The expression pattern of PKCtheta in satellite cells of normal and regenerating muscle in the rat.

Protein kinase C (PKC) is a key enzyme in regulating a variety of cellular functions. PKCtheta is the most abundant PKC isoform expressed in skeletal muscle. However, the functional role of PKCtheta linked to muscle regeneration has not yet been identified. Using reverse transcription (RT)-PCR and immunofluorescence analysis, we investigated the expression patterns of PKCtheta in normal and regenerating tibialis anterior (TA) muscles in the rat. The amount of PKCtheta mRNA in the muscle increased from the 4th to 6th post-surgical day. Immunofluorescence revealed PKCtheta protein in quiescent satellite cells identified by c-Met. PKCtheta immunoreactivity was not observed in many proliferating satellite cells by labeling with BrdU in the regenerating muscle. At 4, 6 and 10 days postsurgery, PKCtheta immunoreactivity was observed in half the differentiating satellite cells labeling with myogenin. After 4 and 6 days, the localization of PKCtheta coincided with those of Pax7 and TGF-beta. Thus, PKCtheta may play an important role in inhibiting differentiation and maintaining the quiescent satellite cells in muscle regeneration.

Age-related reductions in expression of serum response factor and myocardin-related transcription factor A in mouse skeletal muscles.

The molecular signaling pathways linking the atrophy of skeletal muscle during aging have not been identified. Using reverse transcription (RT)-PCR, Western blotting, and immunofluorescence microscopy, we investigated whether the amounts of RhoA, RhoGDI, SRF, MRTF-A, and MyoD in the triceps brachii and quadriceps muscles change with aging in mice. Young adult (3 mo) and aged (24 mo) C57BL/6J mice were used. Senescent mice possessed many fibers with central nuclei in the quadriceps muscle. Western blotting using a homogenate of whole muscle or the cytosolic fraction clearly showed that the amount of SRF protein was significantly decreased in the aged skeletal muscles. Immunofluorescence labeling indicated more SRF-positive muscle fibers in young mice. Both young and old mice possessed SRF immunoreactivity in some satellite cells expressing Pax7. MRTF-A and STARS mRNA levels significantly declined with aging in the triceps brachii and quadriceps muscles. The amount of MRTF-A protein was markedly reduced in the nuclear fraction of aged muscle of mice. The amounts of RhoA and RhoGDI in the crude homogenate or the cytosolic and membrane fractions were greater in the aged muscle. Senescent mice possessed significantly higher levels of MyoD protein in the cytosol and nucleus. Decreased SRF and MRTF expression may induce the atrophy of skeletal muscle with aging.

Cyclosporin A modulates cellular localization of MEF2C protein and blocks fiber hypertrophy in the overloaded soleus muscle of mice.

The molecular signaling pathway linked to hypertrophy of the anti-gravity/postural soleus muscle after mechanical overloading has not been identified. Using reverse transcription-polymerase chain reaction (RT-PCR), Western blot, and immunohistochemical analyses, we investigated whether the amounts of myocyte enhancer factor (MEF)2C, MEF2D, and myogenin change in the mechanically overloaded soleus muscle after treatment with the calcineurin inhibitor cyclosporine A (CsA). Adult male ICR mice were subjected to a surgical ablation of the gastrocnemius muscle and treated with either CsA (25 mg/kg) or vehicle, once daily. They were killed at 2, 4, 7, 10, and 14 days post-injury. Mechanical overloading resulted in a significant increase in the wet weight and the cross-sectional area of slow and fast fibers of the soleus muscle in placebo-treated mice but not CsA-treated mice. RT-PCR analysis did not show a marked difference in MEF2C and MEF2D mRNA levels in the overloaded soleus muscle in placebo- or CsA-administered mice. After 2 days of mechanical overloading, we observed co-localization of MEF2C and myogenin in several mononuclear cells under both conditions. These MEF2C-positive mononuclear cells also possessed immunoreactivity for c-Met, a satellite cell marker. At 4 days, mechanical overloading induced marked expression of MEF2C but not MEF2D in the subsarcolemmal region in a group of myotubes and/or myofibers. Such a MEF2C-positive region emerged less often in the hypertrophied soleus muscle subjected to the treatment with CsA. At 7 days, we observed many mononuclear cells possessing both MEF2C and myogenin protein in mice treated with CsA, but not the placebo. Our results demonstrated that CsA treatment modulates the amount and cellular localization of MEF2C protein. The modulation of MEF2C by CsA treatment may inhibit the hypertrophic process in the soleus muscle after mechanical overloading.

