Molecular monitoring and isolation of previously uncultured bacterial strains from the sheep rumen.

To estimate the contribution of uncultured bacterial groups to fiber degradation, we attempted to retrieve both ecological and functional information on uncultured groups in the rumen. Among previously reported uncultured bacteria, fiber-associated groups U2 and U3, belonging to the low-GC Gram-positive bacterial group, were targeted. PCR primers and fluorescence in situ hybridization (FISH) probe targeting 16S rRNA genes or rRNA were designed and used to monitor the distribution of targets. The population size of group U2 in the rumen was as high as 1.87%, while that of group U3 was only 0.03%. Strong fluorescence signals were observed from group U2 cells attached to plant fibers in the rumen. These findings indicate the ecological significance of group U2 in the rumen. We succeeded in enriching group U2 using rumen-incubated rice straw as the inoculum followed by incubation in an appropriate medium with an agent inhibitory for Gram-negative bacteria. Consequently, we successfully isolated two strains, designated B76 and R-25, belonging to group U2. Both strains were Gram-positive short rods or cocci that were 0.5 to 0.8 mum in size. Strain B76 possessed xylanase and alpha-l-arabinofuranosidase activity. In particular, the xylanase activity of strain B76 was higher than that of xylanolytic Butyrivibrio fibrisolvens H17c grown on cellobiose. Strain R-25 showed an alpha-l-arabinofuranosidase activity higher than that of strain B76. These results suggest that strains B76 and R-25 contribute to hemicellulose degradation in the rumen.

Study of edema reduction patterns during the treatment phase of complex decongestive physiotherapy for extremity lymphedema.

Shortening the treatment phase of complex decongestive physiotherapy (CDP) is extremely important both for individual patients and medical economics. In 83 patients with stage II unilateral secondary extremity lymphedema (31 upper extremities and 52 lower extremities), the daily changes in the volume of affected extremities during the treatment phase of CDP were prospectively investigated. For the upper extremity lymphedemas, the biggest change was seen between days 1 (100% residual edema rate) and 2 (46.0 +/- 2.7%; mean +/- SD) of therapy with a 54.0% reduction (p < 0.0001). Between days 2 and 3 (38.0 +/- 2.6%) of therapy, there was an 8.0% reduction (p < 0.05). From days 3 to 6 of therapy, slight changes ranging from 0.2 to 3.2%/day were seen. For the lower extremity lymphedemas, the biggest change was seen between days 1 (100%) and 2 (44.5 +/- 2.1%) of therapy with a 55.5% reduction (p < 0.0001). Between days 2 and 3 (33.5 +/- 2.6%) of therapy, there was an 11.0% reduction (p < 0.001). The daily volume changes from days 4 to 6 were slight, ranging from 0.1 to 1.0%/day. During the treatment phase of CDP, the largest volume changes were seen soon after the start of therapy.

High-grade broad ligament cancer of müllerian origin: immunohistochemical analysis of a case and review of the literature.

Malignant neoplasms arising in the broad ligament are quite unusual. Although tens of broad ligament cancers of low potential malignancy have been documented, there have been only two reports of high-grade cases. Furthermore, it remains controversial whether broad ligament tumors are mesonephric (wolffian) or paramesonephric (müllerian) in origin. This case study reports a 49-year-old nulliparous woman who, during her follow-up for breast cancer, demonstrated a 39- x 32-mm solid and poorly differentiated adenocarcinoma within the broad ligament of the right adnexa. Because of its rarity and poor differentiation, immunohistochemical analysis was performed to identify the primary malignancy and determine its origin. Positive expression of WT1, CA125, and ER and negative expression of GCDFP-15, HER-2, and PgR in broad ligament cancer, in conjunction with positive expression of GCDFP-15 and ER and negative expression of WT1, HER-2 and PgR in the previous breast cancer, suggested broad ligament cancer to be a gynecological primary malignancy with serous characteristics rather than a metastasis from the previous breast cancer. In addition, EMA-positive and CK7-negative expression indicated a müllerian origin. Broad ligament cancers of müllerian origin occur predominantly in women of reproductive age, most of whom are infertile or nulliparous. This feature, together with the positive expression of ER in this case, may raise the possible hormonal influence on the etiology of broad ligament cancer of müllerian origin.

Endovascular therapy followed by stereotactic radiosurgery for cerebral arteriovenous malformations.

