Nondestructive Quantitative Analysis of Difficult-to-Measure Radionuclides 107Pd and 99Tc.

Considering the expanding demand for nuclear waste management of the spent nuclear fuel materials in near future, a nondestructive analytical scheme applicable to one of the most difficult-to-measure nuclides 107Pd, which emits no decay γ-rays and whose half-life is too long to be decayed out during a human lifetime, was designed. The scheme consists of a sophisticated instrument capable of the detection of γ-rays by Ge detectors coupled with time-of-flight measurement of neutrons and a high-intensity pulsed neutron beam and can simultaneously perform time-of-flight-coupled prompt γ-ray analysis (TOF-PGA) as well as PGA and neutron resonance capture analysis (NRCA). The analytical capability for simulated samples of the Tc-platinum group metals (Tc-PGMs) obtained by the group-partitioning process of spent nuclear fuels, which contain not only 107Pd but also 99Tc and other difficult-to-measure fission products, was evaluated. It was confirmed that although PGA and NRCA can accurately analyze both nuclides in individual, single substances, only TOF-PGA can analyze 107Pd as well as 99Tc in the Tc-PGM-simulated sample. The TOF-PGA measurement technique can be widely used for the nondestructive analysis of 107Pd and 99Tc in nuclear wastes.

Adenine-related compounds modulate UDP-glucuronosyltransferase (UGT) activity in mouse liver microsomes.

Adenine-related compounds are allosteric inhibitors of UDP-glucuronosyltransferase (UGT) in rat liver microsomes (RLM) and human UGT isoforms treated with detergent or pore-forming peptide, alamethicin.To clarify whether the same is true beyond species, the effects of adenine-related compounds on 4-methylumbelliferone (4-MU) glucuronidation were examined using detergent-treated mouse liver microsomes (MLM).Brij-58 treatment of MLM increased the Vmax and the Michaelis constant, Km, of 4-MU. This study was performed using Brij-58-treated MLM as an enzyme source. ATP- and ADP-inhibited 4-MU glucuronidation. In contrast, AMP caused a 1.5-fold increase in glucuronidation. Oxidised forms, NAD+ and NADP+, potently inhibited 4-MU glucuronidation, whereas the reduced forms, NADH and NADPH, did not. Furthermore, the IC50 values of ATP, ADP, NAD+, and NADP+ were approximately 15 µM.In our previous study, ATP was the strongest inhibitor of UGT activity in RLM. However, in this study, the above-mentioned compounds inhibited 4-MU UGT in a comparable and non-competitive manner. Furthermore, AMP antagonised the inhibitory effects of ATP and ADP.These results suggest that ATP, ADP, NAD+, and NADP+ are common endogenous inhibitors of UGT beyond species.

Fecal metabolite of a gnotobiotic mouse transplanted with gut microbiota from a patient with Alzheimer's disease.

Studies of Alzheimer's disease are based on model mice that have been altered by transgenesis and other techniques to elicit pathogenesis. However, changes in the gut microbiota were recently suggested to diminish cognitive function in patients, as well as in model mice. Accordingly, we have created model mice of the human gut microbiota by transplanting germ-free C57BL/6N mice with fecal samples from a healthy volunteer and from an affected patient. These humanized mice were stably colonized and reproduced the bacterial diversity in donors. Remarkably, performance on Object Location Test and Object Recognition Test was significantly reduced in the latter than in the former at 55 weeks of age, suggesting that gut microbiota transplanted from an affected patient affects mouse behavior. In addition, metabolites related to the nervous system, including γ-aminobutyrate, taurine, and valine, were significantly less abundant in the feces of mice transplanted with microbiota from the affected patient.

Synergistic effect of combining two nondestructive analytical methods for multielemental analysis.

