Ultrasound-guided gradual reduction using flexion and abduction continuous traction for developmental dysplasia of the hip: a new method of treatment.

We describe our experience in the reduction of dislocation of the hip secondary to developmental dysplasia using ultrasound-guided gradual reduction using flexion and abduction continuous traction (FACT-R). During a period of 13 years we treated 208 Suzuki type B or C complete dislocations of the hip in 202 children with a mean age of four months (0 to 11). The mean follow-up was 9.1 years (five to 16). The rate of reduction was 99.0%. There were no recurrent dislocations, and the rate of avascular necrosis of the femoral head was 1.0%. The rate of secondary surgery for residual acetabular dysplasia was 19.2%, and this was significantly higher in those children in whom the initial treatment was delayed or if other previous treatments had failed (p = 0.00045). The duration of FACT-R was significantly longer in severe dislocations (p = 0.001) or if previous treatments had failed (p = 0.018). This new method of treatment is effective and safe in these difficult cases and offers outcomes comparable to or better than those of standard methods.

Effect of clarithromycin on renal excretion of digoxin: interaction with P-glycoprotein.

We present a digoxin-clarithromycin interaction in two patients in whom digoxin concentrations were unexpectedly increased. The ratio of renal digoxin clearance to creatinine clearance in one patient was lower during the concomitant administration of clarithromycin (0.64 and 0.73) than that after cessation of clarithromycin administration (1.30 +/- 0.20; mean +/- SD). Because P-glycoprotein could play an important role in the renal secretion of digoxin, we hypothesized that clarithromycin decreases renal digoxin excretion by inhibiting P-glycoprotein-mediated transport. Digoxin transport was evaluated with use of a kidney epithelial cell line, which expresses the human P-glycoprotein on the apical membrane by transfection with MDR1 complementary deoxyribonucleic acid. Clarithromycin inhibited the transcellular transport of digoxin from the basolateral to the apical side in a concentration-dependent manner and concomitantly increased the cellular accumulation of digoxin. These results suggest that clarithromycin may inhibit the P-glycoprotein-mediated tubular secretion of digoxin, and this interaction mechanism may contribute to an increase in the serum digoxin concentration.

The morphology of single lateral vestibulospinal tract axons in the lower cervical spinal cord of the cat.

The intraspinal morphology of single lateral vestibulospinal tract (LVST) axons was investigated with the method of intra-axonal staining with horseradish peroxidase (HRP) and three-dimensional reconstruction of the axonal trajectory. Axons penetrated in the ventral funiculus at C5-C8 were identified as LVST axons by their monosynaptic responses to stimulation of the ipsilateral vestibular nerve and by their direct responses to stimulation of the ipsilateral Deiters' nucleus and LVST. Reconstructions were made from 34 well-stained LVST axons. Of these, 23 terminated in the brachial segments (C5-Th1) and the other 11 projected below Th2. These axons were traced over distances of 2.9-16.3 mm rostrocaudally. Within these lengths, one to seven axon collaterals (mean +/- S.D., 3.2 +/- 2.0, N = 19) were given off at right angles from the stem axons of LVST axons terminating in the brachial segments. The mean diameters of stem axons and primary collaterals were 4.5 microns and 1.6 micron, respectively. In the gray matter, collaterals ramified successively, pursued a delta-like path, and terminated mainly in laminae VII and VIII or lamina IX. The rostrocaudal extension of a single collateral was very restricted (mean +/- S.D., 760 +/- 220 microns, N = 16), in contrast to the extensive dorsoventral and mediolateral extent of the terminal arborization. There were usually gaps between adjacent collateral arborizations from the same stem axons, since the intercollateral distances ranged from 400 to 4,300 microns (mean = 1,490 microns). LVST axons terminating in brachial segments were divided into two groups--a medial group and a lateral group--on the basis of their projection sites in the transverse plane of the gray matter. The axons of the medial type had their main projection to laminae VII and VIII of Rexed, while those of the lateral type terminated in lamina IX. The terminal arborizations of the medial type LVST axons were mainly distributed over lamina VIII, where synaptic boutons appeared to make contact with proximal dendrites or somata of medium-sized and large neurons in the ventromedial nucleus and also in the medial portion of lamina VII adjacent to the central canal and dorsal to lamina VIII. Five out of 15 medial type axons had a bilateral projection. One or two collaterals of each of these axons crossed the midline through the anterior commissure and terminated in lamina VII or VIII. It was concluded that the contralateral projection was sparse.(ABSTRACT TRUNCATED AT 400 WORDS)

Morphology of single medial vestibulospinal tract axons in the upper cervical spinal cord of the cat.

