Improving the depth resolution of STEM-ADF sectioning by 3D deconvolution.

Although the possibility of locating single atom in three dimensions using the scanning transmission electron microscope (STEM) has been discussed with the advent of aberration correction technology, it is still a big challenge. In this report we have developed deconvolution routines based on maximum entropy method (MEM) and Richardson-Lucy algorithm (RLA), which are applicable to the STEM-annular dark-field (ADF) though-focus images to improve the depth resolution. The new three-dimensional (3D) deconvolution routines require a limited defocus-range of STEM-ADF images that covers a whole sample and some vacuum regions. Since the STEM-ADF probe is infinitely elongated along the optical axis, a 3D convolution is performed with a two-dimensional (2D) convolution over xy-plane using the 2D fast Fourier transform in reciprocal space, and a one-dimensional convolution along the z-direction in real space. Using our new deconvolution routines, we have processed simulated focal series of STEM-ADF images for single Ce dopants embedded in wurtzite-type AlN. Applying the MEM, the Ce peaks are clearly localized along the depth, and the peak width is reduced down to almost one half. We also applied the new deconvolution routines to experimental focal series of STEM-ADF images of a monolayer graphene. The RLA gives smooth and high-P/B ratio scattering distribution, and the graphene layer can be easily detected. Using our deconvolution algorithms, we can determine the depth locations of the heavy dopants and the graphene layer within the precision of 0.1 and 0.2 nm, respectively. Thus, the deconvolution must be extremely useful for the optical sectioning with 3D STEM-ADF images.

Fos expression in neurons immunoreactive for neuronal nitric oxide synthase in the rat paraventricular nucleus after intraperitoneal injection of interleukin-1 beta.

Double immunostaining for Fos and neuronal nitric oxide synthase (nNOS) was used to examine whether nNOS-immunoreactive neurons in the paraventricular hypothalamic nucleus (PVN) are activated to express Fos immunoreactivity by intraperitoneal injection of interleukin-1 beta (IL-1 beta) in the rat. Quantitative analysis revealed that some nNOS-positive PVN neurons are activated by IL-1 beta (4 microg/kg, i.p.) administration, but the majority of the IL-1 beta-activated PVN neurons do not express nNOS and are distributed mainly in the parvocellular part of the PVN.

Effect of intra-amygdala dopaminergic grafts on methamphetamine-induced locomotor activity, extracellular dopamine and dopamine metabolite overflow: a comparison with the effect of intra-accumbens grafts.

The effects of 6-hydroxydopamine lesions of the ventral tegmental area and following intra-amygdala or intra-accumbens dopaminergic (DAergic) grafts on methamphetamine (MAP)-induced locomotor activity were investigated in rats. Intra-accumbens DAergic grafts from rat embryos restored the locomotor hyperactivity response to MAP 5 weeks after grafting, while intra-amygdala grafts did not restore responses by 10 weeks after grafting. Biochemical measurements of extracellular DAergic activity in the amygdala (AMY) by in vivo microdialysis after grafting showed no significant change in the basal levels of dopamine (DA) and partial restoration of metabolite levels. MAP induced an increase of DA efflux and a decrease in dihydroxyphenylacetic acid without a significant change in homovanillic acid, which is the same pattern of response seen in control animals. These biochemical changes are similar to those seen previously after intra-accumbens grafts. The results show that restoration of DAergic activity in the AMY in the presence of DAergic denervation of accumbens does not have an effect on MAP-induced locomotion.

Dopaminergic transplants suppress L-DOPA-induced Fos expression in the dopamine-depleted striatum in a rat model of Parkinson's disease.