Role of protein kinase Cbeta in rhythmic contractions of human pregnant myometrium.

To assess the role of protein kinase Cbeta (PKCbeta) in human myometrial contractions during pregnancy, we evaluated the effect of a PKCbeta inhibitor (LY333531) on the pregnant and nonpregnant myometrial contractions and compared the level of PKCbeta in the pregnant myometrium with that in the nonpregnant myometrium. The effects of LY333531 on the myometrial contractions were examined by measuring contractile activity (frequency and amplitude). PKCbeta in human myometrium was assessed at mRNA level using real-time PCR method. The characteristics of contractile activity were different between the pregnant and the nonpregnant myometrium. The amplitude of rhythmic contractions in the preterm and term myometrium was increased 2- to 2.5-fold when compared with that in the nonpregnant myometrium, but the frequency of rhythmic contractions was decreased by about half. LY333531 (10(-6) M) reduced the increased amplitude in the preterm and term myometrium by about 50%, and the inhibitory effects of LY333531 in the pregnant myometrium were significantly greater than that in the nonpregnant myometrium (about 50 vs 25%). However, the frequency in the pregnant and nonpregnant myometrium was not influenced by LY333531. Real-time PCR revealed a significant, five- to sevenfold increase in the expression of PKCbeta mRNA in the preterm and term myometrium when compared with the nonpregnant myometrium. These findings suggest that the increased amplitude of human myometrial contractions during pregnancy is related to the increased level of PKCbeta. A PKCbeta inhibitor may reduce preterm uterine contractions and prevent preterm delivery.

Role of tyrosine kinase-dependent phosphorylation of NR2B subunit-containing NMDA receptor in morphine reward.

We previously demonstrated that the morphine-induced rewarding effect was dramatically suppressed by cotreatment with an NR2B subunit-containing N-methyl-D-aspartate (NMDA) receptor antagonist ifenprodil. Therefore we propose here that the NR2B subunit-containing NMDA receptor may be involved in the rewarding effect of morphine. A growing body of evidence indicates that tyrosine kinases, such as Src family kinases, increase the phosphorylation of the intracellular C-terminal tyrosine residues on the NR2B subunit and potentiate NMDA receptor function. The following review provides our recent findings regarding the role of tyrosine kinase-dependent phosphorylation of the NR2B subunit-containing NMDA receptor in the development of psychological dependence on morphine.

Increased expression of neuregulin-1 in differentiating muscle satellite cells and in motoneurons during muscle regeneration.

Neuregulins belong to a family of multipotent growth-promoting proteins, and have been shown to have a crucial role in accumulating acetylcholine receptor at neuromuscular junctions. A functional role of neuregulins in muscle regeneration has not yet been identified. Using reverse transcription (RT)-PCR, Western blot and immunofluorescence analysis following bupivacaine injection into rat muscle, we investigated the expression pattern of neuregulin-1 (NRG-1) in normal and regenerating tibialis anterior (TA) muscle. In addition, we examined changes in NRG-1 expression in the spinal cord following muscle damage. Western blotting showed that muscle NRG-1 protein decreased soon after the damage, and increased gradually after the 4th day following the damage. The amount of NRG-1 mRNA in the muscle increased from the 2nd to 6th post-surgical day. The amount of NRG-1 protein, but not mRNA, increased gradually in the spinal cord after muscle damage. Immunofluorescence revealed NRG-1 protein in some quiescent satellite cells identified by c-Met. After 6 and 10 days, clear co-localization between NRG-1 and myogenin was noted in differentiating satellite cells. Thus, NRG-1 may play an important role in the differentiation of satellite cells in muscle regeneration, while increased NRG-1 expression in motoneurons may enhance the remodeling of partially damaged axons.

