breaks of DNA were stimulated during the interaction of resveratrol with ADP-Fe3+ in the presence of H2O2. These results suggest that phenolic stilbenes act as antioxidants of membrane lipids and that resveratrol has a prooxidative effect DNA damage during interaction with ADP-Fe3+ in the presence of H2O2.

Inactivation of creatine kinase by Adriamycin during interaction with horseradish peroxidase.

Oxidative damage of creatine kinase (CK) induced by Adriamycin((R)) (ADM) with peroxidase was investigated using horseradish peroxidase (HRP). ADM oxidatively inactivated CK during its interaction with HRP in the presence of H(2)O(2) (HRP-H(2)O(2)). The red color of ADM was lost during oxidation by HRP-H(2)O(2). Adding catalase stopped the color change of ADM induced by HRP-H(2)O(2), indicating that ADM was oxidized by HRP complex I or II. CK was inactivated readily, even when it was added to the reaction mixture containing colorless ADM. Some sulfhydryl groups of CK, which have an important role in its enzyme activity, were very sensitive to ADM activated by HRP-H(2)O(2), suggesting that inactivation of CK is due to oxidation of SH groups at the active center. Presumably, oxidative ADM quinone is involved dominantly in the inactivation of CK. Among the anthracycline drugs tested in this study, only ADM and epirubicin caused inactivation of CK and alcohol dehydrogenase and loss of the red color during oxidation by HRP-H(2)O(2).

Oxytocin induces preservation of social recognition in male rats by activating alpha-adrenoceptors of the olfactory bulb.

In this report, a series of four experiments was performed to evaluate the relationship between the olfactory bulb norepinephrine system and intra-olfactory bulb infusion of oxytocin in the preservation of social memory responses. The present data indicate that oxytocin exerts this preservation of social recognition through a specific, receptor-mediated mechanism within the olfactory bulb (experiment 1). The involvement of the olfactory bulb norepinephrine system is revealed by the demonstration that retrodialysis of oxytocin into the olfactory bulb increases norepinephrine release (experiment 4). Our data suggest that the increased output of olfactory bulb norepinephrine resulting from oxytocin appears to activate alpha-adrenoceptors to produce this preservation in recognition because infusions of clonidine into the olfactory bulb preserve recognition responses in a manner similar to that observed with oxytocin (experiment 2). In addition, a co-infusion of oxytocin with phentolamine abolishes recognition responses (experiment 3). Accordingly, this model affords the opportunity to study neuropeptide-catecholamine interactions, link these interactions with a specific behavioural outcome and identify a novel function/site of action for oxytocin in the male.

Effective production of amanitins by two-step cultivation of the basidiomycete, Galerina fasciculata GF-060.

Practical production of amanitins was attempted by fermentation using a basidiomycete, Galerina fasciculata GF-060. In liquid fermentation, intracellular alpha- and gamma-amanitins were the main products, while alpha- and beta-amanitins accumulated in solid cultured mycelia. The production of amanitins in liquid fermentation was strongly affected by the amount of the remaining carbon sources (particularly glucose and sucrose). In batch cultivation, the productivity of alpha-amanitin was 1.58 mg/l. To improve the productivity, replacement cultivation using glucose-free medium was attempted. As a result, the maximum production of alpha-amanitin reached 5.02 mg/l. These conditions (fermentation style and glucose starvation) are effective for the production of all the known types of amanitins.