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Maize waxy (Wx) protein, which has been detected as a single protein with a M r 60 000 by sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE), was separated into four isoforms by 2D-PAGE. The isoelectric point (p I ) of these isoforms ranged from 6.0 to 6.5. Starch isolation with SDS-buffer and heat-extraction of protein from starch granules had no effect on the 2D electrophoretic patterns of Wx protein. Rice and barley Wx proteins were also analysed by the same method and separated into four and three isoforms, respectively. Rice Wx protein isoforms had a M r of 60 000 as had been reported with a p I range from 6.5 to 7.2 while barley Wx protein isoforms had a M r of 61 000 and p I in the range of 5.8 to 6.3.

A 57-year-old woman presented with weakness of the right leg over one and a half years. She had visited several clinics but her symptom had been attributed to lumbar spondylosis or aging itself. Motor examination revealed 4/5 strength in the distal right lower limb without muscle atrophy. Hyperreflexia of the right upper and lower limbs with right-sided ankle pseudoclonus, right positive Babinski’s sign and sensory impairment in her right leg below the knee were also noted. Brain magnetic resonance imaging revealed a left parasagittal parietal meningioma with homogeneous contrast enhancement (Picture). Monoparesis of the leg is commonly caused by a lower motor neuron lesion; however, it could be attributable to a central lesion such as meningioma (1, 2). Here, we report a rare case of pure motor monoparesis of the leg due to parasagittal meningioma and emphasize that detecting upper motor neuron signs such as hyperreflexia, clonus or positive Babinski’s sign in an affected limb is crucial to reach the correct diagnosis of localization.

A 24-year-old woman presented subdural empyema developing from sinusitis caused by Porphyromonas and Fusobacterium infection. She experienced fever and progressive headache with gradually worsening symptoms. Neurological examination revealed drowsiness and neck stiffness. A CSF examination detected pleocytosis and a low glucose level. Gadolinium-enhanced T1-weighted images from brain MRI showed thickening enhancement at the leptomeninges in the left frontal to temporal lobes and interhemispheric fissure with edema. Based on the diagnosis of bacterial meningoencephalitis and subdural empyema developing from sinusitis primary to odontogenic infection, she received antibacterial chemotherapy with meropenem hydrate and vancomycin hydrochloride, after which motor aphasia and consciousness disturbance occurred. No bacteria were isolated from a trans-sphenoidal biopsy specimen and CSF culture. Molecular typing also was performed by sequencing the 16S ribosomal RNA intergenic spacer region, and Porphyromonas and Fusobacterium were identified. She was given cephalosporin and metronidazol, after which her neurological symptoms and signs gradually lessened. Physicians need to be aware that patients may develop subdural empyema subsequent to sinusitis associated with Porphyromonas and Fusobacterium infection and that amplification and sequence analysis of partial bacterial 16S ribosomal RNA gene should be examined when no bacteria is identified by culture.

We report the case of a 50-year-old man with paraneoplastic cerebellar degeneration (PCD) and Lambert-Eaton myasthenic syndrome (LEMS) associated with primary double lung cancer. He developed acute progressive double vision, slurred speech, and gait disturbance. Neurological examination revealed diplopia, mild ptosis, bilateral horizontal gaze-evoked nystagmus, and cerebellar limb and truncal ataxia. The diffusion image of brain magnetic resonance imaging (MRI) revealed no abnormal findings in the cerebellum. On the basis of the diagnosis of acute cerebelitis, he was given methylprednisolone pulse therapy followed by oral prednisolone, which gradually improved his neurological signs and symptoms. The analysis of the possible etiology suggested that the PCD was induced by lung cancer, which led to ataxia. A chest computed tomography scan revealed mass lesions of irregular shape and unclear margins in the upper lobe of the right lung and a small nodule tumor in the upper lobe of the left lung. We performed transbronchial needle aspiration and detected the bronchioloalveolar carcinoma of the right lung. An electromyogram showed waxing phenomenon in the ulnar nerve at high-frequency (50Hz) stimulation. The serum levels of anti-P/Q-type voltage-gated calcium channel (VGCC) antibody were elavated in the patient. These findings confirmed that the pathogenesis of the condition of this patient to be associated with LEMS. His cerebellar symptoms were considered to be caused by the PCD, and the diplopia, ptosis, and hyporeflexia were attributed to LEMS. We performed upper left lobectomy with mediastinal lymphnode dissection via video-assisted thoracoscopic surgery. A histological study detected small cell carcinoma. A diagnosis of double primary lung cancer was made. Physicians need to be aware that patients may develop PCD and LEMS associated with anti-VGCC antibody caused by small cell lung cancer, and a mass survey should be conducted and careful examinations performed.