Overexpression of the chimeric gene of the floral regulator CONSTANS and the EAR motif repressor causes late flowering in Arabidopsis.

The transcription factor CONSTANS (CO) plays a central role in the photoperiod pathway by integrating the circadian clock and light signals into a control for flowering time. CO induces flowering locus T (FT) and suppressor of overexpression of CO 1 (SOC1) expression, and thereby promotes flowering. The ethylene-responsive element-binding factor associated amphiphilic repression (EAR) motif was used to construct a CONSTANS-EAR motif repressor gene (CO-Rep), which was overexpressed in Arabidopsis under the control of the Cauliflower mosaic virus 35S promoter in order to test its potential for flowering time regulation under inductive long day conditions. Morphological abnormalities in the root and cotyledon formation, and dwarfness were frequently seen in the transgenic plants, suggesting that the proper timing, location, and/or level of CO-Rep expression are important for its application. In morphologically normal CO-Rep plants, both bolting and flowering times under inductive long day conditions were twofold greater than in controls. As a result of the delay in flowering, rosette leaf number at bolting, and rosette and cauline leaf number at flowering increased significantly in CO-Rep plants. RT-PCR analysis demonstrated that FT expression was greatly reduced in the CO-Rep plants, while endogenous CO and SOC1 expression levels were not markedly affected. Conservation of CO among a diverse range of plant species, and its involvement in a variety of photoperiodic responses including flowering, suggests a high potential for use of CO-Rep to manipulate such responses in an agronomically desirable manner.

Regulation of neurotrophin-3 gene transcription by Sp3 and Sp4 in neurons.

Neurotrophin-3 (NT-3), a neurotrophin member, plays crucial roles in neuronal development, function and plasticity. Previous studies have demonstrated that NT-3 gene transcription is driven by alternative promoters A and B, located upstream of exons 1A (EIA) and 1B (EIB), respectively. However, the transcription factors and DNA elements that drive NT-3 gene transcription remain to be identified. Here, we analysed the promoter region of the NT-3 gene and found that an NT-3 transcript containing EIB is predominantly expressed in cortical neurons which preferentially utilize promoter B, and two tandemly repeated GC-boxes, located between -100 and -60 base pairs within promoter B, are required for the transcription. Electrophoretic mobility shift and chromatin immunoprecipitation assays revealed that both specificity protein (Sp)3 and Sp4 were able to bind to the Sp1 binding sequences within the GC boxes. Expression of dominant-negative Sp3 and Sp4 small interfering RNA in cortical neurons reduced the activity of the NT-3 gene promoter. Over-expression of Sp1 family members, especially Sp4, resulted in an increase of the NT-3 gene promoter. These findings indicate that the NT-3 gene is a target gene for Sp4 that is abundantly expressed in the brain.

Cigarette smoke extract enhances oxytocin-induced rhythmic contractions of rat and human preterm myometrium.

Although smoking during pregnancy is a major risk factor for preterm delivery, the underlying mechanism by which smoking stimulates uterine contractions is still poorly understood. In the present study, we tried to clarify the effects of smoking on myometrial contractility induced by oxytocin (OT) using cigarette smoke extract (CSE). Myometrial strips, which were taken from the rat on day 16 of pregnancy, and from human preterm and term delivery groups, were incubated overnight with several doses of CSE at 37 degrees C under non-hormonal conditions. The uterine contractile sensitivity and activity (force and frequency) upon exposure to OT were investigated. Furthermore, the expression levels of oxytocin receptor (OTR) mRNA in the myometrial strips were investigated by real-time PCR. Contractile sensitivity to OT in the rat CSE (10(-7) pieces/ml) group was found to be significantly higher than in the control group (P < 0.05). Contractile activity did not differ between the CSE and control groups. The expression levels of rat OTR mRNA in the CSE (10(-7) pieces/ml) group were significantly higher than in the control group (P < 0.01). Similarly, in preterm myometrial strips, the expression levels of human OTR mRNA in the CSE (10(-7) pieces/ml) group were significantly higher than in the control group (P < 0.05). These findings suggest that CSE directly increases the contractile sensitivity of preterm myometrium in response to OT by upregulating the expression of OTR mRNA and thereby increases the risk of preterm delivery in women, who smoke during pregnancy.

