Supramolecular structure of the Salmonella typhimurium type III protein secretion system.

The type III secretion system of Salmonella typhimurium directs the translocation of proteins into host cells. Evolutionarily related to the flagellar assembly machinery, this system is also present in other pathogenic bacteria, but its organization is unknown. Electron microscopy revealed supramolecular structures spanning the inner and outer membranes of flagellated and nonflagellated strains; such structures were not detected in strains carrying null mutations in components of the type III apparatus. Isolated structures were found to contain at least three proteins of this secretion system. Thus, the type III apparatus of S. typhimurium, and presumably other bacteria, exists as a supramolecular structure in the bacterial envelope.

A branched pathway in the early stage of transcription by Escherichia coli RNA polymerase.

The time-course of synthesis of long and short (abortive) transcripts by Escherichia coli RNA polymerase was investigated under single round conditions in vitro. The synthesis of long RNA initiated at the lambda PR (with an altered leader sequence) or at the lacUV5 promoter was completed within five minutes, but abortive transcripts were continuously synthesized for at least 20 minutes. The results indicate the presence of transcription complexes that are capable only of abortive synthesis, and not of productive elongation. Consistently, only one in four of the RNA polymerase molecules that initially associated with the lambda PR promoter synthesized long RNA. The enzyme reisolated from productive complexes synthesized both long and abortive transcripts, behaving just like the original enzyme. This suggests that RNA polymerase is homogeneous with respect to its ability to synthesize the two types of product. Overall, the results indicate that many transcription complexes can be irreversibly trapped in vitro in an abortive cycle.

Morphological pathway of flagellar assembly in Salmonella typhimurium.

The process of flagellar assembly was investigated in Salmonella typhimurium. Seven types of flagellar precursors produced by various flagellar mutants were purified by CsCl density gradient protocol. They were characterized morphologically by electron microscopy, and biochemically by two-dimensional gel electrophoresis. The MS ring is formed in the absence of any other flagellar components, including the switch complex and the putative export apparatus. Four proteins previously identified as rod components, FlgB, FlgC, FlgF, FlgG, and another protein, FliE, assemble co-operatively into a stable structure. The hook is formed in two distinct steps; formation of its proximal part and elongation. Proximal part formation occurs, but elongation does not occur, in the absence of the LP ring. FlgD is necessary for hook formation, but not for LP-ring formation. A revised pathway of flagellar assembly is proposed based on these and other results.

Abnormal contingent negative variation in writer's cramp.

OBJECTIVE: To investigate the physiological abnormality in writer's cramp, a focal dystonia which specifically affects writing. METHODS: We recorded brain potentials that precede hand and neck movements (contingent negative variation or CNV) in 11 patients and 11 age-matched normal subjects. A 1000 Hz tone burst (S1) was delivered to the right or left ear in random sequence, and 2 s after, a 2000 Hz tone burst (S2) was delivered to both ears simultaneously. For the response task to S2, the subjects were instructed to extend their fingers ipsilateral to the ear to which S1 was given in one experiment or to rotate the head to the side of the S1 presentation in another. All the patients had symptoms in the right hand only, and performed both tasks normally. CNV amplitudes were compared between normals and patients using unpaired t test. RESULTS: They showed normal CNV for neck movement but significantly decreased CNV amplitudes for movements both in the affected and unaffected hands. CONCLUSIONS: Our findings suggest that motor programming is specifically abnormal for the affected body part, including the asymptomatic contralateral limb, and that the clinical symptom may result from a deficient compensatory mechanism for abnormal motor programs or subroutines.

[Treatment of spasticity with botulinum toxin and muscle afferent block].

Spasticity is partly caused by disinhibition of spinal stretch reflex activity and blocking of the group Ia afferents may improve its clinical symptoms. Since muscle afferent block has been successfully used for treating dystonia, this method may become useful for treating spasticity as well. We injected diluted lidocaine (0.5%, 50 ml/session) and ethanol (5 ml) into thigh adductors (for leg crossing), medial hamstrings (for medial rotation of the leg), quadriceps femoris (for hyperextension of the knee), tibialis posterior and triceps surae muscles (pes equinovarus deformity), biceps brachii and forearm flexors (for flexion deformities of the upper extremity) in 12 patients with spasticity. Although the duration of action was short (< 1 day) at first, it was gradually prolonged to several weeks by repeating the injection every 3-4 days. The final outcome was comparable to that obtained by botulinum toxin injection in the same group of patients. This method may prove its promise as a means of treating spasticity.