We examined the effects of MK-801, a non-competitive antagonist of the N-methyl-D-aspartate (NMDA) receptor, and fetal ventral mesencephalic (VM) transplants on L-3,4-dihydroxyphenylalanine (L-DOPA)-induced Fos protein expression in the dopamine (DA)-depleted striatum. Unilateral 6-hydroxydopamine (6-OHDA) lesions of the nigrostriatal pathway were produced in young adult female rats and grafting was performed 3 weeks later. Methamphetamine-induced rotational behavior recovered significantly on the 4th week after grafting. Immunohistochemical examinations of c-Fos and tyrosine hydroxylase (TH) were performed 3-4 months after grafting. L-DOPA (100 mg/kg, i.p.) markedly induced Fos-like immunoreactivity (FLI) in the DA-depleted striatum. Pretreatment with a large dose of MK-801 (3-4.5 mg/kg, i.p.) dose-dependently suppressed L-DOPA-induced FLI in the striatum. The stimulatory effect of L-DOPA on c-Fos expression observed within the lesioned striatum was suppressed by fetal VM transplants. It seemed that the graft-induced effect on FLI extended over a considerably larger area than that covered by the graft-derived TH-immunoreactive innervation. Taken together, these findings suggest that glutamatergic modulation is involved in the L-DOPA-induced c-Fos expression in the denervated striatum which is normalized by fetal VM transplants. It also seems likely that VM grafts suppress the L-DOPA-induced expression of transcriptional factors which might be involved in the mechanisms underlying various side effects of chronic L-DOPA therapy.

Methamphetamine induces fos expression in the striatum and the substantia nigra pars reticulata in a rat model of Parkinson's disease.

In rats with a unilateral 6-hydroxydopamine (6-OHDA) lesion in the nigrostriatal pathway, methamphetamine (3 mg/kg, i.p.) induced Fos-like immunoreactivity (FLI) not only in the striatum on the intact side but also in the substantia nigra pars reticulata (SNr) on the lesioned side. The methamphetamine-induced hyperexpression of FLI in the SNr on the lesioned side was suppressed by pretreatment with either dopamine D1 receptor antagonist SCH-23390 (0.5 mg/kg, i.p.), D2 receptor antagonist raclopride (2 mg/kg, i.p.) or N-methyl-d-aspartate receptor antagonist MK-801 (1 mg/kg, i.p.), which was concomitant with inhibition of the methamphetamine-induced rotational behavior of each antagonist. However, the hyperexpression of FLI in the SNr was not suppressed by intrastriatal grafts of fetal ventral mesencephalon which could suppress the methamphetamine-induced rotation completely. These results indicate that opposite hemispheric asymmetries in FLI are induced by methamphetamine in the striatum and the SNr in the 6-OHDA rats. It is suggested that the FLIs in the two discrete sites are activated independently by different mechanisms, and furthermore, different neuronal pathways are involved in the methamphetamine-induced rotation and Fos expression in the SNr of 6-OHDA rats.

Serotonergic activity in the rat striatum after intrastriatal transplantation of fetal nigra as measured by microdialysis.

In vivo microdialysis was used to examine the effects of dopaminergic transplants on extracellular concentrations of dopamine (DA), serotonin (5-HT), and their precursors and major metabolites in the denervated rat striatum. Dialysis perfusates were collected from intact 6-hydroxydopamine (6-OHDA) lesion plus sham grafted, and lesion plus fetal substantia nigra (SN) grafted striata. The SN transplants ameliorated the reduction of striatal DA and dihydroxyphenylacetic acid (DOPAC) levels in rats with unilateral 6-OHDA lesions of the mesostriatal pathway. The transplants also increased extracellular levels of 5-HT and 5-hydroxyindoleacetic acid (5-HIAA) in the denervated striatum. In response to NSD-1015 (an inhibitor of aromatic L-amino acid decarboxylase, AADC), 5-hydroxytryptophan (5-HTP) levels were substantially elevated in the SN grafted striata as compared with those in the sham grafted controls, which continued even after subsequent administration of L-3,4-dihydroxyphenylalanine (L-DOPA, 100 mg/kg i.p.). Immunohistochemical analysis showed hyperinnervation of 5-HT fibers in the grafted striatum, which was consistent with the results of microdialysis experiments. These results indicated that implantation of SN grafts into the 6-OHDA-lesioned striatum of rats induces hyperactivity of 5-HT synthesis, release and metabolism.