Normal distribution of presenilin-1 and nicastrin in skeletal muscle and the differential responses of these proteins after denervation.

Presenilin-1 and nicastrin, two components of gamma-secretase associated with Alzheimer's disease plaques, are present in the synapses of the brain and in various peripheral organs, including skeletal muscle. In the present study, we examined the expression pattern of presenilin-1 and nicastrin in normal and denervated hindlimb muscles of the rat. Using immunohistochemical approaches, we found that presenilin-1 and AChRalpha was co-localized at the neuromuscular junction in the normal skeletal muscles of rats. The immunoreactivities of both presenilin-1 and nicastrin were also observed at the sarcolemma of muscle fibers. We discovered that presenilin-1 mRNA and its protein are upregulated after denervation of the soleus and tibialis anterior muscles. Furthermore, clear co-localization between presenilin-1 and DAPI, but not nicastrin, was noted in several myonuclei in the denervated muscles. We recognized a few fibers possessing both ubiquitin and presenilin-1 protein in the cytosol. The amount of presenilin-1 in the nucleus and membrane fraction was more abundantly expressed in the denervated muscle fibers. In contrast, no significant difference in the nicastrin protein level was observed between normal and denervated muscle fibers. These data suggest that enhanced presenilin-1 protein may play a role in the degeneration and regeneration of skeletal muscle.

Acupuncture stimulates the release of serotonin, but not dopamine, in the rat nucleus accumbens.

Acupuncture has been introduced as one of the available therapies widely used in alternative medicine, but it has not achieved widespread acceptance with scientific evidence. Furthermore there are still many unanswered questions about the basic mechanisms of acupuncture. To investigate the neuropharmacological mechanisms of oriental acupuncture, we studied the acupuncture-induced changes of in vivo monoamine release in the rat brain. A microdialysis guide cannula was implanted into the nucleus accumbens (ACC), which plays an important role in the brain reward system. Acupuncture treatment at the unilateral or bilateral Shenshu (bladder urinary channel 23) acupoints, located on the both sides of the spinous processes on the lower back, was carried out for 60 min in freely moving rats, and the dopamine (DA) and serotonin (5-HT) contents of the microdialysates in the ACC were measured simultaneously. In rats subjected to acupuncture at bilateral Shenshu acupoints, increases of 5-HT release in the ACC were observed at 20 min of acupuncture treatment and continued until 40 min after acupuncture was ended. Acupuncture at a unilateral Shenshu acupoint increased the release of 5-HT at 20 min compared with that in the sham-control group. Five-HT release returned to the baseline level at 120 min. The effects of acupuncture at bilateral Shenshu acupoints on the release of 5-HT in the ACC were greater than that of unilateral acupuncture treatment. In contrast, DA release in the ACC was not changed following acupuncture treatment. Effective acupuncture increased and prolonged the activity of serotonergic neurons in the reward system pathway of the brain. This suggests that oriental acupuncture therapy may be effective for the treatment of emotional disorders, drug abuse and alcoholism.

Correlation p53 expression and human papilloma virus deoxyribonucleic acid with clinical outcome in early uterine cervical carcinoma.

BACKGROUND: In the present study we assessed whether expression of p53 protein or HPV DNA correlates with recurrence as well as several known prognostic factors in uterine cervical carcinoma. METHODS: Forty-nine patients with FIGO stage IA-IIB who underwent hysterectomy between 1998 and 2002 were retrospectively studied. All 49 cancer tissue samples were used for immunohistochemical study. Twenty-five of 49 cases were also examined by PCR-RFLP for detection and typing of HPV DNA. RESULTS: Twenty of 49 (40.8%) specimens demonstrated nuclear staining for p53. A significant association between p53 overexpression and age, hormonal status, FIGO stage, or recurrence was observed (p=0.02, 0.01, 0.03, 0.01). However, no significant association was found between p53 overexpression and lymph node metastases, parametrium involvement, or risk of death (p=0.18, 0.06, 0.14). Nineteen of 25 (76%) were HPV DNA-positive and 6 (24%) were negative. DISCUSSION: There was no relation between HPV DNA positivity and age, FIGO stage, lymph node metastases, parametrium involvement, recurrence, or risk of death. CONCLUSION: p53 overexpression is associated with age, hormonal status, FIGO stage, and recurrence in uterine cervical carcinoma.