The morphology of single medial vestibulospinal tract (MVST) axons was investigated by iontophoretic injection of horseradish peroxidase into single axons at the upper cervical cord in pentobarbital-anesthetized cats. MVST axons were identified by their monosynaptic responses to stimulation of the vestibular nerve and their direct responses to stimulation of the medial longitudinal fusciculus (MLF). Reconstructions of the axonal trajectory were made from 22 uncrossed and 19 crossed MVST axons at C1-C4. MVST axons ran in the ventral funiculus and gave rise to multiple axon collaterals to the upper cervical gray matter at different segments. These axons could be traced over the distance of 2.5-15.3 mm. Within these lengths, up to 9 axon collaterals were identified per axon (mean +/- s.d., 3.3 +/- 2.0, n = 41). Axon collaterals ramified in the gray matter several times and spread in a delta-like manner in both the transverse and horizontal planes. There were usually gaps free from terminal arborizations between adjacent axon collaterals, since the rostrocaudal extension of individual axon collaterals (mean = 820 microns) was very much limited in contrast to wide intercollateral intervals (mean = 1,510 microns). Axon terminals were distributed mainly in laminae IX, VIII, and VII, and sometimes in laminae VI-IV. Most abundant terminals were observed in lamina IX, including the ventromedial (VM), the spinal accessory (SA) nuclei and the nucleus dorsomedial to the VM nucleus (DM nucleus). A majority of individual axon collaterals provided some terminal branches to at least one of the above three motor nuclei. Axon collaterals projecting to laminae VIII-VI without terminals in the motor nuclei were rarely observed. Individual MVST axons had a preferential terminal distribution in each motor nucleus, but all three motor nuclei were covered by axon terminals of an ensemble of all MVST axons, indicating that all neck muscles innervated by these three motor nuclei are influenced by vestibular inputs through MVST axons. Most collaterals from a single axon produced circumscribed terminal arborizations in one or two common areas in the transverse plane (mainly in lamina IX) that were in line with one another in the longitudinal axis of the cord. This longitudinal arrangement of discontinuous terminal arborizations in lamina IX from a single axon may correspond to a continuous sagittal column of motoneurons for a particular muscle.(ABSTRACT TRUNCATED AT 400 WORDS)

Poly(A) dependent translation in rabbit reticulocyte lysate.

Poly(A) tail has been known to enhance mRNA translation in eukaryotic cells. However, the effect of poly(A) tail in in vitro is rather small. Rabbit reticulocyte lysate (RRL) is widely used for studying translation in vitro. Translation in RRL is typically performed in nuclease-treated lysate in which most of the endogenous mRNA have been removed. In this condition, the difference in the translational efficiency between poly(A)+ and poly(A)- mRNAs is about two-fold. We studied the effect of poly(A) tail on luciferase mRNA translation in nuclease untreated rabbit reticulocyte lysate, in which endogenous globin mRNAs were actively translated. In the case of capped mRNAs, stimulation of translation by poly(A) addition was about 1.5- to 1.6-fold and the effect of the poly(A) length was small. However, in the case of uncapped mRNAs, the addition of poly(A) tail increased luciferase expression over 10-fold. The effect of the poly(A) tail was dependent on its length. The difference in the translational efficiency was not due to the change of mRNA stability. These data indicate that RRL has the potential to translate mRNA in a poly(A) dependent manner.

Comparison of the branching patterns of lateral and medial vestibulospinal tract axons in the cervical spinal cord.