The Japanese rice cultivar Asominori shows high bacterial leaf blight resistance (BLBR). The resistance locus is closely linked with the Ph gene on chromosome 4. Japanese rice breeders noticed that lines resistant to both bacterial leaf blight (BLB) and rice leaf blast (LB) segregated from descendants of crosses with Asominori; however, the cause of this cosegregation was unclear. In 2004, we obtained five Asominori backcrossed inbred lines (BILs). Firstly, we evaluated the resistance to LB and BLB in these BILs. The partial resistance to LB was high, similar to criterial varieties such as Ouu320 and Chubu45. The BILs were also resistant to the BLB strain T-7174 (race1). Using 21 PCR-based DNA markers, we analyzed the genotypes of the BILs on chromosome 4, where BLBR and Ph genes are located. All five BILs carried a common Asominori-derived chromosomal segment estimated to be 1336 kbp long. Secondly, segregation of BLBR and LBR was monitored using 118 F3 lines obtained from a cross between Ukei854 and Hitomebore, a BLB- and LB-susceptible cultivar. Most resulting F3 lines showing BLBR also carried LBR; however, several lines showed only BLBR or BLR, indicating that BLBR and LBR were controlled by very closely linked loci. Using RM5473-2, we surveyed the genotypes of the 118 F3 lines located on the common segment. Most F3 lines with a high LBR showed genotypes identical to Asominori. Thus, the LBR locus in Asominori was tightly linked to the BLBR locus on chromosome 4, and the LBR gene was probably located near RM5473-2.

The Cape Ohma region of Shimokita Peninsula, the northernmost point of Honshu Island, Japan, is subject to both the warm Tsugaru Current and the cold Kurile Current. As a result, the Laminarialean flora includes both cold temperature species (Laminaria japonica Areschoug, Kjellmaniella crassifolia Miyabe and Costaria costata (C. Agardh) Saunders) andwarm temperature species (Undaria peterseniana (Kjellman) Okamura, Ecklonia stolonifera Okamura), as well as Undaria pinnatifida (Yendo) Okamura, which is distributed in both waters. The frequency of occurrence (as a measure of distribution) and the biomass of these species were recorded in June 1976 (at 50 points in depths between 8–30 m), July 1988 (192 points, 2.5–25 m) and July 2001 (78 points, 2.5–25 m). Comparison of these data revealed a decrease in cold temperature species and an increase in warm temperature species from 1976 or 1988 to 2001. Long-term data of seawater temperature measured at 5 m depth near the study site showed that mean temperatures in the middle of winter (late January to February) in 1989–2000 were 0.9–1.1 °C higher than those in 1980-1988. Higher seawater temperatures in the last decade appear to have affected the frequency of occurrence and biomass of the Laminarialean species along the coasts of Cape Ohma. This result supported our previous conclusion that 1 °C higher mean seawater temperature in late January caused a decrease in the biomass of L. japonica (by ca. 64%) along the same coast.

As the result of developments in computer technology in recent years, the calculation, simulation and analysis of the stress environment for the artificial hip prosthesis and the femur have been made possible. In order to perform an exact calculation, three-dimensional modeling with the hip prosthesis and femur is required. The modeling of a femur is done by reconstructing the femoral outline data from CT images using three-dimensional CAD. The outline extraction work is performed by the experience and subjectivity of the operator, and the reliability of the data and working efficiency is low. In this paper, algorithms that extract the femur outline from CT images automatically are proposed.