Expression of the endothelial cell differentiation gene 7 (EDG-7), a lysophosphatidic acid receptor, in ovarian tumor.

AIM: Lysophosphatidic acid (LPA) has received attention as a mitogen because the physiologically active lipid stimulates ovarian cancer cell growth by interacting with specific receptors, the endothelial cell differentiation gene (EDG) family. In the present study, we have investigated the expression of EDG-7 mRNA, part of the EDG family, in both human ovarian cancers and established human ovarian cancer cell lines. METHODS: RNA was extracted from six ovarian cancer cell lines and multiple cancerous and normal ovarian tissues. The expression of EDG-7 mRNA was measured using reverse transcription-polymerase chain reaction and northern blotting, using reduced glyceraldehyde-phosphate dehydrogenase and S26 as internal controls. RESULTS: Of the cell lines tested, EDG-7 mRNA was expressed most intensely in CRL-11731 and CRL-1572 and at a lesser but still substantial level in CRL-11732. The expression of EDG-7 mRNA was limited in MCAS, CRL-11730 and TYKnu. In the ovarian cancer tissues, EDG-7 mRNA was expressed most highly in endometrioid adenocarcinoma and serous cystadenocarcinoma. The expression of EDG-7 mRNA was limited in clear cell adenocarcinoma and undetectable in mucinous cystadenocarcinoma. CONCLUSIONS: The intense EDG-7 expression in ovarian cancers suggests that the relation between LPA and EDG-7 (an LPA receptor) is involved in cancer cell growth and proliferation in some histologic subtypes of ovarian cancer.

Cyclosporin A treatment upregulates Id1 and Smad3 expression and delays skeletal muscle regeneration.

The molecular signaling pathway linked to muscle regeneration has not yet been identified. Previously, we demonstrated that mice treated with cyclosporin A (CsA), a calcineurin inhibitor, failed to regenerate normally after muscle damage. Using reverse transcription (RT)-PCR, Western blot and immunohistochemical analysis, we investigated whether the amounts of nuclear factor of activated T cells (NFAT), myocyte-enhancer factor 2 (MEF2), the MyoD family, Id-1, and Smad3 change in the regenerating muscle after CsA treatment. Adult male ICR mice were subjected to a bupivacaine injection into the tibialis anterior muscle, and were treated with either CsA (25 mg/kg) or vehicle once daily. They were killed at 1, 2, 4, 6, 9 and 14 days post injury. RT-PCR analysis did not show a significant difference in MEF2s, MyoD and myogenin mRNA levels in the regenerating muscle in either placebo- and CsA-administered mice. In contrast, a significant increase in MRF4 mRNA was seen in CsA-administered mice compared to the placebo-treated mice at 4 and 9 days post surgery. In CsA-treated mice, the level of Id1 mRNA was elevated at day 9 relative to the placebo-treated mice. After 6 days, the CsA-treated mice possessed more abundant proliferating cell nuclear antigen (PCNA) and cyclin D1 protein in many satellite cells and/or myoblast-like cells in the regenerating muscle. The amount of myostatin, TGF-beta2 and Smad3 mRNA and proteins was increased more markedly in the mice treated with CsA. After 9 days, many satellite cells and/or myoblasts showed apparent co-localization of both MyoD and Smad3 in CsA-, but not in placebo-, treated mice. Our results demonstrated that CsA treatment upregulates Id1 and Smad3 expression and delays skeletal muscle regeneration in vivo.

Identification of LOV KELCH PROTEIN2 (LKP2)-interacting factors that can recruit LKP2 to nuclear bodies.