The morphology of single physiologically-identified lateral and medial vestibulospinal tract (LVST and MVST) axons was analysed, using intracellular staining with horseradish peroxidase (HRP) and three-dimensional reconstruction of axonal trajectories in the cat. Axons were penetrated in the cervical cord at C1-C8 with a microelectrode filled with 7% HRP. These axons were identified as vestibulospinal axons by their monosynaptic responses to stimulation of the vestibular nerve and further classified as either LVST or MVST axons by their responses to stimulation of the LVST and MVST. The stained axons could be traced over distances of 3-16 mm rostrocaudally. Within these lengths, both LVST and MVST axons were found to have multiple axon collaterals at different segments in the cervical cord. Up to seven collaterals were given off from the stems of MVST axons and LVST axons. The LVST axons included both neurones terminating at the cervical cord and those projecting further caudally to the thoracic or lumbar cord. Each collateral of these LVST axons, after entering into the gray matter, ramified successively in a delta-like fashion and terminated mainly in lamina VIII and in the medial part of lamina VII. Many boutons of both terminal and en passant types seemed to make contact with the cell bodies and proximal dendrites of neurones in the ventromedial nucleus (VM). Each collateral had a narrow rostrocaudal extension (0.2-1.6 mm, average 0.8 mm) in the gray matter in contrast to a much wider intercollateral interval (average 1.5 mm), so that there were gaps free from terminal boutons between adjacent collateral arborizations. The morphology of axon collaterals of MVST axons was very similar to that of LVST axons. The rostrocaudal extent of single axon collaterals was very restricted (0.3-2.1 mm) in contrast to the wide spread in a mediolateral or a dorsoventral direction. MVST axons had intensive projections to the upper cervical cord with multiple axon collaterals. One to seven collaterals of single MVST axons were found at C1-C3. Terminals of MVST axons were distributed in laminae VII, VIII and IX, including the VM, the nucleus spinalis n. accessorii (SA), and the commissural nucleus. Many terminals seemed to make contact with retrogradely-labelled motoneurones of neck muscles. Both axosomatic and axodendritic contacts were observed on motoneurones in various sizes. Some collaterals gave rise to terminal arborizations in both the VM and the SA. These results suggest that single LVST and MVST axons may control excitability of multiple dorsal axial muscles concurrently with their multiple axon collaterals at multisegmental levels.

Expression of substance P receptor in the substantia nigra.

Since the substantia nigra receives abundant substance P innervations but lacks clear evidences about a presence of substance P receptors, expressions for mRNA and protein of substance P receptors were investigated in the rat to resolve this mismatch. Expression levels of substance P receptors mRNA in the substantia nigra pars compacta and reticulata were 37.7 and 24.1% of those in the striatum, respectively, by reverse transcription-polymerase chain reaction (RT-PCR). Substance P receptors mRNA was found in dopamine neurons of the substantia nigra pars compacta by single cell RT-PCR. Ca. 90% of dopamine neurons in the substantia nigra pars compacta were immunoreactive to anti-substance P receptor antibody in the colchicine treated rats. These are the first direct evidence for the existence of substance P receptors in dopamine neurons of the substantia nigra pars compacta.

Synaptic organization of the vestibulo-collic pathways from six semicircular canals to motoneurons of different neck muscles.

The pattern of inputs from six semicircular canals to neck motoneurons was investigated by stimulating six ampullary nerves electrically and recording intracellular potentials from motoneurons of the rectus capitis dorsalis (RD), the complexus (COMP) and the obliquus capitis caudalis (OCA) muscles at the upper cervical cord of the cat. RD and COMP motoneurons received disynaptic excitation from bilateral anterior and contralateral horizontal ampullary nerves and disynaptic inhibition from bilateral posterior and ipsilateral horizontal ampullary nerves. OCA motoneurons received excitation from ipsilateral vertical and contralateral horizontal ampullary nerves and inhibition from contralateral vertical and ipsilateral horizontal ampullary nerves. Ipsilateral disynaptic inhibitory postsynaptic potentials and contralateral disynaptic excitatory postsynaptic potentials to these motoneurons were mediated by the medial longitudinal fasciculus (MLF) and the other postsynaptic potentials by the extra-MLF pathways. The results indicated that motoneurons of a neck muscle have its own characteristic pattern of inputs from six semicircular canals.

Comparison of carbohydrate-containing substances from non-calcified and calcified portions of bovine costal cartilage.

Carbohydrate-containing substances were extracted from non-calcified (NCC) and calcified (CC) portions of bovine costal cartilage with 0.5 M LaCl3 by the method of Mason and his co-workers, followed by dilution of the extract with 9 volumes of water. The precipitate formed on dilution yielded Fr. P, while Fr. S was obtained from the supernatant. Fr. P was separated into two subfractions by gel filtration on Sepharose 2B. The experimental results showed that Fr. P contained proteoglycans with different molecular sizes and compositions, while Fr. S contained proteoglycan, hyaluronic acid, glycoproteins, and glycogen. The present data suggest that in the proteoglycan of Fr. P, the relative content of chondroitin sulfate decreases with a concomitant increment in that of keratan sulfate on calcification. In addition, elevation of the ratio of chondroitin 4-sulfate to chondroitin 6-sulfate, together with a small increment of non-sulfated disaccharide units in the chondroitin sulfate chains appear to occur on calcification. The glycogen content in Fr. S diminished on calcification. The present observations suggest therefore that the remodeling of proteoglycan consumption of glycogen in bovine costal cartilage occur on calcification.

Four convergent patterns of input from the six semicircular canals to motoneurons of different neck muscles in the upper cervical cord.