The major components of storage starch are amylose and amylopectin, and in wheat, both an amylose-free mutant lacking granule-bound starch synthase I and a high-amylose mutant lacking starch synthase IIa have been produced recently. Here, we report the production of an amylose-free/ high-amylose double mutant. This double mutant has kernel and carbohydrate characteristics that are remarkably different than those of either single mutant, including a dramatically shrunken seed shape. Surprisingly, the double mutant has maltose and sucrose levels that are high enough to make it worthy of being called "sweet wheat".

A number of cold responsive (Cor)/late embryogenesis abundant (Lea) genes are induced by both low temperature (LT) and dehydration. To understand the molecular basis of cold acclimation and its relationship with drought stress response in wheat seedlings, we isolated a DREB2 homolog Wdreb2, which is the candidate gene for a transcription factor of the Cor/Lea genes. The Wdreb2 expression was activated by cold, drought, salt and exogenous ABA treatment. Detailed expression studies of Wdreb2 indicated the involvement of two distinct pathways of its activation, a drought and salt stress-responsive pathway and a cold-responsive pathway. The transient expression analysis showed that the Wrab19 expression was directly activated by the WDREB2 transcription factor in wheat cells. Three transcript forms of Wdreb2 (Wdreb2alpha, Wdreb2beta and Wdreb2gamma) were produced through alternative splicing. Under drought and salt stress conditions, the amount of the Wdreb2beta form remained fairly constant during 24-hour treatment, while those of the Wdreb2alpha and Wdreb2gamma forms showed transient increases. On the other hand, the LT treatment resulted in increased transcript levels of all three forms of Wdreb2. Thus, under the LT and drought/salt stress conditions the amount of the WDREB2 transcription factors in wheat is differentially controlled by the level of transcription and alternative splicing.

PCR selection markers for the identification of null waxy alleles were used to screen for waxy mutations in 168 common wheat cultivars. In all cultivars where the Wx-B1 protein was absent, the Wx-B1 allele was identical to the previously identified mutation carried by Kanto 107. Although most cultivars missing the Wx-A1 protein also carried the same Wx-A1 mutation as found in Kanto 107, all of the Turkey Wx-A1 mutants produced a different PCR fragment, implying the presence of a different mutation. Sequencing of this fragment indicated the mutation, which consisted of a 173-bp insertion in an exon, was in a different location than the previously identified Wx-A1 mutation. An 8-bp duplication of the Wx-A1 sequence flanked each end of the insertion, and an element with reverse complementary sequences was present at both ends of the insertion. These structures correspond with the features of class II transposable elements. Hence, the Turkey null Wx-A1 mutation was likely caused by the movement of a transposon, and this spontaneous mutation appears to be present in a limited geographical area.