SUMMARY: Pre-radiosurgical embolization was carried out using cyanoacrylate in seven of 13 patients with cerebral arteriovenous malformations (AVMs) treated by stereotactic radiosurgery (SRS) with a linear accelerator (LINAC). The aim of embolization before SRS was the reduction of AVM volume and/or the elimination of vascular structures bearing an increased risk of haemorrhage. Staged-volume SRS was also performed in two patients because of residual irregular shaped nidus of AVMs even after the embolizations. Complete obliteration of the AVM nidus on angiogram was presented in five patients with embolizations (including one with staged-volume SRS) and in three of six patients with SRS alone, during follow-up periods after radiosurgery. No patients experienced haemorrhagic events after SRS. Although transient neurological symptoms were observed after embolizations in two patients, no permanent neurological deficits were presented in all patients with SRS. Pre-radiosurgical embolization may allow the effective influence on irradiation therapy in relatively large AVMs and promote more frequent obliteration in more small sized AVMs compared to those with SRS alone. However, further study must be needed to determine whether staged-volume SRS provides a high rate of AVM obliteration and its safeness.

Single burr hole surgery for the spheno-orbital fibrous dysplasia using intraoperative computed tomography.

Total removal of spheno-orbital fibrous dysplasia was achieved through intraoperative CT-assisted surgery via a burr hole. A 32-year-old man had persistent headache. Radiological studies demonstrated a small osteolytic lesion in the sphenoidal bone underneath the superior orbital fissure. Intraoperative serial CT scans showed the depth and width of the tumor within the complicated structure of the skull base. The lesion was successfully removed by CT-guided minimally invasive surgery.

Dural Arteriovenous Fistula within the Hypoglossal Canal Successfully Treated by Transvenous Embolization. Case Report.

SUMMARY: We report a case of dural arteriovenous fistula (DAVF) within the left hypoglossal canal in a 64-year-old man who presented with tinnitus and ocular symptoms. Angiography revealed DAVF with the fistulous pouch medial to the left jugular bulb. The fistula was feeded by meningeal branches of the bilateral ascending pharyngeal arteries and the branches from the left vertebral artery. The fistula shunted into the left jugular bulb, with reflux into the left inferior petrosal (IPS) and cavernous sinuses (Cses), left superior ophthalmic vein (SOV) and cortical veins over the cerebral convexity. We performed transvenous coil embolization to occlude the fistula resulting in complete resolution of symptoms and signs.

Frameless stereotactic radiosurgery with mobile CT, mask immobilization and micro-multileaf collimators.

We present frameless stereotactic radiosurgery using mobile CT, thermal plastic mask fixation, a vacuum-form body immobilizer and micro-multileaf collimators. A linear accelerator and a self-moving helical CT scanner gantry were installed in the same room. The isocenter of irradiation can be also aligned with the center of the CT gantry by rotating the couch. A thermal plastic mask and vacuum-form body immobilizer was used for registration and immobilization. The advantages of this system are as follows: 1) Accurate and painless frameless fractionated irradiation can be performed smoothly, as the patient's head is fixed without exchanging the couch from CT scanning to irradiation system. 2) This mask system can be applied to children, infants or adults with a previous craniotomy bone flap that must have a fixation pin placed into it. This system can be also used for fractionated radiotherapy without painful skull pin-fixed frame. 3) 1 mm micro-multileaf collimators enable irregular contour irradiation. 4) Image fusion (among CT, MRI, angiography, and PET) and 3D images can be used for irradiation planning. 5) This system can be used on any part of the body. 6) This system can be installed in any irradiation room without any extension or new construction.

[Contrivance for frozen section diagnosis of early minute lung cancer during operation].

We reported the technique of pathological diagnosis for minute lung cancer lesion of early stage during operation. Target lesion for our technique may be less than 1 cm in size and not be detected by palpation. Our technique may superior to usual method of frozen section of tissue specimen without fixation. At first, resected lung specimen was fixed by injection of 20% formalin solution subsequent warming by microwave oven for 40 seconds. Almost all lesions may be detected in proper thickness of specimen. Specimen was washed out and steeped in the 20% of sucrose solution for 30 to 60 seconds in order to avoid crystallization of water that was contained within lung specimen. Then thin sliced section of specimen was made by Cryostat in usual method. The minute cancer lesion was able to detect more easy than usual method of making frozen section of specimen. Formalin fixed and sucrose steeped frozen section may be able to evaluate the surgical margin from cancer exactly than the section made by non-fixed specimen.

Ultrastructure of NADPH diaphorase-positive nerve fibers and their terminals in the rat cerebral arterial system.