This study was performed to investigate the pattern of input and the pathways from the six semicircular canals to motoneurons of various neck muscles in anesthetized cats. Intracellular postsynaptic potentials from neck motoneurons were recorded in response to electrical stimulation of the six ampullary nerves. The results showed that motoneurons of a particular neck muscle have a homogeneous convergent pattern of input from the six semicircular canals; there are four patterns of input from the six semicircular canals to motoneurons of various neck muscles; and the trisynaptic connection between the semicircular canal nerves and neck motoneurons was identified in addition to the disynaptic connection.

Projection pattern of single corticocortical fibers from the parietal cortex to the motor cortex.

Arborization of single corticocortical (CC) axons projecting from the parietal cortex to the motor cortex (Mx) was analysed using an intraaxonal staining technique in the cat. Stem axons arising from cell bodies in area 5 ramified repeatedly into numerous terminal branches in the Mx, forming 2-6 patches (0.2-0.8 mm in diameter) separated by a terminal-free gap. Axon terminals were distributed mainly in layers II and III and sparsely in layers V, VI and I. This feature is quite similar to that of thalamocortical axons and other corticocortical fibres. Thus the patchy organization may be a basic input structure for afferents of the Mx and play a role in generation of adequate motor output patterns in the Mx.

Synaptic organization of the cerebello-thalamo-cerebral pathway in the cat. I. Projection of individual cerebellar nuclei to single pyramidal tract neurons in areas 4 and 6.

The neural connections of the dentate (DN) and the interpositus (IN) nuclei to the motor cortex and area 6 were investigated by recording intracellular postsynaptic potentials from fast and slow pyramidal tract neurons (PTNs) in the anesthetized cat. Localized stimulation of DN and IN produced di- or polysynaptic EPSPs in fast and slow PTNs in the "forelimb area" of the motor cortex and area 6. The effects of stimulation of the two cerebellar projections were essentially the same, although some regional difference of their relative strength was noted. In these cortical areas, the majority of fast and slow PTNs received convergent inputs from both DN and IN. By examining the interaction of DN- and IN-evoked EPSPs, spatial facilitation and occlusion at the level of the thalamus were demonstrated. Therefore, it was concluded that at least a portion of the convergence of the dentate and the interpositus inputs occurred at the level of the ventrolateral nucleus of the thalamus.

Synaptic organization of the cerebello-thalamo-cerebral pathway in the cat. II. Input-output organization of single thalamocortical neurons in the ventrolateral thalamus.

Input-output neural organization of single thalamocortical (T-C) neurons in the ventrolateral nucleus (VL) of the thalamus was investigated using an intracellular recording technique in the anesthetized cat. Stimulation of the dentate (DN) and the interpositus (IN) nuclei produced monosynaptic unitary EPSPs of large amplitude in T-C neurons projecting to the motor cortex or area 6 over the entire mediolateral region of VL. The thalamic projections from DN and IN are very wide and there is a considerable overlap between the dentate and the interpositus projection areas in VL. And in this overlapping area, a considerable number of T-C neurons (50%) receive inputs from both DN and IN. More than 40% of T-C neurons were antidromically activated from widely separated electrodes in the motor cortex, indicating that the cortical arbolization of single T-C neurons is very wide and the number of these neurons with widely divergent projections is considerably large.

Synaptic organization of the cerebello-thalamo-cerebral pathway in the cat. III. Cerebellar input to corticofugal neurons destined for different subcortical nuclei in areas 4 and 6.

To analyze the cerebellar effects on corticofugal neurons destined for different subcortical nuclei, intracellular recordings were made from corticofugal neurons in areas 4 and 6 of the cat. Corticonuclear neurons to the red nucleus (RN) and the pontine nucleus (PN), and pyramidal tract neurons (PTNs) with collaterals to these nuclei were identified by their antidromic responses to the stimulation of these nuclei and the pyramid. Three types of RN-projecting neurons (corticorubral neurons (CRNs), corticopontine neurons (CPNs) with a collateral to the RN and PTNs with a collateral to the RN) and two types of PN-projecting neurons (CPNs and PTNs with a collateral to the PN) were differentiated. Furthermore, these corticofugal neurons were classified as fast and slow neurons on the basis of a critical axonal conduction velocity of 20 m/s. About 80% of 98 RN-projecting neurons in area 4 were PTNs, and among the rest, CPNs were more common than CRNs. A similar tendency of the frequency distribution of 37 RN-projecting neurons was also observed in area 6. In area 4, about 70% of 158 PN-projecting neurons were PTNs (80 fast and 30 slow PTNs) and the rest were CPNs, while in area 6, only 35% of 99 PN-projecting neurons were PTNs (10 fast and 25 slow PTNs). Among the CPNs in areas 4 and 6, slow CPNs were more frequently encountered. Cerebellar effects on these identified corticofugal neurons were investigated, using electrical stimulation of the brachium conjunctivum (BC). In both areas 4 and 6, a substantial number of fast conducting CRNs, CPNs and PTNs projecting to the RN or the PN received short-latency (predominantly disynaptic), large-amplitude EPSPs from the BC, and a considerable number of slow conducting neurons to the RN and/or the PN received longer-latency, smaller-amplitude EPSPs from the BC.