Task-specific dystonia (TSD) is a disabling movement disorder characterized by focal, involuntary, and excessive muscle contractions that develop in a single body part involved in highly skilled tasks [1]. In TSD, involvement of the hand (while writing, typing, or playing a musical instrument) is more common than the involvement of other parts of the body, but TSD related to the use of a computer mouse has never been described. A 68-year-old right-handed man presented to our outpatient clinic with involuntary flexion of his right thumb, which interfered with the use of a computer mouse. When the patient was 62 years old, he started composing music using a computer. Since then, he had used the computer mouse with his right hand for composition for approximately 5 h per day. At age 64, he developed abnormal flexion of his right thumb whenever he used the computer mouse; the flexion interfered with moving the mouse, but he could click the mouse button without any trouble. During the following year, abnormal flexion in the right thumb was observed even when the patient put his right hand on the table, and he had difficulty in extending and flexing his right thumb voluntarily. This forced him to use the computer mouse with his left hand instead. He had no symptoms in other parts of the body. The patient had played classical guitar since childhood. He had no previous medical history of psychiatric disease or a family history of dystonia. The patient had no history of alcohol use or smoking. The initial examination showed excessive flexion of his right thumb, and voluntary flexion and extension of his right thumb was impaired. This abnormal posture significantly worsened when using the computer mouse (see video, segment 1). However, he noticed that right-thumb flexion never occurred while eating (using chopsticks, spoon, or knife) or writing with a pen. The abnormal flexion of his right thumb never occurred when his right thumb and index finger touched. A neurological examination was unremarkable except for the abnormal flexion position of his right thumb. Laboratory data were normal. The finding of brain magnetic resonance images was normal. TSD was diagnosed based on the observation that involuntary and excessive flexion posture was prominent in specific situations. Oral medications such as levodopa, carbamazepine, and phenytoin were ineffective. The patient was then treated with clonazepam 0.5 mg, which modestly improved his symptoms. The combined use of clonazepam 1 mg and trihexyphenidyl 6 mg resulted in significant improvement of his symptoms (see video, segment 2). We report the first case of a patient in whom excessive flexion of the right thumb occurred initially only when using the computer mouse. Similar to other TSD patients [1], our patient’s symptoms later involved other tasks. As the patient had dystonic posturing of the right hand at rest at the examination, the patient was classified as having complex TSD rather than simple TSD [2]. His finger movements were preserved when his right thumb and index Electronic supplementary material The online version of this article (doi:10.1007/s00415-012-6519-1) contains supplementary material, which is available to authorized users.

Mutations in the three homeologous waxy loci Wx-A1, Wx-B1, and Wx-D1 of a waxy wheat line have previously been characterized at the molecular level. Using combinations of these mutations, six types of partial waxy wheat plus wild type and waxy wheat (types 1–8) can be produced. Here, we describe primer sets for all three loci that can be used under a single set of conditions, allowing 32 lines to be characterized as types 1–8 in a single PCR run using a 96-well plate. Using multiplex PCR, mutations at the Wx-B1 and Wx-D1 loci can be identified in a single PCR, reducing the number of reactions necessary to identify and select the desired partial waxy wheat line. A single multiplex PCR can be used to detect all three mutations when products are analyzed using capillary electrophoresis on a microchip device. The PCR conditions and primers are effective with a number of cultivars from other countries, indicating that the mutations found at the Wx-A1 and Wx-B1 loci of these cultivars likely have the same origins as the mutations in the corresponding loci of the waxy wheat line used in this study. The PCR selection method described here is an easy and effective alternative to the commonly used SDS–PAGE methods for identification of null alleles.Key words: partial waxy wheat, amylose, PCR, capillary electrophoresis, marker-assisted selection.