We developed a new analytical technique that combines prompt gamma-ray analysis (PGA) and time-of-flight elemental analysis (TOF) by using an intense pulsed neutron beam at the Japan Proton Accelerator Research Complex. It allows us to obtain the results from both methods at the same time. Moreover, it can be used to quantify elemental concentrations in the sample, to which neither of these methods can be applied independently, if a new analytical spectrum (TOF-PGA) is used. To assess the effectiveness of the developed method, a mixed sample of Ag, Au, Cd, Co, and Ta, and the Gibeon meteorite were analyzed. The analytical capabilities were compared based on the gamma-ray peak selectivity and signal-to-noise ratios. TOF-PGA method showed high merits, although the capability may differ based on the target and coexisting elements.

Sirolimus monotherapy for Kasabach-Merritt phenomenon in a neonate; Case report.

INTRODUCTION AND IMPORTANCE: The Kasabach-Merritt Phenomenon (KMP), characterized by thrombocytopenia and consumptive coagulopathy due to endothelial cell growth in the infantile vascular tumor kaposiform hemangioendothelioma, presents a therapeutic challenge. This case highlights the novel use of sirolimus in a neonate, an approach less explored in this age group. CASE PRESENTATION: A female neonate presented with a right anterior chest mass, progressing to respiratory distress and congestive heart failure. Diagnosed with KMP, she exhibited low platelet count and coagulation abnormalities. Treatment with sirolimus (0.06 mg/day) led to mass reduction, improved bleeding, and a stable tumor after 12 months, without side effects. This case contrasts with existing literature advocating for combination therapy or higher sirolimus concentrations for effective treatment. Yet, our patient achieved favorable outcomes with low-dose monotherapy, suggesting a potentially safer approach in neonates with immature hepatic and renal metabolism. CLINICAL DISCUSSION: This case demonstrates the efficacy of low-dose sirolimus monotherapy in treating KMP in a neonate, challenging current preferences for combination therapies or higher doses. It emphasizes the need for further research into age-specific treatment protocols in KMP, considering the unique metabolic profiles of neonates and infants. CONCLUSION: Sirolimus has demonstrated potential in treating KMP in pediatric patients. While initial results are promising, determining optimal dosages and trough concentrations, especially in neonates and infants, remains essential.

Integrated pathophysiology of schizophrenia, major depression, and bipolar disorder as monoamine axon disorder.

Recent studies provide evidence that similar to early-stage Parkinson's disease, depression is a neurodegenerative disease characterized by the degeneration of monoamine axons. The major difference between the two disorders is that the symptoms of depression become evident without loss of monoamine neurons, while the motor symptoms of Parkinson's disease appear after loss of the cell body. Given that the axonal degeneration of monoamine neurons underlies the pathophysiology of neurological (Parkinson's disease) and neuropsychiatric (depression) diseases, axonal impairment of monoamine neurons is thought to also occur in schizophrenia and bipolar disorder and play a significant role in the pathophysiology of these mental illnesses. The positive symptoms of schizophrenia and manic symptoms of bipolar disorder are known to occur in hyper-monoaminergic states, opposite to depressive symptoms, negative/cognitive symptoms of schizophrenia, and motor disorders of Parkinson's disease, all occurring in hypo-monoaminergic states. Since monoamine axons have the capacity to spontaneously regenerate or sprout in response to damage in the adult brain and sometimes show hyperinnervation due to excessive regeneration/sprouting beyond normal levels, it is possible that schizophrenia and bipolar disorder are disorders that include excessive regeneration/sprouting of monoamine axons leading to hyper-monoaminergic states. Together, based on accumulating data from animal and human studies, the pathophysiology of schizophrenia, major depression, and bipolar disorder is summarized as follows: The degeneration of monoamine axons is associated with the negative and cognitive symptoms of schizophrenia, major and bipolar depression, while hyper-regeneration/sprouting of monoamine axons underlies the positive symptoms of schizophrenia and bipolar mania. The integrated understanding of schizophrenia, major depression, and bipolar disorder as monoamine axon disorder will open the door to the development of new diagnosis and treatment methods for major mental illnesses as well as early-stage Parkinson's disease.