To investigate how perivascular NO synthase (NOS)-containing nerves in the cerebral arterial system are involved in controlling the cerebral circulation, we observed the ultrastructure of NOS-containing nerve fibers and their terminals by means of nicotinamide adenine dinucleotide hydrogen phosphate-diaphorase (NADPH-d) histochemistry. We also observed the correlation between NADPH-d stained perivascular nerves and the perivascular sympathetic nerves, by means of double staining with NADPH-d histochemistry and tyrosine hydroxylase (TH) immunohistochemistry at the light microscopic level. NADPH-d-positive nerve fibers showed dense distribution mainly in the rostral portion of the circle of Willis and proximal portions of its main branches, where some of the NADPH-d-positive fibers coexisted with TH-positive fibers in a single nerve bundle. NADPH-d-positive nerve fibers were unmyelinated and had close contact with NADPH-d-negative myelinated and unmyelinated nerve fibers in a single nerve bundle, and NADPH-d-positive nerve terminals also existed closely with NADPH-d-negative nerve terminals. The number of NADPH-d-positive nerve terminals and their ratio to all other terminals were significantly higher in the rostral portion of the circle of Willis and the proximal portion of its branches, than the caudal portion of the circle of Willis and the distal portion of its branches. Nerve terminals were observed to locate within 250 nm from the basal lamina of arterial smooth muscle cells in the rostral portion of the circle of Willis and proximal portion of its branching arteries. The present observation confirmed that NOS-containing nerve fibers truly innervate the smooth muscle cells of the arterial wall in the circle of Willis and its main branches. Close contact between NADPH-d-positive and -negative nerve fibers and terminals in these arterial portions may indicate that NOS-containing perivascular nerves may work to modulate the rest of the other perivascular nervous system, such as the sympathetic nerves, to regulate the homeostasis of the arterial tonus.

Retroperitoneal fibrosis and immune-complex glomerulonephritis.

A 63-year-old man with rapidly progressive glomerulonephritis of the immune-complex type showed typical findings of idiopathic retroperitoneal fibrosis involving the left ureter resulting in hydronephrosis. Treatment with steroid improved both conditions. Our case and previously reported cases showing the same association of conditions support the hypothesis that the association is not fortuitous but reflects a common immunological mechanism.

Stable methane hydrate above 2 GPa and the source of Titan's atmospheric methane.

Methane hydrate is thought to have been the dominant methane-containing phase in the nebula from which Saturn, Uranus, Neptune and their major moons formed. It accordingly plays an important role in formation models of Titan, Saturn's largest moon. Current understanding assumes that methane hydrate dissociates into ice and free methane in the pressure range 1-2 GPa (10-20 kbar), consistent with some theoretical and experimental studies. But such pressure-induced dissociation would have led to the early loss of methane from Titan's interior to its atmosphere, where it would rapidly have been destroyed by photochemical processes. This is difficult to reconcile with the observed presence of significant amounts of methane in Titan's present atmosphere. Here we report neutron and synchrotron X-ray diffraction studies that determine the thermodynamic behaviour of methane hydrate at pressures up to 10 GPa. We find structural transitions at about 1 and 2 GPa to new hydrate phases which remain stable to at least 10 GPa. This implies that the methane in the primordial core of Titan remained in stable hydrate phases throughout differentiation, eventually forming a layer of methane clathrate approximately 100 km thick within the ice mantle. This layer is a plausible source for the continuing replenishment of Titan's atmospheric methane.

Spiny keratoderma of the palms and soles - report of two cases.

We report two patients with spiny keratoderma of the palms and soles characterized by multiple tiny keratotic plugs on the palms and soles. This disease was reported to be possibly associated with internal malignancies. We found a tumor from the esophagus to cardia in one patient. Another had no tumor but the lesion occurred soon after a severe bronchial asthma attack. Causal relation between spiny keratoderma of the palms and soles and bronchial asthma is obscure. Since this disease has been under-diagnosed and under-reported, it is important for dermatologists to keep spiny keratoderma of the palms and soles in mind in daily clinical examinations.

Computer-aided generation of stimulation data and model identification for functional electrical stimulation (FES) control of lower extremities.

Standard stimulation data for unassisted standing up of paraplegic patients was generated by dynamic optimization linked with model simulation, to overcome the difficulties in the present electromyogram (EMG)-based method. The generated stimulation data were roughly in agreement with the normal subjects' EMG. From these, it is suggested that the 'model-based' method is useful as an alternative of the 'EMG-based method'. The same technique can be applied to generation of patient-specific stimulation data once the musculoskeletal system of a patient is properly identified. The musculoskeletal system must be identified from data taken from simple and noninvasive experiments for the identification method to be practically acceptable. We developed a musculoskeletal model and systematic identification protocols for this purpose. They were validated for the vastus lateralis muscle at the knee joint. The identification was successful and the predicted joint angle trajectories closely matched the experimental data. This implies that the model-based generation of patient-specific stimulation data is possible.