Dear Sirs, Food intake is regulated by homeostatic and reward-related pathways. The hypothalamus and brainstem integrate homeostatic signals with reward-related pathways, such as the dopaminergic neurons of the ventral tegmental area, nucleus accumbens, insular cortex, anterior cingulate cortex, and orbitofrontal cortex [1]. Eating disorders following brain lesions of the frontal and temporal lobe, hypothalamus, thalamus, and brainstem have all been described [2–4]. A 58-year-old right-handed woman suddenly presented with abnormal eating behavior and excessive daytime sleepiness in August 2009. The patient was seen consuming a large amount of salt-boiled, green edamame-type soybeans by a family member, and she slept throughout the day, except for when she was eating, on the day before her admission. The patient had no history of eating disorder. Neurological examination was unremarkable except for memory disturbance. She scored 26 points on the MiniMental State Examination (MMSE) scale (the impaired items being related to orientation and recall). No frontal release signs were observed. Brain magnetic resonance imaging (MRI) showed an acute ischemic lesion in the right capsular genu (Fig. 1a), with deep white matter lesions. Sixteen days after onset of symptoms, the patient was discharged with mild residual cognitive impairment and daytime sleepiness. After discharge, the patient could not resist the urge to eat and consumed three packs of edamame per day, resulting in weight gain of 5 kg over 3 months. The feeling of hunger was not necessary for this eating behavior to occur. The patient admitted that, before the onset of stroke, she had wanted to eat edamame whenever she saw her husband eating them while drinking beer but that she had previously restrained herself from doing so. She did not display increased eating behavior with other foods. Her appetite and eating behavior returned to normal by November 2009. In August 2010, she returned to our outpatient clinic with a 2-month history of the previously observed symptoms of daytime sleepiness and increased consumption of edamame. Brain MRI showed bilateral capsular genu infarcts (Fig. 1b). Brain single-photon emission computed tomography (SPECT) showed hypoperfusion in the bilateral frontal lobes (Fig. 1c, d). She scored 23 points on the MMSE scale (the impaired items being related to orientation, attention, calculation, and recall). We report a patient who developed right and left capsular genu infarcts on two separate occasions; both episodes were associated with identical clinical symptoms of increased reward-related feeding and daytime sleepiness. The patient displayed an irrational desire to consume edamame. Edamame is a nutritious green vegetable, also known as fresh green soybean, which is harvested at the peak of ripening just before it reaches a hardening stage. The stroke of each side of the capsular genu in the same patient on different occasions, which induced reproducible abnormal eating behavior related only to palatable food in the absence of frontal release signs, suggests the involvement of reward-related food pathways. This behavior is in contrast with binge eating, which is observed in patients with thalamic infarcts [2, 4]. However, because our patient K. Suzuki (&) T. Sada H. Takekawa K. Hirata Department of Neurology, Dokkyo Medical University, 880 Kitakobayashi, Mibu, Shimotsuga, Tochigi 321-0293, Japan e-mail: keisuke@dokkyomed.ac.jp

The winter wheat variety Kitahonami shows a superior flour yield in comparison to other Japanese soft wheat varieties. To map the quantitative trait loci (QTL) associated with this trait, association mapping was performed using a panel of lines from Kitahonami's pedigree, along with leading Japanese varieties and advanced breeding lines. Using a mixed linear model corrected for kernel types and familial relatedness, 62 marker-trait associations for flour yield were identified and classified into 21 QTLs. In eighteen of these, Kitahonami alleles showed positive effects. Pedigree analysis demonstrated that a continuous pyramiding of QTLs had occurred throughout the breeding history of Kitahonami. Linkage analyses using three sets of doubled haploid populations from crosses in which Kitahonami was used as a parent were performed, leading to the validation of five of the eight QTLs tested. Among these, QTLs on chromosomes 3B and 7A showed highly significant and consistent effects across the three populations. This study shows that pedigree-based association mapping using breeding materials can be a useful method for QTL identification at the early stages of breeding programs.

To date, few mutations in wheat have been characterized at the molecular level. In this study, the mutations in the three waxy alleles in waxy wheat (Wx-A1h, Wx-B1b and Wx-D1b) were characterized, and waxy gene expression was compared in several wheat lines, including hexaploid and tetraploid waxy lines of wheat. Southern analysis showed that the Wx-B1b allele had sustained a deletion which included the entire coding region of the Wx-B1 gene. DNA homologous to waxy gene sequences was still present in the Wx-A1b and Wx-D1b alleles of the hexaploid waxy mutant. Transcripts of waxy alleles were also detected in both hexaploid and tetraploid mutants at 10 days post-anthesis, but the transcript level was dramatically reduced compared to that found in non-waxy lines. Isolation of cDNAs showed that transcripts were produced by both the Wx-A1b and Wx-D1b alleles. A 23-bp deletion sustained by the Wx-A1b allele at an exon-intron junction results in the use of a cryptic splice site during mRNA processing. The deduced translation product encoded by the Wx-A1b cDNA lacks 39 amino acids, including the putative ADP-glucose binding site and a portion of the transit peptide. The C-terminal region of the deduced protein encoded by the Wx-D1b cDNA lacks the last 30 amino acids. Comparison of the genomic sequences of the null and wild-type Wx-D1 alleles indicated that 588 bp were deleted in the Wx-D1b mutation, and that the last 261 bp of the Wx-D1b cDNA originated from the normally non-transcribed 3' flanking region. Like several deletion mutations characterized in other plant species, both Wx-A1b and Wx-D1b alleles contain small DNA insertions, or filler DNA, between the deletion end-points.