LOV KELCH PROTEIN2 (LKP2) is an F-box protein that has been postulated to function centrally, or near to the circadian clock oscillator. As a first step to determine which proteins act as substrates of LKP2, yeast two-hybrid screening was performed using LKP2 as bait, and two interaction factors, Di19 and COL1, were isolated. The transiently expressed Di19-GUS fusion protein was localized in the nucleus of Arabidopsis petiole cells. COL1 and other CO/COL family proteins could also interact with LKP1/ZTL, LKP2 or FKF1. The LKP2-binding site in CO or COL1 was near the center of each protein. The CCT motif in CO or COL1 was not sufficient for interaction with LKP2. LKP2 recognized CO with F-box and kelch repeat-containing regions, while it recognized COL1 with an LOV domain. When LKP2 was fused with cyan fluorescent proein (CFP) and transiently expressed in onion epidermal cells, CFP-LKP2 signals were localized in the nucleus and cytosol. Both yellow fluorescent protein (YFP)-CO and YFP-COL1 were located in the nucleus, forming nuclear bodies when they were transiently expressed. However, co-expression of CFP-LKP2 with YFP fused to either CO or COL1 resulted in the recruitment of CFP-LKP2 in nuclear bodies. Furthermore, the CFP-LKP2 and YFP-CO signals co-localized with signals for pU2B''-mRFP, which is a marker for Cajal bodies. These results suggest the possibility that LKP2 functions with CO/COL family proteins in the nuclear bodies.

Increase of Cardiotrophin-1 immunoreactivity in regenerating and overloaded but not denervated muscles of rats.

The original report by Pennica et al. on Cardiotrophin-1 (CT-1) states that it markedly stimulates hypertrophy in cardiac myocytes both in vitro and in vivo and is predominantly expressed in the early mouse embryonic heart tube. CT-1 is a member of the interleukin-6 superfamily and past studies have shown that it exerts trophic effects on neurons, glial cells and their precursors, and is expressed during myogenesis. Thus CT-1 is associated with physical and pathological changes in skeletal muscle. In this study, we examined whether CT-1 is expressed in mechanically overloaded, regenerating, and denervated muscles of rats using immunohistochemistry. In the overloaded plantaris muscles at 1 and 3 days postsurgery, CT-1 immunoreactivity was detected in the mononuclear cells that had infiltrated the extracellular space. CT-1 immunoreactivity was also observed in the mononuclear cells invading the extracellular space at 2, 4, and 6 days after a bupivacaine injection and in degenerative and necrotic muscle fibers at 2 days postinjection. In the denervated muscles, the CT-1 immunoreactivity did not change in intensity during the entire period of the denervation (2, 7, and 14 days postsurgery). The cells invading extracellular space and in necrotic muscle fibers possessing CT-1 immunoreactivity might be muscle precursor cells (satellite cells) or migrating macrophages undergoing phagocytosis. Using double-immunostainings for anti-CT-1/antic-met, anti-CT-1/ anti-M-cadherin, and anti-CT-1/anti-ED1, we found that satellite cells and macrophages exhibited CT-1 immunoreactivity in the damaged muscles after bupivacaine injection. We therefore believe that CT-1 plays a key role in regeneration and hypertrophy in the skeletal muscle of rats.

Discrepancy between clinical and pathological diagnoses of CBD and PSP.

Progressive supranuclear palsy (PSP) and corticobasal degeneration (CBD) are often clinically confused with each other. Moreover, the discrepancy between clinical and pathological diagnoses of CBD and PSP are still controversial. We report here two atypical cases of PSP and CBD. A 73-year old woman was admitted with right hand rigidity, limb kinetic apraxia and cortical sensory loss. Brain atrophy, hypoperfusion and hypometabolism predominantly in the left frontoparietal lobes indicated CBD clinically. Pathological studies revealed neuronal loss and spongy change without ballooned neurons (BN) in the cerebral cortex. Modified Gallyas-Braak (G-B) staining revealed neurofibrillary tangles (NFTs) and tufted astrocytes, indicating pathological diagnosis of PSP. A 75-year-old man admitted with vertical gaze palsy, neck dystonia, parkinsonism and dementia. Atrophy of the frontal lobes and tegmentum of the midbrain and symmetrical frontal hypoperfusion in SPECT indicated PSP. However, neuronal loss and BN in the frontal lobes and clusters of astrocytic plaques indicated CBD pathologically. The G-B staining was useful for differentiating between CBD and PSP, but our atypical cases bring up a new issue about differential diagnosis of CBD and PSP.