A case of symptomatic heterozygous female Fabry's disease without detectable mutation in the alpha-galactosidase gene.

We report a case of symptomatic heterozygous female Fabry's disease with low alpha-galactosidase blood activity. We could not find any mutations in the coding regions of either the signal peptide or the enzyme subunit in our case.

Effect of an antioxidant, ebselen, on development of chronic cerebral vasospasm after subarachnoid hemorrhage in primates.

BACKGROUND: Oxidation and/or free radical reactions after subarachnoid hemorrhage (SAH) may be involved in the development of chronic cerebral vasospasm. The inhibition of these reactions is thought to be one of the therapeutic strategies for prevention of cerebral vasospasm. We investigated the effect of Ebselen, a synthetic seleno-organic compound, which exhibits anti-oxidation by glutathione peroxidaselike activity to inhibit free radical reactions by lipid peroxidation on the development of chronic cerebral vasospasm in a primate model. METHODS: Seventeen monkeys were used. SAH was produced by introduction of a blood clot around the right middle cerebral artery and the right side of the circle of Willis in all animals. The monkeys were randomly divided into three groups according to Ebselen dosage: 1) no dosage or non-treated group; 2) high-dose Ebselen group; and 3) low-dose Ebselen group. The drug was administered at 10 mg/Kg in the high-dose group and 5 mg/Kg in the low-dose group twice a day in each group for 7 days after SAH. The vessel diameter was evaluated on angiograms before the induction of SAH and at Day 7 following SAH. RESULTS: In the untreated group, the angiograms showed significant (p < 0.05) reductions of the mean vessel caliber of the right internal carotid (ICA) (38 +/- 10% reduction) and the middle cerebral artery (MCA) (56 +/- 9.7%) compared with the baseline value before SAH. In the high-dose Ebselen-treated group, the mean percent reduction in vessel caliber of the right ICA (16 +/- 11%) and MCA (28 +/- 9.5%) on Day 7 angiograms were significantly (p < 0.05) lower than those in the nontreated group, whereas the mean percent reduction of these vessels in the low-dose Ebselen-treated group showed no significant difference compared with the untreated group. CONCLUSIONS: Chronic cerebral vasospasm was inhibited in the animals in which a relatively large amount of Ebselen was administered for 7 days after SAH. The results suggest that the oxidation or free radical reaction by lipid peroxidation after SAH might be involved in the pathogenesis of vasospasm, and that inhibition of these reactions by drugs, such as Ebselen, may have a promising effect for prevention of vasospasm.

Relationship between EMG signals and force in human vastus lateralis muscle using multiple bipolar wire electrodes.

This paper describes the relationship between knee extension force and EMG signals detected by multiple bipolar wire electrodes inserted into the human vastus lateralis muscle under isometric conditions. Six healthy male volunteers participated in this study. Eight pairs of bipolar wire electrodes were inserted into the right vastus lateralis muscle and the EMG data were simultaneously detected and analyzed. The EMG raw data and individual force-IEMG relations were influenced by the location of the electrode inserted into the muscle. The force and IEMG relationship averaged across subjects detected from the eight electrodes, however, showed almost the same linear correlation in spite of different electrode locations. No linear correlation was observed between MdF and the knee extension force. This result suggests that, if all of the muscle fibers participate in the same action at the same time, the averaged normalized IEMG from any places using wire electrodes could reflect the total activities of that muscle even if the muscle is large.

Localization of mRNAs for novel, atypical as well as conventional protein kinase C (PKC) isoforms in the brain of developing and mature rats.

Using in situ hybridization histochemistry, the localization of mRNAs for 10 isoforms of protein kinase C (PKC) in the rat brain was studied at embryonic and postnatal stages. In the embryonic brain, the gene expression was positive only for PKCepsilon, mu, lambda, and zeta with the former three more evident: The expression for PKCmu and lambda in the ventricular germinal zone and that for PKCepsilon, zeta, and lambda in the mantle zone. In the postnatal brain, the expression for PKCdelta, eta, and theta was detected differentially in a few circumscribed loci such as the thalamus, the habenula, the septum, and the cerebellar granule cells, whereas that for the other isoforms was seen widely in various loci of the gray matter with different intensity. The expression in the cerebellar external granule cell layer was positive only for PKCbeta (betaI and betaII), mu, and lambda with that for PKCbeta confined to its inner zone. There is a general tendency for all PKC isoforms that the expression levels reach at peaks in early postnatal brain and decreases more or less in adult specimens. This is the first report on the spatio-temporal heterogeneity in the gene expression for the whole members of PKC family in the brain throughout development, especially at embryonic days.