[A case of acute motor sensory axonal polyneuropathy after Haemophilus influenzae infection].

A 47-year-old woman developed consciousness disturbance, and experienced hallucinations while traveling abroad, and then went into critical condition. She was placed in the critical care unit, and had flaccid tetraparesis requiring mechanical ventilation. Haemophilus influenzae was cultured from the sputum. The level of protein of the cerebrospinal fluid was elevated to 114 mg/dl, nerve conduction study showed findings of pure axonal damage, and the sural nerve biopsy revealed severe axonal degeneration. She improved gradually by plasma exchange. The diagnosis of acute motor sensory axonal polyneuropathy (AMSAN) based on autoimmune mechanism was made. We speculate that H. influenzae infection may have elicited AMSAN in this case.

[Sequential studies of motor nerve conduction in relation to the disease stages of Guillain-Barré syndrome].

We retrospectively studied sequential changes in motor nerve conduction in forearm segments of 17 median nerves in 9 consecutive patients with Guillain-Barré syndrome (GBS). The disease course was divided into 6 stages: progressing, nadir, early-, mid- and late-convalescent, and cured stages. All patients were tested on at least 3 occasions in each different stage. Low amplitude compound muscle action potentials (CMAP) were noted in the progressing and nadir stages, suggesting the presence of conduction block. In the early- and mid-convalescent stages, on the other hand, the delays in distal motor latencies and conduction velocities as well as temporal dispersions of CMAPs were more prominent than in the progressing or nadir stage. These findings suggest that pathophysiological processes are different in the two stages of GBS: demyelination with conduction block predominated in the earlier stage and demyelination without conduction block predominated in the later stage. The former is considered to involve predominantly the relatively smaller motor nerves during the progressing and nadir stages, producing muscle weakness but not causing marked slowing of the motor nerve conduction. The latter is considered to involve predominantly the fastest motor fibers in the later days lasting into the early- and even mid-convalescent stages. This process seems to produce marked conduction slowing, but does not cause prominent muscle weakness. In addition, recovery from the conduction block during the convalescent stages produces temporal dispersions of CMAPs because of the variably slowed conduction velocities.(ABSTRACT TRUNCATED AT 250 WORDS)

[Muscle afferent block for the treatment of writer's cramp].

A 29-year-old man suffered from dystonic writer's cramp for over three years. When he wrote, typed and did other tasks using right hand, dystonic involuntary movement triggered medial rotation of the arm, wrist extension and shoulder elevation. Medication, biofeedback, and botulinum injection were performed without much success. We tried to block the sensory input from muscles by using lidocaine and ethanol. We made injections of 0.5% lidocaine 50ml and 99% ethanol 5ml into muscles with abnormal activity at the frequency of twice a week for about six months. After the treatment, dystonic movement was remarkably improved and he was then able to write, type and perform other tasks with the right hand. Side effects included pain of the injection site, nausea and dizziness, which lasted for a few hours. This "muscle afferent block" did not cause muscle weakness. We speculate that muscle afferent plays a pivotal role in dystonia so that its blocking may be of clinical use.

[Cerebral infarction presenting pure motor monoparesis: diagnosis by diffusion-weighted MR imaging].

We studied 10 patients with acute ischemic cerebrovascular disorders presenting paralysis confined to one limb, unaccompanied by sensory signs(pure motor monoparesis, PMM) on diffusion-weighted MR imaging(DWI). DWI revealed fresh ischemic lesions in all patients, except for 2 cases of transient ischemic attack. On DWI, acute infarction in multiple lesions was identified, and small superficial lesions were clearly described. Superficial lesions were seen in 4 patients, and deep lesions were also seen in 4 patients. DWI is useful for lesion analysis in cerebral infarction with PMM.