Repeated administration of high dose levodopa enhances hydroxyl radical production in the rat striatum denervated with 6-hydroxydopamine.

We examined whether levodopa (L-DOPA) might increase production of hydroxyl radicals in intact and dopamine-denervated rat striatum. Salicylate trapping combined with in vivo microdialysis provided measurements of 2,3-dihydroxybenzoic acid (2,3-DHBA) as a marker of hydroxyl radical production. Acute administration of high-dose L-DOPA (200, 500 mg/kg, i.p.) did not alter 2,3-DHBA levels in intact striatum or in striatum denervated with 6-hydroxydopamine. On the other hand, L-DOPA administration (200 mg/kg, i.p.) transiently increased 2,3-DHBA in dopamine-denervated striatum of rats after repeated administration of L-DOPA (200 mg/kg, i.p., once daily for 16 days). The results indicated that repeated administration of high dose L-DOPA increased production of hydroxyl radicals in dopamine-denervated striatum.

Dopaminergic transplants alter in vivo activity of tryptophan hydroxylase in the striatum in a rat model of Parkinson's disease.

L-3,4-Dihydroxyphenylalanine (L-DOPA) inhibits the activity of tryptophan hydroxylase (TRH) and thus serotonin synthesis. This inhibitory effect of L-DOPA may be related to some side effects in the patients under L-DOPA therapy. The effects of transplantation of fetal ventral mesencephalon (VM) on extracellular 5-hydroxytryptophan (5-HTP) accumulation was examined by microdialysis as an index of in vivo activity of TRH in the striatum of 6-hydroxydopamine (6-OHDA)-lesioned rats. In the rat striatum perfused with m-hydroxybenzylhydrazine (NSD-1015; an inhibitor of aromatic L-amino acid decarboxylase), L-DOPA and 5-HTP in dialysate were measured simultaneously. In response to NSD-1015, 5-HTP levels were substantially elevated in the lesion plus VM-grafted striata as compared with those in the lesion plus sham-grafted striata. The results indicate that implantation of dopamine-rich VM grafts into the 6-OHDA-lesioned striatum of rats induces hyperactivity of TRH.

Peripherally administered tetrahydrobiopterin increases in vivo tryptophan hydroxylase activity in the striatum after transplantation of fetal ventral mesencephalon in six hydroxydopamine lesioned rats.

The intraperitoneal administration of 6R-L-erythro-5,6,7,8-tetrahydrobiopterin (6R-BH4), a natural cofactor for tyrosine hydroxylase and tryptophan hydroxylase (TRH), dose-dependently increased the extracellular concentration of 6R-BH4 itself in rat striatum. The concentration was investigated by in vivo microdialysis and measured simultaneously with 5-hydroxytryptophan (5-HTP), a precursor of serotonin, by high performance liquid chromatography with electrochemical detection. The 6R-BH4 (50 mg/kg, i.p.) administration increased the accumulation of 5-HTP as an index of in vivo TRH activity under the inhibition of aromatic L-amino acid decarboxylase by NSD-1015 in the striatum of both normal control and 6-hydroxydopamine lesioned rats with intrastriatal transplants of fetal ventral mesencephalon (VM). The results suggest that TRH in the striatum of both control and VM-grafted rats is activated by 6R-BH4 penetrating into the brain from the blood.

Selective release of serotonin by endogenous alkaloids, 1-methyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinolines, (R)- and (S)salsolinol, in the rat striatum; in vivo microdialysis study.

Dopamine-derived 1-methyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinolines, (R)- and (S)salsolinol, released an enormous amount of serotonin in the rat striatum; the concentration of serotonin increased from undetectable level to 2.53 +/- 0.12 and 3.69 +/- 0.01 microM after perfusion of (R)- and (S)salsolinol, respectively. Salsolinols increased extracellular dopamine level, but to a much lesser degree than serotonin. Other naturally occurring isoquinolines with catechol structure released serotonin and dopamine, but salsolinols were the most potent and selective releaser of serotonin. Serotonin release by salsolinols may be involved in some psychiatric symptoms in L-DOPA therapy or alcoholism.