A 79-year-old woman who had diabetes mellitus for 22 years developed coma 1 day after influenza A virus infection. Diabetic ketoacidosis (DKA) was diagnosed based on high serum glucose levels (902 mg/dL), an arterial pH of 7.19 and large urine ketone (3 + ). Subsequently, the patient was treated with insulin and fluid therapy. After improvement in serum glucose levels, diffusion-weighted magnetic resonance imaging (MRI) showed acute infarction in the bilateral middle and posterior cerebral artery territories (Fig. 1). No severe stenosis in major cerebral arteries on magnetic resonance angiography was detected. Transthoracic and transesophageal echocardiography did not show any potential cardiac source of embolism including patent foramen ovale or thrombi in the left ventricle. In the present patient, brain lesions on MRI were located in the white matter, but not in cortices, suggesting microcirculatory impairment by hyperglycemia-induced hypercoagulability and/or hyperviscosity. DKA is associated with thrombotic risks, such as abnormalities in coagulation factors, platelet activation and blood volume, and cerebral edema as a result of DKA might predispose to stroke; however, DKA associated with stroke is rare in adults.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) elicits selective destruction of nigrostriatal dopamine neurons in human and non-human primates along with clinical symptoms of Parkinson’s disease (Langston et al., 1983; Burns et al., 1983). Systemically administered MPTP is biotransformed into 1-methyl-4-phenylpyridium ion (MPP+), which then enters dopaminergic neurons via the dopamine uptake system to destroy nigral cells.

BackgroundEST-PCR markers normally represent specific products from target genes, and are therefore effective tools for genetic analysis. However, because wheat is an allohexaploid plant, PCR products derived from homoeologous genes are often simultaneously amplified. Such products may be easier to differentiate if they include intron sequences, which are more polymorphic than exon sequences. However, genomic sequence data for wheat are limited; therefore it is difficult to predict the location of introns. By using the similarities in gene structures between rice and wheat, we developed a system called PLUG (PCR-based Landmark Unique Gene) to design primers so that PCR products include intron sequences. We then investigated whether products amplified using such primers could serve as markers able to distinguish multiple products derived from homoeologous genes.ResultsThe PLUG system consists of the following steps: (1) Single-copy rice genes (Landmark Unique Gene loci; LUGs) exhibiting high degrees of homology to wheat UniGene sequences are extracted; (2) Alignment analysis is carried out using the LUGs and wheat UniGene sequences to predict exon-exon junctions, and LUGs which can be used to design wheat primers flanking introns (TaEST-LUGs) are extracted; and (3) Primers are designed in an interactive manner. From a total of 4,312 TaEST-LUGs, 24 loci were randomly selected and used to design primers. With all of these primer sets, we obtained specific, intron-containing products from the target genes. These markers were assigned to chromosomes using wheat nullisomic-tetrasomic lines. By PCR-RFLP analysis using agarose gel electrophoresis, 19 of the 24 markers were located on at least one chromosome.ConclusionIn the development of wheat EST-PCR markers capable of efficiently sorting products derived from homoeologous genes, it is important to design primers able to amplify products that include intron sequences with insertion/deletion polymorphisms. Using the PLUG system, wheat EST sequences that can be used for marker development are selected based on comparative genomics with rice, and then primer sets flanking intron sequences are prepared in an interactive, semi-automatic manner. Hence, the PLUG system is an effective tool for large-scale marker development.

Systemically administered 1-methyl-4-phenyl-l,2,3,6-tetrahydropyridine (MPTP), a toxin inducing parkinsonism, is biotransformed into 1methyl-4-.phenylpyridinium ion (MPP+), which then enters dopaminergic neurons via the dopamine uptake system to destroy nigral cells.