Effect of sample density in prompt γ-ray analysis.

A high-accuracy analytical method is broadly required to obtain reliable research results. Thus, prompt γ-ray analysis (PGA), one of the most accurate non-destructive analytical methods, has been employed in various fields. However, the measurement accuracy of PGA is also known to degrade in hydrogenous samples. The degradation is caused by variation in the measurement sensitivity (counts per milligram) following the change in neutron energy due to scattering with hydrogen nucleus. Number of scatterings is well known to depend on the hydrogen content in a sample. However, considering multiple scatterings, hydrogen density, which has not been taken into account as yet, may also lead to the accuracy degradation. Here, we show the effect of the hydrogen density in PGA by evaluating the measurement sensitivity of samples with the same hydrogen content and different densities. We find that the measurement sensitivity varies by more than 30% depending on the hydrogen density even at the same hydrogen content. The variation is a particularly serious problem for PGA requiring a few percent accuracy in most cases. Additionally, although the variation is apparently observed in hydrogenous samples, the similar phenomenon can occur in other nuclides with a large scattering cross section; it may affect nuclear cross-section measurements using neutrons in such fields as astrophysics and nuclear energy.

Proteomic analysis of proteins expressing in regions of rat brain by a combination of SDS-PAGE with nano-liquid chromatography-quadrupole-time of flight tandem mass spectrometry.

BACKGROUND: Most biological functions controlled by the brain and their related disorders are closely associated with activation in specific regions of the brain. Neuroproteomics has been applied to the analysis of whole brain, and the general pattern of protein expression in all regions has been elucidated. However, the comprehensive proteome of each brain region remains unclear. RESULTS: In this study, we carried out comparative proteomics of six regions of the adult rat brain: thalamus, hippocampus, frontal cortex, parietal cortex, occipital cortex, and amygdala using semi-quantitative analysis by Mascot Score of the identified proteins. In order to identify efficiently the proteins that are present in the brain, the proteins were separated by a combination of SDS-PAGE on a C18 column-equipped nano-liquid chromatograph, and analyzed by quadrupole-time of flight-tandem-mass spectrometry. The proteomic data show 2,909 peptides in the rat brain, with more than 200 identified as region-abundant proteins by semi-quantitative analysis. The regions containing the identified proteins are membrane (20.0%), cytoplasm (19.5%), mitochondrion (17.1%), cytoskeleton (8.2%), nucleus (4.7%), extracellular region (3.3%), and other (18.0%). Of the identified proteins, the expressions of glial fibrillary acidic protein, GABA transporter 3, Septin 5, heat shock protein 90, synaptotagmin, heat shock protein 70, and pyruvate kinase were confirmed by immunoblotting. We examined the distributions in rat brain of GABA transporter 3, glial fibrillary acidic protein, and heat shock protein 70 by immunohistochemistry, and found that the proteins are localized around the regions observed by proteomic analysis and immunoblotting. IPA analysis indicates that pathways closely related to the biological functions of each region may be activated in rat brain. CONCLUSIONS: These observations indicate that proteomics in each region of adult rat brain may provide a novel way to elucidate biological actions associated with the activation of regions of the brain.

Effects of retinoic acids on the dendritic morphology of cultured hippocampal neurons.

Vitamin A-derived retinoic acids (RAs) are known to exert a variety of biological actions, including modulatory effects on cell differentiation and apoptosis. A recent study has demonstrated that 13-cis-RA and all-trans-RA suppressed neurogenesis in the dentate gyrus of the hippocampus in adult mice. The present experiments were performed to see whether 13-cis-RA and all-trans-RA could alter the dendritic morphology of cultured hippocampal neurons via RA receptors: retinoic acid receptor (RAR) and retinoid X receptor (RXR). High doses of 13-cis-RA and all-trans-RA exerted a negative effect on the cultured hippocampal neurons, while a low dose of 13-cis-RA but not all-trans-RA caused a positive effect. The negative changes induced by 13-cis-RA and all-trans-RA were antagonized by RXR antagonists and RAR antagonists, respectively. The positive changes induced by a low dose of 13-cis-RA were blocked by both RXR antagonists and RAR antagonists. These results suggest that RAs at high concentrations cause a negative effect on the dendritic morphology of cultured hippocampal neurons through RA receptors, while RAs at low concentrations exert a positive influence on cultured hippocampal neurons.