Individual differences in the kinetics of alcohol absorption and elimination : A human study.

The individual differences in alcohol pharmacokinetics were studied using the one-compartment model with first-order absorption and zero-order elimination kinetics in humans. The blood alcohol concentrations (BACs) were simulated by obtained parameters, absorption rate constant (ka), and climination rate constant (ß). The 81 healthy young Japanese volunteers, who had been divided into those without alcohol-induced facial flushing (nonflushers) and those with facial flushing (flushers) according to alcohol patch test results and a questionnaire beforehand, ingested 0.50 g/kg ethanol within 1 minute. Breath alcohol concentrations (BrACs) were measured during absorption and during the elimination period. BACs were obtained based on BrACs. Fifteen percent of subjects exhibited low BAC profile (below 0.4 mg/mL) (first-pass effect [FPE] group), although the majority showed normal BAC profile (normal group). The ka was approximately 5 to 8 (h(-1)) in the normal group without significant difference between nonflushers and flushers, whereas that in the FPE group was significantly smaller than in the normal group. For the normal group, peak BACs were well simulated by the one-compartment model with first-order absorption and zero-order elimination kinetics. A considerable portion of subjects exhibited FPE. Absorption of alcohol from the intestine plays an important role in alcohol pharmacokinetics in humans.

Lewy bodies in the sinoatrial nodal ganglion: clinicopathological studies.

Lewy bodies (LB) are characteristic pathological findings for idiopathic Parkinson disease, and extracranial organs have also been known to exhibit these structures. Clinically, the possible involvement of LB in cardiac dysfunction has attracted attention based on the findings of studies using [123I] metaiodobenzyl guanidine (MIBG) scintigraphy. The purpose of the present study was to investigate the possible involvement of LB in heart disease. A total of 40 autopsy cases consisting of Lewy body disease and Parkinson syndrome were examined. The former were cases with intracranial LB regardless of clinical symptoms, and the latter were cases with parkinsonism but without intracranial LB. The presence of heart disease or an atrial arrhythmia and the results of an MIBG scintigraphy study were clinically examined. The sinoatrial node was examined microscopically and immunohistochemically. The results showed that heart disease and atrial arrhythmia complications were more frequent in cases with Lewy body disease than in cases with Parkinson syndrome and that LB were frequently found in extracranial organs, especially in the sinoatrial nodal ganglion, in cases with Lewy body disease. In the current report, we hypothesized that neuronal changes involving LB in the sinoatrial nodal ganglion may cause arrhythmia and ischemic heart disease as a result of vasoconstriction.

Effects of ethanol on the induction of uncoupling protein-1 (UCP1) mRNA in the mouse brown adipose tissue.

Expression of uncoupling protein-1 (UCP1) is increased by cold acclimation and overfeeding, and reduced in fasting and genetic obesity. It is known that the mitochondrial UCP1 in the brown adipose tissue (BAT) is an important key molecule for non-shivering thermogenesis. On the other hand, ethanol (EtOH) alters thermoregulation in humans and laboratory animals. However, the relationship between EtOH intake and UCP1 expression is not yet clear. Accordingly, the present study employed the technique of real-time quantitative polymerase-chain reaction (PCR) to investigate the effects of EtOH (0.5 or 2.0 g/kg) on the expression of UCP1 mRNA in the mouse BAT. Control mice were injected with the same volume of physiological saline intraperitoneally (IP). IP injection of EtOH (0.5 g/kg) caused a decrease and an increase of the expression of BAT UCP1 mRNA at 1 and 4 hours, respectively. Treatment with EtOH (2.0 g/kg) caused an increases of the expression of BAT UCP1 mRNA at both 2 and 4 hours. BAT UCP1 mRNA levels in both groups increased at 4 hours after EtOH administration. The levels of UCP1 mRNA returned to the control levels by 8 hours after EtOH administration. The expression of BAT UCP1 mRNA was upregulated following EtOH administration, although a lower dose of EtOH initially reduced the expression of UCP1 mRNA in BAT. These findings suggest that EtOH-induced UCP1 mRNA expression in BAT reflects an alteration of the set point of thermogenesis.