[A juvenile case of chronic inflammatory demyelinating polyradiculoneuropathy with severe onion bulb-like change mimicking hereditary neuropathy].

A 15-year-old male developed symmetrical weakness of the limb muscles. He had not had any previous developmental disorders except delayed initiation of walking. Flexion contraction of ankle joint and pes cavus deformity were seen. The cerebrospinal fluid protein concentration was elevated. Nerve conduction study showed severe conduction block and temporal dispersion. A sural nerve biopsy revealed remarkable onion bulb-like changes and perivascular infiltration of inflammatory cells. After high-dose corticosteroid treatment, he showed improvement in muscle strength. Although there were no abnormalities of genes related to hereditary neuropathy, the atypical findings of severe demyelinating changes of peripheral nerves mimicked hereditary neuropathy.

[The clinical usefulness of high-dose intravenous immunoglobulin therapy for chronic inflammatory demyelinating polyneuropathy and multifocal motor neuropathy].

To explore the optimum dose of intravenous immunoglobulin (i.v.Ig) for treating patients with chronic inflammatory demyelinating polyrneuropathy and multifocal motor neuropathy, we compared the usefulness of i.v.Ig among 3 treatment doses. Fifty-nine patients were randomly divided into three treatment dosage groups: 20 patients for Group I using 50 mg/kg/day x 5 days, 19 patients Group II using 200 mg/kg/day x 5 days, and 20 patients Group III using 400 mg/kg/day x 5 days. We assessed clinically and electrophysiologically the effectiveness of the treatment at 5 weeks after the initial infusion. For patients in Group I and II who had not improved (or worsened) with the first treatment, we gave a one-step larger dose in the second treatment (i.e. 200 mg/kg/day x 5 days for those who had been given 50 mg/kg/day x 5 days, 400 mg/kg/day x 5 days for those who had been given 200 mg/kg/day x 5 days) after more than 9 weeks. We found that 15% of the patients in Group I, 21% in Group II and 60% in Group III improved dose-dependently with the first intravenous immunoglobulin treatment. Seven (47%) of 16 patients in Group I and 4 (40%) of 11 patients in Group II improved after the second treatment with larger doses. Adverse reactions including chill sensation, fever, skin eruption and increase in blood GOT and GPT levels were transient and mild. One patient in Group III developed left hemiparesis showing the small infarction in the right thalamus during the course of the treatment, but the symptom was mild. In conclusion, the high-dose intravenous immunoglobulin therapy (400 mg/kg/day x 5 days) is useful for treating patients with CIDP and MMN, although care must be taken of the risk of causing cerebral infarctions.

Dementia in Parkinson's disease: diffusion tensor imaging.

OBJECTIVE: Dementia occurs frequently in patients with Parkinson's disease (PD). However, the nature of the dementing process remains controversial. We evaluated various cognitive functions in patients with PD, compared fractional anisotropy (FA) values between PD patients with and without dementia. METHODS: Thirty-seven consecutive patients with Hoehn-Yahr stage III or IV PD participated in this study. Patients were divided into two groups: (i) PD with dementia group (PDD) and (ii) PD without dementia group (PDND). There were 11 PDD and 26 PDND cases. Ten controls were also studied. RESULTS: The PDD group showed significant FA reduction in the bilateral posterior cingulate bundles compared with PDND. FA values in the left posterior cingulate bundle showed significant correlations with many cognitive parameters. INTERPRETATION: Our results showed that the posterior cingulate areas play some important roles in the dementing process in PDD. However, as the pathological processes responsible for dementia in PD patients may be multifaceted, further studies are necessary.

Wisconsin Card Sorting Test in Parkinson's disease: diffusion tensor imaging.