13-cis-retinoic acid alters the cellular morphology of slice-cultured serotonergic neurons in the rat.

Retinoids influence cellular processes such as differentiation, proliferation and apoptosis via retinoic acid receptor (RAR) and retinoid X receptor (RXR), and have therapeutic applications in several cancers and dermatologic diseases. Recent reports indicate that depression occasionally occurs in patients using the acne drug Accutane, the active component of which is 13-cis-retinoic acid (13-cis-RA). Although impairment of serotonin (5-HT)-expressing neurons, including morphologic changes, is thought to be associated with depressive symptoms, the effects of 13-cis-RA on 5-HT neurons have not been examined. The present study demonstrated that 13-cis-RA alters the morphology of 5-HT neurons in cultured rat midbrain slices. The 13-cis-RA-induced changes were partially blocked by RXR and RAR antagonists. Furthermore, cotreatment with RAR and RXR agonists altered the morphology of 5-HT neurons to a greater extent than the individual application of each agonist. The morphologic changes were completely blocked by RXR antagonist, whereas RAR antagonist partially blocked the effects. These results suggest that 13-cis-RA exerts its action on slice-cultured 5-HT neurons, at least in part, through specific retinoid receptors. Moreover, RXR has a greater influence on the morphology of 5-HT neurons than RAR. The receptor-mediated actions of 13-cis-RA presented here may provide a clue for further research on depression associated with the use of 13-cis-RA.

Interferon-alpha reduces the density of monoaminergic axons in the rat brain.

Interferon-alpha commonly induces depressive symptoms in clinical populations; however, the mechanism by which this occurs is unclear. Recent studies suggest that the degeneration of axons containing serotonin and noradrenaline is involved in the pathophysiology of depression. The present immunohistochemical study shows that the density of serotonergic axons decreased in the ventral medial prefrontal cortex and amygdala in the interferon-alpha-treated animals. Additionally, interferon-alpha induced decreases in the density of noradrenergic axons in the dorsal medial prefrontal cortex, ventral medial prefrontal cortex, and dentate gyrus. These results support the hypothesis that long-term administration of interferon-alpha causes the degeneration of monoaminergic axons in specific brain regions, which might be associated with depressive symptoms occurring in interferon-alpha-treated patients.

Stress-induced plasticity of monoamine axons.

Central serotonergic (5-HT) and noradrenergic (NA) neurons, which innervate the same regions of the brain, are known to play a crucial role in emotion and mood. These monoamine neurons have a great capacity to alter axonal morphology in response to repeated stress. The morphological responses of 5-HT and NA axons to repeated stress are different, and they sometimes even demonstrate opposite responses (namely, either sprouting or degeneration). Moreover, a morphological interaction also occurs between 5-HT and NA axons during axonal regeneration. This review describes the differential features of axonal plasticity of 5-HT and NA neurons in relation to stress, and discusses the possible roles that the morphological plasticity of 5-HT and NA axons may play in the pathophysiology of depression.

Draft Genome Sequence of Probiotic Lactobacillus acidophilus Strain L-55 Isolated from a Healthy Human Gut.

Probiotic Lactobacillus acidophilus L-55 was isolated from a healthy human gut. Here, we report the draft genome sequence of this organism.

Convergence and interaction of hippocampal and amygdalar projections within the prefrontal cortex in the rat.