Glutamatergic and cholinergic inputs from the pedunculopontine tegmental nucleus to dopamine neurons in the substantia nigra pars compacta.

Postsynaptic responses of dopamine (DA) neurons in the substantia nigra pars compacta (SNc) to stimulation of the pedunculopontine tegmental nuclei (PPN) were studied in in vitro slice preparations in the rat. The recorded neurons were intracellularly injected with biocytin and also identified as DA neurons by an immunocytochemical technique. PPN stimulation induced in DA neurons monosynaptic excitatory postsynaptic potentials (EPSPs) that consisted of early transient and slow components. An application of anti-glutamatergic agents (1 mM kynurenic acid and/or 30 microM 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX)) in the bathing media partially suppressed the EPSPs, indicating that PPN inputs to SNc DA neurons are glutamatergic and non-glutamatergic. Anti-glutamatergic resistant EPSPs were suppressed by applications of anti-cholinergic agents such as atropine, mecamylamine, and pirenzepine. These data indicate a convergence of glutamatergic and cholinergic excitatory inputs from the PPN to SNc DA neurons and that both nicotinic and muscarinic receptors are involved in the cholinergic transmission.

Morphology of single neurones in the cerebello-rubrospinal system.

Axonal branching patterns of physiologically identified cerebellar nucleus neurones and rubrospinal neurones were investigated in the cat with intra-axonal injection of horseradish peroxidase and 3-dimensional reconstruction on serial sections. Axons of dentate and interpositus neurones projected to the VL nucleus of the thalamus and on their way, several axon collaterals were given off from the stem axons to the red nucleus. Axon terminals of interpositus neurones terminated as a sagittal sheet of arborizations in the red nucleus. Their terminal boutons made apparent contact with cell bodies and proximal dendrites of rubrospinal neurones. In rubrospinal axons, multiple axon collaterals were identified at different segments of the cervical spinal cord.

Convergent inputs from the dentate and the interpositus nuclei to pyramidal tract neurons in the motor cortex.

Effects of stimulation of the cerebellar nuclei were investigated by intracellular recordings from fast and slow pyramidal tract neurons and thalamocortical neurons in the cat. The present study demonstrated that: (1) the interpositus and the dentate nuclei excite PTNs in the motor cortex; (2) single pyramidal tract neurons receive convergent inputs from both nuclei, and (3) the convergence of the inputs from both nuclei occurs at the level of the ventrolateral nucleus of the thalamus.

Divergent projection of individual corticospinal axons to motoneurons of multiple muscles in the monkey.

Intracellular staining with horseradish peroxidase (HRP) of physiologically identified corticospinal (CS) axons originating from the monkey motor cortex revealed the intraspinal morphology of their branching patterns. CS collaterals spread in a delta-like fashion in the intermediate zone and lamina IX. Virtually all CS axons examined terminated in lamina IX, and it was shown by labeling motoneurons with retrograde transport of HRP that individual CS axons made direct contacts with dendrites of motoneurons of different muscle species.

Organization of excitatory inputs from the cerebral cortex to the cerebellar dentate nucleus.

Intracellular recording was made from dentate nucleus neurons (DNNs) in anesthetized cats, to investigate cerebral inputs to DNNs and their responsible pathways. Stimulation of the medial portion of the contralateral pericruciate cortex most effectively produced EPSPs followed by long-lasting IPSPs in DNNs. Stimulation of the pontine nucleus (PN), the nucleus reticularis tegmenti pontis (NRTP) and the inferior olive (IO) produced monosynaptic EPSPs and polysynaptic IPSPs in DNNs. The results indicate that the excitatory input from the cerebral cortex to DNNs is at least partly relayed via the PN, the NRTP and the IO. Intraaxonal injection of HRP visualized the morphology of mossy fibers from the PN to the DN and the cerebellar cortex. The functional significance of the excitatory inputs from the PN and the NRTP to the DN is discussed in relation to the motor control mechanisms of the cerebellum.