A Japanese case of Creutzfeldt-Jakob disease with a point mutation in the prion protein gene at codon 210.

We screened 111 cases of sporadic Creutzfeldt-Jakob disease (CJD) and 75 healthy control subjects in Japan to detect possible polymorphisms in their prion protein gene (PRNP). We identified a G-to-A point substitution at codon 210, leading a valine-to-isoleucine change, in a 69-year-old CJD patient. This substitution was not seen in 75 healthy control subjects.

Heightened IgE response to mite antigens in inflammatory neuropathies.

In order to clarify the IgE response to common environmental antigens, we measured the serum total IgE and allergen-specific IgE in 50 patients with Guillain-Barré syndrome (GBS), nine patients with Fisher syndrome (FS), 14 patients with chronic inflammatory demyelinating polyneuropathy (CIDP), 18 patients with mononeuritis multiplex (MNM), 43 patients with neurodegenerative disorders and 82 healthy controls by ELISA. The total IgE level was significantly higher in patients with GBS (median = 135 U/ml, P<0.05), CIDP (median = 175 U/ml, P<0.05) and MNM (median= 199 U/ml, P<0.05), than in the healthy controls (median = 79 U/ml), but not in those patients with neurodegenerative disorders. The specific IgE to Dermatophagoides pteronyssinus was significantly higher in the patients with GBS (56%, P<0.01) and MNM (72%, P<0.005) than in the healthy controls (32%). The level of specific IgE to Dermatophagoides farinae tended to be higher in the patients with GBS than in the healthy controls (0.05

Single and repeated environmental stress: effect on plasma oxytocin, corticosterone, catecholamines, and behavior.

Rat studies were done to further characterize an environmental model of stress designated shaker stress (SS). Plasma oxytocin (OT), corticosterone (CS), norepinephrine (NE), and epinephrine (E) were measured before and after 5 or 30 min of SS applied one time or applied 10 times over a 2-week period. The major findings were partial adaptation of plasma E within 30 min of acute SS, adaptation of plasma CS baselines but not responses to chronic SS, and complete adaptation of plasma OT responses to chronic SS. Poststress behavior during chronic SS was affected in the following ways: freezing time habituated, defecation and rearings increased, and grooming and teeth chattering remained relatively constant. The results show that SS produces consistent patterns of hormonal and behavioral responses; some aspects of the patterns are similar to those elicited by other environmental stresses, whereas some aspects are unique to SS. We conclude that rats do not adapt to repeated SS but rather that most hormonal and behavioral defense mechanisms are renewable on a daily basis.

Differentiation of Herpes simplex virus types 1 and 2 in sera of patients with HSV central nervous system infections by type-specific enzyme-linked immunosorbent assay.

OBJECTIVE: To differentiate by enzyme-linked immunosorbent assay (ELISA) using type-specific glycoprotein G herpes simplex virus (HSV) type 1 and type 2 in serum collected from patients with HSV central nervous system (CNS)infections. METHODS: HSV 1 and 2 typing in convalescent sera of 17 patients with HSV acute encephalitis, myelitis, or meningitis was determined by the type-specific IgG ELISA kit (Gull Laboratory, Inc.). HSV CNS infections were diagnosed by polymerase chain reaction (PCR) or conventional serologic tests from acute to convalescent stages. RESULTS: In 13 of 17 patients, HSV type 1 and HSV type 2 antibodies were positive; 11 patients with HSV type 1 and 2 patients with HSV type 2 were found. All 10 patients with encephalitis showed equivocal or positive results for HSVtype 1. In two of three cases of myelitis, HSV type 1 was demonstrated. Each case of myelitis and meningitis reacted to both types 1 and 2. CONCLUSION: These data suggest that the kit is useful for type differentiation of HSV CNS infections from convalescent sera, and can play a supplementary role in HSV typing by PCR or previous serologic tests.