The orbital and medial prefrontal cortex (OMPFC) receives inputs from the CA1/subicular (CA1/S) region of the ventral hippocampus and the basolateral nucleus of the amygdala (BLA). Despite many studies about these projections, little is known as to how CA1/S and BLA inputs converge and interact within the OMPFC. Extracellular recordings of single-unit activity in the OMPFC were performed in sodium pentobarbitone-anesthetized rats. OMPFC neurons driven by CA1/S or BLA stimulation were more frequently encountered in the ventral portion of the prelimbic (v-PrL) and infralimbic cortex (IL). OMPFC neurons showing excitatory convergence of both inputs from the CA1/S and BLA were also located predominantly in the v-PrL and IL. The excitatory latencies of these neurons from both the CA1/S and BLA revealed almost identical values. Excitatory responses of OMPFC neurons to CA1/S (or BLA) stimulation were markedly augmented by simultaneous BLA (or CA1/S) stimulation, whereas the inhibitory influence of the BLA (or CA1/S) on CA1/S-induced (or BLA-induced) excitation was apparent when BLA (or CA1/S) stimulation was given 20-40 msec before CA1/S (or BLA) stimulation. Similar results were also observed when reciprocal connections between the CA1/S and BLA were severed to exclude the influences of these connections on one another. From these studies, we concluded that excitatory and inhibitory inputs from the hippocampus and amygdala converge and interact in the v-PrL and IL. Furthermore, the results indicate that simultaneous activation of hippocampal and amygdalar neurons may be important for amplification of OMPFC neuronal activity.

Cortical spreading depression affects Fos expression in the hypothalamic paraventricular nucleus and the cerebral cortex of both hemispheres.

The present experiments were performed to clarify the brain sites whose activity is affected exclusively by cortical spreading depression (CoSD). For this purpose, Fos protein, a product of an immediate early gene, was used as a marker of neuronal activation. Because Fos can be induced by many manipulations such as stress stimuli, we verified CoSD-induced Fos expression by excluding the influence of other factors such as anaesthesia and surgical manipulation. CoSD was induced by applying a KCl solution directly to the dura mater over the cerebral cortex, and Fos expression in the brain was assessed by immunohistochemistry using antibodies against Fos protein. We found that during CoSD, Fos expression was increased specifically in the magnocellular region of the hypothalamic paraventricular nucleus (PVN), as well as in the ipsilateral cortex, whereas reduced Fos expression was observed in both the parvocellular region of the PVN and the whole cortex contralateral to the CoSD site. Consistent with the reduced Fos expression, approximately 40% of neurons in the contralateral cortex revealed a suppression of electrical activity during CoSD. These results suggest that in addition to the ipsilateral cortex, CoSD affects Fos expression exclusively in the PVN and the contralateral cortex.

Effects of repeated stress on regeneration of serotonergic and noradrenergic axons in the cerebral cortex of adult rats.

We examined whether regeneration of serotonergic (5-HT) and noradrenergic (NA) axons might be affected by stress. Neurotoxins to 5-HT or NA axons were injected into the frontal cortex to cause partial denervation. Mild restraint stress (40 min/day) was started 16 days later and given for 14 consecutive days (30-day stressed group). Non-stressed animals were divided into two groups, animals sacrificed at 14 days (14-day control group) and those sacrificed at 30 days (30-day control group) after the toxin injection. In immunohistochemical study, the denervation area of 5-HT axons but not that of NA axons was significantly smaller in the 30-day control group than in the 14-day control group. However, there was no significant difference between the 14-day control and 30-day stressed groups. These findings suggested that regeneration of 5-HT axons occurred earlier than that of NA axons, and that stress exerted inhibitory influence on regeneration of 5-HT axons.

Paradoxical rise in brainstem PO(2) following umbilical cord occlusion in full-term rat fetuses.