INTRODUCTION: It is generally assumed that executive dysfunctions in Parkinson's disease (PD) are caused by degeneration of the basal ganglia or frontal cortex or both. However, there have been few studies investigating the relationship between executive dysfunctions and cerebral pathological change. The objective of this study was to evaluate various cognitive functions in non-demented patients with PD, and to compare the fractional anisotropy (FA) values of PD patients with and without executive dysfunction. MATERIALS AND METHODS: Twenty-one consecutive non-demented patients with PD were enrolled in this study. Patients were divided into two groups on the basis of their Wisconsin Card Sorting Test score. RESULTS: There was significant FA reduction in the left parietal white matter in the group in which the number of categories achieved was 2. CONCLUSION: Accumulating evidence suggests that conventional 'frontal' tasks correlate with both frontal lobe and parietal lobe function, and we suggest that pathological changes in the left parietal lobe may cause, in part, disturbances in executive tasks in PD.

Three-dimensional stereotactic surface projection study of orthostatic hypotension and brain perfusion image in Parkinson's disease.

OBJECTIVE: To compare brain perfusion image using three-dimensional stereotactic surface projection (3D-SSP) analysis of N-isopropyl-p-123I iodoamphetamine (123I-IMP) single photon emission computed tomography (SPECT) between patients with Parkinson's disease with orthostatic hypotension and those without orthostatic hypotension. MATERIALS AND METHODS: Fifteen patients with Parkinson's disease and orthostatic hypotension and 13 patients with Parkinson's disease without orthostatic hypotension were studied. We compared brain perfusion image between the two groups by 3D-SSP. RESULTS: Bilateral anterior cingulate gyrus perfusion of the patients with orthostatic hypotension was significantly decreased compared to that of the patients without orthostatic hypotension. CONCLUSIONS: The disorder of anterior cingulate gyrus may participate in the autonomic failure in Parkinson's disease.

CD16+CD57- natural killer cells in multifocal motor neuropathy.

We analyzed the CD16+CD57- lymphocyte subset, which is considered to have strong natural killer (NK) cell activity, in peripheral blood from patients with chronic immune-mediated neuropathies and patients with other neurological diseases. We found that the ratio of CD16+CD57- NK cells to total lymphocytes was increased in 4 of 6 patients with multifocal motor neuropathy (MMN) with persistent conduction block. Since the CD16 molecule is an Fc receptor for immunoglobulin G (IgG), high-dose intravenous immunoglobulin (IVIg) may interfere with CD16+CD57- NK cells via Fc receptor blockade. In addition, cyclophosphamide (Cy) is often used to suppress NK cells. Therefore, our findings may partly account for the effectiveness of IVIg or Cy, which is the current treatment of choice for MMN.

Physical interference between escherichia coli RNA polymerase molecules transcribing in tandem enhances abortive synthesis and misincorporation.

Transcription initiation is accompanied with iterative synthesis and release of short transcripts. The molar ratio of enzyme to template was found to be critical for the amounts and distribution of the abortive products synthesized by Escherichia coli RNA polymerase from several promoters. At a high ratio abortive synthesis of 4-8 nt were enhanced at thelambda P R promoter. Removing excess RNA polymerase just before initiation, achieved by washing immobilized transcription complexes, prevented this enhancement. At this high ratio synthesis of an unexpected 6 nt transcript was enhanced when the enzyme stalled at position +32, but not when it stalled at position +73. This transcript had misincorporations at its fifth and sixth positions, probably due to slippage. Hydroxyl radical footprinting of the complex stalled at +32 in the presence of excess enzyme showed that more than one molecule of RNA polymerase was tandemly bound to the same DNA. These results suggest that: (i) when RNA polymerase molecules are tandemly transcribing the same DNA, transient collisions enhance abortive synthesis by the trailing molecule; (ii) when the leading polymerase stalled in the initially transcribed region blocks progression of the trailing polymerase, the latter can commit misincorporations, probably due to slippage synthesis.

Kinetics of transcription in a minute column.

Immobilized enzymes or nucleic acids are widely used to analyze association and dissociation reactions. A pseudo-rapid kinetic method for studying transcription was developed by using an immobilized template DNA packed in a minute column, in a micro-scale liquid chromatography system. This method, which has a dead-time of only several seconds, is quick enough to allow analysis of the release of product RNA in transcription by Escherichia coli RNA polymerase. The method could be applied to most enzymes that interact with DNA.