Septic arthritis of the hip associated with atopic dermatitis. A case report.

We report a case of septic arthritis of the hip associated with atopic dermatitis. A 15-year female felt a pain in the right hip with unknown cause on May 11, 1998. The pain subsequently became aggravated, and she was admitted to our hospital on May 18. She has had atopic dermatitis since 4 years of age. She showed generalized dermatitis with desquamation and numerous scratch marks. A culture of both skin and joint fluid revealed Staphylococcus aureus. Physical examination revealed tenderness in Scarpa triangle and restricted range of motion. Immunological serology showed an increase in eosinophils and immunoglobulin E, and a decreased reaction of lymphocyte blastoid transformation. Computed tomography (CT) and MRI showed a joint effusion in the right hip. She was diagnosed as having septic arthritis of the hip. Intravenous drip of Cefazolin of 2g was started on the first day of hospitalization and joint irrigation was done on the second day. CRP became negative at 4 weeks, but joint effusion was shown on CT. Additional joint irrigation with Amicamycin (200 mg) was done. As the joint fluid culture became negative, range of motion exercises were started at 6 weeks. She was discharged with a long-leg brace applied at 8 weeks. At 13 months after onset, she had complete relief of the pain and normal activities of daily living. No destructive changes in the hip were found on X-ray examination or MRI. In the present case, an abnormal immune system associated with atopic dermatitis as well as the habit of scratching eruptions may have led to hematogenous spread of skin infection, and caused septic arthritis of the hip.

Transient osteoporosis of the hip during pregnancy.

We report the clinical features of and MRI findings in transient osteoporosis of the hip during pregnancy. The study population consisted of 4 patients with a mean age of 33 years. The mean gestational age at onset was 31 weeks (range: 27 to 35 weeks). The main symptoms consisted of a weight-bearing pain in the hip and gait disturbance. The pain occurred suddenly and was of unknown cause and became severe within 2 to 3 weeks. X-ray examinations showed diffuse osteoporosis in the femoral head and neck. Moreover in 3 patients, similar lesions were also found in the lumbar spine or the knee. MRI obtained from 3 patients revealed a mottled low-signal lesion extending from the femoral head and neck on T1-weighted images and a high-signal lesion in the bone marrow suggesting edema on T2-weighted images. Mild elevation of C- reactive protein was shown in 2 patients. Conservative treatments with the limitation of weight bearing and bed rest were performed for all patients, and nonsteroidal anti-inflammatory drugs were given to 3 patients. The hip pain began to decline from 8 to 14 weeks after the onset, and completely disappeared from 14 to 24 weeks. X-ray examinations showed that osteoporotic lesions tended to improve at 10 to 14 weeks, on MRI, a high-signal lesion suggesting bone marrow edema resolved together with relief of the pain. No recurrence was found in any patients at mean follow-up of 70.8 months.

A suspension procedure using the extensor carpi ulnaris tendon for distal radioulnar joint disorders.

Recently the Sauve-Kapandji (S-K) procedure has become popular for the treatment of various distal radioulnar joint (DRUJ) disorders. However, some complications, especially pain over the proximal stump of the ulna due to instability of the ulna have been reported in more recent follow-up studies. To prevent the occurrence of this pain, we devised a modified S-K procedure, which we called the suspension procedure, in which the extensor carpi ulnaris (ECU) tendon was used to suspend the proximal ulnar stump. We report here the surgical technique and compare clinical and radiographic results between the suspension procedure and the S-K procedure alone. We performed the S-K procedure alone on 8 patients (original group) and the suspension procedure on 5 (suspension group). Clinical results were assessed according to the clinical evaluation scoring system described by Inoue. Radiographic evaluations included the radio-ulnar distance, the gap of the ulna, and the distance between the articular surface of the wrist and the proximal ulnar stump. In the original group, 4 patients were rated as excellent, 2 as good and 2 as fair, whereas in the suspension group, 3 were rated as excellent, 2 as good and none as fair. In regard to radiographic evaluations, there were no significant differences in any of the 3 parameters between the 2 group. This suspension procedure had an advantage over the S-K procedure alone, especially in preventing the occurrence of stump pain. As there was no significant difference in radiographic findings between the two procedures regarding the site of osteotomy, the amount of bone resection, and radio-ulnar distance, stump pain may be attributed to dynamic instability rather than to static instability.