Although the fetus experiences severe hypoxia and ischemia during delivery, the fetal brain is protected from hypoxic-ischemic insults by unknown mechanisms. To investigate this phenomenon, fetal asphyxia was induced in pregnant rats by occlusion of the umbilical cord. Rather than producing cerebral hypoxia, the brainstem PO(2) of at-term fetuses increased following umbilical cord occlusion, while brainstem blood flow was markedly reduced. This paradoxical increase in brainstem PO(2) during asphyxia occurred in the majority of at-term fetuses, less frequently in fetuses 1 day prior to term and did not occur in animals following birth. Because occlusion of the umbilical cord prevents maternal delivery of oxygen to the fetus, we propose that the ability to maintain PO(2) is the result of pre-existing fetal stores of oxygen or from de novo generation of oxygen in the fetal brainstem.

Asphyxia induced by umbilical cord occlusion alters glutamatergic and GABAergic synaptic transmission in neurons of the superior colliculus in fetal rats.

Using optical recordings, we studied the effects of asphyxia on intracellular Cl(-) and Ca(2+) concentrations ([Cl(-)]i; [Ca(2+)]i) in the superior colliculus of fetal rats, which were connected via the umbilical cord to the dam. Acute asphyxia was induced by umbilical cord occlusion. The number of fetal superior colliculus neurons showing GABA-mediated increases in [Cl(-)]i (leading to hyperpolarization) following local synaptic electrical stimulation had decreased by 3 h post-asphyxiation, while the number showing GABA-mediated decreases in [Cl(-)]i (leading to depolarization) increased. [Ca(2+)]i rise, which occurred after acute asphyxiation, was antagonized by both non-NMDA and NMDA receptor antagonists. The increase in [Ca(2+)]i following focal superior colliculus stimulation was markedly attenuated at 3 h post-asphyxiation. These findings suggest that asphyxia induced by umbilical occlusion induces changes in glutamatergic and GABAergic synaptic transmission in the fetal brain.

Exposure to 1-bromopropane causes degeneration of noradrenergic axons in the rat brain.

1-Bromopropane (1-BP) has been used as an alternative to ozone-depleting solvents. Previous studies showed that 1-BP is neurotoxic in animals and humans. In humans, exposure to 1-BP caused various neurological and neurobehavioral symptoms or signs including depressive or irritated mood. However, the neurobiological changes underlying the depressive symptoms induced by 1-BP remain to be determined. The depressive symptoms are thought to be associated with degeneration of axons containing noradrenaline and serotonin. Based on this hypothesis, the present study examined the effects of repeated exposure to 1-BP on serotonergic and noradrenergic axons. Exposure to 1-BP induced dose-dependent decreases in the density of noradrenergic axons in the rat prefrontal cortex, but no apparent change in the density of serotonergic axons. The results suggest that depressive symptoms in workers exposed to 1-BP are due, at least in part, to the degeneration of noradrenergic axons in the brain.

Gene expression profiling in progressively MPTP-lesioned macaques reveals molecular pathways associated with sporadic Parkinson's disease.

Parkinson's disease (PD) is a common neurodegenerative disease characterized by progressive loss of midbrain dopaminergic neurons. To gain an insight into the mechanisms underlying the progression of PD, gene expression analysis was performed using two different brain regions, the substantia nigra pars compacta (SN) and the striatum (STR), of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-lesioned monkey model of PD. 230 genes were differentially expressed in the MPTP-treated SN compared to control, whereas 452 genes showed altered expression in the MPTP-treated STR, implying that MPTP elicits more damages in the striatal gene expression than in the SN. Comparative data analysis of the transcription profiles on the PD patients and MPTP monkey models, and pathway analysis indicated several signaling pathways as possible routes to MPTP-induced neurodegeneration. Interestingly, the networks which associated with cytoskeletal stability, ubiquitin-proteasome system (UPS) and Wnt signaling gained prominence in our study. Further transcriptional regulatory network analysis suggested the association of the neuronal repressor REST (RE1-silencing transcription factor; NRSF) and androgen receptor with the dysregulation of the striatal genes. Our study suggests the possibility that the dysfunction of multi-network signaling may induce abnormalities in a diverse range of biological processes, such as synaptic function, cytoskeletal stability, survival and differentiation.