Reduction of ethyl 2-acetoxy-3-oxobutanoate (4a) with immobilized baker’s yeast in calcium alginate gel gave a 18 : 19 : 63 mixture of ethyl (2R,3S)- and (2S,3S)-2-acetoxy-3-hydroxybutanoates and ethyl (2S,3S)-2,3-dihydroxybutanoate each with >95% e.e. in 58% yield. A similar treatment of a 2-benzoyloxy analog of 4a afforded a 6 : 94 mixture of ethyl (2R,3S)- and (2S,3S)-2-benzoyloxy-3-hydroxybutanoates (>95% e.e.) in 70% yield. Their absolute configurations were determined by comparison with authentic (2R,3S)- and (2S,3S)-2,3-dihydroxybutanoic acids. Effects of the immobilization and the pH of culture solution on the product ratio are also discussed.

The clinical features of 3 patients from a kindred with adrenoleukodystrophy and the analysis of their plasma sphingomyelin are described. Onset of symptoms was between the ages of 33 and 54 years. Ataxic gait and spasticity were the only symptoms noted during the early stage of the disorder. Dementia and optic atrophy were present in two of the cases. Baseline plasma cortisol was normal, but adrenocorticotropic hormone was elevated. Analysis of plasma sphingomyelin demonstrated an increase in very-long-chain (C24-C26) fatty acid. This study demonstrates that adrenoleukodystrophy may present with spinocerebellar symptoms.

The structure and mRNA accumulation of T-DNA of Ri (A4) plasmids integrated into the genome of Nicotiana tabacum var. Samsun were investigated. Two inserts of TL-DNA were found in the transformed plant (R1) and one (R2) of the progenies obtained by self-crossing of the R1 plant. The expression level of T-DNA-derived transgenic plants was high in roots and stems, but very low in leaves. The gene encoding the major transcript of integrated Ri TL-DNA coincided with the ORF12 (Slightom et al., J. Biol. Chem., 261 (1986) 108).

Systemically administered 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is biotransformed into 1-methyl-4-phenylpyridinium ion (MPP + ), which enters dopaminergic neurons via the dopamine uptake system to destroy nigral cells. Either MPP + is retrogradely transported to the cell body after being taken up at the nerve terminals, or the dopamine uptake sites on the cell body and its dendritic processes are responsible for the toxin directly entering the neuron. Using a 200 μl osmotic minipump, we administered 4 mg of MPTP HCl directly into the unilateral caudate nucleus, i.e., the dopamine nerve terminal area, of monkeys for 14 days. Persistent hemiparkinsonism began to appear in a week. Each monkey exhibited a flexed posture and hypokinesia of the contralateral limbs and circling toward the MPTP-treated side. These disturbances developed within 3 months and maintained a plateau for 3 months until the day of sacrifice. After treatment with apomorphine, there appeared a striking circling away from the MPTP-treated side. Selective cell loss in the MPTP-treated side of the substantia nigra pars compacta was found along the entire rostrocaudal extent relative to the untreated side. In conclusion, MPP + uptake only at the dopamine nerve terminals and retrograde axonal transport to the cell body seemed suffcient to destroy nigral dopamine cells in the monkey.

Reduction of ethyl 2-acetoxy-3-oxobutanoate (4a) with immobilized baker’s yeast in calcium alginate gel gave a 18 : 19 : 63 mixture of ethyl (2R,3S)- and (2S,3S)-2-acetoxy-3-hydroxybutanoates and ethyl (2S,3S)-2,3-dihydroxybutanoate each with >95% e.e. in 58% yield. A similar treatment of a 2-benzoyloxy analog of 4a afforded a 6 : 94 mixture of ethyl (2R,3S)- and (2S,3S)-2-benzoyloxy-3-hydroxybutanoates (>95% e.e.) in 70% yield. Their absolute configurations were determined by comparison with authentic (2R,3S)- and (2S,3S)-2,3-dihydroxybutanoic acids. Effects of the immobilization and the pH of culture solution on the product ratio are also discussed.