[A case of portal-systemic encephalopathy associated with Sheehan's syndrome].

A 66-year-old woman had had recurrent episodes of disturbed consciousness whenever she had been constipated or dehydrated. She had been inactive and afflicted with obstinate constipation since she had menopause at age of 32. She underwent gastrectomy for gastric ulcer at age of 37. Laboratory examination showed marked hyperammonemia, reduction in Fisher ratio, and poor excretion of ICG. Furthermore, hypopituitarism and secondary hypothyroidism were found. She was diagnosed as Sheehan's syndrome. A T1-weighted MRI demonstrated symmetrical high intensity in the bilateral globus pallidus and empty sella. The histological examination of the liver revealed a mild lymphocytic infiltration without liver cirrhosis. Abdominal angiography showed a large shunt vessel between the splenic vein and the left renal vein. After embolization of the shunt vessel, hyperammonemia and neurological impairment improved. Additionally multiple hormones replacement has been useful to reduce the drugs of standard therapy for hepatic coma. In this case, we speculated that Sheehan's syndrome accelerated the constipation and worsened the shunt encephalopathy.

[Fibronectin (FN) concentrations in plasma and cerebrospinal fluid (CSF) from guinea pigs with experimental allergic encephalomyelitis (EAE) induced by myelin basic proteins].

The aim of this study was to ascertain whether the FN concentrations in plasma and in CSF are related to the symptomatic status of EAE induced in guinea pigs by myelin basic protein. Guinea pigs were immunized with myelin basic proteins in Freund's complete adjuvant, and after the appearance of neurological symptoms, the plasmas and CSFs from these animals were individually collected. The FN concentrations in these specimens were determined by a solid-phase inhibitory radioimmunoassay using a rabbit antibody specific for guinea pig FN. In plasmas from EAE induced animals, the average value of FN concentrations was lower than that from control animals, but in CSFs from EAE induced animals the average value was slightly higher than that from control animals. The FN concentration in plasma from individual animals with or without EAE was not related to that in the respective CSF, and no direct correlation between the symptomatic severity of EAE and the FN concentration in CSF from the respective animals was observed. These results indicate that the FN concentrations in CSFs probably increase in association with the induction of EAE in guinea pigs, but the levels are highly variable in individual cases, and that the FN concentration in CSF is not available to use as a consistent indicator for EAE in guinea pigs.

Nuclear pore changes and absence of apoptosis in lumbar dorsal root ganglion neurons of doxorubicin-intoxicated rats.

Doxorubicin (DXR) produces degeneration of neurons in the lumbar dorsal root ganglion (DRG) in rats. Light microscopic studies, which included the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling method, and electron microscopic observation revealed that the moderate nuclear and remarkable cytoplasmic degeneration of DRG neurons of Sprague-Dawley rats after intravenous administration of 8 mg/kg of DXR was cell necrosis, not apoptosis. In some neurons, mostly dark and usually with moderate degrees of nuclear degenerative changes, the nuclear pores were decreased in number and obscure 14 and 20 days after DXR administration. DXR enters presumably the nucleus and is partly removed through the nuclear pores. However, the diameters of nuclear pores were similar in DXR-intoxicated and control rats. The changes in nuclear pores of neurons in DXR intoxication, which to our knowledge has not been previously studied, are considered to be part of the degenerative or necrotic changes of DRG neurons.